TW201900617A - Spirocyclic methotrexate derivatives, preparation method thereof and application thereof in medicine - Google Patents

Abstract

The present invention relates to a spirocycle formamide derivative, a preparation method thereof and a medical use thereof. In particular, the present invention relates to a spirocycle formamide derivative as shown by general formula (AI), a preparation method thereof and a pharmaceutical composition containing the derivative, as well as a use thereof as a ROR modulator in the prevention and/or treatment of diseases such as inflammation, autoimmune diseases and cancer. The substituents in the general formula (AI) are the same as those defined in the description.

Description

   Spirocarboxamide derivatives, preparation method and application in medicine   

The invention belongs to the field of medicine and relates to spirocarboxamide derivatives, a preparation method thereof and its application in medicine. In particular, the present invention relates to a spirocarboxamide derivative represented by the general formula (AI), a preparation method thereof, a pharmaceutical composition containing the derivative, a ROR modulator, and a preventive and / or therapeutic agent thereof Use in medicine for inflammation, autoimmune disease, tumor or cancer.

Retinoic acid-related orphan nuclear receptor (ROR) is a member of the nuclear receptor family, which can regulate a variety of physiological and life processes. The ROR family contains three types of RORα, RORβ, and RORγ. Three different RORs can be expressed in different tissues and regulate different physiological processes. RORα is mainly distributed in liver, skeletal muscle, skin, lung, adipose tissue, kidney, thymus and brain. RORβ has a small range of action and mainly acts on the center Nervous system, RORγ can be expressed in many tissues, including liver, animal fat and skeletal muscle. The lack of RORγ in mammals shows a decrease in blood glucose.

There are two subtypes of RORγ: RORγ1 and RORγ2. RORγ1 is expressed in many tissues, such as thymus, muscle, kidney and liver, while RORγ2 is only expressed in immune cells, and RORγ2 is thought to control T cell helper T cell 17 (Th17) differentiation. Th17 is a class of helper T cells that can produce interleukin 17 (IL-17) and other cytokines. Th-17 has been found to be involved in human inflammatory diseases and immune disorders, such as multiple sclerosis, rheumatism Arthritis, psoriasis, clonal disease, and asthma are related. Now it has been reported in the literature that RORγ may be related to the occurrence and development of prostate cancer.

RORγt is a subtype that is specifically expressed on immune cells by RORγ. It is a major transcription factor for human and mouse Th17 cells. It can not only promote Th17 cell differentiation, but also regulate the expression and secretion of specific effector factor IL-17 in Th17 cells. RORγt is closely related to the occurrence and development of various immune diseases, infectious diseases and tumors.

RORγ, especially the RORγt type, has been identified as an important transcriptional regulator of Th17 cell differentiation. A 2006 study by Vanov et al. Found that RORγt is an important transcription factor for Th17 cell differentiation in mouse experiments. Their research showed that mice lacking RORγt had difficulty in inducing an EAE model. In the process of human Th17 cell differentiation, RORγt was quickly confirmed to have a similarly important role. The groundbreaking discovery caused people to attach great importance to RORγt.

At present, ROR as an inhibitor has received high attention in the medical field and has become a hot issue for research. Patent applications that have been published include WO2015171610, WO2015171558, WO2015131035, WO2013169864, WO2014179564, WO2015116904, and so on.

In the process of studying the ROR modulator, the inventors discovered that in the compound represented by the general formula (I) according to the present invention, the change of the ortho group of ring A will change its regulating effect. When the group is a group with a small steric hindrance (such as H), the compound represented by the general formula (I) is an inhibitor, and when the ortho group of the ring A is a haloalkyl group (such as a trifluoromethyl group), the steric hindrance is large. When the group represented by the general formula (I) is a ROR agonist, the present invention has developed a new generation of ROR modulators, and further studies have found that changes in the structure of the compounds can regulate different mechanisms.

The object of the present invention is to provide a compound represented by the general formula (AI): Or tautomers, mesomers, racemates, enantiomers, diastereomers or mixtures thereof, or a pharmaceutically acceptable salt thereof, wherein: W ¹ , W ² and W ^{3 is the} same or different, and each is independently CH or N; ring A and ring B are the same or different, and each is independently selected from cycloalkyl, heterocyclyl, aryl, and heteroaryl; ring C is cycloalkane Or heterocyclyl; R ^{1 is the} same or different, and each is independently selected from the group consisting of a hydrogen atom, halogen, alkyl, haloalkyl, alkoxy, haloalkoxy, cyano, amino, nitro, hydroxyl, Hydroxyalkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl, wherein the alkyl, haloalkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl are each independently selected from the group as needed One of halogen, alkyl, haloalkyl, alkoxy, haloalkoxy, cyano, amine, nitro, hydroxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl Or two or more substituents; R ^{2 is the} same or different, and each is independently selected from a hydrogen atom, halogen, alkyl, haloalkyl, alkoxy, haloalkoxy, cyano, amino, nitro, Hydroxy, hydroxyalkyl , Cycloalkyl, heterocyclyl, aryl, heteroaryl, -OR ⁴ , -C (O) R ⁴ , -C (O) OR ⁴ and -S (O) _m R ⁴ ; R ^{3 is} selected from hydrogen Atom, halogen, alkyl, haloalkyl, alkoxy, haloalkoxy, cyano, amine, nitro, hydroxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl, Wherein the alkyl group, haloalkyl group, cycloalkyl group, heterocyclic group, aryl group and heteroaryl group are each independently selected from hydroxy, halogen, alkyl, haloalkyl, cyano, amine, nitro, Substituted with one or more substituents of alkoxy, haloalkoxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl; R ^{4 is} selected from hydrogen atom, alkyl, cycloalkane Alkyl, haloalkyl, alkoxy, haloalkoxy, heterocyclyl, aryl, and heteroaryl, wherein the alkyl, cycloalkyl, haloalkyl, heterocyclyl, aryl, and heteroaryl are each Independently selected from the group consisting of hydroxy, halogen, alkyl, haloalkyl, cyano, amine, nitro, alkoxy, haloalkoxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl And heteroaryl substituted with one or more substituents; m is 0, 1 or 2; s is 0, 1, 2, 3, or 4; and t is 0, 1, 2, 3, or 4.

In a preferred embodiment of the present invention, the compound represented by the general formula (AI), wherein ring C is a C _3-6 cycloalkyl group or a 3- to 6-membered heterocyclic group, wherein the heterocyclic group contains 1 to 3 heteroatoms selected from N, O or S; preferably selected from cyclopropyl, cyclobutyl, cyclopentyl or tetrahydropyranyl.

In a preferred embodiment of the present invention, the compound represented by the general formula (AI) is a compound represented by the general formula (I): Or a tautomer, a racemate, a racemate, an enantiomer, a diastereomer or a mixture thereof, or a pharmaceutically acceptable salt thereof, wherein: n is 1, 2 or 3; Ring A, Ring B, R ¹ to R ³ , s and t are as defined in General Formula (AI).

In a preferred embodiment of the present invention, the compound represented by the general formula (I) is a compound represented by the general formula (II):

Wherein: ring A, ring B, R ¹ to R ³ , s and t are as defined in the general formula (I).

In a preferred embodiment of the present invention, the compound represented by the general formula (I), wherein ring A is selected from phenyl, pyridyl, pyrimidinyl, cyclohexyl, and piperidinyl.

In a preferred embodiment of the present invention, the compound represented by the general formula (I) is a compound represented by the general formula (III):

Wherein: G ¹ and G ^{2 are the} same or different, and are each independently CH or N; rings B, R ¹ to R ³ and t are as defined in the general formula (I).

In a preferred embodiment of the present invention, the compound represented by the general formula (I) is a compound represented by the general formula (IV):

Wherein: R ^{b is} selected from the group consisting of alkyl, haloalkyl, alkoxy, haloalkoxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl, wherein the alkyl, haloalkyl, Cycloalkyl, heterocyclyl, aryl, and heteroaryl are each independently selected from halogen, hydroxy, alkyl, haloalkyl, cyano, amine, nitro, alkoxy, haloalkoxy , Hydroxyalkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl with one or more substituents; p is 0, 1, 2 or 3; ring B, R ¹ ~ R ³ and t As defined in general formula (I).

In a preferred embodiment of the present invention, the compound represented by the general formula (I), wherein the ring B is selected from phenyl, pyridyl, pyrimidinyl, cyclohexyl, and piperidinyl.

In a preferred embodiment of the present invention, the compound represented by the general formula (I), wherein R ² is -S (O) _m R ⁴ ; m is 2; and R ⁴ is alkyl, preferably ethyl base.

In a preferred embodiment of the present invention, the compound represented by the general formula (III) is a compound represented by the general formula (III-A):

Wherein: R ¹ and R ³ to R ⁴ are as defined in the general formula (III).

In a preferred embodiment of the present invention, the compound represented by the general formula (IV) is a compound represented by the general formula (IV-A):

Where: R ^{b is} selected from the group consisting of alkyl, haloalkyl, alkoxy, haloalkoxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl, wherein the alkyl, haloalkyl, Cycloalkyl, heterocyclyl, aryl, and heteroaryl are each independently selected from halogen, hydroxy, alkyl, haloalkyl, cyano, amine, nitro, alkoxy, haloalkoxy , Hydroxyalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl with one or more substituents; p is 0, ¹ , 2 or 3; R ¹ , R ³ and R ^{4 are} as follows As defined in formula (IV).

In a preferred embodiment of the present invention, the compound represented by the general formula (III-A) or (IV-A), wherein R ⁴ is an alkyl group, preferably an ethyl group.

In a preferred embodiment of the present invention, the compound represented by the general formula (IV) or (IV-A), wherein R ^b is a haloalkyl group, and preferably trifluoromethyl.

In a preferred embodiment of the present invention, the compound represented by the general formula (I), wherein R ¹ is a haloalkyl group, and preferably a trifluoromethyl group.

In a preferred embodiment of the present invention, the compound represented by the general formula (I), wherein R ³ is a hydrogen atom, an alkyl group or a hydroxyalkyl group.

Typical compounds of general formula (I), including but not limited to:

Or a tautomer, a racemate, a racemate, an enantiomer, a diastereomer or a mixture thereof, or a pharmaceutically acceptable salt thereof.

In another aspect, the present invention provides a compound represented by the general formula (AI-A) for preparing a compound according to the general formula (AI) or a tautomer, meso, racemate, enantiomer thereof. Intermediates, isomers, diastereomers, or mixtures thereof, or pharmaceutically acceptable salts thereof:

Or a tautomer, a racemate, a racemate, an enantiomer, a diastereomer or a mixture thereof, or a pharmaceutically acceptable salt thereof, wherein: ring A, ring C, W ¹ , W ² , W ³ , R ¹ and s are as defined in the general formula (AI).

In another preferred embodiment, the present invention provides a compound represented by the general formula (IA) for preparing a compound according to the general formula (I) or a tautomer, a racemate, a racemate, Enantiomers, diastereomers, or a mixture thereof, or an intermediate of a pharmaceutically acceptable salt thereof:

Or a tautomer, a racemate, a racemate, an enantiomer, a diastereomer or a mixture thereof, or a pharmaceutically acceptable salt thereof, wherein: ring A, R ¹ , s And n are as defined in general formula (I).

Typical compounds of general formula (AI-A), including but not limited to:

In another aspect, the present invention provides a method for preparing the compound of the general formula (AI), the method comprising:

A compound of the general formula (AI-A) and a compound of the general formula (IB) undergo a condensation reaction to obtain a compound of the general formula (AI); wherein: ring A, ring B, ring C, W ¹ , W ² , W ³ , R ¹ ~ R ³ , s and t are as defined in the general formula (AI).

In another aspect, the present invention provides a method for preparing the compound of the general formula (I), the method comprising:

A compound of the general formula (IA) and a compound of the general formula (IB) undergo a condensation reaction to obtain a compound of the general formula (I); wherein: ring A, ring B, R ¹ to R ³ , n, s, and t are as shown in the general formula (I) As defined in.

In another aspect, the present invention provides a method for preparing the compound of general formula (II), the method comprising:

A compound of the general formula (II-A) and a compound of the general formula (IB) undergo a condensation reaction to obtain a compound of the general formula (II); wherein: ring A, ring B, R ¹ to R ³ , s, and t are as in the general formula (II) As defined in.

Another aspect of the present invention relates to a pharmaceutical composition containing a therapeutically effective dose of a compound represented by the general formula (I) or a tautomer, meso, racemate, enantiomer, Diastereomers, or a mixture thereof, or a pharmaceutically acceptable salt, and one or more pharmaceutically acceptable carriers, diluents, or excipients. The present invention also relates to a method for preparing the above composition, which comprises converting a compound represented by the general formula (I) or a tautomer, a racemate, a racemate, an enantiomer, a diastereomer The isomer, or a mixture thereof, or a pharmaceutically acceptable salt thereof is mixed with a pharmaceutically acceptable carrier, diluent, or excipient.

The present invention further relates to a compound represented by the general formula (I) or a tautomer, a racemate, a racemate, an enantiomer, a diastereomer or a mixture thereof, or a compound thereof Use of a pharmaceutically acceptable salt, or a pharmaceutical composition containing the same, in the preparation of a ROR modulator.

The present invention further relates to a compound represented by the general formula (I) or a tautomer, a racemate, a racemate, an enantiomer, a diastereomer or a mixture thereof, or a compound thereof Use of a pharmaceutically acceptable salt, or a pharmaceutical composition comprising the same, in the manufacture of a medicament for the prevention and / or treatment of inflammation, autoimmune disease, tumor or cancer.

The present invention further relates to a compound represented by the general formula (I) or a tautomer, a racemate, a racemate, an enantiomer, a diastereomer or a mixture thereof, or a compound thereof Use of a pharmaceutically acceptable salt, or a medicinal composition comprising the same, in the manufacture of a medicament for the prevention and / or treatment of inflammation, autoimmune disease, tumor or cancer, the inflammation or autoimmune disease selected from asthma, atopic Dermatitis, contact dermatitis, acne, bronchitis, Crohn's disease, localized enteritis, inflammatory bowel disease (IBD), ulcerative colitis, allograft rejection, Suglian's syndrome, uveitis , Behcet's disease, dermatomyositis, multiple sclerosis, ankylosing spondylitis, neuromyelitis, systemic lupus erythematosus (SLE), scleroderma, pancreatitis, psoriasis, psoriasis arthritis (PsA), rheumatoid arthritis, arthritis, osteoarthritis, allergic rhinitis, autoimmune diabetes and autoimmune thyroid disease, the tumor or cancer is selected from non-Hodgkin's lymphoma, diffuse large B-cell lymphoma Tumor, follicular lymphoma, slippery Sarcoma, breast cancer, cervical cancer, colon cancer, lung cancer, stomach cancer, rectal cancer, pancreatic cancer, brain cancer, skin cancer, oral cancer, prostate cancer, bone cancer, kidney cancer, ovarian cancer, bladder cancer, liver cancer, fallopian tube tumor, Ovarian tumor, peritoneal tumor, stage IV melanoma, solid tumor, glioma, glioblastoma, hepatocellular carcinoma, mastoid nephroma, head and neck tumor, leukemia, lymphoma, myeloma, and non-small cell lung cancer .

The present invention further relates to a compound represented by the general formula (I) or a tautomer, a racemate, a racemate, an enantiomer, a diastereomer or a mixture thereof, or a compound thereof Use of a pharmaceutically acceptable salt or a medicinal composition containing the same as a ROR inhibitor in the manufacture of a medicament for the prevention and / or treatment of inflammation and autoimmune diseases, the inflammation or autoimmune diseases selected from asthma, atopic Dermatitis, contact dermatitis, acne, bronchitis, Crohn's disease, localized enteritis, inflammatory bowel disease (IBD), ulcerative colitis, allograft rejection, Suglian's syndrome, uveitis , Behcet's disease, dermatomyositis, multiple sclerosis, ankylosing spondylitis, neuromyelitis, systemic lupus erythematosus (SLE), scleroderma, pancreatitis, psoriasis, psoriasis arthritis (PsA), rheumatoid arthritis, arthritis, osteoarthritis, allergic rhinitis, autoimmune diabetes and autoimmune thyroid disease.

