Pyridineamides as BTK inhibitors

Abstract

The present invention relates to pyridine amide compounds as Bruton's tyrosine kinase (BTK) inhibitors. Specifically, the present invention discloses deuterated compounds of 2-(4-phenoxyphenyl)-6-[1-(prop-2-enoyl)piperidin-4-yl]pyridine-3-carboxamide and / or its tautomers, pharmaceutical compositions of their crystal forms, salts, hydrates or solvates, and their use as preventive and / or therapeutic agents for BTK-related diseases.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine, and specifically relates to a pyridine amide compound used as a BTK inhibitor. Background Art

[0002] B cell signal transduction via the B cell receptor (BCR) can produce a wide range of biological output signals, and abnormal BCR-mediated signal transduction can cause dysregulated B cell activation and / or the formation of pathogenic autoantibodies that lead to a variety of autoimmune diseases and / or inflammatory diseases. Mutations in BTK in humans lead to 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 immune responses independent of T cells, and a significant reduction in sustained calcium signals during BCR stimulation.

[0003] Inhibition of BTK activity can be used to treat allergic diseases and / or autoimmune diseases 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).

[0004] In addition, abnormal activation of BTK plays an important role in the pathogenesis of B-cell lymphoma, which means that inhibition of BTK is very 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 can be 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).

[0005] 2-(4-phenoxyphenyl)-6-[1-(prop-2-enoyl)piperidin-4-yl]pyridine-3-carboxamide (Orelabrutinib, ICP-022) is a Bruton's tyrosine kinase (BTK) inhibitor used to treat lymphoma, leukemia and autoimmune diseases. Its elimination half-life in rats is 8.25 hours (Ya-nan Liu et al. (2022), Frontiers in Pharmacology.10.3389 / fphar.2022.991281), and the effects of Orelabrutinib drug metabolism and kinetics on efficacy and / or toxicity are still not very clear, and there is the possibility of improvement.

[0006] The poor pharmacokinetic and pharmacodynamic properties of some current drugs, such as absorption, distribution, metabolism and / or excretion, prevent 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 used solution is to give drugs frequently or in high doses to obtain sufficiently high plasma levels of drugs. However, this introduces a large number of potential treatment problems, such as patient compliance with medication intervals, and higher doses, more severe side effects, and increased treatment costs.

[0007] People try to slow down the metabolism of drugs by deuterium modification (modification), or by replacing one or more hydrogen atoms with deuterium atoms to reduce the formation of undesirable metabolites, etc., so as to improve the drug metabolism and kinetic properties of drugs. Deuterium is a safe, stable, non-radioactive isotope of hydrogen. Compared with hydrogen, deuterium forms a stronger chemical bond with carbon. In selected cases, the increased bond strength given by deuterium can improve the drug metabolism and kinetic properties of drugs, with the possibility of improving drug efficacy, safety, and / or tolerance. At the same time, because the size and shape of deuterium are substantially equivalent to hydrogen, compared with the original chemical entity containing only hydrogen, it is expected that replacing hydrogen with deuterium will not affect the biochemical efficacy and selectivity of drugs.

[0008] However, due to the complex metabolic process of biological systems, the pharmacokinetic properties of drugs in vivo are affected by many factors and also show 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 and half-life in the body; for other compounds, deuteration does not cause metabolic changes; for other other compounds, deuteration speeds up metabolic clearance and shortens 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 Dev , 2006, 9: 101-09 ("Fisher")).

[0009] Therefore, deuterium substitution at certain sites of the compound not only fails to increase the half-life, but may 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 substituted by deuterium due to steric hindrance and other reasons.

[0010] Even when deuterium atoms are incorporated into known metabolic sites, the effect of deuterium modification (deuterium modification) on the metabolism of drugs is not predictable. Only by actually preparing and testing deuterated drugs can it be determined whether and how the rate of metabolism will be different from the corresponding chemical entity of non-deuterated. Many drugs have multiple sites that may be metabolized. The position (site) where deuterium substitution is required and the degree of deuteration required to find the effect on metabolism, if any, will be different for each drug (Fukuto et al. J. Med. Chem. 1991, 34, 2871-76).

[0011] As mentioned above, the effect of deuteration on drug metabolism is not easy to predict. Obtunib and its metabolites in vivo have the defect of hepatotoxicity risk, which has caused great clinical concerns. Hepatotoxicity is not only related to the chemical structure, but also closely related to the clinical dosage.

[0012] The present invention determines chemical entities different from orelabrutinib by actually preparing and testing various deuterated orelabrutinibs, and obtains a class of deuterated drugs with good pharmacokinetic properties, reduced dosage and / or reduced toxic side effects by deuterating orelabrutinib. Summary of the invention

[0013] The purpose of the present invention is to provide a novel pyridine amide compound having BTK inhibitory activity and use thereof.

[0014] In the first aspect of the present invention, there is provided a pyridine amide compound of formula I, its tautomer, its crystal form, its salt, its hydrate or solvate

[0015]

[0016] in:

[0017] 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 are 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 them is deuterium.

[0018] In a preferred embodiment, the compound is the following compound, wherein: R 1 is hydrogen or deuterium; R 2 is hydrogen or deuterium; R 3 is hydrogen or deuterium; R 4 , R 5 , R 6 , R7 , R 8 , R 9 , R 10 , R 11 , R 12 , R 13 , R 14 , R 15 and R 16 At the same time, R 17 , R 18 and R 19 At the same time, the condition is that R 1 When it is hydrogen, R 2 and R 3 The compounds are the following compounds P001, P002, P003, P004 and P005.

