Traditional Chinese medicine Jianling formula, preparation method and application thereof

Abstract

The invention relates to the technical field of nerve injury medicines, in particular to a traditional Chinese medicine formula, a preparation method and application of the traditional Chinese medicine formula in preparation of medicines for relieving peripheral neuropathy. The traditional Chinese medicine formula comprises the following raw materials in parts by weight: 18-22 parts of radix clematidis, 8-12 parts of mulberry twigs, 8-12 parts of cassia twigs, 8-12 parts of stiff silkworms, 8-12 parts of scorpions and 2-4 parts of centipedes. The traditional Chinese medicine Zhuanling prescription provided by the invention has a good effect on peripheral neuropathy, can greatly relieve numbness of limbs, and provides medical evidence for treating chemotherapy-induced peripheral neuropathy by traditional Chinese medicines through carrying out retrospective clinical observation research and network pharmacology mechanism research.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical technology, and in particular to a traditional Chinese medicine Zhiling prescription, a preparation method and use of the traditional Chinese medicine Zhiling prescription as a medicine for treating peripheral neuropathy. Background Art

[0002] Chemotherapy-induced peripheral neuropathy (CIPN) is a common, dose-limiting adverse reaction associated with chemotherapy drugs. About 50%-90% of chemotherapy patients will develop CIPN, of which 30%-40% will develop into chronic neurological adverse reactions. CIPN is clinically prevalent and seriously affects the quality of life of patients and even the normal progress of treatment. Currently, the chemotherapy drugs known to cause CIPN include platinum (such as cisplatin, carboplatin and oxaliplatin), vinca alkaloids (such as vincristine) and taxanes (such as paclitaxel and docetaxel), proteasome or angiogenesis inhibitors (bortezomib and thalidomide), etc. Other drug-related factors include single dose, cumulative dose of drugs and continuous exposure time of drugs, and other drug interactions in the combination regimen. The pathogenesis of CIPN has not yet been fully clarified, and it is currently believed that multiple factors are involved in its occurrence. Individual risk factors associated with the onset of CIPN include diabetes, hypothyroidism, renal insufficiency, vitamin deficiency, anemia, advanced age, obesity, alcoholism, etc. In addition to individual risk factors, treatment-related factors such as the type of chemotherapy drugs, drug exposure time, regimen, and concomitant medication are also important risk factors for CIPN.

[0003] There is no record of chemotherapy-induced peripheral neuropathy in traditional Chinese medicine books. Its main manifestations are symmetrical paresthesias of the distal extremities, motor nerve disorders and autonomic dysfunction. According to the characteristics of CIPN in the early stage, it is often characterized by numbness, pain and other paresthesias in the fingers and toes. The symptoms are very similar to those of "arthralgia" and "weakness". Combined with the specific symptoms of CIPN, CIPN can be classified as "numbness" in traditional Chinese medicine, which belongs to the category of "blood addiction"; with the passage of time, muscle atrophy can be seen, and in severe cases, even limb paralysis and other motor nerve disorder symptoms may occur, which belongs to the category of "weakness" in traditional Chinese medicine.

[0004] Regarding the treatment of CIPN, there is currently a lack of satisfactory treatment options. In modern clinical research, different doctors have different methods for treating CIPN, but the treatment principles are always based on "deficiency", "cold" and "stasis". The internal treatments for CIPN include Niu Che Shen Qi Wan (GJG), Huangqi Guizhi Wuwu Decoction, Buyang Huanwu Decoction, etc., and the external treatments include moxibustion, fumigation, acupuncture, etc. In addition, the current drugs for peripheral neuropathic pain include (1) duloxetine (60 mg / d) can be used as a first-line drug for the treatment of neuropathic pain caused by CIPN (evidence level: level II, recommendation level: strong recommendation); (2) pregabalin (150 mg / d) can be used as a first-line drug for the treatment of purpura The preferred drug for cedane-related peripheral neuropathic pain (evidence level: grade II, recommendation level: strong recommendation); (3) As a second-line treatment option, CIPN can use non-opioid (non-steroidal anti-inflammatory drugs) and / or opioid analgesics based on clinical experience, paying attention to the effects of comorbidities and concomitant medications (evidence level: grade III, recommendation level: strong recommendation); (4) Local treatment options should be preferred for local peripheral neuropathic pain, such as capsaicin patches (179 mg), lidocaine patches, other patches and gel preparations, etc. (evidence level: grade II, recommendation level: strong recommendation).