The present invention further relates to a compound represented by the general formula (I) or a tautomer, a racemate, a racemate, an enantiomer, a diastereomer or a mixture thereof, or a compound thereof Use of a pharmaceutically acceptable salt, or a pharmaceutical composition containing the same, as a ROR agonist in the manufacture of a medicament for the prevention and / or treatment of a tumor or cancer selected from non-Hodgkin's lymphoma, diffuse large B Cellular lymphoma, follicular lymphoma, synovial sarcoma, breast cancer, cervical cancer, colon cancer, lung cancer, gastric cancer, rectal cancer, pancreatic cancer, brain cancer, skin cancer, oral cancer, prostate cancer, bone cancer, kidney cancer , Ovarian cancer, bladder cancer, liver cancer, fallopian tube tumor, ovarian tumor, peritoneal tumor, stage IV melanoma, solid tumor, glioma, glioblastoma, hepatocellular carcinoma, mastoid nephroma, head and neck tumor, Leukemia, lymphoma, myeloma, and non-small cell lung cancer.

The present invention further relates to a compound represented by the general formula (I) or a tautomer, a racemate, a racemate, an enantiomer, a diastereomer or a mixture thereof, or A pharmaceutically acceptable salt, which is used as a medicament.

The present invention also relates to a compound represented by the general formula (I) or a tautomer, a racemate, a racemate, an enantiomer, a diastereomer or a mixture thereof, or A pharmaceutically acceptable salt, or a pharmaceutical composition containing the same, which acts as a ROR modulator.

The present invention also relates to a compound represented by the general formula (I) or a tautomer, a racemate, a racemate, an enantiomer, a diastereomer or a mixture thereof, or A pharmaceutically acceptable salt, or a pharmaceutical composition containing the same, for the prevention and / or treatment of inflammation, autoimmune disease, tumor or cancer.

The present invention also relates to a compound represented by the general formula (I) or a tautomer, a racemate, a racemate, an enantiomer, a diastereomer or a mixture thereof, or A pharmaceutically acceptable salt, or a pharmaceutical composition containing the same, for preventing and / or treating an inflammation or an autoimmune disease selected from asthma, atopic dermatitis, contact dermatitis, acne, Bronchitis, Crohn's disease, local enteritis, inflammatory bowel disease (IBD), ulcerative colitis, allograft rejection, Suglian's syndrome, uveitis, Behcet's disease, skin muscle Inflammation, multiple sclerosis, ankylosing spondylitis, neuromyelitis, systemic lupus erythematosus (SLE), scleroderma, pancreatitis, psoriasis, psoriatic arthritis (PsA), rheumatoid joints Inflammation, arthritis, osteoarthritis, allergic rhinitis, autoimmune diabetes and autoimmune thyroid disease.

The present invention also relates to a compound represented by the general formula (I) or a tautomer, a racemate, a racemate, an enantiomer, a diastereomer or a mixture thereof, or A pharmaceutically acceptable salt, or a pharmaceutical composition comprising the same, for preventing and / or treating a tumor or cancer selected from non-Hodgkin's lymphoma, diffuse large B-cell lymphoma, follicular lymphoma, synovial Mesosarcoma, breast cancer, cervical cancer, colon cancer, lung cancer, stomach cancer, rectal cancer, pancreatic cancer, brain cancer, skin cancer, oral cancer, prostate cancer, bone cancer, kidney cancer, ovarian cancer, bladder cancer, liver cancer, fallopian tube tumor , Ovarian tumor, peritoneal tumor, stage IV melanoma, solid tumor, glioma, glioblastoma, hepatocellular carcinoma, mastoid nephroma, head and neck tumor, leukemia, lymphoma, myeloma, and non-small cells Lung cancer.

The present invention also relates to a method for preventing and / or treating inflammation, autoimmune disease, tumor or cancer, which comprises administering to a patient in need thereof a therapeutically effective dose of a compound represented by the general formula (I) or a tautomer thereof , Meso, racemate, enantiomer, diastereomer or mixture thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing the same, the inflammation or autoimmune disease Selected from asthma, atopic dermatitis, contact dermatitis, acne, bronchitis, Crohn's disease, local enteritis, inflammatory bowel disease (IBD), ulcerative colitis, allograft rejection, Hugh Glen Syndrome, uveitis, Behcet's disease, dermatomyositis, multiple sclerosis, ankylosing spondylitis, neuromyelitis, systemic lupus erythematosus (SLE), scleroderma, pancreatitis, silver Psoriasis, psoriatic arthritis (PsA), rheumatoid arthritis, arthritis, osteoarthritis, allergic rhinitis, autoimmune diabetes and autoimmune thyroid disease, said tumor or cancer is selected from non-Hodge Gold lymphoma, diffuse large B fine Lymphoma, follicular lymphoma, synovial sarcoma, breast cancer, cervical cancer, colon cancer, lung cancer, gastric cancer, rectal cancer, pancreatic cancer, brain cancer, skin cancer, oral cancer, prostate cancer, bone cancer, kidney cancer, Ovarian cancer, bladder cancer, liver cancer, fallopian tube tumor, ovarian tumor, peritoneal tumor, stage IV melanoma, solid tumor, glioma, glioblastoma, hepatocellular carcinoma, mastoid nephroma, head and neck tumor, leukemia , Lymphoma, myeloma, and non-small cell lung cancer.

The present invention also relates to a method for preventing and / or treating inflammation or an autoimmune disease, which comprises administering to a patient in need thereof a therapeutically effective dose of a compound represented by the general formula (I) as a ROR inhibitor or a tautomer thereof , Meso, racemate, enantiomer, diastereomer or mixture thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing the same, the inflammation or autoimmunity The disease is selected from asthma, atopic dermatitis, contact dermatitis, acne, bronchitis, Crohn's disease, local enteritis, inflammatory bowel disease (IBD), ulcerative colitis, allograft rejection, Hugh Len's syndrome, uveitis, Behcet's disease, dermatomyositis, multiple sclerosis, ankylosing spondylitis, neuromyelitis, systemic lupus erythematosus (SLE), scleroderma, pancreatitis, Psoriasis, psoriatic arthritis (PsA), rheumatoid arthritis, arthritis, osteoarthritis, allergic rhinitis, autoimmune diabetes, and autoimmune thyroid disease.

The present invention also relates to a method for treating prevention and / or treatment of tumor or cancer, which comprises administering to a patient in need thereof a therapeutically effective dose of a compound represented by general formula (I) or a tautomer thereof as a ROR agonist, Meso, racemate, enantiomer, diastereomer or mixture thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the same, the tumor or cancer is selected from non-Ho Chitin lymphoma, diffuse large B-cell lymphoma, follicular lymphoma, synovial sarcoma, breast cancer, cervical cancer, colon cancer, lung cancer, gastric cancer, rectal cancer, pancreatic cancer, brain cancer, skin cancer, oral cancer, Prostate cancer, bone cancer, kidney cancer, ovarian cancer, bladder cancer, liver cancer, fallopian tube tumor, ovarian tumor, peritoneal tumor, stage IV melanoma, solid tumor, glioma, glioblastoma, hepatocellular carcinoma, mastoid Renal tumors, head and neck tumors, leukemia, lymphoma, myeloma, and non-small cell lung cancer.

The present invention also relates to a method for regulating ROR, which comprises administering to a patient in need thereof a therapeutically effective dose of a compound represented by general formula (I) or a tautomer, meso, racemate, enantiomer thereof Isomers, diastereomers, or a mixture thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing the same.

The active ingredient-containing pharmaceutical composition may be in a form suitable for oral administration, such as tablets, dragees, lozenges, water or oil suspensions, dispersible powders or granules, emulsions, hard or soft capsules, or syrups or Tincture. Oral compositions may be prepared according to any method known in the art for preparing pharmaceutical compositions, and such compositions may contain one or more ingredients selected from the group consisting of sweeteners, flavoring agents, colorants, and preservatives to provide pleasing looks And delicious medicinal preparations. Tablets contain the active ingredients and non-toxic pharmaceutically acceptable excipients suitable for the preparation of tablets for mixing. These excipients can be inert excipients, granulating agents, disintegrating agents, binders, and lubricants. These tablets can be uncoated or they can be coated by known techniques that mask the taste of the drug or delay disintegration and absorption in the gastrointestinal tract, thereby providing a sustained release over a longer period.

Oral formulations may also be provided in soft gelatin capsules in which the active ingredient is mixed with an inert solid diluent or in which the active ingredient is mixed with a water-soluble carrier or an oil vehicle.

Aqueous suspensions contain the active substance and excipients suitable for the preparation of the aqueous suspension for mixing. Such excipients are suspending, dispersing or wetting agents. The aqueous suspension may also contain one or more preservatives, one or more coloring agents, one or more flavoring agents, and one or more sweetening agents.

Oil suspensions can be formulated by suspending the active ingredient in a vegetable or mineral oil. The oil suspension may contain a thickener. The sweeteners and flavoring agents described above can be added to provide a palatable formulation. These compositions can be preserved by the addition of antioxidants.

The pharmaceutical composition of the present invention may also be in the form of an oil-in-water emulsion. The oil phase may be a vegetable oil, or a mineral oil or a mixture thereof. Suitable emulsifiers may be naturally occurring phospholipids, and emulsions may also contain sweeteners, flavoring agents, preservatives and antioxidants. Such formulations may also contain a demulcent, a preservative, a colorant, and an antioxidant.

The pharmaceutical composition of the present invention may be in the form of a sterile injectable aqueous solution. Among the acceptable vehicles or solvents that may be employed are water, Ringer's solution and isotonic sodium chloride solution. The sterile injectable preparation may be a sterile injectable oil-in-water microemulsion in which the active ingredient is dissolved in the oil phase, and the injection solution or microemulsion may be injected into the bloodstream of the patient by local large-volume injection. Alternatively, solutions and microemulsions are preferably administered in a manner that maintains a constant circulating concentration of a compound of the invention. To maintain this constant concentration, continuous intravenous drug delivery devices can be used. An example of such a device is the Deltec CADD-PLUS.TM. 5400 intravenous pump.

The pharmaceutical composition of the present invention may be in the form of a sterile injectable water or oil suspension for intramuscular and subcutaneous administration. This suspension may be formulated according to the known art using those suitable dispersing or wetting agents and suspending agents which have been mentioned above. The sterile injectable preparation may also be a sterile injectable solution or suspension prepared in a parenterally acceptable non-toxic diluent or solvent. In addition, a sterile fixed oil can be conveniently used as a solvent or suspension medium. For this purpose, any blending fixing oil can be used. In addition, fatty acids can also be prepared for injection.

The compounds of the invention may be administered in the form of suppositories for rectal administration. These pharmaceutical compositions can be prepared by mixing the drug with a suitable non-irritating excipient that is solid at ordinary temperatures but liquid in the rectum and therefore will dissolve in the rectum to release the drug.

As known to those skilled in the art, the dosage of a drug depends on a number of factors, including but not limited to the following: the activity of the specific compound used, the age of the patient, the weight of the patient, the patient's health, the patient's behavior, Patient's diet, time of administration, mode of administration, rate of excretion, combination of drugs, etc .; In addition, the best treatment such as the mode of treatment, the daily dosage of the general compound (I) or the type of pharmaceutically acceptable salt Can be verified according to the traditional treatment plan.

    Detailed description of the invention    

Unless stated to the contrary, terms used in the specification and claims have the following meanings.

The term "alkyl" refers to a saturated aliphatic hydrocarbon group, which is a straight or branched chain group containing 1 to 20 carbon atoms, preferably an alkyl group containing 1 to 12 carbon atoms, more preferably 1 to 6 Carbon atom alkyl. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, third butyl, second butyl, n-pentyl, 1,1-dimethylpropyl , 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl- 2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1 , 3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, n-heptyl , 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2,3-dimethylpentyl, 2,4-dimethylpentyl, 2,2-di Methylpentyl, 3,3-dimethylpentyl, 2-ethylpentyl, 3-ethylpentyl, n-octyl, 2,3-dimethylhexyl, 2,4-dimethylhexyl , 2,5-dimethylhexyl, 2,2-dimethylhexyl, 3,3-dimethylhexyl, 4,4-dimethylhexyl, 2-ethylhexyl, 3-ethylhexyl, 4 -Ethylhexyl, 2-methyl-2-ethylpentyl, 2-methyl-3-ethylpentyl, n-nonyl, 2-methyl-2-ethylhexyl, 2-methyl-3 -Ethylhexyl , 2,2-diethylpentyl, n-decyl, 3,3-diethylhexyl, 2,2-diethylhexyl, and various branched isomers thereof. More preferred are lower alkyl groups containing 1 to 6 carbon atoms. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, third butyl, Dibutyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl Methyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2- Dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methyl Pentyl, 2,3-dimethylbutyl and the like. The alkyl group may be substituted or unsubstituted. When substituted, the substituent may be substituted at any available point of attachment. The substituent is preferably one or more of the following groups, which are independently selected from alkane Alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, ring Alkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, pendant oxygen, carboxyl, carboxylate, -OR ⁴ , -C (O) R ⁴ , -C (O) OR ⁴ and -S (O) _m R ⁴ .

The term "alkoxy" refers to -O- (alkyl) and -O- (unsubstituted cycloalkyl), wherein alkyl and cycloalkyl are as defined above. Non-limiting examples of alkoxy include: methoxy, ethoxy, propoxy, butoxy, cyclopropoxy, cyclobutoxy, cyclopentyloxy, cyclohexyloxy. The alkoxy group may be optionally substituted or unsubstituted. When substituted, the substituent is preferably one or more of the following groups, which are independently selected from the group consisting of alkyl, alkenyl, alkynyl, alkoxy, Alkylthio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkane Thio, heterocycloalkylthio, carboxyl, carboxylate, -OR ⁴ , -C (O) R ⁴ , -C (O) OR ⁴ and -S (O) _m R ⁴ .

The term "cycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent. The cycloalkyl ring contains 3 to 20 carbon atoms, preferably contains 3 to 12 carbon atoms, and more preferably contains 3 to 6 carbon atoms. Non-limiting examples of monocyclic cycloalkyl include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatriene Groups, cyclooctyl and the like; polycyclic cycloalkyl groups include spiro, fused and bridged cycloalkyl.

The term "spirocycloalkyl" refers to a polycyclic group with 5 to 20 members of a single ring sharing a carbon atom (called a spiro atom), which may contain one or more double bonds, but none of the rings are fully conjugated Π electronic system. It is preferably 6 to 14 members, more preferably 7 to 10 members. Spirocycloalkyl is divided into monospirocycloalkyl, bispirocycloalkyl or polyspirocycloalkyl according to the number of common spiro atoms between the rings, preferably monospirocycloalkyl and bispirocycloalkyl . More preferably, it is 4/4 members, 4 members / 5 members, 4 members / 6 members, 5 members / 5 members, or 5 members / 6 members monospirocycloalkyl. Non-limiting examples of spirocycloalkyl include:

The term "fused cycloalkyl" refers to a 5 to 20 member, each ring in the system and the other rings in the system share a pair of full carbon polycyclic groups adjacent to each other, where one or more rings may contain one or Multiple double bonds, but none of the rings have a completely conjugated π-electron system. It is preferably 6 to 14 members, more preferably 7 to 10 members. It can be divided into bicyclic, tricyclic, tetracyclic or polycyclic fused cycloalkyl according to the number of constituent rings, preferably bicyclic or tricyclic, more preferably 5-membered / 5-membered or 5-membered / 6-membered bicyclic alkyl. Non-limiting examples of fused cycloalkyl include:

The term "bridged cycloalkyl" refers to a full-carbon polycyclic group of 5 to 20 members in which any two rings share two carbon atoms that are not directly connected, which may contain one or more double bonds, but no ring has a complete Conjugate π electron system. It is preferably 6 to 14 members, more preferably 7 to 10 members. It can be divided into bicyclic, tricyclic, tetracyclic or polycyclic bridged cycloalkyl according to the number of constituent rings, preferably bicyclic, tricyclic or tetracyclic, and more preferably bicyclic or tricyclic. Non-limiting examples of bridged cycloalkyl include:

The cycloalkyl ring may be fused to an aryl, heteroaryl or heterocycloalkyl ring, wherein the ring connected to the parent structure is a cycloalkyl group, and non-limiting examples include indanyl, tetrahydronaphthyl , Benzocycloheptyl and the like. A cycloalkyl group may be optionally substituted or unsubstituted. When substituted, the substituent is preferably one or more of the following groups, which are independently selected from alkyl, alkenyl, alkynyl, alkoxy, Alkylthio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkane Thio, heterocycloalkylthio, pendant oxy, carboxyl, carboxylate, -OR ⁴ , -C (O) R ⁴ , -C (O) OR ⁴ and -S (O) _m R ⁴ .

