HK1065953B - The use of antigestagens in manufacturing a medicament for inhibiting the occurrence of advanced endometrium maturation in a female mammal - Google Patents

Description

Use of an antiprogestin for the production of a medicament for inhibiting accelerated endometrial maturation in a female mammal

This application is a continuation-in-part application of U.S. patent application No. 09/756,286, filed on 9/1/2001, the contents of which are incorporated herein by reference.

Technical Field

The present invention relates to the use of an antiprogestin for converting endometrial maturation after ovulation during (anti estagen) fertility treatment.

Background

In humans, gonadotropins are commonly used in the stimulation of the ovaries in assisted reproductive techniques, including in vitro fertilization and embryo transfer therapy (IVF/ET). However, post-ovulatory endometrial transition is promoted following controlled ovarian hyperstimulation (Garcia et al, 1984, Fertil. Steril.41: 31-37; Paulson et al, 1997, Fertil. Steril.76: 321-325; Kolb, 1997, Fertil. Steril.67: 625-630; Franchin et al, 1999, Fertil. Steril.71: 174-181). As a result, the temporary development of the endometrium and embryo, which are usually precisely synchronized, is disrupted, resulting in a low implantation rate of healthy blastocysts.

Disclosure of Invention

The present invention provides a method of enhancing fertility in a mammal by administering a 17 α -fluoralkylated progesterone receptor antagonist to inhibit accelerated endometrial maturation during fertility therapy (e.g., fertility enhancement), including all assisted reproductive therapy.

As part of the present invention, the method is particularly applicable to female mammals undergoing fertility treatment. The term "fertility treatment" as used herein refers to any treatment administered to a female mammal for the purpose of pregnancy, whether or not the mammal has been determined to be fertile, non-fertile, or otherwise impaired fertility. As part of the present invention, fertility treatments include, but are not limited to, Ovarian Hyperstimulation (OH), in vitro fertilization and embryo transfer (IVF/ET), and combinations of OH and IVF/ET. In particular, methods of inhibiting accelerated endometrial maturation are often performed in conjunction with ovarian hyperstimulation. For example, ovarian hyperstimulation may be used as a tool in fertility treatment in conjunction with, for example, timed sexual intercourse, artificial insemination, intrauterine insemination, IVF/ET, and partner intrafallopian tube transfer (GIFT).

IVF/ET in the present invention refers to any technique in which oocytes are transferred into the female reproductive tract after in vitro fertilization. For example, embryo transfer can be performed as follows: embryos are transferred to the uterus through the cervix or into the fallopian tubes (intrafallopian transfer of multiple Zygotes (ZIFT)). In vitro fertilization of an oocyte may be achieved, for example, by incubating the oocyte in the presence of sperm or by cytoplasmic sperm injection (ICSI). IVF/ET can also be achieved by using donor eggs. Fertility treatment also includes treatments such as the introduction of ovarian stimulation following normal coital insemination.

Ovarian hyperstimulation refers in the present invention to the use of a follicle stimulating agent to stimulate follicular development. Ovarian hyperstimulation may be used in female subjects with a variety of ovulation conditions, including subjects who do not ovulate, subjects with impaired, reduced or irregular ovulation, and subjects with normal ovulation behavior. Preferred follicle stimulating agents include gonadotropins such as Follicle Stimulating Hormone (FSH), Luteinizing Hormone (LH), human menopausal gonadotropin (hMG), and human chorionic gonadotropin (hCG). The known gonadotropins which can be used as follicle stimulating agents are for example the following trade names: perganol, Metrodin, Humegon, Fertinorm, Gonal F, and Primogonyl-100, and the like. In the present invention, the follicle stimulating agent also includes estrogen blockers such as clomiphene citrate (commercially available under the trade names Clomid and serolene). Gonadotropins which may be used according to the present invention include hormones isolated from natural sources as well as synthetic hormones, the latter including hormones prepared by recombinant DNA techniques. Also part of the invention are gonadotropins which retain activity as follicle stimulating agents.

Ovarian hyperstimulation can be accomplished by the use of various types of follicle stimulating agents. For example, estrogen blockers such as clomiphene may be used in combination with gonadotropins. Ovarian hyperstimulation can be used to treat various types of infertility, including but not limited to, idiopathic infertility, anovulatory infertility, infertility associated with endometriosis, fallopian tube factor infertility, and male factor infertility.

Ovarian hyperstimulation can also be accomplished in conjunction with the use of gonadotropin releasing hormone antagonists (GnRH) to shut down endogenous hormone formation in a subject. GnRH which may be used in conjunction with the methods of the present invention include, for example, Synarel or Lupron.