[0019]

[0020] In another preferred embodiment, the compound is the compound described below, wherein: R 1 is hydrogen or deuterium; R 2 is hydrogen or deuterium; R 3 is hydrogen or deuterium; R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 11 , R 12 , R 13 , R 14 , R 15 and R 16 At the same time, R 17 , R 18 and R 19 is hydrogen or deuterium at the same time, provided that R 1 When it is deuterium, R 17 , R 18 and R 19 The compounds are the following compounds P006, P007, P008, P009, P010, P011, P012, P013, P014 and P015.

[0021]

[0022] In the second aspect of the present invention, a method for preparing a pharmaceutical composition is provided, comprising the steps of: mixing a pharmaceutically acceptable carrier with the compound described in the first aspect of the present invention, its tautomer, its crystalline form, its salt, its hydrate or solvate, to form a pharmaceutical composition.

[0023] In the third aspect of the present invention, a pharmaceutical composition is provided, which contains a pharmaceutically acceptable carrier and the compound described in the first aspect of the present invention, its tautomer, its crystal form, its salt, its hydrate or solvate.

[0024] In the fourth aspect of the present invention, provided is the use of the compound described in the first aspect of the present invention, its tautomer, its crystal form, its salt, its hydrate or solvate, which is used to prepare a pharmaceutical composition for inhibiting BTK.

[0025] In another preferred embodiment, the pharmaceutical composition is used to prevent and / or treat diseases related to BTK.

[0026] In another preferred embodiment, the pharmaceutical composition is used to prevent and / or treat a BTK-related disease, such as an allergic reaction, an autoimmune disease, an inflammatory disease, a thromboembolic disease or a cancer.

[0027] In another preferred embodiment, the pharmaceutical composition is used to treat rheumatoid arthritis, psoriatic arthritis, infectious arthritis, progressive chronic arthritis, teratogenic arthritis, osteoarthritis, traumatic arthritis, gouty arthritis, Reiter's syndrome, polychondritis, acute synovitis, spondylitis, glomerulonephritis with nephrotic syndrome, glomerulonephritis without nephrotic syndrome, autoimmune blood system disorders, hemolytic anemia, aplastic anemia, idiopathic thrombocytopenia, neutropenia, autoimmune gastritis, autoimmune inflammatory bowel disease, ulcerative colitis, Crohn's disease, host-versus-graft disease, allograft rejection, chronic thyroiditis, Graves' disease, scleroderma, type I diabetes, type II diabetes, acute active hepatitis, chronic active hepatitis, pancreatitis, primary biliary cirrhosis, myasthenia gravis, multiple sclerosis, systemic lupus erythematosus, psoriasis disease, atopic dermatitis, contact dermatitis, wet itching, sunburn, vasculitis, Behcet's disease, chronic renal insufficiency, Stevens-Johnson syndrome, inflammatory pain, idiopathic steatorrhea, cachexia, sarcoidosis, Guillain-Barré syndrome, uveitis, conjunctivitis, keratoconjunctivitis, otitis media, periodontal disease, pulmonary interstitial fibrosis, asthma, bronchitis, rhinitis, sinusitis, pneumoconiosis, pulmonary insufficiency syndrome, emphysema, pulmonary fibrosis, sand lung, chronic inflammatory lung disease, chronic obstructive pulmonary disease, proliferative diseases, non-Hodgkin's lymphoma, diffuse large B-cell lymphoma (DLBCL), mantle cell lymphoma (MCL), B-cell chronic lymphocytic leukemia, acute lymphoblastic leukemia, acute lymphoblastic leukemia with mature B cells, B-cell lymphoma caused by chronic active B-cell receptor signaling, and bone disease associated with multiple myeloma.

[0028] In another preferred embodiment, the pharmaceutical composition is used to treat chronic lymphocytic lymphoma, non-Hodgkin's lymphoma, diffuse large B-cell lymphoma, mantle cell lymphoma, follicular lymphoma and chronic lymphocytic leukemia.

[0029] As used herein, "deuterated" refers to a compound or group in which one or more hydrogen atoms are replaced by deuterium. Deuterated can be monosubstituted, disubstituted, polysubstituted or fully substituted.

[0030] 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%), preferably greater than 50%, more preferably greater than 85%, more preferably greater than 95%, more preferably greater than 99%, and more preferably greater than 99.5%.

[0031] In another preferred embodiment, the compound of formula I contains at least 1 or 3 deuterium atoms, more preferably 5 or 8 deuterium atoms.

[0032] As used herein, the term "compound of the present invention" refers to a compound represented by Formula I. The term also includes tautomers of the compound of Formula I, crystal forms thereof, salts thereof, hydrates thereof, or solvates thereof.

[0033] As used herein, the term "pharmaceutically acceptable salt" refers to a salt formed by a compound of the present invention and an acid or base that is suitable for use as a drug. Pharmaceutically acceptable salts include inorganic salts and organic salts. A preferred class of salts is a salt formed by a compound of the present invention and an acid. Suitable acids 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, 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, benzenesulfonic acid, and acidic amino acids such as aspartic acid and glutamic acid.

