Gonadotropin-releasing hormone receptor antagonists and uses thereof

Abstract

The present invention provides a compound or a pharmaceutically acceptable salt, isomer, prodrug, polymorph, or solvate thereof, and a method for preparing and using the compound, which, as a GnRHR antagonist, has activity equivalent to or superior to that of elagolix in calcium flow assays and has better pharmacokinetic properties. [Formula 1] JPEG2023523097000014.jpg5374

Description

[Technical field]

[0001] The present invention relates to the field of organic chemistry, and in particular to a compound or a pharma-ceutically acceptable salt, isomer, prodrug, crystalline polymorph or solvate thereof, and its preparation method and use. [Background technology]

[0002] Gonadotropin-releasing hormone (GnRH), also called luteinizing hormone-releasing hormone (LHRH), is a decapeptide secreted from the hypothalamus. GnRH exerts its biological action by binding to the GnRH receptor located in the anterior pituitary gland, and can stimulate the biosynthesis and secretion of luteinizing hormone (LH) and follicle-stimulating hormone (FSH). These act in concert to stimulate the development of germ cells in the ovaries or testes and the production and secretion of sex hormones.

[0003] Therapeutic agents targeting the GnRH receptor can inhibit hypothalamic-pituitary-gonadal axis function and are widely used to treat gonadal hormone-dependent diseases such as prostate cancer, breast cancer, endometriosis, uterine fibroids, prostatic hyperplasia, assisted reproductive therapy, and precocious puberty.

[0004] Currently used therapeutic agents include GnRH receptor agonists or antagonists. GnRH receptor agonists have been approved for use in the treatment of reproductive hormone-dependent diseases, such as prostate cancer, premenopausal breast cancer, endometriosis, and uterine fibrosis. In the early stages of treatment, GnRH receptor agonists can stimulate pituitary gonadotropic cells to increase the secretion of gonadotropic hormones. This causes a "flare up" effect that lasts for about 1 to 3 weeks. However, long-term administration can cause the gonadotropic hormones to be depleted and their receptors to be downregulated. Thus, after a certain period of time, the gonadal hormones are suppressed, and the circulation of gonadal hormones, such as estrogen, progesterone, and testosterone, is downregulated, resulting in the therapeutic effect of hormone-dependent diseases.

[0005] Contrary to GnRH receptor agonists, GnRH receptor antagonists competitively bind to the GnRH receptors in the pituitary gland, directly and quickly inhibiting the pituitary-gonadal axis, thereby suppressing the peak of endogenous luteinizing hormone and lowering estrogen levels. In this case, the effect is immediate, there is no activating effect, and no "flare-up" effect occurs, so the function of the pituitary gland can be restored 2 to 4 days after the administration is stopped. In other words, it is possible to avoid side effects caused by the "flare-up" effect in the early stage of GnRH receptor agonist treatment. In addition, it is more economical and convenient because it shortens the period of drug use in clinical practice.

[0006] At present, some peptide GnRH receptor antagonists have been approved for sale and are used for assisted reproduction in infertility and for the treatment of late-stage prostate cancer. However, these peptide GnRH analogues cannot be administered orally, and require subcutaneous injection, nasal spray, or long-term sustained-release injection. Therefore, there are drawbacks such as the injection site stimulating histamine release reaction and slow removal after administration is stopped, limiting their application. In contrast, non-peptide GnRH receptor antagonists can be administered orally, and can avoid the pain caused by injection and the allergic reaction caused by histamine release, improving patient compliance and providing further advantages (Non-Patent Document 1).

[0007] Elagolix is ​​the first oral non-peptide GnRH receptor antagonist to be marketed, and is used to treat endometriosis, and is also being studied for use in other estrogen-dependent diseases such as uterine fibroids. Elagolix belongs to the pyrimidine-2,4-dione compound (Guo Zhiqiang et al., Patent Document 1), and competitively binds to the GnRH receptor in the pituitary gland to block the GnRH signal pathway and reversibly reduce the secretion of ovarian hormones, estradiol, and progesterone.

[0008] Elagolix can be orally absorbed, but has only moderate to low levels of cell permeability. In addition, its oral bioavailability is very low, with absolute oral bioavailability in mice, rats, and monkeys being only 10%, 5.8%, and 11%, respectively (Non-Patent Document 2). However, when radiolabeled elagolix was orally administered to rats, nearly 50% of the total radioactivity was excreted in the bile. This suggests that elagolix has a hepatic first-pass effect, and most of the drug may not be able to circulate and exert its effect. As a result, the amount of therapeutic agent used in clinical trials has increased (the recommended clinical dose of elagolix is ​​150 mg, qd, or 200 mg, bid). In addition, the total radioactivity after oral administration to rats was distributed mainly in the digestive tract and liver. Elagolix also showed a large apparent distribution volume (1674 L) in the human body. All of these indicate high tissue distribution characteristics. If distribution to the liver is too large, there may be a risk of hepatotoxicity due to elagolix (increased hepatic aminotransferases) or effects on hepatic drug-metabolizing enzymes (elagolix is ​​a substrate for CYP3A, P-gp, and OATP1B1, and is also an inhibitor of P-gp, and has a weak to moderate inductive effect on P450 (CYP) 3A) (Non-Patent Document 3).

