yearbook 2013

Transcription

yearbook 2013

ENGLISH
CENTER FOR GENETIC ENGINEERING
YEARBOOK
AND BIOTECHNOLOGY 2013
GENERAL
DIRECTOR’S WORDS
DIRECTIONS
MISSION
PUBLICATIONS
CENTRO DE INGENIERÍA GENÉTICA
Y BIOTECNOLOGÍA
Center for Genetic Engineering and Biotechnology
YEARBOOK 2013
SUMMARY
DIRECTIONS
DIRECTION OF BIOMEDICAL RESEARCH 06
INTRODUCTION
03
MISSION
05
PUBLICATIONS
105
DIRECTION OF TECHNOLOGICAL DEVELOPMENT 44
DIRECTION OF PRODUCTION 47
BIOTHERIUM 52
DIRECTION OF AGRICULTURAL RESEARCH 55
DEPARTMENT OF INFORMATICS AND COMMUNICATIONS 63
DIRECTION OF QUALITY CONTROL 67
DIRECTION OF QUALITY ASSURANCE 71
DIRECTION OF HUMAN RESOURCES 74
HEBER BIOTEC S.A. 77
INTELLECTUAL PROPERTY DIRECTION 82
BUSINESS DEVELOPMENT GROUP 84
DIRECTION OF CLINICAL RESEARCH 88
DIRECTION OF SAFETY AND PROTECTION 94
DEPARTMENT OF REGULATORY ISSUES AND CLINICAL TRIALS 95
CIGB IN Camagüey 99
CIGB IN Sancti Spíritus 102
Project coordinator: Raúl Alejandro Valdés Pavón
Editing and correction: Marta Elizabet Ferrer Cutié, Ernesto Galbán Rodríguez
Compiled by: Berna Eva Molinet, Nayade Beltrán Matos
Design: Raúl Rojas Pérez
Photos: Raúl Rojas Pérez, Jesús Seoane Herrera
© Consejo de Dirección CIGB, 2014
© Elfos Scientiae, 2014
YEARBOOK 2013
General Director’s words
Dear Readers:
The scientific development in Cuba today is the realization of the will, the efforts and
the dedication of all our people to make reality the fact of turning Cuban Science into a
major productive force. Cuban biotechnology as an expression of this effort consists of the
existence of dozens of research-production centers, based on the principle of integration
and closed cycle.
The Center for Genetic Engineering and Biotechnology of Havana (CIGB) is the leading
scientific institution of Cuban biotechnology, with over 1 300 workers, aimed at the obtainment
of recombinant proteins and other novel products, its technological development, production
and commercialization. The positive impact on health, agriculture and the environment is
the key issue of our work. Among our most traditional products already made and marketed,
are: recombinant vaccine against Hepatitis B (Heberbiovac-HB®), recombinant interferon
(alpha and gamma Heberon®), recombinant streptokinase (Heberkinase®), Transfer Factor
(Hebertrans), vaccine against cattle tick (Gavac®) and others. Innovative products have
also been developed, such as QuimiHib® vaccine against Haemophilus influenzae type b
obtained through chemical synthesis, combined tetravalent (Trivac HB®) and pentavalent
(Heberpenta®) vaccines, a bio-product against nematodes (HeberNem®), and a nonhormonall growth enhancer (Aquabio 1) for raising shrimp and fish. All these products used
in Cuba and sold to over 40 countries. In 2006 we also succeeded in obtaining a new
drug for human use unique in the world, named Heberprot-P®, and its sanitary registration
in Cuba (2006), Algeria (2008) and Argentina (2009). It was approved in Venezuela as a
medical service in 2008, and used in the healing of deep and complex ulcers resistant to all
types of conventional treatment in people with the so-called diabetic foot. Its administration
in severely affected patients allowed reducing the risk of amputation by 80 percent. Over
2000 patients were treated in 2009 with very positive results, as part of agreements between
Cuba and Venezuela.
GENERAL DIRECTOR
Dr. Luis Herrera Martínez
Phone: (537) 273 1410
Fax: (537) 273 6008
[email protected]
YEARBOOK 2013
General Director’s words
The development of transgenic maize resistant to fall
armyworm and to glufosinate ammonium has been our main
project in the introduction of genetically-modified plants. It
has demonstrated the high efficacy and safety of this maize.
We work to achieve regulatory approvals in order to be able
to reach a commercializing level in 2009.
A staff of excellence supports the findings and reliability of
CIGB, guaranteed by its scientific-technical and productive
training which is based on the Good Manufacturing Practices
and the existing regulations defining the obtainment of high
quality and safety products.
A summary of the characteristics of the main areas of our
institution, the main lines of research, development, and
products that now characterize the Centre for Genetic
Engineering and Biotechnology are present in this yearbook
Thank you very much.
SUMMARY
YEARBOOK 2013
MISSION
T
he Center for Genetic Engineering and Biotechnology
has an integrative role in the Cuban Biotechnology field,
with high technical and scientific capabilities. It assumes the
responsibility of directly contributing to the socio-economic
development of our country.
Through research it generates knowledge for the development
of new products, services and marketing, based on a quality
system that ensures the satisfaction of our customers, taking
into account the environmental dimension.
Its impact is appointed to human health, agricultural and
aquaculture productions, industry and the environment.
SUMMARY
YEARBOOK 2013
Direction of Biomedical Research
T
he Direction of Biomedical Research (IBM) at the Center for Genetic Engineering
and Biotechnology (CIGB) works in more than 20 projects aimed at the obtainment
and development of biomedical products for the prevention and treatment of infectious
diseases, cardiovascular, neurodegenerative, cancer, inflammation, autoimmunity,
healing and cytoprotection. It comprises the departments of Vaccines, Pharmaceuticals,
Immunodiagnostics and Genomics, Chemical Physics, and Control of Scientific-Technical
Activity and Administrative It counts with 200 workers: 24 % of the researchers obtained the
PhD’s degree in Sciences and 20 % of the staff got the Master’s degree in Sciences.
DIRECTOR
Dr. Gerardo Guillén Nieto
[email protected]
Phone: (537) 271 6022, ext. 7204
Main Executives
Assistant Manager
Head of the Department of Immunodiagnostics and genomics
Dr. Lidia Inés Novoa
[email protected]
Phone (537) 208 4035 (Center of Biological Research)
Department of Vaccines
Head of the Department
Dr. Santiago Dueñas-Carrera
[email protected]
Phone: (537) 271 6022, ext. 7265
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YEARBOOK 2013
The Department of Vaccines is in charge of the obtainment
of prophylactic and therapeutic vaccine formulations against
bacterial and viral diseases by techniques of genetic
engineering. In addition, it develops combined vaccines.
Ongoing Biomedical Projects:
• Therapeutic vaccine against hepatitis B
• Therapeutic vaccine against hepatitis C
• Recombinant vaccine against dengue virus
• Anti-HIV therapy
• Accelular Vaccine against Pertussis
Therapeutic Vaccine against hepatitis B
Leader of the project
Dr. Julio César Aguilar
[email protected]
Phone: (53-7) 250 4541
This Project is in charge of the development of the therapeutic
vaccine Nasvac®, against chronic Hep B virus (HBV) and
works in two immunization strategies with these therapeutic
vaccine candidates by nasal and subcutaneous routes.
The breakdown of tolerance in a transgenic mouse model
had been previously demonstrated constitutively expressing
HBsAg. In 2009, based on histopathological studies in
tg mice immunized with the formulation Nasterap, it was
found that the breakdown of tolerance in the compartments
of humoral and T CD8 + response against HBsAg is not
associated with histopathological damage.
This result, along with others under development in the
project since 1998, enabled the transition to the evaluation
of the therapeutic vaccine candidate against a backdrop
of chronically ill patients with HBV in Dhaka, Bangladesh,
in collaboration with the Liver Foundation of Bangladesh
and along with Japanese collaborators, responsible for the
immunological evaluation of the product.
Clinical Trial (CT) in chronic hepatitis B patients allowed
evidencing the safety of the candidate vaccine when
administered via IN and in a first cycle of five immunizations
and a second cycle of five additional immunizations when
administered IN and SC routes. Additionally an immune
response cytokines in serum of patients vaccinated in
the supernatant of peripheral blood mononuclear cells
stimulated with the antigens of the formulation (HBsAg and
HBcAg) was detected.
Associated to the immune response, a reduction of viral DNA
load non-detectable in 50 % of treated patients could be steadily
seen. Thus, the objectives of the CT phase I-II were completed.
Phase III study in its follow up concluded in 2013. The
current analysis and final report should be concluded during
the first trimester. Results from a Phase III clinical trial have
evidenced the same as those preliminary described from a
Phase I/II clinical trial. This study has been audited by the
national regulatory center and European experts; stating
the satisfactory work in compliance with Good Laboratory
Practices. Greater sustained results of the immune response
related to viral DNA load and the HBsAg seroconversion are
evidenced in the group immunized by the vaccine Nasvac®
compared to the group treated with Peg-interferon.
Vaccine against hepatitis C
Dr. Santiago Dueñas-Carrera
[email protected]
Phone: (53-7) 250-4550
According to recent statistics from the World Health
Organization(WHO), at least 185 million people are infected
with hepatitis C virus (HCV). This is chronic in more than
80 % of affected individuals that often causes cirrhosis and
cancer of liver. There is currently no prophylactic vaccine
available against this pathogen, and only a few candidate
vaccines have reached the clinical phase. The antiviral
treatments in use (of interferon plus ribavirin with the recent
incorporation of Telaprevir or Boceprevir as third element)
are aggressive, expensive and generally not effective in less
than 100 % of cases.
At the CIGB, the structural region of genotype 1b of the
main circulating HCV in Cuba was cloned and is the basis
for vaccine candidates under development. The most
advanced vaccine approach of the Project is a DNA vaccine
formulation based on a mixture of a plasmid containing
genes of the three structural antigens of the virus with
HCV core protein as molecular adjuvant. This DNA vaccine
candidate, called CIGB-250, induces specific humoral
and cellular responses, strong and sustained in different
animal models, with remarkable protection in mice against
challenge with recombinant vaccinia virus expressing the
core gene, E1 and E2, as evidence of immune functional
response. The preclinical phase with this vaccine formulation
was successfully completed, demonstrating its low toxicity in
rodents and stability of its components.
Currently, based on preliminary evidence of immunogenicity
and low reactogenicity observed in individuals immunized
with CIGB 250, a Phase I and II clinical study with a
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YEARBOOK 2013
preventive scenario is also in course in which this candidate vaccine was
administered to individuals with chronic renal failure at risk of acquiring
HCV. The objective of the study is to evaluate safety and immunogenicity
of the vaccine candidate in these patients by three levels of dose, as
well as the ability to avoid HCV after patients require haemodialysis. The
stage with the treatment concluded satisfactorily and the conclusion of
this study is foreseen for the second trimester in 2014.
Another vaccine candidate based on the mixture of HCV structural
antigens: core, E1, E2 and NS3, mixed in a specific proportion formulated
in Aluminum oxide (MixprotHC), has demonstrated its efficacy in animal
models. It is based on inducing humoral and cellular response against
VHC including epitopes stored in viral isolations and taking into account
the immunogenic hierarchy between viral proteins. Immunized mice
developed cross- neutralizing responses of CD4+ and CD8+ T cell
proliferation as well as IFN-γ secretor against Core, E1, E2 and NS3.
In addition, vaccinated mice with MixprotHC showed a marked reduction
of viral titer after they had been elicited with recombinant vaccinia virus
for the structural region of the HCV compared to animals with the wild
virus (WR) and the immunized group with Aluminum oxide (Figure 1).
Figure 1
Log viral title in ovaries
8
6
p= 0.0028
**
4
2
0
Alum
(WR)
Alum
(vvRE)
Mixprot
(WR)
Mixprot
(vvRE)
Groups
Control of the vaccinia virus of hepatitis C (VHC) in BALB/c mice after the challenge with the viaccinia virus recombinant
(vvRE), or wild vaccinia virus (WR). Four groups of (n = 5) Balb/c mice were immunized by intramuscular route, with
MixprotHC (mixture of HCV structural antigens: core, E1, E2 and NS3, mixed in a specific proportion formulated in Aluminum
oxide) or Alum at weeks 0, 2, 4, 8 and 12 the challenge was administered by intraperitoneal route at week 14 with 106 units
forming (uff) of vvRE or WR. The ovaries were collected five days after the challenge. The figure shows a 10- log fold
average viral title ** High significant differences comparing the other three groups and mice challenged with WR; ** p < 0.01,
Kruskal-Wallis Dunn's multiple comparison tests.
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The immunogenicity of MixprotHC was also evaluated in green African
monkeys and its capacity to induce cross-neutralizing antibodies was
evaluated by means of the Neutralization assays VHCcc (Figure 2). At the
same time, T-cell proliferative and IFN-γ secretion response against core,
E1, E2 and NS3 in monkeys after five immunizations with MixprotHC was
detected.
Figure 2
% of infectivity
150
100
50
0
MixprotHC
Groups
Alum
Ab-
The neutralizing activity of purified IgG antibodies in green African monkeys vaccinated with serum MixprotHC( mixture of HCV
structural antigens: core, E1, E2 and NS3, mixed in a specific proportion formulated in Aluminum oxide,). The IgG antibodies
were purified separately and evaluated by means of the Neutralization assays VHCcc (chimeric HCV genotype 1a-2a). Samples
without antibodies were used as negative controls (Ab-). The results were considered positive when infectivity was minor or
equal to 50 % (discontinuous line). These were expressed in percentage of Focus-forming units (FFU) by the VHCcc (FFU in the
presence of antibodies/FFU in the absence of antibodies x 100).
After showing the efficacy of MixprotHC in animal models, the development
of this vaccine candidate will be achieved with clinical trials in 2014.
VACCINE AGAINST DENGUE VIRUS
Dr. Gerardo Guillén
[email protected]
phone: (537) 271 6022, ext. 7204
Every year, 50 to 100 million people in the world suffer from dengue
fever, and 250-500 thousand from dengue hemorrhagic fever. Cuba has
reported five epidemics, and there is constant risk of development of new
events while the vector has not been fully eliminated. Currently, there is
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YEARBOOK 2013
no antiviral drug or preventive vaccine against this disease. The objective
of this project is to obtain a preventive vaccine against all four dengue
virus serotypes (DEN-1, DEN-2, DEN-3 DEN-4), the causative agent of
the disease. The recombinant vaccine project runs jointly by the Tropical
Medicine Institute Pedro Kourí and the CIGB.
Two vaccine strategies are undertaken in the Project: the obtainment of a
vaccine exclusively based on recombinant proteins and the prime-boost
strategy combining recombinant proteins and live attenuated viruses.
Recombinant fusion proteins P64k-DomIII (Neisseria meningitidis protein
fused to domain III of envelope protein of the virus DEN-1 to DEN-4) and
those based on the dengue virus capsid-2 are among the most advanced.
These constructions have been obtained in Escherichia coli with high
level of purity (Marcos E et al., Arch Virol. 2013;158:225-30). Besides,
They have been evaluated in mice, with evidence of functional response
against infectious virus, (Figure 3) (Valdés I et al., Virology. 2009;394:24958; Izquierdo A et al., Antivir Res. 2012;95(1):1-8).Additionally, after
having established the animal in non primates (macacus, green monkey)
models for dengue infection of the virus DEN, (Martín J et al., Microbiol
Inmunol. 2009;53:216-23; Valdés I et al., Int J Infect Dis. 2013) protection
and prime-boost effect in combination with infectious virus was evidenced.
infectivo (Valdés I et al., Int J Infect Dis. 2010;14:e377–e383; Valdés I et
al., Clin Vaccine Immunol. 2011;18(3):455-9). In the preclinical phase of
evaluation, the monovalent vaccine candidate based on virus capsid D2,
administered to monkeys, provided protection against the viral infection.
The tetravalent formulation gave protection against the four viruses in
mice. Its evaluation in monkeys is planned for 2014.
Figure 3
Percentage of survival
100
DENV-2
75
Aggregate DIIIC-2
50
*
*
25
0
*
*
*
*
*
*
*
DIIIC-2
Placebo
0
3
6
9
12
15
18
21
Days after the viral challenge
It represents the survival curves of immunized mice with the recombinant protein DIIIC based on capsids of the s (DENV), and
challenged intracranially with a lethal host of combining DENV-2 with DIIIC.
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Vaccine against the human immunodeficiency virus
and AIDS
Dr. Enrique Iglesias
[email protected]
Phone: (53-7) 250 4541
Since it was identified in 1981, acquired immunodeficiency syndrome
(AIDS) has killed more than 25 million people. It is one of the most
devastating epidemics in human history. The organization responsible for
coordinating efforts in the struggle against AIDS (UNAIDS) estimates that
worldwide prevalence in recent years has been increased, but there is a
speed in the steady increase of infected people. By the end of 2012, 32.238.8 million people lived with the immunodeficiency virus type-I (HIV-1).
That same year, the number of cases increased by 1.9 and 2.7 million
approximately; and 1.4 and 1.9 million died with HIV positive (adults and
children). In the area of access to anti-retroviral terapy (ART) reached
to 64 % to people in need in poor countries (according to the guide
of treatment of the World Organization of the Health (WHO in 2010).
However, after updating the therapeutic guide in 2013, only 34 % of the
eligible ones were treated. On the other hand, in some countries of the
sub-Saharan Africa, the retention levels of the ART are below 50 % two
years after initiated. Hence, the financing to the therapies must increase
and it is necessary to go foreseeing the need to go on from the generic
compounds, at present in use, to treatments of the second and third
line in the next years due to the dissemination of resistant isolations. In
this context, researchers could probably develop an effective or partially
effective vaccine to offer protection to seronegatives, and to replace
whole or partially the existing therapies in seropositive ones, which do
not eliminate the infection and undesirable and incompatible side effects
in patients will appeared in long terms.
This project is aimed at developing a vaccine candidate against human
immunodeficiency virus type 1 (HIV-1). Based on this goal, a multiantigenic
formulation vaccine candidate TERAVAC was obtained which comprises
the recombinant multiepitopic protein CR3 that contains T cell epitopes
and the surface and nucleocapsid antigens of the Hepatitis B Virus
(HBV) to induce Th1 cellular immune responses in both the systemic and
gastrointestinal tract compartments.
CR3 recombinant protein does not exist in nature in any host but is made
up of different fragments of proteins present in HIV-1 like: the envelope
glycoprotein (gp 160), vpr protein (p15), pol (p66), nef (p27) and gag
(p24). These regions are rich in antigenic epitopes for T cell helper and
cytotoxic type also found for B-cell epitopes. This ensures a high number
of epitopes restricted by a wide range of HLA haplotypes I and II. However,
according to studies conducted in mice, CR3 protein, by itself, is not able
to induce a Th1 response or stimulate CD8+ T lymphocytes. Therefore,
when the CR3 is immunized along with structural antigens of hepatitis B
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YEARBOOK 2013
virus, humoral and cellular Th1-type are obtained in the gastrointestinal
mucosal compartment (Figure 4) and the systemic with stimulation of antiHIV CD8 T lymphocytes (Figure 5). In addition, the studies have shown
that TERAVAC could generate a Th1 type response in the presence of an
ongoing immunization-driven a Th2 type response. This result suggests
Figure 4
A
B
CR3
RPMI
Con A
Placebo (sc+in)
C+S (sc+in)
CR3+C+S (sc+in)
Detection of IFN-γ secretive cells in mice mesenteric lymph nodes against
the protein CR3A) Ten days after the last administration, immunocytes cells
from jejunal and colonic lymph nodes were extracted (A). ). Then, these
were stimulated with protein CR3 and controls. Finally, ELISPOT plates
were processed and results representative of duplicate wells (B). CR3,
are sampled, cells stimulated with protein CR3; RPMI, cells incubated with
complete medium, Con A, cells stimulated with Concanavalin A., (Con A).
i.n.: intranasal route s.c.: route subcutaneous. C: HBcAg. S: HBsAg.
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Figure 5
CD8APC
Not stimulated
Con A
CR3
1000
1000
1000
100
100
100
10
10
10
1
1
1
0.1
0.1
1
10
100
1000
0.1
0.1
C+S (sc+in)
1
10
100
1000
0.1
0.1
C+S (sc+in)
1000
1000
1000
100
100
100
10
10
10
1
1
1
0.1
0.1
1
10
100
CR3+C+S (sc+in)
1000
0.1
0.1
1
10
1
10
100
1000
10
100
1000
C+S (sc+in)
100
CR3+C+S (sc+in)
1000
0.1
0.1
1
CR3+C+S (sc+in)
Proliferative response of T CD8+ lymphocytes. Homogeneous mixtures of spleen cells from mice were stained with CFSE
and cultured for five days with culture medium, Concanavalin A( with A) and CR3. Ten thousand viable lymphocytes and blasts
(R1) were selected for the analyses. The frequency of T CD8+ CR3- cells specific that were divided in each group after ex
vivo stimulation was determined subtracting the percentage of cells CD8+ CFSElow (Q1) taken from the culture not stimulated to the percentage of CD8+ CFSElow of the stimulated splenocytes with the antigen CR3. i.n.: route intranasal. s.c.: route
subcutaneous. C: HBcAg. S: HBsAg.
a therapeutic benefit to restore anti HIV cellular response in
seropositive patients. Finally, induction of anti HIV cellular
and humoral responses has been demonstrated thus; this
vaccine candidate could be considered bivalent: anti-HIV
and anti –VHB. During 2009 and 2010, progress was made
in preclinical safety testing, local toxicology and others, and
the results were satisfactory. Then, the file to request a Phase
1 clinical trial with this vaccine candidate was arranged by
the State Center for Drug Control (Cecmed).The required
authorization was given in 2010. During 2011-2013, the
immunization program in the first group of 9 patients and the
recruitment has been completed.
Project Accellular Pertussis Vaccine
MSc. Anabel Álvarez
[email protected]
phone: (537) 2716022 ext. 7261
Epidemiology of the infection by Bordetella pertussis.
Whooping cough which is caused by the bacterium Bordetella
pertussis, is currently the most predominant vaccinepreventable disease around the world. The frequency of
infection calculated by antibody titers against pertussis toxin
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YEARBOOK 2013
was reported recently with values between 1 - 6 % (Cherry, JD.
Clin Infect Dis. 2010;51(6):663-7).
The essential characteristics are:
• Cyclic outbreaks every 2 or 5 years where population’s
immunization coverage is > 80 - 90 %.
• High frequency of cases in children aged under 1 and
10-year-old children.
• Increase of the infection rate in teenagers between
11-18 years old and adults.
The main cause associated with the epidemiological behavior
of B. pertussis is the fact that neither the vaccines nor the
natural infection confer lifetime protection comparing to
other diseases. Therefore, the microorganism circulates and
evolves in the immunized populations. Duration of immunity
from vaccines is extended to 4-12 years and the natural
infection between 4-20 years (Wendelboe AM. Pediatr Infect
Dis J. 2005;24(5):S58).
In the last 10 years, the number of publications reporting an
increase of the pertussis incidence is notorious. Among the
possible causes trying to explain the new outbreaks and the
increase are:
• Major conscience about the disease for the medical staff
who report an increasing number of cases. Additionally,
the introduction of polymerase chain reaction testing
of genetic material (known as DNA PCR) and its wide
use in developed country have contributed to the
number of reported cases (Cherry JD, N Engl J Med.
2012;367(9):785-7).
• WHO has stated that the best acellular vaccines (with
at least 3 components) can be less effective (85-88 %)
than the best cellular vaccines, (96 %) (WHO Weekl
Epidemiol Record. 2010;85(40):385-400).
• The evolution of the microorganism towards variants
more virulent (Advani et al. Vaccine. 2011;29(18):343842; Octavia et al. J Infect Dis. 2012;205(8):1220-4).
A very important aspect is related to the incidence studies
which suggest that the current immunization program have
to be re-evaluated, since they are ineffective to break the
increase of the cases and to eradicate the disease. It is
demonstrated that the immunization programs must begin
closer to the birth (0-1 month). The Centers for Disease
Control (CDC) recommend to implement booster dose
to pregnant mothers (20 weeks) and practice the cocoon
strategy (Gall SA. Clin Obstet Gynecol. 2012;55(2):498509. In the last 5 years, the number of infected cases has
increased in Europe (Sweden, Holland) and Australia,
Bordetella pertussis strains with the pertussis toxin
promoter allele ptxP3 and the pertactin protein type 2
(Prn2 (King, et al. BMC Genomics 2008;9:311; Advani et al.
J Microbiol Methods. 2009;78(3):297-301; Advani et al.
Vaccine 2011;29(18):3438-42; van Gent et al. PLoS
ONE. 2011;6(5):e20340; Octavia, et al. J Infect Dis.
2012;205(8):1220-24). In other countries like Japan, the
number of strains that do not express the protein Prn1 has
increased (Otsuka et al. PLoS ONE. 2012;7(2):e31985).
Studies suggest that strains ptxP3, Prn2 are more virulent
because they express high levels of pertussis toxin
and escape more easily to the immunity of the current
vaccines (contain antigen Prn1) during the declination of
the immunity. This behavior of the microorganism evolution
suggests that the current vaccines must be updated and
technologically improved.
This mutation could inactive 100 % the toxin pertussis
without affecting immunological properties and adjuvants
of the protein The project strategy is based on the
development of an acellular vaccine containing three
components PTg, FHA and Prn2. Particularly, genes for
pertactin and toxin pertussis present in vaccine strains are
ptxA2 and Prn2, which have a major appearance frequency
in the circulating strains. Moreover, vaccine strains
containing mutated gene ptxA2 (9K/129G) genetically
affecting the ADP-ribosyltransferase activity of toxins. This
mutation inactivates 100 % pertussis toxin without affecting
the immunological and adjuvant properties of the protein
(Del Giudice and Rappuoli).
The project arranges a fermentation process at a scale of
20L where a chemically defined mean is used and high
levels of expression of the three antigens PTg, FHA and Prn
are achieved using a genetically detoxified strain The three
antigens are purified at high levels of purity and low levels of
endotoxin, also the establishment of the essays ELISA are
completed for the detection and/or quantification of the three
antigens which allow to give a follow-up during the process
of obtaining the antigens.
Using the antigens with the developed methodology, the
protection was demonstrated in the mouse's model MICA
(challenge intracranial modified WHO) with an acellular
formulation (in proportion PTg:FHA:Prn2 of 5:2.5:2.5). The
functional activity was highly significant and superior to the
not immunized control group.
During 2013, the project advanced in the conditioning of
the development area at the Carlos J. Finlay Institute to
work with B. pertussis, with the preparation of reference
lyophilized targets and glycerol work. Another point was,
in the establishment of fermentation at 20L scale, the
centrifugation replacement by tangential microfiltration
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YEARBOOK 2013
during the clarification of the fermentative culture, which reduces
the operation times and allows the extraction operations of the
pertactin protein, eliminating in turn the possible biological risk.
In this sense, a procedure was established to work safe and
free of biological risk with culture supernatant in the purification
stages.
Department of Pharmaceuticals
Dr. Marta Ayala
[email protected]
Phone: (537) 250 4494
In 2013, the research, the stock of evidences of mechanism of
action, and the development of new pharmaceutical products have
remained on working for the treatment of cancer, cardiovascular
disease, macular degeneration, and neurodegenerative
diseases, healing, cytoprotection, and autoimmune diseases as
well as the development of therapeutic candidates against the
virus of human immunodeficiency (HIV). The clinical researches
for the assessment of new candidates for the treatment of solid
and hematological tumors to superior stages.
Other therapeutical targets of interest are currently in course
researches.
Ongoing Projects:
• CIGB-300: antitumor peptide which inhibits the CK2mediated phosphorylation;
• CIGB550-E7: therapeutic vaccine: fusion of the HPV E7
antigen to an immunostimulatory peptide;
• CIGB-552: antitumor peptide through inhibition of the
transcription factor NFkB;
• CIGB-247: anti VEGF vaccine;
• CIGB-166: anti-VEGF therapeutic recombinant antibody
fragments identifying VEGF-A through a novel epitope
• Heberprot-P®: Study of differential expression in patients
•
•
•
•
•
•
•
•
treated with Heberprot-P®;
Diabetic Neuropathy: molecules with neuroprotective
capacity;
CIGB-500: peptide with broad cytoprotective capacity;
CIGB-530: peptide with anti-fibrotic capacity;
CIGB-540: peptide for the treatment of hypertrophic wounds;
CIGB-845: therapeutic combined vaccine for neuroprotection;
CIGB-50: therapeutic vaccine to induce anti-IL-15 antibodies;
CIGB-55: IL-15 antagonist peptide;
CIGB-210: peptide with antiviral action.
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YEARBOOK 2013
CIGB-300: antitumoral peptide which
inhibits the CK2-mediated
phosphorylation
Dr. Silvio Perea
[email protected]
Phone: (53-7) 250 4486
The CIGB-300 (peptide which inhibits the CK2-mediated
phosphorylation) focuses on the search for drugs with
therapeutic action in cancer, and the development of
peptide inhibiting intracellular action directed against
mediated phosphorylation by CK2 (casein kinase 2 or CK2).
This enzyme is of the serine-threonine kind and is often
deregulated in many human tumors. The research of our
project has been designed taking as a hypothesis that the
abrogation of phosphorylation of CK2 can induce antitumor
effect in different locations.
The project’s leading product, CIGB-300, induces apoptosis in
different tumor cell lines (human papilloma virus (HPV +); lung;
breast and others). Figure 6 illustrates the results after using
CIGB-300 on the in vitro inhibition of CK2 phosphorylation
and activation of caspase, as well as the anti-tumor effect in
animal models. These results are discussed in detail in an
article published in Cancer Research.
Currently, CIGB-300 peptide is being developed as
the obtainment of a peptide inhibiting CK2mediated
phosphorylation, present in 99.7 % of patients with cervicuterine cancer.
For this purpose, libraries of peptides have been tested in
filamentous phages, in order to find E7 binding ligands whose
sequences are chemically synthesized and the peptides
could be tested in cellular models of transformation by HPV
(CaSki cells) and in animal models with human tumors in
athymic mice. Because E7 oncoprotein is mostly expressed
in the nucleus, the foreseeable use of a system providing
peptides of intracellular action is expected. Different
peptides that enter into the cell have been described and
used for the transport of macromolecules and peptides
into compartments such as: the nucleus, the cytoplasm,
endoplasmic endothelial and the mitochondria.
Figure 6
A
B
Negative control
CIGB-300
Peptide NR
Relative levels of phosphorylation
6000
5000
Tumor volume (mm3)
C
4000
3000
2000
1000
2.0
1.5
1.5
0.5
0
0
2
4
6
8
10 12 14 16
18 20
22
Control
CIGB-300
Tat
Effect of CIGB-300 in tumor cells. A) Antitumoral effect in animal model. Peptide NR: peptide not related. B) Inhibition in vitro of the
phosphorylation of CK2. C) Activation of caspase (apoptosis).
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YEARBOOK 2013
From the mechanistic point of view, it is known that protein B23/
nucleophosmin constitutes one of the substrates of CK2 mostly inhibited
by the action of the CIGB-300. This inhibition leads to the nucleolar
disassembly followed by apoptosis on tumor cells. Interestingly, CIGB300 has demonstrated greater in vitro effectiveness in tumor cell which
apparently because inhibits B23/NPM phosphorylation in different cancer
cell lines.
CIGB-300 finished the study stages of pharmacological, toxicological
concepts and reached the clinical evaluation. In 2006, the first Phase
I clinical trial was achieved with this peptide in patients with high grade
cervical-uterine cancer by intralesional route. The results indicated that
it is tolerant and safe, also evidenced some signs of efficacy. Later,
CIGB-300 has continued its evaluation in patients with cancer by
different clinical studies which allow to know the pharmacokinetic and
biodistribution profiling of the product, also confirming safety y tolerance
after its administration by intravenous route.
CIGB550-E7: vaccine against el virus papiloma
humano
Dr. Isis Torrens
[email protected]
Phone: (537) 250 4470
Cervic-uterine cancer continues being a serious health problem for women
worldwide. Based on reports from the World Health Organization, there
are approximately 500 000 affected women. In some regions of Latin
America, for instance, cervical cancer ranks as the most frequent tumor
in women, followed by breast cancer. In Cuba, despite of the existence
of a national program aimed at early detection; 370 women die every
year because of this disease. The development of this tumor has been
associated with infection with human papillomavirus (HPV), classified
as high-risk; in which HPV-16 and HPV-18 are the most frequent ones.
Based on this background, searching for therapeutic alternatives that may
emerge from scientific knowledge for the treatment of cervical cancer is
a priority in health programs in many countries.
High degree cervical intraepithelial neoplasia (CIN) can be easily and
effectively treated by local excision or destruction. However, morbidity
can be associated with the treatment procedure and recurrence control
of the disease. Moreover, patients on advanced stages of illness yet are
usually immune-competent, so they can receive more benefits from this
therapy. Thus, antitumor therapeutic vaccines could be more effective.
The project, called CIGB550-E7 with its own registry, is focused on
finding therapeutic alternatives for the treatment of cervi-uterine cancer
with a vaccine approach and two forms of vaccine.
Vaccine CIGB550-E7 consists of a covalent bond of E7 protein and
CIGB550 peptide, which is expressed efficiently in Escherichia coli
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YEARBOOK 2013
as a fusion protein and with a high aggregation status. CIGB550 is
recently described by our group as a penetrator peptide of cells, with
immunostimulatory properties. It is fused to the E7 protein of HVP-16,
transports E7 protein to the inner of the cells, favors the generation of a
potent specific cellular immune response, and exert its effect on tumors
that express the Ag E7 of HVP-16.
The therapeutic vaccine CIGB550-E7 completed its proof-of concept test
and it is currently in its development study. This candidate must begin its
clinical development phase in 2015.
Figure 7
A
B
C
D
Evaluation of the penetrating capacity to cells of the protein of merger
CIGB-550-E7 by confocal microscopy. The murine splenocytes incubated
with: A) 1.66 μM of the peptide CIGB-550 biotinylated; B) 1.66 μM of the
protein of merger CIGB-550-E7; C) 1.66 μM of the protein E7; and D) with
phosphate buffer saline (PBS).
New antitumor peptides and immunomodulators
derived from protein Limulus anti-LPS factor
Dr. Maribel Guerra Vallespí
[email protected]
Phone: (537) 250 4491
In spite of the progress of modern medicine, cancer became the
second cause of death in Cuba: 19 thousand people die annually
suffering from cancer and about 29 thousand new cases are diagnosed.
Therapies against important molecular targets for survival, proliferation
and dissemination of the tumor cell, are changing paradigm of cancer
treatment and will probably be used in most of the patients over the next
10 years.
This research and development project is aimed at obtaining new drugs
to treat cancer based on the identification of new antitumor peptides,
derived from a scanning of Ala from the 32-51 region of Limulus protein
anti-LPS factor. The leader product, CIGB-552, is a synthetic peptide
that inhibits intracellular action of the transcription factor NFkB. It is an
important element that activates genes involved in resistance to apoptosis
in cancer cell, stimulates the expression of oncogene, angiogenesis and
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YEARBOOK 2013
cell proliferation. Besides, it protects lab animals of double
challenge (firstly protected animals from the first challenge
with tumor cell receive a second stroke), the effectiveness of
this molecule providing a sustained antitumor effect which
evidences the development of a specific adaptive immune
response and inmunomodulating activity of this new peptide.
Figure 8 shows the cytotoxic effect of CIGB-552 in different
tumor lines, as well as inhibition of the subunit RelA(p65) of
NFkB factor and the induction of apoptosis in the cancer cell
(Figure 8).
