A pharmaceutical composition for treating peripheral neuropathy of type 2 diabetes mellitus, preparation and application thereof

Abstract

The application discloses a kind of drug composition for treating type 2 diabetes peripheral neuropathy and its preparation and application, the drug composition is composed of Bai Cao, Diandongolivae, Diandongjiuxueteng and hirudo. The drug composition adds pharmaceutically acceptable adjuvant, and is made into preparation. The drug composition and its preparation can be used to prepare the drug for treating type 2 diabetes peripheral neuropathy.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceuticals, specifically relating to a pharmaceutical composition for treating type 2 diabetic peripheral neuropathy, its formulation, and its application. Background Technology

[0002] Type 2 diabetic peripheral neuropathy (T2DPN) is one of the most common and serious microvascular complications of type 2 diabetes mellitus (T2DM). According to the International Diabetes Federation (IDF), the global prevalence of diabetes has reached 11.2%, with approximately 50% of patients eventually developing DPN. This disease primarily affects the peripheral, central, and autonomic nervous systems, with clinical manifestations mainly including symmetrical numbness, pain, paresthesia, or loss of sensation in the distal extremities. In severe cases, it can lead to lower extremity ulcers and even amputation, significantly increasing patient disability and mortality rates and imposing a heavy economic burden on society and families.

[0003] The pathogenesis of type 2 diabetic neuropathy (T2DPN) is not yet fully understood, but it is generally believed to be related to multiple factors, including hyperglycemia-induced metabolic disorders, oxidative stress, microvascular damage, and a lack of neurotrophic factors. In terms of treatment, modern medicine mainly employs symptomatic treatments such as controlling blood sugar, nourishing nerves (e.g., methylcobalamin), providing antioxidants (e.g., lipoic acid), and improving microcirculation. However, existing treatments have significant limitations: first, there is a lack of drugs that specifically block the progression of neuropathy; most drugs can only relieve symptoms and cannot reverse pathological changes; second, long-term use may be accompanied by dose-dependent side effects; and third, symptoms are prone to relapse after discontinuation of medication, resulting in a bottleneck in overall clinical efficacy. Given the limitations of existing treatments, leveraging the holistic concept and syndrome differentiation of traditional Chinese medicine (TCM), developing a multi-component, multi-target, and multi-pathway TCM compound or comprehensive treatment plan that can effectively improve limb numbness and pain caused by Qi and Yin deficiency and damp-heat stagnation, with a low recurrence rate and high safety, has become an urgent need in current clinical research. Summary of the Invention

[0004] To address the shortcomings of existing technologies, the present invention aims to provide a pharmaceutical composition, its formulation, and its application for treating type 2 diabetic peripheral neuropathy. The pharmaceutical composition of the present invention is effective in treating type 2 diabetic peripheral neuropathy and is safe with no toxic side effects.

[0005] The objective of this invention is achieved through the following technical solution: A pharmaceutical composition for treating type 2 diabetic peripheral neuropathy, comprising, by weight, the following components: 10-40 parts of Agrimonia pilosa, 5-20 parts of Oligospermum jasminoides, 5-20 parts of Spatholobus suberectus, and 2-9 parts of Hirudo medicinalis.

[0006] Preferably, the pharmaceutical composition for treating type 2 diabetic peripheral neuropathy comprises, by weight, the following components: 15-35 parts of Agrimonia pilosa, 10-20 parts of Oligospermum jasminoides, 10-20 parts of Spatholobus suberectus, and 3-8 parts of Hirudo medicinalis.

[0007] More preferably, the pharmaceutical composition for treating type 2 diabetic peripheral neuropathy comprises, by weight, the following components: 20-35 parts of Agrimonia pilosa, 10-15 parts of Oligospermum jasminoides, 10-15 parts of Spatholobus suberectus, and 4-6 parts of Hirudo medicinalis.

[0008] Traditional Chinese medicine has a profound understanding of this disease. Although it does not have the name "diabetic peripheral neuropathy," its clinical manifestations can be classified under the categories of "Xiao Ke" (wasting and thirsting disease), "Bi Zheng" (bi syndrome), "Wei Zheng" (atrophy syndrome), or "Nu Mu" (numbness) in traditional Chinese medicine. The "Secret Transmission of Essential Treatments" also states: "If Xiao Ke has been present for a long time... it may cause hemiplegia and numbness of the hands and feet, resembling a stroke," which vividly describes the evolution of neuropathy as a complication of Xiao Ke.

[0009] Traditional Chinese medicine theory holds that type 2 diabetes mellitus (T2DPN) often occurs secondary to prolonged diabetes mellitus, with its core pathogenesis being deficiency of both qi and yin. Prolonged diabetes mellitus depletes body fluids, leading to yin deficiency and dryness. Over time, qi follows yin depletion, resulting in insufficient qi to propel blood circulation and insufficient yin to nourish the tendons and vessels. Furthermore, deficiency of both qi and yin easily leads to impaired metabolism, generating internal damp-heat. This damp-heat, combined with blood stasis, obstructs the meridians, causing malnourishment of the extremities. "Where there is no flow, there is pain; where there is no nourishment, there is numbness," ultimately resulting in numbness and pain in the limbs.

[0010] The treatment should focus on clearing heat and drying dampness, promoting blood circulation and unblocking collaterals, and generating fluids and moistening dryness. The principal herb, *Potentilla discolor*, is sweet and slightly bitter, neutral to slightly cold in nature, and enters the stomach and large intestine meridians. It has the effects of cooling the blood and stopping bleeding, clearing heat and detoxifying, and can clear the stomach and intestines. Its bitter and cold nature can directly clear damp heat from the body, especially for cases of damp heat descending and infiltrating the meridians, clearing the damp heat steaming in the meridians and relieving burning, numbness, and pain in the limbs. Its sweet and slightly cold nature can also promote fluid production, avoiding the damage to fluids caused by bitter cold, which is suitable for the underlying yin deficiency of long-term diabetes, eliminating damp heat without depleting body fluids. The assistant herb, *Oliveum yunnanense*, is sweet and sour, cool in nature, and enters the liver, lung, spleen, and stomach meridians. Its effects include generating fluids and quenching thirst, clearing heat and relieving sore throat, cooling the blood, and promoting digestion and strengthening the stomach. It assists the principal herb in enhancing its heat-clearing power, and its cooling and moistening nature can restrain the dryness and intensity of damp heat. Its sweet and sour nature can generate fluids and quench thirst, nourish yin fluids, and restored body fluids can nourish the meridians. When used in combination, the principal and assistant herbs work synergistically to clear heat and generate fluids, ensuring that "damp-heat is cleared without depleting fluids, and fluids are generated without exacerbating dampness." The adjuvant herb, *Spatholobus suberectus*, is bitter and sweet in taste, warm in nature, and enters the liver and kidney meridians. Its functions include invigorating blood circulation, replenishing blood, and relaxing muscles and tendons. Its blood-invigorating power can unblock the meridians and resolve blood stasis; its blood-replenishing function addresses the latent deficiency of qi and blood in long-term diabetes, ensuring that invigorating blood circulation does not damage it, and that unblocking the meridians while nourishing the body. This promotes smooth qi and blood flow, improves blood circulation in the limbs, and relieves symptoms such as numbness and pain. Its warm and dispersing nature allows it to guide the medicinal power of the principal and assistant herbs directly to the meridians of the limbs, especially focusing on the peripheral nerves, enhancing the local effects of clearing heat, generating fluids, and unblocking the meridians. As an adjuvant herb, it also acts as a guiding herb, connecting the upper and lower parts of the body and penetrating the interior and exterior. Leeches are salty and bitter in taste, neutral in nature, and enter the liver meridian. Their effects include breaking up blood stasis, clearing the meridians, and eliminating masses. The blood stasis in diabetic peripheral neuropathy is often stubborn blood stasis that has "entered the collaterals due to prolonged illness." Ordinary blood-activating drugs are difficult to achieve results. Leeches have a strong ability to break up blood stasis and can penetrate the meridian obstruction to clear the blood vessels, making them a key drug for clearing the collaterals. The formula takes advantage of its ability to break up blood stasis and clear the collaterals while avoiding its strong effect that may harm the body's vital energy, which is in line with the principle of "stopping when the disease is cured and protecting the body's vital energy." When combined with Spatholobus suberectus, the two are one strong and one mild, one breaking up blood stasis and the other replenishing blood. Leeches break up blood stasis and clear the collaterals, while Spatholobus suberectus replenishes blood and activates blood circulation. They complement each other, so that blood stasis can be quickly removed without harming the body's vital energy. When used together, these herbs work synergistically to clear heat and dry dampness, generate fluids and moisten dryness, invigorate blood and remove blood stasis, and unblock the meridians and relieve pain. This is consistent with the syndrome of diabetic peripheral neuropathy characterized by internal damp-heat and blood stasis obstructing the meridians, thus clearing damp-heat, dispersing blood stasis, restoring body fluids, and unblocking the meridians, thereby eliminating various symptoms in the limbs.

