HK1135111A - Compositions and methods for suppressing endometrial proliferation - Google Patents

Description

Compositions and methods for inhibiting endometrial hyperplasia

Technical Field

The present invention relates to compositions and methods for inhibiting endometrial hyperplasia. More specifically, the present invention relates to compositions comprising one or more progesterone antagonists for inhibiting endometrial proliferation.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims priority from the following patent applications: u.s. provisional patent application No. 60/862,632 filed 24/10/2006 and u.s. provisional patent application No. 60/885,348 filed 17/1/2007, each of which is incorporated herein by reference in its entirety.

Background

Estrogens are a group of hormones essential for a variety of physiological processes including the development of the uterus and breast, the maintenance of bone density, and cardiovascular protection through its positive effects on lipid status. The effects of estrogen are mediated through its binding to estrogen receptors in the nucleus. According to the classical model, after binding estrogen, the unoccupied estrogen receptor needs to be able to interact with DNA sequences within the promoter of the estrogen responsive gene. The DNA-bound estrogen receptor positively or negatively regulates the transcription of these genes.

Estrogens are known to have a high proliferative effect on breast and uterine tissue. For example, administration of non-antagonistic (unopposed) estrogens in menopausal women has been shown to result in endometrial hyperplasia and endometrial cancer. In contrast, progesterone is effective against estrogen-dependent endometrial hyperplasia and cancer development. Therefore, in order to combat the effects of non-antagonistic estrogens, progestins have become part of the clinical Hormone Replacement Therapy (HRT). However, recently, large clinical trials from Women's Health Initiative have shown that conjugated estrogens and medroxyprogesterone acetate increase the risk of developing cardiovascular disease, stroke, pulmonary embolism, and breast cancer. Furthermore, experimental data in surgically menopausal macaques indicate that combination therapy of estrogen and progesterone results in higher levels of breast proliferation and hyperplasia than estrogen alone. The co-administration of progesterone has been shown to be associated with intermittent bleeding, which further limits its applicability as an active agent against the highly proliferative effects of estrogens.

A number of compounds are known in the art to affect estrogen-dependent activation of estrogen receptors. Depending on a number of factors, these compounds may be fully estrogenic in that they mimic estrogen, fully antiestrogenic in that they block the action of estrogen, or they may be somewhere in between. Compounds that exhibit mixed estrogenic and antiestrogenic properties are called Selective Estrogen Receptor Modulators (SERMs). SERMs exert their estrogenic or antiestrogenic effects in a tissue-specific manner. The mechanism of this tissue specificity is not clear, but may in particular involve the recruitment of co-repressed and co-activated proteins, the relative expression levels of which may vary between tissue types, and the tissue-specific expression of the estrogen receptor isoforms α and β. Estrogen receptor α is an activator, while estrogen receptor β can inhibit estrogen receptor α activity by forming a heterodimer with the inhibitory estrogen receptor α.

The dual activity of SERMs offers several possible advantages in women. The estrogenic properties of SERMs are useful in treating or preventing diseases caused by estrogen deficiency, such as osteoporosis, while mimicking certain undesirable effects of estrogen. In contrast, the antiestrogenic effects of SERMs can be used to prevent or treat diseases such as breast cancer, where estrogenic activity is undesirable. Nonetheless, endometrial hyperplasia has been shown to be associated with SERM treatment, which limits its use.

For example, SERM tamoxifen has been shown to have an antiestrogenic effect in the breast, tamoxifen blocks the proliferative effect of estrogen in the breast, and has then been found to be beneficial in the treatment of some types of breast cancer. On the other hand, tamoxifen exhibits estrogenic effects on bone and uterus and is associated with increased incidence of endometrial hyperplasia and endometrial cancer, which limits its use as an antiestrogen.

Preliminary experiments in primates seem to indicate that antiprogestinic substances have an antiproliferative effect on the endometrium. However, there is a fear that long-term treatment with antiprogestinic substances may cause endometrial hyperplasia due to the action of non-antagonistic estrogens. Several recent studies in adult women have shown that endometrial thickening occurs during long term treatment with high doses of antiprogestin substances, which, assuming this is a result of non-antagonistic estrogenic activity, does not appear to occur at low doses.

There remains a need for a treatment regimen that is suitable for long-term administration that counteracts the proliferative effects of estrogen while maintaining the beneficial effects of estrogen on the body.

Summary of The Invention

The present invention provides a method of inhibiting endometrial hyperplasia comprising administering to a patient in need thereof an amount of a progesterone antagonist effective to inhibit endometrial tissue. The patient in need thereof may be a female suffering from endometriosis. The progesterone antagonist may be a pure antiprogestin substance or a Selective Progesterone Receptor Modulator (SPRM). In preferred embodiments, the progesterone antagonist has low affinity for the glucocorticoid receptor. In another preferred embodiment, administration of the progesterone antagonist to a female reduces luteal phase progesterone levels in the female. In another preferred embodiment, administering the progesterone antagonist to a woman does not significantly reduce estrogen levels in the woman.

In another aspect, the invention provides methods of using progesterone antagonists in estrogen and SERM therapy to prevent estrogen-dependent disorders. More specifically, the invention employs progesterone antagonists in amounts sufficient to inhibit endometrial hyperplasia in women undergoing a treatment regimen that includes administration of an estrogen or a SERM. For example, the patient may be a postmenopausal female undergoing hormone replacement therapy. In another aspect, the invention provides methods of preventing endometrial hyperplasia and/or endometrial cancer development using progesterone antagonists in the treatment of estrogens and SERMs. The progesterone antagonist can be an antiprogestin substance or a Selective Progesterone Receptor Modulator (SPRM), provided that the progesterone antagonist is used in an amount effective to inhibit endometrial hyperplasia.

In another aspect, the invention provides methods of treating pain associated with endometriosis using progesterone antagonists. Progesterone antagonists may be useful for the long-term treatment of pain associated with endometriosis.

Brief Description of Drawings

Figure 1 depicts the effect of selective progesterone receptor modulators on rat serum cortisol.

FIG. 2 depicts the dose-dependent effect of CDB-4124 on rat serum cortisol.

Detailed Description

The term "effective amount" refers to an amount of the active ingredient of the composition sufficient to achieve the desired effect, which may be, for example, inhibiting endometrial hyperplasia or treating pain associated with endometriosis.

The term "selective progesterone receptor modulator" refers to a compound that affects progesterone receptor function in a tissue specific manner. Such compounds act as progesterone receptor antagonists in certain tissues (e.g., in the uterus) and as progesterone receptor agonists in other tissues.

The term "treatment" refers to both therapeutic as well as prophylactic or preventative measures, wherein the object is to prevent or slow down (lessen) an undesired physiological change or disorder. For purposes of the present invention, beneficial or desired clinical results include, but are not limited to, alleviation of symptoms, diminishment of extent of disease, stabilization (i.e., not worsening) of the disease state, delay or slowing of disease progression, amelioration or palliation of the disease state, and remission (whether partial or complete), detectable or undetectable. "treatment" may also refer to prolonging survival as compared to that expected without receiving treatment. Individuals in need of treatment include individuals already having the condition or disease, as well as individuals predisposed to having the condition or disease, or individuals in whom prevention of the condition or disease is desired.

The term "progesterone agonist" refers to a compound that binds to the progesterone receptor and mimics the action of the natural hormone.

The term "progesterone antagonist" refers to a compound that binds to progesterone receptors and inhibits the action of progesterone.

The term "inhibit" as used herein with respect to proliferation of endometrial tissue means that mitotic proliferation of endometrial tissue is inhibited by administration of a progesterone antagonist relative to untreated endometrial tissue under the same conditions and is distinguished from cell death via, for example, apoptosis. Inhibition of progesterone antagonists in endometrial mitotic proliferation can be determined in uterine cell lines, for example, by comparing incorporation of bromodeoxyuridine (BrdU) in cells treated with progesterone antagonist to control (untreated) cells.

The term "not significantly reduced" as used herein with respect to female hormone levels means that the hormone levels remain within the normal range during administration of the compositions of the present invention. Thus, some reduction in hormone levels may occur as long as the hormone levels remain within the normal range.

The term "not significantly increased" as used herein with respect to female hormone levels means that hormone levels are maintained within normal ranges during administration of the compositions of the invention. Thus, some increase in hormone levels may occur as long as the hormone levels remain within the normal range.

