Pharmaceutical compositions containing dehydroepiandrosterone and their delivery systems

Medical Applications and Delivery Systems of Dehydroepiandrosterone U ·

FIELD OF THE INVENTION The present invention relates to a method for preventing and / or treating tooth eruption, prevention or treatment of vaginal atrophy, hypogonadism (reduced function of sexual glands), decreased libido (sex drive), osteoporosis, urinary incontinence (insufficiency), ovarian cancer, , warm-blooded animals, including humans. This process comprises the addition of dehydroepiandrosterone (DHEA), dehydroepiandrosterone sulfate (abbreviated as DHEA-S), or substances that are converted to these compounds in vivo; the invention further encompasses pharmaceutical products, such as accessories, and pharmaceutical compositions useful for the delivery (delivery) of the active ingredient (s) for use in the present invention.

Primates are unique in that they have adrenal glands that secrete large amounts of steroid dehydroepiandrosterone (DHEA), and in particular DHEA sulfate (DHEA-S), which are androstenedione (abbreviated as D4-dione) or androstenediol D5-diol) and are then converted into efficient androgens and estrogens in peripheral tissues [Adams: Mole. Cell. Endocrinol 41. 1-17 (1985); and Labrie et al., Important Advances on Oncology '* (Oncology juice81671-5906-SZŐ-tree

2), S. Vita et al., JB. Lippincott, Philadelphia PA, 193-200. p. (1985)]. DHEA-S, which is a predominantly steroid in both men's and women's blood, is converted to DHEA and D5-diol in the peripheral tissues and thus maintains a close correlation between the levels of these three steroids in the blood [Adams: Mol. Cell. Endocrinol. 41, 1-17 (1985)]. Depending on the relative activity of 17β-hydroxy steroid dehydrogenase (17β-HSD), aromatase and 5α-reductase, DHEA and its derivatives are preferably converted to androgens and / or estrogens.

At birth, low serum concentrations of DHEA and DHEA-S persist until six years of age. Serum levels of these two steroids generally increase by the age of seven and continue to increase in both boys and girls until the age of 16 [Orentreich et al., J. Clin. Metab. Metab. 59: 551-555 (1984)]. In males, further growth is observed, peaking generally between the ages of 20 and 24. In women, after 16 years of age, no further increase usually occurs. DHEA and DHEA-S decrease with aging in both men and women [Vermeulen et al., J. Steroid Biochem. 7, 1-10 (1976); Vermeulen et al., J. Clin. Metab. Metab. 54: 187-191 (1982)]. Indeed, at age 70, serum DHEA-S concentrations reach approximately 20% of their peak values ​​and may decrease to 95% at age 85-90. Migeon et al., J. Clin. Metab. Metab. 17, 1051-1062 (1957)]. 70-95% reduction of DHEA-S adrenal formation during aging dramatically reduces androgens and estrogens

Formation in the peripheral target tissues, resulting in a significant reduction of sex steroid-induced biochemical and cellular functions.

In men, in addition to a decrease in DHEA-S formation, a decrease in testosterone concentration in the seminal vein has been observed [Giusti et al., Exp. Gerontol. 10: 241-245 (1975)] and in plasma (Lewis et al., Acta Endocrinol. 82, 444-448 (1976); Zumoff et al., J. Clin. Metab. Metab. 54, 534-538 (1982)] after reaching the age of 60-70 years. However, these data have been the subject of debate (Nieschlag et al., 1982, J. Clin. Endocr. Metab. 55, 676-681). After Climacteria, testosterone concentrations in women are lower than those of reproductive age (MG Forest in Physiological changes in circulating androgens) Androgens in Childhood, MG Forest, Karger Basel, pp. 104-129 (1989). )].

The skin is an essential site for the formation of sex steroids; Skin function is known to be controlled by sex steroids. Sex steroids may act directly on the skin or stimulate the secretion of growth hormone and prolactin by the anterior pituitary gland. It is known that, in the absence of growth hormone, there is indeed atrophy of the skin, probably due to a secondary (secondary) reduction in the secretion of insulin-like growth factor (IGF-1). It is also known that serum concentrations of growth hormone (GH) and insulin-like growth factor (IGF-1) in both men and women are ιί

-4csökkennek.

Plasma concentrations of DHEA-S have been suggested as a predictor of osteoporosis [Nóráin et al., J. Clin. Metab. Metab. 60: 651-657 (1985); Deutsch et al., Int. J. Gynecol. Obstet. 25: 217-220 (1987)]. Indeed, it has been observed that serum DHEA levels in individuals with osteoporosis are significantly lower than in healthy subjects [Nordin et al., J. Clin. Metab. Metab. 60: 651-657 (1985)]. Low DHEA-S levels are likewise accompanied by low tissue D4-dione levels. Because D4-dione is the precursor of the instinct, which is the major source of estradiol in postmenopausal women [Marshall et al., Clin. Endocrinol. 9, 407 (1978)], these secondary, low estrogen levels are also likely to play a role in osteoporosis (Nordin et al., 1981, Chain 2, 277).

Alternatively, low DHEA-S serum levels, which reduce the production of androgens in peripheral tissues, may also result in reduced bone formation, a feature of postmenopausal osteoporosis [Meunier et al. histological heterogeneity and treatment of osteoporosis), DeLuca et al., University Park Press, Baltimore, 293. p.], Deutsch et al., Int. J. Gynecol. Obstet. 25, 217-222 (1987)], there was indeed a significant correlation between serum DHEA-S and androstenedione levels and osteoporosis after climacteria. ; however, there was no correlation between serum estrogens and bone density, suggesting that androgens are more important in post-climacteric bone loss.

Studies in animals have shown that deficiency of androgens induces bone poverty, whereas administration of testosterone increases the total amount of bone (total amount) [Silverberg and Silberberg, 1971; see Finkelstein et al., Ann. Int. Med. 106: 354-361 (1987)]. In rats, testicular eruption causes osteoporosis within two months [Winks and Félts: Calcif. Tissue, Slit. 32, 77-82 (1980); Verhas et al., Calcif. Tissue Slit. 39: 74-77 (1986)].

As noted above, it has been observed that adrenal androgen levels decrease during osteoporosis [Nordin et al., J. Clin. Metab. Metab. 60, 651 (1985)]. Further, it has been shown that post-menopausal women increase their androgen levels to protect against accelerated bone loss [Deutsch et al., Int. J. Gynecol. Obstet. 25: 217-222 (1987); Aloia et al., Arch. Int. Med. 143: 1700-1704 (1983)]. Consistent with this role of androgens in postmenopausal symptomatic menopause, urinary concentrations of androgen metabolites are lower than in control control animals; and in patients with osteoporosis, a significant reduction in conjugated dehydroepiandrosterone (DHEA) has been observed [Holl and White, Acta Med.

Hung. 20, 133 (1964); Urist and Vincent, J. Clin. Orthop. 18

-6199 (1961); Holló et al., Acta Med. Hung. 27, 155 (1970)]. It is believed that post-climacteric osteoporosis is a consequence of low levels (concentrations) of estrogens on the one hand and of androgens on the other (Holló et al., Chain, 1357 (1976)). Because aging (and osteoporosis) is accompanied by a reduction in nearly a number of parameters, and each tissue responds (or does not respond) differently depending on the steroidogenic enzymes present, DHEA levels (concentrations) prior to the discoveries of DHEA treatment of bone cells described below ) and bone loss could not be established.

Regarding the mechanism of both estrogen and androgens' likely role in osteoporosis, the presence of estrogen receptors in osteoblasts (osteoblasts) (Komm et al., Science 241: 81-84 (1988)); Eriksen et al., Science 241: 84-86 (1988)] and the presence of androgen receptors (Colvard et al., Proc. Natl. Acad. Sci. 86: 854-857 (1989)] may explain the increase in bone resorption observed after estrogen and androgen depletion.

Although rapid bone loss occurs in women during the onset of climacteria, in men, bone loss can be observed at about 65 years of age [Riggs et al., J. Clin. Invest. 67: 328-335 (1987). Significant bone loss is observed in men around the age of 80, accompanied by hip, spine and wrist fractures. many

- Study suggests that osteoporosis in men is a clinical manifestation of androgen deficiency [Baran et al., Calcif. Tissue Slit. 26: 103-106 (1978); Odell and Swerdloff (1976) West J. Med. 124: 446-475; Smith and Walker, Calcif. Tissue Slit. 22 (Suppl.), 225-228 (1976)].

Postmenopausal women treated with nandrolone increased mineral content of cortical bone (Clin. Orthop. 225, 273-277), but 50% of patients experienced androgenic side effects. These data are noteworthy because, although most therapies are limited to stopping this bone loss, an increase in bone mass has been observed with the use of nandrolone as an anabolic steroid. Men with hypogonadism have been shown to have similar increases in bone formation using androgens [Baran et al., Calcif. Tissue Slit. 26, 103 (1978)].

The decline in serum levels of DHEA-S and DHEA with age suggests an interesting possibility that low levels of DHEA and DHEA-S may be associated with breast cancer and cardiovascular disease. Indeed, a number of studies indicate that sub-normal concentrations of DHEA are associated with a high risk of breast cancer (Bulbrook et al., Chain 2: 395-398 (1971); Rose et al., Eur. J. Cancer 13, 43-47 (1977); Thijssen et al., J. Steroid Biochem. 6: 729-734 (1975); Wang et al., Eur. J. Cancer 10: 477-482 (1974); and Gomes et al., CR Acad. Sci. Paris 306. 261-264 (1978); Brownsez et al., Eur. J. Cancer 8: 131-137 (1972). It has been observed that in breast cancer • ·•••••••••••• il

-8 women in the urine of androsterone and etiocolanolone

low levels of two metabolites of DHEA (Bulbrook et al., Chain 2, 1238-1240 (1962); Cameron et al., Br. Med. J. 4: 768-771 (1970). Bulbrook et al., Chain 2, 1235-1240 (1962) reported that women with primary operable breast cancer had lower than normal urinary concentrations of 11-deoxy-17-ketosteroids (mainly from DHEA-S and DHEA). ; suggests that reduced separation of DHEA and DHEA-S precedes the development of breast cancer.

The main options for treating advanced breast cancer relate to inhibiting the action and / or formation of estrogens. The role of estrogens in promoting the growth of estrogen-sensitive breast cancer is currently accepted [Lippman, Semin. Oncol. .10, (Suppl. 4), 11-19 (1983); Sledge and McGuire, Cancer Gap. 38: 61-75 (1984); Witliff: Cancer 53: 630-643 (1984); Poulin and Labrie, Cancer Gap. 46: 4933-4937 (1986)].

DHEA (450 mg / kg three times a week) specifically delayed the appearance of breast tumors in C3H mice genetically bred to develop breast cancer [Schwarz, Cancer Sl. 39, 1129-1132 (1979)]. Furthermore, men with low serum DHEA levels have been shown to be at greater risk of developing bladder cancer [Gordon et al., Cancer Slit. 51: 1366-1369 (1991).

