JP7141048B2 - Estrogen preparations for estrogen replacement therapy using halogenated estrogens - Google Patents

Description

The present invention relates to estrogen preparations for estrogen replacement therapy containing halogenated estrogens as active ingredients. More particularly, the present invention relates to estrogen formulations for estrogen replacement therapy containing non-carcinogenic halogenated estrogens as active ingredients.

A woman's life cycle is dramatically affected as she ages. In particular, estrogen produced by the ovaries due to menopause starting around the age of 45 not only regulates the functions of the reproductive organs, but also exerts a wide range of physiological effects on the cardiovascular, skeletal, and nervous systems. Appropriate medical care is required because the decline in estrogen production associated with menopause causes various menopausal symptoms, indefinite complaints, and osteoporosis.

In Europe and the United States, hormone replacement therapy (HRT) containing estrogen as a main ingredient is widely used to improve women's menopause or osteoporosis (Non-Patent Document 1). However, many epidemiological studies show that long-term estrogen administration increases the incidence of breast cancer (Non-Patent Documents 2 to 4), ovarian cancer (Non-Patent Document 5), and endometrial cancer (Non-Patent Document 6). has become clear. For this reason, estrogen therapy carries the risk of secondary carcinogenesis. Even in Japan, there are millions of people to be treated, but there is a strong tendency to shy away from treatment because of its side effects. Therefore, in order to improve the quality of life of middle-aged and elderly women and promote active participation in society, it is essential to develop a safe alternative estrogen without carcinogenic risk.

Human estrogen (estradiol: E ₂ and estrone: E ₁ ) is the main component of estrogen preparations used as hormone replacement therapy (Non-Patent Document 7). In the metabolic process after being absorbed into the body, these compounds are hydroxylated at the 2- or 4-position and become catechol estrogens (Fig. 1). This catechol estrogen is further oxidized and metabolized into catechol estrogen quinone, and the oxidative radicals generated in this process cause oxidative damage to DNA in the body (Non-Patent Document 8). In addition, the produced quinone also binds directly to DNA and causes adduct damage (Non-Patent Document 9). The inventors discovered that these DNA oxidative damage and adduct damage induce gene mutations during the process of DNA replication (Non-Patent Documents 10 to 12), and have already published papers and the like. In fact, DNA oxidative damage has been detected in mammary glands of breast cancer patients (Non-Patent Document 13). Therefore, it has been clarified that these gene lesions are deeply involved in the carcinogenesis process of breast cancer.

August Copenhagen Irish (ACI) rats have pathologically similar mammary gland tissue to humans, and therefore are regarded as very useful model animals for research on human breast cancer (Non-Patent Document 14). In fact, it has been reported that subcutaneous administration of a small amount of estrogen-containing pellets to ACI rats causes breast cancer several months later (Non-Patent Documents 14 and 15). We have confirmed the frequency of breast cancer carcinogenesis (Non-Patent Document 16). From these facts, it is clear that receiving estrogen-containing hormone replacement therapy involves the risk of carcinogenesis such as breast cancer.

Long-term administration of estrogen has been reported to cause kidney cancer in male Syrian hamsters (Non-Patent Document 17). Fluorinated estrogen was used to investigate the mechanism, and 4-fluoro-estradiol (4-FE ₂ ) was found to be carcinogenic, while 2-FE ₂ was not found to be carcinogenic (Non-Patent Documents 18 and 19 ). However, the cancerous organ caused by the administration of fluorinated estrogen was the kidney, and it was not a cancer peculiar to women, including breast cancer, which is the subject of the present invention. Moreover, it was a report of carcinogenesis in males. Therefore, there have been no examples that have verified the development of breast cancer by fluorinated estrogens.