The present invention relates to the use of progesterone antagonists in amounts effective to inhibit endometrial hyperplasia. These approaches stem from the unexpected discovery that some progesterone antagonists exhibit an inverse dose-dependent effect on endometrial hyperplasia while maintaining estrogen levels within the normal range. In particular, it has been found that chronic administration of high doses of antiprogestin substance/SPRMCDB-4124 inhibits endometrial hyperplasia, whereas lower doses do not inhibit endometrial hyperplasia and even tend to promote endometrial hyperplasia, although similar levels of estrogen are observed at high and low doses. This is particularly surprising in view of the following: as demonstrated below, antiprogestin substance/SPRM RU486 did not inhibit endometrial proliferation at high doses, and there have been several recent reports that chronic administration of high doses of antiprogestin substances promoted endometrial hyperplasia, presumably due to the action of non-antagonistic estrogens. The present invention also demonstrates that progesterone antagonists can be unexpectedly used to treat pain associated with endometriosis.

Endometrial hyperplasia inhibition demonstrates the use of such progesterone antagonists, as described below, during a treatment period that includes the administration of an effective amount of progesterone antagonist daily for a period of more than 6 months, where chronic/long term administration is desired. In this regard, the methods of the invention can comprise administering a composition comprising a progesterone antagonist in an amount sufficient to inhibit endometrial proliferation for a period of at least 1, 2, 3,4, 5,6, 7, 8,9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 or more days. The composition may also be administered over a dosing period of at least 1, 2, 3,4, 5,6, 7, 8,9, 10, 11, 12 or more months. The compositions may also be administered over a dosing period of at least 1, 2, 3,4, 5,6, 7, 8,9, 10 or more years. The composition may be administered daily or periodically, e.g., every other day, every other month, etc., during the dosing period. The composition may also be administered intermittently. For example, the composition is then administered for a dosing period of 1, 2, 3,4, 5 or more months, followed by a drug withdrawal period, followed by a dosing period of 1, 2, 3,4, 5 or more months, and so forth.

By "intermittent administration" is meant administration of a therapeutically effective dose of the progesterone antagonist for a period of administration, followed by a period of discontinuation, followed by another period of administration, and so forth.

By "off-period" is meant that administration of the progesterone antagonist is stopped once a day, once a week, once a month, or at a time therebetween. The period of rest may be longer or shorter than the administration period, but always longer than the administration interval during the administration period. For example, when the dosing period includes once daily, once weekly, or once monthly dosing, the rest period is at least 2 days, at least 8 days, or at least 32 days, respectively. Thus, the drug withdrawal period can be at least about 2, 3,4, 5,6, 7, 8,9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32 or more days.

In one embodiment, the compositions of the present invention are administered intermittently such that the subject experiences menstruation during at least one discontinuation period. It is expected that this method avoids the adverse effects associated with endometrial thickening or stasis which may be associated with prolonged treatment with progesterone antagonists, such as spotting, intermittent bleeding, endometrial hyperplasia or endometrial cancer. At least one, and preferably each, withdrawal period is of sufficient length to allow the individual to undergo menstruation. More preferably, the individual experiences menstruation during each discontinuance period. In a particularly preferred embodiment, the composition is administered once daily for a dosing period of 4 months, followed by a rest period during which the individual experiences menstruation, followed by another dosing period of 4 months, and so forth.

Optionally, a gonadotropin releasing hormone (GnRH) agonist or antagonist may be administered during the drug withdrawal period to accelerate endometrial shedding and regeneration. Non-limiting examples of GnRH agonists include nafarelin, buserelin, leuprorelin, triptorelin, goserelin, [ DLys [ ]⁶]GnRH、[DAla⁶]GnRH, and the like. Non-limiting examples of GnRH antagonists include histrelin, abarelix, and those disclosed in u.s. patents 4,409,208, 4,547,370, 4,565,804, 4,569,927, and 4,619,914, which are incorporated herein by reference in their entirety.

Optionally, a progestin can be administered during the withdrawal period to achieve normal menses in the patient. Administration of a progestin preferably results in a progesterone profile that mimics the natural increase and decrease of progesterone during menstruation. Such treatment regimens are well known to those skilled in the art. The administration of progestin during the withdrawal period can also provide antiestrogenic effects in addition to those achieved by administration of progesterone antagonists and thus can be useful in the treatment of estrogen-dependent conditions such as endometrial thickening. Non-limiting examples of progestins include medroxyprogesterone, megestrol, norethindrone, progesterone, hydroxyprogesterone, acetoxypregnenolone, allylestrenol, cyproterone, desogestrel, dimethisterone, ethisterone diacetate, getadene, lineestrol, and the like.

In one embodiment, a composition comprising a progesterone antagonist in an amount effective to inhibit endometrial proliferation is administered to a female patient having endometriosis. In related embodiments, a composition comprising a progesterone antagonist is administered to a female with endometriosis in an amount effective to treat headache associated with endometriosis. For example, administration of progesterone antagonists can reduce pain associated with endometrial damage. Pain is the most common and debilitating symptom of endometriosis and is the primary indication for both pharmaceutical and surgical treatment of the disease. Pain may be manifested as dysmenorrhea, pelvic pain, back pain, abdominal pain, breast pain, dyspareunia, and the like. Administration of progesterone antagonists can also reduce the size of endometrial lesions. Current regimens for the treatment of endometriosis include GnRH agonists, which induce a state of false menopause by inhibiting ovarian estrogen secretion and therefore cannot be used for long-term administration, as this would lead to loss of bone density, loss of calcium throughout the body and other osteoporosis-like side effects. The compositions of the present invention can be administered for extended periods without significantly reducing estrogen levels.

In another embodiment, the invention provides methods of treating estrogen-dependent conditions associated with current hormone therapy employing an estrogenic compound such as an estrogen or SERMS, comprising co-administering an effective amount of a progesterone antagonist to inhibit endometrial hyperplasia. In this regard, the progesterone antagonist can be administered before, during or after the estrogen or SERMS as part of a combined hormone treatment regimen.

In another embodiment, the invention provides a method of hormone replacement comprising administering to a menopausal or postmenopausal woman, an effective amount of a progesterone antagonist and an estrogenic compound, wherein the amount of progesterone antagonist is effective to inhibit estrogen-dependent disorders. The estrogenic compound may be an estrogen or a SERM. The estrogenic compound may be administered prior to the progesterone antagonist. Followed by administration, or concurrently with administration of the progesterone antagonist. The estrogen-dependent disorder may be, but is not limited to, endometrial hyperplasia, endometrial hyperproliferation, or endometrial cancer.

In a preferred embodiment of each of the methods of the invention, administering the progesterone antagonist to the female does not significantly reduce estrogen levels in the female. Thus, the present invention provides advantages over current treatments for the treatment of endometriosis, which often employ gonadotropin releasing hormone (GnRH) agonists such as(leuprolide acetate).

In another preferred embodiment of each of the methods of the invention, the administration of the progesterone antagonist to the female does not significantly increase progesterone levels in the female. More preferably, administration of the progesterone antagonist to a woman reduces progesterone levels, particularly luteal phase progesterone levels, in the woman.

In another preferred embodiment of each of the methods of the invention, the progesterone antagonist exhibits reduced affinity for the glucocorticoid receptor. More preferably, the binding affinity of the progesterone antagonist for the progesterone receptor is at least 1.5 times greater than the binding affinity of the progesterone antagonist for the glucocorticoid receptor.

Any known progesterone antagonist characterized by the above compounds can be used by those skilled in the art in the practice of the present invention. Particularly useful compounds include those disclosed in U.S. patent 6,900,193, which is incorporated herein by reference in its entirety, and those disclosed in U.S. patent 6,861,415, which is incorporated herein by reference in its entirety, which is a 21-substituted 19-norpregnane having the general formula:

wherein:

x may be, for example, alkyl, alkenyl, alkynyl, hydrogen, halogen, monoalkylamino or dialkylamino, such as N, N-dimethylamino;

R₁may be for example O, NOH or NO-methyl;

R₂may be, for example, hydrogen or acetyl; and is

R₃May be, for example, methoxy, formyloxy, acetoxy, acyloxy, S-alkoxy, acetylthienylmethyl (theonyl), glycinate, vinyl ether, acetoxymethyl, methylcarbonate, halogen, methyl, hydroxy and ethoxy.

Examples of 21-substituted 19-norpregnanes include, but are not limited to, the following 24 disclosed compounds.

