The thienopyrimidine dione carboxylic acid compound is substituted at position 5 by a condensed polycyclic structure.

Abstract

The invention relates to a series of thienopyrimidine dione carboxylic acid compounds substituted at position 5 by a condensed polycyclic structure. Specifically, the invention relates to a compound of formula (II) and its pharmaceutical salt.

Description

Polycyclic substituted 5-carboxylic acid thienopyrimidine dione compounds and their applications

[0001] The present invention claims the following priority:

[0002] CN202110204744.5, application date February 23, 2021;

[0003] CN202110523923.5, application date May 13, 2021;

[0004] CN202210125821.2, application date February 10, 2022. Technical Field

[0005] The present invention relates to a series of polycyclic substituted 5-carboxylic acid thienopyrimidinedione compounds and applications thereof, and particularly to compounds represented by formula (II) and pharmaceutically acceptable salts thereof. Background Art

[0006] Endometriosis is the presence of endometrial-like tissue outside the uterus. While not fatal, its symptoms, including chronic pelvic pain and dysmenorrhea, can cause significant distress in patients, including pain and infertility. It is extremely difficult to cure and prone to recurrence. Furthermore, clinical medications for this condition have drawbacks such as long duration of use, numerous side effects, and inconvenient dosing. According to the World Bank's 2017 population estimates, approximately 190 million women worldwide suffer from endometriosis.

[0007] The pathogenesis of endometriosis is still unclear, and current clinical treatment options focus on either controlling estrogen levels, controlling inflammation, or both. For example, first-line treatments include nonsteroidal anti-inflammatory drugs (NSAIDs) or oral contraceptives, while second-line treatments include oral aromatase inhibitors, danazol, or injectable gonadotropin-releasing hormone (GnRH) agonists. However, oral contraceptives have a non-response rate of approximately one-third to one-quarter of patients, progestins have the side effect of increasing obesity, and are particularly contraindicated in patients seeking pregnancy. Aromatase inhibitors have side effects such as cardiac toxicity and abnormal lipid metabolism, and gonadotropin-releasing hormones have perimenopausal side effects. Polypeptide GnRH receptor agonist or antagonist compounds present numerous challenges, such as oral absorption, dosage form, dosage volume, drug stability, sustained action, and metabolic stability.

[0008] Small molecule compounds can be administered orally, which is convenient and fast, with obvious advantages. GnRH receptor antagonists competitively bind to GnRH receptors, block the binding of GnRH to the receptors, directly inhibit the hypothalamic-pituitary-ovarian axis, and then inhibit the secretion of follicle-stimulating hormone and luteinizing hormone, reduce estrogen levels, have a rapid onset of action, and have few side effects. At present, in addition to the first to be listed Elagolix, the small molecule GnRH receptor antagonist is at the forefront of research and development. In December 2020, the FDA approved the second small molecule oral antagonist Relugolix for the first indication of treating advanced prostate cancer. The indication for the treatment of uterine fibroids has been approved in Japan, and endometriosis has entered Phase III clinical trials. The third antagonist Linzagolix is ​​in Phase III clinical trials for the treatment of endometriosis and uterine fibroids.

[0009]

[0010] Although a large number of meaningful clinical trials have been conducted in these areas, there is still a need to continue researching and developing more effective small molecule GnRH receptor antagonists.

[0011] Summary of the Invention

[0012] The present invention provides a novel polycyclic GnRH receptor antagonist, which has significant inhibitory activity on GnRH receptors.

[0013] The present invention provides a compound represented by formula (II) or a pharmaceutically acceptable salt thereof,

[0014]

[0015] in,

[0016] L1 and L2 are each independently selected from -(CH2) n -;

[0017] L3 and L4 are independently selected from -CH2-, -CH=CH-, -O- and -S-;

[0018] R1 and R2 are independently selected from H, OH, F, Cl, C 1-3 Alkyl and C 1-3 Alkoxy, the C 1-3 Alkyl and C 1-3 The alkoxy groups are each independently optionally substituted with 1, 2 or 3 R a replace;

[0019] Alternatively, R1 and R2 together with the carbon atom to which they are attached form C 3-6 Cycloalkyl or 4-6 membered heterocycloalkyl, the C 3-6 Cycloalkyl or 4-6 membered heterocycloalkyl is optionally substituted by 1, 2 or 3 Rb replace;

[0020] R3, R4, R5, R6, R7, R8 and R9 are independently selected from H, F, Cl, Br, I, OH, NH2, CN, C 1-3 Alkyl, C 1-3 Alkoxy and C 3-6 Cycloalkyl, the C 1-3 Alkyl, C 1-3 Alkoxy and C 3-6 The cycloalkyl group is optionally substituted by 1, 2 or 3 R c replace;

[0021] n is selected from 0, 1 and 2;

[0022] Each R a 、R b and R c are independently selected from H, F, Cl, Br, I, NH2 and OH.

[0023] In some embodiments of the present invention, R1 and R2 are independently selected from H, OH, F and CH3, and CH3 is optionally substituted by 1, 2 or 3 Fs. Other variables are as defined in the present invention.

[0024] In some embodiments of the present invention, R1 and R2 are independently selected from H, and other variables are as defined in the present invention.

[0025] In some embodiments of the present invention, the above R1 and R2 together with the atoms to which they are commonly attached form a cyclopropyl, cyclobutyl, cyclopentyl, oxetanyl or azetidinyl group, wherein the cyclopropyl, cyclobutyl, cyclopentyl, oxetanyl or azetidinyl group is optionally replaced by 1, 2 or 3 R b Substitution, other variables are as defined in the present invention.

[0026] In some embodiments of the present invention, the above-mentioned R1 and R2 together with the atoms to which they are commonly attached form cyclopropyl, cyclobutyl, cyclopentyl, oxetanyl and azetidinyl, and the cyclopropyl, cyclobutyl, cyclopentyl, oxetanyl and azetidinyl are optionally substituted with 1, 2 or 3 F, and other variables are as defined in the present invention.

[0027] In some embodiments of the present invention, the above R1 and R2 together with the atoms to which they are attached form Other variables are as defined in the present invention.

[0028] In some embodiments of the present invention, R3, R4, R5 and R6 are independently selected from H, F, Cl, OH, NH2, CN, CH3, CF3, OCH3 and OCF3, and other variables are as defined in the present invention.

[0029] In some embodiments of the present invention, R3, R4, R5 and R6 are independently selected from H and F, and other variables are as defined in the present invention.

[0030] In some embodiments of the present invention, the above structural unit Selected from Other variables are as defined in the present invention.

[0031] In some embodiments of the present invention, R7, R8 and R9 are independently selected from H, F, Cl, OH, NH2, CN, CH3, CF3, OCH3 and OCF3, and other variables are as defined in the present invention.

[0032] In some embodiments of the present invention, R7, R8 and R9 are independently selected from H and F, and other variables are as defined in the present invention.

[0033] In some embodiments of the present invention, the above structural unit Selected from Other variables are as defined in the present invention.

[0034] In some embodiments of the present invention, the above structural unit Selected from Other variables are as defined in the present invention.

[0035] In some embodiments of the present invention, L3 and L4 are independently selected from O, and other variables are as defined in the present invention.

[0036] In some embodiments of the present invention, the above n is selected from 1 and 2, and other variables are as defined in the present invention.

[0037] In some embodiments of the present invention, the above structural unit Selected from Other variables are as defined in the present invention.

[0038] In some embodiments of the present invention, the above structural unit Selected from Other variables are as defined in the present invention.

[0039] In some embodiments of the present invention, the above structural unit Selected from Other variables are as defined in the present invention.

[0040] In some embodiments of the present invention, the above structural unit Selected from Other variables are as defined in the present invention.

[0041] In some embodiments of the present invention, the above-mentioned compound or a pharmaceutically acceptable salt thereof is selected from the group consisting of:

[0042]

[0043] wherein L1, L2, R1, R2, R3, R4, R5 and R6 are as defined in the present invention.

[0044] The present invention also provides a compound represented by formula (I) or a pharmaceutically acceptable salt thereof,

[0045]

[0046] in,

[0047] L1 and L2 are each independently selected from -(CH2) n -;

[0048] L3 and L4 are independently selected from -CH2-, -CH=CH-, -O- and -S-;

[0049] R1 and R2 are independently selected from H, OH, F, Cl, C 1-3 Alkyl and C 1-3 Alkoxy, the C 1-3 Alkyl and C 1-3 The alkoxy group is optionally substituted with 1, 2 or 3 R a replace;

[0050] Alternatively, R1 and R2 together with the atoms to which they are attached form a C 3-6 Cycloalkyl or 4-6 membered heterocycloalkyl, the C 3-6 Cycloalkyl or 4-6 membered heterocycloalkyl is optionally substituted by 1, 2 or 3 R b replace;

[0051] R3, R4, R5 and R6 are independently selected from H, F, Cl, Br and I;

[0052] n is selected from 0, 1 and 2;

[0053] Each R a and R b Each independently selected from H, F, Cl, Br, I;

[0054] The "4-6 membered heterocycloalkyl" contains 1, 2 or 3 heteroatoms selected from N, NH, O and S.

[0055] Some other solutions of the present invention are obtained by any combination of the above variables.

[0056] The present invention also provides the following compounds or pharmaceutically acceptable salts thereof:

[0057]

[0058] The present invention also provides the use of the above compound or a pharmaceutically acceptable salt thereof in the preparation of drugs related to GnRH receptor antagonists.

[0059] In some embodiments of the present invention, the above-mentioned GnRH receptor antagonist-related drug is a drug used to prevent and / or treat endometriosis and / or uterine fibroid-related diseases.