[0009] [ka]

[0010] Despite significant research in the field, there remains a need for effective small molecule GnRH receptor antagonists with more ideal therapeutic effects, as well as pharmaceutical compositions containing the GnRH receptor antagonists and methods of using the same to treat sex hormone-related disease states. [Prior art documents] [Patent documents]

[0011] [Patent Document 1] Chinese Patent Application No. 200480019502.3 [Non-patent literature]

[0012] [Non-Patent Document 1] Sarma,PKS et al,Expert Opinion on Therapeutic Patents 16(6):733-751,2006 [Non-Patent Document 2] Chen Chen,et al.Discovery of Sodium R-(+)-4-{2-[5-(2-Fluoro-3-methoxyphenyl)-3-(2-fluoro-6-[trifluoromethyl]-benzyl)-4-methyl-2,6-dioxo-3,6-dihydro-2H-pyrimidin-1-yl]-1-phenylethylamino}butyrate(Elagolix), a Potent and Orally Available Nonpeptide Antagonist of the Human Gonadotropin-Releasing Hormone Receptor,J.Med.Chem.2008,51,7478-7485 [Non-Patent Document 3] FDA,Orilissa(elagolix),210450Orig1s000MultiD,2018 Summary of the Invention [Problem to be solved by the invention]

[0013] In view of the above-mentioned shortcomings of the prior art, the object of the present invention is to provide a compound or its pharma- ceutically acceptable salt, isomer, prodrug, crystalline polymorph or solvate, and its preparation method and use, in order to solve the problems of the prior art. [Means for solving the problem]

[0014] To achieve the above and other related objects, the present invention provides, in one aspect, a compound or a pharma- ceutically acceptable salt, isomer, prodrug, polymorph, or solvate thereof, the chemical structure of which is shown in Formula I:

[0015] [ka]

[0016] In some embodiments of the invention, the isomers are selected from enantiomers, diastereomers, cis-trans isomers or stereoisomers.

[0017] In another aspect, the present invention provides a method for preparing the above compounds, which comprises hydrolyzing a compound of formula 1-12 to prepare a compound of formula I. The reaction scheme is as follows:

[0018] [ka]

[0019] In some embodiments of the present invention, the hydrolysis reaction can usually be carried out in the presence of a base.

[0020] In another aspect, the present invention provides the use of the above compound or a pharma- ceutically acceptable salt, isomer, prodrug, crystal polymorph or solvate thereof in the manufacture of a medicament.

[0021] In some embodiments of the invention, the drug is selected from gonadotropin releasing hormone receptor antagonists.

[0022] In some embodiments of the invention, the drug is selected from drugs for treating diseases related to gonadal hormones.

[0023] In some embodiments of the present invention, the disease associated with gonadal hormones is selected from endometriosis, menopause, menstrual irregularities, uterine fibroids, uterine fibroids, polycystic ovary syndrome, endometriosis, uterine leiomyoma, lupus erythematosus, hirsutism, precocious puberty, dwarfism, acne, alopecia, gonadal hormone-dependent tumors, pituitary adenomas producing gonadotropic hormones, sleep apnea syndrome, irritable bowel syndrome, premenstrual syndrome, prostatic hyperplasia, contraception and infertility, and Alzheimer's disease.

[0024] In some embodiments of the invention, the gonadal hormone-dependent tumor is selected from prostate cancer, uterine cancer, breast cancer, ovarian cancer, and pituitary gonadotroph adenoma.

[0025] In another aspect, the present invention provides a pharmaceutical composition comprising the above compound or a pharma- ceutically acceptable salt, isomer, prodrug, crystal polymorph, or solvate thereof. [Brief description of the drawings]

[0026] [Figure 1] FIG. 1 shows the drug blood concentration-time curve when G201 / elagolix was intravenously injected into SD rats in Example 3 of the present invention. [Diagram 2] FIG. 1 shows the drug blood concentration-time curve when G201 / elagolix was orally administered to SD rats in Example 3 of the present invention. [Diagram 3] FIG. 13 is a graph showing the drug blood concentration-time curves when G201 was intravenously and orally administered to ICR mice in Example 4 of the present invention. [Figure 4] FIG. 13 shows the trend of changes in serum LH after oral administration of G201 / elagolix to castrated cynomolgus monkeys in Example 6 of the present invention (LH is a percentage value compared to before administration, N=3). DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0027] In order to make the objectives, technical solutions and beneficial technical effects of the present invention clearer, the present invention will be described in more detail below in combination with examples. Those familiar with the present technology can easily understand other advantages and effects of the present invention from the contents disclosed in this specification.

[0028] The inventors of the present invention unexpectedly discovered a new fluorine-substituted pyrimidinedione compound by modifying the structure of Elagolix through extensive practical research. The compound has better biological activity and more ideal pharmacokinetic properties than Elagolix and other compounds commonly used in the prior art. Therefore, the inventors aimed to provide a new compound that can be a gonadotropin-releasing hormone (GnRH) receptor antagonist, and completed the present invention based on this.