The term "heterocyclyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent that contains 3 to 20 ring atoms, one or more of which are selected from nitrogen, oxygen, or S (O) A heteroatom of _m (where m is an integer from 0 to 2), excluding the ring portion of -OO-, -OS-, or -SS-, and the remaining ring atoms are carbon. Preferably contains 3 to 12 ring atoms, of which 1 to 4 are heteroatoms; most preferably contains 3 to 8 ring atoms, of which 1 to 3 are heteroatoms; most preferably contains 3 to 6 ring atoms, of which 1 to 2 is a hetero atom. Non-limiting examples of monocyclic heterocyclyl include pyrrolidinyl, imidazolidinyl, tetrahydrofuryl, tetrahydrothienyl, dihydroimidazolyl, dihydrofuryl, dihydropyrazolyl, dihydropyrrolyl, piperidine Group, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, pyranyl, and the like, piperidinyl, piperazinyl, or morpholinyl is preferred. Polycyclic heterocyclic groups include spiro, fused and bridged heterocyclic groups.

The term "spiroheterocyclyl" refers to a polycyclic heterocyclic group in which 5 to 20 members of a single ring share one atom (called a spiro atom), wherein one or more ring atoms are selected from nitrogen, oxygen, or S (O ) _m (where m is an integer from 0 to 2) and the remaining ring atoms are carbon. It can contain one or more double bonds, but none of the rings have a completely conjugated π-electron system. It is preferably 6 to 14 members, more preferably 7 to 10 members. Spiro heterocyclyl is divided into monospiroheterocyclyl, bispiroheterocyclyl or polyspiroheterocyclyl according to the number of common spiro atoms between the rings, preferably monospiroheterocyclyl and bispiroheterocyclyl . More preferred are 3 member / 6 member, 4 member / 4 member, 4 member / 5 member, 4 member / 6 member, 5 member / 5 member or 5 member / 6 member monospirocyclic group. Non-limiting examples of spiroheterocyclyl include:

The term "fused heterocyclyl" refers to a polycyclic heterocyclic group of 5 to 20 members. Each ring in the system shares an adjacent pair of atoms with other rings in the system. One or more rings may contain one or more Double bonds, but none of the rings have a completely conjugated π-electron system in which one or more ring atoms are heteroatoms selected from nitrogen, oxygen, or S (O) _m (where m is an integer from 0 to 2), and the remaining rings Atoms are carbon. It is preferably 6 to 14 members, more preferably 7 to 10 members. According to the number of constituent rings, it can be divided into bicyclic, tricyclic, tetracyclic or polycyclic fused heterocyclic group, preferably bicyclic or tricyclic, more preferably 5 member / 5 member or 5 member / 6 member bicyclic fused heterocyclic group . Non-limiting examples of fused heterocyclyl include:

The term "bridged heterocyclyl" refers to a polycyclic heterocyclic group of 5 to 14 members in which any two rings share two atoms that are not directly connected, which may contain one or more double bonds, but no ring has a complete A y-electron system of a yoke in which one or more ring atoms are heteroatoms selected from nitrogen, oxygen, or S (O) _m (where m is an integer of 0 to 2), and the remaining ring atoms are carbon. It is preferably 6 to 14 members, more preferably 7 to 10 members. It can be divided into bicyclic, tricyclic, tetracyclic or polycyclic bridge heterocyclic groups according to the number of constituent rings, preferably bicyclic, tricyclic or tetracyclic, more preferably bicyclic or tricyclic. Non-limiting examples of bridged heterocyclyls include:

The heterocyclyl ring may be fused to an aryl, heteroaryl or cycloalkyl ring, wherein the ring connected to the parent structure is a heterocyclic group, and non-limiting examples thereof include: with Wait.

The heterocyclic group may be optionally substituted or unsubstituted. When substituted, the substituent is preferably one or more of the following groups, which are independently selected from alkyl, alkenyl, alkynyl, alkoxy, Alkylthio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkane Thio, heterocycloalkylthio, pendant oxy, carboxyl, carboxylate, -OR ⁴ , -C (O) R ⁴ , -C (O) OR ⁴ and -S (O) _m R ⁴ .

The term "aryl" refers to a 6 to 14 membered all-carbon monocyclic or fused polycyclic (i.e., ring sharing a pair of adjacent carbon atom pairs) group, preferably 6 to 10 members, having a conjugated pi-electron system, such as Phenyl and naphthyl. Better phenyl. The aryl ring may be fused to a heteroaryl, heterocyclic or cycloalkyl ring, wherein the ring connected to the parent structure is an aryl ring, and non-limiting examples include:

The aryl group may be substituted or unsubstituted. When substituted, the substituent is preferably one or more of the following groups, which are independently selected from the group consisting of alkyl, alkenyl, alkynyl, alkoxy, and alkylthio , Alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, Heterocycloalkylthio, carboxyl or carboxylate.

The term "heteroaryl" refers to a heteroaromatic system containing 1 to 4 heteroatoms, 5 to 14 ring atoms, wherein the heteroatoms are selected from oxygen, sulfur, and nitrogen. Heteroaryl is preferably 5 to 10 members, containing 1 to 3 heteroatoms; more preferably 5 or 6 members, containing 1 to 2 heteroatoms; preferably, for example, imidazolyl, furanyl, thienyl, thiazole , Pyrazolyl, oxazolyl, pyrrolyl, tetrazolyl, pyridyl, pyrimidinyl, thiadiazole, pyrazinyl, etc., preferably imidazolyl, tetrazolyl, pyridyl, thienyl, pyrazole Or pyrimidinyl, thiazolyl; more preferably pyridyl. The heteroaryl ring may be fused to an aryl, heterocyclic or cycloalkyl ring, wherein the ring connected to the parent structure is a heteroaryl ring, and non-limiting examples include:

Heteroaryl may be optionally substituted or unsubstituted. When substituted, the substituent is preferably one or more of the following groups, which are independently selected from alkyl, alkenyl, alkynyl, alkoxy, Alkylthio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkane Thio, heterocycloalkylthio, carboxyl, carboxylate, -OR ⁴ , -C (O) R ⁴ , -C (O) OR ⁴ and -S (O) _m R ⁴ .

The term "haloalkyl" refers to an alkyl group substituted with one or more halogens, wherein alkyl is as defined above.

The term "haloalkoxy" refers to an alkoxy group substituted with one or more halogens, wherein alkoxy is as defined above.

The term "hydroxyalkyl" refers to an alkyl group substituted with a hydroxy group, wherein the alkyl group is as defined above.

The term "hydroxy" refers to the -OH group.

The term "halogen" refers to fluorine, chlorine, bromine or iodine.

The term "amino" refers to -NH _2.

The term "cyano" refers to -CN.

The term "nitro" refers to -NO _2.

The term "lateral oxygen" refers to = O.

The term "carbonyl" refers to C = O.

The term "carboxy" refers to -C (O) OH.

The term "carboxylate" refers to -C (O) O (alkyl) or -C (O) O (cycloalkyl), wherein alkyl and cycloalkyl are as defined above.

The term "halogen halide" refers to a compound containing a -C (O) -halogen group.

"As needed" or "as needed" means that the event or environment described later can, but does not have to occur, and the description includes situations where the event or environment occurs or does not occur. For example, "heterocyclic group substituted with an alkyl group as necessary" means that the alkyl group may but need not exist, and the description includes a case where the heterocyclic group is substituted with an alkyl group and a case where the heterocyclic group is not substituted with an alkyl group .

"Substituted" refers to one or more hydrogen atoms in a group, preferably up to 5 and more preferably 1 to 3 hydrogen atoms independently of one another by a corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and those skilled in the art can determine (by experiment or theory) possible or impossible substitutions without undue effort. For example, an amine or hydroxyl group having free hydrogen may be unstable when combined with a carbon atom having an unsaturated (eg, olefinic) bond.

"Pharmaceutical composition" means a mixture containing one or more of the compounds described herein, or a physiological / pharmaceutically acceptable salt or prodrug thereof, with other chemical components, and other components such as physiological / pharmaceutically acceptable carriers And excipients. The purpose of the pharmaceutical composition is to promote the administration to the living body, which is beneficial to the absorption of the active ingredient and then exerts the biological activity.

"Pharmaceutically acceptable salt" refers to a salt of a compound of the present invention. Such salts are safe and effective when used in mammals, and have due biological activity.

R ⁴ and m are as defined in the general formula (I).

    Method for synthesizing compounds of the present invention    

In order to achieve the purpose of the present invention, the present invention adopts the following technical scheme: the compound represented by the general formula (AI) of the present invention or a tautomer, meso, racemate, enantiomer, diastereomer A method for preparing an enantiomer or a mixture thereof, or a pharmaceutically acceptable salt thereof, includes the following steps:

In the first step, the compound of the general formula (AI-1) and paraformaldehyde are ring closed under acidic conditions to obtain the general formula (AI-2); in the second step, the compound of the general formula (AI-2) In the presence of a catalyst, it is reacted with CO and R ^c OH to obtain the general formula (AI-3). In the third step, the compound of the general formula (AI-3) is stripped of trifluoroacetamidine under basic conditions to obtain the general formula (AI- 4); In the fourth step, a compound of the general formula (AI-4) is reacted with a compound of the general formula (I-5) to obtain a compound of the general formula (AI-5) under basic conditions in the presence of a catalyst; or A compound of the general formula (I-4) is subjected to reductive amination with a compound of the general formula (I-6) under acidic conditions in the presence of a reducing agent to obtain a compound of the general formula (AI-5); in the fifth step, the general formula (AI -5) The compound is hydrolyzed to obtain (AI-A) compound under basic conditions; in the sixth step, the compound of general formula (AI-A) and compound of general formula (IB) occur under basic conditions in the presence of a condensing agent Condensation reaction to obtain compounds of general formula (AI); reagents providing basic conditions include organic bases and inorganic bases, the organic bases include but are not limited to triethylamine, N, N-diisopropylethylamine, n-butyl Lithium, lithium diisopropylamino, vinegar Potassium, sodium third butoxide or potassium third butoxide, the inorganic bases include but are not limited to sodium hydride, sodium hydroxide, potassium phosphate, sodium carbonate, sodium bicarbonate, potassium carbonate or cesium carbonate; Reagents include, but are not limited to, hydrogen chloride, 1,4-dioxane solution of hydrogen chloride, trifluoroacetic acid, formic acid, acetic acid, hydrochloric acid, sulfuric acid, methanesulfonic acid, nitric acid, phosphoric acid, p-benzenesulfonic acid, Me ₃ SiCl, and TMSOT _f ; Catalysts include but are not limited to palladium / carbon, Raney nickel, tetra-triphenylphosphine palladium, palladium dichloride, palladium acetate, 2-dicyclohexylphosphine-2,4,6-triisopropylbiphenyl, [1,1'-bis (diphenylphosphino) ferrocene] palladium dichloride, 1,1'-bis (dibenzyl phosphorus) dichlorodipentylpalladium, tris (dibenzylideneacetone) Dipalladium or 2-biscyclohexylphosphine-2 ', 6'-dimethoxybiphenyl, preferably [1,1'-bis (diphenylphosphino) ferrocene] palladium dichloride or 2- Dicyclohexylphosphine-2 ', 6'-dimethoxybiphenyl.

Reducing reagents include, but are not limited to: lithium aluminum hydride, sodium borohydride, DIBAL-H, NaAlH (Ot-Bu) ₃ , AlH ₃ , NaCNBH ₃ , Na (AcO) ₃ BH, BH ₃ in tetrahydrofuran solution (1N), B ₂ H ₅ , Li (Et) ₃ BH, Pd / C / H ₂ and Raney nickel / H ₂ ; Condensing agents include, but are not limited to, 1- (3-dimethylaminopropyl) -3-ethylcarbodicarbonate Imine hydrochloride, N, N'-dicyclohexylcarbodiimide, N, N'-diisopropylcarbodiimide, O-benzotriazole-N, N, N ', N'- Tetramethylurea tetrafluoroborate, 1-hydroxybenzotriazole, 1-hydroxy-7-azabenzotriazole, O-benzotriazole-N, N, N ', N'-tetramethylurea Hexafluorophosphate, 2- (7-azabenzotriazole) -N, N, N ', N'-tetramethylurea hexafluorophosphate, 2- (7-benzobenzotriazole) -N , N, N ', N' -tetramethylurea hexafluorophosphate, benzotriazol-1-yloxytris (dimethylamino) phosphonium hexafluorophosphate or hexafluorophosphate benzotriazole 1-yl-oxytripyrrolidinyl phosphorus, preferably 2- (7-azabenzotriazole) -N, N, N ', N'-tetramethylurea hexafluorophosphate; the above reaction It is preferably carried out in a solvent, including but not limited to: acetic acid, methanol, ethanol, toluene, tetrahydrofuran, dichloromethane, petroleum , Ethyl acetate, n-hexane, dimethyl sulfoxide, 1,4-dioxane, water, N, N - dimethylformamide, and mixtures thereof; wherein: R ^c is an alkyl group, preferably B X is halogen, preferably bromine; ring A, ring B, ring C, W ¹ , W ² , W ³ , R ¹ to R ³ , n, s and t are as defined in general formula (I).

The compound represented by the general formula (I) of the present invention or a tautomer, a racemate, a racemate, an enantiomer, a diastereomer or a mixture thereof, or a pharmaceutically acceptable compound thereof The preparation method of salt includes the following steps:

In the first step, the compound of the general formula (I-1) and paraformaldehyde are ring-closed under acidic conditions to obtain the general formula (I-2); in the second step, the compound of the general formula (I-2) is In the presence of a catalyst, it is reacted with CO and R ^c OH to obtain the general formula (I-3); in the third step, the compound of the general formula (I-3) is stripped of trifluoroacetamidine under basic conditions to obtain the general formula (I- 4); In the fourth step, a compound of the general formula (I-4) is reacted with a compound of the general formula (I-5) to obtain a compound of the general formula (I-7) under basic conditions in the presence of a catalyst; or A compound of the general formula (I-4) is subjected to reductive amination with a compound of the general formula (I-6) under acidic conditions in the presence of a reducing agent to obtain a compound of the general formula (I-7); in a fifth step, the general formula (I -7) The compound is hydrolyzed to obtain the (IA) compound under basic conditions. In the sixth step, the compound of the general formula (IA) and the compound of the general formula (IB) are subjected to a condensation reaction in the presence of a condensing agent under basic conditions to obtain a general compound. Compounds of formula (I); reagents that provide basic conditions include organic bases and inorganic bases, which include, but are not limited to, triethylamine, N, N-diisopropylethylamine, n-butyllithium, diiso Propylamino lithium, potassium acetate, tertiary Sodium alkoxide or potassium third butoxide, the inorganic bases include but are not limited to sodium hydride, sodium hydroxide, potassium phosphate, sodium carbonate, sodium bicarbonate, potassium carbonate or cesium carbonate; reagents providing acidic conditions include, but are not limited to Hydrogen chloride, 1,4-dioxane solution of hydrogen chloride, trifluoroacetic acid, formic acid, acetic acid, hydrochloric acid, sulfuric acid, methanesulfonic acid, nitric acid, phosphoric acid, p-toluenesulfonic acid, Me ₃ SiCl, and TMSOT _f ; catalysts include, but not Limited to palladium / carbon, Raney nickel, tetra-triphenylphosphine palladium, palladium dichloride, palladium acetate, 2-dicyclohexylphosphine-2,4,6-triisopropylbiphenyl, [1,1 ' -Bis (diphenylphosphino) ferrocene] palladium dichloride, 1,1'-bis (dibenzylphosphine) dichlorodipentylpalladium, tris (dibenzylideneacetone) dipalladium or 2- Dicyclohexylphosphine-2 ', 6'-dimethoxybiphenyl, preferably [1,1'-bis (diphenylphosphino) ferrocene] palladium dichloride or 2-dicyclohexylphosphine-2 ', 6'-Dimethoxybiphenyl.

Reducing reagents include, but are not limited to: lithium aluminum hydride, sodium borohydride, DIBAL-H, NaAlH (Ot-Bu) ₃ , AlH ₃ , NaCNBH ₃ , Na (AcO) ₃ BH, BH ₃ in tetrahydrofuran solution (1N), B ₂ H ₅ , Li (Et) ₃ BH, Pd / C / H ₂ and Raney nickel / H ₂ ; Condensing agents include, but are not limited to, 1- (3-dimethylaminopropyl) -3-ethylcarbodicarbonate Imine hydrochloride, N, N'-dicyclohexylcarbodiimide, N, N'-diisopropylcarbodiimide, O-benzotriazole-N, N, N ', N'- Tetramethylurea tetrafluoroborate, 1-hydroxybenzotriazole, 1-hydroxy-7-azabenzotriazole, O-benzotriazole-N, N, N ', N'-tetramethylurea Hexafluorophosphate, 2- (7-azabenzotriazole) -N, N, N ', N'-tetramethylurea hexafluorophosphate, 2- (7-benzobenzotriazole) -N , N, N ', N' -tetramethylurea hexafluorophosphate, benzotriazol-1-yloxytris (dimethylamino) phosphonium hexafluorophosphate or hexafluorophosphate benzotriazole 1-yl-oxytripyrrolidinyl phosphorus, preferably 2- (7-azabenzotriazole) -N, N, N ', N'-tetramethylurea hexafluorophosphate; the above reaction It is preferably carried out in a solvent, including but not limited to: acetic acid, methanol, ethanol, toluene, tetrahydrofuran, dichloromethane, petroleum , Ethyl acetate, n-hexane, dimethyl sulfoxide, 1,4-dioxane, water, N, N - dimethylformamide, and mixtures thereof; wherein: R ^c is an alkyl group, preferably B X is halogen, preferably bromine; ring A, ring B, R ¹ to R ³ , n, s and t are as defined in general formula (I).