[0011] The pharmaceutical composition for treating type 2 diabetic peripheral neuropathy may also be formulated with pharmaceutically acceptable excipients to produce granules, oral liquids, mixtures, tablets, powders, pills, concentrated pills, sustained-release microcapsules, and other preparations.

[0012] The pharmaceutical composition and formulation thereof for treating type 2 diabetic peripheral neuropathy can be used to prepare a drug for treating type 2 diabetic peripheral neuropathy.

[0013] Compared with the prior art, the present invention has the following beneficial effects: 1. The pharmaceutical composition of the present invention is formulated with Yunnan ethnic medicines. Clinical use has shown that it has good efficacy and no adverse reactions have been observed.

[0014] 2. The pharmaceutical composition of this invention can effectively improve the symptoms of peripheral neuropathy and related pathological indicators in a type 2 diabetic rat model. The pharmaceutical composition can significantly reduce the body weight and fasting blood glucose level of the model rats, improve oral glucose tolerance, and reduce insulin resistance; it can also regulate lipid metabolism disorders and reduce serum total cholesterol, triglycerides, and low-density lipoprotein cholesterol levels. Furthermore, the pharmaceutical composition also has anti-inflammatory effects, significantly reducing the levels of pro-inflammatory factors (IL-1β, IL-6, TNF-α) and increasing the levels of anti-inflammatory factors (IL-10). Regarding neurological function, the pharmaceutical composition can improve the microcirculation of the sciatic nerve tissue, increase nerve blood flow and vascular density, and enhance the conduction velocity of motor and sensory nerves; it can also reduce the content of malondialdehyde (MDA) in the sciatic nerve tissue and increase superoxide dismutase (SOD), glutathione peroxidase (GSH-PX), and sodium-potassium pump (Na+) levels. + -K + The presence of ATPase activity indicates that it can alleviate oxidative stress damage and improve neuronal energy metabolism. These results demonstrate that the pharmaceutical composition of this invention exerts a therapeutic effect on type 2 diabetic peripheral neuropathy by comprehensively regulating multiple pathological processes, including glucose and lipid metabolism, inflammatory response, oxidative stress, and neural microcirculation.

[0015] 3. The pharmaceutical composition described in this invention has significant clinical efficacy in treating type 2 diabetic peripheral neuropathy. This drug can significantly improve patients' TCM syndromes, effectively alleviating symptoms such as limb numbness, limb pain, fatigue, dry mouth and throat, hot flashes in the palms, soles, and chest, dull complexion, and dark lips and nails; simultaneously, it significantly reduces body weight and BMI, regulates glucose and lipid metabolism disorders, and improves insulin resistance. Furthermore, by reducing whole blood and plasma viscosity, improving blood rheology and microcirculation, this drug promotes the repair and functional recovery of peripheral nerve structures, increases nerve conduction velocity, and significantly relieves symptoms such as limb numbness and pain, ultimately achieving holistic intervention and effective treatment for type 2 diabetic peripheral neuropathy.

[0016] 4. The pharmaceutical composition of the present invention can also be prepared as a formulation loaded with Ni, Mo, N co-doped mesoporous carbon material (Mo,Ni-N-MCS). Berberine and astragaloside are the characteristic active ingredients of the pharmaceutical composition of the present invention. This mesoporous material has a regular pore structure and good biocompatibility, which can effectively load and sustain the release of active pharmaceutical ingredients (such as berberine and astragaloside), achieving long-term drug delivery. At the same time, its conductivity can provide physical support for nerve repair, synergistically with the chemical regulation of the drug, and jointly promote nerve regeneration and functional recovery in diabetic peripheral neuropathy from both the aspects of "physical support repair" and "chemical regulation of the microenvironment". Attached Figure Description

[0017] Figure 1 This is a SEM image of the N-doped mesoporous material (N-MCS) prepared in Example 15.

[0018] Figure 2 The image shows a SEM image of the Mo,Ni-N-MCS prepared in Example 15.

[0019] Figure 3 This is a TEM image of the N-MCS prepared in Example 15.

[0020] Figure 4 This is a TEM image of the Mo,Ni-N-MCS prepared in Example 15.

[0021] Figure 5 The image shows the HAADF diagram of the Mo,Ni-N-MCS prepared in Example 15.

[0022] Figure 6 EDS Mapping (elemental distribution map) of Mo,Ni-N-MCS prepared in Example 15. Detailed Implementation

[0023] The present invention will be further described in detail below with reference to embodiments. These embodiments are merely illustrative of the technical features, objectives, and effects of the present invention, and are not intended to limit the scope of the invention. Example 1

[0024] A pharmaceutical composition for treating type 2 diabetic peripheral neuropathy, comprising 20g of Agrimonia pilosa, 12g of Oligospermum jasminoides, 12g of Spatholobus suberectus, and 5g of Hirudo medicinalis. Example 2

[0025] A pharmaceutical composition for treating type 2 diabetic peripheral neuropathy, comprising 10g of Agrimonia pilosa, 5g of Oligospermum jasminoides, 5g of Spatholobus suberectus, and 2g of Hirudo medicinalis. Example 3

[0026] A pharmaceutical composition for treating type 2 diabetic peripheral neuropathy, comprising 40g of Agrimonia pilosa, 20g of Oligospermum jasminoides, 20g of Spatholobus suberectus, and 9g of Hirudo medicinalis. Example 4

[0027] A pharmaceutical composition for treating type 2 diabetic peripheral neuropathy, comprising 10g of Agrimonia pilosa, 10g of Oligospermum jasminoides, 12g of Spatholobus suberectus, and 6g of Hirudo medicinalis. Example 5

[0028] A pharmaceutical composition for treating type 2 diabetic peripheral neuropathy, comprising 15g of Agrimonia pilosa, 12g of Oligospermum jasminoides, 15g of Spatholobus suberectus, and 8g of Hirudo medicinalis. Example 6

[0029] A pharmaceutical composition for treating type 2 diabetic peripheral neuropathy, comprising 20g of Agrimonia pilosa, 15g of Oligospermum jasminoides, 20g of Spatholobus suberectus, and 4g of Hirudo medicinalis. Example 7

[0030] A pharmaceutical composition for treating type 2 diabetic peripheral neuropathy, comprising 35g of Agrimonia pilosa, 20g of Oligospermum jasminoides, 5g of Spatholobus suberectus, and 2g of Hirudo medicinalis. Example 8

