Amidino sulfonamide derivative and application thereof

Abstract

The invention discloses an amidino sulfonamide derivative or a pharmaceutical salt thereof, the structural general formula of the amidino sulfonamide derivative is as shown in the specification: # imgabs0. The amidino sulfonamide derivative provided by the invention not only has excellent anti-inflammatory activity, but also has lower toxicity.

Description

Technical Field

[0001] The invention belongs to the field of medical technology, and in particular relates to an amidinosulfonamide derivative and application thereof. Background Art

[0002] MCC950 is a classic, highly potent and selective NLRP3 inhibitor that can specifically inhibit NLRP3 inflammasome activation. Its chemical structure is as follows:

[0003]

[0004] In mouse models, MCC950 can effectively reduce the severity of experimental autoimmune encephalomyelitis models, and thus has the potential to be used in the treatment of inflammation-related diseases. However, in a Phase II clinical trial in patients with rheumatoid arthritis, MCC950 was found to cause elevated liver enzyme levels in serum, resulting in no new progress in clinical development.

[0005] Inflammation driven by abnormal activation of NLRP3 underlies many chronic degenerative diseases, including type II diabetes, gout, atherosclerosis, inflammatory bowel disease, and Alzheimer's disease. Therefore, inhibition of NLRP3 has become a potential therapeutic strategy for many diseases. Among them, compounds and strategies that directly inhibit the NLRP3 inflammasome by binding to the NACHT domain of NLRP3 have higher specificity. In recent years, research on NLRP3 inhibitors has made great progress, and small molecule inhibitors of various structural types have been reported. Some highly active small molecule compounds such as IFM-2427, Somalix, dapansutrile, and NT-0167 have entered clinical trial research, but no drugs have been approved for marketing yet, so it is necessary to continue to develop new structural NLRP3 inhibitors. Summary of the invention

[0006] The object of the present invention is to provide an amidinosulfonamide derivative.

[0007] Another object of the present invention is to provide a use of the amidinosulfonamide derivative in the preparation of NLRP3 inhibitors.

[0008] In order to achieve the above object, the technical solution adopted by the present invention is as follows:

[0009] In the first aspect of the present invention, there is provided an amidinosulfonamide derivative or a pharmaceutically acceptable salt thereof, the general structural formula of which is shown below:

[0010]

[0011] in:

[0012] R is selected from one of the following structures:

[0013]

[0014] R1 is selected from hydrogen, CF3, halogen (fluorine, chlorine, bromine, iodine), C1-C30 alkyl, C1-C30 alkoxy;

[0015] R2 is selected from hydrogen, CF3, halogen (fluorine, chlorine, bromine, iodine), C1-C30 alkyl, C1-C30 alkoxy;

[0016] R3 is selected from hydrogen, CF3, halogen (fluorine, chlorine, bromine, iodine), C1-C30 alkyl, C1-C30 alkoxy;

[0017] R4 is selected from hydrogen, CF3, halogen (fluorine, chlorine, bromine, iodine), C1-C30 alkyl, C1-C30 alkoxy;

[0018] R5 is selected from hydrogen, CF3, halogen (fluorine, chlorine, bromine, iodine), C1-C30 alkyl, C1-C30 alkoxy;

[0019] R6 is selected from hydrogen, CF3, halogen (fluorine, chlorine, bromine, iodine), C1-C30 alkyl, C1-C30 alkoxy;

[0020] R7 is selected from hydrogen, CF3, halogen (fluorine, chlorine, bromine, iodine), C1-C30 alkyl, C1-C30 alkoxy;

[0021] R8 is selected from hydrogen, CF3, halogen (fluorine, chlorine, bromine, iodine), C1-C30 alkyl, C1-C30 alkoxy;

[0022] R9 is selected from hydrogen, CF3, halogen (fluorine, chlorine, bromine, iodine), C1-C30 alkyl, C1-C30 alkoxy;

[0023] R 10 Selected from hydrogen, CF3, halogen (fluorine, chlorine, bromine, iodine), C1-C30 alkyl, C1-C30 alkoxy;

[0024] R 11 Selected from hydrogen, CF3, halogen (fluorine, chlorine, bromine, iodine), C1-C30 alkyl, C1-C30 alkoxy;

[0025] R 12 Selected from hydrogen, CF3, halogen (fluorine, chlorine, bromine, iodine), C1-C30 alkyl, C1-C30 alkoxy;

[0026] R 13 Selected from hydrogen, CF3, halogen (fluorine, chlorine, bromine, iodine), C1~C30 alkyl, C1~C30 alkoxy.

