Use of Shenqi Tongmai preparation in preparing medicine for preventing and treating diabetic neuropathy

Abstract

The present invention belongs to the field of traditional Chinese medicine, and specifically relates to the use of a Shenqi Tongmai preparation in the preparation of treatment for diabetic neuropathy. The diabetic neuropathy described is mainly lower limb vascular neuropathy. The Shenqi Tongmai preparation of the present invention contains various Chinese medicinal materials of plant medicine and animal medicine for synergistic enhancement. The Chinese medicinal composition described in pharmacology and clinical cases can reduce blood sugar levels, regulate nerve conduction velocity, improve peripheral neuropathy caused by painful diabetic neuropathy, especially diabetic foot, with rapid onset of treatment, high total effective rate, and high market value.

Description

Technical Field

[0001] The invention relates to use of a Shenqi Tongmai preparation in preparing a medicine for preventing and treating diabetic neuropathy, and belongs to the field of traditional Chinese medicines. Background Art

[0002] Diabetic peripheral neuropathy (DPN) is one of the common complications of diabetes. According to statistics, the morbidity of diabetes-induced DPN in patients worldwide can reach 24.4%-78.8%; among Chinese patients, about 80% of DPN is caused by diabetes, of which about 25% of patients clinically present with painful diabetic neuropathy. Because the disease invades the sensory nerves, typical diabetic painful neuropathy patients are characterized by spontaneous pain, hyperalgesia and abnormal pain, which can manifest as ant crawling, burning, acupuncture, stabbing, jumping, knife cutting, tearing, etc., accompanied by muscle weakness, directly involving peripheral nerves and central nerves, and obvious pain at night, which seriously affects the quality of daily life, restricts daily activities, and may cause depression in patients and affect social interaction.

[0003] The clinical manifestations of diabetic foot are neurological and vascular lesions, including sensory changes (stocking-like sensation, weakened light touch, proprioception, temperature sensation and pain perception), atrophy of intrinsic foot muscles; normal sweating of the skin, loss of temperature and blood regulation function: local ulcers, infection, osteomyelitis, Charcot arthropathy, etc.

[0004] Recent studies have found that the application of traditional Chinese medicine to diabetic patients can not only reduce the patient's blood sugar, but also improve the patient's clinical symptoms and prevent and treat their chronic complications; the effect of using traditional Chinese medicine to lower blood sugar is mostly not as good as Western medicine, but it has a significant effect on the treatment of vascular complications in diabetic patients. At the same time, traditional Chinese medicine can treat the AGEs-RAGE system in many ways, such as inhibiting the formation of AGEs, and inhibiting the expression of RAGE and its downstream related signal transduction.

[0005] In view of this, there is an urgent clinical need to provide a drug for treating diabetic painful neuropathy with small side effects, significant efficacy and low recurrence rate. Summary of the invention

[0006] In view of the shortcomings of the existing technology, a comprehensive study was conducted on Shenqi Tongmai Decoction, and the entire formula was innovated, from the original 5 herbs to 12 herbs. The present invention, with a scientific and rigorous attitude, confirmed through precise pharmacological experiments that the Shenqi Tongmai preparation has excellent therapeutic effects on the treatment of peripheral neuropathy, especially diabetic foot, with few side effects and low recurrence rate, and has great commercial value.

[0007] One of the purposes of the present invention is to provide a new type of Shenqi Tongmai preparation, which can be an oral preparation, preferably, the oral preparation is a decoction, a pill, a tablet, a mixture, a capsule, a granule, a powder or an ointment, more preferably a tablet or a pill, and most preferably a tablet.

[0008] Based on clinical experiments, decoctions, tablets, pills, etc., as long as they are prepared using the following technical solutions of the present invention, can achieve the same or similar therapeutic effects.

[0009] Specifically, the technical objectives of the present invention are achieved through the following technical solutions:

[0010] A Shenqi Tongmai preparation composed of raw astragalus, white atractylodes, cyathula, coix seed, angelica, salvia miltiorrhiza, millettia reticulata, honeysuckle, codonopsis pilosula, earthworm, scorpion and raw licorice is used in the preparation of a drug for preventing and treating diabetic neuropathy.

[0011] The components of the Chinese medicine composition are:

[0012]

[0013] Preferably, the components of the Shenqi Tongmai preparation are:

[0014]

[0015] The second object of the present invention is to provide the pharmaceutical use of the Shenqi Tongmai preparation. The neuropathy is painful diabetic peripheral neuropathy and lower limb vascular nerves.

[0016] Furthermore, the diabetic neuropathy is mainly lower limb vascular neuropathy. The Shenqi Tongmai preparation of the present invention contains various Chinese medicinal materials of plant medicine and animal medicine that synergistically enhance the effect. In pharmacology and clinical cases, the Chinese medicine composition of the present invention can lower blood sugar levels, regulate nerve conduction rate, and improve peripheral neuropathy caused by painful diabetic neuropathy, especially diabetic foot. The treatment has a fast onset and a high total effective rate, and has a high market value.