[0031] A pharmaceutical composition for treating type 2 diabetic peripheral neuropathy, comprising 40g of Agrimonia pilosa, 5g of Oligospermum jasminoides, 10g of Spatholobus suberectus, and 3g of Hirudo medicinalis. Example 9

[0032] A granule formulation of a pharmaceutical composition for treating type 2 diabetic peripheral neuropathy is prepared by weighing the medicinal materials according to any one of Examples 1-8, and following these steps: Soaking all the medicinal materials in 10 times the amount of water for 30 minutes, then refluxing for extraction, extracting for 1-2 hours each time, for 2-3 extractions; combining the filtrates, and concentrating under reduced pressure at 50-80℃ and 0.04-0.10 MPa to a thick paste with a relative density of 1.25-1.30 (60℃); adding 0.5 times the amount of water to the thick paste... Dextrin is mixed, dried (using at least one of freeze-drying, spray drying, vacuum drying, and pulsed vacuum drying), pulverized, and 0.5wt%-1.0wt% of sucralose is added. The total amount of dextrin is adjusted to twice the weight of the dry extract. Granulation is prepared using 60%-80% ethanol as a wetting agent. The granules are dried (using at least one of freeze-drying, spray drying, vacuum drying, and pulsed vacuum drying), granulated (using at least one of dry granulation, fluidized bed granulation, and wet granulation), and packaged to obtain the final product. Example 10

[0033] A pill formulation of a pharmaceutical composition for treating type 2 diabetic peripheral neuropathy is prepared by weighing the medicinal materials according to any one of Examples 1-8 and following these steps: soaking all the medicinal materials in 10 times the amount of water for 30 minutes, reflux extraction for 1-2 hours each time, and extracting 2-3 times; combining the filtrates, concentrating under reduced pressure at 50-80℃ and 0.04-0.10MPa to a thick paste with a relative density of 1.20-1.25 (60℃), drying, pulverizing, adding an appropriate amount of refined honey or water, mixing well, making a soft mass, forming pills using a plasticizing method, drying, and making pills. After passing inspection, the pills are packaged and sterilized to obtain the final product. Example 11

[0034] A pill formulation of a pharmaceutical composition for treating type 2 diabetic peripheral neuropathy is prepared by weighing the medicinal materials according to any one of Examples 1-8 and following these steps: Pulverize three medicinal materials—Potentilla anserina, Osmanthus fragrans, and Spatholobus suberectus—and pass them through a 120-mesh sieve; pulverize leeches separately and pass them through a 120-mesh sieve; mix the above-mentioned medicinal powders, add an appropriate amount of refined honey or water, form a soft mass, shape into pills using a molding method, dry, and prepare the pills. After passing inspection, package and sterilize to obtain the final product. Example 12

[0035] A concentrated pill formulation of a pharmaceutical composition for treating type 2 diabetic peripheral neuropathy is prepared by weighing the medicinal materials according to any one of Examples 1-8 and following these steps: soaking all the medicinal materials in 10 times the amount of water for 30 minutes, reflux extraction for 1-2 hours each time, and extracting 2-3 times; combining the filtrates, concentrating under reduced pressure at 50-80℃ and 0.04-0.10MPa to a thick paste with a relative density of 1.20-1.25 (60℃), vacuum drying at 40-60℃, pulverizing, adding excipients (starch or microcrystalline cellulose), mixing well, making concentrated pills by pan-coating, drying, and after passing inspection, dispensing and sterilizing to obtain the final product. Example 13

[0036] A concentrated pill formulation of a pharmaceutical composition for treating type 2 diabetic peripheral neuropathy is prepared by weighing the medicinal materials according to any one of Examples 1-8 and following these steps: Soak all the medicinal materials in 10 times the amount of water for 30 minutes, reflux extraction, 1-2 hours each time, for 2-3 extractions; combine the filtrates, concentrate under reduced pressure at 50-80℃ and 0.04-0.10 MPa to a thick paste with a relative density of 1.05-1.15 (60℃), add pharmaceutical ethanol to achieve an alcohol content of 50%-70%, let stand for 12-24 hours, filter, concentrate the filtrate under reduced pressure at 50-80℃ and 0.04-0.10 MPa to a thick paste with a relative density of 1.20-1.30 (60℃), vacuum dry at 40-60℃, pulverize, add excipients (starch or microcrystalline cellulose), mix well, prepare concentrated pills using a pan-coating method, dry, and after passing inspection, package and sterilize to obtain the final product. Example 14

[0037] Sustained-release microcapsules of a pharmaceutical composition for treating type 2 diabetic peripheral neuropathy are prepared by weighing medicinal materials according to any one of Examples 1-8, and following these steps: Soak all medicinal materials in 10 times their volume of water for 30 minutes, then reflux for extraction, repeating the extraction 2-3 times for 1-2 hours each time; combine the filtrates, concentrate under reduced pressure at 50-80℃ and 0.04-0.10 MPa to a thick paste with a relative density of 1.05-1.15 (60℃), add pharmaceutical ethanol to achieve an alcohol content of 50%-70%, let stand for 12-24 hours, filter, and concentrate the filtrate under reduced pressure at 50-80℃ and 0.04-0.10 MPa to a relative density of... A thick paste with a viscosity of 1.20-1.30 (60℃) is vacuum dried at 40-60℃, pulverized, mixed with microcrystalline cellulose, and prepared into microspheres using an extrusion-spheronization method with 60%-70% ethanol as a wetting agent. The extrusion speed is 45-60 r / min, the spheronization speed is 1500-2000 r / min, and the spheronization time is 5-10 min. The microspheres are then dried under reduced pressure at 60℃ for 4-6 h to form microspheres. These microspheres are then coated with 15% Sulis E-7-19050 aqueous dispersion at 40-60℃ with a spray pressure of 0.4-0.5 MPa and a spray rate of 1.0-1.5 mL / min for 1-2 h. After passing inspection, the microspheres are packaged and sterilized to obtain the final product. Example 15

[0038] A formulation of a pharmaceutical composition for treating type 2 diabetic peripheral neuropathy, loaded onto a mesoporous material, is prepared by the following steps: 1. Preparation of drugs for loading Take any of the pharmaceutical compositions described in Examples 1-8 and prepare them into concentrated pills according to the method described in Example 12 or Example 13. Name them "Yuqing Concentrated Pills" (Yuqing Concentrated Pills are concentrated pills prepared from the pharmaceutical compositions described in this invention) for later use.

[0039] 2. Preparation of supported mesoporous materials Mo, Ni-N-MCS This embodiment provides a method for preparing a mesoporous material for loading pharmaceutical active ingredients. The specific steps are as follows: 15.17 mL of a 28% (w / w) aqueous solution of hexadecyltrimethylammonium bromide is rapidly added to an aqueous solution containing 40 mL of ethanol, 95 mL of deionized water, and 0.5 mL of NH3·H2O. After vigorous stirring for 0.5 h, 1 g of resorcinol is added to the above solution, and the mixture is stirred at room temperature for 0.5 h. Then, 3.6 mL of tetraethyl orthosilicate (TEOS) and 1.4 mL of formaldehyde are added to the solution. After reacting at room temperature for 24 h, the resulting pink product is centrifuged, washed, and dried in a vacuum drying oven at 60 °C for 12 h. The dried solid is then placed in an atmosphere furnace under N2 protection and carbonized at 350 °C, 500 °C, and 800 °C for 2 h, 2 h, and 5 h, respectively, at a heating rate of 5 °C / min. After cooling to room temperature, the obtained product was etched with 8% HF and stirred for 24 hours, centrifuged (this step was repeated twice), washed repeatedly with deionized water by centrifugation 5 to 7 times, and dried in a vacuum drying oven at 60°C for 12 hours. The obtained product was N-doped mesoporous carbon material (N-MCS).

