Weight loss abstract
A method for controlling weight gain or promoting weight loss which
includes the step of treating a subject with an effective weight
gain controlling or weight loss promoting amount of a substituted
.DELTA.5-Androstene which is biologically effective for controlling
weight gain or promoting weight loss and biologically ineffective
for promoting the synthesis of sex hormones. Steroids believed to
provide the desired weight control/weight loss characteristics include:
.DELTA.5-Androstene-3.beta.,7.alpha.-diol-17-one .DELTA.5-Androstene-3.beta.-ol-7,17-dione
.DELTA.5-Androstene-3.beta.,7.alpha.,17-triol .DELTA.5-Androstene-3.beta.,17.beta.-diol-7-one
and various derivatives thereof.
Weight loss claims
We claim:
1. A method of preventatively treating weight gain which comprises
administering to a subject in need of such treatment an effective
amount of a steroid selected from the group consisting
.DELTA.5-Androstene 3.beta.,7.alpha.,17.beta.-triol,
and
.DELTA.5-Androstene 3.beta.,17.beta.-diol-7-one,
and derivatives thereof wherein at least one of the hydroxyl groups
is esterified with an acid selected from the group consisting of
(i) a C.sub.2 to C.sub.22 aliphatic acid, (ii) a C.sub.7-12 aromatic
acid (iii) a C.sub.3 or greater dicarboxylic acid in which only
one of the carboxyl groups is esterified to the hydroxyl group(s)
on the steroid; and (iv) an inorganic acid.
2. The treatment method of claim 1 wherein the step of treating
a subject comprises the step of treating a mammal.
3. The treatment method of claim 2 wherein the step of treating
a mammal comprises the step of treating a human.
4. The treatment method of claim 1 wherein the step of treating
a subject with an effective weight gain preventative amount of a
steroid comprises the step of treating the subject with about 0.1
to about 2 grams of the steroid per 100 kg body weight per day.
5. The treatment method of claim 1 wherein the step of treating
a subject with an effective weight gain preventative amount of a
steroid comprises the step of treating the subject with about 0.5
to 2 grams of the steroid per 100 kg body weight per day.
6. The treatment method of claim 3 wherein the step of treating
a human subject with an effective weight gain preventative amount
of a steroid comprises the step of treating the human subject with
about 0.1 to 2 grams of the steroid per 100 kg body weight per day.
7. The treatment method of claim 4 wherein the step of treating
the subject with a weight gain preventative steroid comprises the
step of administering a therapeutic dose of the steroid to the subject
at least once a week.
8. The treatment method of claim 4 wherein the step of treating
the subject with a weight gain preventative steroid comprises the
step of administering a therapeutic dose of the steroid to the subject
at least once a day.
9. The treatment method of claim 1 wherein the step of treating
the subject with a weight gain preventative steroid comprises the
step of injecting the subject with the steroid.
10. The treatment method of claim 1 wherein the step of treating
the subject with a weight gain preventative steroid comprises the
step of ingesting the steroid.
11. A method of treating excessive body fat which comprises administering
to an overweight subject an effective amount of a steroid selected
from the group consisting of
.DELTA.5-Androstene 3.beta.,7.alpha.,17.beta.-triol,
and
.DELTA.5-Androstene 3.beta.,17.beta.-diol-7-one,
and derivatives thereof wherein at least one of the hydroxyl groups
is esterified with an acid selected from the group consisting of
(i) a C.sub.2 to C.sub.22 aliphatic acid, (ii) a C.sub.7-12 aromatic
acid (iii) a C.sub.3 or greater dicarboxylic acid in which only
one of the carboxyl groups is esterified to the hydroxyl group(s)
on the steroid; and (iv) an inorganic acid.
12. A method of treating obesity which comprises administering
to an obese subject a therapeutic amount of a steroid selected from
the group consisting of
.DELTA.5-Androstene 3.beta.,7.alpha.,17.beta.-triol,
and
.DELTA.5-Androstene 3.beta.,17.beta.-diol-7-one,
and derivatives thereof wherein at least one of the hydroxyl groups
is esterified with an acid selected from the group consisting of
(i) a C.sub.2 to C.sub.22 aliphatic acid, (ii) a C.sub.7-12 aromatic
acid (iii) a C.sub.3 or greater dicarboxylic acid in which only
one of the carboxyl groups is esterified to the hydroxyl group(s)
on the steroid; and (iv) an inorganic acid.