Figure 8
A
300
CIGB-552
250
IC50
200
150
100
50
0
H-82
H-125
HT-29
LS174T
MCA-231 MCF-7
PBMC
CIGB-552 (150μM)
B
NT
Sh
12h
24h
ReLA (p65)
NFkB
C
CIGB-552
Cell with
nuclear
fragmentation
(Apoptosis)
Cytotoxic effect of the CIGB-552. A) Cytotoxicity in different tumor lines, in absence of toxicity in normal cells. B) Inhibition of the
subunit RelA (p65) of the factor NF-kB in the tumor line of colon cancer HT-29 treated with CIGB-552. NT: not treated cells. C)
Apoptosis mediated by the CIGB-552 in cells of breast cancer (MCF-7) evaluated by microscopy confocal. PBMC: mononuclear not
tumor cells of peripheral blood.
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YEARBOOK 2013
CIGB-552 has demonstrated efficacy in diverse tumor
models on immune-competent mice: TC-1 model (epithelial
lung cells), myeloma B16, Lewis carcinoma (lung metastases
model), and myeloma X-63, in a colon carcinoma model in
Balb/c mice and in a xenograft model of human colon cancer
HT-29 (Figure 9). The action of this molecule is based on
the combination of a straight cytotoxic effect upon cancer
cell and to provide immune elements leading to revert host
tolerance to tumor.
Figure 9
Average antitumoral volume (cm3)
A
CIGB-552
(0.72 mg/kg) 78.5
600
500
CIGB-552
(1.44 mg/kg) 80.5
400
Saline 82.2
300
200
100
0
0
5
10
15
20
25
30
Postimplant days HT-29
Density of vessels
(% of pixels/field)
B
0.015
Number of microvessels in the tumor
0.010
P < 0.05
P < 0.01
CIGB-552
0.7 mg/kg
CIGB-552
1.4 mg/kg
0.005
0
Saline group
Systemic administration of the CIGB-552 in an implant model of colon tumor of human HT-29. A) antitumoral Efficacy of the
CIGB-552. The arrows indicate the subcutaneous administration of the CIGB-552. B) The CIGB-552 demonstrates a significant antiangiogenic effect in an implant model of human colon tumor, calculating the number of microvessels in the tumor.
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YEARBOOK 2013
The researches of our project are aimed at developing
anticancer molecules taking into hypothesis the discovery
of a new mechanism to regulate anti-apoptotic activity of
the NFkB factor on tumor cell, stimulate host immunity and
control inflammation.
Laboratory of Cancer immunotherapy
Leader of the projects
Jorge V. Gavilondo Cowley
[email protected]
phone: (53-7) 250 4480
The Laboratory of Cancer immunotherapy of the
Pharmaceutical
department
develops
therapeutical
antibodies, mainly against cancer. Some of them can also
be used in the treatment of other chronic human diseases
which pathogenesis or sympthomatology is affected by
the vascular endothelial growth factor (VEGF) and other
proangiogenic molecules. The laboratory develops two
projects: CIGB-247 and CIGB-166a.
CIGB-247: Active Immunotherapy
(therapeutical vaccines) of cancer and
other diseases using antigens related
to tumor stroma
In the last 10 years, the experimental and clinical information
that highlights the special function exerted by stroma of
malignant tumors in their origin, growth and dissemination
has substantially increased. Molecules produced by tumors
and cellular elements of stroma stimulating blood and lymph
vessels are examples of new therapeutical targets against
cancer.
The therapeutic vaccines that are being developed in this
project have been designed to achieve their effect based
on the development of antibodies blocking the pro-tumoral
actions related molecules as antigens. In addition, these
vaccines are used to develop cytotoxic cellular responses
against cells of the tumor stroma or the tumor itself. Moreover,
they can promote other immune responses associated to
epitope spreading due to the use of potent adjuvants in the
vaccine preparations studied.
According to the nature of effector mechanisms, therapeutic
vaccines have potentially less prolonged and undesirable
side effects. These therapeutic vaccines are valued as
additional and complementary treatments to previous stated
ones against cancer like chemotherapy, radiotherapy and
also the conventional anti-angiogenic. According to the
nature of the effective mechanisms, therapeutic vaccines
have potentially fewer undesirable side effects, which it is an
advantage to their administration.
In this project, the main tumor targets to antigens using
recombinant DNA technology are molecules from the family
of vascular endothelium growth factor (VEGF), its receptors,
and other spreading fibroblasts present in tumors.
The first vaccine candidate as antigen proposed for
this project was the human VEGF-121 isoform. Studies
demonstrated that immunization with naked DNA encoding
human VEGF-121 isoform had antitumor effects in the model
C57Bl/6 mice bearing B16-F10 melanoma tumors (BequetRomero et al., Angiogenesis. 2007;10:23-34).
After these first studies, a recombinant antigen representative
of the human VEGF-121 was expressed in Escherichia coli
as a fusion protein with P64K of Neisseria meningitidis (Nm).
Its value as a carrier and immune stimulant of the response
has been documented. This antigen, called P64-VEGFKDR-,
was combined with various adjuvants (alumina, VSSP and
DDA/TDB) administered subcutaneously in schemes of
prophylactic - therapeutic immunization. The vaccination
prolongs the implementation time of B16-F10 melanoma
in C57Bl/6 mice, reduces its growth and increases mice
survival (Morera et al., Angiogenesis. 2008;11:381-93).
These researches demonstrated that the immunization
produced an IgG antibody response against the human
antigen and its murine VEGF. Moreover, the sera of immunized
animals blocked cell proliferation of the human umbilical
cord cells (HuVEC), stimulated in in vitro with human VEGF,
and inhibited the interaction between the human VEGF
and its receptor KDR. In addition, it was demonstrated that
immunized mice developed specific cytotoxic spleen cells
against different tumor cell lines secreting VEGF derived
from C57Bl/6 mice. The treatment of mice with anti-mouse
CD8 monoclonal antibodies reduced the final antitumor effect
of the vaccine; evidencing the possible antitumoral effect of
the vaccine based on humoral and cellular mechanisms
The combination P64-VEGFKDR- and the adjuvant VSSP
were selected to develop the first vaccine candidate, called
CIGB-247V. Additionally, preclinical studies were conducted
using prophylactic-therapeutic immunization schemes in
Balb/c mice bearing other experimental tumors. It was
demonstrated that CIGB-247V vaccine candidate reduces
tumor volume and increases animal survivals in mice bearing
colon tumor CT-26 or F3-II breast cancer. The vaccine also
reduces the number of artificial metastases provoked in lungs
after CT-26 or 3LL-D122 cells are injected by intravenous
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YEARBOOK 2013
route, as well as spontaneous lung metastasis 3LL-D122 and
F3II (Bequet-Romero et al. Vaccine 2012;30(10):1790-1799).
As in the case of the strain C57BL/6, the induction of an
IgG antibody response and its murine counterpart was
demonstrated by this vaccine candidate in Balb/c mice. The
immunized sera inhibited the interaction between human
VEGF and its receptor KDR. Spleen cells of immunized
animals, previously incubated with the antigen, showed
specific cytotoxicity against tumor cell lines from Balb/c
producing VEGF.
The vaccine candidate CIGB-247V has also been studied
in other species. In rats, rabbits and non-human primates,
the vaccine induced an IgG antibody response blocking
antibodies and a cellular response targeting tumor cells
producing VEGF The anti-VEGF antibodies decreased
progressively after finishing the induction phase (8
immunizations in 8 weeks),not avoiding its increase injecting
booster doses in rats, rabbits and non-human primates,
which suggests the existence of immunological memory.
There are no scientific paper publishing hematological
adverse effects, after administering the vaccine to these
species, nor in parameters of blood biochemistry. No
histological affectations have been observed of critical
organs, nor important alterations in the capacity of rats
or non human primates to cure deep skin wounds in skin
(Morera et al. Vaccine. 2010;28:3453-61; Morera et al.
Vaccine. 2012;30:368-77).
The vaccine candidate CIGB-247V complies with the
standards of Good Manufacturing Practices (GMP) were
successfully completed after the preclinical toxicological
studies Therefore, in 2011 this vaccine obtained the
permission by the CECMED a phase I clinical trial in
patients with advanced cancer code name CENTAURO.
In that multicenter, Individuals were subcutaneously
immunized for 8 consecutive weeks, at three antigen dose
levels, and re-immunized on week twelve. The study was
finished in mid-2012, demonstrating that the vaccine was
safe and immunogenic. On week sixteen, evaluations
of safety, tolerance, clinical status, and immunogenicity
(seroconversion for anti-VEGF IgG, serum VEGF/KDR-Fc
blocking ability, and gamma-IFN ELISPOT with blood cells
stimulated in vitro with mutated VEGF) were done. CIGB-247
was immunogenic and higher numbers of individuals positive
to the three immune response tests were seen with increasing
antigen dose.
Surviving patients in the follow-up stage of the clinical trial
CENTAURO has continued for at least two years. Patients
maintain a monthly vaccination. The safety profile keeps
on being excellent. There are two patients with complete
response and some are stable. These results are included
in a recent publication (Gavilondo JV et al. Vaccine. 2014;
in press).
According to the results obtained in the study CENTAURO, in
2013 a new phase Ib clinical trial began called CENTAURO-2,
in which new antigen doses are explored of the adjuvant
VSSP in the vaccine, as well as groups with aluminum
phosphate as adjuvant are included. This trial is currently in
course and must be finished in the middle of 2014.
The vaccine candidate CIGB-247 is potentially applicable to
other diseases associated with excessive and pathological
angiogenesis, due to an increase in VEGF; among them
age-related macular degeneration (AMD) and rheumatoid
arthritis. The therapeutic vaccine candidate CIGB-247 is
protected by a patent approved in a dozen of countries
(CU2002-0076 anti-angiogenic active Immunotherapy).
In this project, other vaccine antigens derived from molecules
of the tumor stroma are also being in research.
CIGB-166a: Passive immunotherapy with
recombinant antibodies specifically
recognizing VEGF
Unlike the above project, the immunotherapeutic strategy of
this project is based on administering recombinant antibodies
to patients with cancer. The recombinant antibodies are
among the biotech molecules’ largest market around the
world. They have a wide spectrum of usage in acute and
chronic.
In this project, we began to provide a technological platform
that allows us to obtain antibodies of human origin, and to
avoid potential rejection reactions in a future therapeutic
application in human beings It is based on the obtainment
of a library of human antibody fragments type Fv of simple
chain (scFv), displayed in filamentous phages, constructed
from human lymphoid cells of different origin (Rojas et al.,
Biochem Biophys Res Commun. 2005;336:1207-1213).
From this library, it is possible to select combination sites
of human antibodies (in scFv fragments) against a lot of
different antigens without the need for immunizations or
lymphoid sampling. As part of this platform, for inducing
therapeutic molecules in this project, mammalian cells and
bacterial vectors have also been developed to produce scFv
antibody fragments or complete antibody molecules with
effector functions, respectively.
Currently, the project is focused on the development of
therapeutic antibodies that block their pro-angiogenic effects
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YEARBOOK 2013
in which pathogenesis or symptomatology are associated
with an excessive VEGF production, such as cancer or AMD.
The most advanced molecules are a scFv fragment called
CIGB-166a, and its bivalent form where two combination
sites as scFv are associated with Fc and human IgG1called
CIGB-598a. The first one is expressed in bacteria, while the
second one is produced by CHO cells. The original binding
site(called 2H1) was selected from a library of phages through
a strategy of selection based on a modified VEGF as antigen,
to increase the possibility to obtain antibodies against new
epitopes of thel VEGF (Lamdan, H et al. Journal Biotechnol.
2011;151(2):166-74). Later, the combination site 2H1 was
modified through specific mutations in the domains CDR3
of both variable chains of the scFv, in order to increase the
affinity of the VEGF (CU 2010-264. recombinant antibodies
against VEGF obtained by mutagenesis of variable regions).
CIGB-166a and CIGB-598a are constructed from this new
combination site, with a greater affinity by the human VEGF
(Lamdan, H. et al. 2013; Molecular Biosystems 9:2097).
These molecules are new because they recognize an epitope
in the human VEGF not previously reported. The interaction
of these molecules with VEGF disables the VEGF ability to
bind VEGFR1 y VEGFR2 receptors. Additionally, both CIGB166a and CIGB-598a recognize some VEGF, neutralize the
proliferative effects and promote the migration of the HuVEC
induced by VEGF affecting the formation of tumor cells.
Currently, the antibody fragment is now in development
and a suitable formulation is under study. In the case of
the bivalent construction, a certified cell bank of producer
transfectoma is built.
In 2013, this Project has opened developing tasks and library
constructions of human nanobodies which are in course.
Healing and citoprotection
Dr. Jorge Berlanga Acosta
[email protected]
Phone: (53-7) 250 4479
Heberprot-P®: Diabetes and Healing
The project of healing and cytoprotection has led to a
comparative study of the differential kinetic expression of
proteins at the granulating tissue of the diabetic foot ulcers,
ischemic, and neuropathic grades III and IV before and after
receiving treatment with Heberprot-P®. It was demonstrated
that the treatment favors the local expression of the
epidermal growth factors (active pharmaceutical ingredient
of Heberprot-P®), protein expression of extracellular matrix
and markers related to the replicating competence of cells
Some of these findings may be hypothetically related with
low recidive index in treated wounds. Currently, these studies
have been extended to fibroblasts in culture obtained from
patients’ ulcers. The expression of genes related with healing
is compared to those expressed in the granulation tissues of
other types of wounds like burns. The impact of simulated
“hyperglycemia” in the culture media on gene expression of
skin fibroblasts is also studied.
Among the most important findings obtained during the
2013 are quoted in the treatment with Heberprot-P® by
correcting enzymes and mediators with REDOX balance
in peripheral blood. Paradoxically, these effects are not
primarily corrected in tissue samples of granulation finishing
the treatment scheme. In 2014, it is expected to continue
and expand these studies to diabetic patients without active
ulcers, metabolically compensated and not compensated.
Likewise, it is expected to demonstrate a hypothetical
interrelation between the levels of glycated hemoglobin
and the proteinases circulating levels. In addition, it was
demonstrated that a number of genes with aberrant
expression in those organs responsible for clearing 70 % of
blood glucose in the involved organs, such as the liver, the
skeletal muscle and the adipose tissue; which have been
considered historically as Type 2 Diabetes Risk Markers are
expressed identically in the cells of granulation tissue of the
lower limb ulcers in our patients. Therefore, the epigenetic
mechanisms involved on gene expression related to glucose
metabolism and the oxidative phosphorylation in diabetes are
validated and confirmed. Finally, our group has contributed
by means of PCR and immunohistochemical studies to the
understanding of molecular bases of the process of wound
chronification in diabetic ulcers, in which ischemia of the
lower limbs is an aggravating factor.
CIGB-500. Cardioprotective agent in
acute myocardial infarction
According to circumstantial and collateral evidences obtained
by our group of a possible trophic effect of the GHRP-6 on
intact hearts in healthy mice, we decided to evaluate the
effect on a single intraperitoneal dose of GHRP-6 in young
adult rat with intact myocardio on the gene transcription
involved in tissue repair and energetic metabolism. The
study of gene expression, conducted with kinetic analysis
(1-24 hours post inoculation)by means of PCR in cardiac
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YEARBOOK 2013
tissue demonstrated a significant increase in the expression
of its own receptor at 6 hrs in the myocardio. Similarly, a
progressive increase was demonstrated in enzymes related
with cellular metabolism, beta oxidation and mitochondrial
biogenesis.
To study the molecular bases to sustain the effect of the
cardioprotective peptide CIGB 500, the cell line H9C2 was
used in rat’s cardiomyocytos fetal, and protein expression
profiling were identified in four experimental conditions:
1) control; 2) treatment with CIGB 500 (400 μmol, 1 h);
3) treatment with Doxorubicin (1 μmol, 1 h); 4) concomitant
treatment CIGB 500 (400 μmol) + Doxorubicin (1 μmol, 1h).
The study brought about the differential modulation of 111
proteins in the experimental condition #4 by the protective
effect of CIGB 500; which were grouped based on biological
processes significantly represented as: 1) regulation of
the biosynthesis of macromolecules; 2) cascade of signs
intracellular, proteolysis; 3) regulation of protein metabolism;
and 4) transport of proteins. Other eight biological routes
were also identified in the significantly represented 111
modulated proteins different from the control. In 2014, it is
expected to continue with these researches.
Institutions that maintains collaboration:
• National Institute of Endocrinology. Center for the care
of Diabetic Patients.
• National Institute of Angiology and Vascular Surgery.
• Institute of Cardiology and Cardiovascular Surgery.
CIGB-845: Therapeutic combination for
neuroprotection
Dr. C. Diana García del Barco
[email protected]
phone: (53-7) 250 4397
The objective of this project is the research of therapeutic
alternatives facilitating or promoting events of tissue
regeneration in the nervous system after a lesion or wound.
CIGB-845 is a pharmacological combination of a synthetic
peptide and a recombinant protein, which have common
and exclusive neuroprotective effects. It has a wide file
of preclinical trials and safety tests among them, in vivo
experimental studies (experimental autoimmune encephalitis,
axonopathy with 1,2 DAB and global cerebral ischemia).
The in vitro experimental evidences have demonstrated that
this combination exerts protection and recue motoneurons
exposed to pro-apoptotic agents lacking trophic factors.
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YEARBOOK 2013
The prophylactic or therapeutic use of the pharmacological combination
CIGB-845 has induced to significant therapeutic effects compared to
control groups of existing drugs.
The clinical electrophysiological and ultra-structural induction and
characterization in an alternative experimental model of amyotrophic
lateral sclerosis allowed to evaluate CIGB-845 with significant results.
Recently, its potent neuroprotective effects have been demonstrated
in the reduction the volume of cerebral infarction and to rescue brain
cells in an animal model from a global cerebral ischemia. These results
were stably reproducible and are associated with the reduction of clinical
manifestations (Figures 10 and 11). The neuroprotective effect of CIGB845 is similar to the induced effect by hypothermia and superior to the
induced effect by the ischemic pre-conditioning.
Figure 10
volume of the
infarction (mm3)
A
20
15
10
5
×
B
CIGB-428
CIGB-500
CIGB-845
0
vehicle
A) Volume of infarction in animal model of global
cerebral ischemia. Kruskal-Wallis test followed
by Dunn test. The asterisk shows a significant
difference. B) Coronal brain stained with 2,3,5
triphenyltetrazolium chloride(TTC).
False operated
Figure 11
Neuronal density in the cerebral cortex,
caudate putamen nucleus and ventral
nuclei of the thalamus. Representative
images of the caudado-putamen regions:
cerebral cortex and thalamus of the
control groups: false operated ,vehicle
control group and CIGB-845. The black
bar at the right bottom equals to 50 µm.
Cerebral cortex
Vehicle
Caudado-putamen
CIGB-845
Thalamus
Control
Vehicle
CIGB-845
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YEARBOOK 2013
The amount of preclinical tests of CIGB-845 in model
experimental models of multiple sclerosis, lateral
amyotrophic sclerosis and ischemic disease of the nervous
system received the Health annual award in 2011, in the
category of scientific paper.
Figure 12
P8 peptide + IL-15
Interleukin-15
Medium
This project is focused on developing new strategies to
inhibit pro-inflammatory activity of the interleukin-15 (IL-15).
This cytokine is expressed in an uncontrolled form in various
autoimmune and inflammatory diseases such as rheumatoid
arthritis (RA), inflammatory bowel disease, sarcoidosis,
multiple sclerosis and psoriasis. In this context, inhibiting
the signalling induced by IL-15 may be clinically beneficial.
According to this, two strategies have been developed: a
peptide antagonist that binds to the alpha receptor subunit
and a therapeutic vaccine containing IL-15 as antigen. IL15 signals through a trimeric receptor IL-15Rαβϒ and may
be found in a soluble form in biological fluids or in the cell
membrane. Two research projects have been developed for
its inhibition: the development of an antagonist peptide IL15 (project CIGB-55) and the development of a therapeutic
vaccine anti-IL-15 (project CIGB-50).
CIGB-55 is a 10 aa peptide that binds an alpha subunit of the
receptor and inhibits the biological activity in two dependent
cell lines of this cytokine, CTLL-2 and KiT225 (Figure
12). Moreover, this peptide inhibits the secretion of tumor
necrosis factor alpha (TNFα) in synovial cells of patients with
RA (Figure 13) and the induction of inflammatory cytokine
1.0
0,8
0.6
0.4
0.2
Peptide concentrations (µM)
Effect of the antagonist peptide P8 on the proliferation of
KiT225 line induced by IL-15. KiT225 cells were cultured
in presence of 300 pg / ml IL-15 (■), 300 pg / ml IL-15 in
combination with increased concentrations of P8 (♦), medium
(▲). Cell proliferation was assessed by MTT staining..O.D.
optical density.
Figure 13
180
160
140
TNFα (pg/mL)
O.D. 576 nm
Dr. Alicia Santos
[email protected]
Phone: (53-7) 250 4486
120
100
80
60
αTNF secretion is inhibited by the IL-15 antagonist peptide.
Synovial fluid cells were incubated with peptide or peptide
plus IL-15 in 24-well plates and cultured for 24 h. αTNF levels
were measured by specific ELISA.
40
20
Cells
Cells + peptide
Cells + peptide + IL-15
26
YEARBOOK 2013
IL-6 and IL-8 produced by the mechanism of inverse signal
through IL-15 of membrane. Currently, the effectiveness
of CIGB-55 is being evaluated in animal models, and it's
being working in modifying the peptide and developing new
formulations to improve its stability. This project has a patent
granted in the USA, The European Union, Australia, Canada,
Russia, Mexico and South Africa.
Another strategy consists in the active immunization with
IL-15 structurally modified in a mutein of IL-15 not active,
using aluminium hydroxide as adjuvant. The experimental
results of the immunization in non human primates, showed
the induction of high titers of neutralizing antibodies against
a human IL-15 not modified (Figure 14). An anti-cytokine
therapeutic vaccine may be advantageous with respect to
the use of monoclonal antibodies against IL-15 , because it
involves less frequent injections per patient (2-4 per year),
no response against antibodies that it is one of the causes of
loosing response to the treatment; also its low cost.
O.D. 576 nm
Figure 14
2.5
Pre-immune serum
2.0
Serum from immunized
monkeys L-15
1.5
Medium
1.0
IL-15
0.5
0
1
100
10 000
1 000 000
Dilutions of serum
Effect of serum from immunized animals on the proliferation of CTLL-2 line,
induced by IL-15. CTLL-2 were cultured in presence of 300 pg / ml IL-15
(■), 300 pg / ml IL-15 plus serial dilutions of serum (♦), medium (▲). Cell
proliferation was assessed by MTT staining.O.D. optical density.
CIGB-50 is in its developing stage of scale-up of the protein
production. The preclinical studies are in course. This project
has a patent granted in The USA, the European Union and
China.
At the same time, basic studies are conducted to characterize
the pattern of pro-inflammatory cytokines and to introduce
IL-15 and its receptor in synovial fluid of patients with RA.
27
YEARBOOK 2013
Identification of HIV inhibition target
cells of VIH-1
Dr. Celia Fernández
[email protected]
phone: (537) 250 4396
Immunological reconstitution of the immune system is
essential in the development of therapeutic strategies for
effective clinical management of patients with different
conditions.
The immunological reconstitution of the immune system
is essential in the development of therapeutic strategies
for the effective clinical control of patients with different
diseases. Dialyzable leukocyte extract (DLE) is a material
of low molecular weight coming from the rupture of human
peripheral leukocyte, consisting of a large number of
molecular entities like the transfer factor (TF) which is
the entity responsible for transferring specific antigen
delayed hypersensitivity. DLE is used in the treatment of
viral, bacterial, parasitic and fungal infections, as well as
immunodeficiency, neoplasia, allergies and autoimmune
disorders. In recent years, the use of DLE reported in
individuals infected with human immunodeficiency virus
VIH-1. In a clinical trial in Cuba in which DEL was used,
a significant delay was shown in the onset of symptoms
of the disease in asymptomatic seropositive patients with
VIH-1 comparing not treated patients.
The most current effective therapy known as the highly active
antiretroviral therapy (HAART), provokes a suppression in
viral levels in blood and improves the function of the immune
system in most of the patients. The normalization of the
immune system and the total elimination of VIH-1 have been
not proven yet, though.
HAART is also involved in potential toxicity as well as the
emergence of viral strains resistant to treatment. Another
difficulty of the therapy is related to its high cost and the
lack of resources in many countries with the highest rates
of infection. Disadvantages associated with HAART have
motivated the research of other therapeutic agents for the
treatment of VIH-1. Recent researches explore the pathways
to inhibit the replication of VIH-1 using cellular factors as
target, which depend on the virus replication, including
receptors and co-receptor molecules of VIH, transcriptional
and assembly factors, among others. The identification of
cellular factors inhibiting the viral replication cycle is one of the
most important goals in the researches related to anti-VIH-1.
It is stated that a therapeutic strategy that makes good use it
would have advantages or take multiple benefits specially, the
possibility to reduce the appearance of viral resistance.
This project aims to elucidate the effect of DLE on virus
replication of VIH-1 and the host mechanisms influencing in
its immunopathogenic process. By using an in vitro system
for studying the MT4 human cell line, it was possible to
demonstrate a dose-dependent inhibitory effect on HIV-1
replication in cells treated with DLE before and after the viral
challenge for 24 hours or more (Figure 15).
Figure 15
B
100
Percentage of inhibition (p24)
Percentage of inhibition (p24)
A
80
60
40
20
0
0.15
0.3
0.6
1.25
2.5
100
80
60
40
20
0
0.15
0.3
DLE (U/mL)
0.6
1.25
2.5
DLE (U/mL)
Dializable leukocyte extract (DLE) effect on HIV-1 replication in vitro. RT: reverse transcription.
28
YEARBOOK 2013
Moreover, the in vitro DLE reduces the production of TNFα
induced by LPS and the activity of the transcriptional factors
NFκB and Sp1, necessary for the expression of the viral
genome (Figure 16).
Figure 16
Controls 0h
1
2 3 4 -
3h
3d
7d
- 1.2 2.5 - 1.2 2.5 - 1.2 2.5
Time
DLE
Protein DNA complex
Free oligonucleotide
Inhibitory effect of DLE on the
binding activity of transcriptional
factor NFkB.
Researching endogenous factors involved in the
immunopathogenic process not known so far, the project
has developed a comparative proteomic study and identified
an important number of proteins. The project is validating the
function of proposed proteins as therapeutic targets on the
HIV infection, also designing and evaluating new molecules
that act or imitate the effect of the proteins to achieve viral
inhibition. The RNA technology of interference is used to
study the protein function in the infection process with HIV-1.
The project has designed a group of molecule candidates for
a vaccination against HIV-1; and one of them with a potent
antiviral activity.
Department of Diagnostics and
Genomics
Head f the department
Dr. Lidia Inés Novoa
[email protected]
phone: (53-7) 208 4035 (Center of Biological Investigations)
The Department of Diagnostics and Genomics is focused on
pharmacogenomic studies by means of the ethnic mixture
and the organization of the National Health System in Cuba,
29
YEARBOOK 2013
along with the potential of new developing products in clinical trial phases
in the projects against cancer, autoimmune, chronic and infectious
diseases. The CIGB counts with a technological platform of microarrays
from the Chinese company Capital BIO (Figure 17). This technological
platform is certified by ISO 9001, ISO 13485, the FDA (Food and Drugs
Administration), the European Community, and the regulatory authorities
of China.
Figure 17
RT-Cycler
SmartArrayer 48
Lux Scan 10K-A
Nano drop
Slidewasher
Extractor 36
Biomixer II
Complete platform of double fluorescence microarray.
Farmacogenomics
MSc. Daniel Palenzuela
[email protected]
Phone: (53-7) 208 7421 (Center of Biological Investigations)
The group is in charge of pharmacogenomic study in projects of generating
new biopharmaceuticals products or vaccines and clinical trials of those
products against cancer, autoimmune and infectious diseases:
Objetives in course
• Study of the mechanism of action of Heberprot-P® and sampling of
Phase IV clinical trial.
• Identification of new thrapeutical targets in the treatment of the
diabetic foot ulcer.
30
YEARBOOK 2013
• Study of markers of response to HeberFeron®
• Study of mechanism of action and markers of response
of CIGB-300 in different pathologies
Identification of new biomarkers and therapeutic
targets in the UPD
The database analysis of 49 gene expression, determined
by qPCR, was complimented by a methodology that allows
its analysis to a level of each biological replication. This
procedure permits the application of methods multi-varied
of analysis as: two-dimensional groupings (Figure 18)
to visualize the global behavior of studied samples and
gene profiling, analysis of the main components for gene
determination and relevant biological events also the use
of other confirmatory methods on differential expression of
these genes that finished with the evaluation of its predictive
ability as a preliminary stage for its posterior validation. As
a result, a group of some genes were identified that can be
predictors of response to the treatment with Heberprot-P®
distinguishing ischemic and neuropathic cases or both as
a whole.
Figure 18
-0.017030612
0.9122645
0.9403377
0.4404062
0.05952531
IsqResp T0 38
IsqResp T0 39
IsqResp T0 20
IsqResp T0 47
IsqResp T0 69
IsqResp T1 47
IsqResp T1 39
IsqResp T1 38
IsqResp T1 20
IsqResp T1 69
NeuResp T0 5
NeuResp T0 12
NeuResp T0 13
NeuResp T1 11
NeuResp T1 12
NeuResp T0 11
NeuResp T1 5
NeuResp T1 13
NeuResp T0 14
NeuResp T1 14
IsqNoResp T0 1
IsqNoResp T1 98
IsqNoResp T0 61
IsqNoResp T0 98
IsqNoResp T0 95
IsqNoResp T1 1
IsqNoResp T1 61
IsqNoResp T1 95
IsqNoResp T0 133
IsqNoResp T1 133
IsqNoResp T0 10
NeuNoResp T1 10
NeuNoResp T0 16
NeuNoResp T1 16
NeuNoResp T0 88
NeuNoResp T1 88
NeuNoResp T0 20
NeuNoResp T0 27
NeuNoResp T1 27
NeuNoResp T1 72
0
Two-dimensional groupings to visualize the global behavior of studied samples and gene profiling.
31
YEARBOOK 2013
Effect of the epidermal growth factor in tumor cell
lines A431 in mouse: in vivo analysis inhibiting of
the tumor growth and gene expression
The results evidenced safety using EGF as active ingredient
of formulations in clinical trials to nanomolar concentrations
which prolongs life of the xenotrasplanted mice models with
the tumor cell A431 inhibiting tumor proliferation. Findings
in the differential expression to those tumors, after the
treatment with EGF and compared to placebo reinforced
those phenotype observations; and the most important
genes, that were differential expressed, were TP53, CDK4
and CASP9 (Figure 19); favoring inhibition of the cell cycle
and the apoptosis.
In the in vitro study of the EGF’s effect on cell growth, using
nanomolar concentrations, a statistically significant reduction
Figure 19
A 300000
Cells/mL
165 nM
33 nM
2.2 nM
250000
200000
150000
100000
50000
0
EFG
Placebo
EFG
Placebo
EGF
Placebo
Not treated cells
Treatments
B
placebo
C
treated
EGF
EGFR
350
ERBB2
Tumor volume (mm3)
300
PI3K
250
RAFA1
AKT1
MYC
200
CASP9*
150
CDKN1A/P21
CDK4*
P53*
100
INHIBITION OF
50
CASP1
0
1
2
3
Days
4
5
6
7
ACTIVATION
THE CELL-DIVISION
CYCLE
OF APOPTOSIS
Analysis of nanomolar concentrations of EGF effect on cell proliferation A431. A) Effect of different concentrations of the human
recombinant epidermal growth factor (hrEGF) in A431 cell proliferation in culture, compared to non-treated controls. B) Total volume
of A431 tumors implanted in naked NIH mice treated either with hrEGF or placebo. C) Diagram of some of the signaling cascades
being activated in the A431 cells which were implanted in naked mice and treated with nanomolar hrEGF concentrations.
32
YEARBOOK 2013
Tabla 1
Human cellular lines Cancer
Origen
Levels of
rhEGF
K562
Erythroleukaemic cell line Chronic Myelocytic
Leukemia
Ccl-2433
(lymphoblastic)
0.6 %
Mcf-7
Breast cancer cell line. Mammary gland
adenocarcinoma.
Htb-22 (epithelial)
1.2 %
U1906
Small cells of lung cancer (SCLC).
Epithelial
6.5 %
Ls-174t
Human colon adenocarcinoma
Cl-188 (epithelial)
54.2 %
A431
Human epidermoid carcinoma cell line. Skin
Squamous Cell Carcinoma
Crl-1555
(epithelial)
99.3 %
Nci-h125
Non-small cell lung cancer (NSCLC)
Epithelial
99.1 %
Effect of the recombinant human epidermal growth factor (rhEGF) in tumor cell lines with different cell lines used in
the in vitro study of gene expression with rhEGF.
of cells was achieved with high and medium levels of EGF
receptors (H125, LS-174 and A431) (Table 1).The analysis
of the results evidenced that these genes influenced
predominantly in the process of the cellular cycle inhibiting the
cell multiplication through a cycle arrest in the phases G1-S.
Department of Physical-Chemistry
Dr. Luis Javier González
[email protected]
Phone: (537) 271 6022, ext. 1122
The identification of molecules functionally involved in
specific diseases and resistance to drugs, with application in
bioinformatics and proteomics, protein engineering and protein
modelling for the development of new drugs, are the issues
of the Department of Physical-Chemistry. The department
has experience in purification of proteins, characterization
of glycosylated proteins, the synthesis of peptides and
oligonucleotides, gas chromatography, study of the interaction
protein-protein, ultrastructural analysis electronic microscopy,
mass spectrometry and amino acid analysis and imaging
analysis obtained through nuclear magnetic resonance.
Projects in course
• Proteomics.
• Bioinformatics.
• Screening virtual.
• Chemical modifications of proteins.
• Development of modified peptides (APL) with applications
•
•
in the treatment of artritis rheumatoid arthritis and other
autoimmune diseases.
Development of molecules with antiviral activity against
dengue virus.
Design of antiviral molecules against VIH-1.
Development of methodologies for the
study of proteomes
Dr. Vladimir Besada
[email protected]
Phone: (537) 250 4148
The development of methodologies for the study of proteomes
aims to develop the most complete characterization of
proteomes. In preceding years, we have been working on
some methods of selective isolation of peptides allowing
the simplification of complex mixtures to make quantitative
proteomes without using two-dimensional electrophoresis,
and with the isotopic labeling allowing the quantification
of protein differential expression. The proposed method is
based on the three known methodologies with the acronyms
SCAPE (Selective Capture of Peptides). This method allows
the isolation of peptides with different charges: neutro, with
one or two, described as RH0, RH1 and RH2, respectively.
The recovery of the selectively isolated peptides is over 70 %
in all cases; the selectivity is greater than 95 %, and a wide
33
YEARBOOK 2013
coverage can be achieved (greater than 95 %) of the known
proteomes. The fractioning, achieved by ion exchange by the
use of certain reactive blockers of the amino groups allows the
identification of a greater number of proteins. We demonstrated
that with only one methodology is possible to isolate the three
groups of peptides in concern An alternative method is the
so-called DF-PAGE, based on the electrophoretic fractioning
at protein level in the first dimension, later on at peptides level
as second dimension. There are other strategies of work like
the fractioning by reverse phase to acid and basic pH, and the
use of peptide libraries to equalize complex protein mixtures.