[0040] Take 20 mg of the N-MCS prepared above and disperse it in 4.0 mL of deionized water (double-distilled water), add 10 mg of dopamine hydrochloride and 20 mg of (NH4)6Mo7O 24 Mix thoroughly with 4H2O and sonicate for 10 minutes.

[0041] 3.6 mg NiCl2 and 5.56 mg 1,10-phenanthroline (CAS No.: 66-71-7) were dissolved in 28 mL of anhydrous ethanol. The solution was added, and the mixture was sonicated for 1 h, stirred at 60 °C for 3 h, washed by centrifugation with anhydrous ethanol, and dried in a vacuum drying oven at 60 °C for 12 h. The product was placed in an atmosphere furnace under N2 protection and carbonized at 350 °C, 500 °C, and 900 °C for 2 h, 2 h, and 4 h, respectively, at a heating rate of 5 °C / min. The obtained product was etched with 1.0 M HCl by stirring for 24 h, washed by centrifugation with anhydrous ethanol, and then vacuum dried for 12 h to obtain molybdenum, nickel, and nitrogen-doped mesoporous carbon material (Mo, Ni-N-MCS).

[0042] 3. Loading of drug extracts with Mo,Ni-N-MCS A drug-loaded sample supported on Mo,Ni-N-MCS was prepared using a solvent evaporation method. The specific procedures were as follows: 100 mg of concentrated molybdenum granules was weighed and dissolved in 10 mL of distilled water. The solution was then sonicated in a water bath to ensure complete dissolution, yielding an aqueous solution of concentrated molybdenum granules. Next, 100 mg of Mo,Ni-N-MCS was weighed and added to the above aqueous solution of concentrated molybdenum granules (the weight ratio of Mo,Ni-N-MCS mesoporous material to concentrated molybdenum granules can be 1:1 to 1:2; in this example, a 1:1 ratio was used). The resulting mixture was placed in a sealed container and stirred for 12 h. This process was carried out at room temperature in the dark. Finally, the sample was centrifuged at 8000 r / min for 10 min, washed three times with distilled water, and the supernatant was discarded. The collected solid sample was then frozen (at -40°C) and dried to remove residual solvent, yielding the concentrated molybdenum granules / Mo,Ni-N-MCS formulation.

[0043] 4. Morphology and structural characterization of Mo, Ni-N-MCS 4.1 SEM morphology characterization The microstructure of N-MCS and Mo,Ni-N-MCS was characterized by field emission scanning electron microscopy, and the results are as follows: Figure 1 and with Figure 2 As shown. Figure 1 The image shows a SEM image of N-MCS. As can be seen, the N-MCS particles are uniform in size, well-dispersed, and show almost no aggregation. All N-MCS particles exhibit a highly ordered state, with a particle size of approximately 339 nm. The surface of the microspheres has abundant and uniformly distributed micropores. These abundant mesopores facilitate drug penetration and metal growth, while the internal hollow structure provides ample active sites for the metal. Figure 2 The SEM image of Mo,Ni-N-MCS shows that the Mo,Ni-N-MCS is basically spherical with some damage. The diameter of the nanospheres is about 360 nm. Due to the doping of Mo and Ni, the particle size of the N-doped mesoporous carbon spheres increases. The N element in the mesoporous carbon structure provides active sites for the doping of Mo and Ni. The doping of Mo and Ni forms a bimetallic synergistic effect, which is beneficial to the loading of the prepared Mo,Ni-N-MCS onto the concentrated pellets.

[0044] 4.2 TEM and EDS Mapping Characterization The morphology and structure of N-MCS and Mo,Ni-N-MCS were further characterized by TEM, and the results are as follows: Figure 3 As shown. Figure 3 The image shows a TEM image of N-MCS. As can be seen from the image, the mesoporous carbon has a regular and full shape, uniform size, and a clear porous structure on the surface, indicating uniform dispersion.

[0045] Figure 4The TEM image of Mo,Ni-N-MCS shows that Ni and Mo are uniformly doped in the pores and surface of N-doped mesoporous carbon without obvious agglomeration. Figure 5 The image shows the HAADF plot of Mo,Ni-N-MCS. As can be seen from the figure, Ni and Mo are uniformly doped in the N-doped mesoporous carbon voids and on the surface. Figure 6 The EDS mapping diagram (elemental distribution diagram) of Mo,Ni-N-MCS clearly shows that C, N, O, Ni, and Mo elements are uniformly distributed on the spherical mesoporous carbon without obvious agglomeration.

[0046] 5. Determination of drug loading Berberine and astragaloside A are the characteristic active ingredients of the pharmaceutical composition described in this invention, and their content directly reflects the loading effect of the active pharmaceutical ingredient. 100 mg of Yuqing concentrated pills / Mo,Ni-N-MCS was accurately weighed into a glass container, 25 mL of anhydrous ethanol was added, and the mixture was sonicated for 15 min and stirred at room temperature for 12 h to ensure complete release of the main active ingredients from the Mo,Ni-N-MCS. An appropriate amount of the mixed solution was filtered through a 0.45 μm microporous membrane, and 2 mL of the filtrate was accurately measured and placed in a 10 mL volumetric flask, then diluted to volume with anhydrous ethanol. The contents of berberine and astragaloside A were then measured using high-performance liquid chromatography (HPLC). The determination was repeated 3 times, and the drug loading was calculated as follows: Drug loading = W0 / W × 100%, where W0 is the mass of berberine or astragaloside A contained in Yuqing Concentrated Pills / Mo,Ni-N-MCS, and W is the mass of Yuqing Concentrated Pills / Mo,Ni-N-MCS. The results are shown in Table 1.

[0047]

[0048] The average loading of berberine was measured to be 28.3%, and the average loading of astragaloside A was 25.5%, indicating that Mo,Ni-N-MCS has a high loading capacity for the active ingredients in the pharmaceutical composition of the present invention.

[0049] Application Example 1 - Study on the mechanism of the pharmaceutical composition of the present invention loaded with mesoporous material for treating type 2 diabetic peripheral neuropathy 1. Materials and Methods 1.1 Test Drug The pharmaceutical composition for treating type 2 diabetic peripheral neuropathy described in Example 8 was first prepared into concentrated pills according to the method described in Example 12, and then prepared into a formulation loaded with Mo,Ni-N-MCS mesoporous material (Yuqing concentrated pills / Mo,Ni-N-MCS formulation) according to the method described in Example 15. Before use, it was prepared into a suspension with sterile physiological saline and used immediately.

[0050] 1.2 Laboratory Animals Eighteen specific pathogen-free (SPF) adult healthy male C57BL / 6J rats, 6-8 weeks old, weighing 18-22 g, were purchased from Cyagen Biosciences Inc. The experimental animal production license number is SCXK (Ji) 2021-003.

[0051] 1.3 Main instruments and reagents

[0052]

[0053] 1.4 Experimental methods (1) Animal grouping: Normal control group, model group, and experimental group, with 6 animals in each group.

[0054] (2) Animal modeling: The normal control group was fed with standard feed, and the other two groups were given 60% high-fat feed. At the 12th week, except for the normal control group which was intraperitoneally injected with citrate buffer (pH 4.3), the other two groups were continuously injected with STZ (streptozotocin) (50 mg / kg, freshly prepared on ice in the dark before use) for 3 days. After the stable period, fasting blood glucose (FBG) was measured, and individuals with FBG > 11.1 mmol / L were selected for subsequent experiments.

[0055] (3) Administration method: From the 14th to the 22nd week, the normal control group and the model group were intragastrically administered with an equal volume of sterile normal saline every day, with an intragastric volume of 10 mL / kg; the experimental group was intragastrically administered with a suspension of Yuqing concentrated pills / Mo,Ni-N-MCS preparation. Calculated based on the raw drug content of the original drug composition, the administration dose was 18.2 g / kg, and the intragastric volume was 10 mL / kg.

