Berberine salt and application thereof

Abstract

The invention discloses berberine salt, belongs to the technical field of medical preparations, and particularly relates to berberine dialkyl acid salt (also called dialkyl acid salt berberine) which specifically comprises dibutyrate berberine and diacetate berberine. Compared with monoalkyl acid salt berberine, the dialkyl acid salt berberine has different chemical properties and physical properties; the quality is controllable, no difference exists between batches, and the stability is good; the dialkyl acid salt berberine has a better effect on improving the metabolic function and inhibiting tumor cell proliferation, and the pharmacological action is enhanced.

Description

Technical Field

[0001] The present invention relates to the technical field of medical preparations, and particularly relates to a berberine salt and its application. Background Art

[0002] Berberine (molecular formula: C 20 H 18 NO4, molecular weight: 336.37) is a quaternary ammonium alkaloid isolated from the traditional Chinese medicine Coptis chinensis, and is the main active ingredient for the antibacterial effect of Coptis chinensis.

[0003] Its clinical application is mainly as an over-the-counter drug for preventing and treating dysentery. Berberine has a broad antibacterial spectrum and has antibacterial effects on a variety of Gram-positive and negative bacteria in vitro. For example, it has inhibitory effects on Shigella dysenteriae, Mycobacterium tuberculosis, Streptococcus pneumoniae, Salmonella typhi, and Corynebacterium diphtheriae, etc. It has the strongest effect on Shigella dysenteriae, and there is almost no drug resistance and side effects. In recent years, with the continuous in-depth research, its scope of application has had new developments. Modern pharmacological research has confirmed that berberine also has significant effects in regulating blood sugar and lipid metabolism, anti-inflammatory, reducing blood uric acid, anti-rheumatoid arthritis, and inhibiting the proliferation of tumor cells, antiviral, etc. A number of basic and clinical evidences have confirmed that berberine participates in glucose metabolism through multiple mechanisms.

[0004] Berberine is a yellow needle-shaped crystal with an extremely bitter taste. Free berberine can slowly dissolve in hot water or hot ethanol, but has a small solubility in cold water and is almost insoluble in cold ethanol, and is almost insoluble in chloroform and ether. The hydrochloride salt of berberine is easily soluble in boiling water, slightly soluble in cold water, and hardly soluble in ethanol, and its water solubility cannot meet the requirements. The solubility of berberine has an important impact on its application in drugs. Therefore, developing berberine salts and enhancing the pharmacological effects and curative effects of berberine have become important research directions.

[0005] The preparation of mono-alkylates of berberine has been reported in the literature. However, a set of impurity peaks can be clearly seen in the NMR spectrum of the mono-alkylates reported in the literature. In subsequent studies, it was found that this impurity inevitably exists, and the content of this impurity is inconsistent among different batches of mono-alkylates of berberine, resulting in uncontrollable quality of berberine mono-alkylates and unable to carry out further development research. Summary of the Invention

[0006] Aiming at the above technical problems existing in the prior art, the present invention provides a berberine salt and its application, which enhances the pharmacological effects of berberine and is stable and controllable in quality.

[0007] The present invention discloses a berberine salt, including berberine dialkylate.

[0008] Preferably, the structure of berberine dialkylate is represented as: ; Among them, R represents an alkyl group, and n can take values of 0.5, 1, 1.5, or 2, but is not limited thereto.

[0009] Preferably, the berberine dialkylates are selected from: berberine dibutyrate and berberine diacetate; The structure of berberine dibutyrate is represented as: .

[0010] Preferably, the preparation method of the berberine dialkylates includes: Dissolve berberine acetone in ethanol, add an alkyl acid under stirring for reaction, filter after the reaction ends, concentrate and dry the filtrate to obtain a concentrated solution; Add ethyl acetate to the concentrated solution, filter and dry to obtain berberine alkylate; Take the berberine alkylate, add an aqueous solution of the alkyl acid, shake, then rotary evaporate and dry to obtain the berberine dialkylate.