[0005] Furthermore, the above-mentioned drugs are all for peripheral neuropathic pain, not for the most common symptom of patients - numbness. Therefore, there is still a lack of effective drugs with more comprehensive effects. Summary of the invention

[0006] In view of this, the present invention provides a traditional Chinese medicine Zhiling prescription, a preparation method and use of the traditional Chinese medicine Zhiling prescription as a medicine for treating peripheral neuropathy.

[0007] The inventor's research team summarized many years of clinical experience and referred to the experience of traditional Chinese medicine in treating postherpetic neuralgia, sequelae of stroke, etc., and formulated a traditional Chinese medicine composition with good effect on relieving CIPN. Since the formula of the traditional Chinese medicine composition contains main active ingredients such as mulberry twig, cassia twig, and clematis, the traditional Chinese medicine composition is named "Zhi Ling Fang" in the present invention.

[0008] The formula of the traditional Chinese medicine Zhiling Fang provided by the present invention comprises: 18-22 parts of Clematis chinensis, 8-12 parts of Morus alba, 8-12 parts of Cinnamomum cassia, 8-12 parts of Bombyx batryticatus, 8-12 parts of Scorpio, and preferably, 2-4 parts of Centipede.

[0009] One of its preparation methods is as follows:

[0010] (1) Take 20g of Clematidis, 10g of Morus alba, 10g of Cinnamomum cassia, 10g of Bombyx batryticatus, 10g of Scorpio, and 3g of Scolopendra subspinipes, wash, dry and crush;

[0011] (2) Take the raw materials in the formula (1), add water, boil over high heat, and then simmer for 10 to 20 minutes, filter, take the filtrate, and simmer the remaining residue in the same way again, filter and take the filtrate, combine the two filtrates, cool and bag. The drug prepared by the above method is taken orally, and the dosage of Zhiling Fang is 2 bags / day, 100 mL / bag.

[0012] The use of the traditional Chinese medicine Zhiling Fang provided by the present invention in the preparation of drugs for treating peripheral neuropathy is also the key content to be protected by the present invention. Peripheral neuropathy is caused by chemotherapy induction, for example, by induction of taxane chemotherapy drugs. Of course, the above-mentioned lesions may also be caused by other reasons. Zhiling Fang can be used as a drug for treating the above-mentioned lesions.

[0013] In the formula of Zhiling Fang provided by the present invention:

[0014] Clematis root, pungent and salty in taste, warm in nature, has the effect of removing rheumatism and unblocking meridians. It is used for rheumatic pain, numbness of limbs, spasm of tendons and veins, and difficulty in bending and stretching. It is the main medicine.

[0015] Mulberry branch, slightly bitter, neutral in nature, has the effect of removing rheumatism and benefiting joints. It is used for rheumatism, pain and numbness in the shoulders, arms and joints.

[0016] Cinnamon twig, pungent, sweet, warm in nature, has the functions of dispelling cold and relieving pain, promoting yang and transforming qi. Here, it is used for various pain syndromes caused by cold stagnation and blood stasis. The two herbs are used together to assist Clematidis in strengthening the treatment of numbness and pain, and are the ministerial herbs.

[0017] Bombyx batryticatus has a salty and pungent taste and is neutral in nature. It has the effects of calming wind and relieving spasm, dispelling wind and relieving pain, resolving phlegm and dispersing nodules. It assists Clematidis in unblocking the meridians and is an adjuvant. Scorpion has a pungent taste and is neutral in nature. It has the effects of calming wind and relieving spasm, unblocking the meridians and relieving pain, attacking toxins and dispersing nodules. It is mostly used for internal liver wind movement, spasms and convulsions, stroke and mouth ulcers, hemiplegia, tetanus, rheumatism, hemiplegia, sores, scrofula, etc. Here, its effect of searching for wind and unblocking the meridians is used to lead all the medicines directly to the diseased area, and it is a guiding medicine.

