Deuterated aminopyridine derivative and pharmaceutical composition containing same

Abstract

The invention relates to a deuterated aminopyridine derivative shown as a formula I, an optical isomer thereof or a mixture thereof, a crystal form thereof, a salt thereof, a hydrate thereof or a solvate thereof. The invention also relates to a pharmaceutical composition containing a pharmaceutically acceptable carrier and the deuterated aminopyridine derivative, the optical isomer thereof or the mixture thereof, the crystal form thereof, the salt thereof, the hydrate thereof or the solvate thereof. The compound represented by the general formula I is used as a BTK inhibitor, is a prevention and / or treatment agent for BTK-related diseases, has low hepatotoxicity, and has a good treatment effect on allergic diseases, autoimmune diseases, inflammatory diseases, thromboembolic diseases or cancers.

Description

Technical Field

[0001] The invention belongs to the field of biomedicine, and specifically relates to deuterated aminopyridine derivatives and a pharmaceutical composition containing the compound. 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 that are independent of T cells, and significant reductions in sustained calcium signals during BCR stimulation. Evidence that BTK plays a role in allergic diseases and / or autoimmune diseases and / or inflammatory diseases has been determined 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 the production of autoantibodies). 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 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).

[0005] Furthermore, aberrant activation of BTK plays an important role in the pathogenesis of B-cell lymphomas, which means that inhibition of 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 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).

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

[0007] Tolebrutinib (SAR442168, PRN2246) is a potent, selective, orally active and blood-brain barrier-permeable Bruton's tyrosine kinase (BTK) inhibitor. It is an investigational brain-penetrating Bruton's tyrosine kinase (BTK) inhibitor and the first drug to complete a proof-of-concept study of BTK inhibitors for the treatment of multiple sclerosis (MS). It can produce the cerebrospinal fluid (CSF) concentrations required to target microglia and B lymphocytes. It is undergoing Phase III clinical trials for multiple sclerosis (MS) and myasthenia gravis (MG). However, cases of drug-induced liver injury 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 elimination half-life of Tolebrutinib in humans is1 / 2 Between 1.39-2.18 hours, it is eliminated very quickly (Owens TD et al. Clin Transl Sci. 2022; 15: 442–450). This suggests that a larger dose is needed to maintain a better therapeutic effect, but this also inevitably brings a greater risk of toxicity. In addition, Tolebrutinib has a short half-life and is eliminated very quickly, which also indicates to a certain extent that Tolebrutinib may be metabolized more quickly, and the metabolites produced may also increase the risk of toxicity. The poor absorption, distribution, metabolism and / or excretion (ADME) performance of some current drugs has hindered their wider use or limited their use in specific indications. For example, due to the short elimination half-life and rapid clearance of drugs in the body, a common solution is to administer 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, which will cause more severe side effects and increase treatment costs. Rapidly metabolized drugs may also expose patients to undesirable toxic or reactive metabolites.

[0009] Another ADME limitation affecting drugs is the formation of toxic or biologically reactive metabolites. Therefore, some patients receiving the drug may experience toxicity, or the safe dose of such a drug may be limited so that the patient receives a suboptimal amount of treatment. In some cases, changing the dosing interval or formulation method can help reduce clinical adverse reactions, but the frequent formation of such undesirable metabolites is inherent to compound metabolism.

[0010] A potential attractive strategy for improving drug metabolism performance is deuterium modification (modification). In this method, people attempt to slow down the metabolism of the drug, or by replacing one or more hydrogen atoms with deuterium atoms to reduce the formation of undesirable metabolites. 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 bond strength of the increase given by deuterium can positively affect the ADME performance of the drug, with the potential for improving drug effect, safety, and / or tolerability. Meanwhile, because the size and shape of deuterium are substantially equivalent to hydrogen, compared with the original chemical entity that only comprises hydrogen, it is expected that replacing hydrogen with deuterium will not affect the biochemical efficacy and selectivity of the drug.

[0011] 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")).

[0012] 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), and deteriorate its pharmacokinetic properties; on the other hand, hydrogen at certain positions on the drug molecule is not easily substituted with deuterium due to steric hindrance and other reasons.

[0013] 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).