United States Patent Application Serial No. 07 / 785,890, filed Nov. 4, 1991, relates to a method of treating breast cancer and endometrial cancer in susceptible warm-blooded animals; this procedure involves the removal of ovarian hormones by surgery (by ovariectomy) or chemically (or by an LHRH agonist such as [D-Trp].⁶, des-Gly-NH 2¹⁰] With LHRH-ethylamide or using an antagonist) as part of combination therapy. Anti-estrogens, androgens, progestins, inhibitors of sex steroid formation (particularly 17B-hydroxy steroid dehydrogenase or aromatase-catalyzed sex steroid production, and the role of prolactin secretion, growth hormone secretion and ACTH secretion) are described. The equivalent of this application is disclosed in PCT International Application Publication No. WO 90/10462.

United States Patent Applications Nos. 07 / 724,532 and 07 / 900,817, filed June 28, and June 24, 1992, respectively, and June 24, 1992, relate to a method for administering low-dose androgenic compounds to breast cancer, prevention and treatment of endometrial cancer, osteoporosis and inflammation of the endometrium. The 1992 application is disclosed in International PCT Application WO 93/00070.

Recent in vitro studies report the relative effects of an androgen proliferation on the proliferation of the estrogen-sensitive ZR-75-1 human breast carcinoma cell strain [Poulin et al. induced cell proliferation, Breast Cancer Slit. Treatm. 12: 213-225 (1989). As mentioned above, Poulin et al., Breast Cancer Slit. Treatm. 12, 213-225 (1989)] found that androgens inhibit the proliferation of cells of the human breast carcinoma ZR-75-1 and that the inhibitory effect of androgens is additive to that of an anti-estrogen. The inhibitory effect of androgens on the proliferation of ZR-75-1 human breast carcinoma cells in vivo in nude mice has also been observed (Dauvois and Labrie, Cancer Slit. 51: 3131-3135 (1991)].

It has been suggested that DHEA has an effect on obesity, diabetes, atherosclerosis, chemical-induced breast cancer, skin cancer and colon cancer (prevention) as well as autoimmune diseases, fatigue, muscle loss, longevity and connective tissue disease Clin. Endocrinol. Metab. 59: 551-555 (1984); Regelson: Ann. NY Acad. Sci. 521: 260-273 (1988); Gordon et al., Adv. Enzyme Reg. 26: 355-383 (1987); Schwarz: Adv. Cancer Slit. 51: 391-423 (1988); Barett-Connor et al., New Eng. J. Med. 315: 1519-1524 (1986).

Schwarz observed in aging Sprague-Dawley rats (Kent: Geriatrics 37: 157-160 (1982)) that their body weight was reduced from 600 g to 550 g without change in food intake due to DHEA administration. Schwarz observed (Cancer 39, 1129-1132 (1979) that DHEA-treated C3H mice (at 450 mg / kg three times a week) were present).

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They gained significantly less weight and reached a longer age than the control animals, had a lower fat content and higher activity. Their weight loss was achieved without limiting their appetite and nutrition. Furthermore, DHEA was able to inhibit weight gain in animals bred to become obese at adulthood (Kent: Geriatrics 37: 157-160 (1982)).

Dietary DHEA has been shown to be an effective anti-hyperglycemic agent for diabetes in mice suffering from a hereditary syndrome of obesity glucose intolerance (Coleman et al., Diabetes 33: 26-32 (1984)).

DHEA reduced the incidence of atherosclerosis in cholesterol-fed rabbits. Gordon et al., J. Clin. Invest. 82: 712-720 (1988); Arad et al., 1989, Arteriosclerosis 9: 159-166. DHEA-S, also present in high concentrations in serum, has been reported to be protective against death from cardiovascular disease in humans (Barrett-Connor et al., N. Eng. J. Med. 315: 1519-1524 (1986)). Circulating concentrations (levels) of DHEA and DHEA-S have been found to be inversely related to cardiovascular mortality (Barrett-Connor et al., 1986, N. Eng. J. Med. 315: 1519-1524), and decrease with the decrease in immune competence [Thoman and Weigle, Adv. Immunol. 46: 221-222 (1989)]. Human testing »

« ·4 »

-12 showed that there is an inverse relationship between fetal DHEA-S serum concentration and serum concentration of low density lipoproteins (LDL) (Parker et al., Science 208, 512 (1980)).

In healthy subjects, 1.6 g of DHEA was orally administered daily for 28 days in a placebo-controlled trial. DHEA serum levels increased 2.5-3.5-fold in the DHEA-treated group, while total serum cholesterol levels and serum LDL cholesterol decreased by 7.1 and 7.5%, respectively. Nestler et al., J. Clin. Endocrinol. Metab. 66: 57-61 (1988)]. Body fat was found to decrease on average by 31% in 5 out of 5 men treated with DHEA without a change in total body weight; it suggests that the muscle mass has increased properly.

Drucker et al., J. Med. Clin. Endocrinol. Metab. 35. 48 (1972); Buster et al. J. Obstet. Gynecol. 166, 1163 (1992) and Wella et al., J. Clin. Endocrinol. Metab. 71, 1259 (1990)] also provided DHEA orally to humans. They observed that^VY mutant mice (Yen et al., Lipids 12: 409-413 (1977)) and Zucker rats (Cleary and Zisk, Fed. Proc. 42, 536 (1983)]. The appearance of C3H mice treated with DHEA appeared to be younger than that of controls [Schwarz: Cancer Slit. 39, 1129-1132 (1979)].

DHEA concentrations in the brain are 6.5 times higher than the corresponding plasma concentrations [Lacroix et al., J. Steroid Biochem. 28: 317-325 (1987). DHEA and DHEA-S improve memory in aging mice [Flood and Roberts: Brain Slit.

448th 178-181 (1988)]. It has also been found that serum concentrations of DHEA-S in subjects with Alzheimer's disease are on average 48% lower than in control subjects of the same age (Sunderland et al., 1989, Chain 570). As described above, chronic administration of DHEA in the diet has been shown to increase age by delaying the development of certain diseases in certain animal species.

U.S. Patent No. 4,496,556 discloses the use of DHEA or derivatives thereof for the treatment of dry skin by topical administration. Only local effects on the sebaceous glands are reported in this communication and no systemic effect has been observed.

U.S. Patent 4,542,129 discloses a topical formulation for treating dry skin of patients; a combination of DHEA and / or derivatives thereof, a keratolytic agent and a non-toxic dermatologically acceptable carrier.

British Patent No. 1,246,639 describes the preparation of esters of dehydroepiandrosterone; these are active ingredients for the treatment of pre- and post-climacteric conditions, tachycardia and headaches.

One problematic aspect of using DHEA in humans is the need for high doses, obviously because most of the compound is degraded in the liver after oral administration before it can enter the bloodstream.

• ·

.............. ί

-14It is known that the efficacy of the delivery (delivery, delivery) of certain drug agents can be improved by using pharmacologically inactive derivatives which are converted in vivo to the active compounds by spontaneous or enzymatic reactions [see generally H. Bundgaard, Design and application of prodrugs. In A textbook of Drug Design and Development (Prodrugs); in the Handbook of Drug Design and Development; ed. P. Krogsgaard-Larsen and H. Bundgaard; Harwood Academic Publishers GmfH, Chur, Switzerland, 113-191. p. (1991)]. For example, Druzgala et al., J. Med. Steroid Biochem. Molec. Biol. 38: 149-154 (1991)] discloses drug precursors of glucocorticoids. Bodor et al., U.S. Patent No. 4,213,978 and German Patent Application DE 2,948,733 disclose the use of thiazolidine derivatives of progesterone as topical active ingredients. DR Friend, Critical Reviews of Therapeutic Drug Carrier Systems, Vol. 7 (2), pp. 149-186. p. (1990)] discloses percutaneous absorption of prodrug derivatives of estrogens and progestins. Further information on percutaneous absorption of active pharmaceutical ingredients can be found in Skin Permeability, H. Schaefer et al., Springer-Verlag, Berlin. Heidelberg, New York, p. 896 (1982).

• ·

Currently, low dose estrogen therapy is currently the standard procedure for the elimination of vasomotor symptoms, urogenital atrophy, osteoporosis, and other symptoms related to climacteria in women around or in chimera. in The Climactic, ed. HJ Buchsbaum, Springer-Verlag, New York, 77-84. p. (1983)]. Further chapters of this book provide detailed information on Climacteria and Therapy. Obviously, these approaches, limited to estrogen replacement therapy, whether related to or not related to progesterins, do not restore the balance between estrogens and androgens naturally occurring in the target tissues through their conversion to active metabolites of DHEA.

Certain esters of DHEA 3-position have previously been described in the literature [Riva et al., J. Org. Chem., 1989, 54, 3161-3164; Parish and Christrakom, Synth. Commun. 15: 393-399 (1985); Romanian Patent Document RO 66924B; Jaros and Zamojski, Tetrahedron 38: 1453-1456 (1982); Heublin et al., 1982, Z. Chem. German Patent Publication No. DE 2 534 914; Khaidem and mun-

<td colspan="2">Wellman and his colleagues:</td><td>Indian J.</td><td>Chem.</td><td>Sect</td><td>. B. 27B, 850-851</td><td> (1988) ;</td>

<td>Pettit</td><td>and</td><td colspan="2">in J.</td><td>Org.</td><td colspan="2">Chem. 52: 3573-3578 (1987);</td>

<td>Hanson</td><td>and</td><td>Reese: J.</td><td>Chem.</td><td>Soc.</td><td>Perkin Trans. 1</td><td> 647-649</td>

(1985); European Patent Application No. 84-105741;

Heublein et al., Acta Polym. 35: 673-677 (1984);

li

-16Severs et al., 1982, J. Med. Chem. 25: 1500-1503; Yamashita and Kurosawa: Agric. Biol. Chem. 39, 2243-2244 (1975); Japanese Patent Application JP 50005372; and Pohlmann et al., Mol. Cryst. Liquid Cryst. 13: 243-254 (1971)].

J. Pharm. Sci., 73, 1643-1645 (1984), discloses that alkanesulfonates of DHEA inhibit the activity of the enzyme glucose-6-phosphate dehydrogenase.

British Patent Application GB 1 246 639 and South African Patent Application ZA 6806112 disclose DHEA esters for the treatment of post- and pre-climacteric tachycardia, headaches and other menopausal complaints.

Leszczynski et al., Biochem. Biophys. Acta 1014. 90-97 (1989); 1083: 18-28 (1991)] discloses plasma esterification of DHEA; Katz et al. Disclose its occurrence in MCF-7 [J. Steroid Biochem. 26: 687-692 (1987).

Weisz and Agocs disclose the ethyl carbonate derivative of DHEA [Arch. Pharm. (Weinheim, Ger.), 319: 952-953 (1986)].

