Pharmaceutical composition for treating diabetic complications and application thereof

Abstract

The invention provides a pharmaceutical composition for treating diabetic complications and application thereof. Research finds that after effective parts such as coptis chinensis and radix puerariae are combined, the composition has obvious influence on diabetic complications and has a certain treatment effect. Specifically, the invention provides application of a coptis chinensis and kudzu vine root composition in preparation of a medicine for treating diabetic skin ulcer. The rhizoma coptidis and radix puerariae composition is prepared from the following raw materials in parts by weight: 1 to 10 parts of rhizoma coptidis alkaloid and 1 to 10 parts of rhizoma coptidis polysaccharide; the kudzuvine root extract is prepared from 1-10 parts by weight of kudzuvine root flavone, 1-10 parts by weight of kudzuvine root polysaccharide and 1-10 parts by weight of kudzuvine root saponin.

Description

Technical Field

[0001] This invention relates to the fields of traditional Chinese medicine and natural drug extracts. Background Technology

[0002] Diabetic complications refer to various complications that may occur after long-term diabetes, including cardiovascular disease, neuropathy, eye disease, and kidney disease. Because of the long-term instability of blood sugar levels, diabetic patients experience damage to various tissues and organs, leading to the development of complications.

[0003] Coptis chinensis, bitter and cold in nature, enters the heart, spleen, and stomach meridians. Its functions are clearing heat and drying dampness, purging fire and detoxifying. Traditionally used for "middle wasting disease" caused by excessive stomach fire. Its main component is coptis alkaloids. These can activate PKC, increase insulin receptor expression, repair damaged pancreatic islet cells, promote insulin secretion, reduce lipid deposition, regulate intestinal flora, and delay the hydrolysis and absorption of carbohydrates.

[0004] Kudzu root (Puerariae Lobatae Radix) is sweet, pungent, and cool in nature, and enters the spleen, stomach, and lung meridians. Its functions include relieving muscle tension and reducing fever, promoting body fluid production and quenching thirst. Traditionally, it is used for diabetes caused by yin deficiency and dryness-heat. Its main components are kudzu root flavonoids (or isoflavones). These can improve insulin resistance, protect pancreatic islet cells, and have anti-inflammatory and antioxidant effects.

[0005] The combination of these two ingredients is a core component of the classic formula Ge Gen Qin Lian Tang. This combination follows the treatment principle of "clearing heat and generating fluids." Ge Gen generates fluids and quenches thirst, treating the symptoms, while Huang Lian clears heat and purges fire, treating the root cause. It addresses both the symptoms and the root cause, and is especially suitable for treating type 2 diabetes with intestinal damp-heat or yin deficiency and dryness-heat.

[0006] There have been reports on the use of kudzu root and coptis root in combination to treat diabetes. Further research is needed to develop drug combinations that can effectively treat diabetic complications based on this. Summary of the Invention

[0007] In one aspect, the present invention provides a pharmaceutical composition for treating diabetic complications, comprising the following components:

[0008] (1) 1-10 parts by weight of Coptis chinensis alkaloids;

[0009] (2) 1-10 parts by weight of kudzu flavonoids, 1-10 parts by weight of kudzu polysaccharides, or / and 1-10 parts by weight of kudzu saponins.

[0010] In one aspect, the present invention also provides a pharmaceutical composition for treating diabetic complications, characterized in that it comprises the following components:

[0011] (1) 1-10 parts by weight of Coptis chinensis polysaccharide;

[0012] (2) 1-10 parts by weight of kudzu flavonoids, 1-10 parts by weight of kudzu polysaccharides, or / and 1-10 parts by weight of kudzu saponins.

[0013] In one aspect, the present invention also provides a pharmaceutical composition for treating diabetic complications, comprising the following components:

[0014] (1) 1-10 parts by weight of berberine;

[0015] (2) 1-10 parts by weight of kudzu flavonoids, 1-10 parts by weight of kudzu polysaccharides, or / and 1-10 parts by weight of kudzu saponins.

[0016] In one aspect, the present invention also provides a pharmaceutical composition for treating diabetic complications, comprising the following components:

[0017] (1) 1-10 parts by weight of kudzu flavonoids;

[0018] (2) 1-10 parts by weight of Coptis chinensis alkaloids, 1-10 parts by weight of Coptis chinensis polysaccharides, or / and 1-10 parts by weight of Coptis chinensis flavonoids.