[0060] Technical Effects

[0061] The compound of the present invention has a significant inhibitory effect on the human gonadotropin-releasing hormone receptor, can significantly inhibit the growth of the volume of endometriosis lesions in mice, and has excellent efficacy; PK results show that the compound of the present invention has a high exposure in plasma, a low clearance rate, a long half-life, and a high oral bioavailability, exhibiting excellent pharmacokinetic properties, and is a good molecule for development of oral administration.

[0062] Definition and Description

[0063] Unless otherwise indicated, the following terms and phrases used herein are intended to have the following meanings. A particular term or phrase should not be construed as indefinite or unclear unless specifically defined, but rather should be understood in accordance with its ordinary meaning. When a trade name appears in this document, it is intended to refer to the corresponding commercial product or its active ingredient.

[0064] The term "pharmaceutically acceptable" as used herein refers to those compounds, materials, compositions and / or dosage forms which, within the scope of sound medical judgment, are suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response or other problems or complications, commensurate with a reasonable benefit / risk ratio.

[0065] The term "pharmaceutically acceptable salt" refers to salts of the compounds of the present invention, prepared by reacting the compounds of the present invention with relatively nontoxic acids or bases. When the compounds of the present invention contain relatively acidic functional groups, base addition salts can be obtained by contacting the compounds with a sufficient amount of base in neat solution or in a suitable inert solvent. Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amine, or magnesium salts, or similar salts. When the compounds of the present invention contain relatively basic functional groups, acid addition salts can be obtained by contacting the compounds with a sufficient amount of acid in neat solution or in a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include inorganic acid salts such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, bicarbonate, phosphoric acid, monohydrogen phosphate, dihydrogen phosphate, sulfuric acid, bisulfate, hydroiodic acid, phosphorous acid, and the like; and organic acid salts such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, and methanesulfonic acid; and salts of amino acids (such as arginine) and organic acids such as glucuronic acid. Certain specific compounds of the present invention contain both basic and acidic functional groups and can be converted into either base or acid addition salts.

[0066] Pharmaceutically acceptable salts of the present invention can be synthesized by conventional chemical methods from parent compounds containing acid radicals or bases. Generally, such salts are prepared by reacting these compounds in free acid or base form with a stoichiometric amount of a suitable base or acid in water or an organic solvent or a mixture of the two.

[0067] The compounds of the present invention may exist in specific geometric or stereoisomeric forms. The present invention contemplates all such compounds, including cis- and trans-isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)-isomers, (L)-isomers, and racemic mixtures and other mixtures thereof, such as enantiomerically or diastereomerically enriched mixtures, all of which are within the scope of the present invention. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers and mixtures thereof are encompassed within the scope of the present invention.

[0068] Unless otherwise indicated, the term "enantiomer" or "optical isomer" refers to stereoisomers that are mirror images of one another.

[0069] Unless otherwise indicated, the term "cis-trans isomers" or "geometric isomers" arises from the inability to rotate freely about double bonds or single bonds forming ring carbon atoms.

[0070] Unless otherwise indicated, the term "diastereomer" refers to stereoisomers that have two or more chiral centers and that are not mirror images of each other.

[0071] Unless otherwise indicated, "(+)" indicates dextrorotatory, "(-)" indicates levorotatory, and "(±)" indicates racemic.

[0072] Unless otherwise specified, use a solid wedge key. and dotted wedge key To indicate the absolute configuration of a stereocenter, use a straight solid bond and straight dashed bond Indicate the relative configuration of stereocenters with a wavy line Indicates a wedge-shaped solid key or dotted wedge key Or use a wavy line Indicates a straight solid bond and straight dashed bond

[0073] Unless otherwise specified, when a compound contains a double bond structure, such as a carbon-carbon double bond, a carbon-nitrogen double bond, or a nitrogen-nitrogen double bond, and each atom on the double bond is connected to two different substituents (in a double bond containing a nitrogen atom, a lone pair of electrons on the nitrogen atom is considered as a substituent to which it is connected), if a wavy line is used between the atom on the double bond and its substituent in the compound, When connected, it represents the (Z) isomer, (E) isomer or a mixture of the two isomers of the compound. For example, the following formula (A) represents that the compound exists in the form of a single isomer of formula (A-1) or formula (A-2) or in the form of a mixture of the two isomers of formula (A-1) and formula (A-2); the following formula (B) represents that the compound exists in the form of a single isomer of formula (B-1) or formula (B-2) or in the form of a mixture of the two isomers of formula (B-1) and formula (B-2). The following formula (C) represents that the compound exists in the form of a single isomer of formula (C-1) or formula (C-2) or in the form of a mixture of the two isomers of formula (C-1) and formula (C-2).

[0074]

[0075] Unless otherwise indicated, the term "tautomer" or "tautomeric form" refers to isomers of different functional groups that are in dynamic equilibrium at room temperature and readily interconvert into each other. If tautomerism is possible (e.g., in solution), chemical equilibrium of the tautomers can be achieved. For example, proton tautomers (also called prototropic tautomers) include interconversions via proton migration, such as keto-enol isomerization and imine-enamine isomerization. Valence tautomers include interconversions via reorganization of some bonding electrons. A specific example of keto-enol tautomerization is the interconversion between pentane-2,4-dione and 4-hydroxypent-3-en-2-one.

[0076] Unless otherwise indicated, the terms "enriched in one isomer", "isomerically enriched", "enriched in one enantiomer" or "enantiomerically enriched" mean that the content of one isomer or enantiomer is less than 100%, and the content of that isomer or enantiomer is greater than or equal to 60%, or greater than or equal to 70%, or greater than or equal to 80%, or greater than or equal to 90%, or greater than or equal to 95%, or greater than or equal to 96%, or greater than or equal to 97%, or greater than or equal to 98%, or greater than or equal to 99%, or greater than or equal to 99.5%, or greater than or equal to 99.6%, or greater than or equal to 99.7%, or greater than or equal to 99.8%, or greater than or equal to 99.9%.

[0077] Unless otherwise indicated, the term "isomer excess" or "enantiomeric excess" refers to the difference between the relative percentages of two isomers or two enantiomers. For example, if the content of one isomer or enantiomer is 90% and the content of the other isomer or enantiomer is 10%, the isomer or enantiomeric excess (ee value) is 80%.

[0078] The compounds of the present invention may contain unnatural proportions of atomic isotopes at one or more of the atoms that constitute the compound. For example, the compounds may be labeled with radioactive isotopes, such as tritium ( 3 H), iodine-125( 125 I) or C-14( 14 C). For example, deuterated drugs can be formed by replacing hydrogen with heavy hydrogen. The bond between deuterium and carbon is stronger than the bond between ordinary hydrogen and carbon. Compared with non-deuterated drugs, deuterated drugs have advantages such as reduced toxic side effects, increased drug stability, enhanced efficacy, and prolonged drug biological half-life. All isotopic variations of the compounds of the present invention, whether radioactive or not, are included within the scope of this invention.

[0079] The terms "optional" or "optionally" mean that the subsequently described event or circumstance may but need not occur, and that the description includes instances where said event or circumstance occurs and instances where it does not.

[0080] The term "substituted" means that any one or more hydrogen atoms on a particular atom are replaced by a substituent, which may include deuterium and hydrogen variants, as long as the valence state of the particular atom is normal and the substituted compound is stable. When the substituent is oxygen (i.e., =O), it means that two hydrogen atoms are replaced. Oxygen substitution does not occur on aromatic groups. The term "optionally substituted" means that it may be substituted or unsubstituted, and unless otherwise specified, the type and number of substituents can be any on the basis of chemical achievable.

[0081] When any variable (e.g., R) occurs more than once in a compound's composition or structure, its definition at each occurrence is independent. Thus, for example, if a group is substituted with 0-2 Rs, the group may be optionally substituted with up to two Rs, with each occurrence of R being an independent choice. Furthermore, combinations of substituents and / or their variants are permissible only if such combinations result in stable compounds.

[0082] When the number of a linking group is 0, such as -(CRR)0-, it means that the linking group is a single bond.

[0083] When one of the variables is selected from a single bond, it means that the two groups it connects are directly connected. For example, when L in ALZ represents a single bond, it means that the structure is actually AZ.

[0084] Unless otherwise specified, when a group has one or more connectable sites, any one or more sites of the group can be connected to other groups through chemical bonds. When the chemical bond connection mode is non-positional and there are H atoms at the connectable sites, when the chemical bond is connected, the number of H atoms at the site will decrease accordingly with the number of connected chemical bonds, and become a group with a corresponding valence. The chemical bond connecting the site to other groups can be a straight solid bond. Straight dotted key or wavy lines For example, the straight solid bond in -OCH3 indicates that it is connected to other groups through the oxygen atom in the group; The straight dashed bond in the group indicates that the two ends of the nitrogen atom in the group are connected to other groups; The wavy line in the phenyl group indicates that it is connected to other groups through the carbon atoms at positions 1 and 2 in the phenyl group; Indicates that any linkable site on the piperidinyl group can be connected to other groups through a chemical bond, including at least In these four connection methods, even if an H atom is drawn on -N-, Still includes For groups connected in this way, when one chemical bond is connected, the H at that site will be reduced by one and become a corresponding monovalent piperidine group. It means that the 3-carbon atom in the cyclohexyl group is connected to other groups through a double bond.

[0085] Unless otherwise specified, the term “C 1-3 "Alkyl" itself or in combination with other terms refers to a straight or branched chain saturated hydrocarbon group consisting of 1 to 3 carbon atoms. 1-3 Alkyl groups include C 1-2 and C 2-3 Alkyl, etc.; it can be monovalent (such as methyl), divalent (such as methylene) or polyvalent (such as methine). 1-3 Examples of alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), and the like.

[0086] Unless otherwise specified, the term “C 1-3 "Alkoxy" by itself or in combination with other terms refers to an alkyl group containing 1 to 3 carbon atoms, which is attached to the rest of the molecule through an oxygen atom. 1-3 Alkoxy groups include C 1-2 、C 2-3 , C3 and C2 alkoxy, etc. 1-3 Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy (including n-propoxy and isopropoxy), and the like.