[0017] Furthermore, the diabetes described in the present invention is type 1 diabetes or type 2 diabetes.

[0018] Furthermore, the lower limb blood vessels and nerves described in the present invention are diabetic foot.

[0019] Furthermore, the diabetic foot described in the present invention refers to a disease state in which ulcers and gangrene occur in the feet of diabetic patients due to neuropathy that reduces the protective function of the lower limbs, and large blood vessel and microvascular lesions that cause insufficient arterial perfusion and microcirculatory disorders. It is a complication of diabetes.

[0020] The third object of the present invention is to provide a preparation of the above-mentioned Chinese medicine composition, wherein the Chinese medicine composition is prepared from the Chinese medicine components into a Chinese medicine oral preparation or an external preparation. Preferably, the Shenqi Tongmai preparation is an oral preparation. Preferably, the oral preparation is a decoction, pill, tablet, mixture, capsule, granule, powder or paste, more preferably a tablet.

[0021] Furthermore, the present invention also provides a method for preparing the Shenqi Tongmai preparation:

[0022] Take earthworms and scorpions, and crush them to a particle size of less than 100 μm;

[0023] Raw Astragalus, Atractylodes macrocephala, Cyathula officinalis, Coix seeds, Angelica sinensis and Codonopsis pilosula are added with 65-85% ethanol for extraction 1-2 times, each extraction takes 1.5-3.5 hours, filtered, ethanol is recovered from the filtrate, and the extract is concentrated under reduced pressure to a relative density of 1.05-1.10; the filtered residue is then extracted 1-2 times with 2-5 times water, filtered, and the water extract is reserved;

[0024] Extract volatile oil from honeysuckle, salvia miltiorrhiza, liquorice and millettia spatholobi with water, add 1 / 2 amount of earthworm and scorpion to the remaining liquid and residue, and boil for 1-2 times, each time for 1.5-3.5 hours, add ethanol to the filtrate and the water extract in step B to make the alcohol content reach 40-50%, stand at room temperature for 24-48 hours, recover ethanol from the filtrate, concentrate to an extract with a relative density of 1.15-1.20, and set aside;

[0025] The remaining 1 / 2 amount of earthworm and scorpion is mixed with the alcohol extract of step B and the water extract of step C, and a clinically acceptable dosage form is prepared directly or by adding a pharmaceutically acceptable excipient through conventional procedures.

[0026] Furthermore, the preparation method of the Shenqi Tongmai preparation of the present invention is:

[0027] Take earthworms and scorpions, and crush them to a particle size of less than 100 μm;

[0028] Raw Astragalus, Atractylodes macrocephala, Cyathula officinalis, Coix seeds, Angelica sinensis and Codonopsis pilosula are extracted twice with 75% ethanol, each time for 2.5 hours, filtered, the filtrate is used to recover ethanol, and the extract is concentrated under reduced pressure to a relative density of 1.08; the filtered residue is then extracted twice with 3 times water, filtered, and the water extract is reserved;

[0029] Extract volatile oil from honeysuckle, salvia miltiorrhiza, liquorice and millettia spatholobi with water, add 1 / 2 amount of earthworm and scorpion to the remaining liquid and residue, and boil twice, each time for 2.5 hours, add ethanol to the filtrate and the water extract in step B to make the alcohol content reach 45%, stand at room temperature for 48 hours, recover ethanol from the filtrate, concentrate to an extract with a relative density of 1.18, and set aside;

[0030] The remaining 1 / 2 amount of earthworm and scorpion is mixed with the alcohol extract of step B and the water extract of step C, and a clinically acceptable dosage form is prepared directly or by adding a pharmaceutically acceptable excipient through conventional procedures.

[0031] The invention has all the monarch, minister, assistant and envoy, proper compatibility and hits the mechanism. The monarch drugs in the prescription are: raw astragalus and atractylodes. These medicinal materials are the main ingredients that play a core role in the Tongmai prescription. Raw astragalus has the effects of invigorating qi and nourishing blood, replenishing qi and strengthening the exterior; atractylodes has the effects of strengthening the spleen and stopping diarrhea, and replenishing the spleen and stopping bleeding; the minister drugs are: cyathula, millettia reticulata, earthworm and scorpion. These medicinal materials play the role of auxiliary treatment and enhancing effect in the Tongmai prescription. Adjuvants: Coix seeds, Chinese angelica, salvia miltiorrhiza, honeysuckle, codonopsis, coix seeds have the effects of diuresis, swelling, clearing away heat and detoxification; Chinese angelica can promote blood circulation, regulate menstruation, nourish blood; salvia miltiorrhiza has the effects of promoting blood circulation, removing blood stasis, relaxing tendons and activating collaterals; Millettia reticulata can promote blood circulation, remove blood stasis, reduce swelling and relieve pain; honeysuckle has the effects of clearing heat and detoxification, detoxification and swelling; codonopsis has the effects of invigorating qi and strengthening spleen, and improving immunity; earthworm can promote blood circulation, dredge collaterals, dispel wind and dampness; scorpion has the effects of promoting blood circulation, removing blood stasis and relieving pain. Envoy: Licorice, in the prescription of dredge meridians, plays the role of harmonizing the medicinal properties, increasing the stability of the drug and promoting absorption.