In a preferred embodiment, the method of the invention is for use in a female subject. The methods of the invention may also be used in other female mammals, including but not limited to pets, farm animals, and zoo animals, including particularly cattle, swine, horses, and sheep. If used in fertility treatment of non-human mammals, administration of an antiprogestin may be helpful to achieve a higher success rate in vitro fertilization and embryo transfer, for example for economic or reproductive purposes.

Non-human mammals produced by the methods of the invention are also part of the invention. Such non-human mammals include, for example, mammals that have been genetically modified, including, for example, mammals that have been genetically modified using recombinant DNA techniques.

In one aspect, the present invention relates to a method of inhibiting the occurrence of accelerated endometrial maturation in a female subject undergoing fertility treatment, comprising administering to said female subject at least one 17 α -fluoralkylated progesterone receptor antagonist during the post-ovulatory phase of the endometrial cycle.

In another aspect, the invention relates to a method of achieving pregnancy in a female subject, comprising stimulating the ovary of the subject by administering to the subject a follicle stimulating agent, wherein the stimulating agent comprises follicle stimulating hormone; removing ova from the stimulated ovary of the subject; administering to the subject at least one 17 α -fluoralkylated progesterone receptor antagonist in the post-ovulatory phase of the endometrial cycle; fertilizing at least one ovum in vitro to obtain an embryo; the embryo is implanted into the uterus or fallopian tube of said subject.

In another aspect, the invention relates to a method of inhibiting the occurrence of accelerated endometrial maturation in a non-human female mammal undergoing fertility treatment to achieve pregnancy, comprising administering to said mammal at least one 17 α -fluoralkylated progesterone receptor antagonist in the post-ovulatory phase of the endometrial cycle.

In another aspect, the invention relates to a method of achieving pregnancy in a non-human mammal, comprising stimulating the ovary of the mammal by administering to the mammal a follicle stimulating agent, wherein the stimulating agent comprises follicle stimulating hormone; removing the ova from the ovaries of the stimulated mammal; administering to the mammal at least one 17 α -fluoralkylated progesterone receptor antagonist in the post-ovulatory phase of the endometrial cycle; fertilizing at least one ovum in vitro to obtain an embryo; the embryo is implanted into the uterus or fallopian tube of said mammal.

In another aspect, the present invention relates to a method of achieving pregnancy in a female mammal, comprising:

(a) administering to said mammal a gonadotropin which is over stimulated for the ovary;

(b) removing the ova from the stimulated mammal;

(c) administering at least one 17 α -fluoralkylated progesterone receptor antagonist;

(d) fertilizing at least one ovum in vitro to obtain an embryo;

(e) introducing the embryo into the reproductive tract of said mammal.

Preferably, the gonadotropins administered to the mammal to achieve ovarian hyperstimulation comprise Follicle Stimulating Hormone (FSH). More preferably, ovarian stimulation is achieved by the use of a first gonadotropin comprising FSH followed by the use of a second gonadotropin comprising chorionic gonadotropin, in particular human chorionic gonadotropin (hCG). The embryo is preferably introduced into the uterus via the cervix and thus into the reproductive tract of the mammal. It is also preferred that the gonadotropin releasing hormone (GnRH) agonist or antagonist is administered to the female mammal prior to and during administration of the gonadotropin.

The compounds which can be used according to the invention as antiprogestins include all 17 α -fluoralkylated progesterone receptor antagonists which have a strong affinity for the gestagen receptor (progesterone receptor) and which themselves have minimal gestagen activity. For example, 17 α -fluoroalkylated steroids useful in the present invention are described in the following documents: U.S. patent application No. 09/020,947, WO 98/34947, Wang and Ruan, 1994, Journal of Fluorine Chemistry 69: 1-3. The contents of these documents are incorporated herein by reference. Antiprogestins useful in the present invention include, but are not limited to, 17 α -fluoroalkylated steroids of the general formula I:

wherein:

R¹is a methyl group or an ethyl group,

R²is C_nF_mH_oWherein n is 1 to 6, preferably 2, 3, 4,5 or 6, m > 1 and m + o is 2n + 1;

R³is a free, etherified or esterified hydroxy group;

R⁴and R⁵Are each hydrogen, or together form an additional bond or methylene;

st is a steroid ABC ring system represented by the following partial formula A, B or C:

in the formula:

R⁶is hydrogen, straight chain C_1-4Alkyl or branched C_3-4Alkyl or halogen;