CDB-4247 (21-propionyloxy-17 α -acetoxy-11 β - (4N, N-dimethylaminophenyl) -19-norpregnane-4, 9-diene-3, 20-dione) having the following structural formula:

CDB-4361 (21-vinyl ether-17 α -acetoxy-11 β - (4N, N-dimethylaminophenyl) -19-norpregnane-4, 9-diene-3, 20-dione) having the following structural formula:

CDB-4059 (21-acetoxy-17 α -acetoxy-11 β - (4N, N-dimethylaminophenyl) -19-norpregnane-4, 9-diene-3, 20-dione) having the following structural formula:

CDB-4124 (21-methoxy-17 α -acetoxy-11 β - (4N, N-dimethylaminophenyl) -19-norpregnane-4, 9-diene-3, 20-dione) having the following structural formula:

CDB-4031 (21-bromo-17 α -acetoxy-11 β - (4N, N-dimethylaminophenyl) -19-norpregnane-4, 9-diene-3, 20-dione) having the following structural formula:

CDB-3876 (21-chloro-17 α -acetoxy-11 β - (4N, N-dimethylaminophenyl) -19-norpregnane-4, 9-diene-3, 20-dione) having the following structural formula:

CDB-4058 (21-fluoro-17 α -acetoxy-11 β - (4N, N-dimethylaminophenyl) -19-norpregnane-4, 9-diene-3, 20-dione) having the following structural formula:

CDB-4030 (21-methyl-17 α -acetoxy-11 β - (4N, N-dimethylaminophenyl) -19-norpregnane-4, 9-diene-3, 20-dione) having the following structural formula:

CDB-4152 (21-hydroxy-17 α -acetoxy-11 β - (4N, N-dimethylaminophenyl) -19-norpregnane-4, 9-diene-3, 20-dione) having the following structural formula:

CDB-4167 (21-ethoxy-17 α -acetoxy-11 β - (4N, N-dimethylaminophenyl) -19-norpregnane-4, 9-diene-3, 20-dione) having the following structural formula:

CDB-4101 (21-methoxysulfanyl-17 α -acetoxy-11 β - (4N, N-dimethylaminophenyl) -19-norpregnane-4, 9-diene-3, 20-dione) having the following structural formula:

CDB-4110 (21-acetonide-17 α -acetoxy-11 β - (4N, N-dimethylaminophenyl) -19-norpregnane-4, 9-diene-3, 20-dione) having the following structural formula:

CDB-4111(21-BMD-17 α -acetoxy-11 β - (4N, N-dimethylaminophenyl) -19-norpregnane-4, 9-diene-3, 20-dione) having the following structural formula:

14.CDB-4125(21-(Cyp^*-hydroxy) -17 α -acetoxy-11 β - (4N, N-dimethylaminophenyl) -19-norpregnane-4, 9-diene-3, 20-dione) having the following structural formula:

^*cyp ═ 3-cyclopentylpropionyloxy-

CDB-4205 (3-hydroxyamino-21-methoxy-17 α -acetoxy-11 β - (4N, N-dimethylaminophenyl) -19-norpregnane-4, 9-diene-3, 20-dione) having the following structural formula:

CDB-4206 (3-hydroxyamino-21-acetoxy-17 α -acetoxy-11 β - (4N, N-dimethylaminophenyl) -19-norpregnane-4, 9-diene-3, 20-dione) having the following structural formula:

CDB-4226 (3-hydroxyamino-21-ethoxy-17 α -acetoxy-11 β - (4N, N-dimethylaminophenyl) -19-norpregnane-4, 9-diene-3, 20-dione) having the following structural formula:

CDB-4262 (3-methoxyamino-21-ethoxy-17 α -acetoxy-11 β - (4N, N-dimethylaminophenyl) -19-norpregnane-4, 9-diene-3, 20-dione) having the following structural formula:

CDB-4223 (21-methylsulfanyl-17 α -acetoxy-11 β - (4N, N-dimethylaminophenyl) -19-norpregnane-4, 9-diene-3, 20-dione) having the following structural formula:

CDB-4119 (4-benzoin-21-acetylthio-17 α -acetoxy-11 β - (4N, N-dimethylaminophenyl) -19-norpregnane-4, 9-diene-3, 20-dione) having the following structural formula:

CDB-4239 (4-benzoin-21-methoxy-17 α -acetoxy-11 β - (4N, N-dimethylaminophenyl) -19-norpregnane-4, 9-diene-3, 20-dione) having the following structural formula:

CDB-4306 (21-glycine ester-17 α -acetoxy-11 β - (4N, N-dimethylaminophenyl) -19-norpregnane-4, 9-diene-3, 20-dione) having the following structural formula:

CDB-4352 (21-cyanothio-17 α -acetoxy-11 β - (4N, N-dimethylaminophenyl) -19-norpregnane-4, 9-diene-3, 20-dione) having the following structural formula:

CDB-4362 (21-methoxyacetyl-17 α -acetoxy-11 β - (4N, N-dimethylaminophenyl) -19-norpregnane-4, 9-diene-3, 20-dione) having the following structural formula:

the 11 β -monodemethylated derivatives of compound 24 disclosed above (i.e., compounds wherein X is N-methylamino) are particularly useful in the practice of the present invention. In this regard, it has been demonstrated that the mono-demethylated derivative of CDB-4453 (21-methoxy-17 α -acetoxy-11 β - (4-N-methylaminophenyl) -19-norpregnane-4, 9-diene-3, 20-dione), CDB-4124, has lower anti-glucocorticoid activity than its parent compound. Attardi et al, 2002, mol.cell.endocrin.188: 111-123, the contents of which are incorporated herein by reference.

Although the compounds of the above formula and their mono-demethylated derivatives are preferred, any progesterone antagonist can be used in the practice of the present invention because it has an antagonist effect on the progesterone receptor. Preferably, the progesterone antagonist has one or more of the following characteristics: low anti-glucocorticoid activity, minimal estrogenic and anti-estrogenic activity, and no significant increase in progesterone levels.

Antiprogestinic substances that may be used in the present invention include, but are not limited to, asoprisnil (benzaldehyde, 4- [ (11 β,17 β) -17-methoxy-17- (methoxymethyl) -3-oxoestra-4, 9-dien-11-yl ] -1- (E) -oxime; J867), its metabolite J912(4- [17 β -hydroxy-17 α - (methoxymethyl) -3-oxoestra-4, 9-dien-11 β -yl ] benzaldehyde- (1E) -oxime), as well as other compounds described in DE 4332283 and DE 4332284; CDB-2914(17 α -acetoxy-11 β - (4-N, N-dimethylaminophenyl) -19-norpregnane-4, 9-diene-3, 20-dione) and Stratton et al, 2000, hu. reprod.15: 1092-1099; JNJ-1250132 and Allan et al, 2006, steroid 71: 949-954; zhi et al, 1998, j.med.chem.41: 291-302, 5-aryl-1, 2-dihydrochromeno [3, 4-f ] quinoline; 1, 4-dihydro-benzo [ d ] [1, 3] oxazin-2-one described in Zhang et al, u.s. patents 6,509,334, 6,566,358 and 6,713,478; 1, 3-dihydro-indol-2-ones described in U.S. patent 6,391,907 to fentome et al; 2, 3-dihydro-1H-indole described in U.S. patent 6,417,214 to Ulrich et al; benzimidazolone compounds and analogs thereof described in U.S. patent 6,380,235 to Zhang et al; 2, 1-benzisothiazoline 2, 2-dioxide as described in U.S. patent 6,339,098 to Collins et al; cyclic aminocaproate esters and cyclic amide compounds described in U.S. patents 6,306,851 and 6,441,019 to Santilli et al; cyclic urea and cyclic amide derivatives described in U.S. patent 6,369,056 to Zhang et al; and quinazolinone and benzoxazine derivatives as described in Zhang et al, U.S. patent 6,358,948.

Other antiprogestinic substances useful in the present invention include, but are not limited to, (6 α, 11 β,17 β) -11- (4-dimethylaminophenyl) -6-methyl-4 ', 5' -dihydrospiro [ estra-4, 9-diene-17, 2 '(3' H) -furan ] -3-one (ORG-31710) and other compounds described in us patent 4,871,724; (11 β,17 α) -11- (4-acetylphenyl) -17, 23-epoxy-19, 24-dicarballyl-4, 9, 20-trien-3-one (ORG-33628); (7 β, 11 β,17 β) -11- (4-dimethylaminophenyl-7-methyl ] -4 ', 5' -dihydrospiro [ estra-4, 9-diene-17, 2 '(3' H) -furan ] -3-one (ORG-31806) and other compounds described in U.S. Pat. No. 4,921,845, ZK-112993 and other compounds described in Michna et al, 1992, J.Steroid biochem, Molec.biol.41: 339-348; ORG-31376; ORG-33245; ORG-31167; ORG-31343; RU-2992; RU-1479; RU-25056; RU-49295; RU-46556; RU-26819; 1127; 120753; LG 120830; LG 1447; LG 1216; CGP-19984A; RTI-3021-012; RTI-3021-022; RTK-35022; Z9826-K-3526; ZJ-35022-136796; ZJ-355631; Z9826; ZJ-365631; Z9826) (ii) a ZK-98734; and ZK-137316.