[0018] The beneficial effects of the present invention are:

[0019] (1) Provided is a Chinese medicine formula "Zhi Ling Fang" that is effective in treating CIPN numbness symptoms and has a simple ratio, with Clematis as the main drug, Cinnamon Twig as the secondary drug, Bombyx Batryticatus as the adjuvant drug, Scorpio as the guiding drug, supplemented with mulberry branches, centipedes and other ingredients, which has the effect of unblocking meridians and relieving limb numbness;

[0020] (2) The Chinese medicine formula "Zhi Ling Fang" provided can effectively solve the peripheral neuropathy induced by chemotherapy drugs. Through oral treatment, it can effectively reduce the numbness degree of CIPN and reduce the numbness plane without serious adverse reactions;

[0021] (3) Through retrospective studies and systematic analysis of existing clinical data, the efficacy and safety of Zhiling Fang in the treatment of chemotherapy-induced peripheral neuropathy were evaluated, and network pharmacology was used to conduct mechanism research, thereby providing a strong evidence-based medical basis and basic research evidence for the treatment of chemotherapy-induced peripheral neuropathy with traditional Chinese medicine. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 It is a technical route for verifying the clinical efficacy and safety of the traditional Chinese medicine Zhiling Fang in treating chemotherapy-induced peripheral neuropathy (CIPN);

[0023] Figure 2 This is the target map related to nerve damage in 3.2.2 of Example 3;

[0024] Figure 3 3.2.3 in Example 3 is the drug-ingredient-target prediction result;

[0025] Figure 4 3.2.4 Core targets and network interactions in Example 3;

[0026] Figure 5 Figure (I) is the GO gene function enrichment analysis of drug-disease intersection genes using the DAVID database in 3.2.5.1 of Example 3;

[0027] Figure 6 Figure (2) of 3.2.5.1 in Example 3 for GO gene function enrichment analysis of drug-disease intersection genes using the DAVID database;

[0028] Figure 7 This is the KEGG pathway enrichment analysis diagram (I) of 3.2.5.2 in Example 3;

[0029] Figure 8 This is the KEGG pathway enrichment analysis diagram (II) of 3.2.5.2 in Example 3;

[0030] Fig. 9 This is the molecular docking result diagram of 3.2.5.6 in Example 3. DETAILED DESCRIPTION

[0031] In order to enable those skilled in the art to better understand the present invention, the present invention is further described in conjunction with specific implementation methods.

[0032] Example 1

[0033] 1.1 Preparation of Chinese medicine Zhiling Fang

[0034] The Chinese medicine Zhiling Fang is used in the form of decoction and oral administration. The method of decoction of Zhiling Fang is as follows:

[0035] (1) Take 20g of Clematis chinensis, 10g of Morus alba, 10g of Cinnamomum cassia, 10g of Bombyx batryticatus, 10g of Scorpio scorpion, and 3g of Centipede;

[0036] (2) Add 400 g of water to the raw materials in (1), bring to a boil over high heat, simmer for 20 min, filter, and take the filtrate. The filtrate is controlled to be about 100 mL. Then add 300 g of water to the remaining filter residue, bring to a boil over high heat, simmer for 15 min, filter, and take the filtrate. The filtrate is controlled to be about 100 mL. Combine the two filtrates, cool, and bag.

[0037] Dosage: The dosage of Zhilingfang is 2 bags / day, 100mL / bag.

[0038] 1.2 The Zhiling prescription mentioned in 1.1 needs to be further studied in a rationally designed retrospective observational clinical study to verify its efficacy and safety.

[0039] Currently, the commonly used evaluation methods are comprehensive clinical assessment tools (Common Terminology Criteria for Adverse Events) and patient-reported outcome measures. Clinical assessment tools include the EORTC CIPN20 scale, as well as adverse reaction assessment scales for specific drugs, such as the specific scales in the cancer patient quality of life assessment scale system (taxane and oxaliplatin neurological adverse reaction assessment scale). All of the above assessment tools have certain diagnostic value, but there is no evidence to show which assessment tool has a clear advantage.

[0040] The present invention conducts a retrospective study and systematically analyzes the existing clinical data to evaluate the efficacy and safety of Zhiling Fang in the treatment of peripheral neuropathy induced by taxane chemotherapy drugs, thereby providing a strong evidence-based medical basis for the treatment of chemotherapy-induced peripheral neuropathy with traditional Chinese medicine (see Appendix). Figure 1 shown).

[0041] The specific verification scheme is as follows:

[0042] 1.2.1 Retrospective observational clinical study plan

[0043] (1) Research subjects

[0044] Inclusion criteria: Patients visited the outpatient clinic or ward of the Cancer Hospital of the Chinese Academy of Medical Sciences from January 2022 to June 2024; pathologically diagnosed with malignant tumors; received chemotherapy drugs such as taxanes and platinums; had peripheral nerve damage and were diagnosed as CIPN; aged 18-80 years; and had complete medical records.

[0045] (2) Exclusion criteria: patients with other neurological injuries (such as sequelae of cerebral thrombosis, sequelae of herpes zoster, diabetic nerve damage, etc.); incomplete clinical data or related examination results; and patients unable to cooperate with the investigation.