Grodstein, F., Stampfer, M. J., Colditz, G. A., Willett, W. C., Manson, J. E., Joffe, M., Rosner, B., Fuchs, C., Hankinson, S. E., Hunter, D.J., Hennekens, C. H., and Speizer , F. E. Postmenopausal hormone therapy and mortality. N. Engl. J. Med. 336, 1769-1775 (1997). Colditz, G. A., Hankinson, S. E., Hunter, D. J., Willett, W. C., Manson, J. E., Stampfer, M. J., Hennekens, C. H., Rosner, B., and Speizer, F. E. The use of estrogens and progestins and the risk of breast cancer in Postmenopausal women. N. Engl. J. Med. 332, 1589-1593 (1995). Chen, C. L., Weiss, N. S., Newcomb, P., Barlow, W., and White, E. Hormone replacement therapy in relation to breast cancer. JAMA 287, 734-741 (2002). Beral, V. Million Women Study Collaborators. Breast cancer and hormone-replacement therapy in the Million Women Study. Lancet 362, 419-427 (2003). Lacey, J. V. Jr., Mink, P. J., Lubin, J. H., Sherman, M. E., Troisi, R., Hartge, P., Schatzkin, A., and Schairer, C. Menopausal hormone replacement therapy and risk of ovarian cancer. JAMA 288 , 334-341 (2002). Grady, D., Gebretsadik, T., Kerlikowske, K., Emster, V., and Petitti, D. Hormone replacement therapy and endometrial cancer risk: a meta-analysis. Obstet. Gynecol. 85, 304-313 (1995) . Bolton, J. L., Pisha, E., Zhang, F., and Qiu, S. Role of quinoids in estrogen carcinogenesis. Chem. Res. Toxicol. 11, 1113-1127 (1998). Han, X., and Liehr, J. G. 8-Hydroxylation of guanine bases in kidney and liver DNA of hamsters treated with estradiol: role of free radicals in estrogen-induced carcinogenesis. Cancer Res. 54, 5515-5517 (1994). Stack, D. E., Byun, J., Gross, M. L., Rogan, E. G., and Cavalieri, E. L. Molecular characteristics of catechol estrogen quinone in reactions with deoxyribonucleosides. Chem. Res. Toxicol. 9, 851-859 (1996). Shibutani, S., Takeshita, M., and Grollman, A.P. Insertion of specific bases during DNA synthesis past the oxidation-damaged base 8-oxodG. Nature349, 431-434 (1991). Terashima, I., Suzuki, N., and Shibutani, S. Mutagenic specificity of 2-hydroxyestrogen quinone-derived DNA adducts in mammalian cells. Biochemistry 40, 166-172 (2001). Suzuki, N., Itoh, S., Poon, K., Masutani, C., Hanaoka, F., Ohmori, H., Yoshizawa, I., and Shibutani, S. Translesion synthesis past estrogen-derived DNA adducts by human DNA polymerases η and κ. Biochemistry 43, 6304-6311 (2004). Malins, D. C., Holmes, E. H., Polissar, N. L., and Gunselman, S. J. The etiology of breast cancer. Characteristic alteration in hydroxyl radical-induced DNA base lesions