R⁷is hydrogen, straight chain C_1-4Alkyl or branched C_3-4An alkyl group; or

If St is part of a steroidal ABC ring system of the formula A or B, then in addition

R⁶And R⁷May together form additional bonds;

x is oxygen, hydroxyimino (═ N-OH), or two hydrogen atoms;

R⁸is Y or optionally at a plurality of positionsAryl substituted with a group Y other than hydrogen;

y is hydrogen, halogen, -OH, -NO₂、-N₃、-CN、-NR^9aR^9b、-NHSO₂R⁹、-CO₂R⁹、C_1-10Alkyl radical, C_1-10Alkoxy radical, C_1-10Alkanoyloxy, benzoyloxy, C_1-10Alkanoyl radical, C_1-10Hydroxyalkyl or benzoyl;

R^9aand R^9bMay be the same or different and represents hydrogen or C_1-10An alkyl group;

R⁹is hydrogen or C_1-10An alkyl group;

for-NR^9aR^9bGroups and also their pharmaceutically acceptable salts with acids, and for those in which R is⁹of-CO as hydrogen₂R⁹Also included are pharmaceutically acceptable salts thereof with bases.

The broken lines indicate that the indicated groups may be in the alpha-or beta-position.

In the above formula, suitable alkyl groups include methyl, ethyl, n-propyl or isopropyl, and n-butyl, isobutyl, or tert-butyl.

Others C_1-10Alkyl radical Y, R⁹、R^9a、R^9bHigher homologues such as pentyl, neopentyl, and hexyl up to decyl are included.

However, C_1-10Alkyl is to be understood as also including carbocyclic or alkylcycloalkyl groups having up to 10 carbon atoms, such as cyclopropyl, cyclopentyl, cycloheptyl, methylcyclopropyl, methylcyclopentyl or methylcyclohexyl. Methyl and ethyl are preferred.

C_1-10Alkoxy is understood to mean a radical which is derived from the abovementioned alkyl radicals and is extended by an oxygen atom, such as methoxy, ethoxy, n-propoxy or isopropoxy, and also n-butoxy, isobutoxy, orT-butoxy group.

C_1-10Alkanoyl is defined as being derived from straight or branched chain C_1-10Acyl groups of alkyl carboxylic acids such as formyl, acetyl, propionyl, butyryl, isobutyryl, and the like.

C_1-10The alkanoyloxy group means a group in which the above-mentioned alkanoyl group is extended with one oxygen atom, and examples thereof include acetoxy, propionyloxy, and butyryloxy groups.

If halogen atoms are used as substituents, they may be fluorine, chlorine or bromine atoms. Preferably fluorine.

For the group R²Perfluorinated side chains of length n-2-4 are preferred, with pentafluoroethyl units being particularly preferred.

R³Mainly representing free hydroxyl groups.

If etherified or esterified hydroxy is used as the 17 beta-substituent, the latter is preferably C_1-10By etherification of alkyl radicals or with C_1-10And (4) esterifying alkanoyl. As for the alkyl group or the alkanoyl group, their meanings are the same as those described above. Etherification or esterification of the hydroxyl groups can be carried out according to analogous methods known to the person skilled in the art.

R⁴And R⁵Preferably hydrogen respectively or together form an additional bond.

If R is⁸Is a group Y, which is preferably C_1-10Alkanoyl, or (1-hydroxy) -C_1-10An alkyl group; among them, acetyl and propionyl are particularly preferable.

Preferred carbocyclic or heterocyclic aryl radicals are phenyl, 1-or 2-naphthyl, 2-or 3-furyl, 2-or 3-benzofuryl, 2-or 3-thienyl, 2-, 3-or 4-pyridyl. Substituted aryl radicals R⁸Predominantly 4-cyanophenyl and 4-halophenyl radicals, in particular the 4-fluorophenyl radical.

In all of the above R⁸In the preferred group of (1), R⁸Has the meaning of Y, with Y again being particularly preferably acetyl.

