Sulfimide substituted indazole IRAK4 kinase inhibitor, preparation method and application
1. A sulfimide-substituted indazole compound, or isomers and pharmaceutically acceptable salts thereof, wherein the structure of the compound is shown as formula I:
wherein the content of the first and second substances,
a is selected from
R1Selected from hydrogen, cyano, halogen, C1-C6Alkyl radical, C1-C6Alkyl hydroxy, C1-C6Alkoxy or C3-C8A cycloalkyl group;
or R1Is morpholinyl or tetrahydropyrrolyl, or is substituted by one or more hydroxy groups or C1-C6Alkyl-substituted morpholinyl or tetrahydropyrrolyl;
R2selected from hydrogen, C1-C6Alkyl radical, C3-C8A cycloalkyl group; said C is1-C6Alkyl radical, C3-C8Cycloalkyl groups may be substituted with one or more halogen;
ar is selected from aryl or heteroaryl, optionally substituted with one or more R5Substituted by groups;
R5selected from hydrogen, cyano, halogen, C1-C6Alkyl radical, C1-C6Alkoxy or C3-C8A cycloalkyl group;
R6selected from hydrogen, halogen or C1-C6An alkyl group.
2. The sulfenimide-substituted indazole of claim 1, or isomers, pharmaceutically acceptable salts thereof, wherein:
a is selected from
R1Selected from hydrogen, cyano, halogen, C1-C6Alkyl radical, C1-C6Alkoxy or C3-C8A cycloalkyl group;
or R1Comprises the following steps:
R2selected from hydrogen, C1-C6Alkyl radical, C3-C8A cycloalkyl group;
R3and R4Selected from hydrogen or C1-C6An alkyl group;
ar is selected from the group consisting of5A group-substituted benzene, pyridine, pyrimidine, quinoline, quinazoline, thiophene, thiazole, or oxazole ring;
R5selected from hydrogen, cyano, halogen, C1-C6Alkyl radical, C1-C6Alkoxy or C3-C8A cycloalkyl group;
R6selected from hydrogen or C1-C6An alkyl group.
3. The sulfenimide-substituted indazole of claim 1, or isomers, pharmaceutically acceptable salts thereof, wherein:
a is selected from
R1Selected from hydrogen, cyano, halogen, C1-C3Alkyl radical, C1-C3Alkoxy or C3-C8A cycloalkyl group;
or R1Comprises the following steps:
R2selected from hydrogen, C1-C3Alkyl radical, C3-C8A cycloalkyl group;
R3and R4Always have the same definition and are each selected from hydrogen or C1-C3An alkyl group;
ar is selected from the group consisting of one, two or three R5Radical (I)A substituted benzene, pyridine, pyrimidine, quinoline, quinazoline, thiophene, thiazole, or oxazole ring;
R5selected from hydrogen, cyano, halogen or C1-C3An alkyl group;
R6selected from hydrogen or C1-C3An alkyl group.
4. The sulfenimide-substituted indazole, or isomers thereof, and pharmaceutically acceptable salts thereof, according to claim 1, wherein said sulfenimide-substituted indazole has the structure shown in the following table:
TABLE 1
5. A process for the preparation of a sulfenimide-substituted indazole according to any one of claims 1-4, comprising the steps of:
(1) carrying out condensation reaction on the compound IA and the compound IB to prepare IC;
(2) reacting the compound IC with a side chain ID to obtain a final product I; the reaction formula of the method is as follows:
6. use of a sulfinimide-substituted indazole compound or isomer, pharmaceutically acceptable salt thereof according to any one of claims 1-4, for the preparation of a medicament for the prevention or treatment of IRAK 4-related diseases.
7. The use of claim 6, wherein the disease is selected from the group consisting of autoimmune disease; inflammatory diseases; pain disorders; respiratory, airway and lung diseases; lung inflammation and injury; pulmonary hypertension; gastrointestinal disorders; allergic diseases; infectious diseases; trauma and tissue injury disorders; fibrotic diseases; eye diseases; joint, muscle and bone diseases; skin diseases; kidney disease; diseases of the hematopoietic system; liver disease; oral diseases; metabolic diseases, heart diseases; vascular disease; a neuroinflammatory disorder; neurodegenerative diseases; sepsis; a genetic disease; and cancer.
8. The use according to claim 7, wherein the autoimmune and inflammatory diseases are selected from systemic lupus erythematosus, lupus nephritis, arthritis, psoriasis, colitis, Crohn's disease, atopic dermatitis, liver fibrosis, myelofibrosis, thrombocythemia, polycythemia, senile dementia, gout, protein-related periodic syndrome, chronic or acute kidney injury, chronic obstructive pulmonary disease, asthma, bronchospasm, or graft-versus-host disease.
