CN103936736A - Matrine derivative and preparation method thereof - Google Patents

Abstract

The present invention relates to a class of matrine derivatives and their salts, in particular to a new class of matrine compounds and their salts having the following general chemical structure: In the general formula, X is a sulfur atom; n is selected from 1,2; _R is selected from alkyl, alkoxy, aryl, substituted aryl; _R is selected from hydrogen, alkyl, substituted alkyl; the present invention also Provided are a preparation method of a matrine derivative and a salt thereof. The preparation method of the invention has high yield, the compound has good antitumor effect, and has the advantages of targeting, low toxicity, and the like; the target molecule obtained by the preparation method of the invention has high stability. The activity of some compounds is obviously better than that of compound M19.

Description

A kind of matrine derivative and its preparation method

technical field

The invention relates to the technical field of medicine, in particular to a class of matrine derivatives and a preparation method thereof.

Background technique

Liver fibrosis is the common pathological basis of all chronic liver diseases. It is caused by abnormal proliferation of connective tissue in the liver caused by various pathogenic factors, leading to excessive precipitation of diffuse extracellular matrix (ECM) in the liver. Pathological process [Kotaro S et al, Biochem Biophys Res Commun , 2013, 443(3): 950-956]. If the liver fibrosis develops further, resulting in the reconstruction of liver lobule, the formation of pseudolobule and nodule, it will become substantial liver cirrhosis, which is difficult to reverse, so it is particularly important to control the occurrence and development of liver fibrosis. Studies on the mechanism of liver fibrosis have shown that the activation of hepatic stellate cells (HSC) is the central link in the occurrence of liver fibrosis [Hu L et al , Drug Discov Ther, 2013, 7(6): 248- 53.], under the action of pathological factors, hepatic parenchymal cells are damaged, which in turn stimulates the production and release of growth factors that can promote HSCs activation [Borg B et al, Liver Transp , 201l, l7(7): 814-823.]. These factors pass through intracellular signaling pathways, such as TGF-β/Smad pathway [Sancho-Bru P et al, Liver Int , 2010, 30(1): 31- 41.], Ras/ERK [Chen X et al, Genet Mol Res , 2013, 12(1): 665-77.] Pathways, etc., transmit information to the nucleus, produce corresponding biological effects, and lead to HSC proliferation. And after HSC is activated, it transforms into myofibroblasts, expresses α-smooth muscle actin (α-SMA) [Gressner A, Kidney Int Suppl, 1996, 54: S39-45.], and secretes A large number of ECM, including collagen (Collagen I, Collagen III), fibronectin (fibronectin) and proteoglycan, promote the process of liver fibrosis. Therefore, the research on inhibiting the activation of HSC is the main direction of research on the treatment of liver fibrosis.

In the process of liver fibrosis, the current research is relatively clear. In addition to the above-mentioned EGFR, EGF and VEGF, there are some factors and signaling pathways involved in the occurrence and development of liver fibrosis: matrix metalloproteinase (MMP) is a group that can degrade All ECM enzymes except polysaccharides, and matrix metalloproteinase inhibitor (TIMP) is a specific inhibitor of MMP, which can specifically bind to the active center of MMPs to inhibit its activity. The most active in the liver is TIMP-1[ Rockey D, Hepatology , 1999, 30(3): 816-8.]. The two work together to regulate the dynamic balance of the degradation and deposition of ECM in the liver. When liver fibrosis occurs, the expression of the two will also change.

