Application of pyrazolopyridine compound in preparation of medicine for treating BTK related diseases including central nervous system diseases

Abstract

The invention discloses application of # imgabs0 # compounds shown in a formula (I) in preparation of drugs for treating BTK related diseases including central nervous system diseases, and belongs to the technical field of chemical medicines. The compound as shown in the formula (I) has high activity and high selectivity on BTK kinase, has a remarkable treatment effect on BTK related diseases such as lymphoma, leukemia, autoimmune diseases, inflammatory diseases or metabolic diseases, and has a synergistic interaction effect with various types of compounds and medicines with different mechanisms; particularly, the compound can effectively penetrate through the blood brain barrier and keep high concentration in the brain tissue, so that the treatment effect on the diseases of the central nervous system can be achieved; through cell experiments, the compound shown in the formula (I) shows a remarkable effect in treating central nervous system diseases such as multiple sclerosis and central nervous system lymphoma, and can become a novel method for treating the central nervous system diseases.

Description

Use of pyrazolopyridine compounds in the preparation of drugs for treating BTK-related diseases including central nervous system diseases

[0001] This application is based on the Chinese patent application with application number CN202210903508.7 and application date July 28, 2022, and claims the priority of the Chinese patent application. The entire content of the Chinese patent application is hereby introduced into this application as a reference. Technical Field

[0002] The present invention belongs to the field of chemical medicine technology, and specifically relates to the use of a pyrazolopyridine compound in preparing a drug for treating BTK-related diseases, and more particularly to the use of a pyrazolopyridine compound in preparing a drug for treating central nervous system diseases. Background Art

[0003] Bruton's tyrosine kinase (BTK) is a key kinase in the B cell receptor (BCR) signaling pathway. Irreversible BTK inhibitors covalently bind to the kinase's active site, Cys-481, inhibiting BTK activity, thereby effectively suppressing B cell hyperproliferation and achieving anti-tumor or anti-inflammatory effects. As a key kinase in the B cell receptor signaling pathway, BTK is crucial for the development and function of immune cells involved in autoimmune diseases and lymphoma pathogenesis, including B lymphocytes, macrophages, and microglia. Among currently marketed drugs, ibrutinib, an irreversible BTK inhibitor jointly developed by Pharmacyclis and Johnson & Johnson, is FDA-approved for the treatment of mantle cell lymphoma, chronic lymphocytic leukemia, Waldenstrom's macroglobulinemia, and chronic graft-versus-host disease. However, ibrutinib also has strong inhibitory effects on other kinases besides BTK. Inhibition of EGFR, ITK, and TEC kinases in particular can lead to severe adverse reactions such as rash, diarrhea, and bleeding. Therefore, there is an urgent need to develop a highly active BTK inhibitor with good kinase selectivity for the treatment of lymphoma and autoimmune diseases.

[0004] Furthermore, some diseases that primarily affect the brain are among the most prevalent, devastating, and poorly treated of all diseases. Despite tremendous advances in basic neuroscience knowledge, the failure rate of new drugs for important central nervous system (CNS) diseases continues to exceed that of most other areas of drug discovery. The blood-brain barrier (BBB) ​​is a significant obstacle to drug development for these diseases. Formed by the endothelium of brain capillaries, the BBB is a dynamic diffusion barrier that is crucial for protecting and maintaining brain homeostasis. The BBB excludes more than 100% of large molecule therapeutics and more than 98% of small molecule drugs from the brain, resulting in the lack of effective treatment options for the vast majority of CNS diseases. Furthermore, in addition to the permeability issue, even if a compound is able to cross the barrier, it must maintain therapeutically relevant concentrations at the target site to elicit its desired effect.

[0005] Therefore, there is a need in this field to develop a new class of BTK inhibitors that are highly active, can penetrate the blood-brain barrier, and possess good kinase selectivity for the treatment of central nervous system diseases, including lymphomas and autoimmune diseases. BTK inhibitors also exhibit synergistic effects with a variety of compounds and drugs with different mechanisms of action, and their combined use has demonstrated promising results for various of the aforementioned diseases.

[0006] Summary of the Invention

[0007] The compound of formula (I) of the present invention has high activity and selectivity against BTK kinase and has significant therapeutic effects on BTK-related diseases such as lymphoma, leukemia, autoimmune diseases, inflammatory diseases or metabolic diseases. In addition, central nervous system diseases have always been a difficult problem in the medical field due to their special characteristics. The present invention has found that pyrazolopyridine compounds can effectively penetrate the blood-brain barrier and maintain high concentrations in brain tissue. Therefore, the compound can achieve a therapeutic effect on diseases of the central nervous system.

[0008] The present invention provides the use of a compound represented by formula (I), an isomer thereof, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof (as known in the art, a pharmaceutical composition refers to a compound, an isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient and a pharmaceutically acceptable excipient in a therapeutically effective amount) in the preparation of a medicament for preventing or treating BTK-related diseases including central nervous system diseases, wherein the BTK-related diseases include lymphoma, leukemia, autoimmune diseases, inflammatory diseases, or metabolic diseases;

[0009] The structure of the compound of formula (I) is as follows:

[0010] in,

[0011] T1 is independently selected from N and CH;

[0012] R2 is independently selected from H, C 1-6 Alkyl, C 2-6 Alkenyl and C 2-6 Alkynyl, the C 1-6 Alkyl, C 2-6 Alkenyl and C 2-6 Alkynyl is optionally substituted by 1, 2 or 3 R a replace;

[0013] Ring A is selected from phenyl and 5-6 membered heteroaryl;

[0014] M is independently selected from C 3-6 Cycloalkyl and 3-6 membered heterocycloalkyl, the C 3-6 Cycloalkyl and 3-6 membered heterocycloalkyl are each independently optionally substituted by 1, 2 or 3 R b replace;

[0015] R1 and R3 are independently selected from F, Cl, Br, I, OH, NH2, CN;

[0016] n and m are independently selected from 0, 1, 2 or 3, and n and m are not 0 at the same time;

[0017] L1 and L2 are independently selected from -CH2-, -CH2CH2-, -O-, -C(=O)- and -C(=O)-NH-;

[0018] R a Independently selected from H, F, Cl, Br, I, OH, NH2, CN, C 1-3 Alkyl, C 1-3 Alkoxy and C 1-3 Alkylamino, the C 1-3 Alkyl, C 1-3 Alkoxy and C 1-3 Alkylamino is optionally substituted with 1, 2 or 3 R groups independently;

[0019] R b Selected from F, Cl, Br, I, CH3;

[0020] R is selected from H, F, Cl, Br, I;

[0021] The 5-6 membered heteroaryl and 3-6 membered heterocycloalkyl groups each independently contain 1, 2, 3 or 4 heteroatoms or heteroatom groups independently selected from -NH-, -O-, -S-, -C(=O)-, -S(=O)-, and N.

[0022] Among them, in the above-mentioned use, the BTK-related diseases include central nervous system diseases and non-central nervous system diseases.

[0023] Among them, in the above-mentioned uses, the central nervous system diseases include central nervous system lymphoma, central nervous system diffuse large B-cell lymphoma, Burkitt lymphoma arising in the central nervous system, autoimmune encephalitis, multiple sclerosis or neuromyelitis optica spectrum disease.

[0024] Among them, in the above-mentioned uses, the non-central nervous system diseases include B-cell lymphoma, chronic lymphocytic leukemia / small lymphocytic lymphoma, mantle cell lymphoma, Waldenstrom's macroglobulinemia, marginal zone lymphoma, follicular lymphoma, Burkitt lymphoma arising in the collar bone, craniofacial bones or abdominal organs, non-Hodgkin's lymphoma, diffuse large B-cell lymphoma, multiple myeloma, rheumatoid arthritis, systemic lupus erythematosus, lupus nephritis, Sjögren's syndrome, IgG4-related diseases, chronic spontaneous urticaria, idiopathic thrombocytopenic purpura, immune thrombocytopenia, pemphigus, asthma, graft-versus-host disease, scleroderma, new coronavirus infection, granulomatosis with polyangiitis, idiopathic pulmonary fibrosis, psoriasis, atherosclerosis or diabetes.

[0025] Burkitt lymphoma is a highly malignant B-cell tumor that likely arises from follicular germinal center cells. It occurs primarily in African children. It is more common in males than females, and in my country, it primarily affects children and young adults. The histological hallmark of this Epstein-Barr virus-associated tumor is a diffuse, medium-sized lymphoid cell composition with scattered macrophages phagocytizing debris between the tumor cells, forming a so-called starry sky pattern. It is most common in the collarbone, craniofacial bones, abdominal organs, and central nervous system, generally sparing peripheral lymph nodes and the spleen. Leukemic features are rare.