Other antiprogestinic substances useful in the present invention include, but are not limited to, mifepristone (11 β - [ p- (dimethylamino) phenyl ] -17 β -hydroxy-17- (1-propynyl) estra-4, 9-dien-3-one; RU 486) and other compounds described in U.S. patents 4,386,085, 4,447,424, 4,519,946 and 4,634,695; jiang et al, 2006, steroid 71: 949-954; onapristone (11 β - [ p- (dimethylamino) phenyl ] -17 α -hydroxy-17- (3-hydroxypropyl) -13 α -estra-4, 9-dien-3-one) and other compounds described in U.S. patent 4,780,461; rilosone (((Z) -11 β - [ (4-dimethylamino) phenyl ] -17- β -hydroxy-17 α - (3-hydroxy-1-propenyl) estra-4, 9-dien-3-one) and other compounds described in U.S. Pat. No. 4,609,651, Belagner et al, 1981, steroids 37: 361-382, 11 β -substituted 19-nor steroids such as 11 β - (4-methoxyphenyl) -17 β -hydroxy-17 α -ethynyl-4, 9-estradien-3-one, and 11 β -aryl-4-estrene compounds such as (Z) -11 β - [ (4-dimethylamino) phenyl) ] -17 β -hydroxy-17 α - (3-hydroxy-3-dien-3-one described in U.S. Pat. No. 5,728,689 -1-propenyl) estr-4-en-3-one; 11 β -aryl-estrene derivatives described in U.S. patents 5,843,933 and 5,843,931; 11-benzaldoxime-estra-diene derivatives described in U.S. patent 5,693,628 such as 4- [17 β -methoxy-17 α - (methoxymethyl) -3-oxoestra-4, 9-dien-11 β -yl ] benzaldehyde-1- (E) -oxime; 11-benzaldoxime-17 β -methoxy-17 α -methoxymethyl-estradiene derivatives described in U.S. patent 5,576,310 such as 4- [17 β -methoxy-17 α - (methoxymethyl) -3-oxoestra-4, 9-dien-11 β -yl ] benzaldehyde-1- (E) - [ O- (ethylamino) carbonyl ] oxime; s-substituted 11 β -benzaldoxime-estra-4, 9-diene-carbonic acid thiol esters described in WO 99/45023, for example 4- [17 β -methoxy-17 α - (methoxymethyl) -3-oxoestra-4, 9-dien-11 β -yl ] benzaldehyde-1- (E) - [ O- (ethylthio) carbonyl ] oxime; steroid esters such as (Z) -6 ' - (4-cyanophenyl) -9, 11 α -dihydro-17 β -hydroxy-17 α - [4- (1-oxo-3-methylbutoxy) -1-butenyl ]4 ' H-naphtho [3 ', 2 ', 1 '; 10, 9, 11] estr-4-en-3-one; fluoro 17 α -alkyl chain steroids described in WO98/34947 such as 11 β - (4-acetylphenyl) -17 β -hydroxy-17 α - (1, 1, 2, 2, 2-pentafluoroethyl) estra-4, 9-dien-3-one; 17-spirofuran-3' -ylidene steroids described in U.S. patent 5,29,878 such as 11 β - (4-acetylphenyl) -19, 24-dinor-17, 23-epoxy-17 α -chol-4, 9, 20-trien-3-one; (Z) -11 β, 19- [4- (3-pyridyl) -o-phenylene ] -17 β -hydroxy-17 α - [ 3-hydroxy-1-propenyl ] -4-androsten-3-one and other compounds described in U.S. patent 5,439,913; 13-alkyl-11-beta-phenyl stanol compounds described in U.S. patent 5,446,036 such as 11 beta- [4- (1-methylvinyl) phenyl ] -17 alpha-hydroxy-17 beta- (3-hydroxypropyl) -13 alpha-estra-4, 9-dien-3-one; 11-aryl steroids described in U.S. patent 4,921,845 such as 4 ', 5' -dihydro-11 β - [4- (dimethylamino) phenyl ] -6 β -methylspiro [ estra-4, 9-diene-17 β,2 '(3' H) -furan ] -3-one; 11- β -aryl-estradiene compounds described in U.S. patents 4,829,060, 4,814,327 and 5,089,488; 11- β -aryl-4, 9-steradiene compounds and 11- β -aryl-13-alkyl-4, 9-steradiene compounds described in U.S. patents 5,739,125, 5,407,928 and 5,273,971; 11- β -aryl-6-alkyl (or alkenyl or alkynyl) steroids described in EP 289073; 10- β, 11- β -bridged steroids described in U.S. patent 5,093,507; 11- β -aryl-14- β -steroids described in U.S. patent 5,244,886; 19, 11- β -bridged steroids described in U.S. patents 5,095,129, 5,446,178, 5,478,956 and 5,232,915; 1-arylsulfonyl, arylcarbonyl, and 1-arylphosphono-3-phenyl-1, 4,5, 6-tetrahydropyridazine compounds described in U.S. patent 5,684,151; 1-arylsulfonyl, arylcarbonyl, and arylthiocarbonylpyridazino derivatives described in U.S. patent 5,753,655; 1, 2-dihydro- [1, 2-g ] quinoline derivatives and 1, 2-dihydro-chromeno [3, 4-f ] quinoline derivatives described in U.S. patents 5,688,808, 5,693,646, 5,693,647, 5,696,127, 5,696,130 and 5,696,133; kang et al, 2007, bioorg.med.chem.lett.15: 907-; and Kang et al, 2007, bioorg.med.chem.lett.17: 2531-2534, and 7-oxa-steroid 4.

In a preferred embodiment, the progesterone antagonist is antiprogestin/SPRMCDB-4124 (21-methoxy-17 α -acetoxy-11 β - (4N, N-dimethylaminophenyl) -19-norpregnane-4, 9-diene-3, 20-dione). Example 10 demonstrates that administration of CDB-4124 at high doses (50 mg/day) inhibits endometrial hyperplasia in adult females, but not at lower doses (25 mg/day and 12.5 mg/day).

The progesterone antagonist compositions of the invention can be administered to a patient undergoing any hormonal treatment associated with high risk or incidence of endometrial hyperplasia or endometrial cancer. Such treatments may include, but are not limited to, administration of estrogen or administration of a SERM. The progesterone antagonist compositions of the present invention can also be administered to a patient undergoing anti-estrogen therapy because the patient can benefit from the antiproliferative effect exerted by the progesterone antagonist compounds in endometrial tissue of the uterus.

SERMs are currently administered to treat a variety of different disorders, including breast cancer, osteoporosis, colon cancer, neurodegenerative diseases such as parkinson's disease and alzheimer's disease, cardiovascular disease, vaginal atrophy and obesity. However, SERM treatment is associated with endometrial hyperplasia and endometrial cancer. For example, tamoxifen treatment of breast cancer results in approximately 20% of women with intact uterus with atypical hyperplasia incidence. Patients with endometrial specimens exhibiting atypical have a 25% probability of developing cancer. The compounds of the invention are administered in a dosage sufficient to combat the proliferation associated with SERM treatment. The compounds of the invention may be administered in combination with a SERM to treat any of the above conditions.

Estrogens are currently administered to postmenopausal women who are no longer producing estrogen as part of Hormone Replacement Therapy (HRT). However, for menopausal women with an intact uterus, estrogen therapy alone is considered unsafe because of the concomitant endometrial hyperplasia. The antiproliferative effect of anti-estrogens is achieved by frequently and clinically co-administering progestogens; however, in the WHI study, the addition of progestagen has been shown to be associated with breast cancer and can lead to intermittent bleeding. The compounds of the present invention may be administered in combination with an estrogen as part of a hormone replacement therapy.

The disclosed compounds may act as progesterone antagonists in the uterus. The compounds of the present invention may be suitable for long term use in the treatment of other indications in menopausal patients undergoing hormone replacement therapy. When such applications are considered, the compounds of the present invention preferably have low glucocorticoid receptor binding activity, and therefore, the compounds of the present invention do not significantly interfere with the function of the glucocorticoid receptor. Administration of the compounds of the invention may have reduced side effects such as mood swings, fatigue and weight loss, which are typically found when antiprogestins with high affinity for the glucocorticoid receptor are used.