[0056] Throughout the experiment, the body weight and fasting blood glucose of the rats were monitored weekly (blood was collected from the tail vein after 8 hours of fasting for detection).

[0057] 1.5 Index detection (1) Body weight monitoring: Before grouped feeding, at 12 weeks of grouped feeding, 1 week after modeling, and at the 4th and 8th weeks of treatment, the body weights of the rats in each group were weighed and recorded using an electronic balance.

[0058] (2) Blood glucose measurement and OGTT test: Blood glucose was measured using the tail vein blood collection method. The rat tail vein was disinfected with iodine-soaked cotton balls, and a disposable needle was used to puncture the capillary in the tail. The first drop of blood was discarded, and the test strip was placed on the bleeding site of the rat tail vein until the blood permeated the test area. The blood glucose meter read and recorded the data. Rats with successful modeling were fasted for 12 hours and then subjected to an oral glucose tolerance test (OGTT). 1 g / kg of anhydrous glucose was administered by gavage, and fasting blood glucose and blood glucose at 0.5, 1, 1.5, and 2 hours after the glucose load were measured. The area under the curve (AUC) of the OGTT test was calculated using the formula: AUC OGTT = 1 / 4 × 0h blood glucose + 1 / 2 × 0.5h blood glucose + 3 / 4 × 1h blood glucose + 1 / 2 × 2h blood glucose.

[0059] (3) Calculation of serum FINS level and HOMA-IR index: At the end of the experiment, fasting arterial blood was collected from rats, and serum was collected by centrifugation. Serum fasting insulin (FINS) level was detected by enzyme-linked immunosorbent assay (ELISA). The homeostatic model insulin resistance index was calculated based on fasting blood glucose (FBG) and FINS, HOMA-IR = FBG × FINS / 22.5.

[0060] (4) Blood lipid level measurement: Rat serum was collected and the levels of triglycerides (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C) were detected using a fully automated biochemical analyzer or corresponding reagent kit.

[0061] (5) Detection of serum inflammatory factors: The levels of pro-inflammatory factors (IL-1β, IL-6, TNF-α) and anti-inflammatory factors (IL-10) in rat serum were detected by ELISA. The operation steps were strictly performed in accordance with the kit instructions.

[0062] (6) Evaluation of vascular function of sciatic nerve tissue: The blood flow of the sciatic nerve in rats was detected by laser Doppler flowmeter and the percentage change in blood flow was calculated; the vascular density of the sciatic nerve cross section was observed and counted under a microscope by histopathological section or immunohistochemical staining.

[0063] (7) Measurement of nerve conduction velocity: Before the end of the experiment, the rats were anesthetized and fixed to expose the sciatic nerve. The motor nerve conduction velocity (MNCV) and sensory nerve conduction velocity (SNCV) of the rat sciatic nerve were measured using a biological signal acquisition and processing system.

[0064] (8) Detection of biochemical indicators of sciatic nerve: Sciatic nerve tissue from rats was dissected and tissue homogenate was prepared. Malondialdehyde (MDA) content was detected using the thiobarbituric acid method; superoxide dismutase (SOD) and glutathione peroxidase (GSH-PX) activities, as well as sodium-potassium pump (Na+) activity, were detected using reagent kits. +-K + The activity of ATPase was used to assess the level of oxidative stress and metabolic function in nerve tissue.

[0065] 1.6 Statistical Methods Experimental data were analyzed using GraphPad Prism 9.0, and the results were represented as follows: t Test the difference between the two groups, when P <0.05 indicates a statistically significant difference.

[0066] 2. Experimental Results 2.1 Effects on body weight and blood glucose in a rat model of type 2 diabetic peripheral neuropathy Before being grouped and fed, there was no significant difference in the body weight of the rats in each group. P >0.05); After 12 weeks of group feeding, the body weight of rats in the model group and experimental group was significantly higher than that in the normal control group ( P <0.01). One week after modeling, the body weight of rats in both the model group and the experimental group decreased to varying degrees compared with that at 12 weeks of group feeding; after the 4th and 8th weeks of treatment, the body weight of rats in the experimental group was significantly lower than that in the model group ( P <0.01), the difference was statistically significant (see Table 4). Meanwhile, it can be seen that after modeling, the fasting blood glucose level of rats in the model group was significantly higher than that in the normal control group ( P <0.01), and remained in a hyperglycemic state until the end of the experiment. After 4 weeks of treatment, the blood glucose levels of the experimental group rats were significantly lower than those of the model group ( P <0.01), after 8 weeks of treatment, the blood glucose level of the experimental group rats further decreased, significantly lower than that of the model group ( P <0.01 (see Table 5).

[0067]

[0068]

[0069] 2.2 Effect on the area under the OGTT curve Compared with the normal control group, the area under the OGTT curve in the model group rats was significantly increased ( P <0.01). Compared with the model group, the area under the OGTT curve in the experimental group rats was significantly reduced ( P <0.01 (see Table 6).

[0070]

[0071] 2.3 Effects on FINS levels and HOMA-IR index Compared with the normal control group, the levels of FINS and HOMA-IR in the model group rats were significantly increased (P <0.05, P <0.01). Compared with the model group, the levels of FINS and HOMA-IR in the experimental group rats were significantly decreased ( P <0.01 (see Table 7).

[0072]

[0073] 2.4 Effects on lipid metabolism in a rat model of type 2 diabetic peripheral neuropathy Compared with the normal control group, the levels of TG, TC and LDL in the model group rats were significantly increased. P <0.01), while HDL levels were significantly reduced ( P <0.01). Compared with the model group, the experimental group significantly reduced TG, TC and LDL levels ( P <0.01), while HDL levels have rebounded (see Table 8).

[0074]

[0075] 2.5 Effects on chronic low-grade inflammation in a rat model of type 2 diabetic peripheral neuropathy Compared with the normal control group, the levels of pro-inflammatory factors IL-1β, IL-6 and TNF-α in the model group rats were significantly increased. P <0.01), while the level of the anti-inflammatory factor IL-10 was significantly reduced ( P <0.01. Compared with the model group, the levels of IL-1β, IL-6 and TNF-α in the experimental group rats were significantly reduced ( P <0.05, P <0.01), while IL-10 levels increased significantly ( P <0.01 (see Table 9).

[0076]

[0077] 2.6 Effects on sciatic nerve tissue vascular dysfunction Compared with the normal control group, the percentage change in blood flow and cross-sectional vascular density in the model group rats were significantly reduced. P <0.01). Compared with the model group, the percentage change in blood flow and cross-sectional vascular density of rats in the experimental group were significantly increased ( P <0.05, P <0.01 (see Table 10).

[0078]

[0079] 2.7 Effects on nerve motor and sensory conduction velocities Compared with the normal control group, the model group showed a significant decrease in both motor and sensory conduction velocities. P <0.01. Compared with the model group, the experimental group rats showed significantly increased motor and sensory conduction velocities ( ). P <0.01 (see Table 11).

[0080]

[0081] 2.8 MDA and Na content in the sciatic nerve + -K + The role of ATPase, SOD, and GSH-PX activities Compared with the normal control group, the MDA content in the sciatic nerve of the model group was significantly increased, and Na... + -K + ATPase, SOD, and GSH-PX activities decreased significantly (all) P <0.01); Compared with the model group, the MDA content in the sciatic nerve of rats in the experimental group was significantly reduced, and Na + -K + ATPase, SOD, and GSH-PX activities were significantly increased (all) P <0.01 (see Table 12).