These methods allow the relative quantification of proteins
from two to four conditions simultaneously
These methods have been used in several projects of the
direction of biomedical researches with higher results than
those obtained by the two-dimensional electrophoresis for
instance: the elucidation of mechanisms of action of some
pro-apoptotic and antitumor molecules, the study of arthritis
rheumatoid in biomodels , and the protein identification in
complex mixture of proteins. We began to validate targets
from the results of previous projects that could conduct
experiments of proteomics hypothetically directed by means
of Single Reaction Monitoring.
Additionally, quality control of recombinant proteins and
synthetic peptides produced at the CIGB and other Cuban
scientific institutions are achieved through techniques of
mass spectrometry, protein electrophoresis, liquid and gas
chromatography, and amino acid analyses; complying with
Good Laboratory Practices. These techniques could also
conduct stability tests and the most complete characterization
of some products.
In this department, a project of autoimmunity has been achieved
the design of altered peptide ligand (APL), that modulates the
response of circulating autoreactive clones in the periphery
which have potentialities for the treatment of autoimmune
diseases like RA. The most advanced candidate peptide other
APLs have patents; and in 2013, a phase I clinical trial of one of
these peptides was approved to begin in 2014.
Autoimmunity
Dr. María del Carmen Domínguez
[email protected]
Phone: (53-7) 250 4387
The project of Autoimmunity is focused on the study and
research of treatments of autoimmune diseases, specifically
of RA. This is a systemic autoimmune disease, whose primary
clinical manifestation is the inflammation of the peripheral joints.
This disease affects 1 % of the population worldwide, with a
woman/man rate of 3:1. In the pathogenesis of the RA, several
cell populations are involved that enhance and perpetuate the
disease like: T and B lymphocytes, microphages, neutrophils
and its products. The social and economic impact of the RA
is substantial, not only for countries but also patients: the
medical and surgical treatment, disability induced in workingage people cause costs of millions of dollars.
Traditionally, RA has been treated with drugs that tend to
improve symptoms or the use of powerful nonspecific
immunosuppressive not constituting an efficient therapy.
However, the advances in the knowledge of pathophysiology
of this disease and biotechnology have led to the emergency
of a new era in the treatment of the RA, known as biological
therapy, in which it has attempted to block several key
elements in the pathogenesis of the disease. Among those
therapies, there are different anti-TNFα drugs approved
by the Food and Drugs Administration (FDA) that currently
constitute the most successful alternatives for the treatment
of RA. Although the good results after the use of the
products, about 50 % of patients do not get better with this
type of therapy because those drugs can cause suppression
of the effector mechanisms of the immune response at a
systemic level; a lot of patients present a high susceptibility
to infections, other autoimmune diseases and neoplasia;
therefore it is not an effective therapy for the disease.
The development of therapeutical alternatives that can get
rid of specific pathogenic cells without causing generalized
immunosuppression is a challenge today. In recent years,
this has brought about for some researches to work with
the use of antigen-specific strategies for the treatment
of autoimmune diseases, trying to regulate the immune
response, rather than suppress it. Peptides derived from
auto antigens or altered peptide ligand (APL,) have been
used administrated in conditions that allow the induction of
regulatory mechanisms mediate the induction of peripheral
tolerance (anergy, apoptosis and induction of T cells).
This research project is precisely focused on the development
of specific- antigen strategies for the treatment of RA aiming
at the obtainment of effective drugs for the treatment of this
disease.
The strategy developed is based on the induction of
peripheral tolerance, through the use of a modified APLtype peptide, and derived from autoantigens involved in the
pathogenesis of the RA. The autoantigens selected is the
protein from heat stress of 60 kDa (HSP60).
34
YEARBOOK 2013
In general, the HSP60 have immunoregulatory properties that
help the maintenance of the tolerance; that´s why are strategic
targets in the development of therapies for the treatment of
inflammatory disorders like RA.
From the sequence of the human HSP60, a new peptide
fragment was identified as a possible epitope of T cells
through bioinformatic tools. This epitope was the bases for
the design of an APL- type peptide.
The APL are immunogenic peptides ,analogue, with one or
several substitutions at essential position contact with the
TcR or with the major histocompatibility complex (MHC),
which interfere or modify the cascade of required events
to complete activation of T cells (Bielekova B, Martin R. J
Mol Med. 2001;79:552-65). The possibility to manipulate
experimentally las intrinsic properties of known peptide
ligands peptidases allow to alter appropriately nature, course
and strength of the cellular immune response.
The APL - type peptide designed by our group through
bioinformatic tools was modified in a aminoacid residue in
relation to the original epitope of the HSP60 (E18-3), to increase
affinity and the probability to be present by the human leukocyte
antigen (HLA) molecules class II that is more frequently
expressed in patients with RA. This peptide is called CIGB-814.
The CIGB-814 peptide exerts a potent therapeutic effect
in two animal models for the RA: arthritis induced by an
adjuvant (AA adjuvant arthritis) (Figure 20) and arthritis
induced by collagen. Moreover, it also induces regulatory
cells with a phenotype CD4+CD25highFoxP3+ in ex vivo
trials with mononuclear cells isolated from peripheral blood
and synovial liquid of patients with RA (Figure 21).
Figure 20
Clinical signs AA
Group 1
18
Group 2
16
Group 3
14
Group 4
Group 5
12
10
8
6
4
2
0
10
20
30
40
50
Posterior days of the AA induction
Evaluation of clinical signs in rats with arthritis induced by adjuvant and treated with peptides: original (E18-3) and CIGB-814
(APL1). On the tenth day of the disease induction, the rats were randomly divided in four treatment groups : group 1: rats
inoculated with the E18-3 peptide by intradermal route; group 2: rats inoculated with APL1 by intradermal route; group 3: rats
inoculated with APL1 by subcutaneous group; group 4: group placebo; group 5:healthy rats. The different values of the clinical
signs corresponding to each day represent the average per group plus the standard deviation. The graph represents the results of
one of the three experiments. The statistical analysis between groups was performed with the parametrical testANOVA and means
were compared using a posteriori Tukey test. Different letters mean significant statistical differences at p < 0.0001.
35
YEARBOOK 2013
% of Tcells CD4+CD25highFoxp3+
Figura 21
6
a
4
a,b
b
2
0
APL1
C-
E18-3
Treatments
Analysis of the induction in rheumatoid arthritis patients' PBMCs cells in culture of the Treg CD4+CD25highFoxp3+ phenotype,
after its incubation with the altered peptide ligand 1 (APL1). Results are compared to those of cultures incubated with the native
peptide or without peptide stimulus. PBMCs were stimulated for five days with 40 mg/mL of the CIGB-814 peptide (APL-1) or the
native peptide (E18-3). Baseline stimulation levels were set from PBMCs without peptide stimulation for each patient (C-). Groups
were compared by a parametric ANOVA test and means by a posteriori Tukey's test. Results are presented as means plus/less
standard deviation. Different letters stand for highly statistically significant differences (p < 0.0001).
Development of molecules with antiviral
activity against dengue virus
BSc. Glay Chinea
[email protected]
Phone: (537)364
Dengue virus (DENV) is a flavivirus transmitted by mosquitoes
which constitute a human disease, with an important global
impact in public health and economy. The infections due to this
virus cause a wide spectrum of clinical manifestations, some
asymptomatic and benign like dengue fever, characterized
for an undifferentiated fever, and other severe as the dengue
hemorrhagic fever (DHF) and the potentially lethal dengue
shock syndrome (DSS). During the last six decades, el DV
has gradually increased its geographic distribution and the
number of infected patients. Nowadays, it is pandemic in
tropical areas: from 50 to 100 millions annual infections are
estimated to occur and 2500 millions of people live at risk of
being infected. The average annual of cases of FHD reported
to the World Health Organization is up to 500 000 per year;
among them, more than 20 000 correspond to fatal cases.
Nevertheless, the real load of the disease is considered to be
still several times higher. Despite of this, vaccines or specific
antiviral treatments are not currently commercialized.
The project of developing antiviral agents against DV, at the
CIGB, is in charge of the study of different stages of virus
replication, attractive for the identification of new principals
for the inhibition of the disease. A key strategy of the project
is focused on the inhibition of virus entry into target cells; a
process involved in a series of early events of infection, like
adherence of the virus to the host cells, internalization or
endocytosis to cells and the fusion of membranes in early
endosomes. Another attractive target is processing the
viral polyprotein by the NS3 protease, through the design
36
YEARBOOK 2013
of molecules that interfere in the formation of the complex
process of maximum proteolytic activity NS3/NS2B.
Currently, some available molecules of protein nature (also
peptidic and drug-type) show antiviral activity in vitro and
in vivo, in the model of induced encephalitis, by intracranial
inoculation of DENV in mice. These molecules bring about
new principles of inhibition, protected by four patents
(granted and requested or applied).
Identification of putative endocytic receptor LRP1
as dengue virus: a new strategy for the development
of the antiviral therapies
The DV penetrates in the mammalian cells by receptormediated endocytosis. Other research groups have shown
that several molecules of the cell surfaces bind the virus,
such as: GAGs, DC-SIGN, HSP 70, HSP90, laminin
receptor, and macrophage receptor. Receptors also bind FC
gamma DENV as immunocomplexes. It has previously been
proposed that these molecules are virus receptors; however,
none of them behaves like endocytic receptor of the DENV
and their roles seem to be more related to initial events of
adhesion prior internalization of the virus.
Concerning this, we have isolated and identified the LRP1
receptor as endocytic putative receptor of the DENV. Our
studies are focused on the characterization of the LRP1
binding to the virus and the envelope protein (protein E),
determining the function of LRP1 of the virus and protein E
internment like in in vitro and in vivo assays inhibiting the
infection of LRP1 preparations and their ligands.
A patent was granted concerning the use of the LRP1 as a
potential target for the development of antiviral agents. The
target’s blocking would inhibit the virus’ entry which leads to
a productive viral infection, regardless the occurrence of the
virus’ binding to other adhesion receptors.
Recombinant fragments of LRP1 have also been expressed
in order to establish binding assays to identify drugtype molecules by in vitro screening compound libraries.
Moreover, in silico screening of virtual libraries of chemical
compounds have been studied and a number of them for in
vitro screening of potential inhibitors of virus binding to the
ligand binding domains of LRP1 have been selected.
Design of inhibitory molecules penetrating the
virus to the cells
Based on the domain III structure, a number of constricted
peptides were first designed to imitate a topographic epitope;
and those synthetic peptides show an antiviral activity
against DV. One of them inhibits the in vitro viral infections
DV1-4 with IC50 from 15-50 μM and a selectivity rate of
20-67. This peptide shows a significant protection in an
encephalitis mice model caused by intracranial inoculation.
Moreover, it was demonstrated that the peptide binds the
LRP1 receptor fragments using the Plasmon resonance
methodology (BIACORE). The identity of the peptide and its
variants constitute an intelectual property of the CIGB whose
patent has been granted in some countries Currently, we are
working in the obtainment of more potent variants with IC50
values at the nanomolar level.
On the other hand, starting from the in silico results of our
in silico screening studies in virtual libraries of chemical
compounds against LRP1 receptor, a group of potential
binding compounds were obtained which were tested
in relation to the binding of the receptor fragments by
BIACORE. Some compunds have been identified and are
the object of characterization in biological trials and the
design of modifications for the optimization of properties.
Protein E as target for the development of antiviral
agents
According to the functional importance of the E protein,
it was also selected as a target for the development of
ligands able of blocking the sites involved in protein-protein
interactions, protein-membrane interactions and interfering
in conformational changes of the protein. A fragment of
recombinant single chain antibody is an antiviral agent
that is developing with neutralizing capacity against the
four serotypes of the virus. Drug-like molecules have also
been identified against this target, using virtual screening
methods.
This group has developed a library of Alanine mutants of the
exposed residues of domain III in order to identify regions of
interaction of the envelope protein (domain III in particular)
monoclonal antibodies, sera from patients and cell receptors.
This tool has allowed to identify immunodominant epitopes
of the human humoral response and essential residues
in the interaction with receptor proteins. In this project, a
combination of methodological tools have been used like:
the computational design of molecules based on structure
and virtual screening, the selection of ligands against
target proteins using combinatory methods(combinatory
libraries of phages) methodologies of proteomics, methods
of analyses of biomolecule interactions, among others. In
addition, in vitro and in vivo trials of the antiviral activity are
37
YEARBOOK 2013
also implemented for the evaluation of candidate molecules
and the validation of the putative viral receptor.
The crystal structure of the 4G2 monoclonal antibody was
solved. Modelling studies on the recognition region led to the
understanding of the structural determinants of the broad crossreactivity of antibody against flavivirus, and their mechanism
of neutralization and generation of escape mutants.
Survey of chemical compounds in virtual libraries
for the development of new drugs
MSc. Osmany Guirola
[email protected]
Phone: (53-7) 250 4150
The use of different computational procedures for drug
design has emerged as a need to accelerate and streamline
the development of new drugs. The success of this approach
will depend on the close link between biology groups who
know the target deeply, and the function upon which is to
influence, the group of bioinformatics associated to the
rational design, and the group of chemical synthesis
The survey in virtual library is a computational procedure
evaluating the possibility of specific interaction of a
relatively small organic molecule with the desired protein
site. Possibilities of interaction for all compounds listed in
databases of chemical compounds are compared, which
can generally be integrated by several million compounds
An alternative, or rather a possible combination, are the de
novo design methods that are not based on a database of
known chemical compounds, but try to explore the extremely
broad chemical space (which is considered 1060 to 10100
compounds) that would constitute a drug. Some studies
have been conducting to obtain antivirals against Dengue
virus. There are two strategies: one inhibiting the virus fusion
with the cell membrane and the other blocking the interaction
between the virus and its cell receptor. This receptor was
identified by studies of proteomics as the involved in the
internalization of such virus to the cell.
One of the main causes of failure of the development of drugs
is not take into account the ADME/TOX properties during
the initial stage of development. As part of the strategies to
develop the techniques of this survey, some methods have
been developed using SVM to predict ADME/TOX properties
of the possible antiviral candidates. With the amount of
predictors of ADME/TOX properties developed, greater
probabilities of success of the future drug may be guarantied.
Chemical Synthesis
Leader of the Laboratory
MSc. Osvaldo Reyes
[email protected]
Phone: (537) 250 4537
The work of the group Chemical Synthesis is focused on two
fundamental lines: synthesis and purification of oligonucleotides
and modified oligonucleotides, and the design and obtainment
of synthetic peptides for different applications. We are also in
charge of the design and obtainment of combinatory libraries
of peptides to find peptides with therapeutical properties and
the study of the metabolic stability of some of those molecules
in biological fluids as basis for the selection and optimization
of therapeutic candidates of peptide nature.
The group of oligonucleotides synthesis, the only one of
its kind in Cuba, satisfy the demands of these molecules
at the CIGB and other scientific centers of the country. It
also serves as a national command post for contingencies
that require medical services. So far, more than 60 000
oligonucleotides have been synthesized increasing every
year the total amount of synthesized molecules.
According to the synthesized peptides, more than 2000
molecules have recently been synthesized. The group’s
current capacity includes the synthesis of few milligrams
and of even tenths of grams of any aminoacidic sequence.
Various chemical modifications are also achieved to peptides,
among them: the binding cycling by disulfide and lactam
bridges, modifications to the amino groups, conjugation to
other macromolecules, polymerization by disulfide bonds
and the obtainment of complex structures such as multiantigenic peptides (MAP).
The development of possible therapeutic candidates based
on synthetic peptides, as well as the study and process
design for the production scale up of these molecules are
also part of the objectives of this group.
Characterization of N- and O-glycans bound to
glycoproteins
[email protected]
phone: (53-7) 250 4366
Glycobiology is a field of biology in charge of the study of
carbohydrates and its influence in cell functions with relevance
to biochemistry, cellular and molecular biology, histology and
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YEARBOOK 2013
immunology studies among others. Carbohydrates, which
may be present as an extraordinary structural diversity and
during decades they served like energy sources (glucose and
glycogen) and as structural components; but now it is known
that its structures contain information that modulates directly
many cellular processes. They can be used like molecules of
direct information, transmitting messages between the cells
or inside them. The exchange of carbohydrates molecules
can affect the structural transformations or properties of
cellular union influencing in the functioning of the immune
system, the operative capacity of several infectious agents
and cancer progression (Gabius, 1997).
According to cancer, the selectins provide the best example
of the interaction lectin-carbohydrates in the biological
recognition. Selectins also have biological properties of
disseminating carcinogenic cells from an initial tumor to
other places of the organism and that the blockade of union
places may help prevent metastasis forming. Additionally,
galectins (soluble lectin with affinity for galactosides) can
act as modulators of cells - substrate interactions essential
for the normal differentiation and growth of all multicellular
organisms. They are capable of inducing cell proliferation,
cell death or apoptosis and has been involved in the organs
morphogenesis, metastasis of tumor cells, leukocyte
trafficking, immune response and inflammation, as well as
the extracellular matrix recognition (Sharon y Lis, 2004).
These findings in Glycobiology have opened a way of
investigation in the research of glycomarkers which
allows to diagnose and evaluate the patients’ progression
suffering from carcinogenic diseases as cancer of prostate
that occupies the second most frequent type of malignant
neoplasia in men. More than 650 000 new cases are
diagnosed every year in Cuba in 2012. Prostate Cancer
provokes 2703 deaths (MINSAP. data, 2012).
The prostate-specific antigen measurement is considered to
be one of the best biochemical markers currently available in
the field of oncology for prostate cancer (CaP) and permits
the early detection of the pathology and the patients’ followup (Stamey et al. 1987).
High PSA levels in sera, mostly produced as a consequence
of a rupture of the prostate membrane by cancer cells, are
indicatives of the CaP presence (Brawer, 1999; Laguna and
Alivizatos, 2000; Milford Ward et al., 2001). However, the level
of this protein allows to distinguish cellular changes caused by
cáncer or by benign changes in the prostate (BPH). As a result,
the PSA tests have a high rate of false positives, that could
mean to repeat biopsies and unnecessary surgeries (Laguna
and Alivizatos, 2000; Stamey et al., 2002; Bonn, 2002).
Due to a limit specificity of the routine PSA, it has been
necessary to develop additional procedures, more
informative, in CaP cases using also blood as a starting
point for the analysis.
In the last years, researches have been focused on the use
of glycoproteomic techniques to define biomarkers of CaP.
As a result of these studies, 14 proteins were identified as
potential serological biomarkers to detect CaP with different
specificity and variable selectivity, in which 11 proteins
presented potential N-glycosylation and O-glycosylation
sites (Adam et al., 2001; Banez et al., 2005; Pennington et
al., 2005; Ornstein et al., 2006).
Most of the studies have been conducted from serum where
there are a considerable number of glycoproteins and
require analytical methodologies of high sensitivity allowing
to establish differences between glycosylation patterns in
healthy and sick individuals.
In 2013, using the combination of analytical methodologies
that include the isolation of N-glycans marked with
2-aminobenzamina
in
Amide-80
Normal
Phase,
exoglycosidases digestion, monosaccharide analysis
and mass spectrometry, it was achieved changes in the
N-glycosylation patterns of 44 patients with prostate cancer
included in a phase II clinical trial with a therapeutic vaccine
candidate. The results showed a decrease of glycosidic
structures related to the progression of this disease.
Laboratory of Electronic Microscopy
Dr. Viviana Falcón
[email protected]
Phone: (537) 271 6022, ext. 1127
Transmission electron microscopy is an indispensable
analytical tool in the detection of viral particles or other
condition by in vitro and in vivo, both preclinical and clinical
trials. The nanoparticles obtained through recombinant
DNA technology, have been used extensively to the
development of vaccine candidates against viral diseases.
These nanoparticles mimicking supramolecular structures of
viruses and induce an antiviral immune response, based on
neutralizing antibodies and protective cellular response. The
paradigm of these recombinant nanoparticles is hepatitis B
vaccine whose use has been widespread in humans. The
HBsAg is assembled into a nano proteolipoparticule, which
generates a protective immune response of long duration.
The list of nanoparticles developed by the CIGB, currently in
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YEARBOOK 2013
preclinical and clinical studies, also includes other vaccine candidates for
hepatitis C, dengue, hepatitis A and RHD (Figure 22). The nanoparticles
can be formed by only proteins or protein-lipid complexes and proteinDNA. The expression systems used are the bacterium Escherichia coli,
yeast Pichia pastoris and leaves of plants of snuff. Most viral capsids
or nanoparticles are mimicking the lipoprotein envelope of enveloped
viruses. Using this valuable tool, the quality control of various production
processes can be achieved.
This technique is also used in the detection of viral diseases in different
industries like: agricultural, fisheries and veterinary.
Figure 22
Subviral Nanoparticles obtained by the recombinant DNA technology.
A) Presence of the HBsAg nanoparticles is detected of 20 to 30 nm in
diameter. B) Presence of HBsAg nanoparticles is detected of 20 to 30 nm.
C) Presence of HBsAg + HBcAg nanoparticles is detected of 20 to 30 nm
in diameter. D) Presence of 50 nm HCAgc nanoparticles is detected. E)
Presence of the nanoparticles VP60 from 35 to 40nm calicivirus is detected.
F) Presence of nanoparticles of the DIIIC VHA-ADN con 27nm de diameter
is detected. G) Presence of nanoparticles of the de 50 nm del dengue-2/
AND is detected. H) Presence of nanoparticles of 50 nm of capsids- DomIII
dengue/DNA is detected. Bar = 100 nm
Group of Imaging
Dr. Carlos Cabal Mirabal
[email protected]
phone: (537) 250 4620
The group of Imaging deals with the implementation of different imaging
methods for molecular, preclinical, clinical and pharmaco-surveillance
studies. In particular, it develops procedures of magnetic resonance
imaging (MRI) to quantitative evaluation of pharmacokinetics and
biodistribution; also the effectiveness of medications and new formulations
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YEARBOOK 2013
using classical imaging of magnetic resonance imaging
(MRI) weighed up in density T1, T2 as well as imaging
weighed up of diffusion, perfusion and in vivo spectroscopy
of MRI.
This group has obtained relevant results as follows:
• Development and validation of a method for determining
the pharmacokinetics and in vivo biodistribution in
biomolecules marked with magnetic particles from
magnetic resonance imaging (MRI) in animal models.
It deals with a new method of quantitative determination
of the pharmacokinetics and in vivo biodistribution with
magnetic resonance imaging (MRI) of biomolecules
marked with magnetic particles. The method is valid for
magnetic nanoparticles functionalized with peptides,
proteins, monoclonal antibodies or other biomolecules
determining 1) the concentrations for compartments, 2)
the time of residence, and 3) the speeds of absorption
and excretion. The method allows the animal model’s
follow up without sacrificing but comparing its temporary
evolution, which makes the studies more rapid and
economic. It has been presented in events and published
in magazines of high visibility.
• Establishment and validation of new biomarkers and
protocols of magnetic resonance imaging to evaluate
solid tumor responses under therapeutic action of
medications. It was established a protocol for the in
vivo, quantitative and evolutionary study of solid tumors
from magnetic resonance imaging in animal models and
human beings that allow to evaluate the evolution of
tumor volumes, morphology changes, different internal
textures of the tumor micro - regions, changes in cell
density and metabolic activity under the medication
action. In particular, 35 pediatric patients were studied
during 3 years using the treatment with Nimotuzumab
and with the tratamiento suspended. In addition the
parameters and magnitudes in mice were also studied.
It has been presented in events and published in
magazines of high visibility.
allows monitoring the interaction between two molecules
in real time. Some of the potential applications include
measurement of affinity interaction and binding kinetics,
the determination of analytes and epitope mapping. The
principle of detection based on SPR is an optical technique
measuring the variations in refraction index at the sensor’s
surface, caused by the differences in analyte concentrations.
Hence, the technique does not require molecule marking to
be analyzed and the response is essentially independent of
the bio-molecule’s nature.
This technology can be used in the study of protein
interactions, nucleic acid, lipid micelles and even particles
like viruses and whole cells. Application areas go from basic
science in biochemistry and molecular biology, to the design
of drugs, production of monoclonal antibodies and research
of infectious diseases.
AWARDS AND DISTINTIONS
Annual awards of the Academy of
Sciences in Cuba in 2012
• Diana García del Barco et al. Demonstration of
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Laboratory of interaction analyses in Biacore
Responsible of the Laboratory
BSc. Vivian Huerta
[email protected]
phone: (537) 271 6022, ext. 3150
Biacore is an optical sensor which makes use of the
surface Plasmon resonance phenomenon (SPR) and
neuroportective effects of the coadministration of EGF
+ GHRP6 in biomodels of amyotrophic lateral sclerosis
and multiple sclerosis biomodels.
Isabel A. Guillen et al. In vitro and in vivo studies contributing
to discern the possible effects of Heberprot-P® in tumor
development.
Maria del Carmen Domínguez et al. Demonstration of
the therapeutic effect of two modified peptides derived
from the heat shock protein 60 kDa in experimental
models of rheumatoid arthritis.
Alicia Santos Savio et al. Identification of the first
peptide molecule that inhibits the biological effects of
interleukin 15.
Iris Valdés et al. Immunization strategy in monkeys
by heterologous prime-boost with virus Dengue-2 and
recombinant proteins containing domain III of the viral
envelope.
National Awards of Health 2013
• Diana Garcia del Barco et al. Preclinical studies of
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the treatment of neurodegenerative diseases with
EGF+GHRP6 as combined therapy. Award
Lázaro Gil González et al. Nucleocapdi-like particles
obtained obtained from the recombinant protein of the
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YEARBOOK 2013
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Dengue virus 2 capsid induce a functional and serotype-specific cellmediated immunity in mice. Award.
Iris Valdés, Alienys Izquierdo et al. The fusion protein domain IIIcapsid of Dengue virus 2 induces a functional and serotype-specific
immune response. Mention.
Enrique Iglesias Pérez et al. Two schedules for the mucosalparenteral coadministration of a multiantigenic formulation against
the HIV-1 in Balb/c mice. Mention.
Awards and recognitions
• Therapeutic vaccine against Chronic Hepatitis B. Second award at
•
•
the 21rst conference of the National Association for the Studies of
the Liver, India.
Giselle Pentón Rol et al. Latin American Award of Pharmacology
Dr. Plutarco Naranjo Vargas in memoriam (sponsored by the Latin
American Association of Pharmacology (ALF) and the International
Union of Basic and Clinical Pharmacology (IUPHAR). Neuroprotective
effect and molecular mechanisms of C-Phycocyanin in experimental
models of Multiple Sclerosis and Brain Ischemia.
Dr. Carlos Cabal was designated by the Université Rennes as
member of the Academic Board for a PhD degree in France.
Posters awarded in Latinfarma 2013
• Noraylis Lorenzo et al. Therapeutic effect of an altered peptide ligand
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derived from heatshock protein 60 by suppressing of inflammatory
cytokines secretion in two animal models of rheumatoid arthritis.
Alieski Cruz-Ramírez et al. Phycocyanobilin induces a protective
gene expression profile and restores the redox balance in a model of
acute cerebral hypoperfusion in wistar rats.
Marcelo Nazabal-Gálvez et al. C-Phycocyanin and interferon beta:
molecular mechanisms associated to a new combined therapy for
multiple sclerosis in the experimental autoimmune encephalomyelitis
model.
EVENTS AND COURSES
• Workshop on Bioinformatics and Biotechnological Applications, held
in HUSA Hotel Cayo Santa Maria, Junio 5-7, 2013.
• I Peptide International Symposium, Cayo Santa María, June 4th- 7th,
2013.
FOREING VISITORS
• Dr. Marie-Luise Michel, Pasteur Institute, France.
• Dr. Maryline Mancini, Pasteur Institute, France.
• Dr. Bernard Fanget, subdirector CMC, Wittycell, France.
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YEARBOOK 2013
• Dr. Eduard Escrich Escriche. Professor of Physiology at the Faculty
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of Medicine of the Autonomous University of Barcelona. Member of
the Board of Directors of Autonomous University of Barcelona.
Dr. Osvaldo Miguel Yantorno, National Director CABBIO-SECyT,
Memeber of the Directive Board CINDEFI (CONICET), Vice-president
of SAMIGE. National Autonomous University of La Plata, Argentina.
Dr. Rosana Mariel Romano, Vide-director of the Center for Inorganic
Chemistry of National University of La Plata, Argentina.
PUBLICATIONS
Forty-three scientific articles were published in international and national
scientific journals.
SUMMARY
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YEARBOOK 2013
Direction of Technological Development
T
he Direction of Technological Development (DTD) at the CIGB aims at converting
scientific results into final products and/or technologies. It also works on the continuous
improvement of the commercial products. The whole process of development comprises
the establishment of technologies at laboratories with all required quality parameters, until
scaling and pilot batch production; ruled by a methodology of integrated direction of projects,
which ensures proper planning of the objectives and a systematic review of its progress
In this direction, Good Manufacturing Practices (GMP) and Good Laboratory Practices
(GLP) are combined and sustained on a documented quality system certified by NC-ISO
9001:2008, providing the highest quality and safety of products and technologies that are
being developed.
The DDT is the organization dedicated to the design and development of biopharmaceuticals,
which aims to achieve a high scientific and technical level of high international standard in
various aspects of modern biotechnology. It has a highly qualified staff, characterized by its
adherence to moral and ethical values in the Cuban society.
Four departments that work coordinately constitute DTD: Fermentation Development
Department, Purification Development Department, Formulation and Packaging Department,
Documentation and Analytical Development Department.
DIRECTOR
Dr. Rolando Páez Meireles
[email protected]
Phone: (537) 250 4439
Key Executives
Head of the Fermentation Development Department
Dr. Jorge Valdés Hernández
[email protected]
Telephone: (53-7) 250 4448
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YEARBOOK 2013
Head of the Purification Development Departament
Dr. Gabriel Márquez Perera
[email protected]
Telephone: (53-7) 250 4438
Head of the Formulation and Package Development
Department
MSc. Fidel Raúl Castro Odio
[email protected]
Telephone: (53-7) 250 4433
Head of the Analytical Development
and Documentation Department
BSc. Ramira Dinorah Torres Idavoy
[email protected]
Telephone: (53-7) 250 4446
Main Results
• Production and release of 23 batches of active
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pharmaceutical ingredients (API) and 24 (PT) of the finished
product with positive results. This allowed the progress of
projects and the initiation or ongoing of clinical trials.
Inclusion of improvements in the production process
of IFA nucleocapsid antigen of the hepatitis B virus
(HBcAg), a process that resulted in more consistent
and higher quality of the final preparation. Also started
to transfer the purification step to the production plant,
including the production of Nasvac® vaccine batches.
It was made possible that The Directorate of Quality
Assurance at the CIGB issued the certification of
primary cells banks (PCB) and working (WCB) for the
production of HBcAg. Negative genetic construct vector
was developed; the release of cell bank laboratory (BCL)
of negative material was finished, and an increase was
achieved by 1.5 times the levels of expression of the
protein of interest to shaker scale.
The design space that justifies the operating conditions
of reverse phase chromatography (RP) in the preparative
process of obtaining new Heberprot-P® was studied and
concluded.
Approval and initiation of clinical trial phase II / III of
pegylated interferon (PEG-IFN) patent in Brazil issued
by ANVISA-Center for State Control of Drugs, Medical
Devices (CECMED) was achieved.
Technological Innovation Award and the Annual Health
Prize for "Development of PEG-Heberon® for the
treatment of hepatitis C “ was obtained in Cuba.
• Stability of pentavalent Heberpenta® of Ecuador was
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demonstrated. In addition, it was possible to continue
the execution of the joint development project of the
vaccine in Ecuador.
The expiring date of 48-kDa PEG reagent was extended
from 9 to 24 months.
The approval of adjuvant Quimi-Hib® vaccine was
achieved for 24 months in industrial batches.
A lyophilized formulation of CIGB-210 peptide was
developed.
An efficient analytical service was guaranteed. 3239
trials were conducted and 21 745 samples were
processed with less than 0.8 % of rejection and 1.4 %
reanalysis of samples
The stability report that declares the demonstration of
12 months stability at 4 ° C and 6 months at 25 °C for
the lyophilized formulation of CIGB-500 (3 batches) was
approved.
Stability was demonstrated in shelf life (5 ± 3 °C) of the
0.1 mg and 0.4 mg lyophilized presentations of antigen
CIGB-247, for 18 months
Broad spectrum of activity and selectivity of CIGB-378
was demonstrated on in vitro models of cancer: panel
of 60 lines of the U.S. National Cancer Institute and
pancreatic cancer subpanel
The effect of FCEhurec pellets was evaluated in an
experimental ulcerative colitis biomodel in rats.
The high productivity technology platform was introduced
and implemented with the use of monolithic arrays in
several bench-scale and productive processes
The purification of the CIGB 550-E7 protein, IFA of a
cancer vaccine against human papilloma virus was
established.
Was validated and transferred into the production
system techniques gel filtration and ion exchange by
high pressure liquid chromatography (HPLC) for the
control of PEG-Heberon®.
Was validated and transferred to the production system
of analytical techniques for the control of reactive PEG48 kDa (degree of activation and purity.
The expression of IFN gamma with kanamycin marker
in pTRIP and pT7 promoter variants, and the expression
of IFN gamma truncated at the same marker were
achieved.
The Guarantee of Quality was ratified in 2013 to the
Directorate of Technological Development
Satisfactory inspection results to maintain ISO
9001:2008 certification.
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YEARBOOK 2013
Awards
• Annual 2013 Health Award for Technological Innovation, Cuba:
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Development of PEG-Heberon® for the treatment of hepatitis C in
Cuba.
Development of a novel formulation for a suppository that contains
recombinant streptokinase in the treatment of acute hemorrhoid:
Péter Murányi Award . Saúde, Brazil 2014.
Relevant Award in the XVI Municipal Science and Technology Forum:
Quality Management System in the Development Stage in CIGB
certified by ISO 9001:2008
Award in the XVI Provincial Science and Technology Forum. Quality
Management System in the Development Stage in CIGB certified by
ISO 9001:2008
Discussed Thesis
PhD Thesis
• Fermentation Process for the obtaining of plasmid pIDKE2 in
Escherichia coli to be used in a vaccine candidate against hepatitis C
virus. Odalys Ruiz Hernández, July 10th, 2013
MASTER'S DEGREE Thesis
• Optimization of the microencapsulation of the growth factor in
microspheres of polylactic acid obtained by the method of double
emulsion and evaporation of the solvent. Elian Cruz Peñalver, May
28th, 2013
Thesis
• Purification of the CIGB-550-E7 Recombinant protein. Rayniel Acosta
Travieso. Tutor: Dr. Miladys Limonta.
• Establishment of an apyrogenic process for the antibody fragment
CIGB166A. Yinet Cartaya. Tutor: MSc. Jorge Sánchez.
• Development of a fermentation process in an a templa cultivation
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type for obtaining recombinant streptokinase: Marlen Herrera. Tutor:
Dr. Jorge Valdés.
Design of a chemically defined medium to obtain the HB core protein
in Escherichia coli: Claudia Granados. Tutor: MSc. Saily Martínez.
Publications
Five articles were published in international journals and 19 technical
reports were made.
SUMMARY
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YEARBOOK 2013
Direction of Production
W
ith the founding of the Center for Genetic Engineering and Biotechnology (CIGB) in
1986, and the need for a kind of a facility in which non-industrial scale production of
biotechnology products in Cuba could be established, a pilot plant was created, transformed
over time into an industrial facility of modern and functional design, in accordance with the
regulations and national and international standards.