[0029] In a first aspect, the present invention provides a compound or a pharma- ceutically acceptable salt, isomer, prodrug, polymorph or solvate thereof, the chemical structure of which is shown in Formula I:

[0030] [ka]

[0031] In the present invention, the term "salt" should be understood as any form of active compound used in the present invention. The compound may be in ionic form, may carry a charge, may be bound to a counterion (cation or anion), or may be in solution. The definition of "salt" may also include quaternary ammonium salts and complex compounds of active molecules with other molecules and ions, particularly complex compounds through ionic interactions. The definition of "salt" particularly includes physiologically acceptable salts (this term can be interpreted as equivalent to "pharmaceutical acceptable salts").

[0032] In the present invention, the term "pharmaceutical acceptable salt" generally means any salt that is physiologically tolerable (usually meaning harmless, especially when used as a counter ion) when used in a suitable manner for treatment (especially when applied or used in humans and / or mammals). These physiologically acceptable salts can be formed with cations or bases. And, in the context of the present invention, particularly when administered to humans and / or mammals, these physiologically acceptable salts should be interpreted as salts formed with at least one compound provided according to the present invention (usually an acid (deprotonated), e.g. an anion) and at least one physiologically tolerable cation (preferably an inorganic cation). In the context of the present invention, specifically, salts formed with alkali metals and alkaline earth metals, and salts formed with ammonium cation (NH4 + ) may include salts formed with sodium (mono) or (di) salts, potassium (mono) or (di) salts, magnesium or calcium. These physiologically acceptable salts may also be formed with anions or acids. In the context of the present invention, particularly when administered to humans and / or mammals, these physiologically acceptable salts should be understood as salts formed with at least one compound provided according to the present invention (usually protonated (e.g., on nitrogen), e.g., cations) and at least one physiologically tolerable anion. In the context of the present invention, specifically, salts formed with physiologically tolerable acids, i.e., salts formed with a particular active compound and a physiologically tolerable organic or inorganic acid, may include salts formed with hydrochloric acid, hydrobromic acid, sulfuric acid, methanesulfonic acid, formic acid, acetic acid, oxalic acid, succinic acid, malic acid, tartaric acid, mandelic acid, fumaric acid, lactic acid or citric acid, but are not limited thereto. Exemplary pharma- ceutically acceptable salts of compounds of formula I have the following chemical structures:

[0033] [ka]

[0034] The compounds of the present invention represented by formula I above may include enantiomers determined by the chiral centers present or isomers determined by the double bonds present (e.g., Z, E). All single isomers, enantiomers, diastereomers or cis-trans isomers and mixtures thereof are within the scope of the present invention.

[0035] The term "prodrug" in the present invention is used in the broadest sense and includes derivatives that can be converted to the compounds of the present invention in the body. Methods for preparing prodrugs of a given active compound should be known to those skilled in the art. See, for example, Krogsgaard-Larsen et al., Textbook of Drug Design and Discovery, 1999, pp. 111-115, 2001. See the relevant disclosures in "The Art of Drug design and Discovery" (Taylor & Francis, April 2002).

[0036] In the present invention, the term "solvate" generally means that any form of active compound according to the present invention is combined with another molecule (usually a polar solvent) by non-covalent bond, and the substances obtained thereby specifically include, but are not limited to, hydrates and alcoholates (e.g., methylates).

[0037] In a second aspect, the present invention provides a method for preparing a compound as provided in the first aspect of the present invention, the method comprising hydrolyzing a compound of formula 1-12 to prepare a compound of formula I. The reaction scheme is as follows:

[0038] [ka]

[0039] In the preparation method provided by the present invention, the hydrolysis reaction can usually be carried out under conditions in which a base is present. Those skilled in the art can select an appropriate type and amount of base to be used in the hydrolysis reaction. For example, the base can be an alkali metal hydroxide or the like, more specifically, lithium hydroxide or the like. In addition, for example, the amount of the base used is usually approximately equal to or in excess of the compound of formula 1-12. Specifically, the molar ratio of the compound of formula 1-12 to the base may be 1:1.4-1.6.

[0040] In the preparation method provided by the present invention, the reaction may be generally carried out under conditions ranging from room temperature to the boiling point of the reaction solvent, preferably at room temperature. Those skilled in the art may appropriately adjust the reaction time of the hydrolysis reaction according to the progress of the reaction. Methods for monitoring the progress of the reaction should be known to those skilled in the art. For example, analytical methods such as chromatography and nuclear magnetic resonance spectroscopy may be used. In addition, the specific reaction time may be, for example, 0.5 to 1 hour, 1 to 1.5 hours, 1.5 to 2 hours, 2 to 3 hours, 3 to 4 hours, or a longer reaction time.