[0027] More preferably, in the amidine sulfonamide derivative,

[0028] R1 is selected from hydrogen, CF3, fluorine, chlorine, bromine, methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, n-pentyl, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, tert-butoxy, n-pentyl;

[0029] R2 is selected from hydrogen, CF3, fluorine, chlorine, bromine, methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, n-pentyl, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, tert-butoxy, n-pentyloxy;

[0030] R3 is selected from hydrogen, CF3, fluorine, chlorine, bromine, methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, n-pentyl, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, tert-butoxy, n-pentyloxy;

[0031] R4 is selected from hydrogen, CF3, fluorine, chlorine, bromine, methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, n-pentyl, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, tert-butoxy, n-pentyloxy;

[0032] R5 is selected from hydrogen, CF3, fluorine, chlorine, bromine, methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, n-pentyl, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, tert-butoxy, n-pentyloxy;

[0033] R6 is selected from hydrogen, CF3, fluorine, chlorine, bromine, methyl, ethyl, methoxy, ethoxy;

[0034] R7 is selected from hydrogen, CF3, fluorine, chlorine, bromine, methyl, ethyl, methoxy, ethoxy;

[0035] R8 is selected from hydrogen, CF3, fluorine, chlorine, bromine, methyl, ethyl, methoxy, ethoxy;

[0036] R9 is selected from hydrogen, CF3, fluorine, chlorine, bromine, methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, n-pentyl, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, tert-butoxy, n-pentyloxy;

[0037] R 10 is selected from hydrogen, CF3, fluorine, chlorine, bromine, methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, n-pentyl, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, tert-butoxy, n-pentyl;

[0038] R 11 is selected from hydrogen, CF3, fluorine, chlorine, bromine, methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, n-pentyl, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, tert-butoxy, n-pentyl;

[0039] R 12 is selected from hydrogen, CF3, fluorine, chlorine, bromine, methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, n-pentyl, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, tert-butoxy, n-pentyl;

[0040] R 13 Selected from hydrogen, CF3, fluorine, chlorine, bromine, methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, n-pentyl, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, tert-butoxy, n-pentyl.

[0041] Most preferably, the structure of the amidinosulfonamide derivative is selected from one of the following structures:

[0042]

[0043]

[0044] The pharmaceutically acceptable salt refers to a salt formed by the reaction of an amidinosulfonamide derivative with a pharmaceutically acceptable inorganic acid or organic acid; wherein the inorganic acid is at least one of hydrochloric acid, hydrobromic acid, phosphoric acid, nitric acid or sulfuric acid; and the organic acid is at least one of formic acid, acetic acid, propionic acid, succinic acid, 1,5-naphthalenedisulfonic acid, asiatic acid, carbenoxolone acid, glycyrrhetinic acid, oleanolic acid, hawthorn acid, ursolic acid, corosolic acid, betulinic acid, boswellic acid, oxalic acid, tartaric acid, lactic acid, salicylic acid, benzoic acid, valeric acid, diethylacetic acid, malonic acid, succinic acid, fumaric acid, pimelic acid, adipic acid, maleic acid, malic acid, aminosulfonic acid, phenylpropionic acid, gluconic acid, ascorbic acid, nicotinic acid, isonicotinic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, citric acid or amino acids.

[0045] The second aspect of the present invention provides a pharmaceutical composition, wherein the pharmaceutical composition comprises the amidinosulfonamide derivative or a pharmaceutically acceptable salt thereof as the sole active ingredient.

[0046] The mass percentage of the amidinosulfonamide derivative or its pharmaceutically acceptable salt is 0.1-99wt%.

[0047] The third aspect of the present invention provides a pharmaceutical preparation, which is prepared from the amidinosulfonamide derivative or its pharmaceutically acceptable salt and excipients commonly used in pharmacy.

[0048] The dosage form of the pharmaceutical preparation is selected from at least one of injection, injection emulsion, tablet, pill, capsule, ointment, cream, patch, liniment, powder, spray, implant, drop, suppository, ointment or nano preparation, wherein: the injection is at least one of small volume injection, medium volume injection or large volume injection, and the nano preparation is liposome.

