Deuterated aminopyridine derivatives and pharmaceutical compositions containing the same

Abstract

The present invention relates to a deuterated aminopyridine derivative of formula I, its optical isomers or mixtures thereof, its crystal forms, its salts, its hydrates or solvates. The present invention also relates to a pharmaceutical composition comprising a pharmaceutically acceptable carrier and a deuterated aminopyridine derivative, its optical isomers or mixtures thereof, its crystal forms, its salts, its hydrates or solvates. The compound represented by general formula I of the present invention is a BTK inhibitor and is a prophylactic and / or therapeutic agent for BTK-related diseases, has low hepatotoxicity, and has good therapeutic effects on allergic diseases, autoimmune diseases, inflammatory diseases, thromboembolic diseases or cancers.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine, and particularly relates to deuterated aminopyridine derivatives and pharmaceutical compositions containing such compounds. Background Art

[0002] B cell signaling through the B cell receptor (BCR) can generate a wide range of biological output signals. Abnormal BCR-mediated signal transduction can result in dysregulated B cell activation and / or formation of pathogenic autoantibodies that lead to various autoimmune and / or inflammatory diseases. Mutations in BTK in humans result in X-linked agammaglobulinemia (XLA) (Conley et al., Annu. Rev. Immunol. 27:199-227, 2009). This disease is associated with impaired B cell maturation, reduced immunoglobulin production, impaired T cell-independent immune responses, and a marked attenuation of sustained calcium signaling upon BCR stimulation. Evidence for the role of BTK in allergic and / or autoimmune and / or inflammatory diseases has been established in BTK-deficient mouse models.

[0003] Due to the role of BTK in B cell activation, BTK inhibitors can be used as inhibitors of B cell-mediated pathogenic activities (such as autoantibody production). BTK is also expressed in osteoclasts, mast cells, and monocytes, and has been shown to be important for the function of these cells.

[0004] Therefore, inhibition of BTK activity can be used to treat allergic and / or autoimmune and / or inflammatory diseases, such as: rheumatoid arthritis, polyangiitis, idiopathic thrombocytopenic purpura (ITP), myasthenia gravis, allergic rhinitis, and asthma (Di Paolo et al. (2011) Nature Chem. Biol. 7(1):41-50; Liu et al. (2011) Jour. of Pharm. and Exper. Ther. 338(1):154-163).

[0005] In addition, the abnormal activation of BTK plays an important role in the pathogenesis of B-cell lymphoma, which means that inhibiting BTK is useful in the treatment of hematological malignancies (Davis et al., Nature 463:88-92, 2010). Since BTK plays a central role as a mediator in multiple signal transduction pathways, inhibiting BTK activity can be anti-inflammatory and / or anti-cancer, and is used for cancer and the treatment of B-cell lymphoma, leukemia, and other hematological malignancies (Mohamed et al., Immunol. Rev. 228:58-73, 2009; Pan, Drug News perspect 21: 357-362, 2008; Rokosz et al., Expert Opin. Ther. Targets 12:883-903, 2008; Uckun et al., Anti-cancer Agents Med. Chem. 7:624-632, 2007; Lou et al., J. Med. Chem. 55(10):4539-4550, 2012).

[0006] Moreover, considering the role of BTK in osteoclast function, inhibiting BTK activity can be used for the treatment of bone diseases, such as osteoporosis. Therefore, compounds with BTK inhibitory activity are useful for the treatment of diseases related to B cells and / or mast cells, such as allergic diseases, autoimmune diseases, inflammatory diseases, thromboembolic diseases, cancer, etc. (Uckun et al. (2007) Anticancer Agents in Medicinal Chemistry. 7(6):624-632).

[0007] Tolebrutinib (SAR442168, PRN2246) is an effective, selective, orally active, and blood-brain barrier-permeable Bruton's tyrosine kinase (BTK) inhibitor, a investigational brain-penetrant Bruton tyrosine kinase (BTK) inhibitor, and the first drug to complete a proof-of-concept study of BTK inhibitor for the treatment of multiple sclerosis (MS), which can produce the cerebrospinal fluid (CSF) concentration required to target microglia and B lymphocytes, and is undergoing clinical phase III trials for multiple sclerosis (MS) and myasthenia gravis (MG). However, drug-induced liver injury cases have occurred in patients with Tolebrutinib, and the US FDA has suspended some of its clinical trials. Improving the pharmacokinetic behavior of Tolebrutinib, such as the absorption and / or distribution and / or metabolism and / or excretion of Tolebrutinib, may be an effective way to reduce the toxicity of Tolebrutinib.

[0008] The human elimination half-life T1 / 2 of tolebrutinib is between 1.39 and 2.18 hours, and the elimination is rapid (Owens TD et al. Clin Transl Sci. 2022;15:442–450). This indicates that a relatively large dose is required to maintain a good therapeutic effect, but this inevitably brings a greater risk of toxicity. In addition, the short half-life and rapid elimination of tolebrutinib also to a certain extent suggest that the metabolism of tolebrutinib may be relatively fast, and these metabolites may also increase the risk of toxicity.

[0009] The suboptimal absorption, distribution, metabolism, and / or excretion (ADME) properties of some current drugs hinder their wider use or limit their application in specific indications. For example, due to the short elimination half-life and rapid clearance of drugs in the body, the commonly adopted solution is to administer the drug frequently or at a high dose to achieve a sufficiently high plasma drug level. However, this introduces a number of potential therapeutic problems, such as patient compliance with the dosing interval, and with higher-dose administration, side effects will be more severe, and the treatment cost is increased. Drugs with rapid metabolism may also expose patients to unwanted toxic or reactive metabolites.