[0026] Among the above-mentioned non-central nervous system diseases, B-cell lymphoma includes non-Hodgkin's lymphoma, and non-Hodgkin's lymphoma includes diffuse large B-cell lymphoma, follicular lymphoma, marginal zone lymphoma, Waldenstrom's macroglobulinemia, mantle cell lymphoma, chronic lymphocytic leukemia / small lymphocytic lymphoma, and Burkitt's lymphoma.

[0027] On the basis of the above, the present invention also provides the use of the compound represented by formula (I), its isomer, its pharmaceutically acceptable salt or its pharmaceutical composition, in combination with at least one second drug for preparing a drug for preventing or treating BTK-related diseases including central nervous system diseases; the second drug is selected from CD-20 inhibitors or antibodies, BCL-2 inhibitors, SYK inhibitors, PI3K inhibitors, immunosuppressants, chemotherapy drugs or autoimmune checkpoint inhibitors.

[0028] The compound represented by formula (I) and the second drug can be administered continuously (the compound represented by formula (I) can be administered first, or the second drug can be administered first) or simultaneously. Therefore, when the compound represented by formula (I) is combined with the second drug, the compound represented by formula (I) and the second drug can be prepared as a whole or as separate entities.

[0029] Wherein, in the above use, the mass ratio of the compound represented by formula (I) to the second drug is 1-100:100-1.

[0030] Preferably, in the above use, the mass ratio of the compound represented by formula (I) to the second drug is 1-10:10-1.

[0031] Among them, in the above use, the CD-20 inhibitor or antibody includes rituximab, utoximab, ocrelizumab, ibritumomab tiuxetan, obinutuzumab, ofatumumab or vedolizumab.

[0032] Among them, in the above use, the BCL-2 inhibitor includes venetoclax, APG-2575, BGB-11417, ICP-248, FCN-338 or LP-108.

[0033] Among them, in the above use, the SYK inhibitor includes fostamatinib, entospletinib, PRT-2761, HMPL-523, SKI-O-703, IC-265 or GSK-2646264.

[0034] Among them, in the above-mentioned use, the PI3K inhibitors include copanlisib, erbulisib, durensibu, adranib, alpelisib, parsaclisib, limalisib, eganelisib, BAY 10-82439, buparlisib, dactolisib, bifentinib or puquintinib.

[0035] Among them, in the above-mentioned use, the immunosuppressant includes lenalidomide, pomalidomide, and thalidomide.

[0036] Among them, in the above-mentioned use, the chemotherapy drugs include temozolomide, cyclophosphamide, chlorambucil, bendamustine, ifosfamide, prednisone, dexamethasone, cisplatin, carboplatin, oxaliplatin, fludarabine, pentostatin, cladribine, cytarabine, gemcitabine, methotrexate, pralatrexate, anthracyclines, doxorubicin, liposomal doxorubicin, vincristine, mitoxantrone, etoposide or bleomycin.

[0037] Among them, in the above-mentioned uses, the autoimmune checkpoint inhibitors include pembrolizumab, nivolumab, carrelizumab, toripalimab, cemiplimab, durvalumab, velumab, sintilimab, tilelizumab, sepalimab, durvalumab, atezolizumab, envolimab, sugemalimab, sluglizumab, penampalimab, ceruletumab, zimbalimab, gepnolimab, badalizumab, proglitumab, pucometinib, cosibelimab, AUNP-12, reluzumab, retivarizumab, dotalimab, cadonimab, socamuzumab, adebrizumab, ipilimumab or tremelimumab.

[0038] Among them, in the above uses, R a Independently selected from H, F, Cl, Br, I, OH, NH2, CN, CH3, OCH3, NH(CH3)2 and N(CH3)2.

[0039] Wherein, in the above uses, R2 is independently selected from H, C 1-3 Alkyl, C 2-4 Alkenyl and C 2-4 Alkynyl, the C 1-3 Alkyl, C 2-4 Alkenyl and C 2-4 Alkynyl is optionally substituted by 1, 2 or 3 R a replace.

[0040] Preferably, in the above uses, R2 is independently selected from H, CH3, vinyl and propynyl.

[0041] Wherein, in the above use, M is independently selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, azetidinyl, oxetanyl, piperidinyl and morpholinyl, and the cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, azetidinyl, oxetanyl, piperidinyl and morpholinyl are independently optionally replaced by 1, 2 or 3 R b replace.

[0042] Preferably, in the above use, M is independently selected from piperidinyl and morpholinyl.

[0043] Wherein, in the above uses, L1 is selected from -O- and -C(=O)-NH-.

[0044] Wherein, in the above uses, L2 is selected from -C(=O)- and -C(=O)-NH-.

[0045] Wherein, in the above use, ring A is selected from phenyl and pyridyl.

[0046] Preferably, in the above use, the structural unit Selected from

[0047] More preferably, in the above use, the structural unit Selected from

[0048] Wherein, in the above use, the compound of formula (I) is selected from:

[0049] in,

[0050] R1 and R3 are as defined above;

[0051] n and m are independently selected from 0, 1, 2 or 3, and n and m are not 0 at the same time;

[0052] R2 is as defined above;

[0053] L1 and L2 are as defined above.

[0054] Wherein, in the above use, the compound of formula (I) is selected from:

[0055] wherein n, m, R1, R2, R3, L1, and L2 are as defined above.

[0056] Wherein, in the above use, the compound of formula (I) is selected from:

[0057] Wherein, in the above use, the compound of formula (I) is selected from:

[0058] Definition and Explanation:

[0059] Unless otherwise indicated, the following terms and phrases used herein are intended to have the following meanings. A particular term or phrase should not be construed as indefinite or unclear unless specifically defined, but rather should be understood in accordance with its ordinary meaning. When a trade name appears in this document, it is intended to refer to the corresponding commercial product or its active ingredient.

[0060] The term "pharmaceutically acceptable" as used herein refers to those compounds, materials, compositions and / or dosage forms which, within the scope of sound medical judgment, are suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response or other problems or complications, commensurate with a reasonable benefit / risk ratio.

[0061] The term "pharmaceutically acceptable salt" refers to salts of the compounds of the present invention, prepared by reacting the compounds of the present invention with relatively nontoxic acids or bases. When the compounds of the present invention contain relatively acidic functional groups, base addition salts can be obtained by contacting the compounds with a sufficient amount of base in neat solution or in a suitable inert solvent. Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amine, or magnesium salts, or similar salts. When the compounds of the present invention contain relatively basic functional groups, acid addition salts can be obtained by contacting the compounds with a sufficient amount of acid in neat solution or in a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include inorganic acid salts such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, bicarbonate, phosphoric acid, monohydrogen phosphate, dihydrogen phosphate, sulfuric acid, bisulfate, hydroiodic acid, phosphorous acid, and the like; and organic acid salts such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, and methanesulfonic acid; and salts of amino acids (such as arginine) and organic acids such as glucuronic acid. Certain specific compounds of the present invention contain both basic and acidic functional groups and can be converted into either base or acid addition salts.

[0062] Pharmaceutically acceptable salts of the present invention can be synthesized by conventional chemical methods from parent compounds containing acid radicals or bases. Generally, such salts are prepared by reacting these compounds in free acid or base form with a stoichiometric amount of a suitable base or acid in water or an organic solvent or a mixture of the two.

[0063] With respect to a drug or pharmacologically active agent, the term "effective amount" or "therapeutically effective amount" refers to a non-toxic amount of the drug or agent sufficient to achieve the intended effect. For the oral dosage forms of the present invention, an "effective amount" of an active substance in the composition means the amount required to achieve the intended effect when used in combination with another active substance in the composition. The determination of an effective amount varies from person to person, depending on the age and general condition of the recipient, as well as the specific active substance. The appropriate effective amount in each individual case can be determined by those skilled in the art through routine experimentation.

[0064] The terms "active ingredient," "therapeutic agent," "active substance," or "active agent" refer to a chemical entity that is effective in treating a target disorder, disease, or condition.

[0065] The structures of the compounds of the present invention can be confirmed by conventional methods well known to those skilled in the art. If the present invention relates to the absolute configuration of a compound, the absolute configuration can be confirmed by conventional techniques in the art. For example, single crystal X-ray diffraction (SXRD) is used to collect diffraction intensity data from a cultured single crystal using a Bruker D8venture diffractometer, using CuKα radiation and a φ / ω scanning mode. After collecting relevant data, the crystal structure can be further analyzed using a direct method (Shelxs97) to confirm the absolute configuration.

[0066] The compounds of the present invention may exist in specific geometric or stereoisomeric forms. The present invention contemplates all such compounds, including cis- and trans-isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)-isomers, (L)-isomers, and racemic mixtures and other mixtures thereof, such as enantiomerically or diastereomerically enriched mixtures, all of which are within the scope of the present invention. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers and mixtures thereof are encompassed within the scope of the present invention.

[0067] Unless otherwise indicated, the term "enantiomer" or "optical isomer" refers to stereoisomers that are mirror images of one another.

[0068] Unless otherwise indicated, the term "cis-trans isomers" or "geometric isomers" arises from the inability to rotate freely about double bonds or single bonds forming ring carbon atoms.

