As aplicações da espectrometria de massa no desenvolvimento de

Transcription

As aplicações da espectrometria de massa
no desenvolvimento de fármacos e
medicamentos
(Mass Spectrometry in the Development of
Drugs and Medicines)
Mike S. Lee !
Milestone Development Services!
!
IV Symposium!
Novas fronteiras farmacêuticas nas ciências, tecnologia,
regulamentação e sistema da qualidade!
Brasília, 22 e 23 de junho de 2015!
[email protected]!
Reference Materials – Mass
Spectrometry Applications in Drug
Discovery & Development
Integrated Strategies for
Drug Discovery Using
Mass Spectrometry
Mike S. Lee
ISBN: 978-0-471-46127-2
June 2005
John Wiley & Sons,
New York, NY
LC/MS Applications
In Drug Development
Mike S. Lee
ISBN: 978-0-471-40520-7
March 2002
John Wiley & Sons,
New York, NY
2
Reference Materials – Applications in
Drug Metabolism and Impurity and
Degradant Identification
Mass Spectrometry in
Drug Metabolism and
Disposition
Mike S. Lee and
Mingshe Zhu
ISBN: 978-0-470-40196-5
May 2011
John Wiley & Sons,
New York, NY
Characterization of
Impurities and
Degradants Using
Mass Spectrometry
Ben Pramanik, Mike S.
Lee and Guodong Chen
ISBN: 978-0-470-38618-7
May 2011
John Wiley & Sons,
New York, NY
3
Global Pharmaceutical Industry !
Perspectives!
§  Research & Development Cost!
-  No guarantee of success!
§  Market and Geographical Shift!
-  Aging population!
-  Prevention and diagnosis!
-  Asia!
-  Personalized medicine!
§  Diagnostics!
-  Accurate and efficient!
§  Biotechnology!
-  Genetic differences!
-  Precise treatment!
!
• On the Brink of a Healthcare Revolution!
!!
F.B. Humer, Hoffman-La Roche, 2005
Milestone Development Services
Worldview 1540!
Mass Spectrometry and
Chromatography for Healthcare!
§  Early detection!
-  Mass Spectrometer Transformed into a Clinical Analyzer!
-  A Molecular Microscope!!
§  Response-guided medication!
-  Chromatography-Mass Spectrometry Platforms Evolve into a
Clinical Analyzer!
-  A Molecular Plate Reader!!
!
!
• Mass Spectrometry and Chromatography Uniquely !
Positioned to Enable Personalized Healthcare!
- Integration, Miniaturization and Visualization are Critical for !
Success!!
!
Milestone Development Services
Analytical Focus on Industrial
Applications!
§ 
Qualitative!
§ 
§ 
§ 
Quantitative !
§ 
§ 
§ 
Sensitive, highly specific assays!
Trace-mixture analysis!
Mechanistic approaches!
§ 
§ 
§ 
Rapid structure confirmation!
Structure identification!
Immune response!
Reactive metabolites!
Industry Preferences for Quick Information !
and Fast Decision Making !
• Lee, LC/MS Applications in Drug Development 2002
!
7
Structure Analysis Matrix:
Pharmaceutical Analysis Preferences!
Pure
NonTrace
X-ray
NMR
Trace
MS
UV
IR
Mixture
LC/NMR
HPLC/UV
LC/MS
§ 
Trace-Mixture Sample Type is a Primary Driver for !
Accelerated Research and Development!
!
8
The LC/MS Flow Path!
Scale-Up &
Extraction
Fractionation
Autosampler
HPLC
Collection
UV
Spectroscopic Analysis
Tandem
Mass Spectrometer
§  The
Integration of Orthogonal Separation and!
Amplification Steps !
!
9
Analytical Figures of Merit –!
Qualitative and Quantitative Analysis!
Sensitivity
Sensitivity
Accuracy
Control
Sensitivity
Precision
Speed
§  Increase
Analytical Performance through Precise and !
Automated Control!
!
10
Mass Spectrometry Approaches for
Identification (LC/MS)!
§ 
Based on the fact that impurities, degradants, and metabolites
can retain substructures of the original molecule!
§ 
§ 
§ 
§ 
Full scan mass spectrometry allows for confirmation of
molecular weight!
§ 
§ 
Ionization (acid-base chemistry) provides facile confirmation!
Tandem mass spectrometry (MS/MS) provides rapid
identification of substructures!
§ 
!
Yost, Perchalski et al. Talanta 1984, 10, 929-935.!
Lee et al., Annu. Rep. Med. Chem. 1986, 21, 313-321.!
Lee and Yost, Biomed. Env. Mass Spectrom. 1988, 15, 193-204.!
Use the standard (i.e., parent drug) as a template!
§ 
Methodology provides a rapid and systematic approach without
the use of standards!
§ 
New Frontier (1980s)! Mass Spectrometry!
Destined to Industrialize Identification!
11
Limited Data Can Reveal Useful
Information - Particularly if it is Familiar!!
