ROR gamma regulator and application thereof

Abstract

The invention discloses a compound with a structure as shown in a formula (I) or a stereoisomer, a tautomer or a pharmaceutically acceptable salt or a solvate or a prodrug thereof: # imgabs0 #. The compound with the formula (I) has an obvious inhibition effect on ROR gamma t, and the ROR gamma t has a very important effect in inflammatory, metabolic and autoimmune diseases; the ROR [gamma] t is inhibited, so that the diseases can be relieved or effectively treated.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to retinoic acid-related orphan receptor gamma (RORγ) modulators, preparation methods thereof, pharmaceutical compositions containing these modulators, and their medical uses in treating RORγ-mediated inflammatory, metabolic and autoimmune diseases. Background Art

[0002] Retinoic acid receptor-related orphan receptors (RORs) are a class of ligand-dependent transcription factors that play an important role in a series of physiological and pathological processes such as reproductive development, physiological rhythm regulation, metabolic disorders, inflammation, and immune system regulation. RORs are a member of the nuclear receptor superfamily, including RORα, RORβ, and RORγ. RORα is mainly distributed in the liver, skeletal muscle, skin, lung, adipose tissue, kidney, thymus, brain, and blood, and is related to physiological and pathological processes such as hepatic gluconeogenesis, lipid metabolism, and atherosclerosis. RORβ is mainly distributed in the central nervous system, including the brain, retina, and pineal gland, and is mainly related to the processing of sensitive information in the spinal cord, thalamus, and cerebellar cortex. RORγ is highly expressed in the thymus, and is also distributed in the kidney, liver, heart, skeletal muscle, adipose tissue, testis, prostate, and pancreas, and is related to autoimmune diseases such as rheumatoid arthritis, psoriasis, and multiple sclerosis.

[0003] RORγ includes two subtypes, RORγ1 and RORγ2 (RORγt). RORγ1 is expressed in a variety of tissues including the thymus, muscle, kidney and liver; RORγt is uniquely expressed in immune system cells and plays a key role in thymic generation, the development of several secondary lymphoid tissues and the differentiation of the Th17 lineage. Studies have shown that RORγt is a key regulator of Th17 cell differentiation. Th17 cells are a subtype of T helper cells that produce IL-17 and other proinflammatory cytokines. Th17 cells play a key role in several mouse autoimmune disease models, including encephalomyelitis (EAE) and collagen-induced arthritis (CIA). In addition, studies have shown that Th17 cells or their products are associated with the pathology of a variety of human inflammatory and autoimmune diseases, including multiple sclerosis, rheumatoid arthritis, psoriasis, Crohn's disease, and asthma. The main cause of the onset of autoimmune diseases is intolerance to self-antigens and the development of auto-invasive effector T cells that infiltrate tissues. Th17 cells are one of the important driving factors in the inflammatory process of tissue-specific autoimmunity. During the development of the disease, Th17 cells are activated and are responsible for recruiting other inflammatory cells (neutrophils), thereby mediating lesions in target tissues.

[0004] RORγt has been reported as a key regulator of Th17 cell differentiation, and Th17 cells have recently been found to be a subset of T helper cells that preferentially produce the cytokines IL-17A, IL-17F, IL-21, and IL-22. RORγt induces transcription of genes encoding IL-17A and IL-17F in naive CD4+ T helper cells. Mice deficient in RORγt show very few Th17 cells. Inhibition and deletion of RORγt improve EAE.

[0005] In asthmatic patients, RORγt and IL-17A expression levels have been shown to be increased in saliva, lungs, bronchoalveolar lavage (BAL) fluid, and peripheral blood, and levels are directly correlated with disease severity. In addition to IL-17A, recent studies have shown that another cytokine of the IL-17 family, IL-17F, may play an important role in allergic airway inflammation and therefore has an important influence in airway diseases such as asthma. Overexpression of the IL-17F gene in the airways of mice is associated with airway neutrophilia, cytokine induction, increased airway hyperresponsiveness, and excessive mucus secretion.

[0006] In view of the role that RORγ plays in the pathogenesis of disease, it is desirable to prepare compounds that modulate RORγ activity and are therefore useful in treating RORγ-mediated inflammatory, metabolic, and autoimmune diseases, such as the respiratory diseases asthma, chronic obstructive pulmonary disease (COPD) and bronchitis, allergic diseases including allergic rhinitis and atopic dermatitis, cystic fibrosis, and lung allograft rejection. Summary of the invention

[0007] 1. Technical problems to be solved:

[0008] In view of the role that RORγ plays in the pathogenesis of disease, it is desirable to prepare compounds that modulate RORγ activity and are therefore useful in treating RORγ-mediated inflammatory, metabolic and autoimmune diseases.

[0009] In view of the above technical problems, the present invention provides retinoic acid-related orphan receptor γ (RORγ) modulators, preparation methods thereof, pharmaceutical compositions containing these modulators and their medical uses in treating RORγ-mediated inflammatory, metabolic and autoimmune diseases.