[0014] In addition, deuteration can lead to metabolic switching. The concept of metabolic switching indicates that when a drug is encapsulated by a phase I metabolizing enzyme, it can briefly bind and rebind with the phase I metabolizing enzyme in various conformations before a chemical reaction (such as an oxidation reaction). Therefore, metabolic switching can potentially lead to different proportions of known metabolites and new metabolites. This new metabolic property can cause more or less toxicity. And lead to faster or slower drug clearance, thereby reducing or increasing the drug's in vivo exposure. Such changes caused by metabolic switching are unpredictable, and so far no sufficient a priori prediction has been made for any drug.

[0015] As mentioned above, the effect of deuterium modification on drug metabolism is unpredictable. Tolebrutinib and its metabolites in vivo have the disadvantage of hepatotoxicity risk, and clinical trials of Tolebrutinib have also shown hepatotoxicity, causing great clinical concerns. Hepatotoxicity is not only related to the chemical structure, but also closely related to the clinical dosage.

[0016] Therefore, in view of the shortcomings of existing technologies, we design new compounds to increase the exposure of new compounds in the body, reduce the dosage and / or frequency of administration, and reduce liver toxicity; and / or reduce the liver toxicity of its prototype or metabolites through structural modification, or reduce the generation of toxic metabolites, so as to achieve the purpose of reducing toxicity and increasing efficacy. Summary of the invention

[0017] The purpose of the present invention is to provide a novel class of compounds having BTK inhibitory activity and better pharmacodynamic properties and uses thereof.

[0018] 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.

[0019]

[0020] Where: 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 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.

[0021] In a preferred embodiment, the compound is a preferred compound selected from the following group:

[0022]

[0023] In another preferred embodiment, the compound is a preferred compound selected from the following group:

[0024]

[0025] 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 optical isomer or a mixture thereof, its crystal form, its salt, its hydrate or solvate, thereby forming a pharmaceutical composition.

[0026] 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 optical isomer or mixture thereof, its crystal form, its salt, its hydrate or solvate. In another preferred embodiment, the pharmaceutical composition is a capsule, tablet, injection, pill, powder or granule.

[0027] In the fourth aspect of the present invention, there is provided the use of the compound described in the first aspect of the present invention, its optical isomer or mixture thereof, its crystal form, its salt, its hydrate or solvate, which is used to prepare a pharmaceutical composition for inhibiting BTK. In another preferred embodiment, the pharmaceutical composition is used to prevent and / or treat diseases related to BTK.

[0028] In another preferred embodiment, the pharmaceutical composition is used to prevent and / or treat allergic diseases, autoimmune diseases, inflammatory diseases, thromboembolic diseases or cancer.

[0029] In another preferred embodiment, the pharmaceutical composition is used to treat autoimmune diseases, including multiple sclerosis (MS), myasthenia gravis (MG), chronic spontaneous urticaria, neuromyelitis optica, systemic lupus erythematosus (SLE), or rheumatoid arthritis (RA).

[0030] In another preferred embodiment, the pharmaceutical composition is used to treat cancers including (but not limited to): lymphoma, leukemia, non-small cell lung cancer, uterine cancer, colorectal cancer, brain cancer, head cancer, neck cancer, bladder cancer, prostate cancer, breast cancer, kidney cancer, liver cancer, gastric cancer, or pancreatic cancer.

[0031] In the fifth aspect of the present invention, a treatment method is provided, which comprises the steps of administering the compound described in the first aspect of the present invention, its optical isomer or mixture thereof, its crystalline form, its salt, its hydrate or solvate, or administering the pharmaceutical composition described in the third aspect of the present invention to a subject in need of treatment, thereby inhibiting BTK.

[0032] 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.

[0033] 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%.

[0034] In another preferred embodiment, the compound of formula I contains at least 1 or 3 deuterium atoms, more preferably 5 or 8 deuterium atoms.

[0035] As used herein, the term "compound of the present invention" refers to a compound represented by Formula I. The term also includes optical isomers of the compound of Formula I or mixtures thereof, crystal forms, salts thereof, hydrates thereof or solvates thereof.

[0036] 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.

[0037] The positive and progressive effects of the present invention are:

[0038] (1) The compounds of the present invention have good selective BTK inhibitory effects and can be effectively used for treating diseases associated with BTK.

[0039] (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.

[0040] (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 drugability.

[0041] Specific implementation methods

[0042] The present invention will be further described below in conjunction with specific examples. It should be understood that 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.

[0043] 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.

[0044] 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 using the corresponding deuterated starting compounds as raw materials and in the same way.

