U.S. patent application number 11/040964 was filed with the patent office on 2005-06-16 for method of making and using 7alpha,11beta-dimethyl-17beta-hydroxyestr-4-en-- 3-one 17-undecanoate.
This patent application is currently assigned to Government of the USA, represented by the Secretary, Department of Health and Human Services, Government of the USA, represented by the Secretary, Department of Health and Human Services. Invention is credited to Blye, Richard P., Kim, Hyun K..
Application Number | 20050130944 11/040964 |
Document ID | / |
Family ID | 22990907 |
Filed Date | 2005-06-16 |
United States Patent
Application |
20050130944 |
Kind Code |
A1 |
Blye, Richard P. ; et
al. |
June 16, 2005 |
Method of making and using
7alpha,11beta-dimethyl-17beta-hydroxyestr-4-en-- 3-one
17-undecanoate
Abstract
Disclosed are methods of using
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxy- estr-4-en-3-one
17-undecanoate (II) 1 for various hormonal therapies, dosage forms
comprising
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxyestr-4-en-3-one
17-undecanoate, and processes for its preparation.
Inventors: |
Blye, Richard P.; (Highland,
MD) ; Kim, Hyun K.; (Bethesda, MD) |
Correspondence
Address: |
LEYDIG, VOIT & MAYER, LTD.
TWO PRUDENTIAL PLAZA, SUITE 4900
180 NORTH STETSON AVENUE
CHICAGO
IL
60601-6780
US
|
Assignee: |
Government of the USA, represented
by the Secretary, Department of Health and Human Services
Rockville
MD
|
Family ID: |
22990907 |
Appl. No.: |
11/040964 |
Filed: |
January 21, 2005 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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11040964 |
Jan 21, 2005 |
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10260854 |
Sep 30, 2002 |
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10260854 |
Sep 30, 2002 |
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PCT/US01/10293 |
Mar 30, 2001 |
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60193530 |
Mar 31, 2000 |
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60194440 |
Apr 4, 2000 |
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Current U.S.
Class: |
514/170 ;
514/178 |
Current CPC
Class: |
C07J 1/007 20130101;
A61P 15/16 20180101; C07J 1/0059 20130101; C07J 71/001 20130101;
A61P 5/24 20180101; C07J 1/0074 20130101; C07J 21/006 20130101;
A61K 31/56 20130101 |
Class at
Publication: |
514/170 ;
514/178 |
International
Class: |
A61K 031/57; A61K
031/56 |
Claims
1-38. (canceled)
39. A crystalline compound of formula 10 12having a melting point
of 155-157.degree. C.
40. A compound of formula 5 13
41. A compound of formula 6 14
42. A compound of formula 7 15
43. A compound of formula 9 16
44-45. (canceled)
46. A method for providing hormonal therapy to a patient comprising
the oral administration of up to about 75 mg/day of
7.alpha.,11.beta.-dimethy- l-17.beta.-hydroxyestr-4-en-3-one
17-undecanoate to a patient in need thereof.
47. The method of claim 46, wherein the hormonal therapy continues
at least 3 months.
48. The method of claim 46, wherein the hormonal therapy is the
treatment of hypogonadism in a male patient.
49. The method of claim 48, wherein up to about 50 mg/day of
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxyestr-4-en-3-one
17-undecanoate is administered to the patient.
50. The method of claim 49, wherein up to about 25 mg/day of
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxyestr-4-en-3-one
17-undecanoate is administered to the patient.
51. The method of claim 46, wherein hormonal therapy is male
contraception.
52. The method of claim 51, wherein the therapy continues for at
least 3 months.
53. The method of claim 51, wherein up to about 50 mg/day
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxyestr-4-en-3-one
17-undecanoate is administered to the patient.
54. The method of claim 51, wherein up to about 30 mg/day of
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxyestr-4-en-3-one
17-undecanoate is administered to the patient.
55. The method of claim 51, further comprising the administration
of estrogen, a progestin, or both.
56. The method of claim 46, wherein the hormone therapy is the
promotion and maintenance of muscle mass in a patient.
57. The method of claim 56, wherein the therapy continues for at
least 3 months.
58. The method of claim 46, wherein the hormonal therapy is the
pilliative treatment of breast cancer in women.
59. The method of claim 47, wherein
7.alpha.,11.beta.-dimethyl-17.beta.-hy- droxyestr-4-en-3-one
17-undecanoate is administered as a formulation comprising
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxyestr-4-en-3-one
17-undecanoate and an oily carrier.
60. A method for providing hormonal therapy to a patient comprising
administering parenterally to the patient an average dosage of up
to about 600 mg/month of
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxyestr-4-e- n-3-one
17-undecanoate during the treatment period.
61. The method of claim 60, wherein
7.alpha.,11.beta.-dimethyl-17.beta.-hy- droxyestr-4-en-3-one
17-undecanoate is parenterally administered as a formulation
comprising 7.alpha.,11.beta.-dimethyl-17.beta.-hydroxyestr-4--
en-3-one 17-undecanoate as a solid and an aqueous carrier, with the
amount of
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxyestr-4-en-3-one
17-undecanoate administered at each interval being selected so that
the average amount of
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxyestr-4-en-3-- one
17-undecanoate administered during the period of hormonal treatment
averages up to about 50 mg/week.
62. The method of claim 61, wherein
7.alpha.,11.beta.-dimethyl-17.beta.-hy- droxyestr-4-en-3-one
17-undecanoate is crystalline and the formulation comprises a
suspension of 7.alpha.,11.beta.-dimethyl-17.beta.-hydroxyestr-
-4-en-3-one 17-undecanoate in the aqueous carrier.
63. The method of claim 60, wherein
7.alpha.,11.beta.-dimethyl-17.beta.-hy- droxyestr-4-en-3-one
17-undecanoate is parenterally administered as a formulation
comprising 7.alpha.,11.beta.-dimethyl-17.beta.-hydroxyestr-4--
en-3-one 17-undecanoate and an aqueous carrier, with up to about
600 mg of
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxyestr-4-en-3-one
17-undecanoate being administered once over a period of at least
about 1 month.
64. The method of claim 60, wherein
7.alpha.,11.beta.-dimethyl-17.beta.-hy- droxyestr-4-en-3-one
17-undecanoate is parenterally administered as a formulation
comprising 7.alpha.,11.beta.-dimethyl-17.beta.-hydroxyestr-4--
en-3-one 17-undecanoate and an aqueous carrier, with up to about
450 mg of
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxyestr-4-en-3-one
17-undecanoate being administered once over a period of at least
about 2 months.
65. The method of claim 60, wherein
7.alpha.,11.beta.-dimethyl-17.beta.-hy- droxyestr-4-en-3-one
17-undecanoate is parenterally administered as a formulation
comprising 7.alpha.,11.beta.-dimethyl-17.beta.-hydroxyestr-4--
en-3-one 17-undecanoate and an aqueous carrier, with up to about
300 mg of
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxyestr-4-en-3-one
17-undecanoate being administered once over a period of at least
about 3 months.
66. The method of claim 60, wherein the hormonal treatment is the
treatment of hypogonadism in male patients.
67. The method of claim 61, wherein
7.alpha.,11.beta.-dimethyl-17.beta.-hy- droxyestr-4-en-3-one
17-undecanoate is administered in a parenteral formulation
comprising 7.alpha.,11.beta.-dimethyl-17.beta.-hydroxyestr-4--
en-3-one 17-undecanoate and an aqueous carrier.
68. The method of claim 67, wherein from about 150 mg to about 450
mg of 7.alpha.,11.beta.-dimethyl-17.beta.-hydroxyestr-4-en-3-one
17-undecanoate is patenterally administered once over a period of
at least about 1 month.
69. The method of claim 68, wherein
7.alpha.,11.beta.-dimethyl-17.beta.-hy- droxyestr-4-en-3-one
17-undecanoate is parenterally administered once over a period of
at least about 2 months.
70. The method of claim 67, wherein
7.alpha.,11.beta.-dimethyl-17.beta.-hy- droxyestr-4-en-3-one
17-undecanoate is parenterally administered once over a period of
at least about 3 months.
71. The method of claim 60, wherein the hormonal treatment is male
contraception.
72. The method of claim 71, wherein
7.alpha.,11.beta.-dimethyl-17.beta.-hy- droxyestr-4-en-3-one
17-undecanoate is parenterally administered at an average dosage of
up to about 200 mg/week.
73. The method of claim 71, further comprising the administration
of contraception-effective amounts of estrogen, progestins or
mixtures thereof, wherein
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxyestr-4-en-3-o- ne
17-undecanoate is parenterally administered up to about 100 mg at
intervals of at least about 2 weeks.
74. The method of claim 73, wherein
7.alpha.,11.beta.-dimethyl-17.beta.-hy- droxyestr-4-en-3-one
17-undecanoate is parenterally administered up to about 75 mg at
intervals of at least about 1 month.
75. An oral dosage formulation comprising
7.alpha.,11.beta.-dimethyl-17.be- ta.-hydroxyestr-4-en-3-one
17-undecanoate and a pharmaceutically-acceptabl- e carrier.
76. The oral dosage formulation of claim 75, wherein the
pharmaceutically-acceptable carrier comprises an oil.
77. The oral dosage formulation of claim 76, wherein the
formulation comprises up to about 25 mg of
7.alpha.,11.beta.-dimethyl-17.beta.-hydrox- yestr-4-en-3-one
17-undecanoate
78. The oral dosage formulation of claim 77, wherein the
formulation comprises up to about 15 mg of
7.alpha.,11.beta.-dimethyl-17.beta.-hydrox- yestr-4-en-3-one
17-undecanoate.
79. An aqueous formulation for parenteral administration comprising
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxyestr-4-en-3-one
17-undecanoate and water.
80. The aqueous formulation of claim 79, wherein
7.alpha.,11.beta.-dimethy- l-17.beta.-hydroxyestr-4-en-3-one
17-undecanoate is in crystalline form.