[0087] Unless otherwise specified, “C 3-6 "Cycloalkyl" means a saturated monocyclic hydrocarbon group consisting of 3 to 6 carbon atoms, wherein the C 3-6 Cycloalkyl groups include C 3-5 、C 4-5 and C 5-6 Cycloalkyl, etc.; it may be monovalent, divalent or polyvalent. 3-6 Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like.

[0088] Unless otherwise specified, the term "4-6 membered heterocycloalkyl" by itself or in combination with other terms means a saturated monocyclic radical consisting of 4 to 6 ring atoms, 1, 2, 3 or 4 of which are heteroatoms independently selected from O, S and N, and the remainder being carbon atoms, wherein the nitrogen atom is optionally quaternized and the nitrogen and sulfur heteroatoms are optionally oxidized (i.e., NO and S(O) p, p is 1 or 2). In addition, with respect to the "4-6 membered heterocycloalkyl", a heteroatom may occupy the position at which the heterocycloalkyl is attached to the rest of the molecule. The 4-6 membered heterocycloalkyl includes 5-6 membered, 4 membered, 5 membered and 6 membered heterocycloalkyls, etc. Examples of 4-6 membered heterocycloalkyl groups include, but are not limited to, azetidinyl, oxetanyl, thietanyl, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, tetrahydrothiophenyl (including tetrahydrothiophen-2-yl and tetrahydrothiophen-3-yl, etc.), tetrahydrofuranyl (including tetrahydrofuran-2-yl, etc.), tetrahydropyranyl, piperidinyl (including 1-piperidinyl, 2-piperidinyl and 3-piperidinyl, etc.), piperazinyl (including 1-piperazinyl and 2-piperazinyl, etc.), morpholinyl (including 3-morpholinyl and 4-morpholinyl, etc.), dioxanyl, dithianyl, isoxazolidinyl, isothiazolidinyl, 1,2-oxazinyl, 1,2-thiazinyl and hexahydropyridazinyl, etc.

[0089] Unless otherwise specified, C n-n+m or C n -C n+m Any specific case including n to n+m carbons, such as C 1-12 Including C1, C2, C3, C4, C5, C6, C7, C8, C9, C 10 、C 11 , and C 12 , also includes any range from n to n+m, such as C 1-12 Including C 1- 3. C 1-6 、C 1-9 、C 3-6 、C 3-9 、C 3-12 、C 6-9 、C 6-12 , and C 9-12 Similarly, n-membered to n+m-membered means that the number of atoms in the ring is n to n+m, for example, a 3-12-membered ring includes a 3-membered ring, a 4-membered ring, a 5-membered ring, a 6-membered ring, a 7-membered ring, an 8-membered ring, a 9-membered ring, a 10-membered ring, an 11-membered ring, and a 12-membered ring, and also includes any range from n to n+m, for example, a 3-12-membered ring includes a 3-6-membered ring, a 3-9-membered ring, a 5-6-membered ring, a 5-7-membered ring, a 6-7-membered ring, a 6-8-membered ring, and a 6-10-membered ring, etc.

[0090] The compounds of the present invention can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining them with other chemical synthesis methods, and equivalent substitutions well known to those skilled in the art. Preferred embodiments include but are not limited to the examples of the present invention.

[0091] The structures of the compounds of the present invention can be confirmed by conventional methods well known to those skilled in the art. If the present invention relates to the absolute configuration of the compounds, the absolute configuration can be confirmed by conventional techniques in the art. For example, single crystal X-ray diffraction (SXRD) is used to collect diffraction intensity data on the cultured single crystal using a Bruker D8 venture diffractometer, using CuKα radiation as the light source and scanning mode: After scanning and collecting relevant data, the crystal structure is further analyzed using the direct method (Shelxs97) to confirm the absolute configuration.

[0092] The present invention uses the following abbreviations: DMSO stands for dimethyl sulfoxide; MeOH stands for methanol; ACN stands for acetonitrile; DEA stands for diethylamine; CO2 stands for carbon dioxide; psi stands for pounds-force per square inch; Ac stands for acetyl, Ph stands for phenyl, DMAC stands for N,N-dimethylacetamide; Solutol stands for polyethylene glycol-15 hydroxystearate; and PEG stands for polyethylene glycol.

[0093] The solvents used in the present invention can be obtained commercially. Software naming, commercially available compounds use supplier catalog names. DETAILED DESCRIPTION

[0094] The present invention is described in detail below by way of examples, but is not intended to limit the present invention in any way. While the present invention has been described in detail herein, and specific embodiments thereof have been disclosed, it will be apparent to those skilled in the art that various changes and modifications can be made to the specific embodiments of the present invention without departing from the spirit and scope of the present invention.

[0095] Reference Example 1: Intermediate BB-1

[0096]

[0097] first step

[0098] To a solution of compound B-1 (dimethyl maleate, 50 g, 346.92 mmol) and methyl thioglycolate (37.01 g, 348.68 mmol) in tetrahydrofuran (300 ml) was added piperidine (886.18 mg, 10.41 mmol), and the mixture was stirred at 25°C for 2 hours. After completion of the reaction, as monitored by thin-layer chromatography (petroleum ether:ethyl acetate = 5:1), 300 ml of water was added, and the mixture was extracted twice with 200 ml of ethyl acetate each time. The combined organic phases were washed with 200 ml of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to dryness to provide compound B-2.

[0099] 1 H NMR (400MHz, CDCl3) δ=3.89-3.81(m,1H),3.76(s,3H),3.75(d,3H),3.69(s,3H),3.55-3.32(m,2H),3.00(m,1H),2.73(m,1H).

[0100] Step 2

[0101] Sodium (2.78 g, 120.92 mmol) was added to methanol (10 ml) at a temperature below 20°C, and the mixture was stirred at a temperature below 64°C until the solid dissolved. This was then prepared into a sodium methoxide methanol solution, cooled to 20°C, and added to a solution of B-2 (10 g, 39.96 mmol) in tetrahydrofuran (20 ml). The reaction mixture was stirred at 66°C under nitrogen for 3 hours. Isopropyl ether (100 ml) and acetic acid (1 ml) were added, the mixture was cooled to 20°C, filtered, and the filter cake was added to a mixture of phosphoric acid (10 ml) and water (20 ml). The mixture was extracted twice with ethyl acetate (20 ml each). The combined organic phases were washed with 30 ml of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain compound B-3.

[0102] Step 3

[0103] A mixture of compound B-3 (7.4 g, 33.91 mmol), pyridine (4.03 g, 50.89 mmol), and hydroxylamine hydrochloride (2.47 g, 35.57 mmol) was stirred at 50°C for 2 hours. 2 ml of phosphoric acid and 20 ml of water were added to the reaction solution, and the mixture was extracted twice with 20 ml of ethyl acetate each time. The organic layers were combined, washed with 20 ml of saturated sodium bicarbonate and once with 20 ml of saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain compound B-4.

[0104] Step 4

[0105] To a solution of compound B-4 (5.2 g, 22.29 mmol) in acetic acid (5 ml), 4 mol ethyl acetate (52.00 ml) was added, and the reaction mixture was stirred at 25°C for 12 hours. The reaction mixture was filtered, and the filter cake was concentrated under reduced pressure to dryness to obtain the hydrochloride salt of compound B-5.

[0106] MS-ESI calculated value [M+H-MeOH] + 184.0, measured value 184.1.

[0107] Step 5

[0108] To a solution of compound B-5 (2 g, 7.95 mmol, hydrochloride) in tetrahydrofuran (20 ml) and water (10 ml) was added potassium carbonate (1.65 g, 11.92 mmol), and phenyl chloroformate (2.49 g, 15.89 mmol) was added dropwise at 5-10°C. The reaction solution was stirred at 5-10°C for 1 hour. 50 ml of water was added to the reaction solution, and the mixture was extracted twice with ethyl acetate, 50 ml each time. The organic phases were combined, washed with 50 ml of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to dryness. Ethyl acetate (5 ml) and petroleum ether (50 ml) were added, and the mixture was beaten at 30°C for 30 minutes. The mixture was filtered and the filter cake was dried under reduced pressure to obtain compound BB-1.

[0109] MS-ESI calculated value [M+H] + 336.1, measured value 336.1.

[0110] Reference Example 2: Intermediate BB-2

[0111]

[0112] first step

[0113] To a solution of compound B-6 (3-bromopropanol, 5 g, 35.97 mmol) and 4-dimethylaminopyridine (439.49 mg, 3.6 mmol) in dichloromethane (25 ml) was added dropwise a dichloromethane solution (5 ml) of acetic anhydride (4.04 g, 39.57 mmol). The mixture was warmed to 25°C and stirred at 25°C for 4 hours. The reaction solution was washed with 1 mol / L hydrochloric acid (10 ml x 2), the aqueous phase was collected and extracted with dichloromethane (30 ml x 3), the combined organic phases were washed with saturated sodium bicarbonate aqueous solution (10 ml x 2), washed with saturated brine (10 ml x 2), the collected organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain intermediate compound BB-2.

[0114] 1 H NMR (400MHz, CDCl3) δ = 4.18-4.25 (m, 2H), 3.44-3.51 (m, 2H), 2.15-2.23 (m, 2H), 2.07 (s, 3H).

[0115] Example 1

[0116]

[0117]

[0118] first step

[0119] To a solution of compound 1a (3,4-difluoroanisole, 40 g, 277.5 mmol) in tetrahydrofuran (400 mL) was added lithium diisopropylamide (166.53 mL, 2 mol / L) dropwise at -70°C. The reaction system was stirred at -70°C for 0.5 hour. A solution of N,N-dimethylformamide (25.62 mL, 333.06 mmol) in tetrahydrofuran (24 mL) was added dropwise at -70°C to -60°C. The reaction system was stirred at -70°C for 1 hour. Acetic acid (25 mL) and water (100 mL) were added to the reaction system at -65°C. The mixture was extracted with ethyl acetate (200 mL x 3). The combined organic phases were washed sequentially with water (100 mL x 3) and saturated brine (100 mL x 3). The organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to yield compound 1b.