The compound represented by the general formula (II) of the present invention or a tautomer, a racemate, a racemate, an enantiomer, a diastereomer or a mixture thereof, or a pharmaceutically acceptable form thereof The preparation method of salt includes the following steps:

In the first step, the compound of the general formula (II-1) and paraformaldehyde are ring-closed under acidic conditions to obtain the general formula (II-2); in the second step, the compound of the general formula (II-2) is In the presence of a catalyst, it is reacted with CO and R ^c OH to obtain the general formula (II-3); in the third step, the compound of the general formula (II-3) is stripped of trifluoroacetamidine under basic conditions to obtain the general formula (II- 4); In the fourth step, a compound of the general formula (II-4) is reacted with a compound of the general formula (I-5) to obtain a compound of the general formula (II-7) under basic conditions in the presence of a catalyst; or A compound of the general formula (II-4) is subjected to reductive amination with a compound of the general formula (I-6) under acidic conditions in the presence of a reducing agent to obtain a compound of the general formula (II-7); in a fifth step, the general formula (II) -7) The compound is hydrolyzed to obtain the compound (II-A) under basic conditions; in the sixth step, the compound of the general formula (II-A) and the compound of the general formula (IB) occur under basic conditions in the presence of a condensing agent Condensation reaction to obtain compounds of general formula (II); reagents providing basic conditions include organic bases and inorganic bases, the organic bases include but are not limited to triethylamine, N, N-diisopropylethylamine, n-butyl Lithium, lithium diisopropylamino, vinegar Potassium, sodium third butoxide or potassium third butoxide, the inorganic bases include but are not limited to sodium hydride, sodium hydroxide, potassium phosphate, sodium carbonate, sodium bicarbonate, potassium carbonate or cesium carbonate; Reagents include, but are not limited to, hydrogen chloride, 1,4-dioxane solution of hydrogen chloride, trifluoroacetic acid, formic acid, acetic acid, hydrochloric acid, sulfuric acid, methanesulfonic acid, nitric acid, phosphoric acid, p-benzenesulfonic acid, Me ₃ SiCl, and TMSOT _f ; Catalysts include but are not limited to palladium / carbon, Raney nickel, tetra-triphenylphosphine palladium, palladium dichloride, palladium acetate, 2-dicyclohexylphosphine-2,4,6-triisopropylbiphenyl, [1,1'-bis (diphenylphosphino) ferrocene] palladium dichloride, 1,1'-bis (dibenzyl phosphorus) dichlorodipentylpalladium, tris (dibenzylideneacetone) Dipalladium or 2-biscyclohexylphosphine-2 ', 6'-dimethoxybiphenyl, preferably [1,1'-bis (diphenylphosphino) ferrocene] palladium dichloride or 2- Dicyclohexylphosphine-2 ', 6'-dimethoxybiphenyl; reducing reagents include, but are not limited to: lithium aluminum hydride, sodium borohydride, DIBAL-H, NaAlH (Ot-Bu) ₃ , AlH ₃ , NaCNBH ₃ , Tetrahydrofuran solution of Na (AcO) ₃ BH, BH ₃ (1N), B ₂ H ₅ , Li (Et) ₃ BH, Pd / C / H ₂ and Ra NiNi / H ₂ ; Condensing agents include, but are not limited to, 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride, N, N'-dicyclohexylcarbodiimide, N, N'-diisopropylcarbodiimide, O-benzotriazole-N, N, N ', N'-tetramethylurea tetrafluoroborate, 1-hydroxybenzotriazole, 1- Hydroxy-7-azabenzotriazole, O-benzotriazole-N, N, N ', N'-tetramethylurea hexafluorophosphate, 2- (7-azabenzotriazole) -N , N, N ', N'-tetramethylurea hexafluorophosphate, 2- (7-benzobenzotriazole) -N, N, N', N' -tetramethylurea hexafluorophosphate, benzene Benzotriazol-1-yloxytris (dimethylamino) phosphonium hexafluorophosphate or benzotriazol-1-yl-oxytripyrrolidinylphosphonium hexafluorophosphate, preferably 2- (7-azabenzotriazole) -N, N, N ', N'-tetramethylurea hexafluorophosphate; the above reaction is preferably performed in a solvent, and the solvent used includes, but is not limited to, acetic acid, methanol, Ethanol, toluene, tetrahydrofuran, dichloromethane, petroleum ether, ethyl acetate, n-hexane, dimethylsulfinium, 1,4-dioxane, water, N , N -dimethylformamide and mixtures thereof; wherein: R ^c is alkyl, preferably ethyl; X is a halogen, preferably bromine; ring A, ^{B, R 1 ~ R 3,} s and t are as formula (II) as defined above.

The compound represented by the general formula (III) of the present invention or a tautomer, a racemate, a racemate, an enantiomer, a diastereomer or a mixture thereof, or a pharmaceutically acceptable form thereof The preparation method of salt includes the following steps:

In the first step, a compound of the general formula (II-4) is reacted with a compound of the general formula (III-1) under basic conditions in the presence of a catalyst to obtain a compound of the general formula (III-2); in the second step, The compound of the general formula (III-2) is hydrolyzed to obtain the compound of the general formula (III-3) under basic conditions. In the third step, the compound of the general formula (III-3) and the compound of general formula (IB) are subjected to condensation under basic conditions. A condensation reaction occurs in the presence of an agent to obtain a compound of general formula (III); reagents that provide basic conditions include organic bases and inorganic bases, and the organic bases include, but are not limited to, triethylamine, N, N-diisopropylethylamine , N-butyllithium, lithium diisopropylamino, potassium acetate, sodium tert-butoxide or potassium tert-butoxide, the inorganic bases include, but are not limited to, sodium hydride, sodium hydroxide, potassium phosphate, sodium carbonate, Sodium bicarbonate, potassium carbonate or cesium carbonate; catalysts include, but are not limited to, palladium / carbon, Raney nickel, tetra-triphenylphosphine palladium, palladium dichloride, palladium acetate, 2-dicyclohexyl phosphorus-2,4, 6-triisopropylbiphenyl, [1,1'-bis (diphenylphosphino) ferrocene] palladium dichloride, 1,1'-bis (dibenzylphosphine) dichlorodipentylpalladium , Tris (dibenzylideneacetone) dipalladium or 2-bicyclic Hexylphosphine-2 ', 6'-dimethoxybiphenyl, preferably [1,1'-bis (diphenylphosphino) ferrocene] palladium dichloride or 2-biscyclohexylphosphine-2' , 6'-dimethoxybiphenyl; condensing agents include, but are not limited to, 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride, N, N'-dicyclohexyl Carbodiimide, N, N'-diisopropylcarbodiimide, O-benzotriazole-N, N, N ', N'-tetramethylurea tetrafluoroborate, 1-hydroxybenzo Triazole, 1-hydroxy-7-azabenzotriazole, O-benzotriazole-N, N, N ', N'-tetramethylurea hexafluorophosphate, 2- (7-azabenzo Triazole)-N, N, N ', N'-tetramethylurea hexafluorophosphate, 2- (7-benzotriazole oxide) -N, N, N', N ' -tetramethylurea Hexafluorophosphate, benzotriazol-1-yloxytris (dimethylamino) phosphonium hexafluorophosphate, or benzotriazol-1-yl-oxytripyrrolidinylphosphonium hexafluorophosphate , Preferably 2- (7-azabenzotriazole) -N, N, N ', N'-tetramethylurea hexafluorophosphate; the above reaction is preferably performed in a solvent, and the solvent used includes but not Limited to: acetic acid, methanol, ethanol, toluene, tetrahydrofuran, dichloromethane, petroleum ether, ethyl acetate, n-hexane, dimethylsulfine, 1,4-dioxane Water, N, N - dimethylformamide, and mixtures thereof; wherein: R ^c is alkyl, preferably ethyl; X is a halogen, preferably bromine; ring ^{B, G 1, G 2,} R 1 ~ R ³ and t are as defined in general formula (III).

The present invention is further described below with reference to examples, but these examples do not limit the scope of the present invention.

    Examples    

The structure of the compound is determined by nuclear magnetic resonance (NMR) or / and mass spectrometry (MS). The NMR shift (δ) is given in units of 10 ^-6 (ppm). The NMR measurement was performed using Bruker AVANCE-400 nuclear magnetometer. The measurement solvents were deuterated dimethylsulfinium (DMSO- d ₆ ), deuterated chloroform (CDCl ₃ ), and deuterated methanol (CD ₃ OD). The internal standard was four. Methylsilane (TMS).

MS was measured using a FINNIGAN LCQAd (ESI) mass spectrometer (manufacturer: Thermo, model: Finnigan LCQ advantage MAX).

The HPLC was measured using an Agilent 1200 DAD high-pressure liquid chromatography (Sunfire C18 150 × 4.6 mm column) and a Waters 2695-2996 high-pressure liquid chromatography (Gimini C18 150 × 4.6 mm column).

Chiral HPLC analysis was performed using LC-10A vp (Shimadzu) or SFC-analytical (Berger Instruments Inc.).

The thin-layer chromatography silica gel plate uses Yantai Huanghai HSGF254 or Qingdao GF254 silicone plate. The thin-layer chromatography (TLC) silicon plate uses a size of 0.15mm ~ 0.2mm. The thin-layer chromatography separation and purification product uses a size of 0.4mm. ~ 0.5mm.

Column chromatography generally uses Yantai Huanghai Silicone 200 ~ 300 mesh silica gel as the carrier.

Chiral preparative column chromatography used Prep Star SD-1 (Varian Instruments Inc.) or SFC-multigram (Berger Instruments Inc.).

The average inhibition rate of the kinase and the IC ₅₀ value were measured using a NovoStar microplate reader (BMG, Germany).

The known starting materials of the present invention can be synthesized by or in accordance with methods known in the art, or can be purchased from ABCR GmbH & Co. KG, Acros Organics, Aldrich Chemical Company, Accela ChemBio Inc, Dari Chemicals and other companies.

There is no special description in the examples, and the reaction can be performed under an argon atmosphere or a nitrogen atmosphere.

An argon or nitrogen atmosphere means that the reaction flask is connected to an argon or nitrogen balloon with a volume of about 1 L.

The hydrogen atmosphere means that a reaction balloon is connected to a hydrogen gas balloon with a volume of about 1 L.

Pressurized hydrogenation reaction uses Parr 3916EKX type hydrogenation instrument and clear blue QL-500 type hydrogen generator or HC2-SS type hydrogenation instrument.

The hydrogenation reaction is usually evacuated and filled with hydrogen, and the operation is repeated 3 times.

For the microwave reaction, a CEM Discover-S 908860 microwave reactor was used.

There is no special description in the examples, and the solution means an aqueous solution.

There is no special description in the examples, and the reaction temperature is room temperature, which is 20 ° C to 30 ° C.

The monitoring of the reaction progress in the examples uses thin layer chromatography (TLC), a developing agent used in the reaction, a column chromatography eluent system for purifying compounds, and a thin layer chromatography developing system including: A : Dichloromethane / methanol system, B: n-hexane / ethyl acetate system, C: petroleum ether / ethyl acetate system, D: acetone, E: dichloromethane / acetone system, F: ethyl acetate / dichloromethane System, G: ethyl acetate / dichloromethane / n-hexane, H: ethyl acetate / dichloromethane / acetone, the volume ratio of the solvent is adjusted according to the polarity of the compound, and a small amount of triethylamine and acetic acid can also be added Wait for alkaline or acidic reagents to adjust.

    Example 1     N- (4- (ethylsulfonyl) benzyl) -2 '-(4- (trifluoromethyl) phenyl) -2', 3'-dihydro-1 ' H -spiro [cyclopropane-1 , 4'-isoquinoline] -7'-formamidine 1

    First step     1- (7'-bromo-1 ' H -spiro [cyclopropane-1,4'-isoquinoline] -2' (3 ' H ) -yl) -2,2,2-trifluoroethanone 1b

N -((1- (4-bromophenyl) cyclopropyl) methyl) -2,2,2-trifluoroacetamidamine 1a (22 g, 68.3 mmol, using the method disclosed in PCT patent application "WO2011124093" Prepared) was dissolved in 150 mL of a pre-mixed solvent of acetic acid and sulfuric acid (V / V = 2: 3), paraformaldehyde (7.96 g, 264.99 mmol) was added, and the reaction was stirred for 12 hours. The reaction solution was poured into 500 mL of ice water and extracted with ethyl acetate (500 mL × 2). The organic phases were combined, washed with water, saturated sodium bicarbonate solution and saturated sodium chloride solution in that order, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude title compound 1b (4.5 g) was obtained, and the product was directly subjected to the next reaction without purification.

    Second step     2 '- (2,2,2-trifluoro-acetyl) -2', 3'-dihydro -1 'H - spiro [cyclopropane-1,4'-isoquinolin] -7'-carboxylic acid Ethyl 1c

The crude compound 1b (4 g, 11.97 mmol) was dissolved in 40 mL of a mixed solvent of dimethylarsine (V / V = 1: 1), and 1,3-bis (diphenylphosphine) propane (987.5 mg, 2.39 mmol), triethylamine (1.21 g, 11.97 mmol) and palladium acetate (537.53 mg, 2.39 mmol). Under a carbon monoxide atmosphere, the temperature was raised to 60 ° C. and the reaction was stirred for 12 hours. The reaction solution was cooled to room temperature, poured into 100 mL of water, and extracted with ethyl acetate (100 mL × 2). The organic phases were combined, washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by column chromatography with eluent system B to obtain the title compound 1c (2.5 g, yield: 63.8%).

    third step     2 ', 3'-dihydro -1' H - spiro [cyclopropane-1,4'-isoquinolin] -7'-carboxylate 1d

1c (2.5 g, 7.64 mmol) was dissolved in 100 mL of a mixed solvent of ethanol and water (V / V = 1: 1), potassium carbonate (1.58 g, 11.46 mmol) was added, and the reaction was stirred at room temperature for 12 hours. The reaction solution was poured into 100 mL of water and extracted with ethyl acetate (100 mL × 2). The organic phases were combined, washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give the crude title compound 1d (2g). The product was directly subjected to the next reaction without purification.

    the fourth step     2 '- (4- (trifluoromethyl) phenyl) -2', 3'-dihydro -1 'H - spiro [cyclopropane-1,4'-isoquinolin] -7'-acetic acid Ester 1f

The crude compound 1d (2g, 8.65mmol) and 1-bromo-4- (trifluoromethyl) benzene 1e (2.33g, 10.38mmol) were prepared by a known method " Organic Letters , 2014, 16 (16), 4268-4271 "Prepared" was dissolved in 50mL of 1,4-dioxane, and cesium carbonate (8.45g, 25.94mmol), 2-dicyclohexylphosphine-2,4,6-triisopropylbiphenyl (412.22mg, 864.72umol) and tris (dibenzylideneacetone) dipalladium (791.83mg, 864.72umol), the temperature was raised to 90 ° C and the reaction was stirred for 3 hours. The reaction solution was cooled to room temperature, poured into 100 mL of water, and extracted with ethyl acetate (100 mL × 2). The organic phases were combined, washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by column chromatography with eluent system B to obtain the title compound 1f (1.7 g, yield: 52.37%).

    the fifth step     2 '- (4- (trifluoromethyl) phenyl) -2', 3'-dihydro -1 'H - spiro [cyclopropane-1,4'-isoquinolin] -7'-carboxylate 1g

Compound 1f (1.5 g, 4 mmol) was dissolved in 60 mL of a mixed solvent of ethanol and water (V / V = 5: 1), sodium hydroxide (799.12 mg, 19.98 mmol) was added, and the mixture was heated to reflux and stirred for 2 hours. The reaction solution was cooled to room temperature, and concentrated under reduced pressure. The resulting residue was added dropwise with 1 M hydrochloric acid to pH <2, and extracted with ethyl acetate (50 mL × 2). The organic phases were combined, washed with a saturated sodium chloride solution, and anhydrous sulfuric acid. Sodium was dried, filtered, and the filtrate was concentrated under reduced pressure to obtain 1 g (1.5 g) of the crude title compound. The product was directly subjected to the next reaction without purification.