9. The use according to claim 7, wherein the cancer is selected from breast cancer, small cell lung cancer, non-small cell lung cancer, bronchoalveolar carcinoma, prostate cancer, bile duct cancer, bone cancer, bladder cancer, head and neck cancer, kidney cancer, liver cancer, cancer of gastrointestinal tissues, cancer of esophagus, ovary cancer, pancreas cancer, skin cancer, testicular cancer, thyroid cancer, uterine cancer, cervical and vaginal cancer, leukemia, multiple myeloma or lymphoma.
10. A composition comprising a therapeutically effective amount of the sulfenimide substituted indazole compound of any one of claims 1-4, or isomers, pharmaceutically acceptable salts thereof, and a pharmaceutically acceptable carrier.
Background
Interleukin-1 receptor-associated kinase 4(IRAK-4) is one of the members of the IRAK family of intracellular serine-threonine kinases. Other members of the kinase family also include IRAK-1, IRAK-2 and IRAK-M. IRAK-M is expressed only in monocytes and macrophages, and expression of IRAK-1, IRAK-2 and IRAK4 is ubiquitous. IRAK4 is composed mainly of a conserved Death Domain (DD) at the N-terminus, a hinge region, and a central Kinase Domain (KD) at the C-terminus. The DD region is the region where IRAK4 binds to the initial response gene 88(MyD88) of the adaptor myeloid differentiation factor. The KD region consists of 12 subregions, with typical serine-threonine kinase domain characteristics. The primary function of IRAK4 is to phosphorylate its substrate via the KD region, thereby activating downstream signaling molecules. IRAK4 is a key factor downstream of the inflammatory signaling pathway mediated by interleukin-1 receptor (IL-1R)/Toll-like receptor (TLR), and plays a critical role in the immune system (Sims JE, et. natrevmjnol, 2010,10(2):89-102). Upon binding of the ligand to interleukin-1 receptor (IL-1R) or Toll-like receptor (TLR), IRAK4 is able to mediate signaling, activating expression of downstream inflammatory factors. TLRs can receive ligand signals from the body upon interaction with microorganisms or stimulation by endogenous substances, as well as first wave inflammatory signals and innate immune response signals elicited by these stimuli. TLRs play a very important role in many diseases, including infections and auto-inflammatory diseases, as well as many other diseases in humans. Like cancer necrosis factor-alpha (TNF-alpha) and other major cytokines, interleukin-1 (IL-1) is a key factor in the inflammation-mediated pathway, capable of transmitting and amplifying signals. Because TLR, IL-1R and other cytokine receptor mediated signal pathways have mutual cross-linking effect, IRAK4, a key signal factor in TLR and IL-1R inflammation pathway plays a significant role in systemic inflammation reaction, and can be used as an effective potential target spot for treating various inflammation-related diseases.
It was determined that a subset of human patients lack IRAK4 expression (Picard, C.et al, 2003, Science 299: 2076-. Deletion of IRAK4 in mice results in severe blocking of IL-1, IL-18 and all TLR-dependent responses (except TLR3) (Suzuki, N.et al, 2002, Nature 416: 750-. In contrast, deletions of IRAK1(Thomas, J.A.et al, 1999, J.Immunol.163: 978-. Moreover, IRAK4 is the only family member in the IRAK family whose kinase activity has been shown to be essential for initiating signaling. The replacement of wild-type IRAK4 in the mouse genome with a mutant with inactivated kinase activity (KDKI) blocked all signals transmitted by MyD 88-dependent receptors, including IL-1, IL-18 and all T L Rs (except T L R3) (Koziczak-Holbro, M.et al, 2007, J.biol.chem.282: 13552. 13560; Kawagoe, T.et al, 2007, J.exp.Med.204: 1013. 1024).
Mice harboring mutant (KDKI) with inactivated IRAK4 kinase activity show a large reduction in disease severity in disease models of multiple sclerosis (Staschke, K.A. et al, 2009, J.Immunol.183: 568-577), rheumatoid Arthritis (Koziczak-Holbro, M.et al, 2009, Arthritis Rheum.60: 1661-1671), atherosclerosis (Kim, T.W.et al, 2011, J.Immunol.186: 2871-2880) and myocardial infarction (Maekawa, Y.et al, 2009, Circulation 120: 1401-1414) compared to wild-type mice. As described above, IRAK4 inhibitors block all MyD88 dependent signaling. MyD 88-dependent TLRs have been shown to be responsible for the following conditions: multiple sclerosis, rheumatoid arthritis, cardiovascular disease, metabolic syndrome, sepsis, systemic lupus erythematosus, inflammatory bowel disease (including crohn's disease and ulcerative colitis), autoimmune uveitis, asthma, allergy, type I diabetes, and rejection after organ transplantation (Keogh, b.et al, 2011, Trends pharmacol. sci.32: 435-. Among diffuse large B-cell lymphomas, tumor cells harboring an oncogenic MyD88 mutation have been determined to be sensitive to IRAK4 inhibition (Ngo, V.et al, 2011, Nature 470: 115-one 121). Whole genome sequencing also confirmed that the MyD88 mutation was associated with chronic lymphocytic leukemia, suggesting the possibility of use of IRAK4 inhibitors in the treatment of leukemia (Puente, X.S. et al, 2011, Nature 475: 101-.