[0049] The fourth aspect of the present invention provides a use of the amidinosulfonamide derivative or a pharmaceutically acceptable salt thereof in the preparation of an NLRP3 inhibitor.

[0050] The fifth aspect of the present invention provides a use of the amidinosulfonamide derivative or a pharmaceutically acceptable salt thereof in the preparation of an anti-inflammatory drug.

[0051] The sixth aspect of the present invention provides a use of the amidinosulfonamide derivative or a pharmaceutically acceptable salt thereof in the preparation of a drug for treating rheumatoid arthritis or a drug for treating ulcerative bowel disease.

[0052] Due to the adoption of the above technical solution, the present invention has the following advantages and beneficial effects:

[0053] The amidine sulfonamide derivative provided by the invention not only has excellent anti-inflammatory activity but also has low cytotoxicity.

[0054] The amidinosulfonamide derivatives provided by the present invention have anti-inflammatory activity. In addition to being used in anti-inflammatory drugs, they can also be used as active ingredients in the treatment of inflammatory diseases, the treatment of nervous system diseases or the preparation of anti-tumor drugs. The amidinosulfonamide derivatives used as pharmaceutical compositions in the treatment of inflammatory diseases, the treatment of nervous system diseases or the preparation of anti-tumor drugs can be in solid form or liquid form. DETAILED DESCRIPTION

[0055] In order to explain the present invention more clearly, the present invention is further described below in conjunction with preferred embodiments. It should be understood by those skilled in the art that the following specific description is illustrative rather than restrictive, and should not be used to limit the scope of protection of the present invention.

[0056] The experimental methods in the following examples without specifying specific conditions are usually carried out under conventional conditions or under conditions recommended by the manufacturers. All raw materials without specifying the synthesis method were purchased from manufacturers such as Exploration Platform, Bidrich, and Sigma-Aldrich, and the purity was analytical grade.

[0057] Example 1

[0058] A method for preparing compound Ia comprises the following steps:

[0059]

[0060] Add 2mL acetone and ammonium thiocyanate (2mmol, 386mg) to the flask, stir and dissolve, add benzoyl chloride (2mmol, 281mg), reflux for 5 minutes, add compound 1 (2mmol, 346mg), and continue reflux for 30 minutes. After the reaction is completed, pour the reaction solution into ice water, filter, wash the filter cake with acetic acid, and dry to obtain a yellow-brown solid compound 2 with a yield of 91%.1 H NMR (600MHz, DMSO-d6) δ9.83(s,1H),7.97(d,J=7.5Hz,2H),7.58(t,J=7.4Hz,1H),7.52(t,J= 7.6Hz, 2H), 7.01 (s, 1H), 2.85 (t, J = 7.3Hz, 4H), 2.73 (t, J = 7.3Hz, 4H), 1.99 (p, J = 7.4Hz, 4H).

[0061] Compound 2 (1 mmol, 336 mg) and sodium hydroxide (4.25 mmol, 170 mg) were dissolved in 2 mL of water and refluxed for 2 h. After the reaction was completed, the mixture was cooled to 0 ° C, filtered, and the filter cake was washed with a small amount of ice water and dried to obtain a crude product. The crude product was purified by column chromatography (PE: EA = 9: 1) to obtain a white solid compound 3 with a yield of 52%. 1 H NMR (600MHz, DMSO-d6) δ9.96(s,1H),9.17(s,1H),6.99(s,1H),2.81(t,J=7.3Hz,4H),2.71(t,J=5.7Hz,4H),1.98(p,J=14.9,7.5Hz,4H).

[0062] 4 mL of ethanol, compound 3 (1 mmol, 232 mg) and iodomethane (1 mmol, 142 mg) were added to the flask and refluxed for 3 h. After the reaction, the solvent was evaporated under reduced pressure to obtain yellow solid compound 4, which was directly used in the next step. 1 H NMR (600MHz, CDCl3) δ10.22(s,1H),9.40(s,1H),7.98(s,1H),7.15(s,1H),2.98–2.80(m,8H),2.72(s,3H),2.14–2.07(m,4H).