[0120] Step 2

[0121] To a solution of compound 1b (10 g, 58.10 mmol) in dichloromethane (100 ml) at -20°C, boron tribromide (29.11 g, 116.19 mmol) was added dropwise. The mixture was slowly warmed to 25°C and stirred at 25°C for 12 hours. Methanol (200 ml) and water (100 ml) were added dropwise to the reaction system, and the mixture was warmed to 40°C and stirred at 40°C for 2 hours. The layers were separated, the aqueous phase was extracted with dichloromethane (300 ml x 2), and the combined organic phases were extracted with aqueous sodium hydroxide solution (1 mol / L, 400 ml x 3). The extract was acidified to pH 2-3 with concentrated hydrochloric acid and extracted with ethyl acetate (300 ml x 3). The organic phases were collected, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain compound 1c.

[0122] 1 H NMR (400MHz, CDCl3) δ = 11.13 (s, 1H), 10.29 (s, 1H), 7.37 (q, J = 9.3Hz, 1H), 6.77-6.68 (m, 1H).

[0123] Step 3

[0124] To a solution of compound 1c (1.5 g, 9.49 mmol) in N,N-dimethylformamide (20 ml) were added sodium iodide (284.42 mg, 1.90 mmol) and potassium carbonate (1.97 g, 14.23 mmol). The mixture was stirred at 25°C for 0.5 hours. Compound BB-2 (2.06 g, 11.39 mmol) was then added, and the mixture was warmed to 60°C and stirred at 60°C for 12 hours. The reaction system was poured into 30 ml of water and extracted with ethyl acetate (50 ml x 5). The combined organic phases were washed with water (20 ml x 5) and then washed once with saturated brine. The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to provide compound 1d.

[0125] 1 H NMR(400MHz, CDCl3)δ=10.45-10.38(m,1H),7.37-7.28(m,1H),6.74-6.65(m, 1H), 4.28 (t, J = 6.2Hz, 2H), 4.16-4.13 (m, 2H), 2.22-2.16 (m, 2H), 2.06 (s, 3H).

[0126] Step 4

[0127] To a solution of compound 1d (3.25 g, 12.59 mmol) in tetrahydrofuran (30 mL) was added an aqueous solution (3 mL) of sodium borohydride (490 mg, 12.95 mmol) at 0°C, and the reaction system was stirred at 0°C for 0.5 hours. Water (10 mL) was added to the reaction system at 0°C, and the mixture was extracted with ethyl acetate (30 mL x 3). The combined organic phases were washed with water (10 mL x 2) and saturated brine (10 mL x 3). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to provide compound 1e.

[0128] Step 5

[0129] To a solution of compound 1e (2.65 g, 10.18 mmol) and 5-fluoro-2-hydroxybenzaldehyde (1.57 g, 11.2 mmol) in tetrahydrofuran (20 ml) was added tri-n-butylphosphine (3.71 g, 18.33 mmol). The mixture was stirred for 0.1 hour, and then a solution of azodicarbonyldipiperidine (4.62 g, 18.33 mmol) in tetrahydrofuran (5 ml) was added dropwise at 0°C. The mixture was warmed to 25°C and stirred at 25°C for 12 hours. The reaction system was poured into 10 ml of water and extracted with ethyl acetate (30 ml x 3). The combined organic phases were washed with water (10 ml x 3) and brine (10 ml x 3). The organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by column chromatography (silica gel, petroleum ether:ethyl acetate = 1 / 0 to 20 / 1) to provide compound 1f.

[0130] 1 H NMR (400MHz, CDCl3) δ = 10.31 (d, J = 3.2Hz, 1H), 7.49 (dd, J = 3.2, 8.4Hz, 1H), 7.33-7.27 (m, 1H), 7.23-7.18 (m, 1H), 7.18-7.11 ( m,1H),6.72-6.57(m,1H),5.25(d,J=2.8Hz,2H),4.19(t,J=6.2Hz,2H),4.06(t,J=6.2Hz,2H),2.09-2.06(m,2H),2.04(s,3H).

[0131] Step 6

[0132] To a solution of compound 1f (1.04 g, 2.72 mmol) in dichloromethane (10 ml) at 0°C was added m-chloroperbenzoic acid (1.66 g, 85% purity, 8.16 mmol). The mixture was warmed to 25°C and stirred at 25°C for 12 hours. 2 ml of saturated aqueous sodium sulfite solution was added to the reaction solution, followed by 10 ml of water, and the mixture was extracted with ethyl acetate (30 ml x 3). The combined organic phases were washed with water (10 ml x 2) and saturated brine (10 ml x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The crude product was purified by preparative chromatography (petroleum ether:ethyl acetate = 3:1) to afford compound 1g.

[0133] 1 H NMR (400MHz, CDCl3) δ = 8.19 (s, 1H), 7.18-7.08 (m, 2H), 7.00-6.93 (m, 1H), 6.89 (dd, J = 3.2, 8.4Hz, 1H), 6.63- 6.57(m,1H),5.14-5.09(m,2H),4.23(t,J=6.2Hz,2H),4.04(t,J=6.2Hz,2H),2.14-2.09(m,2H),2.06(s,3H).

[0134] Step 7

[0135] To a 10 mL solution of compound 1g (1 g, 2.51 mmol) in methanol was added an aqueous solution of potassium hydroxide (1 mL, 20% purity, 489.03 μmol), and the mixture was stirred at 25°C for 6 hours. The reaction solution was poured into 10 mL of water and then extracted with ethyl acetate (30 mL x 3). The combined organic phases were washed with water (10 mL x 2) and saturated brine (10 mL x 2). The organic phases were collected, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The crude product was purified by column chromatography (silica gel, petroleum ether:ethyl acetate = 5:1 to 3:1) to provide compound 1h.

[0136] 1 H NMR (400MHz, CDCl3) δ = 7.15 (q, J = 9.2Hz, 1H), 7.00 (dd, J = 5.2, 8.8Hz, 1H), 6.91 (s, 1H), 6.69-6.62 (m, 2H),6.53(dt,J=3.0,8.6Hz,1H),5.17(d,J=2.0Hz,2H),4.22(t,J=5.8Hz,2H),3.88(q,J=5.0Hz,2H), 2.49(br s,1H),2.15-2.05(m,2H).

[0137] Step 8

[0138] To a solution of compound 1h (421 mg, 1.28 mmol) in tetrahydrofuran (400 mL) at 0°C was added sodium hydroxide (135.04 mg, 60% purity, 3.38 mmol), and the mixture was stirred at 0°C for 0.5 hours. A solution of p-toluenesulfonyl chloride (244.50 mg, 1.28 mmol) in tetrahydrofuran (5 mL) was added dropwise to the reaction system at 0°C, and the mixture was stirred at 25°C for 12 hours. 10 mL of water was added to the reaction solution, and the mixture was extracted with ethyl acetate (30 mL x 3). The combined organic phases were washed with water (10 mL x 2), saturated brine (10 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The crude product was purified by preparative chromatography (petroleum ether:ethyl acetate = 3:1) to provide compound 1i.

[0139] Step 9

[0140] To a solution of compound 1i (53 mg, 170.82 μmol) in acetic acid (1 mL) was added nitric acid (1.46 mL, 60% purity, 19.51 mmol) dropwise at 80°C, and the mixture was stirred at 80°C for 2 hours. The reaction solution was poured into 40 mL of ice water, and the pH was adjusted to 7 with saturated aqueous sodium bicarbonate. The aqueous phase was extracted with ethyl acetate (30 mL x 5), and the combined organic phases were washed with water (30 mL x 3) and saturated brine (20 mL x 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to provide compound 1j.

[0141] 1 H NMR (400MHz, CDCl3) δ = 7.92 (d, J = 7.6Hz, 1H), 7.14-7.10 (m, 1H), 6.76-6.73 (d, J = 12.0Hz, 1H), 6. 57-6.12(m,1H),5.20(d,J=1.2Hz,2H),4.48-4.42(m,2H),4.37-4.31(m,2H),2.17-2.14(m,2H).

[0142] Step 10

[0143] To a solution of compound 1j (46 mg, 129.48 μmol) in ethyl acetate (10 mL) was added wet palladium on carbon (10 mg, 10% purity), and the atmosphere was replaced with hydrogen three times. The mixture was stirred at 24°C under a hydrogen atmosphere (15 psi) for 12 hours. The reaction mixture was filtered through celite, and the filtrate was concentrated to provide compound 1k.

[0144] 1 H NMR (400MHz, CDCl3) δ = 7.17-7.07 (m, 2H), 6.82-6.78 (m, 1H), 6.76 (d, J = 11.6Hz, 1H), 6.68 (d, J = 9.0Hz, 1H), 6.63-6 .56(m,1H),5.07(d,J=2.0Hz,2H),4.35-4.31(m,2H),4.16-4.11(m,2H),2.13-2.09(m,2H); LC-MS: m / z=326.1[M+H] + .

[0145] Step 11

[0146] To a solution of compound 1k (41 mg, 78.41 μmol) in tetrahydrofuran (3 mL) were added compound BB-1 (26.29 mg, 78.41 μmol) and triethylamine (7.93 mg, 78.41 μmol). The mixture was stirred at 70°C for 10 hours. The reaction solution was poured into 10 mL of water and extracted with ethyl acetate (30 mL x 5). The combined organic phases were washed with water (10 mL x 3) and saturated brine (10 mL x 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and purified by preparative chromatography (petroleum ether:ethyl acetate = 2 / 1) to provide compound 1l.