[0019] In one aspect, the present invention also provides a pharmaceutical composition for treating diabetic complications, comprising the following components:

[0020] (1) 1-10 parts by weight of kudzu polysaccharide;

[0021] (2) 1-10 parts by weight of Coptis chinensis alkaloids, 1-10 parts by weight of Coptis chinensis polysaccharides, or / and 1-10 parts by weight of Coptis chinensis flavonoids.

[0022] In one aspect, the present invention also provides a pharmaceutical composition for treating diabetic complications, comprising the following components:

[0023] (1) 1-10 parts by weight of kudzu saponins;

[0024] (2) 1-10 parts by weight of Coptis chinensis alkaloids, 1-10 parts by weight of Coptis chinensis polysaccharides, or / and 1-10 parts by weight of Coptis chinensis flavonoids.

[0025] In one aspect, the present invention also provides a pharmaceutical composition for treating diabetic complications, comprising the following components:

[0026] (1) 1-10 parts by weight of Coptis chinensis alkaloids and 1-10 parts by weight of Coptis chinensis polysaccharides;

[0027] (2) 1-10 parts by weight of kudzu flavonoids and 1-10 parts by weight of kudzu polysaccharides; or 1-10 parts by weight of kudzu flavonoids and 1-10 parts by weight of kudzu saponins; or 1-10 parts by weight of kudzu polysaccharides and 1-10 parts by weight of kudzu saponins.

[0028] In one aspect, the present invention also provides a pharmaceutical composition for treating diabetic complications, comprising the following components:

[0029] (1) 1-10 parts by weight of Coptis chinensis alkaloids and 1-10 parts by weight of Coptis chinensis flavonoids;

[0030] (2) 1-10 parts by weight of kudzu flavonoids and 1-10 parts by weight of kudzu polysaccharides; or 1-10 parts by weight of kudzu flavonoids and 1-10 parts by weight of kudzu saponins; or 1-10 parts by weight of kudzu polysaccharides and 1-10 parts by weight of kudzu saponins.

[0031] In one aspect, the present invention also provides a pharmaceutical composition for treating diabetic complications, comprising the following components:

[0032] (1) 1-10 parts by weight of Coptis polysaccharide and 1-10 parts by weight of Coptis flavonoids;

[0033] (2) 1-10 parts by weight of kudzu flavonoids and 1-10 parts by weight of kudzu polysaccharides; or 1-10 parts by weight of kudzu flavonoids and 1-10 parts by weight of kudzu saponins; or 1-10 parts by weight of kudzu polysaccharides and 1-10 parts by weight of kudzu saponins.

[0034] In one aspect, the present invention also provides a pharmaceutical composition for treating diabetic complications, comprising the following components:

[0035] (1) 1-10 parts by weight of Coptis chinensis alkaloids, 1-10 parts by weight of Coptis chinensis polysaccharides, and 1-10 parts by weight of Coptis chinensis flavonoids;

[0036] (2) 1-10 parts by weight of kudzu flavonoids, 1-10 parts by weight of kudzu polysaccharides, and 1-10 parts by weight of kudzu saponins.

[0037] The amount of each component is independently selected from 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 parts by weight.

[0038] The present invention also provides the use of the above-described pharmaceutical composition in the preparation of a medicament for treating diabetic complications.

[0039] Diabetic skin ulcers, also known as diabetic skin sores, are ulcers caused by the hyperglycemic state resulting from diabetes. This hyperglycemia leads to microvascular and neuropathy in the skin, causing ischemia, necrosis, and infection of the skin tissue, resulting in ulcers that are difficult to heal. These ulcers typically occur on the feet and lower extremities, but can also appear on other sites such as the hands and abdomen. Diabetic skin sores not only affect a patient's quality of life but can also lead to serious infections and the risk of amputation.