Matrine and Oxymatrine are the active ingredients of S. flavescens Ait (Figure 1). It has been found that matrine has anti-inflammatory, immunosuppressive, anti-tumor and anti-hepatic fibrosis pharmacological effects [ Liu Y et al, 2014, doi: 10.1007/s13277-014-1680-z. ], which has been clinically used in acute Treatment of chronic liver disease. Studies have shown that matrine and oxymatrine have anti-hepatic fibrosis effects in rats [Zhang J et al, Acta Pharmacol. Sin , 2001, 22(2), 183-186.], but the pharmacological effects of matrine are not Strong, oxymatrine has a short half-life, t1/2 is only 0.5h, and the drug is eliminated quickly. Therefore, in cooperation with the School of Pharmacy of the Second Military Medical University, the structure is modified in the hope of finding compounds with higher pharmacological activity. Our research group has confirmed that the carbonyl group is the pharmacophore of matrine through previous studies. Through thioxo and side chain Michael addition, a series of thiomatrine compounds were synthesized, and their pharmacological activities were compared. M19 was found (Fig. 1) It is one of the most active compounds, and its toxicity is comparable to that of matrine [Wu Qiuye et al., Matrine Compounds and Their Preparation and Application, CN101704817A]. Zhang Junping’s research found that M19 has anti-hepatic fibrosis and other diseases [Zhang Junping, 13-methylamino-18-thiomatrine compound used in the preparation of anti-hepatic fibrosis or other tissue and organ fibrosis drugs, CN 102258516 A], follow-up research found that matrine derivative M-19 has good activity, but its stability is poor, and it is easy to be eliminated when heated, which affects its application in the preparation of anti-hepatic fibrosis drugs. This application is based on the previous research. Its aminoacylation improves the stability of the target molecule. The activity of some compounds is obviously better than that of compound M19.

Contents of the invention

The object of the present invention is to provide a class of matrine derivatives and salts thereof to address the deficiencies in the prior art.

Another object of the present invention is to provide a preparation method of matrine derivatives and salts thereof.

For realizing above-mentioned object, the technical scheme that the present invention takes is:

A class of matrine derivatives and salts thereof, the structure of which is shown in the general formula:

                                                 

Wherein X is a sulfur atom;

wherein n is selected from 1 or 2;

Wherein R ₁ , R ₂ are selected from or or :

． Alkyl group selected from methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy or benzyloxy, preferably methoxy;

． Aryl, selected from benzyl, p-fluorobenzyl, p-chlorobenzyl, o-fluorobenzyl, p-methylbenzyl, preferably p-methylbenzyl;

． Cyclic amino group, cyclic amino group is piperidinyl, tetrahydropyrrolyl and piperazinyl.

Said X is a sulfur atom.

Said X is a sulfur atom, and n is 1 or 2.

The matrine derivative is 13-[ N -methyl, N- (2-allylaminoacetyl)]-amino-15-thiomatrine.

The matrine derivative is 13-[ N -methyl, N- (4-fluorophenethylaminoacetyl)]-amino-15-thiomatrine.

Said salt is hydrochloride, sulfate, hydrogensulfate, hydrobromide, oxalate, citrate or methanesulfonate.

For realizing above-mentioned second purpose, the technical scheme that the present invention takes is:

The preparation method of the described matrine derivative and salt thereof, the method is as follows;

The synthetic route of WM-1 series compounds:

Synthetic routes of WM-2 series compounds:

 

Synthetic routes of WM-3 series compounds:

The synthesis of compound salts of the present invention is on the basis of above-mentioned reaction, further reacts as follows:

HA is hydrochloric acid, sulfuric acid, hydrogen sulfate, hydrobromic acid, oxalic acid, citric acid or methanesulfonic acid.

The advantages of the present invention are that: the present invention provides a new class of matrine derivatives for the preparation of anti-hepatic fibrosis and anti-liver cancer drugs, the preparation method of the present invention has high yield, the compound has good anti-tumor effect, and has targeting, low toxicity, etc.; the target molecule obtained by the preparation method of the present invention has high stability. The activity of some compounds is obviously better than that of compound M19.

Description of drawings

Figure 1. Chemical structure of matrine, M19.

Figure 2. The effect of matrine derivatives on the proliferation of liver cancer cell line Hep-G2.

Figure 3. Effects of matrine derivatives on the proliferation of liver cancer cell line Hun-7.

Figure 4. The effect of matrine derivatives on the proliferation of liver cancer cell line MHCC-97L.

Figure 5. The effect of matrine derivatives on the proliferation of liver cancer cell line SMMC-7721.

Figure 6. Effects of matrine derivatives on the proliferation of liver cancer cell line Hep-3B.

Figure 7. Effects of matrine derivatives on BJ proliferation of fibroblasts.

Figure 8. Effects of matrine derivatives on the proliferation of human hepatic stellate cells LX-2.

Figure 9. Effects of matrine derivatives on the proliferation of rat hepatic stellate cells HSC-T6.