[0069] Unless otherwise indicated, the term "diastereomer" refers to stereoisomers that have two or more chiral centers and that are not mirror images of each other.

[0070] Unless otherwise indicated, "(+)" indicates dextrorotatory, "(-)" indicates levorotatory, and "(±)" indicates racemic.

[0071] Unless otherwise specified, use a solid wedge key. and dotted wedge key To indicate the absolute configuration of a stereocenter, use a straight solid bond and straight dashed key Indicate the relative configuration of stereocenters with a wavy line Indicates a wedge-shaped solid key or dotted wedge key Or use a wavy line Indicates a straight solid bond or straight dashed key

[0072] Unless otherwise indicated, the terms "enriched in one isomer", "isomerically enriched", "enriched in one enantiomer" or "enantiomerically enriched" mean that the content of one isomer or enantiomer is less than 100%, and the content of that isomer or enantiomer is greater than or equal to 60%, or greater than or equal to 70%, or greater than or equal to 80%, or greater than or equal to 90%, or greater than or equal to 95%, or greater than or equal to 96%, or greater than or equal to 97%, or greater than or equal to 98%, or greater than or equal to 99%, or greater than or equal to 99.5%, or greater than or equal to 99.6%, or greater than or equal to 99.7%, or greater than or equal to 99.8%, or greater than or equal to 99.9%.

[0073] Unless otherwise indicated, the term "isomer excess" or "enantiomeric excess" refers to the difference between the relative percentages of two isomers or two enantiomers. For example, if the content of one isomer or enantiomer is 90% and the content of the other isomer or enantiomer is 10%, the isomer or enantiomeric excess (ee value) is 80%.

[0074] Optically active (R)- and (S)-isomers, as well as D and L isomers, can be prepared by chiral synthesis or chiral reagents or other conventional techniques. If one enantiomer of a compound of the present invention is desired, it can be prepared by asymmetric synthesis or derivatization with a chiral auxiliary, wherein the resulting diastereomeric mixture is separated and the auxiliary group is cleaved to provide the pure desired enantiomer. Alternatively, when the molecule contains a basic functional group (such as an amino group) or an acidic functional group (such as a carboxyl group), a diastereomeric salt is formed with an appropriate optically active acid or base, and then the diastereoisomers are resolved by conventional methods known in the art, and then the pure enantiomer is recovered. In addition, the separation of enantiomers and diastereomers is typically accomplished by using chromatography, which employs a chiral stationary phase and is optionally combined with a chemical derivatization method (e.g., carbamate formation from an amine).

[0075] The compounds of the present invention may contain unnatural proportions of atomic isotopes at one or more of the atoms that constitute the compound. For example, the compounds may be labeled with radioactive isotopes, such as tritium ( 3 H), iodine-125( 125 I) or C-14( 14 C). For example, deuterated drugs can be formed by replacing hydrogen with heavy hydrogen. The bond between deuterium and carbon is stronger than the bond between ordinary hydrogen and carbon. Compared with non-deuterated drugs, deuterated drugs have advantages such as reduced toxic side effects, increased drug stability, enhanced efficacy, and prolonged drug biological half-life. All isotopic variations of the compounds of the present invention, whether radioactive or not, are included within the scope of this invention.

[0076] The terms "optional" or "optionally" mean that the subsequently described event or circumstance may but need not occur, and that the description includes instances where said event or circumstance occurs and instances where it does not.

[0077] The term "substituted" means that any one or more hydrogen atoms on a particular atom are replaced by a substituent, which may include deuterium and hydrogen variants, as long as the valence state of the particular atom is normal and the substituted compound is stable. When the substituent is oxygen (i.e., =O), it means that two hydrogen atoms are replaced. Oxygen substitution does not occur on aromatic groups. The term "optionally substituted" means that it may be substituted or unsubstituted, and unless otherwise specified, the type and number of substituents can be any on the basis of chemical achievable.

[0078] When any variable (e.g., R) occurs more than once in a compound's composition or structure, its definition at each occurrence is independent. Thus, for example, if a group is substituted with 0-2 Rs, the group may be optionally substituted with up to two Rs, with each occurrence of R being an independent choice. Furthermore, combinations of substituents and / or their variants are permissible only if such combinations result in stable compounds.

[0079] When the number of a linking group is 0, such as -(CRR)0-, it means that the linking group is a single bond.

[0080] When the number of a substituent is 0, it means that the substituent does not exist, for example, -A-(R)0 means that the structure is actually -A.

[0081] When a substituent is vacant, it means that the substituent does not exist. For example, when X in AX is vacant, it means that the structure is actually A.

[0082] When one of the variables is selected from a single bond, it means that the two groups it connects are directly connected. For example, when L in ALZ represents a single bond, it means that the structure is actually AZ.

[0083] When a substituent's bond can cross-link to two or more atoms in a ring, the substituent can be bonded to any atom in the ring, e.g. The substituent R can be substituted at any position on the cyclohexyl group or cyclohexadiene. When the listed substituent does not specify the atom through which it is bonded to the substituted group, the substituent can be bonded through any atom. For example, a pyridyl substituent can be bonded to the substituted group through any carbon atom on the pyridine ring.

[0084] When the listed linking groups do not specify their linking direction, their linking direction is arbitrary, for example, The connecting group L is -MW-, in which case -MW- can connect ring A and ring B in the same direction as the reading order from left to right to form You can also connect ring A and ring B in the opposite direction of reading from left to right to form Combinations of linkers, substituents, and / or variations thereof are permissible only if such combinations result in stable compounds.

[0085] Unless otherwise specified, when a group has one or more connectable sites, any one or more sites of the group can be connected to other groups through chemical bonds. When the chemical bond connection mode is non-positional and there are H atoms at the connectable sites, when the chemical bond is connected, the number of H atoms at the site will decrease accordingly with the number of connected chemical bonds, and become a group with a corresponding valence. The chemical bond connecting the site to other groups can be a straight solid bond. Straight dotted key or wavy lines For example, the straight solid bond in -OCH3 indicates that it is connected to other groups through the oxygen atom in the group; The straight dashed bond in the group indicates that the two ends of the nitrogen atom in the group are connected to other groups; The wavy line in the phenyl group indicates that it is connected to other groups through the carbon atoms at positions 1 and 2 in the phenyl group; Indicates that any linkable site on the piperidinyl group can be connected to other groups through a chemical bond, including at least In these four connection methods, even if an H atom is drawn on -N-, Still includes For groups connected in this way, when one chemical bond is connected, the H at that site will be reduced by one and become a corresponding monovalent piperidine group.

[0086] Unless otherwise specified, the number of atoms in a ring is generally defined as the number of members of the ring, for example, a "5-7 membered ring" refers to a "ring" having 5-7 atoms arranged around it.

[0087] Unless otherwise specified, the term “C 1-6 "Alkyl" is used to represent a straight or branched chain saturated hydrocarbon group consisting of 1 to 6 carbon atoms. 1-6 Alkyl groups include C 1-5 、C 1-4 、C 1-3 、C 1-2 、C 2-6 、C 2-4, C6 and C5 alkyl, etc.; which can be monovalent (such as methyl), divalent (such as methylene) or polyvalent (such as methine). 1-6 Examples of alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), butyl (including n-butyl, isobutyl, s-butyl and t-butyl), pentyl (including n-pentyl, isopentyl and neopentyl), hexyl, and the like.

[0088] Unless otherwise specified, the term “C 1-3 "Alkyl" is used to represent a straight or branched chain saturated hydrocarbon group consisting of 1 to 3 carbon atoms. 1-3 Alkyl groups include C 1-2 and C 2-3 Alkyl, etc.; it can be monovalent (such as methyl), divalent (such as methylene) or polyvalent (such as methine). 1-3 Examples of alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), and the like.

[0089]

[0046] The terms "halo" or "halogen," by themselves or as part of another substituent, mean, unless otherwise stated, a fluorine, chlorine, bromine, or iodine atom.

[0090] Unless otherwise specified, “C 3-6 "Cycloalkyl" refers to a saturated cyclic hydrocarbon group consisting of 3 to 6 carbon atoms, which is a monocyclic and bicyclic ring system, wherein the carbon atoms may be optionally oxidized (ie, C=O). 3-6 Cycloalkyl groups include C 3-5 、C 4-5 and C 5-6 Cycloalkyl, etc.; it may be monovalent, divalent or polyvalent. 3-6 Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like.

[0091] Unless otherwise specified, “C 2-6 "Alkenyl" is used to refer to a linear or branched hydrocarbon group consisting of 2 to 6 carbon atoms containing at least one carbon-carbon double bond, which may be located at any position of the group. 2-6 Alkenyl groups include C 2-4 、C 2-3 , C4, C3 and C2 alkenyl, etc.; which may be monovalent, divalent or polyvalent. 2-6 Examples of alkenyl groups include, but are not limited to, ethenyl, propenyl, butenyl, pentenyl, hexenyl, butadienyl, piperyl, hexadienyl, and the like.