during oncogenesis with potential for evaluating incidence risk. Cancer 71, 3036-3043 (1993) . Turan, V. K., Sanchez, R. I., Li, J. J., Li, S. A., Reuhl, K. R., Thomas, P. E., Conney, A. H., Gallo, M. A., Kauffman, F. C., and Mesia-Vela, S. The effects of steroidal estrogens in ACI Rat mammary carcinogenesis: 17β-estradiol, 2-hydroxyestradiol, 4-hydroxyestradiol, 16α-hydroxyestradiol, and 4-hydroxyestrone. J. Endocrinol. 183, 91-99 (2004). Shull, J. D., Spady, T. J., Snyder, M. C., Johansson, S. L., and Pennington, K. L. Ovary-intact but not ovariectomized female ACI rats treated with 17 β-estradiol rapidly develop mammary carcinoma. Carcinogenesis18, 1595-1601 (1997). Okamoto, Y., Liu, X., Suzuki, N., Okamoto, K., Kim, H. J., Y. R. Santosh Laxmi, Sayama, K., and Shibutani, S. Equine estrogen-induced mammary tumors in rats. 193, 224-228 (2010). Kirkman, H. Estrogen-induced tumors of the kidney. III. Growth characteristics in the Syrian hamster. Natl. Cancer Inst. Liehr, J. G. 2-Fluoroestradiol. Separation of estrogenicity from carcinogenicity. Mol. Pharmacol. 23, 278-281 (1983). Liehr, J. G., Stancel, G. M., Chorich, L. P., Bousfield, G. R., and Ulubelen, A. A. Hormonal carcinogenesis: separation of estrogenicity from carcinogenicity. Chem. Biol. Interact. 59, 173-184 (1986). Kuhl, H. "Pharmacology of estrogens and progestogens: influence of different routes of administration" (PDF). Climatic 8 (Suppl. 1), 3-63 (2005). Nakamura, H., Shiozawa, T., Terao, Y., Shiraishi, F., and Fukazawa, H. By-products produced by the reaction of estrogens with hypochlorous acids and their estrogen activities. J. Health Sci. 52, 124 -131 (2006). Hylarides, M.D., Leon, A. A., and Mettler, F. A. Synthesis of 1-chloroestradiol. Steroids, 43, 219-224 (1984). Mukawa, F. 10β-Chloro-17β-hydroxyestra-1,4-dien-3-one and its related compounds. J. Chem. Soc. Perkin Trans. 1, 457-459 (1988). Page, P.C.B., Hussain, F., Maggs, J.L., Morgan, P., and Park, B.K., Efficient regioselective A-ring functionalization of oestrogens. Tetrahedron, 46, 2059-2068 (1990). Y. R. Santosh Laxmi, Liu, X., Suzuki, N., Kim, S. Y., Okamoto, K., Kim, H. J., Zhang, G., Chen, J. J., Okamoto, Y., and Shibutani, S. Anti-breast cancer potential of SS1020, a novel antiestrogen lacking estrogenic and genotoxic actions. Int. J. Cancer 127, 1718-1726 (2010). Dunning, W.F., Curtis, M.R., and Segaloff, A. Strain differences in response to estrone and the induction of mammary gland, adrenal, and bladder cancer in rats. Cancer Res. 13, 147-152 (1953).

