Amidinyl sulfonamide derivatives and uses thereof

Abstract

The application discloses an amidine sulfonamide derivative or a pharmaceutically acceptable salt thereof, and a structure general formula is as follows: The amidine sulfonamide derivative provided by the application has excellent anti-inflammatory activity and low toxicity.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, specifically relating to an amidosulfonamide derivative and its applications. Background Technology

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

[0003]

[0004] In mouse models, MCC950 effectively reduced the severity of experimental autoimmune encephalomyelitis, thus demonstrating its potential application 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 serum liver enzyme levels, leading to a lack of further progress in clinical development.

[0005] Inflammation driven by aberrant activation of NLRP3 is the basis of many chronic degenerative diseases, including type II diabetes, gout, atherosclerosis, inflammatory bowel disease, and Alzheimer's disease. Therefore, NLRP3 inhibition has become a potential therapeutic strategy for many diseases. Among these, 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 significant progress, with small molecule inhibitors of various structural types reported. Some highly active small molecule compounds, such as IFM-2427, Somalix, dapansutrile, and NT-0167, have entered clinical trials, but none have yet been approved for marketing. Therefore, it is necessary to continue developing NLRP3 inhibitors with novel structures. Summary of the Invention

[0006] The purpose of this invention is to provide an amidosulfonamide derivative.

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

[0008] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0009] In a first aspect, the present invention provides an amidine sulfonamide derivative or a pharmaceutically acceptable salt thereof, having the following general structural formula:

[0010]

[0011] in:

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

[0013]

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

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

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

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

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

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

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

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

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

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

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

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

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

[0027] Preferably, in the amidosulfonamide 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, and n-pentoxy.

[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, and n-pentoxy.

[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, and n-pentoxy.

[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, and n-pentoxy.

[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, and n-pentoxy.

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

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

[0035] R8 is selected from hydrogen, CF3, fluorine, chlorine, bromine, methyl, ethyl, methoxy, and 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, and n-pentoxy.

[0037] R 10 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, and n-pentoxy;

[0038] R 11 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, and n-pentoxy;

[0039] R 12 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, and n-pentoxy;

[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, and n-pentoxy.

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

[0042]

[0043]

[0044] The pharmaceutical salt refers to the salt formed by the reaction of amidine sulfonamide derivatives with a pharmaceutically acceptable inorganic 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, linalool, glycyrrhizic 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, sulfamic acid, phenylpropionic acid, gluconic acid, ascorbic acid, nicotinic acid, isonicotinic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, citric acid, or amino acids.

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

[0046] The mass percentage of the amidosulfonamide derivative or its pharmaceutical salt is 0.1-99 wt%.

[0047] In a third aspect, the present invention provides a pharmaceutical preparation made from the amidosulfonamide derivative or a pharmaceutically acceptable salt thereof and pharmaceutically commonly used excipients.

[0048] The dosage form of the pharmaceutical preparation is selected from at least one of the following: injection, emulsion for injection, tablet, pill, capsule, ointment, cream, patch, liniment, powder, spray, implant, drops, 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] In a fourth aspect, the present invention provides the use of the said amidosulfonamide derivative or a pharmaceutical salt thereof in the preparation of an NLRP3 inhibitor.

[0050] A fifth aspect of the invention provides the use of the said amidosulfonamide derivative or a pharmaceutical salt thereof in the preparation of an anti-inflammatory medicament.

[0051] In a sixth aspect, the present invention provides the use of the aforementioned amidosulfonamide derivative or a pharmaceutical salt thereof in the preparation of a medicament for treating rheumatoid arthritis and a medicament for treating ulcerative colitis.

[0052] By adopting the above technical solution, the present invention has the following advantages and beneficial effects:

[0053] The amidosulfonamide derivatives provided by this invention not only have excellent anti-inflammatory activity, but also low cytotoxicity.

[0054] The amidosulfonamide derivative provided by this invention has anti-inflammatory activity. In addition to being used in anti-inflammatory drugs, it can also be used as an active ingredient in the preparation of drugs for the treatment of inflammatory diseases, the treatment of nervous system diseases, or anti-tumor drugs. Furthermore, when used as a pharmaceutical composition in the preparation of drugs for the treatment of inflammatory diseases, the treatment of nervous system diseases, or anti-tumor drugs, it can be in solid or liquid form. Detailed Implementation

[0055] To more clearly illustrate the present invention, the following description, in conjunction with preferred embodiments, further clarifies the invention. Those skilled in the art should understand that the specific descriptions below are illustrative rather than restrictive, and should not be construed as limiting the scope of protection of the present invention.