[0147] 1 H NMR (400MHz, CDCl3) δ = 8.86 (s, 1H), 7.97 (s, 1H), 7.81 (d, J = 8.0Hz, 1H), 7.16-7.06 (m, 1H), 6.84 (d, J = 11.6Hz, 1H), 6.78 (m, 1H), 6.57 (br s,1H),5.16(d,J=1.6Hz,2H),4.36(t,J=5.2Hz,2H),4.22(t,J=5.2Hz,2H),3.91(s,3H),3.90(s,3H),2.11(m,2H); LC-MS: m / z=567.1[M+H] + .

[0148] Step 12

[0149] To a solution of compound 11 (22 mg, 28.57 μmol, 73.57% purity) in tetrahydrofuran (2 ml) and methanol (1 ml) was added an aqueous solution of lithium hydroxide monohydrate (5.99 mg, 142.85 μmol) (1 ml), and the mixture was stirred at 26°C for 2 hours. 1 mol / L dilute hydrochloric acid was added to the reaction solution to adjust the pH to approximately 6, followed by extraction with ethyl acetate (5 ml x 5). The combined organic phases were washed with water (5 ml x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated. The crude product was purified by preparative chromatography (dichloromethane:methanol = 10 / 1) to provide compound 1.

[0150] 1H NMR(400MHz,DMSO-d6)δ=12.05-11.76(br s,1H),7.44-7.32(m,1H),7.29(s,1H),7.25(d,J=7.8Hz,1H),7.16(d,J=12.0Hz,1 H),7.08-7.00(m,1H),5.75(s,1H),5.14-5.09(m,1H),5.09-5.04(m,1H),4.48(br t, J=4.8Hz, 2H), 4.30 (br t, J=4.6Hz, 2H), 2.01-1.91 (m, 2H); LC-MS: m / z=521.1[M+H] + .

[0151] Example 2

[0152]

[0153] first step

[0154] To a solution of compound 2a (1,1-cyclopropane dimethanol, 10 g, 97.91 mmol) in dichloromethane (20 ml) was added dropwise a hydrobromic acid acetic acid solution (72.00 g, 293.65 mmol, 33% purity) at 0-5°C, and the mixture was stirred at 10-20°C for 2 hours. 100 ml of water was added, and the mixture was extracted twice with 20 ml of dichloromethane each time. The organic phases were combined and washed twice with saturated sodium bicarbonate each time, 50 ml each time. The organic phase was washed with 50 ml of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to dryness to obtain compound 2b.

[0155] Step 2

[0156] To a solution of 2,3-difluoro-6-hydroxybenzaldehyde (5 g, 31.63 mmol) in N,N-dimethylformamide (50 mL) were added potassium carbonate (6.57 g, 47.55 mmol), sodium iodide (948.08 mg, 6.33 mmol), and compound 2b (9.82 g, 47.44 mmol). The mixture was stirred at 60°C for 12 hours. 100 mL of water was added to the reaction solution, and the mixture was extracted twice with 50 mL of ethyl acetate each time. The combined organic phases were washed twice with 50 mL of water each time, then with 50 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to dryness under reduced pressure. The mixture was purified by column chromatography (silica gel, 100-200 mesh, petroleum ether:ethyl acetate = 1:0 to 10:1) to obtain compound 2c.

[0157] Step 3

[0158] To a solution of compound 2c (4.5 g, 15.83 mmol) in tetrahydrofuran (40 mL) was added a solution of sodium borohydride (0.83 g, 21.94 mmol) in water (5 mL) at 0-5°C. The reaction mixture was stirred at 25°C for 1 hour. 50 mL of water was added to the reaction mixture, and the mixture was extracted twice with 20 mL of ethyl acetate each time. The combined organic phases were washed with 10 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to dryness to provide compound 2d.

[0159] Step 4

[0160] To a solution of compound 2d (4.4 g, 15.37 mmol) in dichloromethane (40 mL) at 0-5°C were added pyridine (3.65 g, 46.11 mmol) and thionyl chloride (3.66 g, 30.74 mmol). The reaction was stirred at 25°C under nitrogen for 12 hours. 50 mL of water was added to the reaction solution, and the mixture was extracted twice with 50 mL of ethyl acetate each time. The organic layers were combined, washed with 20 mL of saturated sodium bicarbonate and once with 30 mL of saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give compound 2e.

[0161] Step 5

[0162] To a solution of 5-fluoro-2-hydroxybenzaldehyde (1.84 g, 13.13 mmol) and compound 2e (4.0 g, 13.13 mmol) in acetonitrile (40 mL) were added potassium carbonate (2.72 g, 19.69 mmol) and sodium iodide (196.77 mg, 1.31 mmol). The reaction mixture was stirred at 60°C under nitrogen for 12 hours. The reaction mixture was filtered, and the filtrate was added with 50 mL of water and extracted twice with 50 mL of ethyl acetate each time. The organic phases were combined, washed with 50 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to dryness. Compound 2f was purified by silica gel column chromatography (100-200 mesh, petroleum ether:ethyl acetate = 1:0 to 20:1) to obtain compound 2f.

[0163] Step 6

[0164] To a solution of compound 2f (2.2 g, 5.39 mmol) in dichloromethane (25 mL) was added m-chloroperbenzoic acid (1.20 g, 5.93 mmol, 85% purity) at 0-5°C, and the reaction mixture was stirred at 30°C for 36 hours. The reaction mixture was filtered, and the filtrate was added with 20 mL of saturated aqueous sodium bicarbonate solution. The mixture was extracted twice with 20 mL of dichloromethane each time. The organic phases were combined, washed twice with 20 mL of saturated sodium bicarbonate each time, and then washed with 20 mL of saturated brine. The mixture was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to dryness to provide compound 2g.

[0165] Step 7

[0166] Potassium carbonate (2.15 g, 15.55 mmol) was added to a solution of compound 2g (2.2 g, 5.18 mmol) in methanol (10 ml) and water (2 ml). The reaction mixture was stirred at 40°C for 2 hours. 30 ml of water was added to the reaction mixture, and the mixture was extracted twice with 20 ml of ethyl acetate each time. The organic layers were combined, washed with 30 ml of saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to dryness under reduced pressure. Compound 2h was then purified by silica gel column chromatography (100-200 mesh, petroleum ether:ethyl acetate = 20:1 to 5:1) to obtain compound 2h.

[0167] Step 8

[0168] To a solution of compound 2h (1.63 g, 4.60 mmol) in tetrahydrofuran (30 mL) at 0°C were added sodium hydride (552.04 mg, 13.80 mmol, 60% purity) and p-toluenesulfonyl chloride (877.06 mg, 4.60 mmol). The reaction was stirred at 35°C under nitrogen for 60 hours. 2 mL of saturated aqueous ammonium chloride and 10 mL of water were added to the reaction solution, which was then extracted twice with 10 mL of ethyl acetate each time. The combined organic phases were washed with 10 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The mixture was purified by silica gel column chromatography (100-200 mesh, petroleum ether:ethyl acetate = 1:0 to 10:1) to provide compound 2i.

[0169] 1 H NMR (400MHz, CDCl3) δ = 7.19-7.06 (m, 2H), 6.75-6.67 (m, 3H), 5.17 (d, J = 1.6Hz, 2H), 4.13 (s, 2H), 3.98 (s, 2H), 0.72 (s, 4H).

[0170] Step 9

[0171] To a solution of compound 2i (360 mg, 1.07 mmol) in acetic acid (3 mL) was added dropwise nitric acid (518.84 mg, 5.35 mmol, 65% purity) at 60°C, and the reaction was stirred at 60°C for 2 hours. 10 mL of water was added to the reaction solution to precipitate a solid, which was filtered and dried under reduced pressure. The filter cake was purified by silica gel column chromatography (100-200 mesh, petroleum ether:ethyl acetate = 10:1 to 5:1) to provide compound 2j.

[0172] Step 10

[0173] To a solution of compound 2j (200 mg, 524.52 μmol) in ethanol (4 mL) and water (0.8 mL) at 79°C were added ammonium chloride (140.29 mg, 2.62 mmol) and reduced iron powder (146.46 mg, 2.62 mmol). The reaction was stirred at 79°C for 3 hours. Water (10 mL) was added to the reaction solution, and the mixture was extracted twice with 10 mL of ethyl acetate each time. The combined organic phases were washed with 10 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified by silica gel column chromatography (100-200 mesh, petroleum ether:ethyl acetate = 1:0 to 5:1) to provide compound 2k. LC-MS: m / z = 352.1 [M+H] + .

[0174] Step 11

[0175] To a solution of compound BB-1 (143.17 mg, 426.96 μmol) and compound 2k (150 mg, 426.96 μmol) in tetrahydrofuran (5 mL) was added triethylamine (43.2 mg, 426.96 μmol), and the reaction was stirred at 35°C for 60 hours. The reaction solution was concentrated to dryness under reduced pressure and purified by silica gel column chromatography (100-200 mesh, petroleum ether:ethyl acetate = 20:1 to 5:1) to provide compound 2l.

[0176] LC-MS: m / z = 593.2 [M+H] + .

[0177] Step 12

[0178] To a solution of compound 21 (100 mg, 168.77 μmol) in tetrahydrofuran (2 mL) and methanol (2 mL) was added a solution of lithium hydroxide monohydrate (35.41 mg, 843.83 μmol) in water (2 mL), and the reaction was stirred at 20°C for 2 hours. The reaction solution was adjusted to pH 4-5 with 1 M hydrochloric acid to precipitate a solid, which was filtered and the filter cake was concentrated under reduced pressure to dryness. Ethyl acetate (5 mL) and petroleum ether (25 mL) were added, and the mixture was stirred at 20°C for 30 minutes. The mixture was filtered and the filter cake was dried under reduced pressure to provide compound 2.