Figure 10. The effect of WM-108 on inhibiting Hep3B. As the concentration increases, the inhibition rate of WM-108 on Hep3B also gradually increases.

Figure 11. WM-108 inhibits the migration and invasion of Hep3B. As the concentration increases, the migration and invasion of cells are significantly inhibited.

Figure 12. The cell morphology of LX-2 cells treated with different concentrations of WM130 for 24 hours. As the concentration increased, the cell proliferation slowed down, and the cell shape became smaller and rounder.

Detailed ways

The specific embodiments provided by the present invention will be described in detail below in conjunction with the accompanying drawings.

Embodiment 1: Preparation of 15-thiosophocarpine

1.0 g (0.004 mol) of sophocarpine and 2.0 g (0.005 mol, purchased from Alfa Company) of sophocarpine and 2.0 g of Lawson's reagent were placed in a 50 ml reaction bottle, 20 ml of dichloromethane was added, and the reaction was heated under reflux for 12 hours. After the reaction was completed, the solvent was removed by concentration under reduced pressure, and the crude product was passed through a silica gel column with dichloromethane:methanol (25:1) as the eluent to obtain 0.96 g of the product, with a yield of 91.2%.

Example 2: Preparation of 13-( N -methyl)-amino-15-thiomatrine (M19)

Put 100 mg (0.0004 mol) of 18-thiosophocarpine in a 20 ml reaction bottle, add 2 ml of methylamino alcohol solution and 1 ml of triethylamine, and stir for 12 hours. After the reaction was completed, the solvent was removed by concentration under reduced pressure, and the crude product was passed through a silica gel column with methylene chloride:methanol (20:1) as the eluent to obtain 98 mg of the product with a yield of 83.5%.

Example 3: Preparation of 13-( N -methyl, N -chloroacetyl)-amino-18-thiomatrine (WM-1 in the table)

M-19 (2 g, 6.8 mmol) and anhydrous potassium carbonate (1 g, 7.2 mmol) were added to a round-bottomed flask, vacuumized and under the protection of an Ar balloon, 40 mL of anhydrous dichloromethane was injected. After ice bathing for 10 min, slowly drop 10 mL of anhydrous dichloromethane diluted chloroacetyl chloride (0.62 mL, 7.9 mmol) into the bottle under low temperature and rapid magnetic stirring. h The reaction was completed and monitored by TLC (dichloromethane/methanol, V/V, 10:1). Filter with celite, concentrate the filtrate under reduced pressure, add methanol to solidify and wash. The solid was dissolved in dichloromethane again, and an equal volume of methanol was added, concentrated under reduced pressure until a large amount of solid precipitated, filtered with suction to obtain a relatively pure product, and dried under reduced pressure to obtain 1.1 g, with a yield of 43.6%.

Example 4: Preparation of 13-[ N -methyl, N- (2-chloropropionyl)]-amino-18-thiomatrine (WM-2 in the table)

Add M-19 (4 g, 13.6 mmol) and anhydrous potassium carbonate (2 g, 14.4 mmol) into a round-bottomed flask, vacuumize and inject 100 mL of anhydrous dichloromethane under the protection of an Ar balloon. After ice bathing for 10 min, slowly drop 2-chloropropionyl chloride (1.62 mL, 16 mmol) diluted with 20 mL of anhydrous dichloromethane into the bottle while maintaining low temperature and rapid magnetic stirring. The reaction was completed in about 1 h and monitored by TLC (dichloromethane/methanol, V/V, 10:1). Filter with celite, concentrate the filtrate under reduced pressure, add methanol to solidify and wash. The solid was then dissolved in dichloromethane, an equal volume of methanol was added, concentrated under reduced pressure until a large amount of solids were precipitated, and filtered with suction to obtain a relatively pure product, which was dried under reduced pressure to obtain 2.69 g, with a yield of 53.4%.

the

Example 5: Preparation of 13-[ N -methyl, N- (chloropropionyl)]-amino-18-thiomatrine (WM-3 in the table)

Add M-19 (4 g, 13.6 mmol) and anhydrous potassium carbonate (2 g, 14.4 mmol) into a round-bottomed flask, vacuumize and inject 100 mL of anhydrous dichloromethane under the protection of an Ar balloon. After ice bathing for 10 min, slowly drop 2-chloropropionyl chloride (1.62 mL, 16 mmol) diluted with 20 mL of anhydrous dichloromethane into the bottle while maintaining low temperature and rapid magnetic stirring. The reaction was completed in about 1 h and monitored by TLC (dichloromethane/methanol, V/V, 10:1). Filter with celite, concentrate the filtrate under reduced pressure, add methanol to solidify and wash. The solid was then dissolved in dichloromethane, an equal volume of methanol was added, concentrated under reduced pressure until a large amount of solid precipitated, filtered with suction to obtain a relatively pure product, and dried under reduced pressure to obtain 3.2 g, with a yield of 63.7%.