[0010] Another ADME limitation affecting drugs is the formation of toxic or biologically reactive metabolites. Thus, some patients receiving the drug may experience toxicity, or the safe dose of such a drug may be limited such that the patient receives a suboptimal amount of treatment. In some cases, changing the dosing interval or formulation method may help reduce clinical adverse reactions, but the frequent formation of such unwanted metabolites is inherent in the metabolism of the compound.

[0011] A potentially attractive strategy for improving drug metabolic properties is deuterium modification. In this approach, one attempts to slow down the metabolism of the drug or reduce the formation of unwanted metabolites by replacing one or more hydrogen atoms with deuterium atoms. Deuterium is a safe, stable, non-radioactive isotope of hydrogen. Compared with hydrogen, deuterium forms stronger chemical bonds with carbon. In selected cases, the increased bond strength conferred by deuterium can positively affect the ADME properties of the drug, with the potential to improve efficacy, safety, and / or tolerability. At the same time, since the size and shape of deuterium are essentially equivalent to those of hydrogen, replacing hydrogen with deuterium is expected not to affect the biochemical potency and selectivity of the drug compared to the original chemical entity containing only hydrogen.

[0012] However, due to the complex metabolic processes of biological systems, the pharmacokinetic properties of drugs in vivo are affected by various factors and exhibit corresponding complexity. Compared with the corresponding non-deuterated drugs, the changes in the pharmacokinetic properties of deuterated drugs show great contingency and unpredictability. For some compounds, deuteration slows down their metabolic clearance in vivo and increases the half-life; for other compounds, deuteration does not cause metabolic changes; for still other compounds, deuteration accelerates metabolic clearance and shortens the half-life (Blake, MI et al, J Pharm Sci, 1975, 64: 367-91; Foster, AB, Adv Drug Res 1985, 14:1-40("Foster"); Kushner, DJ et al, Can J Physiol Pharmacol 1999, 79-88; Fisher, MB et al, Curr Opin Drug Discov Devel, 2006, 9:101-09("Fisher")).

[0013] Therefore, deuteration at certain sites of a compound may not only fail to increase the half-life but may even shorten it (Scott L. Harbeson, Roger D. Tung. Deuterium in Drug Discovery and Development, P405-406), deteriorating its pharmacokinetic properties; on the other hand, hydrogen at certain positions on the drug molecule is not easily deuterated due to reasons such as steric hindrance.

[0014] Even when deuterium atoms are incorporated into known metabolic sites, the effect of deuterium modification (deuterium modification) on drug metabolism is not predictable. Only by actually preparing and testing deuterated drugs can it be determined whether and how the metabolic rate will differ from that of the corresponding non-deuterated chemical entity. Many drugs have multiple sites where metabolism may occur. The positions (sites) that need to be deuterium-substituted and the degree of deuteration required to find an impact on metabolism, if any, will be different for each drug (Fukuto et al. J. Med. Chem. 1991, 34, 2871-76).

[0015] In addition, deuteration can lead to metabolic switching. The concept of metabolic switching suggests that when a drug is surrounded by phase I metabolic enzymes, it can transiently bind and rebind to the phase I metabolic enzymes in various conformations prior to a chemical reaction (such as an oxidation reaction). Therefore, metabolic switching can potentially result in different proportions of known metabolites as well as new metabolites. This new metabolic property can cause more or less toxicity and lead to a faster or slower drug clearance rate, thereby reducing or increasing the in vivo exposure of the drug. The changes caused by metabolic switching are unpredictable, and no sufficient a priori prediction has been achieved for any drug to date.

[0016] As mentioned above, the effect of deuterium modification (deuterium modification) on the metabolism of drugs is unpredictable. Tolebrutinib and its in vivo metabolites have the defect of hepatotoxicity risk, and the clinical trials of tolebrutinib have also shown hepatotoxicity, causing great clinical concern. Hepatotoxicity is not only related to the chemical structure but also closely related to the clinical administration dose.

[0017] Therefore, aiming at the deficiencies of the prior art, we design new compounds to increase the in vivo exposure of the new compounds, reduce the dosage and / or frequency of medication, and reduce hepatotoxicity; and / or through structural modification, reduce the hepatotoxicity of its prototype or metabolites, or reduce the generation of toxic metabolites, so as to achieve the purpose of reducing toxicity and enhancing efficacy. Summary of the Invention

[0018] The object of the present invention is to provide a class of novel compounds with BTK inhibitory activity and better pharmacodynamic properties and their uses.

[0019] In the first aspect of the present invention, there is provided a deuterated aminopyridine derivative represented by formula I, its optical isomers or mixtures thereof, its crystal forms, its salts, its hydrates or solvates.

[0020]

[0021] Wherein: R 1 、R 2 、R 3 、R 4 、R 5 、R 6 、R 7 、R 8 、R 9 、R 10 、R 11 、R 12 、R 13 、R 14 、R 15 、R 16 、R 17 、R 18or R 19 each independently selected from hydrogen (H) or deuterium (D), provided that R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , R 18 or R 19 at least one of which is deuterium.

[0022] In a preferred embodiment, the compound is a preferred compound selected from the group consisting of:

[0023]

[0024] In another preferred embodiment, the compound is a preferred compound selected from the group consisting of:

[0025]

[0026] In a second aspect of the present invention, there is provided a method for preparing a pharmaceutical composition, comprising the step of: mixing a pharmaceutically acceptable carrier with the compound, its optical isomers or mixtures thereof, its crystal forms, its salts, its hydrates or solvates as described in the first aspect of the present invention, thereby forming a pharmaceutical composition.

[0027] In a third aspect of the present invention, there is provided a pharmaceutical composition which contains a pharmaceutically acceptable carrier and the compound, its optical isomers or mixtures thereof, its crystal forms, its salts, its hydrates or solvates as described in the first aspect of the present invention.

[0028] In another preferred embodiment, the pharmaceutical composition is a capsule, tablet, injection, pill, powder or granule.