The Direction of Production is now a large production complex of units of a high modern
technology, important production capabilities and highly trained personnel, which is
responsible for the obtainment of biotechnology products, with a constant desire to excel.
These units include more than 12 000 square meters and include all production facilities for
various products, some of them unique, supported and endorsed by international patents in
clinical trials in different countries
Production Management's primary mission is to provide the products and services required
for the domestic consumption and trading exports run by the commercializing company
Heber Biotec SA.
The vision of Production Management is geared to the strengthening of the product portfolio,
fostering innovation and working to meet the highest quality standards, taking into account
the growing demand for our products, the wide international recognition of the progress of
Cuban biotechnology industry, and its entry into the most demanding markets
It comprises the production units where the active pharmaceutical ingredients (APIs) are
produced to manufacture the anti-hepatitis B and Haemophilus influenzae tipo b vaccines,
the recombinant human epidermal growth factor (EGF), the granulocyte-colony stimulating
factor (G-CSF) the recombinant human interferons α and ϒ, pegylated interferon, the
transference factor, and monoclonal antibodies.
Along with them, are located the Department of Formulation and Packaging and those
supporting the activity of the direction: Services, Engineering, Process Control, Analysis
and Organization Management, Human Resources and Technology Transfer.
DIRECTOR
Ing. Jorge Luis Vegas Elías
[email protected]
Telephone: (537) 271 6022,
Ext. 2000
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YEARBOOK 2013
Key executives
Vice-director of production
Eng. José Enrique Brito León
[email protected]
Phone: (537) 250 4228
Head of the Regulatory Management Department
Bach. Luciano Francisco Hernández Marrero
[email protected]
Phone: (537) 250 4230
Vice-director of Operations
Eng. Manuel Enrique Montané Enríquez
[email protected]
Phone: (537) 250 4273
Head of the Hepatitis B Department
Eng. Juan Miguel Rivera Martin
[email protected]
Phone: (53-7) 250 4276
Head of the Multi-purpose Plant for Clinical Trials
MSc. Vivian Pujol García
[email protected]
Phone: (537) 271 6022, ext. 2114
Head of the Technology Transfer Department
Eng. Lilia Luisa Pérez Suárez
[email protected]
Phone: (537) 250 4274
Head of the Monoclonal Antibody Department
PhD. Rodolfo Valdés Veliz
[email protected]
Phone: (537) 271 6022, ext. 7101
Head of Department operatively working as a vicedirector of liophilized
Eng. Oscar Cruz Gutiérrez, MSc.
[email protected]
Phone: (537) 250 4464
Head of Granulocyte Colony Stimulating Factor
Department
MSc. Natacha Pérez Rodríguez
[email protected]
Telephone: (537) 250 4493
Head of the Transfer Factor Department
MSc. Yanieyis Alvarez Delgado
[email protected]
Phone: (537) 250 4533
Head of the Haemophilus influenzae type b Department
Eng. Belinda Díaz Montel
[email protected]
Phone: (537) 250 4270
Head of the Production Engineering Department
Eng. Iriac Bisquet Ramírez
[email protected]
Phone: (537) 250 4250
Head of the Process Control Department
PhD. Julio César Sánchez
[email protected]
Phone: (537) 271 6022, ext. 5951
Head of the Formulation and Packaging Department
Eng. Carmen Rosa Chuay Silva
[email protected]
Phone: (537) 250 4231
Head of the Human Resources Department
Eng. Mayra Ponce Guerrero, MSc.
[email protected]
Phone: (537) 250 4272
Head of Services to Production Department
BSc. Yutdelis Roben Aguilar
[email protected]
Phone: (537) 271 6022, ext 2005
Head of the Company Organizational Analysis Department
Eng. Francisco José Castañeda Márquez, MSc.
[email protected]
Phone: (537) 250 4232
RELEVANT RESULTS
Institutional achievements
• Effective improvements in the production process of
HBsAg API.
• Compliance of the training plan under the principle of
quality management.
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YEARBOOK 2013
• Fulfillment of the production plan of AcM CB.Hep-1 in
• Satisfactory results in the TT vac Hib to ChangHeber,
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• Signing contracts with Blood Banks, suppliers of the raw
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ascites and compliance of the inmmunogels export plan.
Efficient and uninterrupted analytical and microbiological
support to research-development productive groups, in
the center and also to centers that belong to (OSDE)
BioCubaFarma.
Consolidation of the work with the JVC-ChangHeber as
a strategy for productive output abroad.
Successful implementation of the production of
Heberprot-P® in Praxis, Vitoria, Spain.
Maintenance of the high operational reliability in the
auxiliary systems of the Production Plant. Improvements in the effectiveness of qualification and
validation work performed at the Engineering Production
Department.
Strategic improvements in the documentary system,
leading to greater efficiency and organization.
Successful maintenance activity in the productive area
by the Production Engineering. Department.
Efficient response to changes in water systems to meet
the production needs of the production plant in 2013.
Successful implementation of human capital actions in the
Production Management Direction as part of the assuring
operations for 2013. The results were satisfactory to
regulatory inspections, no increase in staff and a turnover
rate less than the 8 % planned and a higher rate of
mobility by 32 % of the workforce.
Relevant results in the service area with a significant
impact on the standard image of the Production Plant
and guaranteeing the transport operations.
Adequacy of the liquid waste tank, including the local
of synthetic peptides production, and implementation
of reactive donations or idle to other institutions, such
as alternative solutions in the Comprehensive Plan of
Waste Management of CIGB. Implementing solutions that allowed continuing treatment
of biosolids generated in the CIGB, during the capital
maintenance of the incinerator.
Completion of risks studies and vulnerabilities of
Peptides Plant (investment),Plant 4, Warehouses,
Plant 13 and Green Houses that belong to Agricultural
Research units, done by Inversiones Gamma.
Effective solution to the destruction by mechanical
grinding of finished products declared, due or noncompliant in the CIGB.
Satisfactory results in the execution of the Technological
Transfer Project and supplies of recombinant or human
IFN alpha 2b to BioM, Brazil.
China.
material for the production of API Transfer Factor. • Positive productive results were achieved during
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campaigns of IFA G-CSF, transfer factor and gamma
IFN.
Implementation of Plant 6 and plant 7 as service units
for products under development. Production of the API
of the hepatitis B virus capsid antigen (HBcAg) for its
use in Phase II clinical trials.
Report of the genotype Dam strain influence in the
mutation occurrence in the sequence of the recombinant
protein P64K after 56 generations. It was conducted
to address the regulatory body in Europe, for the
introduction of CimaVax-EGF vaccine in a clinical trial
in that region.
The Heberprot-P® delivery plan to Venezuela (250 000
bb 120 000 bb) was fulfilled for a total of 370 000 bb
exported for 144.3 million CUC. This was made possible
by the intense work to stimulate the consumption of the
product.
Product delivery was guaranteed by Heber Biotec SA to
fulfill the growing sales plan, both for exports and for the
domestic market, including deliveries to the Center of
Molecular Immunology (CIM).
API production of EGF in Plants 6 and 7 was implemented,
and the production plan of API Heberprot-P® was
fulfilled.
Certification by CECMED of the production systems on
Plant 1(HB) Plants 6 and 7 (IFN - PEG + parenteral EGF),
Plant 9 (FT), Plant 10, ChangHeber (IFN, IFN-PEG).
Maintain the quality system certification of CIGB
according to the ISO 9001:2008 standards.
Certification of the production of API Hib.Biocen by
CECMED.
Effective starting of Plant 4 with satisfactory results in
productive batches.
Satisfactory results in the production strategy for the
production of API in 2013 Quimi-Hib® in 2013 campaign
with temporary leave and the implementation of API
from Hib PRP supply of ChangHeber Results.
Fulfillment of the production plan of the activated PEG40
(367 g) ensuring PEGHeberon 2014 campaign and
works in ChangHeber.
Renovation of the licenses of Heberon Alpha R® liquid
3 M, 5 M and 10 M, Heberon Alpha R® lyophilized 3 M, 5 M
and 10 M, Heberon Gamma®, HeberPAG® 3 M and 10 M,
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YEARBOOK 2013
•
•
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Heberkinasa® 750 000 1 500 000 IU and IU , HeberNem®-L
and Hebertrans® in Cuba.
The condition of Relevant Center in developing the work
of the National Forum on Science and Technology has
been maintained for 8 consecutive years.
Validation of Lowry method for the quantification of total
proteins in samples of the synthetic vaccine against
Haemophilus influenzae type b production.
Ellman’s method validation for determining the
concentration of sulfhydryl groups to produce samples
of the synthetic vaccine against Haemophilus influenzae
type b.
Functioning of the activated carbon filter system of the
purified water in the production Plant.
Technical and scientific
achievements
• Production process of the Pegheberon active ingredient.
•
•
•
•
•
Threefold increase of the Active Pharmaceutical
Ingredient.
Correlation between LAL and rabbit Pyrogen test in
recombinant Human Epidermal Growth Factor samples.
Replacement of serum-supplemented medium in
the freezing CB.Hep-1 hybridoma and CB.Hep-1.
monoclonal antibody production.
Demonstration of the chromatography affinity stability
used to purify a monoclonal antibody as immunoreactive
in the manufacture of a vaccine against hepatitis B.
Detection of an immunologically important epitope in the
active pharmaceutical ingredient of the Heberbiovac HB®
vaccine employing CB.Hep-4 monoclonal antibody.
Assessment of two transgenic tobacco plant varieties for
the HBsAg specific plantibody production.
COURSES DELIVERED
• Requalification in Good Engineering Practices.
• Course on Inmovilization and Purification.
• Requalification in Good Cleaning and Sanitation
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•
•
•
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Practices.
Requalification in Good Documentation Practices.
Introductory Production Course.
Basic Production Course.
Seminar on the updated requirements of Good Practices
by Cecmed.
Course on Hygiene and Safety Requalification.
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YEARBOOK 2013
Visitors
• Officials from the Province People’s Power, Cuba.
• Dr. Miguel Díaz-Canel Bermúdez, First Vice-President
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•
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of the Councils of State and Ministers, Cuba.
Executives from OSDE BioCubaFarma (several visits).
Executives from the Fire Management Agency, Cuba.
Executives from Praxis Pharmaceutical, Spain.
Thirty students from the Architecture Faculty, University
of Havana.
Vice-Minister of Finance and prices, Cuba.
New graduated university students from OSDE
BioCubaFarma, Cuba.
Cecmed (trámites regulatorios, en varias ocasiones).
CECMED (regulatory affairs).
Ambassador and Commercial Advisor from the French
Embassy in Cuba.
Executives from WITTYCELL, France.
Dr. Fidel Castro Díaz-Balart, Scientific advisor for Cuba’s
Council of States.
Publications
There were approved to publish 13 articles and 37 technical
reports.
SUMMARY
51
YEARBOOK 2013
Biotherium
T
he Biotherium is the responsible for the use and care of lab animals and the control of
biotechnological products, as well as; the design and development of the training in
laboratory animal science at the Center for Genetic engineering and Biotechnology (CIGB)
in Cuba and the countries on request.
It counts with high-tech equipment including a rigorous automated control of environmental
parameters including: temperature measurements, relative humidity, and differential
pressure. Ammoniac concentration is performed using qualified digital equipment.
In this direction works scientists and highly qualified specialists with over 12 years experience
trained in different fields of biological science, quality management system and laboratory
animal science. Its main topics of research are the obtainment of biomodels assessing the
obtained products at the CIGB, experimental toxicology and pharmacology, histopathology,
environmental safety, bioethics, immunology and animal breeding.
Manager
Dra. Karelia Cosme Díaz
Phone: 250 4301
[email protected]
Mission
The Biotherium’s mission is to satisfy the demand of in vivo and preclinical studies to the
different directions of the CIGB, to guarantee not only annual requests not less than 98 %
to the Quality Control and Production Directions, but also 95 % annual requests to the
Research and Development Direction.
Furthermore, the direction designs and accomplishes not less 95 % requests of theorical
and practical training being requested throughout the year.
Main executives
Head of the Department of Animal Experimentation and Good Preclinical Practices
MSc. Jorge Castro Velazco
phone: 250 4347
[email protected]
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YEARBOOK 2013
Biotherium
Head of the Department of product control and materials
Omar Mosqueda Lobaina
Phone: 250 4306
[email protected]
Senior Specialist
Dr. C. Dania Bacardí Fernández. PhD.
Phone: 205 4342
[email protected]
Head of the Group of Experiments with laboratory
rodents
Laritza Gorovaya
Phone: 250 4306
[email protected]
Group of Good Preclinical Practices
MSc. Lizet Aldana Velazco
Phone: 250 4306
[email protected]
Main Results
• Obtainment of the Sanitary register, authorization
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of clinical trials (CT) or Renovation and trámites of:
Heberprot-P en Cecmed the files were presented in
Chemo, Costa Rica, Marruecos, Peru, Russia, Praxis,
Turkey and Bolivia. Report of approval by CECMED of
the final Report of CT Praxis, Heberpenta L: register
in Peru, Venezuela, Uruguay and Dominican Republic,
Peg-Heberon register request in Thailand, China and
Byelorussia, Proctokinase®: Approval SAEC/Cecmed,
Nasvac: deliver IMPD for Witycell and CRISTALIA
Enterprise, CIGB-247, CIGB-247A, CIGB-370, CIGB552, CIGB-845A: Approval by the CECMED of CT,
HeberPAG: development of the project Family.
Development of the completed toxicological qualifications
of impurities of Heberprot-P®, CIGB-500, CIGB-552,
CIGB-247A, CIGB-845a and the vaccine candidate
(Nasvac).
Re-certification of ISO 9000:2008, for 4 consecutive
years.
A researcher got a doctoral degree in Sciences..
The Guidelines for the use of Laboratory Animals and a
Conference in the Master's Degree of Trends in Modern
Biotechnology were delivered.
Development and validation of the method Determination
of the Pharmacokinetics and biodistribution in vivo of
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biomolecules marked magnetically from Images of
Magnetic Resonance in animal models.
First studies of FC/FD of CIGB-128-A in humans and
non- human primates. Final reports of the studies of FC/
FD of CIGB-128-A in primates and humans.
Preclinical Safety of CIGB-210 in repeated doses.
Improvement of the image in areas of the CIGB..
Effective regulatory activity on Biological, Chemical and
Radiological Safety.
Achievement of 14 Toxicological studies including the
Certification by Quality Assurance at the Cecmed and
the approval of SAEC.
Satisfactory results in the process of evaluation at work
achieving 95 % to the compliance of the process at the
center and 99 % in the production facilities
Quality Management of the Biotherium at the CIGB,
applied to the control of biotechnological products and
preclinical research.
Immunogenicity and safety of the synthetic vaccine
Quimi-Hib® and of the pentavalent vaccine Trivac HB® +
Quimi-Hib® concentrated.
Elaboration of the preclinical strategy of the GCSF-PEG.
Elaboration of the preclinical strategy of the vaccine
candidate Nasvac for the First World.
Obtainment of the ulcerative colitis biomodels.
Awards or distinctions
• Annual Award of the Academy of Sciences in 2012:
Proctokinase®, new medicine for the treatment of
hemorrhoids disease. Certification of 14 toxicological
studies and approval of clinical trials of assessed
products by the Quality Assurance Direction.
Internacional Projects
• Participation in different projects like: CIGB-300, CIGB247, CIGB-166a and Heberprot-P®.
Courses and seminars
Courses
• Guidelines on the care and use of laboratory animals in
researches.
• Requalification Guidelines on the care and use of
laboratory animals in researches.
• Good laboratory practices.
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YEARBOOK 2013
Biotherium
Events
• Congress AETOX 2013. Proof of the safety requalification
of the HTP.
Foreign Visitors
• Senior Specialists from the biological Laboratory in
Argentina.
• Senior Specialists from Praxis – Apointech.
Publications
Two articles and four technical reports were published.
SUMMARY
54
YEARBOOK 2013
Direction of Agricultural Research
T
he Direction of Agricultural Research from the Center for Genetic Engineering and
Biotechnology (CIGB) leads the Division of Plants and Animal Biotechnology. The first
involves conducting projects in Plant Molecular Virology, Functional Genomics, PlantMicroorganisms interactions, Plants as Bioreactors, Environmental Biotechnology, and
Plant Experimental Parcel. The second, aimed at projects related to Animal Health, Aquatic
Biotechnology, Expression in Mammalian Cells and Clinical Trials.
The CIGBs in Camagüey and Sancti Spíritus are integral part of the one in Havana, and
also develop major projects of agricultural profile.
The use of genetic engineering as a working method involves a responsibility for human
security and environmental protection. This is an essential method for the development of
new products and in the solution of problems of science and technology. Our mission is
aimed at such goals, based on ethical principles, which are performed by evaluating the
risks and following the most stringent biosafety measures as well as the requirements of
current legislation.
DEPUTY DIRECTOR
Dr. Carlos G. Borroto
[email protected]
Phone: 250 4201, 271 6032
Fax: 273 1779, 273 6008
Key Executives
Head of the Plant Department
MSc. Merardo Pujol
[email protected]
Phone: (5-37) 250 4182
Fax: (53-7) 273 1779
Head of the Department of Animal Biotechnology
Dr. Mario Pablo Estrada García
[email protected]
Phone: (53-7) 250 4423
Fax: 537 273 1779
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YEARBOOK 2013
Head of the Project of Plants as Biorreactors
MSc. Abel Hernández Velásquez
[email protected]
Phone: (53-7) 250 4371
Head of the Molecular Virology Project in Plants
Dr. C. Alejandro Fuentes Martínez
[email protected]
Phone: (53-7) 250 4369
Head of the Functional Genomics Project
Dr. Orlando Borrás Hidalgo
[email protected]
Phone: 271 6022, ext. 3151
Head of Plant-Microorganisms Interactions Section
Dr.C. Lázaro Hernández García
lá[email protected]
Phone: 271 6022, ext. 3116
Head of the Environmental Biotechnology Project
Dr. Camilo Ayra Pardo
[email protected]
Phone: (53-7) 250 4370
Research is also being conducted on genetically modifying
crops to improve their properties, for their subsequent
use in industry and in animal feeding. Such crops involve
sugar cane, potato, tomato, rice, sweet potato, maize and
soybeans. Working in citrus, fruits, and bananas is also
included in the study in collaboration with Cuban scientific
institutions. These research works are based on the strictest
compliance with biosafety regulations in force, and promote
the study of toxicological and environmental implications of
genetically modified organisms.
The expression of pharmaceutical proteins in transgenic
plants is a goal of special interest in which laboratories of partner
centers in Sancti Spiritus and Camagüey are also involved.
The identification of genes associated with resistance to
disease and environmental adversity (salinity and drought)
are pretended to be sought with the genomic studies of
these crops.
RELEVANT RESULTS
Corn genetic improvement
• The S2 generation from maize line MIR162, progenitor
Head of the Soybean Biotechnology Section
Dr. C. Gil Enríquez Obregón
gil.enrí[email protected]
Phone: (53-7) 250 4355
Head of the Veterinary Clinical Trials Section
Bsc. Marisela Suárez Pedroso
marisela.suá[email protected]
Phone: (53-7) 250 4420
Head of the Animal Health Section
Bsc. Alina Rodriguez Mallon
[email protected]
Phone: (53-7) 250 4407
PLANT DIVISION
The Division of Plants is devoted to molecular biology and
the genetic improvement of target plant species as well as
the research of compounds and microorganisms useful for
crop protection, food, human and animal health.
Ongoing projects are aimed at the introduction of genes for
plant defense against pests and fungal and viral diseases.
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of simple hybrids with high genetic value was obtained.
It contains the new Vip3A specific insecticidal protein
against corn moth. This protein has a mechanism of
action against larvae of the insect pest that is different
from the Cry1Fa protein so it can be used for the
management of insect resistance.
Three hybridomas and their respective seed banks,
which produce monoclonal antibodies against the
insecticidal protein Vip3A were obtained.
An ELISA system was established for the detection of
the Vip3A protein in plant extracts.
It was demonstrated that the maize moths show a strong
preference for ovipositing in Bt maize plants when
damage to conventional maize is high. The implications
for the evolution of insect resistance were studied.
Insect proteins were expressed in Pichia pastoris to
begin studies of its utility as targets for insecticides.
Soybean genetic improvement
• Transgenic soy clones from DT84 and IncaSoy-36
varieties were obtained. The genes of defensin and
resistance were expressed showing the resistance to
Asian rust and other fungi of importance in the soybeans
harvest.
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YEARBOOK 2013
• New lines derived from a cross between parental
•
soybeans with higher productive potential, reaching the
F4-F5 generation were achieved. Progress was made on
the basis of molecular techniques in the characterization
of soybean genotypes relevant to the breeding program,
in addition to genes associated to contrasting phenotypes.
Scaling of transgenic soybeans in 20 acres in the
Military Agricultural Enterprise Cubasoy was achieved
demonstrating the productive potentials of the lines obtained.
The expression of more than 10 g of anti-HBsAg per kg of
snuff seeds and 0.18 g/kg of leaves was achieved.
Control of diseases caused by germinivirus
• The study of the silencing RNA geminiviral silencing
Soybean biofertilizer production
• Production plan and schedule of sales (1000 L) of
•
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Bradyrhizobium inoculant was fulfilled. Soybeans
planted in the UAM Cubasoy and areas for extension of
glyphosate-resistant transgenic variety in different parts
of the country was inoculated.
The average viability of the inoculum was 3 x 1010 c.f.u./
mL. The inoculated plants showed effective nodulation
and good nutritional status. Urea application was saved.
Writing of the technological package for the manufacture
process of Bradyrhizobium inoculant for soybeans was
conducted.
Biocatalysts
• Design and development of biocatalysts for enzymatic
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conversion of sugar cane products with high added value,
such as fructooligosaccharides (FOS) and invert syrup.
The international presentation of a patent which claims
the production in P. pastoris of a plant fructosyltransferase
(1-SSTrec) and its use in conversion of sucrose to
1-kestose was performed.
It was determined that 1-SSTrec secreted in P. pastoris
is a glycoprotein. Mannose chains play an important role
in the catalytic activity of the enzyme.
It was also evidenced that 1-SSTrec is useful to increase
the content of fructans in chicory root extract, a new
commercial utility of the enzyme.
A solid formulation of thermostable invertase method was
achieved by drying at 60 °C without loss of enzymatic
activity, and extremely stable in non-refrigerated storage.
Plants as expression systems
In the Plants as Bioreactors project we continued to
develop strategies that increase the expression levels of
pharmaceutical molecules.
•
process in species of plants of economic importance
could point to new approaches for obtaining resistance
against the most represented and disseminated viruses
in the world. In this sense several models of plant-virus
interaction using TYLCV and ToMoTV species were
established. Different patterns of behavior were defined
and its effect on plant transcriber is analized.
The obtaining of molecular tools derived from viral
genomes is another objective of the present project.
The design of replicons of geminiviral origin capable
of expressing or silence specific genes is an effective
tool for functional genomics and the use of plants as
bioreactors. Several designs of this type of vector were
obtained from the genome of ToMoTV, as the integrative
and systemic expression of a specific gene.
Plant genomics
Identification, isolation and characterization of new genes
involved in disease resistance. The worldwide production
of economically important crops has been considerably
affected by the incidence of various types of pathogens. The
project is based on the study of the major molecular and
biochemical mechanisms of disease resistance in plants,
with the aim of identifying new genes in the plant involved
in recognition, signaling and plant response to diseases.
The project uses tools like SuperSAGE combined with next
generation sequencing to identify the genes. In addition to
using different strategies for the analysis function by RNAi.
It was identified a new receptor immune response in plants,
and a strong candidate for biological activity of the plant
defense molecules was achieved.
ANIMAL BIOTECHNOLOGY DIVISION
The Division of Animal Biotechnology is composed by three
main research groups.
The first, Animal Health, developed a vaccine against the
tick Boophilus microplus tick, known as Gavac®, with a large
and very successful use in Cuba, with very important field
experiences in other countries. New antigens against ticks
are currently developed, which increase the efficacy of the
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YEARBOOK 2013
existing vaccine. They are also working on the development
of new veterinary subunit vaccines, genetically engineered
diseases that affect economic species, as well as the
design and production of diagnostic systems against such
veterinary diseases.
The Aquatic Biotechnology group has researched about
nutrition and stimulation of growth and the immune system
of aquatic organisms for more than fifteen years. It has
achieved a remarkable growth and stimulation of the immune
system of shrimp and fish by adding nutritional supplements
(metabolic modifiers). Acuabio 1® is the first commercial
product of this group, recorded with very encouraging results
in field tests in Cuba and other countries. Currently has been
isolated and studied first neuropeptides and antimicrobial
peptides, with a very important role in the immune system of
fish function. It has been possible to study the activation and
modulation of immune response to both innate and acquired
in response to the biological activity of the isolated peptides.
These molecules could provide new biotechnological tools
for application in modern aquaculture and animal health.
The Clinical Trials Group is dedicated to preclinical field
study of vaccines and veterinary drugs that are generated
in the department as well as the establishment of rapid
diagnostic system to support the implementation of plans
for the eradication of diseases and an accurate diagnosis
of important pathogens that threaten the main livestock
produced worldwide.
The Department also generates model transgenic animals,
being established pro-nuclear injection technology using
lentivirus system with a high efficiency. They have managed
to develop models in mice, rabbits, cows.
Project of new antigens against ticks
It has been possible to demonstrate the efficacy of a
peptide of the ribosomal protein P0 against Rhipicephalus
sanguineus ticks and Rhipicephalus (Boophilus) microplus.
At present various immunostimulatory molecules to ensure
adequate response against the peptide and its production is
feasible from a technological and economic perspective are
discussed. Besides, the effectiveness of the new candidate
against strains of different regions worldwide and different
species of ticks is studied.
Aquatic Biotechnology project
It has been possible to demonstrate the activation and
immune response modulation of both innate and acquired
in response to the biological activity of the isolated peptides
(e.g. PACAP). Three novel peptides have shown a great
activity as antimicrobial agents in a wide spectrum of Gramnegative bacteria, Gram positive bacteria and fungi. Studies
have been carried out by qPCR of differential expression in
different tissues isolated from fish, in normal conditions and
after infestation with bacteria and viruses. The Litopeneus
vannamei shrimp PACAP peptide was first isolated being a
pioneer discovery in the characterization of these peptides
in crustaceans. There is a vaccine candidate against sea
lice (sea lice) that attacks salmonids in both north and south
seas, with trials with promising results, that could provide a
great tool for the control of these ectoparasites that cause
millions in losses.
National use of Gavac® in Venezuela as part of a
comprehensive program of tick control
• A 60 % compliance of the reimmunization plan for
•
2013 and 74 % of new additions to PCIG in Venezuela
was achieved (XII-XIII Joint Commission from 2012 to
2013) The PCIG project started in Venezuela, XIII Joint
Commission 2013.
There were 1 907 054 animals included in the Integrated
Program forcattle tick control. It has been a record in
the history of the world. 28 332 farmers have benefited
and 5475 have been trained. The results obtained show
a 81.6 % reduction in costs and a reduction in cost
per animal for tick control of 86.65 % representing an
estimated benefit to date, only in respect of chemicals
78 737 300.00 BsF ($ 12 497 984) which is 260 tons
less of chemical released into the environment.
Antibody testing conducted on a sample confirmed the
response to immunization and damage was observed
in the reproductive potential of ticks in the field, so the
expecting results are fulfilled and the objectives are met.
An intense work of colleagues in complex conditions,
as well as the entire production, export control and the
immunogenic insurance has been required.
Strengthening of the Integrated Control Program
of bovine tick in Cuba
• Gavac® promoters were located by provinces across the
country, since October 2013.
• Product purchases for producers through contracts with
Labiofam are established.
promotion (folding,
• Programme
television
and
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YEARBOOK 2013
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newspaper reports) encouraging producers about the
benefits of this technology.
The work of IMV-CIGB – ANAP has strengthened to
achieve the results of science in the country.
There were 515 101 animals included in the program, of
which 233 990 in the private sector and 281 111 of the
state sector, and 157 070 were new inclusions
We have trained 3246 people in 2013 on the bases of
PCIG.
The work guaranteed Gavac® record sales this year.
The necessary foundations for reintroduction and
national expansion were created and an economic impact
is achieved in the animal, human and environmental
health, considering that there is only one health.
Development of new vaccine candidate against
E2CD CSF, produced in mammalian cells in
suspension
Assay No. 5 was done. In it the quality of the E2CD antigen
is reaffirmed of obtaining a high antibody response which is
then multiplied by the 2nd booster dose. Vaccination with
the candidate reversed the immunosuppressive effect. The
action of the Montanide ISAV2 adjuvant is valued as the
protective element. The ability to achieve viral immunogen
sterility 7 days after the strain confrontation. is shown .The
high titers of neutralizing antibodies eliminate the virus
before it can replicate systemically in the animal.
Working cell banks and new virus seed banks will be created,
once NPLA procedure conditions are reviewed and adjusted.
Advances in the development and assembly
of NPLA techniques to measure antibodies
against CSF
• The documentation was established according to GLP
and Biosafety for the work in different areas.
Good Laboratory Practice
Certification of Good Laboratory Practice in the field of
Agricultural Research by Quality Assurance division at the
CIGB was obtained.
AWARDS
ANNUAL AWARDS OF THE ACADEMY OF SCIENCES OF
CUBA - 2012
• Antimicrobial peptides as molecular therapeutic agents
•
•
• The technique for the evaluation of neutralizing
antibodies of the PPC virus was standardized by direct
immunoperoxidase method, which has simplified the
work and time. Results are easily interpreted similarly
by several analysts. Primary and working banks for used
cells and viruses were created by providing a method of
fixing the cells for the direct revealed at CIGB multiple
anti E2 peroxidase labeled conjugates. This technique
is only performed at CENSA, to have her in our center
allows us better control of the animals included in the
trials and directly know the status of swine mass also
allows us to maintain a serum panel.
•
and molecular adjuvants for fish and mammals. Jannel
Acosta1, Mario P Estrada1, Yamila Carpio1, Vivian Montero2,
Janet Velázquez1, Iris Valdés3, Reynold Morales1, Antonio
Morales1, Yasser Zamora1, Hilda Elisa Garay4, Osvaldo
Reyes4, Elsa Rodríguez1, Sonia González5, Ania Cabrales4,
Yordanka Masforrol4. 1 CIGB, División de Biotecnología
Animal. 2 Cidem, Departamento de Bioquímica. 3 CIGB,
División de Vacunas. 4 CIGB, División de Química-Física. 5
CIGB, Departamento de Patentes.
Inhibition of molecular patterns associated to pathogen
confers high protection against fungi and oomycetes in
plants. Ingrid Hernandez Estévez1, Roxana Portieles
Alvarez1, Yussuan Silva2, Merardo Pujol Ferrer1,
Orlando Borrás-Hidalgo1, Osvaldo Oliva Borbón1. 1
CIGB, División de Plantas. 2 Instituto de Investigaciones
del Tabaco, Cuba.
Development of biocatalysts based on thermostable
Thermotoga maritima invertase for sugar cane total
hydrolysis. Carmen Menéndez1, Duniesky Martínez2,
Lázaro Hernández1, Enrique Pérez2 Luis E. Trujillo1,
Yuliet Mazola1, Bessy Cutiño-Avila3, Alberto del MonteMartínez3, Ernesto González1, Alina Sobrino Legón2,
Ricardo Ramírez1, Merardo Pujol1, Carlos Borroto1. 1
CIGB, Dirección de Investigaciones Agropecuarias
2
CIGB de Sancti Spíritus. 3 Centro de Estudios de
proteínas, Facultad de Biología, Universidad de La
Habana (UH).
Determination of the mechanisms of Tsukamurella
paurometabola C-924 plant growth promotion. Marieta
Marín1 Jesus Mena1, Idania Wong1, Rolando Morán1,
Ramón Franco1, Pavel Chavelis2, Graciela García3,
Rosa Basulto1, Armando Hernández4, Liuven Veloz,
Eulogio Pimentel1 y Alain Moreira1. 1 Centro de Ingeniería
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YEARBOOK 2013
Genética y Biotecnología. 2 Estación Experimental de
Suelos. 3 Laboratorio Provincial de Sanidad Vegetal. 4
Centro Nacional de Investigaciones Científicas, Cuba.
Forum on Science and Technology 2013
• Results of the Program Integrated Control Program
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against Cattle Tick with the immunogen Gavac® in Cuba
and Venezuela. Relevant Award at municipal levels.
Mention at Havana Province. Prominent, CIGB.
Implementation of Good Laboratory Practice in the
Department of Agricultural Research. Prominent, CIGB.
Standardization at the CIGB of the technique of
neutralization of classical swine fever (CSF) by specific
antibodies labeled with peroxidase (NPLA) virus.
Mention, CIGB.
Scientific and Technical Awards 2013
• 1-SSTrec, enzyme without similars in the market for
•
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•
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• Poster: Tobacco seeds as an alternative platform to
•
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fructosidase thermostable
•
• First report of the Pituitary adenylate cyclase activating
•
EVENTS
commercial production of fructooligosaccharides (FOS).
• Immobilization for rational design of an exo-β-
•
Rhipicephalus microplus tick using Heberbiogar® in
Venezuela.
peptide of the (PACAP) in crustaceans: conservation
of its biological function as a growth promoter and
immunomodulator in the white shrimp Litopenaeus
vannamei.
New developments in the characterization of genes /
proteins in akirins lice sea.
Inhibition of the transmission of pathogens between
ticks by vaccination with Bm86.
Lactobacillus pentosus IC1, an isolation of fermented
vegetables with probiotic and immunostimulating activity
in shrimp
Development of immunochemical tools for the detection
of transgenic corn event MIR162 resistant to Spodoptera
frugiperda.
Effective Beta-lactam antibiotics for removing
Agrobacterium tumefaciens in calluses from Cuban rice
varieties.
1-SSTrec, enzyme without similar for the commercial
production of fructooligosaccharides (FOS).
INTERNATIONAL PROJECTS
• Project XII and XIII CONCUVEN Joint Commission.
Implementation of a program for the control of
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production of pharmaceutics. Hernández Abel, López
Alina, Ceballo Yanaysi; Rosabal Yamilka, Cabrera
Gleysin, Simón Margarita, Pérez Marlene, Ramos
Osmany, Tiel Kenia, Rodríguez Meilyn, Enríquez Gil.
Poster: Antitumor activity of the aglycosylated antibody
against human epidermal growth factor receptor
(EGF-R) produced in transgenic plants. Rodríguez M,
Pérez L, Gavilondo JV, Bequet-Romero M, Garrido G,
Huerta V, González EM, Cabrera G, Soto J, Perez M,
Ramos O, Sánchez B, Ayala M, Triguero A, Cremata J,
Álvarez A, Morera Y, Sánchez J, Lamdán H, Oliva O,
Simón M, Pujol M, Hernández A.
Physiology and plant biotechnology and its applications
in improving the environmental stress. UH. 2012.
Fructan metabolism in endophytic diazotrophs of the
genus Gluconacetobacter (lecture).
Catalytic properties of N-glycosylated Thermotoga
maritima β-fructosidase overexpressed in Pichia
pastoris (poster).
Advances in the regulatory status of GM soybean in Cuba:
Extension in different production areas. Gil A Enriquez,
Natacha Soto, Yamilka Rosabal, Celia Delgado, Aneisi
Reyes, Aleines Ferreira, Odet Céspedes, Raul Armas,
Orlando Borras, Roxana Portieles, Osvaldo Oliva,
Merardo Pujol, Carlos Borroto.