[0010] 2. Technical solution:

[0011] A compound having a structure as shown in formula (I) or a stereoisomer, a tautomer or a pharmaceutically acceptable salt or a solvate or a prodrug thereof:

[0012]

[0013] in,

[0014] R1 is selected from hydrogen, halogen, C1-C3 alkyl, C1-C3 haloalkyl, alkoxy;

[0015] X is selected from N or CR6, R6 is selected from halogen, C1-C3 alkyl, C1-C3 haloalkyl, alkoxy;

[0016] Ring A is a 5-10 membered heteroaryl or a 6-10 membered aromatic group; R2 is selected from a C1-C6 alkyl group, a C3-C8 cycloalkyl group, a 3-8 membered heterocycloalkyl group, an aryl group, a heteroaryl group, and a cyano group;

[0017] R 3、 R4 is independently selected from hydrogen, halogen, C1-C6 alkyl and C1-C6 haloalkyl, or R 3、 R4 merges into a ring;

[0018] Ring B is a 5-10 membered heterocycloalkyl, a 6-10 membered aryl or a 5-10 membered heteroaryl, and R5 is selected from halogen, hydroxy, amino, C3-C6 cycloalkyl or C1-C6 alkyl.

[0019] Furthermore, the R1 is selected from halogen, methyl, ethyl, propyl, isopropyl, n-butyl, tert-butyl, -CF3.

[0020] Furthermore, X is selected from N or CR6, and R6 is Cl.

[0021] Furthermore, the ring A is one of a benzene ring, a pyrimidine ring and a thiazole ring.

[0022] Furthermore, the R2 is selected from C1-C6 alkyl and cyano.

[0023] Furthermore, R3 and R4 are independently selected from hydrogen, halogen, methyl, ethyl, propyl, isopropyl, n-butyl and tert-butyl.

[0024] Furthermore, the ring B comprises: “*” is the connection position with N.

[0025] Further, R5 is a C1-C6 alkyl group, including methyl, ethyl, propyl, isopropyl, n-butyl and tert-butyl.

[0026] Further, the compound represented by formula (I) includes:

[0027]

[0028] Furthermore, use of the compound or stereoisomer, tautomer or pharmaceutically acceptable salt or solvate or prodrug thereof in the preparation of a drug for treating RORγ-mediated diseases.

[0029] Furthermore, the disease is an inflammatory, metabolic or autoimmune disease.

[0030] Further, the inflammatory, metabolic or autoimmune disease includes asthma, chronic obstructive pulmonary disease, bronchitis, allergic rhinitis, atopic dermatitis, cystic fibrosis, lung allograft rejection, multiple sclerosis, rheumatoid arthritis, juvenile rheumatoid arthritis, osteoarthritis, ankylosing spondylitis, systemic lupus erythematosus, psoriasis, Hashimoto's disease, pancreatitis, autoimmune diabetes, autoimmune eye disease, ulcerative colitis, Crohn's disease, inflammatory bowel disease, inflammatory bowel syndrome, Sjogren's syndrome, optic neuritis, type I diabetes, neuromyelitis optica, myasthenia gravis, uveitis, Guillain-Barré syndrome, psoriatic arthritis, Grave's disease or scleritis.

[0031] Furthermore, the disease is asthma, rheumatoid arthritis, psoriasis, ulcerative colitis or Crohn's disease.

[0032] The terms used in the specification and claims of the present invention have the following meanings.

[0033] "Alkyl" refers to a saturated aliphatic hydrocarbon group, including straight or branched chain groups of 1 to 20 carbon atoms. 1-6 Alkyl refers to a medium-sized alkyl group containing 1 to 6 carbon atoms, such as methyl, ethyl, propyl, 2-propyl, n-butyl, isobutyl, tert-butyl, pentyl, etc. Preferred are lower alkyl groups containing 1 to 4 carbon atoms, such as methyl, ethyl, propyl, 2-propyl, n-butyl, isobutyl or tert-butyl, etc. The alkyl group may be substituted or unsubstituted. When substituted, preferred groups are: halogen, C2-C6 alkenyl, C6-C 10 Aryl, C5-C 10 Heteroaryl, halogenated C1-C6 alkyl, 4- to 8-membered heteroalicyclic group, hydroxyl, C1-C6 alkoxy, C6-C 10 Aryloxy.

[0034] "Alkylamino" refers to a group in which one or two hydrogen atoms in an amino group are replaced by an alkyl group. It includes amino groups substituted by straight-chain, branched-chain or cyclic alkyl groups, such as methylamino, dimethylamino, ethylamino, n-propylamino, isopropylamino, n-butylamino, isobutylamino, tert-butylamino, cyclopropylamino, cyclobutylamino, pentylamino, etc. Preferred are amino groups substituted by lower straight-chain, branched-chain or cyclic alkyl groups containing 1 to 4 carbon atoms.

[0035] "Cycloalkyl" refers to a 3-8-membered all-carbon monocyclic ring, an all-carbon 5-membered / 6-membered or 6-membered / 6-membered fused ring or a polycyclic fused ring ("fused" ring means that each ring in the system shares a pair of adjacent carbon atoms with other rings in the system) group, wherein one or more rings have a completely connected π electron system, and examples of cycloalkyl (not limited to) are cyclopropane, cyclobutane, cyclopentane, cyclopentene, cyclohexane, adamantane, cyclohexadiene, cycloheptane or cycloheptatriene. Cycloalkyl is substituted and unsubstituted. When substituted, the substituents are preferably one or more groups each selected from the group consisting of hydrogen, hydroxy, mercapto, oxo, lower alkyl, lower alkoxy, lower cycloalkyl, lower heterocycloalkyl, lower haloalkoxy, alkylthio, halogen, lower haloalkyl, lower hydroxyalkyl, lower cycloalkylalkylene, lower heterocycloalkylalkylene, aryl, heteroaryl, alkoxycarbonyl, amino, alkylamino, alkylsulfonyl, arylsulfonyl, alkylaminosulfonyl, arylaminosulfonyl, alkylsulfonylamino, arylsulfonylamino, alkylaminocarbonyl, arylaminocarbonyl, alkylcarbonylamino, arylcarbonylamino.