[0040] The present invention also provides the use of the Coptis chinensis and Pueraria lobata composition in the preparation of a medicament for treating diabetic skin ulcers;

[0041] The Coptis chinensis and Pueraria lobata composition is composed of the following raw materials in parts by weight:

[0042] Coptis chinensis extract: Pueraria lobata extract = 2~4: 7~9;

[0043] Among them, the Coptis chinensis extract consists of 1-10 parts by weight of Coptis chinensis alkaloids and 1-10 parts by weight of Coptis chinensis polysaccharides; the Pueraria lobata extract consists of 1-10 parts by weight of Pueraria lobata flavonoids, 1-10 parts by weight of Pueraria lobata polysaccharides and 1-10 parts by weight of Pueraria lobata saponins.

[0044] In some specific embodiments of the present invention, the ratio of Coptis chinensis extract to Pueraria lobata extract is 3:8.

[0045] In some specific embodiments of the present invention, the Coptis chinensis extract consists of 1 part by weight of Coptis chinensis alkaloids and 1 part by weight of Coptis chinensis polysaccharides.

[0046] In some specific embodiments of the present invention, the kudzu root extract consists of 1 part by weight of kudzu flavonoids, 1 part by weight of kudzu polysaccharides, and 1 part by weight of kudzu saponins.

[0047] In some specific embodiments of the present invention, the treatment of diabetic skin ulcers is to promote the healing of diabetic skin ulcer wounds.

[0048] In some specific embodiments of the present invention, a pharmaceutical composition for treating diabetic complications comprises the following raw materials in parts by weight:

[0049] Coptis chinensis extract: Pueraria lobata extract = 2~4: 7~9;

[0050] Among them, the Coptis chinensis extract consists of 1-10 parts by weight of Coptis chinensis alkaloids and 1-10 parts by weight of Coptis chinensis polysaccharides; the Pueraria lobata extract consists of 1-10 parts by weight of Pueraria lobata flavonoids, 1-10 parts by weight of Pueraria lobata polysaccharides and 1-10 parts by weight of Pueraria lobata saponins.

[0051] In some specific embodiments of the present invention, the ratio of Coptis chinensis extract to Pueraria lobata extract is 3:8.

[0052] In some specific embodiments of the present invention, the Coptis chinensis extract consists of 1 part by weight of Coptis chinensis alkaloids and 1 part by weight of Coptis chinensis polysaccharides.

[0053] In some specific embodiments of the present invention, the kudzu root extract consists of 1 part by weight of kudzu flavonoids, 1 part by weight of kudzu polysaccharides, and 1 part by weight of kudzu saponins.

[0054] This invention has verified that the combination of the above-mentioned effective components such as Coptis chinensis and Pueraria lobata has a significant effect on diabetic skin ulcers, which are a complication of diabetes, and has a certain therapeutic effect.

[0055] The advantages of this invention are as follows:

[0056] 1. Filling the gap in existing technology: Although some components of Coptis chinensis and Pueraria lobata (such as alkaloids and flavonoids) have been studied for use in diabetes, or individual components (such as Coptis chinensis polysaccharides) have been studied for use in diabetic wounds, there has never been any existing technology that systematically combines the five active components of the "Coptis chinensis-Pueraria lobata" system ("Coptis chinensis alkaloids, Coptis chinensis polysaccharides, Pueraria lobata flavonoids, Pueraria lobata polysaccharides, and Pueraria lobata saponins") and specifically uses them for the treatment of diabetic skin ulcers.

[0057] 2. Synergistic Innovation with Multiple Components and Targets: It is well known that diabetic skin ulcers are chronic and difficult to heal due to multiple factors such as a high-sugar environment, susceptibility to infection, and microcirculatory disturbances, resulting in a lack of highly effective clinical treatments. This invention addresses the complex pathology of this disease with a refined design, employing specific "Coptis chinensis-Pueraria lobata" species-derived active ingredients. These components work together to exert powerful antibacterial and anti-inflammatory effects, improve microcirculation, promote angiogenesis and tissue repair, producing a synergistic effect of '1+1>2' in promoting the healing of diabetic skin ulcers. Attached Figure Description

[0058] Figure 1 Photo of experimental animals Detailed Implementation

[0059] The berberine, berberine polysaccharide, kudzu flavonoids, kudzu polysaccharide, and kudzu saponins described in this invention can all be extracted and purified using existing technologies, or obtained by purchasing existing products.