In another embodiment, the invention provides methods useful for identifying compounds having selective progesterone receptor binding activity. These methods include receptor binding and in vivo bioassays such as anti-McGinty, anti-Clauberg, glucocorticoid, estrogen, androgen, anti-glucocorticoid (AG), anti-estrogen and anti-androgen activity and post-coital and anti-ovulation activities, using the compounds of the present invention as a reference.

In another embodiment, the present invention teaches that the effect of potential SPRMs on transcriptional activity in human cells can also be analyzed. When using SPRMs disclosed in the present invention as a reference, the analysis may provide information on: (1) SPRMs interact with receptors, (2) activated receptors interact with other transcription factors, (3) activating transcription complexes on Progesterone Response Elements (PREs), and ultimately their effect on gene expression. In these experiments, a plasmid expressing hPR-B can be co-transfected into HeLa, HepG2 or T47D cells under a PRE-dependent promoter using any receptor known to those skilled in the art. Receptors may include, but are not limited to, luciferase, β -galactosidase, green fluorescent protein, red fluorescent protein, or yellow fluorescent protein. After transfection, cells are treated with the test compound or one of the SPRMs disclosed in this application as a positive control. After treatment, the cells are analyzed for reporter gene expression.

In another embodiment, the present invention teaches that the ability of the contemplated SPRMs to resist dexamethasone-induced cell death can be tested in the human lymphocyte cell line CEM-7 and compared to the effects of SPRMs disclosed in the present specification. In these experiments, dexamethasone can be added at concentrations that result in cell death. The cells are then treated with RU486, a SPRM of the invention or test compound at 10^-6And 10^-8Concentration of MAnd (5) degree processing.

Progesterone antagonist compounds useful according to the present invention can be synthesized using synthetic chemical techniques known in the art, for example, as disclosed in U.S. patent No. 6,861,415. It will be appreciated that under reaction conditions some functional groups may interact with other reactants or reagents and therefore may require temporary protection. The use of protecting Groups is described in 'Protective Groups in organic Synthesis', 2^ndEdition, t.w.greene&M. Wutz, Wiley-Interscience (1991).

In one embodiment, the compositions of the present invention comprise one or more progesterone antagonists or pharmaceutically acceptable salts thereof. Depending on the process conditions, the salt compounds can be obtained in neutral or salt form. Salt forms include hydrates and other solvates as well as crystalline polymorphs. These end products in free base as well as salt form can be used according to the invention.

Acid addition salts can be converted into the free bases in a manner known per se using basic reagents, for example bases, or by ion exchange. The obtained free base may also form a salt with an organic or inorganic acid.

In the preparation of acid addition salts, it is preferred to use acids which form suitable pharmaceutically acceptable salts. Examples of such acids are hydrochloric acid, sulfuric acid, phosphoric acid, nitric acid, aliphatic acids, cycloaliphatic carboxylic or sulfonic acids, such as formic acid, acetic acid, propionic acid, succinic acid, glycolic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, glucuronic acid, fumaric acid, maleic acid, hydroxymaleic acid, pyruvic acid, aspartic acid, glutamic acid, p-hydroxybenzoic acid, pamoic acid, ethanesulfonic acid, hydroxyethanesulfonic acid, phenylacetic acid, mandelic acid, benzenesulfonic acid (aldobensenesulfonic acid), toluenesulfonic acid, galactaric acid, galacturonic acid or naphthalenesulfonic acid. All crystalline polymorphs can be used according to the present invention.

Base addition salts may also be used in accordance with the present invention and may be prepared by: the free acid form is contacted with a sufficient amount of the desired base in a conventional manner to form a salt. The free acid form may be regenerated by contacting the salt form with an acid and isolating the free acid in a conventional manner. Pharmaceutically acceptable base addition salts are formed with metals or amines, such as alkali and alkaline earth metals or organic amines. Examples of metals used as cations are sodium, potassium, calcium, magnesium, and the like. Examples of suitable amines are amino acids such as lysine, choline, diethanolamine, ethylenediamine, N-methylglucamine and the like.

For the purposes described above, the compounds of the invention may be administered via any conventional route in which progesterone antagonists are active. For example, the progesterone antagonists of the present invention can be administered orally, parenterally, sublingually, transdermally, rectally, transmucosally, topically, via inhalation, buccally, or combinations thereof. Parenteral administration includes, but is not limited to, intravenous administration, intraarterial administration, intraperitoneal administration, subcutaneous administration, intramuscular administration, intrathecal administration, intraarticular administration, intracisternal administration, and intraventricular administration. The administration form may be tablets, capsules, pills, nasal sprays, pellets, implants (or other depot formulations) and the like.

The therapeutically effective amount of the composition required for treatment may vary depending on the following factors: the particular compound used, the mode of administration, the severity of the condition being treated, the length of time of desired activity, etc., and will ultimately be at the discretion of the attendant clinician. In all cases, an effective amount of a particular compound is an amount sufficient to inhibit endometrial hyperplasia. Generally, however, for human treatment, the dose will generally be from about 0.001mg/kg to about 500 mg/kg/day, for example from about 1 μ g/kg to about 1 mg/kg/day or from about 1 μ g/kg to about 100 μ g/kg/day. For most large mammals, the total daily dose is from about 1 to 100mg, preferably from about 2 to 80 mg. Dosage regimens may be adjusted to provide the optimum therapeutic response. The desired dose may conveniently be administered in a single dose, or as multiple doses administered at appropriate intervals, for example as 2, 3,4 or more sub-doses per day.

By way of example, the compositions of the present invention may be administered to a subject to provide the following amounts of progesterone antagonist to the subject: about 1. mu.g/kg to about 1mg/kg body weight, e.g., about 1. mu.g/kg, about 25. mu.g/kg, about 50. mu.g/kg, about 75. mu.g/kg, about 100. mu.g/kg, about 125. mu.g/kg, about 150. mu.g/kg, about 175. mu.g/kg, about 200. mu.g/kg, about 225. mu.g/kg, about 250. mu.g/kg, about 275. mu.g/kg, about 300. mu.g/kg, about 325. mu.g/kg, about 350. mu.g/kg, about 375. mu.g/kg, about 400. mu.g/kg, about 425. mu.g/kg, about 450. mu.g/kg, about 475. mu.g/kg, about 500. mu.g/kg, about 525. mu.g/kg, about 550. mu.g/kg, about 575. mu.g/kg, about 600. mu.g/kg, about 625. mu.g/kg, about 675 μ g/kg, about 700 μ g/kg, about 725 μ g/kg, about 750 μ g/kg, about 775 μ g/kg, about 800 μ g/kg, about 825 μ g/kg, about 850 μ g/kg, about 875 μ g/kg, about 900 μ g/kg, about 925 μ g/kg, about 950 μ g/kg, about 975 μ g/kg or about 1mg/kg body weight.

The compositions of the present invention may comprise from about 25 to about 90% by weight of the active ingredient, more typically from about 5% to 60% by weight of the active ingredient and a carrier.

Solid carriers may include starch, lactose, dicalcium phosphate, microcrystalline cellulose, sucrose and kaolin, while liquid carriers may include sterile water, polyethylene glycols, nonionic surfactants, and edible oils such as corn, peanut and sesame oils, as are appropriate to the nature of the active ingredient and the particular form of administration desired. Flavoring agents, coloring agents, preservatives and antioxidants such as vitamin E and ascorbic acid may also be included in the formulations. Under normal conditions of storage and use, the formulation may contain a preservative to prevent the growth of microorganisms.

The compositions of the present invention can be formed into tablets in a tablet press using techniques well known to those skilled in the art. Optionally, the active ingredients of the present invention may also be compressed individually into bilayer tablets. According to the invention, the tablet may contain an antiestrogen, an estrogen or a SERM as one of the active ingredients. The compositions of the present invention may also be formulated as oily solutions.

For patients treated with the composition of the present invention, their serum estrogen and glucocorticoid levels should be monitored regularly.

The following non-limiting examples are provided to aid in understanding the teachings of the present invention.

All patents, patent applications, and publications mentioned herein are incorporated by reference to the fullest extent allowed by law.

Example 1. the formulations of the present invention may be formulated into tablets.