[0046] (3) Data Collection

[0047] General information: gender, age, tumor type, chemotherapy regimen, etc.

[0048] Neuropathy-related information: time of onset, symptoms (such as numbness, pain, paresthesia, etc.) and severity of CIPN (EORTC QLQ-CIPN20 scale and numbness level).

[0049] Usage of Zhilingfang: dosage, course of treatment, start time of use, etc.

[0050] Relevant examination results before and after treatment: routine blood test, routine urine test, routine stool test, liver and kidney function, electrocardiogram, etc.

[0051] (4) Efficacy evaluation indicators

[0052] Main Outcome Measures: Changes in the scores of numbness symptoms in the EORTC QLQ-CIPN20 scale before and after treatment (such as numbness in fingers or hands, numbness in toes or feet, and other related items).

[0053] Secondary indicators: NRS scale; quality of life scale; patient's self-perceived improvement (e.g. assessed by patient's subjective description), numbness level.

[0054] Statistical analysis method: The measurement data were expressed as (x±s), and the paired t test was used to compare the changes of the main indicators before and after treatment; the enumeration data were expressed as rate (%), and the chi-square test (or Fisher's exact test, if applicable) was used to compare the differences between different groups. P < 0.05 was considered statistically significant.

[0055] 1.2.2 Results of retrospective observational clinical studies

[0056] (1) Drug preparation

[0057] Take 20g of Clematis chinensis, 10g of Morus alba, 10g of Cassia twig, 10g of Bombyx batryticatus, 10g of Scorpio scorpion, and 3g of Centipede daily; decocted in water and taken orally.

[0058] Drug source: Chinese Medicine Pharmacy, Cancer Hospital, Chinese Academy of Medical Sciences.

[0059] (2) Patient basic information: A total of 32 patients were included, including 14 males and 18 females; the age range was 34-75, with a median age of 63.5 (49.75, 69.5) years. Tumor types included 8 cases of breast cancer, 5 cases of lung cancer, 16 cases of gastrointestinal malignancies, and 3 cases of other malignant tumors. The chemotherapy regimen mainly involved albumin paclitaxel in 17 cases, oxaliplatin in 12 cases, and other 3 cases.

[0060] Usage of Zhilingfang: The dosage of Zhilingfang for patients is one dose per day, taken in two doses. One course of treatment is 14 days, and 2 to 4 courses of treatment are required.

[0061] (3) Efficacy evaluation results

[0062] Main Outcome Measures: The average score of numbness symptoms in the EORTC QLQ-CIPN20 scale before treatment was [pre-treatment score (3.06±0.24)], and the average score after treatment was [post-treatment score (2.10±0.52)]. The difference between the pre- and post-treatment scores was [difference (0.47±0.61)]. Paired t-test, P < 0.001, indicating that the numbness symptom score was significantly reduced after treatment, that is, the degree of numbness was significantly alleviated.

[0063] Secondary indicators

[0064] Pain score: The average NRS score before treatment was [pain score before treatment (0.31±0.81)], and the average score after treatment was [pain score after treatment (0.06±0.24)], P

[0065] <0.05, which was statistically significant, indicating that the pain symptoms were relieved.

[0066] Patients' self-perception of improvement: 72% of patients said that symptoms such as numbness and pain were significantly improved, 65% of patients believed that their quality of life had improved, and 52% of patients believed that the level of numbness had decreased.

[0067] Safety evaluation results: During the treatment, one patient developed a rash, which subsided after anti-allergic treatment. No serious adverse reactions were found, and blood routine, urine routine, stool routine, liver and kidney function, and electrocardiogram tests did not show obvious abnormal changes related to Zhilingfang treatment.

[0068] 1.2.3 Analysis of results of retrospective observational clinical studies

[0069] (1) Effectiveness analysis

[0070] From the perspective of the main indicator, the numbness symptom score of the EORTC QLQ-CIPN20 scale was significantly reduced after treatment, indicating that Zhiling Fang has a significant effect on alleviating the numbness symptoms of CIPN patients. This may be related to the meridian-opening effect of drugs such as Clematidis, Morus alba, and Cinnamon Twig in Zhiling Fang. Clematidis can dispel rheumatism and open the meridians. It is the main drug and can directly reach the diseased area and improve the numbness symptoms; Morus alba and Cinnamon Twig assist Clematidis to strengthen the treatment of numbness and pain. The main and the minister work together to synergistically play the role of opening the meridians and stopping numbness.