IRAK4 inhibitors are also capable of blocking signals transmitted by IL-1 and the IL-1 family. Modulation of IL-1 has been shown to be effective in a variety of diseases, including gout, gouty arthritis, type II diabetes, autoinflammatory diseases, tumor necrosis factor receptor-related periodic syndrome, familial mediterranean fever, adult stills disease, systemic onset juvenile idiopathic arthritis, stroke, graft-versus-host disease, asymptomatic multiple myeloma, recurrent pericarditis, osteoarthritis, and emphysema, among others (Dina relo, c.a., 2011, eur.j.immunol.41: 1203-. In a mouse model of Alzheimer's disease, blocking IL-1 receptors improves cognitive deficits, reduces tauopathies and reduces oligomeric forms of beta amyloid (Kitazawa, M.et al, 2011, J.Immunol.187: 6539-. IL-1 has also been shown to be a key link in adaptive Immunity, driving the differentiation of the effector T cell subset Th17 (Chung, Y.et al, 2009, Immunity 30: 576-587). Therefore, IRAK4 inhibitors are predicted to exhibit effects in Th17 cell-related diseases including multiple sclerosis, psoriasis, inflammatory bowel disease, autoimmune uveitis, and rheumatoid arthritis, among others (Wilke, c.m. et al, 2011, Trends immunol. 32: 603-.
Pulmonary disorders such as pulmonary fibrosis, obstructive pulmonary disease (COPD), Acute Respiratory Distress Syndrome (ARDS), Acute Lung Injury (ALI), Interstitial Lung Disease (ILD), sarcoidosis, and pulmonary hypertension have also been shown to be associated with various TLR-mediated signaling pathways. The pathogenesis of pulmonary disorders may be Infection-mediated or non-Infection-mediated processes (Jeyaseelan, Chu et al, Infection and Immunity, 2005; Seki, Tasaka et al, Inflammation Research, 2010; Nadigel, Prefontaine et al, Respiratory Research, 2011; Kovach and Standard, International Immunopharmacology, 2011; Bauer, Shapiro et al, Mol Med, 2012; Deng, Yang et al, PLoS One, 2013; Freeman, Martinez et al, Respiratory Research, 2013; Dubaniewicz, A., Human Immunology, 2013). TLRs and IL-1R family members are also involved in The pathogenesis of other inflammatory disorders such as Behcet's disease, gout, lupus erythematosus, adult Steve's disease, and chronic inflammatory bowel disease such as ulcerative colitis and Crohn's disease, as well as Transplant rejection, and therefore IRAK4 inhibitors are suitable for treating such diseases (Nickerson, Christensen et al, The Journal of Immunology, 2010; Kobori, Yagi et al, J Gastroenterol, 2010; Shi, Musci et al, Immunology Reviews, 2010; Chen, Lin et al, Arthroritis research Ther, 2013; Ha O, Liu et al, Curr O Gastroenterol, 2013; Kreisel a Gold strain, transplantation, Japan, 2013; Japan, 2013, Nematology et al; Zollin & 2013, Zollin & 2013, Zollin et al; Zollin & 2013, Zollin & S & gt, Zollin & 2013, and J. IRAK4 inhibitors are also suitable for prophylactic and/or therapeutic use for TLR and IL-1R family mediated disorders endometriosis and atherosclerosis (Akoum, Lawson et al, Human Reproduction, 2007; Allhorn, Boing et al, Reproduction Biology and Endocrinology, 2008; Lawson, Bourcer et al, Journal of Reproduction Immunology, 2008; Senevirat, Sivagunnathan et al, Clinica Chimica Acta, 2012; Khan, Kitajima et al, Journal of Obstetrics and Gyncology Research, 2013; Santuni, Borghese et al, man Reproduction, 2013; Sedumb, Halogf et al, molecular Research, 2013).