§  Mark
Cole, Pfizer (CPSA 2005)!
188
12
Summary of Identification Strategy!
§ 
Build the picture puzzle - stepwise!!
§ 
§ 
§ 
Think chemistry!
§ 
§ 
§ 
§ 
No need to identify all fragment ions to propose structure!
Address rearrangements later!
Total unknowns!
§ 
§ 
§ 
!
Exploit the fact that ionization is the initiator of fragmentation!
Acid-base chemistry!
Connect the pieces!
§ 
§ 
Molecular weight corresponds to the outside of the puzzle!
Direct comparison of homologous “pictures”!
Stop with the confident assessment of molecular weight!
High resolution/accurate mass (molecular formula) may help!
Isolation and scale up for NMR!
Lee, LC/MS Applications in Drug Development 2002
13
!
!
14
Metabolism Guided Structural
Modification!
§ 
§ 
§ 
Metabolite identification provides insights into
mechanisms of physiological degradation!
Early identification provides knowledge for
candidates!
Medicinal chemists can modify sites of metabolic
reactions to reduce deactivation and elimination!
15
Ionization/Fragmentation Template for
Buspirone!
1
2!
3!
O
N
N
N
N
N
6!
§  When
O
5!
4!
in doubt - every heteroatom except Carbon is basic!!
16
Tabulate Fragments and Formulas for
Buspirone!
Substructure!
1
!
!
2
!
!
3
!
!
4
!
!
5
!
!
6
!
Formula !
C17H30N3O2
C13H20NO2
C9H14NO2!
C12H19N4 !
C8H13N4 !
C 4H 3N 2 !
Mass
308
222
168
219
165
79
Observed (Y/N)!
!
N!
!
Y!
!
Y!
!
N!
!
N!
!
N!
17
Substructure - MS/MS Product Ion
Spectrum of Buspirone!
[M+H]+!
386!
!
122!
150!
265!
148!
222!
152! 180!
168!
109!
95!
198!
291!
343!
18
Create a Substructure Template for
Buspirone!
222+!
168+!
O
N
N
N
N
N
O
19
Identify Modified Substructure(s) Compound A !
238+
184+
O
N
N
N
N
N
O
+16 Da
20
Identify Modified Substructure(s) Compound B !
222+
168+
O
N
N
N
N
N
O
-78 Da
21
Structure Database!
§ 
§ 
§ 
§ 
Compilation of structural and analytical information!
Effective technology transfer!
Reference for rapid identification of analytes present
in future samples!
Overview of structural trends!
22
Structure Database - Buspirone!
!
!
Structure!
!
!
!RRT !
!MW!
!
!0.54 !
!401
!
!
!0.74 !
!307!
!
!
!
!
!1.0
!!
!385!
O
!
N
!
!
N
N
N
!
!
!
!
N
O
!
!!
!
+ OH
O
!
!
HN
N
!
!
!
N
O
O
!
!
N
!
!
N
N
N
!
!
N
O
!
23
Impurities in Drug Substance!
§ 
Drug substance!
§ 
§ 
§ 
An active ingredient that is intended to furnish pharmacological
activity or other direct effect in the diagnosis, cure, mitigation,
treatment, or prevention of disease or to affect the structure or any
function of the human body!
Does not include intermediates used in the synthesis!
Impurity!
§ 
Any component of the drug product that is not the chemical entity
defined as the drug substance or an excipient in the drug product!
24
Impurities in Drug Substance Thresholds!
Maximum
Daily Dose
!
≤ 2g/day
!
> 2g/day
§ 
!0.05%
!
!
!
!0.10% or 1.0 mg per day
!intake (whichever is lower)
0.15% or 1.0 mg per day!
intake (whichever is lower)!
!
!0.03%
!
!0.05%
0.05%!
!
Qualification !
Threshold!
A limit above (>) which an impurity should be identified!
The process of acquiring and evaluating data that establishes the biological safety of
an individual impurity or a given impurity profile at the level(s) specified!
Qualification threshold!
§ 
§ 
!
!
!
!
!
Qualification!
§ 
§ 
!Identification
!Threshold
Identification threshold!
§ 
§ 
!Reporting
!Threshold
A limit above (>) which an impurity should be qualified!
Impurity profile!
§ 
A description of the identified and unidentified impurities in drug substance!
Guidance for Industry:
Q3A Impurities in New Drug Substances
www.fda.gov
25
Identification of Impurities in Drug
Substance!
§ 
§ 
§ 
§ 
The impurity profile of a drug substance is one of the
most important quality characteristics!
Impurity profiles are almost always multivariate in
nature!
Impurity profiles are often complicated by the
relationships among impurity levels!
A summary of actual and potential impurities most
likely to arise during the synthesis, purification, and
storage of the new drug substance is required!
26
!
!
27
Paclitaxel!
O
!
!
O
O
OH
O
H
N
O
O
O
OH
H
HO
O
O
O
O
28
HPLC Chromatogram - Paclitaxel Bulk
Drug!