[0011] Preferably, the preparation method of berberine acetone includes: Add methanol to berberine hydrochloride, and stir to obtain a suspension; Add a sodium hydroxide solution to the suspension, and stir to obtain a second solution; After dropping acetone into the second solution, drop water, stir and then filter to obtain a filter cake; Wash the filter cake and then dry to obtain berberine acetone.

[0012] Preferably, the preparation method of berberine dibutyrate includes: At room temperature, suspend 409 mg of berberine hydrochloride in 10 mL of methanol to obtain a suspension; Add 2 mL of 0.5 N sodium hydroxide solution to the suspension, stir at the same temperature for 30 min to obtain a reddish-brown second solution; After dropping 2 ml of acetone into the second solution, slowly drop 4 ml of water, yellow precipitates will precipitate out. Stir at room temperature for 1 h, then filter to obtain a filter cake; Wash the filter cake with a 1:1 methanol aqueous solution and then dry to obtain berberine acetone; Dissolve 196.5 mg of berberine acetone in 0.25 ml of absolute ethanol, heat to 45 °C, add 0.5 mmol of butyric acid under stirring, stop heating after reacting for 10 hours, filter while it is hot, and concentrate and dry the filtrate; After drying, add ethyl acetate to precipitate a brownish-yellow solid, filter and then dry to obtain the crude product of berberine butyrate; Add 0.5 mL of 10% butyric acid aqueous solution to 20 mg of the crude product of berberine butyrate, shake for 30 min, then concentrate and dry at 60 °C to obtain berberine dibutyrate.

[0013] Preferably, the preparation method of berberine diacetate includes: At room temperature, 409 mg of berberine hydrochloride is suspended in 10 mL of methanol to obtain a suspension; 2 mL of 0.5 N sodium hydroxide solution is added to the suspension, and the mixture is stirred at the same temperature for 30 min to obtain a second reddish-brown solution; After adding 2 ml of acetone to the second solution, 4 ml of water is slowly added dropwise, and yellow precipitates are separated out. After stirring at room temperature for 1 h, filtration is carried out to obtain a filter cake; After washing the filter cake with a 1:1 methanol-water solution, it is dried to obtain berberine acetone; 196.5 mg of berberine acetone is dissolved in 0.25 ml of absolute ethanol, heated to 45 °C, and 0.5 mmol of acetic acid is added under stirring. After reacting for 10 hours, heating is stopped, and filtration is carried out while it is hot. The filtrate is concentrated and dried; After drying, ethyl acetate is added to precipitate a yellowish-brown solid, and after filtration, it is dried to obtain a crude product of berberine acetate; 0.5 mL of 10% acetic acid aqueous solution is added to 20 mg of the crude product of berberine acetate, shaken for 30 min, and then concentrated and dried at 60 °C to obtain berberine diacetate.

[0014] Preferably, berberine dialkylate is used to improve metabolism and inhibit the proliferation of tumor cells.

[0015] Preferably, berberine dialkylate is used to reduce body weight, lower serum total cholesterol, triglyceride, low-density lipoprotein-cholesterol, and lower blood glucose.

[0016] The present invention also provides the application of the above-mentioned berberine salts. Berberine dialkylate is used to prepare a drug for improving metabolic function or a drug for anti-tumor cell proliferation.