In addition to the disorders already mentioned, the IRAK 4-mediated TLR process is also described in the pathogenesis of ocular disorders such as retinal ischemia, keratitis, allergic conjunctivitis, keratoconjunctivitis sicca, macular degeneration and uveitis (Kaa rnarira nta a nd Sa lminen, J Mol Med (Berl), 2009; Sun and Pearlan, Investigative Ophthalmology & Visual Science, 2009; Redfern and McDermott, Experimental Eye Research, 2010; Kezic, Taylor et al, J Leoc Biol, 2011; Chang, McClousy et al, Clinical & Experimental Ophthalmology, 2012; Guo, Gao et al, Imonol Cell Biol, Invitro, Lee, Hatteer et al, scientific Research, Qizhi & Ophthalmology et al, Vilmology 2012; Zymology & Ouio et al).
The prior art discloses a number of IRAK4 inhibitors (see, e.g., Annual Reports in Medicinal Chemistry (2014), 49, 117-.
IRAK4 inhibitors of the indazole backbone have been extensively studied and have also been shown to be one of the effective structural classes of IRAK4 inhibitors. Wherein BAY-1834845 and BAY-1830839 are both in phase I clinical study stage. The prior literature also discloses a series of IRAK4 inhibitors of indazole structure, such as: WO2007091107a1, WO2011153588a1, WO2013106254a1, WO2015091426a1, WO2015193846a1, WO201510466a1, WO2016083433a1, WO2016174183a1, WO2017009798a1, WO2017108744a1, WO2017157792a1, WO2017207385a1, WO2017207386a1, WO2017148902a1, WO2017207481a1, WO2018060174a1, WO2018178947a1, CN110835332A, WO2019089580a1, WO2020035019a1, WO2020048471a 1. Similar non-indazole structural IRAK4 inhibitors have also been reported in the literature: WO2020035020A1, CN109890829A, CN110770229A, CN110785418A, CN111094292A, CN110835338A, WO2017207340A1 and WO2018234345A 1.
Although there are published reports on early clinical application of IRAK4 inhibitor class, no drug targeting the target is currently on the market, and only PF-06650833, BAY-1834845, BAY-1830839, R835 and CA-4948 enter clinical stage.
BAY-1830839, BAY-1834845 in phase I of the clinic are indazole IRAK4 inhibitors.
Patent document CN111362920A discloses a class of alkyl sulfimide indazoles, which are represented by the following compounds:
the applicant discovers that the indazole type IRAK4 inhibitor has certain defects through systematic scientific experimental research, and has a larger promotion space. The main defects are as follows: the problems of low inhibitory activity of IRAK4, animal safety risk, low pharmacokinetics and bioavailability and the like.
The compounds disclosed in the prior art, as well as the experimental drugs, are still unsatisfactory in terms of efficacy, safety, pharmacokinetics, etc., and there is still a need to continue the research and development of new IRAK4 inhibitors to meet the ever-increasing medical and health needs of people.
Disclosure of Invention
The technical problem to be solved by the invention is as follows: provided is a sulfimide-substituted indazole-type IRAK4 kinase inhibitor having excellent IRAK4 inhibitory activity, animal safety and pharmacokinetic parameters.
The technical scheme for solving the technical problems is as follows:
a sulfimide-substituted indazole compound, or isomers and pharmaceutically acceptable salts thereof, wherein the structure of the compound is shown as formula I:
wherein the content of the first and second substances,
a is selected from
R1Selected from hydrogen, cyano, halogen, C1-C6Alkyl radical, C1-C6Alkyl hydroxy, C1-C6Alkoxy or C3-C8A cycloalkyl group;
or R1Is morpholinyl or tetrahydropyrrolyl, or is substituted by one or more hydroxy groups or C1-C6Alkyl-substituted morpholinyl or tetrahydropyrrolyl;
R2selected from hydrogen, C1-C6Alkyl radical, C3-C8A cycloalkyl group; said C is1-C6Alkyl radical, C3-C8Cycloalkyl groups may be substituted with one or more halogen;
ar is selected from aryl or heteroaryl, optionally substituted with one or more R5Substituted by groups;
R5selected from hydrogen, cyano, halogen, C1-C6Alkyl radical, C1-C6Alkoxy radical, C3-C8A cycloalkyl group;
R6selected from hydrogen, halogen or C1-C6An alkyl group.
Preferably, the first and second liquid crystal materials are,
a is selected from
R1Selected from hydrogen, cyano, halogen, C1-C6Alkyl radical, C1-C6Alkoxy or C3-C8A cycloalkyl group;
or R1Comprises the following steps:
R2selected from hydrogen, C1-C6Alkyl radical, C3-C8A cycloalkyl group;
R3and R4Always have the same definition and are each selected from hydrogen or C1-C6An alkyl group;
ar is selected from the group consisting of5A group-substituted benzene, pyridine, pyrimidine, quinoline, quinazoline, thiophene, thiazole, or oxazole ring; in addition to the aromatic rings listed above, Ar may also be selected from other common aromatic ring structures.