!
!
29
Paclitaxel!
O
O
O
OH
O
H
122
N
O
O
H
O
HO
OH
O
O
O
O
286
569
509
-CH3COOH
30
MS/MS Product Ion Spectrum of
Paclitaxel!
[M+H]+!
854!
!
O
O
O
OH
O
H
122
N
O
O
H
O
HO
OH
O
O
O
O
569
286
509
-CH3COOH
[M+NH4]+!
871!
286!
!
509!
268!
569!
551!
327!
776!
31
Base-Induced LC/UV Chromatogram!
32
Base-Induced Degradation Pathway!
33
Acid-Induced LC/UV Chromatogram!
34
Hydrogen Peroxide Incubation - LC/UV
Chromatogram!
35
1000 FC White Light (92 Days) -!
LC/UV Chromatogram!
36
Paclitaxel Structure Database!
Kerns et al., J. Nat. Prod. 1994, 57, 1391-1403
37
Database Applications!
§  Rapid dereplication of paclitaxel analogs in samples!
§  Retention time and MW often sufficient for rapid identification!
§  Structural insights from database compounds aid interpretation of new
analogs!
§  Process insights!
§  Structure provides chemical knowledge as basis for process
optimization!
§  Mechanism of impurity formation suggests changes in process!
§  Reduction of toxic impurities!
38
Degradant in Drug Product!
§ 
Drug product!
§ 
§ 
§ 
§ 
§ 
A finished dosage form that contains a drug substance,
generally, but not necessarily, in association with one or more
other ingredients (21 CFR 314.3(b))!
Tablet!
Capsule!
Solution!
Degradant!
§ 
A molecule resulting from a chemical change in the drug
molecule brought about over time and/or by the action of light,
temperature, pH, water, or by reaction with an excipient and/
or the immediate container/closure system
39
!
J Pharm Biomed Anal. 1996 Sep;14(12):1743-52
Predictive Strategy for the Rapid Structure Elucidation of Drug Degradants
Rourick RA, Volk KJ, Klohr SE, Spears T, Kerns EH, Lee MS
40
Sotalol!
41
Substructure Template of Sotalol!
43!
O
H3C
S
N
H
N
H
O
255!
OH
79!
42
Structure Database for Sotalol!
Rourick et al., J. Pharm. Biomed. 1996, 14, 1743-1752
43
!
44
Structure Database of Butorphanol
Degradants!
Volk et al., J. Pharm. and Biomed. Anal. 1996, 14, 1663-1674
45
Butorphanol Impurity Library!
Volk et al., J. Pharm. and Biomed. Anal. 1996, 14, 1663-1674
46
LC/MS Method Development Strategies!
§ 
§ 
§ 
§ 
§ 
§ 
§ 
§ 
§ 
§ 
Standard Methods!
Template Structure Identification!
Databases!
Screening!
Integration!
Miniaturization!
Parallel Processing!
Visualization!
Automation!
Powerful Tools to Devise, Construct and Refine an !
Analytical Method!
!
47
Clinical & Pharmaceutical Solutions through Analysis
August 3 – 5, 2015
São Paulo, Brazil
CPSA Vision CPSA -‐ Clinical and Pharmaceu5cal Solu5ons through Analysis represents a broad vision to promote new technologies and solu5ons across disciplines and forge global paths for educa5on and training. World-‐Class Programs Unique focus on industrial issues and needs featuring interna5onally recognized scien5sts and premier research. Thinking Big! Provide an ideal environment to benchmark a strategy or capture an epiphany. Big ideas, crea5vity and involvement are openly encouraged. Imagina5on needed! Diversity and Study Abroad Promote interdisciplinary educa5on, training and career development on a global level with unique focus on young scien5sts and students. Technological Rigor Support the highest level of technology evalua5on to assist professionals with the iden5ﬁca5on of prac5cal considera5ons to perform an analysis that meet speciﬁc regula5ons such as FDA or CLIA with the highest level of performance (accuracy, sensi5vity, throughput, cost). InteracAon and RelaAonships Facilitate the growing interest in industrial research with special events dedicated to promo5ng a collegial atmosphere and fostering friendship. CPSA is User Friendly! www.cpsa-brasil.com
Where Technology and Solutions Meet
August 3 – 5, 2015
São Paulo, Brazil
Global Community Langhorne, PA USA Shanghai, China São Paulo, Brazil www.cpsa-brasil.com
August 3 – 5, 2015
São Paulo, Brazil
2nd Annual Brazilian Symposium
On the Way to Innova5on: Pharmaceu5cal/Analy5cal Technology, Regula5on and Knowledge Management August, 3 to 5, 2015 – São Paulo/SP 2015 Program Chair Rafael Barrientos, Ph.D. Eurofarma Laboratórios S.A. www.cpsa-brasil.com

As aplicações da espectrometria de massa no desenvolvimento de

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