An object of the present invention is to provide an estrogen derivative that retains estrogen activity while suppressing carcinogenicity, thereby providing an estrogen preparation for estrogen replacement therapy.

As a result of intensive studies in order to solve the above problems, the present inventors found that estradiol derivatives having a halogen atom introduced at the 1-, 2- or 4-position have no carcinogenicity and retain estrogenic activity. , have completed the present invention.

（式中、Ｒ_１は、水素原子、炭素数１から６のアルキル基、炭素数２から７のアルキルカルボニル基、炭素数７から２０のアリールアルキル基、炭素数７から２０のアリールカルボニル基、PO₃H₂、又はＳＯ_３Ｎａを示し；
Ｒ_２は、水素原子、炭素数２から７のアルキルカルボニル基、炭素数７から２０のアリールカルボニル基、PO₃H₂、又はＳＯ_３Ｎａを示し、Ｒ_３は、水素原子、炭素数２から６のアルキレン基、又はＳＯ_３Ｎａを示し、Ｒ_４は、水素原子又はＯＨを示し、あるいは－ＯＲ_２と－Ｒ_３は一緒になって＝Ｏを示し；
Ｘ_１、Ｘ_２及びＸ_３はそれぞれ独立に、水素原子又はハロゲン原子を示し、但し、Ｘ_１、Ｘ_２及びＸ_３のうちの少なくとも一つはハロゲン原子である。）
［２］ Ｒ_１が、水素原子、ＣＨ_３、ＣＨ_３ＣＨ_２、ＣＨ_３ＣＯ、ＣＨ_３ＣＨ_２ＣＯ、ＣＨ_３（ＣＨ_２）_３ＣＯ、Ｃ_６Ｈ_５ＣＨ_２、Ｃ_６Ｈ_５ＣＯ、-PO₃H₂、又は－ＳＯ_３Ｎａである、［１］に記載のエストロゲン製剤。
［３］ Ｒ_２が、水素原子、ＣＨ_３、ＣＨ_３ＣＯ、Ｃ_６Ｈ_５ＣＯ、-PO₃H₂、又は－ＳＯ_３Ｎａである、［１］又は［２］に記載のエストロゲン製剤。
［４］ Ｒ_３が、水素原子、ＣＨ≡Ｃ、ＣＨ_３Ｃ≡Ｃ、又は－ＳＯ_３Ｎａである、［１］から［３］の何れか一に記載のエストロゲン製剤。
［５］ －ＯＲ_２と－Ｒ_３が一緒になって＝Ｏを示す、［１］又は［２］に記載のエストロゲン製剤。
［６］ Ｘ_１、Ｘ_２又はＸ_３が表すハロゲン原子が、Ｆ又はＣｌである、［１］から［５］の何れか一に記載のエストロゲン製剤。
［７］ Ｒ_１が水素原子であり、Ｒ_２が水素原子であり、Ｒ_３が、水素原子、又はＣＨ≡Ｃであり、Ｒ_４が水素原子であり、Ｘ_１、Ｘ_２及びＸ_３のうちの何れか一つがＦ又はＣｌであり、Ｘ_１、Ｘ_２及びＸ_３のうちの残りの二つが水素原子である、［１］に記載のエストロゲン製剤。
［８］ Ｒ_１が水素原子であり、－ＯＲ_２と－Ｒ_３が一緒になって＝Ｏを示し、Ｒ_４が水素原子であり、Ｘ_１、Ｘ_２及びＸ_３のうちの何れか一つがＦ又はＣｌであり、Ｘ_１、Ｘ_２及びＸ_３のうちの残りの二つが水素原子である、［１］に記載のエストロゲン製剤。 That is, according to the present invention, the following inventions are provided.
[1] An estrogen preparation for estrogen replacement therapy, containing a compound represented by the following formula (1).

(wherein R ₁ is a hydrogen atom, an alkyl group having 1 to 6 carbon atoms, an alkylcarbonyl group having 2 to 7 carbon atoms, an arylalkyl group having 7 to 20 carbon atoms, an arylcarbonyl group having 7 to 20 carbon atoms, represents _PO3H2 or _{SO3Na ;}
R ₂ represents a hydrogen atom _, an alkylcarbonyl group having 2 to 7 carbon atoms, an arylcarbonyl group having 7 to 20 carbon atoms, PO ₃ H ₂ , or SO ₃ Na; an alkylene group of _{6, or SO Na, R 4 represents} a hydrogen atom or OH, or -OR ₂ and -R ₃ together represent =O;
X ₁ , X ₂ and X ₃ each independently represent a hydrogen atom or a halogen atom, provided that at least one of X ₁ , X ₂ and X ₃ is a halogen atom. )
[2] R ₁ is a hydrogen atom, CH ₃ , CH ₃ CH ₂ , CH ₃ CO, CH ₃ CH ₂ CO, CH ₃ (CH ₂ ) ₃ CO, C ₆ H ₅ CH ₂ , C ₆ H ₅ CO, The estrogen preparation according to [1], which is -PO ₃ H ₂ or -SO ₃ Na.
[3] The estrogen formulation of [1] or [2], wherein R ₂ is a hydrogen atom, CH ₃ , CH ₃ CO, C ₆ H ₅ CO, —PO ₃ H ₂ , or —SO ₃ Na.
[4] The estrogen preparation according to any one of [1] to [3], wherein R ₃ is a hydrogen atom, CH≡C, CH ₃ C≡C, or —SO ₃ Na.
[5] The estrogen formulation of [1] or [2], wherein -OR ₂ and -R ₃ together represent =O.
[6] The estrogen preparation according to any one of [1] to [5], wherein the halogen atom represented by X ₁ , X ₂ or X ₃ is F or Cl.
[7] R ₁ is a hydrogen atom, R ₂ is a hydrogen atom, R ₃ is a hydrogen atom, or CH≡C, R ₄ is a hydrogen atom, and X ₁ , X ₂ and X ₃ The estrogen preparation according to [1], wherein any one of which is F or Cl and the remaining two of X ₁ , X ₂ and X ₃ are hydrogen atoms.
[8] R ₁ is a hydrogen atom, —OR ₂ and —R ₃ together represent ═O, R ₄ is a hydrogen atom, and any one of X ₁ , X ₂ and X ₃ one is F or Cl and the remaining two of X ₁ , X ₂ and X ₃ are hydrogen atoms.