81. A method for preparing
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxyestr- -4-en-3-one
17-undecanoate (Compound II) comprising the steps of: (a)
converting the ether group of Compound 1 17to a carbonyl group,
providing Compound 2 18(b) ketalizing the carbonyl group of
Compound 2 to provide Compound 4; 19(c) epoxidizing Compound 4 to
provide the epoxide of Compound 5; 20(d) opening the epoxide ring
in Compound 5 and substituting an alkyl group at C.sub.11 to
provide Compound 6 (comprising a mixture of 7.alpha.- and
7.beta.-methyl isomers, Compounds 6a and 6b, respectively) by use
of a Grignard reagent; 21(e) deketalizing and dehydrating Compound
6 to provide Compound 7; 22(f) converting Compound 7a to Compound
9; 23(g) converting Compound 9 to Compound 10; and 24(h)
esterifying Compound 10 to provide Compound II 25
82. 7.alpha.,11.beta.-dimethyl-17.beta.-hydroxyestr-4-en-3-one
17-undecanoate (Compound II) in crystalline form.
83. The compound of claim 82, having a m.p. of 62-64.degree. C.
84. The compound of claim 82, wherein the purity of the crystalline
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxyestr-4-en-3-one
17-undecanoate is at least 99%.
85. The oral dosage form of claim 75, wherein the formulation
comprises up to about 75 mg of
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxyestr-4-en-3-- one
17-undecanoate.
86. The aqueous formulation of claim 79, wherein the formulation
comprises up to about 600 mg of
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxyestr-4-e- n-3-one
17-undecanoate.
Description
CROSS-REFERENCE TO RELATED PATENT APPLICATIONS
[0001] This application claims the benefit of U.S. Provisional
Patent Application No. 60/193,530, filed Mar. 30, 2000 and
60/194,440, filed Apr. 4, 2000, both of which are incorporated
herein by reference.
FIELD OF THE INVENTION
[0002] The present invention generally relates to methods of making
and using esters of androgenic steroids.
BACKGROUND OF THE INVENTION
[0003] Androgen is a term used to identify the human male sex
hormones. These hormones, which are chemically classified as
steroids, are produced in the body by the testis, the cortex of the
adrenal gland and, to a much lesser extent, by the ovaries.
Testosterone is perhaps the most widely recognized androgen, and is
responsible for the development of male characteristics in a human,
including secondary sexual characteristics, libido and the ability
to produce sperm.
[0004] When a person is unable to synthesize testosterone, therapy
directed at replacing the missing hormone is commonly undertaken.
In practice, however, this therapy can be problematic. For example,
testosterone exhibits only weak activity when administered orally.
While parenteral administration is possible, it is impractical
because testosterone remains active in the body for only a short
time. Research has therefore focused on identifying so-called
synthetic androgens that are acceptable substitutes for natural
testosterone.
[0005] A number of oral and injectable synthetic androgens have
been developed over the years, including esters of various
androgens. While these esters are hydrolyzed in the body into their
corresponding biologically-active alcohols, they are nonetheless
administered because they slow the rapid degradation of the
synthetic androgen by the body. This maximizes the amount of the
biologically active alcohol that reaches the bloodstream.
[0006] Unfortunately, the activity of these androgen esters is
unpredictable. Different androgens sharing the same ester group
exhibit varying and unpredictable levels of activity, as do
androgens having the same basic chemical structure, but different
ester groups.
[0007] One of the esters that has emerged as a viable injectable
synthetic androgen is testosterone enanthate. This enanthate is
presently used extensively via intramuscular (IM) injection for
hormone replacement therapy in hypogonadal men, and as the
androgenic component of several experimental male contraceptives.
One drawback of this active is that it is not exceptionally
long-acting--it must be administered IM every two weeks to maintain
testosterone levels within a normal (therapeutic) range in
hypogonadal men.
[0008] More specifically, testosterone enanthate is presently
administered IM for the treatment of hypogonadism at a dose of 200
mg every two or three weeks. If this enanthate is used for male
contraception, it may be administered parenterally at from about
200-400 mg every week, and if used as the androgenic component with
estrogen or progestins for contraception, it may be administered at
about 200 mg every two weeks. Testosterone bucyclate is another
synthetic androgen disclosed in, e.g., U.S. Pat. No. 4,948,790. If
administered parenterally for the treatment of hypogonadism, this
bucyclate would require a dose of about 1200 mg (given as 3
injections of 1 ml each due to its solubility) to retain activity
for about 2-3 months.
[0009] The development of androgens that exhibit activity after
oral administration has been less successful. At present, the most
widely used effective oral formulation includes methyltestosterone
as the active ingredient, administered at 10-50 mg
methyltestosterone/day. However, this active cannot be administered
on a long-term basis, as is required in androgen replacement
therapy, because of its associated liver toxicity. It is well known
that androgens alkylated at the C.sub.17 position, such as
methyltestosterone, exhibit such toxicity. While removal of the
C.sub.17 alkyl group may appear at first glance to be an obvious
solution to this problem, alkylation at this position is believed
to be necessary to prevent degradation of the active by the liver
after oral administration.
[0010] Illustrative of the development efforts relating to
synthetic androgens is U.S. Pat. No. 5,952,319. While this patent
identifies a number of potentially-active synthetic androgens,
including
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxy-4-estren-3-one
17.beta.-trans-4-n-butylcyclohexane carboxylate (referred to herein
as 7.alpha.,11.beta.-dimethyl-17.beta.-hydroxy-4-estren-3-one
bucyclate), it provides no data regarding the biological activity
of 7.alpha.,11.beta.-dimethyl-17.beta.-hydroxy-4-estren-3-one
bucyclate. There is similarly no data available concerning the
biological activity of another synthetic androgen,
7.alpha.,11.beta.-dimethyl-17.beta.-hydrox- yestr-4-en-3-one
17-undecanoate.
[0011] A need therefore exists for a means of overcoming the
foregoing and other problems associated with androgen replacement
and other therapies that require the administration of
androgens.
BRIEF SUMMARY OF THE INVENTION
[0012] The present invention meets the aforesaid and other needs by
providing, in one aspect, a method for providing hormonal therapy
to a patient comprising the oral administration of about 1 mg/day
to about 25 mg/day of
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxy-4-estren-3-one
bucyclate,
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxyestr-4-en-3-one
17-undecanoate, or a mixture thereof, to a patient in need
thereof.
[0013] This aspect of the invention is predicated in significant
part on the unexpected discoveries that
7.alpha.,11.beta.-dimethyl-17.beta.-hydro- xy-4-estren-3-one
bucyclate (also referred to herein as "the bucyclate") and
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxyestr-4-en-3-one
17-undecanoate (also referred to herein as "the undecanoate") do
not degrade after oral administration even though each lacks an
alkyl group at the C.sub.17 position, and exhibit activity far in
excess of the current oral standard, methyltestosterone. These
surprising discoveries permit hormonal therapies requiring the
administration of an androgen to be conducted utilizing oral
dosages of the bucyclate and/or the undecanoate that are
significantly lower than those required when administering oral
methyltestosterone to effect the same therapy. A further expected
benefit of using the bucyclate and undecanoate is that liver
toxicity, if any, should be minimal because these compounds are not
alkylated at the C.sub.17 position.
[0014] In another aspect, the present invention comprises a method
for providing hormonal treatment comprising the parenteral
administration of from about 1 mg up to about 100 mg of the
bucyclate and/or the undecanoate at intervals of at least about two
weeks, and preferably up to about 600 mg at much longer intervals,
e.g., a single administration of 600 mg providing effective therapy
for up to about three months.
[0015] This aspect of the invention is predicated in part on the
surprising relatively high potency, and unexpected long-term
activity, of the bucyclate and undecanoate when administered
parenterally, which potency is higher and activity longer-lasting
than esters of other potent androgenic steroids, even bucyclic
esters thereof. This activity was unexpected in view of the
preparation and evaluation of several bucyclic esters of potent
androgenic steroids other than 7.alpha.,11.beta.-dimethy-
l-17.beta.-hydroxy-4-estren-3-one bucyclate, the former group of
esters yielding disappointing results.
[0016] Another aspect of the present invention includes separate
processes for preparing the bucyclate and undecanoate, which
provide these actives in relatively high yield, and advantageously
in a solid form, preferably crystalline, at room temperature. As
both can be produced in solid form, the preparation of aqueous
microcrystalline suspensions for parenteral administration is
possible. Moreover, because these actives are solid at room
temperature, one is able to control the average particle size and
particle size distribution of the solids, thereby positively
affecting the duration of activity after parenteral administration
of the respective suspensions.
[0017] Related aspects of the present invention include certain
intermediates, in amorphous or, preferably, crystalline form, as
well as one or more steps used in the aforementioned preferred
process for preparing the bucyclate and the undecanoate.
[0018] Further aspects of the present invention include various
formulations of these two actives, including tablets, caplets,
extended release tablets, soft gelcaps containing the bucyclate
and/or undecanoate in an oily carrier, transdermal patches,
pre-filled syringes, vials and the like, in which the amount of the
bucyclate and/or undecanoate included therein may be determined in
view of their unexpected relatively high potency and long-term
activity.
[0019] It is contemplated that the hormonal therapy of the present
invention includes, but is not limited to, hormone replacement
therapy in males and females, male contraception, and the treatment
of certain cancers, such as breast cancer in females.
[0020] These and other aspects and features of the present
invention may best be understood with reference to the accompanying
figures and in the following description of the preferred
embodiments.
BRIEF DESCRIPTION OF THE DRAWINGS
[0021] FIG. 1 illustrates the chemical structure of
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxy-4-estren-3-one
bucyclate, with numerals identifying the various carbon atom
positions, including the non-alkylated C.sub.17 position.
[0022] FIG. 2A illustrates the chemical structure of
methyltestosterone.
[0023] FIG. 2B illustrates the chemical structure of testosterone
enanthate.
[0024] FIG. 3 is a graph comparing the androgenic potency of
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxy-4-estren-3-one
bucyclate and that of other compounds after oral
administration.
[0025] FIG. 4 is a graph comparing the androgenic potency of
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxy-4-estren-3-one
bucyclate and that of other compounds after oral
administration.
[0026] FIG. 5 is a graph comparing the duration of activity of
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxy-4-estren-3-one
bucyclate and that of other compounds after subcutaneous
injection.
[0027] FIG. 6 is a graph comparing the androgenic potency of
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxy-4-estren-3-one
bucyclate and that of other compounds after oral and subcutaneous
injection.