13. A biologically active steroid effective for inhibiting weight
gain without inducing a suppression of the appetite or promoting
the synthesis of sex hormones comprising .DELTA.5-Androstene 3.beta.,7.alpha.,17.beta.-triol
and derivatives thereof wherein at least one of the hydroxyl groups
is esterified with an acid selected from the group consisting of
(i) a C.sub.2 to C.sub.22 aliphatic acid, (ii) a C.sub.7-12 aromatic
acid (iii) a C.sub.3 or greater dicarboxylic acid in which only
one of the carboxyl groups is esterified to the hydroxyl group(s)
on the steroid; and (iv) an inorganic acid.
14. A biologically active steroid effective for inhibiting weight
gain without inducing a suppression of the appetite or promoting
the synthesis of sex hormones comprising .DELTA.5-Androstene 3.beta.,17.beta.-diol-7-one,
and derivatives thereof wherein at least one of the hydroxyl groups
is esterified with an acid selected from the group consisting of
(i) a C.sub.2 to C.sub.22 aliphatic acid, (ii) a C.sub.7-12 aromatic
acid (iii) a C.sub.3 or greater dicarboxylic acid in which only
one of the carboxyl groups is esterified to the hydroxyl group(s)
on the steroid; and (iv) an inorganic acid.
15. A method of preventatively treating weight gain which comprises
administering to a subject in need of such treatment an effective
amount of a steroid selected from the group consisting of
.DELTA.5-Androstene 3.beta.-ol-7,17-dione
and
.DELTA.5-Androstene 3.beta.,7-diol-17-one
and derivatives thereof wherein at least one of the hydroxyl groups
is esterified with an acid selected from the group consisting of
(i) a C.sub.2 to C.sub.22 aliphatic acid, (ii) a C.sub.7-12 aromatic
acid (iii) a C.sub.3 or greater dicarboxylic acid in which only
one of the carboxyl groups is esterified to the hydroxyl group(s)
on the steroid; and (iv) an inorganic acid.
16. A method of treating excessive body fat which comprises administering
to an overweight subject an effective amount of a steroid selected
from the group consisting of
.DELTA.5-Androstene 3.beta.-ol-7,17-dione
and
.DELTA.5-Androstene 3.beta.,7-diol-17-one
and derivatives thereof wherein at least one of the hydroxyl groups
is esterified with an acid selected from the group consisting of
(i) a C.sub.2 to C.sub.22 aliphatic acid, (ii) a C.sub.7-12 aromatic
acid (iii) a C.sub.3 or greater dicarboxylic acid in which only
one of the carboxyl groups is esterified to the hydroxyl group(s)
on the steroid; and (iv) an inorganic acid.
17. A method of treating obesity which comprises administering
to an obese subject a therapeutic amount of a steroid selected from
the group consisting of
.DELTA.5-Androstene 3.beta.-ol-7,17-dione
and
.DELTA.5-Androstene 3.beta.,7-diol-17-one
and derivatives thereof wherein at least one of the hydroxyl groups
is esterified with an acid selected from the group consisting of
(i) a C.sub.2 to C.sub.22 aliphatic acid, (ii) a C.sub.7-12 aromatic
acid (iii) a C.sub.3 or greater dicarboxylic acid in which only
one of the carboxyl groups is esterified to the hydroxyl group(s)
on the steroid; and (iv) an inorganic acid.
Weight loss description
FIELD OF THE INVENTION
Broadly, the invention relates to the use of steroids for effecting
a desired biological response. Specifically, the invention relates
to the use of a substituted dehydroepiandrosterone capable of effecting
a variety of beneficial biological responses without inducing the
formation of androgen and estrogen hormones which is commonly associated
with dehydroepiandrosterone treatment.
BACKGROUND
Dehydroepiandrosterone (.DELTA.5-androstene 3.beta.-hydroxy, 17-one)
(hereinafter referenced as DHEA) is a natural steroid produced in
the adrenal glands, testes and brain. Dehydroepiandrosterone is
an intermediate in the biosynthetic production of estrogen and androgen
(sex hormones) from 17.alpha.-hydroxy pregnenolone.
Treatment with DHEA is believed to stimulate various biological
responses including promoting weight loss and inducing an increase
in the production of the sex hormones androgen and estrogen.
The ability of DHEA to promote weight control is believed to be
mediated through enhanced thermogenesis (conversion to heat energy
rather than chemical energy such as ATP and/or triacylglycerides).
The thermogenic effect of DHEA is believed to result from a stimulation
in the synthesis of liver thermogenic enzymes such as mitochondrial
glycerol 3-phosphate dehydrogenase (G3P-DH) and cytosolic malic
enzyme (ME) which tend to reduce the efficiency of energy metabolism.
Unfortunately, DHEA is not useful as a therapeutic agent for controlling
weight gain/promoting weight loss because the dose rate of DHEA
necessary to achieve these desired characteristics may also stimulate
the production of sex hormones which is associated with various
undesired side effects.
