A centella asiatica glycoside, leonurine and polygonatum polysaccharide pharmaceutical composition and application thereof

Abstract

The application discloses a centella asiatica glycoside, motherwort alkaloid and polygonatum sibiricum polysaccharide medicine composition and application, and can treat diabetes and diabetic foot and other difficult wound healing caused by diabetes. Pharmacodynamic experiments show that the composition can effectively reduce blood sugar, has clear curative effect and positive significance on improvement of symptoms such as tail amputation regeneration and difficult wound healing of simulated diabetic foot, and the prescription proportion of the composition is optimized. The medicine composition has the advantages of simple prescription, high active ingredient content, safety and effectiveness.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology and relates to a pharmaceutical composition, specifically a pharmaceutical composition of asiaticoside, leonurine and polygonatum polysaccharide, and its application in treating diseases such as diabetes and diabetic foot caused by diabetes, which have difficulty in wound healing. Background Technology

[0002] Diabetes mellitus is a metabolic disorder caused by genetic and environmental factors, leading to insulin insensitivity, insulin deficiency, and impaired biological function. Due to its high prevalence, associated disability, and mortality, diabetes has become a serious health problem worldwide.

[0003] Diabetes is classified into type 1 diabetes (insulin-dependent) and type 2 diabetes (non-insulin-dependent), with both genetic and environmental factors playing a role in its development. As an epidemic, diabetes has imposed an increasingly heavy social and economic burden due to its high morbidity and mortality rates. Prospective studies have shown that insulin resistance is one of the main pathogenic mechanisms of type 2 diabetes and also the best predictor, persisting throughout all stages of its development.

[0004] Diabetic complications are the most common complications known to exist, including diabetic nephropathy, diabetic retinopathy, and wound healing difficulties caused by hyperglycemia, such as diabetic foot. Once these complications occur, they are difficult to treat or reverse with medication. Diabetic foot is one of the most common complications of diabetes, characterized by slow wound healing and potential infection after foot injuries. The causes of this complication are varied, including diabetic vascular disease, neuropathy, and even infections. Diabetes can also cause impaired bone regeneration and repair, possibly related to metabolic disorders caused by long-term chronic hyperglycemia, impaired osteoblast proliferation and differentiation, and a high-glycemic environment in tissues that hinders wound repair, thus leading to complications such as diabetic foot.

[0005] Currently, there are many medications on the market for treating type 2 diabetes, mostly Western medicines such as sulfonylureas and biguanides. While these drugs are highly effective, their side effects are also significant, including bloating and diarrhea. There are currently no effective medications for treating diabetic foot and other diabetic injuries. Therefore, it is of great significance to develop a type 2 diabetes medication that reflects the holistic approach of traditional Chinese medicine, based on traditional Chinese medicine theory and utilizing modern science and technology, with clearly defined active ingredients, controllable quality, and minimal side effects. Summary of the Invention

[0006] The purpose of this invention is to provide a pharmaceutical composition of asiaticoside, leonurine, and polygonatum polysaccharide. This composition is obtained using modern scientific technology based on traditional Chinese medicine theory. It is simple to formulate, has a high content of active ingredients, and is safe and effective. Moreover, pharmacodynamic studies have shown that this pharmaceutical composition can lower blood sugar and promote wound healing, and is effective in treating diabetes and diabetic foot, as well as other diseases with poor wound healing.

[0007] Another object of the present invention is to provide the application of the above-mentioned pharmaceutical composition of asiaticoside, leonurine and polygonatum polysaccharide.

[0008] The objective of this invention can be achieved through the following technical solutions:

[0009] A pharmaceutical composition comprising asiaticoside, leonurine, and polygonatum polysaccharide, wherein the weight ratio of asiaticoside, leonurine, and polygonatum polysaccharide is 1:(1.2-2):(1-1.8).

[0010] Furthermore, the weight ratio of asiaticoside, leonurine, and polygonatum polysaccharide is 1:1.6:1.4.