INDUSTRIAL APPLICABILITY According to the present invention, there is provided a halogenated estrogen derivative that retains estrogenic activity while suppressing carcinogenicity. INDUSTRIAL APPLICABILITY According to the present invention, it is possible to provide an estrogen preparation for estrogen replacement therapy that is highly safe and has no carcinogenicity for breast cancer.

FIG. 1 shows the mechanism of DNA damage by human estrogen. FIG. 2 shows the structures of estrogens and chlorinated estrogens. X represents a hydrogen atom (E2 derivative) or an ethynyl group _{( EE2} derivative). Figure ₃ shows the results of the uterotrophic assay of E2 and its halogenated derivatives in ovariectomized rats. *p<0.05, **p<0.01, ***p<0.0001 (vs control group)

The present invention will be described in more detail below.
In the present invention, a highly safe estrogen derivative is provided by inactivating a specific site (FIG. 1: arrow site) of the chemical structure involved in the carcinogenic action of estrogen by a synthetic chemical method to eliminate DNA damage. succeeded in That is, in the present invention, a bulky and highly electrophilic halogen atom is introduced at the 1-, 2- or 4-position of estrogen.

In the examples below, 1-chloro-estradiol (1-ClE ₂ ), 2-chloro-estradiol (2-ClE ₂ ), 4-chloro-estradiol (4-ClE ₂ ), and 17 - Synthesize 2-chloro-17-ethinyl-estradiol (2-ClEE ₂ ) and 4-chloro-17-ethinyl-estradiol (4-ClEE ₂ ), which are ethynyl estrogens (Non-Patent Document 20), and put them into ACI rats As a result of follow-up observation for one year after administration, it was confirmed that none of the compounds had carcinogenicity. We also confirmed that chlorinated estrogens have strong estrogenic activity. Chlorinated estrogens have been detected in trace amounts as by-products of chlorination at sewage treatment plants (Non-Patent Document 21), and their chemical synthesis methods have also been published (Non-Patent Documents 22 and 23). Chlorinated estrogens have been detected as by-products, and there are no reports that chlorinated estrogens have estrogenic activity in animals. In the present invention, chlorinated estrogens were found to have estrogenic activity and, unlike estradiol (E2), to be non _- carcinogenic. Similarly, when fluorinated estrogens were also examined, it was found that 2-fluoro-estradiol (2-FE ₂ ) has no carcinogenicity in breast cancer and has estrogenic activity equivalent to that of E ₂ . It has been shown to be one of the halogenated estrogens that can be used in replacement therapy.

Estrogen preparations for estrogen replacement therapy according to the present invention include compounds represented by the following formula (1).

R ₁ is a hydrogen atom, an alkyl group having 1 to 6 carbon atoms, an alkylcarbonyl group having 2 to 7 carbon atoms, an arylalkyl group having 7 to 20 carbon atoms, an arylcarbonyl group having 7 to 20 carbon atoms, or SO ₃ Na indicates
Preferably, R ₁ is a hydrogen _atom , _CH3 , _CH3CH2 , _CH3CO , _CH3CH2CO , _{CH3 ( CH2 ) 3CO , C6H5CH2 , C6H5CO} , -PO ₃ H ₂ or -SO ₃ Na.
More preferably, R ₁ is a hydrogen atom.