[0032] Compared with the prior art, the present invention has the following advantages:

[0033] This study intends to give patients with diabetic ulcers of qi deficiency and blood stasis type oral Shenqi Tongmai Tablets, detect the changes in the downstream factors of AGEs-RAGE and the healing of wounds at different time points in patients, further verify the effectiveness and safety of Shenqi Tongmai Tablets in treating diabetic ulcer patients, and promote clinical application. It can be seen that the Chinese medicine composition of the present invention has a reasonable formula and has a significant therapeutic effect on neuropathic pain. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] Figure 1 The present invention shows the analysis of wound healing in Example 1 and Comparative Examples 1-3.

[0035] Figure 2 Shown are changes in wound surface area in Example 1 and Comparative Examples 1-3.

[0036] Figure 3 It shows the comparison of wound surface area of ​​Example 1 and Comparative Examples 1-3 after 3 weeks of treatment.

[0037] Figure 4 The figure shows the comparison of wound surface area of ​​Example 1 and Comparative Examples 1-3 after 4 weeks of treatment.

[0038] Figure 5 Shown are the changes in TNF-a in Example 1 and Comparative Examples 1-3.

[0039] Figure 6Shown are the changes in IL-6 in Example 1 and Comparative Examples 1-3.

[0040] Figure 7 It shows the changes of high-sensitivity C-reactive protein in Example 1 and Comparative Examples 1-3. DETAILED DESCRIPTION

[0041] In order to verify the efficacy of the Shenqi Tongmai preparation of the present invention, the inventor conducted a pharmacodynamic test study. It should be noted that the drugs selected for the pharmacodynamic test study of the present invention are drugs obtained by the representative formula of the present invention and its preparation method. The tests and results involved in the drugs obtained by other formulas and preparation methods included in the present invention are limited to the length of the article and are not listed here one by one.

[0042] Example 1: Preparation of Shenqi Tongmai Tablets

[0043] Formula:

[0044]

[0045] The preparation method is as follows:

[0046] A) Take earthworm and scorpion, and crush them into particles smaller than 100 μm;

[0047] B) Raw Astragalus, Atractylodes macrocephala, Cyathula officinalis, Coix seeds, Angelica sinensis, and Codonopsis pilosula were extracted once with 65% ethanol for 1.5 hours, filtered, and the filtrate was used to recover ethanol and concentrated under reduced pressure to an extract with a relative density of 1.05; the filtered residue was then extracted once with 2 times water, filtered, and the water extract was used for standby use;

[0048] C) extracting volatile oil from honeysuckle, salvia miltiorrhiza, liquorice and millettia spatholobi with water, boiling the remaining liquid and residues with 1 / 2 amount of earthworm and scorpion in water for 1 time, boiling for 1.5 hours each time, adding ethanol to the filtrate and the water extract in step B to make the alcohol content reach 40%, standing at room temperature for 24 hours, recovering ethanol from the filtrate, concentrating to an extract with a relative density of 1.15, and setting aside;

[0049] D) mixing the remaining 1 / 2 amount of earthworms and scorpions with the alcohol extract of step B and the water extract of step C, and preparing the mixture into tablets directly or by adding pharmaceutically acceptable excipients through conventional procedures.

[0050] Example 2: Preparation of Shenqi Tongmai Tablets

[0051] Formula:

[0052]

[0053] The preparation method is as follows:

[0054] A) Take earthworm and scorpion, and crush them into particles smaller than 100 μm;

[0055] B) Raw Astragalus, Atractylodes macrocephala, Cyathula officinalis, Coix seeds, Angelica sinensis, and Codonopsis pilosula were extracted twice with 85% ethanol, each time for 3.5 hours, filtered, the filtrate was used to recover ethanol, and the extract was concentrated under reduced pressure to a relative density of 1.10; the filtered residue was then extracted twice with 5 times water, filtered, and the water extract was used for later use;

[0056] C) extracting volatile oil from honeysuckle, salvia miltiorrhiza, liquorice and millettia spatholobi with water, boiling 1 / 2 amount of earthworm and scorpion in the remaining liquid and dregs with water for 2 times, boiling for 3.5 hours each time, adding ethanol to the filtrate and the water extract in step B to make the alcohol content reach 50%, standing at room temperature for 48 hours, recovering ethanol from the filtrate, concentrating to an extract with a relative density of 1.20, and setting aside;

[0057] D) mixing the remaining 1 / 2 amount of earthworms and scorpions with the alcohol extract of step B and the water extract of step C, and preparing the mixture into tablets directly or by adding pharmaceutically acceptable excipients through conventional procedures.