[0017] Compared with the prior art, the beneficial effects of the present invention are as follows: Compared with berberine monoalkylate, berberine dialkylate has different chemical properties and physical properties; it is stable, the quality is controllable, and there is no difference between batches; berberine dialkylate has better effects in improving metabolic function and inhibiting the proliferation of tumor cells, enhancing the pharmacological effect. Description of the Drawings

[0018] Figure 1 is the hydrogen spectrum detection chart of berberine dibutyrate; Figure 2 is the carbon spectrum detection chart of berberine dibutyrate; Figure 3 is the DSC chart of berberine dibutyrate; Figure 4 is the hydrogen spectrum detection chart of berberine diacetate; Figure 5 It is the powder X-ray diffraction pattern of berberine dibutyrate; Figure 6 It is the hydrogen spectrum of berberine monobutyrate in the comparative example; Figure 7 It is the carbon spectrum of berberine monobutyrate in the comparative example; Figure 8 It is the DSC diagram of berberine monobutyrate; Figure 9 It is the powder X-ray diffraction pattern of berberine monobutyrate; Figure 10 It is the weight change curve graph of the animal experiment; Figure 11 It is the bar graph of the weight at the 8th week; Figure 12 It is the comparison graph of the epididymal fat weight of mice; Figure 13 It is the bar graph of the plasma cholesterol content; Figure 14 It is the bar graph of the plasma low density lipoprotein-cholesterol content; Figure 15 It is the bar graph of the plasma triglyceride TG content; Figure 16 It is the bar graph of the blood glucose content; Figure 17 It is the bar graph of the alanine aminotransferase level; Figure 18 It is the bar graph of the aspartate aminotransferase level; Figure 19 It is the bar graph of the liver triglyceride T content; Figure 20 It is the proliferation curve graph of the tumor cells HCT-116 in each group; Figure 21 It is the proliferation curve graph of the tumor cells HT29 in each group; Figure 22 It is the flow chart of the preparation method of the berberine salt of the present invention. Specific embodiments

[0019] To make the objectives, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are some, but not all, of the embodiments of the present invention. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the protection scope of the present invention.

[0020] The present invention will be further described in detail below in conjunction with the accompanying drawings: The present invention provides a berberine salt, berberine dialkylate (also known as dialkylate berberine). Specifically, it includes berberine dibutyrate (also known as dibutyrate berberine) or berberine diacetate.

[0021] The structure of berberine dialkylate is represented as: ; wherein, R represents an alkyl group, and n can take values of 0.5, 1, 1.5 or 2, but is not limited thereto.

[0022] The structural formula of berberine dibutyrate is represented as: ; wherein, berberine combines with two butyrate groups.

[0023] As Figure 22 shown, the preparation method of berberine dibutyrate includes the following steps: Step 101: Add methanol to berberine hydrochloride, and stir to obtain a suspension.

[0024] Step 102: Add a sodium hydroxide solution to the suspension, and stir to obtain a second solution, which is a clear solution.

[0025] Step 103: Dropwise add acetone to the second solution, then add water dropwise, stir and filter to obtain a filter cake.

[0026] Step 104: Wash the filter cake and then dry it to obtain acetone berberine 2.

[0027] Step 105: Dissolve acetone berberine in ethanol, add an alkyl acid to react under stirring, filter after the reaction ends, concentrate and dry the filtrate to obtain a concentrated solution.

[0028] In specific preparation, the alkyl acid used is butyric acid or acetic acid.

[0029] Step 106: Add ethyl acetate to the concentrated solution, filter and dry to obtain berberine alkylate 3.

[0030] Step 107: Take berberine alkylate, add an aqueous solution of an alkyl acid, shake, then rotary evaporate and dry to obtain berberine dialkylate 4.

[0031] Correspondingly, in specific preparation, berberine dibutyrate and berberine diacetate are respectively obtained.

[0032] Reaction formula 1 roughly reflects the above reaction process: (1); wherein, R represents an alkyl group, preferably a C2-C6 alkyl group.

[0033] Example 1: Preparation of berberine dibutyrate, including the following steps: Step 201: At room temperature, suspend 409 mg (1.1 mmol) of berberine hydrochloride in 10 mL of methanol to obtain a suspension.

[0034] Step 202: Add 2 mL of 0.5 N sodium hydroxide solution to the suspension, stir at the same temperature for 30 min to obtain a reddish-brown solution, i.e., the second solution.

