Matrine-based compounds, preparation method and application thereof

Abstract

This invention discloses matrine compounds, their preparation methods and applications, relating to the field of pharmaceutical technology. The general chemical structures of the new matrine compounds and their salts are shown in formulas (1) and (2). This invention also provides a method for preparing and synthesizing matrine derivatives and their salts. The matrine prepared by this invention has anti-inflammatory and antioxidant effects, exhibits very good activity, and has achieved good results in anti-inflammatory activity tests.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, specifically to matrine compounds, their preparation methods, and applications. Background Technology

[0002] Quinolizidine alkaloids are widely distributed in higher plants, such as those belonging to the Fabaceae, Solanaceae, Rosaceae, and Berberidaceae families. This class of alkaloids is numerous, with matrines being a representative example.

[0003] Matrine is mainly derived from medicinal plants such as Sophora flavescens, Sophora alopecuroides, and Sophora tonkinensis. Many years ago, matrine-like alkaloids were extracted and isolated from Sophora flavescens, with matrine and oxymatrine being found in high concentrations, thus attracting widespread scholarly attention. In recent years, rapid advancements in science and technology have propelled progress in separation and analysis techniques, leading researchers to discover more than 20 additional matrine-like alkaloids, such as sophoridine, oxysophoridine, sophoridine, sophoridine alcohol, and lymanin.

[0004] Matrine alkaloids have been developed into drugs for clinical use, such as matrine injection, matrine capsules, matrine tablets, and oxymatrine injection. They possess analgesic, anti-inflammatory, antitumor, cardiotonic, antiarrhythmic, anti-liver fibrosis, bactericidal, and antiviral effects.

[0005] The therapeutic effect of matrine on lung injury was studied. Lung injury was assessed by histological examination, lung dry / wet ratio, and protein content in bronchoalveolar lavage fluid. Cytokine and reactive oxygen species (ROS) production were detected by ELISA and flow cytometry. Results showed that matrine reduced the production of inflammatory mediators such as TNF-α and IL-6, improved lipopolysaccharide (LPS)-induced lung histopathological changes, and alleviated pulmonary vascular leakage and pulmonary edema. High-dose matrine significantly reduced LPS-induced mortality in mice. This suggests that matrine has anti-inflammatory and antioxidant effects. Based on the discovery by Wu Qiuye's research group (CN103936736) that M-19 (13-methylaminothiomatrine) significantly enhances the bioactivity of matrine, this invention designed and synthesized M-19 analogues and obtained a series of derivatives, achieving good results in anti-inflammatory activity tests. Summary of the Invention

[0006] The purpose of this invention is to solve the above-mentioned problems and to provide matrine compounds, their preparation methods and applications.

[0007] To achieve the above objectives, the technical solution of the present invention is as follows: Matrine compounds and their pharmaceutically acceptable salts, the general structural formula of which is shown in formula (1) or formula (2) below.

[0008]

[0009] Where X is a sulfur atom;

[0010] R1 is selected from monosubstituted, disubstituted, or polysubstituted alkyl-substituted phenyl, alkoxy-substituted phenyl, halogen-substituted phenyl, nitro-substituted phenyl, cyano-substituted phenyl, trifluoromethyl-substituted phenyl, hydroxy-substituted phenyl, amino-substituted phenyl, carbonyl-substituted phenyl, aldehyde-substituted phenyl, mercapto-substituted phenyl, heterocyclic-substituted phenyl, alkyl-substituted styryl, alkoxy-substituted styryl, halogen-substituted styryl, nitro-substituted styryl, cyano-substituted styryl, trifluoromethyl-substituted styryl, hydroxy-substituted styryl, amino-substituted styryl, carbonyl-substituted styryl, aldehyde-substituted styryl, mercapto-substituted styryl, heterocyclic-substituted styryl, substituted heterocycle, substituted naphthyl, aliphatic amine (primary amine, secondary amine), cyclic amine, and aniline.