Challis and Heap [J. Chromatogr. 50: 228-238 (1970)] and Pinelly and Nair (J. Med. Chromatogr. 4.3, 223-228 (1969)] disclose some of the halogen esters of DHEA.

Although, as detailed above, it is believed that DHEA may play a role in various biological events, its therapeutic use as a therapeutic or prophylactic agent has so far been relatively limited. In the prevention, amelioration or pre-emptive treatment of certain diseases &lt;1

Their role in reversing their progression (aggravation) is not yet fully understood. The present invention describes several new medical applications of DHEA and DHEA-S (or compounds which are transformed in vivo). The invention further provides novel methods of administering these agents which, for example, overcome the drawbacks associated with oral administration.

The invention is summarized below.

One object of the present invention is to provide simple and effective methods for the systemic delivery of sex steroid precursors, such as DHEA, DHEA-S and their analogs, which are converted in vivo to DHEA or DHEA-S.

It is a further object of the present invention to provide new methods for preventing and / or treating climacteria, vaginal atrophy, hypogonadism, decreased sex drive, osteoporosis, skin thickness and cellularity (skin atrophy), urinary incontinence, ovarian cancer and uterine cancer.

It is a further object of the present invention to provide kits and pharmaceutical compositions for use in accordance with the present invention.

It is another object of the present invention to provide a new method of contraception.

It is another object of the present invention to provide novel sex steroid precursors and pharmaceutical compositions containing them.

In one aspect, the invention provides a process

-18 to treat climacteria; the method comprising administering to a patient in need of such treatment an effective amount of at least one sex steroid precursor; this precursor may be DHEA, DHEA-S, or a compound which is converted in vivo to one of the foregoing in combination with an effective amount of estrogen, progestin or both.

According to a further aspect of the invention there is provided a pharmaceutical composition for treating climacteria and other indications as set forth above; this treatment consists of administering at least one sex steroid precursor, which may be DHEA, DHEA-S or, in vivo, one of the compounds to be converted and a combination of either estrogen or progestin or both. One preferred combination consists of a precursor and estrogen. Another combination consists of a precursor and a progestin. In some preferred embodiments, the precursor, progestin and estrogen are combined. The use of estrogen is not beneficial for male patients.

In another aspect, the invention provides an apparatus for treating a climate bacterium; said equipment comprising a first container containing at least one compound which is converted into a sex steroid precursor by DHEA, DHEA-St or any of these in vivo; and at least one additional container in which progestin, estrogen, or both are contained. In a preferred embodiment, the kit comprises three containers so that the precursor, an estrogen and a progestin-19tin are contained in separate containers.

However, if desired, two or more effective components may be present in a single container. A pharmaceutically acceptable carrier or diluent may be included in one or more containers which may contain preservatives and other additives known in the art.

In another aspect, the invention provides a method of treating vaginal atrophy, hypogonadism, decreased sex drive, reduced skin thickness, and skin cell; this treatment consists of administering to a patient in need of such treatment an effective amount of a compound which is converted to a sex steroid precursor by DHEA, DHEA-S, or any of these in vivo.

In another aspect, the invention provides a method of preventing or treating osteoporosis comprising administering to a patient in need of such prevention or treatment a therapeutically effective amount of a compound which is converted to a sex steroid precursor by DHEA, DHEA-S or in vivo with a pharmaceutically acceptable diluent or carrier. added.

In yet another aspect, the invention provides a method of preventing or treating urinary incontinence, which comprises administering to a patient in need of such prophylaxis or treatment an additional pharmaceutically acceptable or diluent acceptable amount of a therapeutically effective amount of a compound converted to a sex steroid precursor DHEA, DHEA-S, or with or without a vehicle.

In yet another aspect, the present invention provides a method of contraception comprising administering to a woman in need of contraception an effective amount of a compound which is converted to a sex steroid precursor by DHEA, DHEAS-S, or in vivo together with a pharmaceutically acceptable diluent or carrier. without dosing. In preferred embodiments, an estrogen and / or progestin is administered together with the precursor as part of a combination contraceptive method.

In another aspect of the present invention, it is possible to provide a pharmaceutical composition comprising a sex steroid precursor which is converted to DHEA, DHEA-St or any of these in vivo and as an additional active ingredient an estrogen and progestin. In some embodiments, both estrogen and progestin are used. In some preferred embodiments, a diluent or carrier suitable for pharmaceutical practice is added to the composition.

It is also possible to provide kits according to the invention: these consist of at least two separate containers, one containing the sex steroid precursor and the other containing estrogen or progestin or both. In some embodiments, three separate containers may be used, one containing at least one sex steroid precursor, the other container containing at least one estrogen, and an additional container containing at least one progestin.

i

-21talmaz. All of the indications herein, which respond to sexual steroid precursors (DHEA, DHEA-S, or any of the compounds which are intravenous in vivo), in some embodiments, also respond to a combination of an estrogen and / or progestin precursor (except in men where estrogen has a negative effect). Thus, kits and pharmaceutical compositions should provide a combination of the above active ingredients that is appropriate to the particular indication for which it is being used and for which combination therapy is being selected. |

In another embodiment, the invention provides a method of preventing ovarian cancer in a woman in need of such prevention; said treatment comprising administering an effective amount of a compound which is converted to a sex steroid precursor to DHEA, DHEA-S, or any one in vivo, with or without a pharmaceutically acceptable diluent or carrier.

Yet another embodiment of the invention provides a method of preventing uterine cancer; this method comprises administering to a woman in need of such prophylaxis a therapeutically effective amount of a compound which is converted to a sex steroid as a sex steroid precursor by DHEA, DHEA-S, or in vivo, with or without a pharmaceutically acceptable diluent or carrier.

In another aspect, the invention provides a therapeutic method for treating reduced or unbalanced levels of sex steroids by:

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delivering an effective amount of a pharmaceutical composition by percutaneous or transmucosal delivery to the outer surface of the skin or mucosa of a patient in need of such treatment, wherein the pharmaceutical composition comprises a carrier wherein at least one sex steroid precursor is DHEA, DHEA-S or in vivo between the compounds which are converted to any one of them is in a dissolved state; wherein the precursor is present in a concentration of at least 7% by weight based on the total weight of the pharmaceutical composition; the vehicle is compatible (compatible) with the skin or mucosa, allowing the precursor to pass through the skin or mucosa; the viscosity of the vehicle is sufficient to hold the precursor in a localized (defined) area of ​​the skin or mucosa without leaving or evaporating the precursor from this area, allowing a substantially complete penetration of the precursor through the localized area of ​​the skin or mucosa as described above. The above method can be used to treat and / or prevent the conditions, climacteric symptoms, and other conditions described above that respond favorably to replacement DHEA levels. These include, but are not limited to, obesity, cardiovascular disease, atherosclerosis, breast cancer, endometrial cancer, muscle mass loss, diabetes, fatigue symptoms, connective tissue disease, and memory impairment.

Another aspect of the invention provides an opportunity. ₍f ··· · * «* ·« «4

-23 for the preparation of a pharmaceutical composition for the transdermal or mucosal delivery of this pharmaceutical composition comprising a vehicle and, when dissolved therein, at least one sex steroid precursor compound (s) DHEA, DHEA-St or in vivo; wherein the precursor is present in the composition in a concentration of at least 7% by weight based on the total weight of the pharmaceutical composition; and the vehicle is compatible with the skin or mucosa, allowing the precursor to pass through the skin or mucosa; and the viscosity of the vehicle is sufficient to retain the precursor in the localized area of ​​the skin or mucosa without being displaced or evaporated for an appropriate period of time which allows substantially complete penetration of the precursor through the localized area of ​​the skin or mucosa.

In another aspect, the invention provides novel compounds (and pharmaceutical compositions containing them) having the structure of Formula I wherein

X is hydrogen; RC-, RCO₂CHR^the- or R^bsalt₂a group of the general formula;

R is hydrogen; straight or branched alkyl; straight or branched alkenyl; straight or branched alkynyl; aryl, furyl; straight or branched alkoxy; straight or branched alkenyloxy; straight or branched alkynyloxy; aryloxy, furyloxy; or a halogenated analogue thereof;

R^the is hydrogen or C 1-6 alkyl; and

R^b represents a hydroxyl group (or salts thereof); methyl, phenyl or p-toluyl;

Y is a bivalent, substituted or unsubstituted group of formula (a) wherein Z is oxygen or sulfur; and Y, together with the carbon atom to which it is attached, forms a saturated five-membered ring.

Another class of novel compounds is represented by the formula (II), wherein

R^c C3-C20 linear or branched alkyl; C 3-20 straight or branched alkenyl; C 3-20 straight or branched alkynyl; aryl; or a halogenated analogue thereof.

A further class of novel compounds is represented by compounds of formula III wherein:

R^d is hydrogen; straight or branched alkyl; straight or branched alkenyl; straight or branched alkynyl, aryl, furyl; straight or branched alkoxy; straight or branched alkenyloxy; straight or branched ·· »·» · · «« ·· *

-C25 alkynyloxy; aryloxy, furyloxy; or a halogenated analogue thereof; and

R^the represents a hydrogen atom or a C 1-6 alkyl group.

A further class of novel compounds includes compounds of formula IV wherein R³ represents a hydroxyl or sulfate group;

R^e is hydrogen, benzyl, aryl; straight or branched alkyl; straight or branched alkenyl; straight or branched alkynyl; and

Z represents oxygen or sulfur.

One embodiment of the present invention provides a method for compensating for the reduced separation of the sex steroid precursors DHEA and DHEA-S; these precursors are secreted by the adrenal glands in decreasing amounts as they age. This method comprises administering DHEA, DHEA-St, or an analogue thereof in vivo to any one of these, which is capable of compensating for the consequences of reduced separation of DHEA and DHEA-S produced by aging adrenal glands without undesirable side effects. .

In our view, the methods of the invention are suitable for both prophylactic and therapeutic use. Serum Concentrations, Assets reported in this manual.

-26s (kits) and preparations are suitable for any of the above purposes.

A further aspect of the invention provides a transdermal device that provides

(a) a surface which comes into proper contact with the human skin;

b) an accessory that holds this surface in the localized area of ​​the skin to which the transdermal device is applied;

c) a storage unit which is in fluid communication with the surface and which comprises a pharmaceutical composition comprising a carrier and the active ingredient dehydroepiandrosterone, dehydroepiandrosterone sulfate or an analogue of any of these, which is converted in vivo; as well as

d) comprises an accessory means for delivering the pharmaceutical composition from the storage unit to the localized area of ​​the skin through the surface.