[0179] 1 H NMR (400MHz, DMSO-d6) δ=11.72(br s,1H),7.38(q,J=9.6Hz,1H),7.25(br d,J=8.0Hz,1H),7.16-7.03(m,2H),6.99(br s,1H),5.10(br s,2H),4.28(s,2H),4.10-4.03(m,2H),0.68-0.60(m,4H); LC-MS: m / z=547.1[M+H] + .

[0180] Example 3

[0181]

[0182] first step

[0183] To a solution of compound 3a (4-bromo-1-butanol, 10 g, 65.35 mmol) in dichloromethane (100 mL) were added triethylamine (9.92 g, 98.03 mmol) and acetic anhydride (7.34 g, 71.89 mmol) at 0-5°C. The mixture was stirred at 25°C under nitrogen for 12 hours. 100 mL of water was added to the reaction solution, and the mixture was extracted twice with 100 mL of dichloromethane each time. The combined organic phases were washed with 100 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to provide compound 3b.

[0184] Step 2

[0185] To a solution of 2,3-difluoro-6-hydroxymethylbenzaldehyde (4 g, 25.30 mmol) in N,N-dimethylformamide (40 ml) were added potassium carbonate (5.26 g, 38.04 mmol), sodium iodide (758.45 mg, 5.06 mmol), and compound 3b (6.42 g, 32.89 mmol). The mixture was stirred at 60°C for 12 hours. 100 ml of water was added to the reaction solution, and the mixture was extracted twice with 50 ml of ethyl acetate each time. The combined organic phases were washed twice with 50 ml of water each time, washed with 50 ml of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain compound 3c.

[0186] Step 3

[0187] To a solution of compound 3c (5 g, 18.37 mmol) in tetrahydrofuran (30 mL) was added a solution of sodium borohydride (800 mg, 21.15 mmol) in water (3 mL) at 0-5°C. The reaction mixture was stirred at 25°C for 1 hour. 50 mL of water was added to the reaction mixture, and the mixture was extracted twice with 20 mL of ethyl acetate each time. The combined organic phases were washed with 10 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to provide compound 3d.

[0188] Step 4

[0189] To a solution of 5-fluoro-2-hydroxybenzaldehyde (2.66 g, 18.96 mmol) and compound 3d (5.2 g, 18.96 mmol) in tetrahydrofuran (50 ml) at 0-5°C were added tri-n-butylphosphine (5.75 g, 28.44 mmol) and azodicarbonyldipiperidine (7.18 g, 28.44 mmol). The reaction was stirred at 25°C under nitrogen for 12 hours. The reaction solution was filtered, and the filtrate was added with 50 ml of water and extracted twice with 50 ml of ethyl acetate each time. The organic layers were combined, washed once with 50 ml of saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. Compound 3e was purified by silica gel column chromatography (100-200 mesh, petroleum ether:ethyl acetate = 1:0 to 20:1) to obtain compound 3e.

[0190] Step 5

[0191] To a solution of compound 3e (2.6 g, 6.56 mmol) in 30 mL of dichloromethane was added m-chloroperbenzoic acid (1.33 g, 6.56 mmol, 85% purity). The reaction was stirred at 25°C for 12 hours. The reaction mixture was filtered, and the filtrate was added with 30 mL of sodium bicarbonate. The mixture was extracted twice with 10 mL of dichloromethane. The combined organic phases were washed three times with 30 mL of saturated sodium bicarbonate, then washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to dryness. Compound 3f was then purified by silica gel column chromatography (100-200 mesh, petroleum ether:ethyl acetate = 1:0 to 10:1) to provide compound 3f.

[0192] Step 6

[0193] Potassium carbonate (1.08 g, 7.81 mmol) was added to a solution of compound 3f (1.5 g, 3.90 mmol) in methanol (2 ml) and water (2 ml). The reaction mixture was stirred at 25°C for 12 hours. 30 ml of water was added to the reaction mixture, and the mixture was extracted twice with 20 ml of ethyl acetate each time. The organic layers were combined, washed with 30 ml of saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to yield compound 3g.

[0194] 1 H NMR (400MHz, CDCl3) δ = 7.14 (q, J = 9.2Hz, 1H), 7.02-7.0 (m, 1H), 6.66-6.45 (m, 2H), 6.63-6.53 (m, 1H), 5. 11(d,J=1.8Hz,2H), 4.07(t,J=6.0Hz,2H), 3.70(t,J=6.0Hz,2H), 1.97-1.89(m,2H), 1.76-1.70(m,2H).

[0195] Step 7

[0196] To a solution of compound 3g (800 mg, 2.34 mmol) in tetrahydrofuran (20 mL) at 0°C were added sodium hydroxide (233.71 mg, 5.84 mmol, 60% purity) and p-toluenesulfonyl chloride (445.56 mg, 2.34 μmol). The reaction was stirred at 25°C under nitrogen for 60 hours. 2 mL of saturated aqueous ammonium chloride and 10 mL of water were added to the reaction solution, which was then extracted twice with 10 mL of ethyl acetate each time. The combined organic phases were washed with 10 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The mixture was purified by silica gel column chromatography (100-200 mesh, petroleum ether:ethyl acetate = 1:0 to 10:1) to provide compound 3h.

[0197] 1 H NMR (400MHz, CDCl3) δ = 7.17-7.10 (m, 2H), 6.70-6.62 (m, 3H), 5.04 (d, J = 2.0Hz, 2H), 4.16-4.13 (m, 4H), 2.10-2.04 (m, 4H).

[0198] Step 8

[0199] To a solution of compound 3h (100 mg, 308.36 μmol) in acetic acid (1 mL) was added nitric acid (149.46 mg, 1.54 mmol, 65% purity) at 0-5°C, and the reaction was stirred at 60°C for 1 hour. 10 mL of water was added to the reaction solution to precipitate a solid, which was filtered and the filter cake dried under reduced pressure. Ethyl acetate (1 mL) and petroleum ether (10 mL) were then added, and the mixture was stirred at 25°C for 30 minutes. The mixture was filtered and the filter cake dried under reduced pressure to provide compound 3i.

[0200] Step 9

[0201] To a solution of compound 3i (100 mg, 270.79 μmol) in ethyl acetate (10 mL) was added wet palladium carbon (10 mg, 10% purity, 50% water), and the reaction was stirred at 25° C. under 15 psi hydrogen for 7 hours. The reaction mixture was filtered and the filtrate was concentrated to give compound 3j.

[0202] LC-MS: m / z=340.1[M+H] + .

[0203] Step 10

[0204] To a solution of compound BB-1 (69.18 mg, 206.30 μmol) and compound 3j (70 mg, 206.30 μmol) in tetrahydrofuran (5 mL) was added triethylamine (41.75 mg, 412.60 μmol), and the reaction was stirred at 35°C for 36 hours. The reaction solution was concentrated to dryness under reduced pressure, and ethyl acetate (2 mL) and petroleum ether (20 mL) were added. The mixture was stirred at 25°C for 30 minutes, filtered, and the filter cake was dried under reduced pressure to provide compound 3k.

[0205] LC-MS: m / z = 581.3 [M+H] + .

[0206] Step 11

[0207] To a solution of compound 3k (150 mg, 258.39 μmol) in tetrahydrofuran (1 mL), methanol (1 mL), and water (1 mL) was added lithium hydroxide monohydrate (65.06 mg, 1.55 mmol), and the reaction was stirred at 25°C for 2 hours. The reaction mixture was adjusted to pH 4-5 with 1 M hydrochloric acid and stirred at 25°C for 10 minutes to precipitate a solid. The solid was filtered, and the filter cake was concentrated under reduced pressure to dryness. Ethyl acetate (5 mL) and petroleum ether (20 mL) were added, and the mixture was stirred at 25°C for 30 minutes. The mixture was filtered and dried under reduced pressure. The mixture was then purified by preparative HPLC (column: 3_Phenomenex Luna C18 75*30 mm*3 μm; mobile phase: [water (0.05% hydrochloric acid)-acetonitrile]; acetonitrile percentage: 47%-67%) to afford compound 3.

[0208] 1 H NMR(400MHz,DMSO-d6)δ=12.03(br s,1H),7.44(q,J=9.6Hz,1H),7.39-7.35(m,2H),7.17(d,J=11.2Hz,1H),6.95-6.88(m,1H),4.99(s,2H),4.17-4.14(m,4H),1.96(br s,4H); LC-MS: m / z=535.1[M+H] + .

[0209] Example 4

[0210]

[0211] first step

[0212] Compound 4a (3-bromopropanol, 12 g, 86.34 mmol) was dissolved in 60 ml of dichloromethane solution, and triethylamine (13.10 g, 129.50 μmol) was added and stirred at 25°C for 20 minutes. The reaction solution was cooled to 0°C, and then acetic anhydride (10.58 g, 103.60 mmol) was dissolved in 10 ml of dichloromethane and added dropwise to the reaction solution. The mixture was stirred at 25°C for 12 hours. The reaction solution was poured into 50 ml of water and the pH was adjusted to 3-5 with 1N dilute hydrochloric acid. The organic phase was washed with 30 ml of water three times, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain compound 4b.

[0213] 1 H NMR (400MHz, CDCl3) δ = 4.21 (t, J = 6.4Hz, 2H), 3.47 (t, J = 6.4Hz, 2H), 2.25-2.13 (m, 2H), 2.07 (s, 3H).

[0214] Step 2

[0215] Compound 3,5-difluoro-2-hydroxy-benzaldehyde (7.5 g, 47.44 mmol) and compound 4b (11.16 g, 61.67 mmol) were dissolved in 100 ml of N,N-dimethylformamide, and potassium carbonate (13.11 g, 94.88 mmol) was added. The mixture was heated to 75°C and stirred at 75°C for 12 hours. The reaction solution was cooled to 25°C, poured into 200 ml of water, and extracted with ethyl acetate three times, each with 100 ml. The organic phases were combined and washed four times with water, each with 100 ml. The organic phases were concentrated, and the crude product was purified by silica gel column (silica gel, 100-200 mesh, petroleum ether:ethyl acetate = 1 / 0-20 / 1) to obtain compound 4c.