[0011] R2 is selected from monosubstituted, disubstituted, or polysubstituted alkyl-substituted cycloalkyl, alkyl-substituted phenyl, alkoxy-substituted phenyl, halogen-substituted phenyl, nitro-substituted phenyl, cyano-substituted phenyl, trifluoromethyl-substituted phenyl, hydroxy-substituted phenyl, amino-substituted phenyl, carbonyl-substituted phenyl, aldehyde-substituted phenyl, mercapto-substituted phenyl, heterocyclic-substituted phenyl, substituted heterocycle, and substituted naphthyl.

[0012] The pharmaceutically acceptable salts of the compounds in the above general formula are hydrochloride, sulfate, hydrogen sulfate, hydrobromide, oxalate, citrate, methanesulfonate, etc.

[0013] The present invention also provides a method for preparing the above-mentioned compounds and their salts.

[0014] Synthesis method of general formula (1):

[0015]

[0016] The synthesis of the salts of the compounds of this invention is based on the above reactions, with the following further reactions:

[0017]

[0018] The specific compounds involved in the above preparation method are prepared as follows:

[0019] Preparation of compound (II)

[0020] Sophoridine (I) reacts with Lawson's reagent in toluene to form compound (II).

[0021] Preparation of compound (III)

[0022] Compound (II) reacts with nitromethane and 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) to form compound (III).

[0023] Preparation of compound (IV)

[0024] Compound (III) was refluxed with zinc powder in acetic acid to form compound (IV).

[0025] Preparation of general formula (1)

[0026] Compound (IV) was reacted with N,N-diisopropylcarbodiimide (DIC), 1-hydroxybenzotriazole (HOBt) and substituted carboxylic acids in dichloromethane to give general formula (1).

[0027] Preparation of general formula (3)

[0028] Preparation of compound salts

[0029] General formula (1) reacts with acids to obtain their corresponding salts, where HA is hydrochloric acid, sulfuric acid, hydrogen sulfate, hydrobromic acid, oxalic acid, citric acid, methanesulfonic acid, etc.

[0030] Synthesis method of general formula (2):

[0031]

[0032] The synthesis of the salts of the compounds of this invention is based on the above reactions, with the following further reactions:

[0033]

[0034] The specific compounds involved in the above preparation method are prepared as follows:

[0035] Preparation of general formula (2)

[0036] Compound (IV) was reacted with triethylamine and substituted sulfonyl chloride in dichloromethane to give general formula (2).

[0037] Preparation of general formula (4)

[0038] Preparation of compound salts

[0039] General formula (2) reacts with acids to obtain their corresponding salts, where HA is hydrochloric acid, sulfuric acid, hydrogen sulfate, hydrobromic acid, oxalic acid, citric acid, methanesulfonic acid, etc.

[0040] The chemical structures, yields, and mass spectrometry data of some preferred compounds synthesized in this invention are shown in Table 1.

[0041] Table 1 shows the structure, yield, mass spectrum, and molecular formula of some preferred compounds.

[0042]

[0043]

[0044]

[0045] Note: The measured values ​​of C, H, and N differ from the theoretical calculations by 0.3%.

[0046] NMR data of some preferred compounds:

[0047] A: 1 H NMR (600MHz, CDCl3) δ7.57(dd,J=8.0,0.9Hz,1H),7.47(dd,J=7.6,1.6Hz,1H),7.34(td,J=7.5,1.0Hz,1H),7.28–7.2 5(m,2H),5.71(dd,J=13.5,4.5Hz,1H),4.65–4.58(m,1H),3.59(dd,J=20.0,8.6Hz,2H),3.56–3.46(m,2H),3.22(s,1H ),3.12(dd,J=18.4,5.1Hz,1H),3.08–3.04(m,1H),2.95(dd,J=18.3,6.2Hz,1H),2.71(dd,J=16.9,8.8Hz,2H),2.47(s ,1H),2.25(d,J=8.4Hz,1H),2.21–2.16(m,1H),2.11(q,J=12.2Hz,3H),1.96–1.88(m,2H),1.83(ddd,J=13.7,6.6,3.0 Hz,1H),1.80–1.75(m,2H),1.72(d,J=15.3 Hz,1H),1.63(ddd,J=19.0,10.2,4.4 Hz,1H). 13 C NMR(151 MHz, CDCl3)δ198.81,168.76,137.61,133.32,131.36,129.30,127.66,119.22,64.95,56.40 ,56.22,53.85,48.78,45.61,42.58,41.31,34.00,28.18,28.05,25.66,24.52,18.99,18.54.