The compounds described below are particularly preferred for the present invention:

11 β - (4-acetylphenyl) -17 β -hydroxy-17 α - (1, 1, 2, 2, 2-pentafluoroethyl) -estr-4-en-3-one;

4 '- [17 β -hydroxy-3-oxo-17 α - (1, 1, 2, 2, 2-pentafluoroethyl) estr-4-en-11 β -yl ] [1, 1' -biphenyl ] -4-carbonitrile;

11 β - (4 '-fluoro [1, 1' -biphenyl ] -4-yl) -17 β -hydroxy-17 α - (1, 1, 2, 2, 2-pentafluoroethyl) -estr-4-en-3-one;

17 β -hydroxy-17 α - (1, 1, 2, 2, 2-pentafluoroethyl) -11 β - [4- (3-pyridyl) phenyl ] estr-4-en-3-one;

11 β - (4-acetylphenyl) -17 β -hydroxy-17 α - (1, 1, 2, 2, 2-pentafluoroethyl) -estra-4, 15-dien-3-one;

4 '- [17 β -hydroxy-3-oxo-17 α - (1, 1, 2, 2, 2-pentafluoroethyl) estra-4, 15-dien-11 β -yl ] [1, 1' -biphenyl ] -4-carbonitrile;

11 β - (4 '-fluoro [1, 1' -biphenyl ] -4-yl) -17 β -hydroxy-17 α - (1, 1, 2, 2, 2-pentafluoroethyl) -estra-4, 15-dien-3-one;

17 β -hydroxy-17 α - (1, 1, 2, 2, 2-pentafluoroethyl) -11 β - [4- (3-pyridyl) phenyl ] estra-4, 15-dien-3-one;

11 β - (4-acetylphenyl) -17 β -hydroxy-17 α - (1, 1, 2, 2, 2-pentafluoroethyl) -estra-4, 9-dien-3-one;

4 '- [17 β -hydroxy-3-oxo-17 α - (1, 1, 2, 2, 2-pentafluoroethyl) estra-4, 9-dien-11 β -yl ] [1, 1' -biphenyl ] -4-carbonitrile;

11 β - (4 '-fluoro [1, 1' -biphenyl ] -4-yl) -17 β -hydroxy-17 α - (1, 1, 2, 2, 2-pentafluoroethyl) -estra-4, 9-dien-3-one;

17 β -hydroxy-17 α - (1, 1, 2, 2, 2-pentafluoroethyl) -11 β - [4- (3-pyridyl) phenyl ] estra-4, 9-dien-3-one;

11 β - (4-acetylphenyl) -17 β -hydroxy-17 α - (1, 1, 2, 2, 2-pentafluoroethyl) -estra-4, 9, 15-trien-3-one;

4 '- [17 β -hydroxy-3-oxo-17 α - (1, 1, 2, 2, 2-pentafluoroethyl) estra-4, 9, 15-trien-11 β -yl ] [1, 1' -biphenyl ] -4-carbonitrile;

11 β - (4 '-fluoro [1, 1' -biphenyl ] -4-yl) -17 β -hydroxy-17 α - (1, 1, 2, 2, 2-pentafluoroethyl) -estra-4, 9, 15-trien-3-one;

17 β -hydroxy-17 α - (1, 1, 2, 2, 2-pentafluoroethyl) -11 β - [4- (3-pyridyl) phenyl ] estra-4, 9, 15-trien-3-one;

6 ' -acetyl-9, 11 α -dihydro-17 β -hydroxy-17 α - (1, 1, 2, 2, 2-pentafluoroethyl) -4 ' H-naphthalene [3 ', 2 ', 1 ': 10, 9, 11] estr-4-en-3-one;

4- [9, 11 α -dihydro-17 β -hydroxy-3-oxo-17 α - (1, 1, 2, 2, 2-pentafluoroethyl) -4 'H-naphthalene [ 3', 2 ', 1': 10, 9, 11] estr-4-en-6' -yl ] benzonitrile;

9, 11 α -dihydro-6 ' - (4-fluorophenyl) -17 β -hydroxy-17 α - (1, 1, 2, 2, 2-pentafluoroethyl) -4 ' H-naphthalene [3 ', 2 ', 1 ': 10, 9, 11] estr-4-en-3-one;

9, 11 α -dihydro-17 β -hydroxy-17 α - (1, 1, 2, 2, 2-pentafluoroethyl) -6 ' - (3-pyridyl) -4 ' H-naphthalene [3 ', 2 ', 1 ': 10, 9, 11] estr-4-en-3-one;

6 ' -acetyl-9, 11 α -dihydro-17 β -hydroxy-17 α - (1, 1, 2, 2, 2-pentafluoroethyl) -4 ' H-naphthalene [3 ', 2 ', 1 ': 10, 9, 11] estra-4, 15-dien-3-one;

4- [9, 11 α -dihydro-17 β -hydroxy-3-oxo-17 α - (1, 1, 2, 2, 2-pentafluoroethyl) -4 'H-naphthalene [ 3', 2 ', 1': 10, 9, 11] estra-4, 15-dien-6' -yl ] benzonitrile;