Accordingly, a therapeutic agent possessing the weight loss characteristic
of DHEA without the associated sex hormone stimulating characteristic
would be extremely useful.
SUMMARY OF THE INVENTION
A method for controlling weight gain and/or promoting weight loss
which includes the step of treating a weight loss promoting amount
of a substituted .DELTA.5-Androstene effective for stimulating the
desired biological response while ineffective for inducing the synthesis
of sex hormones.
Steroids believed to provide the desired beneficial biological
results include:
.DELTA.5-Androstene-3.beta.,7.alpha.-diol-17-one
.DELTA.5-Androstene-3.beta.-ol-7,17-dione
.DELTA.5-Androstene-3.beta.,7.alpha.,17-triol
.DELTA.5-Androstene-3.beta.,17.beta.-diol-7-one
and derivatives thereof wherein (i) at least one of the hydroxyl
groups is esterified with an acid
selected from the group consisting of (i) C.sub.2-22 aliphatic
acids that may or may not contain one or more double bonds and may
or may not contain branched carbon chains, (ii) C.sub.7-12 aromatic
acids, (iii) C.sub.3 or larger dicarboxylic acids in which only
one of the carboxyl groups is esterified to the hydroxyl group(s)
on the steroid leaving the second carboxyl group free or in the
form of a salt, or (iv) inorganic acids such as sulfuric and phosphoric.
These steroids may also be administered as carbamates, enanthates
and other derivatives capable of releasing the free steroid in the
intestinal tract, the blood or in tissues. The desired biological
activity is a function of the steroid moiety. Derivation of the
moiety may serve a variety of possible functions including stabilization
of the steroid, flavoring or obscuring the natural flavor of the
steroid, or affecting the rate of absorption of the steroid.
DETAILED DESCRIPTION OF THE INVENTION INCLUDING A BEST MODE
.DELTA.5-Androstenes substituted at C-3, C-7 and/or C-17 with a
hydroxyl or keto group are biologically effective for controlling
weight gain and promoting weight loss without substantial stimulation
in the production of sex hormones. Derivatives of these substituted
.DELTA.5-Androstenes in which at least one of the hydroxyl groups
is esterified with an acid selected from the group consisting of
(i) C.sub.2-22 aliphatic acids that may or may not contain one or
more double bonds and may or may not contain branched carbon chains,
(ii) C.sub.7-12 aromatic acids, (iii) C.sub.3 or larger dicarboxylic
acids in which only one of the carboxyl groups is esterified to
the hydroxyl group(s) on the steroid leaving the second carboxyl
group free or in the form of a salt, or (iv) inorganic acids such
as sulfuric and phosphoric, are also believed to possess the desired
characteristics.
These steroids may also be administered as carbamates, enanthates
and other derivatives capable of releasing the free steroid in the
intestinal tract, the blood or in tissues. The desired biological
activity is a function of the steroid moiety; the derivatizing moiety
may serve to stabilize the steroid, to favor or to retard absorption
or to obscure its flavor.
SYNTHESIS
.DELTA.5-Androstene 3.beta.,7-diol, 17-one (7-hydroxy DHEA)
.DELTA.5-Androstene 3.beta.,7.alpha.-diol, 17-one (7-hydroxy DHEA)
can be synthesized from commercially available DHEA acetate by sequentially
synthesizing:
.DELTA.5-androstene 3.beta.-hydroxy-17-one acetate
.DELTA.5-androstene 3.beta.-hydroxy-7-bromo-17-one
.DELTA.5-androstene 3.beta.,7.alpha.-hydroxy-17-one diacetate
.DELTA.5-androstene 3.beta.,7.alpha.-hydroxy-17-one
.DELTA.5-androstene 3.beta.-hydroxy-7-bromo-17-one (7-bromo DHEA)
can be synthesized from .DELTA.5-androstene 3.beta.-hydroxy-17-one
acetate (DHEA acetate) by reacting the DHEA acetate with a brominating
agent such as Dibromantin (1,3 dibromo 5,5 dimethylhydantoin) or
N-bromo succinimide. 7-bromo DHEA is unstable and must be used immediately
in the next step of the process.
The 7-bromo DHEA containing an isomeric mixture of 7.alpha.-bromo
DHEA and 7.beta.-bromo DHEA may be equilibrated to 7.alpha.-bromo
DHEA in accordance with the method described for a cholesterol derivative
in Confalone, P. N., Kulesha, I. D., and Uskokovic, M. R. Jour.
Org. Chem., vol. 46, pp 1030-1032 (1981). Briefly the racemic mixture
of 7-bromo DHEA is contacted with cold anhydrous LiBr and shielded
from light until the stereospecific composition is achieved.