[0034] The compounds of the present invention or pharmaceutically acceptable derivatives thereof may also be administered at the same time, before or after administering one or more other therapeutic agents. This combination therapy includes administering a single pharmaceutical administration preparation containing the compounds of the present invention and one or more other active agents, as well as administering a separate pharmaceutical administration preparation of the compounds of the present invention and each active agent. For example, the compounds of the present invention and another active agent can be administered to the patient together with a single oral administration composition (e.g., tablets or capsules), or each agent is administered with a separate oral administration preparation. In the case of using a separate administration preparation, the compounds of the present invention and one or more additional active agents can be administered substantially at the same time (i.e., simultaneously) or at a separate staggered time (i.e., successively); combination therapy should be understood to include all of these schemes.

[0035] The dosage of the pharmaceutical combination of the present invention will be adjusted according to the disease state, route of administration, age or weight of the patient. For oral administration in adults, it is usually 0.2-35 mg / kg / day, preferably 0.8-20 mg / kg / day. The suitable dosage of the present invention needs to be set considering the patient's age, weight, condition, route of administration, etc., and oral administration is usually 0.1-35 mg / kg / day, preferably in the range of 0.5-15 mg / kg / day.

[0036] The positive and progressive effects of the present invention are:

[0037] (1) The compounds of the present invention have good selective BTK inhibitory effects and can be effectively used for treating diseases associated with BTK.

[0038] (2) The compound of the present invention has good selectivity in inhibiting B cell activation and is effectively used as a B cell activation inhibitor.

[0039] (3) The deuterated pyridine amide compounds and pharmaceutically acceptable salts thereof of the present invention have low hepatotoxicity, good pharmacokinetic properties, reduced dosage and / or reduced toxic side effects, and better drugability. Compared with orelabrutinib, they have significantly better pharmacokinetic and / or pharmacodynamic properties and / or safety properties, and are more suitable for the preparation and treatment of diseases related to BTK.

[0040] The preparation method of the compound of formula I of the present invention is described in more detail below, but these specific methods do not constitute any limitation to the present invention. The compounds of the present invention can also be conveniently prepared by optionally combining various synthetic methods described in this specification or known in the art, and such a combination can be easily carried out by a technician in the field to which the present invention belongs.

[0041] The preparation methods of the non-deuterated pyridine amide compounds and their physiologically compatible salts used in the present invention are known. The corresponding deuterated pyridine amide compounds can be synthesized using the corresponding deuterated starting compounds as raw materials and in the same way.

[0042] Taking compound P001 as an example, a preferred preparation process is as follows:

[0043]

[0044] The specific synthesis method is described in Example 1.

[0045] The present invention is further described below in conjunction with specific examples. These examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. The experimental methods in the following examples without specifying specific conditions are usually carried out under conventional conditions or under conditions recommended by the manufacturer.

[0046] Example 1: Synthesis of Compound P001

[0047] Synthetic route

[0048]

[0049] Step 1: Synthesis of compound P102

[0050]

[0051] 2,6-Dichloronicotinonitrile (Compound P101) (10 mmol) was added to a mixed solution of concentrated sulfuric acid (10 ml) and water (2 ml), heated to 90°C, and stirred for 1 hour. After cooling to room temperature, the reaction mixture was placed in an ice-water bath and the pH value was adjusted to 8 with aqueous ammonia. The filter cake was filtered, washed with water, and dried to obtain Compound P102.

[0052] Step 2: Synthesis of compound P104

[0053]

[0054] Compound P102 (3.0 mmol), 4-phenoxyphenylboronic acid-d5 (compound P103) (3.0 mmol), tris(dibenzylideneacetone)dipalladium (0.55 mmol), and cesium carbonate (6.0 mmol) were added to a mixed solution of 1,4-dioxane (60 ml) and water (12 ml), and then heated to 120°C and refluxed with stirring for 16 hours. The mixture was concentrated under reduced pressure, and the residue was purified by a silica gel column with dichloromethane / methanol (150 / 1) as the eluent, and the solvent was removed to obtain compound P104.

[0055] Step 3: Synthesis of compound P106

[0056]

[0057] Compound P104 (0.50 mmol), (1-(tert-butoxycarbonyl)-1,2,3,6-tetrahydropyridin-4-yl)boric acid (compound P105) (0.75 mmol), tetrakis(triphenylphosphine)palladium (0.1 mmol) and potassium carbonate (1.5 mmol) were added to a mixed solution of ethylene glycol dimethyl ether (10 ml) and water (2 ml), heated to 90°C, and stirred for 5 hours. The mixture was cooled to room temperature and concentrated under reduced pressure. The residue was purified by silica gel column with dichloromethane / methanol (700 / 1) as the eluent, and the solvent was removed to obtain compound P106.

[0058] Step 4: Synthesis of compound P107

[0059]

[0060] Compound P106 (0.45 mmol) and palladium carbon (10 mg) were added to ethyl acetate (10 ml), and the mixture was stirred at room temperature for 16 hours under a hydrogen atmosphere with no oxygen. The mixture was concentrated under reduced pressure, and the residue was purified by silica gel column with dichloromethane / methanol (70 / 1) as the eluent, and the solvent was removed to obtain compound P107.

[0061] Step 5: Synthesis of compound P108

[0062]

[0063] Compound P107 and trifluoroacetic acid (2 ml) were added to dichloromethane (5 ml) and stirred at room temperature for 1 hour. The mixture was concentrated under reduced pressure, and the residue was dissolved in dichloromethane and washed with saturated sodium bicarbonate solution. The organic phase was dried over anhydrous sodium sulfate, filtered to remove the desiccant, and concentrated under reduced pressure. The residue was purified by silica gel column with dichloromethane / methanol (70 / 1 to 5 / 1) as eluent. The target product fractions were combined and the solvent was removed to obtain compound P108.