To obtain tablets for use in the practice of the present invention, the following components may be compressed together in a tablet press:

50.0mg CDB-4124

140.5mg lactose

69.5mg corn starch

2.5mg of poly-N-vinylpyrrolidone

2.0mg silica aerogel

0.5mg magnesium stearate

To obtain a bilayer tablet for the practice of the present invention, the following components can be compressed together in a tablet press:

20.0mg tamoxifen

50.0mg CDB-4124

105.0mg lactose

40.0mg rice starch

2.5mg of Poly-N-vinylpyrrolidone 25

2.0mg silica aerogel

0.5mg magnesium stearate

To obtain tablets containing an antiestrogen substance for use in the practice of the present invention, for example, the following components may be compressed together in a tablet press:

10.0mg raloxifene

50.0mg CDB-4124

125.0mg lactose

50.0mg rice starch

2.5mg of Poly-N-vinylpyrrolidone 25

2.0mg silica aerogel

0.5mg magnesium stearate

To obtain an oily preparation for carrying out the invention, for example, the following components may be mixed together and added to an ampoule:

100.0mg CDB-4124

343.4mg Castor oil

608.6mg of benzyl benzoate

Example 2 the compounds of the present invention may have only weak anti-glucocorticoid receptor binding activity.

Some antiprogestins were tested for their ability to bind to rabbit progesterone receptor (rbPR) and glucocorticoid receptor (rbGR) in a receptor binding assay. Briefly, PR-or GR-containing cytoplasms from uterus or thymus, respectively, of estradiol-administered immature rabbits were prepared in TEGMD buffer (10mM Tris, pH7.2, 1.5mM EDTA, 0.2mM sodium molybdate, 10% glycerol, 1mM DTT). For PR binding, the cytoplasm is bound to 6nM 1, 2-, [ 2 ]³H]Progesterone (50.0Ci/mmol) was incubated together and competitor was added at a concentration of 2-100 nM. For GR binding, the cytoplasm is bound to 6nM 6, 7-, [ 2 ]³H]Dexamethasone (40Ci/mmol) was incubated together and competitor was added at a concentration of 2-100 nM. After overnight incubation at 4 ℃, bound and unbound [ 2 ] were separated by adding dextran-coated charcoal and centrifuging at 2100 Xg for 15 minutes at 4 ℃, [ 2 ]³H]A steroid compound. Will contain³H]The supernatant of the steroid receptor complex was decanted to a volume containing 4ml Optifluor (Packard Instrument C)o.) vial, vortexed, equilibrated in a liquid scintillation counter for 30 minutes, and then counted for 2 minutes. By inputting count data into a four-parameter sigmoidal computer program (Immunoassay Data reduction program, Packard Instrument Co., Meriden.) to determine the EC for each standard curve and for each compound curve₅₀(effective concentration). The Relative Binding Affinity (RBA) of each compound was calculated using the following formula: EC of Standard₅₀EC of test Compound₅₀X 100. The standards for PR and GR assays were unlabeled progesterone and dexamethasone, respectively. The results of these experiments are summarized in table 1, expressed as the ratio of the relative binding affinities of each compound for the rbPR and rbGR receptors (rbPR/rbGR). This difference reflects the relative activity of the compounds in cells or tissues bearing these two receptors and the necessary proprietary cofactors.

The relative biological activity of the same compounds in the rabbit uterus as tested by the anti-McGinty and anti-Clauberg assays is also given in table 1. For these experiments, the compound CDB-2914 (listed at the end of the table) was used as a control or reference compound (rabbit bioactivity ═ 1.00) since the results of experiments using CDB-2914 have been published (Hild-Petito et al, 1996; Passaro et al, 1997; Reel et al, 1998; Larner et al, 2000). For the anti-McGinty test, immature female rabbits received a subcutaneous injection of 5 μ g estradiol in 10% ethanol/sesame oil once a day for 6 consecutive days. On day 7, animals were subjected to sterile abdominal surgery to ligate two segments of 3-4cm of uterine horn. Test compounds in an appropriate solvent are injected intraluminally into the ligation segment of one uterine horn and a separate carrier is intraluminally injected into the ligation segment of another uterine horn. For the next three days, a stimulating dose of progesterone (267 μ g/day) was injected daily subcutaneously into each rabbit to induce endometrial hyperplasia. All animals were sacrificed on day 10 to remove the uterus, where the central segment of the ligature was removed and fixed in 10% neutral buffered formalin and histologically processed. The extent of endometrial gland hyperplasia was evaluated microscopically for 5 microtomes stained with hematoxylin and eosin. The percent inhibition of endometrial hyperplasia was calculated for each rabbit and the mean value was recorded for the group comprising 5 rabbits. For the anti-Clauberg test, immature rabbits received subcutaneous injections of 5 μ g estradiol in 10% ethanol/sesame oil for 6 consecutive days. On day 7, animals received progesterone by subcutaneous injection (160 μ g/day) and experimental compound in the appropriate vehicle either orally or subcutaneously for 5 consecutive days. One group of rabbits received progesterone only. 24 hours after the last dose, all animals were sacrificed to remove the uterus, washed of all fat and connective tissue from the uterus, weighed to the nearest 0.2mg, and placed in 10% neutral buffered formalin for subsequent histological processing. The extent of endometrial gland hyperplasia was evaluated microscopically for 5 microtomes stained with hematoxylin and eosin. The percent inhibition of endometrial hyperplasia of the test compounds at each dose level was obtained by comparison to animals stimulated with progesterone alone. The data presented in table 1 (rabbit bioactivity) reflect the average results obtained for each compound relative to CDB-2914 by anti-McGinty and anti-Clauberg analysis.

The antiprogestin substances tested were ranked based on the selectivity of each compound for rabbit PR versus rabbit GR, as listed in table 1. Antiprogestin substances have also been graded based on the biological activity of the uterus of rabbits. The data presented in table 1 indicate that the affinity of the main compound for the progesterone receptor is at least 1.5 times its affinity for the glucocorticoid receptor.

The results of these experiments also show that the two main compounds CDB-4124 and CDB-4059 have stronger antiprogestinic activity in rabbit uterus than RU486 and CDB-2914. Both compounds lack estrogenic, androgenic, estrogenic and antiandrogenic activity. Both compounds have minimal anti-glucocorticoid receptor activity, a feature that distinguishes them from RU486 and CDB-2914, which have moderate activity in glucocorticoid receptor binding. In these assays, CDB-4124 performed slightly better than CDB-4059.

TABLE 1 receptor binding and biological Activity of SPRMS

EXAMPLE 3 Cortisol was determined.

Several different experimental systems support the conclusion that RU486 increases cortisol because RU486 has strong antiglucocorticoid properties in humans and primates.

However, as shown in figure 1, rats treated with RU486 at 10mg/kg showed no significant difference in cortisol levels. In contrast, rats treated with CDB-4124 or CDB-4059 at the same dose level had significantly higher serum cortisol levels than the control group.

These higher levels are in the range of 3-4ug/dl (30-40 ng/ml). These effects were dose-dependent, as an increase in the dose of CDB-4124 resulted in an increase in cortisol (figure 2).

This difference in the effect of RU486 on cortisol levels relative to CDB-4124 or CDB-4059 could be explained by the assumption that rat liver was able to metabolize RU486 better than both CDB compounds after a long-term administration of 21 days.

Example 4. Cortisone was determined.

Corticosterone is the largest glucocorticoid content in rats. The effect of SPRMs on cortisol shown in fig. 1 and 2 may be secondary to a strong effect on corticosterone. To better explore this phenomenon, the level of corticosterone in groups exhibiting the greatest changes in cortisol levels, e.g., groups treated with CDB-4124 at 20mg/kg or 10mg/kg, was determined. For comparison, the following groups were also analyzed: groups receiving 20mg/kg CDB-4124 plus 10mg/kg progesterone, groups receiving 10mg/kgRU 486, groups receiving only 10mg/kg progesterone, control groups. The level of corticosterone is 10-40 times higher than the level of cortisol. However, hardly any difference in the average corticosterone level was observed between the groups. No group differences were observed before treatment (p 0.43, Kruskal-Wallis test), after 21 days of treatment (p 0.57, Kruskal-Wallis test) or after 28 days of treatment and at kill (p 0.061, Kruskal-Wallis test).

To determine the effect of exogenous progesterone on serum corticosterone, corticosterone levels were compared in 3 paired groups that differed in whether they received exogenous progesterone (e.g., control versus progesterone, or CDB-4124, 20mg/kg versus CDB-4124, 20mg/kg plus progesterone, or CDB-4124, 10mg/kg versus CDB-4124, 10mg/kg plus progesterone). Statistically significant differences were detected: in animals treated with progesterone, corticosterone levels decreased after 21 days of treatment (p ═ 0.029, Mann-Whitney Wilcoxon test, two-tailed). This effect was not confirmed in sera taken at the time of killing. No difference in serum corticosterone was found between the progesterone and CDB-4124 groups, between the progesterone and RU-486 groups, or between RU-486 and CDB-4124 groups.