.DELTA.5-androstene 3.beta.,7-hydroxy-17-one diacetate (7-hydroxy
DHEA diacetate) may be synthesized from the 7-bromo DHEA by reacting
the 7-bromo DHEA with a mixture of glacial acetic acid and powdered
silver acetate at room temperature in a suitable solvent such as
methylene chloride or acetone.
.DELTA.5-androstene 3.beta.,7.alpha.-hydroxy, 17-one (7-hydroxy
DHEA)2 may be synthesized from the 7-hydroxy DHEA diacetate by reacting
the 7-hydroxy DHEA diacetate dissolved in methanol with an aqueous
solution containing a suitable base such as Na.sub.2 CO.sub.3.
The synthesized 7-hydroxy DHEA may then be purified by (i) evaporating
the methanol in vacuo, (ii) extracting the 7-hydroxy DHEA into an
appropriate organic solvent such as dichloromethane, (iii) evaporating
the organic solvent in vacuo, (iv) azeotropically drying the extracted
solids containing the 7-hydroxy DHEA with a suitable organic solvent
such as ethanol, (v) dissolving the extracted solids in acetone,
and then (vi) adding a suitable precipitating agent, such as hexane,
to the acetone solution to produce purified crystals of .DELTA.5-Androstene
3.beta.,7.alpha.-diol, 17-one.
A second crop of .DELTA.5-Androstene 3.beta.,7.alpha.-diol, 17-one
crystals may be obtained by cooling the resultant solution below
room temperature.
.DELTA.5-Androstene-3.beta.-ol 7,17-dione (7-keto DHEA)
.DELTA.5-Androstene 3.beta.-ol, 7,17-dione can be synthesized from
commercially available DHEA acetate by sequentially synthesizing:
3.beta.-acetoxy-.DELTA.5-androstene-17-one
3.beta.-acetoxy-{5-androstene-7,17-one
.DELTA.5-androstene 3.beta.-hydroxy-7,17-one
3.beta.-acetoxy-.DELTA.5-androstene-7,17-one (7-one DHEA acetate)
can be synthesized from 3.beta.-acetoxy-.DELTA.5-androstene-17-one
(DHEA acetate) by reacting the DHEA acetate with the oxidizing agent
CrO.sub.3 in accordance with the procedure outlined in Fieser, L.
F., Jour. Am. Chem. Soc., vol. 75, pp 4386-4394 (1953).
.DELTA.5-androstene 3.beta.-hydroxy-7,17-dione (7-one DHEA) can
be synthesized from the 7-one acetate and purified by employing
the deesterification and purification steps set forth above with
respect to the synthesis and purification of 7-hydroxy DHEA from
7-hydroxy DHEA diacetate.
.DELTA.5-Androstene 3.beta.,7.alpha.,17.beta.-triol (7.alpha.-hydroxy
Androstenediol)
.DELTA.5-Androstene 3.beta.,7.alpha.,17.beta.-triol can be synthesized
from commercially available Androstenediol diacetate by sequentially
synthesizing:
.DELTA.5-androstene 3.beta.,17.beta.-diol, diacetate
.DELTA.5-androstene 3.beta.,17.beta.-diol-7-bromide diacetate
.DELTA.5-androstene 3.beta.,7.alpha.,17.beta.-triol-3,17-diacetate
.DELTA.5-androstene 3.beta.,7.alpha.,17.beta.-triol
.DELTA.5-androstene 3.beta.,17.beta.-diol-7-bromide (7-bromo Androstenediol)
can be synthesized from the commercially available .DELTA.5-androstene
3.beta.,17.beta.-diol diacetate (Androstenediol diacetate) by reacting
the Androstenediol diacetate with a brominating agent such as Dibromantin
(1,3 dibromo 5,5 dimethylhydantion) or N-bromosuccinimide. The synthesized
7-bromo Androstenediol is unstable and must be used immediately.
The 7-bromo Androstenediol contains an isomeric mixture of 7.alpha.-bromo
Androstenediol and 7.beta.-bromo Androstenediol which can be equilibrated
to 7.alpha.-bromo Androstenediol in accordance with the method described
in Confalone, P. N., Kulesha, I. D., and Uskokovic, M. R. Jour.
Org. Chem., vol. 46, pp 1030-1032 (1981). Briefly the racemic mixture
of 7-bromo Androstenediol is contacted with anhydrous LiBr and shielded
from light until the stereospecific composition is achieved.
.DELTA.5-androstene 3.beta.,7.alpha.,17.beta.-triol-3,17-diacetate
(7-hydroxy Androstenediol diacetate) may be synthesized from the
7-bromo Androstenediol by reacting the 7-bromo Androstenediol with
a mixture of glacial acetic acid and silver acetate in a suitable
solvent such as methylene chloride or acetone.
