TWI293251B - Selective estrogen receptor modulators incombination with estrogens - Google Patents

Description

1293251 V. DESCRIPTION OF THE INVENTION (1) Technical Jaws of the Invention The present invention relates to novel combinations of several physiologically active compounds. In particular, it includes a combination of a selective estrogen receptor modulator (SERM) and an estrogen. In some embodiments, the combination includes a selective estrogen receptor modulator (SERM), an estrogen, and a sex steroid or male compound precursor. The invention also provides several kits and pharmaceutical compositions for carrying out the foregoing combinations. Administration of the foregoing combination to a patient may reduce or eliminate the incidence of a tidal wave, vasomotor symptoms, dryness of the sheath, or other menopausal symptoms. The risk of developing breast cancer and/or endometrial cancer can be reduced after the patient receives the combination therapy. The case also provides a treatment or reduction of osteoporosis, biliary sclerotemia, hyperlipidemia, atherosclerosis, hypertension, Alzheimer's disease, insomnia, cardiovascular disease, insulin resistance, diabetes and obesity The method of possibility (especially obesity of the abdomen). BACKGROUND OF THE INVENTION A number of diseases, conditions and undesired symptoms are currently known to respond favorably to the administration of exogenous steroids or their precursors. For example, estrogen is thought to reduce the rate of bone loss, and males have been shown to build bone by stimulating bone formation. Hormone replacement therapy (eg, estrogen administration) is available for the treatment of menopausal symptoms. Luteinizing hormone is often used to combat the risk of endometrial hyperplasia and endometrial cancer caused by estrogen. The use of estrogens, androgen compounds and/or progesterone for a wide range of symptoms and conditions for the treatment or prevention of disease has created a number of disadvantages. Treatment of male compounds with males may produce some degree of masculine unwanted side effects. Similarly, vote for the patient --------------------- order --------- (please read the notes on the back and fill in the book) Page) Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printing

1293251 A7 B7 V. INSTRUCTIONS (Economics of the Intellectual Property Office of the Ministry of Economic Affairs. Employees' consumption of seroconversion steroids may increase the risk of certain diseases in patients. For example, breast cancer will be activated with estrogen. Prostate cancer and good; ^ The colonization will also deteriorate with the activation of androgen. Therefore, it is necessary to have a more effective and capable of reducing the risk of reducing the risk of sin. The combination therapy of the present invention and the medicinal composition of the therapeutic towel can be satisfied. These requirements. 概要 SUMMARY OF THE INVENTION The object of the present invention is to provide treatment or to reduce the risk of fever, renal vasomotor symptoms, osteoporosis, cardiovascular disease, hyperbilirubinemia, hyperlipidemia, arteriosclerosis A method of morbidity, hypertension, tamian resistance, diabetes, obesity (especially abdominal obesity), menstrual period (4), and the incidence or risk of saddle membrane dryness. Another purpose of this hair is to provide several treatments. Or reducing the risk of suffering from the above mentioned diseases while minimizing unwanted side effects. Another object of the present invention is to provide several suitable Kits and pharmaceutical compositions of the above methods. In one embodiment, the invention provides a method of reducing or eliminating the incidence of menopausal symptoms, the method comprising administering to the patient the aforementioned elimination or reduction of morbidity Administering to the patient a therapeutically effective amount of an estrogen or a prodrug thereof in combination with a therapeutically effective amount of a selective estrogen modulator or a prodrug thereof, the selective estrogen receptor modulator being different from estrogen Compound.

· ; Line · ' n n n (Please read the back note first and then fill out this page) --3⁄4 This paper size applies to China National Standard (CNS) A4 specification (210 X 297 public)

-6 1293251

The Ministry of Economic Affairs Intellectual Property Office employee consumption cooperative is printed in another specific example, and the present invention provides a method for treating or reducing the risk of suffering from a muddy condition selected from osteoporosis, hypercholesterolemia , hyperlipidemia, atherosclerosis, hypertension, Alzheimer's disease, insulin resistance, diabetes, sarcoplasmic loss, obesity, cerebral hemorrhage caused by hormonal therapy, and breast tenderness caused by hormonal therapy The method comprises the step of administering to the patient the aforementioned elimination or reduction of the morbidity, the therapeutically effective amount of an estrogen or a prodrug thereof, in combination with a therapeutically effective amount of a selective estrogen receptor modulator or a prodrug thereof The patient is administered a drug, and the selective estrogen receptor modulator is a compound different from estrogen. In another embodiment, the invention provides a pharmaceutical composition comprising: ', a) a pharmaceutically acceptable adjuvant, diluent or carrier; b) - a therapeutically effective amount of at least one estrogen or a prodrug thereof And c) a therapeutically effective amount of at least one selective estrogen receptor modulator or a prodrug thereof, wherein the selective estrogen receptor modulator is a compound different from estrogen. In another embodiment, the invention provides a kit comprising a first content comprising a pharmaceutical formulation, the first content comprising a therapeutically effective amount of at least one estrogen or a crude drug thereof, and the set The kit further comprises a second content comprising a pharmaceutical formulation comprising at least one therapeutically effective amount of a selective estrogen morphogen modulator or a prodrug thereof. In a specific embodiment, the present invention provides a method for treating or reducing the risk of osteoporosis in a patient, comprising administering an estrogen to the patient in a pharmaceutically effective manner, and further comprising administering a selectivity to the patient. The estrogen receptor regulates the paper scale and applies the Chinese National Standard (CNS) A4 specification (21〇X 297 public interest)" --------------Installation -------- Order ---------Line first read the back of the note and then fill out this page} 1293251

V. INSTRUCTIONS (4) The therapeutically effective amount of the agent (SERM) is part of a combination therapy. (Please read the note on the back and then fill out this page.) In another specific example, the present invention provides a method for treating or reducing the risk of cardiovascular disease, including the administration of H (tetracycline) to a patient. It further comprises administering to the patient a therapeutically effective amount of a selective estrogen receptor modulator (SERM) as part of a combination therapy. In another specific embodiment, the present invention provides a method for treating or reducing hyperbilirubinemia, comprising administering a pharmaceutically effective amount of estrogen to a patient and further including administering a patient to the patient. A therapeutically effective amount of a selective estrogen receptor modulator (SERM) is part of a combination therapy. In another embodiment, the present invention provides a method for treating or reducing the risk of suffering from serotonin, comprising administering to a patient a pharmaceutically effective amount of an estrogen, and further comprising administering a choice to the patient. A therapeutically effective amount of a sex estrogen receptor modulator (SERM) is part of a combination therapy. In another embodiment, the invention provides a method for treating or reducing the risk of suffering from atherosclerosis, comprising administering to the patient a pharmaceutically effective amount of an estrogen, and further comprising administering a choice to the patient A therapeutically effective amount of a sex estrogen receptor modulator (SERM) is part of a combination therapy. The Ministry of Economic Affairs' Intellectual Property Office employee consumption cooperation is printed in another specific example. 'The present invention proposes a method for treating or reducing the risk of suffering from blood pressure, including administering a pharmaceutically effective amount of estrogen to the patient' and It further comprises administering to the patient a therapeutically effective amount of a selective estrogen receptor modulator (SERM) as part of a combination therapy. In another embodiment, the invention provides a method for treating or reducing the risk of developing insomnia, comprising administering to the patient a pharmaceutically effective amount of an estrogen, and further comprising administering to the patient an optional female Hormone receptor modulator This paper scale applies to Chinese National Standard (CNS) A4 specification (210 X 297 mm) 1293251 A7 B7 V. Invention Description (SERM) is effective as part of a combination therapy. In a specific embodiment, the present invention provides a method for treating or reducing the risk of developing cognitive loss, comprising administering a pharmaceutically effective remedy to an estrogen, and further comprising administering a selective estrogen to the patient. A therapeutically effective amount of a receptor modulator (SERM) as part of a combination therapy. In another embodiment, the invention provides a method for treating or reducing the risk of developing Alzheimer's disease, including a disease A pharmaceutically effective amount of estrogen administered, and further comprising a therapeutically effective amount of a selective estrogen receptor modulator (SERM) administered to the patient as part of a combination therapy. The present invention provides a method for treating or reducing the risk of developing diabetes comprising administering a pharmaceutically effective amount of an estrogen to the patient, and further comprising administering to the patient a selective estrogen receptor modulator (SERM) The therapeutically effective amount is as part of a combination therapy. In another embodiment, the invention provides a method for treating or reducing the risk of developing menopausal symptoms, comprising administering an estrogen to the patient A pharmaceutically effective amount, and more particularly comprising administering to the patient a therapeutically effective amount of a selective estrogen receptor modulator (SERM) as part of a combination therapy. In another embodiment, the invention provides a A method of treating or reducing the risk of developing excessive obesity, particularly abdominal obesity, comprising administering to the patient a pharmaceutically effective amount of an estrogen, and further comprising administering to the patient a selective estrogen receptor modulator ( The therapeutically effective amount of SERM) is part of a combination therapy. In another embodiment, the invention provides a method for treating or reducing the risk of developing menopausal symptoms, including Cast a female ------------- loading -------- order --------- line (please read the back of the note before you fill out this page 9 A7 B7 1293251 V. INSTRUCTIONS (6) (Please read the note on the back and fill out this page) Pharmacologically effective amount' and more include a selective estrogen receptor modulator (SERM) for the patient Therapeutically effective amount is part of a combination therapy. In another embodiment, the invention provides a method for treating or reducing the risk of developing breast tenderness caused by hormonal therapy, comprising administering an estrogen to the patient A pharmaceutically effective amount, and more particularly comprising administering to the patient a therapeutically effective amount of a selective estrogen receptor modulator (SERM) as part of a combination therapy. In another embodiment, the invention provides a A method for treating or reducing the risk of developing a sheath-bleeding caused by hormonal therapy, comprising administering to the patient a pharmaceutically effective amount of an estrogen, and further comprising administering to the patient a selective estrogen receptor modulator (SERM) The therapeutically effective amount is part of a combination therapy. i-Line - Ministry of Economic Affairs Intellectual Property Office Staff Consumer Cooperative Printed in another specific example, the present invention proposes a method for treating or reducing the incidence of osteoporosis by increasing the level of a monos-steroid precursor, the sex steroid The precursor is selected from the group consisting of dehydroepiandrosterone (Dhea), dehydroepiandrosterone-sulfate (DHEA-S), androstenedione and androst-5-ene 3,17 diol (5) - a group of diols, which is used in the above-mentioned patients in need of treatment or risk reduction, and more includes a therapeutically effective amount of a selective estrogen receptor modulator (SERM) administered to the patient. A therapeutically effective amount with an estrogen as part of a combination therapy. In another embodiment, the present invention provides a method for treating or reducing the incidence of orgasm or sweating by increasing the level of a monos-steroid precursor selected from the dehydrogenation table. Male Fat Liao (DHEA), Dehydroepiandrosterone-Sulphate (DHEA-S), Androstene Di- and Male-Olefin _% Alkene 3 This paper scale applies to China National Standard (CNS) A4 specification (21Q x 297) Love) ' ---- -10 - Ministry of Economic Affairs, Intellectual Property Bureau, employee consumption cooperative, printing

1293251 A7 —----- B7________ V. Group of Inventions (7) / 3,17 diol (5-diol), which is used in the above-mentioned patients who need treatment or reduce risk And further comprising administering to the patient a therapeutically effective amount of a selective estrogen receptor modulator (SERM) and a therapeutically effective amount of an estrogen as part of a combination therapy. In another embodiment, the invention provides a method of treating or reducing the risk of suffering from such diseases as mentioned above, comprising administering to the patient a therapeutically effective amount of a conjugate/antagonist estrogen (mixed SERM) And further comprising administering to the patient a therapeutically effective amount of a pure selective estrogen receptor modulator (pure SERM) 2 and a therapeutically effective amount of an estrogen as part of a combination therapy. As used herein, ''mixed SERM') means that the SERM has some estrogen activity in the breast and endometrial tissue at a physiological or pharmacological concentration, as used herein, 'pure SERM' means SERM in the breast and uterus In membrane tissue, the concentration does not have estrogenic activity at physiological or pharmacological concentrations. In another embodiment, the present invention provides a kit comprising a first content, the first content comprising at least a therapeutically effective amount of estrogen, and further comprising - a second content, The content contains a therapeutically effective amount of at least one selective estrogen receptor modulator. In another embodiment, the invention provides a pharmaceutical composition comprising: a) a pharmaceutically acceptable adjuvant, diluent or carrier; b) at least one therapeutically effective amount of at least one estrogen '· and c) at least A therapeutically effective amount of a selective estrogen receptor modulator. As used herein, 'combined, other compounds that are administered to patients. This paper scales the National Standards (CNS) A4 specification 5ί〇X 297 public i -------------- ---^ C Please read the notes on the back and fill out this page.} 11 A7 1293251 V. Inventive Note (8) is administered at a dose close enough to the other compound, so the patient can simultaneously obtain the physiology of the two compounds. The reaction, even if the compounds are not administered in a similar period of time, when the compounds are administered as part of a combination therapy, it means that the compounds are administered in combination with each other. Estrogen replacement therapy is often followed by menopause. Women are used to avoid and treat diseases caused by menopause, namely osteoporosis, craze, coronary heart disease (Cummings 1991), but there are some undesired effects associated with chronic estrogen administration. The increased risk of detectable uterine cancer and/or breast cancer produced by estrogen (Judd, Meldrum et al. 198; Colditz, Hankinson et al. 1995) is the greatest drawback of this treatment. The authors of the present invention found that The addition of a selective estrogen receptor modulator (SERM) during hormone administration inhibits such undesired effects. The present invention provides a method for treating or reducing the risk of breast tenderness caused by hormone replacement therapy (HRT). The method, because SERM can cause breast epithelial atrophy 'instead of the stimulation caused by HRT, so breast tenderness can be slowed or eliminated. The present invention also provides a method for avoiding and treating hormonal replacement therapy leading to sheath bleeding. Because SERM can cause The atrophy of the endometrium will not occur in the sheath. On the other hand, the use of SERM alone does not have a significant or ineffective effect on certain menopausal symptoms such as craze and sweating. It is believed that the addition of estrogen to SERM therapy for the treatment of menopausal symptoms can reduce or even eliminate the excitement and sweating. It is important to note that the craze and sweating are the first signs of menopause and that the patient accepts or Do not accept the menstrual wood paper scale applicable to the Chinese National Standard (CNS) A4 specification (21〇x 297 mm (please read the back of the note first and then fill out this page) Order --- Line - Ministry of Economic Affairs Intellectual Property Bureau Staff Consumer Cooperative Printed 12 A7 1293251 ___B7 ___ V. Invention Description (9) Treatment often depends on success or unsuccessful reduction of fever and sweating. As used here, A selective estrogen receptor modulator (SERM) is a compound that acts as an estrogen receptor antagonist ("anti-estrogen") in breast tissue either directly or via its activated metabolite, however it is for bone Tissue and serum cholesterol levels produce estrogen or estrogen-like effects (ie, lower serum cholesterol levels). It is possible to use a SERM in vitro or in a human or large-mouse breast tissue using a non-steroidal compound as an estrogen receptor antagonist (especially if the compound acts as an anti-estrogen against human breast cancer cells) effect. Conversely, steroid antiestrogens tend not to play the role of SERM because steroid antiestrogens tend to have no beneficial effects on serum cholesterol. Non-steroidal antiestrogens that we have tested and found to be used in SERM include EM-800, EM-652.HC1, Raloxifene, Tamoxifene, 4-carbyl-mosasfene (4- hydroxy-Tamoxifene), toremifene, 4-hydroxy-Toremifene, Droloxifene, LY 353 38, LY 335 563, GW-5638, Lasso-Foxifene Lasofoxifene), TSE 424 and Idoxifene, but are not limited to such compounds. However, we have also found that not all SERMs have the same function and can be divided into two sub-categories: “pure SERM” and “hybrid SERM”. Therefore, some of the physiological or pharmacological concentrations like SEMM of EM-800 and EM-652.HC1 do not have any estrogen activity in the breast and endometrial tissues, and have a low cholesterol and three in rats. The role of acid glycerides is too low. These SERMs can be referred to as "pure SERMs". The ideal paper size applies to the Chinese National Standard (CNS) A4 specification (210 X 297 mm) -------------Installation--------Set------- -- Line (please read the notes on the back and fill out this page) Printed by the Ministry of Economic Affairs, Intellectual Property Bureau, Staff and Consumer Cooperatives 13 A7 1293251 _ B7 _ V. Invention Description (10) (Please read the notes on the back and fill in the form) The SERM of the page is a pure SERM of the EM-652.HC1 type because of its potential and pure antiestrogenic activity in the mammary gland. Others, such as Raloxifene, Tamoxifene, Droloxifene, 4-carbyl-Tamoxifene (l_(4-didecylamine ethoxyphenyl)-1 -(4-phenylphenyl)-2-phenyl-butan-1-baked), multidimensional rice, 4-hydroxy-dolomifen [(Ζ)-(2)-2-[4(4-气- 1-(4-Hydroxyphenyl)-2-phenyl-1-butenyl)phenoxy]-indole, hydrazine dimethyl ethaneamine], LY 353 38 LY 335 563, GW-5638 and Idoxifene has some estrogenic activity in both the breast and the endometrium. This second series of SERMs can be referred to as "hybrid SERMs." The undesired estrogenic activity of such "mixed SERM" can be inhibited by the addition of a pure "SERM", such as the in vitro test shown in Figures 6 and 7, and the breast cancer shown in Figure 9 in vivo. Because human breast cancer xenografts in nude mice are the closest available model for human breast cancer, we compared EM-800 and its Tamoxifene alone and in combination with ZR-75-1 breast cancer xenografts in nude mice. Impact of transplant growth 〇 Ministry of Economic Affairs Intellectual Property Bureau Employees Consumption Cooperatives Printed for all combinations of teachings here, it is considered that each component of the combination is administered individually, except where otherwise specified. Thus, for example, administration of a SERM and an estrogen means administration of two different compounds rather than administration of a single SERM compound with some estrogenic properties. Applicants believe that the SERM of the present invention is in the breast, The role of pure antiestrogens in the tissues of the uterus and endometrium is very important, because SERM must fight against the potential estrogen to increase the risk of cancer in these tissues. The scale applies to China National Standard (CNS) A4 specification (210 X 297 mm) 14 1293251 A7 B7 Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printing skin agent swollen five, invention description (11 side effects. Applicants particularly believe that the present invention The benzofuran derivative has an absolute configuration of 2S in the second position and is therefore more suitable than its racemic mixture. Therefore, in US 6,060,503, a photoactivated scorpion anti-estrogen with a 2S configuration is disclosed Treatment of estrogen-induced breast and endometrial cancers' also shows that these compounds are more effective than their racemic mixtures (see Figures 1 to 5 of US, 060, 503). It is difficult to obtain pure state 2S in industry. For the configured mirror image, the Applicant believes that the '5R mirror image contaminant weight is less than 10%, preferably less than 5% and more preferably less than 5%. It is preferred that the figure 1 shows DHEA (10 mg, percutaneously, once per week) 800 (75 " g, orally, once daily) alone or in combination for 9 months in rat serum triglyceride (A) and cholesterol (B) The effect of the level. The data is represented by the mean SEM. **: Ρ<〇·〇1, the experimental group compared their respective control groups. Figure 2 shows: Α) In the ovariectomized (OVX) and female-supplemented nude mice, increasing the dose of DHEA (0.3 The effect of transdermal administration of mg, 1·〇mg or 3.0 mg) on the average ZR-75-1 tumor volume twice a day. Control group OVX mice receiving vehicle alone were used as an additional control group. The volume is taken as 100%. DHEA is administered percutaneously (pc) to the dorsal surface in a solution containing 50% ethanol and 50% propylene glycol in 0.02 ml. B) in ovariectomy (〇νχ) In nude mice supplemented with female supplements, the effect of increasing amounts of DHEA or EM-800 alone or in combination on the tumor weight was 9.5 months. ρ<0·01, accepting the processor relative to the control group of women--------------------- order-------- (please read first Precautions on the back side of this page) 15 1293251 A7 B7 V. Description of the invention (l2 supplementation of OVX mice for comparison. Figure 3 shows an increase in oral administration in ovarian resection (〇νχ) nude mice supplemented with female hormones. Dosage of anti-estrogen EM-800 (15 mg, 50 mg or 1 mg) (B) or increased dose of DHEA (0.3, 1.0 or 3.0 mg) transdermally with EM-800 (15 mg The effect of combination or EM-800 treatment (A) for 9.5 months on the mean ZR-75-1 tumor volume. The initial tumor volume was taken as ι〇〇0/〇. Control group OVX mice receiving vehicle alone It was used as an additional control group. The female system was administered once a day at a dose of 〇·5 " g, while DHEA was dissolved in 50% ethanol-50% propylene glycol twice daily. It was applied to the dorsal epidermal area in a volume of 0.02 ml, which was also compared with the OVX animals receiving the vehicle alone. The figure 4 was not found in the nematode-removed rats at a dose of gram per kg body weight; Mg anti-estrogen EM-800 treatment 65 days (mouth Serve once a day) or with methyl ethion progesterone acetate (MPA, lmg sc· 2 times daily) or combination EM-800 (0.25 mg/Kg body weight) and MPA for EJ1.04" g, Sc, 2 times a day) promotes the effects of DMBA on breast cancer growth. The change in tumor volume is expressed as a percentage of the initial tumor volume. The data are expressed as mean soil SEM. Figure 5 shows administration of oM-8(R) (〇·〇1, 0.03, 0·1, 0.3, and 1 mg/kg) or laroxine (Ral〇xifene) in ovariectomized rats 37 Week's effect on serum cholesterol levels. The line was compared with intact rats and 17/3-estradiol (EJ ovariectomized animals; ** ρ < 0 · (Η, experimental group vs. OVX control rats. Figure 6 shows in human I In Ishikawa cells, increase the concentration (please read the notes on the back and fill out this page) - Line · Ministry of Economic Affairs Intellectual Property Bureau Staff Consumer Cooperative Printed 16 - 1293251 A7 _ B7 V. Invention Description (13 ) -------------- Installed - (Please read the back of the urgent matter and then fill out this page) Degree of EM-800, (Ζ) · 4-0 Η - it Moxifen ( Tamoxifene), (Z)-4-0H-Toremifene and Raloxifene have effects on alkaline phosphatase activity. Alkaline ligase activity is exposed to increasing concentrations of the specified compound 5 days later at 1 · 0 The measurement is performed in the presence or absence of nM E2. The data is represented by the mean SEM of the four sample wells. When the SEM overlaps with the used symbols, only the symbols are indicated (Simard, Sanchez et al., 1997). The figure shows that in human Ischwara cancer cells, (z)-4-OH-Tamoxifene, (Z)-4-OH-Dolomide ( Toremifene), Droloxifene and Raloxifene block the action of stimulating alkaline phosphatase activity via antiestrogens EM-800. Alkaline phosphatase activity is 5 曰 after exposure to increasing concentrations of the specified compound. Measurements were performed in the presence or absence of 30 or 100 nM EM-800. Data were presented as SEM of the average of 8 sample wells, while data from the control group were obtained from 16 sample wells (Simard, Sanchez et al., 1997). - 丨 line _ Figure 8 shows the comparison of the effects of standard HRT (estrogen) and SERM (EM-652) on menopausal parameters. Adding SERM to standard HRT will offset the strong negative effects of estrogen. The 9th chart of the property bureau employee consumption cooperation Du printing shows that the stimulating effect of Tarn〇xifene on human breast cancer ZR-75-▲ 1 xenograft growth is completely blocked by EM-652.HC1, which is simultaneously administered. This result is consistent with the fact that the pure antiestrogenic activity of EM-652.HC1 itself has no effect on tumor growth in the presence of Tamoxifene. Figure 10 shows the section of the rat mammary gland. A. Untreated animals. Leaflet (L) from less Cogging formed. This paper inserted applicable China National Standard Scale (CNS) A4 size (210 X 297 mm) A7 1293251 B7__ ___ five described ([mu]) of the invention shows a high magnification photograph cogging. B. Animals treated with EM-800 (0.5 mg/kg, body weight/day) for 12 weeks. The leaflet (L) is reduced in size. The high magnification photo inserted shows the atrophic alveolar cells. Figure 11 shows a section of the rat endometrium. A. Untreated animals. The luminal epithelium (le) is characterized by its column epithelial cells, while the glandular epithelium (GE) is a relatively cubic cell. The matrix contains some cellular elements and collagen fibers. B. Animals treated with EM-800 (0.5 mg/kg, body weight/day) for 12 weeks. The lumen epithelial height is significantly reduced. Glandular epithelial cells have unstained cells without active markers. The matrix is highly honeycombed due to a decrease in intercellular matrix elements. Figure 12 shows increasing concentrations of EM652.HC1, Lassofoxifene (free base; activated and deactivated mirrors) and Raloxifene for oral administration of ovariectomized mice treated with female hormones 9 In the future, the effect on the weight of the uterus. *ρ<〇·〇5, Q1, compare the control group that processed E!. Figure 13 shows increasing concentrations of EM652.HC1, Lassofoxifene (free base; activated and deactivated mirrors) and Raloxifene for oral administration of ovariectomized mice treated with both females 9 After the sputum, the effect on the weight of the sheath. **ρ<0·01, for comparison with the control group. Figure 14 shows EM-652.HC1, lasofoxifene (free test; activation and deactivation of mirror image) and 杈, Rossi of l"g and ΙΟ/zg. This paper scale applies to Chinese national standards. (CNS) A4 specification (210 X 297 mm) (Please read the back of the note first and then fill out this page) · -1 line - Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printing 18 A7 1293251 ___B7__ V. Description of invention ( 15) (Raloxifene) Effect on uterine weight after oral administration of 9 曰 in ovariectomized mice. **ρ<0·01, compared to the OVX control group. (Please read the notes on the back and fill out this page.) Figure 15 shows £1^-652. of 1/^ and 10/^: «(:1, Lasofofoxifene (free test; activation) And deactivating the mirror image) and the effect of Raloxifene on the weight of the sheath after oral administration of 9 曰 in ovariectomized mice. **ρ<0·01, compared with the OVX control group. Displayed in OVX rats with defined osteogenic defects, with E2, EM-652.HC1, E2+EM-652.HC1, DHEA, DHEA+EM-652.HC1 and DHEA+EM-652.HC1+E2 Effects of lumbar spine BMD after 26 weeks of treatment. Intensive control and OVX control animals were included in the control group. Figure 17 is shown in E2, EM-652 in OVX rats with defined osteogenic defects. .HC1, E2+EM-652.HC1, DHEA, DHEA+EM-652.HC1 and DHEA+EM-652.HC1+E^ After 26 weeks of treatment, the effect on BMD of the femoral spine. Complete control group and OVX control group Animals were included in the control group. The 18th chart printed by the Ministry of Economic Affairs, Intellectual Property Office, and the Consumer Cooperatives, is shown in OVX rats with definite bone formation defects. E2, EM-652.HC1, E2 Effects of +EM-652.HC1, DHEA, t DHEA+EM-652.HC1 and DHEA+EM-652.HC1+E2 on total body fat after 26 weeks of treatment. Complete control and οβχ control animals included In the control group, Figure 19 shows the effect of antiestrogens on the growth of ZR-75-1 tumors. In ovariectomized nude mice, 7 antiestrogens were treated with 161 曰 for females to guide the growth of human ZR-75- 1 The effect of breast tumor. The tumor volume is applied to the Chinese National Standard (CNS) A4 specification (210 X 297 mm) on the initial swollen paper scale. 19 1293251 A/ ----- B7 Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative print V. INSTRUCTIONS (16) Percentage of tumor area (i曰=1〇0%). Data are expressed as mean soil SeM (n=18 to 30 tumors/group); ## ρ<〇.〇ι , for EM-652.HC1; ** ρ<0·01, for OVX. Anti-estrogen is administered once daily by female stimulating dose. The dose is 50 // g/mouse. Subcutaneous sputum. 5cm contains a 1:25 ratio of female and cholesterol soft plastic injection. Figure 19B shows the effect of anti-estrogen on ZR-75-1 tumor growth . In ovariectomized nude mice, female hormone influence on day 161 of the guide Growth ZR-75-1 human breast tumors within seven antiestrogens process. Tumor volume is expressed as a percentage of the initial tumor area (1st = 100%). Data are expressed as mean ± SEM (n = 18-30 tumors/group); ## ρ < 0·01 vs. EM-652.HC1; ** ρ < 0.01 vs. OVX. The antiestrogens were orally administered once daily without the stimulation of estrogen at a dose of 100# g/mouse. Figure 19C shows the effect of antiestrogens on the growth of ZR-75-1 tumors. In ovarian resected nude mice, 7 antiestrogens were treated for 161 days on the effects of female-induced growth of human ZR-75-1 breast tumors. Tumor volume is expressed as a percentage of the initial tumor area (Day 1 = 100%). Data are expressed as mean ± SEM (n = 18-30 tumors/group); ## ρ < 0·01, vs. EM-652.HC1; ** ρ < 0.01 ' vs. OVX. The antiestrogens were administered orally once daily without the stimulation of estrogen at a dose of 200/g/mouse. Figure 20A shows the effect of antiestrogens on the response range. The effect of seven antiestrogens administered on 161 days on the response of human ZR-75-1 breast tumors in ovarian excision in nude mice. Complete degeneration indicates that those tumors could not be measured at the end of the treatment; partial degradation corresponds to those tumors degenerated >50% of the original volume; stable response refers to tumor regression <50% or evolution of $50%, and (please read the back first) Note: Please fill out this page) - Set · · i line · This paper scale applies to China National Standard (CNS) A4 specification (210 X 297 mm) 20 1293251 A7 B7 Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative prints V. DESCRIPTION OF THE INVENTION (1? Evolution refers to an evolution of more than 50% when compared with the original volume of the tumor. The anti-estrogen is administered orally once a day under the stimulation of a female hormone at a dose of 5 〇 to ^ mice, which is stimulated subcutaneously. 〇 5cm contains a ratio of 丨: 25 female soft and cholesterol soft plastic injection. Figure 20B shows the effect of anti-estrogen on the response range. In ovarian resected nude mice, 7 anti-estrogen administration 161 days to human 211 -75-1 The effect of the response range of breast tumors. Complete degeneration indicates that those tumors cannot be measured at the end of the treatment; partial degradation corresponds to those tumors degraded by > It should be referred to as tumor regression <5〇% or evolution $5〇G/(), and evolution refers to evolution of more than 50% when compared to the original tumor volume. Anti-estrogen is administered orally once daily without estrogen stimulation The dose is 200 # g/mouse. Figure 20C shows the effect of antiestrogens on the response range. In the ovariectomized nude mice, the response range of 7 antiestrogens administered to human zR-75 - 丨 breast tumor in 161 days The effects of complete degradation indicate that those tumors cannot be measured at the end of the treatment; partial degradation corresponds to those tumors that degenerate >5〇% of the original volume; stable response refers to tumor regression <50% or evolution $5〇%, and Evolution refers to an evolution of more than 50% when compared to the original volume of the tumor. The anti-estrogen is administered orally once a day without the stimulation of estrogen at a dose of 2 g/mouse. Figure 21 shows daily oral administration. Effect of EM-652.HC1 treatment on uterine weight for 2 weeks in ovariectomized rats fed 17 stone-estradiol (2 mg/kg) 'increased dose range from 〇·〇1 mg/kg to 10 Mg/kg. Use intact animals as an additional control group. ---------- -----------Book.--I---II (Please read the notes on the back and fill out this page)