The relationship between cortisol and corticosterone in each group was also determined. For CDB-4124, 20mg/kg (r)²＝0.78)，CDB-4124，10mg/kg(r²0.82) and RU486 (r)²0.85), there is a strong positive linear relationship between cortisol and corticosterone. Addition of progesterone to the first two CDB-4124 groups greatly reduced the strength of the relationship (r for group 10²0.34 for group 11, r²0.3). Progesterone by itself does not show such a positive relationship (r)²1.0). The control group showed no relationship between these two glucocorticoids (r)²0.064). Thus, in the group receiving CDB-4124, increased cortisol levels were associated with corticosterone levels, perhaps due to some increase in conversion from corticosterone. This is consistent with the effect of CDB-4124 seen above: effects on metabolic enzymes responsible for progesterone and cortisol levels.

Although no strong effect of CDB-4124 on rat initial glucocorticoids was found, for safety reasons, patients administered CDB-4124 or CDB-4059 in phase I clinical trials should be monitored for possible anti-glucocorticoid effects, including possible increases in serum cortisol, corticosterone, or ACTH.

Example 5 determination of the antiproliferative Effect of SPRMs in uterine cells

Any uterine cell line may be used. Proliferation was measured in 96-well microtiter plates. Add 5X 10 to each well³And (4) cells. Media and drug solutions were added to the wells using Perkin Elmer Cetus PRO/PETTE. The medium was IMEM supplemented with 5% fetal bovine serum. 8 drug concentrations from 0.078uM to 10uM were determined in duplicate. The samples included tamoxifen alone, and each of the presently disclosed compounds in combination with tamoxifen.

After 4 days of culture, the medium was replaced with fresh medium containing the drug, and after a total of 7 days, the cell monolayer was fixed with trichloroacetic acid and stained with sulforhodamine dye. The absorbance (492nm) of the extracted dye solution was measured with a titetek Multiscan plate reader. Dose response curves (control absorbance vs. percent drug concentration) were constructed to estimate IC₅₀Value, IC₅₀Values are defined as the concentration of drug (micromolar) that inhibits proliferation by 50%. IC (integrated circuit)₅₀The values correlate with the efficacy of the test compound to inhibit cell proliferation, thus providing the information needed to identify compounds suitable for preventing uterine cell hyperproliferation.

Example 6 CDB-4124 reduces luteal phase progesterone in macaques

Macaques (Macaca fascicularis) (n ═ 14) were treated orally with CDB-4124 or RU-486 at 1.0 mg/kg/day or with placebo (control) for 36 weeks. Another group (n-14) received once a monthAnd IM. Uterine progesterone levels were measured in each animal during the middle of the trial (weeks 14-17) and the last month of the trial (weeks 33-36)1 month. The results are as follows:

luteal phase Progesterone reduction luteal phase Progesterone does not decrease

Control 113

13 1

RU 486 9 5

CDB-4124 8 6

Example 7 CDB-4124 does not reduce the Follicular Phase (Folliular Phase) estrogens in macaques

Uterine estrogen levels were determined for 1 month in each animal of example 6 during the middle of the trial (weeks 14-17) and the last month of the trial (weeks 33-36). The follicular phase results were based on 35 baseline ovulation cycles. The results are as follows:

mean Sd reduction?

Follicular phase 68.319.6

Control week 18 81.527.4 No

86323.8 th week no

Week 18 49.919.3 is

Week 36 41.713.4 is

RU486 no 67.427.1 at week 18

64.830.0 th week no

CDB-4124 week 18 63.824.6 No

67.322.9 th week no

Example 8 CDB-4124 andproliferation of endometrial epithelial cells of rhesus macaques was inhibited, but RU486 did not.

At week 36, 3 animals in each group of example 6 were injected with the thymidine analog bromodeoxyuridine (BrdU), a marker of proliferating cells and their progeny, within 24 hours of sacrifice to assess tissue proliferation. Uterus sections of intact thickness were stained and examined microscopically for evidence of proliferation, expressed as% of cells positive for BrdU incorporation:

uterus epithelial cell uterus stroma mammary gland

TXT Brdu-％ Brdu-％ Brdu-％

Control 10.0+ 2.52.6 + 0.62.4 +1.1

3.1+0.8 2.2+1.0 0.3+0.1

RU 486 12.6+1.8 3.1+1.0 0.9+0.3

CDB-4124 2.1+2.2 1.1+0.25 1.9+0.7

Example 9 CDB-4124 and RU486 increase apoptosis of endometrial epithelial cells of macaques, whileThe mortality was not increased.

Apoptosis in tissues from the same animals was assessed on glass slides by terminal deoxynucleotidyl transferase mediated dUTP-biotin nick end labeling (TUNEL) technique. The percentage of apoptotic cells was as follows:

uterus epithelial cell uterus stroma mammary gland

TXT apoptosis cell%

Control 0.2+ 0.10.7 + 0.20.5 +0.3

Lupron 0.2+0.1 0.2+0.1 1.4+0.7

RU 486 0.5+0.1 0.5+0.1 1.2+0.6

CDB-4124 0.5+0.2 0.5+0.1 2.6+0.9

Example 10 CDB-4124 inhibits human endometrial epithelial cell proliferation in a dose-dependent manner

39 premenopausal adult women diagnosed with endometriosis as Proelle in the treatment of endometriosisx^TM(CDB-4124) in a six month trial. The experiment included 3 dose levels of CDB-4124 and a positive control arm. The positive control isGnRH agonists, commonly used in the treatment of endometriosis (also known as endometrioses)). CDB-4124 was administered in a double-blind fashion as daily oral capsules at doses of 12.5 mg/day (n-2), 25 mg/day (n-3) and 50 mg/day (n-3) another group (n-4) was injected once a monthThe sustained release formulation of (3) was used as a positive control.

All doses of CDB-4124 andthe dose reduced the discomfort associated with pain on average over a 6 month period of drug exposure, the 50mg CDB-4124 dose more effectively reduced the duration and intensity of pain than the 12.5mg or 25mg dose, andin contrast, it was significantly better in terms of reduction of the number of painful days during the experiment. Comparison with ActivityPain relief also occurs more rapidly than pain relief. In this experiment, the response of pain to treatment was analyzed in two ways. In this experiment, patients maintained a daily pain diary to record pain severity and frequency. In addition, at each hospital visit, the patient filled out an endometriosis survey including a questionnaire that rated the pain intensity at bad days on a scale of 0-10, with 10 being the maximum intensity. The daily pain diary indicated that, in the first three months, the subjects receivedExperienced an average of 19.4 days of pain in women. Women receiving 50mg of CDB-4124 exhibited less than 1 day of pain during the first three months. And receiving the highest dose of CDB-4124 orWomen receiving 25mg and 12.5mg CDB-4124 showed more pain days than those recorded. This appears to be a dose-dependent effect on pain reduction. During the 180 day treatment period, the pain diary indicated that women receiving a 50mg dose of CDB-4124 had 170 or 96% pain free days (standard deviation of 8.86 days). This reduction in pain duration was statistically better than (p ═ 0.0012) acceptancePain free days reached 117.8 (74%; standard deviation 51.4 days). CDB-4124 at the 50mg dose was also statistically superior to the 25mg and 12.5mg doses on pain free days. Patients receiving doses of CDB-412412.5mg and 25mg had 115.9 (66%; standard deviation 69.2 days) and 133.6 (75%; standard deviation 27.4 days), respectively, pain free days. These results clearly support the dose response of CDB-4124. CDB-4124 at 25mg and 12.5mg doses withThere were no statistical differences. At the end of the first month of treatment, there was a statistically significant reduction in pain days compared to baseline in the 50mg proellex group (p ═ 0.031), but not in the other 3 treatment groups. Pain intensity was assessed by the following questions: "on a scale of 1-10, 0 is no pain, 10 is extreme pain, how strong your pain is on bad days? "mean score of baseline pain intensity is 6.3 for CDB-4124 group andgroup is 6.1. At 25mg and 50In the mg Proellex group, statistically significant pain relief was evident by the end of the first month. At month 3, all four active treatment groups had statistically significant pain reduction compared to baseline with the following scores: 3.7 for 12.5mg CDB-4124 (p ═ 0.03), 3.2 for 25mg CDB-4124 (p ═ 0.03), 1.6 for 50mg CDB-4124 (p ═ 0.015), andis 1.5(p ═ 0.016). These dose regimens were continued until month 6 when the pain intensity values were 2.0(p ═ 0.008), 2.8(p ═ 0.023), 0.6(p ═ 0.004) and 0.7(p ═ 0.016), respectively. Pain recurred 2 months after cessation of treatment and was of similar intensity in all four treatment groups.