.DELTA.5-androstene 3.beta.,7.alpha.,17.beta.-triol (7-hydroxy
Androstenediol) may be synthesized from the 7-hydroxy Androstenediol
diacetate by reacting the 7-hydroxy Androstenediol diacetate in
methanol with an aqueous solution containing a suitable base such
as Na.sub.2 CO.sub.3.
The synthesized 7-hydroxy Androstenediol may then be purified by
(i) evaporating the methanol in vacuo, (ii) extracting the 7-hydroxy
Androstenediol into an appropriate organic solvent such as dichloromethane,
(iii) evaporating the organic solvent in vacuo, (iv) azeotropically
drying the extracted solids containing the 7-hydroxy Androstenediol
with a suitable organic solvent such as ethanol, (v) dissolving
the extracted solids in acetone, and then (vi) adding a suitable
precipitating agent, such as hexane, to the acetone solution to
produce purified crystals of .DELTA.5-Androstene 3.beta.,7.alpha.,17.beta.-triol.
A second crop of .DELTA.5-Androstene 3.beta.,7.alpha.,17.beta.-triol
crystals may be obtained by cooling the resultant solution below
room temperature.
.DELTA.5-Androstene 3.beta.,17.beta.diol, 7-one (7-keto Androstenediol)
.DELTA.5-Androstene 3.beta.,17.beta.-diol-7-one can be synthesized
from commercially available androstenediol diacetate by sequentially
synthesizing:
.DELTA.5-androstene 3.beta.,17.beta.-diol diacetate
.DELTA.5-androstene 3.beta.,17.beta.-diol-7-one diacetate
.DELTA.5-androstene 3.beta.,17.beta.-diol-7-one
.DELTA.5-androstene 3.beta.,17.beta.-diol-7-one diacetate can be
synthesized from .DELTA.5-androstene 3.beta.,17.beta.-diol diacetate
(Androstenediol diacetate) by reacting the androstenediol diacetate
with the oxidizing agent CrO.sub.3 in accordance with the procedure
outlined in Fieser, L. F., Jour. Am. Chem. Soc., vol. 75, pp 4386-4394
(1953).
.DELTA.5-androstene 3.beta.,17.beta.-diol-7-one (7-one androstenediol)
can be synthesized from the .DELTA.5-androstene 3.beta.,17.beta.-diol-7-one
diacetate and purified by employing the deesterification and purification
steps set forth above with respect t the synthesis and purification
of 7-hydroxy DHEA from 7-hydroxy DHEA diacetate.
Without intending to be unduly limited thereby, it is believed
that the substituted .DELTA.5-Androstenes may be further modified
by esterifying one or more of the hydroxyl groups with any of a
variety of organic acids and inorganic acids such as sulfuric or
phosphoric acid.
TREATMENT
A subject may be treated with the substituted .DELTA.5-Androstenes
by any of the commonly accepted practices including orally or by
injection. It is believed that treatment at a dosage rate of about
0.1 to 2 grams, preferably about 0.5 to 2 grams, steroid per 100
kilograms body weight per day is generally effective for promoting
weight loss and/or preventing weight gain. A dose rate of less than
about 0.1 gram per 100 kilograms bodyweight is believed to be generally
ineffective for preventing weight gain while a dose rate of greater
than about 2 grams per 100 kilograms bodyweight increases the cost
of treatment without providing a corresponding benefit in performance.
The optimum dose rate to be administered to a subject is case specific
as the optimum dose rate depends upon several factors including
current body composition (percent fat), the desired effect (weight
gain prevention versus weight loss), eating habits of the individual
(daily caloric intake), and the like. As would be expected, the
dose rate provided to a subject for the purpose of promoting weight
loss will be greater than that necessary to promote weight maintenance
assuming identical caloric intake under each program.
Without intending to be limited thereby, we believe that the substituted
.DELTA.5-Androstenes are metabolic intermediates between the conversion
of DHEA to a metabolite(s) actually responsible for enhancing the
production of thermogenic enzymes such as glycerol 3-phosphate dehydrogenase
and malic enzyme.
The subject may be treated with a steroid on any desired schedule.
It is anticipated that the steroid will be effective for preventing
weight gain and/or promoting weight loss not only while actively
present within the body, but also for as long as the concentration
of the induced thermogenic enzyme(s) remain elevated. At the present
time, the duration of effectiveness for the steroid is not fully
appreciated. However, it is believed that the steroid is not stored
within the body and will be substantially removed and/or deactivated
within days after administration. Accordingly, the subject should
be conveniently treated every day for optimum performance but may
be treated less frequently such as every other day or every week
when less than maximum performance is acceptable. For example, a
subject placed on a weight maintenance program may require treatment
with the steroid only once a week even though the steroid and the
induced thermogenic enzyme(s) are not retained during the entire
period between treatments as the weight loss occurring within the
first few days after treatment counterbalances any weight gain occurring
during the remaining days between treatments.