[0058] Example 3: Preparation of Shenqi Tongmai Tablets

[0059]

[0060] The preparation method is as follows:

[0061] A) Take earthworm and scorpion, and crush them into particles smaller than 100 μm;

[0062] B) Raw Astragalus, Atractylodes macrocephala, Cyathula officinalis, Coix seeds, Angelica sinensis, and Codonopsis pilosula were extracted twice with 75% ethanol, each time for 2.5 hours, filtered, the filtrate was used to recover ethanol, and concentrated under reduced pressure to an extract with a relative density of 1.08; the filtered residue was then extracted twice with 3 times water, filtered, and the water extract was used for later use;

[0063] C) extracting volatile oil from honeysuckle, salvia miltiorrhiza, liquorice and millettia spatholobi with water, boiling 1 / 2 amount of earthworm and scorpion in the remaining liquid and dregs with water for 2 times, boiling for 2.5 hours each time, adding ethanol to the filtrate and the water extract in step B to make the alcohol content reach 45%, standing at room temperature for 48 hours, recovering ethanol from the filtrate, concentrating to an extract with a relative density of 1.18, and setting aside;

[0064] D) mixing the remaining 1 / 2 amount of earthworms and scorpions with the alcohol extract of step B and the water extract of step C, and preparing the mixture into tablets directly or by adding pharmaceutically acceptable excipients through conventional procedures.

[0065] Example 4: Preparation of Shenqi Tongmai Tablets

[0066]

[0067] The preparation method is the same as Example 3.

[0068] Example 5: Preparation of Shenqi Tongmai Pills

[0069]

[0070] The preparation method is the same as that in Example 3, and excipients are added according to conventional processes to prepare pills.

[0071] Example 6: Preparation of Shenqitongmai External Ointment

[0072]

[0073] The preparation method is the same as that of Example 3, and auxiliary materials are added according to conventional processes to prepare the product for external use.

[0074] Comparative Example 1: Preparation of Shenqi Tongmai Tablets

[0075]

[0076] The preparation method is the same as Example 3.

[0077] Comparative Example 2: Preparation of Shenqi Tongmai Tablets

[0078]

[0079] The preparation method is the same as Example 3.

[0080] Comparative Example 3: Preparation of Shenqi Tongmai Tablets

[0081]

[0082] The preparation method is the same as Example 3.

[0083] Comparative Example 4: Preparation of Shenqi Tongmai Tablets

[0084]

[0085] The preparation method is the same as Example 3.

[0086] Comparative Example 5: Preparation of Shenqi Tongmai Tablets

[0087]

[0088] The preparation method is the same as Example 3.

[0089] Comparative Example 6: Preparation of Shenqi Tongmai External Cream

[0090]

[0091] The preparation method is as follows:

[0092] A) Take earthworm and scorpion, and crush them into particles smaller than 100 μm;

[0093] B) Raw Astragalus, Atractylodes macrocephala, Cyathula officinalis, Coix seeds, Angelica sinensis and Codonopsis pilosula were extracted twice with 75% ethanol, each time for 2.5 hours, filtered, ethanol was recovered from the filtrate, and concentrated under reduced pressure to an extract with a relative density of 1.08; set aside;

[0094] C) decocting honeysuckle, salvia miltiorrhiza, liquorice and millettia spatholobi with water for 2 times, each time for 2.5 hours, adding ethanol to the filtrate and the water extract in step B to make the alcohol content reach 45%, standing at room temperature for 48 hours, recovering ethanol from the filtrate, concentrating to an extract with a relative density of 1.18, and setting aside;

[0095] D) mixing the earthworm, scorpion, the alcohol extract of step B and the extract of step C, and preparing an external cream directly through conventional procedures or by adding a pharmaceutically acceptable excipient.

[0096] Comparative Example 7: Preparation of Shenqi Tongmai External Cream

[0097]

[0098] The preparation method is as follows:

[0099] A) Pheretima, scorpion, raw astragalus, white atractylodes, cyathula, coix seed, angelica, and codonopsis were extracted twice with 75% ethanol, each time for 2.5 hours, filtered, the filtrate was used to recover ethanol, and the extract was concentrated under reduced pressure to a relative density of 1.08; the filtered residue was then extracted twice with 3 times water, filtered, and the water extract was used for standby use;

[0100] B) extracting volatile oil from honeysuckle, salvia miltiorrhiza, liquorice and millettia spatholobi with water, decocting the remaining medicinal solution and medicinal residues with water for 2 times, decocting for 2.5 hours each time, adding ethanol to the filtrate and the aqueous extract in step B to make the alcohol content reach 45%, standing at room temperature for 48 hours, recovering ethanol from the filtrate, concentrating to an extract with a relative density of 1.18, and setting aside;

[0101] C) mixing the water extract with the alcohol extract of step B and the water extract of step C, and preparing an external cream directly or by adding a pharmaceutically acceptable excipient through conventional procedures.

[0102] 1. Pharmacological Experiments - Experiments on the Wound Surface of Mice Using the Preparation of the Invention and the Results

[0103] 1. Materials

[0104] 1.1 Animals:

[0105] Several SPF Kunming mice, 18-22 g, experimental animal license number: SYXK(Lu)20180004, were adaptively raised for 7 days before the experiment.

[0106] 1.2 Test substances

[0107] 1.2.1 Drugs

[0108] The cream was prepared according to the prescription and preparation method of Example 6 and Comparative Examples 6-7 of the present invention.