[0036] "Aryl" refers to an all-carbon monocyclic or fused polycyclic radical of 6 to 14 carbon atoms, having a completely conjugated π electron system. "Aryl" includes:

[0037] Six-membered carbon aromatic ring, such as benzene;

[0038] Bicyclic rings, at least one of which is a carbon aromatic ring, such as naphthalene, indene or 1,2,3,4-tetrahydroquinoline; and

[0039] A tricyclic ring, at least one of which is a carbon aromatic ring, such as fluorene.

[0040] For example, aryl includes a six-membered carbon aromatic ring and a six-membered heterocyclic ring, which heterocyclic ring contains one or more heteroatoms selected from nitrogen, oxygen and sulfur, provided that the point of attachment is on the carbon aromatic ring. However, aryl does not contain or overlap with the heterocyclic aromatic groups defined below in any way. Therefore, it is defined here that if one or more carbon aromatic rings are fused with a heteroaromatic ring, the resulting ring system is a heteroaryl group, not an aryl group. Non-limiting examples of aryl groups include phenyl and naphthyl. Aryl groups can be substituted or unsubstituted. When substituted, preferred groups are: hydrogen, hydroxy, nitro, cyano, oxo, lower alkyl, lower alkoxy, lower cycloalkyl, lower heterocycloalkyl, lower haloalkoxy, alkylthio, halogen, lower haloalkyl, lower hydroxyalkyl, lower cycloalkylalkylene, lower heterocycloalkylalkylene, aryl, heteroaryl, alkoxycarbonyl, amino, alkylamino, alkylsulfonyl, arylsulfonyl, alkylaminosulfonyl, arylaminosulfonyl, alkylsulfonylamino, arylsulfonylamino, alkylaminocarbonyl, arylaminocarbonyl, alkylcarbonylamino, arylcarbonylamino.

[0041] "Heteroaryl" means a monocyclic or fused ring radical of 5 to 14 ring atoms, containing one, two, three or four ring heteroatoms selected from N, O or S, the remaining ring atoms being C, and having a completely conjugated π electron system. Heteroaryl refers to:

[0042] 5-8 membered monocyclic aromatic hydrocarbons containing one or more heteroatoms selected from N, O and S, such as 1-4 heteroatoms, in some embodiments, 1-3 heteroatoms, and the other atoms in the ring are carbon atoms;

[0043] 8-12 membered bicyclic aromatic hydrocarbons containing one or more heteroatoms selected from N, O and S, such as 1-4 heteroatoms, in some embodiments, 1-3 heteroatoms, the other atoms in the ring being carbon atoms; wherein at least one ring is aromatic; and

[0044] 11-14 membered tricyclic aromatic hydrocarbons, containing one or more heteroatoms selected from N, O and S, such as 1-4 heteroatoms, in some embodiments, 1-3 heteroatoms, and the other atoms on the ring are carbon atoms; wherein at least one ring is an aromatic ring.

[0045] For example, the heteroaryl group includes a 5-6 membered heteroaromatic ring and a 5-6 membered cycloalkyl group. For such bicyclic heteroaromatic groups, only one of which contains one or more heteroatoms, the point of attachment is on the heteroaromatic ring.

[0046] When the total number of sulfur atoms and oxygen atoms in the heteroaryl group exceeds 1, these heteroatoms are not adjacent to each other. In some embodiments, the total number of sulfur atoms and oxygen atoms in the heteroaryl group does not exceed 2. In some embodiments, the total number of sulfur atoms and oxygen atoms in the heteroaryl group does not exceed 1.

[0047] Examples of heteroaryl groups include, but are not limited to, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, triazole, pyrimidine, pyridine, pyridone, imididine, pyrazine, pyridazine, indole, azaindole, benzimidazole, benzotriazole, indoline, indolone, quinoline, isoquinoline, quinazoline, thienopyridine, thienopyrimidine, etc. Preferred embodiments of such groups are benzene rings, pyridine, pyrimidine, pyridazine, pyrazine, quinoline, isoquinoline, pyrrole, pyrazole, imidazole, thiophene, thiazole, furan or oxazole. One or all of the hydrogen atoms in the heteroaryl group may be substituted by hydrogen, hydroxy, nitro, cyano, oxo, lower alkyl, lower alkoxy, lower cycloalkyl, lower heterocycloalkyl, lower haloalkoxy, alkylthio, halogen, lower haloalkyl, lower hydroxyalkyl, lower cycloalkylalkylene, lower heterocycloalkylalkylene, aryl, heteroaryl, alkoxycarbonyl, amino, alkylamino, alkylsulfonyl, arylsulfonyl, alkylaminosulfonyl, arylaminosulfonyl, alkylsulfonylamino, arylsulfonylamino, alkylaminocarbonyl, arylaminocarbonyl, alkylcarbonylamino, and arylcarbonylamino.