[0041] In the preparation method provided by the present invention, the reaction is usually carried out in the presence of a solvent. In addition, the solvent should be a good solvent for the reaction raw materials and contain water so that the reaction raw materials can be sufficiently dispersed to ensure smooth progress of the reaction. The type and amount of a suitable reaction solvent should be known to those skilled in the art. For example, the reaction solvent may contain water or an ether solvent, etc., specifically, tetrahydrofuran, etc.

[0042] In the preparation methods provided by the present invention, those skilled in the art can select a suitable method for post-treatment of the reaction product, which may include, for example, high performance chromatography preparation, freeze-drying, etc.

[0043] In a third aspect, the present invention provides the use of the compound provided in the first aspect of the present invention or a pharma- ceutically acceptable salt, isomer, prodrug, polymorph or solvate thereof in the manufacture of a drug. The compound of the present invention can effectively inhibit the GnRH receptor, and therefore can be a gonadotropin releasing hormone receptor antagonist, and can be used to treat sex hormone-related diseases. The gonadotropin releasing hormone receptor antagonist provided by the present invention can be applied to a wide range of therapeutic uses, and can be used to treat disease conditions related to various sex hormones in men, women, and general mammals (also referred to as "individuals" in the present invention). Specific examples of the sex hormone-related disease include endometriosis, menopause, menstrual irregularities, uterine myoma, uterine fibroids, polycystic ovary syndrome, endometriosis, uterine leiomyoma, lupus erythematosus, hirsutism, precocious puberty, dwarfism, acne, alopecia, gonadal hormone-dependent tumors (e.g., prostate cancer, uterine cancer, breast cancer, ovarian cancer, pituitary gonadotropic cell adenoma, etc.), pituitary adenoma producing gonadotropic hormones, sleep apnea syndrome, irritable bowel syndrome, premenstrual syndrome, prostatic hyperplasia, contraception and infertility, Alzheimer's disease, and the like.

[0044] In a fourth aspect, the present invention provides a pharmaceutical composition comprising a compound as provided in the first aspect of the present invention or a pharma- ceutically acceptable salt, isomer, prodrug, polymorph or solvate thereof, which may further comprise at least one pharma- ceutically acceptable vector.

[0045] In the present invention, the composition may include one or more pharma- ceutically acceptable vectors. The vector generally refers to a vector used for administering a therapeutic agent, which does not induce the production of antibodies harmful to an individual receiving the composition, and does not have excessive toxicity after administration. These vectors are well known to those skilled in the art. For example, Remington's Pharmaceutical Sciences (Mack Pub. Co., NJ, 1991) discloses information on pharma- ceutically acceptable vectors. Specifically, the vector may include one or more combinations of saline, buffer, glucose, water, glycerin, ethanol, adjuvants, etc., but is not limited thereto.

[0046] In the pharmaceutical composition provided by the present invention, the compound may be a single active ingredient, or may be combined with other active ingredients to form a combination drug. The other active ingredients may be various other drugs that can be used to treat diseases related to gonadal hormones. Usually, the content of the active ingredient in the composition is a safe and effective amount. The safe and effective amount should be adjustable by those skilled in the art. For example, the dosage of the compound and the active ingredient of the pharmaceutical composition usually depends on the patient's weight, the type of application, and the condition and severity of the disease. For example, the dosage of the compound as an active ingredient may usually be 0.1-10 mg / kg / day, 0.1-0.5 mg / kg / day, 0.5-1 mg / kg / day, 1-2 mg / kg / day, 2-3 mg / kg / day, 3-4 mg / kg / day, 4-5 mg / kg / day, 5-6 mg / kg / day, 6-8 mg / kg / day, or 8-10 mg / kg / day, more preferably 0.5-5 mg / kg / day.

[0047] The compounds provided by the present invention can be adapted to any type of administration, and may be administered orally or non-digestively. For example, they may be administered via pulmonary, nasal, rectal and / or intravenous injection, and more specifically, may be administered intradermally, subcutaneously, intramuscularly, intraarticularly, intraperitoneally, pulmonary, oral, sublingually, nasally, transdermally, vaginally, or orally or non-digestively. Those skilled in the art may select an appropriate formulation form depending on the administration method. For example, formulation forms suitable for oral administration may include, but are not limited to, pills, tablets, chewable agents, capsules, granules, drops or syrups. For example, formulation forms suitable for non-digestive administration may include, but are not limited to, solutions, suspensions, rehydratable dry formulations or sprays. For example, those suitable for rectal administration may usually be suppositories.

[0048] In a fifth aspect, the present invention provides a method of treatment comprising administering to an individual a therapeutically effective amount of a compound as provided in the first aspect of the invention, or a pharmaceutical composition as provided in the fourth aspect of the invention.

[0049] In the present invention, an "individual" generally includes humans, non-human primates, and mammals (e.g., dogs, cats, horses, sheep, pigs, cows, etc.) that can benefit from treatment with the formulation, reagent kit, or combination.

[0050] In the present invention, the term "therapeutically effective amount" generally means a dose capable of achieving the effect of treating the above-listed diseases after a suitable administration period.