[0109] MEBO group (National Medicine Standard No. Z20000004).

[0110] 1.2.2 Usage

[0111] The ointment of the present invention is for external use. Apply to the wound surface or the surface to be healed (the thickness of the foam is 1-2 cm, and the thickness of the cream is thinner than 1 mm), and replace the new medicine every 2 to 4 hours. Before changing the medicine, wipe off the foam or liquid remaining on the wound surface. Expose the wound surface and then continue to apply the medicine.

[0112] Moist burn ointment group: For external use. Apply this product to burn, scald, scald and other wounds (thickness less than 1mm), and change the new medicine every 2 to 4 hours. Before changing the dressing, wipe off the medicine and liquefied substances remaining on the wound. Expose the wound surface and then continue to apply the medicine.

[0113] 2. Modeling, grouping and medication

[0114] 2.1 Modeling method

[0115] On the first day, the mouse back was depilated, with a depilation area of ​​about 3cm×4cm. The mice were observed for 24 hours to ensure that there were no abnormalities such as redness, swelling, inflammation, and damage at the depilation site. On the second day, at room temperature, the mice were anesthetized intraperitoneally, and placed in a prone position with their limbs stretched and fixed on the operating table to make the back skin flat. The hair on the back of the mice was pushed away, with a range of about 6cm, which was larger than the expected burn area. A 20g weight was placed in a container and heated to boiling for 10 minutes. After the mouse experimental area skin was disinfected with 75% ethanol, the heated weight was quickly placed on the mouse skin with a slight pressure to form a shallow second-degree burn model with a burn area of ​​about 2cm×2cm. The local skin after the burn was porcelain white, the epidermis was wrinkled, and local skin ulceration was common. The normal area and the burn site were clearly distinguishable, and subcutaneous tissue edema was obvious within 4 hours of the burn. One mouse was randomly selected to take its wound tissue for pathological histological examination. Microscopically, it was found that the epidermis in the burned area of ​​the mouse disappeared, and there was cell infiltration in the entire epidermis and the superficial dermis, which was consistent with superficial second-degree burns.

[0116] 2.2 Grouping and Dosing

[0117] Fifty mice with successful modeling were randomly divided into a model group, a MEBO group, an Example 6 group, and a Comparative Example 6-7 group, 10 mice each.

[0118] In each experimental group: the moist burn ointment group, the Example 6 group, and the comparative examples 6-7, the corresponding drugs were applied to the application area according to the above-mentioned use method, and the model group was applied with 0.9% saline. The drugs were administered continuously for 14 days.

[0119] 3. Observation indicators

[0120] 3.1 Lesion healing rate The local lesion area was measured by the tracing and weighing method on the 7th, 10th and 14th days after treatment. The lesion range was first marked with a transparent film, and then the transparent film corresponding to the size of the lesion area was cut and weighed, and the mass was used instead of the area.

[0121]

[0122] 3.2 Statistical methods

[0123] SPSS 17.0 software was used for data processing. The data were expressed as mean ± standard deviation. The data of each group were tested for normal distribution and homogeneity of variance, which met the requirements of Pearson's test, and one-way analysis of variance was used for comparison between groups. P < 0.05 was considered statistically significant.

[0124] 4. Results and Conclusions

[0125] On the 7th, 10th and 14th days after treatment, the skin lesions of Example 6 of the present invention, the comparative examples 5-6 groups and the moist burn ointment group were repaired to varying degrees. The longer the time, the better the healing. Among them, the healing effect of Example 6 of the present invention group was the best.

[0126] Compared with the model group, the healing rates of mice in each drug-treated group on the 7th, 10th and 14th days of treatment were higher, and the differences were statistically significant (P < 0.01), further indicating that the modeling of the present invention was successful; compared with the moist burn ointment group, the skin lesion healing rate of mice in the 6th group of the present invention was higher, and the difference was statistically significant (P < 0.05).

[0127] Table 1 Comparison of skin lesion healing rates in each group of mice ( n=10)

[0128] Group Treatment Day 7 Treatment day 10 Treatment day 14 Model Group 32.34±3.11 47.47±5.83 58.27±7.11 MEBO <![CDATA[42.43±5.82 & ]]> <![CDATA[57.74±6.14 & ]]> <![CDATA[74.92±8.19 & ]]> Example 6 <![CDATA[44.56±6.11 & ]]> <![CDATA[63.09±6.72 &# ]]> <![CDATA[84.33±7.75 &# ]]> Comparative Example 6 39.67±3.40 52.36±7.89 64.77±8.28 Comparative Example 7 38.33±5.71 53.46±5.11 65.98±6.42

[0129] Note: Compared with the model group, * P<0.05, & P < 0.01;

[0130] Compared with moist burn ointment, # P<0.05.

[0131] 2. Antibacterial test of the pharmaceutical composition of the present invention

[0132] 2.1 Materials and methods

[0133] 1) Bacterial species and culture medium: Sabouraud medium, Staphylococcus aureus, Candida albicans, Escherichia coli and Pseudomonas aeruginosa

[0134] Bacteria (the above strains were purchased through commercial channels).