A7 1293251 B7_ V. INSTRUCTIONS (18) Figure 22 shows ER-652.HC1 treatment in the uterus for 2 weeks in ovariectomized rats fed orally with S; estradiol (2 mg/kg) The effect of the height of the membrane epithelium is increased from 〇.〇lmg/kg to 10 mg/kg. Whole animals were used as an additional control group. Figure 23. The stained sections of hematoxylin and eosin in rat uterus were shown from intact control group (A), OVX control group (B), OVX+ E2 (2 mg/kg) and (C) 0VX+E2+EM-652. Epithelial lining cells were obtained from rats treated with HC1 (3 mg/kg) for 14 days. The stimulation of estradiol on endometrial epithelial cells was restored by simultaneous administration of EM-652.HC1 (magnification: X 700). BM: base film. Figure 24 shows the effect of EM-652.HC1 treatment on the weight of the sheath for 2 weeks in ovariectomized rats fed orally with 17/5-estradiol (2 mg/kg) daily, increasing the dose range per dose. From 0.01 mg/kg to 10 mg/kg. Whole animals were used as an additional control group. Figure 25 shows the effect of EM-652.HC1 treatment on serum cholesterol for 2 weeks in ovariectomized rats fed orally with 17/3-estradiol (2 mg/kg), increasing the dose per dose from 0.01 mg/kg to 10 mg/kg. Whole animals were used as an additional control group. DETAILED DESCRIPTION OF THE INVENTION As can be seen from Figure 9, the effect of approximately 100% of Tamoxifene on tumor growth is completely blocked by EM-652.HC1 treated at the same time. Based on its pure antiestrogenic activity, EM-652.HC1 does not produce any effect in stimulating human breast cancer ZR-75_1 xenograft growth in nude mice (Fig. 9). This paper scale applies to China National Standard (CNS) A4 specification (210 X 297 mm) (please read the note on the back and fill out this page) Order --- Line - Ministry of Economic Affairs Intellectual Property Bureau Staff Consumer Cooperative Printed 22 1293251 A7 _______B7 V. INSTRUCTIONS (19) We also tested the anti-estrogen ICI 182,780 of steroids and found that it does not function as a SERM. In accordance with the present invention, SERMs can be administered at the same dosages as are well known in the art, and even the art uses them as antiestrogens rather than SERMs. We also noticed a relationship between SERM's beneficial effects on serum cholesterol and the effects of androgen or estrogen-like effects on bone. SERM also has beneficial effects on hypertension, insulin resistance, diabetes and obesity (especially abdominal obesity). Without wishing to be bound by theory (many SERMs preferably have 2 aromatic rings bonded by 1 or 2 carbon atoms), it is generally believed and expected that the SERMs will be molecularly identified by the aforementioned receptors. The part acts on the estrogen receptor. The preferred dipstick "has a side chain that selectively causes antagonism in the breast and general uterine tissue without causing significant antagonism in other tissues. Therefore, the SERM can be exerted in the breast as desired. Anti-estrogen function, and unexpectedly exerts estrogen function (or provides estrogen-like activity) in bones and blood (where lipid and cholesterol concentrations are beneficially affected). This is beneficial for cholesterol and lipids. The role of conversion to a beneficial effect against atherosclerosis, it is generally known that atherosclerosis can be adversely affected by improper cholesterol and lipids. Another aspect 'osteoporosis, hyperbiliary septicemia, hyperlipidemia, ~ De and arteriosclerosis advantageously responds to the activity of estrogen or estrogen. According to the invention, by using the combination of estrogen and SERM, the desired fruit can be achieved in the target tissue, * not in other Undesirable political consequences in the organization. For example, a combination of estrogen and SERM can be found in the bone network (or on --------------) (please read the back Precautions Fill in this page) Order: -1 line · 痤 郎 曰 曰 曰 讨 讨 11 11 11 11 11 consumption ^ 阼 ^ printing

1293251 A7 B7 V. INSTRUCTIONS (2〇) Lipids or cholesterol) produce a beneficial estrogenic effect, while avoiding undesired estrogen effects in the breast and uterus, as SERMs used as estrogen antagonists The effect of estrogen on the breast and endometrium is efficiently blocked, as shown in Figures 10 and 11. As disclosed in Figure 10, although the circulating level of 17 estradiol increased from 95.9 ± 32.4 § / 1111 in intact animals to 143.5 ± 7.8 § / 1111 (0.5 mg / kg EM per oral treatment) An increase of 50% in 800 12-week animals, but significant breast atrophy was still observed. Similarly, in the figure, significant endometrial retraction was observed in animals receiving ΕΜ-800 (〇·5 mg/kg). In these intact animals receiving pure antiestrogens, Em-800, the inhibitory effect of estrogen at the level of the hypothalamus and pituitary was removed, resulting in an increase in LH and a 17__ A secondary increase in estradiol. In the 6-month study of intact rats receiving 0.5 mg/kg of the same dose of em-652 per sputum, the Intellectual Property Office of the Department of Intellectual Property, the concentration of anti-estrogen EM_652 was measured at 04 ng/ in the circulation. Ml (URMA-05-011-94). Because the average serum EM_652 concentration was 7 3 ± 〇 77 ng/ml in women who received an oral dose of 2 〇 mg EM-800 daily, it is clear that postmenopausal female estrogen replacement therapy is not Will affect the strong inhibition of EM-652 and its ability to avoid breast and endometrial cancer. The first phase of the study showed that em_8〇〇 and em_652 hci almost obtained the results of quasi-accumulation of serum EM-652. The undesired effect is also reduced in a synergistic manner by the combination used in the present invention. For all diseases described here, any other effect on breast tissue caused by the replacement agent summer estrogen is efficiently applied by SER Μ This paper scale applies the Chinese National Standard (CNS) A4 specification (210 X 297 public love · 1293251

V. Description of the invention (After the Ministry of Intelligence, the Intellectual Property Officer X

Blocking the anti-estrogen effects of breast tissue, as shown in Figures 2 and 3. The same conclusion can be obtained from Figure 10. In a preferred embodiment, the sex steroid precursor (dehydroepiandrosterone, dehydroepian male _-sulfate, male _5 ene-3, 170 diol, renol) , 17-diketone and its original drug) or androgen reagents are added to provide a beneficial androgenic effect, particularly to reduce the risk of suffering from bone diseases or to treat bone diseases. The combination of SERM and estrogen reduces or even stops bone degradation in the treatment of osteoporosis. When male or DHEA (and other sex steroid precursors) are further added, damaged bone tissue can be reconstituted. The sex steroid precursor (for other treatments of hyperbilirubinemia, hyperlipidemia, menopausal symptoms, Alzheimer's disease, cardiovascular disease, breast cancer, uterine cancer and ovarian cancer) can be combined with SERM The combination with estrogen works synergistically to provide a better treatment for the above diseases. This synergy is caused by factors other than male hormones (or sex steroid precursor steroids metabolized to males in peripheral tissues), and estrogen or redundancy "acts through different mechanisms. In some specific examples Luteinizing hormone is added to provide further androgenic effects. As is well known in the art, progesterone can be used in low doses without adversely affecting receptors other than androgen receptors (e.g., corticosteroid receptors). Lutein hormones also contain no undesired male side effects (such as facial hair damage in female patients). Hot waves, cardiovascular symptoms, Alzheimer's disease, cognitive loss and insomnia are involved in the central nervous system. Specific estrogen receptors. Low levels of brain estrogen may explain at least some of these symptoms. Exogenous females

This paper scale applies to the country's national fresh (CNS) A4 specifications (2) Gx297 public love) ------------- loaded --------- set --------- Line f Please read the notes on the back and fill out this page. A7 1293251 V. INSTRUCTIONS (22) Hormones and specific estradiol can pass through the blood-brain barrier and bind to estrogen receptors to restore normal estrogen activity. On the other hand, the SERM of the present invention (more specifically, the EM-652.HC1 population) cannot pass through the blood-brain barrier, as shown in Example 9. Therefore, they are unable to antagonize the positive effects of estrogen in the brain, but they can antagonize the negative effects of estrogen on the breast, uterus and endometrial tissue. The combination (SERM + estrogen) is used separately for treatment or reduction. Special attention is paid to the risk of the above conditions. Total Addition Benefits of Estrogen and SERM Combinations The main reason for women looking for their physicians to discuss menopause is because of the upsurge, which is a well-known problem that can be eliminated by estrogen replacement therapy. Because the outbreak occurs in the central nervous system (CNS) and em_652 is difficult to reach the CNS (attached data), estrogen administration is generally expected to control the heat without being disturbed by the SERM. On the other hand, SERM eliminates the side effects of estrogen in other areas, especially breast and uterine cancer. In fact, the addition of EM-652 to estrogen inhibits the stimulating effects of estrogen on the mammary gland and uterus, whereas in other tissues, EM-652 can exert its own beneficial effects, such as on bones, where it partially recovers The effect of oophorectomy on bone mineral density. EM-652 is considered to have no adverse effects in any respect, and thus it should have a significant beneficial effect in avoiding and treating breast and uterine cancer. This data shows that the addition of EM-652 blocks the stimulation of estrogen on the mammary gland and the uterus (Examples 4, 8 and 1), while in other tissues, Εμ·652.HC1 exerts its beneficial effects. For example, in the bones (Example 5), the paper size applies to the Chinese National Standard (CNS) A4 specification (ii〇X 297 public) (please read the notes on the back and fill out this page) ήπ. -Line - Ministry of Economy Intellectual Property Office Staff Consumer Cooperative Printed 1293251 A7 B7 V. Inventive Note (23), EM-652 partially restored the effect of oophorectomy on bone mineral density. Like the already commercialized Raloxifene for the treatment of postmenopausal women with osteoporosis. In fact, Raloxifene was found to be 3 to 10 times less likely to avoid BMD loss in rats than EM-652 (Martel et al, J Steroid Biochem Molec Biol 2000: 74, pp 45-56). Although the role of SERM for BMD, such as other SERMs like Raloxifene, is not as complete as estrogen, the fractures observed in postmenopausal women have been found to be associated with estrogen and SERM. Raloxifene has the same effect. It is therefore expected that although BMD cannot be completely recovered via EM-652 or other SERMs, its effect on the fracture, which is the most important response parameter, is as important as the effect seen after the use of estrogen. In addition, it is highly likely that, as suggested, BMD measurements do not provide a complete description of the effects of compounds on bone physiology. An important aspect is that when treated with SERM, it has a beneficial effect on bones, a combination of SERM and estrogen. Primarily used to block the heat and reduce the risk of breast and uterine cancer associated with estrogen alone. The preferred SERMs discussed herein are related to: (1) all affected by the present invention. The disease; (2) the use of a disease for treating and preventing disease; and (3) preferred pharmaceutical compositions and kits. A patient who is at risk of treating or reducing the onset of a particular disease is one who has been diagnosed with or susceptible to the disease. Unless otherwise stated elsewhere, the preferred activating compound of the present invention applies the Chinese National Standard (CNS) A4 specification (210 X 297 mm) to the paper scale ------------- ------Set--------- Line (please read the notes on the back and fill out this page) 27 1293251 A7 V. Description of invention (24) Wisdom employee consumption printing dose (dosing drug dependence and Form) is the same as that used for the treatment and rib disease. The dosages for each activation element discussed herein are the same as those used for the disease being treated (or the same as those used to reduce the onset of disease). The dosage herein refers to the weight of the activating compound that is not affected by the pharmaceutical adjuvant, diluent, carrier or other material, unless otherwise stated or apparent from the context, although such added materials are included as desired. Among them, as shown in the example here. Any dosage form (capsule, tablet, injection or otherwise) conventionally used in the pharmaceutical industry is suitable for use herein and the term "auxiliary", "diluent," or "carrier, including, for example, such as they are typically included In this case, the non-activated starting material, and the activated starting material, are used in the same dosage form. For example, typical capsules, pills, enteric coatings, solid or liquid diluents or adjuvants, flavorings, preservatives or other analogs that may be included. All of the activating starting materials for the treatments discussed herein can be expressed by the formula of the pharmaceutical composition. The pharmaceutical composition also includes _ or a plurality of other activating materials. Optionally, they can be administered separately, but at the same time, so the patient can ultimately improve blood levels or enjoy the benefits of each activated ingredient (or strategy). In some preferred embodiments of the invention, for example, ' or a plurality of activated starting materials will be represented by the formula for a single pharmaceutical composition. In other embodiments of the invention, the present invention provides - a kit comprising at least two separate containers, wherein at least one of the contents of the container is the same or different from the contents of at least one of the other containers, the other The container is the activated starting material contained herein. Page Line This paper scale applies to China National Standard (CNS) A4 specifications Curry to 7 public hair -28 1293251

V. INSTRUCTIONS (25) Economy, Intellectual Property Office Staff Consumption Cooperation: The combination treatments discussed here also include the use of an activated (combined) material in the manufacture of a medicament for treating disease (or reducing disease risk). The treatment of the discussion or avoidance of the disease' includes further activation of the starting material in accordance with the combination of the invention. For example, in a specific embodiment, the present invention provides a SERM for use in the preparation of a medicament, which is combined with an estrogen and an estrogen-converted prodrug in a living body for treating any disease in which the combination therapy is believed to be effective. (ie osteoporosis, cardiovascular disease, hyperbilirubinemia, hyperlipidemia, atherosclerosis, hypertension, tamsin resistance, diabetes, obesity, craze, night sweats, irregular menstruation, Azheimer's Symptoms, cognitive problems, any signs of menopause and dryness of the sheath). In another embodiment, the invention provides for the use of an estrogen selected from the group consisting of 17 々 estradiol, 17 estradiol ester (ie, benzoic acid, cypionate, diheptane). Acid salts, valerate, etc.), estradiol, 17 α-estradiol ester, female sterol, female sterol ester 'female, female ester, conjugated estrogen, euquinoline (equilin) , eciuilin ester, 17 α -ethynyl estradiol, 17 α -ethynyl estradiol g, dilute female alcohol, mestranol, mestreol A group consisting of (mestranol esters), DES, phytoestrogens, tibolone, and acetylene glycol, used to prepare a medicament for use in combination with SERM to treat any disease in the same temple. Estrogen is widely known to stimulate breast epithelial cell proliferation, and cell proliferation itself is thought to increase the risk of cancer by accumulating random genetic errors that can cause tumor formation (Preston Martin et al., Cancer. Res. 50: 7415-21 , 1990). Based on this concept, anti-estrogen has been introduced into this paper scale for the Chinese National Standard (CNS) A4 specification (210 X 297 mm) 29 ------------- I--- --丨定------I-- (Please read the note on the back and then fill out this page) A7 1293251 __ B7_ V. Description of invention (26) to reduce the rate of estrogen-stimulated cell differentiation to avoid breast cancer The goal of the occurrence. Loss of ovarian circulation found in Sprague-Dawley mothers 10 months later is accompanied by increased serum estrogen and prolactin levels and reduced serum androgen and progesterone concentrations . (Lu et al. 5 61st Annual Meeting of the Endocrine Society 106 (abst. #134), 1979; Tang et al, Biol. Reprod. 31: 399-413, 1984; Russo et al., Monographs on Pathology of Laboratory Animals: Integument And Mammary Glands 252-266, 1989; Sortino and Wise, Endocrinology 124: 90-96, 1989; Cardy, Vet. Pathol. 28: 139-145, 1991). These hormonal changes spontaneously in aging mothers are associated with multifocal hyperplasia and increased secretory activity of glandular/cochlear tissue as well as ductal dilatation and cyst formation (Boorman et al., 433, 1990; Cardy, Vet Pathol. 28: 139-145, 1991). It is worth mentioning that proliferating and neonatal rat mammary gland changes are often accompanied by an increase in the levels of estrogen and stimulating hormone (Meltes, J. Neural. Transm. 48: 25-42, 1980). Treatment with EM-800 (a SERM of the invention) induces atrophy of the mammary gland, characterized by a decrease in the volume and number of lobular structures, and no evidence of secretion activity, demonstrating the potent antiestrogenic activity of EM-800 in the mammary gland. (Luo et al., Endocrinology 138: 4435-4444, 1997) 雌 Estrogen is currently known to lower serum cholesterol but may or may not affect serum triglyceride levels (Love et al., Ann. Intern. Med. 115: 860-864, 1991; Walsh et al., New Engl J. Med. 325: 1196-1204, 1991; Barrett-Connor, Am. J. Med. 95 (Suppl· This paper scale applies to Chinese national standards (CNS )A4 size (210 X 297 mm) (Please read the notes on the back and fill out this page)