[0051] As a GnRHR antagonist, the compound provided by the present invention has activity equivalent to or superior to elagolix in calcium flow detection experiments. In addition, in pharmacokinetic studies, the absolute bioavailability of the compound provided by the present invention is obviously higher than that of elagolix. In plasma protein binding studies, the protein binding rate of the compound provided by the present invention in SD rat plasma and healthy human plasma was slightly higher than that of elagolix. Furthermore, the protein binding was not dependent on the drug concentration. As is clear from the above, the compound provided by the present application or its pharmacologic acceptable salt, isomer, prodrug, crystalline polymorph, or solvate has better biological activity and more ideal pharmacokinetic properties than other drugs of the same kind in the prior art, and has good industrialization potential.

[0052] The present invention will be further described below with reference to examples, but the scope of the present invention is not limited thereto.

[0053] Example 1 The specific preparation route of compound G201 in the examples was as follows:

[0054] [ka]

[0055] Compound 1-1 (10 g, 72.4 mmol), pyridine (200 mL), and selenium dioxide (16 g, 144.8 mmol) were added to a three-neck flask. This was reacted at 100°C for 2 hours while protecting with nitrogen gas. After the reaction was completed, the mixture was filtered with suction, and the filtrate was centrifuged. 1N hydrochloric acid was added, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated saline, dried over anhydrous sodium sulfate, filtered with suction, and centrifuged to obtain off-white solid 1-2 (12 g).

[0056] Compound 1-2 (12 g, 71.4 mmol) and methanol (200 mL) were added to a three-neck flask. Then, thionyl chloride (17 g, 142.8 mmol) was added dropwise at 0°C, and the mixture was stirred at room temperature overnight. After the reaction was completed, the methanol was centrifuged and a saturated aqueous solution of sodium bicarbonate was added, followed by extraction with ethyl acetate. The organic phase was washed with saturated saline, dried over anhydrous sodium sulfate, filtered under suction, and centrifuged to obtain a pale yellow oily substance 1-3 (9 g, 49.4 mmol).

[0057] Compound 1-3 (4 g, 20.4 mmol), anhydrous tetrahydrofuran (60 mL) and (R)-tert-butylsulfinamide (2.47 g, 20.4 mmol) were added to a three-neck flask. Next, while protecting with nitrogen gas, tetraisopropyl orthotitanate (11.6 g, 40.8 mmol) was added and refluxed and stirred for 6 hours. After the reaction was completed, water was added to perform a quench reaction, and ethyl acetate was added to dilute the mixture and then filtered by suction. The filtrate was separated and the aqueous phase was extracted with ethyl acetate. The organic phases were then combined, washed with saturated saline, dried over anhydrous sodium sulfate, filtered by suction and centrifuged to obtain a yellow oily substance 1-4 (6 g, 19.2 mmol).

[0058] Compound 1-4 (6 g, 19.2 mmol) and methanol (60 mL) were added to a three-neck flask. Then, at 0°C, sodium borohydride (1.1 g, 28.7 mmol) was added in several portions and reacted at room temperature for 3 hours. After the reaction was completed, water was added to quench the reaction, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated saline, dried over anhydrous sodium sulfate, suction filtered, centrifuged, and purified by column chromatography (petroleum ether / ethyl acetate = 10:1 to 5:1) to obtain a yellow oily substance 1-5 (3 g, 9.5 mmol).

[0059] Compound 1-5 (3 g, 9.5 mmol) and anhydrous tetrahydrofuran (40 mL) were added to a three-neck flask. Then, lithium aluminum hydride (0.43 g, 11.4 mmol) was added in several portions at 0°C, and the mixture was allowed to react at room temperature for 2 hours. After the reaction was completed, water was added to quench the reaction, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated saline, dried over anhydrous sodium sulfate, and filtered by suction. It was then centrifuged and purified by column chromatography (petroleum ether / ethyl acetate = 10:1 to 3:1) to obtain a yellow oily substance 1-6 (1.5 g, 5.8 mmol).

[0060] Compound 1-6 (1.5 g, 5.8 mmol), ethyl acetate (10 mL) and hydrochloric acid gas / ethyl acetate (10 mL) were added to a flask. This was reacted at room temperature for 1 hour. After the reaction was completed, the mixture was centrifuged to obtain a yellow solid 1-7 (1.5 g), which was then directly added to the next step.

[0061] Compound 1-7 (1.5 g), dichloromethane (20 mL), triethylamine (2.9 g, 28.9 mmol) and di-tert-butyl dicarbonate (1.5 g, 6.9 mmol) were added to a flask. This was reacted at room temperature for 16 hours. After the reaction was completed, the mixture was centrifuged and purified by column chromatography (petroleum ether / ethyl acetate = 15:1 to 10:1) to obtain a yellow solid 1-8 (1.3 g, 5.1 mmol).

[0062] Compound 1-8 (1.3 g, 5.1 mmol), compound 1-A (1.9 g, 5.1 mmol), triphenylphosphine (2.0 g, 7.6 mmol), anhydrous tetrahydrofuran (30 mL) and diethyl azodicarboxylate (1.5 g, 7.6 mmol) were added to a three-neck flask. This was refluxed and reacted for 16 hours. After the reaction was completed, the mixture was centrifuged and purified by column chromatography (petroleum ether / ethyl acetate = 10:1 to 5:1) to obtain a white solid 1-9 (2.1 g, 3.4 mmol).