[0092] Unless otherwise specified, “C 2-4"Alkenyl" is used to refer to a linear or branched hydrocarbon group consisting of 2 to 4 carbon atoms containing at least one carbon-carbon double bond, which may be located at any position of the group. 2-4 Alkenyl groups include C 2-3 , C4, C3 and C2 alkenyl, etc.; the C 2-4 Alkenyl groups can be monovalent, divalent, or polyvalent. 2-4 Examples of alkenyl groups include, but are not limited to, ethenyl, propenyl, butenyl, butadienyl, and the like.

[0093] Unless otherwise specified, “C 2-6 "Alkynyl" is used to represent a linear or branched hydrocarbon group consisting of 2 to 6 carbon atoms containing at least one carbon-carbon triple bond, which may be located at any position of the group. 2-6 Alkynyl groups include C 2-4 、C 2-3 , C4, C3 and C2 alkynyl, etc. It can be monovalent, divalent or polyvalent. 2-6 Examples of alkynyl groups include, but are not limited to, ethynyl, propynyl, butynyl, pentynyl, and the like.

[0094] Unless otherwise specified, “C 2-4 "Alkynyl" is used to represent a linear or branched hydrocarbon group consisting of 2 to 4 carbon atoms containing at least one carbon-carbon triple bond, which may be located at any position of the group. 2-4 Alkynyl groups include C 2-3 , C4, C3 and C2 alkynyl, etc. It can be monovalent, divalent or polyvalent. 2-4 Examples of alkynyl groups include, but are not limited to, ethynyl, propynyl, butynyl, and the like.

[0095] Unless otherwise specified, the term “C 1-3 "Alkoxy" refers to those alkyl groups containing 1 to 3 carbon atoms which are attached to the rest of the molecule via an oxygen atom. The C 1-3 Alkoxy groups include C 1-2 、C 2-3 , C3 and C2 alkoxy, etc. 1-3 Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy (including n-propoxy and isopropoxy), and the like.

[0096] Unless otherwise specified, the term “C 1-3 "Alkylamino" means an alkyl group containing 1 to 3 carbon atoms which is attached to the rest of the molecule via an amino group. 1-3 Alkylamino groups include C 1-2 , C3 and C2 alkylamino, etc. 1-3Examples of alkylamino groups include, but are not limited to, -NHCH3, -N(CH3)2, -NHCH2CH3, -N(CH3)CH2CH3, -NHCH2CH2CH3, -NHCH2(CH3)2, and the like.

[0097] Unless otherwise specified, the term "3-6 membered heterocycloalkyl" by itself or in combination with other terms means a saturated cyclic group consisting of 3 to 6 ring atoms, 1, 2, 3 or 4 of which are heteroatoms independently selected from O, S and N, and the rest are carbon atoms, wherein the nitrogen atom is optionally quaternized and the nitrogen and sulfur heteroatoms are optionally oxidized (i.e., NO and S(O) p , p is 1 or 2). It includes monocyclic and bicyclic ring systems, wherein bicyclic ring systems include spirocyclic, fused and bridged rings. In addition, with respect to the "3-6 membered heterocycloalkyl", a heteroatom may occupy the position at which the heterocycloalkyl is connected to the rest of the molecule. The 3-6 membered heterocycloalkyl includes 4-6 membered, 5-6 membered, 4 membered, 5 membered and 6 membered heterocycloalkyls, etc. Examples of 3-6 membered heterocycloalkyl groups include, but are not limited to, azetidinyl, oxetanyl, thietanyl, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, tetrahydrothiophenyl (including tetrahydrothiophen-2-yl and tetrahydrothiophen-3-yl, etc.), tetrahydrofuranyl (including tetrahydrofuran-2-yl, etc.), tetrahydropyranyl, piperidinyl (including 1-piperidinyl, 2-piperidinyl and 3-piperidinyl, etc.), piperazinyl (including 1-piperazinyl and 2-piperazinyl, etc.), morpholinyl (including 3-morpholinyl and 4-morpholinyl, etc.), dioxanyl, dithianyl, isoxazolidinyl, isothiazolidinyl, 1,2-oxazinyl, 1,2-thiazinyl, hexahydropyridazinyl, homopiperazinyl or homopiperidinyl, etc.

[0098] Unless otherwise specified, the terms "5-6 membered heteroaromatic ring" and "5-6 membered heteroaryl" are used interchangeably in the present invention. The term "5-6 membered heteroaryl" refers to a monocyclic group with a conjugated π electron system consisting of 5 to 6 ring atoms, wherein 1, 2, 3 or 4 ring atoms are heteroatoms independently selected from O, S and N, and the rest are carbon atoms. The nitrogen atom is optionally quaternized, and the carbon, nitrogen and sulfur heteroatoms are optionally oxidized (i.e., C=O, NO and S(O) p, p is 1 or 2). 5-6 membered heteroaryl can be connected to the rest of the molecule through a heteroatom or a carbon atom. The 5-6 membered heteroaryl includes 5-membered and 6-membered heteroaryl. Examples of the 5-6 membered heteroaryl include, but are not limited to, pyrrolyl (including N-pyrrolyl, 2-pyrrolyl and 3-pyrrolyl, etc.), pyrazolyl (including 2-pyrazolyl and 3-pyrazolyl, etc.), imidazolyl (including N-imidazolyl, 2-imidazolyl, 4-imidazolyl and 5-imidazolyl, etc.), oxazolyl (including 2-oxazolyl, 4-oxazolyl and 5-oxazolyl, etc.), triazolyl (1H-1,2,3-triazolyl, 2H-1,2,3-triazolyl, 1H-1,2,4-triazolyl) and 4H-1,2,4-triazolyl, etc.), tetrazolyl, isoxazolyl (3-isoxazolyl, 4-isoxazolyl and 5-isoxazolyl, etc.), thiazolyl (including 2-thiazolyl, 4-thiazolyl and 5-thiazolyl, etc.), furyl (including 2-furyl and 3-furyl, etc.), thienyl (including 2-thienyl and 3-thienyl, etc.), pyridyl (including 2-pyridyl, 3-pyridyl and 4-pyridyl, etc.), pyrazinyl or pyrimidinyl (including 2-pyrimidinyl and 4-pyrimidinyl, etc.).

[0099] The compounds of the present invention can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining them with other chemical synthesis methods, and equivalent substitutions well known to those skilled in the art. Preferred embodiments include but are not limited to the examples of the present invention.

[0100] The solvent used in the present invention is commercially available.

[0101] The present invention uses the following abbreviations: aq represents water; eq represents equivalent; DCM represents dichloromethane; PE represents petroleum ether; DMF represents N,N-dimethylformamide; DMSO represents dimethyl sulfoxide; EtOAc represents ethyl acetate; EtOH represents ethanol; MeOH represents methanol; Cbz represents benzyloxycarbonyl, which is an amine protecting group; Boc represents tert-butyloxycarbonyl, which is an amine protecting group; HOAc represents acetic acid; THF represents tetrahydrofuran; and LDA represents lithium diisopropylamide.

[0102] Compounds are named according to the conventional nomenclature in the art or using Software naming, commercially available compounds use supplier catalog names.

[0103] Note: The compounds of formula (I) of the present invention can refer to the general formula of the compounds and related specific compounds in WO2021 / 057893A1. Beneficial effects:

[0104] The compounds of the present invention have high activity and high selectivity for BTK kinase, and have significant therapeutic effects on BTK-related diseases such as lymphoma, leukemia, autoimmune diseases, inflammatory diseases or metabolic diseases, and have synergistic and enhancing effects with many types of compounds and drugs with different mechanisms; in addition, the compounds of the present invention can effectively penetrate the blood-brain barrier and maintain high concentrations in brain tissue, so they can achieve therapeutic effects on diseases of the central nervous system; after efficacy tests, the compounds of the present invention have demonstrated significant effects in the treatment of central nervous system diseases such as multiple sclerosis and central nervous system lymphoma, and are expected to be used as a new method for treating such central nervous system diseases. BRIEF DESCRIPTION OF THE DRAWINGS

[0105] FIG1 shows the clinical scores of mice with autoimmune encephalomyelitis induced by MOG35-55 in Experimental Example 2.