[0011] Furthermore, in the pharmaceutical composition, the pharmaceutically acceptable carrier may take various forms. In the case of solid oral administration, suitable carriers include, but are not limited to: diluents, such as starch, corn starch, modified starch, lactose, powdered cellulose, microcrystalline cellulose, anhydrous calcium hydrogen phosphate, tricalcium phosphate, mannitol, sorbitol, sugar, maltodextrin, etc.; in addition, the carrier may also be sterile water, physiological saline or glucose aqueous solution; each carrier must be acceptable, compatible with other components in the pharmaceutical composition formulation and harmless to the patient.

[0012] Furthermore, the pharmaceutical composition can be formulated into a suitable pharmaceutical preparation form for oral or parenteral administration, including oral preparations such as tablets, capsules, and granules.

[0013] The use of a pharmaceutical composition of asiaticoside, leonurine and polygonatum polysaccharide in the preparation of a medicament for the prevention and / or treatment of diabetes-related diseases and / or conditions.

[0014] Furthermore, the disease and / or condition is selected from diabetes, diabetic foot caused by diabetes, or wound healing disorders caused by hyperglycemia.

[0015] Furthermore, the dosage of the pharmaceutical composition is 22.5-225 mg / day (based on a daily dosage volume of 750 ml).

[0016] Application of a pharmaceutical composition of asiaticoside, leonurine and polygonatum polysaccharide in the preparation of hypoglycemic drugs.

[0017] The formulation of this invention is based on the theory of traditional Chinese medicine, combined with modern research results, to further extract the active ingredients of Centella asiatica, Leonurus japonicus and Polygonatum sibiricum, and organically combine them, thereby developing a new Chinese medicine with high content of effective ingredients, controllable quality, few toxic side effects, and long-term use to treat diabetes and diseases such as diabetic foot caused by diabetes that are difficult to heal.

[0018] Centella asiatica has the effects of clearing heat and dampness, detoxifying and reducing swelling. In traditional Chinese medicine, it is generally used to treat damp-heat jaundice, heatstroke diarrhea, urinary stones and hematuria, carbuncles and boils, and injuries from falls. Asiaticoside is one of the effective components of Centella asiatica. Due to its wide range of pharmacological effects, it has a wide range of uses, most notably in clinical applications for repairing burn scars, and in skincare and cosmetic products as a skin-whitening, anti-inflammatory, and antioxidant ingredient. It may even have anti-tumor effects and enhance memory. Modern research shows that asiaticoside can lower blood sugar in type I and type II diabetic mice, maintain normal insulin secretion from pancreatic β-cells, increase HDL, and decrease TG. Regarding complications, asiaticoside can reduce urinary microalbumin, lower serum creatinine and blood urea nitrogen, and improve diabetic nephropathy. It has also been found to accelerate wound healing in diabetic foot mice, indicating a therapeutic effect on diabetic foot. Meanwhile, the study also found that when tumor-bearing mice were used to observe the effects of asiaticoside in combination with 5-fluorouracil (5-Fu) or cyclophosphamide (CTX) on the growth of human liver cancer cell line (Hep G2), asiaticoside could reduce the toxic side effects of chemotherapy drugs and prolong the lifespan of mice. This finding is of great significance for reducing the toxic side effects of drug combinations.

[0019] Motherwort can lower blood pressure, promote blood circulation and regulate menstruation, clear the liver and improve eyesight, nourish blood and dispel wind, and remove blood stasis and relieve pain. Leonuridine is an important active ingredient in motherwort. Modern research shows that it has good hypoglycemic and anti-osteoporosis effects. Animal experiments have confirmed that leonuridine can reduce fasting blood glucose in type II diabetic mice, improve oral glucose tolerance, and increase fasting plasma insulin. Furthermore, an explanation of the regulatory mechanism has been provided: leonuridine regulates the expression of glucokinase, glucose-6-phosphatase, and phosphoenolpyruvate carboxylase in a protein kinase B (Akt)-dependent manner. Therefore, leonuridine can be used as a drug to improve the symptoms of type II diabetes.