Lecture: Biotechnology platform in bacteria, yeast and
mammalian cells to produce veterinary subunit vaccine.
Production of highly polymerized bacterial levan in
transgenic tobacco plants (lecture).
Genetic transformation of Cuban soybean variety
Incasoy-36 mediated by particle bombardment of
embryonic axis. Natacha Soto, Celia Delgado, Yamilka
Rosabal, Aleines Ferreira, Gil A Enríquez.
Evaluation of salt tolerance in 10 varieties of soybean
(Glycine max (L.) Merryl. Yuniet Hernández, Marilyn
Florido, Natacha Soto, Celia Delgado, Rodobaldo Ortiz,
Gil Enriquez.
Obtaining of soybean lines resistant to glyphosate
herbicide soybean breeding by artificial crossing. Celia
Delgado, Gil A Enríquez, Rodovaldo Ortiz, Natacha
Soto, Aneisi Reyes, Odett Cespedes, Eliudimir Peña,
Moisés Morejón.
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• Obtainment of soybean plants carrying epsps and
•
•
•
•
•
•
•
•
•
•
•
•
•
defensin (NmDef02) genes. Natacha Soto, Celia
Delgado, Yamilka Rosabal, Aleines Ferreira, Orlando
Borrás, Roxana Portieles, Aneisi Reyes, Yuniet
Hernández, Gil A Enríquez.
Cloning and functional characterization of three novel
antimicrobial peptides from tilapia (Oreochromis
niloticus).
A novel GH secretagogue, A233, exhibits enhanced
growth activity and innate immune system stimulation
in teleosts fish.
Functional and evolutionary analysis of the fructan
metabolic in endophytic bacteria of the genus
Gluconacetobacter (conferencia).
Analysis of response of two silencing events immune
to TYLCV in different tomato genotypes under extreme
conditions.
Poster: Inhibition of Erlichia canis and Babesia canis
transmission in dogs vaccinated with Bm86 antigen
(Gavac®) Rodriguez Mallon A, Zacarias RM, Benavides
Ortiz E, Gómez Ramírez AP, Javier Andres JO, Soto
Rivas JL, Sampaio PH, Evora PM, Bechara GH.
Implementation and extension of an Integrated Control
Program against bovine tick with the immunogen Gavac®
(April-May 2012 and December 2012 - January 2013).
Prebiotics and Probiotics (Professor, pre-congress course).
Training Workshop: Implementation and extension of an
Integrated Control Program against cattle tick with the
immunogen Gavac® (December 2012 - January 2013
and June-July 2013).
Workshop: “News and perspectives of a byproduct to
control ticks "(May 14-15). CIGB Camagüey. Danny
Pérez, Mario P Estrada, Pedro Encinosa.
Lecture: Culture of vitro cells. Its importance in the
development and regulatory compliance in biological
products. María Pilar Rodríguez.
Course on Virology. Master's Degree of Trends in
Modern Biotechnology. Lecturer: María Pilar Rodríguez.
2013-Jan 2014.
2013, IX Latin American Symposium on alcohol and
yeast production, Varadero, (April 08-11). Poster:
Sucrose hydrolysis by a bacterial β-thermoestable
highly exofructosidase produced in the yeast Pichia
pastoris. Carmen Menéndez, Duniesky Martínez, Luis
E Trujillo, Enrique R Pérez, Ricardo Ramírez, Alina
Sobrino, Lázaro Hernández.
International Workshop on Biosafety 2013 . September
16-19. Habana Libre Hotel, Cuba. Biosafety assessment
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•
•
for recombinant protein production in the mammary gland
of goats by the use of adenoviral vectors: preliminary
study.
Biosafety Regulatory documentation for transfer and
manipulation of classical swine fever virus strains in the
CIGB.
International Congress ALPA2013 . XIII Meeting of the
Latin American Association of Animal Production. IV
Congress of Tropical Animal Production. November
18 to 22. Conventions Palace, Cuba. Advances of the
Integrated Control Program against Cattle Tick against
with Gavac® immunogen in Cuba.
AWARDS
XVI Forum on Science and Technology
• Results of the Integrated Control Program against cattle
•
•
•
tick with the immunogen Gavac in Cuba and Venezuela.
Relevant.
Implementation of Good Laboratory Practice in the
areas of IAP.
Standardization of the neutralization technique to
classical swine fever (CSF) by specific antibodies
labeled with peroxidase (NPLA). mention in the XVI
Forum on Science and Technique. Municipal Level.
Playa, La Habana.
Results of the Integrated Control Program against Cattle
Tick with Gavac® immunogen in Cuba and Venezuela.
Relevant.
PUBLICATIONS
Eight articles were published in international journals.
SUMMARY
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DEPARTMENT OF INFORMATICS AND COMMUNICATIONS
T
he Department of Informatics and Communications is in charge of everything related to
computing resources and use, software development, process automation as well as
communications, both telephone and radio. It is integrated by 42 workers, 22 professionals
and 13 technicians.
The direction is divided into seven areas as follows:
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Direction: 2 members.
Networks: 7 members.
Bioinformatics: 10 members.
Development of Applications: 13 members.
Process Automation: 5 members.
Technical Assistance: 5 members.
Communications: 5 members
DIRECTOR
Dr. Ricardo Ricardo Parellada
[email protected]
Phone: (537) 250 4156
Key Executives
Head of the Development of Applications Group
Ing. Julio César Iglesias Bayeux
[email protected]
Phone: (53-7) 250 4157
Head of the Network Group
Ing. Ronny Córdova Díaz
[email protected]
Phone: (53-7) 250-4157
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Head of the Group of Automation of Processes
MSc. Noel Jesús Reyes Fleitas
[email protected]
Phone: (53-7) 250-4271
Head of the Communications Group
Téc. Amaurys Cardoso Álvarez
[email protected]
Phone: (53-7) 250 4211
Head of the Technical Assistance Group
Ing. Wilfredo Alemán Fernández
[email protected]
Phone: (53-7) 250 4210
MAIN RESULTS
• Desvelop new Products and market based on a Strategic
Plan 2012-2016.
• Complete the research stage (Biomedical and agriculture).
Priority projects at the CIGB
Heberprot-P®
• Data Analysis of results by quantitative (qPCR) of 49
genes and search for response-associated patterns.
CIGB-300
• Analysis of the differential expression of genes and
•
discussion of results of two experiments by qPCR in this
project.
Identification of substrates of casein kinase 2 (CK2)
linked with biological routes and illnesses of interest.
CIGB-378
HeberProvac
• Presentation in the Commission of Bioinformatics of a
summary of the work made with a proposal of possible
biomarkers.
Tomato
Results of the bioinformatic analysis of the experiment of
small size RNA deep sequencing in samples of transgenic,
non-transgenic tomato, exposed and not exposed to the
environment.
• Identification of sequences of geminivirus known from
the analysis of RNA samples of small size.
• Identification of the partial sequence a possible new
virus from the analysis of small size RNA samples.
• Identification of potential target of RNA of interference
derived from the speckled yellow virus of the Cuban
tomato leave(TYLCV) in the genome of the tomato.
• Identification of potential target of RNA of interference
derived from the transgene in the genome of the
tomato and in the geminivirus with genome completely
sequenced.
• Identification of the coincidences between the genes
differently expressed in the study of microarrays to
evaluate the off-targets effects of the transgene and the
transcripts of tomatoes that RNA of interference, both
the derivatives from the transgene, and the Cuban
TYLCV are possible targets.
Development of bioinformatic tools
The work continued on the development of propietary
bioinformatic tools, such as: BisoGenet and Bisofarma, to
generate and analyze biological networks.
• Analysis of data from cell growth inhibition experiments
BisoGenet
• Classification of cell lines according to their sensitivity
• Implementation in the web site of a new algorithm of
in 60 cancer cell lines.
and the diseases selected for evaluation.
CIGB-552
• Visualization in surface response charts, of the synergic,
additive or antagonistic effect of medicine combination
experiments.
expansion of networks.
• Implementation of necessary functionalities to use the
new algorithm of expansion for the client.
• Readjustments in classes of the panel of visualization of
information, common to the BisoGenet and BisoPharma
applications, to support conditions not foreseen in the
original design.
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BisoPharma
• Incorporation of the R procedural language, which
allows to develop in the database functions that use this
powerful statistical package
• Development of stored procedures to determine the
enrichment of Genes in the category of chemicals
associated with illnesses.
• Development of stored procedures that allow to gain
access to the information about the types of targets, the
development phase of the drugs and the ways of action
of the drugs on their targets.
A preliminar version of the plugin of Cytoscape was finished
which generate and expand networks of genes, illnesses and
drugs, from a list of those mentioned entities given by the
user. In this version the user can select the types of relations
and primary sources of information wanted to incorporate
in the network, which can be included the entities given as
entry or those associated with them.
The nodes of the generated networks possess attributes
as type of node and description, which are visualized in the
panel of information of Cytoscape.
Proteomics
• A software was developed for the precursors' evaluation
in spectrums of masses of peptides interlaced by
cysteine bridges.
• Creation of a model for free quantification of non-labeled
proteins.
• Prediction of the limits of quantification of marking with
18O en experiments of proteomics, from the theoretical
model of oxygen exchange of the carboxylic terminal,
catalyzed by an enzyme.
In addition, the information was acquired and a methodology
achieved by the analysis of correlation of gene expression,
with data of inhibition of the growth of experimental cancer
cell lines.
Computer support
• Improvement in the infrastructure of the network giving
•
•
the biggest hardiness and safety to the computer
network.
Increase of the safety level and improvement of the
safety mechanisms of the network.
Modernization of the Telephone Slate, which has
allowed a significant progress both in the service and in
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the control of the calls. Introduction of the IP Telephony.
• Implementation of new services and maintenance of the
•
existing ones in the network of the CIGB that improve
the management of the work and the computer safety.
Support to the organization and development of the
different activities developed in the CIGB, such as:
Havana Biotechnology Congress, Audits, Visits, etc.
AWARDS AND distinctions
• Two researchers obtained the Doctor´s Degree in
Mathematics. Proposal of an annual of this tribunal.
• Two workers finished their carrier in informatics
engineering.
EVENTS
• International Workshop in Bioinformatics and OMICS: a
lecture and three posters.
• Technical and Scientific Forum at the center: Two
•
workers were presented:
- BisoGenet: a tool for System Biology. A Martin,
ME Ochagavia, J Miranda, J Fernandez-de-Cossio,
L Carmen Rabasa, C Suarez, F Torres, A Garcia,
R Bringas. Conferencia.
- BisoPharma: An integrated system for gene-diseasedrug network, reconstruction. ME Ochagavía, A García,
A Martin, F Torres, J Miranda, Y Grinión, J Fernándezde-Cossío, C Suárez, JR Fernández, R Bringas. Cartel.
- SysBiomics: Database for integration and analysis of
omics data. J Miranda, ME Ochagavía, A Martín, LC
Rabasa, C Suarez, J Fernández- de-Cossio, A García,
F Torres and R Bringas.
- Modeling the signal of enzimaticaly 18O labeled
peptides in the mass spectrum. J Fernandez-de-Cossio,
Y Ramos, Y Satomi, C Ferrero, M Fukuda, LJ Gonzalez,
T Takao.
Hematology 2013. Co-author of a poster.
PUBLICATIONS
An article was published.
SUMMARY
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Direction of Quality Control
T
he Direction of Quality Control at the Center for Genetic Engineering and Biotechnology
(CIGB) has as a principal mission to achieve analytical assays assessing the quality of
the developed products that are generated in the company; also to express analytical, exact
and reliable results. Besides, it verifies and implements all sampling and control procedures,
preparation, evaluation, maintenance, storing and custody of the Center’s reference
materials (RM) and assures the stability control of active pharmaceutical ingredients (API)
and final products (FP). It is also responsible not only to release or reject raw materials and
intermediate products but also to store and preserve working and primary cell banks.
As a main resource, this direction counts with a working staff devoted to a sum of objectives
and tasks to accomplish an efficient control system at the CIGB. This Direction promotes
the staff’s commitment and identification with the Center.
This Direction is committed with all its clients to guarantee efficiency and effectiveness of its
results, as a way of continuous improvement of all its processes.
The direction of Quality Control maintains a constant update of quality standards and
regulations applicable to the biopharmaceutical industry as a fundamental need for its
work’s projection, the CIGB quality products’ certification and the clients’ satisfaction and
trust.
To achieve these objectives, the Direction is composed by two departments and four groups.
DIRECTORA
MSc. Lourdes Beatriz Costa
Anguiano
[email protected]
Telephone: 271 6022, ext. 3210
Department of Biological products
The Department of Biological products achieves analytical techniques related with the
determination of in vivo and in vitro biological activity of all the CIGB’s products, quantification
and identification by means of immunological methods( ELISA, Western blot, Immunodot)
of APIs and host contaminant proteins; detection of host DNA contaminant; or methods of
molecular biology: hybridation, polymerase chain reactions (PCR), and storing and control
of primary cell banks are performed at this department. Moreover, it has a microbiology
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laboratory which is in charge of the microbiological control
and monitoring of processes, areas, and auxiliary systems
of the production plants.
Physical-Chemistry Department
The Physical-Chemistry Department develops the
purity’s identification and control methods of active
pharmaceutical ingredients (API) through high effectiveness
of liquid chromatography, gaseous chromatography and
electrophoresis, isoelectric focusing and peptide mapping.
Besides, proteins, additives and preservatives are quantified
in active pharmaceutical ingredients (API) and final products
by means of spectrophotometric methods. Raw materials
are controlled and released as well as physical and chemical
properties of the auxiliary systems for the productions. This
department is the responsible for the management of the
CIGB’s analytical validation techniques, the Quality control
audits of Good Laboratory Practices(GLP) in the Direction of
Quality Control and other CIGB’s laboratories.
Group of Stability and Reference Materials
The Group of Stability and Reference Materials of the
Direction of Quality Control is in charge of the stability studies
of all FP and APIs produced at the CIGB and registered by the
National Center for the Quality Control of Drugs (CECMED).
This group elaborates reference materials to be used in the
analytical techniques applied in all laboratories. This implies
characterization, homogeneity, and stability studies allowing
the establishment of each one
Besides, this group achieves tasks related to biostatistics
and automation of the necessary calculus in quality control.
Management and Costs Group
The Management and Costs Group is in charge of the
economic control: the productive and non-productive
services collection, the control of tangible fixed assets (TFA)
and tangible utensils and tools (TUT). It is also in charge of
planning and achieving purchases by consumption regulations
of the different analytical laboratories taking into account the
established quality specifications and investments.
Analytical Assurance Group
This group controls the compliance with good laboratory
practices, establishes and maintains procedures for the
control of documents which are part of the management
system of laboratories, establishes the central auditing system
and follows up corrective actions; ensures the competition
through training programs for the personnel operating the
equipments, makes assays and calibrations; and assesses
the results. Moreover, it guarantees to achieve validation of
analytical methods suitable for the foreseen goal.
Main Executives
Head of the Biological Product Department
MSc. Gladys Mabel Izquierdo López
[email protected]
Phone: (53-7) 250 4383
Head of Physical-Chemistry Department
MSc. Galina Maria Moya Fajardo
[email protected]
Phone: (53-7) 250 4395
Group of Analytical Release
Main Results
The Group of Analytical Release receives and distributes
samples given by the Direction of Quality Control to be
analyzed in different labs at the CIGB. In turn, it receives
the analytic results from the laboratories and makes up the
analytical dossier of each product lot. Besides, the group
is responsible for the reception, preservation, control and
destruction of the final products’ witness samples.
All these operations are complied with standard operation
procedures, validated methods, qualified equipment, and
traceable reference materials to guarantee the fulfilment of
the Good Laboratory Practices and international regulations
ruled by the control laboratories’ work.
• The effective analytical response has guaranteed
•
•
•
deliveries of products requested for the national network
and the main exports A decrease of 38 % in average
time of analytical release
Satisfactory results of the sterility area with positive
impact on the essay’s quality and time.
Technical effective response of Quality Control allowing
the obtainment of the certificate of Good manufacturing
Practices to the Praxis facility.
Demonstration of the traceability and stability of all the
RMs used in the development, process control and
quality control of Heberprot-P®.
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• Efficient strategic design to give analytical response
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in vivo and in vitro for the products Heberpenta®,
Heberpenta®-L and Gavac® without repetition of essays.
Approval of 3 kits with using commercial reagents, as
an analytical alternative for the quantification of protein
contaminants of the host.
Approval of the in vitro potency essay as an alternative
to the potency essay for the hepatitis b in the combined
vaccine Heberpenta®-L and Heberpenta®.
Approval of the validity criterion change of the
immunogenicity test of Haemophilus influenzae type b
(Hib) as part of the analytical method improvement.
Suitable analytical insurance of biological reagents, RM
and inputs.
Intense work with satisfactory results and delivery
on time of the information for actions of record and
commercial activities.
Satisfactory results in national and international audits.
Consolidation of the analytical battery for the control and
characterization of Heberprot-P® based on requirements
of EMA.
Increase of the quality standard of 14 RM with impact in
registered and new products.
Solution to the replacement of the lot of RM of
recombinant streptokinase in the method of chromogenic
Substratum, which allows to evaluate the lots for this
method in the Biocen facility and the product in the
Direction of Technological Development of the CIGB.
Technical assistance during TT essays of CC production
PRP in ChangHeber, China and technical exchange on
Hib for three records in Russia for Microgene.
Guarantee of safe water in the CIGB sanitary Control of
the drinkable water in 35 points that give the CIGB, CIB,
Heber Biotec S.A., buildings.
Demonstration of the stability of Heberprot-P®: API and
PT, after freezing and cycles of temperature up to the
implemented due date
Demonstration of the stability of Heberpenta®-L vaccine
for 36 months.
Demonstration of the storage temperature change of
API of the recombinant P64k, of -70 ± 50 ºC to -20 ±
50 ºC in hydrolytic glass flasks, with lid of polypropylene
thread and up to 12 months.
The stability study of API of IFN-ϒ up to 6 months was
completed.
Checking with industrial lots of the approved stability of
the Quimi-Hib® vaccine adyuvanted and the suppository
of the recombinant streptokinase.
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• Extension of the due dates of:
- API of Heberprot-P® from 6 to 12 meses.
- FPs of IFN-ϒ up to 18 months.
- Matriz of plantibody, of 24 months at 5 ± 3 ºC.
Awards and distinctions
XVI Forum of Science and Technology 2013
• Qualification of the transfer of samples that must be
transported of 2-8 °C. Mention.
• Essential elements of the organoleptic properties of the
•
•
Heberprot-P®. Prominent.
Starting up of the area of Sterility. Prominent.
Technological transference of the essay of Biological
Activity to Plant Vitoria, Spain, for the liberation of the
lots of Heberprot-P®. Prominent.
COURSES MANAGED AND DELIVERED
BY THE AREA
The personnel of Quality Control have participated as
teachers in different subjects of Master’s degrees and
courses:
• Courses and conferences given in the modules of
Quality Control for the Master's Degree of Trends in
Modern Biotechnology, CIGB
- Course of Methods of the Statistics Applied in
Biotechnology.
- Course of Stability of Pharmaceutical Products.
• Delivered course in the Master's degree of Engineering
of the Biotechnological Processes, ISPJAE.
- Stability of medicines.
Foreign visitors
Cláudia Maria da Conceição y Ozeias de Lima Leitao, Health
Ministry of Federal Government FIOCRUZ, Brazil. Training
in Reference Materials and Stability. Training in the use of
the Reference Material in the Orcinol method.
SUMMARY
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Direction of Quality Assurance
T
he mission of the Direction of Quality Assurance at the CIGB is to design and
implement the quality management system. Its main objective is to guarantee
quality, safety and efficacy in researches, services and developed products through the
compliance of the current international guidelines of Good Laboratory Practices.
This direction is committed to their clients (internal and external) to favor efficiency and
efficacy of the results; for the continuous improvement of the processes.
The Quality Assurance Direction has two Departments: Inspection and Audits and Quality
Improvement and Engineering; also four Groups, as follows: Metrology, Lots Release,
Documentation, and Administration and Costs.
DIRECTOR
Ing Yair Quiñónez Maya
[email protected]
Phone: (537) 250 4375
Department of Quality Improvement and Engineering
• It designs and implements the quality management system.
• It guarantees the management of processes and programs: the environmental
•
monitoring of clean areas and assessing the analyzes of the results, validation /
qualification, change control, complaints, and after-sales services to enquires from
external clients, withdrawals of products from the market, devolutions, purchasing
system of equipment.
It coordinates actions for reviewing the annual product and the system by the
direction.
Deparment of Inspection and Audit
• It provides the Quality Auditing system, which includes audits to the Quality
management system, clinical and preclinical studies, processes, suppliers of raw
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•
•
•
material and its components, contracted entities of
manufacturing and testing services.
It arranges the management of non-conformances and
the program of corrective and preventive actions.
It arranges the risk management .
It arranges inspections of quality: Inspect the productive
areas to verify the compliance with good manufacturing
practices (GMP), quality control, and inspection to the
documentation of deliveries and shipments, as well as
authorizations for campaign changes in multiproduct
plants.
Group of Documentation
It establishes the guidelines that allow the functioning
of the documentation system in the center, a key pillar to
ensure compliance with good practices. The documentation
system comprises the Central Group for Documentation in
the Direction of Quality Assurance ; as well as groups and
documentation activities in each of the plants and areas.
Lot Release
It ensures batch releasing of active pharmaceutical
ingredients and final products based on an integral review
of lot files which allow to guarantee product output to the
national distribution network and abroad with the required
quality. The group also arranges the documentation
accompanying shipments according to the client’s request.
Moreover, the group releases lots out of Cuba, through the
contracts of services and the compliance of the requirements
for the center, and the statement of the sponsors in the
specific contracts.
Metrology
It guarantees calibration and control of the measuring
instruments according to the Annual Calibration plan of
the different plants and areas. All work (and contracting
services) carried out by this Group comply with the good
manufacturing and laboratory practices. The group is in
charge of guarantying the traceability of the measurements
through national and international standards.
purchases of materials, control of tangible fixed assets(TFA)
and tangible tools and utensils (TTU).
Main Executives
Head of the Inspection and Audits Department
MSc. Inés Heredia Revilla
[email protected]
Phone: (53-7) 250 4390
Head of the Quality Improvement and Engineering
Department
MSc. Kenia María Vázquez Montero
[email protected]
Phone: (53-7) 250 4373
Head of the Group of Metrology
Tec. Leonel Godinez Cumbrado
[email protected]
Phone: (53-7) 250 4360
Head of the Documentation Group
Eng. Ana M. Cinza González
[email protected]
Phone: (53-7) 250 4376
Head of the Lot Release Group
Tec. Carmen Diana Pérez Pérez
[email protected]
Phone: (53-7) 250 4390
Head of the Group of Administration and Cost
Tec. Luis Álvarez Álvarez
[email protected]
Phone: (53-7) 250 4391
Main Results
• Certification of the productive system Plant 4 by Cecmed.
• Certification of the temporal production of the active
•
Administration and Costs
The group is in charge of coordinating the activities related
to fees and expenses of the direction, investments and
•
pharmaceutical ingredient(API) of Haemophilus influenzae
(Hib in the Biocén center by Cecmed).
Certification of the productive system of Plant 1 (Hepatitis
B); Plant 6 and 7 (IFN-PEG + epidermal growth factor
(EGF), parentheral; Plant 9 (factor de transference); Plant
10; ChangHeber, China (IFN, IFN-PEG) by Cecmed.
Keeping up the Certification the Quality System by ISO
9001:2008.
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• Satisfactory results in the Audit Trillium for the clinical studies of
•
Heberprot-P®.
Maintenance of the lot release system permitting to achieve all
production plans and planned sales.
International Projects
• Technical Committee Brazil–Cuba (human recombinant interferon
alpha 2b : hu-rec IFN alfa 2b).
• Advising the project to build a new vaccine manufacturing facility
•
•
•
highly specialized on infrastructure, equipment, and human resources
from Venezuela. National Institute of Hygiene “Rafael Rangel”.
Strategy to make export sales actions of liquid Heberon from JVC
ChangHeber, China, to Pakistan.
Strategy for the production and marketing of interferon alpha and
peg-interferon in the Popular Republic of China ChangHeber.
Project with Sinergium, pentavalent and hexavalent vaccine.
Events and courses
• A course based on Good Manufacturing Practices.
• Seminar about Quality System, after a received course in India.
• Seminars of Standard operational procedures of Documentation and
Metrology groups.
• To coordinate and deliver the conferences on compulsory subjects
•
•
•
of the quality mention of the Master's Degree of Trends in Modern
Biotechnology:
- Basic aspects of quality and tools. Quality System.
- Quality Assurance on biotechnological productions.
Participation in the Technical and scientific Forum at the center, as a
member of the tribunal.
Conference in the course Validation, in the Master's degree
biotechnological Processes.
The requalifications of all the procedures of the documentation
system were achieved that were changed, as well as in the system
of metrological assurance and lot release.
SUMMARY
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Human Resources Direction
T
he Human Resources at the Centre for Genetic Engineering and Biotechnology (CIGB)
is aimed to promote the continuous improvement of the organization based on strategies
for human resource development, so as to be able to develop research and production.
Continuous training and development of human capital are key pillars to meet the demand
of knowledge required, with superior quality, and ensure compliance with safety standards
and occupational health.
This Department has a strategic character and it’s included in the overall plans of the
organization. Among its main tasks is to ensure and fulfill planning, incorporation, selection,
placement, training, evaluation and compensation of human resources. This area is
responsible for the implementation of the existing legislation on wages, employment, social
security, health and training.
DIRECTOR
Oswald Ravelo Mallon
[email protected]
Phone: 250 4562; 271 6022 / Ext.
1189
Its specific roles and responsibilities are:
• Organize, direct and control the activity of training of workers and the board, from the
•
•
•
•
learning needs; select candidates and coordinate training programs; organize and
control the activity of social security and health at work; so as measures to preserve
the environment;
In conjunction with the labor union, it organizes the workers stimulation system in
accordance with their results in the services.
Propose ideas and participate in the programs of recruitment and selection of staff,
through established procedures.
Recruit staff and control probationary workers, retirees and contract workers.
Reconciling staff fluctuation, employment registration and cancellations, adjustments
and changes in the occupational categories in the areas and the Central Service Office
for Scientific Centers of the Ministry of Science Technology and Environment (Citma).
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YEARBOOK 2013
• Controlling disciplinary sanctions imposed on workers and ensure
their rehabilitation.
• Arrange licenses, certificates, medical reports, accidents and
pensions to workers.
• Comply with current legislation to ensure successful performance
•
•
•
•
•
evaluation, classification and valuation of jobs, and analyze wage
stimulation and salary administration.
Promote a favorable working atmosphere facilitating the development
of the staff, from a social, cultural and working point of view, and their
identification with the institution.
Guarantee everything about the assurance of workers' wages
from proper preparation and making of the payroll for all workers;
completing the procedures established by the Ministry of Finance
and Prices.
Promote and maintain the highest degree of physical, mental and
social well-being among workers of all professions, prevent damage
to health caused by working conditions, protect employees from risks
arising from the presence of harmful agents as well as place and keep
the worker in a job which is in accordance with their physiological and
psychological skills. In short, it seeks to adapt work to man and each
man to his job.
Working in conjunction with technological and scientific committees
of categorization for the promotion of workers
Ensure compliance with labor policy.
Key executives
Head of the Training Department
Ada Triguero Cruz
[email protected]
Telephone: 250 4172
Head of the human resources department
Mirlen Díaz Cárdenas
[email protected]
Telehone: 250 4111
Main Results
• ISO 9001:2008 certification was attained.
• Two workers of the area are included in the Heberprot-P® promotion.
• Satisfactory Development of the payment systems (Resolution
9/2008) and payment in convertible currency that are applied on
permanent basis. The payroll payment in local currency has been
attained between the 5th and 7th of each month, allowing advances
in the financial closures and delivery of statistical information in time.
Running of the process of scientific and technological categorization
including redevelopment of the latter
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YEARBOOK 2013
• Satisfactory participation in inspections and external audits.
• Program activities for the working people.
• Implementation of medical checkups of occupationally exposed and
•
•
•
•
unexposed workers and specialized tests to workers who work with
organic solvents, the fumigator and the ones who work with lead.
Furthermore nasopharyngeal swabs were made to workers directly
linked to production and Vivarium.
Professional drivers are reclassified according to the new Law on
Roads and Traffic.
There was a systematic attention to children during their summer
period.
Successful implementation of training activities and development.
The Status of Renowned Center at Municipal and Provincial levels in
the Forum of Science and Technique is maintained for an eight-year
period.
SUMMARY
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YEARBOOK 2013
HEBER BIOTEC S.A.
H
eber Biotec S. A. is the commercial company that owns the exclusive rights for
commercialization of the R+D projects, biotech and pharmaceutical “high-tech”
products; technological services; and the intellectual property rights which contribute to
social welfare of the Cuban population and to preserve the environment.
The commercial strategy involves maintaining the results in the current established markets,
working on the introduction into the most regulated markets of developed countries or the
first world and multiplying the company’s contribution to the Cuban economy.
Heber Biotec, S.A. portfolio by
•
•
•
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•
MANAGER
MSc. Madaisy Cueto Sánchez
[email protected]
Phone: (537) 250 4528
Heberfarma: Medications and vaccines for human use.
Heberdiag: reagents and diagnosis kits
Hebervet: Products for veterinary use.
Heberplant: Agricultural products.
Hebertec: Technological transfer.
Suppliers
•
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•
•
•
•
Center for Genetic Engineering and Biotechnology (CIGB)
AICA Laboratories
Novatec Laboratories
Laboratorios Liorad
Placental Histotherapy
Biocen
Cidem
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YEARBOOK 2013
HEBER BIOTEC S.A.
Main Executives
• Heberon Alfa R®: Recombinant Human Interferon Alpha
MSc. Dora García Delgado
General Vice-Manager
[email protected]
Phone: (537) 250 4528
• Heberon Alfa R® líquido: Human recombinant Interferon
Main Results
• The enterprise exported products to more than 35
•
•
•
Alpha 2b. Injectable liquid solution.
• Heberkinasa®: Recombinant streptokinase without
albumin against the myocardial infarction.
• Hebervital®: Granulocyte-Colony stimulating (G-SCF).
• PEG-Heberon®: recombinant human Interferon alpha
2b, conjugated to polyethylene glycol.
countries, with a 25 % sales increment compared to 2012.
Placental-derived products
commercialized by the enterprise. The products and
vaccines of the Center for Genetic Engineering and
Biotechnology (CIGB) positively impacted in the
treatment of 28 diseases which affects the Cuban
population.
The strategic alliances were strengthened with
commercial partners in China, Brazil, Russia, Vietnam,
Argentina, Ecuador, Venezuela and South Africa, and
other countries.
The Enterprise kept the ISO 9001:2008 certification for
its quality management system.
Five new sanitary registrations were granted to
enterprise products overseas.
A high visibility for the CIGB, Heber Biotec S. A. and their
products was attained through a wide press coverage,
events attendance and the availability of different
promotional formats.
• Melagenina Plus®: Treatment for vitiligo.
• The national demand was covered for the medicines
•
2b, antiviral, antiproliferative and inmunomodulatory
Product Portfolio
Heberfarma
Vaccines
Generic Drugs
Antihypertensive
• Amlodipine 10 mg, tab. (Anticalcic)
• Verapamil hydrochloride 5 mg, inj. (Anticalcic)
• Captopril 50 mg, tablets (inhibitor of the angiotensin
convertase enzyme, ACE)
• Enalapril 10 and 20 mg, tablets (ACE inhibitor)
• Carvedilol 6.25 and 12.5 mg, tab. (α and β blocker)
Analgesic and hypnoanalgesic
• Tramadol 100 mg, inj.
Analgesic and antipyretic
• Acetyl Salicylic Acid 500 mg, tab.
Anaesthetics and anesthesic adjuvants
•
• Heberpenta®-L: Pentavalent vaccine against the •
diphtheria, tetanus, whooping cough, hepatitis B and •
haemophilus influenzae type B.
•
• Heberbiovac HB: Recombinant vaccine against •
antihepatitis B.
•
• Quimi-Hib®: Conjugated vaccine against Haemophilus •
influenzae type b.
•
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Biological Pharmaceutical products
•
• Heberprot-P®: Product that favours healing of diabetic •
•
foot ulcers and reduces the risk of amputation.
Bupivacaine 0.5 %, inj.
Ketamine 10 and 50 mg, inj.
Lidocaine hydrochloride 2 %, 20 mL, inj.
Lidocaine 2 %, inj.
Procaine 2 %, inj.
Atracuriu besylate 25 mg, inj. (anesthesic adyuvant)
Pancuronium Bromide 4 mg, injection (anesthesic adjuvant)
Fentanyle 0.05 mg, injectable (anesthesic adjuvant)
Mephenesin 500 mg, tablets (muscle relaxant)
Midazolam 10 and 15 mg, injection (muscle relaxant)
Pancuronium 4 mg, SP (muscle relaxant)
Succinylcholine 50 mg, i injection (muscle relaxant)
Lido 2 %-Epin1:50000, carpule (dental anesthetic)
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YEARBOOK 2013
HEBER BIOTEC S.A.
• Lido 2 %-Epin1:80000, carpule (dental anesthetic)
• Mepivacaine 3 %, inj. (dental anesthetic)
Inotropic Agents
Antihypertensive and Antianginal
• Dobutamine 250 mg, injection
• Dopamine 200 mg, injection
• Diltiazem 25 and 50 mg, inj.
Anxiolytic
Benzodiazepine antagonist
• Diazepam 10 mg, injection
• Flumazenil 1 mg, injection
Anti-hypoglycemic
Platelet antiaggregate
• Dextrose 20 and 50 %, injection
• Clopidrogel 75 mg, tablets
Antihistaminic
Non-steroidal anti-inflammatory
• Diphenhydramine 20 mg, injection
• Loratadine 10 mg, tablets
• Diclofenac 75 mg, inj. (analgesic)
Anti-ulcer
• Cimetidine 300 mg, injection
• Omeprazol 20 mg, tablets, and 40 mg, injection
• Ranitidine 50 mg, injection
Antifungal
• Fluconazole 150 mg, capsules
• Miconazole 200 mg, injection
Anti-migraine
Antibiotics
• Sumatriptan, injection
• Azithromycin 250 capsules and 500 mg, tablets
• Gentamicin 10 and 80 mg, injection
• Kanamycin 50 and 100 mg, inj.
Anti-osteoporotic
Anti-infective Agent
Anti-psychotic
• Sodium Sulfadiazine 10 %, injection
• Risperdone or Risperdal 3 mg, tablets
Anticoagulant
Anti-retroviral
• Heparin 25 000 U, injection
•
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Anti-diabetic
• Glimepiride 1 and 4 mg, tablets
Antiemetic
• Dimenhydrinate 10 and 50 mg, injection
• Ondansetron 4 and 8 mg, injection
• Zolendronic Acid 4 mg, injection
Stavudine 40 mg, cap.
Indinavir 200 mg, capsules
Lamivudine 150 mg, tablets
Nevirapine 200 mg, tablets
Triviral 30 mg, tablets
Zidovudine 100 mg, capsules
Anti-asthmatic
• Montelukast 5 and 10 mg, tablets
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YEARBOOK 2013
HEBER BIOTEC S.A.