[0028] FIG. 7 is a graph comparing the androgenic potency of
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxy-4-estren-3-one
bucyclate and that of other compounds after subcutaneous
injection.
[0029] FIG. 8 is a graph comparing the duration of activity of
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxy-4-estren-3-one
bucyclate and that of other compounds after subcutaneous
injection.
[0030] FIG. 9 is a graph comparing the duration of activity of
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxy-4-estren-3-one
bucyclate and that of other compounds after subcutaneous
injection.
[0031] FIG. 10 is a graph comparing testosterone serum levels
(pg/ml) after subcutaneous injection of
7.alpha.,11.beta.-dimethyl-17.beta.-hydro- xy-4-estren-3-one
bucyclate and other compounds.
[0032] FIG. 11 is a description of a preferred method for preparing
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxy-4-estren-3-one
bucyclate.
[0033] FIG. 12 illustrates the chemical structure of
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxyestr-4-en-3-one
17-undecanoate, with numerals identifying the various carbon atom
positions, including the non-alkylated C.sub.17 position.
[0034] FIG. 13 is a graph comparing the androgenic potency of
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxyestr-4-en-3-one
17-undecanoate and testosterone after subcutaneous injection.
[0035] FIG. 14 is a graph comparing the androgenic potency of
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxyestr-4-en-3-one
17-undecanoate and methyltestosterone after oral
administration.
[0036] FIG. 15 is a graph comparing the duration of activity of
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxyestr-4-en-3-one
17-undecanoate and that of testosterone enanthate (CDB-112F) after
subcutaneous injection.
[0037] FIG. 16 is a description of a preferred method for preparing
7.alpha.,11.beta.-dimethyl-1 7.beta.-hydroxyestr-4-en-3-one
17-undecanoate.
[0038] The various aspects of the present invention described in
the following paragraphs are set forth with an emphasis on
preferred embodiments. However, it will be obvious to those of
ordinary skill in the art that variations of the preferred
embodiments may be successfully used, and that it is intended that
the invention may be practiced otherwise than as specifically
described herein. The inventive methods, processes and formulations
should therefore not be construed as being limited to the preferred
embodiments described herein.
DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS
[0039] The present invention provides a variety of methods for
providing hormonal therapy to a patient in need thereof. Each
method requires the administration of particular actives,
7.alpha.,11.beta.-dimethyl-17-hydro- xy-4-estren-3-one bucyclate,
and 7.alpha.,11.beta.-dimethyl-17.beta.-hydro- xyestr-4-en-3-one
17-undecanoate, either in combination or, preferably, alone. The
chemical structure of the bucyclate and undecanoate actives, with
numerals identifying the various carbon atom positions, are set
forth in FIGS. 1 and 12, respectively.
[0040] In significant part, the present invention rests upon the
discovery that both
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxy-4-estren-3-one
bucyclate and
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxyestr-4-en-3-one
17-undecanoate exhibit surprising and unexpected properties in
vivo. These properties permit these actives to be administered
either orally or parenterally, in relatively lower amounts, at
longer time intervals, and with less side effects, as compared to
existing alternative synthetic androgens, e.g., methyltestosterone,
testosterone enanthate. The chemical structures of these two
well-known compounds (methyltestosterone, testosterone enanthate)
are set forth in FIGS. 2A and 2B, respectively.
[0041] The surprising properties of
7.alpha.,11.beta.-dimethyl-17.beta.-hy- droxy-4-estren-3-one
bucyclate and 7.alpha.,11.beta.-dimethyl-1
7.beta.-hydroxyestr-4-en-3-one 17-undecanoate render these actives
well suited for any hormonal therapy in which an androgen is
required, or desired. By way of example only, and without intending
to limit the therapeutic uses of the actives, the actives may be
used in the treatment of hypogonadal males. The actives may also be
administered (either alone or, more effectively, in combination
with one or more steroidal progestins or estrogens) to induce and
maintain fertility suppression in male animals. Further, and due to
their anabolic properties, the actives may be administered to
promote and maintain muscle growth and maintenance. These
properties can be particularly important in persons afflicted with
muscle wasting diseases such as AIDS, but are more generally
applicable to the elderly who typically have relatively low muscle
mass. In addition, the actives may be used for the treatment of
cancer, e.g., the pilliative treatment of breast cancer in
women.
[0042] The unexpected properties of
7.alpha.,11.beta.-dimethyl-17.beta.-hy- droxy-4-estren-3-one
bucyclate and 7.alpha.,11.beta.-dimethyl-17.beta.-hyd-
roxyestr-4-en-3-one 17-undecanoate were discovered after a series
of in vivo animal studies undertaken in Sprague-Dawley rats. Based
upon these experiments, it was unexpectedly found that the
bucyclate and undecanoate, despite their lack of alkylation at the
C.sub.17 position, not only do not degrade after oral
administration, but exhibit activity far in excess of the current
oral standard, methyltestosterone. Moreover, this lack of
alkylation is expected to minimize, or eliminate, any attendant
liver toxicity. Thus, the foregoing and other therapies may be
conducted utilizing dosages of the bucyclate and undecanoate that
are significantly lower than those expected, and less than that
required when administering methyltestosterone, to effect the same
therapy. This is accomplished without the attendant concern of
liver toxicity associated with existing synthetic androgens.
[0043] More specifically, it was found that the oral activity of
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxy-4-estren-3-one
bucyclate was about four times greater than methyltestosterone. The
oral activity of the undecanoate,
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxyestr-4-en-3-o- ne
17-undecanoate, was found to be about twice that of
methyltestosterone. Moreover, and with respect to both the
bucyclate and undecanoate, it was found that this oral activity was
maximized when the actives were formulated with an oily carrier.
Unexpectedly high levels of activity were also discovered in
connection with the parenteral administration of the bucyclate and
undecanoate. In contrast to the oral formulation, activity was
maximized when the parenteral formulation comprised the actives in
an aqueous carrier.
[0044] As a general statement, it was found that the effective oral
dosage of
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxy-4-estren-3-one
bucyclate for any hormone replacement therapy which requires an
androgen, e.g., the treatment of hypogonadism, will be about
one-fourth the oral dosage of methyltestosterone required to
provide the same effect. For example, and in the case of
hypogonadism, 7.alpha.,11.beta.-dimethyl-17.beta.-hydroxy--
4-estren-3-one bucyclate may be orally administered in
therapeutically effective amounts. More specifically, the oral
dosage may range from about 1 mg/day to about 25 mg/day,
advantageously from about 2 mg/day to about 20 mg/day, and
preferably up to about 15 mg/day. Administration of the undecanoate
to effect this therapy may be undertaken within the foregoing
bucyclate therapeutic dosage ranges, but is preferably undertaken
at relatively greater levels relative to that of the bucyclate due
to the undecanoate's slightly lower oral activity. For example, the
undecanoate may be administered at from about 1 mg/day to about 75
mg/day, advantageously from about 2 mg/day to about 50 mg/day, and
preferably up to about 25 mg/day.
[0045] The oral dosage regimens described herein, set forth on the
basis of milligrams/day, includes any dosage regiment is able to
provide that dosage level to a patient per day. For example, an
extended release formulation of the bucyclate or undecanoate would
not need to be administered each day, yet would provide the
required daily dosage. However, administration of the therapeutic
dosage on a daily basis the preferred method of treatment.
[0046] The effective oral dosage of bucyclate for the treatment of
cancer, e.g., breast cancer in women can vary, but will range from
at least about 10 mg/day, advantageously at least about 25 mg/day,
and preferably at least about 50 mg/day. Administration of the
undecanoate to effect this therapy may, as before, be undertaken
within the foregoing bucyclate therapeutic dosage ranges, but is
preferably undertaken at relatively greater levels relative to that
of the bucyclate. For example, the undecanoate may be administered
in an amount of at least about 20 mg/day, advantageously at least
about 50 mg/day, and preferably at least about 100 mg/day.
[0047] In the use of
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxy-4-estren-- 3-one
bucyclate for male contraception, an effective oral dose may range
from about 1 mg/day to about 25 mg/day, advantageously from about 2
mg/day to about 20 mg/day, and up to about 15 mg/day.
Administration of the undecanoate to effect this therapy may, as
before, be undertaken within the foregoing bucyclate therapeutic
dosage ranges, but is preferably undertaken at relatively greater
levels relative to that of the bucyclate. For example, the
undecanoate may be administered in an amount ranging from about 1
mg/day to about 50 mg/day, advantageously from about 2 mg/day to
about 40 mg/day, and up to about 30 mg/day.
[0048] In the case of conditions requiring chronic hormonal
therapy, such as hypogonadism, an injectable bucyclate and/or
undecanoate formulation is preferably administered. This preference
is based upon the unexpected discovery that these actives are
surprisingly potent and long-acting when dispersed (preferably as a
suspension) in an aqueous formulation. Given these properties, the
bucylate and undecanoate may be administered in an aqueous
formulation at lower doses compared to both testosterone enanthate
(in an oily carrier) and testosterone bucyclate, and at relatively
long intervals. More specifically, and by way of comparative
example, doses of the bucyclate and undecanoate, when dispersed in
an aqueous formulation, may generally range from about one-third to
about three-quarters the dose of testosterone enanthate (provided
in a sesame oil carrier) required to provide substantially
equivalent therapeutic results, with between about one-half and
about two-thirds of that latter dose being preferred. With respect
to testosterone enanthate, the bucyclate and undecanoate, when
dispersed in an aqueous carrier, may be administered at between
about one-quarter and about one-half of the dose of testosterone
enanthate to provide substantially equivalent therapeutic effects.
However, if either active is formulated in a non-aqueous carrier,
e.g., an oily carrier comprised of sesame or other vegetable oils,
it was discovered that its potency over long periods remained, but
that it was substantially equivalent to that of testosterone
enanthate in a sesame oil carrier.
[0049] Because of its long-acting androgenic activity, particularly
when administered parenterally in an aqueous carrier in effective
amounts, the bucyclate and undecanoate may be administered at
intervals equal to, or in excess of, about two weeks. More
specifically, they may be administered at intervals of about one
month, preferably about two months, and most preferably once about
every three months. This provides a significant advantage to a
patient relative to existing regimens which require therapeutic
injections on a more frequent basis.