[0045] Example 1: Synthesis of Compound T109

[0046] Synthetic route

[0047]

[0048] Step 1: Synthesis of compound T003

[0049]

[0050] 2,4-dichloro-3-nitropyridine (41.5mmol), tert-butyl (R)-3-aminopiperidine-1-carboxylate (41.4mmol) and TEA (62.2mmol) were added to N,N-dimethylformamide (50ml), and 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 with anhydrous sodium sulfate and concentrated. Purification was performed on a silica gel column with ethyl acetate / petroleum ether (1:1) as the eluent, and the solvent was removed to obtain compound T003 as a yellow oil.

[0051] Step 2: Synthesis of compound T005

[0052]

[0053] Compound T003 (22.4 mmol), bis[(4-methoxyphenyl)methyl]amine (compound T004) (22.4 mmol) and TEA (29.5 mmol) were added to isopropanol (100 ml), and the resulting reaction mixture was stirred at 95° C. overnight, cooled and concentrated in vacuo to obtain compound T005 as a yellow oil.

[0054] Step 3: Synthesis of compound T006

[0055]

[0056] Compound T005 (17.3 mmol) and Fe (173.1 mmol) were added to a solution of AcOH / MeOH (1:1, 100 mL), and the reaction mixture was stirred at 25°C overnight, and then concentrated in vacuo, and the pH value 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 obtain compound T006 as a yellow oil.

[0057] Step 4: Synthesis of compound T007

[0058]

[0059] Compound T006 (20.2 mmol) and CDI (30.1 mmol) were added to CH3CN (100 ml), and the reaction mixture was stirred at 80°C overnight. The reaction mixture was cooled and concentrated. Purification was performed on a silica gel column with ethyl acetate / petroleum ether (1:5) as the eluent, and the solvent was removed to obtain compound T007.

[0060] Step 5: Synthesis of compound T009

[0061]

[0062] Compound T007 (17.5 mmol), penta-deuterated (4-phenoxyphenyl) boronic acid (compound T008) (35.1 mmol), TEMPO (19.5 mmol) and TEA (69.5 mmol), Cu (OAc) 2 (8.9 mmol) were added to dichloromethane (100 ml). The reaction mixture was stirred overnight at 25 ° C under an oxygen atmosphere. Penta-deuterated (4-phenoxyphenyl) boronic acid (compound T008) (35.1 mmol) was added, and the reaction mixture was reacted overnight at 25 ° C. Purification was performed on a silica gel column, and the eluent was ethyl acetate / petroleum ether (1:3). The solvent was removed to obtain compound T009.

[0063] Step 6: Synthesis of compound T010

[0064]

[0065] Compound T009 (6.1 mmol) and trifluoroacetic acid (80 ml) were added to dichloromethane (80 ml). The resulting reaction mixture was stirred at 50°C for 5 hours, concentrated in vacuo, and the pH value of the residual solution was adjusted to 9 with sodium bicarbonate. The resulting solution was extracted with dichloromethane, and the organic layers were combined and dried over anhydrous sodium sulfate. The mixture was purified by silica gel column with dichloromethane / methanol (30:1) as the eluent, and the solvent was removed to obtain compound T010.

[0066] Step 7: Synthesis of compound T109

[0067]

[0068] Compound T010 (150 mg, 0.37 mmol, 1.00 equivalent) and TEA (113 mg, 1.12 mmol, 3.00 equivalent) were added to DCM-CH3OH (1:1, 6 ml). Trideuterated prop-2-enoyl chloride (compound T011) (40.1 mg, 0.44 mmol, 1.20 equivalent) was then added dropwise at 0°C with stirring within 5 minutes. The resulting solution was stirred at 0°C for 2 hours and concentrated in vacuo. The residue was purified by silica gel column, eluent dichloromethane / methanol (30:1), and the solvent was removed to obtain a crude product. The crude product was prepared by C 18 The product was purified by chromatography column with a mobile phase of 0.05% TFA and ACN in gradient elution to obtain compound T109.

[0069] The H NMR spectrum of compound T109 is: 1H-NMR(DMSO-d6)δ7.8(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).

[0070] Example 2: Synthesis of Compound T108

[0071] For the synthesis of compound T108, the compound T008 in step 5 of "Example 1: Synthesis of compound T109" was replaced with compound T201. The remaining steps were the same as those in "Example 1: Synthesis of compound T109" to obtain compound T108. The H NMR spectrum of compound T108 is: 1 H-NMR(DMSO-d6)δ7.8(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).