Bronchodilator
Laxative
• Aminophylline 250 mg, injection
• Fenoterol 0.5 mg, injection
• Bisacodyl 5 mg, tablets
Antiviral
• Ribavirine 200 mg and 400 mg, capsules
Cardiotonic
• Digoxin 0.5 mg, injection
Neuroleptic
• Haloperidol 5 mg, injection
Moisturizing Solution
• Dextrose 5 %, injection
Vasoconstrictor
Corticosteroid, anti-inflammatory and anti-allergic
• Norepinephrine bitartrate 8 mg, injection
• Betamethasone 4 mg, injection
Vasoconstrictor, bronchodilator
Diuretic and osmotic
• Epinephrine 1 mg, injection
• Mannitol 250 mg, injection
Peripheral vasodilators
Electrolytes
• Pentoxifylline 300 mg, injection and 400 mg, controlled
released tablet.
• Calcium chloride10 %, injection
• Potassium chloride 1.8638 g injection (equivalent to 25 Vitamins
mmol (mEq) K+)
• Calcium Gluconate 10 %, injection
• Calcium Gluconate 0.30 g, injection (equivalent to 1.28
mmol (mEq) K+)
• Magnesium Sulphate 10 %, injection
Benign prostate hypertrophy
• Terazosin 2 and 5 mg, tablets
• Hidroxocobalamine 0.1 and 1.0 mg (vit. B12), injection
• Pyridoxine hydrochloride 25 and 50 mg (vit. B6), injection
• Thiamine hydrochloride 100 mg (vit. B1), injection
HEBERDIAG
Diagnostic Kits
• AuBioDot anti-gliadin: For the detection of antibodies in
human serum against gliadins.
Hypnoanalgesic
• HeberFast Line pregnancy: For early diagnosis of
• Morphine Hydrochloride 10 and 20 mg, injection
• Pethidine 50 and 100 mg, injection
• HeberFast Line MaterniTest: Self-experiment for the
Lipid lowering drug (Statin)
pregnancy in urine.
early diagnosis of pregnancy in urine.
• HeberFastLine Rotavirus: A step and quick test, for the
detection of Rotavirus in feces.
• Atorvastatin 10 and 20 mg, tablets
Hebervet
Oxytocic
Veterinary
• Oxytocin 10 U, injection
• Gavac®: recombinant vaccine against ticks. Boophilus sp.
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YEARBOOK 2013
HEBER BIOTEC S.A.
Aquaculture
• Acuabio 1: nutritional supplement for aquatic organisms.
HEBERPLANT
• HeberNem®-S: biological product for the control of plant parasites
nematocides.
HEBERTEC
• Transfer of Technologies developed by the CIGB for the production
of recombinant drugs
• Joint Research-Development projects.
• Development and scaling of the production processes of recombinant
products.
• Diverse technical services, consulting and staff training in modern
biotechnology field.
Participation
workshops
in
fairs,
expositions
and
• IX Science Workshop y X Pedagogical Workshop of the Latin
American School of Medicine (LASM).
• V Cuban Earth Sciences Convention and Exhibition, Geociencia
2013.
• XXIII Congress of Internal Medicine of Central America and
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•
•
Caribbean, Internal Medicine 2013.
VII Cuban Congress of Hematology 2013.
ExpoSalud Ecuador.
I Festival of Social Communication.
XIV International Congress of the Cuban Society of Orthopedics and
Traumatology, Orthopedic 2013.
LatinFarma Havana 2013.
XX Cuban Congress of Urology 2013.
XXXI Internacional Fair, FIHAV 2013.
I Cuban Symposium of Cardiovascular Death 2013.
XXIII Meeting of the ALPA and IV Congress of Animal Production.
ANAP, ExpoCuba.
Visitors
Among the 335 visits received, 249 were foreigner and 2459 Cuban.
SUMMARY
81
YEARBOOK 2013
INTELLECTUAL PROPERTY DIRECTION
T
he Intellectual Property Direction of the Center for Genetic Engineering and
Biotechnology (CIGB) is committed to ensuring the protection of all new results of the
institution, through the modality of intellectual property (IP) best suited for each case and
guides the activity of research and development (ID) to high novelty and inventive steps,
which generates high-impact results on the market and thus achieve better use of each
product and project generated at the CIGB.
Intellectual Property Policy at the CIGB
Raimundo Ubieta Gómez
[email protected]
Telephone: (537) 250 4440
• IP strategy for each research project.
• Periodic analysis of the results of R & D for patent protection or other form of IP
•
•
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•
•
protection.
Publication strategy defined according to the characteristics of each project.
Using patent data as a source for updating the state of the art and strategic analysis.
Using the know-how as a protection of intangible resources.
Using technology transfer as product.
Do not sell intellectual property. Use other forms of patents negotiation.
The patents portfolio of the CIGB has 60 inventions and 1237 international patents and
international applications distributed in major markets worldwide. Over 80 % of them in
countries of the first world (USA, Europe, Japan, Canada, etc. and in the so-called emerging
countries. 68.5 % of the patents are granted (Figure 1).
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YEARBOOK 2013
INTELLECTUAL PROPERTY DIRECTION
Figure A
> 40 patents
21 - 39 patents
10 -20 patents
< 10 patents
Figure B
Rest of the World. 20
First World. 53
BRICS + AR + KR + MX. 27
CIGB distribution patents worldwide. A) Geographical distribution. B) Distribution by groups of countries. First World:
USA, European Patent Office, Canada, Japan, Australia. AR: Argentina. KR: South Korea. MX: Mexico. BRICS: Brazil,
Russia, India, China and South Africa.
SUMMARY
83
YEARBOOK 2013
Business Development Group
T
he Business Development Group (GNDP) is a Department of the Center for Genetic
Engineering and Biotechnology (CIGB), attached to the General Direction, and is in
charge of the management and international negotiation of the research and development
projects in the biotechnology area.
It coordinates business development structuring and carrying out high novelty projects
from early stages until they become competent products in the markets of high regulatory
requirement.
Key Executives
DIRECTOR
Ing. Ernesto López Mola
[email protected]
Phones: 271 2397; 250 4120
Vicedirector
Dr. Ricardo Silva Rodríguez
[email protected]
Telephones: 271 2397; 250 4121
Executive Negotiation and Project Management
Miriela Gil Mena
[email protected]
Telephones: 271 2397; 250 4121
Executive in Management and negotiation of Agricultural Projects at the CIGB
Marianela García Siverio
[email protected]
Telephone: 250 4113
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YEARBOOK 2013
Executive in Management and Negotiation of projects
Dr. Boris Ernesto Acevedo Castro
[email protected]
Telephones: 271 2397; 250 4120
Heberprot-P® Promoter
Amelia de la Caridad González
[email protected]
Telephone: 271 6022, Ext. 1130
Ariadna Hernández García
[email protected]
Telephones: 250 4113
Yunarquis Fernández Reynaldo
[email protected]
Telephone: 250 4608
Jorge Martín Machado
[email protected]
Telephones: 271 6022, Ext. 1020
Erick Hernández Leyva
[email protected]
Telephone: 250 4113
Project Promotion
Webmaster
Mailyn Palmero Molina
[email protected]
Telephone: 250 4608
Secretary
Ivona Gutiérrez Masuet
[email protected]
Telephone: 250 4608
Promotion of international projects
Executive in Management and Negotiation of Projects
Dr. Manuel Raíces Pérez-Castañeda
[email protected]
Telephones: 271 23 97; 250 4121
Project Promotion
Dunia Martínez Arcia
[email protected]
Telephone: 250 4608
Israel Miranda Arnet
[email protected]
Telephone: 250 4113
Reina Lourdes Morejón
[email protected]
Telephone: 2716022, Ext. 1185
Karelia Macías Cosme
[email protected]
Telephone: 250 4113
Relevant Results
1- Follow-up to the national and international Program of
Integral Attention for patients with diabetic foot ulcers using
Heberprot-P® therapy:
• The resulting figures for patients inclusion approved by
the Ministry of Health of Venezuela was overachieved
for five consecutive years: 25 thousand patients with
diabetic foot ulcers (37 485 at the end of week 49).
110 417 patients have been attended with Heberprot-P®
therapy (week 49).
• Export plan (invoices) approved for 2013 to Venezuela
was overfulfilled.
• The integral Care Program with Heberprot-P® therapy
for patients with DFU was consolidated in 17 states
of Venezuela. Efforts are been made to implement
it in three more states. The structure and the working
system of the project management was consolidated in
Venezuela: Mission Barrio Adentro Mission, angiologists,
promoters, national tours, weekly reports, among other
key elements in the success of the program.
• The plan including Cuban patients in the program with
therapy Heberprot-P® therapy was overfulfilled: 9368
(end of week 50), representing 134 % of the annual
compliance plan (7000 patients). The promotional
structure was consolidated.
• The protocol for the integral program with the use of
Heberprot-P® in diabetic patients was consolidated in
85
YEARBOOK 2013
•
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•
•
all services of Angiology in Cuba. Efforts were made
in developing the protocol in the polyclinics. Three
routes and three meetings with angiologists and Cuban
specialists for evaluating program outcomes were
performed.
A new business agreement with the Ecuadorian Institute
of Social Security, to the use of Heberprot-P® in Ecuador
was signed.
Two national meetings were held: the Third Meeting of
the National Group of Angiology and the one with the
Central Unit of Medical Cooperation (UCCM) and the
main leaders from the provincial health directorates
(DPS), the Ministry of Public Health (Minsap), considered
of a great impact in the national program.
Working relationships with the National Angiology
Group, National Institute of Angiology and Vascular
Surgery (INACV) and the departments of Health Care,
Human Resources, International Relations and with the
Postgraduate Department of Minsap were consolidated.
Nearly 150 conferences on diabetes were taught to
promoters and executives in Cuba, Ecuador, Venezuela,
Dominican Republic and other countries, as well as
foreign dignitaries who visited the CIGB.
More than 200 news were broadcasted on television,
the radio or the press. Meetings with diabetic patients
and their relatives were made.
The national workshop, Echoes of the Congress, was
developed in every Cuban province, where results and
impact of the Second International Conference on the
integral program for patients with UPD with the use of
Heberprot-P® 2012 were stated, with the participation of
provincial health executives and specialists.
We participated in Mesa Redonda television progarm in
April.
We participated in the Festival of Social Communication
with a lecture on the Heberprot-P®.
Workshops and national and international courses on
Heberprot-P® were prepared and done.
The twelfth and thirteenth national meetings on extension
of Heberprot-P® use were successfully implemented on
May and December 2013.
The third and fourth international workshops on the use
of Heberprot-P® in patients with UPD were successfully
conducted.
The 2nd edition of the Advanced Course on Heberprot-P®
was performed.
The number of visits to Heberprot P® website was
exceeded in more than 1 million.
• The brochure for the international event “Managing
diabetes and its severe complications." was made. The
event was promoted. A press conference was offered.
• There was a follow up to the Heberprot-P® promotion
campaign.
2- Outstanding job of negotiating and signing of several
business agreements and projects of CIGB.
• Significant progress was made in promoting and
negotiating new projects at the CIGB. Projects were
signed.
• NASVAC® with Wittycell, France: For the European
Union, Japan, Southeast Asian countries and other
territories.
• NASVAC® with Generium, Russia: Russia, Belarus and
Kazakhstan.
• HPV with Generium, Russia: Russia, Belarus and
Kazakhstan.
• NASVAC® for Bangladesh.
• Quimi-Hib Plant in China was opened, negotiation and
starting of supply of PRP for the CIGB to ensure the
production plan of the pentavalent vaccine Heberpenta®
for 2014.
• Incomes were achieved by the signing of PEG-IFN
agreement (China).
• Incomes were achieved by the project GCSF-PEG 9
(China).
• Incomes were achieved from Negotiation agreement
projects signed or in progress.
• Significant progress in the negotiation of agricultural
projects at the CIGB.
- Creation Process of EM-Heber Lukang.
- The company Koppert Biologicals was identified as the
counterpart to other territories. Contract in discussion.
- The application for registration in Morocco, as intrant
Agricole and in Saudi Arabia, was presented as a
biopesticide.
- Evaluation agreement of Gavac® was signed in Korea.
- Evaluation agreement of Gavac® was signed in Costa
Rica.
Institutional Achievements
• Surpassing of the national inclusion plan in the
•
Integral care program of DFU patients with the use of
Heberprot-P®.
Surpassing of the national inclusion plan in the
Integral care program of DFU patients with the use of
Heberprot-P® in the Bolivarian Republic of Venezuela.
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YEARBOOK 2013
• Fifth anniversary of the Integral Care Program of DFU patients with
•
•
•
•
•
•
the use of Heberprot-P® in the Bolivarian Republic of Venezuela: 5
years of great efforts and satisfactions.
Negotiation and signing of an agreement with the Ecuadorian Social
Security Institute (IESS) for the implementation of the Integral Care
Program of DFU patients with the use of Heberprot-P® in 2014.
Successful completion of the implementation of promotional work
and inclusion of DFU patients in the Integral Care Program with the
use of Heberprot-P®, Ministry of Public Health in Ecuador.
Negotiation and signing of an agreement with the IESS for the
implementation of the Integral Care Program of DFU patients with
the use of Heberprot-P® in 2014.
Outstanding work of national and international promotion of
Heberprot-P® during 2013.
Outstanding job of negotiating and signing of several product
business agreements and projects of the CIGB.
Significant progress in the negotiation of agricultural projects at the
CIGB.
COURSES AND EVENTS MANAGED AND DELIVERED
BY THE AREA
• Training of 40 specialists from 15 countries in the integral management
•
•
•
of diabetic foot with the use of Heberprot-P® to satisfy mission abroad.
Training of the human capital included in the Program (1 Training
Promoters, 2 editions Diploma No. 1, Issue of Intensive surgical
procedures, 1 Advanced Course on the use of Heberprot-P®, 3 Intensive
Course on Diabetes). It included the preparation of the 5 Working
Groups selected to Ecuador by the IESS, preparation of 8 specialists
and 7 promoters incorporated into the Program in Venezuela.
Arrangement of two international courses on practical procedures
in the management of patients with diabetic foot ulcer using
Heberprot-P®.
Teaching on Cuban Biotechnology Updates and one on the integral
management to patients with diabetic foot ulcers to more than 13 000
Cuban doctors and medical aid workers throughout the year. That
includes the 6000 Cuban medical aid workers who are in Brazil.
Foreign Visitors
Support in the organization and attention to more than 10 000 visitors
including VIPs as presidents, vice presidents, ministers and deputy
ministers who visited the CIGB in 2013.
Publications
Two articles were published in international journals.
SUMMARY
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YEARBOOK 2013
DIRECTION OF CLINICAL RESEARCH
T
he Direction of clinical research (DCR) of the Center for Genetic Engineering and
Biotechnology (CIGB) has as mission to achieve clinical trials (CT) and pharmacosurveillance (PS) of vaccines and drugs obtained in the Center. Therefore, it counts with a
group of professionals and technicians specialized and scientifically trained in all phases of
human research.
The primary activity involves three departments of CT with dedicated monitors to coordinate,
design and control the achievement and completion of protocols of clinical research (CR) in
Cuba and abroad. A group of VF is responsible for monitoring the safety profile of products
and during post-marketing use.
The Group of statistics guarantees the optimal management of the data of the test and
the corresponding statistical analysis; while the specialists contribute to the automation
of processes and activities such as advanced electronic designs for the remote entry of
clinical data.
A group of supplies is responsible for the distribution of pharmaceuticals and other
necessary inputs to the clinical sites. It is also responsible for the development, control and
management of the random lists, storage conditions and the traceable accounts of products
during the trials.
The Group of clinical laboratory works with biological samples of patients. It is in charge
of biochemical and immunological investigations very specific, not available usually in the
health system; mainly pharmacokinetic, pharmaco-dynamic and immunogenicity studies.
The management and Control section is in charge of the coordination of training workshops
and satisfy printing, accommodation, transportation services and other logistical actions for
monitors and patients during the CT.
The documentation and good clinical practices section achieves the compliance with
the quality policy based on the NC-ISO 9001:2001 and international guidelines of good
laboratory and clinical practices.
MANAGER
Dra. Verena Lucila Muzio González
[email protected]
Phones: (537) 208 7378,
271 6022 ext. 1250-1251,
208 0428 ext. 106
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YEARBOOK 2013
The services provided by the DCR are distinguished by their
effectiveness, efficiency, reliability, reproducibility, security
and quality, based on evidences for decision making and
mutually beneficial relations with suppliers and clients.
Managers and workers in the Direction are permanently
committed to increase satisfaction and comply with the
clients’ expectations.
The goal of the DCR consists of maintaining excellence and
leadership in the CR activity achieving the highest quality
standards, contributing to the development of the national
biotechnology industry with new therapeutic alternatives for
diseases that constitute health problems and insert the CIGB
products in the most demanding international markets.
Main executives
Head of the Heberprot-P® department
Dr. Julio Esmir Baldomero Hernández
[email protected]
Phones: (53-7) 208 7379, 208 0428, ext. 108
Head of the Department of Oncology and other projects
Hugo Nodarse Cuní
[email protected]
Phones: (53-7) 208 7379, 208 0428, ext. 107
Head of the Vaccine Department
Dr. Zurina Cinza Estévez
[email protected]
Phones: (53-7) 208 0428 ext. 133
• The Cuban regulatory authority approved an application
•
•
•
•
•
•
•
•
Main results
The DCR directly contributed to the annual strategic
objective of the CIGB aimed to develop new products and
markets according to the 2013-2017 Strategic Plan. The
clinical research activity participated in 195 results, 157
considered extraordinary in monthly evaluations by the Board
of Directors. The Scientific Council of the CIGB approved
12 scientific-technical achievements and 38 institutional
achievements in which some authors were members of the
DCI. The general results are:
• Sixty six objectives of clinical investigations, 43 included
in the plan related to the 13 portfolio pipeline of research
+ development (R+D) and 6 products registered. 16
medical indications were clinically investigated, in
the main lines of infectious diseases, cancer, healing,
cytoprotection and neonatal, among others.
•
•
to give permission for a new clinical trial and seven final
reports of CT were concluded.
All the proposed clinical trial (CT) to achieve in the
country received an approval for the Ministry of Public
Health.
Satisfactory results in international audits received
during the year.
Advances in CT have continued and definition of
strategies in the projects of R+D negotiated with first line
products, as CIGB-247, CIGB-300 and Heberprot-P®.
Requests were responded for clinical information for
obtaining or renewal of health records in Cuba and
abroad, with an emphasis on the main products of first
level occurred.
The stage of the treatment of patients at the national
extension of the use of the PEG-Heberon® for the
treatment of chronic hepatitis C concluded and the
inclusion of patients for treatment with HeberPAG® in
skin tumors not melanomas continued.
A third study was achieved that confirmed the impact
of the use of the Heberprot-P® in the reduction of the
amputation rate in health institutions selected in different
areas of the country.
Post-marketing information was consolidated, enabling
a better understanding of the safety and effectiveness
of the Heberprot-P® profile, and the collection of this
information at the national and international levels was
also advanced.
Satisfactory results were carried out in the audit by
the national Office for Standardization (NOS) and the
Spanish Association for standardization and certification
(Aenor), which granted re-certification of the quality
management system of the CT for the third consecutive
year from the certification achieved in 2009.
Scientific dissemination of clinical findings increased
in high-impact journals, with an emphasis on first-level
products.
The project portfolio of the CIGB was strengthened.
Heberprot-P®
• Satisfactory results in the external audit were carried
•
out by the Trillium company hired by Chemo for clinical
studies of Heberprot-P® in China and pharmacokinetics
75/225 µg PK-BD in Cuba.
Participation in the methodological process of selecting
and preparing of new promoters in the programme of
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YEARBOOK 2013
•
•
•
•
•
•
comprehensive care of patients with diabetic foot ulcer
treatment using Heberprot-P® in Cuba and Venezuela.
Participation of medical specialists in the advanced
course about diabetes prevention and treatment,
and comprehensive care of diabetic foot ulcer using
Heberprot-P®.
Demonstrative protocols using Heberprot-P® in the
treatment of diabetic foot ulcer were evaluated and sent
to eight countries. Technical and commercial missions
requested by the management of the project were also
achieved. The approval of the registration for the product
was obtained in Peru.
Conclusion of the third study: impact of the use of
Heberprot-P® at the amputations rate. Selected
institutions. Cuba, 2011. This study covered 2011 and
first semester of 2012 information, and also included
1373 patients with diabetic foot ulcer(DFU), graduates
of 8 health institutions located in 5 provinces of the
country. The results were useful for the contribution that
represents DFU with the total number of amputations
in the institutions and were taken as reference to
evaluate the impact of Heberprot-P® in the reduction of
amputations.
Reports of effectiveness and safety using Heberprot-P®
in three countries were achieved: Venezuela (397
patients), Uruguay (50) and Argentina (323).
The quantitative analysis of differential gene expression
49 of neuro-infectious and ischemic DFUs treated with
Heberprot-P®.
Validation of the ELISA system for the detection of the
epidermal growth factor (EGF) that detects with precision
and accuracy the EGF-51-52 contained in Heberprot-P®,
in the conditions of the clinical research laboratory of the
CIGB. Its development and validation was in accordance
with the EMA-FDA regulations and subsequently
applied successfully in the product pharmacokinetics,
clinical trial to demonstrate that the active substance is
stable in human plasma platelet-free after 12 months of
conservation. This ensures its use for the quantification
of EGF-51-52 in the different clinical trials that require
it and adhere to the conditions evaluated. The results
were distinguished in the international audits received
by the project.
CIGB-300
The completion of the intravenous application study
of the CIGB in solid tumors, for the first time allowed
the characterization of clinical outcomes of safety,
pharmacokinetics, and survival of the product use by
intravenous route. This information of great value for the
clinical product strategy, was presented, discussed and
recognized by all the participants of the workshop, where
the clinical feasibility was ratified to a safe scaling of doses
in the patient. The results were in accordance with what
was observed in the pharmacokinetics and the main events
associated with the mechanism of action of the CIGB-300 in
tumor cells.
CIGB-247
The first evidence of a sustained humoral and cellular
response was obtained specific for VEGF in human
beings, which allowed ratifying the continuity of the clinical
development of the product. A phase I second trial began in
advanced solid tumors (Centauro-2 study) that concluded
the inclusion phase for patients with seven months in
advance on the planned schedule.
Heberpenta®-L
In just three months the inclusion of 584 infants concluded
successfully in the seroprevalence study, together with the
sampling and implementation of survey, the evaluation of
more than 6 lots of the vaccine. This study provided valuable
information post-marketing on immunogenicity against the
five antigens of several lots of Heberpenta®-L in terms of
application according to the everyday practice by the national
programme of immunization in Cuba.
Pegylated Interferon alpha to PEG-48 kDa
The phase II-III study in Brazil began for the treatment
of chronic hepatitis C. A phase II CT was successfully
completed with 24 patients; 392 blood samples (total
planned) during 20 consecutive weeks, from Monday to
Saturday. A phase III CT was advanced to the inclusion
of 210 patients (28 % of the total) and demonstrated
the antiviral effect of the pegylate interferon to PEG-48
kDa on the hepatitis C virus (HCV) viral kinetics, which
clinically meant 76 % of early virologic response with the
product. This result was in accordance with the hypothesis
of non-inferiority against the planned PEGASYS. The
achievement of the trial received satisfactory audit reports
by the sponsor and ANVISA-Cecmed inspection at two
clinical sites.
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HeberPAG® - Heberferon®
Three different clinical trials began and more than 130
patients were included:
• BRATINC-II study of tolerance of the intravenous
application of the HeberFERON® in recurrent or
progressive diffuse brain tumors.
• SCIENCE study for evaluating the use of HeberFERON®
in parpebrales tumors.
• Incarbacel V study for evaluating various schemes of
treatment with HeberFERON®.
• Elaboration of three final reports and a fourth with the
opinion of the Cecmed approval.
• SOFIA study: pharmacodynamics of two formulations
(HeberFERON® and HeberPAG®) with synergistic mixture
of interferon in healthy volunteers. Approved by the
Cecmed.
• Report of patients with more than 3 months of treatment
with HeberPAG® in patients with advanced solid tumors
or metastatic without therapeutic option (SOLINCcomp).
Significant result of the increase in survival patients with
renal carcinomas at high-risk for over 30 months.
• Dose study of the perilesional CIGB-128 in basal skin
cell Epithelioma. InCarbacel III study: follow-up review of
the third year of cases who obtained complete response,
demonstrating the effectiveness of the product in long
term.
CECIN study: intralesional administration of HeberPAG®
in patients with squamous-cell skin carcinoma. Follow-up
report of the 1st year of the cases who obtained complete
response, which confirmed the effectiveness of the product
in the long term
• The patent for HeberFERON® was granted in USA and
Japan.
• Renewal of the registration of HebePAG® 3 M and 10 M
in Cuba.
• Renewal of the registration of Heberon Gamma R® 0.5
M IU in Cuba.
PEG-Heberon®
PEG-Heberon® and Ribavirin in the treatment of chronic
hepatitis C (CubaPEG-HC). The report of efficacy was
updated showing a significant reduction in baseline HCV
viral load in 43 % of the patients, after 4 weeks of treatment.
At week 12, it is reduced in 69 % and 48 % completed the
treatment with undetectable levels. The sustained virologic
response was 25 % by purpose to treat, which includes
56 % in the responders to treatment. These results have been
regarded as excellent and endorsed by gastroenterologists
linked with PEG - Heberon® prescription.
STATISTICS GROUP
In the year 2013, the statistics group was involved in 18
clinical projects, one agricultural and one biomedical.
The use of adaptive designs in several studies was the
most significant. In total there were 52 statistical reports,
distributed as follows:
• Final reports: 19
• Statistical analysis related to clinical trials: 12
• Design of clinical studies: 21
INFORMATICS GROUP
A new design of the website of the area was designed
and implemented, visible on the intranet of the CIGB. All
the information and available access were updated. The
implementation of the remote entry system of clinical data
was continued from a larger number of hospitals participating
in clinical research.
THE SUPPLIES’ GROUP
Availability, conservation, preparation and distribution of
products and supplies requested were guaranteed for
13 clinical investigations, in 41 clinical sites of 9 Cuban
provinces. 1419 requests were attended which meant a
significant increase of 358 requests (133.7 %) compared
to 2012, which constitutes an annual record for the Group's
supplies the compliance of good clinical practices in the
management of random lists and masking products was
ensured. Delivery immediacy of the resources requested
was achieved, no complaints or non-compliance on the part
of customers was recorded as successful.
During 2013, the masking of 8500 bulbs products intended
for clinical research abroad was assured. This is the highest
number achieved so far by this group since its creation.
The cold chain was validated during storage, transportation
and distribution of products in clinical research was made as
well as the analysis of risk at this stage.
A record of temperature control was designed and
implemented in clinical sites and nine professionals from the
Department of pharmacy were trained in compliance with
good clinical practice.
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GOOD CLINICAL PRACTICES AND
DOCUMENTATION SECTION
The scope of the self-inspection of good clinical practice was
extended to 11 clinical trials in six Cuban provinces.
•
SECTION OF MANAGEMENT AND CONTROL
There was a significant optimization in the processing and
logistics coordination for the trials and a notorious advance.
There were also improvements in the quality and quantity
of activities with a successful performance and high level of
clients’ satisfaction.
The processing and central control of the proper use of
means of transportation rented for the activity of clinical
trials, pharmaco-surveillance, and promoting Heberprot-P®
in Cuba were a success.
•
•
AWARDS AND DISTINTIONS
• Hugo Nodarse Cuní et al. Development of PEG-Heberon®
•
•
•
•
•
•
for the treatment of hepatitis C in Cuba. Provincial award
of innovation 2012 by Citma in Havana..
Hugo Nodarse Cuní and et al. Development of PEGHeberon® for the treatment of hepatitis C in Cuba.
Annual Health award by central and national authorities.
Isis Belkis Yero Alós et al. Heberprot-P® in diabetic foot
ulcer patients: active pharamaco-surveillance. Cuba,
may 2007-septiember 2010. Annual Health award by
central and national authorities.
Giselle Pentón Rol et al. Ficocianina project in multiple
sclerosis. Annual Health Award by the Central And
National Authorities.
Isabel Alicia Guillén Pérez and et al. (Angela Damary
Tuero Iglesias, DIC). Molecular mechanisms involved in
the inhibition of the proliferation of tumor cells exposed
to high concentrations of epidermal growth factor (EGF).
Annual award of the Academy of Sciences of Cuba 2012.
Julio César Aguilar Rubido et al. (Verena Lucila Muzio
González, DIC). Demonstration of safety and innovative
immunological properties related to treatment with the
therapeutic vaccine candidate against hepatitis B chronic
(Nasvac) in three models of transgenic mice expressing
the hepatitis B virus of or its surface antigen. Annual
award of the Academy of Sciences of Cuba 2012.
Francisco Hernández Bernal et al. Latin American
DAHER CUTAIT award: recombinant streptokinase
suppositories for the treatment of acute hemorrhoids.
•
XXIII Latin American Congress of Coloproctology. IV
International Congress of surgery Colorectal of ASCC.
San Salvador, El Salvador (July, 2013).
Francisco Hernández Bernal et al. Relevant result and
special distinction in the scientific and technical Forum,
Municipal and Provincial authoritative ranks: recombinant
streptokinase suppositories for the treatment of acute
hemorrhoids. Demonstration of efficacy and safety in
clinical trials. Cuba, 2006-2012.
Francisco Hernández Bernal et al. Relevant result in the
scientific and technical Forum, Municipal and Provincial
authoritative ranks: LeukoCIM® of the clinical trial at the
medical practice: 10 years of experience).
Giselle Pentón Rol et al. Latinoamerican award
of Pharmacology Dr. Plutarco Naranjo Vargas
in memoriam. Sponsored by the Latin American
Association of Pharmacology (LPA) and the International
Union of basic Pharmacology and clinic (IUPHAR):
effect neuroprotective and molecular mechanisms
of C-Ficocianina in experimental models of multiple
sclerosis and cerebral ischemia.
Giselle Pentón Rol et al. National Prize of Pharmacology
2011-2013, Dr. Francisco J Rodriguez Moron,
sponsored by the Cuban society of Pharmacology:
effect neuroprotective and molecular mechanisms of
the C-Ficocianina in experimental models of multiple
sclerosis and cerebral ischemia.
EVENTS AND COURSES
The logistical coordination and scientific organization of
25 CT workshops involved in 15 products and 22 annual
objectives corresponded to the members of the DCR.
Five workshops were achieved to join criteria for new CT
protocols, seven to start the implementation of the approved
CT by Cecmed, seven for scientific updating of CT in course
and six to present and discuss the results of CT concluded.
More than 150 researchers participated, and there was
a representation of more than 10 medical specialties,
with main highlight for the angeologists, gynecologists,
oncologists, virologists, dermatologists, neonatologists,
gastroenterologists and Nephrologists.
As part of the training plan arranged in the DCR, 6
specialists received professional development courses;
mainly those covered by the mention in clinical trials of
the master tendencies of contemporary biotechnology
delivered by the CIGB, as well as in English and French; for
the obtainment of PhD degrees on mathematical modeling
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YEARBOOK 2013
and its applications, three workers successfully discussed their Master’s
thesis, one in clinical trials, another in mathematics and statistics, and the
other in bioengineering.
In the DCR, teaching is also included with the participation of a group of
members as teachers of undergraduate and graduate schools, as well
as members of courts of scientific degrees both at the CIGB and the
National School of public health. Specifically the DCR has two members
of the academic Committee of the Master Degree of the CIGB, the
coordinator of the mention in CT, plus coordinators and principal teachers
of the subjects:
• Dr. Pedro López Saura. Member of the Academic Committee.
Coordinator and Main professor teaching Methodology and Practice
in CT.
• Dr. Verena Lucila Muzio González. Member of the academic
Committee. Professor of methodology and practice in CT. Main
professor of the subject Specific aspects of clinical trials in vaccines
and oncology.
• MSc. Carmen María Valenzuela Silva. Coordinator and Main
professor teaching Methods of applied statistics in biotechnology.
Also professor of the methodology and practice in CT
• MSc. Idrián García García. Coordinator and Main profesor. Principals
of pharmacology.
• Dr. Francisco Hernández Bernal. Coordinator and principal professor
teaching Principles of Epidemiology.
• Dr. Hugo Nodarse Cuní. Professor of the subject Methodology and
practice in CT.
• Dr. C. Antonieta Herrera Buch. Professor of Methodology and
practice in CT.
• Dr. Odalys Margarita González Díaz. Professor of Methodology and
practice in CT.
• Dr. Isis Belkis Yero Alós. Professor of the subject Principals of
pharmacology.
• MSc. Ángela Tuero Iglesias. Professor of the subject Methods of
applied statistics in biotechnology
Those and other colleagues of the DCR took part in six master's thesis
tribunal. The general achievement in these activities was regarded as an
institutional achievement due to maintain Master's Degree of Trends in
Modern Biotechnology (3rd. Edition) with a satisfactory level. It will start
the Fourth Edition.
Publications
Eight scientific articles were published in international journals.
SUMMARY
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YEARBOOK 2013
DIRECTION OF SAFETY AND PROTECTION
T
he Direction of Safety and Protection Safety and Protection Section of the Center
for Genetic Engineering and Biotechnology (CIGB) maintains a system of safety
management sustained in the Biosafety Program, which ensures compliance with the
regulations of the Ministry of Science, Technology and Environment, the Ministry of Labor,
the Ministry of Public Health and the international Convention on Biodiversity, the Cartagena
Protocol and conventions that regulate the use of biological, chemical and radiological
agents. It is also in charge of health-related issues in the biotechnology sector.
This group is in charge of the Strain collection laboratory, which controls biosafety and the
protection and centralization of culture collections. It also identifies, analyzes and assesses
the biological risk in the Center’s projects.
DIRECTOR
Ing. Elizabeth García Muñiz
[email protected]
Telephone: (537) 250 4556
Relevant results
• Effective regulatory activity on Biological, Chemical and Radiological Safety.
• Satisfactory results in the Defense Area.
SUMMARY
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YEARBOOK 2013
Department of Regulatory Issues and Clinical Trials
T
he Department of Regulatory Issues and Clinical Trials manages the fulfillment of
the national and international regulations in reviewing all the information for records
and clinical trials registries (CTR). It evaluates and classifies the changes in production
processes and quality control of various products of the Center for Genetic Engineering and
Biotechnology (CIGB) according to the National Center for State Control of Drugs,
Equipment and Medical Devices (CECMED) as part of the Committee of Experts. In addition,
it manages the preparation of records of all products and their processing to CECMED.
This department is responsible to the CIGB and CECMED throughout the process and
Modification Authorization Procedures for Clinical Trials, Application of Licenses in
Pharmaceutical Operations in Cuba and abroad. It makes the registry records according to
the Specific Regulations Drug Authorities of different countries , interacting with the experts
and responds to the same applicable requirements by working with the various areas of
CIGB and other centers , ensuring thus the successful performance of the Enrollment
Process , Renewal and Amendment of CIGB of the different products abroad allowing a
great commercialization of our products worldwide, with the outstanding role of the leading
product, Heberprot-P®.
Head of the department
Marisol Cruz Díaz
[email protected]
Phone: (537) 250 4188
RELEVANT RESULTS
During 2013 the main results were associated with an intense registry activity in Cuba and
abroad, allowing the obtaining of new registrations and renewals in different countries.
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YEARBOOK 2013
Registrations in Cuba
Renewal of the sanitary registration of seven products
• Hebertrans®.
• Liquid Heberon Alfa R® 3 M, 5M and 10.
• Liophilized Heberon Alfa R® 3M, 5M and 10M.
• Heberon Gamma®.
• HeberPAG® 3M and 10M.
• Heberkinasa® 750 000 IU and 1 500 000 IU.