[0056] Unless otherwise specified, experimental methods in the following examples were performed under standard conditions or as recommended by the manufacturer. All raw materials without specified synthesis methods were purchased from manufacturers such as Exploration Platform, Bitmain, and Sigma-Aldrich, and were of analytical grade.

[0057] Example 1

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

[0059]

[0060] Add 2 mL of acetone and ammonium thiocyanate (2 mmol, 386 mg) to a flask, stir to dissolve, then add benzoyl chloride (2 mmol, 281 mg), reflux for 5 minutes, then add compound 1 (2 mmol, 346 mg), and continue reflux for 30 minutes. After the reaction is complete, pour the reaction solution into ice water, filter, wash the filter cake with acetic acid, and dry to give a yellowish-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 complete, 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 the crude product. The crude product was purified by column chromatography (PE:EA = 9:1) to give compound 3 as a white solid, 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] Add 4 mL of ethanol, compound 3 (1 mmol, 232 mg), and iodomethane (1 mmol, 142 mg) to a flask, and reflux for 3 h. After the reaction is complete, evaporate the solvent under reduced pressure to obtain a yellow solid compound 4, which can be used directly in the next reaction. 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 of LCM, and triethylamine (2 mmol, 202 mg) and p-trifluoromethylbenzenesulfonyl chloride (1 mmol, 244.6 mg) were added. The mixture was reacted overnight at room temperature. After the reaction was complete, the solution was purified by column chromatography (PE:EA = 9:1) to give a white solid compound IIIa in 19.8% yield. 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 (0.2 mmol, 91 mg) was dissolved in 5 mL of ethanol, and triethylamine (2 mmol, 202 mg) and hydroxylamine hydrochloride (1 mmol, 69.5 mg) were added. The mixture was refluxed for 18 h. After the reaction was complete, the solvent was evaporated under reduced pressure, and the residue was dissolved in DCM (20 mL) and washed with saturated brine (20 mL). The organic phase was dried, filtered, and the filtrate was evaporated to dryness under reduced pressure. The filtrate was purified by column chromatography (PE:EA = 9:1) to give a white solid compound Ia in 36% yield. 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 includes the following steps:

[0067]

[0068] Following the preparation method of compound Ia in Example 1, p-trifluoromethylbenzenesulfonyl chloride was replaced with 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 includes the following steps:

[0071]

[0072] Following the preparation method of compound Ia in Example 1, p-fluorobenzenesulfonyl chloride was substituted for p-trifluoromethylbenzenesulfonyl chloride to obtain white solid compound Ic in 40% yield. 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 includes the following steps:

[0075]

[0076] Following the preparation method of compound Ia in Example 1, p-trifluoromethylbenzenesulfonyl chloride was replaced with m-fluorobenzenesulfonyl chloride to obtain a 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 includes the following steps:

[0079]

[0080] Following the preparation method of compound Ia in Example 1, o-fluorobenzenesulfonyl chloride was substituted for p-trifluoromethylbenzenesulfonyl chloride to obtain white solid compound Ie in 32% yield. 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 includes the following steps:

[0083]

[0084] Following the preparation method of compound Ia in Example 1, p-chlorobenzenesulfonyl chloride was substituted for p-trifluoromethylbenzenesulfonyl chloride to obtain a 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 includes the following steps:

[0087]

[0088] Following the preparation method of compound Ia in Example 1, p-methoxybenzenesulfonyl chloride was substituted for p-trifluoromethylbenzenesulfonyl chloride to obtain a 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 includes the following steps:

[0091]

[0092] Following the preparation method of compound Ia in Example 1, thiophene sulfonyl chloride was substituted for p-trifluoromethylbenzenesulfonyl chloride to obtain white solid compound Ih in 25% yield. 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 includes the following steps:

[0095]

[0096] Following the preparation method of compound Ia in Example 1, (E)-styrenesulfonyl chloride was substituted for p-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 pharmaceutical salts of the compounds prepared in Examples 1-9:

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

[0100] In the preparation of the above-mentioned pharmaceutical salts of amidosulfonamide derivatives: 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-naphthalenedisulfonic acid, linaloic acid, glycyrrhetinic 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.