    Step Six     N- (4- (ethylsulfonyl) benzyl) -2 '-(4- (trifluoromethyl) phenyl) -2', 3'-dihydro-1 ' H -spiro [cyclopropane-1 , 4'-isoquinoline] -7'-formamidine 1

Crude compound 1 g (1.4 g, 4.03 mmol) and (4- (ethylsulfonyl) phenyl) methylamine for 1 h (1.2 g, 6.05 mmol, prepared by the method disclosed in PCT patent application "WO2015017335") were dissolved in To 30mL of N , N -dimethylformamide, 2- (7-azabenzotriazole) -N, N, N ,, N'-tetramethylurea hexafluorophosphate (3.06g, 8.06 mmol) and triethylamine (1.22 g, 12.09 mmol), and the reaction was stirred for 12 hours. The reaction solution was poured into 100 mL of water and extracted with ethyl acetate (100 mL × 2). The organic phases were combined, washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained product was purified by high performance liquid chromatography. The residue was obtained as the title compound 1 (1.1 g, yield: 51.63%).

MS m / z (ESI): 529.5 [M + 1]

¹ H NMR (400MHz, DMSO- d ₆ ) δ 9.11 (s, 1H), 7.80-7.85 (m, 3H), 7.69-7.71 (m, 1H), 7.55-7.57 (m, 2H), 7.49-7.50 ( m, 2H), 7.11-7.13 (m, 2H), 6.93-6.95 (m, 1H), 4.65 (s, 2H), 4.56-4.57 (m, 2H), 3.54 (s, 2H), 3.23-3.28 ( m, 2H), 1.05-1.09 (m, 7H).

    Example 2     N- (1-4- (ethylsulfonyl) phenyl) -2-hydroxyethyl) -2 '-(4- (trifluoromethyl) phenyl) -2', 3'-dihydro-1 ' H -spiro [cyclopropane-1,4'-isoquinoline] -7'-formamidine 2

Crude compound 1g (200mg, 575.82umol) and 2-amino-2- (4- (ethylsulfonyl) phenyl) ethanol 2a (198.05mg, 863.73umol) were prepared by the method disclosed in PCT patent application "WO2016061160" And obtained) dissolved in 10 mL of N , N -dimethylformamide, and added 2- (7-azabenzotriazole) -N, N, N ', N'-tetramethylurea hexafluorophosphate (328.22 mg, 863.73 umol) and triethylamine (174.8 mg, 1.73 mmol). The reaction was stirred for 12 hours. The reaction solution was poured into 100 mL of water and extracted with ethyl acetate (100 mL × 2). The organic phases were combined, washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained product was purified by high performance liquid chromatography. The residue was obtained as the title compound 2 (120 mg, yield: 37.31%).

MS m / z (ESI): 559.1 [M + 1]

¹ H NMR (400MHz, DMSO- d ₆ ) δ 8.75-8.77 (m, 1H), 7.80-7.84 (m, 3H), 7.70-7.72 (m, 1H), 7.64-7.66 (m, 2H), 7.49- 7.51 (m, 2H), 7.12-7.14 (m, 2H), 6.92-6.95 (m, 1H), 5.11-5.17 (m, 1H), 4.66 (s, 2H), 4.23 (s, 1H), 3.69- 3.75 (m, 2H), 3.54 (s, 2H), 3.24-3.29 (m, 2H), 1.03-1.11 (m, 7H).

    Examples 3, 4     (R) - N - (1-4- ( ethanesulfonamide acyl) phenyl) -2-hydroxyethyl) -2 '- (4- (trifluoromethyl) phenyl) -2', 3'- Dihydro-1 ' H -spiro [cyclopropane-1,4'-isoquinoline] -7'-formamidine 3 (S) - N - (1-4- ( ethanesulfonamide acyl) phenyl) -2-hydroxyethyl) -2 '- (4- (trifluoromethyl) phenyl) -2', 3'- Dihydro-1 ' H -spiro [cyclopropane-1,4'-isoquinoline] -7'-formamidine 4

Chiral preparation of 2 (120 mg, 0.215 mmol) (separation conditions: chiral preparation column Lux Cellulose-1 OD 4.6 * 150mm 5 μm (with guard column); mobile phase: n-hexane: ethanol: trifluoroacetic acid = 50: 50: 0.01, flow rate: 1.0 mL / min), the corresponding components were collected, concentrated under reduced pressure, and the resulting residue was purified by high performance liquid chromatography to obtain the title compounds 3 (10 mg) and 4 (20 mg).

Compound 3:

MS m / z (ESI): 559.0 [M + 1]; chiral HPLC analysis: retention time 4.411 minutes, chiral purity: 93% (chromatographic column: Lux Cellulose-1 OD 4.6 * 150mm 5um (with protective column) )); Mobile phase: n-hexane / ethanol / trifluoroacetic acid = 50/50 / 0.01 (v / v / v)).

¹ H NMR (400MHz, DMSO- d ₆ ) δ 8.74-8.76 (m, 1H), 7.79-7.84 (m, 3H), 7.64-7.72 (m, 3H), 7.49-7.51 (m, 2H), 7.12- 7.14 (m, 2H), 6.92-6.95 (m, 1H), 5.13-5.15 (m, 1H), 4.66 (s, 2H), 3.70-3.75 (s, 2H), 3.54 (s, 2H), 3.24- 3.29 (m, 2H), 0.85-1.11 (m, 7H).

Compound 4:

MS m / z (ESI): 559.1 [M + 1]; chiral HPLC analysis: retention time 6.9148 minutes, chiral purity: 90%. % (Chromatographic column: Superchiral S-AD (Chiralway), 0.46 cm I.D. * 25 cm, 5 um; mobile phase: n-hexane / ethanol / trifluoroacetic acid = 50/50 / 0.01 (v / v / v)).

¹ H NMR (400MHz, DMSO- d ₆ ) δ 8.74-8.75 (m, 1H), 7.79-7.84 (m, 3H), 7.64-7.71 (m, 3H), 7.49-7.51 (m, 2H), 7.12- 7.14 (m, 2H), 6.92-6.94 (m, 1H), 5.13-5.15 (m, 1H), 4.66 (s, 2H), 3.67-3.77 (m, 2H), 3.54 (s, 2H), 3.23- 3.29 (m, 2H), 0.84-1.11 (m, 7H).

    Example 5     N -((5- (ethylsulfonyl) pyridin-2-yl) methyl) -2 '-(4- (trifluoromethyl) phenyl) -2', 3'-dihydro-1 ' H -Spiro [cyclopropane-1,4'-isoquinoline] -7'-formamidine

1 g (50 mg, 0.14 mmol) of crude compound and 5- (ethylsulfonyl) pyridin-2-yl) methylamine 5a (43.24 mg, 0.22 mmol, prepared by the method disclosed in PCT patent application "WO2015017335") were dissolved. In 10 mL of N , N -dimethylformamide, 2- (7-azabenzotriazole) -N, N, N ', N'-tetramethylurea hexafluorophosphate (109.41mg, 0.29 mmol) and triethylamine (43.62 mg, 0.43 mmol), and the reaction was stirred for 12 hours. The reaction solution was poured into 30 mL of water and extracted with ethyl acetate (30 mL × 2). The organic phases were combined, washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained product was purified by high performance liquid chromatography. The residue was obtained as the title compound 5 (20 mg, yield: 24.92%).

MS m / z (ESI): 530.1 [M + 1]

¹ H NMR (400MHz, DMSO- d ₆ ) δ 9.22 (s, 1H), 8.95-8.96 (m, 1H), 8.24-8.26 (m, 1H), 7.83 (s, 1H), 7.74-7.75 (m, 1H), 7.57-7.59 (m, 1H), 7.50-7.52 (m, 2H), 7.13-7.15 (m, 2H), 6.95-6.97 (m, 1H), 4.67 (s, 4H), 3.67-3.70 ( m, 2H), 3.37-3.40 (m, 2H), 1.07-1.14 (m, 7H).

    Example 6     N -((1- (ethanesulfonyl) piperidin-4-yl) methyl) -2 '-(4- (trifluoromethyl) phenyl) -2', 3'-dihydro-1 ' H -spiro [cyclopropane-1,4'-isoquinoline] -7'-formamide 6

1 g (100 mg, 0.287 mmol) of crude compound and (1- (ethanesulfonyl) piperidin-4-yl) methylamine 6a (89.1 mg, 0.432 mmol) were prepared by the method disclosed in the US patent application "US20160122318" ) Dissolved in 20mL N , N -dimethylformamide, and added 2- (7-azabenzotriazole) -N, N, N ', N'-tetramethylurea hexafluorophosphate (164.11 mg, 0.432 mmol) and triethylamine (87.4 mg, 0.864 mmol), and the reaction was stirred for 3 hours. The reaction solution was poured into 50 mL of water and extracted with ethyl acetate (50 mL × 2). The organic phases were combined, washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained product was purified by high performance liquid chromatography. The residue was obtained as the title compound 6 (50 mg, yield: 34.42%).

MS m / z (ESI): 536.5 [M + 1]

¹ H NMR (400MHz, DMSO- d ₆ ) δ 8.46 (s, 1H), 7.74 (s, 1H), 7.63-7.65 (m, 1H), 7.49-7.52 (m, 2H), 7.12-7.14 (m, 2H), 6.90-6.92 (m, 1H), 4.65 (s, 2H), 3.53-3.56 (m, 3H), 3.16-3.19 (m, 3H), 2.99-3.05 (m, 2H), 2.73-2.78 ( m, 2H), 1.73-1.76 (m, 3H), 1.02-1.23 (m, 9H).

    Example 7     N- (4- (ethanesulfonyl) benzyl) -2 '-((1 r , 4 r ) -4- (trifluoromethyl) cyclohexyl) -2', 3'-dihydro-1 ' H -spiro [cyclopropane-1,4'-isoquinoline] -7'-formamide 7

    First step     2 '- (4- (trifluoromethyl) cyclohexyl) -2', 3'-dihydro -1 'H - spiro [cyclopropane-1,4'-isoquinolin] -7'-acetic acid Ester 7b

The crude compound 1d (300 mg, 1, 3 mmol) was dissolved in 10 mL of methanol, and acetic acid (15.58 mg, 0.259 mmol) and sodium triacetoxyborohydride (549.81 mg, 2.59 mmol) were added, and the reaction was stirred for 2 hours. 4- (trifluoromethyl) cyclohexane-1-one 7a (430.99 mg, 2.59 mmol, prepared by the method disclosed in PCT patent application "WO2008007930"), and stirred for 12 hours. The reaction solution was poured into 50 mL of water and extracted with ethyl acetate (50 mL × 2). The organic phases were combined, washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, and washed with silica gel column chromatography. The obtained residue was purified by using a solvent removing system B to obtain the title compound 7b (100 mg, yield: 20.21%).

    Second step     2 '- (4- (trifluoromethyl) cyclohexyl) -2', 3'-dihydro -1 'H - spiro [cyclopropane-1,4'-isoquinolin] -7' acid 7c

Compound 7b (100 mg, 0.262 mmol) was dissolved in 13 mL of a mixed solvent of ethanol and water (V / V = 10: 3), sodium hydroxide (52.43 mg, 1.31 mmol) was added, and the mixture was heated to 70 ° C. and stirred for 2 hours. The reaction solution was cooled to room temperature, and concentrated under reduced pressure. The resulting residue was added dropwise 1 M hydrochloric acid to pH <2, and extracted with ethyl acetate (30 mL × 2). The organic phases were combined, washed with a saturated sodium chloride solution, and anhydrous sulfuric acid. Sodium was dried, filtered, and the filtrate was concentrated under reduced pressure to give the crude title compound 7c (92 mg). The product was directly subjected to the next reaction without purification.

    third step     N- (4- (ethanesulfonyl) benzyl) -2 '-((1 r , 4 r ) -4- (trifluoromethyl) cyclohexyl) -2', 3'-dihydro-1 ' H -spiro [cyclopropane-1,4'-isoquinoline] -7'-formamide 7

The crude compound 7c (50 mg, 0.141 mmol) and compound 1h (33.83 mg, 0.17 mmol) were dissolved in 5 mL of N , N -dimethylformamide, and 2- (7-azabenzotriazole) -N was added. , N, N ', N'-tetramethylurea hexafluorophosphate (80.65 mg, 0.212 mmol) and triethylamine (42.95 mg, 0.424 mmol), and the reaction was stirred for 12 hours. The reaction solution was poured into 50 mL of water and extracted with ethyl acetate (50 mL × 2). The organic phases were combined, washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained product was purified by high performance liquid chromatography. The residue gave the title compound 7 (20 mg, yield: 26.5%).

MS m / z (ESI): 535.2 [M + 1]

¹ H NMR (400MHz, DMSO- d ₆ ) δ 9.03 (s, 1H), 7.83-7.85 (m, 2H), 7.55-7.64 (m, 4H), 6.79-6.81 (m, 1H), 4.55-4.57 ( m, 2H), 3.84 (s, 2H), 3.23-3.27 (m, 2H), 2.61 (s, 2H), 1.94 (s, 3H), 1.24-1.36 (m, 7H), 0.90-1.11 (m, 7H).

    Example 8     N- (4- (ethanesulfonyl) benzyl) -2 '-(2- (trifluoromethyl) phenyl) -2', 3'-dihydro-1 ' H -spiro [cyclopropane-1 , 4'-isoquinoline] -7'-formamidine 8

    First step     2 '- (2- (trifluoromethyl) phenyl) -2', 3'-dihydro -1 'H - spiro [cyclopropane-1,4'-isoquinolin] -7'-acetic acid Ester 8b

The crude compound 1d (200 mg, 0.864 mmol) and 1-bromo-2- (trifluoromethyl) benzene 8a (233.48 mg, 1.04 mmol, using a well-known method " European Journal , 2013, 19 (52), 17692-17697 "Prepared" was dissolved in 3 mL of 1,4-dioxane, and sodium tert-butoxide (249.31 mg, 2.59 mmol), 2-dicyclohexyl phosphorus-2,4,6-triisopropylbiphenyl ( 82.44 mg, 172.94 umol) and tris (dibenzylideneacetone) dipalladium (158.37 mg, 172.94 umol). The temperature was raised to 105 ° C and the reaction was stirred for 18 hours. The reaction solution was cooled to room temperature, poured into 10 mL of water, and extracted with ethyl acetate (20 mL × 2). The organic phases were combined, washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by column chromatography with eluent System B to obtain the title compound 8b (130 mg, yield: 40.05%).

    Second step     2 '- (2- (trifluoromethyl) phenyl) -2', 3'-dihydro -1 'H - spiro [cyclopropane-1,4'-isoquinolin] -7'-carboxylic acid 8c

Compound 8b (130 mg, 0.346 mmol) was dissolved in 12 mL of a mixed solvent of ethanol and water (V / V = 5: 1), sodium hydroxide (138.51 mg, 3.46 mmol) was added, and the mixture was heated to 60 ° C. and stirred for 2 hours. The reaction solution was cooled to room temperature, and concentrated under reduced pressure. 1 M hydrochloric acid was added dropwise to the resulting residue to a pH of 3 to 4, and the mixture was concentrated under reduced pressure to obtain the crude title compound 8c (120 mg). The product was directly subjected to the next reaction without purification.

    third step     N- (4- (ethanesulfonyl) benzyl) -2 '-(2- (trifluoromethyl) phenyl) -2', 3'-dihydro-1 ' H -spiro [cyclopropane-1 , 4'-isoquinoline] -7'-formamidine 8

The crude compound 8c (100 mg, 0.288 mmol) and compound 1h (114.74 mg, 0.576 mmol) were dissolved in 2 mL of N , N -dimethylformamide, and 2- (7-azabenzotriazole) -N was added. , N, N ', N'-tetramethylurea hexafluorophosphate (218 mg, 0.576 mmol) and N , N -diisopropylethylamine (111.63 mg, 0.864 mmol), and the reaction was stirred for 1 hour. The reaction solution was added with 10 mL of water and extracted with ethyl acetate (10 mL × 2). The organic phases were combined, washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by high performance liquid chromatography. The title compound 8 was obtained (40 mg, yield: 26.28%).