• HeberNem®-L by the National Plant Protection
Registration Agency.
International registration
•
•
•
•
Registration renewal
Sanitary License Renewal of 7 products was obtained in 3
countries.
•
The International Registration activity is one of the main •
objectives of the Department, this ensures the use, sale •
and export of our products to all the different geographical •
areas of the world. The Registration is the initial requirement •
that precedes the use of the products in any area, including •
•
comercialization.
Obtainment of new registrations
4 Products in 5 countries
• Heberprot-P® in Peru.
Heberprot-P® in Viet Nam.
Heberitro® in Viet Nam.
Heberbiovac HB® 10 µg in Viet Nam.
Liquid Heberon® R in Venezuela.
Quimi-Hib® in Venezuela.
Heberbiovac HB® 20 µg in Tunez.
HeberFast Line® anti-transglutaminase diagnostic test in
Venezuela.
INTERNATIONAL PROJECTS
• Active and outstanding participation of the Department
Figure
Asia. 65
Eastern Europe. 42
Africa. 27
•
•
Central America. 8
Caribbean. 14
Heberprot-P® in Costa Rica
Quimi-Hib® in Marruecos.
Gavac® (Heberbiogar, in INSAI, Venezuela)
HeberFort® in Spain. Marketing Permission and
conducting tests for later bionematicide Registration in
Europe was obtained. It is the first commercial product
license of the CIGB in a European country.
South America. 64
in Different Stages of the Joint Project in 2013 for
negotiating with Wittycell with NASVAC product which
meant an income of $ 1 million for the CIGB and the
ongoing Agreements for future use of this product in new
business areas and the First World.
The Guidelines for the registration of subunit
immunogens obtained through biotechnological
processes was achieved. (Nineteenth plenary Seminar
on Harmonization of Registration and Control of
Veterinary Drugs achieved. Committee for Veterinary
Medicinal Products in America (CAMEVET).
Delivery of the final version (in English) of the
Registration form for subunit immunogens obtained
through biotechnological processes to IMV Focal Point
in Cuba, sent to all countries in America.
INSTITUTIONAL SCIENTIFIC AWARDS
• Obtainment of the marketing approval of Heberprot-P®
Current records of the Regulatory Issues of the Center for Genetic
Engineering and Biotechnology, distributed by geographic areas.
in Spain. Positive results in field trials.
• Increase in the Regulatory level of the IMPD NASVAC
dossier as balance of two satisfactory ratings by the
French Company Wittycell and the Impact of these in
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YEARBOOK 2013
•
•
•
•
•
the Schedule of Business and Development of this product with the
Company.
Renewal of the Heberprot-P® sanitary registration in Vietnam for 5
years and the obtainment of the registration in Peru and Costa Rica.
Registration request of HeberFort® and HeberNem® in Saudi Arabia
and Morocco, respectively.
Health Registration Renewal of the HeberFast Line®. in the Bolivarian
Republic of Venezuela First export of the product produced at the
Center for Genetic Engineering and Biotechnology of Sancti Spíritus
Renewal of Alpha R liquid® Heberon 3M, 5M and 10M, Heberon Alpha
R lyophilized® 3 M, 5 M and 10 M, Heberon Gamma®, HeberPAG® 3 M
and 10 M, Heberkinasa® 750 000 IU and 1500 000 IU Hebertrans®
and HeberNem®-L to the Sanitary Registration in Cuba.
Effective processing activity of national and international registration
with satisfactory compliance.
COURSES AND EVENTS MANAGED AND DELIVERED
BY THE AREA
Events
Participation in the CESFARMA Congress
• Common technical document dossier for marketing authorization in
ICH regions.
• The ASEAN Common Technical Document Dossier (ACTD) for the
registration of pharmaceuticals for human use.
Courses
• Participation in a Course with Sertere for exchanges on registration
requirements in Venezuela.
• Participation of 2 specialists as teachers in the Master's Degree of
Trends in Modern Biotechnology.
FOREIGN VISITORS
• In the months of April and November 2013 we received a visit of an
•
•
expert from the French Company Wittycell to IMPD Product Dossier
of NASVAC, managed by our department. The balance of these
two audits was very positive for both the development of the Project
as well as the ongoing negotiation with this company, and actions
in order to achieve the clinical trials and registration processes in
different parts of the world, mainly in Europe.
As a result of these audits a great step forward in the Quality and
Regulatory Compliance of this dossier was achieved, with high
impact on the negotiation with this Company.
The Department was active and positive on the working visit of
the Expert from Microguen Company from Russia, for the future
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YEARBOOK 2013
•
•
registration of the Russian Pentavalente vaccine, with the use of 2
antigens from the CIGB as Active Ingredients in this formulation: the
HBsAg and Hib.
Bioven Russia Visit: Very positive results were obtained in the
contribution in Negotiating with the Russian Company, where IMPD
Bioven Product Nasvac® was evaluated.
Visit of Dr. Numidia Corona from SERTERE, Venezuela, study and
discussion of work and quality of registration dossiers evaluated and
targeted to the Venezuelan Medicines Authority. CIGB Registrations
Recognition of Experts of that country to the Records of CIGB, as the
ones with more quality in the region.
PUBLICATIONS
Three scientific papers in which we participated as co-authors were
published in national and international magazines, three technical reports
also.
SUMMARY
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YEARBOOK 2013
CIGB in Camagüey
T
he Center for genetic Engineering and Biotechnology (CIGB) in Camagüey aims
to generate, develop, produce and commercialize biotechnology products in the
agricultural sphere and for therapy of hormone-sensitive tumors. It also contributes to the
economic, social and environmental development of our country, based on a system of
quality management that ensures the satisfaction of the customers. Work areas complete
the cycle of research-development-production and marketing
Its leader product is the recombinant vaccine against the cattle tick, commercially known
as Gavac®, of proved efficacy. It also produces and commercializes a biological product
for the control of health pesticide parasites of agricultural crops known as HeberNem®,
successfully used in houses of protected cultivation.
Its quality system guaranties the compliance with good manufacturing practices, and
constitutes an important endorsement for the products. Both the main researches and
the technological development comprise the obtainment of new bioproducts of agriculture
and veterinary use that because of its effectiveness, safety and quality are included in the
portfolio of the commercialized productions at the center.
In addition, it has a research line with medical applications. It is based on the ready
therapeutic vaccine against advanced prostate cancer, known as Heberprovac® which
recently concluded a phase II multi-center clinical study in humans with satisfactory results.
As a result of the scientific-productive work of the center, it has been achieved around
90 % of the sales revenue corresponding to exports of biotechnology products developed
in the Center.
MANAGER
Eulogio Pimentel Vázquez
[email protected]
Phone: (53-32) 26 1295
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YEARBOOK 2013
CIGB in Camagüey
Main Executives
Head of the Production
Jesús Zamora Sánchez
[email protected]
Phone: (53-32) 26 1044
Head of the Researches
Rolando Morán Valdivia
[email protected]
Phone: (53-32) 26 2798
• Scale-up and production of HeberNem® in the BioProcess
•
•
•
•
Head of the Researches
Alain Moreira Rubio
[email protected]
Phone: (53-32) 26 1044
•
Head of the Technological Development Department.
Nemecio González Fernández
[email protected]
Phone: (53-32) 26 1044
•
Main Results
• The production of more than 9 million doses of the
•
•
vaccine Gavac® and over-achievement of marketing
plans provided for this product, which is a productive
and commercial record for the institution.
Demonstration of the effectiveness of the bionematicide
HeberFort® in trials in terms of pots in Spain and its
usage record as OMDF (alternative forms of protecting
plant health) in that country.
Demonstration of safety and evidence of effectiveness
in a phase II multicenter clinical trial with the vaccine
candidate the Heberprovac® against advanced prostate
cancer.
Awards and Distinctions
Institutional Achievements
•
Cuba 10 plant of the Cuban Institute reaseraches of the
derivatives of the cane of sugar (ICIDCA).
Three years achieving satisfactory results in the
marketing of the CIGB in Camagüey.
Alternative use of EROS software to control the addition
of methanol during the fermentation of Pichia pastoris
in 300 L.
Consolidation of the business strategy for Hebernem®
and significant progress in opening markets.
Obtainment of commercial permission of HeberFort® in
Spain. Positive results in the field trials.
Progress in the negotiation of the empresa mixta in
marketing-production of agro-biotechnology products
with Shandong Lukang.
Increase in the capacity and quality standards at the
stages of formulation and filling of ImmunoGen Gavac®.
Outstanding participation in the progress of the
programme of integrated control against cattle ticks in
Cuba.
Scientific Achievements
• Determination of the plant growth promoting ability of
•
•
Tsukamurella paurometabola C-924 and characterization
of the main mechanisms involved in the process.
Lactobacillus pentosus IC1, an isolation of vegetables
fermented with probiotic activity and immunostimulating
in shrimp.
Participation in the use of glicomarkers achievement
to assess the progression of patients with prostate
cancer in the context of the phase I clinical trial with
Heberprovac®.
Events and courses received
and delivered
• A course on Molecular Biology and Functional Genomics
was managed and delivered in 54 hours in the Center
of Bioplants in Ciego de Ávila, with participants from
various provinces and national institutions.
• Productive record of ImmunoGen Gavac® for a year.
• Yielding record of API's Gavac® per processed biomass
Foreign Visitors
•
• Visit of the Ambassador of the People's Republic of
•
lots.
Recovery of equipment and solution of climates in areas
of the CIGB in Camagüey.
Quality service and maintenance of the CIGB in
Camagüey with the condition of self-financed.
China, on April 24.
• visit of executives from Lukang to discuss aspects
relating to the approval of the joint company Lukang-
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YEARBOOK 2013
CIGB in Camagüey
Heber and other inherent to technological transfers to
the Qihe plant in that country, December 27-28.
PUBLICATIONS
Four scientific articles and two technical reports were
published.
SUMMARY
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YEARBOOK 2013
CIGB in Sancti Spíritus
C
enter for Genetic Engineering and Biotechnology in Sancti Spíritus is part of the CIGB
in Havana, is located to 350 km east of the city. In its organizational structure, it counts
with different departments like: Production, Research and Development, Administration and
Service, and the Group of Assurance and Quality Control. Its main mission is to produce
biological reagents at the CIGB using control of processes, Quality controls and the release
of the main biotechnological products. In addition, it produces biological reagents for the
Center of Immunoassays (Havana, Cuba), as primary components of the Ultramicroelisa
systems for the diagnoses of the human immunodeficiency virus (HIV), the hepatitis B virus
(HBV) and C (HCV) in blood donors.
The CIGB of Sancti Spíritus counts with a manufacturing License by the Center for the
Control of Medicines, Medical equipments and Devices of Cuba, which protects the
production of biological reagents and the manufacture of the HeberFast Line® Pregnancy,
HeberFast Line® rotavirus and HeberFast Line® anti-transglutaminase, to the Department
of Public Health.
Other missions include the generation of hybrids secreting monoclonal antibodies, to
support the projects research-development by the CIGB; the development of rapid
reagents strip type- new diagnostic systems for veterinary or biomedical use; the design,
development and ratification of new immunoassays to support the stages of evaluation
of leader products at the CIGB; the genetic improvement of vet species of agricultural
interest, with projects in course aimed at the introduction of genes for the defense against
plagues and fungous illnesses in rice; the use of plants as bioreactors for the production
of proteins of pharmaceutical interest; the technology development for the production of
pre-biotic (fructooligosaccharides) and the direct intervention in the extension and use of
products of the CIGB in the province of Sancti Spíritus.
General Manager
MSc. Raúl Alfredo Armas Ramos
[email protected]
Phone: (53-41) 32 6273 ext. 102
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YEARBOOK 2013
Main Executives
Head of the Department of Research-Development
MSc. Enrique Rosendo Pérez Cruz
enrique.perez @cigb.edu.cu
Phone (53-41) 32 6273 ext. 107
Head of the Production Department
B.Sc. Omar Reinaldo Blanco Águila
[email protected]
Phone: (53-41) 32 6273 ext. 105
Chief of the Group Assurance and Quality Control
MSc. Vladimir Leal Gómez
[email protected]
Phone: (53-41) 32 6273 ext. 109
Main Results
• Renovation of the sanitary registry of HeberFast Line® antitransglutaminase, in Cuba.
• Obtainment of the sanitary registry of HeberFast Line® MaterniTest II.
• Obtainment of a production method of 1-kestosa in bioreactors of
membrane.
• Establishment of a viable technical procedure economically for the
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obtainment of the enzyme 1-SST recombinant lyophilized for the
commercial production of fructooligosaccharides (FOS).
Reestablishment of the production of similar lots of the monoclonal
antibody CBIFNα2.3, linked to the analytical system ELISA for the
quantification of interferon alpha (Heberon Alfa R®), by means of the
reclonaje and a different method of selection of clones.
Development of new monoclonal antibodies to support the research
and development of new products at the CIGB like the vaccine
candidates against Bordetella pertussis, HCV and the sea louse.
Renewal of the sanitary registry in the Republic of Venezuela of
HeberFast Line anti-transglutaminase. The first exportation of the
diagnosticians produced by the CIGB of Sancti Spíritus.
Establishment of a purification procedure, with more than 85 % of
purity, of the streptokinase expressed in chloroplasts of tobacco
based on operations of filtration, precipitation with sulphate of
ammonium and chromatography of ionic exchange.
Obtainment of a thermosetting safety, biocatalyst producing inverted
sugar, which can be used to industrial scale to obtain invested sugar
in high temperatures and concentration of saccharose.
Development and validation of an inquiry immunoassay and another
confirmatory to evaluate immunogenicity not desired of Heberprot-P®,
a fundamental requirement to the marketing of the product to the first
world.
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• Obtainment of rice lines genetically modified, free of the selection
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marker gene, expressing the NmDef02 resistant gene to diseases (3
lines) and the Cry1Fa resistant gene to insects (7 lines).
Overachievement of the production plan at 124.4 % of biological
reagents and biomedical diagnosticians of the CIGB in Sancti
Spíritus with the consequent economic contribution. This guaranteed
a positive balance in the economic results of the Center, for the
second time in years.
Establishment of an active inquiry system through operatives to
the prevention, specialized attention and precocious for the use
of Heberprot-P® in patients with diabetic foot ulcers in the primary
attention of health.
Awards and Distinctions
• Recognition as Center emphasized in the activity of the science and
the technological innovation in the province, granted by the delegation
of the Department of Science Technology and Environment (Citma)
of Sancti Spíritus.
Events y courses
• The IIIrd Provincial Workshop of the of integral attention Program to
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patients with diabetic foot ulcer (PAIPUD) and Heberprot-P®, Sancti
Spíritus, October, 2013.
A diploma on integral Handling to patients with diabetic foot ulcers in
the province of Sancti Spíritus.
A diploma on surgical Procedures in patients with diabetic foot ulcers,
together with the Angiology room of the Provincial Hospital Camilo
Cienfuegos, Sancti Spíritus.
PublicationS
Six scientific articles and a patent application were published.
SUMMARY
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PUBLICATIONS
ARTICLES
PUBLISHED IN 2013
• Acosta J, Montero V, Carpio Y, Velázquez J, Garay HE, Reyes O, et al. Cloning
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and functional characterization of three novel antimicrobial peptides from tilapia
(Oreochromis niloticus). Aquaculture. 2013;372–375:9-18.
Aguilar JC. Highlights from Barcelona 2012 International Liver Congress and EASL
meeting. Biotecnología Aplicada. 2013;30(2):145-8.
Aguilera A, Bermudez Y, Martínez E, Marrero MA, Muñoz L, Páez R, et al. Formulation
development of a recombinant Streptokinase suppository for hemorrhoids treatment.
Biotecnología Aplicada. 2013;30(3):182-6.
Alvarez T, Ferro W, Geada D, Medina Y, Montero J, Muñoz L, et al. Detection
of an immunologically-relevant epitope in the Heberbiovac® vaccine active
pharmaceutical ingredient employing CB.Hep-4 monoclonal antibody. Lat Am J
Pharm. 2013;32(9):1280-7.
Aragón H, González M, Valdés R, Álvarez T, Brown E, Rodríguez Y, et al. Replacement
of serum supplemented medium for CB.Hep-1 hybridoma cell freezing and monoclonal
antibody production. Biotecnología Aplicada. 2013;30(1):57-62.
Bacardí D, Cosme K, Aldana L, Merino N, Suárez J, Mosqueda O, et al. Preclinical
safety testing of the Quimi-Hib® vaccine adjuvanted with aluminum phosphate during
product development. Biotecnología Aplicada. 2013;30(2):118-24.
Barbará E, Saurez G, Cabal Mirabal C, González Dalmau E, editors. Tool of segmentation
and 3D reconstruction of MRI to quantify cranial tumor activityImage. XVIII Symposium
of Image Signal Processing, and Artificial Vision (STSIVA) 2013; Bogotá, Colombia.
Barbera A, Lorenzo N, Garrido G, Mazola Y, Falcon V, Torres AM, et al. APL-1, an altered
peptide ligand derived from human heat-shock protein 60, selectively induces apoptosis
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in activated CD4+ CD25+ T cells from peripheral
blood of rheumatoid arthritis patients. International
Immunopharmacology. 2013;17(4):1075-83.
Bello-Rivero I, Garcia-Vega Y, Valenzuela-Silva C,
Bello-Alvarez C, Vázquez-Blomquist D, Lopez-Saura
P. Development of a new formulation of interferons
(HEBERPAG) for BCC treatment. Journal of Cancer
Research and Therapeutics. 2013;1(10):235-43.
Berlanga J, Fernandez JI, Lopez E, Lopez PA, del Rio
A, Valenzuela C, et al. Heberprot-P®: a novel product for
treating advanced diabetic foot ulcer. MEDICC Review.
2013;15(1):11-5.
Berlanga-Acosta J, López-Saura P, Guillen-Pérez I,
Guillen-Nieto G, Acevedo-Castro B, Herrera-Martínez L.
Type 2 Diabetes Mellitus (T2DM): Biological overview
from pathways to organelles and its translation toward
a torpid wound healing process. J Diabetes Metab
2013;4:7.
Berlanga-Acosta J, Mendoza-Mari Y, Martinez MD,
Valdes-Perez C, Ojalvo AG, Armstrong DG. Expression
of cell proliferation cycle negative regulators in fibroblasts
of an ischemic diabetic foot ulcer. A clinical case report.
International wound journal. 2013;10(2):232-6.
Berlanga-Acosta J, Schultz GS, Lopez-Mola E, GuillenNieto G, Garcia-Siverio M, Herrera-Martinez L. Glucose
toxic effects on granulation tissue productive cells:
the diabetics' impaired healing. BioMed Research
International. 2013;2013:256043.
Betancourt LH, De Bock PJ, Staes A, Timmerman E,
Perez-Riverol Y, Sanchez A, et al. SCX charge state
selective separation of tryptic peptides combined with
2D-RP-HPLC allows for detailed proteome mapping.
Journal of Proteomics. 2013;91:164-71.
Boraschi D, Penton-Rol G. Perspectives in
immunopharmacology: The future of immunosuppression.
Immunology Letters. 2013.
Borges D, Perez-Riverol Y, Nogueira FC, Domont GB,
Noda J, da Veiga Leprevost F, et al. Effectively addressing
complex proteomic search spaces with peptide spectrum
matching. Bioinformatics. 2013;29(10):1343-4.
Cabal C, Darias D, González E, Musacchio A.
Theranostics and Molecular Imaging: new concepts
and technologies for drug development. Biotecnología
Aplicada. 2013;30(3):172-7.
Cabal Mirabal C. Ciencia y sociedad: ¿qué pasará en
los próximos 20 años? Juventud Técnica. 2013;2013.
Cabal Mirabal C, Gonzalez Dalmau E. Las imágenes
moleculares y las micro imágenes. Nuevos retos a la
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bioingeniería. V Latin American Congress on Biomedical
Engineering CLAIB IFMBE Proceedings 2013;33:531-4.
Cabrales A, Gil J, Fernandez E, Valenzuela C, Hernandez
F, Garcia I, et al. Pharmacokinetic study of Growth
Hormone-Releasing Peptide 6 (GHRP-6) in nine male
healthy volunteers. European Journal of Pharmaceutical
Sciences. 2013;48(1-2):40-6.
Camacho H, Fernandez ME, Guillen IA, Perez L,
Fernandez JR, Tuero AD, et al. Analysis of cell
proliferation and gene expression profiles in Epidermal
Growth Factor-treated tumor cell lines. Minerva
Biotecnol. 2013;25(1):43-54.
Carpio Y, Garcia C, Pons T, Haussmann D, RodriguezRamos T, Basabe L, et al. Akirins in sea lice: first
steps towards a deeper understanding. Experimental
Parasitology. 2013;135(2):188-99.
Díaz B, Heynngnezz L, Beldarraín A, Iser Y, Fernández
A, Díaz J, et al. Cromatografía de adsorción para purificar
el disacárido espaciador del ingrediente farmacéutico
activo de la vacuna QuimiHib®. VacciMonitor.
2013;22(1):29-34.
Domínguez MC, Lorenzo N, Barberá A, Padrón G,
Torres AM, Hernández MV, et al. Therapeutic effect
of two altered peptide ligands derived from the
human heat shock protein 60 in experimental models
of rheumatoid arthritis. Biotecnología Aplicada.
2013;30(2):153-6.
Fernandez E, Toledo JR, Mendez L, Gonzalez N, Parra
F, Martin-Alonso JM, et al. Conformational and thermal
stability improvements for the large-scale production
of yeast-derived rabbit hemorrhagic disease viruslike particles as multipurpose vaccine. PloS One.
2013;8(2):e56417.
Fernandez Masso JR, Oliva Arguelles B, Tejeda Y,
Astrada S, Garay H, Reyes O, et al. The Antitumor
Peptide CIGB-552 Increases COMMD1 and Inhibits
Growth of Human Lung Cancer Cells. Journal of Amino
Acids. 2013;2013:251398.
Fernandez Nuñez EG, Valdes Veliz R, Vale da Costa
BL, Goncalves de Rezende A, Tonso A. Using statistical
tools for improving bioprocesses. Asian J Biotechnol.
2013;5(1):1-20.
Fernández-Ortega C, Casillas D, Dubed M, Navea L,
Ramírez A, López L, et al. Leukocyte extract modulates
cellular factors involved in HIV infection. Sexually
Transmitted Infect. 2013;89:45-7.
Fleitas Díaz M, Rodríguez Reyes O, Benítez Pardillo
T, Mena Campos J, Mesa L. Evaluación de dosis de
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HeberNem para el control de Meloidogyne incognita
Chitwood en condiciones de cultivos protegidos. Centro
Agrícola. 2013;40(1):57-62.
Garcia Del Barco-Herrera D, Martinez NS, CoroAntich RM, Machado JM, Alba JS, Salgueiro SR, et
al. Epidermal growth factor and growth hormonereleasing peptide-6: combined therapeutic approach
in experimental stroke. Restorative Neurology and
Neuroscience. 2013;31(2):213-23.
García O, Nelson E, Godinez L, Cabrera D, Sosa R,
Martínez M, et al. Improvements in an ion-exchange
chromatography procedure to increase recovery and
biological activity of erythropoietin preparations for
pharmaceutical use. Lat Am J Pharm. 2013;32(6):852-9.
Garcia-Diaz D, Rodriguez I, Santisteban Y, Marquez
G, Terrero Y, Brown E, et al. Th2-Th1 shift with the
multiantigenic formulation TERAVAC-HIV-1 in Balb/c
mice. Immunology Letters. 2013;149(1-2):77-84.
Gomez L, Padilla S, Fuentes A, Ruiz Y, Gonzalez
T, Somoza M, et al. Assessment of Two Transgenic
Tobacco Plant Varieties for the HBsAg-Specific
Plantibody Production J Agron. 2013;12:11-9.
Gonzalez Dalmau E, Cabal Mirabal C, editors. Criterios
Cuantitativos de Selección de la Bobina Receptora de
Radiofrecuencia para Estudiar Animales Pequeños en
Equipos Clínicos de Resonancia Magnética. V Latin
American Congress on Biomedical Engineering CLAIB
2011 May 16-21, 2011, Habana, Cuba; 2013 2013/01/01:
Springer Berlin Heidelberg.
González-Fernández N, Ramos-Sánchez L, NarciandiDíaz E, Mayo-Abad O, Zamora-Sánchez J. Tecnología
de fermentación del agente biológico activo del
bionematicida HeberNem®. Rev Cubana Quím.
2013;25(1):55-65.
Granadillo M, Batte A, Lugo VM, Musacchio A, BequetRomero M, Betancourt L, et al. Expression, purification
and characterization of a recombinant fusion protein
based on the human papillomavirus-16 E7 antigen.
SpringerPlus. 2013;2(1):12.
Guillen G, Aguilar JC, Dueñas S, Hermida L, Iglesias
E, Penton E, et al. Virus-like particles as nanovaccine
candidates. Adv Nat Sci Nanosci Nanotechnol.
2013;4(1):4.
Guillén I, Berlanga J, Camacho H, Fernández-de-Cossío
ME, Pérez L, Novoa LI, et al. Molecular mechanisms
involved in the inhibition of tumor cells proliferation
exposed to elevated concentrations of the epidermal
growth factor. Biotecnología Aplicada. 2013;30(3):223-7.
• Hernández A, López A, Ceballo Y, Rosabal L, Rosabal
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Y, Tiel K, et al. High-level production and aggregation of
hepatitis B surface antigen in transgenic tobacco seeds.
Hernández D, Cabrera L, Valdés R, Morán I, Téllez P,
Ramos Y, et al. Bacillus thuringiensis Vip3Aa1 Expression
and Purification from E. coli to be Determined in Seeds
and Leaves of Genetically Modified Corn Plants. J
Agron. 2013;12(4):153-67.
Hernandez-Bernal F, Valenzuela-Silva CM, QuinteroTabio L, Castellanos-Sierra G, Monterrey-Cao D,
Aguilera-Barreto A, et al. Recombinant streptokinase
suppositories in the treatment of acute haemorrhoidal
disease. Multicentre randomized double-blind placebocontrolled trial (THERESA-2). Colorectal disease: the
official journal of the Association of Coloproctology of
Great Britain and Ireland. 2013;15(11):1423-8.
Herrera L. Biotecnología Aplicada: 30 years publishing
science. Biotecnología Aplicada. 2013;30(1):ii.
Iglesias E. Is there any room for therapeutic vaccination
against the HIV-1/AIDS? Human Vaccines &
Immunotherapeutics. 2013;9(7):1539-44.
Lamdan H, Gavilondo JV, Munoz Y, Pupo A, Huerta V,
Musacchio A, et al. Affinity maturation and fine functional
mapping of an antibody fragment against a novel
neutralizing epitope on human vascular endothelial
growth factor. Molecular Biosystems. 2013;9(8):2097-106.
Lemos G, Guillén I, Fernández JR, Díaz T, Colarte AB,
Fernández de Cossío ME. Expression and purification
of a full-length recombinant NS1 protein from a dengue
2 serotype viral isolate. Biotecnología Aplicada.
2013;30(3):187-93.
Leprevost FV, Lima DB, Crestani J, Perez-Riverol Y,
Zanchin N, Barbosa VC, et al. Pinpointing differentially
expressed domains in complex protein mixtures with the
cloud service of PatternLab for Proteomics. Journal of
Proteomics. 2013;89:179-82.
Limonta M, Krajnc NL, Vidic U, Zumalacárregui L.
Simulation for the recovery of plasmid for a DNA vaccine.
Biochem Eng J. 2013;80:14-8.
López-Saura PA, Yera-Alos IB, Valenzuela-Silva C,
González-Díaz O, del Río-Martín A, Berlanga-Acosta
J, et al. Medical practice confirms clinical trial results of
the use of intralesional human recombinant Epidermal
Growth Factor in advanced diabetic foot ulcers. Adv
Pharmacoepidem Drug Safety 2013;2:128.
Lugo JM, Carpio Y, Morales R, Rodriguez-Ramos
T, Ramos L, Estrada MP. First report of the pituitary
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adenylate cyclase activating polypeptide (PACAP) in
crustaceans: conservation of its functions as growth
promoting factor and immunomodulator in the white
shrimp Litopenaeus vannamei. Fish & Shellfish
Immunology. 2013;35(6):1788-96.
Ma JK, Christou P, Chikwamba R, Haydon H, Paul M,
Ferrer MP, et al. Realising the value of plant molecular
pharming to benefit the poor in developing countries
and emerging economies. Plant Biotechnology Journal.
2013;11(9):1029-33.
Marcos E, Gil L, Lazo L, Izquierdo A, Brown E, Suzarte
E, et al. Purified and highly aggregated chimeric
protein DIIIC-2 induces a functional immune response
in mice against dengue 2 virus. Archives of Virology.
2013;158(1):225-30.
Marín Bruzos M, Mena Campos J, Chaveli Chávez P,
Morán Valdivia R, Pimentel Vázquez E. Interaction
among Tsukamurella paurometabola C-924 and
Rhizobium leguminosarum biovar phaseoli CFH in bean
plants. Acta Agronoómica. 2013;62(1):52-8.
Marín M, Wong I, García G, Morán R, Basulto R, Pimentel
P, et al. Actividad antagónica in vitro de Tsukamurella
paurometabola C-924 frente a fitopatógenos. Rev
Protección Veg. 2013;28(2):132-7.
Marín M, Wong I, Wong J, Morán R, Pimentel E,
Sánchez I, et al. Zea mays L. plant growth promotion
by Tsukamurella paurometabola strain C-924.
Marin-Prida J, Pavon-Fuentes N, Llopiz-Arzuaga A,
Fernandez-Masso JR, Delgado-Roche L, Mendoza-Mari
Y, et al. Phycocyanobilin promotes PC12 cell survival
and modulates immune and inflammatory genes and
oxidative stress markers in acute cerebral hypoperfusion
in rats. Toxicology and Applied Pharmacology.
2013;272(1):49-60.
Martínez R, Estrada MP, Ubieta K, Herrera F, Forellat A,
Gil L, et al. Biological activity in vitro and in vivo of an in
silico designed secretagogue peptide to be used in fish.
Medell M, Hart M, Duquesne A, Espinosa F, Valdes R.
Nosocomial ventilator-associated pneumonia in Cuban
intensive care units: bacterial species and antibiotic
resistance. MEDICC Review. 2013;15(2):26-9.
Mendoza-Marí Y, Valdés-Pérez C, Rodríguez-Corrales
E, Suárez-Alba J, García-Ojalvo A, García del Barco
Herrera D, et al. Histological and transcriptional
expression differences between diabetic foot and
pressure ulcers. J Diabetes Metab 2013;4:296.
• Menendez C, Martinez D, Trujillo LE, Mazola Y, Gonzalez
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E, Perez ER, et al. Constitutive high-level expression
of a codon-optimized beta-fructosidase gene from
the hyperthermophile Thermotoga maritima in Pichia
pastoris. Applied Microbiology and Biotechnology.
2013;97(3):1201-12.
Miyares M, González J, Torres D, Muñoz L, Pestana Y,
Padrón S, et al. Validación de la técnica de determinación
de proteínas totales por el método microcoomassie a
doble longitud de onda para la muestra de producto
terminado del antígeno de la nucleocápsida del virus de
la hepatitis C. VacciMonitor. 2013;22(2):14-8.
Morán Y, Chacón O, Córdoba-Sellés MC, DomínguezLarrinaga R, Herrera L, Borrás-Hidalgo O. Identification
and Molecular Characterization of Nocardia sp. as a
New Causal Agent of Tobacco False Broomrape. J
Phytopathol. 2013;161(2):86-91.
Muñoz L, Font M, García G, Pérez N, Torres D, Álvarez
D, et al. Validation and application of a sensitive ELISA
to quantify rec-GCSF for preventing cross-contamination
in recombinant proteins produced in a multi-product
facility. Lat Am J Pharm. 2013;32(3):400-8.
Nodarse-Cuní H, J.A. M-P, Gutierrez-Pérez Y,
Valenzuela-Silva CM, Lazo-Diago OC, GutierrezAlvarez C, et al. Epidermal growth factor enemas for
induction of remission in left-sided ulcerative colitis. Rev
Cubana Farm. 2013;47(1):67-76.
Nodarse-Cuní H, Más-Paez A, Gutierrez-Pérez Y,
Valenzuela-Silva CM, Lazo-Diago OC, Gutierrez-Alvarez
C, et al. Enemas de factor de crecimiento epidérmico
para inducir la remisión de la colitis ulcerosa izquierda.
Rev Cubana Farm. 2013;47(1):67-76.
Ojeda Y, Heynngnezz L, García J, Valdés Y, González
CA, Rodríguez N, et al. Aplicación del análisis de riesgo
en la preparación de soluciones para producción de
Quimi-Hib® VacciMonitor. 2013;22(2):19-23.
Oliva BM, Fernández JR, Tejeda Y, Astrada S, Garay HE,
Reyes O, et al. P02.05 Pharmacological stabilization of
COMMD1 mediated by CIGB-552 regulates NF-KB/
oxidative stress signaling and inhibits lung cancer cells
growth. Ann Oncol. 2013;24(suppl 1):i21.
Pérez Bernal M, Abreu Remedios D, Valdivia Pérez O,
Delgado Rigo M, Armas Ramos R. Effective β-lactam
antibiotics for Agrobacterium tumefaciens suppression
in indica rice calli. Rev Colombiana Biotecnol.
2013;15(2):108-17.
Perez-Riverol Y, Hermjakob H, Kohlbacher O, Martens
L, Creasy D, Cox J, et al. Computational proteomics
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pitfalls and challenges: HavanaBioinfo 2012 workshop
report. Journal of Proteomics. 2013;87:134-8.
Perez-Riverol Y, Sanchez A, Noda J, Borges D, Carvalho
PC, Wang R, et al. HI-bone: a scoring system for
identifying phenylisothiocyanate-derivatized peptides
based on precursor mass and high intensity fragment
ions. Analytical Chemistry. 2013;85(7):3515-20.
Raíces M. II International congress addressed to the
integral care of diabetic foot ulcer patient with the use of
Heberprot-P 2012: healthy doorways wide-open to the
2014 edition. Biotecnología Aplicada. 2013;30(1):63-4.
Rodríguez EG. La revisión editorial por pares: rechazo
del manuscrito, deficiencias del proceso de revisión,
sistemas para su gestión y uso como indicador científico.
RCICS (Acimed). 2013;24(3):313-29.
Rodríguez EG. La revisión editorial por pares: Roles y
procesos. RCICS (Acimed). 2013;24(2):160-75.
Rodríguez EG, Valdés RA, Ferrer ME, Peña DA.
A uniform digital identifier for scientific articles from
manuscript through citation. Biotecnología Aplicada.
2013;30(2):142-4.
Rodríguez M, González MC, Cristo E, Oliva O, Pujol M,
Borrás-Hidalgo O. Identification of genes with altered
expression levels in contrasting rice cultivars exposed
to salt stress treatments. Biotecnología Aplicada.
2013;30(3):178-81.
Rodriguez M, Perez L, Gavilondo JV, Garrido G, BequetRomero M, Hernandez I, et al. Comparative in vitro
and experimental in vivo studies of the anti-epidermal
growth factor receptor antibody nimotuzumab and its
aglycosylated form produced in transgenic tobacco
plants. Plant Biotechnology Journal. 2013;11(1):53-65.
Rodríguez M, Pujol M, Pérez L, Gavilondo JV, Garrido
G, Ayala M, et al. Transgenic plants of Nicotiana
tabacum L. express aglycosylated monoclonal antibody
with antitumor activity. Biotecnología Aplicada.