Ministry of Economic Affairs, Intellectual Property Office, Employees' Consumption, 阼中中30 1293251 A7 B7 V. Invention Description (27 5A): 40S-43S, 1993; Russell et al., Atherosclerosis 100: 113-122, 1993; Black et al., J· Clin. Invest. 93: 63-69, 1994: Dipippo et al, Endocrinology 136: 1020-1033, 1995: Ke et al, Endocrinology 136: 2435-2441, 1995). Figure 3 shows that EM-800 has a dual role in causing hypocholesterolemia and triglyceride in rats, thus showing its unique effect on serum lipids. EM-800 is significantly different from other SERMs, such as It is Tamoxifene (Bruning et al, Br. J. Cancer 58: 497-499, 1988; Love et al, J. Natl. Cancer Inst 82: 1327-1332, 1990; Dipippo et al, Endocrinology 136: 1020-1033, 1995; Ke et al, Endocrinology 136: 2435-2441, 1995), Droloxifene (Ke et al, Endocrinology 136: 2435-2441, 1995), and Raloxifene (Black et al. , J. Clin. Invest. 93: 63-69, 1994). Therefore, it is generally believed that the combination of estrogen and EM800 should still retain the effect of low cholesterol and triglyceride caused by EM-800, so it is proposed that this combination can have a beneficial effect on serum lipids. Read the notes on the back and fill out this page. It is worth noting that the serum lipid profiles of rats and humans are significantly different. However, because estrogen receptor-mediated mechanisms are involved in estrogen and antiestrogens-induced hypocholesterolemia (Lundeen et al., Endocrinology 13 8: 1552-155 8, 1997), rats are still a useful The model is used to study the cholesterol-lowering effects of human estrogens and "antiestrogens." We also studied the new anti-estimulus paper scale by applying the combination of two drugs. The Chinese National Standard (CNS) A4 specification (210 X 297 mm) 31 A7 1293251 BT^___ V. Invention Description (28) (EM-800) and sex steroid precursor (DHEA) strongly inhibited human ZR-75-1 breast cancer xenografts in nude mice. Figures 2 and 3 show that DHEA itself, at the dose used, produces 5 肿瘤 for tumor growth. /0 to 80% inhibition, and tumor growth achieved by low dose anti-estrogen is almost completely inhibited without being affected by DHEA. As shown in Figure 4, similar effects of EM-800 and progesterone, :\1? eight pairs of E2 promoting DMBA-induced breast cancer growth were also observed in ovariectomized rats. The limits of bone mineral density (BMD) measurements are currently known. For example, BMD measurements showed that treatment with steroid anti-estrogen ICI 182780 did not cause any change (Wakeling, Breast Cancer Res. Treat. 25: 1-9, 1993), whereas inhibition was observed using histomorphometry. Change (Gallagher et al., Endocrinology 133: 2787-2791, 1993). Similar differences have also been reported with Tamoxifene (Jordan et al, Breast Cancer Res. Treat. 10: 31-35, 1987; Sibonga et al, Breast Cancer Res. Treatm. 41: 71-79) , 1996) o It must be stated that the reduction in bone mineral density is not just an anomaly associated with reduced bone strength. (Guidelines for preclinical and clinical evaluation of agents used in the prevention or treatment of postmenopausal osteoprosis, Division of Metabolism and Endocrine Drug Products, FDA, May 1994). Therefore, it is important to analyze changes in the biochemical parameters of bone metabolism caused by various compounds and treatments in order to gain a better understanding of their effects. This paper scale applies to China National Standard (CNS) A4 specification (210 X 297 mm) (please read the note on the back and fill out this page) 丨线_ Ministry of Economic Affairs Intellectual Property Bureau employees consumption cooperation Du printing system # wear 32 economy Ministry of Intellectual Property Bureau Staff Consumer Cooperative Printed 1293251 A7 B7 V. INSTRUCTIONS (29) It is important to note that the combination of DHEA and EM-800 has an unexpected beneficial effect on important bone biochemical parameters. In fact, DHEA alone does not affect the index of bone resorption: the ratio of urinary hydroxyproline/creatinine. In addition, the effect of DHEA on each mother's urine or filling and draining is not detected (Luo et al., Endocrinology 138: 443 5-4444, 1997). The EM-800 reduced the urine hydroxyproline//creatinine ratio by 48%, and similar to DHEA, did not see the effect of EM-800 on i bow or fill in each urine. Furthermore, EM-800 has no effect on serum alkaline luciferase activity, which is an indicator of bone formation, while DHEA increases this parameter by 75% (Luo et al., Endocrinology 138: 4435-4444, 1997) ° One of the unexpected effects of DHEA and EM-800 is related to the ratio of urinary hydroxyproline/creatinine, which is an indicator of bone resorption. When DHEA and EM-800 are used in combination, The ratio will decrease by 69%, which is statistically different from the 48% inhibition of EM-800 alone or the use of DHEA alone (p<0.01). Therefore, the addition of DHEA to EM-800 increased the inhibition of bone resorption by 50%. The most important % is that another unexpected effect of adding DHEA to EM-800 is a nearly 80% reduction in urine, medium calcium and phosphorus, respectively (from 2.15 to 1.51 to 0.75 ± 0.75 /zmol / 24h). /100g (ρ<〇.〇1)) and a 55% reduction (from 132.72 soil 6.08 to 59.06 士4.76//111〇1/24}1/10 (^(?<0.01)), (1^〇 Etc., Endocrinology 138: 4435-4444, 1997) The standard of the paper is applicable to the Chinese National Standard (CNS) A4 specification (210 X 297 mm) -------------Installation---- ----Book---------Line (please read the back of the note first and then fill out this page) 33 [293251 V. Inventions (30) A7 B7 Table 1 Group of urine serum calcium ( Mol/24h/100g) Phosphorus (mmol/24h/100g) HP/Cr (mmol/24h/100g) tALP (IU/L) Control group 23_17± 1·55 132.72 soil 6·08 13·04±2·19 114.25士 14.04 DHEA (lOmg) 25·87±3.54 151.41±14.57 14.02 soil 1.59 198.38±30.76* EM-800 (75mg) 17.44±4.5 102.03±25.13 6·81±0·84** 114.11±11.26 DHEA+EM-800 3·71±0.75** 59·06±4·76** 4.06 soil 0.28** 204.38 soil 14.20** (please read the notes on the back and fill out this page) Interesting and It should be noted that simultaneous treatment of DHEA does not prevent EM-800 from strongly inhibiting serum cholesterol (Luo et al., Endocrinology 138: 4435-4444, 1997) ° when Raloxifene and similar compounds avoid bone loss and reduce In serum cholesterol (like estrogen), it must be mentioned that when Raloxifene and Premarin are compared to BMD, Raloxifene has less effect on BMD than Pimalin. (Premarin) is so strong (Minutes of the Endocrinology and Metabolism Drugs. Advisory Committee, FDA Thursday, Meeting #68, November 20th 1997). Ministry of Economic Affairs, Intellectual Property Bureau, Staff Consumption Cooperative Printed It is generally believed that the bone loss observed in women during menopause is related to the rate of bone resorption, and bone resorption cannot be fully compensated by the secondary increase in bone formation. In fact, both the parameters of bone formation and bone resorption are increased in osteoporosis, and both absorption and formation are inhibited by estrogen replacement therapy. Therefore, it is generally believed that the inhibition of osteogenic formation by estrogen substitution is derived from the coupling mechanism of bone resorption and bone formation. Therefore, the reduction of bone resorption induced by primary estrogen will promote the reduction of bone formation. The paper is applicable to the Chinese National Standard (CNS). A4 size (210 X 297 public) 34 1293251 A7 B7 V. Description of invention (31) (Parfitt » Calcified Tissue International 36 Suppl. 1: S37S45, 1984) The strength of the cancellous bone and the resistance to the subsequent fractures are not just dependent The amount of cancellous bone depends on the microstructure of the small column, which is determined by the number, volume and distribution of the small columns. Loss of ovarian function in postmenopausal women is accompanied by a marked reduction in total small column bone volume (Melsen et al, Acta Pathologica & * Microbiologica Scandinavia 86: 70-81, 1978; Vakamatsou et al., Calcified organization International 37: 594- 597, 1985), mainly related to the reduction in number, and to a lesser extent to the width of the small column (Weinstein and Hutson, Bone 8: 137-142, 1987). In order to promote the combination treatment of the present invention, any For the purposes of the present discussion, the present invention provides several pharmaceutical compositions comprising a SERM and an estrogen in a single composition for administration. The composition can be applied to a variety of conventional administrations including, but not limited to, oral administration, subcutaneous injection, intramuscular injection, or transdermal administration. In another embodiment, the present invention provides a set comprising one or more SERMs and estrogens in separate or identical containers. When used for the treatment or prevention of osteoporosis, the above-mentioned drugs, compositions and kits may further comprise a bisphosphonate compound. The kit may comprise suitable materials for oral administration, such as tablets, capsules, sugar aggregates and the like, or may be used for transdermal administration, such as ointments, lotions, gels, creams, long-acting patches. Cloth and other analogues. Applicants believe that estrogen, SERM and sex steroids can be applied to developing osteoporosis, hypercholesterolemia, hyperlipidemia, atherosclerosis, hypertension, insulin resistance, diabetes, obesity, AIDS This paper size applies to the Chinese National Standard (CNS) A4 specification (210 X 297 mm) -------------Htv--- (Please read the note on the back and fill out this page)

T Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printing 35 1293251

Ministry of Economic Affairs, Intellectual Property Bureau, employee consumption cooperative, printing

Hyperthermia, as well as treating and/or reducing the incidence of hot and night sweats. The active ingredients of this month (whether hormones, SErm or precursors or others) can be formulated and administered in a number of different ways. When administered together in accordance with the present invention, the activated material can be administered simultaneously or separately. The activated material for transdermal or transmucosal administration is preferably from 0.01 to 2% by weight, and more preferably between 2% and 1% by weight, based on the weight of the pharmaceutical composition. 17 stone-estradiol, feminine, conjugated estrogen should be 〇〇1% to 1% 'DHEA or 5-diol should be at least 7% concentration for transdermal administration. Optionally, the activating material can be placed in a transdermal patch of a structure known in the art, e.g., such as the structure of European Patent No. 0279982. When formulated with an ointment, lotion, gel, cream or the like, the activating compound is admixed with a suitable carrier which is compatible with human skin and mucosa and which enhances the penetration of the compound. Through the skin or mucous membranes. Suitable carriers known in the art include, but are not limited to, Klucel HF and Giaxal base. Some carriers are commercially available, for example, Glaxal base is commercially available from Glaxal Canada Limited Company. Other suitable carriers are available from K〇Uer and Buri, s.T.PPharma 3(2), 115_124, 1987. The carrier is preferably an activating starting material which is used in a concentration which is soluble at the temperature at that time. The carrier must have sufficient viscosity so that the inhibitor can be maintained in a localized area where the composition of the skin or mucous membrane is applied. It is sufficient to allow the precursor to pass through the skin or a local area of the mucosa without flowing or evaporating for a period of time. And enter the blood stream to play the paper size applicable to the Chinese National Standard (CNS) A4 specifications (210 X 297 public) (please read the back of the note before filling this page) - · . line · -n 1 · 1 36 A7 1293251 V. INSTRUCTIONS (33 Clinical effects. The carrier is typically a mixture of several ingredients, such as a solvent that is acceptable for learning and a thickening agent. A mixture of organic and inorganic solvents contributes to hydrophilicity and Lipophilic solubility, such as water and alcohols such as alcohol. The preferred steroid precursor is dehydroepiandrosterone (DHEa) (available from

Diosynth Inc., Chicago, Illinois, USA). The carrier may also include various additives commonly used in ointments and lotions as well as in known cosmetic and pharmaceutical arts. For example, aromatic antioxidants, perfumes, gelling agents, thickening agents such as a few methylcelluloses, surfactants, stabilizers, softeners, coloring agents, and the like may be added. When used in systemic diseases, the area of application on the skin should be altered to avoid excessive local concentrations of the activated material and the skin may be over-stimulated by the activated material. The treatment according to the invention can be applied to the continuation treatment of uncertainty. Estrogen compounds, SERM compounds and/or sex steroid precursors and/or bisphosphonates may also be administered, via the oral route, and may be formulated with conventional pharmaceutical adjuvants such as dry-sprayed lactose, microcrystalline cellulose and magnesium. Stearate is incorporated into tablets or capsules for oral administration. The activating substance can be made into a tablet or made into a granulated sugar core via a solid, powder carrier material, such as sodium citrate, calcium carbonate or dicalcium phosphate, and a combination such as polyvinylpyrrolidone, gelatin or a cellulose derivative. It may also be possible to add a lubricant such as magnesium stearate, sodium lauryl sulfate, "Carb〇wax", or ♦ ethylene glycol. Of course, taste can also be added to the example of oral administration. Improve the substance. ------II ^ ----I--I · f Please read the notes on the back and fill out this page.) Printed by the Consumer Intellectual Property Office of the Ministry of Economic Affairs

1293251 A7 ------- B7 V. INSTRUCTIONS (34) In a further version, it is also possible to use a capsule, such as hard gelatin, and a lycerine containing a softening or plasticizing agent. Sealed soft gelatin capsules. The capsules contain an activating material, preferably in a particulate form, for example mixed with a filler such as lactose, sucrose, mannitol, starch, such as potato starch or amylopectin, a cellulose derivative or a highly dispersed tannic acid. In soft gelatin capsules, the activating substance is preferably dissolved or suspended in a suitable liquid, such as vegetable oil or liquid polyethylene glycol. The lotion, ointment, gel or emulsion should be completely rubbed into the skin, so it is apparent that there is no excess and the skin in this area should not be rinsed 'percutaneous penetration preferably occurs until at least 4 hours After that, it should be at least 6 hours. A transdermal patch can be used to deliver the precursor in accordance with known techniques. It usually needs to be applied for a longer period of time, for example 4 days, but the activated raw material contacts a smaller surface area to allow slow and continuous delivery of the active material. A variety of transdermal drug delivery systems that have been developed and are already in use are suitable for use in delivering the activated materials of the present invention. The rate of release is typically known by diffusion of a matrix or by activating a membrane through a control membrane to control the mechanical state of the transdermal device in rats, and is, for example, in U.S. Patent No. 5,162,037, Nos. 154,922, 5,135,480, 4,666,441, 4,624,665, 3,742,951, 3,797,444, 4,568,343, 5,064,654, 5,071,644, 5, 〇71,657. The above disclosed persons are incorporated herein by reference to the Chinese National Standard (CNS) A4 specification (210 X 297 mm) --------1---Ί --- (Please read the back first) Note: Please fill out this page) · • Line - Ministry of Economic Affairs Intellectual Property Officer X Consumer Consolidation:; mention 38 1293251 A7 V. Invention Description (35 Other Moon Hall, data from European Patent No. 279982 and British Patent No. 2185187 The device may be of any general type known in the art, including an adhesive matrix (IV) storage transdermal delivery device. The device may comprise a drug-containing matrix that incorporates fibers that absorb the activated material and/or carrier. In the device, the reservoir can be defined in a transdermal device by a polymeric film that does not penetrate the carrier and the activated material. The device itself maintains contact of the activated material with the desired topical skin surface. In the device, The carrier viscosity of the activated material is less than that of the emulsion or gel. A solvent system for a transdermal device may include, for example, oleic acid, linear alcohol lactate and dipropylene glycol or the present technology. Knowing other a solvent system. The activated material can be dissolved or suspended in a carrier. A transdermal patch for attaching to the skin can be placed in a surgical adhesive tape having a void in the middle. The adhesive is preferably lining a release. Covered to protect the adhesive prior to use. Substances that are typically suitable for release include polyethylene and coated polyethylene paper, and are preferably coated with a resin to facilitate tearing. When using the device, the release The lining can be easily removed and the adhesive can be applied to the skin of the patient. In U.S. Patent No. 5,135,480, the disclosure of which is incorporated herein by reference in its entirety, The device has a non-adhesive member that can be used to secure the device to the skin. Only SERM, estrogen, and final steroid precursors are required to be administered in one form and at a sufficient dose of each blood serum concentration. -----------Installation - (Please read the notes on the back and fill out this page) · 丨线' Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printing 39 A7 1293251 B7 _ V. Invention Description (36) Desire According to the combination therapy of the present invention, the concentration of SERM is maintained at the desired parameters, and the estrogen is maintained at the desired parameters. (Please read the notes on the back and fill out this page.) When using estradiol The serum estradiol concentration should typically be maintained between 50 and 300 ng per liter, preferably between 100 and 200 ng per liter, and optimally between 150 and 175 ng per liter. Another estrogen is also used, and the serum concentration can be changed in a known form to take into account the difference in estrogenic activity relative to estradiol in order to achieve normal estrogen levels during menopause. For mestranol, a lower concentration is required. Appropriate serum estrogen levels can also be assessed by the disappearance of menopausal symptoms. The serum concentration of the second compound (eg, EM652.HC1) of the combination therapy is typically maintained at 1 to 15/g per liter, or in some specific instances, between 2 and 10 yg per liter, or between 5 and liter per liter. 10#g. The estrogen is preferably estradiol, but may be sodium female prosulfate or any other compound that acts as an estrogen receptor co-agent. Commercially available estrogen replenishers may be used when administered separately, such as the "PREMARIN" Ministry of Economic Affairs Intellectual Property Office employee consumption partnership available from Ayerst (St-Laurent, Quebec, Canada). DHEA is a preferred sex steroid precursor, and although DHEA-S and the analogs discussed below are particularly effective, the reasons are set forth below. For a typical patient, a suitable estrogen dose to achieve a desired serum concentration by oral administration is between 0.3 and 2.5 mg per 50 kg body weight of PRJEMARIN. In the specific example of the present invention, the female The hormone can be 17 / 3 · Estradiol is administered percutaneously on a patch called "ESTRADERM", which can be purchased from CIBA, where each dose is between 50% per body weight per paper weight. National Standard (CNS) A4 Specification (210 X 297 mm) 40 1293251 A7 B7 Ministry of Economic Affairs Intellectual Property Bureau Staff Consumer Cooperative Printed V. Invention Description (37) Between 0.05 and 0.2 mg. 17-Estradiol valerate is commercially available under the trade name "DELESTOGEN", available from Squibb, which is administered by injection. Other preferred estrogen drug products of the invention are: 17/3-estradiol. Patch, available under the trade name CLIMAARA, available from Berlex Canada, or under the trade name VIVELLE, available from Novartis Pharma; sheath device containing 17/5-estradiol, trade name £8丁十一川0, available from Pharmacia &Upjohn; gel containing 17 /5 -estradiol, trade name ESTROGEL, available from Schering; emulsion containing 17 /3 -estradiol under the trade name ORTHO DINESTROL Available from JANSSEN-ORTHO. In some embodiments, the estrogen is preferably administered orally. For example, the commercially available product of micronized 17 /5 -estradiol is ESTRACE, available from Roberts; the trade name of ethinyl estradiol is ESTINYL, available from Schering Canada; the trade name for female sulphate is OGEN. Available from PHARMACIA UPJOHN. In some embodiments, a mixed estrogen/androgen compound is preferred over estrogen. One of the compounds is tibolone [7α,(7α)·17-hydroxy-7-methyl-19-pregnant-5(10)-ene-20-yn-3-one; US Patent No. 3,340,279 (1967); U.S. Patent No. 3,475,465 (1969) and J. de Visser et al., Arzneimittel-Forsch, 34, 1010, 1984, Introduction to Endocrinology, available under the trade name LI VIAL, available from ORGANON (The Netherlands) ) purchased. Pharmaceutical products containing a mixture of estrogen and progesterone or androgen. This paper scale applies to the Chinese National Standard (CNS) A4 specification (210 X 297 mm). 41 --------------- ( Please read the notes on the back and fill out this page. 言 r 幺v> A7 1293251 B7 _ V. Invention description (38) is also preferred. The drug is marketed under the trade name ESTRACOM and is available from Novartis Pharma or under the trade name CLIMACTERON, available from Sabex. The transdermal or transmucosal delivery system of the present invention can also be used as a new and improved delivery system to prevent and/or treat osteoporosis or other diseases. Any manufacturer's recommendations for estrogen to achieve the desired effect can be used. Suitable dosages are known in the art. Any compound or mixture of compounds having estrogenic or similar or co-active activity on an estrogen receptor or the like can be used in accordance with the present invention. (eg phytoestrogens, synthetic estrogens, etc.) A selective estrogen receptor modulator of the invention is a molecular formula having the following characteristics: a) 2 aromatic rings separated by 1 to 2 carbon atoms , 2 rings may be substituted by a hydroxyl group or a substituent converted into a hydroxyl group in vivo, or may not be substituted; and b) 1 side chain having an aromatic ring and a tertiary amine function or a salt thereof. A preferred SERM of the present invention is the EM-800 reported in PCT/CA96/00097 (W0 96/26201). The molecular structure of the EM-800 is (please read the notes on the back and fill out this page) 丨线 - Printed by the Ministry of Economic Affairs, Intellectual Property Bureau, Staff Consumer Cooperative

This paper scale applies to the Chinese National Standard (CNS) A4 specification (210 X 297 mm) 42 When the company is smart, it is the employee's consumption order;;! :一印纠农 1293251 A7 __B7 V. Description of the invention (39) Another preferred SERM system of the present invention is EM-01538 ··