[0106] FIG2 is a graph of brain luminescence intensity during in vivo imaging of the Namalwa intracranial model experiment in Experimental Example 4. DETAILED DESCRIPTION

[0107] Example 1

[0108] The synthetic route is as follows:

[0109] Step 1. Synthesis of I-3

[0110] Compound I-1 (3 kg, 23.06 mol) was dissolved in toluene (5 L). A 50% (mass fraction) aqueous NaOH solution (1.84 kg, 23.06 mol) was added with stirring. The reaction was heated to 135°C. Water was separated using an oil-water separator during the reaction. The reaction was continued for a total of 96 hours. The reaction solution was transferred to a 3 L single-necked flask and concentrated to dryness under reduced pressure. The concentrate was transferred to a reaction flask, and compound I-2 (6.00 kg, 25.43 mol) and DMF (2.5 L) were added. The reaction system was heated to 145°C to melt. Cu (147.00 g, 2.31 mol) and CuBr (330.00 g, 2.30 mol) were added and the reaction was maintained at 145°C for 64 hours. The heating and stirring were turned off, the reaction solution was cooled to room temperature, and the solids were allowed to settle. The supernatant was then decanted. 1.5 L of n-heptane was added and stirred for 5 minutes. The supernatant was again decanted. This operation was repeated four times. Combine all supernatants and transfer to a 50L separator, wash with a mixed solution of 2.5L ammonia (content 25% to 28%) and 2.5L water, repeat 5 times, then wash with 10L semi-saturated brine, collect the organic phase, combine the aqueous phase, extract the aqueous phase with 10L n-heptane, combine the above two organic phases, dry over anhydrous sodium sulfate, filter out the supernatant, and concentrate to dryness under reduced pressure. The concentrate is subjected to reduced pressure distillation in batches (external temperature 170°C), first preheat the concentrate to 150°C for 1 hour, and then perform reduced pressure distillation with a water pump and then an oil pump in sequence, discard the distilled fraction, and collect the residue. The residue is purified by silica gel column to obtain compound I-3.

[0111] 1 H NMR (400MHz, DMSO-d6) δppm 7.56 (d, J = 8.78Hz, 2H), 7.17-7.37 (m, 2H), 6.90-7.11 (m, 3H).

[0112] Step 2. Synthesis of I-5

[0113] DCM (3.2 L) was added to the reaction flask, followed by compound I-4 (800 g, 3.72 mol) and TEMPO (5.97 g, 37.94 mmol). KBr aqueous solution (2 M, 192.00 mL) was added to the resulting solution, which was then transferred to an ice bath. NaHCO3 (44.00 g, 523.75 mmol) was added, and the temperature was lowered to below 10°C, followed by addition of NaClO (4.84 kg, 5.20 mol, 4.00 L, 8% purity) at a temperature maintained at 10-40°C. The temperature was then added dropwise for about 30 minutes. After the addition was complete, the temperature was naturally lowered to 30°C and the reaction was continued for 10 minutes. The reaction solution was separated, the aqueous phase was extracted with 3.2 L of dichloromethane, and the combined organic phases were washed successively with 1.6 L of hydrochloric acid (1 M, containing KI (616.85 g, 3.72 mol) and 1.6 L of sodium thiosulfate solution (10%). The obtained organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to dryness under reduced pressure to give compound I-5.

[0114] 1H NMR (400MHz, CDCl3) δppm 9.68(s,1H),3.79-4.02(m,1H),3.55-3.71(m,1H),3.25-3.39(m,1H),3.01-3.15(m ,1H),2.34-2.48(m,1H),1.87-2.01(m,1H),1.61-1.71(m,2H),1.41-1.49(m,10H).

[0115] Step 3. Synthesis of I-7

[0116] Compound I-6 (1.68 kg, 14.60 mol) was dissolved in THF (11.25 L) and added to a 50 L reactor. Cooled to -73 to -63 ° C using a dry ice ethanol bath. Nitrogen was passed through, and n-BuLi (2.5 M, 6.09 L) was added to the reactor via a peristaltic pump under a weak nitrogen flow, maintaining the temperature below -50 ° C. After the addition was complete, the temperature was maintained at -68 to -50 ° C for 1 hour. Compound I-5 (3 kg, 14.07 mol) was dissolved in THF (3.75 L) and added to the reactor via a peristaltic pump, maintaining the temperature below -50 ° C. After the addition was complete, the dry ice ethanol bath was removed and the reaction was slowly heated to 0 ° C for 2 hours. 10 L of saturated ammonium chloride solution was added dropwise to the reaction solution, and the mixture was stirred for 0.5 hours and then concentrated. After concentrating until ~15 L of solution remained, the mixture was transferred to a separator, 10 L of hydrochloric acid (1 M) was added, and the mixture was extracted with ethyl acetate (10 L*2). The combined organic phases were washed with 10 L of semi-saturated brine, and the obtained organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain I-7.

[0117] LCMS (ESI) m / z: 273.1 (M-56); 1H NMR (400MHz, CDCl3) δppm 8.25-8.38(m,2H),4.79-4.91(m,1H),4.01-4.15(m,0.5H),3.79-3.96(m,1H),3.66- 3.74(m,0.5H),3.38-3.54(m,1H),2.75-2.87(m,0.5H),2.50-2.65(m,0.5H),2.15(br d,J=12.05Hz,1H),1.98-2.08(m,1H),1.54-1.80(m,2H),1.42-1.52(m,10H).

[0118] Step 4. Synthesis of I-8

[0119] A solution of compound I-7 (4 kg, 12.18 mol) and TEMPO (19.20 g, 122.10 mmol) in DCM (10 L) was added to a reactor, followed by addition of KBr (2 M, 630 mL) (aqueous solution) and NaHCO 3 (144 g, 1.71 mol, 66.67 mL). NaClO (15.87 kg, 17.05 mol, 13.12 L, 8% purity) was added using a peristaltic pump while maintaining the temperature at 10-30° C. The reaction was maintained at 25-30° C. for 30 min. The reaction solution was separated, the aqueous phase was extracted with 10 L of dichloromethane, and the combined organic phases were washed sequentially with 10 L of hydrochloric acid (1 M, containing 2 kg of KI) and 10 L of sodium thiosulfate solution (10%). The resulting organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give compound I-8.

[0120] LCMS (ESI) m / z: 271.1 (M-56); 1 H NMR (400MHz, CDCl3) δppm 8.38-8.50(m,2H),4.02-4.20(m,1H),3.73-3.86(m,1H),3.08-3.25(m,1H ),2.88-3.04(m,2H),1.91-2.03(m,1H),1.68-1.85(m,2H),1.41(s,10H).

[0121] Step 5. Synthesis of I-9

[0122] To a solution of compound I-8 (5kg, 15.32mol) in dioxane (25L) was added hydrazine hydrate (1kg, 16.98mol, 970.87mL, 85% content), and the temperature was raised to 93°C for 16 hours. After cooling to room temperature, the reaction solution was concentrated under reduced pressure. The concentrate was dissolved in 15L of dichloromethane and added to a separatory vessel. 5L of water was added and the pH was adjusted to about 8 with a 1M NaOH solution. The liquid was separated and the organic phase was collected. The organic phase was dried over anhydrous sodium sulfate, filtered, and the filter cake was washed with 0.5L of dichloromethane. The combined filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was dispersed in 6L of ethyl acetate, 18L of n-heptane was added under stirring, and the mixture was stirred at room temperature for 3 hours before filtering. The filter cake was washed with 4L of a mixed solvent (n-heptane: ethyl acetate = 3:1). The filter cake was filtered to dryness, the filter cake was collected, and the combined filtrate was concentrated under reduced pressure to dryness. The concentrate was purified via a silica gel column (n-heptane:ethyl acetate = 1:0 to 1:1, with 20% dichloromethane added). The resulting product fraction was concentrated to dryness under reduced pressure. The concentrate and the filter cake were dispersed in n-heptane (8 L), stirred at room temperature (20°C) for 1 hour, and then filtered. The filter cake was filtered with an oil pump to dryness and dried in a vacuum drying oven at 50°C for 16 hours. Compound I-9 was obtained.

[0123] LCMS (ESI) m / z: 321.2 (M+1); 1 H NMR (400MHz, CDCl3) δppm 11.95-12.70(m,1 H),8.79(br s,1H),8.09(br s,1H),4.23-4.61(m,1H),4.11(br d,J=13.05Hz,1H),3.13-3.45(m,2H),2.90(br t,J=11.42Hz,1H),2.20(br d,J=10.54Hz,1H),1.84(br s,2H),1.60-1.71(m,1H),1.37-1.59(m,9H).