[0064] Step 6: Synthesis of compound P001

[0065]

[0066] Compound P108 (0.40 mmol), acryloyl chloride-d3 (compound P109) (0.56 mmol) and triethylamine (0.85 mmol) were added to dichloromethane (6 ml) and stirred at 0°C for 1 hour. The reaction was quenched with water (10 ml), diluted with ethyl acetate (30 ml), and washed with water (20 ml × 2) and saturated brine (20 ml × 2) in sequence. The organic phase was dried over anhydrous sodium sulfate, filtered to remove the desiccant, concentrated under reduced pressure, and the residue was purified by silica gel column. The eluent was dichloromethane / methanol (20 / 1), and the solvent was removed to obtain compound P001. The H NMR spectrum of compound P001 is: 1 H-NMR(CDCl3)δ8.0(1H),7.8-7.6(2H),7.2-6.9(3H),5.8(1H),5.5(1H),4 .8(1H),4.1(1H),3.2(1H),3.1(1H),2.8(1H),2.0-1.9(2H),1.8-1.6(2H).

[0067] Example 2: Synthesis of Compound P002

[0068] For the synthesis of compound P002, the compound P103 in step 2 of "Example 1: Synthesis of compound P001" was replaced with compound P202. The remaining steps were performed in the same manner as in "Example 1: Synthesis of compound P001" to obtain compound P002. The H NMR spectrum of compound P002 was: 1 H-NMR(CDCl3)δ8.0(1H),7.8-7.6(2H),7.5-7.3(2H),7.2-6.9(5H),5.8(1H),5.5 (1H),4.8(1H),4.1(1H),3.2(1H),3.1(1H),2.8(1H),2.0-1.9(2H),1.8-1.6(2H).

[0069]

[0070] Example 3: Synthesis of Compound P003

[0071] For the synthesis of compound P003, the compound P103 in step 2 of "Example 1: Synthesis of compound P001" was replaced with compound P203. The remaining steps were the same as those in "Example 1: Synthesis of compound P001" to obtain compound P003. The H NMR spectrum of compound P003 was: 1H-NMR(CDCl3)δ8.0(1H),7.8-7.6(2H),7.5-7.3(2H),7.2-6.9(3H),5.8(1H),5.5 (1H),4.8(1H),4.1(1H),3.2(1H),3.1(1H),2.8(1H),2.0-1.9(2H),1.8-1.6(2H).

[0072]

[0073] Example 4: Synthesis of Compound P004

[0074] For the synthesis of compound P004, the compound P103 in step 2 of "Example 1: Synthesis of compound P001" was replaced with compound P204. The remaining steps were the same as those in "Example 1: Synthesis of compound P001" to obtain compound P004. The H NMR spectrum of compound P004 was: 1 H-NMR(CDCl3)δ8.0(1H),7.8-7.6(2H),7.2-6.9(5H),5.8(1H),5.5(1H),4 .8(1H),4.1(1H),3.2(1H),3.1(1H),2.8(1H),2.0-1.9(2H),1.8-1.6(2H).

[0075]

[0076] Example 5: Synthesis of Compound P005

[0077] For the synthesis of compound P005, the compound P103 in step 2 of "Example 1: Synthesis of compound P001" was replaced with compound P205. The remaining steps were the same as those in "Example 1: Synthesis of compound P001" to obtain compound P005. The H NMR spectrum of compound P005 was: 1 H-NMR(CDCl3)δ8.0(1H),7.8-7.6(2H),7.5-7.3(2H),7.2-6.9(6H),5.8(1H),5.5 (1H),4.8(1H),4.1(1H),3.2(1H),3.1(1H),2.8(1H),2.0-1.9(2H),1.8-1.6(2H).

[0078]

[0079] Example 6: Synthesis of Compound P006

[0080] For the synthesis of compound P006, the compound P109 in step 6 of "Example 1: Synthesis of compound P001" was replaced with compound P206. The remaining steps were performed in the same manner as in "Example 1: Synthesis of compound P001" to obtain compound P006. The H NMR spectrum of compound P006 was: 1 H-NMR(CDCl3)δ8.0(1H),7.8-7.6(2H),7.2-6.9(3H),6.6(1H),6.3(1H),5.8(1H),5.7(1 H),5.5(1H),4.8(1H),4.1(1H),3.2(1H),3.1(1H),2.8(1H),2.0-1.9(2H),1.8-1.6(2H).

[0081]

[0082] Example 7: Synthesis of Compound P007

[0083] For the synthesis of compound P007, the compound P103 in step 2 of "Example 6: Synthesis of compound P006" was replaced with compound P202. The remaining steps were the same as those in "Example 6: Synthesis of compound P006" to obtain compound P007. The H NMR spectrum of compound P007 is: 1 H-NMR(CDCl3)δ8.0(1H),7.8-7.6(2H),7.5-7.3(2H),7.2-6.9(5H),6.6(1H),6.3(1H),5.8(1H) ,5.7(1H),5.5(1H),4.8(1H),4.1(1H),3.2(1H),3.1(1H),2.8(1H),2.0-1.9(2H),1.8-1.6(2H).