[0050] Again, the dosage of either the bucyclate or undecanoate
administered parenterally in an aqueous formulation at any
intervals will be significantly less than the amount of
testosterone enanthate used to achieve substantially similar
therapeutic results. For example, in treating hypogonadism, the
bucyclate and undecanoate may be administered in amounts ranging
from about 1 mg up to about 100 mg about every two weeks, and
advantageously from about 25 to about 75 mg during that period; up
to about 200 mg about every month, and advantageously from about 50
mg to about 150 mg during that time period; up to about 400 mg
about every 2 months, and advantageously from about 100 to about
300 mg during that time period; and up to about 600 mg about every
3 months, and advantageously from about 150 mg to about 450 mg
during that time period. These dosages, advantageously provided by
a single injection at the beginning of each time period, are less
than the dosages of testosterone enanthate and testosterone
bucyclate that may be used to provide similar therapeutic effects
over the same periods.
[0051] By way of further example, dosages of the bucyclate or
undecanoate effective for male contraception via parenteral
administration, if used alone, may range from about 25 mg/week up
to about 200 mg/week, advantageously up to about 150 mg/week, and
preferably from about 50 mg/week to about 100 mg/week. If used in a
more typical manner, i.e., combined with estrogen and/or
progestins, parenteral dosages of the bucyclate or undecanoate may
range from about 1 mg up to about 100 mg every about two weeks,
advantageously from about 2 mg up to about 75 mg, and preferably up
to about 50 mg, every two weeks. Of course, because of the
long-acting activity of the bucyclate and undecanoate, these
dosages may be administered on a substantially linear basis if
activity beyond the periods set forth above.
[0052] The enhanced potency of the bucyclate and undecanoate
advantageously permits a further advantage in that effective
amounts may be administered via a single injection, which is
desirable from a patient comfort and cost perspective. Equivalent
therapeutic results using testosterone enanthate would require
multiple injections. Of course, multiple injections of relatively
lower doses of the bucyclate or undecanoate may be administered if
required or desired.
[0053] While the bucyclate or undecanoate may be administered alone
in the treatment of cancer, it is preferably administered in
coordination with one or more anti-cancer agents, e.g.,
therapeutically-effective amounts of chemotherapeutic agents, such
as, cisplatin, carboplatin, doxorubicin, paclitaxel, taxotere,
methotrexate, fluorouracil, camptothecin, cyclophosphamide and
mixtures thereof, as well as therapeutically-effecti- ve amounts of
anti-angiogenesis agents, either alone or in combination. The
identity of suitable anti-tumor and anti-angiogenesis agents and
associated dosage regimens are well known, and as such will not be
repeated herein. The timing of administration of the foregoing
agents may occur at any time so long as the administration does not
interfere with the inventive therapeutic methods.
[0054] While the bucyclate and undecanoate may be prepared using
any suitable process, a further aspect of the present invention is
the discovery of the preferred synthesis routes described below,
which provide these actives in relatively high yield, and in solid
form, preferably in crystalline form, at room temperature. The
preparation of these actives in solid form at room temperature was
significant, as it led to the further discovery that the
long-acting effect of these actives are enhanced when included in
an injectable formulation at an average particle diameter of from
about 1-50 .mu.m, and preferably from about 3-30 .mu.m. The average
particle diameter of the bucyclate or undecanoate when formulated
as an injectable is thus preferably within the foregoing
ranges.
[0055] As a solid at room temperature, the bucyclate and
undecanoate stand in marked contrast to testosterone enanthate. The
latter exists as a liquid at room temperature, adversely affecting
its activity over long periods of time. Further, the enanthate is
precluded from commercialization as a lyophilizate or powder for
reconstitution, or as a tablet, caplet or other solid dosage
form.
[0056] Turning to FIG. 11, the preferred synthesis of
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxy-4-estren-3-one
bucyclate is depicted. Generally, this synthesis comprises the
steps of:
[0057] (a) converting the ether group of Compound 1 2
[0058] to a carbonyl group, providing Compound 2 3
[0059] (b) ketalizing the carbonyl group of Compound 2 to provide
Compound 4 4
[0060] (c) epoxidizing Compound 4 to provide the epoxide of
Compound 5 5
[0061] (d) opening the epoxide ring in Compound 5 and substituting
an alkyl group at C.sub.11 to provide Compound 6 (comprising a
mixture of 11.alpha.- and 11.beta.-methyl isomers, Compounds 6a and
6b, respectively) by use of a Grignard reagent 6
[0062] (e) deketalizing and dehydrating Compound 6 to provide
Compound 7 (comprising a mixture of 11.alpha.- and 11.beta.-methyl
isomers, Compounds 7a and 7b, respectively) 7
[0063] (f) converting Compound 7a to Compound 9 8
[0064] (g) converting Compound 9 to Compound 10 9
[0065] and (h) esterifying Compound 10 to provide Compound I
(7.alpha.,11.beta.-dimethyl-17.beta.-hydroxy-4-estren-3-one
bucyclate).
[0066] Step (a) may also yield an undesirable by-product, Compound
3 10
[0067] If desired, before step (f), one may ketalize the 11.alpha.-
and 11.beta.-methyl isomers of Compound 7 to provide Compound 8
11
[0068] and then deketalize and epimerize Compound 8, thereby
enhancing the ratio of the desirable 11.alpha.-methyl isomer to
11.beta.-methyl isomer.
[0069] Turning to FIG. 16, a preferred synthesis route for the
preparation of the undecanoate is set forth. This synthesis
comprises steps (a)-(g) used in the bucyclate synthesis as set
forth above. Thereafter, however, Compound 10 is esterified to
provide Compound II (the undecanoate).
[0070] One or more of the intermediates formed during the foregoing
synthesis routes are also contemplated as part of the present
invention, and particularly the preferred crystalline forms of
those intermediates. In addition, certain of the process steps, and
combinations thereof, which provide advantages such as relatively
high yields and/or purities of intermediates, constitute further
aspects of the present invention.
[0071] A pharmaceutically acceptable carrier is advantageously
combined with each active to ease the administration of the active
to a patient in need. Suitable carriers for oral and buccal dosage
forms, such as tablets, capsules, caplets and soft gelcaps (having
an oily carrier), are well known, and may be used in connection
with the actives. Preferably, oral dosage formulations of the
bucyclate and/or undecanoate include an oily carrier, and are
provided in the form of a soft gelcap, as this formulation was
found to enhance the beneficial properties of the actives upon oral
administration. Illustrative of oily substances that may be used to
provide an oily carrier include, but are not limited to, vegetable
oils, e.g. olive oil, safflower oil, corn oil, sunflower oil,
cotton seed oil, tsubaki oil, rice bran oil, soybean oil, sesame
oil, wheat germ oil, coconut oil, peanut oil, rape seed oil and the
like, fish oils, e.g., cuttlefish oil, cod oil, and the like, liver
oils, e.g., shark liver oil, cod liver oil and the like, blubber
oils, e.g., seal oil, blue whale oil, etc.), conchiferas oils,
e.g., abalone oil, oyster oil, and the like, medicinal oily
substances, e.g., castor oil, fatty acid glycerides, vitamin E,
vitamin A, vitamin K, and the like, polyethylene glycol and the
like, and mixtures thereof.
[0072] For parenteral administration, any type of carrier that
maintains the benefits of the invention as described herein may be
used. Preferably, however, and as previously mentioned, the
bucyclate and/or undecanoate is suspended in an aqueous carrier
suitable for injection. The water component of the aqueous carrier
should constitute at least half thereof, on a weight percent basis,
preferably at least about 80 wt. %, and more preferably at least
about 90 wt. % of the aqueous carrier. Illustrative of a preferred
parenteral formulation is one that includes up to 300 mg of the
active suspended in about 1 ml of an aqueous carrier. An
illustrative aqueous carrier may be prepared by combining: 1 g
benzyl alcohol, 0.5 g sodium carboxylethyl cellulose 50, 0.376 g
disodium hydrogen phosphate dihydrate, 1.495 g sodium dihydrogen
phosphate dihydrate, with water for injection (WFI) being added to
bring volume of the aqueous carrier up to 100 ml.
[0073] When formulated as an injectable, the active may be provided
in any suitable form, e.g., lyophilizate, dry powder for
reconstitution, a ready-to-use liquid, and in any suitable
container, e.g., vial, pre-filled syringe, or the like.
[0074] The actives may also be administered transdermally.
Transdermal delivery devices are well known. Illustrative
transdermal devices are described in U.S. Pat. Nos. 5,635,203 and
6,024,976. When a transdermal delivery device is used, the amount
of the bucyclate and/or undecanoate included in the device for
therapy should range from about 5% to about 25% of the parenteral
dose, and preferably from about 10% to about 20% of that dose, as
set forth herein.
[0075] The following examples are provided as further illustration
of the present invention, but should not be construed as limiting
the invention in any respect.
EXAMPLE 1
[0076] This example provides data on the androgenic potency of
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxy-4-estren-3-one
bucyclate(CDB-4386A), its free alcohol (CDB-1321D), testosterone
bucyclate (CDB-1781V-1), methyltestosterone (CDB-110), testosterone
(CDB-111 C) and testosterone enanthate (CDB-112a) when administered
orally.
[0077] Immature (about 21-day-old) Sprague-Dawley rats were
orchidectomized under anesthesia, and randomly assignee to groups
often animals for each dose level of the active undergoing testing.
Each active was dissolved in 10% ethanol/sesame oil and
administered by gavage (oral) each day for seven days beginning on
the date of the orchidectomy. The animals were sacrificed 24 hours
after the last dose, and the ventral prostate and seminal vesicles
were excised, cleaned of fat and connective tissue, blotted on
moist filter paper and weighed to the nearest 0.1 mg. See, e.g.,
Hershberger, L. et al, Myotrophic Activity of 19-nortestosterone
And Other Steroids Determined By Modified Levator And Muscle
Method, Proc. Soc. Exptl. Biol. Med. 83 175-180 (1953). Regression
analysis was performed by conventional methods using a PROPHET data
management system. See, e.g., Bliss, C., The Statistics of Bioassay
(Academic Press, New York, 1952); Hollister, C., Nucleic Acids Res.
16 1873-75 (1988). Ventral prostate weight was used as the endpoint
because it is the sensitive organ to androgenic stimulation.