9, 11 α -dihydro-6 ' - (4-fluorophenyl) -17 β -hydroxy-17 α - (1, 1, 2, 2, 2-pentafluoroethyl) -4 ' H-naphthalene [3 ', 2 ', 1 ': 10, 9, 11] estra-4, 15-dien-3-one;

9, 11 α -dihydro-17 β -hydroxy-17 α - (1, 1, 2, 2, 2-pentafluoroethyl) -6 ' - (3-pyridyl) -4 ' H-naphthalene [3 ', 2 ', 1 ': 10, 9, 11] estra-4, 15-dien-3-one;

17 β -hydroxy-11 β - (4-hydroxyphenyl) -17 α - (1, 1, 2, 2, 2-pentafluoroethyl) estra-4, 9-dien-3-one;

17 β -hydroxy-11 β - (4-hydroxyphenyl) -17 α - (1, 1, 2, 2, 2-pentafluoroethyl) estr-4-en-3-one;

9, 11 α -dihydro-6 ', 17 β -dihydroxy-17 α - (1, 1, 2, 2, 2-pentafluoroethyl) -4 ' H-naphthalene [3 ', 2 ', 1 ': 10, 9, 11] estr-4-en-3-one;

11 β - [4- (acetoxy) phenyl ] -17 β -hydroxy-17 α - (1, 1, 2, 2, 2-pentafluoroethyl) estra-4, 9-dien-3-one;

11 β - [4- (acetoxy) phenyl ] -17 β -hydroxy-17 α - (1, 1, 2, 2, 2-pentafluoroethyl) estr-4-en-3-one;

6 ' -acetoxy-9, 11 α -dihydro-17 β -hydroxy-17 α - (1, 1, 2, 2, 2-pentafluoroethyl) -4 ' H-naphthalene [3 ', 2 ', 1 ': 10, 9, 11] estr-4-en-3-one;

17 β -hydroxy-11 β - [4- (hydroxymethyl) phenyl ] -17 α - (1, 1, 2, 2, 2-pentafluoroethyl) estra-4, 9-dien-3-one;

17 β -hydroxy-11 β - [4- (hydroxymethyl) phenyl ] -17 α - (1, 1, 2, 2, 2-pentafluoroethyl) estr-4-en-3-one;

9, 11 α -dihydro-17 β -hydroxy-6 ' -hydroxymethyl-17 α - (1, 1, 2, 2, 2-pentafluoroethyl) -4 ' H-naphthalene [3 ', 2 ', 1 ': 10, 9, 11] estr-4-en-3-one;

4- [17 β -hydroxy-3-oxo-17 α - (1, 1, 2, 2, 2-pentafluoroethyl) estra-4, 9-dien-11 β -yl ] benzaldehyde;

4- [17 β -hydroxy-3-oxo-17 α - (1, 1, 2, 2, 2-pentafluoroethyl) estr-4-en-11 β -yl ] benzaldehyde;

9, 11 α -dihydro-17 β -hydroxy-3-oxo-17 α - (1, 1, 2, 2, 2-pentafluoroethyl) -4 'H-naphthalene [ 3', 2 ', 1': 10, 9, 11] estr-4-en-6' -al;

methyl 4- [17 β -hydroxy-3-oxo-17 α - (1, 1, 2, 2, 2-pentafluoroethyl) estra-4, 9-dien-11 β -yl ] benzoate;

4- [17 β -hydroxy-3-oxo-17 α - (1, 1, 2, 2, 2-pentafluoroethyl) estr-4-en-11 β -yl ] benzoic acid methyl ester;

9, 11 α -dihydro-17 β -hydroxy-3-oxo-17 α - (1, 1, 2, 2, 2-pentafluoroethyl) -4 'H-naphthalene [ 3', 2 ', 1': 10, 9, 11] estr-4-ene-6' -carboxylic acid methyl ester;

17 β -hydroxy-11 β - [4- (1-hydroxyethyl) phenyl ] -17 α - (1, 1, 2, 2, 2-pentafluoroethyl) estra-4, 9-dien-3-one;

17 β -hydroxy-11 β - [4- (1-hydroxyethyl) phenyl ] -17 α - (1, 1, 2, 2, 2-pentafluoroethyl) estr-4-en-3-one;

9, 11 α -dihydro-17 β -hydroxy-6 ' - (1-hydroxyethyl) -17 α - (1, 1, 2, 2, 2-pentafluoroethyl) -4 ' H-naphthalene [3 ', 2 ', 1 ': 10, 9, 11] estr-4-en-3-one.