ΕΜ·1538 (also known as EM-652.HC1) is a hydrogenate for the potent anti-estrogen EM-652. Compared to EM-800, EM-1538 is a simpler and easier to synthesize salt. It is generally believed that it will produce the same activating compound in vivo. Other preferred SERMs of the present invention include it, moxifen (Ding &! 11 (^[61^)((2)-2-[4_(1,2-diphenyl 1-butenyl))) Base]-:^,:^-dimethylethaneamine) (available from Zeneca, UK), toremifene ((Z)-2-[4-(4-gas-1,2- Diphenyl-1-butenyl)phenoxy> N,N dimethylthyristine) (available from Orion-Farmos Pharmaceuticla, Finland or Schering-Plough), Droloxifene (E )-3-[l-[4-[2-(dimethylamine)ethoxy]phenyl]-2]phenyl-1-butenyl]phenol) and €?-336,156 (Lasofosie) Fen (1^3〇€〇\丨€61^)) (cis-111-[4'-°bit each bite ethoxyphenyl]-2S-phenyl-6-radio-1,2,3 , 4-tetrahydronaphthalene D-(-)-tartrate) (Pfizer Inc., USA), Ral〇xifene ([2-(4-phenylphenyl)·6-hydroxybenzo[b] Thiet-3-yl][4-[2-(l-hexahydro.bipyridyl)ethoxy]]]-methanone hydrochloride (En Lilly and c〇·, USA), LY 33 5563 (6-hydroxy_3_[4-[2-(l-hexahydroindolepyridinyl)ethoxy]phenoxy]-2-(4-hydroxyphenyl)benzo[b] spur hydrochloric acid) and LY 353381 (Arzoxifen) Fene),6-Applicable to China National Standard (CNS) A4 specification (210 X 297 mm) by this paper scale 43 -------------- Strong clothes -------- --------- Line f Jing read the phonetic transcription on the back? Matters and fill out this page} A7 1293251 B7_ V. Invention description (4G) (Please read the note on the back and fill in this page) -[4[2-(l-hexahydronyl)ethoxy]phenoxy]-2-(4-methoxyphenyl) benzo[b]thiophene hydrochloride) (Eli Lilly and Co., USA), Idoxifene ((E)-l-[2-[4[l-(4-indolyl)-2-indolyl-1·butyl)-yloxy]ethyl吼 吼 ) () (SmithKline Beecham, USA), Levormelomifene (3,4-trans-2,2-dimethyl-3-phenyl-4-[4-(2-( 2(Pyrrolidin-1-yl)ethoxy)phenyl)-7-methoxychroman] (Novo Nordisk, A/S, Denmark) This compound is disclosed by Shalmi et al. in WO 97/25034 , WO 97/25035, WO 97/25037, WO 97/25038; and Korsgaard et al, WO 97/25036), GW 5638 (as described by Willson et al., Endocrinology, 138(9), 3901-3911, • Line · Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printing 1 997) and anthracene derivatives (Miller et al., European Patent No. 0802183A1) and TSE 424 developed by Wyeth Ayers (USA) and (American home products corporation) disclosed in Japanese Patent No. 10036347 and Non-steroidal estrogen derivatives as described in WO 97/32837. Also included is iproxifene (TAT 59; (E)-4-[l-[4-[2-(dimethylamine)ethoxy)phenyl] from Taiho (Japan) -2-[4·(1-methylethyl)phenyl]-1-butenyl]phenol dihydrogen phosphate), FC 1271 from Orion (Finland) ((Ζ)-2-[4-( 4-A gas-1,2-diphenyl-1-butenyl)phenoxy]ethanol), HMR 3339 and HMR 3656 from Hoechst Marion Roussel, SH 646 from Schering AG, Germany, from Wyeth Ayerst ( USA) ERA 923, LY 335124 and LY 326315 from Eli Lilly (USA). Any manufacturer's recommendations that the SERM uses to achieve the desired effect can be used. Suitable dosages are known in the art. Any commercially available paper size is subject to the Chinese National Standard (CNS) A4 specification (210 fathers 297 mm). 44 Ministry of Economic Affairs Intellectual Property Officer X. Consumers Co., Ltd. Printed 1293251 A7 _____B7___ V. Description of invention ( 41) Other non-steroidal antiestrogens can be used in accordance with the present invention. Any compound having similar SERM activity (e.g., Raloxifene) can also be used. According to the SERM administration of the present invention, when administered orally, a dose ranging from 0.01 to 10 mg/kg body weight per amp (preferably Λ 0·05 to 1.0 mg/kg) is preferably administered, per 曰5 mg is administered, in particular 10 mg per sputum, ^ The dose is equally divided into 2 points according to the average body weight of one person. When administered parenterally (i.e., intramuscularly, subcutaneously or transdermally), a dose ranging from 0.003 to 3.0 mg/kg body weight per amp (preferably 0.015 to 0.3) is preferably administered. 11^/1111), 1.5111 § per sputum, special case 3111 § 'The average weight of one person is preferably divided into two equal doses. Preferably, the SERM is administered with a pharmaceutically acceptable diluent or carrier as described below. In the combination therapy of the present invention, preferred bisphosphonates for use in the treatment of osteoporosis as an activating starting material include Alendronate [(4 amine small hydroxybutylidene) bisphosphonate, Disodium salt, hydrate]' trade name Fosamax, available from Merck Shape and Dohme; Etidronate [(l-hydroxyethylidene) diphosphonic acid, 2,2,-, amine Diethanol]'s trade name Didrocal and Didronel, available from Procter and Gamble; ci〇dronate [(diqiethylene)diphosphonic acid, disodium salt], trade name Bonefos, available from Rhone-Poulenc Rorer is available under the trade name Ostac, available from Boehringer Mannheim; and Pamidronate (3-amino-1hydroxypropyl)diphosphonic acid, disodium salt) under the trade name Aredia Available from Geigy; Resino's paper size applies to China National Standard (CNS) A4 specification (210 X 297 mm) -------------^------ --^---------^ (Please read the notes on the back and fill out this page) 45 1293251

V. INSTRUCTIONS (42) When the Ministry of Economic Affairs is wise, the smasher 11 Shaw 6WQ乍: iLfNii (Risedr〇nate) (l-hydroxy-2-(3_acridinyl)ethylidene diphosphonic acid monosodium salt) , is currently in clinical development. Any other commercially available bisphosphonate can be used in accordance with the present invention, all dosages being as recommended by the original manufacturer. Similarly, sex steroid precursors can be administered in dosages as suggested by the prior art, preferably using a dose that allows a healthy male or a premenopausal adult female of 20 to 30 years to return to a circulating level. With regard to all doses recommended herein, the attending clinician should monitor the response of individual patients and adjust the dose accordingly. EXAMPLES Example 1 In the mammary gland, male hormones are produced from the precursor steroid dehydroepiandrosterone (DHEA). Clinical evidence indicates that androgen has a inhibitory effect on breast cancer. On the other hand, estrogen stimulates the development and growth of breast cancer. We studied the effect of DHEA alone or in combination with the newly stated pure antiestrogens EM-800 in ovariectomized nude mice on xenograft tumor growth in human breast cancer cell line zr-75-I. The mice received a subcutaneous injection of 〇5/g g female hormone (an estrogen hormone) daily after ovariectomy. EM-800 (1 5, 50 or 100 // g) is given orally daily. DHEA (total dose 〇·3, ι·〇 or 3.0 mg) was applied to the dorsal epidermis every 2 times. It was used alone or in combination with an oral dose of EM-800 per 15 #g. The tumor volume corresponds to the post-treatment change and is periodically evaluated relative to the measurement made on day 1. At the end of the experiment, the tumors were excised and weighed. The mice that received the female hormone alone were compared with the mice that did not receive the female pigment. This paper scale applies the Chinese National Standard (CNS) A4 specification (210 X 297 mm). Read the notes on the back and fill out this page) - · Line · 46 A7 1293251 ___B7___ V. Inventive Note (43), the tumor volume was observed to increase by 9.4 times within 9.5 months. EM-800 administration at 15, 50 or 100 // g resulted in 88%, 93%, and 94% tumor volume inhibition in female ovariectomized mice. On the other hand, doses of 0.3, 1.0 or 3.0 mg of DHEA inhibited the final tumor weight by 67%, 82% and 85%, respectively. A comparison of tumor volume inhibition can be obtained from daily use of 15. Λ // g oral dose of EM-800 with or without different transdermal doses of -DHEA. DHEA and EM-800 independently inhibited the growth of female-stimulated ZR75-1 mouse xenograft tumors in nude mice. Administration of DHEA at a defined dose does not alter the inhibition of EM_800. Materials and Methods ZR-75-1 Cell ZR-75-1 cell line was obtained from the American Type Culture Collection (Rockville, MD) and was conventionally added with 2 mM L-glutamine, 1 mM sodium pyrogluconate. The monolayer was cultured in PRMI 1640 medium with salt, 100 IU penicillin/ml, 100 mg streptomycin/ml and 10% fetal bovine serum, and cultured at 37 ° C, 95% air and 5% carbon dioxide as described. In humid air (Poulin and Labrie, Cancer Res. 46: 4933-4937, 1986, Poulin et al, Breast Cancer Res. Treat. 12: 213-225, 1988). Cells were seeded weekly after treatment with 0.05% trypsin: 0.02% EDTA (W/V). The cultured cell line used for the experiments in this report was derived from the 93rd generation of ZR-75-1 cell line. Animal female homozygous Harlansbach-Dore (nu/nu) Ehrlich mice The paper size applies to the Chinese National Standard (CNS) A4 specification (210 X 297 mm) ---------- ---装·-------Book---------Line (please read the note on the back and fill out this page) Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printing 47 1293251 A7 B7 V. INSTRUCTIONS (44) Qi Qi% ί % \ soil (athamic mice) (28 to 42 large) was obtained from HSD (Indianapolis, Indiana, USA). The mice were housed in a cage of vinyl material with a filtered air hood and kept under conditions to control the pathogen. Cages, grasshoppers and feeds are sterilized prior to use. The drinking water is sterilized and acidified to pH 2.8 without restriction. The cells were inoculated before the tumor cells were inoculated, and the ovaries on both sides of the mice were simultaneously excised (ovx). The tumor cells were inoculated intraperitoneally into each animal with 25 ml of anesthetic (pentanol··0.8 g/100 ml 0.9% NaCl; and three Performed in anesthesia with ethanol: 2g/100 ml 0.9% NAC1). After monolayer cells were treated with 0.05% transcriptase/0.02% EDTA (w/v), zRjsq cells with a cell density of 1·5χ106 in logarithmic growth phase were collected. The cells were suspended in 1 ml of a culture medium containing 25% Martigel, and the cells were inoculated subcutaneously on the two sides of the animal using a 1 inch long 20 gauge needle as previously described (Dauvois et al., Cancer) Res. 51: 3131-3135, 1991). In order to accelerate tumor growth, each animal received subcutaneous injection of g# g of estradiol (E2) for 5 weeks. The estradiol was contained in 0.9% Nacl, 5% ethanol, and 1% gelatin. In the agent. After the palpable ZR-75-1 tumor appeared, the diameter of the tumor was measured with a caliper, and mice having a tumor diameter of 0.2 and 〇·7 cm were selected for the study. Hormone King f All animals except the OVX control group received a daily subcutaneous injection of 0 5 # g of female pigment (El) contained in 0.2 ml, 0.9% NaC 5% ethanol, and 1% gelatin. In the designated group, DHEA applies the Chinese National Standard (CNS) A4 specification (210 297 mm) for each animal 0.3, the paper scale (please read the back of the note first and then fill out this page) Line 48 A7 1293251 ______B7__ V. Description of the invention (45) A dose of 1.0 or 3.0 mg was administered twice a day in a transdermal dose of 0.02 ml to the dorsal epidermal region outside the tumor growth zone. DHEA is dissolved in 50% ethanol and 50% propylene glycol. EM-800 ((+)-7-Secondary-pentyloxy-3-(4'-dihydroxyphenyl)-4-methyl-2(4"-(2"'-hexahydroacridine Ethoxy) stupid)-2H-stupid is synthesized by the method described previously (Gauthier et al, J, Med. S Chem. 40: 2117-2122, 1997) in the intramolecular division of the CHUL Research Center. The Department of Medicinal Chemistry of the Laboratory of Molecular Endocrinology of the CHUL Research Center. EM-800 was dissolved in 4% (v/v) ethanol, 4% (v/v) polyethylene glycol (PEG) 600, l°/〇 gelatin, 0.9% (w/v) NaCl. Animals of the indicated group received daily oral doses of 15 #g, 50//g or 100" g of EM-800, alone or in combination with DHEA, whereas animals in the OVX group received vehicle alone (0.2 ml 4 % ethanol, 4% PEG 600, 1% gelatin, 0.9% NAC1). The tumor was measured once a week with a vernier caliper. Two vertical diameters (L and W) in cms were recorded, and the tumor area (cm2) was calculated using the formula: L/2xW/2x 7Γ (Dauvois et al., Cancer Res. 51: 3131-3135, 1991). The area measured after the first day of treatment was considered to be 100%, and the change in tumor volume was expressed as a percentage of the initial 'tumor area. Generally, if there is a subcutaneous tumor, it is impossible to correctly evaluate the three-dimensional volume of the tumor, and therefore, only the tumor area will be measured. After 291 days (or 9.5 months) of treatment, the animals were killed. The range of reactions is as described (Dauvois et al, Breast Cancer Res. Treat. 14: 299-306, 1989; Dauvois et al, Eur. J. Cancer Clin. Oncol. 25: 891-897, 1989; Labrie et al. , Breast Cancer Res· Treat. 33: 237-244, 1995) for evaluation. In short, some of the paper size is applicable to the Chinese National Standard (CNS) A4 specification (210 X 297 mm) -------------Installation ------- -------- Line (please read the note on the back and then fill out this page) Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printing 49 1293251 A7 ___ B7 V. Invention description (46) corresponds to those degradation greater than Or a tumor equal to 50% of their original volume; a stable response refers to a primitive volume with tumor degradation less than 50% or evolution less than 50%, and evolution refers to evolution of more than 50% when compared to the original volume of the tumor. At the end of the experiment, all animals were killed by decapitation. The tumor, uterus, and sheath were immediately removed, connective and adipose tissue removed, and weighed. Statistical analysis The statistical significance of treatment on tumor volume was assessed using ANOVA, assessed by DHEA, EM-800 and time, and repeated measurements at the beginning and end of treatment (individuals with group factors) Impact. The random blocks constituting the animals were repeatedly measured at 0 o'clock and after 9.5 months of treatment. Time is therefore analyzed as a function within the block, and two processes are treated as inter-block interactions. All interactions between the main roles are included in this model. Whether the treatment factors are meaningful and their interactions are analyzed using individuals within the group as error conditions. The data is converted to logarithms. The ANOVA-based hypothesis assumes the remainder of the variation and the normality of the homogeneity. The inductive pairing comparison was compared in the smallest sense by Fisher's test. The primary effects and interactions on body weight and tissue weight were analyzed using standard two-way ANOVA with interaction. All ANOVAs were performed using SAS software (SAS Institute, Cary, NC, USA). The significance of the difference is declared using a two-tailed test with a full level of 5〇/〇. The same range of data is used in the Kruskall-Wallis test for the same range of reaction variables (complete response, partial reaction, stable paper scale applicable to China National Standard (CNS) A4 specification (210 X 297 public) (Please read the back of the note first and then fill out this page) - Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printed 50 A7 1293251 _____El____ V, invention description (47) fixed reaction and tumor degradation) for analysis. An overall assessment of the treatment effect is completed, and the subset of results presented in Table 4 is adjusted by adjusting the critical p-value for multiple comparisons. The exact p-value was calculated using StatXaxt software (Cytel, Cambridge, MA, USA). Data are expressed as the standard deviation (SEM) of the mean of 12 to 15 mice per group of mean soil. The results are shown in Fig. 2A. In the subject of this study, human ZR-75-1 tumors were 291 days (9.5 months) after subcutaneous administration of a 0.5/zg dose of female ovarian resection in nude mice. The increase was 9.4 times, while the tumor volume of the control OVX mice receiving only the vehicle was reduced to 36.9% of the original value. Treatment of increasing doses of DHE A percutaneously resulted in progressive inhibition of E-stimulated ZR-75-1 tumor growth. Treatment of 0. 3 mg, 1.0 mg, and 3.0 mg DHEA per day for 9.5 months resulted in 50.4%, 76.8%, and 80.0% inhibition, respectively (Figure 2A). Consistent with the total tumor load, treatment with DHEA resulted in a significant reduction in the mean tumor weight remaining after the end of the experiment. In fact, the mean tumor weight decreased from 1.12±0·26 g in the control group E to the ovariectomized nude mice to 0.37±0.12 in the group of animals receiving DHEA in the daily doses of 〇·3, 1.0 and 3.0 mg, respectively. g (P = 0.05), 0.20 ± 0.06 (Ρ = · 〇〇 1) and 〇 · ΐ 7 ± 0 · 06 g (Ρ = · 0009) (Fig. 2B). When compared with the tumor volume of the control animals at 9.5 months, anti-estrogen ΕΜ-800 inhibited estrogen-stimulated tumor volume at 15"g, 50//g and 100#g doses per sputum 87.5% (P < .0001), 93.5% (P < .0001) and 94_0% (p < .0003) (Fig. 3A). The three EM-800 doses are applicable to the Chinese National Standard (CNS) A4 specification (210 X 297 mm) -------------Installation--------Set-- ------- Line f Please read the notes on the back and fill in this page.) 51 1293251 Α7 _____ Β7 V. Invention description (48) (Please read the notes on the back and fill out this page) There is no meaningful difference between the volume reductions and each other. As shown in Figure 2B, the tumor volume at the end of the 9.5 month study was reduced from 1.12 ± 0.26 g in the ovariectomized mice of the control group to 15 // g, 50 // g per day, respectively. 〇〇8 ± 0.03 g, 〇.〇3±0.01g and 0·04±0·03 g in 100 #g dose EM-800 animals (replenishment of OVX at all doses of eM-8〇〇) That is, P < 0001.) As described above, a daily oral dose of 15/g anti-estrogen EM-800 caused an 87.5% inhibition of estrogen-stimulated tumor growth after 9.5 months. The addition of three different DHEA doses did not have a significant effect on the apparent tumor volume inhibition caused by the daily 15 #g dose of DHEA (Fig. 5B). Therefore, the mean tumor weight was significantly reduced from 1.12±0·26 g in the control group of female-fungal-supplemented mice to 15//g anti-estrogen or 0.3, 1.0, and 3. 0.08 ± 0.03 g (Ρ<·〇〇〇ΐ), 〇·ιι 〇·〇4 g (ρ=·〇〇〇2), 〇·ΐ3±〇·〇7 g (A) of the DHEA combination treated animals Ρ=·0004) and 0·08±0.05 g (Ρ<·0001) (meaningful differences are not indicated in the 4 groups) (Fig. 2). The Ministry of Economic Affairs' Intellectual Property Office employee consumption cooperative printing is also interested in testing the range of reactions that can be achieved by the above specified treatment. Therefore, treatment with increasing doses of DHEA reduced (although not yet reached a statistically significant level (ρ=·〇88)) the number of tumors in progress, from 87.5% of the control group OVX animals supplemented by females to treatment 50.0%, 53.3% and 66.7% of the sputum 罝 0.3, 1 · 〇 or 3.0 mg DHEA animals (Table 4). On the other hand, the complete response increased from 女性% of female tonifying mice to 28.6%, 26.7%, and 20.0% of animals receiving DHEA at a daily dose of 0.3, 1.0, and 3.0 mg. On the other hand, the measured stable response is on the paper scale applicable to the Chinese National Standard (CNS) A4 specification (210 X 297 mm) 52 1293251

Printed by the Ministry of Economic Affairs, Intellectual Property Office, and the Consumer Cooperatives. The control Ei-fed mice and the three groups of animals receiving the above-mentioned designated agent iDHEA were 12.5%, 21.4%, 20.0%, and 13.3%, respectively. In the control group, the nest-removed mice, the ratios of complete, partial, and stable responses were 68.8%, 6.2 〇/〇, and 18.8%, respectively, and the degradation was only seen in 62% of the tumors (Table 2). ). Among the animals receiving 15 #g anti-estrogen ΕΜ-800 (Ρ=·0006) alone or in combination with 0.3 mg, 1.0 mg or 3.0 mg of DHEΑ, 29.4%, 33.3%, 26.7% and 35.3% of the tumors were respectively A complete response or tumor disappearance is achieved (Table 4). On the other hand, tumor degeneration of 35.3 〇/〇, 44. 4%, 53.3%, and 17.6% was observed in the same group of animals, respectively. There was no significant difference between the treatments of εμ_8〇〇, either alone or in combination with DHEA. There was no significant difference in the effect of DHEA or 800800 treatment on tumor weight-regulated body weight. Treatment of 〇νχ mice with feminin increased the uterine weight from 28±5 11^ in sputum control mice to 312±8 111 § (?<.01), while increasing doses of DHEA resulted in an evolutionary but relative Smaller female stimulatory inhibition, which can reach 26% (Ρ=·0008) when using the highest DHEA dose. It can also be seen in the same figure that the weight of uterus stimulated by females was reduced from 132±8 mg in mice supplemented with female mil5//g, 50//g4i〇〇#gEM-80 (^49). ±3mg, 36±2mg, and 32±1 mg (all doses vs. control, Pc.0001). 15 // g EM-800 in combination with 0.3 mg, 1·〇mg, or 3.0 mg of DHEA per oral dose 'Measured uterine weights were 46 ± 3 mg, 59 ± 5 mg and 69 soil 3 mg. This paper scales the standard (CNS) A4 specifications (210 X 297 metric) --------- ----装--------Book. (Please read the note on the back and then fill out this page) 53 A7 1293251 _________B7__ V. Description of invention (50) On the other hand 'female treatment to make the sheath weight Increased from 14 ± 2 mg in OVX animals to 31 ± 2 mg (Ρ <·〇1), and there was no meaningful effect on the addition of DHEA. Treatment of 15#g, 5〇//g or 100# g per dose After EM-800, 'box membrane heavy lotus: decreased to 23 ± 1 mg, 15 ± 1 mg, and 11 ± 1 mg, respectively (all doses to the control group, all P and pair, P < 〇〇〇 1). · 3 mg, 1·〇mg or 3.0 mg dose of DHEA and the aforementioned dose of EM-800 combination At the time of 'measurement, the sheath weights were 22 soil 1 mg, 25 ± 2 mg, and 23 soil 1 mg, respectively (all groups had no significant difference (NS·) compared to 15//g em-800). Explain that at the highest dose, ie 1曰 per 〇〇, EM-800 reduced the uterus weight reduction of female OVX animals to 'the value of the OVX control animals', and the sheath weight was different. Reduced to a lower value than the OVX control group (Ρ <·05). DHEA partially offsets the effect of EM-800 on uterine and sheath weight, probably due to its male effect. Read the notes on the back and fill out this page. ) 0 i Line - Ministry of Economic Affairs Intellectual Property Bureau Staff Consumer Cooperative Printed This paper scale applies to China National Standard (CNS) A4 specification (210 X 297 mm) 54 1293251 A7 B7 Five , invention description (51) Table 2. In nude mice, DHEA transdermal administration or EM-800 alone orally or a combination of the two used 9.5 months of response to human ZR-75-1 breast cancer xenograft (complete, partial , stability, evolution) The total number of animals. The reaction range group is completely stable. Number of evolutions_two fn(%) OVX 16 11 (68.8) 1(6.2) 3(18.8) 1(6.2) OVX+E丨(0.5" g) 16 0 0(0) 2(12.5) 14(87.5) (〇) OVX+EKO.5// g) 0.3mg 14 4 (28.6) 〇(〇) 3(21.4) 7(50.0) +DHEA 1 .Omg 15 4 (26.7) 0(0) 3(20.0) 8 (53.3) 3 .Omg 15 3 (20.0) 0(0) 2(13.3) 10(66.7) OVX+E丨(0.5# g) 15 ^ g 17 5 (29.4) 1(5.9) 5(29.4) 6( 35.3) + EM-800 S〇μ g 16 4 (25.0) 3(18.8) 5(31.2) 4(25.0) 100 V g 16 8 (50.0) 〇(〇) 3(18.8) 5(31.2) OVX + E丨(0.5 g) 0.3mg 18 6 (33.3) 0(0) 4(22.2) 8(44.4) + EM-800+DHEA 1 .Omg 15 4 (26.7) 0(0) 3(20.0) 8(53.3) 3.Omg 17 6 (35.3) 0(0) 8(47.1) 3(17.6) ----------------- (Please read the notes on the back and fill out this page) -Line - 匕二女性素; DHEA=Dehydroepianrosine; 〇VX=Ovariectomy Example 2 Ministry of Economic Affairs Intellectual Property Bureau Employees Consumption Cooperative Printed androstene-3 yS, 173-diol (5-diol ) has intrinsic estrogenic activity. In addition to this, as a precursor steroid, it can be converted into activated androgens and/or other estrogens in the peripheral internal secretory tissues. To assess the relative importance of estrogen and 5-diol estrogen components on bone, 21-week-old rat ovaries were excised and treated once daily for 2, 5 or 12·5 mg of 5- Glycol, treated alone or with anti-androgenic Flutamide (FLU, 10 mg, sc·, once per dose) and/or anti-estrogen paper size applicable to Chinese National Standard (CNS) A4 specification (210 X 297 mm) 55 Ministry of Economic Affairs Intellectual Property Office Staff Consumer Cooperative Printed 1293251 a: ___Β7 V. Description of invention (52 EM-800 (100 // g, sc, once daily) for 12 months. Bone mineral density ( BMD) was measured after 11 months of treatment. Ovariectomy (〇νχ) resulted in a 12.8% reduction in femoral BMD (ρ<〇·〇1), while in the 11 months after 〇VX, the highest dose was processed 5 -diol recovered 34.3% of BMD loss (ρ<0·01). At the same time, FLU completely prevented the stimulating effect of 5-diol on BMD of the femur' while adding EM-800 resulted in one phase compared to 5- 28.4% additional stimulation of diol alone. Simultaneous administration of 5-diol, FLU and EM-800 showed only EM-800 (27%) because of 5-diol The effect was completely blocked by FLU. Similar results were obtained from BMD of the lumbar spine, although the lumbar spine BMD of OVX rats received 12.5 mg of 5-diol, 12.5 mg of 5-diol + EM-800 alone or 5-diol+FLU+EM-800 returned to a value that was not significantly different from intact animals. Histomorphometric analysis showed that the stimulating effect of 5·diol on bone volume, number of columns and the inhibitory effect were close. The inhibition of the secondary cancellous column of the end-diameter bone end zone was abolished by FLU, but was further enhanced by EM-800. Obvious strain of serum alkaline phosphatase activity was obtained after treatment of 5-diol. The FLU that was administered at the same time recovered 57% (ρ < 0·01 vs. 12.5 mg of 5_diol alone). There was no statistically significant inhibition of the ratio of calcium to creatinine in the urine treated with diol. The highest dose of 5-diol resulted in a 23% (ρ<〇·〇1) meaningful decrease in serum cholesterol, while the addition of ΕΜ-800 reduced cholesterol by 62% (ρ<〇·〇ΐ). Shows the stimulating effect of 5-diol on bone formation and suggests that although 5-diol It is a weak estrogen, and its stimulating effect on bone formation is mainly through the action of a male hormone. Even worse, the additional stimulation of bone by ΕΜ800 and 5-diol reveals resistance (please read the back) Precautions and then fill out this page) Book · Line ' 56 1293251 Α 7 Β 7 5, invention description (53 estrogen ΕΜ-800 bone loss in rats. The cholesterol lowering activity of 5-diol and cesium-800 can be used as an interesting application to avoid cardiovascular disease. Example 3 Synthesis of a preferred compound of the present invention Synthesis of (S)-(+)-7-hydroxy-3-(4' -hydroxyphenyl)-4-methyl-2-(4"·(2", hexahydroacridine ethoxy)phenyl)-2Η-1-nonanhydroquinone hydrazine_01538 (EM-652, HC1) (Please read the notes on the back and fill out this page)