[0082]

[0083] 3. Conclusion The concentrated Yuqing pill / Mo,Ni-N-MCS formulation used in this embodiment can effectively improve the symptoms and functional indicators of peripheral neuropathy in a type 2 diabetic rat model by comprehensively regulating multiple pathological processes, including blood glucose, blood lipids, inflammation, oxidative stress, and nerve microcirculation. Its mechanism of action may be related to improving insulin resistance, alleviating metabolic disorders and inflammation, enhancing antioxidant defense, and promoting nerve blood supply and energy metabolism. Compared to conventional formulations, this formulation, while achieving the same efficacy, is expected to provide a more durable chemical regulatory environment and a favorable physical microenvironment for nerve repair through its sustained-release properties and physical support, thus offering a more promising treatment strategy for reversing or delaying type 2 diabetic peripheral neuropathy.

[0084] Application Example 2 – Clinical Study of the Pharmaceutical Composition of the Invention for the Treatment of Type 2 Diabetic Peripheral Neuropathy 1. Materials and Methods 1.1 General Information Patients with type 2 diabetic peripheral neuropathy treated at the Department of Endocrinology, Yunnan Provincial Hospital of Traditional Chinese Medicine from June 2022 to April 2024 were included in this study. They were randomly divided into a control group and an observation group, with 100 patients in each group. The actual number of patients enrolled was 102 in the observation group and 98 in the control group. In the observation group, there were 86 males and 16 females, with an average age of 47.00 (39.00, 55.00) years, a duration of T2DM with obesity of 9.00 (6.00, 11.00) years, and obesity grades: mild (64 cases), moderate (23 cases), and severe (15 cases). In the control group, there were 84 males and 14 females, with an average age of 48.50 (41.75, 54.00) years, a duration of T2DM with obesity of 9.00 (6.00, 12.00) years, and obesity grades: mild (64 cases), moderate (21 cases), and severe (13 cases). There were no statistically significant differences between the two groups of patients in terms of age, duration of obesity-related type 2 diabetes mellitus, sex, and obesity grade. P >0.05), making them comparable (see Tables 13 and 14).

[0085]

[0086]

[0087] 1.2 Diagnostic criteria (1) Western medicine diagnostic criteria Diagnostic criteria were developed in accordance with the "Guidelines for the Prevention and Treatment of Type 2 Diabetes in China (2020 Edition)" and domestic and international consensus on the diagnosis and treatment of diabetic peripheral neuropathy. ① Has a clear history of type 2 diabetes; ② Symptoms and / or signs of peripheral neuropathy that appear at or after the diagnosis of diabetes, such as symmetrical numbness, pain, and paresthesia in the distal extremities; ③ Abnormalities in neurological function tests: Any one of the following five tests is sufficient for diagnosis: abnormal ankle reflex, abnormal pinprick pain sensation, abnormal temperature sensation, abnormal vibration sensation, and abnormal pressure sensation of 10g nylon filament. ④ Exclude other causes of peripheral neuropathy (such as cervical and lumbar spine lesions, cerebral infarction, Guillain-Barré syndrome, severe arteriovenous vascular disease, drug-induced, etc.).

[0088] (2) Traditional Chinese Medicine diagnostic criteria Based on the "Guidelines for Clinical Diagnosis and Treatment of Diabetic Peripheral Neuropathy in Traditional Chinese Medicine (2016 Edition)" and the "Guidelines for Diagnosis and Treatment of Diabetic Peripheral Neuropathy Combining Disease and Syndrome (2021 Edition)," the diagnostic criteria for Qi and Yin deficiency with damp-heat stagnation syndrome are proposed as follows: Main symptoms: ① numbness in the limbs, like ants crawling; ② pain in the limbs, tingling or burning pain, especially at night; ③ decreased sensation, cold or burning sensation in the limbs.

[0089] Secondary symptoms: ① Fatigue, lack of energy, and reluctance to speak; ② Dry mouth and throat, and hot flashes in the palms, soles, and chest; ③ Dull complexion and dark lips and nails.

[0090] Tongue and pulse: The tongue is dark red or has petechiae or ecchymosis, and the coating is thin and white or has little coating; the pulse is thready and hesitant or weak.

[0091] Diagnostic criteria: If at least two primary symptoms and at least two secondary symptoms are present, combined with tongue and pulse manifestations, the syndrome can be diagnosed as Qi and Yin deficiency with damp-heat stagnation.

[0092] 1.3 Inclusion Criteria (1) Meets the above-mentioned Western medicine diagnostic criteria for type 2 diabetic peripheral neuropathy; (2) Meets the above-mentioned TCM diagnostic criteria for Qi and Yin deficiency and damp-heat stagnation syndrome; (3) Age between 18 and 75 years old (inclusive), gender not limited; (4) Relatively stable blood glucose control at the time of enrollment: glycated hemoglobin (HbA1c) ≤ 8.5%, or fasting plasma glucose (FPG) ≤ 10.0 mmol / L; (5) Toronto Clinical Scoring System (TCSS) score ≥6, and nerve conduction velocity (NCV) test indicates peripheral neuropathy (at least one nerve conduction velocity is below the lower limit of normal). (6) Have not used neurotrophic drugs (such as mecobalamin, thioctic acid, epalrestat, etc.) or drugs that improve microcirculation (such as alprostadil, kallikrein, etc.) in the past month. (7) Has not participated in any other drug clinical trials in the past 3 months; (8) Voluntarily participate in this study and sign the informed consent form.

[0093] 1.4 Exclusion Criteria (1) Individuals with extremely unstable blood glucose control: Fasting plasma glucose (FPG) > 13.0 mmol / L, or glycated hemoglobin (HbA1c) > 10.0%; (2) Those who have acute complications such as diabetic ketoacidosis or hyperosmolar hyperglycemia, or who have experienced severe infection, trauma or other stress within the past month; (3) Other causes of peripheral neuropathy (such as cervical and lumbar spine lesions, cerebral infarction, Guillain-Barré syndrome, uremia, drug-induced, alcoholic, hereditary, etc.); (4) Those with severe cardiovascular and cerebrovascular diseases (such as unstable angina pectoris, myocardial infarction, severe arrhythmia, stroke), liver and kidney dysfunction (ALT or AST > 1.5 times the upper limit of normal, Cr > the upper limit of normal), hematopoietic system diseases or malignant tumors; (5) Has a history of mental illness or cognitive impairment and is unable to cooperate with the researcher; (6) Pregnant or breastfeeding women, or those who plan to have children within the next 3 months; (7) Individuals with known allergies to the drugs studied (Potentilla chinensis, Oligospermum jasminoides, Spatholobus suberectus, Hirudo medicinalis) or their excipients; (8) Those who are currently participating in other clinical trials, or who have participated in other clinical trials within the past 3 months.

[0094] 1.5 Treatment Methods All enrolled patients underwent a 2-week introductory period before enrollment, during which they maintained their original glucose-lowering regimen (metformin monotherapy) and received lifestyle guidance (including dietary control and appropriate exercise). After the introductory period, patients meeting the inclusion criteria were randomly assigned to the control group and the observation group in a 1:1 ratio.

[0095] Control group (positive control): In addition to the existing oral metformin hydrochloride extended-release tablets, patients received oral mecobalamin tablets (Ruiyang Pharmaceutical Co., Ltd., specification: 0.5mg, batch number: 20210121). Dosage and administration: 0.5mg each time, 3 times a day, for a course of 3 months.

[0096] Observation group: In addition to maintaining the original oral treatment with metformin hydrochloride extended-release tablets (dosage and administration as in the control group), the drug composition described in Example 7 of this invention (prepared as granules according to the method described in Example 9) was added. Administration: 3 times daily, warm, for a course of 3 months.