[0135] 2) Drug: a drug composition prepared according to the raw materials and methods of Examples 3 to 4 and Comparative Examples 4 to 5 of the present invention (raw drug before adding excipients).

[0136] 3) Method: Prepare Sabouraud culture medium and put it into a conical flask, sterilize it with wet heat. Take 1 ml of each of Examples 3-4 of the present invention, Comparative Examples 4-5 and sterilized physiological saline 1:20 dilution by aseptic method and inject them into sterilized plates, 10 plates for each dosage group (plate diameter is 10 cm), and then pour into Sabouraud culture and inoculate the above four bacteria (bacterial mass with a diameter of about 0.5 mm). Incubate at 37°C for 6 days. Observe the growth state of the colony and whether there is growth of foreign bacteria, measure the colony diameter (mm) and compare it with the blank group.

[0137] Table 2 Antibacterial effect of each embodiment group ( n=10)

[0138]

[0139] Note: Compared with Comparative Example 41, * P<0.05, & P < 0.01;

[0140] Compared with Comparative Example 44, # P<0.05, @ P < 0.01;

[0141] As can be seen from Table 2 above, Examples 3-4 of the present invention have obvious antibacterial effects, and have obvious inhibitory or killing effects on pathogenic bacteria such as Staphylococcus aureus, Candida albicans, Enterobacter and Pseudomonas aeruginosa.

[0142] 2. Pharmacodynamics Experiment:

[0143] 1. Clinical research: This study selected 120 patients with diabetic ulcers of qi deficiency and blood stasis type who met the criteria from the outpatient or inpatient surgery department of Jinan Hospital of Traditional Chinese Medicine from July 2021 to July 2023, and randomly divided them into 5 groups. There was no significant difference in gender, age and wound area among the groups (P>0.05).

[0144] 2. Grouping and Dosing

[0145] Divided into experimental group A group, experimental group B group, experimental group C group, experimental group D group, positive control group, 24 examples in each group, take the tablets of embodiment 1 and comparative example 1-3 respectively, 6 tablets per day in each group, twice fasting warm in the morning and evening. 1 week is a course of treatment, 4 consecutive courses of treatment, positive control group (national medicine standard Z32020535 Tongsai Mai tablets, taken according to the instructions), according to the patient's blood sugar level and the actual situation of the patient, select appropriate hypoglycemic drugs, oral hypoglycemic drugs or subcutaneous injection of insulin treatment, so that fasting blood sugar is maintained below 8.0mmol / L. Each group is treated with local normal saline as needed.

[0146] Diagnostic criteria

[0147] 3.1 According to the diagnostic criteria of the Chinese Association of Traditional Chinese Medicine's "Guidelines for the Diagnosis and Treatment of Diabetic Foot in China" (2020 edition):

[0148] (1) Patients with type 2 diabetes who have limb vascular and / or neuropathy and / or infection;

[0149] (2) Patients with type 2 diabetes have clinical manifestations and signs of wet gangrene or dry gangrene in their limbs and meet the criteria for grade 1 to 5 gangrene;

[0150] (3) ABI index <0.9 at rest;

[0151] (4) Ultrasound color Doppler examination shows limb vascular stenosis and reduced limb blood supply leading to ischemia or gangrene;

[0152] (5) Patients with vascular stenosis or obstruction confirmed by angiography, CTA, or MRA, and clinical manifestations;

[0153] (6) Electrophysiological examination showed slowed peripheral nerve conduction velocity or abnormal changes in electromyography and somatosensory evoked potentials;

[0154] (7) X-ray examination shows osteoporosis, decalcification, bone destruction, osteomyelitis or joint lesions, hand and foot deformities, and Charcot joint changes;

[0155] The diagnosis can be confirmed if both (1) and (2) above are met, plus any one of (3)-(7).

[0156] 3.2 TCM diagnostic criteria

[0157] Refer to the diagnostic criteria for gangrene in the "Standards for Diagnosis and Efficacy of TCM Diseases and Syndromes" issued by the State Administration of Traditional Chinese Medicine in 2017:

[0158] (1) It often occurs on one or both sides of the lower limbs. The patient has a history of diabetes for many years, cold weather, trauma, long-term heavy smoking, etc. At first, the toes and fingers are cold and painful, and the lower limbs are sore, numb, and swollen. The pain worsens when walking for a long time and is relieved when resting. It is intermittent claudication and the Fuyang pulse is weakened. Subsequently, the pain becomes persistent, the skin of the extremities becomes cold, the skin becomes dark red or bluish purple, the skin becomes dry, the hair falls off, the toenails become deformed and thickened, the muscles atrophy, and the Fuyang pulse disappears. Then necrosis occurs, the pain is severe, the patient cannot sleep all night, and sits with his knees hugged. When the ulcer is infected, wet necrosis occurs, the extremities become red, swollen, hot and painful, and the whole body has a fever;

[0159] (2) Blood stasis type: The affected toe prefers warmth to cold, is numb, sore, swollen and painful, and is in severe pain when exposed to cold. The toe has difficulty walking, feels heavy and weak, and the pain worsens with more walking. The skin color is dark red or cyanotic. If the toe ulcers, the pain continues to worsen, and the toe cannot fall asleep all night. The tongue is dark red or has ecchymosis, the tongue coating is white, the pulse is stringy or astringent, and the Fuyang pulse disappears.