[0020] Polygonatum rhizome is a relatively precious traditional Chinese medicine. It tonifies qi and nourishes yin, strengthens the spleen, moistens the lungs, and benefits the kidneys. It can be used for symptoms such as spleen and stomach qi deficiency, fatigue, stomach yin deficiency, dry mouth and poor appetite, and dry cough due to lung deficiency. After screening dozens of hypoglycemic Chinese medicines, it was found that Polygonatum rhizome has the best hypoglycemic effect. Further using modern technology to separate the active components of Polygonatum rhizome, the hypoglycemic activity was identified as Polygonatum rhizome polysaccharide, which strongly inhibits α-glucosidase activity and has been used as a hypoglycemic drug with a significant hypoglycemic effect. Polygonatum rhizome polysaccharide, along with two purified components, PSP-1 and PSP-2, can effectively lower blood glucose in diabetic mice, regulate blood insulin levels, reduce insulin resistance, and significantly improve oral glucose tolerance in type II diabetic mice. Therefore, Polygonatum rhizome polysaccharide has important practical significance for the development of hypoglycemic Chinese medicines.

[0021] In summary, the formulation of asiaticoside, leonurine, and polygonatum polysaccharide is safe, effective, and complementary. It achieves significant hypoglycemic effects, promotes the treatment of diabetic foot, and has few toxic side effects. It holds promise as a long-term treatment option for diabetes and diabetic foot, as well as other conditions with difficult wound healing. Attached Figure Description

[0022] Figure 1 The effect of different concentrations of drug compositions in the same proportion on fasting blood glucose (FBG) in diabetic zebrafish;

[0023] Figure 2 The effect of different concentrations of drug compositions in the same proportion on the length of regenerated tail fins in diabetic zebrafish;

[0024] Figure 3 The effect of different proportions of the same concentration of drug composition on fasting blood glucose (FBG) in diabetic zebrafish;

[0025] Figure 4 The effect of different proportions of the same concentration of drug composition on the length of regenerated tail fin in diabetic zebrafish;

[0026] Among them, **P<0.001, ***P<0.0001, compared with the negative control group. Detailed Implementation

[0027] Unless otherwise stated, the terms used in this invention generally have the meanings commonly understood by those skilled in the art.

[0028] The present invention will be further described in detail below with reference to specific preparation examples and data. It should be understood that these examples are only for illustrating the present invention and are not intended to limit the scope of the present invention in any way.

[0029] In the following embodiments, various processes and methods not described in detail are conventional methods known in the art. The source, trade name, and components of the reagents used are indicated when they first appear. Unless otherwise specified, the same reagents used thereafter are the same as those indicated when they first appear.

[0030] The asiaticoside, leonurine, and polygonatum polysaccharide in this invention are all plant extracts or synthetic / semi-synthetic products.

[0031] The extraction methods for asiaticoside include, but are not limited to, decoction, maceration, percolation, reflux, microwave, ultrasonic, flash, and enzymatic extraction, with water, ethanol of different concentrations, and methanol as solvents.

[0032] Common extraction methods for Polygonatum polysaccharides include, but are not limited to, solvent extraction, flash extraction, microwave / ultrasound-assisted extraction, and enzyme extraction.

[0033] The extraction methods for leonurine include, but are not limited to, solvent reflux ultrasonic and microwave extraction; the synthesis of leonurine includes, but is not limited to: using syringic acid as a raw material, preparing a carbonylated product (4-ethoxycarbonyl-oxy-syringic acid-butyl ester) through reactions such as ethyl chloroformate protection and acylation of its active carbon group, ring opening of tetrahydrofuran under Lewis acid catalysis, and alcoholysis of acyl chloride; preparing leonurine using the Gabriel amine formation method; and finally reacting it with methyl isothiourea to produce leonurine. The obtained leonurine is, but is not limited to, leonurine, leonurine hydrochloride, or leonurine sulfate.