As is apparent from the factors which affect dosage and dose rate,
each particular subject should be carefully and frequently reviewed
and the dosage and/or dose rate altered in accordance with the particular
situation.
EXPERIMENTAL
EXAMPLE I
Synthesis
.DELTA.5-Androstene 3.beta.,7.alpha.-diol-17-one
(Step 1) Into a two liter, triple neck, round bottom, flask equipped
with a magnetic stirrer and a reflux condenser was placed 1000 ml
hexane (b.p 69.degree.-71.degree. ), 10 grams (0.03 moles) DHEA
acetate and 13.6 grams (0.16 moles) NaHCO.sub.3 to form a first
solution. The first solution was placed under a N.sub.2 atmosphere
and heated under constant agitation to reflux. Into the refluxing
first solution was added 6.11 grams (0.021 moles) Dibromantin (1,3
dibromo 5,5 dimethylhydantion) as a brominating agent to form a
second solution. The second solution gradually turned orange after
which it rapidly turned a pale white/light yellow. The second solution
was refluxed for 30 minutes, cooled to room temperature and filtered
through a sintered glass funnel. The residue was rinsed with 50
ml dichloromethane and the combined filtrate rotovapped to dryness
at a temperature of less than 35.degree. C. The dry filtrate (.DELTA.5-Androstene
3.beta.-ol-7-bromo-17-one) is unstable to storage and was used immediately
in step two.
(Step 2) The dry filtrate was resolubilized in 80 ml of dichloromethane
in a one liter stoppered flask equipped with a magnetic stirrer
and placed in an ice bath. Into the resolubilized filtrate was added
8 grams anhydrous LiBr in 320 ml ice-cold acetone to form a third
solution. The third solution was shielded from light and stirred
continuously for three hours. The resulting solution of predominantly
(.DELTA.5-Androstene 3.beta.-ol-7.alpha.-bromo-17-one) was allowed
to warm briefly and used immediately in step three.
(Step 3) Into a 500 ml flask equipped with a magnetic stirrer was
placed 320 ml dichloromethane, 80 ml glacial acetic acid, and 26
grams of silver acetate to form a first suspension. The first suspension
was stirred continuously for 20 minutes at room temperature. The
stirred first suspension was added under constant agitation into
the warmed solution of predominantly .DELTA.5-Androstene 3.beta.-ol-7.alpha.-bromo-17-one
to form a second suspension. The second suspension was constantly
stirred for 30 minutes at room temperature during which the suspension
gradually darkened. The darkened suspension was filtered through
a sintered glass funnel to produce a solid fraction. The filtered
solid fraction was rinsed with 100 ml dichloromethane. The filtrate
was extracted three times with 1000 ml of water, neutralized with
1000 ml of a 5% NaHCO.sub.3 solution, and extracted twice more with
water. The organic solution containing the .DELTA.5-Androstene 3.beta.,17.beta.-diol-17-one
diacetate was then rotovapped to dryness.
(Step 4) The dried extracted solids were resolubilized in 500 ml
methanol in a one liter, triple necked flask equipped with a magnetic
stirrer and a reflux condenser to form a fourth solution. The fourth
solution was placed under a N.sub.2 atmosphere and heated under
constant stirring to reflux. Into the fourth solution was added
250 ml of a 5% aqueous solution of Na.sub.2 CO.sub.3 to form a fifth
solution. The fifth solution was refluxed under constant agitation
for 45 minutes. The methanol was rotovapped off and the aqueous
fifth solution carefully brought to a pH of 7 with an appropriate
amount of glacial acetic acid. The neutralized fifth solution was
extracted twice with 100 ml of dichloromethane. The dichloromethane
solution of .DELTA.5-Androstene 3.beta.,7.alpha.-diol-17-one was
rotovapped to near dryness, azeotropically dried with absolute ethanol,
and then azeotropically dried twice with acetone. Warm acetone was
added to the dried extracted solids until the solids were completely
dissolved to form a sixth solution. Hexane was added to the sixth
solution until the solution began to cloud at which time crystals
of .DELTA.5-Androstene 3.beta.,7.alpha.-diol-17-one began to form
at room temperature.
A second crop of .DELTA.5-Androstene 3.beta.,7.alpha.-diol-17-one
crystals was obtained by cooling the remaining sixth solution.
The product melts at about 187.degree.-189.degree. C. and when
recrystallized from acetone/hexane melts at about 192.degree.-193.degree.
C.