[0072]

[0073] Example 3: Synthesis of Compound T107

[0074] For the synthesis of compound T107, the compound T008 in step 5 of "Example 1: Synthesis of compound T109" was replaced with compound T202. The remaining steps were performed in the same manner as in "Example 1: Synthesis of compound T109" to obtain compound T107. The H NMR spectrum of compound T107 is: 1 H-NMR(DMSO-d6)δ7.8(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).

[0075]

[0076] Example 4: Synthesis of Compound T106

[0077] For the synthesis of compound T106, the compound T008 in step 5 of "Example 1: Synthesis of compound T109" was replaced with compound T203. The remaining steps were performed in the same manner as in "Example 1: Synthesis of compound T109" to obtain compound T106. The H NMR spectrum of compound T106 is:1 H-NMR(DMSO-d6)δ7.8(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).

[0078]

[0079] Example 5: Synthesis of Compound T105

[0080] For the synthesis of compound T105, the compound T008 in step 5 of "Example 1: Synthesis of compound T109" was replaced with compound T204. The remaining steps were performed in the same manner as in "Example 1: Synthesis of compound T109" to obtain compound T105. The H NMR spectrum of compound T105 is: 1 H-NMR(DMSO-d6)δ7.8(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).

[0081]

[0082] Example 6: Synthesis of Compound T104

[0083] For the synthesis of compound T104, the compound T011 in step 7 of "Example 1: Synthesis of compound T109" was replaced with compound T205. The remaining steps were performed in the same manner as in "Example 1: Synthesis of compound T109" to obtain compound T104. The H NMR spectrum of compound T104 is: 1 H-NMR(DMSO-d6)δ7.8(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).

[0084]

[0085] Example 7: Synthesis of Compound T103

[0086] For the synthesis of compound T103, the compound T008 in step 5 of "Example 6: Synthesis of compound T104" was replaced with compound T201. The remaining steps were the same as those in "Example 6: Synthesis of compound T104" to obtain compound T103. The H NMR spectrum of compound T103 is: 1 H-NMR(DMSO-d6)δ7.8(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)

[0087]

[0088] Example 8: Synthesis of Compound T102

[0089] For the synthesis of compound T102, the compound T008 in step 5 of "Example 6: Synthesis of compound T104" was replaced with compound T202. The remaining steps were the same as those in "Example 6: Synthesis of compound T104" to obtain compound T102. The H NMR spectrum of compound T102 was: 1 H-NMR(DMSO-d6)δ7.8(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).

[0090]

[0091] Example 9: Synthesis of Compound T101

[0092] For the synthesis of compound T101, the compound T008 in step 5 of "Example 6: Synthesis of compound T104" was replaced with compound T203. The remaining steps were the same as those in "Example 6: Synthesis of compound T104" to obtain compound T101. The H NMR spectrum of compound T101 is: 1H-NMR(DMSO-d6)δ7.8(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).

[0093]

[0094] Example 10: Synthesis of Compound T113

[0095] For the synthesis of compound T113, the compound T008 in step 5 of "Example 6: Synthesis of compound T104" was replaced with compound T206. The remaining steps were performed in the same manner as in "Example 6: Synthesis of compound T104" to obtain compound T113. The H NMR spectrum of compound T113 is: 1 H-NMR(DMSO-d6)δ7.8(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).

[0096]

[0097] Example 11: Synthesis of Compound T114

[0098] For the synthesis of compound T114, the compound T008 in step 5 of "Example 6: Synthesis of compound T104" was replaced with compound T207. The remaining steps were the same as those in "Example 6: Synthesis of compound T104" to obtain compound T114. The H NMR spectrum of compound T114 is: 1 H-NMR(DMSO-d6)δ7.8(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).

[0099]

[0100] Example 12: Synthesis of Compound T115

[0101] For the synthesis of compound T115, the compound T008 in step 5 of "Example 6: Synthesis of compound T104" was replaced with compound T208. The remaining steps were performed in the same manner as in "Example 6: Synthesis of compound T104" to obtain compound T115. The H NMR spectrum of compound T115 is: 1 H-NMR(DMSO-d6)δ7.8(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).