[0097] 1.6 Observation Indicators and Methods (1) Evaluation criteria for TCM syndrome differentiation and treatment efficacy Evaluation was conducted after 8 weeks of treatment. Efficacy was assessed based on the "Guiding Principles for Clinical Research of New Traditional Chinese Medicines" and the improvement in neuroelectrophysiological parameters: ① Markedly effective: Total TCM syndrome score decreased by ≥70%, and the conduction velocity of major limb nerves (median nerve, common peroneal nerve) returned to normal or increased by >5 m / s compared to before treatment; ② Effective: Total TCM syndrome score decreased by ≥30% but <70%, and the conduction velocity of major limb nerves increased by 2~5 m / s; ③ Ineffective: The above criteria were not met. Total effective rate = (Number of markedly effective cases + Number of effective cases) / Total number of cases × 100%.

[0098] (2) Toronto Clinical Scoring System (TCSS) assessment Before treatment and 8 weeks after treatment, two trained physicians independently assessed patients using the TCSS scale. The scale consists of three parts: sensory function (ankle reflex, pinprick pain sensation, temperature sensation, light touch sensation, vibration sensation, and position sense, 0-1 points for each item), deep nerve reflexes (knee reflex and ankle reflex, 0-2 points for each item), and clinical symptoms (lower limb weakness, paresthesia, pain, numbness, and ataxia, 0-2 points for each item), with a total score of 0-19 points. Higher scores indicate more severe neurological impairment.

[0099] (3) Traditional Chinese Medicine syndrome score assessment Evaluations were conducted before treatment and 8 weeks after treatment, in accordance with the "Guiding Principles for Clinical Research of New Traditional Chinese Medicine Drugs (Trial Implementation)". The main symptoms included limb numbness, limb pain, fatigue, dry mouth and throat, hot flashes in the palms, soles, and chest, dull complexion, and dark lips and nails. Each symptom was scored according to severity: no symptoms (0 points); mild, occasional, not affecting daily life (2 points); moderate, frequent, slightly affecting daily life (4 points); severe, persistent, seriously affecting daily life (6 points). The total TCM syndrome score was the sum of the scores for each symptom.

[0100] (4) Measurement of weight, body mass index (BMI), and waist-to-hip ratio (WHR) The patient was fasting, barefoot, and wearing light clothing. Their height (m) and weight (kg) were measured using the same height and weight scale. BMI = weight / height 2 Use a soft measuring tape to measure waist circumference (the horizontal level of the midpoint of the line connecting the lower edge of the rib arch and the iliac crest) and hip circumference (the horizontal level of the most prominent part of the buttocks), accurate to 0.1cm. WHR = waist circumference / hip circumference.

[0101] (5) Measurement of blood glucose, insulin and lipid metabolism indicators All patients fasted for 8-12 hours before having blood drawn from their antecubital vein in the early morning.

[0102] Fasting plasma glucose (FPG) and 2-hour postprandial glucose (2hPG): measured using the glucose oxidase method and a fully automated biochemical analyzer.

[0103] Glycated hemoglobin (HbA1c): determined by high performance liquid chromatography (HPLC).

[0104] Fasting insulin (FINS): Measured using chemiluminescent immunoassay.

[0105] Insulin resistance index: HOMA-IR = [FPG (mmol / L) × FINS (μU / mL)] / 22.5.

[0106] The four lipid tests, including total cholesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C), were all measured using an enzymatic method and a fully automated biochemical analyzer.

[0107] (6) Measurement of blood rheological parameters Three mL of fasting antecubital venous blood was collected from the patient and anticoagulated with heparin. Whole blood viscosity at high shear rate (200 s⁻¹) was measured using an automated blood rheometer. -1 ), whole blood viscosity low shear (1s) -1The results, including plasma viscosity, were expressed in millipascals per second (mPa·s) and were strictly controlled at a constant temperature of 25°C.

[0108] (7) Measurement of nerve conduction velocity Electromyography / evoked potential testing was performed before treatment and 8 weeks after treatment at room temperature (22-25℃). Patients were placed in a supine position with their limbs relaxed.

[0109] Motor nerve conduction velocity (MNCV): Stimulating electrodes are placed proximally to the nerve (e.g., stimulating the common peroneal nerve at the popliteal fossa, or stimulating the median nerve at the elbow), and recording electrodes are placed distally to the corresponding muscles (e.g., extensor digitorum brevis, abductor hallucis brevis). The latency of the compound muscle action potential and the distance between the stimulation points are measured, and MNCV is calculated.

[0110] Sensory nerve conduction velocity (SNCV): Using the antegrade method, the stimulating electrode is placed at the finger (toe) tip, and the recording electrode is located at the proximal end of the nerve trunk (such as the wrist or ankle). The latency and distance of the sensory nerve action potential are measured, and the SNCV is calculated.

[0111] (8) Safety evaluation Record any adverse events that occur during treatment. Fasting venous blood samples were collected before treatment and 8 weeks after treatment to measure complete blood count, urinalysis, liver function (ALT, AST), and kidney function (BUN, Cr) to assess the safety of treatment.

[0112] 1.7 Statistical Methods Statistical analysis was performed on the data using SPSS 28.0. Normally distributed data were expressed as , non-normally distributed data as Median (P25, P75), and count data as . n (%) indicates the data. The Kolmogorov-Smirnov method was used to test whether the variables followed a normal distribution for continuous data. Paired samples were used for within-group comparisons. t Tests and between-group comparisons use two independent samples. t Tests were performed. For non-normally distributed data, the Wilcoxon test was used for within-group comparisons, and the Mann-Whitney U test was used for between-group comparisons. For continuous data, the nonparametric rank-sum test was used. P <0.05 indicates a statistically significant difference.

[0113] 2. Results 2.1 Comparison of TCM syndrome differentiation and treatment efficacy between the two groups of patients Compared with the control group, the total effective rate of the observation group was significantly higher than that of the control group, and the difference was statistically significant. P <0.05 (see Table 15).

[0114]

[0115] 2.2 Comparison of neurological function scores before and after treatment in the two groups of patients Compared with pre-treatment levels, both groups of patients showed significant decreases in sensory function, deep nerve reflexes, and clinical symptom scores, with statistically significant differences. P <0.01. Compared with the control group, the observation group showed significantly better improvement in sensory function, deep nerve reflexes, and clinical symptoms after treatment, with statistically significant differences. P <0.05 (see Table 16).

[0116]

[0117] 2.3 Comparison of TCM syndrome scores before and after treatment in the two groups of patients Compared with before treatment, after treatment, the scores of all symptoms (limb numbness, limb pain, fatigue, dry mouth and throat, hot flashes in the palms, soles, and chest), sallow complexion, and dark lips and nails) as well as the total score of TCM syndrome in the observation group were significantly reduced, and the differences were statistically significant. P <0.05); In the control group, the scores for limb numbness, limb pain, fatigue, and total TCM syndrome score were significantly lower than before treatment, and the differences were statistically significant. P <0.05), and the scores of other symptoms showed no significant change compared to before treatment ( P >0.05). In post-treatment intergroup comparisons, the scores for each symptom (limb numbness, limb pain, fatigue, dry mouth and throat, hot flashes in the palms, soles, and chest), sallow complexion, and dark lips and nails, as well as the total TCM syndrome score, were significantly lower in the observation group than in the control group; all differences were statistically significant. P <0.05 (see Table 17).

[0118]

[0119] 2.4 Comparison of body weight, BMI, and WHR between the two groups of patients before and after treatment Compared with pre-treatment levels, both groups of patients showed significant reductions in body weight, BMI, and WHR, with statistically significant differences. P <0.05). After treatment, the body weight and BMI of the patients in the observation group were significantly lower than those in the control group, and the difference was statistically significant. P <0.05 (see Table 18).