[0063] Compound 4 (1 mmol, 246 mg) was dissolved in 2 mL DCM, triethylamine (2 mmol, 202 mg) and p-trifluoromethylbenzenesulfonyl chloride (1 mmol, 244.6 mg) were added, and the mixture was reacted at room temperature overnight. After the reaction, the mixture was purified by column chromatography (PE:EA=9:1) to obtain a white solid compound IIIa with a yield of 19.8%. 1 H NMR (600MHz, CDCl3) δ9.43 (s, 1H), 8.13 (d, J = 8.3Hz, 2H), 7.81 (d, J = 8.4Hz, 2H), 7.14(s,1H),2.91(t,J=7.3Hz,4H),2.76(s,4H),2.31(s,3H),2.13–2.03(m,4H).

[0064] Compound IIIa (0.2mmol, 91mg) was dissolved in 5mL of ethanol, triethylamine (2mmol, 202mg) and hydroxylamine hydrochloride (1mmol, 69.5mg) were added, and the mixture was refluxed for 18h. After the reaction was completed, the solvent was evaporated under reduced pressure, and the residue was dissolved in DCM (20mL) and washed with saturated brine (20mL). The organic phase was dried, filtered, and the filtrate was evaporated to dryness under reduced pressure. The mixture was purified by column chromatography (PE:EA=9:1) to obtain a white solid compound Ia with a yield of 36%. 1 H NMR(600MHz,DMSO-d6)δ9.80(s,1H),9.71(s,1H),9.08(s,1H),7.90–7.86(m,4H ),6.98(s,1H),2.80(t,J=7.4Hz,4H),2.50–2.46(m,4H),1.88(p,J=7.4Hz,4H). 13 C NMR(75MHz,DMSO-d6)δ155.99,148.23,143.37,139.59,131.85,131.43,129.28,127 .20,126.23,126.18,119.35,32.90,30.23,25.34.HRMS(ESI):m / z[M+H]+calculated for C 20 H 20 F3N3O3S:440.1255; found:440.1250.

[0065] Example 2

[0066] A method for preparing compound Ib comprises the following steps:

[0067]

[0068] Referring to the preparation method of compound Ia in Example 1, p-trifluoromethylbenzenesulfonyl chloride was replaced by benzenesulfonyl chloride to obtain white solid compound Ib with a yield of 29%. 1 H NMR(600MHz,DMSO-d6)δ9.75(s,1H),9.50(s,1H),8.98(s,1H),7.74–7.67(m,2H),7.56–7.52(m,1H),7 .49(t,J=7.5Hz,2H),6.98(s,1H),2.80(t,J=7.4Hz,4H),2.53(t,J=7.3Hz,4H),1.90(p,J=7.4Hz,4H). 13C NMR(151MHz,DMSO-d6)δ156.10,144.28,143.35,139.59,131.80,129.39,128.95,126.28,119.24,32.92,30.28,25.40.HRMS(ESI):m / z[M+H]+calculated for C 19 H 21 N3O3S:372.1382; found:372.1374.

[0069] Example 3

[0070] A method for preparing compound Ic comprises the following steps:

[0071]

[0072] Referring to the preparation method of compound Ia in Example 1, p-fluorobenzenesulfonyl chloride was used instead of p-trifluoromethylbenzenesulfonyl chloride to obtain white solid compound Ic with a yield of 40%. 1 H NMR(600MHz,DMSO-d6)δ9.76(s,1H),9.56(s,1H),9.00(s,1H),7.76(dd,J=8.7,5.4Hz,2H),7.33( t,J=8.8Hz,2H),6.98(s,1H),2.80(t,J=7.3Hz,4H),2.52(t,J=7.3Hz,4H),1.90(p,J=7.4Hz,4H). 13 C NMR(151MHz,DMSO-d6)δ164.81,163.16,156.00,143.37,140.80,139.57,129.36,129.19 ,129.13,119.27,116.02,115.88,32.92,30.29,25.39.HRMS(ESI):m / z[M+H]+calculated for C 19 H 20 FN3O3S:390.1287; found:390.1282.

[0073] Example 4

[0074] A method for preparing compound Id comprises the following steps:

[0075]

[0076] Referring to the preparation method of compound Ia in Example 1, p-trifluoromethylbenzenesulfonyl chloride was replaced with m-fluorobenzenesulfonyl chloride to obtain white solid compound Id. 1H NMR(600MHz,DMSO-d6)δ9.79(s,1H),9.64(s,1H),9.04(s,1H),7.58–7.52(m,2H),7.49–7.45(m,1H),7.42–7.38 (m,1H),6.99(s,1H),2.81(t,J=7.4Hz,4H),2.53(t,J=7.4Hz,4H),1.91(p,J=7.5Hz,4H).HRMS(ESI):m / z[M+Na] + calculated for C 19 H 20 FN3O3S:412.1107; found:412.1102.