[0048] "Heterocycloalkyl" means a monovalent saturated cyclic group consisting of one or more rings, preferably 1 to 2 rings (including spirocyclic systems), each ring 3 to 8 atoms, which is combined with one or more ring heteroatoms (selected from N, O or S (O) 0-2 ), and it may be optionally independently substituted by one or more, preferably 1 or 2 substituents selected from the group consisting of hydrogen, hydroxyl, mercapto, oxo, lower alkyl, lower alkoxy, lower cycloalkyl, lower heterocycloalkyl, lower haloalkoxy, alkylthio, halogen, lower haloalkyl, lower hydroxyalkyl, lower cycloalkyl alkylene, lower heterocycloalkyl alkylene, aryl, heteroaryl, alkoxycarbonyl, amino, alkylamino, alkylsulfonyl, arylsulfonyl, alkylaminosulfonyl, arylaminosulfonyl, alkylsulfonylamino, arylsulfonylamino, alkylaminocarbonyl, arylaminocarbonyl, alkylcarbonylamino, arylcarbonylamino. Unless otherwise indicated.

[0049] Examples of heterocycloalkyl groups include, but are not limited to, oxirane, aziridine, pyridine, morpholin-3-one, thiomorpholine 1,1-dioxide, morpholinyl, piperazinyl, piperidinyl, azetidinyl, pyrrolidinyl, hexahydroazepine, oxetanyl, tetrahydrofuranyl, tetrahydrothiophenyl, oxazolidinyl, thiazolidinyl, isoxazolidinyl, tetrahydro-2H-pyranyl, thiomorpholinyl, quinuclidinyl and imidazolinyl, preferably

[0050] W is selected from O, S or NR 12 , each group is as described above, examples can also be bicyclic, such as, for example, 3,8-diaza-bicyclo[3.2.1]octane, 2,5-diazabicyclo[2.2.2]octane or octahydro-pyrazino[2,1-c][1,4]oxazine; preferably oxirane, oxetane, tetrahydrofuran, tetrahydro-2H-pyran, aziridine, azetidine, pyrrolidine, piperidine, morpholine, pyridine, morpholine-3-one or thiomorpholine 1,1-dioxide; its heterocycloalkyl (and derivatives) include its ionic form.

[0051] "Alkoxy" refers to -O-(unsubstituted alkyl) and -O-(unsubstituted cycloalkyl). Representative examples include, but are not limited to, methoxy, ethoxy, propoxy, butoxy, cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy, and the like.

[0052] "Aryloxy" refers to -O-aryl and -O-heteroaryl. Representative examples include, but are not limited to, phenoxy, pyridyloxy, furanyloxy, thienyloxy, pyrimidyloxy, pyrazinyloxy, and the like and their derivatives.

[0053] "Arylalkylene" means an alkyl group, preferably a lower alkyl group as defined above, substituted with an aryl group as defined above, for example -CH2phenyl, -(CH2)2phenyl, -(CH2)3phenyl, CH3CH(CH3)CH2phenyl and derivatives thereof.

[0054] "Heteroarylalkylene" means an alkyl group, preferably a lower alkyl group as defined above, which is substituted with a heteroaryl group as defined above, for example -CH2 pyridyl, -(CH2)2 pyrimidinyl, -(CH2)3 imidazolyl, and the like, and derivatives thereof.

[0055] "Oxo" means a =0 group.

[0056] "Hydroxy" means an -OH group.

[0057] "Mercapto" means a -SH group.

[0058] "Halogen" means fluorine, chlorine, bromine or iodine, preferably fluorine or chlorine.

[0059] "Haloalkyl" means an alkyl group substituted by halogen, preferably a lower alkyl group as defined above, which is substituted by one or more halogen atoms which may be the same or different, for example -CH2Cl, -CF3, -CCl3, -CH2CF3, -CH2CCl3, etc.

[0060] "Cyano" means a -CN group.

[0061] "Amino" means a -NH2 group.

[0062] "Nitro" means a -NO2 group.

[0063] "Tetrahydro-2H-pyran" means

[0064] "Alkylsulfonyl" means -S(O2)C 1-6 Alkyl, wherein alkyl is as defined above.

[0065] "Optionally substituted" includes both the case where one or more substituents replace the group and the case where the group is unsubstituted. For example, an optionally substituted alkyl group includes an unsubstituted alkyl group and an alkyl group substituted by one or more substituents.

[0066] The term "optionally" means that the subsequently described event or circumstance may or may not occur, and that the description includes that the event or circumstance may or may not occur, and that the description includes both situations where the event or circumstance occurs and where it does not occur.

[0067] In some embodiments, "substituted with one or more groups" means that one, two, three or four hydrogen atoms in the specified atom or group are replaced by the same or different groups selected from the specified range of groups, respectively.

[0068] Wavy lines indicate junction sites;

[0069] "Pharmaceutically acceptable salts" refers to those salts that retain the biological effectiveness and properties of the parent compound. Such salts include:

[0070] (1) Acid salts are obtained by reacting the free base of the parent compound with an inorganic acid or an organic acid. The inorganic acids include hydrochloric acid, hydrobromic acid, nitric acid, phosphoric acid, metaphosphoric acid, sulfuric acid, sulfurous acid and perchloric acid. The organic acids include acetic acid, propionic acid, acrylic acid, oxalic acid, (D) or (L) malic acid, fumaric acid, maleic acid, hydroxybenzoic acid, γ-hydroxybutyric acid, methoxybenzoic acid, phthalic acid, methanesulfonic acid, ethanesulfonic acid, naphthalene-1-sulfonic acid, naphthalene-2-sulfonic acid, p-toluenesulfonic acid, salicylic acid, tartaric acid, citric acid, lactic acid, mandelic acid, succinic acid or malonic acid.