[0060] Alkaloids can be extracted and purified using conventional methods: ethanol reflux extraction, after removing the ethanol from the extract, loading it onto a macroporous resin column, optimizing parameters such as the diameter-to-height ratio, sample concentration and pH, and eluent (ethanol) concentration and volume, and tracking the elution by methods such as TLC or HPLC to collect the eluent components with higher content, thus obtaining alkaloids.

[0061] Polysaccharides can be extracted and purified using conventional methods: hot water extraction or enzyme-assisted hot water extraction, followed by impurity removal (deproteinization / decolorization), and then elution with water using macroporous adsorption resin to obtain the polysaccharide. To achieve the target purity, further purification can be performed using ion exchange resins or gel column chromatography.

[0062] Flavonoids can be extracted and purified using conventional methods: hot reflux extraction with ethanol / alcohol solvent, purification with an appropriate concentration of aqueous ethanol in macroporous resin, generally using D101 or AB-8, to obtain the flavonoid fraction.

[0063] Saponins can be extracted and purified using conventional methods: hot reflux extraction with ethanol / alcohol solvent, followed by extraction with n-butanol, or / and purification with an appropriate concentration of aqueous ethanol using a macroporous resin column to obtain the saponin fraction.

[0064] In each extract of the present invention, the labeled amount of the corresponding substance should reach more than 20%.

[0065] Kudzu polysaccharide was prepared by the following method: Zhu Jiaqing, et al. Study on purification process and antioxidant properties of kudzu polysaccharide [J]. Food Industry Technology, 2020, 41(24):131-136.

[0066] Pueraria saponins were prepared by the following method: Wang Zhiping, et al. Study on purification of total pueraria saponins by macroporous adsorption resin [J]. Journal of Guangdong Pharmaceutical University, 2012, 28(02):145-148.

[0067] Kudzu flavonoids were prepared by the following method: Xu Yan. Isolation and purification of chemical and bioactive substances from kudzu root [D]. Anhui Agricultural University, 2003.

[0068] Coptis chinensis polysaccharides were prepared by the following method: Yin Dangyue. Research on the full resource utilization processing of Coptis chinensis and the bioactivity of its products [D]. Northwest A&F University, 2022.

[0069] Coptis chinensis alkaloids were prepared by the following method: He Zhilong. Preliminary study on the composition and preparation of total isoflavones from kudzu root and total alkaloids from coptis chinensis [D]. Southwest Jiaotong University, 2015.

[0070] Example 1

[0071] Take 1g of kudzu saponin, 1g of kudzu flavonoids, 1g of kudzu polysaccharide, 1g of coptis alkaloids, and 1g of coptis polysaccharide, and mix them to obtain the composition.

[0072] Example 2

[0073] Take 8g of kudzu saponins, 8g of kudzu flavonoids, 8g of kudzu polysaccharides, 3g of coptis alkaloids, and 3g of coptis polysaccharides, and mix them to obtain the composition.

[0074] Example 3

[0075] Take 9g of kudzu saponins, 9g of kudzu flavonoids, 9g of kudzu polysaccharides, 2g of coptis alkaloids, and 2g of coptis polysaccharides, and mix them to obtain the composition.

[0076] Example 4

[0077] Take 7g of kudzu saponins, 7g of kudzu flavonoids, 7g of kudzu polysaccharides, 4g of coptis alkaloids, and 4g of coptis polysaccharides, and mix them to obtain the composition.

[0078] Coptis chinensis alkaloids

[0079] Definition: Berberine alkaloids are a class of alkaloids extracted from the dried rhizomes of the Ranunculaceae plant Coptis chinensis Franch. The main components include berberine, palmatine, and berberine. These alkaloids possess antiviral, anti-inflammatory, antibacterial, antitumor, hypoglycemic, and immunomodulatory effects, and are widely used in food, health care, and pharmaceutical fields.

[0080] Standard: The quality standard of Coptis chinensis extract shall comply with the provisions of Part I and Part IV of the Chinese Pharmacopoeia.

[0081] Coptis polysaccharide

[0082] Definition: Coptis chinensis polysaccharides are a class of polysaccharides extracted from Coptis chinensis, possessing various biological activities such as immunomodulation, antitumor activity, and antioxidant activity. The extraction and purification processes of Coptis chinensis polysaccharides are relatively complex, but they have significant application value in the pharmaceutical and health care fields.

