Progesterone/Denise Mullery - UC Davis Department of Chemistry

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Progesterone/Denise Mullery - UC Davis Department of Chemistry
Chemistry 150, Fall 2008
“The Pill”: The Occurrence,
Biological Activity, Biosynthesis,
and Synthesis of Progesterone
(www.diplom-biologe.de/images/meiner.jpg)
Denise Mullery, Fall 2008
Introduction
Progesterone originates from the yam family, Dioscorea.
Dioscorea produces large amounts of a steroid called
diosgenin, which can be converted into progesterone.
Progesterone plays a major role in pharmaceuticals by being a
significant component of birth control pills to prevent women
from becoming pregnant. There is evidence of biosynthesis of
converting diosgenin into progesterone. As for synthesis of
progesterone, there are many reactions that have been tested
and produced variable yields, but only a couple of them will
be discussed later on in this final paper. Along with the
synthesis, the occurrence, biological activity, and biosynthesis
of progesterone will also be talked about in this final paper.
O
H
H
H
O
Progesterone
Figure 1.
Dioscorea.
Progesterone,
the
steroidal
triterpenoid
of
I. Occurrence
Progesterone is mostly, but indirectly found in Dioscorea
mexicana. Dioscorea mexicana is a plant that is part of the
yam family and resides in Mexico.1 It contains a steroid
called diosgenin that is taken from the plant and is converted
into the valued product, progesterone. 2 On the other hand,
diosgenin and progesterone are not only found in Dioscorea
mexicana, but also in other Dioscorea species.
Figure 1. Dioscorea mexicana. Source
Figure 2. Birth control pill. Source
(http://www.canadiansforchoice.ca/images/contraception_img
03.gif)
Another plant that has been discovered to indirectly contain
progesterone is Dioscorea pseudojaponica that resides in
Taiwan. Research has showed that the Taiwanese yam
contained saponins, steroids, which are converted to
diosgenin. From diosgenin, it can produce progesterone. 3
Another plant that is found to contain these steroids that
convert to progesterone is a wild yam called Dioscorea
villosa. One study showed that the Dioscorea villosa contains
3.5% of diosgenin.4 There are many other Dioscorea species
of the yam family that contribute to the production of
progesterone, but there is one new species found. Dioscorea
polygonoides has been researched and found 2.64% of
diosgenin that was recognized by testing it with gas
chromatography-mass spectrometry. 5 Many of the Dioscorea
species that originate from the yam family grow in countries
that have tropical and subtropical climates. 6
II. Biological Activity
Progesterone is a steroid hormone that plays a major part in
female pregnancy. 7 Varying responses to transcription activity monitors it. Transcription activity is determined by two
progesterone receptors, PR-A and PR-B. Both originate from
the same gene and are somewhat the same, but for the most
part vary on reactions to progesterone. 8 In one study, “PR-A
protein shows normal mammary gland response to
progesterone but severe uterine hyperplasia and ovarian
abnormalities…PR-B protein does not affect biological
responses of the ovary or uterus to progesterone but results in
reduced pregnancy-associated mammary gland morphogenesis.”7 Several studies have shown that antiprogestins prevent progesterone receptors from proceeding the pregnancy
process.8
Both progesterone receptor proteins, PR-A and PR-B, have
“a discrete hormone-binding domain close to a blocked amino
acid terminal region of the peptide chain.”9 Since both proteins came from the same gene, they have similar codes for
binding to hormones, but still vary in other parts of the gene.9
One of the differences between hPR-A and hPR-B is the
carboxyl group at the end of hPR. Human progesterone
Chemistry 150, Fall 2008
receptor-B (hPR-B) starts the transcription of progesterone
genes, while hPR-A blocks the transcription of progesterone
receptors. An example of this was tested in a study where
“hPR-A mediated repression of human estrogen receptor
(hER) transcriptional activity is not dependent on hER
expression level but depends largely on …expression level of
hPR-A.”10 hPR-A is able to stop transcription of hER because
there is no competition to prevent hPR-A from performing its
function.
hPR has agonists and antagonists that are attached to each
other by ligand-receptors. A study proved that “a third class
of hPR ligands that induce a receptor conformation which is
distinct from that induced by agonists or antagonists.”11
Genes that react to the progesterone receptors are partial
agonists. Overall, this shows that different ligands networking in different areas of different binding locations of
PR’s will vary in multiple biological outcomes. 11 One example of this was a study on rats being tested with
gonadotropin (hCG). In the study, “hCG stimulated the production of progesterone and this effect was blocked by
tyrosine… The essential functional groups necessary for progesterone production antagonism were they tyrosine amino
and phenolhydroxyl groups, whereas the carboxyl group was
seemingly essential for the antagonistic effect.” 12 Another
example of having varied processes, but a common result is
the study of the level of affinity for progesterone receptors.