4 Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printed

ΕΜ·01538 Step A ·· BF3Et20, toluene; 100 °C; 1 hour. Step C: 3,4-Dihydro 17-pyran, p-toluene acid monohydrate, ethyl acetate The paper scale applies to China National Standard (CNS) A4 specification (210 X 297 mm) 57 A7 1293251 ___B7_ V. BRIEF DESCRIPTION OF THE INVENTION (54) (Please read the notes on the back and fill out this page) Salt; crystallize at 25 ° C under nitrogen for 16 hours and then in isopropanol. Steps D, E and F: (1) Hexahydroquinone ratio bite, toluene, Dean-Stark equipment, reflux under nitrogen; (2) 1,8 diazabicyclo[5, 4, 0] eleven -7-ene, DMF, reflux for 3 hours; (3) CH3MgCM, THF, -20 to 0 ° C and then room temperature for 24 hours; Step G, H: (lS)-(+)-10-camphorsulfonic acid , acetone, water, toluene, room temperature, 48 hours step HH: 95 ° /. Ethanol, 70 ° C, then set at room temperature 3 曰. Step HHR: recovery of the mother liquor and the step HH lotion (S)-10-camphorsulfonic acid, reflux; 36 hours, then room temperature for 16 hours. Step I: (1) DMF solution, Na2CO3, ethyl acetate; (2) Ethanol, diluted HC1; (3) Water. . Line. Printed by the Ministry of Economic Affairs, Intellectual Property Bureau, Staff Consumer Cooperative, Synthetic 2 - 4 吼 基 乳 -4- 4-ionyl-2'-(4-tetrazolium oxalyloxy) acetophenone (4). A suspension of 2,4-dihydroxy-2'-(4"hydroxyphenyl)acetamidine 3 (97·6 g, 0.4 mole) (available from Chemsyn Science Laboratories, Lenexa, Kansas) at 3,4-di Treatment with p-toluenesulfonic acid monohydrate (0.03 g, 0.158 mmole) in hydroquinone (2 1 8 ml, 3.39 mole) and ethyl acetate (520 ml) was carried out at about 25 ° C. The mixture was stirred without additional heating and under nitrogen for about 16 hours. The mixture was then washed with an aqueous solution of sodium dicarbonate (1 g) and sodium carbonate (5 g) dissolved in water (100 ml). Wash with brine (20 ml) and re-extract with 50 ml of ethyl acetate each time. Mix all organic phases and filter through sodium sulfate. This paper scale applies to China National Standard (CNS) A4 specification (210 X 297厘) 58 Ministry of Economic Affairs Intellectual Property Bureau Staff Consumer Cooperative Printed 1293251 A7 -_ __B7 V. Description of Invention (55) Solvent (about 600 ml) was removed by distillation under atmospheric pressure and isopropanol (250 ml) was added. Additional solvent (approx. 300 ml) is distilled at atmospheric pressure and isopropanol (250 ml) is added. Additional dissolution (about 275 ml) was distilled at atmospheric pressure and isopropanol (250 ml) was added. The solution was stirred and cooled at about 25 ° C, and after about 12 hours, the crystalline solid was filtered and washed with isopropanol. Dry (116.5 g, 70%). Synthesis of 4-hydroxy-4-methyl·2-(4,-[2,-hexahydroacridine]-ethoxy) stupyl 3-(4,,, four Hydrogen π-pyranyloxy) stupyl 1 tetrahydrofurfuryloxy-chroman (chr〇mane) (10). 2-tetrahydrofurfuryloxy-4-hydroxy-2,-(4, tetrahydroanthracene喃 氧 ) ) ) 醯 ) ) 4 (1 kg, 2.42 mole), 4_[2-(1 hexahydroacridine) ethoxy] acetal aldehyde 5 (594 g, 2.55 mole) (from Chemsyn Science Laboratories, Lenexa) ,

a solution obtained from Kansas) and hexahydroacridine (82 4 g, 0.97 mole) in toluene (8 L) (from Aldrich Chemical Company Inc., Milwaukee,

Wis. was obtained by refluxing with Dean-Stark equipment in nitrogen until 1 equivalent of water (44 mL) was collected. Toluene (6.5 L) was removed from the solution by distillation at atmospheric pressure. Dimethylformaldehyde (6.5 L) and 1,8-diazabicyclo[5,4,fluorene]undec-7-ene (110.5 g, 0.726 mole) were added. The solution was shaken at room temperature for about 8 hours to isomerize chalcone 8 to clormannone 9 and then to water and ice (8 L) and toluene (4 l) In the mixture. The phases were separated and the layer was washed with water (5 L). The mixed aqueous lotion was drawn with toluene (3 X 4 L). The mixed toluene extract was finally washed with brine X 4 L), concentrated to 5.5 L at atmospheric pressure, and then cooled to _ i 〇 °c. Under continuous external freezing and stirring in nitrogen, add 3?4--------------------- order--------I (please first Read the urgent notes on the back and fill out this page. 59 I293251 A7 ------B7 ... _____ V. Description of the invention (56) (Please read the note on the back and fill in the page again) A solution of magnesium THF (2.5 L, 7.5 mole) (available from Aldrich Chemical Company Inc., Milwaukee, Wis.) was maintained at a temperature below 〇 °c. After all the Grignard reagents were added, the external cooling was removed and the mixture allowed to warm to room temperature. The mixture was stirred at this temperature for about 24 hours. The mixture was again cooled to about -20 ° C and external cooling and mixing continued, and a saturated ammonium carbonate solution (200 ml) was slowly added to maintain the temperature below 20 °C. The mixture was stirred for 2 hours, after which the saturated ammonium carbonate solution (2 L) and toluene (4 L) were added and shaken for 5 minutes. The phases were separated and the aqueous layer was extracted with benzene (2 X 4 L). The mixed toluene extract was washed with diluted hydrogen acid until the solution became homogeneous and washed with brine (3 X 4 L). The solution was finally concentrated to 2 L under atmospheric pressure. This solution was used directly for the next step. • Line - Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printed synthetic (2R,S)-7-hydroxy-3-(4'-hydroxyphenyl)-4-methyl-2-(4"·ρ," Hydropyridine [ethoxy]phenyl)-2Η-1 -benzoquinone (IS)-10-camphorsulfonate (earth 12). 4-(4-[4-[2-~-hexafluoroantimony] ethoxy]-stupyl- 3·(4''-tetrahydropyranyloxy) -7-Tetrahydrogen is added to the toluene solution of chromane (10) in acetone (6 L), water (〇·3 L) and (S)10-camphoric acid (561 g, 2.42). Mole) (available from Aldrich Chemical Company Inc., Milwaukee, Wis.). The mixture was shaken under nitrogen for 48 hours to be solid (2R 'S)-7 -transyl-3-(4, phenyl)-4-methyl-2(4"·[2", hexahydro After acridine]ethoxy)phenyl)-2-indole-1-indole (1S)-10-camphorsulfonate (12) was filtered, washed with acetone and dried (883 g). This material will be used in the next step (HH) without further purification. Synthesis of (2S)-7-carbyl-3-(4'-pyridylphenyl)-4-methyl-2-(4''-[2''' six-paper scale for Chinese National Standard (CNS) A4 size (210 X 297 mm) 60 Economic θΓIntellectual Property Bureau Staff Consumer Cooperative Printed 1293251 V. Invention Description (57 Hydrogen Acridine] Ethoxy) Phenyl)·2Η·1·Benzene π Ratio (ι§) -1 (μ樟 樟 酸盐 ( (13, (+)-£1^1-652 (18)-08 八 salt). (211,8) -7-class _ base -3- (4' Base)_4·methyl-2-(4''-[2'''-hexahydroacridine]ethoxy) stupyl)_2Η-stupid (ls)_1〇_ suspension of camphor sulfonate ± 12 (759 g) was heated in 95% ethanol to about 70 ° C until the solids dissolved. The solution can be stirred and cooled to room temperature, after which some (2S)-7-hydroxy-3-(4,-hydroxyphenyl)4-methyl-2-(4-[2-hexahydropurine] is added. Crystallization of ethoxy)phenyl)-2Η-1·benzopyrenepyranyl sulphate 13 salt. The solution was stirred at room temperature for about 3 days. The filtered crystallization was washed with 95% ethanol and dried (291 g, 76%). The product was removed by 94.2% and the purity was 98.8%. Synthesis of (S)-(+)-7-hydroxy-3-(4'-hydroxyphenyl)-4-methyl-2-(4,,_(2,,, hexahydroacridinyloxy)phenyl Hydrogenated em_〇1538 (EM-652, HC1). Compound π (EM-652-(+)-CSA salt, 500 mg, 〇·726 mmol) in dimethylformaldehyde (11, 〇15 mm The suspension in 〇1) was treated with 0.55 aqueous sodium carbonate solution (7·〇mL, 3.6 mmol) and mixed for 15 minutes. The suspension was treated with ethyl acetate 〇mL) and mixed for 4 hours. The organic phase was then washed with aqueous saturated sodium carbonate solution (2×5 mL) and brine (1×5 mL) dried over magnesium sulfate and concentrated. The pink foam solution (EM_652) produced in ethanol mL) was treated with 2n hydrochloric acid (4 〇〇, 〇·80 mmol) and stirred! After an hour, it was treated with distilled water (5 mL) and stirred for 30 minutes. The resulting suspension was filtered, washed with distilled water (5 mL), dried in air and applied to a high-vacuum (65:[:) to give a milky powder (276 mg 77 / 〇). Fine white powder, scanning Cal〇rimetry): The melting point peak starts at 219t:, ΔΗ = 83 J/g, · U]24d = This paper scale is suitable for fiscal standards (CNSM4 specification (2) G χ 297 mil) ^----- ---^---------Line (please read the notes on the back and fill out this page) 61 1293251 A7 _B7 _ V. Invention description (58 ) (Please read the notes on the back and fill in the form) Page) 154. At 10 mg/ml methanol; β NMR (300 MHz, CD3OD) (5 (ppm) 1.6 (width, 2H, H-4"'), 1.85 (width, 4H, H-3" and 5"), 2.03 (s , 3H, CH3), 3.0 and 3.45 (width, 4H, H-2, ", and 6"), 3.47 (t, J = 4.9 Hz, 2H, H-3",), 4.26 (t , J=4.9 Hz, 2H, H-2”,), 5.82 (s, 1H, H-2), 6-10 (d, J=2.3 Hz, 1H, H-8), 6.35 (dd, J= 8.4, 2·43 Hz, 1H, H-6), 6.70 (d, J=8.6 Hz, 2H, H-3' and H-5'), 6.83 (d, J=8.7 Hz, 2H, H-3 "and H5"), 7.01 (d, J = 8.5 Hz, 2H, H-2, and H-6,), 7.12 (d, J = 8.4 Hz, 1H, H-5), 7.24 (d, J = 8.6 Hz, 2H, H-2" and H-6"); 13C RMN (CD3OD, 75 MHz) (5 ppm 14.84, 22.50, 23.99, 54.78, 57.03, 62.97, 81.22, 104.38, 109.U, 115.35, 116.01 ' 11868 ' 125.78,126.33,130.26,130.72 » 131.29 » 131.59 ,134.26,154.42,157.56,158.96,159.33. Elemental composition: C,Η,N,Cl: theory; 70.51,6·53,2·84,7.18 , %, found: 70.31, 6.75, 2.65, 6.89%. Example 4 Materials and Methods Animals Female BALB/c mice (BALB/cAnNCrlBR) with a weight of 18-20g printed by the Intellectual Property Office of the Ministry of Intellectual Property of the Ministry of Finance were obtained from Charles-River, Inc. (St-Constant, Quebec, Canada) and per cage. 5 are in the sputum, in a controlled environment with temperature (23 ± Γ [:) and illumination (12 hours per illuminating, 7: 15 illuminating). These mice have no rodent chewing grass and faucet water. Restricted supply of food. These animals were removed from the ovaries (usvχ) via bilateral flank incision under Isoflurane anesthesia, and the paper size applies to the Chinese National Standard (CNS) A4 specification (210 X 297 public) 62 ^济郎智慧财 3. The leader hi consumption ^作?!^-代代1293251 Α7 -------- Β7 5, invention description (59) and randomly assigned to each group of 10 animals. Another 10 mice remained intact as a control group. Treated in the first batch of experiments (Figures 12 to 15), test compounds, namely EM-652.HC1, Lassf〇xifene (as free test; activated and deactivated mirror image) And Ral〇xifene, oral administration of each sturdy silver-human 'inscribed as 1, 3 or 1 〇 # g / animals began to treat 9 于 after 2 ovariectomy. In the second batch of experiments (Table 3), TSE 424 was administered orally at a dose of 1, 3, 10 or 30 and the animal was started on the 9th day after ovariectomy. In the two batches of experiments, in order to evaluate the activity of antiestrogens, the female pigment (El, 0·06 #g, s.c. injection, twice daily) was treated on the 5th day after the ovarian knife was removed, and the drug was administered for 6 days. The compound was dissolved in ethanol (final concentration 4 〇/〇) and administered in 〇·4% methylcellulose. The mice in the intact and 〇νχ control groups received vehicle alone (4% ΕΤ〇Η-〇4% methylcellulose) at /9 曰 on the first morning of ovariectomy, and the animals were bled with abdominal aorta. The way to kill. The uterus and sheath are rapidly dissected, weighed and stored in 10% buffered formalin for further tissue testing. RESULTS Experiment 1: As shown in Figure 12, EM-652 Hc was administered at a daily oral dose of μL, 34 ^ and administered, respectively, resulting in 24%, gamma, and 72% inhibition of uterine weight in females (all dose pairs) The control group, ρ < 〇〇ι), and Raloxifene administered at the same dose resulted in 6% (NS), 14% (".01) and 43% (ρ<〇·〇) for this parameter, respectively. The inhibition of ι). In addition, ------------- loading -------- order --------- line (please read the notes on the back and fill out this page)

63 1293251 A7

V. INSTRUCTIONS (60) The Ministry of Economic Affairs' Intellectual Property Office employee consumption cooperative printed π Lassofosif (LaS0f0xifene) (as a free base) did not have an inhibitory effect at the lowest dose, but it used a dose of 3 at each dose. The temperate resulted in inhibition of 25% (ρ<〇〇ι) and 44% (ρ<0·01) of the female stimulating uterine weight, respectively. The deactivated mirror of the lassofufene does not inhibit this parameter at any dose. The above compounds produced a similar effect on the weight of the sheath. Each oral dose of EM-652.HC1 resulted in 10% (NS), 25%, and 53% inhibition of sheath weight at 1 g, 3 V g, and 10 // g dose (Fig. 13), respectively. 2 In the higher dose, 'Ρ<〇·〇1), and Raloxifene only produces a 24% (ρ<〇·〇ΐ) meaningful inhibition on this parameter at the highest dose. Similar to Raloxifene, Lassf〇xifene (as a free test) causes only a significant inhibition of 37 〇/〇 (p<〇〇1) at the highest dose used, Deactivation of the mirror image did not inhibit sheath weight at any dose. EM-652.HC1 does not have uterine weight at 2 doses when the compounds are administered alone (without female) to the ovariectomized mice at daily oral doses of 1 " g and 1 〇 #g. Significant stimulating effects, resulting in 93% (ρ<0·01) and 85% (ρ<0·01) uterus, respectively, when treated with 1〇" g Lasosofoxifene and Raloxifene Stimulation of weight (Fig. 14), thus showing the estrogenic effects of these compounds on this parameter. Similarly, EM-652.HC1 did not have a significant inhibitory effect on sheath weight (Fig. 15), while 1〇# g Lassofxxifene and Raloxifene administered 73%, respectively. Sheath weight stimulation of (ρ<〇·〇ι) and 56/〇(ρ<〇〇1). On the other hand, the paper size of the Lasso Fofifen applies to the Chinese National Standard (CNS) A4 specification (2) 297 297 mm) (please read the notes on the back and fill out this page). --Line· 64 1293251 V. INSTRUCTIONS (61 (Lasofoxifene) deactivated mirror image did not have a significant inhibitory effect on uterine and sheath weight. Experiment 2: As shown in Table 3, TSE 424 was given 1// § per dose, 3"g, 1〇# g or 30/zg administration resulted in 12% (NS), 47%, 74%, and 94% female hormones stimulating uterine weight inhibition. Inhibition of female stimulating uterus weight (3 highest doses) For the El• control group, p<〇〇1). On the other hand, daily oral administration of 3 // g, 1〇# g, and 30/zg doses of D8 £ 424 resulted in (NS), 56% (鞘<0·01) and 93% (p<〇〇1) for inhibition of sheath weight. When these compounds are administered at a dose of § and 3 〇// per dose per dose (excluding female) to small oophorectomy In rats, TSE 424 did not have a significant stimulating effect on uterine and sheath weight at both doses (Table 3). — (Please read the notes on the back and refill Write this page) Ministry of Economic Affairs Intellectual Property Bureau employees consumption cooperation 钍 printed table 3 · The effect of y-concentrated TSE on the uterus and sheath weight of oral ovarian resection mice treated with or without treatment **Ρ<〇·〇1, for the control group treated with El. Treatment of uterine weight (mg) Sheath weight (mg) Complete 54·6±12·5** 37·9±3·9" OVX 15.6±1·3" 13·9±1·5** OVX + E1 118.3±6·0 53·4±2·8 OVX + El + TSE 424 1 // g 105.5±6.1 54.2±3.0 OVX + El + TSE 424 3 // g 69.7±4.4** "47.2±1·6 — OVX + E1 + TSE424 10# g 42.1 ±2.7** 31.1±2·3" OVX + El + TSE 424 30// g 21.7±1 ·7** 16·7±1·8” ~ OVX + TSE 424 3 /zg 18.3±1·2 14·1±1·2 OVX + TSE 424 30 // g 17.7±1.6 1 5·3 ±2· 0 -- Line · This paper scale applies to China National Standard (CNS) A4 specification (210 X 297 mm) 65 1293251 a; B7 Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printing 5, invention description (62) Example 5 5 A · Preventive effects on bone loss, serum lipids and total lipids. Animals and Treatment Female Spock-Dora rats (Crl: CD (SD) Br) (Charles River Laboratory, St-Constant, Canada) weighing 10 to 12 weeks old and weighing about 220-270 g at the start of treatment were used. Before starting this experiment, the animals were first adapted to the environmental conditions (temperature: 22 ± 3 ° C; humidity: 50 ± 20%; 12 hours of light - 12 hours dark cycle, starting at 07 · 15 hours) at least 1 week. The animals were individually closed and allowed free access to faucet water and approved granular rodent feed (Lab Diet 5002, Ralston Purina, St-Louis, MO). The experiment was conducted in the animal equipment approved by the Canadian Council on Animal Care (CCAC) and the Association for Assessment and Accreditation of Laboratory Animal Care (AAALAC) according to the CCAC Guide for Care and Use of Experimantal Animals manual. 154 rats were randomly assigned to a cohort of 14 animals per group as follows: 1) intact control group; 2) OVX control group; 3) OVX+E2 (lmg/kg); 4) OVX + EM-652 .HC1 (2.5 mg/kg); 5) OVX + E2 + EM652.HC1; 6) OVX + dehydroepiandrosterone (DHEA; 80 mg/kg); 7) OVX + DHEA + EM-652.HC1; OVX + DHEA + E2 ; 9) OVX + DHEA + E2+ EM-652.HC1 ; 10) OVX + GW 5638 ; 11) OVX + E2 + GW 5638. On the first day of the study, appropriate groups of animals underwent bilateral oophorectomy (OVX) under Isoflurane anesthesia. DHEA is dissolved in 50% ethanol-50% propylene glycol (please read the back of the note before you fill out this page) Order · · • Line -