[0083] Kudzu polysaccharides

[0084] Definition: Kudzu polysaccharides are a class of polysaccharides extracted from the roots of the legume kudzu (Pueraria lobata (Willd.) Ohwi), which have various biological activities such as immunomodulation, anti-inflammation, and hypoglycemia.

[0085] Pueraria Flavonoids

[0086] Definition: Kudzu flavonoids are a class of flavonoid compounds extracted from kudzu root, with main components including puerarin and daidzein. Kudzu flavonoids possess various pharmacological effects, including vasodilatory activity, improved microcirculation, anti-inflammatory properties, and antioxidant activity, and are widely used in traditional Chinese medicine preparations, food additives, and health products.

[0087] Puerarin saponins

[0088] Definition: Puerarin saponins are a class of saponin compounds extracted from kudzu root. They possess various pharmacological effects, such as anti-inflammatory, antiviral, and hypoglycemic properties. Although the content of kudzu saponins in kudzu root is relatively low, they exhibit significant biological activity.

[0089] This invention has verified that the combination of the above-mentioned effective components such as Coptis chinensis and Pueraria lobata has a significant effect on diabetic complications and has a certain therapeutic effect.

[0090] Experimental Example: Treatment of Diabetic Skin Ulcers with the Composition of the Invention

[0091] I. Experimental Design

[0092] 1. Experimental Grouping

[0093] Blank group, model group, and Coptis chinensis and Pueraria lobata composition group (prepared in Example 2).

[0094] 2. Preparation of a diabetes model

[0095] Healthy male SD rats were fasted for 12 hours before modeling. Streptozotocin (STZ) was prepared using 0.5 mmol / L citrate buffer (pH 4.4-5.6) (1 ml citrate buffer was added for every 0.207 g STZ). The rats were induced to develop diabetes by intraperitoneal injection at a dose of 80 mg / kg. After 72 hours, blood glucose was measured by tail vein sampling for three consecutive days. If the blood glucose concentration was ≥16.7 mmol / L and the rats exhibited symptoms such as polydipsia, polyphagia, and polyuria, the diabetes model was considered to have been successfully established. No intervention was given to the normal group rats.

[0096] 3. Preparation of a diabetic skin ulcer model

[0097] (Skin modeling was performed three days after STZ injection.) Diabetic rats and normal rats with successfully established models were anesthetized by intraperitoneal injection of chloral hydrate at 350 mg / kg. The backs were shaved to create bare skin areas, which were then depilated with hair removal cream. The bare skin on the rat's back was disinfected with povidone-iodine. A circular piece of full-thickness skin tissue, approximately 1.5 cm × 1.5 cm in size, extending to the fascia layer, was removed using a punch, thus successfully establishing a diabetic rat skin ulcer model.

[0098] 4. Administration method

[0099] In the normal group and the model group, the wound was cleaned with physiological saline, and 3M dressing was applied to the wound and fixed with adhesive tape. This was done once every other day for 14 consecutive days. In the treatment group (composition 1100mg / Kg, dissolved in UP water, 2ml per rat by gavage), the wound was cleaned with physiological saline, and the drug was administered by gavage. The wound was then covered with sterile gauze and fixed with adhesive tape. This was done once every other day for 14 consecutive days.

[0100] 5. Draw materials

[0101] After the last administration, rats in each group were fasted for 12 hours but allowed free access to water. Blood was collected from the abdominal aorta after intraperitoneal anesthesia with 10% chloral hydrate. Serum was collected by centrifugation and frozen for later use. Wound tissue and wound margin tissue were harvested. The wound was divided into four quadrants. The left side was divided into upper and lower sections and fixed in 4% paraformaldehyde for storage at room temperature. The right side was also divided into upper and lower sections, and cryopreserved in cryovials at -80°C after flash freezing in liquid nitrogen.

[0102] 6. Indicator Testing

[0103] The wound healing status of rats in each group after drug intervention was recorded by photograph, and the changes in wound color, shrinkage, and presence of exudate were observed. ImageJ software was used to calculate the wound area recorded on copper sulfate paper at 3, 7, and 14 days after drug administration, and the wound healing rate was calculated.