[0063] Compound 1-9 (2.1 g, 3.4 mmol), compound 1-B (0.7 g, 4.1 mmol), sodium carbonate (1.4 g, 13.6 mmol), dioxane (27 mL), water (9 mL) and tetrakis(triphenylphosphine)palladium (0.4 g, 0.34 mmol) were added to a three-neck flask. This was reacted at 90 ° C for 16 hours while protecting with nitrogen gas. After the reaction was completed, it was filtered by suction, and the filtrate was centrifuged and purified by column chromatography (petroleum ether / ethyl acetate = 15: 1 to 5: 1) to obtain a light yellow solid 1-10 (1.35 g, 2.0 mmol).

[0064] Compound 1-10 (250 mg, 0.38 mmol), dichloromethane (2 mL) and trifluoroacetic acid (2 mL) were added to a flask and reacted at room temperature for 1 hour. After completion of the reaction, the mixture was centrifuged to obtain a yellow solid 1-11 (210 mg).

[0065] The crude product, compound 1-11 (210 mg), acetonitrile (3 mL), compound 1-C (220 mg, 1.13 mmol), and potassium carbonate (156 mg, 1.13 mmol) were added to a flask and reacted for 16 hours at 80° C. After completion of the reaction, the mixture was centrifuged and subjected to preparative thin layer chromatography (petroleum ether / ethyl acetate=1.5 / 1) to obtain a colorless oily substance, 1-12 (210 mg, 0.31 mmol).

[0066] Compound 1-12 (210 mg, 0.31 mmol), tetrahydrofuran (3 mL), water (2 mL) and lithium hydroxide (20 mg, 0.47 mmol) were added to a flask and reacted at room temperature for 2 hours. After the reaction was completed, the mixture was centrifuged, high-pressure adjusted (acetonitrile 10-55 / 7 min), and freeze-dried to obtain a white solid G201 (20 mg).

[0067] 1HNMR CD3ODδ:7.62~7.64(m,1H), 7.39~7.57(m,4H), 7.11~7.23(m,4H), 6.65~6.80(m,1H), 5.38~5.49(m,2H), 4.81( q,1H), 4.46~4.53(m,2H), 3.90(s,3H), 2.81~2.84(m,2H), 2.37~2.40(m,2H), 2.12(d,3H), 1.79~1.83(m,2H).

[0068] LC-MS: m / z=649.8(M+1), LCMS purity >98%.

[0069] Example 2 Calcium flow detection: Using the FLIPR calcium flow detection reagent kit (Calcium 4 assay kit), changes in calcium were measured in the cells of recombinant human gonadotropin releasing hormone receptor (GnRHR) stable cell line CHO-K1 / GNRHR / Gα15. GnRHR is a type of G protein-coupled receptor (GPCRs), and GPCRs signals release calcium in the cells through the Gq pathway. Therefore, by detecting calcium release in the cells using a calcium ion-sensitive fluorescent probe, it is possible to detect functional changes in GnRHR that have transmitted signals through the Gq pathway. The experimental procedure was as follows.

[0070] 1. The cells were seeded into a 384-well microplate (Corning, Cat#: 3764) at 10,000 cells / well and 20 μl / well using F12+10% FBS medium, and cultured overnight (18 hours).

[0071] 2. Probenecid stock solution (500mM) and Calcium 4 stock solution were mixed at 1:100 to prepare the dye working solution. In addition, the test samples G201 and Elagolix (2mM in DMSO) were diluted with assay buffer. That is, 5μl of the 2mM stock solution was diluted to 200μl (50μM) to prepare the 5X sample working solution of the first concentration. Next, 20μl of the first sample working solution was diluted to 200μl and mixed uniformly to obtain the sample working solution of the second concentration. In this way, the sample working solutions diluted in 8 steps were prepared. Then, 20μl / well of the dye working solution and 10μl / well of the sample working solution were added in sequence to the cells cultured overnight, and the cells were then cultured in the cell culture device for 45min. After that, the cells were kept dark at room temperature and equilibrated for 15min.

[0072] A 3.5X EC80 agonist working solution was made: 5 μl of 1 mM Buserelin stock solution was diluted to 200 μl (25 μM). 71 μl of the 25 μM Buserelin working solution was then diluted to 10 μl (0.179 μM) with assay buffer.

[0073] 4. According to the position of the cells in the 384-well microplate, agonist working solution was added to the sample plate at 40 μl / well for FLIPR detection.

[0074] Test results: G201 was able to inhibit intracellular calcium release in a dose-dependent manner in CHO-K1 / GNRHR / Gα15 cells. 50 The GnRHR antagonist activity of G201 was 37.59 nM compared with 45.73 nM for elagolix. These results indicate that G201 is a GnRHR antagonist with activity equivalent to or superior to that of elagolix.