It will be appreciated that compounds which are transformed into a sex steroid precursor by DHEA, DHEA-S, or any in vivo condition (e.g., prodrug forms of DHEA or DHEA-S) will be used in all indications suggested herein for use in the present invention. All cause an increase in DHEA serum concentrations. Because DHEA-S is a natural precursor to DHEA, DHEA-S (and its prodrug forms of DHEA or DHEA-S) can be used in place of DHEA in all of the different indication areas described above where DHEA can be used. For these pro • · · · · · A ·· &lt;«« «« · * * · · ί

Administration of -27drug forms or DHEA-S results in the desired increase in DHEA levels.

Below is an explanation of the figures.

1-3. Figures 1 to 5 show the increase in bone density of rats after 9 months of DHEA (EM-760) treatment. Figure 1 shows the total bone mass density of DHEA-treated, intact, and ovariectomized rats for 9 months compared to untreated control rats. Figure 2 illustrates the same comparison in terms of lumbar spine density; Figure 3 shows the same comparison of femoral mass density. In all cases, except for the intact control group, measurements were made 9 months after ovariectomy and / or dermal DHEA administration. These figures illustrate the value of the present invention in the treatment or prevention of osteoporosis.

Figures 4 and 5 illustrate the efficacy of DHEA in the prevention of contraception and in the treatment or prevention of ovarian or uterine cancer. Figure 4 illustrates the reduction in rat ovarian weight by administering DHEA percutaneously twice daily for 6 months in a dose of 30 mg twice in 0.5 ml of a solution of 50% ethanol and 50% propylene glycol in about 2 cm² size area.

Figure 5 illustrates the histological observation of the ovaries of healthy control rats (Figure 5a) and of DHEA-treated healthy rats (Figure 5b). DHEA is applied to the animals' back skin 2 cm² 30 mg twice daily, i

··»·»*· · • · · * .. · » • · · · · · «· «« «.·».·· ·

I

-28ί

It was added dissolved in a mixture of 50% ethanol and 50% propylene glycol. 5b. Fig. 3A shows a marked decrease in the number of tertiary and secondary follicles (F), as well as the absence of corpus luteum (CL) and intercellular cells (IC). The magnification is 250 times. Remarkably, the treated rat (Fig. 5c) exhibits interstitial gland atrophy compared to the intact animal (Fig. 5d) at a magnification of 500 times.

Figure 6 illustrates the effect of 1, 3, or 6 months of DHEA treatment on vaginal atrophy. DHEA, dissolved in a mixture of 50% ethanol and 50% propylene glycol at a dose of 30 mg twice daily, was administered to 2 cm of the dorsal skin of rats deprived of their ovaries.² area. Atrophic vaginal epithelium is seen 1 month (Fig. 6a), 3 months (Fig. 6b), and 6 months (Fig. 6c) after castration. Reversed vaginal atrophy was observed in rats treated with DHEA as shown in Figs. 6d, 6e. and 6f. Figures 4A-5B show vaginal epithelium after 1, 3, and 6 months of DHEA treatment (200x magnification). Thus, histopathological examination showed that rats treated with DHEA showed proliferation and enhancement of the vagina, and reversal of vaginal atrophy.

Figure 7 shows the histopathology of the skin of male rats treated with DHEA for 3 and 6 months, respectively; DHEA was topically applied to the back skin of the animals 2 x 2 cm² territory. 7b. and 7c. Fig. 4A shows mild to moderate hypertrophy and hyperplasia of the sebaceous glands on the dorsal skin; it is also seen on the skin of intact rats treated for 3 months with 7e. and 7f. FIG; and in animals treated for 6 months, as shown in FIG. and 7f. Figures; these are illustrated in FIG. and 7d. In Figures 1 to 4, we compared intact controls with 9V * • 9 · * 4 · 9 · *. 7d. FIG.

Figure 8 shows the histology of the skin of male rats treated with DHEA for 3 and 6 months; DHEA was topically applied to the back skin of the animals 2 x 2 cm² territory. Observed in Fig. 8b. and 8c. Figure 3B shows mild to moderate hyperplasia of the sebaceous glands on the dorsal skin; hyperplasia of the sebaceous glands was also observed in rats (3) (Figures 8b and 8e) and 6 months (8c and 8f) who had been castrated on the abdominal skin (Figures 8e and 8f). This was compared with castrated controls (Figures 8a and 8d). Noteworthy is the increase in wiring tension (elongation) (D) and acinar (A) per hair follicle (magnification 100x).

Figure 9 shows the effect of DHEA treatment on the dorsal layer of the dorsal skin (area of ​​topical application of DHEA) in male castrated rats after 3 months (Figure 9c) and after 6 months (Figures 9d and 9f). Castrated, untreated rats were used as controls (Figures 9a, 9b and 9e). In treated rats, the thickness and cellularity (cellular consistency) of the skin layer increased (Figures 9c, 9d, 9f) (100x magnification in Figures 9a, 9b, 9c and 9d, 9e and 9f). 500x magnification).

7-9. Figures 3 to 5 illustrate that topical application of DHEA to the dorsal skin leads to an increase in the thickness and cellular character of the loose skin; it is an effect that can prevent or treat skin atrophy (skin atrophy is a condition in which, among other things, collagen connective tissue loss occurs. »· * • · · · · · ··· ·· ···« 999 &lt;

-30jellemző). The effects of DHEA are also seen in the size of the sebaceous glands, and we believe that the growth of the sebaceous glands is stimulated by the androgenic compounds formed by DHEA as a precursor.

Figures 7 and 8 show that topical application of DHEA (30 mg in a mixture of 50% ethanol and 50% propylene glycol) was applied to the back skin 2 x 2 cm for 3 and 6 months, respectively.² It causes mild-to-moderate hypertrophy and hyperplasia (proliferation) of the sebaceous glands in both the dorsal and abdominal areas, suggesting a generalized effect of skin treatment. This effect can be observed in both intact and castrated animals; pronounced atrophy following castration is completely eliminated (prevented) by DHEA treatment. The reduction in the size of the sebaceous glands following castration can be compared to atrophy during aging.

As shown in Figure 9, the effect of DHEA in both intact and castrated animals is accompanied by a significant increase in the thickness and cellularity (cellular nature) of the cutaneous layer. As collagen is an important component of the cutaneous layer of the skin, current data show that the elimination (correction) of skin atrophy can be achieved, at least in part, by enhanced formation of collagen tissue.

The contraceptive efficacy of DHEA is indicated by the topical application of DHEA (30 mg in 50% ethanol / 50% propylene glycol) twice daily to 2 cm²histopathologic lesions in the area, showing inhibition of ovulation. The most important changes observed are ovarian atrophy (Fig. 4) and histopathological lesions (Fig. 5), which indicate the absence of ovulation cycles. The number of secondary and tertiary follicles decreased significantly, and the corpus luteum is absent, indicating a lack of ovulation. In addition, the histopathologic signs of ovarian deficiency in ovaries are accompanied by the disappearance of the cyclic histopathological changes normally seen in the rat endometrium during the estrogen cycle. These histological changes suggest that DHEA treatment has a contraceptive effect.

The invention will be further elucidated in the following non-limiting description of preferred embodiments.

To select patients for whom the treatments described below may be useful, measurement of serum levels of DHEA and its metabolites may be performed as described by Belanger et al., Steroid Formation, Degradation and Action in the Peripheral, Normal and Neoplastic Tissues. in normal and cancerous tissues), ed. H. Bradlow et al., Ann. NY Aced. Sci. 586: 93-100 (1990); Haning et al., J. Clin. Endocrinol. Metab. 72, 1088 (1991)]. See also Labrie et al., Endocrinology 123: 1412-1417 (1988). Serum IGF-1 levels can be measured as described by Furlanetto et al. Clin. Invest. 60, 648 (1977)]. According to the present invention, after determining DHEA deficiency, DHEA or its analogs are preferably administered in a dose sufficient to provide a serum DHEA concentration of 4 to 10 micrograms per liter, preferably 4

-32 to 7 micrograms and keep it at that level. As discussed below, higher concentrations are required in some indication areas.

In some preferred embodiments, the serum concentration is between 5 and 7 or between 6 and 7 micrograms per liter. In contrast, concentrations of up to 13 micrograms / liter (e.g., 7 to 13 micrograms / liter) are preferred for contraception or for prevention of ovarian or uterine cancer. The preferred doses detailed below may be increased as necessary to produce higher serum concentrations; for example, they may be increased by about 30% depending on the individual response observed by the caregiver. When DHEA was preferably administered by the percutaneous (transdermal) or mucosal route, it was found that DHEA was very efficiently absorbed into the blood and thus increased serum levels. For example, if a Glaxal cream (available from Glaxal Canada Limited) containing Glaxal as a carrier and 10% by weight of DHEA, based on the total weight of the formulation, is used in an amount of 100 milligrams of active ingredient, such as DHEA, twice daily area per 100 cm 2, an average patient is likely to respond with an increase in serum DHEA of about 0.7 micrograms per liter of body weight. The dosage applied may be increased or decreased in a known manner by modifying the site to which the wash solution or ointment is applied; or surface you • ·

Changing the size of the circumference at which it is applied; or by adjusting the concentration of the active ingredient; or by changing the vehicle. For example, if the surface area is increased, the dose of the active component will generally increase if the concentration of the active component remains constant. Similarly, as the concentration of the active ingredient in the carrier matrix increases, the dose applied increases, and as the concentration decreases, the dose administered decreases. The dose to the bloodstream (bloodstream) also varies in a known manner depending on the area of ​​the body where the transdermal permeation system is applied to the skin. Changing the vehicle may also modify the dose administered in a known manner. Preferably, serum DHEA concentrations are measured prior to commencement of treatment, and a dose is selected that provides a rapid range of serum DHEA concentrations within the range of 4 to 10 micrograms per liter, or 7-13 micrograms per liter for the above-mentioned higher dose indications. lifts. Thereafter, the patient is monitored for both onset of symptoms and monitoring of DHEA levels to confirm that the desired target serum concentration and symptomatic relief (symptom resolution) has been achieved. DHEA is then maintained in the circulation at a constant concentration. For an average post-climacteric patient, for example, this dose is equivalent to applying 400 mg of active precursor as part of a 10% formulation of Glaxal so that the dosage is 400 cm² twice daily (50 kg train-34). If oral administration is chosen, 800 mg twice daily should be administered.

In accordance with the present invention, DHEA, DHEA, and / or any of the compounds are administered to treat and / or prevent climacteric symptoms, vaginal atrophy, skin atrophy, hypogonadism, decreased sex drive, osteoporosis, urinary incontinence (inability), ovarian cancer or uterine cancer. . In addition, DHEA, DHEA-S (or analogues) administered transdermally (transdermally) according to the invention can also be more effective in treating other pathological conditions associated with decreased secretion of DHEA by the adrenal glands during aging, respond (respond) to therapy. Pathologic conditions that are expected to respond to the treatments described herein can be diagnosed in the usual manner. For example, breast cancer can be noticed by the patient himself or may be the result of clinical breast examination (by a doctor) and / or mammography. On the other hand, endometrial cancer is usually observed with biopsy of PAP, smear and / or endometrium. Both types of cancer are well known to those of ordinary skill in the art using standard physical techniques such as scanning bone scan, chest x-ray, skeletal examination, liver ultrasound and scanning (if desired), CAT scanning, MRI and physical examination. by examination - can be diagnosed and evaluated.