MS m / z (ESI): 529.5 [M + 1]

¹ H NMR (400MHz, CDCl3) δ 7.85-7.87 (m, 2H), 7.66-7.68 (m, 1H), 7.59-7.61 (m, 1H), 7.52-7.54 (m, 4H), 7.38-7.40 (m , 1H), 6.80-6.82 (m, 1H), 6.60-6.63 (m, 1H), 4.74-4.75 (m, 2H), 4.29 (s, 2H), 3.08-3.13 (m, 4H), 1.25-1.29 (m, 3H), 1.07-1.10 (m, 2H), 0.95-0.98 (m, 2H).

    Example 9     N - (4- (ethanesulfonyl acyl) benzyl) -2 '- (5- (trifluoromethyl) pyrimidin-2-yl) -2', 3'-dihydro -1 'H - spiro [cyclopropane Propane-1,4'-isoquinoline] -7'-formamide 9

    First step     2 '-(5- (trifluoromethyl) pyrimidin-2-yl) -2', 3'-dihydro-1 ' H -spiro [cyclopropane-1,4'-isoquinoline] -7'- Ethyl carboxylate 9b

The crude compound 1d (100 mg, 0.432 mmol) and 2-chloro-5- (trifluoromethyl) pyrimidine 9a (118 mg, 0.646 mmol) were dissolved in 2 mL of DMSO, and 1,8-diazabicyclo [5.4.0 ] Undecyl-7-ene (326 mg, 1.294 mmol), and the temperature was raised to 90 ° C. and the reaction was stirred for 18 hours. The reaction solution was cooled to room temperature, poured into 10 mL of water, and extracted with ethyl acetate (20 mL × 2). The organic phases were combined, washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by column chromatography with eluent System B to obtain the title compound 9b (100 mg, yield: 61.29%).

    Second step     2 '-(5- (trifluoromethyl) pyrimidin-2-yl) -2', 3'-dihydro-1 ' H -spiro [cyclopropane-1,4'-isoquinoline] -7'- Carboxylic acid 9c

Compound 9b (50 mg, 0.132 mmol) was dissolved in 12 mL of a mixed solvent of ethanol and water (V / V = 5: 1), sodium hydroxide (53 mg, 1.32 mmol) was added, and the temperature was raised to 60 ° C. and the reaction was stirred for 2 hours. The reaction solution was cooled to room temperature, and concentrated under reduced pressure. 1 M hydrochloric acid was added dropwise to the resulting residue to a pH of 3 to 4, and concentrated under reduced pressure to obtain the crude title compound 9c (50 mg). The product was directly subjected to the next reaction without purification.

    third step     N - (4- (ethanesulfonyl acyl) benzyl) -2 '- (5- (trifluoromethyl) pyrimidin-2-yl) -2', 3'-dihydro -1 'H - spiro [cyclopropane Propane-1,4'-isoquinoline] -7'-formamide 9

The crude compound 9c (50mg, 0.143mmol) and compound 1h (28mg, 0.140mmol) were dissolved in 2mL of N , N -dimethylformamide, and 2- (7-azabenzotriazole) -N was added, N, N ', N'-tetramethylurea hexafluorophosphate (82 mg, 0.216 mmol) and N , N -diisopropylethylamine (37 mg, 0.286 mmol) were stirred for 1 hour. The reaction solution was added with 10 mL of water and extracted with ethyl acetate (10 mL × 2). The organic phases were combined, washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by high performance liquid chromatography. The title compound 9 was obtained (50 mg, yield: 65.84%).

MS m / z (ESI): 531.4 [M + 1]

¹ H NMR (400MHz, CDCl ₃ ) δ 8.49-8.51 (m, 2H), 7.65-7.75 (m, 4H), 7.44-7.46 (m, 2H), 7.03-7.06 (m, 1H), 6.85-6.87 ( m, 1H), 5.13-5.16 (m, 2H), 4.71-4.73 (m, 2H), 3.96-3.98 (m, 2H), 3.06-3.11 (m, 2H), 1.24-1.27 (m, 3H), 1.07-1.12 (m, 4H).

    Example 10     N - (4- (ethanesulfonyl acyl) benzyl) -2 '- (4- (trifluoromethyl) phenyl) -2', 3'-dihydro -1 'H - spiro [cyclobutane - 1,4'-isoquinoline] -7'-formamidine 10

    First step     1- (7'-bromo-1 ' H -spiro [cyclobutane-1,4'-isoquinoline] -2' (3 ' H ) -yl) -2,2,2-trifluoroethanone 10b

N -((1- (4-bromophenyl) cyclobutyl) methyl) -2,2,2-trifluoroacetamidamine 10a (1.2 g, 3.57 mmol, using the method disclosed in the patent application "WO2011124093" Prepared) was dissolved in 27 mL of a mixed solvent of pre-made acetic acid and sulfuric acid (V / V = 2: 3), paraformaldehyde (321 mg, 10.69 mmol) was added, and the reaction was stirred for 12 hours. The reaction solution was poured into 100 mL of ice water and extracted with ethyl acetate (50 mL × 2). The organic phases were combined, washed with water, saturated sodium bicarbonate solution and saturated sodium chloride solution in that order, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude title compound 10b (1 g) was obtained, and the product was directly subjected to the next reaction without purification.

    Second step     2 '- (2,2,2-trifluoro-acetyl) -2', 3'-dihydro -1 'H - spiro [cyclobutane-1,4'-isoquinolin] -7'-carboxamide Ethyl Ester 10c

The crude compound 10b (1 g, 2.87 mmol) was dissolved in 20 mL of a mixed solvent of dimethylarsine (V / V = 1: 1), and 1,3-bis (diphenylphosphine) propane (118 mg, 0.28) was added. mmol), triethylamine (290 mg, 2.86 mmol) and palladium acetate (64 mg, 0.28 mmol). Under a carbon monoxide atmosphere, the temperature was raised to 60 ° C. and the reaction was stirred for 12 hours. The reaction solution was cooled to room temperature, poured into 100 mL of water, and extracted with ethyl acetate (100 mL × 2). The organic phases were combined, washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by column chromatography with eluent system B to obtain the title compound 10c (400 mg, yield: 40.8%).

    third step     2 ', 3'-dihydro -1' H - spiro [cyclobutane-1,4'-isoquinolin] -7'-carboxylate 10d

10c (400 mg, 1.17 mmol) was dissolved in 50 mL of a mixed solvent of ethanol and water (V / V = 1: 1), potassium carbonate (485 mg, 3.51 mmol) was added, and the reaction was stirred at room temperature for 12 hours. The reaction solution was poured into 100 mL of water and extracted with ethyl acetate (50 mL × 2). The organic phases were combined, washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude title compound 10d (350 mg). The product was directly subjected to the next reaction without purification.

    the fourth step     2 '- (4- (trifluoromethyl) phenyl) -2', 3'-dihydro -1 'H - spiro [cyclobutane-1,4'-isoquinolin] -7'-carboxylic acid Ethyl 10f

The crude compound 10d (350 mg, 1.42 mmol) and 1-bromo-4- (trifluoromethyl) benzene 10e (481 mg, 2.13 mmol) were prepared by a known method " Organic Letters , 2014, 16 (16), 4268-4271" (Prepared) was dissolved in 30 mL of 1,4-dioxane, and cesium carbonate (1.4 g, 4.29 mmol) and 2-dicyclohexylphosphine-2,4,6-triisopropylbiphenyl (68 mg, 142.64 μmol) were added. ) And tris (dibenzylideneacetone) dipalladium (130 mg, 141.96 μmol), and the reaction was stirred for 3 hours while being heated to 90 ° C. under the protection of helium. The reaction solution was cooled to room temperature, poured into 50 mL of water, and extracted with ethyl acetate (50 mL × 2). The organic phases were combined, washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by column chromatography with eluent system B to obtain the title compound 10f (400 mg, yield: 71.99%).

    the fifth step     2 '- (4- (trifluoromethyl) phenyl) -2', 3'-dihydro -1 'H - spiro [cyclobutane-1,4'-isoquinolin] -7'-carboxylic acid 10g

Compound 10f (400 mg, 1.03 mmol) was dissolved in 30 mL of a mixed solvent of ethanol and water (V / V = 5: 1), sodium hydroxide (164 mg, 4.10 mmol) was added, and the mixture was heated to reflux and stirred for 2 hours. The reaction solution was cooled to room temperature, and concentrated under reduced pressure. The resulting residue was added dropwise with 1 M hydrochloric acid to pH <2, and extracted with ethyl acetate (50 mL × 2). The organic phases were combined, washed with a saturated sodium chloride solution, and anhydrous sulfuric acid. Sodium was dried, filtered, and the filtrate was concentrated under reduced pressure to obtain 10 g (300 mg) of the crude title compound. The product was directly subjected to the next reaction without purification.

    Step Six     N - (4- (ethanesulfonyl acyl) benzyl) -2 '- (4- (trifluoromethyl) phenyl) -2', 3'-dihydro -1 'H - spiro [cyclobutane - 1,4'-isoquinoline] -7'-formamidine 10

10 g (300 mg, 0.83 mmol) of the crude compound and (4- (ethylsulfonyl) phenyl) methylamine for 10 h (198 mg, 0.99 mmol, prepared by the method disclosed in the patent application "WO2015017335") were dissolved in 10 mL of N , To N -dimethylformamide, 2- (7-azabenzotriazole) -N, N, N ', N'-tetramethylurea hexafluorophosphate (630 mg, 1.66 mmol) and tris Ethylamine (252 mg, 2.49 mmol) was stirred for 12 hours. The reaction solution was poured into 50 mL of water and extracted with ethyl acetate (50 mL × 2). The organic phases were combined, washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained product was purified by high performance liquid chromatography. The residue gave the title compound 10 (200 mg, yield: 44.39%).

MS m / z (ESI): 543.2 [M + 1]

¹ H NMR (400MHz, DMSO- d ₆ ) δ 9.13-9.15 (m, 1H), 7.83-7.85 (m, 3H), 7.74-7.79 (m, 2H), 7.53-7.58 (m, 4H), 7.16- 7.18 (m, 2H), 4.58-4.59 (m, 2H), 4.49 (s, 2H), 3.67 (s, 2H), 3.23-3.28 (m, 2H), 2.22-2.35 (m, 3H), 2.01- 2.10 (m, 3H), 1.06-1.09 (m, 3H).

    Example 11     N- (4- (ethylsulfonyl) benzyl) -2- (4- (trifluoromethyl) phenyl) -2,2 ', 3,3', 5 ', 6'-hexahydro-1 H -spiro [isoquinoline-4,4'-pyran] -7-formamidine 11

    First step     4- (4-bromophenyl) tetrahydro-2 H -pyran-4-carbonitrile 11b

Dissolve 2- (4-bromophenyl) acetonitrile 11a (2.5 g, 12.75 mmol) in 30 mL of dimethylarsine, lower the temperature to 0 ° C, slowly add sodium hydrogen (1.02 g, 25.50 mmol), and stir for 30 minutes. Add 1-bromo-2- (2-bromoethoxy) ethane (3.22 g, 14.03 mmol) and stir at room temperature for 12 hours. The reaction solution was poured into 100 mL of ice water, and extracted with ethyl acetate (50 mL × 2). The organic phases were combined, washed with water, saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained residue was purified by the eluent system B to obtain the title compound 11b (2.5 g, yield: 73.75%).

    Second step     (4- (4-Bromophenyl) tetrahydro-2 H -pyran-4-yl) methylamine 11c

The crude compound 11b (2.5 g, 9.39 mmol) was dissolved in 100 mL of tetrahydrofuran, and a 1M borane tetrahydrofuran solution (28.18 ml, 28.18 mmol) was added. The reaction was heated to reflux and stirred for 12 hours. The reaction solution was cooled to room temperature and adjusted with 1 M hydrochloric acid. The temperature was raised to pH <2, refluxed for 1 hour, cooled to room temperature, the reaction solution was concentrated under reduced pressure, the residue was poured into 200ml of water, adjusted to pH> 9 with 1M sodium hydroxide solution, and ethyl acetate (200mL × 2) Extraction, combined organic phases, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated the filtrate under reduced pressure to give the title compound 11c (2 g, yield: 78.81%).

    third step     N -((4- (4-bromophenyl) tetrahydro-2 H -pyran-4-yl) methyl) -2,2,2-trifluoroacetamidamine 11d

Dissolve 11c (2g, 7.40mmol) in 100ml of dichloromethane, lower the temperature to 0 ° C, add triethylamine (2.25g, 22.21mmol), slowly add trifluoroacetic anhydride (2.02g, 9.02mmol) dropwise, and raise to The reaction was stirred at room temperature for 12 hours. The reaction solution was poured into 100 mL of water, and the layers were separated. The organic phase was washed with saturated sodium bicarbonate and sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. It was purified by silica gel column chromatography with eluent system B The obtained residue gave the title compound 11d (2 g, yield: 73.78%).

    the fourth step     1- (7-bromo-2 ', 3', 5 ', 6'-tetrahydro-1 H -spiro [isoquinoline-4,4'-pyran] -2 (3 H ) -yl) -2 , 2,2-trifluoroethanone 11e

11d (2g, 5.46mmol) was dissolved in 50mL of a mixed solvent of preformed acetic acid and sulfuric acid (V / V = 2: 3), paraformaldehyde (492mg, 16.39mmol) was added, and the reaction was stirred for 12 hours. The reaction solution was poured into 300 mL of ice water and extracted with ethyl acetate (100 mL × 2). The organic phases were combined, washed with water, saturated sodium bicarbonate solution and saturated sodium chloride solution in that order, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography with eluent system B to obtain the title compound 11e (600 mg, yield: 29.05%).

    the fifth step     2- (2,2,2-trifluoroethenyl) -2,2 ', 3,3', 5 ', 6'-hexahydro-1 H -spiro [isoquinoline-4,4'-pyridine (Nan) -7-carboxylic acid ethyl ester 11f

Compound 11e (600 mg, 1.59 mmol) was dissolved in 20 mL of a mixed solvent of dimethylarsine (V / V = 1: 1), and 1,3-bis (diphenylphosphine) propane (130 mg, 317.31 μmol) was added. ), Triethylamine (481 mg, 4.76 mmol) and palladium acetate (71 mg, 317.71 μmol), under a carbon monoxide atmosphere, the temperature was raised to 60 ° C. and the reaction was stirred for 12 hours. The reaction solution was cooled to room temperature, poured into 100 mL of water, and extracted with ethyl acetate (100 mL × 2). The organic phases were combined, washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by column chromatography with eluent system B to obtain the title compound 11f (100 mg, yield: 16.97%).

    Step Six     11g of 2,2 ', 3,3', 5 ', 6'-hexahydro-1 H -spiro [isoquinoline-4,4'-pyran] -7-carboxylic acid ethyl ester

11f (100 mg, 269.29 μmol) was dissolved in 50 mL of a mixed solvent of ethanol and water (V / V = 1: 1), potassium carbonate (111 mg, 807.86 μmol) was added, and the reaction was stirred at room temperature for 12 hours. The reaction solution was poured into 30 mL of water and extracted with ethyl acetate (50 mL × 2). The organic phases were combined, washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain 11 g (70 mg) of the crude title compound. The product was directly subjected to the next reaction without purification.

    Step Seven     2- (4- (trifluoromethyl) phenyl) -2,2 ', 3,3', 5 ', 6'-hexahydro-1 H -spiro [isoquinoline-4,4'-pyran ] -7-carboxylic acid ethyl ester 11h

11 g (50 mg, 181.59 μmol) of crude compound and 1-bromo-4- (trifluoromethyl) benzene 1e (61 mg, 272 μmol) were prepared by a well-known method " Organic Letters , 2014, 16 (16), 4268-4271" (Obtained) was dissolved in 30 mL of 1,4-dioxane, and cesium carbonate (177 mg, 544.78 μmol) and 2-dicyclohexyl phosphorus-2,4,6-triisopropylbiphenyl (8 mg, 18.16 μmol) were added. And tris (dibenzylideneacetone) dipalladium (16 mg, 18.16 μmol), and the reaction was stirred for 3 hours while heating to 90 ° C. under the protection of helium. The reaction solution was cooled to room temperature, poured into 50 mL of water, and extracted with ethyl acetate (50 mL × 2). The organic phases were combined, washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by column chromatography with eluent System B to obtain the title compound 11h (30 mg, yield: 39.39%).