[0102]

[0103] Example 13: Synthesis of Compound T112

[0104] For the synthesis of compound T112, the compound T008 in step 5 of "Example 1: Synthesis of compound T109" was replaced with compound T208. The remaining steps were performed in the same manner as in "Example 1: Synthesis of compound T109" to obtain compound T112. The H NMR spectrum of compound T112 is: 1 H-NMR(DMSO-d6)δ7.8(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).

[0105]

[0106] Example 14: Synthesis of Compound T111

[0107] For the synthesis of compound T111, the compound T008 in step 5 of "Example 1: Synthesis of compound T109" was replaced with compound T207. The remaining steps were the same as those in "Example 1: Synthesis of compound T109" to obtain compound T111. The H NMR spectrum of compound T111 is: 1 H-NMR(DMSO-d6)δ7.8(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).

[0108]

[0109] Example 15: Synthesis of Compound T110

[0110] For the synthesis of compound T110, the compound T008 in step 5 of "Example 1: Synthesis of compound T109" was replaced with compound T206. The remaining steps were performed in the same manner as in "Example 1: Synthesis of compound T109" to obtain compound T110. The H NMR spectrum of compound T110 is: 1 H-NMR(DMSO-d6)δ7.8(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).

[0111]

[0112] Example 16: Synthesis of Compound T008

[0113]

[0114] Step 8: Synthesis of compound T032

[0115] Take phenol-d5 (compound T030) (9.4 g), dissolve it in anhydrous tetrahydrofuran (100 ml), stir, 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 hours, filter the reactant, spin dry the filtrate, add dichloromethane to dissolve, pass through a column, and elute with petroleum ether: ethyl acetate (1:5) to obtain compound T032.

[0116] Step 9: Synthesis of compound T008

[0117] Compound T032 (4.5 g) was dissolved in dry THF (100 ml), and the mixture was reacted at -78 °C for 30 min under N2 protection. n-Butyl lithium (1.7 g) was slowly added dropwise, and the mixture was kept at -78 °C for 3 h after the addition was completed. Triisopropyl borate (3.8 g) was then slowly added dropwise, and the mixture was kept at -78 °C for 2 h after the addition was completed. The mixture was slowly warmed to room temperature and reacted 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 T008.

[0118] Example 17: Synthesis of Compound T202

[0119] The synthesis of compound T202 was carried out according to "Example 16: Synthesis of compound T008", except that in step 8, compound T030 was replaced by compound T033, and the remaining steps were carried out in the same manner as "Example 16: Synthesis of compound T008" to obtain compound T202.

[0120]

[0121] Example 18: Synthesis of Compound T201

[0122] The synthesis of compound T201 was carried out according to "Example 16: Synthesis of compound T008", except that in step 8, compound T030 was replaced by compound T034, and the remaining steps were carried out in the same manner as "Example 16: Synthesis of compound T008" to obtain compound T201.

[0123]

[0124] Example 19: Synthesis of Compound T203

[0125] The synthesis of compound T203 was carried out according to "Example 16: Synthesis of compound T008", except that in step 8, compound T030 was replaced by compound T035, and the remaining steps were carried out in the same manner as "Example 16: Synthesis of compound T008" to obtain compound T203.

[0126]

[0127] Example 20: Synthesis of Compound T206

[0128] The synthesis of compound T206 was carried out according to "Example 16: Synthesis of compound T008", except that in step 8, compound T030 was replaced by compound T036, and the remaining steps were carried out in the same manner as "Example 16: Synthesis of compound T008" to obtain compound T206.

[0129]

[0130] Example 21: Synthesis of Compound T207

[0131] The synthesis of compound T207 was carried out according to "Example 16: Synthesis of compound T008", except that in step 8, compound T030 was replaced by compound T037, and the remaining steps were carried out in the same manner as "Example 16: Synthesis of compound T008" to obtain compound T207.

[0132]

[0133] Example 22: Synthesis of Compound T208

[0134] The synthesis of compound T208 was carried out according to "Example 16: Synthesis of compound T008", except that in step 8, compound T030 was replaced by compound T038, and the remaining steps were carried out in the same manner as "Example 16: Synthesis of compound T008" to obtain compound T208.

[0135]

[0136] Example 23: Pharmacokinetic Evaluation in Rats

[0137] 48 male Sprague-Dawley rats, 7-8 weeks old, 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), 6 rats in each group. Tolebrutinib, compound T101, compound T104, compound T105, compound T106, compound T107, compound T109 and compound T112 were given a single oral gavage at a dose of 6 mg / kg according to the grouping, and their pharmacokinetic differences were compared.