[0075] Example 3 Pharmacokinetic study in rats: Twelve female SD rats were divided into four groups (three rats per group) and administered G201 and Elagolix Sodium intravenously and intragastrically (G201 was dissolved in saline containing 5% N,N-dimethylacetamide and 5% polyoxyethylene castor oil EL, and Elagolix Sodium was dissolved in saline as is). The doses were 25 mg / kg and 50 mg / kg, respectively. The animals were fasted overnight before administration and fed again 4 h after administration. Water was allowed ad libitum throughout the entire test period. Blood samples were collected from the retroorbital venous plexus before administration and 5, 15, and 30 min, 1, 2, 4, 6, 8, and 24 h after administration. The volume of blood collected was approximately 0.2 mL. All blood samples were centrifuged at 4000 rpm for 10 min within 1 h to separate the upper layer of plasma. The samples were then placed in a refrigerator within 1 h and stored at -20°C until analysis. Plasma samples were subjected to 1:8 methanol protein precipitation, and the blood drug concentrations of G201 and elagolix were detected by LC-MS / MS, and the main pharmacokinetic parameters were calculated using DAS3.2.7 software.

[0076] The results showed that the mean CL of G201 administered intravenously was 1.62 L / h / kg, and the mean t 1 / 2 The mean CL and Vz of elagolix sodium were 0.79h and 1.82L / kg, respectively. The mean CL and Vz of elagolix sodium were 2.48L / h / kg and 8.46L / kg, respectively. The apparent volume of distribution of G201 was also significantly smaller than that of elagolix, indicating reduced tissue distribution.

[0077] When G201 was orally administered to rats, it was rapidly absorbed and max is 0.42h, mean AUC 0-∞ is 16351.7h *ng / mL, which was three times that of elagolix sodium. The absolute bioavailability of G201 when administered orally was 51.92%, which was clearly higher than that of elagolix sodium (23.65%). See Figure 1 for the drug concentration-time curves when G201 / elagolix was administered intravenously, see Figure 2 for the drug concentration-time curves when administered orally, and see Table 1 for pharmacokinetic parameters.

[0078] [Table 1]

[0079] Example 4 Pharmacokinetic studies in mice: Ten female ICR mice were divided into two groups, five mice per group, and G201 was administered intravenously and orally by gavage. The dose was 10 mg / kg. Blood samples were collected from the retroorbital venous plexus 0.083 h, 0.25 h, 0.5 h, 1 h, 2 h, 4 h, 6 h, and 8 h after administration. The blood volume was approximately 0.1 mL. Within 1 h, the blood samples were centrifuged at 4000 rpm for 10 min to separate the upper layer of plasma. Within 1 h, the samples were stored in a refrigerator at -80°C and awaited measurement. Before blood collection, the test animals were fasted for at least 12 h, but were not deprived of water. In addition, the animals were fasted and deprived of water during the blood collection process, and allowed to eat and drink freely 2 h after administration. The blood drug concentration of G201 in the ICR mice was detected by LC-MS / MS, and the AUC 0-t , AUC 0-∞ , C max , t 1 / 2 The main pharmacokinetic parameters, including those listed above, were calculated.

[0080] The results showed that when G201 was administered intravenously to mice, the average CL was 3.18 L / h / kg and the average t 1 / 2 The mean time to absorption was 0.26 h and the mean Vz was 1.17 L / kg. G201 was rapidly absorbed when orally administered to mice, and the T max is 0.27h, mean AUC0-∞ is 1266.3h * ng / mL. The absolute bioavailability was 38.9%, which was obviously higher than that of Elagolix (the absolute bioavailability of Elagolix was 10% when administered at 10 mg / kg to mice, according to literature reports). See Figure 3 for the blood concentration-time curves of G201 when administered intravenously and orally to ICR mice, and see Table 2 for the pharmacokinetic parameters.

[0081] [Table 2]

[0082] Example 5 Plasma protein binding assay: Plasma from previously cultured SD rats (five males and five females) or healthy humans was collected and thoroughly mixed with working solutions containing different concentrations of the test substances (G201 (1 μM, 10 μM, 100 μM) and Elagolix (10 μM)) and a control (warfarin sodium (10 μM)). 50 μL of the drug-containing plasma sample was then added to an equal volume of blank buffer to obtain an unfiltered sample. Meanwhile, 350 μL was transferred to the ultrafiltration device (the inner tube of the ultrafiltration tube) and centrifuged at 37 °C (10,000 g x 3 minutes). After the centrifugation was completed, 50 μL of blank plasma was added to the sample (filtrate sample) taken out of the outer tube. Similarly, an equal volume of blank buffer was added to the sample (filtered excess sample) taken out of the inner tube. Then, 2 minutes after mixing, the internal standard solution was added. All samples were rotated for 10 minutes to centrifuge precipitated proteins, and the supernatant was obtained. The concentrations of the test substance and the control were then measured by LC-MS / MS, and the free percentage (free percentage = (drug concentration in filtrate sample / drug concentration in unfiltered plasma) × 100%) and bound percentage (bound percentage = 100% - free percentage) were calculated. In addition, the recovery rate (%) = (drug concentration in filtrate sample × volume + drug concentration in filtrate sample × volume) / drug concentration in unfiltered plasma × total volume × 100.