Example II
Synthesis
.DELTA.5-Androstene 3.beta.,7(.alpha.,.beta.)-diol-17-one
.DELTA.5-Androstene 3.beta.,7(.alpha.,.beta.)-diol-17-one was manufactured
in accordance with the procedure set forth in Example I except that
Step 2 was eliminated with the dried filtrated from Step 1 simply
resolubilized in the 80 ml of dichloromethane in preparation for
Step 3.
Example III
Synthesis
.DELTA.5-Androstene-3.beta.-ol-7,17-dione
(Step 1) Into a 50 ml flask equipped with a magnetic stirrer and
a water bath was placed 6.5 ml acetic anhydride, 23 ml acetic acid,
1.7 grams sodium acetate, and 2 grams DHEA acetate to form a first
mixture. Into the first mixture was added 2 grams chromium trioxide
over a thirty minute period to form a second mixture. The first
mixture was maintained at a constant temperature of 56.degree.-58.degree.
C. and continuously agitated during addition of the chromium trioxide.
The second mixture was maintained at 56.degree.-58.degree. C. and
continuously agitated for an additional hour after which the second
mixture was cooled and slowly poured under continuous agitation
into 600 ml of ice water to form a precipitate. The flocculent precipitate
was collected on a sintered glass funnel and washed with water until
no longer green. After drying in vacuo over P.sub.2 O.sub.5 the
product was dissolved in methanol and then recrystallized to yield
substantially pure .DELTA.5-Androstene 3.beta.-acetoxy-7,17-dione
having a melting point of about 191.degree.-192.degree. C.
(Step 2) The precipitate was resolubilized in 500 ml of methanol
in a one liter, triple necked, round bottom flask equipped with
a magnetic stirrer and a reflux condenser to form a third solution.
The third solution was placed under a N.sub.2 atmosphere and heated
under constant agitation to reflux. Into the third solution was
added 250 ml of a 5% aqueous solution of Na.sub.2 CO.sub.3 to form
a fourth solution. The fourth solution was refluxed under constant
agitation for 45 minutes. The methanol was rotovapped off and the
aqueous fourth solution carefully brought to a pH of 7 with an appropriate
amount of glacial acetic acid. The neutralized fourth solution was
extracted with two 100 ml portions of dichloromethane, the two portions
combined, and the dichloromethane evaporated in vacuo. The extracted
solids were then azeotropically dried first with absolute ethanol
and then with two separate portions of acetone. Methanol was added
to the dried extracted solids until the solids were completely dissolved
to form a fifth solution. Hexane was added to the fifth solution
until the solution began to cloud at which time crystals of .DELTA.5-Androstene-3.beta.-ol-7,17-dione
began to form at room temperature.
A second crop of .DELTA.5-Androstene-3.beta.-ol-7,17-dione crystals
was obtained by cooling the remaining sixth solution.
The resultant product had a melting point of about 35.degree.-238.degree.
C.
Example IV
Synthesis
.DELTA.5-Androstene 3.beta.,7,17-triol
(Step 1) Into a two liter round bottom flask equipped with a magnetic
stirrer and a reflux condenser was placed 1000 ml hexane (b.p 69.degree.-71.degree.
), 10 grams (0.03 moles) .DELTA.5-Androstene-3.beta.,17.beta.-diol
diacetate and 13.6 grams (0.16 moles) NaHCO.sub.3 to form a first
solution. The first solution was placed under a N.sub.2 atmosphere
and heated under constant agitation to reflux. Into the refluxing
first solution was added 6.11 g (0.021 moles) Dibromantin (1,3-dibromo-5,5-dimethylhydantion)
as a brominating agent to form a second solution. The second solution
gradually turned orange after which it rapidly turned a pale white/light
yellow. The second solution was refluxed for 30 minutes, cooled
to room temperature and filtered through a sintered glass funnel.
The residue was rinsed with 50 ml dichloromethane and the filtrates
rotovapped to dryness at a temperature of less than 35.degree. C.
The dry filtrate (.DELTA.5-Androstene-3.beta.,17-diol-7-bromide)
is unstable to storage and was used immediately in step two.
(Step 2) The dried filtrate was resolubilized in 80 ml of dichloromethane
in a flask equipped with a magnetic stirrer and placed in an ice
bath. Into the resolubilized filtrate was added 8 g anhydrous LiBr
in 320 ml ice-cold acetone to form a third solution. The third solution
was shielded from light and stirred continuously for three hours.
The resulting solution of predominantly .DELTA.5-Androstene-3.beta.,17.beta.-diol-7.alpha.-bromide
was allowed to warm briefly and used immediately.