[0216] Step 3

[0217] Compound 4c (12 g, 46.47 mmol) was dissolved in 100 mL of tetrahydrofuran and 10 mL of water, cooled to 0°C, and sodium borohydride (2.11 g, 55.77 mmol) was added portionwise. The mixture was then stirred at 25°C for 3 hours. The reaction was quenched with 1 mol / L dilute hydrochloric acid to a pH of approximately 6, and extracted twice with 50 mL of ethyl acetate. The organic phases were combined, washed once with 100 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated to obtain compound 4d.

[0218] Step 4

[0219] Compound 4d (8 g, 30.74 mmol), 5-fluorosalicylaldehyde (5.6 g, 39.96 mmol), and tri-n-butylphosphine (12.44 g, 61.48 mmol) were dissolved in 50 ml of tetrahydrofuran and cooled to 0°C. Azodicarbonyldipiperidine (15.51 g, 61.48 mmol) was then added portionwise and stirred at 25°C under nitrogen for 12 hours. The reaction solution was poured into 50 ml of water and extracted twice with 50 ml of ethyl acetate. The organic phases were combined and concentrated, and the crude product was purified on a silica gel column (silica gel, 100-200 mesh, petroleum ether:ethyl acetate = 1 / 0-20 / 1) to obtain compound 4e.

[0220] Step 5

[0221] Compound 4e (6.5 g, 17.00 mmol) was dissolved in 55 ml of dichloromethane, and m-chloroperbenzoic acid (4.83 g, 23.80 mmol, purity: 85%) was added. The mixture was then heated to 35°C and stirred at 35°C for 12 hours. The reaction solution was cooled to 25°C and filtered. The filtrate was washed twice with 25 ml of water, twice with saturated sodium bicarbonate, and twice with 30 ml of saturated sodium sulfite solution. The organic phase was concentrated, and the crude product was purified on a silica gel column (silica gel, 100-200 mesh, petroleum ether:ethyl acetate = 1:0-20:1) to obtain compound 4f.

[0222] Step 6

[0223] Compound 4f (5 g, 12.55 mmol) was dissolved in 50 mL of methanol and 10 mL of water, and potassium carbonate (5.2 g, 37.66 mmol) was added, followed by stirring at 25°C for 12 hours. The mixture was filtered and the filtrate was concentrated. The crude product was diluted with 20 mL of ethyl acetate, washed with 20 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain compound 4g.

[0224] Step 7

[0225] Compound 4g (1.63 g, 4.97 mmol) was dissolved in 20 ml of tetrahydrofuran and cooled to 0°C. Sodium hydroxide (595.77 mg, 60% purity, 14.90 mmol) was added, followed by stirring at 0°C for 0.5 hour. p-Toluenesulfonyl chloride (946.61 mg, 4.97 mmol) was then added, and the mixture was heated to 50°C and stirred at 50°C for 24 hours. The reaction solution was cooled to 25°C, quenched by the addition of 20 ml of saturated ammonium chloride solution, and extracted once with 20 ml of ethyl acetate. The mixture was concentrated, and the crude product was purified by silica gel column chromatography (silica gel, 100-200 mesh, petroleum ether:ethyl acetate = 1 / 0-20 / 1) to obtain compound 4h.

[0226] Step 8

[0227] Compound 4h (130 mg, 418.99 μmol) was dissolved in 2 mL of acetic acid, and nitric acid (81.24 mg, 837.99 μmol, purity: 65%) was added. The mixture was then heated to 65°C and stirred at 65°C for 1 hour. The reaction solution was cooled to 25°C, poured into 5 mL of water, and extracted twice with 5 mL of ethyl acetate. The organic phases were combined and washed twice with 5 mL of saturated sodium bicarbonate solution, dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated to obtain compound 4i.

[0228] Step 9

[0229] Compound 4i (110 mg, 309.63 μmol) was dissolved in 3 mL of ethyl acetate. Palladium on carbon (50 mg, 10% purity) was added under nitrogen. The atmosphere was replaced with hydrogen three times and stirred at 25°C under a hydrogen atmosphere (15 psi) for 12 hours. The reaction mixture was filtered and the filtrate was concentrated to obtain compound 4j, which was used directly in the next step.

[0230] LC-MS: m / z=326.1[M+H] + .

[0231] Step 10

[0232] Compound 4j (100 mg, 307.43 μmol) and compound BB-1 (103.09 mg, 307.43 μmol) were dissolved in 5 mL of tetrahydrofuran. Triethylamine (31.11 mg, 307.43 μmol) was added, and the mixture was heated to 70°C and stirred at 70°C for 12 hours. The reaction solution was concentrated, and the crude product was purified by thin-layer chromatography (petroleum ether:ethyl acetate = 3:1) to provide compound 4k.

[0233] LC-MS: m / z = 567.1 [M+H] + .

[0234] Step 11

[0235] Compound 4k (80 mg, 141.22 μmol) was dissolved in 3 mL of tetrahydrofuran and 0.5 mL of methanol. Lithium hydroxide monohydrate (35.56 mg, 847.30 μmol) was dissolved in 0.5 mL of water and added dropwise to the reaction solution, which was then stirred at 25°C for 2 hours. The pH was adjusted to 2-3 with 1 mol / L dilute hydrochloric acid, then diluted with 3 mL of water and extracted with ethyl acetate three times (3 mL each time). The organic phases were combined, washed once with 5 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated to obtain compound 4.

[0236] 1H NMR (400MHz, DMSO-d6) δ=14.45(br s,1H),12.03(s,1H),7.47-7.35(m,3H),7.26(br d,J=10.4Hz,1H),7.18(br d,J=8.4Hz,1H),5.09(s,2H),4.37(br s,2H),4.21(br s,2H),2.09(br s,2H); LC-MS: m / z=521.1[M+H] + .

[0237] Biological test data

[0238] Experimental Example 1 Testing of the Activity of the Compounds of the Invention on Human Gonadotropin-Releasing Hormone Receptor

[0239] Experimental purpose: To detect the inhibitory activity of the test compound on gonadotropin-releasing hormone receptor at the cellular level using FLIPR detection technology. Main experimental materials and sources:

[0240] Fluo-4Direct TM Kit - Invitrogen-F10471

[0241] 384-well poly-lysine-coated cell plates - Greiner-781946

[0242] 384-well compound plate - Greiner-781280

[0243] Compound Preparation ECHO (Acoustic Liquid Handling System) - Labcyte

[0244] FLIPR (Fluorescence Imaging Plate Reader) - Molecular Devices

[0245] Experimental steps:

[0246] GnRH / HEK293 (human embryonic kidney 293) cells in logarithmic growth phase were cultured and washed with DPBS (Dulbecco's phosphate-buffered saline). The cells were digested in a 37°C CO2 incubator with 0.05% EDTA (ethylenediaminetetraacetic acid)-trypsin. After 1-2 minutes, the cells were removed and digested with culture medium. The cells were dispersed by repeated pipetting and harvested by centrifugation. The cells were seeded at a density of 20,000 cells per well in a 384-well polylysine-coated plate using 20 μL of culture medium. The plates were incubated overnight at 37°C in a 5% CO2 incubator.

[0247] On the second day, add 20 μL of 2×Fluo-4 Direct to each well. TMBuffer, 5% CO2, incubate at 37°C incubator for 50 minutes, and place the cells at room temperature for 10 minutes. Use ECHO to make a 4-fold 10-point gradient dilution of 0.2mM Leuprolide acetate and transfer 900nL to the compound plate. Add 30μL FLIPR buffered saline solution to the compound plate and centrifuge at 1000rpm for 1min. Run the FLIPR instrument software and follow the set program to add 10μL of experimental buffered saline solution and read the fluorescence signal. Then add 10μL of agonist reference compound, read the fluorescence signal, and calculate the EC 80 , prepare 6×EC 80 concentration of agonist.

[0248] 2 mM test compound and reference compound at appropriate concentrations were diluted 4-fold in a 10-point series using ECHO, and 900 nL was transferred to the compound plate. 30 μL of FLIPR buffered saline was added to the compound plate and centrifuged at 1000 rpm for 1 minute. The FLIPR instrument software was run and, according to the pre-programmed protocol, 10 μL of test and reference compounds were added to the cell plate and the fluorescence signal was read. 10 μL of 6×EC was then added. 80 concentration of agonist to the cell plate and read the fluorescence signal.

[0249] Calculation of the IC of compounds for calcium influx inhibition of gonadotropin-releasing hormone receptors 50 , that is, in cells stably expressing GnRH receptors, intracellular Ca 2+ The drug concentration at which the flow was inhibited by half was used to calculate the IC of the drug using GraphPad Prism 5.0 software. 50 .

[0250] Experimental results:

[0251] The inhibitory activity of the compounds of the present invention on human gonadotropin-releasing hormone receptor was determined by the above test method, and the measured IC 50 See Table 1:

[0252] Table 1 IC inhibition of human gonadotropin-releasing hormone receptor activity by the compounds of the present invention 50

[0253] Compound number IC 50 (nM) Compound 13.2 Compound 29.2 Compound 310.1 Compound 418.0

[0254] Conclusion: The compounds of the present invention have a significant inhibitory effect on human gonadotropin-releasing hormone receptor.

[0255] Experimental Example 2 In vivo efficacy evaluation

[0256] Experimental purpose: To evaluate the efficacy of the compound of the present invention in a mouse endometriosis model.

[0257] Test plan:

[0258] 2.1 Main reagents and consumables

[0259] C57BL6 / J female mice, Vernier calipers (ARZ-1331), 8-0 sutures, and stereomicroscope.