[0077] Example 5

[0078] A method for preparing compound Ie comprises the following steps:

[0079]

[0080] Referring to the preparation method of compound Ia in Example 1, p-trifluoromethylbenzenesulfonyl chloride was replaced with o-fluorobenzenesulfonyl chloride to obtain white solid compound Ie with a yield of 32%. 1 H NMR (600MHz, DMSO-d6) δ9.82(s,1H),9.54(s,1H),9.06(s,1H),7.77(dd,J=7.5,5.9Hz,1H),7.59(dd,J=12.3,6.4Hz,1H),7.34–7.30(m,1 H),7.30–7.27(m,1H),6.99(s,1H),2.81(t,J=7.4Hz,4H),2.60(t,J=7.5Hz,4H),1.92(p,J=7.5Hz,4H).HRMS(ESI):m / z[M+H]+calculated for C 19 H 20 FN3O3S:390.1288; found:390.1282.

[0081] Example 6

[0082] A method for preparing a compound If comprises the following steps:

[0083]

[0084] Referring to the preparation method of compound Ia in Example 1, p-chlorobenzenesulfonyl chloride was used instead of p-trifluoromethylbenzenesulfonyl chloride to obtain white solid compound If with a yield of 42%. 1H NMR(600MHz,DMSO-d6)δ9.77(s,1H),9.58(s,1H),9.02(s,1H),7.71–7.69(m,2H),7.58–7 .55(m,2H),6.98(s,1H),2.80(t,J=7.3Hz,4H),2.54–2.51(m,4H),1.90(p,J=7.5Hz,4H). 13 C NMR(151MHz,DMSO-d6)δ155.99,143.37,139.58,136.51,129.33,129.05,128.29,119.29,32.92,30.29,25.38.HRMS(ESI):m / z[M+H] + calculated for C 19 H 20 ClN3O3S:406.0992; found:406.0987.

[0085] Example 7

[0086] A method for preparing compound Ig, comprising the following steps:

[0087]

[0088] Referring to the preparation method of compound Ia in Example 1, p-methoxybenzenesulfonyl chloride was used instead of p-trifluoromethylbenzenesulfonyl chloride to obtain white solid compound Ig with a yield of 35%. 1 H NMR (600MHz, DMSO-d6) δ9.75(s,1H),9.39(s,1H),8.90(s,1H),7.64(t,J=5.9Hz,2H),7.02–6.99(m,2H),6.97( s,1H),3.81(s,3H),2.80(t,J=7.4Hz,4H),2.54(t,J=7.4Hz,4H),1.90(p,J=7.5Hz,4H).HRMS(ESI):m / z[M+Na] + calculated for C 20 H 23 N3O4S:424.1307; found:424.1301.

[0089] Example 8

[0090] A method for preparing compound Ih comprises the following steps:

[0091]

[0092] Referring to the preparation method of compound Ia in Example 1, thiophenesulfonyl chloride was used instead of trifluoromethylbenzenesulfonyl chloride to obtain white solid compound Ih in a yield of 25%. 1 H NMR(600MHz,DMSO-d6)δ9.47(s,1H),8.01(d,J=5.0Hz,1H),7.67(d,J=3.7Hz,1H),7.19 (t,J=4.4Hz,1H),7.08(s,1H),2.85–2.75(m,6H),2.42–2.34(m,2H),1.91–1.79(m,4H). 13 C NMR(151MHz,DMSO-d6)δ148.41,144.52,143.15,141.17,134.68,134.57,130.00,127.79,121.60,32.84,31.04,25.66.HRMS(ESI):m / z[M+H]+calculated for C 17 H 19 N3O3S2:378.0946; found:378.0941.