-T The paper size applies to the Chinese National Standard (CNS) A4 specification (210 X 297 mm). 66 3⁄4 齐郎 are all yc inspectors X-Shaw fee, & v乍i. 1293251 A7 ~------ --B7__ V. INSTRUCTION DESCRIPTION (63) The liquid was applied topically to the dorsal epidermis, while the other test compounds were suspended in 0.4% methylcellulose for oral administration. Treatment started at 苐2 本 of the study and was processed once a day for 3 months. In the second batch of experiments, 132 rats were randomly assigned to 9 groups of 14 or 15 animals per group, as follows: 1) intact control group; 2) 〇VX control group; 3) 0VX+Pima Forest (Premarin) (0.25 mg/kg); 4) 0VX + ^ EM-652.HC1 (2.5mg/kg); 5) OVX+pimarin (?11^1^11) + £]\4- 652 .HC1 ; 6) OVX + TSE 424 (2.5 mg/kg); 7) OVX + Pimarin (Premarin) + TSE 424; 8) OVX + Lassofoxifene (tartrate; racemate; 2.5 mg /kg) ; 9) OVX+Premarin + Lasofyfoxifene. On the 1st day of the study, the appropriate group of animals underwent bilateral ovariectomy (OVX) under Isoflurane anesthesia. The test compound was suspended in 0.4% of thioglycolic cellulose and administered at a dose of sputum. Treatment began on the second day of the study and was processed once per week for 26 weeks. In the 2 batches of experiments, animals that did not receive the test substance were treated with the appropriate vehicle alone in the same period. Bone mineral density measurement ^ 3 months (Experiment 1) or 26 weeks (Experiment 2) treatment, individual rats under the Isoflurane anesthesia, the whole body bone and lumbar spine receive double energy X-ray absorbance analysis (DEXA; QDR 4500A, Hologic, Waltham, ΜA) was scanned and Regional High Resolution Scan software was used. Determine the bone mineral density (BMD) and overall composition (percentage of fat) of the lumbar spine (vertebra L2 to L4). Serum test The paper size applies to the Chinese National Standard (CNS) A4 specification (210 X 297 mm) --------^. — —---- (Please read the note on the back and fill out this page) 67 1293251 A7 ---- B7 V. INSTRUCTIONS (64) After 3 months (Experiment 1) or 26 weeks (Experiment 2), blood samples were collected from the jugular vein of an animal that had not been fed overnight (in Aesop Fran (IS0flurane) under anesthesia). Samples were processed to facilitate serum preparation and refrigerated at -80 until testing. Serum cholesterol levels and lyserase activity (ALP) were determined using a Boehringer Mannheim Diagnostic Hitachi 911 Analyzer (Boehringer Mannheim Diagnostic Laboratory Systems). Statistical analysis Data are expressed as mean soil SEM. Statistical significance was determined according to the multi-range test of Dimcan-Kramer (Kramer CY; Biometric 1956; 12: 307-3 10). Results As shown in Table 4, after 3 months of ovariectomy, the BMD of the lumbar spine was lower than that of the intact control group by ι〇〇/0 (p<〇1) in the OVX control animals. At the doses used, estradiol and EM-652.HC1 alone prevented the loss of lumbar spine BMD by 98% (ρ<〇·〇1) and 65% (ρ<〇·〇5), respectively, and Ε2 In combination with EM-652.HC1, the reduction in BMD reduction in the lumbar spine caused by OVX was reduced by 61% (ρ<0.05). On the other hand, although DHEA alone can avoid the lumbar spine BMD by 43% (ρ<〇.〇5), the combined treatment with DHEA+ Ε2 + EM-652.HC1 avoids the ονχ-induced lumbar spine BMD reduction of 91 %, and caused its BMD value to be no different from the intact control group. In Table 5, the BMD of the lumbar spine was reduced by 18% (p < 0·01) compared with the intact pair and 26 weeks after ovariectomy. Preniarin, EM-652.HC1, TSE 424 and Lasofofifene are administered separately. The paper size applies to the Chinese National Standard (CNS) A4 specification (210 X 297 mm). (Please read first. On the back of the note, please fill out this page. -# - Line - Ministry of Economic Affairs, Intellectual Property Bureau, Staff and Consumer Cooperatives Printed 68 1293251 A7 B7 V. Inventions (65) (Please read the back note and fill in the form) Page) Avoid reduction of lumbar spine BMD by 54%, 62%, 49%, and 61%, respectively (all ρ < 0·01, vs. OVX control group). The addition of Pimalin (Premarin) to Ugly River-652.HC1, TSE 424 or Lasofyfoxifene resulted in no significant difference in the BMD values of lumbar spine and individual SERM alone (Table 5). Similarly, the addition of DHEA to E2 or to EM-652.HC1 completely avoided LVX-induced reduction in lumbar spine BMD (Table 4). The positive effect of DHEA-BMD is also supported by its role in serum alkaline phosphatase activity (ALP), an indicator of bone formation and transformation. ALP activity increased from 73 um 6 IU/L in OVX control animals to 224 ± 18 IU in DHEA, DHEA + EM-652. HC Bu DHEA + E2 and DHEA + E2 + EM-652. HC1 treated animals. L, 290 ± 27 IU / L, 123 ± 8 IU / L and 261 ± 20 IU / L (all p < 0.01), thus showing the stimulating effect of DHEA on bone formation (Table 6). In addition to avoiding bone loss, administration of EM-652.HCn, TSE 424, Lasofyfoxifene, GW 5638, DHEA, and E2 produced some beneficial effects on overall fat percentage and serum lipids. Three months after ovarian resection, the overall fat increased by 22% (ρ<0·05; Table 6). Ministry of Economic Affairs, Intellectual Property Bureau, Staff Consumer Cooperative, Printed Clothes ~ EM-652.HC1 administration completely avoids the increase in fat percentage caused by OVX, while adding DHEA and/or £2 to SERM results in lower fat percentage values than in intact control animals. The measured value. After 26 weeks of oophorectomy, the 40% increase in fat caused by estrogen deficiency returned 74% after administration of Premarin, EM-652.HC1, TSE 424 or Lasoffoxifene. 78%, 75%, and 114%, and the addition of Pimarin (?1^11^1^11) to each SERM completely avoided the increase in the percentage of fat caused by OVX (Table 7). This paper scale applies to the Chinese National Standard (CNS). A4 size (210 X 297 mm) 69 A7 1293251 _B7_ V. Description of invention (66) 0 As shown in Table 6, after 3 months of ovariectomy, serum cholesterol levels were observed in OVX control rats. The control group (ρ<〇.〇1) increased by 22%. In fact, serum cholesterol increased from 2.01 ± 0.11 mmol/L in intact animals to 2.46 ± 0·08 mmol/L in the OVX control group. Ε2 or DHEA alone reduced serum cholesterol levels to 1.37 ± 0.1 8 mmol/L and 1.59 ± 0.10 mmol/L, respectively, whereas EM-652.HC1 alone or in combination with £2 and/or DHEA resulted in cholesterol levels. The level found in intact animals (2.01 ± 0.11 mmol/L) decreased significantly (between 0.65 and 0.96 mmol/L). Similarly, GW 563 8, TSE 424, and Lasofofoxifene alone or in combination with E2 or Premarin completely avoided the increase in serum cholesterol levels induced by OVX and resulted in a higher value. The value of intact animals was low (Tables 6 and 7). (Please read the notes on the back and fill out this page) - Line · Ministry of Economic Affairs Intellectual Property Bureau Employees Consumption Cooperatives Printed on paper The paper size applies to National Standard (CNS) A4 (210 X 297 mm) 70 1293251 A7 B7 V. INSTRUCTIONS (67) Table 4: Treatment of ovariectomized rats with estradiol, EM-652.HCn, GW 5638 or DHEA to avoid bone loss after 3 months, administration of sputum alone or in combination Γ Treatment of lumbar spine BMD (g/cm2) Avoid bone flow f (%) Complete 0.2461 ±0.0049** Too" OVX 0·2214±0·0044 • ovx + e2 0·2457±0·0049** ~98 ^ OVX + EM-652 .HC1 0.2374±0.0027* 65^~ OVX + EM-652.HC1 +E2 0.2364±0.0037* OVX + DHEA 0.2321 ±0.0034 OVX + DHEA + EM-652.HC1 0.2458±0.0037** ^99 ~ OVX + DHEA + E2 0·2496±0·0029** OVX + DHEA + E2 + EM-652.HC1 0·2439±0·0043** OVX + GW 5638 0.2299±0·0060 OVX + GW 5638 + E2 0.2344±0.0054 ,ρ<0·05; **, p < 0.01, experimental group relative to 〇VX control group of rats Table 5: PREMARIN, EM-652.HCM, TSE 424 or Lassofoxifene deal with After 26 weeks of ovarian resection of female rats, the effect of avoiding the loss of bone thief, alone or in combination with pseudomalin (PREMARIN). Bay-------------- Pack---C Read the following notes on the back and then fill out this page> 2^1. ) Lumbar Spine Treatment - Line · BMD (g/cm2) Ministry of Economic Affairs Intellectual Property Bureau Staff Consumption Cooperation Du Yin ^4 Complete ΟΫΧ 0·2482±0·0067 ** 0.2035±0.0035 100 OVX+Pimarin ovx +~EM^52.HC1 OVX +Pimarin + EM-652.HC1 〇VXTlSE424~- 〇VX+Pimarin +TSE 424 〇vx ^ 5vx 十互石^拉蚤佛西芬0·2277±0·0028** 0.2311 ±0.0040** 0·2319±0.0057** 0·2252±0·0058** 0.2223 ±0.0046** 0.2307±0.0040** 0·2357±0· 0035** Ρ<0·(Η 'The experimental group is relative to the 〇νχ control group. This paper scale applies the Chinese national standard (CNS) A4 specification (2) 0 x 297 public)

71 A7 1293251 __ B7 V. INSTRUCTIONS (68) Table 6: Total fat percentage, serum cholesterol level and after 3 months of ovariectomy in female rats treated with estradiol, EM-652.HC1, GW 5638 or DHEA The effect of alkaline phosphatase activity, administered alone or in combination. Total fat (%) Cholesterol (mmol/L) ALP (IU/L) Complete 24·0±1·5* 2·01±0·11** 39±2** OVX 29·2±1·5 2 ·46±0·08 73±6 ovx+e2 19·5±2·5** 1.37±0·18** 59±4 OVX+EM-652.HC1 23·2±1·4** 0.87+0.04 ** 91±6* 0VX+EM-652.HC1+E2 20.4±1.4** 0·96±0·07** 92±5* OVX+DHEA 17·3±1.5** 1.59+0.10** 224± 18** OVX+DHEA+EM-652.HC 1 18·0±1.1** 0.65±0·06" 290±27** ovx+dhea+e2 15·8±1.3** 1·08±0·08 ** 123±8** OVX+DHEA+E, +EM- 652.HC1 19.2±1·6** 0.71 ±0.08** 261 ±20** OVX+GW 5638 21.9±1.4** 1·14±0.08 ** 72 ±6 OVX+GW 5638+E2 23.2+1.2** 0.91+0.07** 80 + 6 ,ρ<0·05 ; **,ρ<0·(Μ, experimental group relative to OVX control rats (Please read the notes on the back and fill out this page) -J. - . Table 7: Treating nests with PREMARIN, EM-652.HQ, TSE 424 or Lasoffoxifene Effects of total female fat percentage, serum cholesterol level and alkaline phosphatase activity on female rats after 26 weeks of excision, either alone or in combination with pseudomalin (PREMARIN). Property Bureau Staff Consumer Cooperative Printed Total Fat (%) Cholesterol (mmol/L) ALP (IU/L) Complete 25·5±1·8** 2·11±0.11** 33±2* OVX ' 35· 7±1·6 2.51 ±0.09 60±6 OVX+Pimarin 28·2±1·8** 1·22±0·07** 49±3 OVX+EM-652.HC1 27·7 士1.4** 0·98±0·06** 78±4 OVX+EM-652.HC1+pimarin 25.7±2.2** 1.10±0.07** 81±6 OVX+TSE424 28·0±1.8** 1.15±0.05** 85±6 OVX+TSE424+pimarin 25·7±1·7** 1·26±0·14** 98±22** OVX+ 拉索佛西芬 OVX + 拉索佛西芬+皮马林24 ·1±1·3** 23·8±1·9"0·60±0.02" 〇·81±0·12** 116±9** 107±6** P<0_05; **, ρ&lt ;0·01, the experimental group was compared to the 〇νχ control group. This paper scale applies to China National Standard (CNS) A4 specification (210 X 297 public interest) 72 1293251 A7 B7 Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printed invention description (69) 5B: treatment of bone loss and overall fat effect Female Spock-Doray rats (Crl: CD(SD)Br) (Charles River Laboratory, St-Constant, 8 to 9 months old) were used in this experiment.

Canada). Prior to ovariectomy, the animals were first acclimated to ambient conditions (temperature: 22 ± 3 ° C; humidity: 50 ± 20%; 12 hours of light - 12 hours dark cycle, starting at 07: 15) for at least 1 week. The animals were resected bilateral ovaries (0VX) under anesthesia with Isoflurane. Twenty animals were maintained intact as a control group. The animals were individually closed and allowed free access to faucet water and approved granular rodent feed (Lab Diet 5002, Ralston Purina, St-Louis, MO). The experiment was conducted in the animal equipment approved by the Canadian Council on Animal Care (CCAC) and the Association for Assessment and Accreditation of Laboratory Animal Care (AAALAC). According to the CCAC Guide for Care and Use of Experimental Animals, 10 weeks after 0 OVX, 139 Rats were randomly assigned to groups of 17 to 20 animals per group as follows: 1) intact control group; 2) OVX control group; 3) OVX + E2 (1 mg/kg); 4) OVX-fEM- 652.HC1 (2.5 mg/kg); 5) OVX+ E2+EM-652.HC1; 6) OVX+ dehydroepiandrosterone (DHEA; 80 mg/kg); 7) OVX DHEA+EM-652.HC1; 8) OVX+DHEA + EM-652.HC1+ E2. DHEA was applied topically to the dorsal epidermis in a solution of 50% ethanol-50% propylene glycol, while the other test compounds were suspended in 0.4% methylcellulose for oral dose administration. Treatment begins after the first week of oophorectomy and once daily for 26 weeks. The test papers are not accepted. The Chinese National Standard (CNS) A4 specification (210 X 297 mm) is applied to the paper scale. -------------Installation------ -----Line (please read the note on the back and then fill out this page) 73 1293251 A7 Ministry of Economic Affairs Intellectual Property Office Staff Consumer Cooperative Print π B7_ V. Invention Description (7〇) Animals in the same period The appropriate carrier is treated separately. Bone Mineral Density Measurement Individual rats underwent dual-energy X-ray absorptiometry (DEXA; QDR 4500A, Hologic, Waltham, MA) and were used under the anesthesia of Isoflurane for dual energy X-ray absorptiometry (DEXA; QDR 4500A, Hologic, Waltham, MA) Regional High Resolution Scan software. Determine the bone mineral density (BMD) and overall composition (percentage of fat) of the lumbar spine (vertebra L2 to L4). Statistical analysis Data are expressed as mean soil SEM. Statistical significance was determined according to the multi-range test of Duncan-Kramer (Kramer CY Biometric 1956; 12: 307-3 10). Results In the previous study (Example 5A), in order to learn the role of preventing bone loss, administration of the test compound was started at the time of OVX. In order to study the possible therapeutic effects of the drug administration, in this study, the administration of the test compound was started 10 weeks after OVX. In order to establish the presence of defects in bone formation prior to the start of treatment, BMD was measured (reference value) before OVX and treatment started. As shown in Figure 16, the BMD of the lumbar spine was reduced by 8% after 10 weeks of oophorectomy and further reduced by 12% during the other 26 weeks after OVX, where the animals were only individually received during the 26 weeks. Agent (control group). Daily administration of E2 'EM-652.HC1, E2+EM-652.HC1, DHEA or DHEA+EM-652.HC1 to animals with established osteogenic defects was completely avoided in OVX control animals for 26 weeks. The paper size is applicable to the Chinese National Standard (CNS) A4 specification (210 X 297 mm) (please read the notes on the back and fill out this page) · -1 line · J · 74 1293251 _ B7___ V. Description of invention ( W) (Please read the note on the back and then fill out this page) The BMD of the one-step lumbar spine is reduced, while the E2+EM-652.HC1+DHEA administration causes the BMD value to rise slightly compared to the value observed before the start of treatment. On the other hand, as exemplified in Fig. 17, femoral BMD decreased by 4% after 10 weeks of oophorectomy, and further decreased by 6% after another 26 weeks of treatment of the vehicle alone. Similar to lumbar spine BMD, daily administration of E2, EM-652.HC1, a E2+EM-652.HC1, DHEA or DHEA+EM-652.HC1 26 weeks completely " avoided in OVX control animals The further lumbar spine to the age of BMD decreased. On the other hand, administration of E2+EM-652.HC1+DHEA resulted in a slight increase in the BMD value of the femur even compared to the value observed before OVX, thus showing the beneficial effect of the combination treatment on bone formation. The combined treatment of EM-652.HC1+E2+DHEA not only completely prevented OVX from triggering further bone loss in osteoid-deficient animals, but also produced some therapeutic effects. In addition to its effect on bone, as illustrated in Figure 18, DHE A, E2*, or EM-652.HC1 administration avoids the overall fat increase induced by OVX >. In fact, in the OVX control animals, the percentage of fat increased by 47% after 10 weeks of printing in the Ovarian Cut Economics Department of Intellectual Property, and in the treatment of vehicle alone (control group 'group) In the 26 weeks, it was further reduced by 17%. Daily administration of E2, DHEA, EM-652.HC1, E2+EM-652.HC1 or DHEA+EM-652.HC1 26 weeks avoided the 17% increase observed in the OVX control group. On the other hand, the combination treatment EM-652.HC1+E2+DHEA completely restored the effect of OVX and caused the fat percentage value to be similar to that found before the ovariectomy in the animal, thus showing the therapeutic effect of the treatment. . This paper scale applies to China National Standard (CNS) A4 specification (21u X 297 mm). 75 A7 1293251 _____BT^___ V. Invention description (72) Example 6 (Please read the note on the back and fill out this page) Compound of the invention Effect of alkaline phosphatase activity on adenocarcinoma Ischwaca cells in human endometrium. Materials Maintenance of cultured cells The human Ischwarka cell line derived from a well-differentiated endometrial adenocarcinoma was presented by Dr. Erlio Gurpide, The Mount Sinai Medical Center, New York, NY. Ischwara cells are routinely cultured in Eagle's Minimum Essential Medium (MEM) containing 5% (vol/vol) FBS (fetal bovine blood _ clear) supplemented with 100 U/ml penicillin, 100 g/ml streptavidin And 0.1 mM non-essential amino acid solution. At 37 ° C, the cells were seeded at a density of 1·5 X 106 on the bottom of a Falcon T75 conical flask. Cell Culture Experiment--Line-Economics Bureau of the Intellectual Property Office of the Ministry of Economic Affairs, 24 hours before the start of the experiment, the culture medium of the fusion of Ischwaka cells was replaced with fresh estrogen-free basic culture solution (EFBM). Base medium consisting of 1: 1 (v: v) phenol red-free Ham's F-12 and supplemented with 100 U/mL penicillin, 100# g /mL streptomycin, 2 mM amidoxime and 5% FBS Dulbecco's Modified Eagle, s Medium (DMEM) consists of 'treated with xylogan coated wood carbon twice to remove endogenous steroids. The cells were harvested with 0.1% trypsin (Sigma) and 0.25 mM HEPES, resuspended in EFBM and plated at a volume of 100 x / 1 in a Falcon 96-well flat-bottomed microtiter plate at 2.2 x 104 cells/well, and allowed to attach to the disk surface. 24 hours. Thereafter, the fresh EFBM medium containing the indicated concentration of compound was replaced with a final volume of 200 ul. Cell culture for 5 days, every 48 hours. This paper scale applies to China National Standard (CNS) A4 specification (210 X 297 mm) 76 1293251

V. Description of the invention (73) Culture medium. Test for luciferase At the end of the culture period, the microtiter plate is inverted and the growth medium is allowed to flow out slowly. Each well of the plate was wetted with 200//1 PBS (0.15 M NaCh 10 mM sodium phosphate 'PH 7.4). The Pbs were then removed from the disk and some residual PBS was carefully left, and the cleaning procedure was repeated several times. Slowly pour out the buffered saline solution and blot the inverted plate gently with a paper towel. After replacing the lid, the tray was placed at -80 ° C for 15 minutes and then allowed to melt at room temperature for 1 minute. The plate was then placed on ice and ice-cold solution containing 5 mM p-nitrate, 0.24mMMgCl2 and 1 Μdiethanolamine (ρΗ9·8) 5〇ν1 was added. The plate was then warmed to room temperature and allowed to develop a yellow development (8 minutes) of the ρ. The change at 405 nm was monitored in an enzyme-linked immunosorbent assay (81 〇 RAD, model 2550 EIA Reader). The calculated dose response curve and IC5G values were calculated using a weighted replicated nonlinear squared regression method. -------------Install--------Book---------Line (Please read the back of the matter and then fill out this page)

Ministry of Economic Affairs, Intellectual Property Bureau, Employees, Consumers, Cooperation, Printing, Printed Paper, Applicable to China National Standard (CNS) A4 Specification (21ϋ X 297 mm) 77 Ministry of Economic Affairs, Intellectual Property Office, Staff and Consumer Cooperation, Printed Agriculture, 1293251 V. Invention Description (74 ) 〇U) 00 〇3: 辫m 2 1 On LT\ to X n W h_^ 2 1 00 00 〇^ w m2 2 & ^ i° u) 3C ^ G Taili-vP 〇〇k τ&gt ; k Ό miscellaneous ^1+ U\ L/i κ> )0 0'S 3 1+ ΙΟ I—k 1—* bo ^ oo β 1+ w 〇k) ON sll 5 z 5 @ 3严ϋ Κ) One two ^ 1+ Ον 1/1 K) S 1+ w 〇to K) G photo 1 ^ ^ xj SS /^ -f 0 4 八ρί /^S Lk) S 1+ LO v〇OO /—V 】 =丨+ wp α lifting s |^ s ^ p^ — 棼> edge---------/9·---L----- (please read the notes on the back and fill out this page) 1]· · · Green - This paper scale applies to China National Standard (CNS) A4 specification (210 X 297 mm) 78 1293251 V. Description of invention (75)

Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printed ΙπΜ E2 窆 八8*%=00 405 nM ;& 碎 /0D 405 nM^i*蜂 and Labrie46A EM — 642(SCH 5 7068)-1 黹铖w^'芩»IL abuse»^^2^^^^^^#^-t^ssEIlMJ.SteroidBiochem.AndMO.Bi?69,5rs,1999 m # X U3> s- >W/ -< U) U) Lk) 00 Book r Η—* Lh ON 00 蓉 m 2 1 L/\ to m 2 ΰ> m 2 1 η—^ guest m 2 K η—^ ^>mk 黎Ο \ X ^ o? CT ^ ο 〇\ w Ss & NJ ^ bo 5 1+ N-^ 二芸HA bo O L> S 1+ ov〇Μ % -3* ^ tk Q§ w5; 00 a k) Ss L) /-s ·— * S 1+ # % δ s ------------- loading -------- order --------- line (please read the precautions on the back) Fill in this page) This paper scale applies to China National Standard (CNS) A4 specification (210 X 297 mm) 79 1293251 A7 Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printing 5, invention description (76) Example 7EM-652.HC1 Effects of TSE 424 and ias〇f〇xifene on proliferation of human breast cancer MCF-7 cells. Method: Maintenance of cultured cells MCF-7 Human breast cancer cells The 147th generation line was obtained from American Type Culture Collection #HTB 22 and was routinely cultured in Dulbecoo's Modified Eagle's-Ham's F12 medium without redness. The supplement and 5% FBS. The MCF-7 human breast cancer cell line was derived from pleural effusion of a 69-year-old Caucasian female patient. The MCF-7 cell line used was between passages 148 and 165 and subculture was completed weekly. After the cell proliferation study, the cells in the logarithmic growth phase were collected in 〇·1% digester (Sigma), resuspended in a suitable medium containing 50 ng of bovine insulin/mi, and 5 〇/〇 (v/v) FBS and The glucomannan coated carbon was treated twice to remove the endogenous solid bile. The cells were seeded in 24-well {^1 (: 011 plastic plates (2 cmWL) at the indicated density and allowed to attach to the surface of the plate for 72 hours. Thereafter, the cells were replaced with fresh cultures containing the indicated concentrations of compounds. Liquid, these compounds are obtained by diluting the concentrated storage/serum solution with or without E2 and soluble in 99% of re-distilled ethanol. The control cells only receive the ethanol carrier (〇·丨% Et) 〇H, v/v). The cells are cultured at specific time intervals and the culture medium is changed every 2 or 3 days. The number of cells is determined by measuring the amount of DNA to determine the scale of the test and the statistical analysis of the paper scale. China National Standard (CNS) A4 Specification (210 X 297 public) — -80 - (Please read the back of the note first and then fill out this page) 4. -Λ · -- Line · 1293251 V. Invention Description (77 The dose response curve and ICm values were calculated using weighted replicate nonlinear minimum order power regression. All results were expressed as mean soil SEM. Table 9 Experiment 1 Name code name dna Test compound for maximum stimulation DNA of test compound ψ:卩丨&ΙΓΛ> ί · InME, stimulating %* ------______ IC50 (nM) EM-652.HC1 bM-〇jz.rlCl5 EM-1538 N.S. 0.796 TSE 424 EM-3527 N.S.