In this study, 19-Nor-progesterone (19NP) is a “potent
progestagen, which possesses a high affinity for the progesterone receptor. In contrast, 17α-hydroxylated-progesterone
(170HP) had no hormonal activity and does not compete for
the progesterone receptor.”13 One way to achieve a high
affinity for 170HP is “the attachment of a 17α-hydroxyl group
on 19NP…addition of a Me group combined with the
formation of a double-bond at C6 results in nomegestrol
(NOM)…and the protection of the free 17α-hydroxyl group of
NOM by an acetate led to a potent progestin with high
affinity.” 13 One way to measure the affinity of progesterone
receptors attaching to steroids is by using the Minimal
Topological Difference (MTD) method. This method measures the hydrophobic level of progesterone to see if it has
high chances of bonding to the steroid. 14
These attachments between PR and steroid are related to
the human body by the uterine endometrium and myometrium
uniting with the progesterone receptors. This was proven in a
test where “the gonane progestogen dl-norgestrel bound
strongly to the uterine receptors, but only the d-isomer was
active. All three pregnane progestogens, i.e. chlormadinone
acetate, medroxyprogesterone acetate, and megestrol acetate
also showed significant binding to receptors, but for estrane
progestogens, only norethisterone had a high binding
capacity.” 15
Overall, in order to see if progesterone was effective in
birth control pills, a study was commenced. In one study, “50
[women] took combined low-dose estrogen and progesterone
pills for a year or more and the other 50 used different
methods of birth control.” 16 In the end, there were no
negative effects of taking the progesterone pills for birth
control. As a word of caution, if the progesterone receptors
are disrupted during the first 28 days of pregnancy, there will
be negative consequences towards the ability to reproduce. 17
III. Biosynthesis
Progesterone can be biosynthetically produced from
steroidal diosgenin, which originates from triterpenoids.
Diosgenin undergoes acetylation to make 2. It then oxides to
produce 3 and performs ester hydrolysis to form
dehydropregnenolone acetate 4. Afterwards, hydrogenation
turns 4 into pregnenolone acetate 5. Then a hydroxyl group is
added to replace the -OAc group to form pregnenolone 6.
Lastly, Oppenauer oxidation and tautomerism occurs to
produce progesterone.18 This reaction is very similar to
converting cholesterol converting into progesterone based on
the fact that both reactions rely on oxidation to form
progesterone. 19 A study has proven that pregenolone and
progesterone can also be biosynthesized by yeast. 20
O
H
O
H
H
H
H
HO
O
H
Ac2O
heat
H
2
O
H
O
H
H
AcO
4, dehydropregnenolone
H2/Pd
acetate
3
O
H
H
AcO
O
H
OHH
5, pregnenolone
acetate
H
HO
AcO
O
O
H
HOAc
heat
H
H
AcO
CrO3
H
H
AcO
1, diosgenin
AcO
H
Al(OPr)3
Me2CO
H
6, pregnenolone
O
H
H
O
Figure 3. Biosynthesis of progesterone18
H
progesterone
IV. Synthesis
The synthesis of progesterone begins with 7 reacting with
PhLi in THF to produce 8. Then it mixes with the reagent of
step 1 while the temperature increases from -70°C to -30°C
and follows it up with reacting again with PhLi along with a
surplus amount of MeOH at -30°C to make 9. Afterwards, 0.1
N HCl at an increased temperature of 40°C is added into the
reaction that leads to 10. EtOH and 2% of NaOH then close
the ring to form 11 yielding 40%. The next step is the
reaction undergoes methylation and hydroxylation to develop
12 at a high yield of 99%. The reactant next transforms into
chair conformation 13 after reacting in TFA at 0°C. In the
following step, it undergoes cation-π cyclization to produce
14. The next step produces 15 after yielding 72% by breaking
off the cation. After that, the ring is opened and oxidized at -
Chemistry 150, Fall 2008
70°C and then increases from -15°C to 23°C to produce 16.