[0083] The results showed that the protein binding rates of elagolix in SD rat plasma and healthy human plasma at a drug concentration of 10 μM were 87.71% and 87.25%, respectively, which were consistent with literature reports. On the other hand, the protein binding rates of G201 in SD rat plasma and healthy human plasma were 89.79% and 89.94%, respectively, which were slightly higher than that of elagolix. In addition, there was no dependency of protein binding on drug concentration in the range of 1 to 100 μM.

[0084] [Table 3]

[0085] Example 6 Efficacy study in castrated male cynomolgus monkeys: Three male cynomolgus monkeys aged 3-5 years (body weight 3.8-4.0 kg) (purchased from Guangxi Guidong Primate Development Experimental Co., Ltd.) were obtained, fasted overnight, and anesthetized to undergo bilateral orchiectomy. Serum luteinizing hormone (LH) was then measured in the cynomolgus monkeys 3-7 weeks after castration by radioimmunoassay. As a result, serum LH in the male cynomolgus monkeys was obviously elevated (about 10-fold) 3 weeks after castration, and LH levels were almost stable 7 weeks after castration.

[0086] After the castration surgery model was successfully created, it was divided into two administration cycles. In the two administration cycles, G201 and elagolix sodium (solvent: 0.5% CMC-Na) were administered orally once by gavage. The doses were 10 mg / kg and 30 mg / kg, respectively, and the administration volume was 10 mL / kg for both. Each administration cycle was administered after a 2-week elution period. During each administration cycle, blood was collected before administration and 0.25 h, 0.5 h, 1 h, 2 h, 3 h, 6 h, 8 h, and 24 h after administration, and the serum was separated to detect the sex hormone LH.

[0087] The results showed that after oral administration of G201 and elagolix sodium to castrated cynomolgus monkeys, serum LH levels were significantly decreased. In both cases, the levels were lowest 3 h after administration and then increased again. Animals in the 30 mg / kg elagolix sodium group returned to pre-administration levels 24 h after administration. Animals in the 10 mg / kg G201 group showed a slightly higher post-administration change than those in the 30 mg / kg elagolix sodium group. During the study, all animals were in good general condition, had stable body weights, and no abnormalities related to the test substances were observed.

[0088] In conclusion, a single oral administration of G201 significantly decreased serum LH levels in castrated male cynomolgus monkeys, and the inhibitory effect at a dose of 10 mg / kg was comparable to or superior to that of elagolix sodium at 30 mg / kg.

[0089] As described above, the present invention overcomes various drawbacks in the prior art and has a high degree of industrial applicability.

[0090] The above embodiments are merely illustrative of the principles and effects of the present invention, and are not intended to limit the present invention. Those skilled in the art can supplement or modify the above embodiments without departing from the spirit and scope of the present invention. Therefore, any equivalent supplement or modification that a person skilled in the art can complete without departing from the spirit and technical ideas disclosed in the present invention is also included in the scope of the claims of the present invention.

Claims

1. A compound having a chemical structural formula represented by Formula I, or a pharma- ceutically acceptable salt, enantiomer, diastereomer, cis-trans isomer or stereoisomer, crystal or solvate thereof. 【Chemistry 1】

2. A process for preparing the compound of claim 1, comprising the steps of: The preparation method includes hydrolyzing a compound of formula 1-12 to prepare a compound of formula I, the reaction scheme being as follows: 【Chemistry 2】

3. The preparation method according to claim 2, characterized in that the hydrolysis reaction can be carried out in the presence of a base.

4. 13. Use of the compound according to claim 1 or a pharma- ceutically acceptable salt, enantiomer, diastereomer, cis-trans isomer or stereoisomer, crystal or solvate thereof in the manufacture of a medicament.

5. The use according to claim 4, characterized in that the drug is selected from gonadotropin releasing hormone receptor antagonists.

6. The use according to claim 4, characterized in that the drug is selected from drugs for treating diseases related to gonadal hormones.

7. The use according to claim 6, characterized in that the disease associated with gonadal hormones is selected from endometriosis, menopause, menstrual irregularities, uterine myoma, uterine fibroids, polycystic ovary syndrome, uterine leiomyoma, lupus erythematosus, hirsutism, precocious puberty, dwarfism, acne, alopecia, gonadal hormone-dependent tumors, pituitary adenomas producing gonadotropic hormones, sleep apnea syndrome, irritable bowel syndrome, premenstrual syndrome, prostatic hyperplasia, contraception and infertility, and Alzheimer's disease.

8. The use according to claim 7, characterized in that the gonadal hormone-dependent tumor is selected from prostate cancer, uterine cancer, breast cancer, ovarian cancer and pituitary gonadotroph adenoma.

9. A pharmaceutical composition comprising the compound of claim 1 or a pharma- ceutically acceptable salt, enantiomer, diastereomer, cis-trans isomer or stereoisomer, crystal or solvate thereof.

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