-------------- Strong clothes --- f Please read the notes on the back and then fill out this IC Experiment 2 Name code name DNA Test compound stimulating DNAilnMl^^- test compound Yue, 丨InM Ε,% of stimulation* IC50 (nM) EM-652.HC1 EM-652.HC1; ΕΜ-1538 NS 0.205 ' 拉索佛西分(免碱) ΕΜ-3114 NS 0.379 订· -丨线_ Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative print example 8 compared to nude mice, EM-652.HCL, Tamoxifene, Toremifene, Droloxifene, Idoxifen ), the effect of GW-5638 and Raloxifene on the growth of human RZ-75-1 breast tumors. The purpose of this example was to compare the efficacy of EM-652.HC1 and six other oral antiestrogens (SERMS) in the development of well-characterized estrogen-sensitive ZR-75-1 breast cancer xenografts in ovariectomized nude mice. Antagonism. This paper scale applies to China National Standard (CNS) A4 specification (210 X 297 mm) 81 1293251 A7 Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printed B7 ___ V. Invention description (78) Materials and methods ZR-75-1 The cell ZR-75-1 cell line was obtained from the American Type Culture Collection (Rockville, MD) and cultured in PRMI 1640 medium. These cells were replenished with 2 mM L-glutamine, 1 mM sodium pyrophosphate, 100 IU penicillin/ml, 1 mg streptomycin/ml and 10% fetal bovine serum, and cultured at 37 °C. In humid air with 95% air and 5% carbon dioxide. Cells were seeded weekly and collected at 85-95% confluence using 0.083% trypsin/0.3 mM EDTA. Animal and tumor-bearing female female homozygous (nu/nu) Br aymic mice (28 to 42 days old) were obtained from Charles River, Inc. (Saint-Constant, Quebec, Canada). The mice (5 per cage) were housed in cages of vinyl and air-conditioned airflow hoods and kept under pathogenic conditions. The illumination time is 12 hours and 12 hours dark (lighting starts at 07:15). Cage, mat and feed (Agway Pro-Lab R-M-H Diet #4018) are sterilized prior to use. Drinking water is sterilized without restriction. Simultaneous resection of the ovaries (OVX) was performed under Isoflurane anesthesia. At the time of ovariectomy, estradiol (E2) was injected subcutaneously to stimulate the initial tumor growth. The E2 injection is a Silastic tubing (inner diameter: 0.062 inch; outer diameter: 0.095 inch), 1 cm long and containing 0.5 cm of a 1:10 (w/w) mixture of estradiol and cholesterol. Prepared in. One week after oophorectomy, 2×106 ZR-75-1 (93th generation) cells were passed through a solution of 0.1 ml of RPMI-1640 medium + 30% matrigel (please read the back of the note first) Please fill out this page again. -J · · - Line - This paper scale applies to China National Standard (CNS) A4 specification (210 X 297 mm) 82 A7 1293251 __ B7 ___ V. Invention description (79) 2.5 cm long 22 Standard needles were inoculated subcutaneously to the bilateral abdomen of each ovariectomized (0VX) mouse. After 4 weeks, all animal e2 injections were replaced with the same volume of injection containing female pigment (Ei:chol, 1 ··25, w:w). The processing was started after one week of randomization and processing. On the day before the initial treatment, 255 mice with an average area of 24.4 ± 0.4 mm2 (range 5, 7 to 50, 7 mm2) of ZR-75-1 tumors were randomly assigned to 17 cohorts (depending on tumor volume). Each group contained 15 mice (a total of 29 or 30 tumors). The 17 groups included 2 control groups (OVX and OVX+ feminine), 7 groups supplemented with female sputum and treated with anti-estrogen, and 8 other groups received anti-estrogen alone. Thereafter, the female sputum injection was removed from the ovarian resected control group (OVX) and in the group treated with antiestrogens alone. In nine other groups, the female-containing injection was replaced every 6 weeks. EM-652.HC1, Raloxifene, Droloxifene, Idoxifene, and GW 5638 are in the Laboratory of Molecular Endocrinology of the &... CHUL Research Center ) synthesized in the medical and chemical department. It was purchased from Plantex (Netanya, Israel) and the Tomicifene citrate was purchased from Orion (Espoo, Finland). Under the stimulation of female sex, anti-estrogen was administered at a dose of 50 mg (average 2 mg/kg) per day and suspended in 0.2 ml, 0.4% (w/v) methylcellulose. Animals were treated with 200/g (mean 8 mg/kg) antiestrogens once daily via the oral route without female-stimulation. The paper size of the two groups of the control group is applicable to the Chinese National Standard (CNS) A4 specification (210 X 297 mm) ------------- ·--------Line (Please read the back of the note first and then fill out this page) Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printing 83 V. Invention instructions (8〇) Only accept 〇·2 ml separately Carrier. These appropriate concentrations of the anti-hormone suspension solution are formulated once a month, stored at 4 tons and shaken frequently during use. The powder reservoir is stored sealed in 41 (Id〇xifene, Raloxifene, T〇remifene, Gw 5638, Droloxifene) or stored at room temperature (Moses Tumor (Tam〇xifene), EM-652.HC1) 〇 Tumor measurement and cadaver examination recorded 2 vertical diameters, while tumor area (mm2) was calculated using the formula: L/2 XW/2 X 7τ. The area measured at the first day of processing is treated as 1 . Tumor measurement and cadaver examination 屺 Record 2 vertical diameter, and tumor area (mm2) is calculated using the formula: [π χ W/2 X 7ττ.面积i measured the area of 1%% of the Ministry of Economic Affairs, Intellectual Property Bureau, employee consumption cooperatives

After 161 days of treatment, the remaining animals were anesthetized with Is〇flurane, which was killed by exsanguination. To further characterize the effects of estrogen and antiestrogens, the tissues that respond to estrogen (such as the uterus and the sheath) are immediately removed from the connective and adipose tissue, and the weighed Lu is prepared to pass Image Pr〇epius(Media

Maryland, USA) for image analysis to assess the thickness of the endometrium. In short, the uterus is fixed in 10% fumarin and embedded in stone wax paper. Hematoxylin and eosin stained sections of the mouse uterus were analyzed. Analyze 4 images of each uterus (2 per uterine horn). The average epithelial cell tropism in all animals in each group was measured.

AMJLM

This paper scale applies to the Chinese National Standard (CNS) A4 specification (210 297 mm) A7 1293251 ------_JBT^_ V. Description of invention (SI) Tumor response is assessed at the end of the study, or death of each animal Time (if death occurs in the subject of the experiment) assessment. In such cases, only data from mice that survived at least half of the study (84 曰) were used for tumor response analysis. In short, complete degeneration means that those tumors cannot be measured at the end of the treatment; partial degradation corresponds to those tumors degenerate > 50% of the original volume; stable response refers to tumor regression < 50% or 50% of evolution, and Evolution refers to evolution of more than 50% when compared to the original volume of the tumor. Statistical Analysis Repeated measurements of total tumor surface area change between day 1 and day 161 were analyzed by ANOVA. Patterns include processing, time, and time—the effects of processing interactions plus the conditions of the hierarchy when considering random selection. Therefore, at day 161, meaningful differences between different treatments were tested by time-processing interactions. Residual analysis means that the measurement of the original scale is not suitable for analysis using ANOVA and is not suitable for any attempted conversion. So choose to use the ranking for analysis. This treatment is evaluated for the effect of epithelial thickness using a one-way and also including the hierarchical AN_va at the time of random selection. Inductive pairing comparisons are performed using least squares mean statistics. The error rate (a) for all Type 1 is controlled at 5% to claim a meaningful difference. All singularities were performed using Pr〇c MIXED on SAS Software (SAS Institute, Carry, NC). Results Antagonism of ZR-75-1 tumor growth Treatment of female hormone alone (0VX + E1) resulted in a 707% increase in tumor volume of ZR-75-1 during the 23 weeks of treatment (Fig. 19A). For the female stimulating paper scale, the Chinese National Standard (CNS) A4 specification (210 X 297 mm) is applied ^ ---------^ (Please read the note on the back and fill out this page) Property Bureau Staff Consumer Cooperative Printed 85 A7 1293251 ______B7____ V. Description of Invention (82) (Please read the note on the back and fill out this page) The mouse is administered a daily oral dose of 50 / zg of pure anti-estrogen EM-652 .HC1 completely prevents tumor growth. In fact, not only tumor growth was prevented, but after 23 weeks of treatment, the tumor volume was reduced to 26% of the initial value at the start of the treatment (ρ < 〇 · 〇 4). The value obtained was not statistically different after treatment with EM-652.HC1 and the value observed after oophorectomy alone (OVX), with the tumor volume being less than the initial tumor volume, a 61% reduction. The other six antiestrogens did not reduce the average initial tumor volume during the same dose (50 " g) and treatment. Tumors in these groups were significantly higher than the OVX control group and the group treated with EM-652.HC1 (ρ < 0·01). In fact, compared with the pretreatment values, Droloxifene, Toremifene, GW 5638, Ral〇xifene, Tamoxifene and Idoxifene were treated for 23 weeks. The average tumor volume was higher than the pre-treatment values of 478%, 230%, 227%, 191%, 87%, and 86% (Fig. 19A). -Line - ZR-75-1 tumor growth effect after treatment 161 ,, 200 g per dose of Moxifen Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative (Tamoxifene), no female supplement Next, the average tumor volume was increased to over the reference value of 196% (for OVX, ρ < 〇 · 〇 1) (Fig. 19B). On the other hand, the average tumor volume of these treated Idoxifene mice was increased (125%) (ρ<〇·〇1)' and the tumor volume of these treated dorolifene (T〇rernifene) mice Increase by 86% (ρ < 0·01) (Fig. 19B). Addition of 200 # g EM-652.HC1 to 200 // g Tamoxifene completely inhibited the proliferation observed with Tamsxifene alone (Fig. 19C). On the other hand, at the end of the experiment, the paper scale applies to the Chinese National Standard (CNS) A4 specification (210 X 297 mm). 86 A7 1293251 _____B7 -- -----~-_________ V. Invention Description (83) Separate Treatment of EM-652.HC1 (ρ=〇·44), rai〇xifene (ρ=0·11), Droloxifene (p=〇36) or Gw 5638 (please read the back) ; i want to fill out this page again) (Ρ = 〇 · 17) compared with the OVX control group does not meaningfully change the zR_ 75-1 tumor volume. (Fig. 19B). Effect on range response 〇5 Anti-estrogen effect on female stimulating effect Ministry of Economic Affairs Intellectual Property Office Employees Consumption Cooperative printed in addition to the effect on tumor volume, the response achieved by each individual tumor at the end of the experiment Range is an important parameter for processing efficacy. In ovariectomized mice, complete, partial, and stable tumors reached 21%, 43%, and 38%, respectively, and no tumor evolution. On the other hand, 1% of tumors have been evolved in female-supplemented OVX animals (Fig. 20A). In the feminine supplement 〇νχ animals treated with EM-652.hc1, complete, partial and stable responses were found to be 丨7%, 丨7% and 6〇, respectively. / 〇 tumors and only 7% (2 of 30 tumors) have evolved. In the same situation as female stimulation, treatment of any other antiestrogens at a dose of 5 〇 eg does not reduce the percentage of tumor evolution to less than 6%. In fact, 65 〇/〇 of the tumors (17 of the 26 groups) evolved in the group treated with simamfen (ene), while the treatment of dolomofen (T〇remifene) 89〇/〇 evolution (28 groups) Of the 25 groups), treatment of Rasiixifen (Rai〇xifene) 81〇/〇 evolution (21 of 26 groups), treatment of Droxine (Dr〇i〇xifene) ι〇〇0/〇 evolution (in 23 groups) 23 groups)' treatment of Idoxifen (Idfxifene) η% evolution (20 of the 28 groups) and treatment of Gw 5638 77〇/〇 evolution (2 of the 26 groups) (Fig. 20A). In the case of feminine stimulation, 2〇〇g anti-estrogen is applicable to the Chinese National Standard (CNS) A4 Regulation (210 X 297 public interest) for the paper scale. 87 1293251 A7 Ministry of Economic Affairs Intellectual Property Bureau Staff Consumption Cooperative The effect of the scope of the printed B7___5, invention description (84). As illustrated in Figure 20B, Tamoxifene, Idoxifene, and Toremifene resulted in a higher rate of tumor evolution than other antiestrogens in the absence of feminine stimulation. In fact, after treating 200 ng of each dose of Tamoxifene, Idoxifene and Toremifene, 62% (16 of 26) and 33% (24) The tumor lines of the 8 groups) and 21% (6 of the 28 groups) belong to the evolutionary range. As seen in Figure 20C, when EM-652.HC1 was added to its Tamoxifene (two of the 28 groups), the addition of 200 //g EM-652.HC1 to its Tamoxifene was reduced. The percentage of tumor evolution of Tamoxifene alone was from 62% (16 of the 26 groups) to 7%. Effect of antiestrogens on the thickness of uterine epithelial cells. In the endometrium, the epithelial cells of the endometrium are highly measured as the most direct parameter for the co-effect and antagonism of each compound. The effect of 50 #g antiestrogens per sputum on the thickness of uterine epithelial cells without the stimulation of female hormones. At a daily oral dose of 50 // g, EM-652-HC1 inhibited the epithelial height by 70% after female stimulation. The efficacy of the other six antiestrogens tested was significantly lower (ρ < 0·01). In fact, Droloxifene, GW 5638, Raloxifene, Tamoxifene, Toremifene, and Idoxifene inhibited female stimulating by 17% and 24%, respectively. 26%, 32%, 41% and 50% (Table 10). (Please read the precautions on the back and fill out this page) J.. Order: -Line· 4· This paper scale applies to China National Standard (CNS) A4 specification (210 X 297 mm) 88 1293251 A7 _ B7_ V. Invention Description (85) The effect of 200/g anti-estrogen daily on the thickness of uterine epithelial cells in the absence of female stimulating. In the absence of estrogen stimulation, EM-652.HC1 and Droloxifene were the only two test compounds that did not significantly increase the height of epithelial cells (the values of the OVX control group were 114% and 101, respectively). %) -τ . It has a significant uterine epithelial stimuli relative to the OVX control group, Tamoxifene (155%), Toremifene '135% (135%), and Idoxifene (176%). (Ρ<〇·〇1). Raloxifene (122%) and GW 5638 (121%) also produced a highly statistically significant stimulus for a pair of uterine epithelium (for the OVX control group, p < 0.05 (Table 10)). The same effect and antagonism of each antiestrogens on the measurement of uterine and sheath weight was consistent with that observed for uterine epithelial thickness (data not shown). (Please read the notes on the back and fill out this page)

T Ministry of Economic Affairs Intellectual Property Bureau Staff Consumer Cooperative Printed This paper scale applies China National Standard (CNS) A4 specification (210 X 297 mm) 89 A7 Ministry of Economic Affairs Intellectual Property Bureau Staff Consumer Cooperative Printed Garment 1293251 B7 V. Invention Description ( 86) Endometrial epithelial thickness group η (/^m) soil SEM OVX control group 14. 18.31 soil 004 OVX+E1 control group 8 40·58μ soil 0.63 OVX+Ei +EM-652.HC1 14 25.06b soil 0.07 OVX+E1 + it moxifen 10 33.44b'd soil 0.04 OVX+E, + doramifen 13 31.47 Μ soil 0.04 OVX+E! + la rosifene 12 34.72bd soil 0.06 OVX+E, + dracoxifen 12 36·71μ soil 0.12 OVX+E1 + Idoxifen 12 29.35m soil 0.05 OVX+E! + GW 5638 12 35.30m soil 0.07 OVX + EM-652.HC1 12 20.79 soil 0.10 OVX + it moxifen 11 28.47bd soil 0.05 OVX + EM -652.HC1+ it moxifen 13 27.95bd soil 0.06 OVX + doramifen 13 24.75bc soil 0.04 OVX + lalocifene 12 22.333 soil 0.05 OVX + dracoxifen 13 18.50 soil 0.07 OVX + Idoxifen 11 32.41M soil 0.05 OVX + GW 5638 13 22.223 soil 0.05 ab experimental group relative to OVX control mice: aP&lt ;0.05 ; bP<0.0 1. Cd experiment on EM-652.HC1 treated mice: c Ρ <0·05 ; d P< 0·0 1. (Please read the notes on the back and fill out this page) # Order: JP · This paper scale applies to China National Standard (CNS) A4 specification (210 X 297 mm) 90 Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printed 1293251 λ : ______Β7 _ V. INSTRUCTIONS (87) EXAMPLE 9 Radioactivity Example 9 in the brain of female rats after taking a single oral dose (20 mg/kg) of 14C-EM-800 showed a single oral dose (20 mg) /kg) After 14C-EM-800, the radioactivity in the rat brain. For comparison purposes, blood, plasma, liver, and uterine readings obtained from such animals are also included. " These results are from LREM Research No. 1129: Tissue Distribution and ’ Excretion of Radioactivity Following a Single Oral Dose of

14C-EM-800 (20 mg / 2 ml / kg) to Male and Female Long-Evans Rats. These values indicate that the total amount of drug-derived radioactivity is very low in the brain of female Long-Evans rats (ng equiv/g tissue) and cannot be measured after 12 hr treatment dose. At 2 hours, the brain's radioactivity was 412 lower than the liver, 21 times lower than the uterus, 8.4 times lower than blood, and 13 times lower than plasma. Because the total radioactivity of the brain in unknown proportions is contaminated by blood radioactivity, the value of brain radioactivity shown in Table > 1 is 14C (EM-800) related radiation of a pair of brain tissues themselves. High valuation of active levels. This data shows that the level of anti-estrogens in brain tissue is too low (if present) to counteract the effects of exogenous sex hormones. It is important to note that some of the radioactivity measured in brain tissue may be due to residual blood in the tissue. In addition, the radiochemical purity of 14C-EM-800 used in this study was the lowest 96.25%. This paper scale applies to the Chinese National Standard (CNS) A4 specification (210 X 297 mm) --------------------- Order·-------- (Please read the back of the note first and then fill out this page) 91 1293251 A7 ____ B7 V. Description of invention (88) Table 11 ^隆ί^t^14C-EM.800 (20 mg/kg)i| k choose, and the woven music in the middle of the radioactivity of the semi-robbing,: general, μ Ρ λ / ί_δηη go to the county / ancient knowledge I woven) a: the average concentration (ng EM-800 equivalent / gram group ~ Time (hours) Brain β Blood j Average b (°/〇cv) Average b (%cv) Average b (%cv) 2 17.6 (29) 148.7 (22) 224.6 (20) ~j 4 17.1 ( 29) 66.9 (45) 103.2 (39) 6 15.6 (8) 48.3 (29) 74.1 (31) 8 _ 16.8 (31) 41.1 (12) 64.1 (14) 12 10.0C (87) - 28.7 (54) 40.7 ( 55) 24 0 (NC) 4.7d (173) 10.1 (86) 36 0 (NC) 0 (NC) 0 (NC) 48 0 (NC) 0 (NC) 0 (NC) 72 0 (NC) 0 (NC 0 (NC) 96 0 (NC) 0 (NC) 0 (NC) 168 0 (NC) 0 (NC) 0 (NC) a: LREM 1129 from the report (EM-800: for single oral administration) Dosage: ^14C-EM-800 (please read the precautions on the back and fill out this page). and the radiation of excrement b: 1.2 ng EM-800 equivalent limit of quantitation (LOQ) c : one sample below LOQ; 0 d for calculating the average: two samples below LOQ; 0 % CV for calculating the average : coefficient of variation, expressed as a percentage, where n = 3. ||nc: not calculated. -ΜΜΨ . Ministry of Economic Affairs, Intellectual Property Office, Staff Consumer Cooperative, printed paper scale applicable to China National Standard (CNS) A4 specification (210 X 297 PCT) 92 1293251 V. Description of invention (89)