[0084]

[0085] Example 8: Synthesis of Compound P008

[0086] For the synthesis of compound P008, the compound P103 in step 2 of "Example 6: Synthesis of compound P006" was replaced with compound P203. The remaining steps were the same as those in "Example 6: Synthesis of compound P006" to obtain compound P008. The H NMR spectrum of compound P008 was: 1H-NMR(CDCl3)δ8.0(1H),7.8-7.6(2H),7.5-7.3(2H),7.2-6.9(3H),6.6(1H),6.3(1H),5.8(1H) ,5.7(1H),5.5(1H),4.8(1H),4.1(1H),3.2(1H),3.1(1H),2.8(1H),2.0-1.9(2H),1.8-1.6(2H).

[0087]

[0088] Example 9: Synthesis of Compound P009

[0089] For the synthesis of compound P009, the compound P103 in step 2 of "Example 6: Synthesis of compound P006" was replaced with compound P204. The remaining steps were the same as those in "Example 6: Synthesis of compound P006" to obtain compound P009. The H NMR spectrum of compound P009 is: 1 H-NMR(CDCl3)δ8.0(1H),7.8-7.6(2H),7.2-6.9(5H),6.6(1H),6.3(1H),5.8(1H),5.7(1 H),5.5(1H),4.8(1H),4.1(1H),3.2(1H),3.1(1H),2.8(1H),2.0-1.9(2H),1.8-1.6(2H).

[0090]

[0091] Example 10: Synthesis of Compound P010

[0092] For the synthesis of compound P010, the compound P103 in step 2 of "Example 1: Synthesis of compound P001" was replaced with compound P207. The remaining steps were the same as those in "Example 1: Synthesis of compound P001" to obtain compound P010. The H NMR spectrum of compound P010 was: 1 H-NMR(CDCl3)δ8.0(1H),7.8-7.6(2H),7.2-6.9(4H),5.8(1H),5.5(1H),4 .8(1H),4.1(1H),3.2(1H),3.1(1H),2.8(1H),2.0-1.9(2H),1.8-1.6(2H).

[0093]

[0094] Example 11: Synthesis of Compound P011

[0095] For the synthesis of compound P011, the compound P103 in step 2 of "Example 1: Synthesis of compound P001" was replaced with compound P208. The remaining steps were the same as those in "Example 1: Synthesis of compound P001" to obtain compound P011. The H NMR spectrum of compound P011 is: 1 H-NMR(CDCl3)δ8.0(1H),7.8-7.6(2H),7.5-7.3(2H),7.2-6.9(4H),5.8(1H),5.5 (1H),4.8(1H),4.1(1H),3.2(1H),3.1(1H),2.8(1H),2.0-1.9(2H),1.8-1.6(2H).

[0096]

[0097] Example 12: Synthesis of Compound P012

[0098] For the synthesis of compound P012, the compound P103 in step 2 of "Example 1: Synthesis of compound P001" was replaced with compound P209. The remaining steps were the same as those in "Example 1: Synthesis of compound P001" to obtain compound P012. The H NMR spectrum of compound P012 is: 1 H-NMR(CDCl3)δ8.0(1H),7.8-7.6(2H),7.2-6.9(6H),5.8(1H),5.5(1H),4 .8(1H),4.1(1H),3.2(1H),3.1(1H),2.8(1H),2.0-1.9(2H),1.8-1.6(2H).

[0099]

[0100] Example 13: Synthesis of Compound P013

[0101] For the synthesis of compound P013, the compound P103 in step 2 of "Example 6: Synthesis of compound P006" was replaced with compound P207. The remaining steps were the same as those in "Example 6: Synthesis of compound P006" to obtain compound P013. The H NMR spectrum of compound P013 is: 1 H-NMR(CDCl3)δ8.0(1H),7.8-7.6(2H),7.2-6.9(4H),6.6(1H),6.3(1H),5.8(1H),5.7(1 H),5.5(1H),4.8(1H),4.1(1H),3.2(1H),3.1(1H),2.8(1H),2.0-1.9(2H),1.8-1.6(2H).

[0102]

[0103] Example 14: Synthesis of Compound P014

[0104] For the synthesis of compound P014, the compound P103 in step 2 of "Example 6: Synthesis of compound P006" was replaced with compound P208. The remaining steps were the same as those in "Example 6: Synthesis of compound P006" to obtain compound P014. The H NMR spectrum of compound P014 is: 1 H-NMR(CDCl3)δ8.0(1H),7.8-7.6(2H),7.5-7.3(2H),7.2-6.9(4H),6.6(1H),6.3(1H),5.8(1H) ,5.7(1H),5.5(1H),4.8(1H),4.1(1H),3.2(1H),3.1(1H),2.8(1H),2.0-1.9(2H),1.8-1.6(2H).

[0105]

[0106] Example 15: Synthesis of Compound P015

[0107] For the synthesis of compound P015, the compound P103 in step 2 of "Example 6: Synthesis of compound P006" was replaced with compound P209. The remaining steps were the same as those in "Example 6: Synthesis of compound P006" to obtain compound P015. The H NMR spectrum of compound P015 is: 1 H-NMR(CDCl3)δ8.0(1H),7.8-7.6(2H),7.2-6.9(6H),6.6(1H),6.3(1H),5.8(1H),5.7(1 H),5.5(1H),4.8(1H),4.1(1H),3.2(1H),3.1(1H),2.8(1H),2.0-1.9(2H),1.8-1.6(2H).