Example 6: Preparation of 13-[ N -methyl, N- (2-methylaminoacetyl)]-amino-15-thiomatrine (compound WM-101 in the table)

Take WM-1 (200 mg, 0.54 mmol), anhydrous potassium carbonate (200 mg, 1.45 mmol), catalytic amount of potassium iodide (about 30 mg), add 8 mL of anhydrous acetonitrile, 0.15 ml of diethylamine (1.44 mmol), Heat to 50°C to react, and the reaction is complete in 4-5 hours. Evaporate toluene to dryness, add 30 mL each of water and dichloromethane, shake to dissolve and separate the liquids, and extract the water layer once with 20 mL of dichloromethane. The organic layers were combined and washed once with 25 mL of saturated sodium chloride solution. The organic layer was collected, dried over anhydrous sodium sulfate, and filtered. After concentration under reduced pressure, it was purified by a thin-layer silica gel column with dichloromethane/methanol (+0.5% triethylamine) as the eluent to obtain 184 mg of a white solid with a yield of 86.5%.

Example 7: Preparation of 13-[ N -methyl, N- (2-methylaminoacetyl)]-amino-15-thiomatrine (compound WM-101 in the table)

Take WM-1 (200 mg, 0.54 mmol), anhydrous potassium carbonate (200 mg, 1.45 mmol), catalytic amount of potassium iodide (about 30 mg), add 8 mL of anhydrous acetonitrile, 0.15 ml of diethylamine (1.44 mmol), Heating to 50°C for reaction, methylamine hydrochloride (48 mg 0.55 mmol), 4-5 h for complete reaction. Evaporate toluene to dryness, add 30 mL each of water and dichloromethane, shake to dissolve and separate the liquids, and extract the water layer once with 20 mL of dichloromethane. The organic layers were combined and washed once with 25 mL of saturated sodium chloride solution. The organic layer was collected, dried over anhydrous sodium sulfate, and filtered. After concentration under reduced pressure, it was purified by a thin-layer silica gel column with dichloromethane/methanol (+0.5% triethylamine) as the eluent to obtain 184 mg of a white solid with a yield of 86.5%.

Example 8: Preparation of 13-[ N -methyl, N- (2-allylaminoacetyl)]-amino-15-thiomatrine (compound WM-108 in the table)

Take WM-1 (200 mg, 0.54 mmol), anhydrous potassium carbonate (200 mg, 1.45 mmol), catalytic amount of potassium iodide (about 30 mg), add 8 mL of anhydrous acetonitrile, 0.15 ml of diethylamine (1.44 mmol), Allylamine (0.15 mL, 0.55 mmol) was heated to 50°C to react, and the reaction was complete in 4-5 h. Evaporate toluene to dryness, add 30 mL each of water and dichloromethane, shake to dissolve and separate the liquids, and extract the water layer once with 20 mL of dichloromethane. The organic layers were combined and washed once with 25 mL of saturated sodium chloride solution. The organic layer was collected, dried over anhydrous sodium sulfate, and filtered. After concentration under reduced pressure, it was purified by a thin-layer silica gel column with dichloromethane/methanol (+0.5% triethylamine) as the eluent to obtain 196 mg of white solid with a yield of 87.6%.