[0035] Step 203: After adding 2 ml of acetone to the second solution, slowly add 4 ml of water, yellow precipitate will precipitate out. After stirring at room temperature for 1 h, filter to obtain the filter cake.

[0036] Step 204: Wash the filter cake with a small amount of methanol / water (1:1), dry to obtain yellow solid berberine acetone 2 (370 mg, yield 88.9%).

[0037] Step 205: Weigh 196.5 mg of berberine acetone 2 (0.5 mmol), dissolve it in 0.25 ml of anhydrous ethanol, heat to 45 °C, add 0.5 mmol of butyric acid under stirring, stop heating after reacting for 10 h, filter while it is hot, and concentrate and dry the filtrate.

[0038] Step 206: After drying, add ethyl acetate to precipitate a brownish-yellow solid, filter and dry to obtain the crude product of berberine butyrate (177.5 mg, yield 83.9%).

[0039] Step 207: Add 0.5 mL of 10% aqueous butyric acid solution to 20 mg of the crude product of berberine butyrate, shake for 30 min, concentrate and dry at 60 °C to obtain berberine dibutyrate (22 mg, yield 91.7%).

[0040] Detection of berberine dibutyrate: Figure 1 The hydrogen spectrum detection results of berberine dibutyrate are shown, Figure 2 The carbon spectrum detection results of berberine dibutyrate are shown.

[0041] The nuclear magnetic resonance detection results are as follows: 1 H NMR (500 MHz, DMSO-d6) δ 9.91 (s, 1H), 8.94(s, 1H), 8.20 (d, J = 9.1 Hz, 1H), 8.00 (d, J = 9.0 Hz, 1H), 7.79 (s, 1H),7.08 (s, 1H), 6.17 (s, 2H), 4.94 (t, J= 6.3 Hz, 2H), 4.10 (s, 3H), 4.07 (s,3H), 3.21 (t, J = 6.2 Hz, 2H), 1.99 (t, J = 7.4 Hz, 4H), 1.50 - 1.38 (m, 4H),0.82 (t, J = 7.3 Hz, 6H); 13 C NMR (500 MHz, DMSO-d6) δ 175.08, 150.30, 149.74,147.61, 145.40, 143.60, 137.41, 132.91, 130.60, 126.69, 123.43, 121.32,120.36, 120.11, 108.33, 105.35, 101.99, 61.82, 56.97, 55.09, 38.06, 26.25,18.71, 13.92. Element analysis:calculated element composition (C, 65.74%; H,6.50%; N, 2.74%; O, 25.02%), experimental determined values (C, 64.97%; H,6.15%; N, 2.79%; O, 25.09%).

[0042] Figure 3 showed the DSC (Differential scanning calorimetry) spectrum. Berberine dibutyrate showed an obvious endothermic peak only at 135.1 o °C, which was consistent with dibutyric acid. Figure 5 showed the powder X-ray diffraction results of berberine dibutyrate.

[0043] In multiple repeated preparations, there were no differences between batches, it was stable and the quality was controllable, and further development and research could be carried out.

[0044] Example 2: Preparation of berberine acetate. The preparation method refers to steps 201 - 207 of Example 1, replacing butyric acid with acetic acid to obtain berberine diacetate.

[0045] The nuclear magnetic resonance detection results were as follows: 1 H NMR (500 MHz, DMSO-d6) δ9.91 (s, 1H), 8.95(s, 1H), 8.21 (d, J J = 9.1 Hz, 1H), 8.00 (d, J J = 9.1 Hz, 1H), 7.80 (s, 1H),7.09 (s, 1H), 6.18 (s, 2H), 4.94 (t, J J = 6.4 Hz, 2H), 4.10 (s, 3H), 4.07 (s,3H), 3.21 (t, J J = 6.2 Hz, 2H), 1.67 (s, 5H). Figure 4 The hydrogen spectrum detection results of berberine diacetate are shown.