[0124] Step 6. Synthesis of I-10

[0125] 16L of anhydrous dioxane, compound I-3 (3033g, 10.37mol), compound I-9 (3250g, 10.14mol), N,N'-dimethylethylenediamine (482.5g, 5.47mol), cuprous iodide (482.5g, 2.53mol), and potassium phosphate (3900g, 18.37mol) were added to the reactor in sequence, the reaction temperature was adjusted to 95-105°C, and stirred for 16 hours. The reaction liquid temperature was lowered to 15-25°C, the reaction liquid was filtered through diatomaceous earth, the filter cake was washed with dichloromethane (1.6L*3), and the filtrate was concentrated under reduced pressure. The solid was dissolved in 16L of dichloromethane, transferred to a separatory vessel, and washed with aqueous ammonia (mass fraction 25%-28%) (1.6L*3) and 16L of semi-saturated brine in sequence. The organic phase was dried over anhydrous sodium sulfate, filtered, and the filter cake was washed with 1.6 L of dichloromethane. The combined filtrates were concentrated under reduced pressure to obtain a crude product. The crude product was dispersed in 32 L of a mixed solvent (n-heptane: ethyl acetate = 7:1), stirred at room temperature of 10-20 ° C for 16 hours, filtered, and the filter cake was washed with a mixed solvent (n-heptane: ethyl acetate = 7:1, 3.2 L * 3), and the filter cake was collected. The filter cake was dispersed in 20.8 L of a mixed solvent (n-heptane: ethyl acetate = 7:1), stirred at room temperature of 10-20 ° C for 16 hours, filtered, and the filter cake was washed with a mixed solvent (n-heptane: ethyl acetate = 7:1, 2.08 L * 3), and the filter cake was collected and dried under vacuum at 55-65 ° C to constant weight to obtain compound I-10.

[0126] LCMS (ESI) m / z: 525.2 (M+1); 1 H NMR(400MHz, CDCl3)δppm 8.95(s,1H),8.20(s,1H),7.60-7.80(m,2H),7.14-7.24(m,2H),6.98-7.13(m,2H),6.86-6.94(m,1H),4.39(br s,1H),4.12(br d,J=12.30Hz,1H),3.31-3.43(m,1H),3.21(br t,J=11.17Hz,1H),2.83-2.98(m,1H),2.26(br d,J=13.05Hz,1H),1.89(br d,J=10.29Hz,2H),1.65-1.75(m,1H),1.44(br s,9H).

[0127] Step 7. Synthesis of I-11

[0128] A 4M hydrogen chloride / dioxane solution (16.4 L) was added to the reactor, and compound I-10 (4100 g, 7.81 mol) was added in batches at an internal temperature of 10-30°C. The reaction mixture was stirred for 0.5 hours while maintaining the internal temperature at 10-30°C. The reaction solution was concentrated to dryness under reduced pressure, and 8.2 L of dichloromethane was added and further concentrated to dryness under reduced pressure. The resulting oil was dissolved in 12.3 L of water and the pH was adjusted to 8-9 with solid sodium carbonate. Stirring was continued for 0.5 hours, and then the mixture was extracted with 16.4 L of dichloromethane. The organic phase was washed with 8.2 L of water and the organic phase was collected. The resulting aqueous phase was extracted with 8.2 L of dichloromethane, and the organic phase was collected. The two organic phases collected above were combined. The mixed organic phase was washed with 12.3 L of semi-saturated brine. The organic phase was dried over anhydrous sodium sulfate, filtered, and the filter cake was washed with 2.05 L of dichloromethane. The combined filtrate was concentrated to dryness under reduced pressure. Supercritical fluid chromatography (SCFC) (Chiralpak IG-350 × 4.6 mm ID, 3 μm column; mobile phase: A: supercritical carbon dioxide, B: 0.05% diethylamine in ethanol; gradient: B from 5% to 40% over 2 minutes, to 5% over 0.5 minutes, 40% maintained for 1.2 minutes, 5% maintained for 0.8 minutes; flow rate: 4 mL / min; column temperature: 35°C; wavelength: 220 nm) confirmed the racemic compound. SFC separation (DAICEL CHIRALPAK AD column (250 mm x 50 mm, 10 μm); mobile phase: A: supercritical carbon dioxide, B: [0.1% NH3H2O ​​MeOH]; B%: 50%-50%) was performed. The solid obtained after separation was dissolved in 16.4 L of dichloromethane, washed with water (8.2 L x 2), and the organic phase was collected. The aqueous phases from the two washes were combined and extracted with 8.2 L of dichloromethane. The organic phase was collected, the two organic phases collected above were combined, and washed with 16.4 L of semi-saturated brine. The organic phase was dried over anhydrous sodium sulfate and filtered. The filter cake was washed with 2.05 L of dichloromethane. The combined filtrate was concentrated to dryness under reduced pressure to obtain compound I-11. Supercritical fluid chromatography (Chromatographic column: Chiralpak IG-3 50×4.6 mm ID, 3 μm; Mobile phase: A: supercritical carbon dioxide, B: 0.05% diethylamine in ethanol; Gradient: B from 5% to 40% over 2 minutes, from 5% to 5% over 0.5 minutes, 40% maintained for 1.2 minutes, 5% maintained for 0.8 minutes; Flow rate: 4 mL / min; Column temperature: 35°C; Wavelength: 220 nm) revealed a single-configuration compound with a retention time of 2.584 minutes.

[0129] LCMS (ESI) m / z: 425.2 (M+1); 1H NMR (400MHz, DMSO-d6) δppm 9.18-10.35(m,1H),9.02-9.15(m,1H),8.33(s,1H),7.86(d,J=8.88Hz,2H),7.19-7.45(m,4H),7.09(br t,J=7.57Hz,1H),3.69-3.84(m,1H),3.61(br d,J=12.01Hz,1H),3.18-3.38(m,2H),2.88-3.02(m,1H),2.22(br d,J=11.26Hz,1H),1.88-2.03(m,2H),1.72-1.88(m,1H).

[0130] Step 8. Synthesis of Compound 1A

[0131] Set up the redistillation device, add compound I-12 (500g) to the reaction bottle, raise the external temperature to 90-110℃ and start redistillation, discard the fraction with a steam temperature of less than 40℃, collect the fraction with a steam temperature of 40-80℃, and collect until about 15mL is collected at the bottom of the bottle. Transfer the collected fraction to the redistillation device for a second redistillation, discard the fraction with a steam temperature of less than 40℃, collect the fraction with a steam temperature of 40-80℃, and collect until about 15mL is collected at the bottom of the bottle. Transfer the collected fraction to the redistillation device for a third redistillation, discard the fraction with a steam temperature of less than 40℃, collect the fraction with a steam temperature of 40-80℃, and collect until about 15mL is collected at the bottom of the bottle. The obtained fraction is set aside. Compound I-11 (1600 g, 3.56 mol) was dissolved in a mixture of tetrahydrofuran (16 L) and water (8 L). This solution was added to a reactor. Potassium phosphate (1513 g, 7.13 mol) was added to the reactor and the internal temperature was lowered to 5-15°C. Re-distilled I-12 (322 g, 3.56 mol) was dissolved in tetrahydrofuran (3.2 L) and the resulting solution was added dropwise to the reactor, maintaining the internal temperature at 5-15°C for 30 minutes. After the addition was complete, stirring was continued at 5-15°C for 10 minutes. The reaction mixture was separated, and the organic and aqueous phases were collected. The aqueous phase was extracted with ethyl acetate (8 L x 2). The organic phase was collected and the two collected organic phases were combined. The combined organic phases were washed sequentially with 8 L of semi-saturated citric acid solution, 8 L of saturated sodium carbonate solution, and 8 L of semi-saturated brine. The organic phase was dried over anhydrous sodium sulfate and filtered. The filter cake was washed with 1.6 L of ethyl acetate, and the combined filtrates were concentrated to dryness under reduced pressure. The resulting oil was dissolved in 16 L of ethyl acetate, and potassium carbonate (493.05 g, 3.56 mol) was added. Stirring was continued at room temperature (10-20°C) for 16 hours. 8 L of water was added and stirring was continued for 10 minutes. After stirring was stopped, the liquids were separated and the organic phase was collected and washed with 8 L of semi-saturated brine. The organic phase was dried over anhydrous sodium sulfate and filtered. The filter cake was washed with 1.6 L of ethyl acetate, and the combined filtrates were concentrated to approximately 4 L under reduced pressure. 1600 g of 200-300 mesh silica gel was placed in a Buchner funnel and moistened with 1.6 L of ethyl acetate. The concentrated solution was filtered, the filter cake was washed with 8 L of ethyl acetate, and the combined filtrates were filtered through a Buchner funnel and concentrated to dryness under reduced pressure. Dissolve the concentrated oil in 3.2 L of ethanol at 55-65°C. After cooling to room temperature (10-20°C), slowly add 4.8 L of water dropwise to the ethanol solution. Oil precipitates. Add 100 mg of seed crystals and adjust the stirring speed to 400 rpm. Solids gradually precipitate. Continue stirring at room temperature for 16 hours. Filter, wash the filter cake with 1.6 L of a mixed solvent (ethanol:water = 2:3), collect the filter cake, and vacuum dry it at 75-85°C for 16 hours. Pass the dried crude product through a 24-mesh sieve.