Preferred is 11 β - (4-acetylphenyl) -17 β -hydroxy-17 α - (1, 1, 2, 2, 2-pentafluoroethyl) -estra-4, 9-dien-3-one (═ compound a).

The antiprogestin can be administered, for example, by oral, topical, enteral or parenteral routes.

For preferred oral administration, particularly suitable dosage forms are tablets, dragees, capsules, pills, suspensions or solutions, which can be prepared by conventional methods with additives and carriers used in the pharmaceutical field. For topical administration, for example, a vaginal ring or a transdermal system such as a dermal plaster may be used. For parenteral administration, particularly suitable are solutions, preferably oily or aqueous solutions, and suspensions or emulsions. Ampoules are convenient unit doses.

According to a preferred embodiment of the invention, the 17 α -fluoralkylated progesterone receptor antagonist is generally administered during the period of 1 to 6 days, preferably 1 to 4 days, more preferably 1 to 3 days after ovulation and/or removal of the oocyte. Typically, the antiprogestin is administered, for example, 1-6 days, preferably 1-4 days, more preferably 1-3 days after ovulation induction with chorionic gonadotropin.

Surprisingly, it has been found according to the present invention that low doses of 17 α -fluoralkylated progesterone receptor antagonists are effective in the methods of the present invention. The 17 α -fluoralkylated progesterone receptor antagonist is preferably administered in a daily dosage of up to 10mg, preferably 0.1-2mg, more preferably 0.1-1mg, and most preferably 0.1-0.7mg per female subject. For mammals, the daily dose is usually 0.01-1mg/kg, preferably 0.01-0.3mg/kg, and more preferably 0.01-0.1 mg/kg. The daily dose of the antiprogestin can be administered in a single dose or in multiple divided doses per day.

In a preferred embodiment according to the invention, the amount of antiprogestin administered to a human subject in a single day is 0.1-2mg, more preferably 0.1-1mg, and most preferably 0.1-0.7mg per subject; and the amount to the mammal is 0.01 to 1mg/kg, preferably 0.01 to 0.3mg/kg, more preferably 0.01 to 0.1 mg/kg. For example, administration is performed on days 1, 2, 3, 4,5 or 6, preferably days 1-3, more preferably day 2 after ovulation, oocyte retrieval, or administration of chorionic gonadotropin.

For any particular 17 α -fluoralkylated progesterone receptor antagonist, the most suitable dosage can be determined as described by Herrmann, w. et al (1982, complexes Rendus 294: 933), for example, by assessing the efficacy of inducing premature menstruation in the luteal-late phase of the human cycle. Use of antiprogestins in delaying endometrial maturation is described in U.S. patent No. 4,764,513, EP 0219447B1 and Hegele-Hartung et al, 1992, Endocrinology 131: 2446 and 2460, the contents of which are incorporated herein by reference.

The compounds of the invention may be employed in admixture with conventional excipients, for example, pharmaceutically acceptable organic or inorganic carrier materials suitable, for example, for parenteral, enteral (e.g., oral) or topical administration without deleteriously reacting with the active compound. Suitable pharmaceutically acceptable carriers include, but are not limited to, water, salt solutions, alcohols, gum arabic, vegetable oils, benzyl alcohol, polyethylene glycol, gelatin, sugars such as lactose, amylase or starch, magnesium stearate, talc, silicic acid, viscous paraffin, essential oils, fatty acid mono-and diglycerides, pentaerythritol fatty acid esters, hydroxymethyl cellulose, polyvinylpyrrolidone, and the like. The pharmaceutical preparations can be sterilized and, if desired, mixed with auxiliary agents which react nondestructively with the active compounds, such as lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure, buffers, colorants, flavors and/or fragrances and the like. They may also be mixed with other active agents, such as vitamins, if desired.

Sustained or direct release compositions can be formulated, such as liposomes or those in which the active ingredient is protected with a coating having a different degradability, such as microencapsulation or multilayer coating, etc. It is also possible to lyophilize the novel compounds and to use the lyophilizates obtained, for example, for the preparation of injectable products.