The first symptoms of climacteria are hot flashes. The prints are out

-35 space can be further characterized by known methods [see, e.g., The Menopause (climacteric A) ed. HJ Buchsbaum, Spriger Verlag, New York, p. (1983)]. Vaginal atrophy is often indicated by painful intercourse and vaginal infections. Vaginal atrophy, hypogonadism, and decreased sex drive are well known. For the diseases mentioned above, see, for example, SG Korenman, Sexual Dysfunctions, in Williams Textbook of Endocrinilogy, JD Wilson et al., WB Saunders Co., Philadelphia, 1033-1048. p. (1992).

Bone density can be measured by standard techniques well known to those skilled in the art, such as QDR (quantitative digital radiography), dual-photon absorption, and computer assisted tomography. Useful parameters for bone formation and bone resorption: plasma and urinary calcium and phosphate concentrations, plasma alkaline phosphatase, calcitonin and parathyroid hormone levels, and urinary hydroxyproline and calcium / creatinine ratios.

The aging process, especially in individuals over the age of 50, is often accompanied by loss of collagen or connective tissue in the skin. This is clearly evident from skin wrinkling and / or loss of elasticity.

In accordance with the present invention, osteoporosis or other osteoporosis and other diseases that can be treated by activating the androgen receptor are within the scope of the present invention.

-36 can be treated or prevented. The invention can help prevent cancer of the breast, ovary and endometrium.

Normal body weight ranges are well known to those skilled in the art; cholesterol and lipoproteins can be measured routinely by standard procedures [Nestler et al., J. Clin. Endocrinol. Metab. 66, 57-61 (1988) and quotations therein].

Skin condition can be assessed by eye (eye), palpation, more precisely by prick biopsy and standard histological examination.

The traditional main mechanism currently adopted for female contraception, associated with estrogen administration, is that an increase in estrogen blood circulation levels reduces the release of LHRH from the hypothalamus, which in turn reduces the release of LH from the pituitary gland. The reduced LH secretion thus reduces the function of the ovaries and in particular the ovulation. The addition of progestin controls the proliferation of the uterine lining and changes the separation of the vagina and cervix, creating an unfavorable environment for sperm reception and conception.

DHEA is used in place of estrogen according to the invention (although estrogen may be added in some of the embodiments described below). According to the present invention, DHEA is the estrogen required for contraception, while simultaneously and desirably providing elevated concentrations of androgens that bring about contraception.

<img file="HUT73241A_D0002.tif" />

-37, because androgens inhibit the separation of LHRH and LH, especially in women around the age of climacteria (and in women of post-climacteric age when contraception is no longer needed), these androgens can provide much needed stimulation of bone formation and provide resistance to bone loss. . Estrogens produced from dosing DHEA also contribute to the reduction of bone loss. When used as a replacement for DHEA (by substitution of DHEA), estrogens are produced by natural processes in the same tissues where estrogens and androgens are needed, and which normally convert DHEA into estrogens and androgens. The relative ratios of estrogen to androgen in each specific tissue remain essentially at natural levels.

As with other uses described herein, DHEA-S or DHEA or DHEA-S prodrug forms may be used in place of DHEA. As the contraceptive method described results in a reduction in ovarian function, the production of estrogen and progesterone in the ovaries is also reduced. Thus, it is preferred to administer a progestin (e.g., medroxyprogesterone acetate, megoestrole acetate, noretinodrel, L-norgestrel) as part of a method of contraception to prevent uterine hypertrophy; androgenic progestins are preferred. The progestin can be administered in the DHEA-containing pharmaceutical composition or separately. In some embodiments, the progestin may be administered alternately every month for 12-14 days, or every few months.

-38 (e.g. every 2-4 months) for 12-14 days or continuously. The progestin may be administered in the dosage range currently used, but lower doses are preferred for the reasons explained below.

Due to the reduced production of estrogens in the ovaries, estrogen may be added to contraceptive therapy; however, DHEA itself is converted into estrogen in many tissues, and thus exogenous estrogen can be minimized by practicing the invention. When used in a contraceptive method, the preferred dose of estrogen administered is an amount which achieves 100-200 nanograms / liter or equivalent. The weight ratio of estradiol to DHEA is preferably in the range of 1000 to 25000, preferably 2000 to 15000, in particular 3000 to 12000. As with progestin administration, estrogen may be administered as part of a pharmaceutical composition containing DHEA (or, optionally, DHEA-St or their prodrug form), or may be administered separately. In some embodiments, both DHEA and progestin and estrogen are administered, either alone or separately, as part of a combination therapy. Combination therapy is effective when, due to the treatment regimen, the blood levels of each active substance are simultaneously increased. This simply requires that the active ingredients be added at a time close enough to each other to achieve increased blood levels of the active ingredients at the same time.

So far, the combination of an estrogen and a progestin contraceptive has not been shown to reduce

-39 the risk of breast cancer (Romiev et al., Cancer 66: 2253-2263 (1990)). These data agree that both estrogen and progesterone are known to have a mitogenic effect on proliferation of mammary epithelial cells, which explains that cell proliferation reaches a peak in the middle of the luteal phase [Masters et al., J. Natl. Cancer Inst 58, 1263-1265 (1977); Anderson et al., 1982, Br. J. Cancer 46: 376-382. Indeed, all mammals of pre-climacteric women using contraceptives do not have the same proliferation rate as untreated women with cycles (Potter et al., 1988, Br. J. Cancer 58: 163-170; Going et al., Am. J. Pathol. 130: 193-204 (1988)].

We have recently discovered that DHEA is preferentially converted to androgens over estrogens. Thus, in accordance with the present invention, this prohormone is used as a contraceptive instead of the current contraceptive in order to desirably reduce the proliferation of breast cells. In fact, androgens inhibit the proliferation of breast cells through two mechanisms; on the one hand, they exert a direct inhibitory effect on breast cells; on the other hand, they inhibit gonadotropin secretion at the level of the hypothalamus and pituitary, resulting in decreased ovarian function (estrogen secretion and consequently estrogen-induced mammary cell proliferation).

In addition to its beneficial effects on breast cells, DHEA-induced decrease in gonadotropin secretion decreases

-40i ovarian activity (see Figures 4 and 5) and thus is expected to help prevent ovarian cancer and uterine cancer. Consequently, increased gonadotropin secretion prior to climacteria is improved by DHEA administration. Thus, according to the invention, DHEA can be used prophylactically against the development of ovarian or uterine cancer, even in patients at high risk of developing such cancer, even though contraception is not the primary goal.

Prior art prior art oral contraceptives contain significant amounts of sex steroid estrogen.

genes and progestins are added. However, the pathway of the present invention is based on the tissue-specific formation and activity of dominant androgens synthesized from DHEA. Indeed, when peripheral tissues are exposed to the DHEA steroid precursor, these tissues predominantly produce adrogens and some estrogen at the site (s) of action, resulting in a physiological balance between the two categories of sex steroids. DHEA also inhibits gonadotropin secretion, thereby protecting the ovary from overstimulating the presence of high concentrations of gonadotropins, which precede and accompany climacteria. Gonadotropin secretion is largely inhibited by natural androgens from DHEA, thus minimizing the need for estrogens and progestin. This is important because many progestins, including norethindrone and noretinodrel, have potent estrogenic effects [Poulin et al., Breast Cancer Slit. Treat. 13, 265-276 (1989)].

• ·

<img file="HUT73241A_D0003.tif" />

-41 We have further recognized that DHEA, as a steroid precursor, is converted into androgens (and estrogens) in osteoblasts (bone-forming cells). This discovery suggests that DHEA may be used in the treatment or prevention of osteoporosis, in accordance with the present invention, instead of androgen and estrogen. Bone-produced androgens (produced by the conversion of dosing DHEA) stimulate bone formation and reduce bone loss, while estrogens produced from dosing DHEA also contribute to the reduction of bone loss. Thus, significant side effects of conventional androgen therapy can be avoided. For example, androgens currently administered externally can reach many tissues that do not produce and require androgens; thereby causing side effects and disturbing the physiological balance of sex steroids in these tissues. If DHEA (or its prodrug forms or, if desired, DHEA-St) is used instead, DHEA will only be converted to androgens in the tissues by natural mechanisms that perform this localization. Further, the relative ratio of androgens to estrogens produced by DHEA is essentially a normal ratio, rather than an abnormally increased ratio of a single type of sex steroid, if only this type is used.

In a preferred embodiment, DHEA is administered in an amount sufficient to prevent or treat osteoporosis at a dose that maintains substantially normal serum concentrations in young adults: approximately 4-10 l / liter.

-42 micrograms, in some embodiments 4-7 micrograms per liter, e.g. 5 to 7 or 6 to 7 per liter. These concentrations are also desired for other DHEA responsive (reactive) indications described herein, except for contraception and the prevention of ovarian and uterine cancer, where in some embodiments, the preferred dose may be increased to 13 micrograms per liter (as in order to further inhibit the secretion of LH produced in the anterior pituitary gland.

In a preferred form of treatment for climacteria, the object of the present invention is to maintain the blood levels of estrogen and a sex steroid precursor (such as DHEA or DHEA-S) within the normal pre-climacteric parameter values. In most peripheral tissues, the body converts DHEA-St to DHEA. Without wishing to be bound by theory, it is believed that maintaining appropriate concentrations of the precursor enhances the ability of natural enzymes such as 17β-hydroxysteroid dehydrogenase, 3α-hydroxysteroid hydrogenase, aromatase and 5α-reductase. and estrogen production, and to maintain them in the sense that they remain similar to their relative concentrations prior to the climacteric. Thus, according to the invention, it is considered that not only estrogens but also androgens and precursors are in a better balance. In fact, all target tissues have a set of enzymes that are local to the synthesis of androgens and / or estrogens * · ·· ♦ · ί

-43 is desirable for regulation and needs [Labrie: Mole. Cell. Endocrinol. 78, C113-C118 (1991)].

When estrogen is administered in combination with a precursor (e.g. DHEA) according to the invention, it can be administered simultaneously or separately. The second component (precursor) may also be added to an estrogen therapy in progress to effect the combination therapy of the present invention.