[0138] Rats were fed with standard feed and 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, 10 hours and 24 hours after administration.

[0139] After the blood sample is collected, it is placed in a centrifuge tube coated with sodium heparin solution. Immediately and gently invert the tube at least 5 times to ensure sufficient mixing and then place it on ice. The blood sample is centrifuged at 5000 rpm 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, mark the sample number and blood collection time point. The plasma is stored in a -80°C refrigerator before LC-MS / MS analysis.

[0140] The experimental results show that compared with Tolebrutinib, the elimination half-life of compound T101 is T 1 / 2 and / or area under the curve AUC and / or maximum plasma concentration C max Compared with Tolebrutinib, the elimination half-life of compound T104 was increased by more than 40%. 1 / 2 and / or area under the curve AUC and / or maximum plasma concentration C max Compared with Tolebrutinib, the elimination half-life of compound T105 was increased by more than 40%. 1 / 2and / or area under the curve AUC and / or maximum plasma concentration C max The elimination half-life of compound T106 increased by more than 50% compared with Tolebrutinib. 1 / 2 and / or area under the curve AUC and / or maximum plasma concentration C max Compared with Tolebrutinib, the elimination half-life of compound T107 was T 1 / 2 and / or area under the curve AUC and / or maximum plasma concentration C max Compared with Tolebrutinib, the elimination half-life of compound T109 was T 1 / 2 and / or area under the curve AUC and / or maximum plasma concentration C max Increased by more than 50%; Compared with Tolebrutinib, the elimination half-life of compound T112 was T 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 compound T101, compound T104, compound T105, compound T106, compound T107, compound T109 and compound T112 of the present invention have better pharmacokinetic properties in animals, indicating better pharmacodynamics and therapeutic effects.

[0142] Example 24: 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 light signals were read using an EnVision microplate reader (Perkin Elmer). The detection buffer included 40mM Tris-HCl (pH7.5), 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 ) / (RLU max -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)]]> T101 1.6 T104 1.7 T103 1.9 T108 1.8 T105 1.1 T106 1.4 T109 0.92 T112 2.1 Tolebrutinib 2.3

[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 25: 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 + Tolebrutinib group. The model + example compound group was intragastrically administered with the example compound once a day at a dose (50 mg / kg); the model + Tolebrutinib group was intragastrically administered with Tolebrutinib once a day at a dose (50 mg / kg), for 8-16 weeks respectively, 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 mice in the model group, model + example compound group and model + Tolebrutinib group were intraperitoneally injected with 250 mg / kg of APAP saline solution once. 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 + 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 significant effect on lipid peroxidation caused by APAP, while Tolebrutinib (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 Tolebrutinib 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+Tolebrutinib group A++ B++

[0166] Marking (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.

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

[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 deuterated aminopyridine derivative represented by formula I, its optical isomers or mixtures thereof, its crystal forms, its salts, its hydrates or solvates: 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.

2. The compound according to claim 1, characterized in that The compound is selected from the group consisting of:

3. A pharmaceutical composition, characterized in that Comprising the compound according to claim 1 or 2 or a pharmaceutically acceptable salt, isomer, metabolite, prodrug, solvate or hydrate thereof and a pharmaceutically acceptable carrier or excipient.

4. Use of the compound according to claim 1 or 2 or a pharmaceutically acceptable salt, isomer, metabolite, prodrug, solvate or hydrate thereof in the preparation of a BTK inhibitor.

5. Use of the compound according to claim 1 or 2 or a pharmaceutically acceptable salt, isomer, metabolite, prodrug, solvate or hydrate thereof in the preparation of a medicament for treating and / or preventing a disease associated with BTK.

6. The use according to claim 5, characterized in that: The BTK-related disease is an allergic disorder, an autoimmune disease, an inflammatory disease, a thromboembolic disease or a cancer.

7. The use according to claim 6, characterized in that: The autoimmune disease is selected from multiple sclerosis, systemic lupus erythematosus, chronic spontaneous urticaria, neuromyelitis optica or rheumatoid arthritis.

8. The use according to claim 6, characterized in that: The cancer is selected from lymphoma, leukemia, non-small cell lung cancer, uterine cancer, colorectal cancer, brain cancer, head cancer, neck cancer, bladder cancer, prostate cancer, breast cancer, kidney cancer, liver cancer, stomach cancer or pancreatic cancer.

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