For topical administration, non-spray, viscous to semi-solid or solid dosage forms may be used, which may include carriers compatible with topical administration and which have a dynamic viscosity preferably greater than water. Suitable formulations include, but are not limited to, solutions, suspensions, emulsions, creams, ointments, powders, liniments, salves, sprays, and the like, which can be sterilized or mixed with adjuvants, such as preservatives, stabilizers, wetting agents, buffers, or salts that affect osmotic pressure, and the like, if desired. For topical administration, sprays which can be administered by spraying are also suitable, in which the active ingredient, in particular in admixture with a solid or liquid inert carrier material, is packaged in a squeeze bottle or with a propellant which is volatile under pressure, normally a gas, such as freon.

It will be appreciated that the actual preferred amount of active compound in a particular instance will vary depending upon the particular compound used, the particular composition formulated, the form of administration, and the particular site and species to be treated. For a given host, dosages may be determined using conventional methods, e.g., by routine comparison of test compounds to known drugs for different activities using appropriate and conventional pharmacological protocols.

Drawings

FIG. 1 is a schematic explanatory view of the experiment described in example 1. The day of administration of human chorionic gonadotropin (hCG) was defined as day 0 of pseudopregnancy (d 0). As shown in the figure, ovarian stimulation in animals occurred at-d 3, -d2, and-d 1 induced by pseudopregnancy, while progesterone receptor antagonists were administered on day 2 of pseudopregnancy (d 2).

Detailed Description

Without further elaboration, it is believed that one skilled in the art can, using the preceding description, utilize the present invention to its fullest extent. The following preferred specific embodiments are, therefore, to be construed as merely illustrative, and not a limitation of the scope of the present invention.

In the above description and in the following examples, all temperatures are in degrees celsius; and all parts and percentages are by weight unless otherwise indicated.

Example 1: increasing endometrial receptivity in a rabbit model of overovulation with low doses of 17 alpha-fluoralkylated progesterone receptor antagonists

The effect of 17 alpha-fluoralkylated progesterone receptor antagonists on endometrial activity was evaluated in a rabbit model of ovarian hyperstimulation. Pseudopregnancy in rabbits has been demonstrated to be a model system for the luteal phase in humans (Beier and Kuhnel, 1973, Hormone Res.4: 1-27; Fischer et al, 1985, Fertilitat 1: 101-. In this species, ovarian stimulation may induce acceleration of secretory conversion (Delbos-Winter et al, 1987, Fertilitat 3: 87-93) and enhancement of glandular formation.

A total of 25 sexually mature but unproductive New Zealand white rabbits (body weight 3.1-4.0kg) were used for the analysis. Animals were divided into 2 control groups (groups 1 and 2) and 3 treatment groups (groups 3-5), with 5 animals in each group.

Control group 1 received 75 International Units (IU) of human chorionic gonadotropin (hCG), which was administered intravenously to induce pseudopregnancy. The day of hCG injection was defined as day 0 of pseudopregnancy (d0 p.

Control group 2 and treatment groups 3, 4 and 5 were treated with 5IU of Human Menopausal Gonadotropin (HMG) personal^Gonadotropin stimulation was performed for 3 days (Serono Pharma, Unterschleissheim, Germany), which were administered by subcutaneous injection to induce multiple follicle growth. Pseudopregnancy was induced by administration of human chorionic gonadotropin (hCG) as described above for control group 1.

Treatment groups 3, 4 and 5 also received a single oral administration of a 17 α -fluoralkylated progesterone receptor antagonist on day 2 (d2 p.hcg) after induction of pseudopregnancy by administration of human chorionic gonadotropin. The specific treatment protocol is shown in figure 1.

Animals in all treatment groups were sacrificed on day 5 of pseudopregnancy induction (d5 p.hcg) and the uterus removed. The relative wet weight of uterus (mg/100g body weight) was determined and a portion of the uterus was frozen in liquid nitrogen for determining the expression of uteroglobin by in situ hybridization. The expression of uteroglobin is known as a highly sensitive parameter for determining endometrial receptivity (Beier, H.M., 1968, Biochem Biophys.acta.160: 289) 291; Beier, H.M., 1982, in: Beier, H.M., and Karlson, P. (eds.), Proteins and Steroids in early Pregnancy), and as a marker for secretory activity (Beier, 1976, J.reprod.Fertil.Suppl.25: 53-69) and for differences in endometrial epithelium (Krusch & Beier, 1994, Ann.Ant.176: 23-31). In situ hybridization for detecting uteroglobin expression was performed as described in Krusche & Beier, supra. Another uterus was removed and processed for paraffin histology to determine endometrial transition by using the McPhail index (McPhail MK, 1934, J.Phys.83: 145-156). The McPhail index reflects varying degrees of gestagen (conversion) activity in rabbit endometrium:

1. no transformation

2. Low transition

3. Obvious transformation

4. High transition

The results obtained are shown in table 1 below.