[0108]

[0109] Example 16: Synthesis of Compound P103

[0110]

[0111] Step 7: Synthesis of compound P403

[0112] Compound P401 (11.8 g) was dissolved in anhydrous tetrahydrofuran (120 ml), stirred, and sodium hydride (12.1 g) was slowly added in batches, and then 1-bromo-4-iodobenzene (compound P402) (39.1 g) was added in batches. The reaction was allowed to react at room temperature for 15 hours, and the reactant was filtered, the filtrate was dried by rotary evaporation, and dichloromethane was added to dissolve. The mixture was passed through a silica gel column and eluted with petroleum ether: ethyl acetate (1:5) to obtain compound P403 (12.7 g).

[0113] Step 8: Synthesis of compound P103

[0114] Compound P403 (5.2 g) was dissolved in dry THF (100 ml), and reacted at -78 °C for 30 min under N2 protection. n-Butyl lithium (1.9 g) was slowly added dropwise, and the temperature was kept at -78 °C for 3 h after the addition was completed. Triisopropyl borate (4.3 g) was then slowly added dropwise, and the temperature was kept at -78 °C for 2 h after the addition was completed. The temperature was slowly raised to room temperature and the reaction was continued for about 15 hr. After the reaction was completed as monitored by TLC, the reaction solution was slowly quenched with water, extracted and concentrated to obtain compound P103 (3.0 g).

[0115] Example 17: Synthesis of Compound P203

[0116] The synthesis of compound P203 was carried out according to "Example 16: Synthesis of compound P103", except that in step 7, compound P401 was replaced by compound P402, and the remaining steps were carried out in the same manner as "Example 16: Synthesis of compound P103" to obtain compound P203.

[0117]

[0118] Example 18: Synthesis of Compound P204

[0119] The synthesis of compound P204 was carried out according to "Example 16: Synthesis of compound P103", except that in step 7, compound P401 was replaced by compound P403, and the remaining steps were carried out in the same manner as "Example 16: Synthesis of compound P103" to obtain compound P204.

[0120]

[0121] Example 19: Synthesis of Compound P202

[0122] The synthesis of compound P202 was carried out according to "Example 16: Synthesis of compound P103", except that in step 7, compound P401 was replaced by compound P404, and the remaining steps were carried out in the same manner as "Example 16: Synthesis of compound P103" to obtain compound P202.

[0123]

[0124] Example 20: Synthesis of Compound P207

[0125] The synthesis of compound P207 was carried out according to "Example 16: Synthesis of compound P103", except that in step 7, compound P401 was replaced by compound P405, and the remaining steps were carried out in the same manner as "Example 16: Synthesis of compound P103" to obtain compound P207.

[0126]

[0127] Example 21: Synthesis of Compound P208

[0128] The synthesis of compound P208 was carried out according to "Example 16: Synthesis of compound P103", except that in step 7, compound P401 was replaced by compound P406, and the remaining steps were carried out in the same manner as "Example 16: Synthesis of compound P103" to obtain compound P208.

[0129]

[0130] Example 22: Synthesis of Compound P209

[0131] The synthesis of compound P209 was carried out according to "Example 16: Synthesis of compound P103", except that in step 7, compound P401 was replaced by compound P407. The remaining steps were carried out in the same manner as "Example 16: Synthesis of compound P103" to obtain compound P209.

[0132]

[0133] Example 23: Pharmacokinetics in rats

[0134] Thirty male Sprague-Dawley rats, aged 6-9 weeks and weighing about 220 g, were divided into 5 groups (orelabrutinib group, compound P001 group, compound P002 group, compound P005 group and compound P009 group), with 6 rats in each group. Orelabrutinib, compound P001, compound P002, compound P005 and compound P009 were given a single oral gavage at a dose of 10 mg / kg according to the grouping, and their pharmacokinetic differences were compared.

[0135] Rats were fasted 12 hours before administration. The administration solution was prepared with 0.5% sodium carboxymethylcellulose (CMC-Na). Blood was collected from the orbital venous plexus at 0.25 hours, 0.5 hours, 1 hour, 2 hours, 4 hours, 6 hours, 8 hours, 12 hours, 16 hours and 24 hours after administration. After plasma was separated from the blood sample, the plasma was stored in a -80°C refrigerator for future use. An LC-MS / MS analysis method was established to measure the plasma samples.

[0136] The test results show that compared with orbutinib, the elimination half-life T 1 / 2 and / or area under the curve AUC and / or maximum plasma concentration C max An increase of more than 50%.

[0137] From the present results, it can be seen that compared with obeticholic acid, compound P001, compound P002, compound P005 and / or compound P009 of the present invention have better pharmacokinetic properties in rats, indicating better pharmacodynamics and therapeutic effects.

[0138] Example 24: Pharmacokinetics in rats

[0139] 24 male Sprague-Dawley rats, 6-9 weeks old, weighing about 220g, were divided into 4 groups (obbutinib group, compound P006 group, compound P007 group, compound P012 group), 6 rats in each group. Obbutinib, compound P006, compound P007 and compound P012 were given a single oral gavage at a dose of 10 mg / kg according to the grouping, and their pharmacokinetic differences were compared. Rats were fasted 12 hours before administration. The dosing solution was prepared with 0.5% sodium carboxymethyl cellulose (CMC-Na). Blood was collected from the orbital venous plexus at 0.25 hours, 0.5 hours, 1 hour, 2 hours, 4 hours, 6 hours, 8 hours, 12 hours, 16 hours and 24 hours after administration. After plasma was separated from the blood sample, the plasma was stored in a -80°C refrigerator for use. LC-MS / MS analysis method was established to measure plasma samples.