[0160] 3.3 Classification Standards

[0161] The most classic diabetic foot grading system, Wagner grading, was used.

[0162] Level 0: There are risk factors for foot ulcers, but no ulcers currently

[0163] Grade 1: Superficial foot ulcers with no signs of infection and prominent neuropathic ulcers

[0164] Grade 2: Deeper ulcers, often with soft tissue infection, without osteomyelitis or deep abscesses

[0165] Grade 3 deep ulcer with abscess or osteomyelitis

[0166] Grade 4 localized gangrene (toes, heels, or forefoot), characterized by ischemic gangrene, often with neuropathy

[0167] Grade 5 gangrene of the entire foot

[0168] 3.4 Inclusion criteria

[0169] (1) Meet the diagnostic criteria for type 2 diabetes; (2) Meet the Western medical diagnostic criteria for diabetic foot ulcers;

[0170] (3) Meet the diagnostic criteria of blood stasis in traditional Chinese medicine; (4) Wagner classification is 1-3;

[0171] (5) ankle-brachial index (ABI) ≥ 0.4; (6) age 18-80 years, regardless of gender;

[0172] 3.5 Exclusion criteria

[0173] (1) Severe limb ischemia requiring emergency vascular bypass or interventional treatment;

[0174] (2) Those with local necrosis or severe infection of the limbs who need amputation;

[0175] (3) ankle-brachial index (ABI) < 0.4;

[0176] (4) Cancerous, tuberculous and other specific ulcers;

[0177] (5) Patients with severe illness and unstable vital signs;

[0178] (6) Severe malnutrition;

[0179] (7) Patients with comorbid mental illness, alcoholism or drug abuse;

[0180] (8) Those with contraindications to ozone therapy such as hyperthyroidism, hypotension, hypocalcemia, hypoglycemia, ozone allergy, citrus allergy (for those using sodium citrate), coagulation disorder (such as hemophilia), and favism;

[0181] (9) Patients who are also participating in other trials;

[0182] 3.6 Rejection and shedding criteria

[0183] (1) Mistakes in inclusion, not meeting the inclusion criteria;

[0184] (2) those with poor compliance and unable to follow the prescribed treatment plan;

[0185] (3) Data are missing and effectiveness and safety cannot be judged;

[0186] (4) Severe adverse reactions to the drug occur and the trial must be terminated;

[0187] (5) The condition worsens or death occurs, leading to the termination of the study;

[0188] 4. Clinical observation indicators

[0189] The wound area was observed at 0, 1, 2, 3 and 4 weeks of treatment; serum (TNF-a, IL-6, high-sensitivity C-reactive protein (hs-CRP)) levels were detected at 4 weeks of treatment.

[0190] 5. Efficacy evaluation criteria

[0191] Referring to the clinical research guidelines formulated in the "Guidelines for Clinical Research of New Chinese Medicines (2022)", combined with the clinical characteristics of diabetic foot ulcers, the clinical efficacy evaluation criteria are formulated as follows:

[0192] Cured: The wound is basically healed, clinical symptoms and signs basically disappear, and the wound syndrome score index is ≥90%.

[0193] Significantly effective: The wound area (depth) is significantly reduced, the local pain of the wound disappears or is significantly alleviated, the secretion is significantly reduced, the granulation tissue is fresh, and there is no redness or swelling around it. 60%≤Wound symptom score index<90%.

[0194] Effective: The wound area (depth) is reduced, the local pain of the wound is relieved, the secretion is reduced, the granulation tissue is fresh, and the surrounding redness and swelling are reduced. 30%≤wound symptom score index<60%.

[0195] Ineffective: The wound area (depth) did not shrink or increased, the local pain of the wound did not decrease or worsened, the secretion did not decrease, and the wound condition did not improve. The wound symptom score index <30%.

[0196] Results Analysis

[0197] Figure 1 The figures show the wound healing results of Example 1 and Comparative Examples 1-3. It can be seen that Example 1 has the best therapeutic effect, and the number of individuals who were cured and showed significant effects is much greater than that of the other examples.

[0198] Figure 2 The figure shows the changes in wound area of ​​Example 1 and Comparative Examples 1-3. It can be seen that there were no significant changes between the groups in weeks 0-2. The treatment effects began to show significant stratification after 3 weeks of treatment. The treatment effect of Example 1 was significantly better than that of Comparative Examples 1-3.

[0199] Figure 3 The figure shows the comparison of wound surface area of ​​Example 1 and Comparative Examples 1-3 after 3 weeks of treatment. It can be seen that there are significant differences between the example and the other comparative examples.