[0046] Comparative example: The method for preparing berberine monobutyrate includes: dissolving a mixture of berberine hydrochloride (100.0 mg, 269.0 mmol) and sodium butyrate (44.4 mg, 403.0 mmol) in water (20 ml). After stirring at 80 °C for 60 min, it was cooled for crystallization, placed in a refrigerator at -4 °C for more than 12 hours, and the solid was filtered off. The filtrate was concentrated into an amorphous solid. After washing the solid with anhydrous ethanol (20 ml × 3 times), it was dried in vacuo to obtain berberine monobutyrate, also known as butyric acid berberine or berberine monobutyrate.

[0047] Berberine monobutyrate was detected. From Figure 6 and Figure 7 the 1H NMR and 13C NMR spectra of Figure 8 it can be seen that the purity of this compound is not high and there is an obvious set of impurity peaks; in addition, from Figure 3 the DSC spectrum of 1 H NMR (500 MHz, DMSO-d6) δ 9.91 (s, 1H), 8.95 (s, 1H), 8.20 (d, J J = 9.1 Hz, 1H), 8.01 (d, J J = 9.0 Hz, 1H), 7.81 (s, 1H), 7.09 (s, 1H), 6.17(s, 2H), 4.94 (t, J J = 6.3 Hz, 2H), 4.10 (s, 3H), 4.07 (s, 3H), 3.21 (t, J= 6.3 Hz, 2H), 1.83 (t, J = 7.3 Hz, 2H), 1.44 - 1.34 (m, 2H), 0.79 (t, J = 7.4 Hz, 3H); 13 C NMR (500 MHz, DMSO-d6) δ 174.95, 150.28, 149.70, 147.57, 145.43, 143.57, 137.36, 132.89, 130.56, 126.59, 123.47, 121.30, 120.36, 120.13, 108.30, 105.37, 101.97, 61.81, 56.94, 55.04, 39.01, 26.25, 19.33, 14.26. Elemental analysis results: N 3.01%, C 60.25%, H 6.108%, O% 28.593. Figure 9 The powder X-ray diffraction results of berberine butyrate are shown.

[0048] Metabolic function regulation detection. The berberine salts of the present invention can be used to prepare drugs for treating metabolic-related diseases such as obesity, hyperglycemia, and hyperlipidemia. By feeding C57 mice a high-fat diet feed, a metabolic disease model is established.

[0049] Detection of the effect on body weight. C57 mice (5 weeks old) were randomly divided into 6 groups, namely a control group (normal diet, NCD), a model group (high-fat diet, HFD), and 4 drug administration groups: berberine hydrochloride (BBR) was given simultaneously with a high-fat diet, a physical mixture of berberine hydrochloride and sodium butyrate (B2NM, 1:1), berberine monobutyrate (B1BS), and berberine dibutyrate (B2BS); there were 8 mice in each group, and the grouping situation is shown in Table 1. The animals in each group were given intragastric administration. The mice in the control group and the model group were given 10 mL / kg / d distilled water. The drug administration doses of the other groups are shown in Table 1. The drug was administered once a day for 8 consecutive weeks. After the drug administration was completed, the animals were anesthetized and sacrificed by intraperitoneal injection of 1 ml of 20% chloral hydrate. Blood was collected by eye socket puncture, and the liver and epididymal fat were taken. An automatic biochemical analyzer was used to measure blood biochemical indexes. The experimental results show that B2BS has the effects of reducing blood sugar, reducing blood lipids, and reducing triglycerides in the liver, and the effects are better than those of BBR, B2NM, and B1BS. Therefore, B2BS can effectively prevent and treat hyperglycemia, hyperlipidemia, and metabolic fatty liver disease.

[0050] Table 1

[0051] Effect of Berberine Dibutyrate on the Body Weight of Experimental Animals. The body weight of mice was recorded weekly during the experiment, and its weight change is shown as Figure 10 follows. Figure 11 Figure Figure 11 shows the weight comparison at the 8th week of drug intervention. As can be seen from the above figure, compared with normal diet, high-fat diet significantly increased the body weight of C57 mice. Compared with the model group, Berberine Dibutyrate of B2BS could significantly reduce the body weight gain caused by high-fat diet, and the degree of weight reduction was better than that of BBR, B2NM and B1BS.