[0048] B: 1H NMR(600 MHz,CDCl3)δ8.59(s,1H),8.28–8.25(m,2H),8.20–8.18(m,2H),5.83(dd,J=13.8,4.4 Hz,1H),4.84(dd,J=16.8,10.1 Hz,1H),3.70(dt,J=13.7,5.7Hz,1H),3.56(dd,J=26.9,11.8 Hz,2H),3.46(t,J=13.5 Hz,1H),3.24–3.18(m,2H),2.97(dd,J=18.4,5.0Hz,1H),2.88(ddd,J=13.9,11.5,6.0 Hz,1H),2.78(s,2H),2.61(s,1H),2.31–2.26(m,2H),2.20(dd,J=22.6,10.4 Hz,2H),2.09(d,J=14.2Hz,1H),1.97(d,J=14.0 Hz,1H),1.85(ddd,J=16.1,11.5,5.0 Hz,3H),1.74(d,J=14.8 Hz,1H),1.67–1.58(m,2H). 13 C NMR(151 MHz,CDCl3)δ199.43,166.28,149.73,139.20,128.77,123.69,64.94,56.63,56.54,52.78,48.26,45.29,42.27,41.52,33.77,28.42,27.92,25.58,24.32,18.98,18.52.

[0049] C: 1 H NMR(600 MHz,CDCl3)δ8.00(t,J=5.5 Hz,1H),7.84(d,J

[0050] =8.2 Hz,2H),7.23(d,J=8.2 Hz,2H),5.77(dd,J=13.7,4.4 Hz,1H),4.75(dd,J=18.1,7.5 Hz,1H),3.60–3.54(m,3H),3.50(t,J=13.5 Hz,1H),3.21(s,1H),3.08–2.93(m,3H),2.75(t,J=12.4 Hz,2H),2.65–2.62(m,2H),2.54(d,J=4.4 Hz,1H),2.22(dd,J=30.6,14.0 Hz,2H),2.18–2.10(m,2H),2.05(d,J=14.9 Hz,1H),1.91(t,J=16.3 Hz,2H),1.84–1.77(m,2H),1.73–1.64(m,2H),1.60(dq,J=15.3,7.7 Hz,3H),1.38–1.30(m,2H),0.92(t,J=7.4 Hz,3H). 13 C NMR(151 MHz,CDCl3)δ199.22,168.75,147.33,130.75,128.66,127.36,64.91,56.52,56.38,53.32,48.50,45.46,41.99,41.78,35.56,33.89,33.31,28.17,27.98,25.61,24.35,22.33,18.99,18.51,13.91.

[0051] D: 1 H NMR(600 MHz,CDCl3)δ8.02(t,J=5.5 Hz,1H),7.88(d,J

[0052] =8.4 Hz,2H),7.45(d,J=8.4 Hz,2H),5.79(dd,J=13.7,4.4 Hz,1H),4.76(dd,J=18.4,7.7 Hz,1H),3.61–3.55(m,3H),3.50(t,J=13.4 Hz,1H),3.21(s,1H),3.08(d,J=18.8 Hz,1H),3.01(dd,J=18.4,4.9 Hz,1H),2.93(ddd,J=13.8,10.5,6.1 Hz,1H),2.74(t,J=12.6 Hz,2H),2.56(dd,J=9.6,4.8 Hz,1H),2.28–2.13(m,4H),2.05(d,J=14.7 Hz,1H),1.91(dd,J=26.8,12.8 Hz,2H),1.80(ddd,J=14.0,9.4,4.3 Hz,2H),1.71(d,J=15.1 Hz,1H),1.66–1.57(m,2H),1.32(s,9H). 13 CNMR(151 MHz,CDCl3)δ199.32,168.54,155.37,130.56,127.20,125.56,64.93,56.53,56.40,53.25,48.47,45.45,41.89,41.85,34.98,33.88,31.18,28.18,27.95,25.63,24.38,18.99,18.53.