Finally, it reacts with water and 5% of KOH with a ratio of
5:2, at 23°C, to produce progesterone at a decent yield of
51%.21
lower production amount of 78%.22
Me
O
PPh3+I-
Me
O
O
O
O
PPh3 1.
Me
O
O
Me
0.1 N HCl in H2O/
MeOH (3:10), 40°C
10
O
O
Me
Me
O
Me
Me
Me
11
O
MeLi,
Et2O
(99%)
Me
O
Me
O
H
H
Me
OH
TFA, 0°C
12
Me
Me
K2CO3,
H2O/MeOH
(72% from 12)
H
H
H
O
Me
Me
H
O3, MeOH/CH2Cl2,
70°C; then Zn, AcOH
H2O, -15°C to 23°C
(88%)
O
15
Progesterone had and still does play a major role in the
pharmaceutical world of birth control. Millions of women use
it to prevent pregnancy. Preventing pregnancy is not the only
benefit of using birth control pills. It also reduces acne,
protects against certain types of cancer related to the
reproductive system, adjusts and keeps the menstrual cycle in
order, etc. On the other hand, there are negative side effects
such as weight gain, headaches, etc. In some countries,
women can now buy the “morning-after pill”, or “Plan B”,
over-the-counter. This has only been passed in India, Canada,
Sweden, Norway, and the Netherlands. There has been some
recent news of making a birth control pill for men, but it is
currently undergoing clinical trials. The one factor is it
cannot be taken as a “pill”, but by skin cream or injection.
Overall, progesterone as an important component of the birth
control pill is commonly used around the world and will
continue to grow as it becomes more available to the public.
13
Me
O
Me
Me
Me
cation-!
cyclization
14
Me
O
Cl(CH2)2Cl,
O
O
H
Me
Me
O
9
Me
2, progesterone
90% (CHCl3)
78% (CH3CN)
Conclusion
Me
Me
O
H
O
Figure 5. Semi-Synthesis of Progesterone22
O
-70°C to -30°C
8
Me
Me
Me
H
H
H
1, desoxycorticosterone
2. PhLi (1 equiv.); then
excess MeOH, -30°C
O
7
EtOH/2% NaOH
in H2O, reflux
(40% from 1.)
Me
Me2SiI, solvent
O
Me
THF, PhLi
(1 equiv.
in Et2O)
OH
O
Me
O
O
Me
H
H
Me
O
Me
Me
________________
H
H 2O
5% KOH (5:2),
23°C
(51%)
H
H
O
Me
References
Me
H
16
1
Applezweig, N. Steroids. Chem. Week 1969, 104, 57-72.
2
Noguchi, E.; Fujiwara, Y.; Matsushita, S.; Ikeda, T.; Ono,
M.; Nohara, T. Metabolism of Tomato Steroidal Glycosides
in Humans. Chem. Pharm. Bull. 2006, 54, 1312-1314.
3
Yang, D. J.; Lu, T. J.; Hwang, L. S. Isolation and
Identification of Steroidal Saponins in Taiwanese Yam
Cultivar (Dioscorea Pseudojaponica Yamamoto). J. Agric.
Food. Chem. 2003, 51, 6438-6444.
4
Hooker, E. Final Report of the Amended Safety Assessment
of Dioscorea Villosa (Wild Yam) Root Extract. Int. J.
Toxicol. 2004, 23, 49-54.
5
Nino, J.; Jimenez, D. A.; Mosquera, O. M.; Correa, Y. M.
Diosgenin Quantification by Hplc in a Dioscorea
Polygonoides Tuber Collection from Columbian Flora. J.
Braz. Chem. Soc. 2007, 18, 1073-1076.
6
Myoda, T.; Nagai, T.; Nagashima, T. Properties of Starches
in Yam (Dioscorea Spp.) Tuber. Curr. Top. Food. Sci.
Technol. 2005, 105-114.
O
Me
Me
Me
H
H
H
H
O
progesterone
Figure 4. Synthesis of progesterone21
The next semi-synthesis of progesterone is much simpler
than the previous one because it will only focus on the
regioselectivity. In this semi-synthesis, there are two ways to
produce progesterone, but each way ends with a different
yield. For the first semi-synthesis, desoxycorticosterone
reacts with Me3 SiI in CHCl3 to lose a hydroxyl group and
produce progesterone. Using CHCl3 as a solvent, it gives the
product at lower yield, but it produces progesterone at a
greater amount of 90%. As for the second semi-synthesis,
descoxycorticosterone reacts with Me3 SiI in MeCN to make
progesterone as a result of losing a hydroxyl group. MeCN as
a solvent produces progesterone at a higher yield, but at a
Chemistry 150, Fall 2008
7
8
Mulac-Jericevic, B.; Conneely Orla, M. Reproductive Tissue
Selective Actions of Progesterone Receptors. Reproduction
2004, 128, 139-146.