All that is required is that both the estrogen and the precursor be administered in sufficient amounts to achieve the desired concentrations of both substances. According to the combination therapy of the present invention, the concentration of the precursor is maintained within the desired range of parameters while at the same time the concentration of estrogen is maintained within the desired range of parameters. When estradiol is used, serum estradiol concentrations are generally maintained within the range of 50 to 300 nanograms / liter, preferably 100 to 200 nanograms / liter, most preferably 150 to 175 nanograms / liter. If other estrogens are used, the serum concentration should be adjusted in a known manner, taking into account the difference in estrogen activity relative to estradiol and the achievement of normal pre-climacteric estrogen levels (estrogen concentrations). For example, when using Mestranol, lower concentrations are required. Appropriate estrogen serum concentrations can also be evaluated based on the resolution of climacteric symptoms. Serum concentrations of the second compound of combination therapy (such as DHEA) are generally between 4 and 10 * · ··

<img file="HUT73241A_D0004.tif" />

ί

-44 micrograms / liter, in some embodiments maintained between 5 and 7 micrograms / liter or 6 and 7 micrograms / liter.

The estrogen is preferably estradiol but may be sodium estrone sulfate or another compound which acts as an estrogen receptor agonist. When administered separately, commercially available estrogen supplements such as Premarin (Ayerst, St-Laurent, Quebec, Canada) may be used. DHEA is a preferred precursor, although DHEA-S and its analogs as described below are particularly effective for the reasons explained below. When DHEA is used, for example, DHEA (pharmaceutical grade) is available from Sigma (St. Louis, Missouri, USA). For the average patient, the oral dose of estrogen required to achieve the desired serum concentrations is 0.3 to 2.5 mg Premarin per 50 kg body weight per day. In some embodiments of the invention, the estrogen may be 17B-estradiol, administered by percutaneous patch, available from Ciba under the tradename Estraderm, at a daily dose of 0.05 to 0.2 milligrams per 50 kg body weight. For average patients, the appropriate dose of DHEA as a sex steroid precursor to achieve the desired serum concentration of the precursor in oral administration is 0.25-2.5 grams (briefly: gram = g; milligram = mg; microgram = μg; nanogram = ng) per 50 kg body weight. ). Other sex steroid precursors are dosed at a dose that is in · · · · · *

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yl

-45vivo conditions depend on the extent of their conversion to DHEA. In addition, the precursor may be administered transdermally (as described below) in sufficient amounts to achieve the appropriate target serum concentration. This correlation is also described in more detail below.

Alternatively, climacteria may be treated with a precursor as described above in combination with a progestin such as medroxyprogesterone acetate (e.g. Provera).^R), which is preferably administered alternately, for example for 12 consecutive days, in a dose of 2 to 10 mg daily, alternating 12 day periods with a 20 day interval at 5 month intervals. Combination therapy using the precursor, an estrogen and a progestin may also be performed, preferably using the doses given for each component herein.

The sex steroid precursor used in the present invention may be administered orally with or without the addition of a solvent or diluent, but if preferably administered transdermally or mucosa, the addition of a vehicle or diluent in a pharmaceutical composition for oral administration is preferred for DHEA or other precursor. and 98% by weight, more preferably 50 and 98% by weight, particularly preferably present in an amount of from 80 to 98% by weight. If the composition also contains estrogen, such as estradiol, its concentration is preferably 0.04-0.4% by weight. The only active ingredient (active ingredient) can be a single precursor moiety! .

i · * »· · * &lt;· ··« «» ·^μ ♦

-46 for example DHEA; however, several precursors and / or analogues thereof may be used (e.g., a combination of DHEA and DHEA-S; or a combination of two or more compounds which are converted in vivo to DHEA or DHEA-S; or combining DHEA with one or more thereof). analogue which is converted in vivo to DHEA). If a combination is used, the sum of all doses of all precursors should be within the dosage range given above for DHEA alone, taking due account of the various molecular weights of the DHEA analogs, such as the molecular weight of the DHEA esters and their conversion rate. Thus, if a DHEA ester is used in place of DHEA, the dose should be increased by a factor corresponding to the molecular weight of the DHEA ester relative to the molecular weight of DHEA. The final criterion for an appropriate dose is the blood level of DHEA: this dose should also take into account individual changes in absorption and metabolism.

The clinician, particularly at the start of treatment, preferably monitors the patient's overall response (response) and serum concentrations of both estrogen and DHEA (relative to the preferred serum concentrations above), and monitors the patient's overall response to treatment, and dose adjustment may be required if the patient's metabolism or response to treatment is abnormal (atypical). One way to do this is to start treatment with DHEA or anaglone alone and then add estrogen if you have estrogen levels · «*« ♦ ·

<img file="HUT73241A_D0006.tif" />

-47 is too low. Many patients can be treated with the precursors of the invention without additional estrogen.

The treatment according to the invention can also be used without any time limit. Except for the above-mentioned indications requiring higher doses, DHEA treatment is expected to simply maintain DHEA levels in a range similar to that found in women before the onset of climacteria (4 to 10 µg / liter liter) or naturally in young adult men. (serum concentration 4-10 µg / liter). Accordingly, prolonged DHEA treatment is expected to result in minimal or no adverse effects. Side effects associated with long-term use of estrogen can be avoided by methods known in the art, for example, by administering a progestin (e.g., medroxyprogesterone acetate) at a daily dose of 2 to 10 mg orally, intermittently (or continuously according to another embodiment).

For the purpose of facilitating the combination therapy of the present invention, any of the indications disclosed herein may be contemplated by pharmaceutical compositions comprising both estrogen and the second active compound (precursor) in a single formulation for simultaneous administration. The composition may be suitable for administration by any conventional route, such as, but not limited to, oral administration, subcutaneous injection or intramuscular injection. In another embodiment, the invention provides a kit (set) which is contained in separate containers₉*, Μ · «« * ♦ ♦ · ♦ ♦ »» »» »» '' '' '

-48 Contains estrogen and the second compound (precursor (s)). In addition to other modes of administration, the second compound and estrogen may also be administered transdermally in accordance with the present invention, as will be explained in more detail below. Based on this, the kit may contain materials suitable for transdermal (transdermal) administration, such as ointments, lotions, gels, creams, sustained release patches. The same procedure applies to progestin. As noted above, estrogens are disadvantageous when combination therapy is used in male patients.

We have discovered that administration of DHEA can be used to treat and / or prevent vaginal atrophy, osteoporosis, skin atrophy, uterine cancer, vaginal cancer, urinary incontinence, hypogonadism, and decreased libido; and improves the overall balance of circulating sex steroids, including estrogens and androgens. We believe that these pathological conditions have so far been unlikely (not recommended) to respond to DHEA treatment. It is contemplated that compounds which are transformed into DHEA, DHEA-S, or in vivo, will be useful in the treatment of all of the diseases listed above.

Methods known hitherto for the systemic administration of DHEA are partly oral and partly injection methods. Because DHEA treatments are often long lasting and even indefinite, repeating injection dosing is very uncomfortable. However, oral administration is relatively unsatisfactory. · ···. :.

··· · ί ··· «··« · d

-49 proved to be the first route of oral administration of DHEA to the liver, which prevents a large proportion of it from reaching the general circulation due to local degradation.

More recently, it has been observed that DHEA is very effectively absorbed systemically in both men and women after application to the skin or mucous membrane (e.g., oral, vaginal, or rectal mucosa). It has been discovered that therapeutically effective doses of DHEA can be administered percutaneously (transdermally) or through the mucosa, thereby avoiding the first passage of the steroid through the liver, which is the result of oral administration, and avoiding inappropriate and inconvenient injection of DHEA.

Accordingly, the present invention provides delivery systems for delivery of DHEA, DHEA-S, and analog compounds that are in vivo transformed into any of these via the skin or mucosa. In our view, these systems are more effective than oral administration because they bypass (shunt) the liver. Furthermore, these systems are much less painful and more comfortable than injections.

When DHEA, DHEA-St, or analogue compounds that are converted in vivo to DHEA or DHEA-S are formulated for transdermal (transdermal) passage, a variety of transdermal delivery systems are known in the art. For example, DHEA may be used as part of an ointment, lotion, gel or cream to rub the patient's skin. The active ingredient (active ingredient) is based on the total weight of the pharmaceutical composition

-50 preferably 7-20% by weight, more preferably 8-12% by weight. Alternatively, the active ingredient may be incorporated into a transdermal patch (the structure of which is known in the art, see, for example, European Patent 0 279 982).

To form an ointment, lotion, gel or cream, the active ingredient is admixed with a suitable vehicle that is compatible with human skin or mucous membrane and enhances transdermal passage of the compound through the skin or mucous membrane. Suitable carriers for this are already known, including but not limited to Klucel HF and Glaxal. Some are commercially available, such as Glaxal raw material (Glaxal Canada Limited Company). Other suitable carriers are described in Roller and Buri (1987, STP Pharma 3 (2), 115-124). The carrier is preferably of a nature in which the active ingredient (s) is (are) soluble at room temperature in the concentration of the active ingredient employed. The vehicle should have sufficient viscosity to retain the precursor in the localized area of ​​the skin or mucous membrane to which the composition is applied, without displacement or evaporation for a sufficient time to permit substantially complete penetration of the precursor through the localized area of ​​skin or mucous membrane induces a measurable desired increase in serum DHEA. The carrier will generally be a mixture of several components, for example a pharmaceutically acceptable solvent

<img file="HUT73241A_D0007.tif" />

-51rek and a mixture of a thickener (thickener). A mixture of organic and inorganic solvents, such as water and an alcohol such as ethanol, may promote hydrophilic and lipophilic solubility.

The desired excipient is such that, when formulated in a formulation containing 10% by weight DHEA and 90% by weight of the vehicle and applied to the abdomen in an amount providing 100 mg of DHEA twice a day, the average concentration of DHEA in an average patient is 50%. It is increased by at least 0.35 Mg / liter body weight. As mentioned above, when used as a vehicle under these conditions, a Glaxal base material allows an increase in serum DHEA concentration of about 0.7 Mg / liter per 50 kg body weight.

The carrier may also contain various additives commonly used in ointments and lotions, which are well known in the cosmetics and medical applications. Examples include perfumes, antioxidants, gelling agents, thickeners such as carboxymethylcellulose; humectants, stabilizers, plasticizers, coloring matter and the like. For the treatment of systemic diseases, the site of application should be modified to avoid excess local steroid concentrations, thereby eliminating the potential for overstimulation of the skin and sebaceous glands by DHEA androgenic metabolites.

In some cases, the precursor may also be administered orally, with the excipients (excipients) used in conventional pharmaceutical practice, e.g.

<img file="HUT73241A_D0008.tif" />

dried lactose and magnesium stearate can be converted into tablets or capsules for oral administration at concentrations providing a daily dose of 0.25 to 2.5 g per 50 kg body weight.

The active ingredient may also be processed in the form of tablets or dragee cores by mixing the active ingredient with solid powdered carriers such as sodium citrate, calcium carbonate or dicalcium phosphate, and binders such as polyvinylpyrrolidone, gelatin or cellulose derivatives, as appropriate. lubricants such as magnesium stearate, sodium lauryl sulfate, Carbowax or polyethylene glycol. Of course, flavor enhancers may also be added to the formulation for oral administration.