[0078] The data obtained from this study is presented in graphic
form in FIGS. 3 and 4. This data indicates that the oral androgenic
activity of
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxy-4-estren-3-one
bucyclate is about 4 times (3.77 times, at a 95% confidence
interval 2.25-6.33) as potent as methyltestosterone and at least 4
times as potent as the free alcohol (1321D), and testosterone
bucyclate (1781V-1).
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxy-4-estren-3-one
bucyclate is also about 10-100 times more potent than testosterone
itself (111-C) or testosterone enanthate (112a) administered
orally.
EXAMPLE 2
[0079] This example provides data that demonstrates the duration of
activity of
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxy-4-estren-3-one
bucyclate (CDB-4386A) compared to its free alcohol (CDB-1321D), the
11.alpha.-methyl analog of
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxy-4-- estren-3-one
bucyclate (CDB-4386), testosterone bucyclate (CDB-1781a, -1781V2),
and testosterone enanthate (CDB-112E) when administered
parenterally (by subcutaneous injection).
[0080] Immature (about 21-day-old) Sprague-Dawley rats were
orchidectomized under anesthesia, and randomly assignee to groups
often animals for each dose level of the active undergoing testing.
Each active was administered by subcutaneous injection each day for
seven days beginning on the date of the orchidectomy. The animals
were sacrificed 24 hours after the last dose, and the ventral
prostate and seminal vesicles were excised, cleaned of fat and
connective tissue, blotted on moist filter paper and weighed to the
nearest 0.1 mg. Regression analysis was performed by conventional
methods using a PROPHET data management system. Ventral prostate
weight was used as the endpoint because it is the sensitive organ
to androgenic stimulation.
[0081] Except for testosterone enanthate, each active was
formulated in two different carriers: (1) an aqueous suspension and
(2) in sesame oil. Testosterone enanthate was formulated using the
sesame oil carrier only, because it exists as a liquid at room
temperature and could not therefore be formulated as an aqueous
suspension.
[0082] The carrier used to provide the aqueous suspension was
formulated as follows: 1 g benzyl alcohol, 0.5 g sodium
carboxylethyl cellulose 50, 0.376 g disodium hydrogen phosphate
dihydrate, 1.495 g sodium dihydrogen phosphate dihydrate, with
water for injection (WFI) being added to bring volume of the
carrier up to 100 ml.
[0083] Each formulation was prepared at a concentration of 0.6
mg/0.2 ml. To obtain further comparative data, testosterone
bucyclate was formulated in the aqueous suspension at a higher dose
(1.0 mg/0.2 ml).
[0084] The results, shown graphically in FIG. 5, substantiate the
unexpected activity of 7.alpha.,
11.beta.-dimethyl-17.beta.-hydroxy-4-est- ren-3-one bucyclate
(CDB-4386A) as compared to other androgenic esters. The former
exhibits activity in both potency and duration that far exceeds the
activity exhibited by the comparative esters when administered in
the same amounts, and particularly when
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxy-4-estren-3-one
bucyclate is formulated as an aqueous suspension. The activity of
even CDB-4386, which may be referred to as "close" to
7.alpha.,11.beta.-dimethyl-17.beta.-hydr- oxy-4-estren-3-one
bucyclate from a chemical structure perspective, nevertheless
exhibits relatively low activity as compared to
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxy-4-estren-3-one
bucyclate.
[0085] Further, both the potency and long-term activity of the
higher dosage of testosterone bucyclate (1.0 mg) was significantly
less than that provided by the lower dosage of
7.alpha.,11.beta.-dimethyl-1 7.beta.-hydroxy-4-estren-3-one
bucyclate (0.6 mg) in an aqueous suspension.
EXAMPLE 3
[0086] This example illustrates the relative androgenic activity of
testosterone and its derivatives.
[0087] Immature (about 21-day-old) Sprague-Dawley rats were
orchidectomized under anesthesia, and randomly assignee to groups
often animals for each dose level of the active undergoing testing.
Each active was dissolved in 10% ethanol/sesame oil and
administered by gavage (oral) or subcutaneous injection each day
for seven days beginning on the date of the orchidectomy. The
animals were sacrificed 24 hours after the last dose, and the
ventral prostate and seminal vesicles were excised, cleaned of fat
and connective tissue, blotted on moist filter paper and weighed to
the nearest 0.1 mg. Regression analysis was performed by
conventional methods using a PROPHET data management system.
Ventral prostate weight was used as the endpoint because it is the
sensitive organ to androgenic stimulation.
[0088] FIGS. 6 and 7 are graphic representations of the androgenic
assays of the actives. Each data point represents the mean (n=10)
and standard error of the mean (SEM) for each prostate weight for
each dose level.
[0089] From the data,
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxy-4-estren- -3-one
bucyclate (CDB-4386A) exhibited almost four times the oral activity
of methyltestosterone (CDB-110) (3.77 times, at 95% C.I. 2.3-6.3),
the current oral standard. However,
7.alpha.,11.beta.-dimethyl-17.beta.-hydro- xy-4-estren-3-one
bucyclate demonstrated only 0.4 times the activity of testosterone
(CDB-111C) following subcutaneous administration (0.4 times, at 95%
C.I. 0.2-0.6). The oral findings were unexpected because
testosterone and its esters exhibit low activity upon oral
administration.
[0090] The relatively weak activity upon subcutaneous
administration was also unexpected in view of the results on the
long-acting activity of
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxy-4-estren-3-one
bucyclate in Example 5. Testosterone, on the other hand, exhibited
the expected level of activity after subcutaneous injection. The
weak activity of the 11.alpha.-methyl analog (CDB-4415) of
7.alpha.,11.beta.-dimethyl-17.beta.- -hydroxy-4-estren-3-one
bucyclate (CDB-4386A) after subcutaneous administration indicates
the importance of the stereoconfiguration of the
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxy-4-estren-3-one
bucyclate (CDB-4386A) molecule.
[0091] Although not desiring to be bound to any particular theory,
the oral activity of
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxy-4-estren-3-o- ne
bucyclate may be due to its resistance to degradation in the
gastrointestinal tract and/or rapid metabolism by the liver. It is
also possible that the lipophilic nature of
7.alpha.,11.beta.-dimethyl-17.beta- .-hydroxy-4-estren-3-one
bucyclate permits absorption of the active into the thoracic lymph,
thereby avoiding direct entrance into the portal system and
metabolism by the liver.
[0092] Further, the lack of activity experienced by
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxy-4-estren-3-one
bucyclate under subcutaneous administration may reflect the slow
release, and possibly metabolism, of the active from the injection
site over the relatively brief 7-day administration period. This
same property, however, conveys long-acting activity on
7.alpha.,11.beta.-dimethyl-17.be- ta.-hydroxy-4-estren-3-one
bucyclate after parenteral administration in an aqueous
vehicle.
[0093] In addition to the foregoing, the androgenic potency and
relative binding affinity to the androgen receptor of several free
alcohols after subcutaneous administration of their corresponding
esters was also determined. The results are presented in the
following Table.
1 Activity of Ester Compound Corresponding Alcohol Compound Melting
Point Relative Binding Androgenic ID (.degree. C.) Affinity.sup.1
Potency.sup.2 A 68-69 91 8.1.sup.3 B 129-130 no data 1.2 C oil 148
61.1 D 108 no data 36.4-61.7.sup.3 E 99-100 1 no data F 130-132 82
19.3 G 134-136 28 1.0 (assigned) .sup.1From rat prostate; relative
to methyltrienolone = 100% (androgenic potency = 5.0) .sup.2Ventral
prostate weight assay following subcutaneous administration.
.sup.3Did not pass one or more significance tests (p < 0.05)
.sup.4Reference compound A:
7.alpha.-Methyl-19-nortestosterone-17.beta.-bucyclate B:
7.alpha.-Methyl-5.alpha.-dihydro-19-nortestosterone-17.beta.-bucyclate
C: 7.alpha.-Methyl-14-dehydro-testosterone-17.beta.-bucyclate D:
7.alpha.-Methyl-D-homo-testosterone-17.beta.-bucyclate E:
7.alpha.,11.alpha.-Dimethyl-19-nortestosterone-17.beta.-bucyclate
F: 7.alpha.,11.beta.-dimethyl-17.beta.-hydroxy-4-estren-3-one
bucyclate (CDB-4386A) G: Testosterone bucyclate
[0094] The foregoing data demonstrates that the activity of a
particular androgenic bucyclate ester (such as
7.alpha.,11.beta.-dimethyl-17.beta.-h- ydroxy-4-estren-3-one
bucyclate, CDB-4386A) cannot be predicted on the basis of the
androgenic activity of its corresponding free alcohol. More
specifically, the superior activity of
7.alpha.,11.beta.-dimethyl-17.beta- .-hydroxy-4-estren-3-one
bucyclate could not have been predicted from this data.
EXAMPLE 4
[0095] This example further illustrates the relative activity of
various testosterone esters, including 7.alpha.,
11.beta.-dimethyl-17.beta.-hydro- xy-4-estren-3-one bucyclate, over
relatively long periods of time.
[0096] Immature (about 21-day-old) Sprague-Dawley rats were
orchidectomized under anesthesia, and randomly assignee to groups
of 40 or more. Animals received a single subcutaneous injection of
0.6 mg of each ester in 0.2 ml of an aqueous suspending carrier
and/or oily carrier (10% ethanol/90% sesame oil or ethyl oleate) on
the date of the orchidectomy. In cases where the ester was not
solid at room temperature, 10% ethanol/sesame oil or ethyl oleate
was used as the carrier. In this example, the carrier used to
provide the aqueous suspension was formulated as follows: 1 g
benzyl alcohol, 0.5 g sodium carboxylethyl cellulose 50, 0.376 g
disodium hydrogen phosphate dihydrate, 1.495 g sodium dihydrogen
phosphate dihydrate, with water for injection (WFI) being added to
bring volume of the carrier up to 100 ml.
[0097] Five animals from each group were sacrificed at weekly or
biweekly intervals, and the ventral prostate and seminal vesicles
were excised, cleaned of fat and connective tissue, blotted on
moist filter paper and weighed to the nearest 0.1 mg.
[0098] Ventral prostate weight was used as the endpoint because it
is the sensitive organ to androgenic stimulation. Regression
analysis was performed by conventional methods using the PROPHET
data management system previously identified.