[0071] (2) Salts formed by the replacement of acidic protons in the parent compound by metal ions or by coordination with organic bases. Examples of metals include alkali metal ions, alkaline earth metal ions or aluminum ions; and organic bases include ethanolamine, diethanolamine, triethanolamine, tromethamine, N-methylglucamine, etc.

[0072] "Pharmaceutical composition" refers to a combination of one or more of the compounds of the present invention or their pharmaceutically acceptable salts, solvates, hydrates or prodrugs with one or more pharmaceutically acceptable excipients. Among them, "excipients" are usually other chemical components other than the compounds of the present invention, such as pharmaceutically acceptable pharmaceutical carriers, or other compounds with pharmaceutical effects. The purpose of the pharmaceutical composition can be to facilitate the process of administration to animals, or it can be a drug synergistic effect.

[0073] "Pharmaceutical carrier" refers to an inactive ingredient in a pharmaceutical composition that does not cause significant irritation to an organism and does not interfere with the biological activity and properties of the administered compound, such as but not limited to: calcium carbonate, calcium phosphate, various sugars (such as lactose, mannitol, etc.), starch, cyclodextrin, magnesium stearate, cellulose, magnesium carbonate, acrylic acid polymers or methacrylic acid polymers, gelatin, water, polyethylene glycol, propylene glycol, ethylene glycol, castor oil or hydrogenated castor oil or polyethoxylated hydrogenated castor oil, sesame oil, corn oil, peanut oil, etc.

[0074] The aforementioned pharmaceutical composition, in addition to pharmaceutically acceptable carriers, may also include adjuvants commonly used in pharmacy, such as antibacterial agents, antifungal agents, antimicrobial agents, preservatives, colorants, solubilizers, thickeners, surfactants, chelating agents, proteins, amino acids, fats, carbohydrates, vitamins, minerals, trace elements, sweeteners, pigments, flavors, or combinations thereof.

[0075] 3. Beneficial effects:

[0076] The compound of formula (I) described in the present invention has a significant inhibitory effect on RORγt, and RORγt plays a very important role in inflammatory, metabolic and autoimmune diseases. Inhibiting RORγt will alleviate or effectively treat these diseases. DETAILED DESCRIPTION

[0077] The present invention is described in detail below.

[0078] Example 1: Preparation of 6-(3-((4-(5-(tert-butyl)-1,2,4-oxadiazol-3-yl)-3-methylpiperazin-1-yl)methyl)-5-chloro-2-(trifluoromethyl)phenyl)-2-methyl-7,8-dihydropyrido[4,3-d]pyrimidin-5(6H)-one EXP 1

[0079]

[0080] 1) Preparation of Compound 3

[0081] Compound 1 (2.24 g, 10 mmol, 1 eq), Na2CO3 (5.30 g, 50 mmol, 5 eq), compound 2 (3.70 g, 12 mmol, 1.2 eq), and 80 mL of toluene were added to a 100 ml single-mouth bottle, replaced with nitrogen three times, heated to 100 ° C, reacted for 3 hours, and the reaction was completed by TLC detection. After the reaction was completed, the toluene was concentrated to remove the toluene, EA (200 ml) * 3 and ice water (30 ml) were added, the organic phase was separated, washed with saturated brine (50 ml), the organic phase was dried with magnesium sulfate, and concentrated to obtain a crude product. Prep-HPLC preparation, obtained off-white solid compound 3 3.96 g, yield 80%. LC-MS [M+1] + :496.50; 1 H NMR (400MHz, CDCl3): 7.62(s,1H),7.33(s,1H),3.62(d,2H),2.61-2.73(m,7H),1.28-1.35(m,9H),1.22(m,3H).

[0082] 2) EXP 1 Preparation

[0083] Compound 3 (2.48 g, 5 mmol, 1 eq), compound 4 (0.98 g, 6.0 mmol, 1.2 eq), Pd(dppf)Cl2 (570 mg, 0.78 mmol, 0.16 eq), potassium phosphate (4.97 g, 23.4 mmol, 4.7 eq) were added to a 100 ml single-mouth bottle, dioxane (50 ml), water (2 ml), replaced with nitrogen 3 times, heated to 100 ° C, reacted for 3 hours, and the reaction was completed by TLC detection. The reaction solution was cooled to room temperature, EA (300 ml) * 3, water (30 ml) were added, the organic phase was separated, washed with saturated brine (50 ml), the organic phase was dried with magnesium sulfate, spin-dried, and prepared by Prep-HPLC to obtain an off-white solid EXP 1 1.74 g, with a yield of 60%. LC-MS [M+1] + : 577.2; 1 H NMR(400MHz, CDCl3):8.66(s,1H),7.72(s,1H),7.13(s,1H),3.96(m,2H),3.60(d, 2H),3.10(m,2H),2.60-2.74(m,7H),2.45(s,3H),1.31-1.35(m,9H),1.22(s,3H).

[0084] Example 2: Preparation of 2-(3-((4-(5-(tert-butyl)-1,2,4-oxadiazol-3-yl)-3-methylpiperazin-1-yl)methyl)-5-chloro-2-(trifluoromethyl)phenyl)-1-oxo-1,2,3,4-tetrahydroisoquinoline-6-carbonitrile EXP 2

[0085]

[0086] Referring to the method of Example 1, 1.25 g of off-white solid compound EXP 2 was prepared with a yield of 68%. LC-MS [M+1] + :588.20. 1 H NMR(400MHz, CDCl3):8.21(s,1H),8.03(s,1H),7.78-7.85(m,2H),7.19(s,1H),3.60 (d,2H),3.56(m,2H),3.15(m,2H),2.54-2.74(m,7H),1.33-1.38(m,9H),1.24(s,3H).