_R2 represents a hydrogen atom, an alkylcarbonyl group having 2 to 7 carbon atoms, an arylcarbonyl group having 7 to ₂₀ carbon atoms, _-PO3H2 , or _SO3Na .
Preferably, R ₂ is a hydrogen atom, CH ₃ , CH ₃ CO, C ₆ H ₅ CO, -PO ₃ H ₂ or -SO ₃ Na.
More preferably, _R2 is a hydrogen atom.

R ₃ represents a hydrogen atom, an alkylene group having 2 to 6 carbon atoms, or SO ₃ Na.
Preferably, R ₃ is a hydrogen atom, CH≡C, CH ₃ C≡C, or —SO ₃ Na.
More preferably _, R3 is a hydrogen atom or CH≡C.

In the present invention, —OR ₂ and —R ₃ may together represent ═O. In this case, the compound represented by Formula (1) is a compound represented by Formula (1A) below. In the formula, the definition of R ₁ , R ₄ , X ₁ , X ₂ or X ₃ is the same as in formula (1).

_R4 represents a hydrogen atom or OH.
Preferably _R4 is a hydrogen atom.

X ₁ , X ₂ and X ₃ each independently represent a hydrogen atom or a halogen atom, provided that at least one of X ₁ , X ₂ and X ₃ is a halogen atom.
A halogen atom is preferably F, Cl, Br or I.
Preferably, the halogen atom represented by X ₁ , X ₂ or X ₃ is F or Cl.
More preferably, any one of X ₁ , X ₂ and X ₃ is F or Cl, and the remaining two of X ₁ , X ₂ and X ₃ are hydrogen atoms.

Synthesis of the compound represented by formula (1) includes, for example, Non-Patent Document 22 (1-ClE ₂ ), Non-Patent Document 23 (2-ClE ₂ , 4-ClE ₂ , 2-ClEE ₂ , 4-ClEE ₂ ), and Non-Patent Document 24 (2-FE ₂ , 4-FE ₂ ). Further, the synthesis of the compound represented by formula (1A) includes, for example, Non-Patent Document 23 (2-ClE ₁ , 4-ClE ₁ ), Non-Patent Document 24 (2-FE ₁ , 4-FE ₁ ), etc. It can be performed according to the method described in.

Estrogen is also called follicular hormone or female sex hormone, and is a kind of steroid hormone produced from cholesterol in ovarian granulosa cells, outer thecal cells, placenta, adrenal cortex, and testicular interstitial cells.

Known physiological functions of estrogen include promotion of mammary gland cell growth, control of ovarian ovulation, control of lipid metabolism, insulin action, blood coagulation action, feminization of the central nervous system (consciousness), thinning of the skin, and inhibition of arteriosclerosis. Estrogen includes estrone (E ₁ ), estradiol (17β-estradiol: E ₂ ), and estriol (E ₃ ), and estradiol (E ₂ ) has the highest activity.

Estrogen replacement therapy is a treatment method using estrogen or drugs that promote or inhibit the secretion or action of estrogen for diseases or symptoms caused or induced by estrogen deficiency.
For example, in order to alleviate menopausal disorders caused by a sudden decline in female hormones during menopause and to prevent osteoporosis caused by hormone decline, treatment can be performed by supplementing the deficient estrogen or administering other estrogen-activating chemical substances. be.

The compound represented by formula (1) can be used as a pharmaceutical composition, supplement or food for estrogen replacement therapy by preparing a formulation according to the route of administration together with a usual pharmacologically acceptable carrier. .

Pharmaceutical compositions or supplements for oral administration can be prepared, for example, in the form of tablets, capsules, granules, powders, liquids, and the like. When preparing solid preparations for oral administration, conventional excipients, binders, disintegrants, lubricants, wetting agents, emulsifiers, preservatives, sweetening agents, flavoring agents, coloring agents and the like can be used. can

Formulations for oral administration generally contain 0.01 to 100% by weight, preferably 0.05 to 100% by weight of the compound represented by formula (1) as an active ingredient.