[0099] FIGS. 8 and 9 are graphic representations of the androgenic
assays of the actives. Each data point represents the mean (n=10)
and standard error of the mean (SEM) for each prostate weight for
each dose level.
[0100] FIG. 8 is a graph of the ventral prostate weights at weekly
intervals over a 10 week period after the subcutaneous
administration of
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxy-4-estren-3-one
bucyclate (CDB4386A) in both oily and aqueous carriers, its
11.alpha.-methyl analog (CDB-4386) in both carriers, and
testosterone enanthate (CDB-112E) in an oily carrier.
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxy-4-estren-3-one
bucyclate in the aqueous vehicle exhibited the most dramatic
increase and maintenance of ventral prostate weight. The area under
the curve (AUC, calculated by the trapezoidal rule), was about 3
times greater for
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxy-4-estren-3-one
bucyclate than for testosterone enanthate in sesame oil. The
11.alpha.-methyl analog was inactive in this experiment, with
evaluation being discontinued 8 weeks after administration. This
experiment highlights the significance of the ability to provide
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxy-4-estren-3- -one
bucyclate in the form of an aqueous suspension, which provides
unexpected and desirable long-term androgenic activity. This
experiment also underscores the importance of the
stereoconfiguration of the C.sub.11 substituent.
[0101] FIG. 9 is a graph of the ventral prostate weights at various
time intervals up to 20 weeks after administration of several
different bucyclate esters: 7
.alpha.,11.beta.-dimethyl-17.beta.-hydroxy-4-estren-3- -one
bucyclate (CDB-4386A),
7.alpha.-Methyl-14-dehydro-19-nortestosterone-- 17.beta.-bucyclate
(CDB-4327A), 7.alpha.-Methyl-19-nortestosterone-170-buc- yclate
(CDB-4288),
7.alpha.-Methyl-16-dehydro-D-homo-19-nortestosterone-17-
.beta.-bucyclate (CDB-4318) and 7.alpha.-Methyl-5
.alpha.-dihydro-19-norte- stosterone-170-bucyclate (CDB-4289). All
esters other than
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxy-4-estren-3-one
bucyclate were administered in the oily carrier because they do not
exist as solids at room temperature, or possess low melting points.
7.alpha.,11.beta.-dimeth- yl-17.beta.-hydroxy-4-estren-3-one
bucyclate, suspended in the aqueous carrier, exhibited the greatest
AUC over the 10-week period for which this parameter was
calculated. CDB-4327A demonstrated surprising stimulation of
ventral prostate size over the entire 20-week observation period,
however, this is one of the most active synthetic androgens
presently known. The remaining actives showed relatively weak
activity. This experiment can be said to demonstrate that the
prediction of activity cannot be based on the structure of the
active, or on the carrier used in connection with the
administration of the active.
[0102] Serum samples taken from the animals at autopsy showed the
presence of the free alcohol
(7.alpha.,11.beta.-dimethyl-19-nortestosterone) which decreased
with time over the 10 week observation period. The results are
provided in FIG. 10.
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxy-4-estren- -3-one
bucyclate, suspended in the aqueous carrier, provided the highest
levels of the free alcohol, and maintained these relatively high
levels over the 10-week observation period.
EXAMPLE 6
[0103] This example describes a preferred process for synthesizing
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxy-4-estren-3-one
bucyclate (Compound I). Reference may be made to FIG. 11.
A. Preparation of 7.alpha.-Methylestra-4,9-diene-3,17-dione
(Compound 2)
[0104] Under a nitrogen flush through an inverted plastic funnel
the antimony pentafluoride (110 mL, 5.79 mol) was weighed into a
Teflon jar. Hydrogen fluoride (436 nL, 21.8 mol), chilled to
4.degree. C., was first collected in a Teflon separatory funnel,
then added with extreme care to the reaction vessel under a
nitrogen flush. Failure to assure rapid mixing can result in an
eruption. As the mixture was stirred, it was cooled to 0.degree. C.
for 20 min. 7.alpha.-methylestrone methyl ether (Compound 1, 25.0
g, 83.8 mmol) was carefully added under nitrogen. The reaction was
stirred at 0.degree. C. for 2.5 hr, after which it was slowly
poured into a plastic beaker containing a mixture of saturated
potassium carbonate (300 mL, 900 g/1000 mL) and ice. Additional
potassium carbonate was used to adjust the pH to ca. 8. This
mixture was then extracted with methylene chloride (3.times.) and
the organic portions were washed with water and brine. After drying
over sodium sulfate, the solvent was removed in vacuo to give 24.8
g of crude oil. This crude material contains Compound 2 and an
isomeric by-product, 4,6-diene-dione-3,20 (Compound 3) in a 2:1
ratio. Therefore, the crude material was subjected to dry column
chromatography on silica gel (63-206 mesh) eluted with 3% acetone
in CH.sub.2Cl.sub.2. This gave a segment which contained 15 g of
the desired product (Compound 2). After extraction, evaporation of
the solvent followed by trituration with ether afforded 9.24 g of
Compound 2 in 38.8% yield. The mother liquor from this material was
combined with the other principle portion of the column, and was
rechromatographed using the same conditions. Trituration of the
segments provided an additional 0.19 g of the desired product
(Compound 2). Total amount was 9.43 g in 39.6% yield; m.p.
204-205.degree. C. (Lit. m.p.=(8). FRIR (KBr, diffuse reflectance):
.nu.max, 3454, 3282, 3030, 2968, 2928, 2902, 1737, 1652, 1600, and
1580 cm.sup.-1. NUR (.sup.1H, CDCl.sub.3) .delta. 0.859 (d, 3H,
J=3.5 Hz), C7.alpha.-CH.sub.3), 1.001 (s, 3H, C18-CH.sub.3) and
5.726 (s, 1H, C4-CH). NMR (.sup.13C, CDCl.sub.3) .delta. 12.641,
21.835, 24.885, 25.576, 28.021, 30.626, 35, 621, 36.893, 39.416,
42.4779, 45.966, 123.259 (C-4), 126.081 (C-10).sub.m 140.295 (C-9),
154.572 (C-5), 199.052 (C-3) and 219.633 (C-17).
B. The Preparation of
3,3-Ethylenedioxy-7.alpha.-methyl-17.beta.-hydroxyes-
tra-5(10),9(11)-diene (Compound 4)
[0105] A THF (500 mL) solution of the dione (Compound 2, 10.0 g,
35.16 mmol) was chilled to 0.degree. C. and treated dropwise with a
TBF solution of lithium tri-tert-butoxyaluminum hydride (1.0 M/THF,
40.0 mL. 8.9 mmol). The mixture was stirred at 0.degree. C. for 2
hr. EtOAc (10.0 mL) was added and most of the solvent was removed
in vacuo. The residue was diluted with cold 0.1 N HCl and the
aqueous mixture was extracted with EtOAc (3.times.). The EtOAc
layers were washed with water and brine, combined, and dried over
sodium sulfate. Evaporation of the solvent gave 10.61 g of a stable
foam. The material was then dissolved in benzene (1 L). Ethylene
glycol (10.0 mL) was added, followed by p-toluenesulfonic acid (500
mg). The resulting mixture was heated to reflux while draining off
approximately 500 mL of benzene from the Dean-Stark trap. The
mixture was cooled and diluted with saturated sodium bicarbonate
solution. The benzene solution was washed with water and brine. The
aqueous washes were extracted with EtOAc (2.times.). The combined
organic extracts were dried over sodium sulfate. Evaporation of the
solvent gave the 12.61 g of a stable foam. The material was
chromatographed (5% acetone in CH.sub.2Cl.sub.2) to afford 10.05 g
of the ketal (Compound 4) in 87% yield. NMR (CDCl.sub.3) .delta.
0.725 (s,3H, C18-CH.sub.3), 0.727 (d,3H, J=7.2 Hz,
C7.alpha.-CH.sub.3), 3.777(t,1H, J=8.7 Hz, C17.alpha.-CH), 3.979
(m, 4 H, 3-ketal) and 5.638 (m, 1H, C11=CH). MS (EI) m/z: relative
intensity: 330 (M.sup.+).
C. Preparation of
3,3-ethylenedioxy-7.alpha.-methyl-5.alpha.,10.alpha.-epo-
xy-17.beta.-hydroxyestra-9(11)-ene (Compound 5)
[0106] A solution of hexafluoroacetone (30.0 g, 136.2 mmol) in
CH.sub.2Cl.sub.2, (150 m.L) was chilled to 0.degree. C. With
vigorous stirring, 30% hydrogen peroxide (14.0 mL, 136.2 mmol) and
solid disodium hydrogen phosphate (5.86 g, 41.30 mmol) was added.
The resulting mixture was stirred at 0.degree. C. for 1/2 hr. A
solution of the ketal (Compound 4, 15.0 g 45.39 mmol) in
CH.sub.2Cl.sub.2 (300 mL) was added and the mixture was stirred at
4.degree. C. for 24 hr. The mixture was then diluted with 10%
sodium sulfite solution, and subsequently extracted with
CH.sub.2Cl.sub.2 (3.times.). The extracts were washed with water
and brine, combined and dried over sodium sulfate. Evaporation of
the solvent gave 16.26 g. of Compound 5. This material was used
without further purification in the subsequent reaction. NMR
(CDCl.sub.3) .delta. 0.725 (s, 3H, C18-CH.sub.3), 0.762 (d, 3H,
J=7.2 Hz, C7.alpha.-CH.sub.3), 3.758 (t, 1H, J=8.7 Hz,
C17.alpha.-CH), 3.895 (d, 4H, 3-ketal) and 6.00 (m, 1H,
C11=CH).