[0087] Example 3: Preparation of 2-(5-((4-(5-(tert-butyl)-1,2,4-oxadiazol-3-yl)-3-methylpiperazin-1-yl)methyl)-4-(trifluoromethyl)pyridin-3-yl)-6-methyl-3,4-dihydroisoquinolin-1(2H)-one EXP 3

[0088]

[0089] Referring to the method of Example 1, 1.47 g of off-white solid compound EXP 3 was prepared with a yield of 88%. LC-MS [M+1] + :543.20. 1 H NMR(400MHz, CDCl3):8.51(s,1H),8.43(s,1H),8.03(d,1H),7.26-7.29(m,2H),3.63 (m,2H),3.54(q,2H),3.15(m,2H),2.74-2.84(m,7H),2.15(t,3H),1.33-1.38(m,9H).

[0090] Example 4: Preparation of 5-(3-((4-(5-(tert-butyl)-1,2,4-oxadiazol-3-yl)-3-methylpiperazin-1-yl)methyl)-5-chloro-2-(trifluoromethyl)phenyl)-2-methyl-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one EXP 4

[0091]

[0092] Referring to the method of Example 1, 1.87 g of off-white solid compound EXP 4 was prepared with a yield of 78%. LC-MS [M+1] + :584.20. 1 H-NMR(400MHz, CDCl3):7.78(m,1H),7.23(s,1H),3.93(t,2H),3.65(d,2H),3.1 4(m,2H),2.78-2.84(m,4H),2.75(t,3H),2.48-2.64(m,3H),1.33-1.37(m,9H).

[0093] Example 5: Preparation of 6-(5-chloro-3-((4-(4,4-dimethyl-4,5-dihydrooxazol-2-yl)-3-methylpiperazin-1-yl)methyl)-2-methylphenyl)-2-methyl-7,8-dihydropyrido[4,3-d]pyrimidin-5(6H)-one EXP 5

[0094]

[0095] Referring to the method of Example 1, 1.68 g of off-white solid compound EXP 5 was prepared with a yield of 72%. LC-MS [M+1] + :498.20. 1 H-NMR(400MHz, CDCl3):8.68(m,1H),7.76(s,1H),7.24(s,1H),3.95(t,2H),3.85(d,2H),3.64(m,2H),3.44-3.52(m,2H ),3.13(m,2H),2.79-2.84(m,2H),2.73(s,1H),2.48-2.60(m,2H),2.35(t,3H),2.15(m,3H),1.33(m,3H),1.03(m,6H).

[0096] Example 6: Preparation of 6-(3-((4-(5-(tert-butyl)isoxazol-3-yl)-3-methylpiperazin-1-yl)methyl)-5-chloro-2-(trifluoromethyl)phenyl)-2-methyl-7,8-dihydropyrido[4,3-d]pyrimidin-5(6H)-one EXP 6

[0097]

[0098] Referring to the method of Example 1, 1.28 g of off-white solid compound EXP 6 was prepared with a yield of 52%. LC-MS [M+1] + :578.20. 1 H-NMR(400MHz, CDCl3):8.67(s,1H),7.78(s,1H),7.24(s,1H),6.65(s,1H),3.95(m,2H),3.65(d ,2H),3.14(m,2H),2.82-2.84(m,4H),2.73(s,1H),2.41-2.60(m,5H),1.35(m,9H),1.23(m,3H).

[0099] Example 7: Preparation of 6-(3-((4-(5-(tert-butyl)thiazol-2-yl)-3-methylpiperazin-1-yl)methyl)-5-chloro-2-(trifluoromethyl)phenyl)-2-methyl-7,8-dihydropyrido[4,3-d]pyrimidin-5(6H)-one EXP 7

[0100]

[0101] Referring to the method of Example 1, 0.28 g of off-white solid compound EXP 7 was prepared with a yield of 62%. LC-MS [M+1] + :594.10.1 H-NMR(400MHz, CDCl3):8.71(s,1H),7.76(s,1H),7.27(s,1H),6.85(s,1H),3.96(m,2H),3.64(d ,2H),3.10(m,2H),2.80-2.84(m,4H),2.67(s,1H),2.40-2.54(m,5H),1.36(m,9H),1.21(m,3H).

[0102] Example 8: Preparation of 6-(3-((4-(5-(tert-butyl)-1,2,4-oxadiazol-3-yl)-3,3-dimethylpiperazin-1-yl)methyl)-5-chloro-2-(trifluoromethyl)phenyl)-2-methyl-7,8-dihydropyrido[4,3-d]pyrimidin-5(6H)-one EXP 8

[0103]

[0104] Referring to the method of Example 1, 0.68 g of off-white solid compound EXP 8 was prepared with a yield of 32%. LC-MS [M+1] + :593.23. 1 H-NMR(400MHz, CDCl3):8.70(s,1H),7.79(s,1H),7.25(s,1H),3.96(m,2H),3.65(m,2H),3 .16(d,2H),3.10(m,2H),2.70(m,2H),2.47(s,2H),2.40(m,3H),1.37(m,6H),1.31(m,9H).