[0132] The resulting powder was dispersed in 8.8 L of a mixed solvent (n-heptane:ethyl acetate = 10:1), heated to 50-60°C, stirred for 1 hour, then cooled to 25-35°C with stirring. Filtered, the filter cake was washed with 1.76 L of a mixed solvent (n-heptane:ethyl acetate = 10:1), collected, and the above beating process repeated. The resulting filter cake was vacuum-dried at 65-75°C to constant weight to yield compound 1A. Supercritical fluid chromatography (column: Cellulose 2 150×4.6 mm ID, 5 μm; mobile phase: A: supercritical carbon dioxide, B: 0.05% diethylamine in ethanol; gradient: B from 5% to 40% over 5 minutes, 40% hold for 2.5 minutes, return to 5% for 2.5 minutes; flow rate: 2.5 mL / min; column temperature: 35°C; wavelength: 220 nm) revealed a single-configuration compound with a retention time of 6.916 minutes.

[0133] LCMS (ESI) m / z: 479.2 (M+1); 1H NMR (400MHz, DMSO-d6) δppm 9.09 (br s, 1H), 8.32 (br s, 1H), 7.85 (br d, J = 8.78Hz, 2H), 7.19-7.44 (m, 4H), 7.09 (br t,J=7.65Hz,1H),6.75-6.96(m,1H),6.09(br t,J=16.19Hz,1H),5.53-5.77(m,1H),3.99-4.80(m,2H),3.33-3.55(m,1H),3.13-3.30(m,1H),2.89-3.12(m,1H),2.22(br d,J=11.29Hz,1H),1.74-2.09(m,2H),1.46-1.66(m,1H).

[0134] Configuration confirmation of compound 1A:

[0135] Compound 1A (100 mg, 209.00 μmol) was dissolved in DCM (0.3 mL) at room temperature, and n-heptane (1.5 mL) was added dropwise with stirring. A viscous substance formed, which remained insoluble upon heating (40-50°C). After cooling to room temperature and allowing to stand overnight, a yellow oil formed. After further standing at room temperature in the dark for seven months, transparent crystals appeared in the system. Single crystal diffraction analysis confirmed the aforementioned configuration.

[0136] Single crystal diffraction test:

[0137] Instrument model: Single crystal X-ray diffractometer (SC-XRD) (D8VENTURE)

[0138] Conclusion: The molecular formula of compound 1A is C 26H 21 F3N4O2, Monoclinic crystal system, space group C2, unit cell parameters α=γ=90°, β=90.4900(10)°, volume The absolute configuration parameter Flack value is 0.04(3).

[0139] Biological test data

[0140] Experimental Example 1:

[0141] Purpose of the experiment:

[0142] Study the distribution of the test compound in brain tissue.

[0143] Experimental Materials:

[0144] Sprague Dawley rats (female, 200-300 g, 6-9 weeks old, Beijing Weitonglihua) were used.

[0145] Experimental operation:

[0146] The pharmacokinetic profiles of the compounds following oral administration were tested using a standard protocol. Candidate compounds were prepared as a nearly clear dosing solution at a concentration of 1 mg / mL and administered orally to rats (dose 10 mg / kg). The oral vehicle consisted of a mixture of 5% DMSO, 10% solutol HS15, 20% PEG400, and 65% water. Whole blood, brain, and cerebrospinal fluid were collected at 0.5, 1, and 6 hours post-dose. Whole blood samples were centrifuged at 3200 g for 10 minutes, and the supernatant was separated to obtain plasma samples. Proteins were precipitated by adding a defined volume of acetonitrile containing an internal standard, and the supernatant was centrifuged and injected for quantitative analysis of plasma drug concentrations by LC-MS / MS. Brain tissue was collected by perfusion and homogenized using a high-throughput tissue homogenizer at a 4x homogenization ratio on wet ice. The homogenate was maintained at -60°C or colder until quantitative analysis of brain tissue concentrations by LC-MS / MS. Cerebrospinal fluid (CSF) was collected from animals under isoflurane anesthesia, with 50–100 μL of CSF collected at each sampling time point. The collected CSF was stored at −60°C or colder until quantification by LC-MS / MS. Drug concentrations, plasma-to-brain tissue ratios, and plasma-to-CSF ratios were calculated for the three matrices at different sampling time points. The results are shown in Table 1.

[0147] Table 1. Distribution of drugs in the brain tissue of female rats at different time points after oral administration

[0148] Experimental conclusion: Compound 1A can effectively penetrate the blood-brain barrier and can be distributed in the brain tissue after oral administration. The concentration in the brain tissue is as high as 835 ng / g, so it is expected to achieve a therapeutic effect on diseases of the central nervous system.

[0149] Experimental Example 2:

[0150] Purpose of the experiment:

[0151] This study evaluated the efficacy of the test compound in a mouse model of experimental autoimmune encephalomyelitis (EAE). EAE is a disease model primarily mediated by specifically sensitized CD4+ T cells, using myelin protein immunization in experimental animals. This model shares many clinical, biochemical, immunological, and pathological characteristics with human MS and is currently recognized internationally as an ideal animal model for MS.

[0152] Experimental Materials:

[0153] C57BL / 6 mice (female, 16-18 g, Shanghai Jihui Experimental Animal Breeding Co., Ltd.)

[0154] Experimental operation:

[0155] MOG was injected subcutaneously on Day 0 of the experiment. 35-55 The EAE model was induced in mice and the mice were divided into groups for drug administration. On Day 12 after immunization, the mice began to show clinical symptoms of autoimmune encephalomyelitis. The average clinical score of the vehicle control group gradually increased in the early stage of the experiment, reaching 3.11 points on Day 20, indicating that MOG 35-55 The success of the induced autoimmune encephalomyelitis model in MOG 35-55 On day 0 after immunization, the mice were randomly divided into 5 groups according to body weight, namely vehicle control group, dexamethasone (Dex) group, compound 1A 20 mg / kg group, compound 1A 40 mg / kg group and compound 1A 60 mg / kg group. 35-55 The mice in the modeling group served as the normal control group. This experiment started from Day 0 of the mouse immune modeling and continued to administer the drug for 26 days. The efficacy was evaluated by indicators such as EAE clinical scores.

[0156] The experimental results showed that the mice in the vehicle control group 35-55 Severe EAE clinical symptoms developed from Day 12 to Day 26 after immunization, demonstrating successful EAE model establishment. Compound 1A administered continuously for 26 days significantly improved clinical symptoms in EAE mice. The experimental results are shown in Figure 1.

[0157] Experimental conclusion: Continuous administration of the compound of the present invention for 26 days has a significant therapeutic effect on autoimmune encephalomyelitis model mice; the compound of the present invention has shown a significant effect in the treatment of multiple sclerosis and is expected to be used as a new treatment method for multiple sclerosis.

[0158] Experimental Example 3:

[0159] Purpose of the experiment:

[0160] This study used the OCI-LY10 cell subcutaneous xenograft SCID mouse model to evaluate the anti-tumor effect of the compound.

[0161] Experimental operation:

[0162] (1) Cell culture:

[0163] Human diffuse large B-cell lymphoma OCI-LY10 cells were cultured in IMDM medium containing 20% ​​FBS and maintained in a humidified incubator at 37°C with 5% CO2.

[0164] (2) Tumor cell inoculation:

[0165] Collect OCI-LY10 cells in the logarithmic growth phase, resuspend in IMDM basal medium, add Matrigel at a 1:1 ratio, and adjust the cell concentration to 4×10 7 Under sterile conditions, 0.1 mL of cell suspension was inoculated subcutaneously on the right side of the back of SCID mice at a concentration of 4 × 10 6 / 0.1mL / mouse.

[0166] (3) Preparation of test substance:

[0167] Test compound group: weigh a certain amount of the test compound into a brown dispensing bottle, add the corresponding volume of solvent and vortex to obtain a uniform suspension or clear solution.

[0168] Tumor diameter was measured with a vernier caliper twice a week. Tumor volume was calculated using the formula: 1 / 2×a×b 2 , where a and b are the length and width of the tumor, respectively.

[0169] The tumor inhibition efficacy of the compound was evaluated using the TGI (%). The tumor inhibition rate (TGI) (%) was calculated as follows: TGI (%) = [1 - (average tumor volume of a treatment group at the end of dosing - average tumor volume of the treatment group at the start of dosing) / (average tumor volume of the solvent control group at the end of treatment - average tumor volume of the solvent control group at the start of treatment)] × 100%.

[0170] Statistical analysis:

[0171] In this study, experimental data were expressed as Mean ± SEM.

[0172] Statistical analysis was performed using IBM SPSS Statistics software based on RTV data at the end of the experiment. Comparisons between two groups were analyzed using the t test, and comparisons between three or more groups were analyzed using one-way ANOVA. If the variances were equal (F values ​​were not significantly different), Tukey's method was used for analysis; if the variances were unequal (F values ​​were significantly different), the Games-Howell method was used. A p value < 0.05 was considered significant. The results are shown in Table 2.

[0173] Table 2 Effects of the compounds of the present invention on OCI-LY10 cell subcutaneous xenograft tumor model

[0174] Conclusion: Compound 1A of the present invention exhibited significant tumor inhibition in the OCI-LY10 subcutaneous xenograft tumor model, with a clear dose-effect relationship. However, as shown in Experimental Example 1, the compound can penetrate the blood-brain barrier and maintain high concentrations in brain tissue. Combined with the results of Experimental Examples 1 and 3, this suggests that the compound of the present invention can treat central nervous system lymphoma.