Additionally, capsules, for example hard gelatin capsules, and sealed soft gelatin capsules containing a plasticizer or plasticizer, such as glycerol, may also be used as pharmaceutical forms. The capsules preferably contain the active ingredient in particulate form, for example, in admixture with excipients such as lactose, sucrose, mannitol, starch (e.g., potato starch), cellulose derivatives or finely dispersed silica. In soft gelatine capsules, the active ingredient is preferably dissolved or suspended in a suitable liquid, such as vegetable oils or liquid polyethylene glycols.

The concentration of the active ingredient in the ointment, cream, gel or lotion is generally about 7-20% by weight, preferably 8-12%, more preferably 10% by weight (based on the total weight of lotion-53dat, cream, gel or ointment).

Within the preferred range, higher concentrations allow a suitable dosage to be achieved by applying the rinse solution, ointment, gel or cream to a smaller surface area of ​​the skin than is possible at lower concentrations; this allows the free choice of the parts of the body to which the ointment or lotion is to be applied, for example, it is well known that a compound capable of penetrating the skin will normally penetrate certain areas of the body more efficiently than other areas. For example, penetration is very effective through the forearm and much less efficiently through the palms.

The rinsing solution, ointment, gel or cream should be thoroughly rubbed into the skin so that no excess is visible in plan view and should not be washed away until penetration through the skin has occurred, preferably for at least 15 minutes, more preferably for at least 30 minutes. Do not wash for minutes.

A known transdermal patch may be used to deliver the precursor. This is usually applied over a much longer period of time, such as 1-4 days, but generally contacts the active component over a smaller surface area, allowing slow and constant delivery of the active component.

A large number of transdermal drug delivery (drug delivery) systems have been developed and used to deliver the active compounds of the present invention. The rate of drug release in general

-54i · ** · · · · · ·.

♦·· ··* ·&lt;·. ·_{# φ}·

It is controlled by diffusion from a matrix or by passage of the active ingredient through a regulatory membrane.

Transdermal devices are well known in the mechanical field and are explained, for example, in 5,162,037, 5,154,922, 5,135,480, 4,666,441, 4,624,665, 3,742,951, 3,797,444, 4,568,343, 5,064,654, 5,071. U.S. Patent Nos. 644 and 5,071,657, the disclosure of which is incorporated herein by reference. Further explanations are provided by European Patent Application 0 279 982 and British Patent Application 2 185 187.

The device may be a known generic type comprising an adhesive matrix and a storage (reservoir) type transdermal delivery device. The device comprises fiber-based matrices containing the drug substance, wherein the fibers absorb the drug substance and / or vehicle. In the reservoir-type device, the reservoir may be bounded by a polymeric membrane that is impermeable to both the carrier and the active ingredient.

In a transdermal device, the device itself is in contact with the desired localized skin surface. In such a device, the viscosity of the active ingredient carrier is less significant than that of a cream or gel. For example, the solvent system of a transdermal device may contain oleic acid, a lactate of a straight chain alcohol and dipropylene glycol or solvent systems known per se. The active ingredient may be dissolved or suspended in the vehicle.

• ·

-55For attachment to the skin, a transdermal patch may be attached to a surgical tape with a hole pierced in the center. Preferably, the adhesive tape is covered with a removable cover which protects it before use. Generally suitable materials for this purpose are polyethylene, paper coated with polyethylene, and silicone-coated paper is preferred for easy removal. To apply the device, the protective cover is simply peeled off and the adhesive tape is attached to the patient's skin. United States Patent 5,135,480, the disclosure of which is incorporated herein by reference; other types of device may be used to attach the device to the skin without adhering accessories.

Except for the higher dose indications described above (such as contraception), the target serum concentration of DHEA is similar, regardless of whether it is used as a sex steroid precursor for the treatment of climacteria, or (alone or in combination with estrogen and / or progestin). vaginal atrophy, urinary incontinence, uterine cancer, ovarian cancer, osteoporosis, for use in the treatment of hypogonadism or decreased sex drive in accordance with the present invention; or for treating a wide variety of pathological conditions associated with reduced secretion of DHEA in the adrenal glands. It is emphasized that dosing of DHEA, DHEA-S, or any of their analogs described herein, may be correlated with the targeted serum concentration of DHEA, as all are converted directly or indirectly to DHEA via in vivo assays.

The percutaneous vein mucosal delivery system of the present invention can be used as a novel improved delivery system for the treatment of osteoporosis or other diseases that respond favorably to DHEA. To achieve these latter goals, the desired target serum concentrations are the same as those described above.

The DHEA for percutaneous or mucosal application may be in the form of the free alcohol or one or more derivatives thereof, such as valerate, benzoate, acetate, selfanthate, and (longer) alkyl ester derivatives. Dermal delivery (delivery) of DHEA or its analogs is an acceptable, easy, non-invasive way of administering such a compound. This also avoids gastrointestinal irritation, as well as the degradation of the compound and the toxicological problems attributed to the fact that DHEA first passes through the liver before reaching the general circulation.

One method of inhibiting or preventing the proliferation of breast cancer and endometrial carcinoma cells is by activating the androgen receptor with an effective compound that binds to the receptor site at low concentrations but does not significantly activate other types of steroid receptors responsible for possible side effects.

Because DHEA is a natural source of androgens [Labrie: Mole. Cell. Endocrinol. 78, C113-C118 (1991)] and this compound

-57 segregation significantly decreases with aging, and its replacement should have only minimal undesirable side effects.

Thus, the percutaneous or mucosal administration of DHEA according to the present invention provides a novel method for the prevention and therapeutic treatment of diseases that respond to the activation of the androgen receptor. Examples of such diseases are bone loss, obesity, breast cancer, endometrial cancer, ovarian cancer, urinary incontinence, hypogonadism, loss of sex drive, loss of muscle mass, loss of energy and other aging processes. The invention is also applicable to a number of diseases where estrogen receptor activation is beneficial, particularly in the treatment of osteoporosis and vaginal atrophy. The invention further provides an improved method of delivery (delivery, e.g., transdermal delivery) for diseases for which DHEA treatment is expected.

The following are examples of some preferred derivatives.

Derivatives of DHEA or DHEA-S that are expected to be transformed into DHEA or DHEA-S according to the invention under in vivo conditions may be approached by:

- Conversion of the 3B functional group to an ester which can be cleaved by an esterase (and does not produce toxic substances); and the conversion of the 17-keto group to an oxazolidine or thiazolidine moiety that is unstable in the body and *, *,,¹

II

-58 are converted to a natural precursor.

- formation of a - (acyloxy) alkyl ether in the 3fl position, which can be cleaved to an unstable semi-acetate by an esterase. The decomposition of this semi-acetal regenerates the natural precursor.

- Without modifying the 3B functional group, a

Conversion of a 17-keto group to an oxazolidine or thiazolidine moiety that is unstable in the body and converted to DHEA or DHEA-S.

The following compounds are expected to be converted in vivo to dehydroepiandrosterone (DHEA) or dehydroepiandrosterone sulfate (DHEA-S): Compounds of formula IV wherein R³represents a hydroxyl or sulfate group;

R^e is hydrogen; benzyl, aryl;

straight or branched alkyl; straight or branched alkenyl; straight or branched alkynyl; and

Z represents oxygen or sulfur.

Certain derivatives of DHEA are more lipophilic than DHEA itself, and therefore may accumulate in skin fat, and preferably release DHEA at a slow rate over a prolonged period.

In some preferred compounds of the invention, the 3-position functional group is an ester of sulfuric acid (or salts thereof) or formic acid, acetic acid, benzoic acid, butyric acid, decanoic acid, savant acid, furonic acid (furancarboxylic acid), heptanoic acid, isocaproic acid, undecanoic acid, undecylene acid, palmitic acid. propionic acid, piI ί

An ester of -59 I valine acid, propionic acid, valeric acid or carbonic acid (preferably ethyl carbonate or benzyl carbonate derivative). |

In some preferred compounds of the invention, DHEA (or DHEA-S) is modified with a 17 substituent of formula (b) or (c), wherein in formula (b) and (c)

R '·· is hydrogen; benzilesöpört; straight or branched alkyl; or straight or branched alkenyl.

The following are some of the preferred 3B ester derivatives of DHEA of formula (V).

Compounds of formula (V)

<td>The compound</td><td>X</td>

<td>Dehydroepiandrosterone 3B formate</td><td>HCO</td>

<td>3B dehydroepiandrosterone acetate</td><td>CH₃CO</td>

<td>Dehydroepiandrosterone-3B-propionate</td><td>ch₃ch₂co</td>

<td>3B dehydroepiandrosterone-butyrate</td><td>CH₃(CH₂)₂CO</td>

<td>Dehydroepiandrosterone 3B valerate</td><td>CH₃(CH₂)₃CO</td>

<td>Dehydroepiandrosterone-3B-pivalate</td><td>(CH₃)₃CCO</td>

<td>Dehydroepiandrosterone 3B benzoate</td><td>c₆h₅co</td>

<td>Dehydroepiandrosterone 3B furoate</td><td>c₄h₃OCO</td>

<td>Dehydroepiandrosterone 3B cypionate</td><td>c₅h₉(ch₂)₂co</td>

<td>Dehydroepiandrosterone 3B lactate</td><td>ch₃CHOHCOl</td>

<td>Dehydroepiandrosterone 3B decanoate</td><td>ch₃(ch₂)₈co</td>

<td>Dehydroepiandrosterone 3B undecanoate</td><td>ch₃(ch₂)₁₀co</td>

<td>Dehydroepiandrosterone 3B palmitate</td><td>CH₃(CH₂)₁₄CO</td>

<td>3B, dehydroepiandrosterone (ethyl carbonate)</td><td>c₂h₅OCO</td>

<td>3B, dehydroepiandrosterone (benzyl carbonate)</td><td>c₆h₅ch₂OCO</td>

<td>Dehydroepiandrosterone-3B-xanthine</td><td>(CH₃)₂(CH₂)₃C0</td>

<td>Dehydroepiandrosterone 3B undecylenate</td><td>H₂C = CH (CH₂)₈CO</td>

<td>3B dehydroepiandrosterone-p-enanthate</td><td>CH₃(CH₂)₅CO</td>

<td>3B, dehydroepiandrosterone (phenylpropionate)</td><td>C₆H5 (CH₂)₂CO</td>

<td>3B (Hydroxy-methoxy) -5-androsten-17-one acetate</td><td>ch₃co₂ch₂</td>

<td>3B (Hydroxy-methoxy) -5-androsten-17-one decanoate</td><td>CH₃(CH₂)₈CO₂CH₂</td>

<td colspan="3">The following table shows some of the preferred derivatives of DHEA of formula VI. Compounds of formula (VI)</td>

<td>The compound</td><td>R.. 1</td><td>Z</td>

<td>3B-hydroxy-5-androstene-17-spiro [1 ', 3'- thiazolidine-4 '- (ethyl carboxylate)]</td><td>C2H5</td><td>S</td>

<td>3B-hydroxy-5-androstene-17-spiro [1 ', 3'-thiazolidine-4' - (benzyl carboxylate)]</td><td>c₆h₅ch₂</td><td>S</td>

<td>3B-hydroxy-5-androstene-17-spiro [1 ', 3'- thiazolidine-4 '- (hexyl carboxylate)]</td><td>^c6^H13</td><td>S</td>

<td>3B-hydroxy-5-androstene-17-spiro [1 ', 3¹oxazolidine-4¹ - (ethyl carboxylate</td><td>C2 "5</td><td>SHE</td>

<td>3B-hydroxy-5-androstene-17-spiro [1 ', 3'oxazolidin-4' - (benzyl carboxylate)]</td><td>c₆h₅ch₂</td><td>SHE</td>

<td>3B-hydroxy-5-androstene-17-spiro [1 ', 3'oxazolidin-4' - (hexyl carboxylate)]</td><td>^c6^h13</td><td>SHE</td>

Corresponding 17-substituted analogs of DHEA-S may also be used.