[0140] The experimental results show that compared with orbutinib, the elimination half-life of compound P006, compound P007 and compound P012 is 1 / 2 and / or area under the curve AUC and / or maximum plasma concentration C max An increase of more than 40%.

[0141] From the present results, it can be seen that compared with orelabrutinib, compound P006, compound P007 and / or compound P012 of the present invention have better pharmacokinetic properties in rats, indicating better pharmacodynamics and therapeutic effects.

[0142] Example 25: Determination of BTK inhibitory activity

[0143] Using ADP-Glo TM The kit is used to measure the effect of the compound of the present invention on the activity of BTK. The experimental method is as follows:

[0144] ADP is the product of kinase reaction, and kinase activity can usually be detected by detecting the amount of ADP generated. ADP-Glo ​​developed by Promega TM The kit measures the in vitro activity of BTK by detecting the level of ADP produced in the kinase reaction. In the kinase detection experiment, the kinase consumes ATP to phosphorylate the substrate and produces ADP. Then the ADP-Glo ​​reagent is added to terminate the kinase reaction and completely consume the remaining ATP. The kinase detection reagent is then added to convert the generated ADP into new ATP. The luciferase in the detection reagent can catalyze luciferin with the participation of ATP and O2 to produce a light signal, thereby converting the chemical signal into a light signal, and the intensity of the light signal is positively correlated with the amount of ADP produced in the kinase reaction, so that the activity of the kinase BTK can be quantitatively detected.

[0145] All detection experiments were carried out at a constant temperature of 23°C, using Corning 3674 white 384-well detection plates, kinase BTK (Invitrogen), kinase substrates peptide (4:l Glu, Tyr) (Signal Chem) and ATP (Sigma), and the light signal was read using the microplate reader EnVision (Perkin Elmer). The detection buffer included 40mM Tris-HCl (pH 7.5), 10mM MgCl2 (Sigma), 2mM MnCl2 (Sigma), 0.05mM DTT (Sigma) and 0.01% BSA (Sigma); the kinase BTK was prepared into a kinase reaction solution with a concentration of 1.3ng / μL using the detection buffer; the substrate reaction solution included 0.25mg / mL peptide substrate and 60μM ATP.

[0146] The compound of the present invention was diluted with DMSO to a 0.5 mM solution, and then three-fold gradient dilution was performed with DMSO to a minimum concentration of 0.025 μM. 50 nL of compound solution of a series of concentrations and 2.5 μL of kinase reaction solution were first added to a 384-well plate using Echo555, mixed evenly, and incubated at room temperature in the dark for 30 minutes; then 2.5 μL of 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 ADP-Glo ​​was added TM The reagent terminated the reaction, mixed evenly and placed at room temperature for 40 minutes; finally, 10 μL of kinase detection reagent was added, placed at room temperature away from light for 30 minutes, and then the value was read on Envision.

[0147] The inhibition percentage was calculated according to the following formula:

[0148] Inhibition % = [1-(RLU 化合物 -RLU min ) / (RLUmax -RLU min )]×100

[0149] 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 kinase BTK, RLU max The IC values ​​of the compounds were calculated by using the XLfit program in Excel. 50 value.

[0150] Table 1: IC values ​​of compounds of the present invention 50 value

[0151] Compound <![CDATA[IC 50 (nM)]]> Compound <![CDATA[IC 50 (nM)]]> P001 1.4 P002 2.7 P006 2.4 P007 1.9 P008 2.6 P012 3.1 P015 3.4 Orelabrutinib 3.9

[0152] It can be seen from the present results that the compounds of the present invention have a significant inhibitory effect on BTK.

[0153] Example 26: Comparative study of liver toxicity in mice

[0154] (1) Experimental animals

[0155] Thirty-two adult male ICR mice with a body weight of (25±2 g) were selected, and all mice were allowed to freely access water and feed, and maintained under a day-night cycle at a temperature of 25±2° C. and a relative humidity of 50±10%.

[0156] (2) Animal grouping and drug administration

[0157] 32 male ICR mice were divided into four groups, 8 mice in each group, namely normal control group, model group, model + example compound group and model + Orelabrutinib group. The model + example compound group was intragastrically administered with the example compound once a day at a dose (50 mg / kg); the model + Orelabrutinib group was intragastrically administered with Orelabrutinib once a day at a dose (50 mg / kg), respectively, for 8-16 weeks, and the normal control group and the model group were intragastrically administered with an equal volume of purified water. Food was cut off after the last administration, and 1 hour later, the model group, the model + example compound group and the model + Orelabrutinib group mice were intraperitoneally injected with 250 mg / kg of acetaminophen (APAP) saline solution, respectively. After 24 hours of modeling, blood was collected from the eyeballs of mice in each group in turn, and the serum was separated by centrifugation at 3000r / min for 10 minutes, and stored at 4°C for later use; the liver and spleen were quickly dissected. After washing with 4°C saline, the liver was dried with filter paper and weighed. Part of the liver was fixed in 10% formaldehyde solution for slicing, and the remaining liver was stored in a -80°C low-temperature refrigerator.