[0175] Experimental Example 4

[0176] Namalwa intracranial model experimental protocol: 500,000 luciferase-expressing Namalwa cells were injected via the carotid artery to establish a model in nude mice. Three days after model establishment, mice were randomly divided into a vehicle control group and a Compound 1A-treated group. Treatment began via oral gavage, with Compound 1A administered once daily. The dose of Compound 1A was 150 mg / kg. In vivo imaging was performed 7, 14, and 19 days after model establishment (Figure 2).

[0177] Experimental Results: No brain signals were observed on day 7. Brain tumor signals in the vehicle control group were significantly higher than those in the Compound 1A-treated group on days 14 and 19. There was no statistical difference between the two groups on day 14, but a statistical difference was observed on day 19 (p = 0.035). Rats in the vehicle control group died starting on day 21, so no follow-up data are available.

[0178] Experimental conclusion:

[0179] After treatment, the Compound 1A-treated group showed significantly stronger inhibition of human Burkitt's lymphoma cells in a model of central nervous system Burkitt's lymphoma than the solvent-treated group. The compounds of the present invention have demonstrated efficacy in the treatment of Burkitt's lymphoma and are expected to be used as a new treatment for Burkitt's lymphoma.

Claims

1. Use of a compound represented by formula (I), an isomer thereof, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof in the preparation of a medicament for preventing or treating BTK-related diseases, characterized in that: The BTK-related diseases include lymphoma, leukemia, autoimmune diseases, inflammatory diseases or metabolic diseases; The structure of the compound of formula (I) is as follows: in, T1 is independently selected from N and CH; R2 is independently selected from H, C 1-6 Alkyl, C 2-6 Alkenyl and C 2-6 Alkynyl, the C 1-6 Alkyl, C 2-6 Alkenyl and C 2-6 Alkynyl is optionally substituted by 1, 2 or 3 R a replace; Ring A is selected from phenyl and 5-6 membered heteroaryl; M is independently selected from C 3-6 Cycloalkyl and 3-6 membered heterocycloalkyl, the C 3-6 Cycloalkyl and 3-6 membered heterocycloalkyl are each independently optionally substituted by 1, 2 or 3 R b replace; R1 and R3 are independently selected from F, Cl, Br, I, OH, NH2, CN; n and m are independently selected from 0, 1, 2 or 3, and n and m are not 0 at the same time; L1 and L2 are independently selected from -CH2-, -CH2CH2-, -O-, -C(=O)- and -C(=O)-NH-; R a Independently selected from H, F, Cl, Br, I, OH, NH2, CN, C 1-3 Alkyl, C 1-3 Alkoxy and C 1-3 Alkylamino, the C 1-3 Alkyl, C 1-3 Alkoxy and C 1-3 Alkylamino is optionally substituted with 1, 2 or 3 R groups independently; R b Selected from F, Cl, Br, I, CH3; R is selected from H, F, Cl, Br, I; The 5-6 membered heteroaryl and 3-6 membered heterocycloalkyl groups each independently contain 1, 2, 3 or 4 heteroatoms or heteroatom groups independently selected from -NH-, -O-, -S-, -C(=O)-, -S(=O)-, and N.

2. The use according to claim 1, characterized in that: The BTK-related diseases include central nervous system diseases and non-central nervous system diseases.

3. The use according to claim 2, characterized in that: The central nervous system diseases include central nervous system lymphoma, diffuse large B-cell lymphoma of the central nervous system, Burkitt lymphoma of the central nervous system, autoimmune encephalitis, multiple sclerosis or neuromyelitis optica spectrum disease; the non-central nervous system diseases include B-cell lymphoma, chronic lymphocytic leukemia / small lymphocytic lymphoma, mantle cell lymphoma, Waldenstrom's macroglobulinemia, marginal zone lymphoma, follicular lymphoma, and lymphoma arising from the collar bone, craniofacial bones or abdominal organs. Burkitt lymphoma, non-Hodgkin lymphoma, diffuse large B-cell lymphoma, multiple myeloma, rheumatoid arthritis, systemic lupus erythematosus, lupus nephritis, Sjögren's syndrome, IgG4-related disease, chronic spontaneous urticaria, idiopathic thrombocytopenic purpura, immune thrombocytopenia, pemphigus, asthma, graft-versus-host disease, scleroderma, novel coronavirus infection, granulomatosis with polyangiitis, idiopathic pulmonary fibrosis, psoriasis, atherosclerosis, or diabetes mellitus.

4. The use according to any one of claims 1 to 3, characterized in that: The compound represented by formula (I), its isomer, its pharmaceutically acceptable salt or its pharmaceutical composition is combined with at least one second drug to prepare a drug for preventing or treating BTK-related diseases; the second drug is selected from CD-20 inhibitors or antibodies, BCL-2 inhibitors, SYK inhibitors, PI3K inhibitors, immunosuppressants, chemotherapy drugs or autoimmune checkpoint inhibitors.

5. The use according to claim 4, characterized in that: The mass ratio of the compound represented by formula (I) to the second drug is 1-100:100-1; preferably 1-10:10-1.

6. The use according to claim 4, characterized in that: At least one of the following must be met: The CD-20 inhibitor or antibody includes rituximab, utoximab, ocrelizumab, ibritumomab tiuxetan, obinutuzumab, ofatumumab or vedolizumab; The BCL-2 inhibitors include venetoclax, APG-2575, BGB-11417, ICP-248, FCN-338 or LP-108; The SYK inhibitors include fostamatinib, entospletinib, PRT-2761, HMPL-523, SKI-O-703, IC-265 or GSK-2646264; The PI3K inhibitors include copanlisib, erbulisib, durensibu, adranib, alpelisib, parsaclisib, limplisib, eganelisib, BAY 10-82439, buparlisib, dactolisib, bifentinib, or praquintinib; The immunosuppressants include lenalidomide, pomalidomide, and thalidomide; The chemotherapy drugs include temozolomide, cyclophosphamide, chlorambucil, bendamustine, ifosfamide, prednisone, dexamethasone, cisplatin, carboplatin, oxaliplatin, fludarabine, pentostatin, cladribine, cytarabine, gemcitabine, methotrexate, pralatrexate, anthracyclines, doxorubicin, liposomal doxorubicin, vincristine, mitoxantrone, etoposide or bleomycin; The autoimmune checkpoint inhibitors include pembrolizumab, nivolumab, carrelizumab, toripalimab, cemiplimab, durvalumab, velumab, sintilimab, tislelizumab, sepalimab, durvalumab, atezolizumab, envolimab, sugemalimab, sluglizumab, penampalimab, ceruletumab, zimbalimab, gepnolimab, badalizumab, proglitumab, pucitatinib, cosibelimab, AUNP-12, reluzumab, retivarizumab, dotalimab, cadonimab, socamuzumab, adebrizumab, ipilimumab or tremelimumab.

7. The use according to claim 1, characterized in that: R a Independently selected from H, F, Cl, Br, I, OH, NH2, CN, CH3, OCH3, NH(CH3)2 and N(CH3)2.

8. The use according to claim 1, characterized in that: R2 is independently selected from H, C 1-3 Alkyl, C 2-4 Alkenyl and C 2-4 Alkynyl, the C 1-3 Alkyl, C 2-4 Alkenyl and C 2-4 Alkynyl is optionally substituted by 1, 2 or 3 R a Substituted; preferably, R2 is independently selected from H, CH3, vinyl and propynyl.

9. The use according to claim 1, characterized in that: M is independently selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, azetidinyl, oxetanyl, piperidinyl and morpholinyl, and the cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, azetidinyl, oxetanyl, piperidinyl and morpholinyl are each independently optionally substituted by 1, 2 or 3 R b Substituted; preferably, M is independently selected from piperidinyl and morpholinyl.

10. The use according to claim 1, characterized in that: At least one of the following is satisfied: L1 is selected from -O- and -C(=O)-NH-; L2 is selected from -C(=O)- and -C(=O)-NH-.

11. The use according to claim 1, characterized in that: Ring A is selected from phenyl and pyridyl; preferably, the structural unit Selected from More preferably, the structural unit Selected from 12. The use according to any one of claims 1 to 11, characterized in that: The compound of formula (I) is selected from: in, R1, R3 are as defined in claim 1; n and m are independently selected from 0, 1, 2 or 3, and n and m are not 0 at the same time; R2 is as defined in any one of claims 1 or 5; L1, L2 are as defined in claim 1 or 7.

13. The use according to claim 12, characterized in that: The compound of formula (I) is selected from: wherein n, m, R1, R2, R3, L1, and L2 are as defined in claim 9.

14. The use according to claim 13, characterized in that: The compound of formula (I) is selected from: Preferably, the compound of formula (I) is selected from:

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