[0120]

[0121] 2.5 Comparison of blood glucose and insulin levels before and after treatment in the two groups of patients Compared with pre-treatment levels, FPG, 2hPG, HbA1c, FINS, and HOMA-IR were all significantly lower in both groups, with statistically significant differences. P<0.05). Compared with the control group, the observation group showed significantly lower FPG, 2hPG, HbA1c, FINS, and HOMA-IR after treatment, with statistically significant differences. P <0.05 (see Table 19).

[0122]

[0123] 2.6 Comparison of blood lipids before and after treatment in the two groups of patients Compared with pre-treatment levels, the levels of TC, TG, and LDL in the observation group were significantly lower, and the differences were statistically significant. P <0.05). Compared with the control group, the levels of TC, TG, and LDL in the observation group were significantly lower after treatment, and the differences were statistically significant. P <0.05 (see Table 20).

[0124]

[0125] 2.7 Comparison of hemorheological parameters between the two groups of patients before and after treatment Compared with pre-treatment levels, both groups of patients showed significant decreases in whole blood viscosity (high-shear, low-shear) and plasma viscosity, with statistically significant differences. P <0.01. Compared with the control group, the observation group showed significantly better improvement in both high and low shear viscosity of whole blood after treatment, and the difference was statistically significant. P <0.01 (see Table 21).

[0126]

[0127] 2.8 Comparison of nerve conduction velocities before and after treatment in the two groups of patients Compared with before treatment, the sensory and motor conduction velocities of the common peroneal nerve and median nerve were significantly improved in both groups of patients, and the differences were statistically significant. P <0.05. Compared with the control group, the observation group showed significantly better improvement in the peroneal nerve sensory conduction velocity, median nerve sensory conduction velocity, peroneal nerve motor conduction velocity, and median nerve motor conduction velocity, with statistically significant differences. P <0.05 (see Table 22).

[0128]

[0129] 2.9 Comparison of safety indicators between the two groups of patients No abnormalities were observed in blood routine tests, urine routine tests, liver function, or kidney function in either group of patients after treatment.

[0130] 3. Conclusion The pharmaceutical composition and its formulation described in this invention are used to treat type 2 diabetic peripheral neuropathy, demonstrating significant clinical efficacy. It effectively improves traditional Chinese medicine symptoms such as limb numbness, limb pain, fatigue, dry mouth and throat, hot flashes in the palms, soles, and chest, dull complexion, and dark lips and nails. Simultaneously, it significantly reduces body weight and BMI, regulates glucose and lipid metabolism disorders, and improves insulin resistance. Furthermore, by reducing whole blood and plasma viscosity, improving blood rheology and microcirculation, this drug promotes the repair and functional recovery of peripheral nerve structures, increases nerve conduction velocity, and significantly alleviates symptoms such as limb numbness and pain, ultimately achieving holistic intervention and effective treatment for type 2 diabetic peripheral neuropathy.

Claims

1. A pharmaceutical composition for treating type 2 diabetic peripheral neuropathy, characterized in that, By weight, it consists of the following components: 10-40 parts of Agrimonia pilosa, 5-20 parts of Gynostemma pentaphyllum, 5-20 parts of Spatholobus suberectus, and 2-9 parts of Hirudo medicinalis.

2. The pharmaceutical composition for treating type 2 diabetic peripheral neuropathy according to claim 1, characterized in that, By weight, it consists of the following components: 15-35 parts of Potentilla chinensis, 10-20 parts of Oligospermum jasminoides, 10-20 parts of Spatholobus suberectus, and 3-8 parts of Hirudo medicinalis.

3. The pharmaceutical composition for treating type 2 diabetic peripheral neuropathy according to claim 2, characterized in that, By weight, it consists of the following components: 20-35 parts of Agrimonia pilosa, 10-15 parts of Gynostemma pentaphyllum, 10-15 parts of Spatholobus suberectus, and 4-6 parts of Hirudo medicinalis.

4. A formulation of the pharmaceutical composition for treating type 2 diabetic peripheral neuropathy according to any one of claims 1 to 3, characterized in that, The formulation is prepared by adding pharmaceutically acceptable excipients to the pharmaceutical composition for treating type 2 diabetic peripheral neuropathy.

5. The formulation of the pharmaceutical composition for treating type 2 diabetic peripheral neuropathy according to claim 4, characterized in that, The formulation is a formulation in which a drug is loaded onto a mesoporous material.

6. The formulation of the pharmaceutical composition for treating type 2 diabetic peripheral neuropathy according to claim 5, characterized in that, The mesoporous material is a Mo, Ni, N co-doped mesoporous material.

7. The formulation of the pharmaceutical composition for treating type 2 diabetic peripheral neuropathy according to claim 6, characterized in that, The weight ratio of the Mo, Ni, and N co-doped mesoporous material to the loaded drug is 1:1 to 1:

2.

8. A method for preparing a formulation of the pharmaceutical composition for treating type 2 diabetic peripheral neuropathy according to claim 7, characterized in that, The process includes the following steps: (1) preparing the pharmaceutical composition into concentrated pills; (2) preparing Mo,Ni-N-MCS, a mesoporous material co-doped with Mo,Ni, and N; (3) dissolving the concentrated pills, adding Mo,Ni-N-MCS, stirring and mixing, centrifuging, washing with water, and freeze-drying to obtain the final product. The specific steps of step (2) are as follows: 15.17 mL of 28% hexadecyltrimethylammonium bromide aqueous solution is rapidly added to an aqueous solution containing 40 mL of ethanol, 95 mL of deionized water and 0.5 mL of NH3·H2O; after vigorous stirring for 0.5 h, 1 g of resorcinol is added, and the mixture is stirred at room temperature for 0.5 h, followed by the addition of 3.6 mL of tetraethyl orthosilicate and 1.4 mL of formaldehyde; after reacting at room temperature for 24 h, the resulting pink product is centrifuged, washed, and vacuum dried at 60 °C; the dried solid is carbonized under N2 protection at 350 °C, 500 °C and 800 °C for 2 h, 2 h and 5 h respectively, with a heating rate of 5 °C / min; after cooling to room temperature, the resulting product is etched with 8% HF and stirred for 24 h, centrifuged, repeated twice, washed repeatedly with deionized water, and vacuum dried at 60 °C. The resulting product is N-MCS; 20 mg of N-MCS is taken. N-MCS was dispersed in 4.0 mL of deionized water, and 10 mg of dopamine hydrochloride and 20 mg of (NH4)6Mo7O were added. 24 ·4H2O, mix thoroughly; dissolve 3.6 mg NiCl2 and 5.56 mg 1,10-phenanthroline in 28 mL of anhydrous ethanol, add to the above thoroughly mixed solution, sonicate for 1 h, stir at 60 °C for 3 h, wash with anhydrous ethanol by centrifugation, and dry under vacuum at 60 °C; place the product in an atmosphere furnace, under N2 protection, and carbonize at 350 °C, 500 °C, and 900 °C for 2 h, 2 h, and 4 h respectively, with a heating rate of 5 °C / min; etch the obtained product with 1.0 M HCl by stirring for 24 h, wash with anhydrous ethanol by centrifugation, and dry under vacuum to obtain Mo, Ni-N-MCS.

9. The use of the pharmaceutical composition for treating type 2 diabetic peripheral neuropathy according to any one of claims 1 to 3, characterized in that, The pharmaceutical composition for treating type 2 diabetic peripheral neuropathy is used in the preparation of a medicament for treating type 2 diabetic peripheral neuropathy.

10. The use of a formulation of the pharmaceutical composition of claim 4 for treating type 2 diabetic peripheral neuropathy, characterized in that, The formulation of the pharmaceutical composition for treating type 2 diabetic peripheral neuropathy is used in the preparation of a medicament for treating type 2 diabetic peripheral neuropathy.

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