[0053] E: 1 H NMR(600 MHz,CDCl3)δ8.38(t,J=5.5 Hz,1H),8.08(d,J

[0054] =8.2 Hz,2H),7.69(d,J=8.3 Hz,2H),5.80(dd,J=13.7,4.3 Hz,1H),4.77(d,J=7.5 Hz,1H),3.63(dd,J=13.5,6.2 Hz,1H),3.57(dd,J=22.5,11.6 Hz,2H),3.46(t,J=13.5 Hz,1H),3.23(s,1H),3.14(d,J=18.5 Hz,1H),3.03–2.93(m,2H),2.78(s,2H),2.57(d,J=4.7 Hz,1H),2.26(dd,J=23.0,12.2 Hz,2H),2.17(dd,J=22.8,9.4 Hz,2H),2.07(d,J=14.7 Hz,1H),1.95(d,J=14.5 Hz,1H),1.91(d,J=11.5 Hz,1H),1.87–1.80(m,2H),1.74(d,J=15.1 Hz,1H),1.67–1.61(m,2H). 13 CNMR(151 MHz,CDCl3)δ199.28,167.55,160.86(d,J=38.6 Hz),136.60,133.49(d,J=32.4 Hz),127.95,125.63,123.74(d,J=272.5 Hz),115.72(d,J=289.0 Hz),65.00,56.63,56.51,53.03,48.36,45.30,42.03,41.85,33.82,28.29,27.95,25.57,24.30,18.97,18.50.

[0055] F: 1 H NMR(600 MHz,CDCl3)δ7.71(d,J=15.9 Hz,1H),7.67(s,

[0056] 1H),7.51(t,J=7.0 Hz,1H),7.30(td,J=7.3,1.5 Hz,1H),7.13(t,J=7.2 Hz,1H),7.08–7.03(m,1H),6.65(d,J=15.9 Hz,1H),5.74(dd,J=13.7,4.4 Hz,1H),4.65–4.60(m,1H),3.59(t,J=12.9 Hz,2H),3.54–3.46(m,2H),3.24(s,1H),3.05–2.92(m,3H),2.76(dd,J=17.4,11.9Hz,2H),2.44(s,1H),2.26(d,J=11.8 Hz,1H),2.21–2.05(m,4H),1.91(d,J=12.0 Hz,2H),1.83–1.69(m,4H),1.66–1.60(m,1H). 13 CNMR(151 MHz,CDCl3)δ199.08,167.16,161.49(d,J=251.6 Hz),133.93,131.15(d,J=8.7 Hz),129.16,124.44(d,J=3.5 Hz),123.14,122.83(d,J=11.6 Hz),116.09(d,J=21.9 Hz),64.89,56.44,56.34,53.53,48.61,45.48,41.80,41.62,33.90,28.21,28.10,25.59,24.35,18.99,18.52.