[0029] In a fourth aspect of the present invention, there is provided the use of the compound, its optical isomers or mixtures thereof, its crystal forms, its salts, its hydrates or solvates as described in the first aspect of the present invention for the preparation of a pharmaceutical composition for inhibiting BTK.

[0030] In another preferred embodiment, the pharmaceutical composition is used for preventing and / or treating diseases related to BTK.

[0031] In another preferred embodiment, the pharmaceutical composition is used for preventing and / or treating allergic disorders, autoimmune diseases, inflammatory diseases, thromboembolic diseases or cancers.

[0032] In another preferred embodiment, the pharmaceutical composition is used for treating autoimmune diseases, including multiple sclerosis (MS), myasthenia gravis (MG), chronic spontaneous urticaria, neuromyelitis optica, systemic lupus erythematosus (SLE), or rheumatoid arthritis (RA).

[0033] In another preferred embodiment, the pharmaceutical composition is used for treating cancers including (but not limited to): lymphoma, leukemia, non-small cell lung cancer, uterine cancer, rectal cancer, brain cancer, head cancer, cervical cancer, bladder cancer, prostate cancer, breast cancer, kidney cancer, liver cancer, gastric cancer, or pancreatic cancer.

[0034] In a fifth aspect of the present invention, there is provided a treatment method, which comprises the step of administering to a subject in need of treatment the compound described in the first aspect of the present invention, its optical isomers or mixtures thereof, its crystal forms, its salts, its hydrates or solvates, or administering the pharmaceutical composition described in the third aspect of the present invention, thereby inhibiting BTK.

[0035] As used herein, "deuterated" means that one or more hydrogens in a compound or group are replaced by deuterium. Deuteration can be mono-substitution, di-substitution, multi-substitution or full substitution.

[0036] In another preferred embodiment, the deuterium isotope content of deuterium at the deuterium substitution position is greater than the natural deuterium isotope content (0.015%), more preferably greater than 50%, more preferably greater than 85%, more preferably greater than 95%, more preferably greater than 99%, more preferably greater than 99.5%.

[0037] In another preferred embodiment, the compound of formula I contains at least 1 or 3 deuterium atoms, more preferably 5 or 8 deuterium atoms.

[0038] As used herein, the term "compound of the present invention" refers to the compound shown in formula I. This term also includes the optical isomers or mixtures thereof, crystal forms, salts, hydrates or solvates of the compound of formula I.

[0039] As used herein, the term "pharmaceutically acceptable salt" refers to a salt formed by the compound of the present invention with an acid or a base that is suitable for use as a drug. Pharmaceutically acceptable salts include inorganic salts and organic salts. A preferred class of salts are the salts formed by the compound of the present invention with acids. Acids suitable for forming salts include but are not limited to: inorganic acids such as hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid, phosphoric acid, etc., organic acids such as formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, picric acid, methanesulfonic acid, benzenesulfonic acid, benzene sulfonic acid, etc.; and acidic amino acids such as aspartic acid and glutamic acid.

[0040] The positive and progressive effects of the present invention are as follows:

[0041] (1) The compounds of the present invention have good selective BTK inhibitory effects and can be effectively used for diseases related to BTK.

[0042] (2) The compounds of the present invention have good selective inhibitory effects on B cell activation and are effective as B cell activation inhibitors.

[0043] (3) The deuterated aminopyridine derivatives of the present invention have low hepatotoxicity, good pharmacokinetic properties, reduced dosage and / or reduced toxic and side effects, and better drug-likeness.

[0044] Specific implementation methods

[0045] The present invention will be further described below in conjunction with specific embodiments. It should be understood that these embodiments are only used to illustrate the present invention and not to limit the scope of the present invention. The experimental methods without specific conditions noted in the following embodiments are generally carried out under conventional conditions or according to the conditions recommended by the manufacturers.

[0046] The preparation methods of the non-deuterated pyrimidine derivatives and their physiologically compatible salts used in the present invention are known. The corresponding deuterated pyrimidine derivatives can be synthesized by the same route using the corresponding deuterated starting compounds as raw materials.

[0047] The preparation methods of the non-deuterated pyrimidine derivatives and their physiologically compatible salts used in the present invention are known. The corresponding deuterated pyrimidine derivatives can be synthesized by the same route using the corresponding deuterated starting compounds as raw materials.

[0048] Example 1: Synthesis of Compound T109

[0049] Synthesis route

[0050]

[0051] Step 1: Synthesis of Compound T003

[0052] To N,N-dimethylformamide (50 ml), 2,4-dichloro-3-nitropyridine (41.5 mmol), tert-butyl (R)-3-aminopiperidine-1-carboxylate (41.4 mmol) and TEA (62.2 mmol) were added. The resulting reaction mixture was stirred overnight at 25 °C. The reaction mixture was diluted with water, extracted with ethyl acetate, and the organic layers were combined, washed with saturated sodium chloride, dried over anhydrous sodium sulfate and concentrated. Purification by silica gel column chromatography, eluent ethyl acetate / petroleum ether (1:1), removal of the solvent gave the compound T003 as a yellow oil.

[0053] Step 2: Synthesis of compound T005

[0054]

[0055] To isopropanol (100 ml), compound T003 (22.4 mmol), bis[(4-methoxyphenyl)methyl]amine (compound T004) (22.4 mmol) and TEA (29.5 mmol) were added. The resulting reaction mixture was stirred overnight at 95 °C, cooled and concentrated in vacuo to give the compound T005 as a yellow oil.