Example 9: Preparation of 13-[ N -methyl, N- (4-fluorophenethylaminoacetyl)]-amino-15-thiomatrine (compound WM-130 in the table)

Take WM-1 (200 mg, 0.54 mmol), anhydrous potassium carbonate (200 mg, 1.45 mmol), catalytic amount of potassium iodide (about 30 mg), add 8 mL of anhydrous acetonitrile, 0.15 ml of diethylamine (1.44 mmol), 4-Fluorophenethylamine (0.19 mL, 0.55 mmol), heated to 50°C for 4-5 h to complete the reaction. Evaporate toluene to dryness, add 30 mL each of water and dichloromethane, shake to dissolve and separate the liquids, and extract the water layer once with 20 mL of dichloromethane. The organic layers were combined and washed once with 25 mL of saturated sodium chloride solution. The organic layer was collected, dried over anhydrous sodium sulfate, and filtered. After concentration under reduced pressure, it was purified by a thin-layer silica gel column with dichloromethane/methanol (+0.5% triethylamine) as the eluent to obtain 226 mg of white solid with a yield of 85.7%.

The implementation of the present invention is not limited to the above examples. The rest of the target compounds are synthesized using different alcohols, mercaptans, and substituted amines as raw materials, and the required matrine compounds can be synthesized by repeating the steps in the above examples. All reagents used in the examples are commercially available analytically pure.

Embodiment 10: biological activity test:

Test materials: liver cancer cell lines Hep-G2, Hun-7, MHCC-97L, SMMC-7721 and Hep-3B fibroblast BJ, human hepatic stellate cell LX-2, rat hepatic stellate cell HSC-T6 purchased from Institute of Cellular Science, Chinese Academy of Sciences.

experiment method:

MTT colorimetric method: use liver cancer cell lines Hep-G2, Hun-7, MHCC-97L, SMMC-7721 and Hep-3B fibroblast BJ, human hepatic stellate cell LX-2, rat hepatic stellate cell HSC- T6 is used for in vitro experiment of drug effect, cultured in 10% FBS DMEM medium at 37°C, 5% CO2 incubator, trypsinized and passaged, cells are seeded into 96-well plates, about 5000 cells per well, cells After 8 hours of wall attachment, add various concentrations of WM130 sodium salt solution (1mg/ml), add 20μL of MTT (5g L-1 ) to each well after 24 hours of action, incubate for another 4 hours, discard the supernatant of each well, and add 150μL DMSO, shake gently for 10 min, and detect the absorbance value with a microplate reader at a wavelength of 492 nm. Determine the half maximal inhibitory concentration (IC50) of WM130 sodium salt on each cell line, and compare the effects. Select several concentration groups with drug efficacy, and set up a blank group in addition, and carry out the following experiments:

Transwell method to detect cell migration: apply a layer of fibronectin (10 ug/ml, 50ul) on the lower surface of the PVPF membrane of Transwell, wash it with PBS at 37°C for 2 hours, and put in the culture medium (containing 10% serum) in a 24-well plate, and then add cells (100ul, dilute different concentrations of drugs with medium containing 0.1% serum) in the inner chamber of Transwell, including the control group, put them in the incubator, after 12-18 hours , take out the Transwell, wipe off the cells on the side of the PVPF membrane close to the inner chamber with a cotton swab, fix the cells on the other side with formaldehyde at room temperature for 30 minutes, stain with crystal violet for 20 minutes, wash with water for more than 3 times, and then observe the cells under a microscope and count them.

Flow cytometry detection of cell apoptosis: cells treated with different concentrations of drugs, including the control group, were washed twice with cold PBS, and then made into a suspension of 1×106 cells/ml with 1×Binding Buffer, and 100 μl of the cell suspension was taken , add Annexin V and mix well with the nucleic acid dye, place at room temperature in the dark for 15 minutes, add 400 μl of 1×Binding Buffer to each group, and measure the results by flow cytometry within 1 hour.

Determination of the content of Collagen I and Collagen III: collect the cell supernatants after 24 hours of treatment with each concentration of the drug group (including the control group), conduct experiments according to the instructions of the ELISA kit, and finally measure the absorbance value at a wavelength of 540 nm on a microplate reader. Draw a standard curve and calculate the content of Collagen I and Collagen III in each group.

Immunofluorescence method to detect the expression of α-SMA: after the cell slides were fixed, fluorescent antibodies were added dropwise, stained at 37°C for 30 minutes (keep moist), washed twice with PBS, washed once with distilled water, sealed with 50% glycerol, examined under a microscope and photographed .