[0101] Example 11

[0102] A pharmaceutical composition prepared from an amidosulfonamide derivative:

[0103] A therapeutically effective amount of the compounds prepared in Examples 1 to 9 of this invention is mixed or miscible with one or more pharmaceutically acceptable carriers, excipients, and excipients to obtain the corresponding pharmaceutical compositions.

[0104] A pharmaceutical preparation made from an amidine sulfonamide derivative:

[0105] The compounds prepared in Examples 1 to 9 of this invention are used to prepare injectable preparations, emulsions for injection, tablets, pills, capsules, ointments, creams, patches, liniments, powders, sprays, implants, drops, suppositories, ointments, or nano-preparations by performing corresponding operations according to existing pharmaceutical preparation methods.

[0106] Example 12

[0107] Cytotoxicity and in vitro anti-inflammatory activity assays of amidine sulfonamide derivatives:

[0108] The compounds prepared in Examples 1-9 were subjected to cytotoxicity tests using the conventional CCK-8 assay. The experimental cell line used was BMDM macrophages extracted from the bone marrow of C57BL / 6J mice from Hangzhou Ziyuan Experimental Animal Technology Co., Ltd. The cells were treated with M-CSF cell stimulating factor for 7 days, with the medium changed on days 3 and 5, and maturing into macrophages on day 7. The culture medium was RPMI 1640 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 using the sample, the compound was prepared into concentrations of 0.1 μM, 1 μM, and 100 μM using the corresponding culture medium. The sample was prepared and used immediately.

[0110] The experimental method was as follows: Bone marrow BMDM cells were extracted from the femur and tibia of male C57BL / 6J mice at approximately 6 weeks of age. After 7 days of maturation in PRMI 1640 complete medium containing 20 ng / mL, the BMDM cells were seeded in 96-well plates and incubated overnight. The cells were then treated with different concentrations (starting at 100 μM, 2- or 3-fold dilution) of the target compound for 24 h. Under light-protected conditions, the culture supernatant was replaced with a mixed solution containing CCK-8 and basal medium, and the cells were incubated for approximately 1 h in a cell culture incubator. OD values ​​were measured using a microplate reader, and the drug concentration (CC50) that induced 50% cell death was calculated using GraphPad Prism software.

[0111] The ELISA method was used to detect the secretion level of human IL-1β. The compounds prepared in Examples 1-9 were then subjected to in vitro anti-inflammatory activity assays. Specifically: Mouse bone marrow cell extraction: 6-8 week old male C57BL / 6J mice were euthanized by cervical dislocation. The cells were disinfected by soaking in 75% alcohol for 5 minutes. The femur and tibia were removed, and the skin and muscle tissue were removed. The cells were then soaked in cold PBS. The joint connections at both ends were cut off. The bone marrow cavity was rinsed three times with sterile phosphate-buffered saline (PBS). Cells were collected using a 70 μM cell filter and centrifuged in 50 mL centrifuge tubes at 1600 rpm for 3 minutes. The supernatant was discarded, and the cells were lysed in erythrocyte lysis buffer for 2 minutes, centrifuged at 1600 rpm for 3 minutes, and the supernatant was discarded. The cells were resuspended.

[0112] Mouse bone marrow cell culture: cultured in M-CSF containing 20 ng / mL, with medium changes on the third and fifth days, and cell maturation on the seventh day.

[0113] NLRP3 inflammasome activation: After mouse bone marrow cells matured, they were collected by trypsin digestion for about 5 minutes, centrifuged to remove the supernatant, resuspended, and then... (The sentence is incomplete and requires more context to translate accurately). 5 Cells were seeded in 96-well plates at a concentration of [cell / mL] and incubated overnight. Cells were then stimulated with 50 ng / mL LPS for 3 h. The culture medium was removed, and DMSO prepared from RPMI 1640 basal medium and 100 μL of the test sample (concentrations of 0.1 μM or 1 μM, respectively) were added for stimulation for 30 min without removing the culture medium. Finally, ATP was added to a final concentration of 5 mM for stimulation for 1 h.