Furthermore, the formulation of the present invention can be prepared by adding pharmacologically acceptable carriers, pH adjusters, buffers, stabilizers, isotonic agents, local anesthetics, etc. to form liquid formulations (injections, suspensions, Syrups, etc.), suppositories, patches, etc., for parenteral administration.

Liquid formulations are prepared by using appropriate additives commonly used in liquid formulations, such as water, alcohols, or plant-derived oils such as soybean oil, peanut oil, or sesame oil, and are in the form of suspensions, syrups, injections, or the like. can be manufactured as In particular, suitable solvents for parenteral administration by intramuscular injection, intravenous injection or subcutaneous injection include, for example, distilled water for injection, lidocaine hydrochloride aqueous solution (for intramuscular injection), physiological saline, glucose aqueous solution, Ethanol, liquids for intravenous injection (for example, aqueous solutions of citric acid and sodium citrate, etc.), electrolyte solutions (for intravenous drip and intravenous injection), and mixed solutions thereof can be mentioned.
Formulations for parenteral administration generally contain 0.01 to 10% by weight, preferably 0.05 to 5% by weight of the compound represented by formula (1) as an active ingredient.

Furthermore, the formulation of the present invention may be provided as cosmetics or external skin preparations (drugs, quasi-drugs). In cosmetics and external preparations for skin, the dosage form is not particularly limited as long as it is applicable to the skin, and examples thereof include liquids, creams, ointments, plasters, milky lotions, lotions, packs, and the like.

The dosage varies depending on the patient's symptoms, body weight, age, etc., but the amount of the compound represented by formula (1) is preferably in the range of about 0.1 to 100 mg per day, preferably about 0.1 mg per day. A range of -50 mg is more preferred, and a range of about 0.1 to 25 mg is more preferred, which is usually desirably administered in 1 to 4 divided doses per day.

The estrogen preparation of the present invention containing the compound represented by formula (1) as an active ingredient is an estrogen-like pharmaceutical composition with reduced side effects, for example, improving menopausal disorders and urogenital disorders such as atrophic vaginitis. , for the prevention or treatment of diseases such as amenorrhea, uterine hypoplasia, ovarian deficiency, prostate cancer, prostatic hyperplasia, osteoporosis, arteriosclerosis, memory impairment, Alzheimer's disease, thrombotic disease, etc. caused by decreased estrogen can be used for In particular, it can be used in place of estrogen in hormone replacement therapy for improving menopausal disorders. The estrogen formulation of the present invention is also effective as a health food or supplement having estrogenic activity.

According to the present invention, there is provided estrogen replacement therapy comprising administering a compound represented by formula (1) to a subject in need of estrogen.
According to the present invention there is provided a compound of formula (1) for use in estrogen replacement therapy.
According to the present invention there is provided use of a compound of formula (1) for the manufacture of estrogen preparations for estrogen replacement therapy.

The present invention will be described in more detail with reference to the following examples, but the present invention is not limited by the examples.

<Example 1>
The structures of the compounds used in Example 1 are given below.

(1) Synthesis of halogenated estrogens 1-ClE ₂ (Non-Patent Document 22), 2-ClE ₂ , 4-ClE ₂ , 2-ClEE ₂ , 4-ClEE ₂ (Non-Patent Document 23), and 2-FE ₂ , 4-FE ₂ (Non-Patent Document 24) were chemically synthesized by a previously reported method.