[0200] Figure 4 The figure shows the comparison of wound surface area of ​​Example 1 and Comparative Examples 1-3 after 4 weeks of treatment. It can be seen that there is a significant difference between the example and the other comparative examples, and the difference in treatment effect is significantly widened.

[0201] Figure 5 Shown are the changes in TNF-a in Example 1 and Comparative Examples 1-3.

[0202] Figure 6 Shown are the changes in IL-6 in Example 1 and Comparative Examples 1-3.

[0203] Figure 7 It shows the changes of high-sensitivity C-reactive protein in Example 1 and Comparative Examples 1-3.

Claims

1. Use of a Shenqi Tongmai preparation in the preparation of a drug for preventing and treating diabetic neuropathy, wherein the Shenqi Tongmai preparation is prepared from the following Chinese medicines in parts by weight: 10-70 weight parts of raw astragalus 5-30 weight parts of white atractylodes 8-40 weight parts of Sichuan cyathula Coix seeds 10-60 weight parts Angelica sinensis 10-50 weight parts Salvia miltiorrhiza 5-30 weight parts Millettia reticulata 5-30 parts by weight Honeysuckle 5-30 parts by weight Codonopsis pilosula 10-50 parts by weight 5-30 parts by weight of earth dragon; 1-6 parts by weight of whole scorpion; 5-20 parts by weight of raw licorice.

2. The use according to claim 1, characterized in that The components of the Shenqi Tongmai preparation are: 30 weight parts of raw astragalus, 15 weight parts of atractylodes macrocephala, 18 weight parts of Cyathula officinalis 30 parts by weight of coix seed 20 parts by weight of angelica root 15 parts by weight of salvia miltiorrhiza Millettia reticulata 15 weight parts Honeysuckle 15 weight parts Codonopsis pilosula 30 weight parts 15 parts by weight of earthworms, 3 parts by weight of whole scorpions, 10 parts by weight of raw licorice.

3. The use according to any one of claims 1 to 2, characterized in that: The diabetes is type 1 diabetes or type 2 diabetes.

4. The use according to claim 3, characterized in that The diabetic neuropathy is painful diabetic peripheral neuropathy and diabetic lower limb vascular neuropathy.

5. The use according to claim 4, characterized in that The diabetic lower limb vascular neuropathy is diabetic foot.

6. The use according to claim 1, characterized in that The Shenqi Tongmai preparation is an oral preparation.

7. The use according to claim 6, characterized in that The oral preparation is one of decoction, pill, tablet, mixture, capsule, granule, powder or paste.

8. The use according to claim 1, characterized in that The preparation method of the Shenqi Tongmai preparation is: A. Take earthworms and scorpions, and crush them into particles less than 100μm; B. Add 65-85% ethanol to extract raw Astragalus, Atractylodes macrocephala, Cyathula officinalis, Coix seeds, Angelica sinensis, and Codonopsis pilosula for 1-2 times, each time for 1.5-3.5 hours, filter, recover ethanol from the filtrate, and concentrate under reduced pressure to an extract with a relative density of 1.05-1.10; then take the filtered residue and add 2-5 times water to extract 1-2 times, filter, and set aside the water extract; C. Extract volatile oil from honeysuckle, salvia miltiorrhiza, liquorice and millettia reticulata with water, add 1 / 2 amount of earthworm and scorpion to the remaining liquid and residue, and boil for 1-2 times, each time for 1.5-3.5 hours, add ethanol to the filtrate and the water extract in step B to make the alcohol content reach 40-50%, stand at room temperature for 24-48 hours, recover ethanol from the filtrate, concentrate to an extract with a relative density of 1.15-1.20, and set aside; D. Evenly mix the remaining 1 / 2 amount of earthworms and scorpions with the alcohol extract of step B and the water extract of step C, and prepare a clinically acceptable dosage form directly or by adding pharmaceutically acceptable excipients through conventional procedures.

9. The use according to claim 8, characterized in that The preparation method of the Shenqi Tongmai preparation is: A. Take earthworms and scorpions, and crush them into particles less than 100μm; B. Extract raw Astragalus, Atractylodes macrocephala, Cyathula officinalis, Coix seeds, Angelica sinensis and Codonopsis pilosula with 75% ethanol for 2 times, each time for 2.5 hours, filter, recover ethanol from the filtrate, and concentrate under reduced pressure to an extract with a relative density of 1.08; then extract the residue after filtration with 3 times water for 2 times, filter, and keep the water extract for later use; C. Extract volatile oil from honeysuckle, salvia miltiorrhiza, liquorice and millettia reticulata with water, add 1 / 2 of the prescribed amount of earthworm and scorpion to the remaining liquid and residue, and boil twice, each time for 2.5 hours, add ethanol to the filtrate and the water extract in step B to make the alcohol content reach 45%, stand at room temperature for 48 hours, recover ethanol from the filtrate, concentrate to an extract with a relative density of 1.18, and set aside; D. Evenly mix the remaining 1 / 2 amount of earthworms and scorpions with the alcohol extract of step B and the water extract of step C, and prepare a clinically acceptable dosage form directly or by adding pharmaceutically acceptable excipients through conventional procedures.

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