[0093] Example 9

[0094] A method for preparing compound Ii comprises the following steps:

[0095]

[0096] Referring to the preparation method of compound Ia in Example 1, (E)-phenylvinylsulfonyl chloride was used instead of trifluoromethylbenzenesulfonyl chloride to obtain white solid compound Ii with a yield of 37%. 1 H NMR (600MHz, DMSO-d6) δ8.98(s,1H),7.48(d,J=2.3Hz,1H),7.32(dt,J=14.9,7.3Hz,4H),7.26(t,J=7.0Hz,1H),6.99(s,1H) ,5.94(s,1H),2.85(dd,J=16.8,9.5Hz,4H),2.80(t,J=7.4Hz,4H),1.94(p,J=7.5Hz,4H).HRMS(ESI):m / z[M+H]+calculated for C 21 H 23 N3O3S:398.1539; found:398.1533.

[0097] Example 10

[0098] A method for preparing a pharmaceutically acceptable salt of the compound prepared in Examples 1 to 9:

[0099] The compounds prepared in Examples 1 to 9 are reacted with pharmaceutically acceptable inorganic acids or organic acids to obtain pharmaceutically acceptable salts of the corresponding amidinosulfonamide derivatives.

[0100] In the preparation process of the pharmaceutical salt of the amidine sulfonamide derivative, the inorganic acid is at least one of hydrochloric acid, hydrobromic acid, phosphoric acid, nitric acid or sulfuric acid; the organic acid is at least one of formic acid, acetic acid, propionic acid, succinic acid, 1,5-naphthalene disulfonic acid, asiatic acid, carbenoxolone acid, glycyrrhetinic acid, oleanolic acid, hawthorn acid, ursolic acid, corosolic acid, betulinic acid, boswellic acid, oxalic acid, tartaric acid, lactic acid, salicylic acid, benzoic acid, valeric acid, diethylacetic acid, malonic acid, succinic acid, fumaric acid, pimelic acid, adipic acid, maleic acid, malic acid, aminosulfonic acid, phenylpropionic acid, gluconic acid, ascorbic acid, nicotinic acid, isonicotinic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, citric acid or amino acid.

[0101] Embodiment 11

[0102] A pharmaceutical composition prepared from amidinosulfonamide derivatives:

[0103] A therapeutically effective amount of the compounds prepared in Examples 1 to 9 of the present invention is mixed or dissolved with one or more pharmaceutically acceptable carriers, excipients and auxiliary materials to obtain a corresponding pharmaceutical composition.

[0104] A pharmaceutical preparation prepared from amidinosulfonamide derivatives:

[0105] The compounds prepared in Examples 1 to 9 of the present invention are respectively subjected to corresponding operations according to the preparation methods of existing pharmaceutical preparations to prepare self-injections, injectable emulsions, tablets, pills, capsules, ointments, creams, patches, liniments, powders, sprays, implants, drops, suppositories, ointments or nano preparations.

[0106] Example 12

[0107] Cytotoxicity test and in vitro anti-inflammatory activity test of amidinosulfonamide derivatives:

[0108] The conventional CCK-8 method was used to test the cytotoxicity of the compounds prepared in Examples 1 to 9, wherein: the test cell line was BMDM macrophages extracted from the bone marrow of C57BL / 6J mice from Hangzhou Ziyuan Experimental Animal Technology Co., Ltd. The cells extracted from the bone marrow of mice were treated with M-CSF cell stimulating factor for 7 days, the cells were replaced with medium on the 3rd and 5th days, and matured into macrophages on the 7th day. The culture medium was RPMI1640 medium containing 10% fetal bovine serum, 1% penicillin-streptomycin mixture and 0.05 mM β-mercaptoethanol.

[0109] Sample preparation: The target compound was prepared into a stock solution using cell culture grade DMSO solvent. When used, the compound was prepared into a stock solution with the corresponding culture medium at a concentration of 0.1uM, 1uM and 100uM. The solution was prepared and used immediately.

[0110] The experimental method is: bone marrow BMDM cells are extracted from the femur and tibia of male C57BL / 6J mice of about 6 weeks old, and the cells are cultured in PRMI 1640 complete medium containing 20ng / mL for 7 days to mature, and the BMDM cells are seeded in 96-well plates and incubated overnight. Then the cells are treated with different concentrations (starting concentration is 100uM, 2-fold or 3-fold dilution) of the target compound for 24h. Under light-proof conditions, the culture supernatant is replaced with a mixed solution containing CCK-8 and basal culture medium, and incubated in a cell culture incubator for about 1h. The OD value is measured using an enzyme reader, and the drug concentration (CC50) that causes 50% cell death is simulated and calculated by GraphPad Prism software.