-61ί ΐ

Production examples

First example

3β- (formyloxy) -5-androsten-17-one

Ringold et al., J. Med. Am. Chem.

Soc. 78, 816 (1956)] 2.88 g (10 mmol) of dehydroepiandroste'l are dissolved in 100 ml of 85% formic acid at 60 ° C for 1 hour. After cooling, the reaction mixture was poured into ice water, and after 16 hours, the crystalline product was filtered off and dried in vacuo. I

I

Second example

3B-acetoxy-5-androsten-17-one

Dehydroepiandrosterone (2.88 g, 10 mmol) was dissolved in 100 mL of a 1: 1 by volume mixture of acetic anhydride and pyridine and allowed to stand at room temperature for 16 hours. The mixture was then carefully poured into an ice-water mixture and after 16 hours the crystals were filtered and dried in vacuo.

Third example

Dehydroepiandrosterone 3B undecanoate

A solution of 10.2 g (50 mmol) undecanoyl chloride in 50 mL dichloromethane (DKM) was treated with 11.53 g (40 mmol) of 5-androstene-3B-ol-17-one, 14 mL (100 mmol) of triethylamine. and a solution of (dimethylamino) pyridine (0.6 g, 5 mmol) in dichloromethane (150 mL) at 5 ° C. After addition, the reaction mixture was stirred at room temperature overnight. The DKM layer

Wash with 62N water, twice with 2N hydrochloric acid, twice with 5% potassium carbonate, and brine. After removal of the solvent, the crude product was recrystallized from n-hexane / benzene to give 13.66 g (75%) of pure product, m.p. 84-85 ° C.

1 H NMR (CDCl 3)?₃): S 0.83-0.86 (m, 6H, C)₁₈CH₃ and CH₃); 1.03 (s, 3H, C₁₉CH₃); 4.58-4.61 (m, 1H, C₃-H); 5.37 (d, 1H, vinyl, J = 4.89 HZ).¹³C-NMR (CDCl₃) S1 220.90, 179.25, 139.99, 121.78, 73.38, 51.69, 50.14, 47.49, 38.12, 36.94,

36,72,

29,44,

20,31,

35,81,

29,27,

19,33,

34,68,

29,23,

14,09,

31,87,

29,09,

13,53.

31,47,

27,72,

31,42,

25,03,

30,76,

22,66,

29,53,

21,86,

4th example

36- (acyloxy) -5-androsten-17-one

The title 36-esters of dehydroepiandrosterone were prepared as follows.

A solution of dehydroepiandrosterone (mmol) in pyridine (50 mL) was added to 50 mL of acyl chloride (prepared from the appropriate acid with oxalyl chloride) in the same solvent. Thereafter, 10% (dimethylamino) pyridine was added and the reaction mixture was allowed to stand at room temperature for 16 hours. At this time, the mixture was carefully poured into ice water and extracted with ethyl acetate. The organic layer was washed successively with dilute hydrochloric acid, water, saturated sodium bicarbonate solution, water, dried and evaporated to dryness to give the ester.

il ............

5th example

3B (benzyloxy-carbonyloxy) -5-androsten-17-one

To a solution of dehydroepiandrosterone (2.88 g, 10 mmol) in DKM (100 mL) was added dropwise benzyl chloroformate (30 min) with stirring according to a known procedure [F. Reber and T. Reichstein, Helv. Chim. Acta 28, 1164 (1945)]. After stirring for 3 hours, the reaction mixture was washed with water and evaporated to dryness. The residue was dissolved in acetone and precipitated with ice-water. After 16 hours, the crystalline product is filtered off and dried in vacuo.

6th example

3B (Ethoxy-oxy-carbonyloxy) -5-androsten-17-one

The procedure described in Example 5 was followed except that ethyl chloroformate was used in place of benzyl chloroformate.

7th example

3B-hydroxy-5-androstene-17-spiro-2 '- [1¹, 3'-thiazolidine-4 '- (ethyl carboxylate)]

Following the procedure of Djerassi et al. Org. Chem. 27, 1112 (1962)], 2.88 g (10 mmol) of dehydroepiandrosterone are dissolved in anhydrous ethanol, sodium acetate is added, followed by 18 g (100 mmol) of L-cysteine ​​ethyl ester hydrochloride and the reaction mixture is heated under argon for the next day. . At this time, the reaction mixture was concentrated in vacuo and DKM was added to the residue to precipitate excess L-cysteine ​​ethyl ester hydrochloride. Subsequently, the solution was injected with il

After filtration, the filtrate was washed twice with water, dried over anhydrous magnesium sulfate and, after filtration, concentrated in vacuo. Trituration of the residue with ethanol gave a crystalline product.

8th example

3B-Hydroxy-5-androstene-17-spiro-2 - [1 ', 3 ¹ thiazolidine-4 '- (benzyl carboxylate)]

The procedure described in Example 7 was followed except that L-cysteine ​​benzyl ester hydrochloride was used instead of L-cysteine ​​ethyl ester hydrochloride.

9th example

3B-Hydroxy-5-androstene-17-spiro-2 '- [1', 3'-thiazolidine-4 '- (alkylcarboxylate)]

The procedure described in Example 7 was followed except that the hydrochloride of various L-cysteine ​​alkyl esters (e.g. L-cysteine ​​hexyl ester hydrochloride) was used instead of L-cysteine ​​ethyl ester hydrochloride.

10th example

3B-hydroxy-5-androstene-17-spiro-2 '- [1', 3'-oxazolidine-4 '- (ethyl carboxylate)]

The procedure described in Example 7 is followed except that the oxazolidine derivative is prepared using serine ethyl ester hydrochloride instead of L-cysteine ​​ethyl ester hydrochloride.

11th example

3B-Hydroxy-5-androstene-17-spiro-2 '- [1', 3'-oxazolidin-4 '- (benzylcarboxylate)]

The procedure of Example 10 is followed except that serine benzyl ester hydrochloride is used instead of serine ethyl ester hydrochloride.

12th example

3B-Hydroxy-5-androstene-17-spiro-2 '- [1', 3'-oxazolidin-4 '- (alkylcarboxylate)]

The procedure described in Example 10 was followed except that the hydrochloride of other serine alkyl esters (e.g., serine hexyl ester hydrochloride) was used instead of serine ethyl ester hydrochloride.

13th example

3B (Hydroxy-methoxy) -5-androsten-17-one acetate

To a solution of dehydroepiandrosterone (2.88 g, 10 mmol) in tetrahydrofuran (THF) (100 mL) was added sodium hydride (11 mmol, 60% in oil) under argon at room temperature. After complete reaction of the sodium hydride, chloromethyl acetate (prepared from acetyl chloride and formaldehyde or a derivative thereof using zinc chloride catalyst) is added and the reaction mixture is heated for several hours. After cooling, the mixture was poured into water and extracted with ethyl acetate. The organic phase is washed with water, dried, filtered and evaporated to dryness. This gives the desired compound.

yl

-66Examples of pharmaceutical preparations

In one aspect of the invention, DHEA (or analogues thereof) is applied to the skin or other epithelial tissue for an appropriate period of time to permit, if desired, proper penetration of the compound to produce systemic or local effects. The composition may be formulated, for example, in the form of a gel, cream, ointment or lotion, or may be applied using the delivery system disclosed in U.S. Patent Nos. 3,742,951, 3,797,494, or 4,568,343. Devices described in U.S. Patent Nos. 5,064,464, 5,071,644, or 5,071,657 may also be used to facilitate the absorption of the steroid.

Each pharmaceutical composition of the invention may contain suitable preservatives known per se.

The following non-limiting examples illustrate the preparation of a general type of cream, lotion, gel, and ointment. In addition to these carriers, one of ordinary skill in the art may select other vehicles that meet the special needs of dermatology.

14th example

A general type rinse solution contains 10% by weight of DHEA, 15% by weight of propylene glycol, 70% by weight of ethanol and 5% by weight of water.

yl

15th example

A generic gel is 10% by weight DHEA, 5% by weight propylene glycol, 0.2% by weight Carbomer 940 (available from BF Goodrich Carbopol 940^R ), 40% water, 0.2% triethylamine, 2% PPG-12-Buteh-16 (available from Union Carbide, Ucon^R fluid 50) containing 1% by weight hydroxypropyl and 41.6% by weight ethanol (in 95% ethanol containing 5% water).

16th example

A general type ointment contains 10% by weight of DHEA, 13% by weight of propylene glycol, 74% by weight of petroleum jelly, 2.9% by weight of glyceryl monostearate and 0.1% by weight of propylparaben.

17th example

A generic cream of 10% by weight DHEA, 0.2% by weight propylparaben, 5% by weight lanolin oil, 7.5% by weight sesame oil, 5% by weight cetyl alcohol, 2% by weight glyceryl monostearate, 1% by weight triethanol 5% propylene glycol, 0.1% Carbomer 940^R product and 64.2% water.

14-17 above. For each of the formulations described in Examples 1 to 4, progestin and / or estrogen may be added. For example, 0.005-0.02% 17B-estradiol and / or 0.2-2.0% medroxyprogesterone acetate may be added to the formulations with appropriate reductions in water, ethanol or petroleum jelly. The permeability of DHEA can be increased by various methods

-68 to reduce the dose used. Methods and compositions for enhancing the permeability of active compounds are disclosed in, for example, 5,051,260, 4,006,218, 3,551,554,

472 931, 4,568,343, 3,989,816 and 4,405,616.

Although the invention has been described in specific embodiments thereof, many other variations, modifications and further applications will be apparent to those skilled in the art. Advantageously, the invention is therefore limited only by the appended claims and not by the specific disclosures described.