[0056] Step 3: Synthesis of compound T006

[0057]

[0058] To a solution of AcOH / MeOH (1:1, 100 mL), compound T005 (17.3 mmol) and Fe (173.1 mmol) were added. The reaction mixture was stirred overnight at 25 °C and then concentrated in vacuo. The pH of the residual solution was adjusted to 8.0 - 9.0 with sodium bicarbonate. The resulting solution was extracted with dichloromethane, and the organic layer was washed with sodium bicarbonate, dried over anhydrous sodium sulfate and concentrated in vacuo to give the compound T006 as a yellow oil.

[0059] Step 4: Synthesis of compound T007

[0060]

[0061] To CH3CN (100 ml), compound T006 (20.2 mmol) and CDI (30.1 mmol) were added. The reaction mixture was stirred overnight at 80 °C. The reaction mixture was cooled and concentrated. Purification by silica gel column chromatography, eluent ethyl acetate / petroleum ether (1:5), removal of the solvent gave the compound T007.

[0062] Step 5: Synthesis of compound T009

[0063]

[0064] To dichloromethane (100 ml), add compound T007 (17.5 mmol), pentadeuterated (4-phenoxyphenyl)boronic acid (compound T008) (35.1 mmol), TEMPO (19.5 mmol), TEA (69.5 mmol), and Cu(OAc)2 (8.9 mmol). Stir the reaction mixture at 25 °C overnight under ambient pressure in an oxygen atmosphere. Add pentadeuterated (4-phenoxyphenyl)boronic acid (compound T008) (35.1 mmol) again, and allow the reaction mixture to react at 25 °C overnight. Purify by silica gel column chromatography, with the eluent being ethyl acetate / petroleum ether (1:3). Remove the solvent to obtain compound T009.

[0065] Step 6: Synthesis of compound T010

[0066]

[0067] To dichloromethane (80 ml), add compound T009 (6.1 mmol) and trifluoroacetic acid (80 ml). Stir the resulting reaction mixture at 50 °C for 5 hours, concentrate in vacuo, adjust the pH of the residual solution to 9 with sodium bicarbonate, extract the resulting solution with dichloromethane, combine the organic layers, and dry over anhydrous sodium sulfate. Purify by silica gel column chromatography, with the eluent being dichloromethane / methanol (30:1). Remove the solvent to obtain compound T010.

[0068] Step 7: Synthesis of compound T109

[0069]

[0070] To DCM-CH3OH (1:1, 6 ml), add compound T010 (150 mg, 0.37 mmol, 1.00 equivalent), TEA (113 mg, 1.12 mmol, 3.00 equivalents). Then, dropwise add trideuterated prop-2-enoyl chloride (compound T011) (40.1 mg, 0.44 mmol, 1.20 equivalents) with stirring at 0 °C over 5 minutes. Stir the resulting solution at 0 °C for 2 hours and concentrate in vacuo. Purify the residue by silica gel column chromatography, with the eluent being dichloromethane / methanol (30:1). Remove the solvent to obtain the crude product. The crude product is purified by preparative C 18 chromatography column, with the mobile phase being 0.05% TFA and ACN, and gradient elution. Compound T109 is obtained.

[0071] The 1H NMR spectrum of compound T109 is as follows: 1 H-NMR (DMSO- d6) δ 7.8 (1H), 7.4 - 7.5 (2H), 7.1 - 7.3 (2H), 7.0 (1H), 5.0 (2H), 4.5 (1H), 4.2 (2H), 3.8 (0.5H), 3.2 (1H), 2.7 (0.5H), 2.4 (1H), 1.9 (2H), 1.6 (1H).

[0072] Example 2: Synthesis of Compound T108

[0073] Synthesis of compound T108: In the synthesis of “Example 1: Synthesis of compound T109”, change compound T008 in step 5 to compound T201. The remaining steps are the same as the operation in “Example 1: Synthesis of compound T109” to obtain compound T108. The nuclear magnetic resonance hydrogen spectrum of compound T108 is as follows: 1 H-NMR (DMSO- d 6) δ 7.8 (1H), 7.4 - 7.5 (2H), 7.1 - 7.3 (4H), 7.0 (1H), 5.0 (2H), 4.5 (1H), 4.2 (2H), 3.8 (0.5H), 3.2 (1H), 2.7 (0.5H), 2.4 (1H), 1.9 (2H), 1.6 (1H).

[0074]

[0075] Example 3: Synthesis of Compound T107

[0076] Synthesis of compound T107: In the synthesis of “Example 1: Synthesis of compound T109”, change compound T008 in step 5 to compound T202. The remaining steps are the same as the operation in “Example 1: Synthesis of compound T109” to obtain compound T107. The nuclear magnetic resonance hydrogen spectrum of compound T107 is as follows: 1 H-NMR (DMSO- d 6) δ 7.8 (1H), 7.4 - 7.5 (4H), 7.1 - 7.3 (2H), 7.0 (1H), 5.0 (2H), 4.5 (1H), 4.2 (2H), 3.8 (0.5H), 3.2 (1H), 2.7 (0.5H), 2.4 (1H), 1.9 (2H), 1.6 (1H).

[0077]

[0078] Example 4: Synthesis of Compound T106

[0079] Synthesis of compound T106: In the synthesis of “Example 1: Synthesis of compound T109”, change compound T008 in step 5 to compound T203. The remaining steps are the same as those in the operation of “Example 1: Synthesis of compound T109” to obtain compound T106. The nuclear magnetic resonance hydrogen spectrum of compound T106 is as follows: 1 H-NMR (DMSO- d 6) δ 7.8(1H), 7.4 - 7.5(4H), 7.1 - 7.3(4H), 7.0(1H), 5.0(2H), 4.5(1H), 4.2(2H), 3.8(0.5H), 3.2(1H), 2.7(0.5H), 2.4(1H), 1.9(2H), 1.6(1H).