R5Selected from hydrogen, cyano, halogen, C1-C6Alkyl radical, C1-C6Alkoxy radical, C3-C8A cycloalkyl group;
R6selected from hydrogen or C1-C6An alkyl group.
It is further preferred that the first and second liquid crystal compositions,
a is selected from
R1Selected from hydrogen, cyano, halogen, C1-C3Alkyl radical, C1-C3Alkoxy or C3-C8A cycloalkyl group;
or R1Comprises the following steps:
R2selected from hydrogen, C1-C3Alkyl radical, C3-C8A cycloalkyl group;
R3and R4Always have the same definition and are each selected from hydrogen or C1-C3An alkyl group;
ar is selected from the group consisting of one, two or three R5A group-substituted benzene, pyridine, pyrimidine, quinoline, quinazoline, thiophene, thiazole, or oxazole ring;
R5selected from hydrogen, cyano, halogen or C1-C3An alkyl group;
R6selected from hydrogen or C1-C3An alkyl group.
As described in this application "C1-C3The "alkyl group" of (a) means a methyl group, an ethyl group, an n-propyl group or an isopropyl group; said "C1-C3The "alkoxy group" of (a) means a methoxy group, an ethoxy group, a n-propoxy group, an isopropoxy group; the halogen refers to F, Cl, Br and I; said "C3-C8Cycloalkyl "means cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl or cyclooctyl.
Typical compounds of the present invention include, but are not limited to, the following compounds of table 1:
TABLE 2
The second purpose of the invention is to provide a synthesis method of the compound, which comprises the following steps:
(1) carrying out condensation reaction on the compound IA and the compound IB to prepare IC;
(2) the compound IC reacts with the side chain ID to prepare the final product I.
The definition of each group in the reaction step is as described above.
In a third aspect the present invention provides the use of a compound of formula (I), or an isomer, a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for inhibiting a protein kinase.
In some embodiments, the protein kinase is an IRAK family kinase, particularly an IRAK4 kinase.
In another aspect of the invention there is provided the use of a compound of formula (I), or an isomer, pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the treatment of a disease caused by a protein kinase.
In some embodiments, the compounds or compositions of the present invention may be used to treat autoimmune diseases caused by IRAK family kinases, particularly IRAK4 kinase; inflammatory diseases; pain disorders; respiratory, airway and lung diseases; lung inflammation and injury; pulmonary hypertension; gastrointestinal disorders; allergic diseases; infectious diseases; trauma and tissue injury disorders; fibrotic diseases; eye diseases; joint, muscle and bone diseases; skin diseases; kidney disease; diseases of the hematopoietic system; liver disease; oral diseases; metabolic diseases, heart diseases; vascular disease; a neuroinflammatory disorder; neurodegenerative diseases; sepsis; a genetic disease.
In some embodiments, the autoimmune and inflammatory diseases described herein are selected from: systemic Lupus Erythematosus (SLE), lupus nephritis, arthritis, psoriasis, colitis, crohn's disease, atopic dermatitis, liver fibrosis, myelofibrosis, thrombocythemia, polycythemia, senile dementia, gout, protein-associated periodic syndrome (CAPS), chronic kidney disease or acute kidney injury, Chronic Obstructive Pulmonary Disease (COPD), asthma, bronchospasm, and graft-versus-host disease.
In some embodiments, the compounds or compositions of the present invention may be used to treat diseases of abnormal cellular proliferation, particularly cancer, caused by IRAK family kinases, particularly IRAK4 kinases.
In some embodiments, the cancer of the invention includes breast cancer, small cell lung cancer, non-small cell lung cancer, bronchoalveolar carcinoma, prostate cancer, bile duct cancer, bone cancer, bladder cancer, head and neck cancer, kidney cancer, liver cancer, cancer of the gastrointestinal tissue, esophageal cancer, ovarian cancer, pancreatic cancer, skin cancer, testicular cancer, thyroid cancer, uterine cancer, cervical and vaginal cancer, leukemia, multiple myeloma, and lymphoma.
The derivative can be formed into a composition for treating related cancers and other diseases by oral administration, injection and the like in the process of treating diseases. For oral administration, it can be prepared into conventional solid preparations such as tablet, powder or capsule; for injection, it can be prepared into injection.
In a third aspect of the invention, there is provided a composition comprising a therapeutically effective amount of the novel aromatic (hetero) sulfinimide-substituted indazole compound described above, or isomers, pharmaceutically acceptable salts, and a pharmaceutically acceptable carrier.