[0057] G: 1 H NMR(600 MHz,CDCl3)δ7.84(s,1H),7.55(d,J=15.8 Hz,

[0058] 1H),7.33–7.26(m,2H),7.17(t,J=8.0 Hz,1H),7.02(t,J=8.3 Hz,1H),6.54(t,J=10.3 Hz,1H),5.72(dt,J=22.1,11.1 Hz,1H),4.63–4.53(m,1H),3.57(t,J=12.2 Hz,2H),3.47(dd,J=24.1,10.8 Hz,2H),3.25(s,1H),2.99(t,J=12.2 Hz,3H),2.78(d,J=8.3 Hz,2H),2.39(s,1H),2.25(d,J=13.0 Hz,1H),2.17–2.05(m,4H),1.89(d,J=12.5Hz,2H),1.81(d,J=14.8 Hz,2H),1.76–1.68(m,2H),1.64(dd,J=23.8,9.4 Hz,1H). 13 CNMR(151 MHz,CDCl3)δ198.84,167.37,162.97(d,J=246.4 Hz),140.46,136.86(d,J=7.7Hz),130.45(d,J=8.2 Hz),124.00,121.38,116.79(d,J=21.3 Hz),114.11(d,J=21.9Hz),64.81,56.41,56.29,53.65,48.60,45.39,41.98,41.36,33.85,28.27,28.06,25.43,24.13,18.93,18.45.

[0059] H: 1 H NMR(600 MHz,CDCl3)δ7.71(t,J=6.0 Hz,1H),7.58(d,J

[0060] =15.8 Hz,1H),7.51–7.47(m,2H),7.04(t,J=8.6 Hz,2H),6.49–6.44(m,1H),5.77(dd,J=13.7,4.4 Hz,1H),4.65(dt,J=11.2,8.1 Hz,1H),3.63–3.56(m,2H),3.52(dd,J=13.3,6.9 Hz,1H),3.47(d,J=13.5 Hz,1H),3.22(s,1H),3.09–3.03(m,1H),2.99(dd,J=18.3,5.0 Hz,1H),2.87(ddd,J=14.0,10.1,6.2 Hz,1H),2.75(ddd,J=14.6,9.8,2.8 Hz,2H),2.47(dd,J=9.4,4.6 Hz,1H),2.30–2.20(m,2H),2.17–2.07(m,3H),1.91(t,J=16.0 Hz,2H),1.80(ddd,J=14.2,9.8,4.9 Hz,2H),1.75(d,J=15.0 Hz,1H),1.69–1.59(m,2H). 13 C NMR(151 MHz,CDCl3)δ199.31,167.40,163.62(d,J=250.4 Hz),139.93,131.08,129.74(d,J=8.3 Hz),120.24,115.92(d,J=21.9 Hz),64.93,56.51,56.42,53.26,48.46,45.40,41.89,41.52,33.86,28.31,28.04,25.61,24.37,18.99,18.54.

[0061] J: 1 H NMR(600MHz,CDCl3)δ7.61(dd,J=3.7,1.2Hz,1H),7.58

[0062] (dd,J=5.0,1.1Hz,1H),7.10(dd,J=4.9,3.8Hz,1H),6.66(t,J=6.2Hz,1H),5.70(dd,J=13.6,4.4Hz,1H),4.6 5–4.59(m,1H),3.57(dd,J=17.8,12.6Hz,2H),3.46(t,J=13.5Hz,1H),3.26(s,1H),3.03(dd,J=18.4,4.9Hz, 1H),2.95(ddd,J=30.1,15.7,12.2Hz,2H),2.78(dd,J=26.2,12.9Hz,2H),2.71–2.66(m,1H),2.35–2.16(m,4 H),2.10(dd,J=25.3,8.9Hz,2H),1.94–1.87(m,2H),1.83–1.73(m,4H),1.67(ddd,J=14.4,10.4,4.1Hz,1H). 13 C NMR (151MHz, CDCl3) δ198.66,140.75,131.88,131.66,127.55,64.59,56.25,53. 40,48.50,45.19,44.49,41.43,33.84,29.71,27.89,25.54,24.36,18.93,18.53.

[0063] Bioactivity tests showed that these compounds could inhibit the expression of NO, TNF-α and IL-6 in mouse RAW 246.7 cells after LPS stimulation, and could be used to prepare related drugs for the treatment of inflammation.