[0114] ELISA detection of IL-1β levels: The supernatant was used for ELISA detection, and the content of each indicator was calculated based on the absorbance. The experimental results are shown in Table 1, where the test samples were compounds prepared in Examples 1-9.

[0115] Table 1

[0116]

[0117]

[0118] The data listed in Table 1 shows that:

[0119] The compounds prepared in Examples 1-9 of this invention exhibit good anti-inflammatory activity, showing some inhibitory effect on IL-1β at both concentrations. More importantly, the compounds prepared in Examples 1-9 of this invention have low cytotoxicity, except for compounds Ia, Id, and Ig, CC... 50 The values ​​were all greater than or equivalent to 100 μM, and the cytotoxicity of compounds Ia, Id, and Ig was also lower than that of oridonin.

[0120] The compounds prepared in Examples 1-9 of this invention exhibited inhibition rates of over 50% against IL-1β at a low concentration of 0.1 μM, with compounds Ib, Ic, and Id showing the highest inhibition rate. At a high concentration of 1 μM, all compounds except Ic and Ii achieved inhibition rates of over 50%, with compound Id showing the highest inhibitory activity at 69.5 ± 21.4%.

[0121] In conclusion, it can be seen that:

[0122] First, the amidosulfonamide derivatives provided by this invention have higher anti-inflammatory activity and lower cytotoxicity compared to oridonin, and therefore can be used as active ingredients in the preparation of drugs for inflammatory diseases, nervous system diseases, or anti-tumor diseases. Second, the amidosulfonamide derivatives provided by this invention offer new ideas and methods for preparing novel drugs with better efficacy and lower toxicity, and have broad application prospects.

[0123] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention in any way. Although the present invention has been disclosed above with reference to preferred embodiments, it is not intended to limit the present invention. Any person skilled in the art can make some modifications or alterations to the above-described technical content to create equivalent embodiments without departing from the scope of the present invention. Any simple modifications, equivalent changes, and alterations made to the above embodiments based on the technical essence of the present invention without departing from the scope of the present invention shall still fall within the scope of the present invention.

Claims

1. An amidine sulfonamide derivative or its pharmaceutical salt, characterized in that, The general structural formula is as follows: in: R is selected from one of the following structures: 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, and n-pentoxy. 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, and n-pentoxy. 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, and n-pentoxy. 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, and n-pentoxy. 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, and n-pentoxy. R6 is selected from hydrogen, CF3, fluorine, chlorine, bromine, methyl, ethyl, methoxy, and ethoxy. R7 is selected from hydrogen, CF3, fluorine, chlorine, bromine, methyl, ethyl, methoxy, and ethoxy. R8 is selected from hydrogen, CF3, fluorine, chlorine, bromine, methyl, ethyl, methoxy, and 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, and n-pentoxy. R 10 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, and n-pentoxy; R 11 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, and n-pentoxy; R 12 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, and n-pentoxy; 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, and n-pentoxy.

2. The amidine sulfonamide derivative or its pharmaceutical salt according to claim 1, characterized in that, The structure of the amidosulfonamide derivative is selected from one of the following structures:

3. A pharmaceutical composition, characterized in that, The pharmaceutical composition comprises, as the sole active ingredient, the amidosulfonamide derivative or its pharmaceutical salt as described in claim 1 or 2.

4. A pharmaceutical preparation, characterized in that, The pharmaceutical preparation is made from the amidosulfonamide derivative as described in claim 1 or 2, or its pharmaceutical salt, and pharmaceutically commonly used excipients.

5. The pharmaceutical preparation according to claim 4, characterized in that, The dosage form of the pharmaceutical preparation is selected from at least one of the following: injection, emulsion for injection, tablet, pill, capsule, ointment, cream, patch, liniment, powder, spray, implant, drops, suppository, ointment, or nano-preparation.

6. The use of an amidosulfonamide derivative of claim 1 or 2 or a pharmaceutically acceptable salt thereof in the preparation of an NLRP3 inhibitor.

7. The use of an amidosulfonamide derivative or a pharmaceutical salt thereof as described in claim 1 or 2 in the preparation of an anti-inflammatory medicament.

8. The use of an amidosulfonamide derivative or a pharmaceutical salt thereof as described in claim 1 or 2 in the preparation of a medicament for treating rheumatoid arthritis or a medicament for treating ulcerative colitis.

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