[0111] The ELISA method was used to detect the secretion level of human IL-1β. The compounds prepared in Examples 1 to 9 were tested for in vitro anti-inflammatory activity, wherein: Mouse bone marrow cell extraction: 6-8 week old male C57BL / 6J mice were killed by cervical dislocation, and disinfected by soaking in 75% alcohol for 5 minutes, and the femur and tibia of the mice were removed, the skin and muscle tissue were removed, and the cells were moved into cold PBS for soaking. Cut off the joint connection at both ends, and the bone marrow cavity was rinsed 3 times with sterile phosphate buffer PBS. The cells were filtered with a 70μM cell filter and collected into a 50mL centrifuge tube, and centrifuged at 1600r / min for 3 minutes; the supernatant was discarded, and the red blood cell lysis solution was added for 2 minutes, and the cells were centrifuged at 1600r / min for 3 minutes, and the supernatant was discarded, and the cells were resuspended.

[0112] Mouse bone marrow cell culture: Use M-CSF containing 20 ng / mL for culture, change the medium on the third and fifth days, and allow the cells to mature on the seventh day.

[0113] NLRP3 inflammasome activation: After the mouse bone marrow cells matured, they were digested with trypsin for about 5 minutes, collected, centrifuged to remove the supernatant, resuspended, and concentrated to 1×10 5 Cells were seeded in 96-well plates at a concentration of 100 μL cell / mL overnight, and stimulated with 50 ng / mL LPS for 3 h. The culture medium was removed, and DMSO prepared in RPMI 1640 basal culture medium and 100 μL of the sample to be tested (concentrations of 0.1 μM or 1 μM, respectively) were added for 30 min. Without removing the culture medium, ATP with a final concentration of 5 mM was added for 1 h.

[0114] ELISA method for detecting IL-1β level: The supernatant was taken for ELISA detection, and the content of each index was calculated according to the absorbance. The test results are shown in Table 1, wherein the test samples are the compounds prepared in Examples 1-9.

[0115] Table 1

[0116]

[0117]

[0118] From the data listed in Table 1, we can see that:

[0119] The compounds prepared in Examples 1 to 9 of the present invention have good anti-inflammatory activity and show a certain inhibitory effect on IL-1β at two concentrations. More importantly, the compounds prepared in Examples 1 to 9 of the present invention have low cytotoxicity. Except for compounds Ia, Id, and Ig, CC 50 The values ​​were all greater than or equal to 100 μM, and the cytotoxicity of compounds Ia, Id, and Ig were also lower than that of Rubescensine A.

[0120] When the compounds prepared in Examples 1 to 9 of the present invention were at a low concentration of 0.1 μM, the inhibition rates of compounds Ib, Ic and Id on IL-1β were more than 50%. When the concentration was high at 1 μM, except for Ic and Ii, the inhibition rates were all more than 50%, among which compound Id had the highest inhibitory activity, reaching 69.5±21.4%.

[0121] In summary, we can see that:

[0122] First, the amidinosulfonamide derivatives provided by the present invention have higher anti-inflammatory activity and lower cytotoxicity than oridonin, and can therefore be used as active ingredients in the preparation of inflammatory diseases, nervous system diseases or anti-tumor drugs; secondly, the amidinosulfonamide derivatives provided by the present invention provide new ideas and methods for preparing new drugs with better efficacy and lower toxic side effects, and have broad application prospects.

[0123] The above is only a preferred embodiment of the present invention, and does not limit the present invention in any form. Although the present invention has been disclosed as a preferred embodiment, it is not used to limit the present invention. Any technician familiar with this patent can make some changes or modify the technical contents suggested above into equivalent embodiments without departing from the scope of the technical solution of the present invention. However, any simple modification, equivalent change and modification made to the above embodiments according to the technical essence of the present invention without departing from the content of the technical solution of the present invention still fall within the scope of the solution of the present invention.