[0034] Example 1:

[0035] The preparation method of the pharmaceutical composition of asiaticoside, leonurine and polygonatum polysaccharide of the present invention is as follows: weigh 10g of asiaticoside, 12g of leonurine, and 10g of polygonatum polysaccharide, add appropriate excipients, granulate to prepare a preparation, and the preparation form is granules, tablets or capsules.

[0036] Example 2:

[0037] The preparation method of the pharmaceutical composition of asiaticoside, leonurine and polygonatum polysaccharide of the present invention is as follows: weigh 10g of asiaticoside, 16g of leonurine, and 14g of polygonatum polysaccharide, add appropriate excipients, granulate to prepare a preparation, and the preparation form is granules, tablets or capsules.

[0038] Example 3:

[0039] The preparation method of the pharmaceutical composition of asiaticoside, leonurine and polygonatum polysaccharide of the present invention is as follows: weigh 10g of asiaticoside, 20g of leonurine, and 18g of polygonatum polysaccharide, add appropriate excipients, granulate to prepare a preparation, and the preparation form is granules, tablets or capsules.

[0040] The pharmaceutical composition of asiaticoside, leonurine and polygonatum polysaccharide has the effects of treating diabetes, lowering blood sugar and treating difficult wound healing such as diabetic foot. These pharmacological effects have been confirmed by the following pharmacodynamic test examples.

[0041] I. Therapeutic effects of drug compositions of different concentrations and the same proportion on tail regeneration in adult diabetic zebrafish

[0042] Two hundred and forty adult zebrafish were randomly divided into four groups: a blank control group, a model group, a positive drug control group (Met-methyl), a 60 μg / mL drug composition administration group, a 120 μg / mL drug composition administration group, and a 240 μg / mL drug composition administration group, with 40 fish in each group. The weight ratio of asiaticoside, leonurine, and polygonatum polysaccharide in this experiment was 1:1.6:1.4 (i.e., the proportions of each component used in Example 2). All drugs were administered via immersion. Except for the blank group, all other groups were injected intraperitoneally with 0.3% streptozotocin to establish a diabetic model, while the blank group was injected with the same volume of citrate buffer. Blood glucose levels were measured after 3 days. After successful model establishment, all adult fish underwent tail amputation to establish a tail regeneration model for diabetic adult fish. After tail amputation, the blank group and the model group were placed in water, while the other groups were placed in their respective drug-containing culture media. Seven days later, blood glucose levels were tested in all groups, and the regenerated tail fins were observed under a microscope. After taking photos, the length of the regenerated tail fins was analyzed using software.

[0043] Blood glucose levels in the treatment group and the blank control group were statistically compared using the t-test. The results are shown in Table 1 and... Figure 1 .

[0044] Table 1. Effects of the drug composition on fasting blood glucose (FBG) levels in diabetic zebrafish.

[0045]

[0046] **P < 0.001, ***P < 0.0001, compared with the negative control group.

[0047] asiaticoside, leonurine, and polygonatum polysaccharide all have good hypoglycemic effects, and blood glucose level is an important indicator for measuring blood glucose levels. As shown in Table 1, the combination of the three drugs has a significant hypoglycemic effect, and the higher the concentration, the better the effect. The effect of the high-concentration drug combination even exceeds that of the positive control group.

[0048] The lengths of regenerated tail fins in the drug-treated group and the blank control group were statistically compared using a t-test. The results are shown in Table 2 and... Figure 2 .

[0049] Table 2. Effects of the drug composition on the length of regenerated tail fin in diabetic zebrafish.

[0050]

[0051] **P < 0.001, ***P < 0.0001, compared with the negative control group.

[0052] As shown in Table 2, the repair and regeneration speed of tail fin damage in the model group (diabetic fish) was slowed down due to the disease. The combination of the three drugs and the positive control drug could effectively promote the repair and regeneration speed of tail fin damage in the diseased fish. The medium and high concentration groups exceeded the positive control drug group, but the high concentration group promoted the tail fin regeneration speed less.