[0104] Calculation formula:

[0105] Wound healing rate (%) = [(Initial ulcer area - Unhealed ulcer area) / Initial ulcer area] × 100%.

[0106] Anti-inflammatory properties assay: In vivo anti-inflammatory performance was detected by immunofluorescence staining.

[0107] Histological observation: HE staining, Messin staining;

[0108] Measurement of oxidative stress indicators: Centrifugation was used to collect serum samples to measure the activity or content of SOD, MDA, and GSH.

[0109] Inflammatory response assay: ELISA was used to measure the levels of IL-1β, IL-6, TNF-α, and VEGF;

[0110] RT-qPCR was used to measure the expression of HIF-1α and VEGFA (some angiogenesis-related growth factors) in wound tissue. Immunohistochemistry was used to measure the protein expression of HIF-1α and VEGFA in the wound.

[0111] II. Experimental Results

[0112] 1. Blood glucose level

[0113] Blood glucose was measured randomly once a week, and the random blood glucose concentration was higher than 16.7 mmol / L, meaning that all animals after modeling suffered from hyperglycemia from the beginning to the end of the experiment.

[0114] 2. Ulcer outcome

[0115] Three animals from each experimental group were selected for wound image comparison. The results showed that the healing rate in the drug-treated group was higher than that in the model group, and lower than or equal to that in the control group. The healing rate in the control group was higher than that in the model group. See details... Figure 1 .

Claims

1. Use of the Coptis chinensis and Pueraria lobata composition in the preparation of a medicament for treating diabetic skin ulcers; The Coptis chinensis and Pueraria lobata composition is composed of the following raw materials in parts by weight: Coptis chinensis alkaloids 1-10 parts by weight, Coptis chinensis polysaccharides 1-10 parts by weight; Pueraria lobata extract contains Pueraria lobata flavonoids 1-10 parts by weight, Pueraria lobata polysaccharides 1-10 parts by weight, and Pueraria lobata saponins 1-10 parts by weight.

2. The use according to claim 1, characterized in that, The extract of Coptis chinensis consists of 2-4 parts by weight of Coptis chinensis alkaloids and 2-4 parts by weight of Coptis chinensis polysaccharides.

3. The use according to claim 1, characterized in that, The kudzu root extract contains 7-9 parts by weight of kudzu flavonoids, 7-9 parts by weight of kudzu polysaccharides, and 7-9 parts by weight of kudzu saponins.

4. The use according to claim 1, characterized in that, The Coptis chinensis extract consists of 3 parts by weight of Coptis chinensis alkaloids and 3 parts by weight of Coptis chinensis polysaccharides; the Pueraria lobata extract consists of 8 parts by weight of Pueraria lobata flavonoids, 8 parts by weight of Pueraria lobata polysaccharides, and 8 parts by weight of Pueraria lobata saponins.

5. The use according to claim 1, characterized in that, The treatment for diabetic skin ulcers aims to promote the healing of diabetic skin ulcer wounds.

6. A pharmaceutical composition for treating diabetic complications, characterized in that, It consists of the following raw materials in parts by weight: Coptis chinensis alkaloids 1-10 parts by weight, Coptis chinensis polysaccharides 1-10 parts by weight; Pueraria lobata extract contains Pueraria lobata flavonoids 1-10 parts by weight, Pueraria lobata polysaccharides 1-10 parts by weight, and Pueraria lobata saponins 1-10 parts by weight.

7. The pharmaceutical composition according to claim 6, characterized in that, The extract of Coptis chinensis consists of 2-4 parts by weight of Coptis chinensis alkaloids and 2-4 parts by weight of Coptis chinensis polysaccharides.

8. The pharmaceutical composition according to claim 6, characterized in that, The kudzu root extract contains 7-9 parts by weight of kudzu flavonoids, 7-9 parts by weight of kudzu polysaccharides, and 7-9 parts by weight of kudzu saponins.

9. The pharmaceutical composition according to claim 6, characterized in that, The Coptis chinensis extract consists of 3 parts by weight of Coptis chinensis alkaloids and 3 parts by weight of Coptis chinensis polysaccharides; the Pueraria lobata extract consists of 8 parts by weight of Pueraria lobata flavonoids, 8 parts by weight of Pueraria lobata polysaccharides, and 8 parts by weight of Pueraria lobata saponins.

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