Table 1: effect of 17 alpha-Fluoroalkylated progesterone receptor antagonist monotherapy on uterine weight, endometrial transitions and uteroglobin mRNA expression at day 5 (d5 p.hCG) after pseudopregnancy induction in superovulated pseudopregnant rabbits

Group of	Treatment regimens			Measurement endpoint at d5 p.hCG
							-d3，-d2，-d1p.hCG	d0p.hCG	d2p.hCG
	5IUHMG	75IUHCG	Progesterone receptor antagonist [ mg/kg]							Wet weight of uterus [ mg/kg][x±SD]	Mc-Phail[x±SD]	Uteroglobin ISH [ expression in luminal epithelial cells]
				1	-	+	-	324±99	3.2±0.2				Very high expression
2	+	+	0							448±80	3.9±0.1	Very low expression
				3	+	+	0.1	305±48	3.0±0.1				Very high watchTo achieve
4	+	+	1							227±56	1.5±0.3	Very low expression
				5	+	+	10	144±30	1.1±0.3				Is not provided with

As shown in the table, animals that were ovarian stimulated but not treated with anti-gestagen (group 2) showed accelerated endometrial maturation, as evidenced by nearly undetectable uteroglobin expression in endometrial cavity epithelial cells, uterine weight, and a relatively high Mc-phase index. However, animals that were over-ovarian stimulated and treated with antiprogestin (groups 3-5) demonstrated delayed endometrial maturation as evidenced by uterine wet weight and the Mc-Phail index. Endometrial maturation for animals treated with low dose progesterone receptor antagonist (0.1mg/kg) (group 3) was approximately comparable to control animals that were not over-stimulated by ovary (group 1). In contrast, higher doses of progesterone receptor antagonist, 1mg/kg (group 4) and 10mg/kg (group 5), significantly offset endometrial maturation associated with ovarian hyperstimulation, and also delayed the time to maturation compared to animals without ovarian hyperstimulation (group 1).

These examples can be repeated with similar success, substituting conventional or specific reactants and/or operating conditions for those used in the examples above.

From the foregoing description, one skilled in the art can easily ascertain the essential characteristics of this invention, and without departing from the spirit and scope thereof, can make various changes and modifications of the invention to adapt it to various usages and conditions.

Claims (6)

1. Use of a compound of formula I for the manufacture of a medicament for inhibiting accelerated endometrial maturation in a female mammal:

wherein:

R¹is a methyl group or an ethyl group,

R²is C_nF_mH_oWherein n is 1-6, m > 1, and m + o is 2n + 1;

R³is a free, etherified or esterified hydroxy group;

R⁴and R⁵Are each hydrogen, or together form an additional bond or methylene;

st is a steroid ABC ring system represented by the following partial formula A, B or C:

in the formula:

R⁶is hydrogen, straight chain C_1-4Alkyl or branched C_3-4Alkyl or halogen;

R⁷is hydrogen, straight chain C_1-4Alkyl or branched C_3-4An alkyl group; or

If St is part of a steroidal ABC ring system of the formula A or B, then in addition

R⁶And R⁷May together form additional bonds;

x is oxygen, hydroxyimino, or two hydrogen atoms;

R⁸is Y or aryl optionally substituted at various positions with a group Y other than hydrogen;

y is hydrogen, halogen, -OH, -NO₂、-N₃、-CN、-NR^9aR^9b、-NHSO₂R⁹、-CO₂R⁹、C_1-10Alkyl radical, C_1-10Alkoxy radical, C_1-10Alkanoyloxy, benzoyloxy, C_1-10Alkanoyl radical, C_1-10Hydroxyalkyl or benzoyl;

R^9aand R^9bMay be the same or different and represents hydrogen or C_1-10An alkyl group;

R⁹is hydrogen or C_1-10An alkyl group;

for-NR^9aR^9bGroups and also their pharmaceutically acceptable salts with acids, and for those in which R is⁹of-CO as hydrogen₂R⁹Also included are pharmaceutically acceptable salts thereof with bases.

2. Use according to claim 1, wherein n is 2, 3, 4,5 or 6.

3. The use of claim 1, wherein the medicament is in an orally administered dosage form.

4. The use of claim 1, wherein the medicament further comprises follicle stimulating hormone.

5. The use of claim 1, wherein the mammal is undergoing fertility enhancing treatment to achieve pregnancy.

6. The use according to any one of claims 1 to 5, wherein the mammal comprises a human.