[0052] Detection of the effect on epididymal fat. The detection results of the epididymal fat weight of each group are shown in Table 2 and Figure 12 .

[0053] Table 2

[0054] After high-fat diet modeling, the ratio of epididymal fat / body weight of animals increased significantly. Each drug administration group had the effect of reducing the epididymal fat weight, and the degree of reducing epididymal fat by B2BS was better than that of BBR, B2NM and B1BS.

[0055] Effect on blood lipid and blood glucose of experimental animals. The detection of blood lipid mainly targeted the contents of cholesterol, low-density lipoprotein and triglyceride in plasma. Table 3 shows the detection results of plasma cholesterol (CHO) content; Figure 13 Figure Figure 13 shows the bar chart of cholesterol content.

[0056] Table 3

[0057] The content of plasma low-density lipoprotein-cholesterol (LDL-c) is shown in Table 4 and Figure 14 .

[0058] Table 4

[0059] The results of plasma triglyceride TG content are shown in Table 5 and Figure 15 .

[0060] Table 5

[0061] The detection results of blood glucose content are shown in Table 6 and Figure 16 .

[0062] Table 6

[0063] After high-fat diet-induced modeling, blood glucose, cholesterol, low-density lipoprotein, and triglyceride levels in the plasma of animals increased significantly. Each drug-administered group had the effect of reducing blood lipid and blood glucose levels. B2BS could effectively reduce the increase in blood glucose and blood lipid caused by high-fat diet, and the degree of reduction was better than that of BBR, B2NM, and B1BS.

[0064] Detection of the effects on the liver function of experimental animals. Table 7 and Figure 17 show the levels of alanine aminotransferase (ALT) in mice of each group after intervention. It can be Figure 17 seen that compared with the negative control group, the ALT level in the model group increased significantly. Compared with the model group, the B2BS group significantly reduced the ALT level caused by high-fat diet, and the degree of reduction was better than that of BBR, B2NM, and B1BS.

[0065] Table 7

[0066] Table 8 and Figure 18 show the detected levels of aspartate aminotransferase (AST) in mice of each group after the end of the intervention. Compared with the negative control group, the AST level in the model group increased significantly. Compared with the model group, the B2BS group significantly reduced the AST level caused by high-fat diet, and the degree of reduction was better than that of BBR, B2NM, and B1BS.

[0067] Table 8

[0068] Table 9 and Figure 19 show the measurement results of the content of triglyceride (TG) in the liver of mice of each group after the end of the intervention.

[0069] Table 9

[0070] Detection of anti-tumor effects. The stock solution doses were prepared as follows: berberine hydrochloride (BBR, 10 mM, with water as the solvent), dibutyrate berberine (B2BS, 20 mM, with DMSO as the solvent), monobutyrate berberine (B1BS, 20 mM, with DMSO as the solvent), and sodium butyrate (NaB, 40 mM, with water as the solvent).

[0071] Two different human intestinal cancer cell lines (HCT-116, HT-29) were used for in vitro experiments, and the CCK8 method was used to detect the cell growth curve.

[0072] The final dosing doses and groups were as follows: negative control group with an equal amount of culture medium, the first control group (20 μM berberine hydrochloride, BBR), the second test group (20 μM dibutyrate berberine group, B2BS), the first test group (20 μM monobutyrate berberine, B1BS), and the second control group (physical mixture of 20 μM berberine hydrochloride and 20 μM sodium butyrate, B2NM).