[0080]

[0081] Example 5: Synthesis of Compound T105

[0082] Synthesis of compound T105: In the synthesis of “Example 1: Synthesis of compound T109”, change compound T008 in step 5 to compound T204. The remaining steps are the same as those in the operation of “Example 1: Synthesis of compound T109” to obtain compound T105. The nuclear magnetic resonance hydrogen spectrum of compound T105 is as follows: 1 H-NMR (DMSO- d 6) δ 7.8(1H), 7.4 - 7.5(4H), 7.1 - 7.3(5H), 7.0(1H), 5.0(2H), 4.5(1H), 4.2(2H), 3.8(0.5H), 3.2(1H), 2.7(0.5H), 2.4(1H), 1.9(2H), 1.6(1H).

[0083]

[0084] Example 6: Synthesis of Compound T104

[0085] Synthesis of compound T104: In the synthesis of “Example 1: Synthesis of compound T109”, change compound T011 in step 7 to compound T205. The remaining steps are the same as those in the operation of “Example 1: Synthesis of compound T109” to obtain compound T104. The nuclear magnetic resonance hydrogen spectrum of compound T104 is as follows: 1 H-NMR (DMSO- d6) δ 7.8 (1H), 7.4 - 7.5 (2H), 7.1 - 7.3 (2H), 7.0 (1H), 6.8 (1H), 6.1 (1H), 5.7 (1H), 5.0 (2H), 4.5 (1H), 4.2 (2H), 3.8 (0.5H), 3.2 (1H), 2.7 (0.5H), 2.4 (1H), 1.9 (2H), 1.6 (1H).

[0086]

[0087] Example 7: Synthesis of Compound T103

[0088] Synthesis of compound T103: In the synthesis of “Example 6: Synthesis of compound T104”, change compound T008 in step 5 to compound T201. The remaining steps are the same as those in the operation of “Example 6: Synthesis of compound T104” to obtain compound T103. The 1H-NMR spectrum of compound T103 is as follows: 1 1H-NMR (DMSO- d 6) δ 7.8 (1H), 7.4 - 7.5 (2H), 7.1 - 7.3 (4H), 7.0 (1H), 6.8 (1H), 6.1 (1H), 5.7 (1H), 5.0 (2H), 4.5 (1H), 4.2 (2H), 3.8 (0.5H), 3.2 (1H), 2.7 (0.5H), 2.4 (1H), 1.9 (2H), 1.6 (1H).

[0089]

[0090] Example 8: Synthesis of Compound T102

[0091] Synthesis of compound T102: In the synthesis of “Example 6: Synthesis of compound T104”, change compound T008 in step 5 to compound T202. The remaining steps are the same as those in the operation of “Example 6: Synthesis of compound T104” to obtain compound T102. The 1H-NMR spectrum of compound T102 is as follows: 1 1H-NMR (DMSO- d 6) δ 7.8 (1H), 7.4 - 7.5 (4H), 7.1 - 7.3 (2H), 7.0 (1H), 6.8 (1H), 6.1 (1H), 5.7 (1H), 5.0 (2H), 4.5 (1H), 4.2 (2H), 3.8 (0.5H), 3.2 (1H), 2.7 (0.5H), 2.4 (1H), 1.9 (2H), 1.6 (1H).

[0092] Example 9: Synthesis of Compound T101

[0093] Synthesis of compound T101: In the synthesis of “Example 6: Synthesis of compound T104”, change compound T008 in step 5 to compound T203. The remaining steps are the same as the operation in “Example 6: Synthesis of compound T104” to obtain compound T101. The proton nuclear magnetic resonance spectrum of compound T101 is as follows: 1 H-NMR (DMSO- d 6) δ 7.8 (1H), 7.4 - 7.5(4H), 7.1 - 7.3(4H), 7.0(1H), 6.8 (1H), 6.1 (1H), 5.7(1H), 5.0 (2H), 4.5 (1H), 4.2(2H), 3.8(0.5H), 3.2(1H), 2.7 (0.5H), 2.4(1H), 1.9 (2H), 1.6(1H).

[0094]

[0095] Example 10: Synthesis of Compound T113

[0096] Synthesis of compound T113: In the synthesis of “Example 6: Synthesis of compound T104”, change compound T008 in step 5 to compound T206. The remaining steps are the same as the operation in “Example 6: Synthesis of compound T104” to obtain compound T113. The proton nuclear magnetic resonance spectrum of compound T113 is as follows: 1 H-NMR (DMSO- d 6) δ 7.8 (1H), 7.4 - 7.5(2H), 7.1 - 7.3(3H), 7.0(1H), 6.8 (1H), 6.1 (1H), 5.7(1H), 5.0 (2H), 4.5 (1H), 4.2(2H), 3.8(0.5H), 3.2(1H), 2.7 (0.5H), 2.4(1H), 1.9 (2H), 1.6(1H).

[0097]

[0098] Example 11: Synthesis of Compound T114

[0099] Synthesis of compound T114: In the synthesis of “Example 6: Synthesis of compound T104”, change compound T008 in step 5 to compound T207. The remaining steps are the same as the operation in “Example 6: Synthesis of compound T104” to obtain compound T114. The proton nuclear magnetic resonance spectrum of compound T114 is as follows: 1 H-NMR (DMSO- d6) δ 7.8 (1H), 7.4 - 7.5(4H), 7.1 - 7.3(3H), 7.0(1H), 6.8 (1H), 6.1 (1H), 5.7(1H), 5.0 (2H), 4.5 (1H), 4.2(2H), 3.8(0.5H), 3.2(1H), 2.7 (0.5H), 2.4(1H), 1.9 (2H), 1.6(1H).