[0064] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0065] This invention provides novel matrine compounds and their chemical structures, offering new compound resources for researchers in the field of pharmaceutical technology. It can serve as a reference for innovation and new drug development in the pharmaceutical field, particularly for drugs related to the treatment of inflammation. Attached Figure Description

[0066] Figure 1 It is compound A in the embodiments of the present invention. 1 H NMR spectrum;

[0067] Figure 2 It is compound A in the embodiments of the present invention. 13 C NMR spectrum;

[0068] Figure 3 It is compound B in the embodiments of the present invention. 1 H NMR spectrum;

[0069] Figure 4 It is compound B in the embodiments of the present invention. 13 C NMR spectrum;

[0070] Figure 5 It is compound C in the embodiments of the present invention. 1 H NMR spectrum;

[0071] Figure 6 It is compound C in the embodiments of the present invention. 13 C NMR spectrum;

[0072] Figure 7 It is compound D in the embodiments of the present invention. 1 H NMR spectrum;

[0073] Figure 8 It is compound D in the embodiments of the present invention. 13 C NMR spectrum;

[0074] Figure 9 It is compound E in the embodiments of the present invention. 1 H NMR spectrum;

[0075] Figure 10 It is compound E in the embodiments of the present invention. 13 C NMR spectrum;

[0076] Figure 11 It is compound F in the embodiments of the present invention. 1 H NMR spectrum;

[0077] Figure 12 It is compound F in the embodiments of the present invention. 13 C NMR spectrum;

[0078] Figure 13 It is compound G in the embodiments of the present invention. 1 H NMR spectrum;

[0079] Figure 14 It is compound G in the embodiments of the present invention. 13 C NMR spectrum;

[0080] Figure 15 It is compound H in the embodiments of the present invention. 1 H NMR spectrum;

[0081] Figure 16 It is compound H in the embodiments of the present invention. 13 C NMR spectrum;

[0082] Figure 17 It is compound J in the embodiments of the present invention. 1H NMR spectrum;

[0083] Figure 18 It is compound J in the embodiments of the present invention. 13 C NMR spectrum. Detailed Implementation

[0084] To enable those skilled in the art to better understand the present invention, the technical solution of the present invention will be described in further detail below with reference to the embodiments and accompanying drawings. Obviously, the described embodiments are merely some, not all, of the embodiments of the present invention. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative effort should fall within the scope of protection of the present invention.

[0085] It should be noted that, unless otherwise specified, the embodiments and features described in the present invention can be combined with each other. The present invention will now be described in detail with reference to the embodiments.

[0086] Example 1: Preparation of Thiosophoridine

[0087] 50.00 g (203.11 mmol, purchased from Ningxia Bauhinia Pharmaceutical Co., Ltd.) of sophoridine was placed in a 1 L round-bottom flask, and 500 mL of toluene and 45.18 g (111.71 mmol) of Lawson's reagent were added. The reaction was detected by TCL. After about 2 hours, the reaction was completed and the reaction was stopped. The mixture was filtered while hot, the solvent was evaporated to dryness, and the mixture was separated by silica gel column chromatography (CH2Cl2 / CH3OH = 50 / 1) to give 30.30 g of compound II as a yellow oil, with a yield of 56.9%.

[0088] Example 2: Preparation of 13-methylenenitrothiomatrine (Compound III)

[0089] 30.30 g (115.46 mmol) of compound II, 422.86 g (6.93 mol) of nitromethane, and 7.03 g (46.18 mmol) of 1,8-diazabicyclo[5.4.0]undec-7-ene were placed in a 500 mL round-bottom flask and stirred overnight at room temperature. The reaction was complete as detected by TCL. The reaction mixture was evaporated to dryness and separated by silica gel column chromatography (CH2Cl2 / CH3OH = 50 / 1) to give 34.01 g of compound III as a yellow solid, with a yield of 91.1%.