2013;30(2):157-61.
Rodríguez O, Rodríguez JA, Pérez ER, Santos B, Cortada
A, Trujillo LE, et al. Evaluación de una fermentación
láctica en presencia de un sirope prebiótico. Cienc
Tecnol Alim. 2013;23(1):14-9.
Rodríguez-Alonso I, García-Díaz D, Santisteban YC,
García D, Soria Y, Brown E, et al. Report of the Conference
on Immunological Mechanisms of Vaccination;
December 13-18, 2012, Ottawa, Canada. Biotecnología
Aplicada. 2013;30(3):216-21.
Rodríguez-Mallon A, Bechara G, Zacarias R, BenavidesOrtiz E, Soto-Rivas J, Gómez-Ramírez A, et al. Inhibition
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of Ehrlichia canisand Babesia canis transmission
among ticks fed together on dogs vaccinated with Bm86
antigen. Open J Anim Sci. 2013;3:24-32.
Ruiz A, Hernandez Y, Cabal C, Gonzalez E, VeintemillasVerdaguer S, Martinez E, et al. Biodistribution and
pharmacokinetics of uniform magnetite nanoparticles
chemically modified with polyethylene glycol. Nanoscale.
2013;5(23):11400-8.
Saez V, Ramon JA, Caballero L, Aldana R, Cruz E,
Peniche C, et al. Extraction of PLGA-microencapsulated
proteins using a two-immiscible liquid phases system
containing surfactants. Pharmaceutical Research.
2013;30(2):606-15.
Sánchez García JC, Hernández Caso N, Hernández
González R, Musacchio Lasa A, Llopiz Arzuaga A,
Santana Millan H, et al. Definition of a security value
determined by Limulus Amebocyte Lysate assay
targeting the recombinant Human Epidermal Growth
Factor. BioPharm Int. 2013;26(10):44-52.
Santana H, Gonzalez Y, Campana PT, Noda J, Amarantes
O, Itri R, et al. Screening for stability and compatibility
conditions of recombinant human epidermal growth
factor for parenteral formulation: effect of pH, buffers,
and excipients. International Journal of Pharmaceutics.
2013;452(1-2):52-62.
Santos A, Reyes O, Cabrales A, Rodríguez Y, Geronimo
H, Garay HE, et al. Identification of the first antagonist
peptide that inhibits biological effects of interleukin-15.
Silva Y, Portieles R, Pujol M, Terauchi R, Matsumura
H, Serrano M, et al. Expression of a microbial serine
proteinase inhibitor gene enhances the tobacco defense
against oomycete pathogens. Physiol Mol Plant Pathol.
2013;84:99-106.
Soriano-García
JL,
López-Díaz
A,
SolaresAsteasuainzarra M, Baladrón-Castrillo I, BatistaAlbuerne N, García-García I, et al. Pharmacological
and safety evaluation of CIGB-300, a casein kinase 2
inhibitor peptide, administered intralesionally to patients
with cervical cancer stage IB2/II. Journal of Cancer
Research and Therapeutics. 2013;1:163.
Soto N, Ferreira A, Delgado C, Enríquez G. In vitro
regeneration of soybean plants of the Cuban Incasoy-36
variety. Biotecnología Aplicada. 2013;30(1):34-8.
Tamayo A, Geada D, Fernández EG, Dorta L,
Padilla S, Cecilia D, et al. Demonstration of
affinity chromatography stability used to purify a
monoclonal antibody employed as inmunoreagent in
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hepatitis B vaccine manufacturing. Lat Am J Pharm.
2013;32(4):543-9.
Vaillant JD, Fraga A, Diaz MT, Mallok A, Viebahn-Hansler
R, Fahmy Z, et al. Ozone oxidative postconditioning
ameliorates joint damage and decreases proinflammatory cytokine levels and oxidative stress in
PG/PS-induced arthritis in rats. European Journal of
Pharmacology. 2013;714(1-3):318-24.
Valdes I, Gil L, Castro J, Odoyo D, Hitler R, Munene
E, et al. Olive baboons: a non-human primate model
for testing dengue virus type 2 replication. International
Journal of Infectious Diseases. 2013;17(12):e1176-81.
Valdés I, Hermida L, Gil L, Lazo L, Guillén G, Guzmán
MG, et al. Proof of concept in non-human primates of the
heterologous prime-boost strategy combining dengue-2
virus and recombinant proteins including domain III
of the viral envelope protein. Biotecnología Aplicada.
2013;30(2):150-2.
Valenzuela-Silva CM, Tuero-Iglesias AD, Garcia-Iglesias
E, Gonzalez-Diaz O, Del Rio-Martin A, Yera Alos IB,
et al. Granulation response and partial wound closure
predict healing in clinical trials on advanced diabetes
foot ulcers treated with recombinant human epidermal
growth factor. Diabetes Care. 2013;36(2):210-5.
Valiente J, García del Barco D, Guillén G, Santana
H, Altruda F, Tarone G, et al. Cardiotropic effect of
GHRP-6: in vivo characterization by echocardiography.
Vera R, Perez-Riverol Y, Perez S, Ligeti B, KerteszFarkas A, Pongor S. JBioWH: an open-source Java
framework for bioinformatics data integration. Database.
2013;2013:bat051.
Vizcaino JA, Cote RG, Csordas A, Dianes JA, Fabregat
A, Foster JM, et al. The Proteomics Identifications
(PRIDE) database and associated tools: status in 2013.
Nucleic Acids Research. 2013;41(D1):D1063-9.
Yera-Alos IB, Alonso-Carbonell L, Valenzuela-Silva
CM, Tuero-Iglesias AD, Moreira-Martinez M, MarreroRodriguez I, et al. Active post-marketing surveillance of
the intralesional administration of human recombinant
epidermal growth factor in diabetic foot ulcers. BMC
Pharmacology & Toxicology. 2013;14:44.
SUBMITTED IN 2013 AND FURTHER PUBLISHED
• Aguilera A, Muñoz L, Bermúdez Y, Arias D, Martínez
Y, García G, et al. Validation of a chromogenic
substrate method for biological activity quantification of
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Streptokinase extracted from suppositories intended to
treat patients with hemorrhoids. Lat Am J Pharm. 2013;
Forthcoming.
Alvarez A, Tambara Y, Alvarez F, Besada V,
Gonzalez LJ. Calificación de un cromatógrafo de
gases Agilent Technologies 7890A utilizado en la
industria biotecnológica. Rev Cubana Farm. 2014;48
(Forthcoming).
Amador-Canizares Y, Martinez-Donato G, AlvarezLajonchere L, Vasallo C, Dausa M, Aguilar-Noriega D, et
al. HCV-specific immune responses induced by CIGB230 in combination with IFN-alpha plus ribavirin. World
Journal of Gastroenterology. 2014;20(1):148-62.
Audain E, Sanchez A, Vizcaino JA, Perez-Riverol
Y. A survey of molecular descriptors used in mass
spectrometry based proteomics. Current Topics in
Medicinal Chemistry. 2014;14(3):388-97.
Bello C, Vázquez-Blomquist D, Miranda J, García Y,
Novoa LI, Palenzuela D, et al. Regulation by IFN-α/
IFN-γ Co-Formulation (HerberPAG®) of Genes Involved
in Interferon-STAT-Pathways and Apoptosis in U87MG.
Current Topics in Medicinal Chemistry. 2014;14(3):351-8.
Dalmau ER, Cabal Mirabal C, Martinez GS, Davila
AL, Suarez JC, Cabanas Armada R, et al. Brain tumor
response to nimotuzumab treatment evaluated on
magnetic resonance imaging. Pediatrics International.
2014;56(1):43-6.
Gavilondo JV, Hernandez-Bernal F, Ayala-Avila M, de
la Torre AV, de la Torre J, Morera-Diaz Y, et al. Specific
active immunotherapy with a VEGF vaccine in patients
with advanced solid tumors. Results of the CENTAURO
antigen dose escalation phase I clinical trial. Vaccine. 2014.
Hernandez Bernal F, Lazo Diago OC, Bermudez Badell
C. Caracterización epidemiológica de la enfermedad
hemorroidal aguda en hospitales seleccionados. Rev
Cubana Hig Epidemiol. 2014;Forthcoming.
Hernández-Bernal F, Castellanos-Sierra G, ValenzuelaSilva CM, Catasús-Álvarez KM, Valle-Cabrera R,
Aguilera-Barreto A, et al. Recombinant streptokinase vs
phenylephrine-based suppositories in acute hemorrhoids,
randomized, controlled trial (THERESA-3). World Journal
of Gastroenterology. 2014;20(6):1594-601.
Llinas SG, Caballero AJ, Penalver JC, Valdes R. Platelet
serotonin concentration and clinical status in alcohol
withdrawal syndrome, preliminary results. MEDICC
Review. 2014;16(1):37-42.
Martinez D, Cutino-Avila B, Perez ER, Menendez C,
Hernandez L, Del Monte-Martinez A. A thermostable
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exo-beta-fructosidase immobilised through rational
design. Food Chemistry. 2014;145:826-31.
Perez-Riverol Y, Wang R, Hermjakob H, Muller M,
Vesada V, Vizcaino JA. Open source libraries and
frameworks for mass spectrometry based proteomics: A
developer's perspective. Biochimica et Biophysica Acta.
2014;1844(1 Pt A):63-76.
Ramos Gomez Y, Gallien S, Huerta V, van Oostrum
J, Domon B, Gonzalez LJ. Characterization of Protein
Complexes using Targeted Proteomics. Current Topics
in Medicinal Chemistry. 2014;14(3):344-50.
Villavicencio-Diaz TN, Rodriguez-Ulloa A, Guirola-Cruz
O, Perez-Riverol Y. Bioinformatics tools for the functional
interpretation of quantitative proteomics results. Current
Topics in Medicinal Chemistry. 2014;14(3):435-49.
• Amado León González, Héctor Lucas Díaz Balaguer,
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TECHNICAL REPORTS
• Alberto Leyva, Joel Ríos, Iriac Bisquet, Manuel Montané,
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Michel Nogueira, Yusniel Puig, Tania de la Cruz, Rebeca
Vizcaíno, Nancy Noa, Dalgys E. Rodríguez, Alexander
Hernández. Evaluación del funcionamiento de los filtros
de carbón activado del sistema de agua purificada de la
planta de producción.
Amado León González, Frank García Llanes, Héctor
Lucas Díaz Balaguer, José Roberto Motolongo
Viquillón, Francisco Joel Hernández Peñalver, Pedro
Pérez Llanes, Reynaldo Cardoso Infante, Vivian Pujol
García, José Blas Tejera, Lisbiel Hernández Capestany,
Eulalia Maritza Rivera, Nelvis Lamar Herrera, Cilia
Maria Mederos Robaina, Lisbett Melo Acosta, Ramón
Trujillo Marici, Rebeca Bouyón Albarrán y Rachel
Berrios Martínez. Evaluación del tiempo de vencimiento
de las placas con medio de cultivo sólido inoculadas y
sin inocular empleadas en la producción de FCE-hr para
uso parenteral.
Amado León González, Frank García Llanes, Héctor
Lucas Díaz Balaguer, José Roberto Motolongo Viquillón,
Francisco Joel Hernández Peñalver, Pedro Pérez
Llanes, Reynaldo Cardoso Infante, Vivian Pujol García,
José Blas Tejera, Lisbiel Hernández Capestany, Eulalia
Maritza Rivera, Christian Moreno León, Bárbara Pérez
Montoto, Meyli Sánchez Pardo, Nelvis Lamar Herrera,
Cilia Maria Mederos Robaina, Lisbett Melo Acosta,
Ramón Trujillo Marici, Rebeca Bouyón Albarrán y Rachel
Berrios Martínez. Empleo del pantotenato de calcio
monohidratado (NP 590) en la etapa de multiplicación del
proceso de fermentación de FCE-hr para uso parenteral.
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Pedro Pérez Llanes, Frank García Llanes, José Roberto
Motolongo Viquillón, Francisco Joel Hernández Peñalver,
Reynaldo Cardoso Infante, Vivian Pujol García, Ramón
Trujillo Marici y Rebeca Bouyón Albarrán. Evaluación
del cambio de filtros de aire del fermentador de 75 L
ubicado en Planta 4.
Amado León González, José Blas Tejera, Frank García
Llanes, Lisbiel Hernández Capestany, Héctor Lucas
Díaz Balaguer, José Roberto Motolongo Viquillón,
Francisco Joel Hernández Peñalver, Pedro Pérez
Llanes, Reynaldo Cardoso Infante, Vivian Pujol García,
Eulalia Maritza Rivera, Christian Moreno León, Bárbara
Pérez, Meyli Sánchez Pardo, Nelvis Lamar Herrera,
Cilia Maria Mederos Robaina, Lisbett Melo Acosta,
Ramón Trujillo Marici, Rebeca Bouyón Albarrán y Rachel
Berrios Martínez. Utilización del pantotenato de calcio
monohidratado (NP 590) en la etapa de multiplicación del
proceso de fermentación de FCE-hr para uso parenteral.
Arturo González Juiz, Francisco Machado Ramírez,
Criseys Peña Benítez, Angela Maria Leyva Hernández,
Lilia Luisa Pérez Suárez, Lourdes Costa Anguiano,
Makis Torres Toledo, Lázaro Heynngnezz Pérez, Jorge
Luis Vega, Eduardo Martínez Díaz, Yair Quiñones
Maya. Confección, aprobación y firma del CONTRATO
de Transferencia de Tecnología para la Formulación,
Llenado y Envase de la Vacuna Quimi-Hib® entre Heber
Biotec S. A. y MICROGEN, Rusia.
Arturo González Juiz, Francisco Machado Ramírez,
Eduardo Martínez Díaz, Criseys Peña Benítez, Angela
Maria Leyva Hernandez, Lilia Luisa Pérez Suárez,
Dinorah Torres Idaody, Rolando Paez Meireles, Ileana
Rosales Torres, Lourdes Costa Anguiano, Oscar Cruz
Gutierrez, Jorge Luis Vega, Raimundo Ubieta Gómez,
Emilio Narciandi Díaz, Hugo Nodarse Cuni, Gudelia
Perez Monras y Yair Quiñones Maya. Confección,
aprobación y firma del Contrato de Transferencia de
Tecnología para la Formulación, Llenado y Envase del
Interferón Alfa 2b pegilado entre Heber Biotec S. A. y
PHARMSTANDARD, Rusia.
Belkis Lérida Abreu Piñeiro, Yohanix López García,
Daniel González Aguilar, Oscar Cruz Gutiérrez, Jacssel
Zaldívar Fernández, Ronald de Oro Jorge, Aleydis
Gómez Ríos, Yohima Valdés Núñez, Yanet Miguez
Barrios, Lisis Morales Viña, Ana María Cinza González.
Introducción del paquete documental para la producción
de Factor de Crecimiento Epidérmico humano
recombinante en el Bloque de Liofilizados.
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• Belkis Lérida Abreu Piñeiro, Yohanix López García,
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Oscar Cruz Gutiérrez, Gustavo Furrazola Gómez,
José García Suárez, Natacha Pérez Rodríguez, Osnel
García Roque, Daniel González Aguilar, Mónica M.
Navarro Mena, Denis Álvarez Betancourt, Yalina Ordaz
Contreras, Aleydis Gómez Ríos, Yanet Miguez, Barrios,
Yohima Valdés Núñez, Norelbys Albelo Rondón, Michael
Aldetere Ávalos, Rubén Enrique López Edghill. Proceso
de homologación de materias primas del Bloque de
liofilizados.
Caridad Suárez Martínez, Ilena García, Ivonne
Rodríguez Lima, Abel Caballero Pérez, Juan Miguel
Rivera, Francis Hernández, Manuel Pimentel, Yamile
Mendoza, Pedro Domínguez, Carlos Martínez Laporte,
Yeny de la Torre Maya, Rafael Fernández Almirola,
Miraidis Pérez Rizo, Iván Campa Legra. Aplicación de
la administración de riesgo a la calidad en las áreas de
producción de Planta 1.
David Diago, Gustavo Furrazola, Livan Maseda, Noel
Reyes, Arnaldo García, Gudelia Pérez, Galina Moya,
Mayda Candelario, Mayda Martínez, Karelia Cosme,
Iriac Bisquet, Lourdes Zumalacarregui, Juana Maria
Hernández, Josefina Astorga. Realización de los lotes
de garantía y confección del informe final. Adecuación
y entrega de la versión final de la documentación
al Instituto Pasteur de Irán (IPI) del Ingrediente
Farmacéutico Activo de la vacuna recombinante contra
el virus de la hepatitis B (IFAHB) en la planta PHB.
David Diago, Gustavo Furrazola, Livan Maseda, Noel
Reyes, Arnaldo García, Gudelia Pérez, Galina Moya,
Mayda Candelario, Mayda Martínez, Karelia Cosme,
Iriac Bisquet, Lourdes Zumalacarregui, Juana Maria
Hernández, Josefina Astorga. Realización de los lotes
de garantía y confección del informe final. Adecuación y
entrega de la versión final de la documentación al Instituto
Pasteur de Irán (IPI) del Ingrediente Farmacéutico
Activo de la Estreptoquinasa recombinante (IFASK) en
la planta PSK.
David Diago, Gustavo Furrazola, Mayté Pérez, Haydee
Gerónimo, Noel Reyes, Arnaldo García, Gudelia Pérez,
Galina Moya, Mayda Candelario, Mayda Martínez,
Karelia Cosme, Iriac Bisquet, Lourdes Zumalacarregui,
Juana Maria Hernández, Josefina Astorga. Realización
de los lotes de garantía y confección del informe
final. Adecuación y entrega de la versión final de la
documentación al Instituto Pasteur de Irán (IPI) del
Ingrediente Farmacéutico Activo de la Eritropoyetina
recombinante (IFAEPO) en la planta PEPO.
• David Diago, Isabel Apezteguía, Nubia González,
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Mariela Pérez, Ailen Sanchez, Odalys Ruiz, Lian Trimiño,
Marbelis Linares, Miladys Limonta, Eduardo Martínez,
Jorge Valdés Hernández, Gabriel Marquez y Dinorah
Torres Idavoy. Diseño y organización de un registro
maestro de producción para la etapa de desarrollo
tecnológico. Aplicación en las campañas productivas
2010-2011.
Denis Alvarez Betancourt. Revalidación de la producción
del IFA de P64Kr.
Denis Álvarez, Alejandro Beldarraín, Amado León,
Mónica Navarro. Introducción de la Proteína P64Kr
en Plantas Multiproducto y establecimiento de una
metodología cumpliendo con los requerimientos de
proceso y regulatorios.
Denis Alvarez, Monica Navarro Mena, Amado León
Cabrera, Oleg E. Alegret Savinova, Luciano Hernández
Marrero, Regla Margarita Hernandez Somoza.
Modificación del proceso de centrifugación/Precipitación
de la proteína P64Kr utilizando centrífugas tubulares..
Diamilé González Roche y Yanay Proenza Jiménez.
Obtención de una construcción genética negativa del
plasmidio que contiene el gen que codifica para la
proteína HBcAg, del proyecto Nasvac.
Diamilé González Roche, Yanay Proenza Jiménez,
Zeila Santana Vázquez, Adelina Pérez Chaviano.
Secuenciación del gen codificante para la proteína
HBcAg (antígeno vacunal del proyecto Nasvac. Análisis
de control del Banco de Células Primario extendido
071212 (BCPe 071212).
Diamilé González Roche, Yanay Proenza Jiménez,
Zeila Santana Vázquez, Adelina Pérez Chaviano.
Secuenciación del gen codificante para la proteína
HBcAg (antígeno vacunal del proyecto Nasvac. Análisis
de control del Banco de Células de Trabajo 071212
(BCT071212).
Eneida Roca, Jorge Sanchez, Lourdes Zumalacarregui,
Yanet Terrero, Gabriel Marquez, Neyda Hernandez,
Roxana Hernandez. Reajuste de los parámetros de
operación en la cromatografía de afinidad por iones
metálicos para la purificación de la proteína contra el
virus del dengue tipo 2.
Francis Hernández, Yunior Ramírez, Boris Menéndez,
Angel W. Quintana, Yoel A. Madruga, Nelvis Lamar,
Juana M. Hernández, Yenay Díaz, Ivonne Rodríguez,
Caridad Suárez, Nancy Pentón, Juan M Rivera.
Demostración del tiempo de vigencia de seis soluciones,
empleadas en la purificación del HBsAg, filtradas a
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través de cápsula y envasadas en frascos de vidrio de
20 L.
Francisco Castañeda Marquez, Emilio Narciandi Diaz,
Carlos Del Moral Pelier, Manuel Montane Enriquez,
Jorge Luis Vega Elias, Ricardo Ricardo Parellada,
Karelia Cosme Diaz, Yair Quiñones Maya, Luis Alvarez
Alvarez. Recapitalización de capacidades productivas,
tecnológicas y de mantenimiento del sistema productivo.
Gerardo Ramsés Hernández González, Jorge
Sotolongo Peña. Kosara Sánchez Álvarez. Utilización
de las técnicas de liomicroscopia y ATD/Z para la
caracterización de las propiedades térmicas de la
formulación liofilizada de la Estreptoquinasa humana
recombinante desarrollada por el CIGB.
Gerardo Ramsés Hernández González, Raymersy
Aldana Wilson, Sheila Padrón Morales, Nestor S.
Expósito Raya, Mabel Izquierdo López, Mariela Pérez
de la Iglesia, Yanet Terrero Socorro, Inalvis Herrera
Rivas, Yayri Caridad Prieto. Evaluación de la estabilidad
del antígeno en la vacuna terapéutica CIGB-247A.
Gustavo Furrazola Gómez, Eleine Mesa Pérez,
Dianelys Cabrera Pérez, Juana María Hernández
Rodríguez, Natacha Pérez Rodríguez, Leonardo Pacín
Olivares, Alejandro Hernández Paglieri, Yadira Mora
López y Oscar Cruz Gutiérrez. Validación del proceso
de filtración final esterilizante del IFA de IFN gamma
humano recombinante.
Gustavo Furrazola, David Diago, Liván Maseda,
Noel Reyes, Arnaldo García, Gudelia Pérez, Galina
Moya, Haydee Gerónimo, Mayda Candelario, Mayda
Martínez, Karelia Cosme, Iriac Bisquet, Lourdes
Zumalacarregui, Juana María Hernández, Josefina
Astorga. Realización de los lotes de garantía y
confección del informe final. Adecuación y entrega de
la versión final de la documentación al Instituto Pasteur
de Irán (IPI) del Ingrediente Farmacéutico Activo del
Interferón Alfa 2b humano recombinante (IFAIFN) en
la planta PIFN.
Iriac Bisquet Ramírez, Martha Aida Cabrisas Alfonso,
Manuel Montané Enríquez José Angel Acosta Buxado,
Armando Andrés Duarte. Comportamiento de los
portadores energéticos y sus tendencias en la Planta
de producción del CIGB durante el período 2008-2012.
Iriac Bisquet Ramírez, Martha Aida Cabrisas Alfonso,
Marielys Bello, Leoner del Arco, Manuel Montané
Enríquez, Armando Andrés Duarte. Comportamiento
de algunos indicadores del plan de Mantenimiento en
la planta de Producción del CIGB durante el año 2012.
• Isabel Apezteguía Rodríguez, David Diago Abreu, Nubia
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González de Armas, Tania González López, Milagros
Rodríguez Rodríguez, Dinorah Torres Idavoy, Eduardo
Martínez Díaz. Incorporación de las áreas de Diseño y
Desarrollo en el CIGB al Sistema de Gestión de Calidad
del CIGB.
Jorge Gallardo Alfonso, Elías Nelson Rodríguez, Maelys
Miyares, Sheyla Padrón, Bárbara María Quevedo.
Evaluación de la ruptura celular con molino de bolas en
el proceso de obtención del AgcHB.
Lázaro Estenoz Cosme, Yaquelín Santana Coto,
Aymara Quiala Mojena, Yosvani Abreu Prieto, Leonardo
Pacín Olivares, Ariel Rodríguez Rodríguez, Darién Rojo
Calvo, Jorge A. Presno Menéndez, Mario A. Cuadra
Valdes, Eleine Mesa Pérez, Dari Fasco Tornes, Dianelis
Cabrera Pérez, Jose A. Diaz Laurencio, Osnel Garcia
Roque, Aismara de la C. Morán Abreu, Yamilka Peña
Méndez, Lorena Perez Rodríguez, Leonardo Perez
Benitez. Informe final de la ejecución del cambio de
campaña de G-CSF a Interferón gamma en Planta 5.
Lissette López, Neyda Hernandez, Tatiana Gonzalez,
Bárbara Perez, Annette Pereira, Alina García, Ileana
Rosales, Julio C Sánchez. Validación de la técnica de
ELISA para la cuantificación del antígeno de superficie
del virus de la Hepatitis B recombinante (HBsAg-r)
con el anticuerpo monoclonal CB.Hep-1 en muestras
procedentes del proceso de producción que se analizan
en el Departamento de Control de Procesos.
Lissette López, Neyda Hernandez, Tatiana Gonzalez,
Bárbara Perez, Annette Pereira, Alina García, Ileana
Rosales, Julio C Sánchez. Utilización de las técnicas
de liomicroscopia y ATD/Z para la caracterización de
las propiedades térmicas de la formulación liofilizada de
la Estreptoquinasa humana recombinante desarrollada
por el CIGB. Control de Procesos.
Liurdis Caballero González, Rolando Páez Meireles,
Anazuria Martínez Nuñez, Yadir Cruz, Yanet Terrero,
Inalvis Herrera, Fidel Raúl Castro, Elian Cruz Peñalver.
Caracterización del proceso de obtención del GCSFpegilado a escala analítica.
Mayra Wood Duque, Neyda Hernández Caso, Margela
Montañez Valdés, Julio C. Sánchez García. Validación
de la cuantificación de proteínas totales, por el método
de Lowry, a muestras de la producción de la vacuna
sintética contra el Haemophilus influenzae.
Mayra Wood, Neyda Hernández, Margela Montañés,
Julio C. Sánchez, Josefa Ramos. Validación del método
de Ellman para la determinación de la concentración de
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Grupos Sulfidrilos, a muestras de la producción de la
vacuna sintética contra el Haemophilus influenzae tipo b.
Oleg Evio Alegret Savinova, Julio Cesar Yanes
Naranjo, Raudel Sosa Echagarruga, Yohanix López
García, Ronald de Oro Jorge, Daniel Gonzalez Díaz,
Rolando Martell Aedo, Yadira Mora López, Yamiris
Suarez Saavedra, Oscar Cruz Gutierrez, Denis Alvarez
Betancourt, René Meynardiez Rivero, Armando Álvarez
Alonso, Marcos Felipe Moreno Miravalles, Luís Mariano
Parreño Valdespino, Yaisel Nuñez Kairus, Orlando
Cabrera García. Validación de los procedimientos de
limpieza del equipamiento no dedicado en los cambio
de campaña de Planta 6, concluida la producción de
FCE-hr para uso tópico.
García, Mayte de la Caridad Perez Caballero, Yadira
Mora López, Yamiris Valido Artiles, Armando Álvarez
Alonso, Marcos Felipe Moreno Miravalles, Luís Mariano
Parreño Valdespino, Yaisel Nuñez Kairus y Oscar Cruz
Gutierrez. Validación de los procedimientos de limpieza
entre lote del equipamiento utilizado en la Etapa de
Ruptura celular correspondiente a la producción de SKr
en Planta 6.
García, Ronald de Oro Jorge, Mayte de la Caridad
Perez Caballero, Yadira Mora López, Yamiris Valido
Artiles, Oscar Cruz Gutierrez, René Meynardiez Rivero,
Armando Álvarez Alonso, Marcos Felipe Moreno
Miravalles, Luís Mariano Parreño Valdespino, Yaisel
Nuñez Kairus, Orlando Cabrera García y Denis Alvarez.
Validación de los procedimientos de limpieza entre lote
del equipamiento utilizado en la Etapa de Fermentación
correspondiente a la producción de P64Kr en Planta 6.
Oleg Evio Alegret Savinova, Julio Cesar Yanes Naranjo,
Yohanix López García, Oscar Cruz Gutierrez, René
Meynardiez Rivero, Armando Álvarez Alonso, Luís
Mariano Parreño Valdespino, Yaisel Nuñez Kairus,
Orlando Cabrera García, Nelvis Lamar Herrera, Cilia
María Mederos Robaina, Raudel Sosa Echagarruga,
Lisi Morales Viña, Yamiris Valido Artiles, Orlando
Cívico Dávalos, Iriac Bisquet Ramirez y José Garciga
Caballero. Informe final de la ejecución del cambio de
campaña de P64Kr a G-CSF en Planta 6.
Oscar Cruz Gutiérrez, Abel Caballero Pérez, Luciano
Francisco Hernández, Denis Alvarez Betancourt,
Ernesto Urrutia Váldez, Natacha Pérez Rodriguez,
•
•
•
•
Mónica Navarro Mena, Oleg Evio Alegret Savinova,
Lázaro Estenoz Cosme, Yohanix López García, Marbel
Ramos Alfonso, Mercedes Ortega Pérez, Kenia Vázquez
Montero, Juana María Hernández, Yenay Díaz León,
Lourdes Costa Anguiano, Marisel Quintana Esquivel,
Julio César Sánchez, Rafael del Toro Llanez, Jorge
Luis López Reconde, Manuel Montané Enriquez, Iriac
Bisquet Ramirez, Tania de la Cruz Curbelo. Validación
del proceso de producción para la obtención del
Ingrediente Farmacéutico Activo de IFN alfa 2b Humano
recombinante.
Rafael Fernández Almirola, Ivonne Rodríguez Lima,
Yeny de la Torre Moya, Yoel Madruga González,
Caridad Suarez Martinez, Carlos Martinez Laporte,
Miraidis Pérez Rizo. Desempeño de la higienización de
los sistemas tecnológicos de Planta 1. Producción de
HBsAg como IFA.
Rafael Fernández, Ivonne Rodríguez, Yeny de la Torre,
Yoel A. Madruga, Gregoria C. Suárez, Carlos B. Martínez,
Juan M. Rivera, Miraidis Pérez, Pedro Domínguez, Adrián
Núñez, Adriana Hernández, Agustín Blanco, Alexander
Giraud, Ángel W. Quintana, Abel Caballero, Aramis
Fernández, Ariam Valdés, Ariel Mesa, Arleidis Hinojosa,
Boris Menéndez, Carlos Peña, Cirenayka Belot, Danae
Fasco, Derman Trujillo, Elia Lias, Elvis Bilbao, Dagoberto
Sacereo, Fernando Ferrer, Giosvanys Vargas, Gretchen
Martínez, Imandra Ilizastegui, Julio Alberto Valdés,
Yunior Ramírez, Leandro Godínez, Liuven E. Guerrero,
Liván Ortega, Luis L. Pérez, Manuel Alberte, Manuel
Rodríguez, Miguel A. Pérez, Osvaldo Ávila, Pedro
Pérez, Vladimir Oceguera, Yamilé Mendoza, Yendry De
la Paz, Yohangel Guevara, Yordan I. Rojas, Yordanis
Pérez, Humberto Céspedes, Francis Hernández, José
L. Rodríguez, Nancy Pentón, Yordanis A. Gómez,
Edgar González. Desempeño de la higienización de los
sistemas tecnológicos de Planta 1. Producción de HBsAg
como IFA. Campaña 2011 – 2012.
Raymersy Aldana Wilson, Gerardo Ramsés Hernández
González, Nestor S. Expósito Raya, Sheila Padrón
Morales, Inalvis Herrera Rivas, Mariela Pérez de la
Iglesia, Yayri Caridad Prieto. Evaluación y selección del
adyuvante tipo alúmina a utilizar en la preparación de la
vacuna terapéutica CIGB-247A.
Rebeca Bouyón Albarrán, Raudel Sosa Echagarruga,
Julio César Sánchez García, Hismelys Díaz Ramírez,
Amado León González, Iberty Ramírez Arcia. Estudios de
recobrado para la validación de la limpieza e higienización
de equipos en la producción del IFA FCE hr.
114
YEARBOOK 2013
PUBLICATIONS
• Tania de la Cruz Curbelo, Martha A. Cabrisas Alfonso, Iriac Bisquet
•
•
•
•
•
•
•
•
•
•
Ramírez, Manuel Montané, Armando Duarte, Marielys Bello Gil.
Evaluación de la actividad de Mantenimiento Tecnológico de la
Planta de Producción como parte del proceso de Equipamiento e
Infraestructura del Sistema de Gestión de la Calidad en el año 2012.
Yinet Cartaya Rodríguez; Jorge Sanchez Romeu; Lourdes
Zumalacárregui de Cárdenas; Yamile Vega Hurtado; Rolando Perdomo
Morales; Gabriel Márquez Perera; Lazara Muñoz Hernández; Lidia
Gómez Terry. Estudio de la influencia de lavados con detergente
Tritón X 114 en la cromatografía de IMAC como paso de remoción de
endotoxinas en el proceso de purificación del CIGB-166.
Yoel Alberto Madruga González, Ivonne Rodríguez Lima, Rafael
Fernández Almirola, Arleidis Hinojosa Romero, Alexander Giraud
Lugo, Manuel Alberte Magriñá, Yordanys Gómez Rodríguez, Pedro
Domínguez Aldás, Manuel Rodríguez Pimentel, Yamilé Mendoza
Gutiérrez, Francis Hernández Coro, Nancy Pentón Piña y Juan
Miguel Rivera Martín. Desempeño de las matrices cromatográficas
en la producción de HBsAg como Ingrediente Farmacéutico Activo
(IFA), durante la campaña 2012.
Yohanix López García, Oleg Evio Alegret Savinova, René Meynardiez
Rivero, Julio Cesar Yanes Naranjo, Cilia María Mederos Robaina,
Mayte de la Caridad Perez Caballero, Yadira Mora López, Oscar Cruz
Gutiérrez y Yamiris Valido Artiles. Validación del tiempo de vigencia de
las soluciones utilizadas en la producción de IFN gamma en Planta 6.
Yoymí Claro, Diamilé González, Jorge Valdés, Michel Díaz, Ernesto
Mantilla, Alain Salazar, Yanay Proenza. Relación entre el nivel de
expresión y el número de copias del gen del Antígeno de Superficie
del Virus de la Hepatitis B presentes en la cepa C20 de Pichia pastoris.
Yoymí Claro, Diamilé González, Mariela Pérez, Jorge Valdés, Michel
Díaz, Yanay Proenza, Ernesto Mantilla, Alain Salazar. Estudio de la
estabilidad del gen codificante para el Antígeno de Superficie del
Virus de la Hepatitis B en el Banco de Células Primario (BCP090542).
Diseño, construcción y escalado de un mezclador de laboratorio
para la preparación de emulsiones.
Obtención de un modelo matemático para el diseño de formulaciones
oleosas estables con el uso de Montanide 888 VG.
Expediente de Registro Sanitario para la Renovación de Heberprot-P®
en Cuba.
Expediente para Registro Sanitario de Heberprot-P® en Venezuela.
Expediente de Registro Sanitario para la Renovación de Heberprot-P®
en Vietnam.
PATENTS
• Centro de Ingeniería Genética y Biotecnología de Sancti
Spíritus, propietario. Método de Obtención de 1-kestosa. PCT/
CU2013/000005.
SUMMARY
115
YEARBOOK
2013
CENTER FOR GENETIC ENGINEERING
AND BIOTECHNOLOGY

yearbook 2013

Transcription

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