[0100]

[0101] Example 12: Synthesis of Compound T115

[0102] For the synthesis of compound T115, change compound T008 in step 5 of "Example 6: Synthesis of compound T104" to compound T208. The remaining steps are the same as those in "Example 6: Synthesis of compound T104". Compound T115 is obtained. The 1H-NMR spectrum of compound T115 is as follows: 1 H-NMR (DMSO- d 6) δ 7.8 (1H), 7.4 - 7.5(2H), 7.1 - 7.3(5H), 7.0(1H), 6.8 (1H), 6.1 (1H), 5.7(1H), 5.0 (2H), 4.5 (1H), 4.2(2H), 3.8(0.5H), 3.2(1H), 2.7 (0.5H), 2.4(1H), 1.9 (2H), 1.6(1H).

[0103]

[0104] Example 13: Synthesis of Compound T112

[0105] For the synthesis of compound T112, change compound T008 in step 5 of "Example 1: Synthesis of compound T109" to compound T208. The remaining steps are the same as those in "Example 1: Synthesis of compound T109". Compound T112 is obtained. The 1H-NMR spectrum of compound T112 is as follows: 1 H-NMR (DMSO- d 6) δ 7.8 (1H), 7.4 - 7.5(2H), 7.1 - 7.3(5H), 7.0(1H), 5.0 (2H), 4.5 (1H), 4.2(2H), 3.8(0.5H), 3.2(1H), 2.7 (0.5H), 2.4(1H), 1.9 (2H), 1.6(1H).

[0106]

[0107] Example 14: Synthesis of Compound T111

[0108] Synthesis of compound T111: In the synthesis of “Example 1: Synthesis of compound T109”, change compound T008 in step 5 to compound T207. The remaining steps are the same as those in “Example 1: Synthesis of compound T109” to obtain compound T111. The proton nuclear magnetic resonance spectrum of compound T111 is as follows: 1 H-NMR (DMSO- d 6) δ 7.8 (1H), 7.4 - 7.5(4H), 7.1 - 7.3(3H), 7.0(1H), 5.0 (2H), 4.5 (1H), 4.2(2H), 3.8(0.5H), 3.2(1H), 2.7 (0.5H), 2.4(1H), 1.9 (2H), 1.6(1H).

[0109]

[0110] Example 15: Synthesis of Compound T110

[0111] Synthesis of compound T110: In the synthesis of “Example 1: Synthesis of compound T109”, change compound T008 in step 5 to compound T206. The remaining steps are the same as those in “Example 1: Synthesis of compound T109” to obtain compound T110. The proton nuclear magnetic resonance spectrum of compound T110 is as follows: 1 H-NMR (DMSO- d 6) δ 7.8 (1H), 7.4 - 7.5(2H), 7.1 - 7.3(3H), 7.0(1H), 5.0 (2H), 4.5 (1H), 4.2(2H), 3.8(0.5H), 3.2(1H), 2.7 (0.5H), 2.4(1H), 1.9 (2H), 1.6(1H).

[0112]

[0113] Example 16: Synthesis of Compound T008

[0114]

[0115] Step 8: Synthesis of compound T032

[0116] Take phenol-d5 (Compound T030) (9.4 g), dissolve it in anhydrous tetrahydrofuran (100 ml), stir, and slowly add sodium hydride (9.6 g) in batches. Then add 1-bromo-4-iodobenzene (Compound T031) (31.1 g) in batches. React at room temperature for 15 hr. Filter the reactant, rotary evaporate the filtrate, and dissolve it in dichloromethane. Column chromatography is carried out, and elute with petroleum ether:ethyl acetate (1:5) to obtain Compound T032.

[0117] Step 9: Synthesis of Compound T008

[0118] Dissolve Compound T032 (4.5 g) in dry THF (100 ml), react at -78 °C for 30 min under N2 protection, slowly add n-butyllithium (1.7 g), keep reacting at -78 °C for 3 h after the addition, then slowly add triisopropyl borate (3.8 g), keep reacting at -78 °C for 2 h after the addition, slowly warm to room temperature, and react for about 15 hr. After monitoring the reaction by TLC and the reaction is completed, slowly quench the reaction solution with water, extract and concentrate to obtain Compound T008.

[0119] Example 17: Synthesis of Compound T202

[0120] The synthesis of Compound T202 is carried out according to "Example 16: Synthesis of Compound T008", only in Step 8, Compound T030 is changed to Compound T033, and the remaining steps are the same as the operation in "Example 16: Synthesis of Compound T008" to obtain Compound T202.

[0121]

[0122] Example 18: Synthesis of Compound T201

[0123] The synthesis of Compound T201 is carried out according to "Example 16: Synthesis of Compound T008", only in Step 8, Compound T030 is changed to Compound T034, and the remaining steps are the same as the operation in "Example 16: Synthesis of Compound T008" to obtain Compound T201.

[0124]

[0125] Example 19: Synthesis of Compound T203

[0126] The synthesis of Compound T203 is carried out according to "Example 16: Synthesis of Compound T008", only in Step 8, Compound T030 is changed to Compound T035, and the remaining steps are the same as the operation in "Example 16: Synthesis of Compound T008" to obtain Compound T203.

[0127]

[0128] Example 20: Synthesis of Compound T206

[0129] The synthesis of compound T206 was carried out according to "Example 16: Synthesis of compound T008", except that in step 8, compound T030 was changed to compound T036, and the remaining steps were the same as those in "Example 16: Synthesis of compound T008" to obtain compound T206.

[0130]

[0131] Example 21: Synthesis of Compound T207

[0132] The synthesis of compound T207 was carried out according to "Example 16: Synthesis of compound T008", except that in step 8, compound T030 was changed to compound T037, and the remaining steps were the same as those in "Example 16: Synthesis of compound T008" to obtain compound T207.

[0133]

[0134] Example 22: Synthesis of Compound T208

[0135] The synthesis of compound T208 was carried out according to "Example 16: Synthesis of compound T008", except that in step 8, compound T030 was changed to compound T038, and the remaining steps were the same as those in "Example 16: Synthesis of compound T008" to obtain compound T208.