[0105] Example 9 RORγt inhibitor luciferase reporter gene experiment

[0106] Experimental materials and instruments:

[0107]

[0108]

[0109] SR1001 is an inverse agonist of RORγt and is used as a positive reference. Its structure is:

[0110]

[0111] Experimental steps:

[0112] 1. On the first day, cells were plated. 293T adherent cells were digested with 1 mL of trypsin for about 5 minutes. The digested cells were aspirated with a pipette and transferred to a 15 mL centrifuge tube. Centrifuged at 1000 rpm for 5 minutes. The old culture medium was discarded, and the cells were resuspended with fresh culture medium and diluted to the desired density.

[0113] 2. Count cells. Prepare cell suspension at a cell density of 15,000 cells / well. Seed the plate with 100 μL of cells per well. To prevent edge effects, only the middle 60 wells of the 96-well cell culture plate are seeded, and the 36 wells around the plate are filled with 100 μL of PBS per well. Culture the cells in a 37°C, 5% CO2 incubator.

[0114] 3. Perform transient transfection experiments 24 hours after cell plating. Prepare transient transfection plasmids (i.e. Gal4-RORγ-LBD: 25ng / well; PgL4.3-luc: 25ng / well) and transfection reagents (the concentration of liposome 2000 is 3 times that of DNA).

[0115] 4. After the transfection reagent is diluted, incubate for 5 minutes, mix the transfection reagent and plasmid and incubate for 20 minutes, then add 10 μL to each well. If the transient transfection lasts for more than 5 hours, small molecule compounds (SR1001 or Exp1-Exp34 compounds) can be added.

[0116] 5. If necessary, dilute the compound to be tested 3-fold (100-0.195 μM) with DMEM cell culture medium containing 10% fetal bovine serum, then aspirate the existing culture medium in the cell culture plate, and then add the prepared compound to be tested and fresh culture medium.

[0117] 6. Then place the cells in a 37°C, 5% CO2 incubator for culture. After about 24 hours, take out the cells, observe the cell growth under a microscope, and take the cell culture plate out of the cell room. Then perform a luciferase dual reporter gene detection experiment.

[0118] 7. First, discard the cell culture medium by aspirating, and then add about 100 μL PBS to wash the residual culture medium. Dilute the 5× cell lysis buffer to 1×, add 20 μL to each well, and then shake for about 20 minutes to lyse the cells.

[0119] 8. Transfer the cells into a white opaque 96-well assay plate. Then use the En Spire Alpha 2390 homogeneous luminescent immunoassay system to test the experimental results: Add the prepared firefly luciferin substrate to detect the cell activity after the compound interference.

[0120] 9. Calculation of inhibitory activity:

[0121]

[0122] 10. Experimental results:

[0123] Compound <![CDATA[Half maximal inhibitory concentration (IC 50 )]]> Exp 1 ++++ Exp 2 ++++ Exp 3 +++ Exp 4 ++ Exp 5 ++++ Exp 6 +++ Exp 7 ++++ Exp 8 + SR1001 21510

[0124] ++++ indicates IC 50 <10nM; +++ indicates IC 50 The range is 10 to 100 nM; ++ indicates IC 50 The range is 100 to 1000 nM; + indicates IC 50 >1000nM.

[0125] Example 10 RORγt binding experiment

[0126] 1. Reagents and consumables:

[0127]

[0128] 2. Compound Management:

[0129] 2.1 Compound storage: Compounds were dissolved in DMSO to make 10 mM stock solutions.

[0130] 2.2 Compound storage: All compounds dissolved in DMSO should be stored in a desiccator at room temperature for no more than 3 months in the short term and at -20°C in the long term.

[0131] 2.3 Preparation of compounds:

[0132] a) All compounds were diluted 3-fold with DMSO, with a starting concentration of 500uM.

[0133] b) The positive control compound was diluted 3-fold with DMSO, with a 10-dilution gradient and a starting concentration of 25 uM.

[0134] c) Prepare 50x positive control (25uM positive control compound) and 50x negative control (100% DMSO).

[0135] d) Block the compound plate and shake for 5 minutes.

[0136] 3. Experimental process:

[0137] 3.1 Prepare reaction buffer: Dissolve DTT and KF in 1x buffer D. Final concentration: DTT 5mM, KF 50mM.

[0138] 3.2 Test compounds:

[0139] a) Prepare 2x serial dilutions of the compounds in buffer (see step 2.3).

[0140] b) Add 10ul of 2x serial dilutions of compound (see step a) to a 384-well reaction plate.

[0141] c) Prepare 2x reaction mixture in chilled buffer: RORγ-LBD (40 nM), SRC (100 nM), anti-GST Eu (1:200) and streptavidin-D2 (25 nM).

[0142] d) Add 10ul of 2x reactants (see step c) to the 384-well reaction plate (see step b).

[0143] e) Centrifuge the 384-well reaction plate at 1000 g for 1 min.

[0144] f) Incubate at room temperature in the dark for 1 hour.

[0145] g) Detection plate: wavelength 665nm and 615nm; instrument: multi-label microplate reader.

[0146] 4. Data Analysis

[0147] 4.1 Relative ratio (RR): Calculate the relative ratio of each well [(665nm response value / 615nm response value-blank background response value)*1000].

[0148] 4.2 The calculation of percent inhibition (% Inhibition) is as follows:

[0149]

[0150] 4.3 Calculation of IC of compounds 50 And dose-effect curve: The IC50 and dose-effect curve of the compound were obtained by calculating the inhibition rate of the compound and the log value of the compound concentration using Graphpad 5.0.

[0151] 4.4 Inspection Report:

[0152] 4.4.1 One experimenter completes the report and another experimenter checks the report again to ensure the accuracy of the data.