(Step 3) Into a 500 ml flask equipped with a magnetic stirrer was
placed 320 ml methylene chloride, 80 ml glacial acetic acid, and
26 grams silver acetate to form a first suspension. The first suspension
was stirred continuously for 20 minutes at room temperature. The
stirred first suspension was added under constant agitation to the
warmed solution of predominantly .DELTA.5-Androstene-3.beta.,17-diol-7.alpha.-bromide
to form a second suspension. The second suspension was constantly
stirred for 30 minutes at room temperature during which the suspension
gradually darkened and was then filtered through a sintered glass
funnel. The residual solids retained on the glass filter were rinsed
with 100 ml dichloromethane. The filtrate was extracted three times
with 1000 ml of water, neutralized with 1000 ml of a 5% NaHCO.sub.3
solution, and then extracted twice more with water. The solids extracted
from the filtrate (.DELTA.5-Androstene-3.beta.,7.alpha.,17.beta.-triol-3,17-diacetate)
were rotovapped to dryness.
(Step 4) The dried extracted solids were resolubilized in 500 ml
methanol contained in a one liter, triple necked, round bottom flask
equipped with a magnetic stirrer and a reflux condenser to form
a fourth solution. The fourth solution was placed under a N.sub.2
atmosphere and heated under constant agitation to reflux. Into the
fourth solution was added 250 ml of a 5% aqueous solution of Na.sub.2
CO.sub.3 to form a fifth solution. The fifth solution was refluxed
under constant agitation for 45 minutes. The methanol was rotovapped
off and the aqueous fifth solution carefully brought to a pH of
7 with an appropriate amount of glacial acetic acid. The neutralized
fifth solution was extracted twice with 100 ml dichloromethane and
the combined extract evaporated in vacuo. The extracted solids (.DELTA.5-Androstene-3.beta.,7.alpha.,17.beta.-triol)
were azeotropically dried with absolute ethanol and then twice with
acetone. Warm acetone was added to the dried extracted solids until
the solids were completely dissolved to form a sixth solution. Hexane
was added to the sixth solution until the solution began to cloud
at which time crystals of .DELTA.5-Androstene-3.beta.,7.alpha.,17.beta.-triol
began to form at room temperature.
A second crop of .DELTA.5-Androstene-3.beta.,7.alpha.,17.beta.-triol
crystals was obtained by cooling the remaining sixth solution.
Example V
Synthesis
.DELTA.5-Androstene-3.beta.,7(.alpha.,.beta.),17-triol
.DELTA.5-Androstene-3.beta.,7(.alpha.,.beta.),17-triol was manufactured
in accordance with the procedure set forth in Example III except
that Step 2 was eliminated and the dried filtrated material from
Step 1 was simply resolubilized in 80 ml of methylene chloride in
preparation for Step 3.
Example VI
Synthesis
.DELTA.5-Androstene-3.beta.,17.beta.-diol-7-one
(Step 1) Into a 50 ml flask equipped with a magnetic stirrer and
a water bath was placed 6.5 ml acetic anhydride, 23 ml acetic acid,
1.7 grams sodium acetate, and 2 grams androstenediol diacetate to
form a first mixture. Into the first mixture was added 2 grams chromium
trioxide over a thirty minute period to form a second mixture. The
first mixture was maintained at a constant temperature of 56.degree.-58.degree.
C. and continuously agitated during addition of the chromium trioxide.
The second mixture was maintained at 56.degree.-58.degree. C. and
continuously agitated for an additional hour after which the second
mixture was cooled and slowly poured under continuous agitation
into 600 ml of ice water to form a precipitate. The flocculent precipitate
was filtered through a sintered glass funnel, washed with water
until no longer green and dried in vacuo.
(Step 2) The dried precipitate was resolubilized in 500 ml of methanol
contained in a one liter, round bottom flask equipped with a magnetic
stirrer and a reflux condenser to form a third solution. The third
solution was placed under a N.sub.2 atmosphere and heated under
constant agitation to reflux. Into the third solution was added
250 ml of a 5% aqueous solution of Na.sub.2 CO.sub.3 to form a fourth
solution. The fourth solution was refluxed under constant agitation
for 45 minutes. The methanol was rotovapped off and the aqueous
fourth solution carefully brought to a pH of 7 with an appropriate
amount of glacial acetic acid. The neutralized fourth solution was
extracted twice with 100 ml portions of dichloromethane and the
combined extract evaporated in vacuo. The extracted solids were
azeotropically dried with absolute ethanol and then twice with acetone.
Methanol was added to the dried extracted solids until the solids
were completely dissolved to form a fifth solution. Hexane was added
to the fifth solution until the solution began to cloud at which
time crystals of .DELTA.5-Androstene-3.beta.,17.beta.-diol-7-one
began to form at room temperature.
The resultant product had a melting point of about 200.degree.-202.degree.
C.
|