[0260] 2.2 Experimental steps

[0261] The estrous cycle of 8-week-old female C57BL6 / J mice was detected by vaginal smear observation. Since the entire estrous cycle is about 4 days, mice that meet the specific estrous cycle on the day of the experiment were selected for the experiment based on the actual smear results.

[0262] Modeling: The uterine horns of donor mice in estrus were excised, opened longitudinally and the biopsies were cut into 2×2 mm sections. The recipient mice were anesthetized with isoflurane gas, and a 1 cm incision was made along the midline to expose the abdominal cavity. Endometriosis mice: four uterine fragments of the donor were sutured to the peritoneal wall of mice in estrus; sham-operated mice: abdominal fat fragments of similar size were sutured. 8-0 black silk suture was used to suture the transplanted tissue, abdominal muscles and skin. Sham group (sham-operated group): The abdominal cavity of each Sham group was opened to transplant fat fragments, and the rest of the steps were the same.

[0263] Four weeks after modeling, a second laparotomy was performed, and the volume of the ectopic lesion (V1) was measured and calculated using a vernier caliper to confirm successful modeling. Twelve mice with relatively uniform V1 were divided into two groups of six: vehicle control group and drug group.

[0264] Sham group (sham operation group, 6 rats): normal feeding; Vehicle group (solvent control group, 6 rats): vehicle was gavaged once a day for 8 consecutive weeks, and the solvent was 10% DMAC + 10% solutol + 80% normal saline; Compound group (6 rats): 100 mpk was gavaged at 10:00 am every day for 8 consecutive weeks, and the solvent was 10% DMAC + 10% solutol + 80% normal saline.

[0265] Eight weeks after administration, the mice were sampled and the volume of the ectopic foci (V2) was measured and calculated using a vernier caliper to calculate the inhibition rate. The calculation formula is: ectopic foci volume V = π / 6 × length × width × height, where length represents the length of the ectopic foci, width represents the width of the ectopic foci, and height represents the height of the ectopic foci.

[0266]

[0267] Experimental results: The experimental results are shown in Table 2.

[0268] Table 2 Inhibitory results of the compounds of the present invention in the mouse endometriosis model

[0269]

[0270] Experimental conclusion: The compound of the present invention can significantly inhibit the growth of endometriosis lesion volume in mice and has excellent in vivo efficacy.

[0271] Experimental Example 3 Pharmacokinetic Evaluation

[0272] Experimental purpose: To study the pharmacokinetic properties of the compound of the present invention in mice.

[0273] Test plan:

[0274] Each test compound was mixed with DMAC and vortexed for 2 minutes. The DMAC solution of the test compound was then mixed and vortexed for 2 minutes to prepare a 10 mg / mL clear solution. 0.0600 mL of the 10 mg / mL solution was added to 0.300 mL of Solutol and vortexed for 2 minutes. 2.400 mL of normal saline was then added and vortexed for 2 minutes to obtain a 0.2 mg / mL clear solution for the PO group. 0.500 mL of the PO group dosing solution was vortexed for 2 minutes, 0.0500 mL of DMAC was added, mixed, and vortexed for 2 minutes. Then, 0.0500 mL of Solutol was added and vortexed for 2 minutes. Finally, 0.400 mL of normal saline was added and vortexed for 2 minutes to obtain a 0.1 mg / mL clear solution. The IV injection dosing solution was filtered through a microporous filter.

[0275] Four male CD-1 mice were divided into two groups. Group 1 received a single intravenous dose of 0.5 mg / kg in a 10% DMAC / 10% Solutol / 80% saline vehicle (5 mL / kg). Group 2 received a single oral gavage of 2 mg / kg of the test compound in a 10% DMAC / 10% Solutol / 80% saline vehicle (10 mL / kg). Whole blood was collected at 0.033 (IV only), 0.083, 0.25, 0.5, 1, 2, 4, and 12 hours post-dose. Plasma was obtained after centrifugation at 3200 g at 2-8°C for 10 minutes. Plasma concentrations of the test compound were determined by LC / MS / MS, and pharmacokinetic parameters were calculated using Phoenix WinNonlin software.

[0276] Experimental results:

[0277] The test results are shown in Table 3. The meaning of each parameter: IV: intravenous injection; PO: oral administration; C0: initial blood drug concentration; C max : maximum drug concentration in the systemic circulation; T max : Reach C max Time required; T 1 / 2 : half-life; V dss : apparent volume of distribution; Cl: clearance; AUC: 0-last : Area under the drug-time curve.

[0278] Table 3 Pharmacokinetic (PK) test results of compound 1 in plasma

[0279] PK parameters IV (0.5 mg / kg) PO (2 mg / kg) C0 (nmol / L) 13513--C max (nmol / L)--25800T max (h)--0.500T 1 / 2 (h)5.665.03V dss (L / kg)0.159--Cl(mL / min / kg)0.332--AUC 0-last (h*nmol / L)48307142824 Bioavailability (%) --95.3

[0280] “--” means not tested or data not obtained

[0281] Conclusion: The compound of the present invention has high exposure in plasma, low clearance rate, long half-life, high oral bioavailability, and exhibits excellent pharmacokinetic properties.

Claims

1. A compound represented by formula (II) or a pharmaceutically acceptable salt thereof, wherein, L 1 and L 2 are each independently selected from -(CH 2 ) n -; L 3 and L 4 are each independently selected from -CH 2 -, -CH=CH-, -O- and -S-; R 1 and R 2 are each independently selected from H, OH, F, Cl, C 1-3 alkyl, and C 1-3 alkoxy, said C 1-3 alkyl and C 1-3 alkoxy are each independently optionally substituted by 1, 2 or 3 R a substituents; Alternatively, R 1 and R 2 together with the atoms to which they are commonly attached form a C 3-6 cycloalkyl or 4- to 6-membered heterocycloalkyl, the C 3-6 cycloalkyl or 4- to 6-membered heterocycloalkyl being optionally substituted by 1, 2 or 3 R b substituents; R 3 、R 4 、R 5 、R 6 、R 7 、R 8 and R 9 are each independently selected from H, F, Cl, Br, I, OH, NH 2 , CN, C 1-3 alkyl, C 1-3 alkoxy and C 3-6 cycloalkyl, wherein the C 1-3 alkyl, C 1-3 alkoxy and C 3-6 cycloalkyl are each independently optionally substituted with 1, 2 or 3 R c substituents; n is selected from 0, 1, and 2; Each R a 、R b and R c are each independently selected from H, F, Cl, Br, I, NH 2 and OH.

2. The compound or a pharmaceutically acceptable salt thereof according to claim 1, wherein, R 1 and R 2 are each independently selected from H, OH, F, and CH 3 , where the CH 3 is optionally substituted with 1, 2, or 3 F atoms.

3. The compound or a pharmaceutically acceptable salt thereof according to claim 2, wherein, R 1 and R 2 are each independently selected from H.

4. The compound or a pharmaceutically acceptable salt thereof according to claim 1, wherein, R 1 and R 2 together with the atoms to which it is commonly attached form cyclopropyl, cyclobutyl, cyclopentyl, oxetanyl or azetidinyl, which cyclopropyl, cyclobutyl, cyclopentyl, oxetanyl or azetidinyl is optionally substituted with 1, 2 or 3 F.

5. The compound or a pharmaceutically acceptable salt thereof according to claim 4, wherein, R 1 and R 2 together with the atoms to which it is jointly attached form 6. The compound or a pharmaceutically acceptable salt thereof according to claim 1, wherein, R 3 、R 4 、R 5 and R 6 are each independently selected from H and F.

7. The compound or a pharmaceutically acceptable salt thereof according to claim 6, wherein, Structural unit 8. The compound or a pharmaceutically acceptable salt thereof according to claim 1, wherein, R 7 、R 8 and R 9 are each independently selected from H and F.

9. The compound or a pharmaceutically acceptable salt thereof according to claim 8, wherein, Structural unit 10. The compound or a pharmaceutically acceptable salt thereof according to claim 1, wherein, L 3 and L 4 are each independently selected from O.

11. The compound or a pharmaceutically acceptable salt thereof according to claim 1, wherein, n is selected from 1 and 2.

12. The compound or a pharmaceutically acceptable salt thereof according to claim 1, wherein, Structural unit 13. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 12, wherein the compound is selected from, wherein, L 1 and L 2 、R 1 and R 2 、R 3 and R 4 、R 5 and R 6 are respectively defined as any one of claims 1 to 12.

14. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 12, wherein the compound is selected from, wherein, L 1 and L 2 are each independently selected from -(CH 2 ) n -; L 3 and L 4 are each independently selected from -CH 2 -, -CH=CH-, -O- and -S-; R 1 and R 2 are each independently selected from H, OH, F, Cl, C 1-3 alkyl, and C 1-3 alkoxy, wherein the C 1-3 alkyl and C 1-3 alkoxy are optionally substituted with 1, 2 or 3 R a substituents; Alternatively, R 1 and R 2 together with the atom(s) to which it is attached form a 3-6 cycloalkyl or 4-6 membered hetero cycloalkyl, wherein the 3-6 cycloalkyl or 4-6 membered hetero cycloalkyl is optionally substituted with 1, 2 or 3 R b substituents; R 3 、R 4 、R 5 and R 6 are each independently selected from H, F, Cl, Br, and I; n is selected from 0, 1, and 2; Each R a and R b are each independently selected from H, F, Cl, Br, I; The "4-6 membered heteroalkyl" contains 1, 2, or 3 heteroatoms selected from N, NH, O, and S.

15. The following compound or a pharmaceutically acceptable salt thereof, 16. Use of the compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 15 in the preparation of a drug related to a GnRH receptor antagonist.

17. The use according to claim 16, wherein, The drug related to the GnRH receptor antagonist is a drug for preventing and / or treating endometriosis and / or uterine fibroids related diseases.

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