[0073] Effects on the proliferation of tumor cells HCT-116 and HT-29 Figure 20 and Figure 21 In [the experiment], the absorbance OD value reflects the cell density in the culture medium, and specifically OD600 was used. Figure 20 In the proliferation detection of HCT-116 cells, dibutyrate berberine significantly inhibited the proliferation of tumor cells, and its effect was better than that of berberine hydrochloride, the mixture of berberine hydrochloride and sodium butyrate, and monobutyrate berberine at the same molar ratio. In Figure 21 the proliferation detection of HT29 cells, dibutyrate berberine significantly inhibited the proliferation of tumor cells, and its effect was better than that of berberine hydrochloride, the mixture of berberine hydrochloride and sodium butyrate, and monobutyrate berberine at the same molar ratio.

[0074] In summary, compared with monobutyrate berberine, the dibutyrate berberine of the present invention has changed physical and chemical properties, controllable quality, no difference between batches, and good stability; it has better effects in improving metabolic function and inhibiting the proliferation of tumor cells. Specifically, in animal experiments, it showed excellent effects in reducing body weight, lowering serum total cholesterol (TC), triglyceride (TG), low-density lipoprotein-cholesterol (LDL-c), and blood glucose. In cell experiments, it showed excellent effects in inhibiting tumor proliferation. Therefore, the dialkyl berberine salts of the present invention can be used to prepare drugs for improving metabolic function or drugs for inhibiting the proliferation of anti-tumor cells.

[0075] The dosage form of the pharmaceutical composition can be a liquid dosage form, a solid dosage form, or a semi-solid dosage form. By combining berberine salts with one or more pharmaceutically acceptable solid or liquid excipients, any dosage form suitable for human or animal use can be prepared. The berberine salts or pharmaceutical compositions containing them can be administered in unit dosage form, and the administration routes can be enteral or parenteral, such as oral, intravenous injection, intramuscular injection, subcutaneous injection, nasal cavity, oral mucosa, eye, lung and respiratory tract, skin, vagina, rectum, etc. In addition, if necessary, colorants, preservatives, fragrances, flavoring agents, or other additives can also be added to the pharmaceutical preparation.

[0076] The cardiovascular pharmacological effects of the berberine salts of the present invention mainly include anti-arrhythmia, anti-heart failure, vasodilation and blood pressure reduction, regulation of lipid metabolism and anti-atherosclerosis, anti-platelet and anti-thrombosis, anti-cerebral ischemia, protection of vascular endothelium, anti-vascular inflammation and inhibition of tumor neovascularization. The target cells of the action include cardiomyocytes, vascular endothelial cells, vascular smooth muscle cells, platelets and blood mononuclear macrophages, etc. The specific targets are mainly Ca2+ channels, K+ channels, M receptors, adenosine receptors, cholinesterase and ACE, etc.

[0077] A number of basic and clinical evidences confirm that berberine salts are involved in glucose metabolism through multiple mechanisms. Such as the retinol binding protein-4 (RBP-4) and glucose transporter-4 (GLUT-4) mechanisms; increasing the expression of hepatocyte nuclear factor-4a (HNF-4a) and glucose kinase activity; reversing the phosphorylation of insulin receptor (IRS)-1 Ser307; up-regulating the expression of IRS, etc. The lipid-lowering function of berberine salts, the molecular mechanism involves up-regulating the level of LDLR-mRNA, inhibiting the expression of 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR) gene, increasing the expression of hepatic apolipoprotein (Apo) E mRNA, activating the AMPK pathway, sterol regulatory element binding protein (SREBP)-C and CCAAT / enhancer binding protein (C / EBP)-α, peroxisome proliferator-activated receptor (PPAR)-γ, reducing the transcriptional level of proprotein convertase subtilisin / kexin type 9 (PCSK) 9 gene, etc.

[0078] The above are only the preferred embodiments of the present invention and are not intended to limit the present invention. For those skilled in the art, the present invention can have various changes and modifications. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A berberine salt, characterized in that, It includes berberine dialkylates.

2. The berberine salt according to claim 1, wherein The structure of berberine dialkylates is represented as: ； wherein, R represents an alkyl group.