: After the cells were treated with drugs of various concentrations for 24 hours, wash them with PBS for 3 times, add 1mL Trizol to each well after drying, let them rest for 10 minutes, then blow them properly, and collect and extract RNA after all the cells are broken. Measure the A260/A280 value with a UV spectrophotometer, adjust to a uniform concentration, use a reverse transcription kit and reverse transcription instrument to reverse transcribe it into cDNA and dilute it, perform PCR, and perform agarose gel electrophoresis on the product to observe the specificity of each product and expression. The expression levels of EGFR, EGF, VEGF, Collagen I, Collagen III, MMP, TIMP-1, α-SMA and other mRNAs were calculated by relative quantitative method.

After the cells were treated with drugs of various concentrations for 24 hours, they were washed with PBS for 3 times, and the total protein was extracted by adding RIPA lysate to the control and pulverized. The primary antibody was added behind the membrane and reacted overnight at 4°C. After washing the membrane the next day, the secondary antibody was added and incubated at room temperature for 1 h. ECL self-image on X-ray film, observe and analyze the gray density value of each electrophoresis band after washing the film. The expressions of EGFR, EGF, VEGF, Collagen I, Collagen III, MMP, TIMP-1, α-SMA, Smad, PI3K, AKT and ERK were calculated by relative quantitative method.

test results:

1. The MTT experimental data of some optimized matrine derivatives on various cell lines are shown in the figure,

Figure 2 shows the effect of matrine derivatives on the proliferation of liver cancer cell line Hep-G2. Compared with the negative control group, the drugs with P<0.05 are: WM-101, WM-202, WM-204. The drugs with P<0.01 are: M-19, WM-102, WM-103, WM-104, WM-105, WM-106, WM-107, WM-108, WM-110, WM-111, WM-112, WM-113, WM- 114, WM-115, WM-116, WM-117, WM-119, WM-121, WM-122, WM-123, WM-124, WM-125, WM-127, WM-128, WM-129, WM-130, WM-132, WM-133, WM-134, WM-201, WM-203, WM-301, WM-302, WM-303, WM-304, WM-313, RM-1

Figure 3 is the effect of matrine derivatives on the proliferation of liver cancer cell line Hun-7, compared with the negative control group

The drugs with P<0.05 are: WM-107, WM-126

The drugs with P<0.01 are: WM-127, WM-128, WM-129, WM-130, WM-133, RM-1

Figure 4 is the effect of matrine derivatives on the proliferation of liver cancer cell line MHCC-97L

Compared with the negative control group

Drugs with P<0.05: WM-104, WM-106, WM-107, WM-112

The drugs with P<0.01 are: M-19, WM-123, WM-127, WM-129, WM-130, WM-132

Figure 5 is the effect of matrine derivatives on the proliferation of liver cancer cell line SMMC-7721

Compared with the negative control group

Drugs with p<0.05: WM-133

The drugs with p<0.01 are: WM-107, WM-121, WM-127, WM-129, WM-130.

Figure 6 is the effect of matrine derivatives on the proliferation of liver cancer cell line Hep-3B

Compared with the negative control group

Drugs with p<0.05: WM-203

The drugs with p<0.01 are: WM-102, WM-107, WM-122, WM-123, WM-129, WM-130, WM-133, WM-201, WM-202, WM-302, M-19 .

Figure 7 is the effect of matrine derivatives on fibroblast BJ proliferation

Compared with the negative control group

The drugs with P<0.05 are: WM-106, WM-115, WM-116, WM-117, WM-132, WM-134, WM-201, WM-202, WM-301

The drugs with P<0.01 are: M-19, WM-101, WM-105, WM-107, WM-112, WM-113, WM-114, WM-119, WM-121, WM-122, WM-126 , WM-127, WM-128, WM-129, WM-130, WM-133, WM-314, ACM-1

Figure 8 is the effect of matrine derivatives on the proliferation of human hepatic stellate cells LX-2

Compared with the negative control group

The drugs with P<0.05 are: WM-105, WM-110, WM-115, WM-116, WM-123, WM-128, WM-134

The drugs with P<0.01 are: M-19, WM-101, WM-104, WM-106, WM-107, WM-111, WM-112, WM-119, WM-125, WM-126, WM-127 , WM-129, WM-130, WM-132, WM-133, WM-202, WM-204, WM-301, WM-302, WM-303, RM-1