    Step eight     2- (4- (trifluoromethyl) phenyl) -2,2 ', 3,3', 5 ', 6'-hexahydro-1 H -spiro [isoquinoline-4,4'-pyran ] -7-carboxylic acid 11i

Compound 11h (30 mg, 71.52 μmol) was dissolved in 30 mL of a mixed solvent of ethanol and water (V / V = 5: 1), sodium hydroxide (14 mg, 357.62 μmol) was added, and the mixture was heated to reflux and stirred for 2 hours. The reaction solution was cooled to room temperature, and concentrated under reduced pressure. The resulting residue was added dropwise 1 M hydrochloric acid to pH <2, and extracted with ethyl acetate (50 mL × 2). The organic phases were combined, washed with saturated sodium chloride solution, and anhydrous sulfuric acid. Sodium was dried, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude title compound 11i (30 mg). The product was directly subjected to the next reaction without purification.

    Step Nine     N- (4- (ethylsulfonyl) benzyl) -2- (4- (trifluoromethyl) phenyl) -2,2 ', 3,3', 5 ', 6'-hexahydro- 1 H -spiro [isoquinoline-4,4'-pyran] -7-formamidine 11

The crude compound 11i (30 mg, 76.65 μmol) and (4- (ethylsulfonyl) phenyl) methylamine for 1 h (18 mg, 91.98 μmol, prepared by the method disclosed in the patent application “WO2015017335”) were dissolved in 10 mL of N , To N -dimethylformamide, 2- (7-azabenzotriazole) -N, N, N ', N'-tetramethylurea hexafluorophosphate (43 mg, 114.98 μmol) and three Ethylamine (23 mg, 229.95 μmol) was stirred for 12 hours. The reaction solution was poured into 50 mL of water and extracted with ethyl acetate (50 mL × 2). The organic phases were combined, washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained product was purified by high performance liquid chromatography. The residue gave the title compound 11 (15 mg, yield: 34.17%).

MS m / z (ESI): 573.6 [M + 1]

¹ H NMR (400MHz, DMSO- d ₆ ) δ 9.11 (s, 1H), 7.78-7.80 (m, 4H), 7.53-7.62 (m, 5H), 7.15-7.16 (m, 2H), 4.53 (s, 4H), 3.73-7.75 (m, 6H), 2.04 (m, 2H), 1.49-1.52 (m, 2H), 1.20 (m, 2H), 1.05 (m, 3H).

    Example 12     N- (4- (ethylsulfonyl) benzyl) -2 '-(5- (trifluoromethyl) pyridin-2-yl) -2', 3'-dihydro-1 ' H -spiro [ Cyclopropane-1,4'-isoquinoline] -7'-formamidine 12

    First step     2 '-(5- (trifluoromethyl) pyridin-2-yl) -2', 3'-dihydro-1 ' H -spiro [cyclopropane-1,4'-isoquinoline] -7'- Ethyl carboxylate 12b

The crude compound 1d (500 mg, 2.16 mmol) and 2-chloro-5- (trifluoromethyl) pyridine 12a (732 mg, 3.24 mmol) were dissolved in 50 mL of 1,4-dioxane, and cesium carbonate (2.11 g, 6.49) was added. mmol), 2-dicyclohexylphosphine-2,4,6-triisopropylbiphenyl (103 mg, 216.82 μmol) and tris (dibenzylideneacetone) dipalladium (197 mg, 216.18 μmol), warmed to 90 ° C The reaction was stirred under helium for 3 hours. The reaction solution was cooled to room temperature, poured into 100 mL of water, and extracted with ethyl acetate (100 mL × 2). The organic phases were combined, washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by column chromatography with eluent system B to obtain the title compound 12b (450 mg, yield: 55.31%).

    Second step     2 '-(5- (trifluoromethyl) pyridin-2-yl) -2', 3'-dihydro-1 ' H -spiro [cyclopropane-1,4'-isoquinoline] -7'- Carboxylic acid 12c

Compound 12b (1.5 g, 3.99 mmol) was dissolved in 60 mL of a mixed solvent of ethanol and water (V / V = 5: 1), sodium hydroxide (797.12 mg, 19.93 mmol) was added, and the mixture was heated to reflux and stirred for 2 hours. The reaction solution was cooled to room temperature, and concentrated under reduced pressure. The resulting residue was added dropwise with 1 M hydrochloric acid to pH <2, and extracted with ethyl acetate (50 mL × 2). The organic phases were combined, washed with a saturated sodium chloride solution, and anhydrous sulfuric acid. Sodium was dried, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude title compound 12c (1.3 g). The product was directly subjected to the next reaction without purification.

    third step     N - (4- (ethyl-sulfo acyl) benzyl) -2 '- (5- (trifluoromethyl) pyridin-2-yl) -2', 3'-dihydro -1 'H - spiro [ Cyclopropane-1,4'-isoquinoline] -7'-formamidine 12

The crude compound 12c (1 g, 2.87 mmol) and (4- (ethylsulfonyl) phenyl) methylamine for 1 h (858 mg, 4.31 mmol, prepared using the method disclosed in the patent application "WO2015017335"-obtained) were dissolved in 30 mL of N , N -dimethylformamide, 2- (7-azabenzotriazole) -N, N, N ', N'-tetramethylurea hexafluorophosphate (1.64g, 4.31mmol) And triethylamine (871 mg, 8.61 mmol) and stirred for 12 hours. The reaction solution was poured into 100 mL of water and extracted with ethyl acetate (100 mL × 2). The organic phases were combined, washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained product was purified by high performance liquid chromatography. The residue was obtained as the title compound 12 (1 g, yield: 65.77%).

MS m / z (ESI): 530.4 [M + 1]

¹ H NMR (400MHz, DMSO- d ₆ ) δ 9.12-9.15 (m, 1H), 8.43 (s, 1H), 7.81-7.86 (m, 4H), 7.71-7.73 (m, 1H), 7.57-7.59 ( m, 2H), 7.07-7.09 (m, 1H), 6.97-6.99 (m, 1H), 4.99 (s, 2H), 4.58-4.59 (m, 2H), 3.83 (s, 2H), 3.24-3.30 ( m, 2H), 1.07-1.11 (m, 7H).

    Example 13     (R) - N - (1- (5- ( ethyl-sulfo acyl) pyridin-2-yl) -2-hydroxyethyl) -2 '- (4- (trifluoromethyl) phenyl) -2 ', 3'-Dihydro-1' H -spiro [cyclopropane-1,4'-isoquinoline] -7'-formamidine 13

1 g (50 mg, 143.96 μmol) of crude compound and ( R ) -2-amino-2- (5- (ethylsulfonyl) pyridin-2-yl) ethanol 13a (39 mg, 172.75 μmol, using the patent application " WO2016061160 "(prepared by the method disclosed) is dissolved in 10mL N , N -dimethylformamide, and 2- (7-azabenzotriazole) -N, N, N ', N'-tetramethyl is added Hexafluorophosphate (82 mg, 215.93 μmol) and triethylamine (43 mg, 431.87 μmol) were stirred for 12 hours. The reaction solution was poured into 30 mL of water and extracted with ethyl acetate (30 mL × 2). The organic phases were combined, washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained product was purified by high performance liquid chromatography. The residue was obtained as the title compound 13 (20 mg, yield: 24.83%).

MS m / z (ESI): 560.6 [M + 1]

¹ H NMR (400MHz, DMSO- d ₆ ) δ 8.98 (s, 1H), 8.43 (s, 1H), 8.73-8.75 (m, 1H), 8.23-8.24 (m, 1H), 7.83 (s, 1H) , 7.67-7.72 (m, 2H), 7.50-7.51 (m, 2H), 7.13-7.14 (m, 1H), 6.94-6.96 (m, 1H), 5.21 (s, 1H), 4.67 (s, 2H) , 3.85-3.87 (m, 3H), 3.55 (s, 2H), 3.39-3.40 (m, 2H), 1.05-1.13 (m, 7H).

    Example 14     N- (4- (ethylsulfonyl) benzyl) -6 '-(4- (trifluoromethyl) phenyl) -6', 7'-dihydro-5 ' H -spiro [cyclopropane- 1,8 '-[1,6] naphthyridine] -3'-formamidine 14

    First step     3'-nitro-5 ' H -spiro [cyclopropane-1,8'-[1,6] naphthyridine] -6 '(7' H ) -carboxylic acid third butyl ester 14b

Compound 14a (600 mg, 2.66 mmol prepared by the method disclosed in the patent application "WO2010114957") was dissolved in 30 ml of a 7M amine methanol solution, and compound 14b (530 mg, 2.66 mmol) was added to raise the temperature to 60 ° C, and the reaction was stirred overnight. The reaction solution was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography using eluent system B to obtain the title compound 14c (300 mg, yield: 36.89%).

    Second step     3'-amino -5 'H - spiro [cyclopropane-1,8' - [1,6] naphthyridine] -6 '(7' H) - acid tert-butyl ester 14d

Compound 14c (300mg, 982.54mol) was dissolved in a mixed solvent of 50ml ethanol and water (V / V = 2: 1), iron powder (274mg, 4.90mmol), ammonium chloride (106mg, 1.96mmol) were added, and the temperature was raised. The reaction was stirred at reflux for 3 hours. The reaction solution was cooled to room temperature. The reaction solution was poured into 50 ml of water and extracted with dichloromethane (100 mL × 2). The organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to give the crude title compound 14d (270 mg).

    third step     3'-bromo-5 ' H -spiro [cyclopropane-1,8'-[1,6] naphthyridine] -6 '(7' H ) -carboxylic acid third butyl ester 14e

Dissolve 14d (250mg, 907.95μmol) in 30ml acetonitrile, lower the temperature to 0 ° C, slowly add isoamyl nitrite (212mg, 1.81mmol) dropwise, stir for 30 minutes after adding, add cuprous bromide, and warm to room temperature The reaction was stirred for 12 hours. The reaction solution was poured into 50 mL of water and extracted with ethyl acetate (50 mL × 2). The organic phases were combined, washed with water, saturated sodium bicarbonate solution and saturated sodium chloride solution in that order, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography with eluent System B to obtain the title compound 14e (100 mg, yield: 32.46%).

    the fourth step     5 ' H -spiro [cyclopropane-1,8'-[1,6] naphthyridine] -3 ', 6' (7 ' H ) -dicarboxylic acid 6'-(third butyl) ester 3'- Ethyl ester 14f

Compound 14e (100 mg, 294.78 μmol) was dissolved in a mixed solvent of 20 mL of ethanol and dimethylarsine (V / V = 1: 1), and 1,3-bis (diphenylphosphine) propane (25 mg, 57.25 μmol) was added. ), Potassium carbonate (61 mg, 442.03 μmol) and palladium acetate (6 mg, 26.72 μmol), and the reaction was stirred at 60 ° C. for 12 hours under a carbon monoxide atmosphere. The reaction solution was cooled to room temperature, poured into 30 mL of water, and extracted with ethyl acetate (30 mL × 2). The organic phases were combined, washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by column chromatography with eluent System B to obtain the title compound 14f (80 mg, yield: 81.64%).

    the fifth step     6 ', 7'-dihydro -5' H - spiro [cyclopropane-1,8 '- [1,6] naphthyridine] -3'-carboxylate 14g

14f (80 mg, 240.68 μmol) was dissolved in a mixed solvent of 20 mL of ethanol and water (V / V = 1: 1), 5 ml of a 4M hydrochloric acid methanol solution was added, and the reaction was stirred at room temperature for 12 hours. The reaction solution was concentrated under reduced pressure, the residue was adjusted to pH> 8 with 1N NaOH solution, and extracted with ethyl acetate (30mL × 2). The organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate. Concentrated under reduced pressure to give 14 g (70 mg) of the crude title compound. The product was subjected to the next reaction without purification.

    Step Six     6 '- (4- (trifluoromethyl) phenyl) -6', 7'-dihydro -5 'H - spiro [cyclopropane-1,8' - [1,6] naphthyridine] -3 ' -Ethyl carboxylate 14h

14 g (70 mg, 301.36 μmol) of the crude compound and 1-bromo-4- (trifluoromethyl) benzene 1e (101 mg, 448.87 μmol) were prepared by a known method “ Organic Letters , 2014, 16 (16), 4268-4271” (Prepared) was dissolved in 20 mL of 1,4-dioxane, and cesium carbonate (294 mg, 902.34 μmol) was added, and 2-dicyclohexyl phosphorus-2,4,6-triisopropylbiphenyl (14 mg, 29.37 μmol) was added. ) And tris (dibenzylideneacetone) dipalladium (27 mg, 29.48 μmol), the reaction was stirred for 3 hours while heating to 90 ° C. under the protection of helium. The reaction solution was cooled to room temperature, poured into 30 mL of water, and extracted with ethyl acetate (30 mL × 2). The organic phases were combined, washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by column chromatography with eluent System B to give the title compound 14h (100 mg, yield: 88.16%).

    Step Seven     6 '- (4- (trifluoromethyl) phenyl) -6', 7'-dihydro -5 'H - spiro [cyclopropane-1,8' - [1,6] naphthyridine] -3 ' -Carboxylic acid 14i

Compound 14h (100 mg, 265.69 μmol) was dissolved in 20 mL of a mixed solvent of ethanol and water (V / V = 5: 1), sodium hydroxide (53 mg, 1.32 mmol) was added, and the mixture was heated to reflux and stirred for 2 hours. The reaction solution was cooled to room temperature, and concentrated under reduced pressure. The resulting residue was added dropwise with 1 M hydrochloric acid to pH <2, and extracted with ethyl acetate (50 mL × 2). The organic phases were combined, washed with a saturated sodium chloride solution, and anhydrous sulfuric acid. Sodium was dried, filtered, and the filtrate was concentrated under reduced pressure to give the crude title compound 14i (80 mg). The product was directly subjected to the next reaction without purification.

    Step eight     N- (4- (ethylsulfonyl) benzyl) -6 '-(4- (trifluoromethyl) phenyl) -6', 7'-dihydro-5 ' H -spiro [cyclopropane- 1,8 '-[1,6] naphthyridine] -3'-formamidine 14

The crude compound 14i (80 mg, 229.67 μmol) and (4- (ethylsulfonyl) phenyl) methylamine for 1 h (68 mg, 341.24 μmol, prepared by the method disclosed in the patent application “WO2015017335”) were dissolved in 10 mL of N , To N -dimethylformamide, 2- (7-azabenzotriazole) -N, N, N ', N; -tetramethylurea hexafluorophosphate (130 mg, 342.10 μmol) and three Ethylamine (69 mg, 681.88 μmol) was stirred for 12 hours. The reaction solution was poured into 30 mL of water and extracted with ethyl acetate (30 mL × 2). The organic phases were combined, washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained product was purified by high performance liquid chromatography. The residue gave the title compound 14 (30 mg, yield: 24.66%).

MS m / z (ESI): 530.2 [M + 1]

¹ H NMR (400MHz, DMSO- d ₆ ) δ 9.27-9.29 (m, 1H), 8.81-8.82 (m, 1H), 8.09-8.10 (m, 1H), 7.84-7.86 (m, 2H), 7.58- 7.60 (m, 2H), 7.51-7.53 (m, 2H), 7.18-7.21 (m, 2H), 4.73 (s, 2H) 4.60-4.61 (m, 2H), 3.74 (s, 2H), 3.25-3.30 (m, 2H), 1.26-1.28 (m, 2H), 1.07-1.09 (m, 5H).

Biological evaluation

The present invention is further described below in conjunction with test examples, but these examples are not meant to limit the scope of the present invention.

Test Example 1. Determination of RORγ in vitro activity of the compound of the present invention

I. Experimental materials and instruments

1. LanthaScreen® TR-FRET RORγ Co-Startup System (Life Technologies)

2. RORγ LBD (AB Vector)

3. DMSO (SigmaAldrich)

4. Microplate reader (Tecan)

Experimental steps

The LanthaScreen TR-FRET (Time-Resolved Fluorescence Energy Resonance Transfer) RORγ co-start system was used to screen the compounds of the present invention for modulation of RORγ activity.

First complete TR-FRET Coregulator (Life Technologies) was prepared to contain a final concentration of 5 mM DTT. The final DMSO concentration was 2%. The test compound was serially diluted to a 2x final concentration in complete buffer D containing 2% DMSO, and the highest dose was 60 μm. 10 μl / well was added to a test well (PerkinElmer) of a 384-well plate. Two parallel control wells were set up for each test compound at the same concentration. Prepare 4X RORγ LBD (AB Vector). RORγ LBD was diluted to 1 ng / μL using intact buffer D. 5 μl / well was added to the test wells of a 384-well assay plate. The negative control well was 5 μL of intact buffer D without RORγ LBD. A complete buffer D was used to prepare a mixed solution containing 0.6 μM luciferin-D22 (4X) and 8 nM 鋱 (Tb) labeled anti-GST antibody (4X) (Life Technologies), and 5 μL of the mixed solution was added to a 384-well plate. The total reaction system was 20 μL. Gently mix the 384-well plate on a shaker and incubate at room temperature in the dark for 2-4 hours.