Pharmaceutically acceptable salts, for example, salts with inorganic acids, salts with organic acids. Non-limiting examples of salts with inorganic acids include, but are not limited to, salts with hydrochloric acid, hydrobromic acid, nitric acid, sulfuric acid, phosphoric acid, and the like. Non-limiting examples of salts with organic acids include, but are not limited to, salts with formic acid, acetic acid, trifluoroacetic acid, fumaric acid, oxalic acid, malic acid, maleic acid, tartaric acid, citric acid, succinic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, and the like.
The mentioned carriers refer to the carriers conventional in the pharmaceutical field, such as: diluents, excipients such as water, etc.; binders such as cellulose derivatives, gelatin, polyvinylpyrrolidone, etc.; fillers such as starch and the like; disintegrating agents such as calcium carbonate, sodium bicarbonate; in addition, other adjuvants such as flavoring agents and sweeteners may also be added to the composition.
The various dosage forms of the composition of the present invention can be prepared by conventional methods in the medical field, wherein the content of the active ingredient is 0.1-99.5% (by weight).
The administration amount of the present invention may vary depending on the route of administration, age, body weight of the patient, type and severity of the disease to be treated, etc., and the daily dose thereof is 0.001 to 30mg/kg body weight (oral administration) or 0.005 to 30mg/kg body weight (injection).
Compared with the prior indazole IRAK4 inhibitors (such as BAY-1834845 and BAY-1830839) and MY-004-102 and MY-004-103 in a patent document CN111362920A, the compounds in the invention have the following advantages compared with the prior art except that the activity of IRAK4 is better: 1) the aryl (hetero) sulfimide structure replaces the substituent group in the prior art to obtain the unexpected effect of reducing the hERG inhibition risk; 2) the compound of the invention has initial safety superior to that of the existing indazole type IRAK4 inhibitor; 3) the animal in-vivo experimental data show that the drug parameters AUC and Cmax of the compound are obviously higher than those of the existing indazole IRAK4 inhibitor under the same dose. The compound has better PK property, so that the compound can be reasonably speculated to be applied to clinic, the effective dose is lower, and the medication safety is higher.
The Chinese naming of the compound of the invention conflicts with the structural formula, and the structural formula is taken as the standard; except for obvious errors in the formula.
Compared with the prior art, the novel aryl (hetero) sulfinyl imide substituted indazole compound and the pharmaceutically acceptable salt thereof provided by the invention have better IRAK4 inhibitory activity and better safety, and the preferred compound of the invention shows good pharmacokinetic properties and has the potential of being developed into a selective IRAK4 inhibitor.
Detailed Description
Example 1 Synthesis of I-1
The synthetic route is as follows:
the method comprises the following operation steps:
step 1:
compound IA-1(1.63g,0.01mol), IB-1(1.91g,0.01mol), dichloromethane (DCM, 30mL) were added, N-diisopropylethylamine (DIPEA, 1.94g,0.015mol), 1- (3-dimethylaminopropyl) -3-Ethylcarbodiimide (EDCI) (2.3g,0.015mol) were added, and the reaction was stirred at 30 ℃ for 12 h. The reaction mixture was extracted with water (20mL), and the organic layer was concentrated to dryness under reduced pressure and recrystallized by adding absolute ethanol (10mL) to give IC-1 as a pale yellow solid (2.67g, yield 79.5%).1H NMR(400MHz,DMSO-d6):δ＝13.10(br,1H),11.55(br,1H),8.35(d,J＝8.0Hz,1H),8.14(m,1H),7.98(d,J＝8.0Hz,1H),7.72(s,1H),7.47(s,1H),7.04(s,1H),3.92(s,3H)。LCMS:MS Calcd.:336.3,MS Found:337.2[M+1]。
Step 2:
compounds IC-1(400mg,1.2mmol), ID-1(327mg,1.32mmol), K2CO3(332mg,2.4mmol), KI (17mg,0.1mmol) were added to DMF (10mL), and N2Heating to the internal temperature of 100 ℃ under the protection of gas, and stirring for reaction for 24 hours. After the reaction, the temperature was reduced to room temperature, the solvent was concentrated under reduced pressure to dryness, and the residue was passed through a silica gel column (eluent ethyl acetate/petroleum ether: 1/2), and the product fractions were collected and concentrated to dryness to give product I-1(210mg, yield 35%).1HNMR(400MHz,DMSO-d6):δ＝11.53(br,1H),10.45(br,1H),8.36(d,J＝8.0Hz,1H),8.12(m,2H),8.02(s,1H),7.71(d,J＝8.0Hz,1H),7.51(m,5H),7.32(s,1H),4.11(t,J＝4.8Hz,2H),3.86(s,3H)，3.04(t,J＝4.8Hz,2H)。LCMS:MS Calcd.:503.5,MS Found:504.2[M+1]。
Compounds I-2 to I-36 the preparation of compound I-1 of example 1 was carried out according to the scheme given for compound I-1 and the data on the compounds are shown in Table 2.