20
Duport, C.; Spagnoli, R.; Degryse, E.; Pompon, D. SelfSufficient Biosynthesis of Pregnenolone and Progesterone in
Engineered Yeast. Nat. Biotechnol. 1998, 16, 186-189.
Gadkar-Sable, S.; Shah, C.; Rosario, G.; Sachdeva, G.; Puri,
C. Progesterone Receptors: Various Forms and Functions in
Reproductive Tissues. Front Biosci. 2005, 10, 2118-2130.
21
Johnson, W. S. Synthesis of Progesterone J. Am. Chem. Soc.
1971, 4332.
22
Numazawa, M.; Nagaoka, M.; Kunitama, Y. Regiospecific
Deoxygenation of the Dihydroxyacetone Moiety at C-17 of
Corticoid Steroids with Iodotrimethylsilane. Chem. Pharm.
Bull. 1986, 34, 3722-3726.
9
Birnbaumer, M.; Weigel, N. L.; Grody, W. W.; Minghetti, P.
P.; Schrader, W. T.; O'Malley, B. W. Progesterone Receptor:
Structure and Function. Progesterone Progestins, [Int.
Symp.] 1983, 19-32.
10
Wen, D. X.; Xu, Y.-F.; Mais, D. E.; Goldman, M. E.;
McDonnell, D. P. The a and B Isoforms of the Human
Progesterone Receptor Operate through Distinct Signaling
Pathways within Target Cells. Mol. Cell. Biol. 1994, 14,
8356-8364.
11
Wagner, B. L.; Pollio, G.; Leonhardt, S.; Wani, M. C.; Lee,
D. Y. W.; Imhof, M. O.; Edwards, D. P.; Cook, C. E.;
McDonnell, D. P. 16ϱ-Substituted Analogs of the
Antiprogestin Ru486 Induce a Unique Conformation in the
Human Progesterone Receptor Resulting in Mixed Agonist
Activity. Proc. Natl. Acad. Sci. U. S. A. 1996, 93, 87398744.
12
Wang, X.; Zhang, Q.; Gao, E.; Cheng, Z. Effects of Tyrosine
and Its Analogs on Hcg-Induced Progesterone Production by
Rat Corpus Luteum in Vitro. Chin. Med. J. (Beijing, Engl.
Ed.) 1987, 100, 859-864.
13
Botella, J.; Duc, I.; Delansorne, R.; Paris, J.; Lahlou, B.
Structure-Activity and Structure-Affinity Relationships of
19-nor-Progesterone Derivatives in Rat Uterus. J.
Endocrinol. Invest. 1990, 13, 905-910.
14
Bohl, M.; Simon, Z.; Vlad, A.; Kaufmann, G.; Ponsold, K.
Mtd Calculations on Quantitative Structure-Activity
Relationships of Steroids Binding to the Progesterone
Receptor. Z. Naturforsch., C: Biosci. 1987, 42, 935-940.
15
Briggs, M. H. Contraceptive Steroid Binding to the Human
Uterine Progesterone-Receptor. Curr. Med. Res. Opin. 1975,
3, 95-98.
16
Ishak, R.; Mohamed, A. B.; Hassan, K. Platelet Function
Studies in Women on Oral Contraceptive Pills. Malays J
Reprod Health 1990, 8, 1-4.
17
Ordian, N. E.; Akulova, V. K.; Mironova, V. I.; Pivina, S.
G.; Rakitskaia, V. V. Influence of Progesterone Receptors
Deficit in Early Stage of Development on Formation of the
Reproduction Functions of Female Rats. Ross Fiziol Zh Im I
M Sechenova 2008, 94, 465-473.
18
Dewick, P. Medicinal Natural Products: A Biosynthetic
Approach. 2nd ed.; Wiley&Sons: West Sussex, England,
2001; p 266.
19
Tialowska, B.; Klimek, J.; Zelewski, L. Progesterone
Biosynthesis Supported by Fatty Acid Oxidation in the
Mitochondrial Fraction of Human Term Placenta. Acta
Biochim. Pol. 1983, 30, 11-21.

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