Claims

1. An amidinosulfonamide derivative or a pharmaceutically acceptable salt thereof, characterized in that: The general structural formula is shown below: in: R is selected from one of the following structures: R1 is selected from hydrogen, CF3, halogen, C1-C30 alkyl, C1-C30 alkoxy; R2 is selected from hydrogen, CF3, halogen, C1-C30 alkyl, C1-C30 alkoxy; R3 is selected from hydrogen, CF3, halogen, C1-C30 alkyl, C1-C30 alkoxy; R4 is selected from hydrogen, CF3, halogen, C1-C30 alkyl, C1-C30 alkoxy; R5 is selected from hydrogen, CF3, halogen, C1-C30 alkyl, C1-C30 alkoxy; R6 is selected from hydrogen, CF3, halogen, C1-C30 alkyl, C1-C30 alkoxy; R7 is selected from hydrogen, CF3, halogen, C1-C30 alkyl, C1-C30 alkoxy; R8 is selected from hydrogen, CF3, halogen, C1-C30 alkyl, C1-C30 alkoxy; R9 is selected from hydrogen, CF3, halogen, C1-C30 alkyl, C1-C30 alkoxy; R 10 Selected from hydrogen, CF3, halogen, C1-C30 alkyl, C1-C30 alkoxy; R 11 Selected from hydrogen, CF3, halogen, C1-C30 alkyl, C1-C30 alkoxy; R 12 Selected from hydrogen, CF3, halogen, C1-C30 alkyl, C1-C30 alkoxy; R 13 Selected from hydrogen, CF3, halogen, C1~C30 alkyl, C1~C30 alkoxy.

2. The amidinosulfonamide derivative or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: In the amidinosulfonamide derivative, R1 is selected from hydrogen, CF3, fluorine, chlorine, bromine, methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, n-pentyl, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, tert-butoxy, n-pentyl; R2 is selected from hydrogen, CF3, fluorine, chlorine, bromine, methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, n-pentyl, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, tert-butoxy, n-pentyloxy; R3 is selected from hydrogen, CF3, fluorine, chlorine, bromine, methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, n-pentyl, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, tert-butoxy, n-pentyloxy; R4 is selected from hydrogen, CF3, fluorine, chlorine, bromine, methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, n-pentyl, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, tert-butoxy, n-pentyloxy; R5 is selected from hydrogen, CF3, fluorine, chlorine, bromine, methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, n-pentyl, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, tert-butoxy, n-pentyloxy; R6 is selected from hydrogen, CF3, fluorine, chlorine, bromine, methyl, ethyl, methoxy, ethoxy; R7 is selected from hydrogen, CF3, fluorine, chlorine, bromine, methyl, ethyl, methoxy, ethoxy; R8 is selected from hydrogen, CF3, fluorine, chlorine, bromine, methyl, ethyl, methoxy, ethoxy; R9 is selected from hydrogen, CF3, fluorine, chlorine, bromine, methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, n-pentyl, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, tert-butoxy, n-pentyloxy; R 10 is selected from hydrogen, CF3, fluorine, chlorine, bromine, methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, n-pentyl, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, tert-butoxy, n-pentyl; R 11 is selected from hydrogen, CF3, fluorine, chlorine, bromine, methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, n-pentyl, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, tert-butoxy, n-pentyl; R 12 is selected from hydrogen, CF3, fluorine, chlorine, bromine, methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, n-pentyl, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, tert-butoxy, n-pentyl; R 13 Selected from hydrogen, CF3, fluorine, chlorine, bromine, methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, n-pentyl, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, tert-butoxy, n-pentyl.

3. The amidinosulfonamide derivative or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: The structure of the amidinosulfonamide derivative is selected from one of the following structures:

4. A pharmaceutical composition, characterized in that The pharmaceutical composition comprises the amidinosulfonamide derivative or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 3 as the sole active ingredient.

5. A pharmaceutical preparation, characterized in that The pharmaceutical preparation is prepared from the amidinosulfonamide derivative or its pharmaceutically acceptable salt according to any one of claims 1 to 3 and excipients commonly used in pharmacy.

6. The pharmaceutical preparation according to claim 5, characterized in that The dosage form of the pharmaceutical preparation is selected from at least one of injection, injection emulsion, tablet, pill, capsule, ointment, cream, patch, liniment, powder, spray, implant, drop, suppository, ointment or nano preparation.

7. Use of the amidinosulfonamide derivative or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 3 in the preparation of an NLRP3 inhibitor.

8. Use of the amidinosulfonamide derivative or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 3 in the preparation of anti-inflammatory drugs.

9. Use of the amidinosulfonamide derivative or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 3 in the preparation of a drug for treating rheumatoid arthritis or a drug for treating ulcerative bowel disease.

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