TABLE 3
Biological assay
Test example 1, IRAK4 kinase Activity test
The inhibitory activity (IC) of compounds against IRAK4 kinase at Km ATP was determined using a Mobility Shift Assay (MSA)50) Setting 10 drug concentration gradients (initial concentration 1. mu.M, 3-fold dilution, 2 duplicate wells per concentration), adding IRAK4 kinase to the kinase-based buffer solution, transferring to the test plate, adding FAM-labeled peptide and ATP (37. mu.M), incubating at 28 ℃ for a period of time, adding 10. mu.L of stop buffer to terminate the reverse reactionThe conversion data is read by Caliper and the conversion is converted to inhibition data. Calculating IC of half inhibitory concentration by using Logit method according to inhibition rate data of each concentration50(Table 3).
TABLE 3 results of experimental kinase Activity test for Compound IRAK4
Note: the above reference and the compound of the present invention are all the same experimental values.
And (4) conclusion: the activity of the compound of the invention on IRAK4 kinase is obviously better than that of the comparative compounds BAY-1834845, BAY-1830839, MY-004-102 and MY-004-103.
Test example 2 ability of Compounds to inhibit TNF-. alpha.secretion in THP-1 cells
With the aid of this test, it may be suitable to test the ability of a compound to inhibit TNF- α (tumor necrosis factor α) secretion in THP-1 cells (human monocytic acute leukemia cell line). TNF- α is a key cytokine involved in the inflammatory process of the listed autoimmune diseases such as rheumatoid arthritis, psoriatic arthritis, ankylosing spondylitis, psoriasis, crohn's disease, ulcerative colitis, etc. In this test, TNF- α secretion was triggered by incubation with bacterial Lipopolysaccharide (LPS).
To a 96-well plate, 150. mu.L of RPMI-1640 medium solution containing 10000 THP-1 cells per well was added, followed by 25. mu.L of test compound (starting at 10. mu.M, 3-fold dilution, 9 concentrations of RPMI-1640 medium containing 4% DMSO per concentration) solution containing 8-fold final concentration, and after mixing well, incubation was performed at 37 ℃ for 30 minutes. To each test well 25. mu.L of LPS-containing RPMI-1640 medium solution (final LPS concentration 1. mu.g/mL, final DMSO concentration 0.5%) was added, mixed well and incubated at 37 ℃ for 4.5 hours. The 96-well plate is rotated at 2000rpm for 5 minutes, 50. mu.L of supernatant is taken, the TNF-alpha content in the supernatant is determined by human ELISA kit, and IC of the compound is calculated by XL-Fit50Values (table 4).
TABLE 4 Activity of the Compounds of the invention for inhibiting LPS-stimulated TNF- α secretion in THP-1 cells
Note: the above reference and the compound of the present invention are all the same experimental values.
And (4) conclusion: the compound can effectively inhibit TNF-alpha secretion stimulated by LPS in THP-1 cells, and the inhibition effect is obviously superior to that of the comparative compounds BAY-1834845, BAY-1830839, MY-004-102 and MY-004-103.
Test example 3 pharmacokinetic testing of the Compound of the invention
SD rats are used as test animals, after the compounds of BAY-1834845, BAY-1830839, MY-004-102, MY-004-103 and the preferred embodiment of the invention are administered to the rats by gastric lavage by adopting an LC/MS/MS method, the drug concentration in plasma of the rats at different times is measured, and the pharmacokinetic characteristics of the compounds in the rats are researched.
SD rat source: shanghai Slek laboratory animals Co., Ltd
The administration mode is single intragastric administration
Administration dose and concentration: 10 mg/kg; 2mg/mL
The preparation prescription is as follows: 0.5% Methelculose
Sampling points are as follows: 5min, 15min, 30min, 1h, 2h, 4h, 8h and 24h.
Preparing a standard curve and a quality control sample: appropriate amount of stock solution is diluted with 50% acetonitrile water to obtain standard working solution of 0.04, 0.10, 0.20, 0.40, 1.00, 2.00 and 4.00. mu.g/mL and quality control working solution of 0.10, 1.00 and 3.00. mu.g/mL. Respectively adding 2.50 mu L of standard curve working solution and quality control working solution into 47.5 mu L of blank rat plasma to prepare quality control samples containing standard curves with the concentrations of substances to be detected of 2.00, 5.00, 10.00, 20.00, 50.00, 100.00 and 200.00ng/mL and quality control samples with the concentrations of 5.00, 50.00 and 150.00ng/mL, respectively adding 200 mu L of acetonitrile (containing internal standard loratadine of 5ng/mL), carrying out vortex oscillation for 3min, centrifuging at 15000rpm and 4 ℃ for 15min, and taking 100 mu L of supernatant to carry out LC-MS/MS analysis. By using8.0 calculate the experimental results.