3. The berberine salt according to claim 1, wherein The berberine dialkylates are selected from: berberine dibutyrate and berberine diacetate; wherein, the structure of berberine dibutyrate is represented as: 。 4. The berberine salt according to claim 3, characterized in that, The preparation method of berberine dialkylates includes: Dissolve berberine acetone in ethanol, add alkyl acid under stirring for reaction, filter after the reaction ends, concentrate and dry the filtrate to obtain a concentrated solution; Add ethyl acetate to the concentrated solution, filter and dry to obtain berberine alkylate; Add an aqueous solution of alkyl acid to the berberine alkylate, shake, then rotary evaporate and dry to obtain berberine dialkylates.

5. The berberine salt according to claim 4, characterized in that, The preparation method of berberine acetone includes: Add methanol to berberine hydrochloride, stir to obtain a suspension; Add sodium hydroxide solution to the suspension, stir to obtain a second solution; Dropwise add acetone to the second solution, then dropwise add water, stir and filter to obtain a filter cake; Wash the filter cake and then dry to obtain berberine acetone.

6. The berberine salt according to claim 3, wherein The preparation method of berberine dibutyrate includes: At room temperature, suspend 409 mg of berberine hydrochloride in 10 mL of methanol to obtain a suspension; Add 2 mL of 0.5 N sodium hydroxide solution to the suspension, stir at the same temperature for 30 minutes to obtain a second solution; Dropwise add 2 ml of acetone to the second solution, slowly dropwise add 4 ml of water, stir at room temperature for 1 hour, then filter to obtain a filter cake; Wash the filter cake with a 1:1 methanol aqueous solution and then dry to obtain berberine acetone; Dissolve 196.5 mg of berberine acetone in 0.25 ml of anhydrous ethanol, heat to 45 °C, add 0.5 mmol of butyric acid under stirring, stop heating after reacting for 10 hours, filter while it is hot, concentrate and dry the filtrate; After drying, add ethyl acetate, filter and dry to obtain the crude product of berberine butyrate; Add 0.5 mL of 10% butyric acid aqueous solution to 20 mg of the crude product of berberine butyrate, shake for 30 minutes, then concentrate and dry at 60 °C to obtain berberine dibutyrate.

7. The berberine salt according to claim 3, characterized in that, The preparation method of berberine diacetate includes: At room temperature, suspend 409 mg of berberine hydrochloride in 10 mL of methanol to obtain a suspension; Add 2 mL of 0.5 N sodium hydroxide solution to the suspension, stir at the same temperature for 30 minutes to obtain a second solution; Dropwise add 2 ml of acetone to the second solution, slowly dropwise add 4 ml of water, stir at room temperature for 1 hour, then filter to obtain a filter cake; Wash the filter cake with a 1:1 methanol aqueous solution and then dry to obtain berberine acetone; Dissolve 196.5 mg of berberine acetone in 0.25 ml of anhydrous ethanol, heat to 45 °C, add 0.5 mmol of acetic acid under stirring, stop heating after reacting for 10 hours, filter while it is hot, concentrate and dry the filtrate; After drying, add ethyl acetate, filter and dry to obtain the crude product of berberine acetate; Add 0.5 mL of 10% acetic acid aqueous solution to 20 mg of the crude product of berberine acetate, shake for 30 minutes, then concentrate and dry at 60 °C to obtain berberine diacetate.

8. The berberine salt according to claim 1, wherein Berberine dialkylates are used to improve metabolism and inhibit the proliferation of tumor cells.

9. The berberine salt according to claim 8, wherein, Dialkyl berberine salts are used for weight loss, reducing total serum cholesterol, triglycerides, low-density lipoprotein-cholesterol, and blood glucose.

10. Use of the berberine salt according to any one of claims 1-9, characterized in that, Dialkyl berberine salts are used for preparing drugs for improving metabolic function or drugs for anti-tumor cell proliferation.

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