[0153] 4.4.1.1 Data are exported from the test instrument and analyzed manually.

[0154] 4.4.1.2 Convert the ratio to the percentage inhibition rate. The IC value of the compound was calculated for the first time using Graphpad5.0 software and the percentage inhibition rate. 50 .

[0155] 4.4.1.3 Calculating the IC of a Compound Using Ratios 50 , using this IC 50Check the data for accuracy.

[0156] 4.4.2 Determine whether the names of all compounds are correct.

[0157] 4.5 Data standard: Z factor>0.5; S / B>3;

[0158] IC of positive control compound 50 Within 3 times of the historical average.

[0159] 5. Data results:

[0160] Compound <![CDATA[Half maximal inhibitory concentration (IC 50 )]]> Exp 1 20nM Exp 2 14nM Exp 5 5nM Exp 6 50nM Exp 8 187nM SR1001 295nM

[0161] It can be seen from the above experimental results that the compound of formula (I) has a significant inhibitory effect on RORγt, and RORγt plays a very important role in inflammatory, metabolic and autoimmune diseases. Inhibition of RORγt will alleviate or effectively treat these diseases. In particular, the use of RORγt inhibitors in the treatment of respiratory diseases (such as asthma, COPD) and autoimmune diseases (such as rheumatoid arthritis, psoriasis, ulcerative colitis, Crohn's disease) has been intensively studied and recognized.

[0162] Although the present invention has been disclosed as above in terms of preferred embodiments, they are not intended to limit the present invention. Anyone skilled in the art can make various changes or modifications without departing from the spirit and scope of the present invention. Therefore, the scope of protection of the present invention should be based on the scope of protection defined by the claims of this application.

Claims

1. A compound having a structure as shown in formula (I) or a stereoisomer, a tautomer or a pharmaceutically acceptable salt or a solvate or a prodrug thereof: in, R1 is selected from hydrogen, halogen, C1-C3 alkyl, C1-C3 haloalkyl, alkoxy; X is selected from N or CR6, R6 is selected from halogen, C1-C3 alkyl, C1-C3 haloalkyl, alkoxy; Ring A is a 5-10 membered heteroaryl or a 6-10 membered aromatic group; R2 is selected from a C1-C6 alkyl group, a C3-C8 cycloalkyl group, a 3-8 membered heterocycloalkyl group, an aryl group, a heteroaryl group, and a cyano group; R 3、 R4 is independently selected from hydrogen, halogen, C1-C6 alkyl and C1-C6 haloalkyl, or R 3、 R4 merges into a ring; Ring B is a 5-10 membered heterocycloalkyl, a 6-10 membered aryl or a 5-10 membered heteroaryl, and R5 is selected from halogen, hydroxy, amino, C3-C6 cycloalkyl or C1-C6 alkyl.

2. The compound according to claim 1, or its stereoisomer, tautomer, or pharmaceutically acceptable salt, or solvate, or prodrug, characterized in that: The R1 is selected from halogen, methyl, ethyl, propyl, isopropyl, n-butyl, tert-butyl, -CF3.

3. The compound according to claim 2, or its stereoisomer, tautomer, or pharmaceutically acceptable salt, or solvate, or prodrug, characterized in that: The X is selected from N or CR6, and R6 is Cl.

4. The compound according to any one of claims 1 to 3, or its stereoisomer, tautomer, or pharmaceutically acceptable salt, or solvate, or prodrug, characterized in that: The ring A is one of a benzene ring, a pyrimidine ring, and a thiazole ring, and the R2 is selected from a C1-C6 alkyl group and a cyano group.

5. The compound according to claim 4, or its stereoisomer, tautomer, or pharmaceutically acceptable salt, or solvate, or prodrug, characterized in that: The R3 and R4 are independently selected from hydrogen, halogen, methyl, ethyl, propyl, isopropyl, n-butyl and tert-butyl.

6. The compound according to claim 5, or its stereoisomer, tautomer, or pharmaceutically acceptable salt, or solvate, or prodrug, characterized in that: The ring B comprises: "*" is the connection position with N; R5 is C1-C6 alkyl, including methyl, ethyl, propyl, isopropyl, n-butyl and tert-butyl.

7. The compound according to claim 6, or its stereoisomer, tautomer, or pharmaceutically acceptable salt, or solvate, or prodrug, characterized in that: The compounds shown in formula (I) include:

8. Use of the compound according to any one of claims 1 to 7 or its stereoisomer, tautomer or pharmaceutically acceptable salt or solvate or prodrug thereof in the preparation of a medicament for treating RORγ-mediated diseases.

9. The use according to claim 8, characterized in that The disease is an inflammatory, metabolic or autoimmune disease, including asthma, chronic obstructive pulmonary disease, bronchitis, allergic rhinitis, atopic dermatitis, cystic fibrosis, lung allograft rejection, multiple sclerosis, rheumatoid arthritis, juvenile rheumatoid arthritis, osteoarthritis, ankylosing spondylitis, systemic lupus erythematosus, psoriasis, Hashimoto's disease, pancreatitis, autoimmune diabetes, autoimmune eye disease, ulcerative colitis, Crohn's disease, inflammatory bowel disease, inflammatory bowel syndrome, Sjögren's syndrome, optic neuritis, type I diabetes, neuromyelitis optica, myasthenia gravis, uveitis, Guillain-Barré syndrome, psoriatic arthritis, Graves' disease or scleritis.

Images