[0158] (3) Determination of biochemical indicators in the liver:

[0159] Weigh part of the liver, add 9 times the volume of ice saline, use a tissue homogenizer to make 10% liver tissue homogenate, centrifuge and take the supernatant. Spot the plate according to the kit method, measure the OD value at 450nm, and calculate the MDA content and GSH activity in the liver according to the formula.

[0160] (4) Data processing

[0161] The experimental data were expressed as mean ± standard deviation (±s) and analyzed using SPSS22.0 statistical software. One-way analysis of variance was used to compare the differences between the groups. P < 0.05 was considered a significant difference.

[0162] (5) Effects of the compounds in the present examples on lipid peroxidation in liver tissue of mice with APAP liver injury

[0163] Compared with the normal control group, the MDA content in the liver tissue homogenate of the mice in the model group increased significantly, and the GSH level decreased significantly (P<0.05), which caused the accumulation of lipid peroxidation products in the mice and reduced the antioxidant metabolism level; compared with the model group, the MDA content and GSH level of the model + example compound group did not change significantly (P>0.05); compared with the model group, the MDA content of the model + Orelabrutinib 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 significant effect on lipid peroxidation caused by APAP, while Orelabrutinib (50 mg / kg) had an effect on lipid peroxidation caused by APAP, suggesting that the liver toxicity of the example compound of the present application was less than that of Orelabrutinib in mice. The results are shown in Table 2.

[0164] Table 2. Effects on lipid peroxidation in liver tissue of mice with APAP liver injury

[0165] Grouping MDA (nmol / mg) GSH (μmol / L) Normal control group A- B- Model Group A+ B+ Model + Example Compound Group A+ B+ Model+Orelabrutinib group A++ B++

[0166] Marking (grade): A+ is 2.4-3.5; A- is 1.5-2.2; A++ is 3.8-5.2; B+ is 28-39; B- is 42-52; B++ is 16-27.

[0167] Conclusion: The compound of the present invention (50 mg / kg) has no significant effect on lipid peroxidation induced by APAP, while Orelabrutinib (50 mg / kg) has an effect on lipid peroxidation induced by APAP, indicating that the compound of the present invention is less toxic to the mouse liver than Orelabrutinib.

[0168] Finally, it is necessary to explain that the specific embodiments of the present invention are described in detail above, but they are only examples, 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, the equalization changes and modifications made without departing from the spirit and scope of the present invention should be included in the scope of the present invention.

Claims

1. A pyridine amide compound, a tautomer or a salt thereof as represented by formula I, in: 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 are each independently selected from hydrogen or deuterium, 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 them is deuterium; The compound is selected from the following structures:

2. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and the pyridinamide compound, tautomer or salt thereof according to claim 1.

3. Use of the pyridine amide compound, tautomer or salt thereof according to claim 1 in the preparation of a BTK inhibitor.

4. Use of the pharmaceutical composition according to claim 2 in the preparation of drugs for treating and / or preventing diseases associated with BTK.

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

6. Use of the pyridinamide compound, tautomer or salt thereof according to claim 1 in the preparation of a medicament for treating a BTK-mediated disease, wherein the BTK-mediated disease is selected from the group consisting of rheumatoid arthritis, psoriatic arthritis, infectious arthritis, progressive chronic arthritis, teratogenic arthritis, osteoarthritis, traumatic arthritis, gouty arthritis, Reiter's syndrome, polychondritis, acute synovitis, spondylitis, glomerulonephritis with nephrotic syndrome, glomerulonephritis without nephrotic syndrome, autoimmune blood system disease, hemolytic anemia, aplastic anemia, idiopathic thrombocytopenia, neutropenia, autoimmune gastritis, autoimmune inflammatory bowel disease, ulcerative colitis, Crohn's disease, host versus graft disease, allograft rejection, chronic thyroiditis, Graves' disease, scleroderma, type I diabetes, type II diabetes, acute active hepatitis, chronic active hepatitis, pancreatitis, primary biliary tract infection, Juicy cirrhosis, myasthenia gravis, multiple sclerosis, systemic lupus erythematosus, psoriasis, atopic dermatitis, contact dermatitis, wet itching, sunburn, vasculitis, Behcet's disease, chronic renal insufficiency, Stevens-Johnson syndrome, inflammatory pain, idiopathic steatorrhea, cachexia, sarcoidosis, Guillain-Barré syndrome, uveitis, conjunctivitis, keratoconjunctivitis, otitis media, periodontal disease, pulmonary fibrosis, asthma, bronchitis Pneumoconiosis, rhinitis, sinusitis, pneumoconiosis, pulmonary insufficiency syndrome, emphysema, pulmonary fibrosis, silicosis, chronic inflammatory lung disease, chronic obstructive pulmonary disease, proliferative diseases, non-Hodgkin's lymphoma, diffuse large B-cell lymphoma, mantle cell lymphoma, B-cell chronic lymphocytic leukemia, acute lymphoblastic leukemia, acute lymphoblastic leukemia with mature B cells, B-cell lymphoma caused by chronic active B-cell receptor signaling, and bone disease associated with multiple myeloma.

7. The use according to claim 6, wherein the BTK-mediated disorder is a B-cell proliferative disorder.

8. The use according to claim 7, wherein the B-cell proliferative disorder is selected from the group consisting of chronic lymphocytic lymphoma, non-Hodgkin's lymphoma, diffuse large B-cell lymphoma, mantle cell lymphoma, follicular lymphoma and chronic lymphocytic leukemia.

Images