In this experiment, by the end of month 3, the experiment was acceptedThe women in (a) showed on average a drop in estrogen to postmenopausal levels (< 20pg/ml) and remained until the 6 th month of treatment. This result is associated with a statistically significant increase in biomarkers of bone resorption compared to baseline at month 3 (p ═ 0.023), thus increasing the risk of bone loss. In the 6 th month and in the monthly follow-up visit (follow-up visit), the patient is in useThere was still an increase in bone resorption markers in treated women. CDB-4124 at all doses remained significantly higher than usedEstrogen concentrations observed and maintained within the low normal range (> 40pg/ml mean). Importantly, the biomarkers of bone resorption did not change any significantly in any dose arm of CDB-4124 at 3 and 6 months of treatment. Women with postmenopausal estrogen levels have been shown to be at greater risk of bone loss and other conditions. Due to the fact thatIn this way, the temperature of the molten steel is controlled,was not used for treatments lasting more than 6 months.

The side effects of CDB-4124 are generally mild, with no individual organ systems being systematically affected. Although this was a small experiment and no formal conclusions were drawn from the safety data, no single signal was observed.

In this experiment, changes in endometrial structure in women were closely monitored. Data from these examinations showed that CDB-4124 had an inverse dose-dependent effect on endometrial thickness during month 3. Two benchmarks were compared and one endometrial thickness ultrasound measurement was observed. After 3 months of treatment, women receiving a 50mg dose of CDB-4124 (n-3) showed no thickening of the endometrium and actually a tendency to decrease the thickness of the endometrium compared to baseline. 1 woman receiving a 25mg dose of CDB-4124 (n-4) and 2 women receiving a 12.5mg dose of CDB-4124 (n-4) showed endometrial thickening. ReceivingThe 5 women did not have endometrial thickening due to a low estrogen status. The results are as follows:

endometrium (mm)

Screening the first dose for 3 months

Lupron 7.9 7.5 2.75

CDB-4124(12.5mg) 7.5 8.0 20.33

CDB-4124(25mg) 8.4 11.7 19.6

CDB-4124(50mg) 8.0 10.8 7.7

In two cases of spotting and bleeding observed in patients with excessive endometrial thickening in the 12.5mg and 25mg CDB-4124 groups, dilation and curettage (D & C) procedures were performed to stop bleeding. No similar events were observed at the 50mg dose during the treatment period. After treatment cessation, greater than normal bleeding occurred in two patients in the 50mg CDB-4124 group, D & C procedures were performed in one patient, and the other patient was successfully conservatively controlled.

Claims (39)

1. A method of inhibiting endometrial hyperplasia, said method comprising administering to a female in need thereof an effective amount of a composition comprising a progesterone antagonist.

2. The method of claim 1, wherein the female in need thereof is a female with endometriosis.

3. The method of claim 1, wherein the composition is administered simultaneously, separately or sequentially with an estrogen or a selective estrogen receptor modulator as part of a hormone replacement therapy.

4. The method of claim 1, wherein the binding affinity of the progesterone antagonist to the progesterone receptor is at least 1.5 times greater than the binding affinity of the progesterone antagonist to the glucocorticoid receptor.

5. The method of claim 1, wherein progesterone levels in the female are not significantly increased after administration of the composition.

6. The method of claim 1, wherein estrogen levels in the female are not significantly reduced after administration of the composition.

7. The method of claim 1, wherein the progesterone antagonist is a compound of formula (I):

(I)

or a pharmaceutically acceptable salt, hydrate, or solvate thereof, wherein:

x represents alkyl, alkenyl, alkynyl, hydrogen, halogen, monoalkylamino or dialkylamino;

R₁represents O, NOH or NO-methyl;

R₂represents hydrogen or acetyl; and is

R₃Represents methoxy, formyloxy, acetoxy, acyloxy, S-alkoxy, acetylthienylmethyl, glycinate, vinyl ether, acetoxymethyl, methylcarbonate, halogen, methyl, hydroxy or ethoxy.

8. The method of claim 7, wherein said compound is CDB-4124.

9. The method of claim 8, wherein the compound is administered at a dose of 0.5mg/kg to 500 mg/kg.

10. The method of claim 9, wherein the compound is administered at a dose of 50 mg/day.

11. The method of claim 10, wherein the compound is administered for at least about 1 to about 6 months.

12. The method of claim 11, wherein the compound is administered for at least about 4 months.

13. A method of treating pain associated with endometriosis, said method comprising administering to a female in need thereof an effective amount of a composition comprising a progesterone antagonist.

14. The method of claim 13, wherein the progesterone antagonist is a compound of formula (I):

(I)

or a pharmaceutically acceptable salt, hydrate, or solvate thereof, wherein:

x represents alkyl, alkenyl, alkynyl, hydrogen, halogen, monoalkylamino or dialkylamino;

R₁represents O, NOH or NO-methyl;

R₂represents hydrogen or acetyl; and is

R₃Represents methoxy, formyloxy, acetoxy, acyloxy, S-alkoxy, acetylthienylmethyl, glycinate, vinyl ether, acetoxymethyl, methylcarbonate, halogen, methyl, hydroxy or ethoxy.

15. The method of claim 14, wherein said compound is CDB-4124.

16. The method of claim 13, wherein the size of endometrial lesions in the female are reduced.

17. The method of claim 13, wherein progesterone levels in the female are not significantly increased after administration of the composition.

18. The method of claim 13, wherein estrogen levels in the female are not significantly reduced after administration of the composition.

19. The method of claim 13, wherein the binding affinity of the progesterone antagonist for progesterone receptor is at least 1.5 times greater than the binding affinity of the progesterone antagonist for glucocorticoid receptor.

20. The method of claim 8, wherein the compound is administered at a dose of 0.5mg/kg to 500 mg/kg.

21. The method of claim 20, wherein the compound is administered at a dose of 50 mg/day.

22. The method of claim 21, wherein the compound is administered for at least about 1 to about 6 months.

23. The method of claim 22, wherein the compound is administered for at least about 4 months.

24. A method of treating an estrogen-dependent disorder in a female undergoing hormone therapy, the method comprising administering a progesterone antagonist to a female in need thereof.

25. The method of claim 24, wherein the disorder is endometrial hyperplasia or endometrial cancer.

26. The method of claim 24, wherein the progesterone antagonist is a compound of formula (I):

(I)

or a pharmaceutically acceptable salt, hydrate, or solvate thereof, wherein:

x represents alkyl, alkenyl, alkynyl, hydrogen, halogen, monoalkylamino or dialkylamino;

R₁represents O, NOH or NO-methyl;

R₂represents hydrogen or acetyl; and is

R₃Represents methoxy, formyloxy, acetoxy, acyloxy, S-alkoxy, acetylthienylmethyl, glycinate, vinyl ether, acetoxymethyl, methylcarbonate, halogen, methyl, hydroxy or ethoxy.

27. The method of claim 26, wherein said compound is CDB-4124.

28. The method of claim 26, wherein the hormone therapy is hormone replacement therapy comprising administration of estrogen.

29. The method of claim 26, wherein the hormone therapy comprises administration of a Selective Estrogen Receptor Modulator (SERM).

30. The method of claim 29, wherein the SERM is administered to treat a disease selected from the group consisting of: breast cancer, osteoporosis, colon cancer, neurodegenerative diseases such as parkinson's disease and alzheimer's disease, cardiovascular diseases, vaginal atrophy and obesity.

31. The method of claim 26, wherein the binding affinity of the compound for the progesterone receptor is at least 1.5 times greater than the binding affinity of the selective progesterone receptor modulator for the glucocorticoid receptor.

32. The method of claim 26, wherein progesterone levels in the female are not significantly increased after administration of the composition.

33. The method of claim 26, wherein the compound is administered at a dose of 0.5mg/kg to 500 mg/kg.

34. The method of claim 33, wherein the compound is administered at a dose of 50 mg/day.

35. The method of claim 34, wherein the compound is administered for at least 1 month.

36. The method of claim 34, wherein the compound is administered for at least 6 months.

37. The method of claim 34, wherein the compound is administered for at least 1 year.

38. The method of any one of claims 1, 13 or 24, wherein said composition is administered intermittently to said female, and wherein said female experiences menstruation during at least one discontinuation period.

39. The method of claim 38, wherein the administration period is about 4 months.