(2) Detection of Carcinogenicity of Halogenated Estrogen Pellets containing 2.5 mg or 5.0 mg of 1-ClE ₂ , 2-ClE ₂ and 4-ClE ₂ were prepared. Pellets containing 1.25 mg or _2.5 mg of E2 were prepared as positive controls, and pellets containing no compound were prepared as negative controls. Each pellet was subcutaneously inserted into 5-week-old female ACI rats and the progress was observed (Non-Patent Document 16). Table 1 shows the results of measuring the development of breast cancer. E ₂ showed 80% breast cancer development after several months. In contrast, neither 2.5 mg nor 5.0 mg of 1-ClE ₂ , 2-ClE ₂ and 4-ClE ₂ did not develop breast cancer even after one year of administration, as in the negative control. . In addition, no breast cancer occurred when 2-ClEE ₂ or 4-ClEE ₂ was administered at 2.5 mg or 5.0 mg. Furthermore, no carcinogenesis of uterine cancer and ovarian cancer was observed. 2-FE ₂ was also not found to be carcinogenic. From the above, it was confirmed that halogenated estrogens do not have carcinogenicity.

(3) Measurement of Estrogenic Activity of Halogenated Estrogens The estrogenic activity of chlorinated estrogens (1-ClE2, ₂ - _ClE2 , 4- _ClE2 ) was measured by subcutaneous injection of trace amounts into ovariectomized Sprague-Dawley rats. Three days after the administration of (3.4 μg), the uterus was excised and detected by calculating the uterine weight per rat body weight (Non-Patent Document 25) (Fig. 3). E ₂ (3.0 μg) in the same molar amount was administered as a positive control for this uterine hypertrophy, and saline alone was administered as a negative control. Strong estrogenic activity could be detected for each of the chlorinated estrogens, but it was weaker than that of the same molar amount of E ₂ . However, as seen in the examples of 1-ClE ₂ (0.34, 3.4, 34 μg), the estrogenic activity increased accordingly as the dose increased, and an estrogenic activity equivalent to that of E ₂ was obtained. Subcutaneous administration of 2-FE ₂ or 4-FE ₂ (3.2 µg) also showed the same uterotrophic effect as that of E ₂ (3.0 µg) of the same molarity (Fig. 3). It was confirmed that the result of the uterine enlargement effect of fluorinated estrogen obtained by the inventor of the present invention is equivalent to the already published report (Non-Patent Document 18). Therefore, chlorinated estrogens have strong estrogenic activity in animals, although _they are not as strong as E2, and fluorinated estrogens have estrogenic activity equivalent to E2, so _they can be used as hormone replacement therapy agents. is.

<Example 2>
The structures of the compounds used in Example 2 are given below.

As examples of compounds represented by formula (1A), 2-chloro-esrone (2-ClE ₁ ), 4-chloro-esrone (4-ClE ₁ ), or 2-fluoro-esrone (2-FE ₁ ) was synthesized. , pellets containing 2.5 mg were made. Pellets containing 2.5 mg or ₁₀ mg of E1 were prepared as positive controls, and pellets containing no compound were prepared as negative controls. Each pellet was subcutaneously inserted into 5-week-old female ACI rats, and the progress was observed. Table 2 shows the incidence of breast cancer for one year after administration. With E ₁ (2.5 mg), 1 out of 6 animals (17%) developed breast cancer. Compared to E ₂ at the same dose (Table 1), the lower incidence of breast cancer with E ₁ was consistent with reports by other research groups (Non-Patent Document 26). When the dose of E1 was increased to ₁₀ mg, 100% breast cancer incidence was confirmed 24 weeks after administration. In contrast, administration of 2.5 mg of 2-ClE ₁ or 4-ClE ₁ did not cause breast cancer even 1 year after administration, as in the negative control. 2-FE ₁ was also not carcinogenic. From the above, it was confirmed that halogenated estrone does not have carcinogenicity.

Claims (1)

An estrogen preparation for suppressing breast cancer carcinogenesis and estrogen replacement therapy, comprising a compound represented by the following formula (1).

(Wherein, R ₁ represents a hydrogen atom;
R ₂ represents a hydrogen atom,
R ₃ represents a hydrogen atom or CH≡C ,
R ₄ represents a hydrogen atom,
Any one of X ₁ , X ₂ and X ₃ is Cl, and the remaining two of X ₁ , X ₂ and X ₃ are hydrogen atoms.

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