Tecan Infinite M1000 was used to detect the fluorescence readings, and the graph of the emission wavelength ratio of 520nm / 495nm and the logarithm of the compound concentration was plotted by GraphPad Prism 6.0 software to calculate the IC ₅₀ / EC ₅₀ value of the test compound.

RORγ compounds of the invention in vitro activity was measured by the above tests, the measured IC ₅₀ / EC ₅₀ values are shown in Table 1.

a: If an inhibitor is, the value indicated as IC _50; if it is an agonist, the value denoted EC _50.

Conclusion: The compounds of the present invention have significant regulatory effects on the in vitro activity of RORγ. The experimental results show that the change of the A ring substituent in the compound represented by the general formula (I) shows a different mechanism for the regulation of RORγ in vitro activity. A has a smaller substituent (for example, a hydrogen atom) at the ortho position of A, and exhibits an inhibitory effect (see Examples 1 to 7 and 9 to 14 etc.), and has a larger substitution at the ortho position of ring A. Group (for example: trifluoromethyl), it shows an agonistic effect (see Example 8).

Test example 2. Determination of the activity of the compound of the present invention on IL-17A enzyme-linked immunoassay

I. Experimental materials and instruments

1. Human peripheral blood mononuclear cells (PBMC) (Zenbio)

2. Lymphocyte Medium (Zenbio)

3. TexMACS (Miltenyi Biotec)

4. Human Cytostim (Miltenyi Biotec)

5. Human IL-17 ELISA Kit (R & D System)

6. CO ₂ incubator (Fisher Scientific)

7. Centrifuge (Fisher Scientific)

8. 96-well cell culture plate (Fisher Scientific)

9. Microplate reader (Tecan)

Experimental steps

Frozen human peripheral blood mononuclear cells (PBMC) were quickly resuscitated in pre-warmed lymphocyte culture medium, centrifuged at 1000 rpm for 10 min, the cell culture supernatant was removed, and the cells were gently suspended in TexMACS medium to count the cells. T cell initiation reagent cytostim (10 μl / ml) was added to the cell suspension in proportion, and then cells were seeded in a 96-well cell culture plate at a density of 1 × 10 ⁵ peripheral blood mononuclear cells / well. TexMACS medium was used to gradiently dilute the test compounds and added to each experimental well, with 2-3 parallel wells in each group. Prepare negative control wells containing cells without cytostim to obtain background readings. The cell culture plate was placed in a 5% carbon dioxide 37 ° C incubator for 3 days. The cell culture supernatant was collected 3 days after the drug treatment, and the suspension was removed by centrifugation. IL-17A in the supernatant was then quantified using the IL-17A ELISA kit. Calculated using GraphPad Prism 6.0 ₅₀ / EC ₅₀ value of the test compound IC.

The IL-17A enzyme-linked immunological quantitative analysis of the compound of the present invention was determined by the above test. The IC ₅₀ values measured are shown in Table 2.

Conclusion: The compound of the present invention has obvious regulating effect on the activity of IL-17A enzyme-linked immunoassay.

Claims (26)

A compound represented by the general formula (AI) or a tautomer, a racemate, a racemate, an enantiomer, a diastereomer or a mixture thereof or a pharmaceutically acceptable salt thereof , Wherein: W ¹ , W ² and W ^{3 are the} same or different, and each is independently CH or N; ring A and ring B are the same or different, and each is independently selected from cycloalkyl, heterocyclyl, aryl, and hetero Aryl; ring C is a cycloalkyl or heterocyclic group; R ^{1 is the} same or different and each is independently selected from a hydrogen atom, halogen, alkyl, haloalkyl, alkoxy, haloalkoxy, cyano, Amine, nitro, hydroxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl, wherein the alkyl, haloalkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl Each independently selected from the group consisting of halogen, alkyl, haloalkyl, alkoxy, haloalkoxy, cyano, amino, nitro, hydroxyl, hydroxyalkyl, cycloalkyl, heterocyclyl, aromatic And heteroaryl are substituted with one or more substituents; R ^{2 is the} same or different, and each is independently selected from a hydrogen atom, halogen, alkyl, haloalkyl, alkoxy, haloalkoxy, cyano Group, amino group, nitro group, hydroxy group, hydroxyalkyl group, cycloalkyl group, heterocyclic group, aryl group, heteroaryl group, -OR ⁴ , -C (O) R ⁴ , -C (O) OR ⁴ and- S (O) _m R ⁴ ; R ^{3 is} selected from hydrogen atom, halogen, alkyl, halogen Alkyl, alkoxy, haloalkoxy, cyano, amino, nitro, hydroxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl, wherein the alkyl, haloalkyl , Cycloalkyl, heterocyclyl, aryl and heteroaryl are each independently selected from the group consisting of hydroxy, halogen, alkyl, haloalkyl, cyano, amine, nitro, alkoxy, haloalkoxy Substituted with one or more substituents of aryl, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl; R ^{4 is} selected from hydrogen atom, alkyl, cycloalkyl, haloalkyl, and alkane Oxy, haloalkoxy, heterocyclyl, aryl, and heteroaryl, wherein the alkyl, cycloalkyl, haloalkyl, heterocyclyl, aryl, and heteroaryl are each independently selected as necessary One of hydroxy, halogen, alkyl, haloalkyl, cyano, amine, nitro, alkoxy, haloalkoxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl Substituted with one or more substituents; m is 0, 1, or 2; s is 0, 1, 2, 3, or 4; and t is 0, 1, 2, 3, or 4. The compound represented by the general formula (AI) or its tautomer, meso, racemate, enantiomer, diastereomer or its In the form of a mixture or a pharmaceutically acceptable salt thereof, wherein ring C is a cycloalkyl group of C _3-6 or a heterocyclic group of 3 to 6 members, wherein the heterocyclic group contains 1 to 3 members selected from N, O or S Heteroatom.     The compound represented by the general formula (AI) or the tautomer, meso, racemate, enantiomer, diastereomer, or A mixture or a pharmaceutically acceptable salt thereof, wherein ring C is selected from cyclopropyl, cyclobutyl, cyclopentyl or tetrahydropyranyl.     The compound represented by the general formula (AI) or its tautomer, meso, racemate, enantiomer, diastereomer or its A mixture or a pharmaceutically acceptable salt thereof, which is a compound represented by the general formula (I) or a tautomer, a racemate, a racemate, an enantiomer, a diastereomer Or a mixture thereof or a pharmaceutically acceptable salt thereof: Wherein: n is 1, 2 or 3; ring A, ring B, R ¹ to R ³ , s and t are as defined in the first item of the patent application scope. The compound represented by the general formula (I) or the tautomer, meso, racemate, enantiomer, diastereomer, or A mixture or a pharmaceutically acceptable salt thereof, which is a compound represented by the general formula (II) or a tautomer, meso, racemate, enantiomer, diastereomer Or a mixture thereof or a pharmaceutically acceptable salt thereof: Among them: ring A, ring B, R ¹ to R ³ , s and t are as defined in item 4 of the scope of patent application.     The compound or a tautomer, meso, racemate, enantiomer, diastereomer or a mixture thereof according to any one of claims 1 to 5 of the scope of patent application Form, or a pharmaceutically acceptable salt thereof, wherein ring A is selected from phenyl, pyridyl, pyrimidinyl, cyclohexyl, and piperidinyl.     The compound represented by general formula (I) or its tautomer, meso, racemate, enantiomer, diastereomer Or a mixture thereof or a pharmaceutically acceptable salt thereof, which is a compound represented by the general formula (III) or a tautomer, meso, racemate, enantiomer, diastereomer Structure or mixture thereof or a pharmaceutically acceptable salt thereof: Wherein: G ¹ and G ^{2 are the} same or different, and are each independently CH or N; rings B, R ¹ to R ³ and t are as defined in item 4 of the scope of patent application. The compound represented by general formula (I) or its tautomer, meso, racemate, enantiomer, diastereomer Or a mixture thereof or a pharmaceutically acceptable salt thereof, which is a compound represented by the general formula (IV) or a tautomer, meso, racemate, enantiomer, diastereomer Structure or mixture thereof or a pharmaceutically acceptable salt thereof: Wherein: R ^{b is} selected from the group consisting of alkyl, haloalkyl, alkoxy, haloalkoxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl, wherein the alkyl, haloalkyl, Cycloalkyl, heterocyclyl, aryl, and heteroaryl are each independently selected from halogen, hydroxy, alkyl, haloalkyl, cyano, amine, nitro, alkoxy, haloalkoxy , Hydroxyalkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl with one or more substituents; p is 0, 1, 2 or 3; ring B, R ¹ ~ R ³ and t As defined in item 4 of the scope of patent application.     The compound or a tautomer, a racemate, a racemate, an enantiomer, a diastereomer, or a mixture thereof according to any one of claims 1 to 8 of the scope of patent application Or a pharmaceutically acceptable salt thereof, wherein ring B is selected from phenyl, pyridyl, pyrimidinyl, cyclohexyl, and piperidinyl.     The compound or a tautomer, a racemate, a racemate, an enantiomer, a diastereomer, or a mixture thereof according to any one of claims 1 to 9 of the scope of patent application Form, or a pharmaceutically acceptable salt thereof, wherein R ² is -S (O) _m R ⁴ ; m is 2; and R ⁴ is alkyl. The compound represented by the general formula (III) or the tautomer, meso, racemate, enantiomer, diastereomer, or mixture thereof described in item 7 of the scope of the patent application Form or a pharmaceutically acceptable salt thereof, which is a compound represented by the general formula (III-A) or a tautomer, meso, racemate, enantiomer, diastereomer Or in the form of a mixture or a pharmaceutically acceptable salt thereof: Wherein: R ⁴ is an alkyl group; R ¹ and R ³ are as defined in item 7 of the scope of patent application. The compound represented by the general formula (IV) or its tautomer, meso, racemate, enantiomer, diastereomer, or a mixture thereof as described in claim 8 Form or a pharmaceutically acceptable salt thereof, which is a compound represented by the general formula (IV-A) or a tautomer, meso, racemate, enantiomer, diastereomer Or in the form of a mixture or a pharmaceutically acceptable salt thereof: Where: R ^{b is} selected from the group consisting of alkyl, haloalkyl, alkoxy, haloalkoxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl, wherein the alkyl, haloalkyl, cyclic Alkyl, heterocyclyl, aryl, and heteroaryl are each independently selected from halogen, hydroxy, alkyl, haloalkyl, cyano, amine, nitro, alkoxy, haloalkoxy, Substituted with one or more substituents of hydroxyalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl; R ⁴ is alkyl; p is 0, 1, 2 or 3; R ¹ and R ³ As defined in item 1 of the patent application scope. The compound represented by the general formula (I) or any tautomer, meso, racemate, enantiomer, or Enantiomers or mixtures thereof, or a pharmaceutically acceptable salt thereof, wherein ^Rb is haloalkyl. The compound represented by the general formula (AI) as described in any one of claims 1 to 13 or a tautomer, meso, racemate, enantiomer, diastereomer enantiomers or mixtures or a pharmaceutically acceptable salt thereof, wherein R ¹ is haloalkyl. The compound represented by the general formula (AI) as described in any one of claims 1 to 14 or a tautomer, meso, racemate, enantiomer, diastereomer An enantiomer or a mixture thereof or a pharmaceutically acceptable salt thereof, wherein R ³ is a hydrogen atom, an alkyl group or a hydroxyalkyl group. The compound or the tautomer, meso, racemate, enantiomer, diastereomer, or mixture thereof as described in any one of claims 1 to 15 of the scope of patent application , Or a pharmaceutically acceptable salt thereof, selected from: A compound represented by the general formula (AI-A) or a tautomer, a racemate, a racemate, an enantiomer, a diastereomer or a mixture thereof or a pharmaceutically acceptable compound thereof salt, Among them: ring A, ring C, W ¹ , W ² , W ³ , R ¹ and s are as defined in the first scope of the patent application. The compound represented by general formula (AI-A) or its tautomer, meso, racemate, enantiomer, diastereomer, or A mixture thereof or a pharmaceutically acceptable salt thereof, which is a compound represented by the general formula (IA) or a tautomer, a racemate, a racemate, an enantiomer, a diastereomer Or a mixture thereof or a pharmaceutically acceptable salt thereof: Or a tautomer, a racemate, a racemate, an enantiomer, a diastereomer or a mixture thereof, or a pharmaceutically acceptable salt thereof, wherein: ring A, R ¹ , s And n are as defined in item 17 of the scope of patent application. The compound represented by the general formula (AI-A) or its tautomer, meso, racemate, enantiomer, diastereomer A conformation or a mixture thereof or a pharmaceutically acceptable salt thereof, which is selected from A method for preparing a compound of general formula (AI) or a tautomer, a racemate, a racemate, an enantiomer, a diastereomer or a mixture thereof as described in item 1 of the scope of patent application A method of form or a pharmaceutically acceptable salt thereof, the method comprising: A compound of the general formula (AI-A) and a compound of the general formula (IB) undergo a condensation reaction to obtain a compound of the general formula (AI); wherein: ring A, ring B, ring C, W ¹ , W ² , W ³ , R ¹ ~ R ³ , s and t are as defined in item 1 of the scope of patent application. A method for preparing a compound of the general formula (I) or a tautomer, a racemate, a racemate, an enantiomer, a diastereomer or a mixture thereof as described in item 4 of the scope of patent application A method of form or a pharmaceutically acceptable salt thereof, the method comprising: A compound of the general formula (IA) and a compound of the general formula (IB) undergo a condensation reaction to obtain a compound of the general formula (I); wherein: ring A, ring B, R ¹ to R ³ , n, s, and t are as in the first patent application As defined in the item.     A medicinal composition containing a therapeutically effective amount of a compound represented by the general formula (I) according to any one of claims 1 to 16 or a tautomer, meso, racemic Rotates, enantiomers, diastereomers, or mixtures thereof, or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers, diluents, or excipients.         A compound represented by general formula (I) or a tautomer, meso, racemate, enantiomer, or The use of an enantiomer or a mixture thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition described in item 22 of the scope of patent application, which is used to prepare a ROR modulator.         A compound represented by general formula (I) or a tautomer, meso, racemate, enantiomer, or Use of enantiomers or mixtures thereof or pharmaceutically acceptable salts thereof or the pharmaceutical composition described in item 22 of the scope of patent application, which are used for the preparation and prevention of and / or treatment of inflammation, autoimmune diseases, tumors Or cancer drugs.         A compound represented by general formula (I) or a tautomer, meso, racemate, enantiomer, or Enantiomers or mixtures thereof, or pharmaceutically acceptable salts thereof, or the use of a medicinal composition described in item 22 of the scope of patent application, which are used as ROR inhibitors in the preparation for the prevention and / or treatment of inflammation or autoimmunity Drug for sexually transmitted diseases, the inflammation or autoimmune disease is selected from asthma, atopic dermatitis, contact dermatitis, acne, bronchitis, Crohn's disease, localized enteritis, inflammatory bowel disease (IBD), ulcerative Colitis, allograft rejection, Hugh's syndrome, uveitis, Behcet's disease, dermatomyositis, multiple sclerosis, ankylosing spondylitis, neuromyelitis, systemic lupus erythematosus (SLE), scleroderma, pancreatitis, psoriasis, psoriatic arthritis (PsA), rheumatoid arthritis, arthritis, osteoarthritis, allergic rhinitis, autoimmune diabetes, and autoimmune thyroid disease .         A compound represented by general formula (I) or a tautomer, meso, racemate, enantiomer, or Enantiomers or mixtures thereof, or pharmaceutically acceptable salts thereof, or the use of a pharmaceutical composition described in item 22 of the scope of patent application, which are used as ROR agonists in the preparation of tumors or cancers for prevention and / or treatment Drug, the tumor or cancer is selected from non-Hodgkin's lymphoma, diffuse large B-cell lymphoma, follicular lymphoma, synovial sarcoma, breast cancer, cervical cancer, colon cancer, lung cancer, gastric cancer, rectal cancer, pancreatic cancer , Brain cancer, skin cancer, oral cancer, prostate cancer, bone cancer, kidney cancer, ovarian cancer, bladder cancer, liver cancer, fallopian tube tumor, ovarian tumor, peritoneal tumor, stage IV melanoma, solid tumor, glioma, neuroglia Blastoma, hepatocellular carcinoma, mastoid nephroma, head and neck tumor, leukemia, lymphoma, myeloma, and non-small cell lung cancer.