Printed by the Intellectual Property Office of the Ministry of Economic Affairs, Employees' Consumer Cooperatives a : LREM丨129 The value obtained from the report (EM-800: tissue distribution and radioactivity of excreta in rats given a single oral dose). b : 1.2 ng EM-800 equivalent limit of quantitation (LOQ). c : one sample below LOQ; 0 is used when calculating the average. d : 2 samples below LOQ; 0 is used to calculate the average. %CV: coefficient of variation, expressed as a percentage, where η = 3. NC : Not calculated. 〇\ 00 Os to Lk) ON to to 00 ON to Time (hours) Average concentration of female Long-Evans rats after treatment with a single oral dose of l4C-EM-800 (20 mg/kg) /EM-800 eq/g tissue)a 〇ooooo 0.0100 0.0168 0.0156 0.0171 0.0176 Card ί η < Brain (NC) (NC) (NC) (NC) (NC) (NC) ^00 ^00 "to 0.0348 0.0944 0.1326 0.2196 0.4034 0.6391 1.8232 2.7748 2.7462 3.2201 7.2547 Card Vv- i η < Liver Η—^ § ^00 ^00 % η ΓΠ 2 1 〇〇〇ο 彐σο^ to 彐r Λ 爻r 3 CTP 3 匕> 〇o O 0.0238 0.0261 0.0837 0.2407 0.3332 0.2757 0.2866 0.3675 Average b (%cv) Uterus (NC) (NC) (NC) N •—A 13 s Lk) § ooo 〇o 0.0047d 0.0287 0.0411 0.0483 0.0669 0.1487 η < Blood, Selecting Drug-Derived Radioactivity in Tissue (NC) (NC) (NC) (NC) (NC) (173) OS ooooo 0.0101 0.0407 0.0641 0.0741 0.1032 0.2246 V4- (Τ 7 η < Plasma (NC) (NC ) (NC) (NC) (NC) 00 N Lk) § s ------------- loading .------- order ·------- line ( Please read the notes on the back and fill in Page) This paper scale applies to China National Standard (CNS) A4 specification (210 X 297 mm) 93 A7 1293251 _ B7_ V. Invention description (9〇) Example 10 (Please read the note on the back and fill in this page) Combination of estrogen EM-652.HC1 and estradiol protects against uterine irritant materials and methods Animals and treatments used in female Spock-Dawley rats (10- to 12 weeks old) and weighing 215-265 g at the time of ovariectomy (CrLCD (SD) Br) (Charles River Laboratory, St-Constant, Canada). The animals were individually placed in an environmentally controlled room (temperature: 22 ± 3 °C; humidity: 50 ± 20%; 12 hours of 12-hour dark cycle, starting at 07:15). These animals are allowed to drink free tap water and approved dental caries (Lab Diet 5002 (granules), Ralston Purina, St-Louis, M0). The experiments were carried out in animal equipment approved by the Canadian Council on Animal Care (CCAC) and the Association for Assessment and Accreditation of Laboratory Animal Care (AAALAC) in accordance with the CCAC Guide for Care and Use of Experimantal Animals. ^ 137 rats were randomly assigned to 10 groups of 13 or 14 animals in each group of the Ministry of Economic Affairs, the Intellectual Property Office staff consumption cooperative printing group, such as: 1) complete control group; 2) ovariectomy (OVX) control Group; 3) OVX + 17/5-estradiol (E2; 2 mg/kg); Groups 4 to 10) OVX + E2+ EM-652.HC1 (0.01, 0.03, 0.1, 0.3, 1, 3 or 10) Mg/kg). In the first study of this study, a suitable group of animals (OVX) underwent bilateral ovaries under anesthesia with Isoflurane. From the first to the 14th day of the study, the test compound (0.5 ml/rat) suspended in 0.4% methylcellulose was administered once a day by oral intensive feeding. Groups 1 and 2 of the animal group received vehicle alone only during the same time period. For the 15th paper scale of this study, the Chinese National Standard (CNS) A4 specification (210 X 297 mm) is applied. 94 Ministry of Economic Affairs Intellectual Property Office Staff Consumer Cooperative Printed 1293251 λ: __Β7_ V. Invention Description (91) 曰, each Four animals in the group were injected with 10% buffered formalin and the tissues were examined for tissue. Under the anesthesia of Isoflurane, other animals are killed by bloodletting at the aorta. The uterus and sheath are removed, the remaining fat is removed and weighed. Samples from each uterus were fixed with 10% buffered formalin and a computer-assisted program (Software Image-Pro Plus) was used to determine the height of the endometrial epithelial cells. Serum Cholesterol Level The Boehringer Mannheim Diagnostic Hitachi 911 Analyzer (Boehringer Mannheim Diagnostic Laboratory Systems) was used to measure the total cholesterol content in jk clear samples collected from overnight unfed animals. Statistical analysis Data are expressed as mean soil SEM. Statistical significance was determined according to the Duncan-Kramer (Kramer, Biometrics, 12: 307-310, 1956) multi-range test. Results t The 65% reduction in uterine weight observed 2 weeks after ovariectomy was completely recovered by daily oral administration of 2 mg/kg of 17泠-estradiol (E2) (490±26 mg vs. 480±) 17 mg ; NS·) (Fig. 21). As exemplified in the same figure, the gradual inhibition of E2-stimulated uterine weight was observed when the EM-652.HCM dose was increased, and 84% and 87 were observed in the anti-estrogen doses of 3 mg/kg and 10 mg/kg, respectively. % of E2 responds. Similar results can be obtained from the epithelial height of the endometrium (Fig. 22). In fact, the dose of 3 mg / kg paper size applies to China National Standard (CNS) A4 specifications (210 X 297 mm) -------------Package-------- -------- Line (please read the notes on the back and fill in this page) 95 Ministry of Economic Affairs Intellectual Property Bureau Staff Consumer Cooperative Printed 1293251 Λ7 ---- B7___ V. Invention Description (92) and 10 mg /kg of antiestrogens prevented 83% and 93% of sputum from stimulating endometrial epithelial height, respectively. Compared with the intact control animals (31.1 ± 0.7 mm), the epithelial density of the endometrium of the 〇νχ animal group (41 9 soil 1.2 mm) treated with the E2 compound was higher (34.7%, p< 〇.〇1) (Fig. 23). The sheath weight of E2 supplemented OVX animals was similar to that of intact animals (149.9 ± 9.0 mg vs. 145 ± 5.3 mg, N.S.). As seen in Fig. 24, inhibition of sheath weight was observed using a higher dose of ΕΜ-652·Ηα, as compared to uterine weight. In fact, no significant anti-estrogen inhibition of the weight of the sheath was observed until the treatment of this compound at i mg/kg. In fact, the 3 mg/kg dose of EM-652.HC1 resulted in a 50% inhibition of E2 stimulation by a single stone. The effect of E2 on the weight of the sheath was observed when the dose of antiestrogens was treated. Complete reply. A 37% increase in serum cholesterol was observed 2 weeks after OVX (p<〇 〇 1). On the other hand, treatment of OVX animals with E2 resulted in a 53% 0<〇〇1) serum cholesterol level inhibition (Figure 25). There was no statistically significant effect on the inhibition of E2 by the addition of EM-652.HC1 at a dose of mg1 mg/kg to 〇3 mg/kg per dose. On the other hand, EM_ 652.HC at doses of 1.0, 3.0 and 1 〇 mg/kg reduced the sputum effects by 36%, 30% and 50%, respectively. This data clearly demonstrates that anti-estrogen, EM-652 HC1 neutralizes the stimulatory effect of h on the epithelial height of the eucommia and endometrium, and the uterine weight and epithelial height of the lining of the lining are two confirmed peripherals. The estrogen action parameters of the tissue. The data clearly show that in women who receive estradiol to relieve vasomotor symptoms after menopause, a total of EM_652 HC1 (please read the back first; 1) and then fill out this page - Λ · -- line · -xf - This paper scale applies to China National Standard (CNS) a4 specification (210 X 297, hair) 96 1293251 A7 B7 V. Invention description (93 administration will avoid estrogen stimulation of the endometrium EM-652.HC1 inhibition at doses of 1.0 mg/kg, 3.0 mg/kg, and 10 mg/kg recovered by 36%, 30%, and 50% may be explained by the advantage of antiestrogenic effects, where Estrogen action may result in the same degree of inhibition as when used alone. In fact, SEM-652.HC1 has a 5-fold higher affinity for rat uterus ER than E2 itself (Martel et al., J. Steroid Biochem. molec. Biol) 64: 199-205, 1998. Examples of Pharmaceutical Compositions and Kits The following description (illustrated by way of example and not limited to this method) is based on several preferred activated SERMs EM-800 or EM-652. HC1 (EM1538) and preferred activated estrogen 17/5-estradiol, ethinyl estradiol Pharmaceutical compositions and kits of conjugated estrogen. Other compounds of the invention or combinations thereof may be used in place of (or in addition to) EM-800 or EM-652.HC1 or 17 /5-estradiol or ethynyl erectin Alcohol. The concentration of the activating starting material can generally vary by a wide range as discussed herein. The amounts and types of other starting materials that may be encompassed by this study are well known in the art. ------^--------^---------^ (Please read the notes on the back and fill out this page) t' Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative The paper size is applicable to the Chinese National Standard (CNS) A4 specification (210 X 297 mm). 97 1293251 A: _B7 V. Inventive Note (94) Example A Pharmaceutical composition for oral administration (capsule) Raw material% by weight Total composition weight) EM-652.HC1 5.0 Ethynylestradiol 0.02 hydrated lactose 79.98 Starch 4.8 Cellulose crystallites 9.8 Magnesium stearate 0.4 or raw material weight% (% of total composition) EM-652.HC1 5.0 Estrogen 0.2 hydrated lactose 79.98 Starch 4.8 Cellulose crystallites 9.8 Magnesium stearate 0.4 (Please read the back first Note: Please fill out this page) - Λ - - - Ministry of Economic Affairs Intellectual Property Bureau Employees Consumption Cooperatives Print Sample B Set of SERM and steroids are administered by oral administration of non-steroidal antiestrogens for oral administration ( Capsules) % by weight of raw materials (by weight of total composition) EM-652.HC1 5.0 Hydrated lactose 80.0 Starch 4.8 Cellulose crystallites 9.8 Magnesium stearate 0.4 This paper scale applies to China National Standard (CNS) A4 specification (210 X 297 PCT) -- Line - 98

Printed by the Intellectual Property Office of the Ministry of Economic Affairs, Department of Consumers' Cooperatives, Printing 1,293,251 V. Description of the Invention (95 % by weight of raw materials - (by weight of total composition) Ethyl estrone 0.02 hydrated lactose' " ~ ---- 84.98 4.8 Cellulose microcrystals ~~ ~~'~ 9.8 Stearic acid town ~~ ~ - 0.4 In the above formula, other SERMs can be replaced to replace Μ·800 or EM 01538, and other estrogens can be replaced instead of 丨7 _ estradiol, ethinyl estradiol or conjugated estrogen. In the following cases, more than one type of SERM or more than one type of estrogen may be included, and the combined weight percentage is preferably a single estrogen given in the above examples. Or the weight of a single-SERM, the invention has been described for the preferred embodiments and examples, but is not limited to the specific examples and examples. Those skilled in the art should be able to easily recognize the broader The scope of the invention is limited only by the scope of the invention as described herein. The k-paper scale applies to the Chinese National Standard (CNS) A4 specification (210 X 297 public f 99)

Claims (1)

The scope of application for patents is like a mixture. 4. A pharmaceutical composition comprising: a) a pharmaceutically acceptable adjuvant, diluent or carrier; b) a therapeutically effective amount of at least - an estrogen or a prodrug thereof; and C) - at least a therapeutically effective amount of a selective estrogen receptor modulator or a compound wherein the selective estrogen receptor modulator is a compound different from the estrogen; the selective estrogen receptor modulator is not the following Compounding m-derivatives, -naphthalene derivatives, _isophorin derivatives or a 3 stupid 4-base porphyrin derivative having a configuration of more than _ 2R mirrors - 3-phenyl thiochroman (chr〇man). A mirror image mixture of organisms and 3-phenylchroman derivatives. 5. A kit comprising a first container comprising a therapeutically effective amount of a pharmaceutical formulation of at least one estrogen or a prodrug thereof, and the kit further comprising a second container, The second container comprises a pharmaceutical formulation of a therapeutically effective = at least one selective estrogen receptor modulator or a prodrug thereof, the selective estrogen receptor modulator being not a benzothiphenol derivative. 6. A pharmaceutical composition comprising: a) a pedigree acceptable supplement, diluent or carrier; b) - a therapeutically effective amount of at least one estrogen or a prodrug thereof; c) at least one A therapeutically effective amount of a selective estrogen receptor modulator or a prodrug thereof, wherein the selective heterohormone receptor modulator is a compound different from estrogen; and d) - a therapeutically effective amount of at least one additional agent The reagents are selected and patented from the group consisting of bisphosphonate, androgen reagent, citridin, dehydroepiandrosterone, dehydroepiandrosterone sulfate, and male fat. Alkene 3/5, 17 A-diol, 4· androstene-3,17-dione, and any of the aforementioned prodrugs. 7. The pharmaceutical composition of claim 2, wherein the pharmaceutical composition further comprises a therapeutically effective amount of a bisphosphonate. a pharmaceutical composition for reducing or eliminating the incidence of menopausal symptoms, the pharmaceutical composition comprising a therapeutically effective amount of estrogen or a prodrug thereof in combination with a therapeutically effective amount of a selective estrogen receptor modulator or The prodrug, the selected estrogen receptor modulator is a compound different from estrogen, the pharmaceutical composition further comprising a therapeutically effective amount of at least one additional reagent selected from the group consisting of Group consisting of: dehydroepiandrosterone, dehydroepiandrosterone sulfate, androgen reagent, 睪崔睪同, androgen-5-ene-3^17/3-diol, 'androsenene _ 3,17-dione and any of the aforementioned pre-addition reagents. 9. A pharmaceutical composition for treating or reducing the risk of developing a condition selected from the group consisting of osteoporosis, hypercholesterolemia, hyperlipidemia, atherosclerosis, hypertension , Azheimer's disease, insulin resistance, diabetes, dysfunction, obesity, vaginal bleeding caused by hormone replacement therapy, and breast tenderness caused by hormonal therapy, the pharmaceutical composition comprising a therapeutically effective amount of estrogen or The prodrug is combined with a therapeutically effective amount of a selective estrogen receptor modulator or a prodrug thereof, the selective estrogen receptor modulator being a compound different from estrogen, further comprising at least one therapeutically effective amount Add test 1293251 ", the patent application range agent, the added reagent is selected from the group consisting of: dehydroepiandrosterone, dehydroepiandrosterone sulfate, male fat _5_ ene, cold Monool, androgen reagent, trifluralone, 'androstene-3,17-dioxin and any of the foregoing prodrugs. The kit comprises a first container comprising a therapeutically effective amount of a pharmaceutical formulation of at least one estrogen or a prodrug thereof, and the kit further comprises a second container The second container comprises a therapeutically effective amount of a pharmaceutical formulation of at least one selective estrogen receptor modulator or a prodrug thereof, the kit comprising at least one additional container comprising at least one selected from the group consisting of Therapeutically effective amounts of the following groups of added agents··Dehydroepiandrosterone, dehydroepiandrosterone-sulfate, male lipid _ ene-3 stone, diol, androgen reagent, ketone, 4 • Androstene 3, 17-dione and any of the aforementioned prodrugs. U. The kit of claim 5, which is useful for treating or reducing the risk of suffering from osteoporosis, the kit comprising at least one additional container comprising a therapeutically effective treatment of at least one bisphosphonate A quantitative pharmaceutical formula. Generic application for patent scope! The pharmaceutical composition of the formula further comprises a therapeutically effective amount of the androgen reagent. The pharmaceutical composition of claim 1, wherein the selective estrogen receptor modulator has a molecular formula having the following characteristics: a) two aromatic rings separated by 1 to 2 intermediate carbon atoms, The two aromatic rings may be substituted or unsubstituted by a group or a group converted into a group in a living body; 103' 1293251 t, patent application b) - one having an aromatic ring and a tertiary amine a side chain of a functional or a salt thereof; and wherein the selective estrogen receptor modulator is not the following compound: a benzotriazine derivative, a naphthalene derivative, or a different one. Mirror image of quinolin derivative or one of 3 phenylquinoline derivatives, 3-phenyl thiochroman derivatives and 3-phenyl chroman derivatives with more than 10% 2R mirror configuration mixture. 14. The pharmaceutical composition of claim 13, wherein the side chain is selected from the group consisting of the following chemical formulas: 15. The pharmaceutical composition of claim 13, wherein the selective estrogen receptor modulator is selected from the group consisting of triphenylethylene derivatives, σ derivatives, benzene ° ratio σ South derivative, HMR 3 3 3 9 , HMR 3656, LY 335124, LY 326315, SH 646, ERA 923 and centchroman derivatives. 16. The pharmaceutical composition of claim 8, wherein the selective estrogen receptor modulator is a benzothiophene derivative of the following formula 104 1293251 Wherein IMaR2 is independently selected from the group consisting of hydrogen, hydroxyl and a moiety converted into a hydroxyl group in a living body; wherein R3 and R4 are independently alkyl groups of from 丨 to ^# or ^) A portion, which is combined with the nitrogen to which they are bound, is selected from the group consisting of: σ ratio bite, dimethyl chlorpyrifos, fluorenyl-tetradecyl An imido group, a hexahydropyridine, a hexamethylene imine, and a linne; wherein the lanthanide is selected from the group consisting of _c〇_, -CtI〇H, and _Ch2-; wherein the B is selected from A group consisting of a phenylene group, an acridinium group, and a _ ring C4h2N2-. 17. The pharmaceutical composition of claim 16, wherein the selective estrogen receptor modulator is selected from the group consisting of Ral〇xifene, LY 353381, and LY 335563. 18. The pharmaceutical composition according to claim 13, wherein the selective estrogen receptor modulator is a triphenylethylene or diphenylhydronaphthalene derivative compound having the following chemical formula: 105 1293251 ^, claiming a patent range Wherein D is -0 (: Η2 (3Η2Ν(Π3)Ϊ14, -OCH2CH2OH or -CH=CH-COOH (R3 and R4, which may be independently selected from the group consisting of C!-C4 alkyl) Or R3, R4 and the nitrogen to which they are bonded together constitute a ring structure selected from the group consisting of pyrrolidine, dimethyl-1-pyrrolidine, methyl-1 -tetramethylimine Base, hexahydropyrene ratio bite, hexamethylene imine and morphine); wherein ugly and lanthanide are independent hydrogen or hydroxy, phosphate or lower alkyl, wherein J is hydrogen or halogen. 19. The pharmaceutical composition of claim 1, wherein the selective estrogen receptor modulator is selected from the group consisting of: 0Η-Tamoxifene, Droloxifene, and more Toremifene, Iodoxifene, Lasofoxifene, iproxifene, FC1271 and GW5638. 20. The pharmaceutical composition according to claim 13, wherein the selective estrogen receptor modulator is one liter of the quinone derivative compound of the following formula: 106 1293251, the scope of patent application Ri- rrVcr wherein D is selected from -〇CH2CH2N(R7)R8, -CH=CH-CO N(R7)R0. a group of -CC-(CH2)nN(R7)R8 (R7 and 8 can be independently selected from the group consisting of CrC6 alkyl groups, or R7, R8 and their linkages The nitrogen together constitutes a ring structure selected from the group consisting of ϋ 嘻 bite, 曱 曱 。 比 比 比 比 比 比 比 比 比 比 比 比 比 比 比 比 比 比 比 比 比 曱 曱 曱 曱 曱 曱 曱 曱 曱 曱 曱 曱 曱And X Lin) wherein X is selected from the group consisting of hydrogen and CrC6 alkyl; wherein R!, R2, R3, R4, 5 and R6 are independently selected from hydrogen, hydroxy, CKC6 alkyl And a group consisting of a portion converted into a hydroxyl group in a living body. 21. The pharmaceutical composition according to claim 20, wherein the selective estrogen receptor modulator is TSE 424 (2-(4-hydroxyphenyl)methyl small [[4-[2-(1) - hexahydroacridinyl) ethoxy]phenyl]methyl hydrazine _5_ alcohol). 1293251 VI. Patent application scope (centchr〇man) derivative compound·· D Wherein Ri and R2 are independently selected from the group consisting of hydrogen, a hydroxyl group and a moiety converted into a mesogenic group in a living body; wherein I and R0 are independently hydrogen or a Cl-c6 alkyl group, wherein the D system is and R4 The ring structure may be independently selected from the group consisting of Ci_C4 alkyl groups, or the ruler 3, hydrazine and the nitrogen to which they are bonded together constitute a ring structure selected from the group consisting of pyrrolidine and dimethyl small. Pyridoxine, methyl-1-tetramethylimido, hexahydropyrene, hexamethyleneimine and morphine). And the pharmaceutical composition according to claim 22, wherein the centchroman derivative is (3,4_trans-2 2-didecylphenyl_4_[4-(2-(2) - (tetramethylimine-1-yl)ethoxy)phenyl)-7-methoxy 2 roman. 24. The pharmaceutical composition of claim 13, wherein the selective gonadotropin receptor The regulator has the following molecular formula·· --------. 108 1293251 The center and the ruler 2 are independently hydrogen, hydroxyl or a part converted into a hydroxyl group in a living body; wherein the Z system is a vacancy or is selected from -CH2-, ·0-, -S-, and -NR3-(I a group consisting of hydrogen or lower alkyl; wherein the R is a divalent moiety, the divalent moiety is from about 4 to 10 intermediate atoms from the ring; wherein L is a divalent moiety Or a trivalent moiety, the divalent or trivalent moiety is selected from the group consisting of -SO·, {〇Ν., _Ν< and other N<; wherein 〇 is selected from hydrogen, (^ a group consisting of a hydrocarbon, a combination of a divalent moiety, and [a 5- to 7-membered heterocyclic ring, and the aforementioned dentate or non-derivative derivative; wherein the G2 is vacant or selected from a group consisting of hydrogen, (^ to ^ hydrocarbon, a combination of a G and a divalent moiety, and [a 5- to 7-membered heterocyclic ring, and the aforementioned halogenated or unsaturated derivative; wherein & a pharmaceutical composition selected from the group consisting of hydrogen, a methyl group, and an ethyl group, 129, 129, 251, and the pharmaceutical composition of claim 24, wherein the compound is Phenyl pyran derivative having the following general structures: Or a pharmaceutically acceptable salt thereof, wherein D is -OCH2CH2N(R3)R4 (R3 and R4 may be independently selected from the group consisting of CKC4 alkyl groups, or R3, R4 and their linkages The nitrogen together constitutes a ring structure selected from the group consisting of pyrrolidine, dimethylpyrrolidine, methyl-1-tetramethylimido, hexahydroindole, hexamethyleneimine, and Wherein Rjn R2 is independently selected from the group consisting of hydrogen, a hydroxyl group, and a moiety which is converted to a hydroxyl group in a living body. 26. The pharmaceutical composition of claim 25, wherein the benzofuran derivative is optically active, since most of its stereoisomers have an S absolute configuration at the second carbon, the compound having the following Molecular structure: 110 1293251 t, patent application scope 5· Wherein Ri and R2 are independently selected from the group consisting of a hydroxyl group and a moiety which can be converted into a hydroxyl group in a living body; wherein R3 is a species selected from the group consisting of saturated, unsaturated or substituted four Hypomethylimido, saturated, unsaturated or substituted hexahydrohydrogenated 'saturated, unsaturated or substituted hexahydropurine π group, saturated, unsaturated or substituted morpholine, nitrogen-containing cyclic moiety, nitrogen-containing Polycyclic moiety and NRaRb (Ra and Rb are independently hydrogen, linear or branched iCKC6 alkyl 'linear or branched C2-C6 alkenyl and straight or branched C2-C6 alkynyl) 〇27· The pharmaceutical composition of claim 26, wherein the compound or salt is substantially deficient in (2R)-mirror. 28. The pharmaceutical composition of claim 26, wherein the selective estrogen receptor modulator is selected from the group consisting of: 111 1293251 t, patent application scope EM-800 EM- 1520 EM-1872 EM-1900 EM-1901 EM-1903 112 with 1293251, the patent application scope EM-1533 EM-1518 EM-652.HC1 (EM-1538) wherein all of the aforementioned molecular structures, the stereochemistry of which is indicated as optical activation, since most of its stereoisomers are The pharmaceutical composition of claim 26, wherein the benzopyran derivative is a monoacid salt selected from the group consisting of acetic acid and adipic acid. , benzenesulfonic acid, benzoic acid, camphorsulfonic acid, citric acid, trans-butenedioic acid, hydroiodic acid, hydrobromic acid, hydrochloric acid, thiohydrochloric acid, hydroxy-naphthoic acid, lactic acid, cis-butyl Aenedioic acid, methanesulfonic acid, mercaptosulfuric acid, 1,5-naphthalene disulfonic acid, nitric acid, palmitic acid, piavaic acid (pivalic) 113 1293251 6. Patent application range: acid, phosphoric acid, propionic acid, succinic acid, sulfuric acid, tartaric acid, terephthalic acid, p-toluenesulfonic acid and valeric acid. 30. The pharmaceutical composition of claim 29, wherein the acid is chlorous acid. 3^ If applying for a specific composition, the selectivity of the hormone receptor modulator is: EM-652.HC1 (EM-1538) and is optically active because most of its stereoisomers are 2 S configurations; and wherein the estrogen is selected from the group consisting of: 17 y?-estradiol, 17-point estradiol ester, 17α-estradiol, 17"-estradiol ester, female sterol, female sterol ester, female, female conjugate, estrogen , equilin, isoflavone vinegar (equilin, 17 α -ethynyl estradiol, 17 α • ethinyl estradiol ester, mestranol and mestreol) (mestran〇i ester). The pharmaceutical composition of claim 1, wherein the estrogen is selected from the group consisting of 17yS-estradiol, 17 stone _estradiol 酉曰, female sterol, female sterol ester, female, female ester, co-roll estrogen, equilin, eucalyptus vinegar (such as eqUilin), 17 α-ethynyl estradiol, 17α - Ethyl estradiol ester, Mestre alcohol 1293251 6. Patent scope (tran〇l), mestranol ester, chemical estrogen, DES, phenantine Hormone, tibolcme, ethyl estrogen oxazapine and ethane diol. The pharmaceutical composition of claim 1, wherein the selective estrogen receptor is in the breast or uterus There is no estrogenic activity in the endometrial tissue. The pharmacy composition of the patent scope of the patent application, wherein the estrogen is a compound of estrogen/androgen. 5. The pharmaceutical composition of claim 34, wherein the mixed beach 36 hormone/androgen compound is tibolone. ^ : The pharmacy composition of the term i and the range i, in which the menopausal symptoms are composed of the vasomotor symptoms of the tidal vasomotor, irregular menstruation, dryness of the sinus, headache, and sleep disturbance. Group. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition reduces the risk of suffering from breast or endometrial cancer in the patient. 115