[0090] Example 3: Preparation of 13-methyleneaminothiomatrine (compound IV)

[0091] Compound III (30.00 g, 92.83 mmol), zinc powder (24.58 g, 371.32 mmol), and acetic acid (250 mL) were placed in a 500 mL reaction flask and stirred overnight at room temperature. The reaction was detected by TCL and was considered complete. The mixture was filtered through diatomaceous earth, evaporated to dryness, and separated by silica gel column chromatography (CH2Cl2 / CH3OH = 20 / 1) to give compound IV (20.76 g, a yellow oily substance, yield 76.3%).

[0092] Example 4: Preparation of compound A in the table

[0093] Compound IV 120.03 mg (0.409 mmol), DIC 154.85 mg (1.227 mmol), HOBt 165.79 mg (0.227 mmol), 2-bromobenzoic acid 246.65 mg (1.227 mmol), and 10 mL of dichloromethane were placed in a 50 mL reaction flask and stirred overnight at room temperature. The reaction was monitored by TCL, and the reaction was considered complete. The reaction solution was evaporated to dryness, and the mixture was separated by silica gel column chromatography (CH2Cl2 / CH3OH = 50 / 1) to give compound A1 10.60 g. It was a white solid, with a yield of 53.4%.

[0094] Example 5: Preparation of compound F in the table

[0095] Compound IV (120.03 mg, 0.409 mmol), DIC (154.85 mg, 1.227 mmol), HOBt (165.79 mg, 0.227 mmol), 2-fluorocinnamic acid (203.83 mg, 1.227 mmol), and 10 mL of dichloromethane were placed in a 50 mL reaction flask and stirred overnight at room temperature. The reaction was monitored by TCL, and the reaction was considered complete. The reaction solution was evaporated to dryness, and the mixture was separated by silica gel column chromatography (CH2Cl2 / CH3OH = 50 / 1) to give compound F (101.61 mg). The result was a pale yellow oil with a yield of 56.27%.

[0096] Example 6: Preparation of compound J in the table

[0097] Compound IV 120.03 mg (0.409 mmol) and 10 mL of dichloromethane were placed in a 50 mL reaction flask. At 0 °C, triethylamine 124.16 mg (1.227 mmol) and thiophene sulfonyl chloride 224.11 mg (1.227 mmol) were added, and the mixture was transferred to room temperature and stirred overnight. The reaction was complete as detected by TCL. The reaction solution was evaporated to dryness, and the mixture was separated by silica gel column chromatography (CH2Cl2 / CH3OH = 50 / 1) to give compound J 126.24 mg. It was a white solid, with a yield of 57.3%.

[0098] Example 7: Preparation of 13-methyleneamino-(o-bromobenzamide)thiomatrine hydrochloride

[0099] 100.00 mg (0.21 mmol) of 13-methyleneamino-(o-bromobenzamide)thiomatrine (compound A) was dissolved in 20 mL of diethyl ether, and 1 mL of hydrochloric acid was added. The mixture was stirred at room temperature for 2 h. The reaction was confirmed to be complete by TCL detection. The reaction solution was concentrated under reduced pressure, and the precipitate was filtered to obtain 98.00 mg of compound A hydrochloride, with a yield of 91.0%. Other compound hydrochloride salts can be obtained using the same method.

[0100] The implementation of this invention is not limited to the above embodiments. All reagents used in the embodiments are commercially available analytical grade reagents.

[0101] Example 8: Bioactivity Test

[0102] 8.1 Instruments and Equipment

[0103] Inverted biological microscope, CO2 cell incubator, vertical single-sided double-person clean bench, enzyme-linked immunosorbent assay (ELISA) analyzer.

[0104] 8.2 Experimental Materials and Preparation

[0105] Test drug:

[0106] Compounds AH, J, and M19 (structural formulas shown below)

[0107]

[0108] Appearance: Powder and oily substance

[0109] Content: 99%

[0110] Preparation method: Dissolve in DMSO and dilute with PBS to the required concentration of the sample;

[0111] Cell lines:

[0112] RAW264.7 (mouse macrophages) were purchased from the Shanghai Institute of Cell Biology, Chinese Academy of Sciences. 8.2.3 Reagents:

[0113] Cell culture medium: DMEM (Corning), fetal bovine serum (Gibco), penicillin / streptomycin (Gibco).