[0136]

[0137] Example 23: Pharmacokinetic Evaluation in Rats

[0138] Forty-eight male Sprague-Dawley rats, 7 - 8 weeks old and weighing about 210 g, were divided into 8 groups (Tolebrutinib group, compound T101 group, compound T104 group, compound T105 group, compound T106 group, compound T107 group, compound T109 group and compound T112 group), with 6 rats in each group. Tolebrutinib, compound T101, compound T104, compound T105, compound T106, compound T107, compound T109 and compound T112 were administered by single intragastric gavage at a dose of 6 mg / kg according to the grouping, and their pharmacokinetic differences were compared.

[0139] The rats were fed with standard feed and fasted 12 hours before dosing. The dosing solution was prepared with 0.5% sodium carboxymethylcellulose (CMC-Na). Blood was collected from the orbital venous plexus at the time points of 0.25 hour, 0.5 hour, 1 hour, 2 hours, 4 hours, 6 hours, 8 hours, 10 hours and 24 hours after dosing.

[0140] After blood sampling, place it in a centrifuge tube coated with heparin sodium solution, gently invert the blood collection tube at least 5 times immediately, ensure thorough mixing, and then place it on ice. Centrifuge the blood sample at 5000 revolutions per minute for 5 minutes at 4 °C to separate the plasma from the red blood cells. Use a pipette to aspirate 100 μL of plasma into a clean plastic centrifuge tube, and label the sample number and blood collection time point. The plasma is stored in a -80 °C refrigerator before LC-MS / MS analysis.

[0141] As can be seen from the test results, compared with Tolebrutinib, for compound T101, the elimination half-life T 1 / 2 and / or the area under the curve AUC and / or the maximum plasma concentration C max increased by more than 40%; compared with Tolebrutinib, for compound T104, the elimination half-life T 1 / 2 and / or the area under the curve AUC and / or the maximum plasma concentration C max increased by more than 40%; compared with Tolebrutinib, for compound T105, the elimination half-life T 1 / 2 and / or the area under the curve AUC and / or the maximum plasma concentration C max increased by more than 50%; compared with Tolebrutinib, for compound T106, the elimination half-life T 1 / 2 and / or the area under the curve AUC and / or the maximum plasma concentration C max increased by more than 50%; compared with Tolebrutinib, for compound T107, the elimination half-life T 1 / 2 and / or the area under the curve AUC and / or the maximum plasma concentration C max increased by more than 40%; compared with Tolebrutinib, for compound T109, the elimination half-life T 1 / 2 and / or the area under the curve AUC and / or the maximum plasma concentration C max increased by more than 50%; compared with Tolebrutinib, for compound T112, the elimination half-life T 1 / 2 and / or the area under the curve AUC and / or the maximum plasma concentration C max increased by more than 40%.

[0142] As can be seen from these results, the compounds T101, T104, T105, T106, T107, T109, and T112 of the present invention have better pharmacokinetic properties in animals, indicating better pharmacodynamic and therapeutic effects.

[0143] Example 24: Determination of BTK Inhibitory Activity

[0144] The ADP-Glo™ kit was used to determine the effect of the compounds of the present invention on the activity of BTK. The experimental method is as follows:

[0145] ADP is the product of the kinase reaction, and the kinase activity can usually be detected by measuring the amount of ADP generated. The ADP-Glo™ kit developed by Promega Corporation measures the in vitro activity of BTK by detecting the level of ADP generated in the kinase reaction. In the kinase assay, the kinase consumes ATP to phosphorylate the substrate, generating ADP at the same time. Then, the ADP-Glo reagent is added to terminate the kinase reaction and completely consume the remaining ATP. Next, the kinase detection reagent is added to convert the generated ADP into new ATP. The luciferase in the detection reagent can catalyze luciferin in the presence of ATP and O2 to produce a light signal, thus converting the chemical signal into a light signal. The intensity of the light signal is positively correlated with the amount of ADP generated in the kinase reaction, enabling the quantitative detection of the activity of the kinase BTK.

[0146] All detection experiments were carried out at a constant room temperature of 23 °C. A Corning 3674 white 384-well detection plate, the kinase BTK (Invitrogen), the kinase substrate being the polypeptide (4:1 Glu, Tyr) (Signal Chem) and ATP (Sigma), and an EnVision microplate reader (Perkin Elmer) were used to read the light signal. The detection buffer included 40 mM Tris-HCl (pH 7.5), 10 mM MgCl2 (Sigma), 2 mM MnCl2 (Sigma), 0.05 mM DTT (Sigma), and 0.01% BSA (Sigma); the kinase BTK was prepared as a kinase reaction solution with a concentration of 1.3 ng / μL using the detection buffer; the substrate reaction solution included 0.25 mg / mL polypeptide substrate and 60 μM ATP.

[0147] The compounds of the present invention were diluted with DMSO to a solution of 0.5 mM, and then serially diluted three-fold with DMSO to a minimum concentration of 0.025 μM. 50 nL of the compound solution with a series of concentrations and 2.5 μL of the kinase reaction solution were first added to the 384-well plate using an Echo555. After mixing evenly, the mixture was incubated at room temperature in the dark for 30 minutes; then 2.5 μL of the substrate reaction solution was added, and the total reaction volume was 5.05 μL. The reaction mixture was reacted at room temperature in the dark for 60 minutes; then 5 μL of the ADP-Glo™ reagent was added to terminate the reaction. After mixing evenly, the mixture was placed at room temperature for 40 minutes; finally, 10 μL of the kinase detection reagent was added, and the mixture was placed at room temperature in the dark for 30 minutes, and then the value was read on the Envision.