Figure 9 is the effect of matrine derivatives on the proliferation of rat hepatic stellate cells HSC-T6

Compared with the negative control group

Drugs with P<0.05: WM-111, WM-125, WM-313, WM-128

The drugs with P<0.01 are: M-19, WM-101, WM-102, WM-103, WM-104, WM-105, WM-106, WM-107, WM-108, WM-110, WM-112 , WM-113, WM-114, WM-115, WM-116, WM-117, WM-119, WM-122, WM-123, WM-126, WM-127, WM-129, WM-130, WM -132, WM-133, WM-134, WM-201, WM-202, WM-203, WM-204, WM-301, WM-302, WM-303, WM-304, WM-314, ACM-1

2. Effect of matrine derivative WM-108 on Hep3B cells

(1) Matrine derivative WM-108 inhibits the proliferation of Hep3B (see Table 5 and Figure 10)

Table 5. The effect of WM-108 on inhibiting the proliferation of Hep3B

Figure 10 shows the effect of WM-108 on inhibiting Hep3B. As the concentration increases, the inhibition rate of WM-108 on Hep3B also gradually increases

(2) Matrine derivative WM-108 inhibits Hep3B migration and invasion (see Figure 11)

Figure 11 shows that WM-108 inhibits Hep3B migration and invasion. As the concentration increases, cell migration and invasion are significantly inhibited

3. The effect of matrine derivative WM130 on rat hepatic stellate cells (HSC-T6) and human hepatic stellate cells (LX-2)

(1) WM130 inhibits the proliferation of HSC-T6 and LX-2

Table 6. The effect of WM130 on inhibiting the proliferation of HSC-T6 and LX-2

Note: ^* p < 0.05, ^** p < 0.01.

(2) The effect of WM130 on the proliferation and morphology of LX-2 cells (see Figure 12)

Figure 12 shows the cell morphology of LX-2 cells after different concentrations of WM130 acted on them for 24 hours. As the concentration increased, the cell proliferation slowed down, and the cell shape became smaller and rounder.

The above is only a preferred embodiment of the present invention, it should be pointed out that for those of ordinary skill in the art, without departing from the method of the present invention, some improvements and supplements can also be made, and these improvements and supplements should also be considered Be the protection scope of the present invention.

Claims (7)

1. class matrine derivative and a salt thereof, its structure is as shown in general formula:

Wherein X is sulphur atom;

Wherein n is selected from 1 or 2;

Wherein R ₁, R ₂be selected from or or :

. alkyl, is selected from methoxyl group, oxyethyl group, positive propoxy, isopropoxy, n-butoxy, isobutoxy or benzyloxy, preferably methoxyl group;

. aryl, be selected from benzyl, to luorobenzyl, p-chlorobenzyl, adjacent luorobenzyl, to methyl-benzyl, preferably to methyl-benzyl;

. ring is amino, and ring amino is piperidyl, Pyrrolidine base and piperazinyl.

2. matrine derivative according to claim 1 and salt thereof, is characterized in that: described X is sulphur atom.

3. matrine derivative according to claim 1 and salt thereof, is characterized in that: described X is sulphur atom, and n is 1 or 2.

4. matrine derivative according to claim 1 and salt thereof, is characterized in that: described matrine derivative is 13-[ n-methyl, n-(2-allylamino ethanoyl)]-amino-15-sulfo-matrine.

5. matrine derivative according to claim 1 and salt thereof, is characterized in that: described matrine derivative is 13-[ n-methyl, n-(4 fluorophenethyl glycyl)]-amino-15-sulfo-matrine.

6. matrine derivative according to claim 1 and salt thereof, is characterized in that: described salt is hydrochloride, vitriol, hydrosulfate, hydrobromate, oxalate, Citrate trianion or mesylate.

7. the matrine derivative as described in as arbitrary in claim 1-6 and the preparation method of salt thereof, the method is as follows;

the synthetic route of WM-1 series compound:

the synthetic route of WM-2 series compound:

the synthetic route of WM-3 series compound:

The synthetic of the compounds of this invention salt is on the basis of above-mentioned reaction, further does following reaction:

HA is hydrochloric acid, sulfuric acid, hydrogen sulfate, Hydrogen bromide, oxalic acid, citric acid or methylsulfonic acid.