[0114] Phosphate-buffered saline (PBS): KCl 0.2g, NaCl 8.0g, KH2PO4 0.27g, Na2HPO4·12H2O 3.5814g, dissolved in 1L double-distilled water, autoclaved at 120℃ for 30min, and stored at 4℃.

[0115] Sample solvent: DMSO (Sigma).

[0116] Bioactivity assay kits: CCK8 kit (Beyotime), nitric oxide assay kit (Beyotime), ELISA kit (Elabscience).

[0117] Experimental consumables: 96-well cell culture plates (Corning), cell culture dishes (Corning), centrifuge tubes (Sinopharm Group).

[0118] 8.3 Experimental Procedure

[0119] Cytotoxicity test

[0120] When cells are in the logarithmic growth phase, at a rate of 2 × 10 4 Cells were seeded per well in 96-well plates and treated with matrine derivatives (3.125 μM, 6.25 μM, 12.5 μM, 25 μM, and 50 μM) for 24 h. The culture medium was then discarded, and a prepared CCK8 solution was added. The plates were incubated in a CO2 incubator for 2 h. The absorbance at 450 nm was measured to calculate cell viability.

[0121] NO inhibition rate test

[0122] When cells are in the logarithmic growth phase, at a rate of 2 × 10 4 Each well was seeded with matrine derivative (12.5 μM) for 4 h, then the culture medium was discarded, and the cells were treated with LPS (100 ng / mL) and the compound (12.5 μM) for 14 h. The relative NO content was determined using a nitric oxide assay kit. The absorbance at 540 nm was measured.

[0123] ELISA test

[0124] When cells are in the logarithmic growth phase, at a rate of 2 × 10 4 Each well was seeded with 1 sample of matrine derivative (12.5 μM) for 4 h. After pretreatment, the culture medium was discarded, and the plates were treated with LPS (100 ng / mL) and matrine derivative (12.5 μM) for 14 h. The levels of TNF-α and IL-6 were detected using an ELISA kit.

[0125] 8.4 Experimental Results:

[0126] Cytotoxicity test results

[0127] Table 2. Results of cytotoxicity assays of matrine derivatives on RAW 264.7 cells.

[0128]

[0129] Results of NO, TNF-α and IL-6 inhibition rate assay

[0130] Table 3. Effects of matrine derivatives at 12.5 μM on LPS-induced release of NO, TNF-α, and IL-6 in RAW 264.7 cells.

[0131]

[0132] Note: (*p<0.05,**p<0.01,***p<0.001 versus M19)

[0133] Table 3 shows that the novel thiomatrine derivative can inhibit the expression of NO, TNF-α and IL-6 in mouse RAW 246.7 cells after LPS stimulation, and the inhibition rate is significantly higher than that of M-19. It can be used to prepare related candidate drugs for inflammation treatment.

[0134] The above specific embodiments are merely explanations of the present invention and are not intended to limit the present invention. After reading this specification, those skilled in the art can make modifications to these embodiments without contributing any inventive step, but as long as they are within the scope of the claims of the present invention, they are protected by patent law.

Claims

1. Matrine compounds and their pharmaceutically acceptable salts, whose general structural formulas are shown in formula (1) or formula (2) below: The matrine-like compounds represented by general formula (1) are one of the compounds shown in the following structures: 、 、 、 、 、 、 、 ； The matrine compounds represented by general formula (2) are compounds with the following structures: 。 2. The method for preparing the matrine compounds and their pharmaceutically acceptable salts as described in claim 1, characterized in that: React according to the following route: The synthesis of compound salts is based on the above reactions, further performed by the following reactions: HA can be hydrochloric acid, sulfuric acid, hydrobromic acid, oxalic acid, citric acid, or methanesulfonic acid.

3. The use of the matrine compounds of claim 1 and their pharmaceutically acceptable salts in the preparation of anti-inflammatory drugs.

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