[0148] The inhibition percentage is calculated according to the following formula:

[0149] Inhibition % = [1 - (RLU 化合物 - RLU min ) / (RLU max - RLU min )] × 100

[0150] Where RLU 化合物 is the reading at a given concentration of the compound of the present invention, RLU min is the reading without adding the kinase BTK, and RLU max is the reading without adding the compound of the present invention. The IC 50 value of the compound is calculated by using the XLfit program in Excel.

[0151] Table 1: IC 50 values of the compounds of the present invention

[0152] Table 1: IC 50 values of the compounds of the present invention

[0153]

[0154] From these results, it can be seen that the compounds of the present invention have a significant inhibitory effect on BTK.

[0155] Example 25: Comparative Study of Mouse Hepatotoxicity

[0156] (1) Experimental animals

[0157] Thirty-two adult male ICR mice, weighing (25 ± 2 g), were selected. All mice were allowed free access to food, water and maintenance feed, and were kept under a day-night cycle at a temperature of 25 ± 2 °C and a relative humidity of 50 ± 10%.

[0158] (2) Animal grouping and drug administration

[0159] Thirty-two male ICR mice were divided into four groups of 8 mice each, namely a normal control group, a model group, a model + example compound group, and a model + Tolebrutinib group. In the model + example compound group, the example compound was administered by gavage once a day at a dose of (50 mg / kg); in the model + Tolebrutinib group, Tolebrutinib was administered by gavage once a day at a dose of (50 mg / kg), for 8 - 16 weeks respectively. The normal control group and the model group were respectively gavaged with an equal volume of pure water. After the last administration, food was withheld. One hour later, mice in the model group, the model + example compound group, and the model + Tolebrutinib group were intraperitoneally injected once with an APAP saline solution at a dose of 250 mg / kg. Twenty-four hours after modeling, blood was collected from the eyes of mice in each group in turn. The serum was separated by centrifugation at 3000 r / min for 10 min and stored at 4°C for later use; the liver and spleen were quickly dissected. After rinsing with normal saline at 4°C and blotting dry with filter paper, they were weighed. A part of the liver was fixed in 10% formaldehyde solution for sectioning, and the remaining liver was stored in a -80°C low-temperature refrigerator.

[0160] (3) Determination of biochemical indexes in the liver:

[0161] A part of the liver was weighed, and 9 times the volume of ice-cold normal saline was added. A 10% liver tissue homogenate was prepared using a tissue homogenizer, and the supernatant was obtained by centrifugation. The OD value was measured at 450 nm by spotting on a plate according to the kit method, and the content of MDA and the activity of GSH in the liver were calculated according to the formula.

[0162] (4) Data processing

[0163] Experimental data were expressed as mean ± standard deviation (±s), and analyzed using SPSS 22.0 statistical software. One-way ANOVA was used to compare the differences between groups. P < 0.05 was considered a significant difference.

[0164] (5) Effect of the example compound of the present application on lipid peroxidation of liver tissue in APAP-induced liver injury mice

[0165] Compared with the normal control group, the content of MDA in the liver tissue homogenate of the model group mice increased significantly, and the level of GSH decreased significantly (P<0.05), resulting in the accumulation of lipid peroxidation products in the mice and the reduction of the antioxidant metabolism level; compared with the model group, there were no significant changes in the MDA content and GSH level in the model + example compound group (P>0.05); compared with the model group, the MDA content in the model + Tolebrutinib group increased significantly (P<0.05), and the GSH level decreased significantly (P<0.05), indicating that the example compound (50 mg / kg) of the present application had no obvious effect on the lipid peroxidation induced by APAP, while Tolebrutinib (50 mg / kg) had an effect on the lipid peroxidation induced by APAP, suggesting that the liver toxicity of the example compound of the present application in mice was less than that of Tolebrutinib. The results are shown in Table 2.

[0166] Table 2. Effects on lipid peroxidation in liver tissue of APAP-induced liver injury mice

[0167]

[0168] Annotation (grade): A+ is 2.6 - 3.7; A- is 0.9 - 2.4; A++ is 3.9 - 4.7; B+ is 29 - 38; B- is 40 - 57; B++ is 13 - 27.

[0169] Conclusion: The example compound (50 mg / kg) of the present application has no obvious effect on the lipid peroxidation induced by APAP, while Tolebrutinib (50 mg / kg) has an effect on the lipid peroxidation induced by APAP, suggesting that the liver toxicity of the example compound of the present application in mice is less than that of Tolebrutinib.

[0170] Finally, it is necessary to note that the above detailed description of the specific embodiments of the present invention is only an example, and the present invention is not limited to the specific embodiments described above. For those skilled in the art, any equivalent modifications and substitutions made to the present invention are also within the scope of the present invention. Therefore, all equivalent transformations and modifications made without departing from the spirit and scope of the present invention should be covered within the scope of the present invention.

Claims

1. A compound or a pharmaceutically acceptable salt thereof, characterized in that, The compound is one of the compounds shown below: 。 2. A pharmaceutical composition, characterized in that Comprising the compound according to claim 1 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or excipient.

3. Use of the compound according to claim 1 or a pharmaceutically acceptable salt thereof in the preparation of a BTK inhibitor.

4. Use of the compound according to claim 1 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating and / or preventing a disease associated with BTK.

5. The application according to claim 4, wherein The disease associated with BTK is an allergic disorder, an autoimmune disease, an inflammatory disease, a thromboembolic disease or cancer.

6. The application according to claim 5, wherein The autoimmune disease is selected from multiple sclerosis, systemic lupus erythematosus, chronic spontaneous urticaria, neuromyelitis optica or rheumatoid arthritis.

7. The application according to claim 5, characterized in that, The cancer is selected from lymphoma, leukemia, non-small cell lung cancer, uterine cancer, rectal cancer, brain cancer, head cancer, cervical cancer, bladder cancer, prostate cancer, breast cancer, kidney cancer, liver cancer, gastric cancer or pancreatic cancer.

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