D. Preparation of
3,3-ethylenedioxy-7.alpha.,11.beta.-dimethyl-5.alpha.,17-
.beta.-dihyrdoxyestra-9-ene (Compound 6)
[0107] A solution of methylmagnesiumbromide (1.4 M THF/toluene, 210
mL, 295 mmol) was added to THF (150 mL) and copper (I) chloride
(2.92 g, 29.5 mmol) was added. After stirring at room temperature
for 1/2 hr, a solution of the epoxide (Compound 5, 16.26 g 46.99
mmol) in THF (450 mL) was added dropwise over 5 min. The mixture
was stirred at room temperature for 3 hr. The mixture was diluted
with saturated ammonium chloride solution and air was bubbled
through the mixture for 12 hr to oxidize Cu(I) to Cu(II). The
aqueous mixture was extracted with ether (3.times.). The ether
extracts were washed with water and brine, combined, and dried over
sodium sulfate. Evaporation of the solvent gave 16.70 g of a yellow
semi-solid. The material was triturated with ether and the solid
was filtered to afford 8.86 g of a mixture of Grignard products
(7.alpha.,11.alpha.-dimethyl and 7.alpha.,11.beta.-dimethyl,
referred to as Compounds 6a and 6b, respectively). Evaporation of
the filtrate gave 7.4 g of a stable foam. Total amount was 16.26 g
in quantitative yield.
E. Hydrolysis of a Mixture of Compounds 6a and 6b to Isomeric
Compounds 7b and 7a (as a ca. 3/7 Mixture)
[0108] The solid (containing Compounds 6a and 6b) from Step D above
(16.26 g, 54.2 mmol) was dissolved in acetic acid/THF/water (3:1:1,
500 mL) and heated to reflux for 2 hr. The solvent was evaporated
in vacuo and the mixture was diluted with saturated sodium
bicarbonate solution. The mixture was then extracted with
CH.sub.2Cl.sub.2. The CH.sub.2Cl.sub.2 extracts were washed with
water, brine, combined, and dried over sodium sulfate. Evaporation
of the solvent gave 7.45 g. The material was chromatographed (10%
acetone/methylene chloride) to afford 5.12 g of Compounds 7a and 7b
(7.alpha.,11.alpha.-dimethyl and 7.alpha.,11.beta.-dimethyl,
respectively). The foam obtained in Step D was treated in the same
manner to afford an additional 2.73 g of Compounds 7a and 7b after
chromatography. Total amount was 7.85 g in 45.9% yield. NMR
(CDCl.sub.3) .delta. 0.747 (d, 3H, J=7 Hz, C7.alpha.-CH.sub.3),
0.780 (s, 3H, C18-CH.sub.3 of Compound 6b), 0.963 (s, 3H,
C18-CH.sub.3 of Compound 6a), 1.077 (d, 3H, J=7 Hz,
C11.alpha.-CH.sub.3), 1.173 (d, 3H, J=7 Hz, C11.beta.-CH.sub.3),
and 3.770 (t, 1H, J=8.7 Hz, C17.alpha.-CH.
F. Preparation of
3,3-Ethyleniedioxy-7.alpha.,11-dimethyl-17.beta.-hydroxy-
estra-5(10),9(11)-diene (Compound 8)
[0109] A solution of Compounds 7a/7b (2.0 g 6.65 mmol) in benzene
(500 mL) was treated with ethylene glycol (5.0 mL) and
p-toluenesulfonic acid (250 mg). The mixture was heated at reflux
with azeotropic removal of water. Approximately 250 mL of solvent
was distilled off. The mixture was cooled to room temperature and
diluted with saturated sodium bicarbonate solution. The mixture was
extracted with EtOAc. The EtOAc extracts were washed with water and
brine, combined and dried over sodium sulfate. Evaporation of the
solvent gave 2.15 g of stable foam in 93.9% yield. The material was
homogeneous by TLC and less polar than the starting material. NNM
(CDCl.sub.3) .delta. 0.716 (s, 3H, C18-CH.sub.3), 0.725 (d, 3H.
J=7.2 Hz, C7.alpha.-CH.sub.3), 1.801 (br s, 3H, C11-CH.sub.3),
3.755 (t, 1H, J=8.7, C17.alpha.-CH) and 4.003 (m, 4H, 3-ketal).
G. Preparation of
7.alpha.,11.beta.-Dimethyl-17.beta.-hydroxyestra-4,9-die- ne-3-one
(Compounds 7b/7a ca. 10/1) via Hydrolysis of Compound 8
[0110] The ketal (Compound 8, 2.15 g 6.24 mmol) was dissolved in
methanol (200 mL) and 10.0 mL of 10% HCl was added. The solution
was heated at reflux for 18 hr. The solvent was evaporated in vacuo
and the residue was diluted with saturated sodium bicarbonate
solution. The aqueous mixture was extracted with CH.sub.2Cl.sub.2.
The methylene chloride extracts were washed with water and brine,
combined and dried over sodium sulfate. Evaporation of the solvent
gave 1.89 g of a stable foam. The material was chromatographed (10%
acetone in CH.sub.2Cl.sub.2) to afford 950 mg, of the
7.alpha.,11.beta.-dimethyl compound (Compound 7b) in 50.8% yield.
Also isolated 703 mg of a Compound 7a/7b mixture in which was
resubjected to the ketalization and equilibrium process to yield
additional material. Compound 7b: NMR (CDCl.sub.3) .delta. 0.790
(d, 3H, J=7.2 Hz, C7.alpha.-CH.sub.3), 0.963 (s, 3H, C18-CH.sub.3),
1.172(d, 3H, C11.alpha.-CH.sub.3), 3.186 (m, 5-lines, 1H,
C11.alpha.-CH), 3.661 (t, 1H, J=8.7 Hz, C17.alpha.-CH) and 5.702
(s, 1H, C4-CH).
H. Preparation of
7.alpha.,11.beta.-Dimethyl-17.beta.-hydroxy-4-estren-3-o- ne
(Compound 10)
[0111] Lithium wire (253 mg, 36.45 mmol), cut into small pieces,
was added to redistilled (from sodium) ammonia (300 mL) and the
mixture was stirred at ammonia reflux (-35.degree. C.) for V.sub.2
hr. The mixture was chilled to -78.degree. C. and a solution of the
dienone (Compound 7b, 3.65 g 12.15 mmol) in THF (300 mL) and
t-butanol (1.16 mL, 12.15 mmol) was added dropwise. Upon completion
of the addition, the reaction was stirred for 15 min before any
excess lithium was destroyed with the addition of isoprene (ca. 1.0
mL) and finally, quenched with the addition of solid ammonium
chloride (15 g). The ammonia was evaporated under argon gas and the
mixture was diluted with 0.1 N phosphate buffer, pH=7.0. The
mixture was extracted with ether. The ether extracts were washed
with water and brine, combined, and dried over sodium sulfate.
Evaporation of the solvent gave 3.83 g of Compound 9 as a light
yellow solid in quantitative yield. The material was homogeneous by
TLC and was used without further purification in the following
reaction. NMR (CDCl.sub.3) .delta. 0.812 (d, 3H, J=7.2 Hz,
C7.alpha.-CH.sub.3).sub.m 0.877 (s, 3H, C18-CH.sub.3), 0.903 (d,
3H, J=7.2 Hz, C11-CH.sub.3), 2.754 (br q, 2H, C4-CH.sub.2--), and
3.660 (t, 1H, J=8.8 Hz, C17.alpha.-CH).
[0112] The material prepared above was dissolved in methanol (400
mL) and 10% HCl (20 mL) was added and the mixture was heated at
reflux for 3 hr. The solvent was evaporated and the residue was
diluted with saturated sodium bicarbonate solution. The mixture was
extracted with CH.sub.2Cl.sub.2. The methylene chloride extracts
were washed with water and brine, combined and dried over sodium
sulfate. Evaporation of the solvent gave 3.81 g of a stable foam.
The material was chromatographed (10% acetone in CH.sub.2Cl.sub.2)
to afford 3.54 g of Compound 10 in 96.5% yield. The material was
recrystallized from ether/hexane to give 3.14 g of Compound 10 as
fine white needles in 86% yield; m.p.=155-157.degree. C. Analysis
by reverse phase HPLC on a NovaPak C.sub.18 column eluted with 50%
aqueous CH.sub.3CN at a flow rate of 1 mL per min and at
.lambda.=240 nm indicated this material to have a purity in excess
of 99% FTIR (KBr, diffuse reflectance): .nu..sub.max 3470, 2950,
1663 and 1622 cm.sup.-1. NMR (CDCl.sub.3) .delta. 0.770 (d, 3H,
J=7.2 Hz, C7.alpha.-CH.sub.3), 0.886 (s,3H, C18-CH.sub.3), 1.075
(d, 3H, J=7.2 Hz, C11.beta.-CH.sub.3), 3.626 (t, 1H, J=8.7 Hz,
C17.alpha.-CH) and 5.849 (br s, 1H, C4-CH). MS (EI) m/z relative
intensity: 302 (M.sup.+). Analysis calculated for
C.sub.20H.sub.30O.sub.2: C, 79.42; H, 10.00. Found: C, 79.18; H,
10.00.
I. Preparation of
7.alpha.,11.beta.-Dimethyl-17.beta.-hydroxy-4-estren-3-o- ne
17.beta.-trans-4-n-butylcyclohexane Carboxylate (Compound I)
[0113] Trans-4-n-Butylcyclohexanecarboxylic acid chloride (Compound
11, 2.25 g 110 mmol), dissolved in benzene (10 mL), was added to a
solution of
7.alpha.,11.beta.-Dimethyl-17.beta.-hydroxy-4-estren-3-one
(Compound 10, 608 mg, 2 mmol) in a mixture of benzene (100 mL) and
pyridine (5.0 mL). The mixture was stirred overnight at room
temperature. The mixture was chilled in an ice bath and diluted
with 1.0 N sodium hydroxide solution. The aqueous mixture was
extracted with ether. The ether extracts were washed with 1.0 N
sodium hydroxide solution (2.times.), water and brine. The combined
organic extracts were dried over sodium sulfate and evaporation of
the solvent gave 1.72 g of a semi-solid. Recrystallization of the
material (Compound I) from hexanes gave 765 mg of white powder in
82% yield: m.p.=130-132.degree. C. Analysis by reverse Phase HPLC
on a NovaPak C.sub.18 column eluted with CH.sub.3CN at a flow rate
of 1.25 mL per minute and at .lambda.=240 nm showed the Compound I
to be pure greater than 99%. FTIR (KBr, diffuse reflectance)
.nu..sub.max 2933, 1726, 1669 and 1621 cm.sup.1. MR (CDCl.sub.3)
.delta. 0.779 (d, 3H, J=7.2 Hz, C7.alpha.-CH.sub.3), 0.886 (t, 3H,
n-butyl CH.sub.3), 0.923 (s, 3H, C18-CH.sub.3), 1.057 (d, 3H, J=7.2
Hz, C11-CH.sub.3), 4.545 (t, 1H, J=8.7 Hz, C17.alpha.-CH) and 5.848
(br s, 1H, C4-CH). MS (EI) m/z relative intensity: 468 (M.sup.+,
6.9), 358 (65.3), 302 (12.5), 284 (20.8), 269 (6.9), 259 (12.5),
174 (62.5), 159 (26.5), 147 (19.4), 139 (25.0), 119 (18.1), 110
(75.0), 105 (8.1), 97 (36.1), 83 (100), 69 (38.9) and 55
(58.3).