[0053] In summary, the decrease in blood glucose levels and the drug composition are dose-dependent, while the regeneration rate of the tail fin slows down with increasing concentration. Furthermore, angiogenesis can be observed in the low and medium concentration groups. Therefore, low and medium concentrations are more conducive to the repair and regeneration of zebrafish tail fin damage.

[0054] II. Therapeutic effects of drug compositions of different proportions and the same concentration on tail regeneration in adult diabetic zebrafish.

[0055] Two hundred and forty adult zebrafish were randomly divided into four groups: a blank control group, a model group, a positive drug control group (Met-methyl), a 1:1.2:1 drug composition group, a 1:1.6:1.4 drug composition group, and a 1:2:1.8 drug composition group, with 40 fish in each group. All drugs were administered via immersion. Except for the blank group, all other groups were injected intraperitoneally with 0.3% streptozotocin to establish a diabetic model. The blank group received the same volume of citrate buffer. Blood glucose levels were measured after three days. Once successful model establishment was confirmed, all adult fish underwent tail amputation to establish a tail regeneration model for diabetic adult fish. After tail amputation, the blank and model groups were placed in water, while the other groups were placed in their respective drug-containing culture media. Seven days later, blood glucose levels were measured in all groups, and the regenerated tail fins were observed under a microscope. The length of the regenerated tail fins was analyzed using software after photographing.

[0056] Blood glucose levels in the treatment group and the blank control group were statistically compared using t-tests. Results are shown in Table 3. Figure 3 .

[0057] Table 3. Effects of the drug composition on fasting blood glucose (FBG) levels in diabetic zebrafish.

[0058]

[0059] **P < 0.001, ***P < 0.0001, compared with the negative control group.

[0060] As shown in Table 3, all three ratios of the drug composition have significant hypoglycemic effects, and the hypoglycemic effects are similar at the same concentration.

[0061] The length of the regenerated tail fins in the drug-treated group and the blank control group were statistically compared using a t-test. The results are shown in Table 4 and... Figure 4 .

[0062] Table 4. Effects of the drug composition on the length of regenerated tail fin in diabetic zebrafish.

[0063]

[0064] **P < 0.001, ***P < 0.0001, compared with the negative control group.

[0065] As shown in Table 4, at the same concentration, the three drug compositions with different proportions all had a certain degree of promoting effect on the regeneration of zebrafish tail fins. However, the promoting effect slowed down when the proportion of leonurine and polygonatum polysaccharide became too high.

[0066] The pharmacodynamic tests above show that the combination of asiaticoside, leonurine, and polygonatum polysaccharide can effectively lower blood glucose levels and significantly promote the treatment of diabetic foot within a controllable concentration range. This indicates that the product has a significant therapeutic effect on diabetes and its complications, such as diabetic foot, and can be used to prepare drugs for the treatment of diabetes and its complications, such as diabetic foot, and for the repair of injuries (more specifically, the pharmaceutical composition of the present invention is suitable for lowering blood glucose in type 1 / type 2 diabetes and for treating type 1 / type 2 diabetes complicated with diabetic foot and symptoms such as impaired wound healing caused by hyperglycemia).

Claims

1. A pharmaceutical composition of asiaticoside, leonurine, and polygonatum polysaccharide, characterized in that, The pharmaceutical composition comprises asiaticoside, leonurine, and polygonatum polysaccharide, with a weight ratio of asiaticoside, leonurine, and polygonatum polysaccharide of 1:1.6:1.

4. The composition is used to prevent and / or treat diabetic foot caused by diabetes or wound healing disorders caused by hyperglycemia.

2. The pharmaceutical composition of asiaticoside, leonurine, and polygonatum polysaccharide as described in claim 1, characterized in that, The pharmaceutical composition can be formulated into capsules, tablets, or granules.

3. The use of the pharmaceutical composition of asiaticoside, leonurine and polygonatum polysaccharide as described in any one of claims 1-2 in the preparation of a medicament for the prevention and / or treatment of diabetic foot caused by diabetes or wound healing disorders caused by hyperglycemia.

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