Preferred compounds of the invention have pharmacokinetic parameters as shown in table 5.
Table 5: preferred compound pharmacokinetic parameters
And (4) conclusion: the compound of the embodiment of the invention has good pharmacokinetic properties, and has obvious pharmacokinetic advantages compared with BAY-1834845, BAY-1830839, MY-004-102 and MY-004-103.
Test example 4 acute toxicity test of the Compound of the present invention
The compounds (I-1, I-2, I-5, I-7, I-10, I-16, I-21, I-24, I-26, I-29, I-31 and I-34) as well as MY-004-102 and BAY-1830839 (positive control drugs) are selected to carry out acute toxicity experiments.
(1) Experimental protocol
Firstly, the toxicity symptoms and death situations of animals after the oral administration of the compounds, such as MY-004-102, BAY-1830839 and I-1, of the invention to ICR mice are observed, and the acute toxicity is compared.
Preparing a solvent: an appropriate amount of sodium Methylcellulose (MC) is weighed, dissolved by ultrapure water to a certain volume, and prepared into 0.5% MC (w/v).
③ administration preparation: the desired test samples were weighed out separately and made up into suspensions of 12.5, 37.5, 75.0 and 100.0mg/mL with 0.5% MC solution.
Fourthly, the administration route: the test article and vehicle control group (0.5% MC) were administered orally.
Fifthly, administration frequency: single administration, with fasting overnight before dosing.
General symptom observation: the day of administration was observed 1 time about 0.5, 1, 2, 4, 6 hours after the first administration; the observation period is 2-6 days, 2 times per day, 1 time in the morning and afternoon.
Observations include, but are not limited to: general condition, behavioral activity, gait posture, eye, mouth, nose, gastrointestinal tract, dermal hair, urogenital tract.
(2) Statistical analysis
Body weight data are expressed as mean ± standard deviation and are compared between groups using the Levene's test and one-way analysis of variance, and if differences are indicated, the Dunnet test is followed.
(3) Results of the experiment
The compounds such as I-1, MY-004-102 and BAY-1830839 (positive control drugs) are selected to carry out the acute toxicity test as described above. The results are shown in Table 6.
In the MTD test, the tolerance of the animals to the drugs is considered, and the dosage is up to the time when the animals die frequently, namely the maximum tolerance.
Table 6: acute toxicity test results of compounds such as I-1 and MY-004-102 and BAY-1830839 through single oral administration
Test article
MTD(mg/kg)
MY-004-102
750
BAY-1830839
750
I-1
＞2000
I-2
＞2000
I-5
＞2000
I-7
＞2000
I-10
＞2000
I-16
＞2000
I-21
＞2000
I-24
＞2000
I-26
＞2000
I-29
＞2000
I-31
＞2000
I-34
＞2000
Note: MTD: maximum tolerated dose.
The results show that: the selected compounds such as I-1 and the like in the invention have MTD (maximum tolerance) of more than 2000mg/kg, and the acute toxicity is far lower than MY-004-102 and BAY-1830839.
Test example 5 Effect of the Compounds of the present invention on the hERG Potassium channel
Rapid activation of human delayed rectifier potassiumElectric current (I)Kr) Mainly mediated by the hERG ion channel, and is involved in repolarization of human cardiac muscle cells. The current blocked by the medicine can cause the clinical QT interval prolongation syndrome, and is easy to induce acute arrhythmia and even sudden death. The research uses a manual patch clamp method to test the effect of compounds such as I-1 on the hERG potassium current on a stable cell strain transfected with the hERG potassium channel, so as to determine whether the compounds such as I-1 have an inhibition effect on the hERG ion channel.
The final concentrations of test compounds were 5, 50 and 500. mu.M, and the final concentration of DMSO in the extracellular fluid was 0.3%. The inhibitory effect of test compounds on hERG current is shown in table 6.
Table 6: effect of compounds such as I-1 and MY-004-102, BAY-1830839 on hERG potassium channel
Under the experimental conditions, the average inhibition rate of the compounds such as I-1 of the invention on the hERG potassium channel current at 500 mu M<50% (N-3), the compounds of I-1 of the present invention have an IC50 value greater than 500. mu.M for hERG current inhibition. Comparison of the average inhibition rates of MY-004-102 and BAY-1830839 on hERG potassium channel current at 50. mu.M>50% (N-3), IC of inhibition of hERG current by MY-004-102 and BAY-183083950The value was around 50. mu.M.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.