EXAMPLE 7
[0114] This example provides data on the androgenic potency of the
undecanoate (CDB-4521A) relative to methyltestosterone (CDB-111C)
when administered via subcutaneous injection.
[0115] Immature (about 21-day-old) Sprague-Dawley rats were
orchidectomized under anesthesia, and randomly assignee to groups
of ten animals for each dose level of the active undergoing
testing. Each active was dissolved in 10% ethanol/sesame oil and
administered by subcutaneous injection each day for seven days
beginning on the date of the orchidectomy. The animals were
sacrificed 24 hours after the last dose, and the ventral prostate
and seminal vesicles were excised, cleaned of fat and connective
tissue, blotted on moist filter paper and weighed to the nearest
0.1 mg. See, e.g., Hershberger, L. et al, Myotrophic Activity of
19-nortestosterone And Other Steroids Determined By Modified
Levator And Muscle Method, Proc. Soc. Exptl. Biol. Med. 83 175-180
(1953). Regression analysis was performed by conventional methods
using a PROPHET data management system. See, e.g., Bliss, C., The
Statistics of Bioassay (Academic Press, New York, 1952); Hollister,
C., Nucleic Acids Res. 16 1873-75 (1988). Ventral prostate weight
was used as the endpoint because it is the sensitive organ to
androgenic stimulation.
[0116] The data obtained from this study is presented in graphic
form in FIG. 13. This data indicates that the subcutaneous
androgenic activity of the undecanoate (CDB-4521A) is about half
that of testosterone (0.52 times, at a 95% confidence interval,
0.29-0.93) when administered in the oily carrier. This data was
surprising when compared to the results obtained when the
undecanoate was administered in an aqueous carrier.
EXAMPLE 8
[0117] This example provides data on the androgenic potency of the
undecanoate (CDB-4521) and methyltestosterone (CDB-110) when orally
administered in oily or aqueous carriers.
[0118] Immature (about 21-day-old) Sprague-Dawley rats were
orchidectomized under anesthesia, and randomly assignee to groups
of ten animals for each dose level of the active undergoing
testing. Four dosage forms were prepared. The first two forms
constituted a solution of each active in 10% ethanol/sesame oil.
The third and fourth dosage forms constituted a suspension of each
active in an aqueous carrier (as described in Example 2, supra).
These dosage forms were then administered by gavage (oral) to
separate animal groups each day for seven days beginning on the
date of the orchidectomy. Each carrier was also administered
(alone) to separate groups of animals as a control. The animals
were sacrificed 24 hours after the last dose, and the ventral
prostate and seminal vesicles were excised, cleaned of fat and
connective tissue, blotted on moist filter paper and weighed to the
nearest 0.1 mg. See, e.g., Hershberger, L. et al, Myotrophic
Activity of 19-nortestosterone And Other Steroids Determined By
Modified Levator And Muscle Method, Proc. Soc. Exptl. Biol. Med. 83
175-180 (1953). Regression analysis was performed by conventional
methods using a PROPHET data management system. See, e.g., Bliss,
C., The Statistics of Bioassay (Academic Press, New York, 1952);
Hollister, C., Nucleic Acids Res. 16 1873-75 (1988). Ventral
prostate weight was used as the endpoint because it is the
sensitive organ to androgenic stimulation.
[0119] The data obtained from this study is presented in graphic
form in FIGS. 14 and 4. This data indicates that the oral
androgenic activity of the undecanoate (CDB-4521A) in the oily
carrier is about 2 times (2.36 times, at a 95% confidence interval)
as potent as methyltestosterone in the same oily carrier. In
contrast, oral administration of the undecanoate in the aqueous
carrier described in Example 2, supra, revealed a potency about the
same (0.95 times, at a 95% confidence interval, 0.36-2.5) as that
of methyltestosterone in the same aqueous carrier.
EXAMPLE 9
[0120] This example further illustrates the relative activity of
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxyestr-4-en-3-one
17-undecanoate (Compound II) compared to that of testosterone
enanthate (CDB-112F) over relatively long periods of time.
[0121] Immature (about 21-day-old) Sprague-Dawley rats were
orchidectomized under anesthesia, and randomly assignee to groups
of 40 or more. Animals received a single subcutaneous injection of
0.6 mg of the undecanoate in 0.2 ml of an aqueous suspending
carrier and/or oily carrier (10% ethanol/90% sesame oil containing
5 mg/ml chlorobutanol as a preservative, or ethyloleate) on the
date of the orchidectomy. The enanthate ester was formulated using
the 10% ethanol/sesame oil or ethyloleate carrier as a first
standard, with the 10% ethanol/sesame oil carrier used as a second
standard.
[0122] In this example, the carrier used to provide the aqueous
suspension was formulated as follows: 1 g benzyl alcohol, 0.5 g
sodium carboxylethyl cellulose 50, 0.376 g disodium hydrogen
phosphate dihydrate, 1.495 g sodium dihydrogen phosphate dihydrate,
with water for injection (WFI) being added to bring volume of the
carrier up to 100 ml.
[0123] Five animals from each group were sacrificed at weekly or
biweekly intervals, and the ventral prostate and seminal vesicles
were excised, cleaned of fat and connective tissue, blotted on
moist filter paper and weighed to the nearest 0.1 mg.
[0124] Ventral prostate weight was used as the endpoint because it
is the sensitive organ to androgenic stimulation. Regression
analysis was performed by conventional methods using the PROPHET
data management system previously identified.
[0125] FIG. 15 is a graphic representation of the androgenic assays
of the actives. Each data point represents the mean (n=10) and
standard error of the mean (SEM) for each prostate weight for each
formulation level.
[0126] More specifically, FIG. 15 is a graph of the ventral
prostate weights at weekly intervals over a 10 week period after
the subcutaneous administration of the undecanoate (CDB-4521) in
both oily and aqueous carriers, testosterone enanthate (CDB-112F)
in an oily carrier, and an oily carrier (10% ethanol/sesame oil)
alone. The undecanoate in the aqueous vehicle exhibited the most
dramatic increase and maintenance of ventral prostate weight. The
area under the curve (AUC, calculated by the trapezoidal rule), was
about 3 times greater for the undecanoate (1817 mg-weeks) than for
testosterone enanthate in the oily carrier (AUC 559 mg-weeks).
[0127] This experiment highlights the significance of the ability
to provide the undecanoate in the form of an aqueous suspension,
which provides unexpected and desirable long-term androgenic
activity. This experiment also underscores the importance of the
stereoconfiguration of the C.sub.11 substituent.
EXAMPLE 10
[0128] This example describes a preferred process for synthesizing
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxyestr-4-en-3-one
17-undecanoate (Compound II). Reference may be made to FIG. 16.
[0129] The synthesis of Compound 10 as described in Example 6 was
completed. Thereafter, the undecanoate was prepared by treatment of
Compound 10 with undecanoyl chloride in pyridine to provide
Compound II as a white powder, in good yield.
[0130] A solution of
7.alpha.,11.beta.-dimethyl-17.beta.-hydroxyestr-4-en-- 3-one
(Compound 10, 252 mg, 0.83 mmol) in a mixture of benzene (20 mL)
and pyridine (2.0 mL) was treated with undecanoyl chloride
(Compound 12, 500 mg, 2.44 mmol). The mixture was stirred at room
temperature overnight. The mixture was then chilled in an ice bath
and diluted with cold 0.1 N sodium hydroxide solution. The
resulting aqueous mixture was extracted with ether. The ether
extracts were washed with water and brine, combined and dried over
sodium sulfate. Evaporation of the solvent gave 525 mg of an oil.
The material was chromatographed using 10% acetone/CH.sub.2Cl.sub.2
to yield 398 mg of an oil. The material was recrystallized from
cold pentane to afford 369.2 mg of Compound II as a white powder in
94% yield; m.p.=62-64.degree. C. Analysis by reverse Phase HPLC on
a NovaPak C.sub.18 Column eluted with CH.sub.3CN at a flow rate of
1.0 mL per mm and at .lambda.=240 nm showed Compound II to have a
purity of at least 99.9%. FTIR (KBr, diffuse reflectance):
.nu..sub.max 2914, 1733, 1678 and 1628 cm.sup.-1. .sup.1HNMR
(CDCl.sub.3) .delta. 0.782 (d, 3H, J=7.2 Hz, C7.alpha.-CH.sub.3),
0.880 (t, 3H, J=9 Hz, --(CH.sub.2).sub.9CH3,), 0.922 (s, 3H,
C18-CH.sub.3), 1.058 (d, 3H, J=7.2 Hz, C 11.beta.-CH.sub.3), 4.565
(t, 1H, J=8.4 Hz, C17.alpha.-CH) and 5.849 (s, 1H, C4-CH.dbd.). MS
(EI) m/z (relative intensity): 470 (M.sup.+, 100), 302 (60), 284
(78), 259 (67), 175 (89), 110 (69) and 55 (96). Analysis Calculated
for C.sub.31H.sub.50O.sub.3: C, 79.10; H, 10.70. Found: C, 79.33;
H, 10.91.
[0131] Any reference cited herein, including patents, patent
applications, and publications, are hereby incorporated in their
entireties by reference. Further, any reference herein to a
component in the singular is intended to indicate and include at
least one of that particular component, i.e., one or more.
[0132] While this invention has been described with an emphasis
upon preferred embodiments, it will be obvious to those of ordinary
skill in the art that variations of the preferred embodiments may
be used and that it is intended that the invention may be practiced
otherwise than as specifically described herein. Accordingly, this
invention includes all modifications encompassed within the spirit
and scope of the invention as defined by the following claims.
* * * * *