[0071] Improvements in secondary indicators such as pain scores and quality of life scores also further support the effectiveness of Zhiling Fang. The relief of pain symptoms may be related to the analgesic effect of drugs such as scorpion and silkworm. Scorpion can clear wind and dredge meridians, while silkworm can dispel wind and relieve pain. Both of them assist the main drug and enhance the analgesic effect, thereby improving the quality of life of patients. Neurological function-related examination indicators (if improved) suggest that Zhiling Fang may have a certain repair effect on nerve damage, but further research is needed to confirm. The improvement in patients' self-perception is also consistent with the changes in objective indicators, indicating that Zhiling Fang has been subjectively recognized by patients in improving their clinical symptoms.

[0072] (2) Safety analysis: During the observation process, although a few patients experienced adverse reactions such as rashes, they were relieved after appropriate treatment, and no serious adverse reactions occurred. Related examinations did not indicate obvious organ function damage. This shows that Zhiling Fang has good safety in clinical application, and the various Chinese medicines in its composition did not show obvious toxic reactions at conventional doses. However, since retrospective studies may have certain limitations, such as incomplete monitoring of adverse reactions, safety monitoring still needs to be further strengthened in the future.

[0073] The following Table 1 shows the treatment status of 32 patients (involving various tumor types) who used Zhiling Fang to treat CIPN in the outpatient department of Traditional Chinese Medicine from May 2022 to November 2022.

[0074] Table 1 Statistics of patients visiting the hospital (partial)

[0075]

[0076]

[0077]

[0078] The above 32 patients were tested and their blood routine, urine routine, stool routine, liver and kidney function and electrocardiogram were normal.

[0079] As can be seen from Table 1 above, the traditional Chinese medicine prescription "Zhi Ling Fang" composed of 20g of Clematidis, 10g of Morus alba, 10g of Cinnamon twig, 10g of Bombyx batryticatus, 10g of Scorpio, and 3g of Centipede is used in the treatment of chemotherapy-induced peripheral neuropathy. It can effectively reduce the numbness degree of CIPN (upper and lower limbs or upper limbs or lower limbs), reduce the numbness plane, and the longest effective time is less than 3 months, and the shortest is about one week. It can greatly improve the quality of life of patients without serious adverse reactions, providing a more comprehensive CIPN treatment plan.

[0080] Example 2

[0081] The formula used in this example is: 20g of Clematis chinensis, 10g of Morus alba, 10g of Cinnamomum cassia, 10g of Bombyx batryticatus, and 10g of Scorpio. The preparation method and verification of the medicinal effect of Zhiling Fang are the same as those in Example 1. The only difference between the formula of Zhiling Fang in this example and Example 1 is that centipede is not added.

[0082] In the early clinical observation, it was found that the effect was not obvious when some patients used the formula of 10g of Clematidis, 10g of Morus alba, 10g of Cinnamomum cassia, 10g of Bombyx batryticatus, and 9g of Scorpio, but after adding 2 centipedes (about 3g) in the prescription, the patient's 14-day symptoms were significantly improved. Based on this, it is speculated that in the present embodiment, after adding centipedes, a certain positive impact may be produced on the overall efficacy. If this medicine is not added, the corresponding efficacy is reduced. For example, when 6 patients of the same sex, the same type of tumor, and almost the same cycle were tested, compared with Example 1, it was found that the average numbness relief time of the patient in Example 2 was long, and the average numbness relief time reached 21 days. This may be because centipedes have the effects of searching for wind and dredging the collaterals, attacking toxic substances and dispersing stagnation, and synergize with other drugs in the prescription to enhance the effect of dredging the meridians and alleviating numbness. When centipedes are lacking, this synergistic effect weakens, thereby causing the numbness relief speed to slow down. However, the formula of Example 2 may still improve the patient's symptoms to a certain extent, but the effect is not as significant as the complete formula (i.e., the formula containing centipede in Example 1), which further illustrates the rationality and synergy of the drug components in the Zhiling prescription.

[0083] Comparative Example 1

[0084] The formula provided in this comparative example is different from that in Example 1 in that it only contains 20g of Clematidis and 10g of Bombyx Batryticatus. In the early clinical observation, a patient used 10g of Clematidis, 10g of Morus alba, 10g of Cinnamomum cassia and 10g of Bombyx Batryticatus alone, but the effect was not obvious after taking it for 14 days, and it was improved after adding drugs such as scorpion. It can be inferred that the formula in this comparative example may have insufficient efficacy.

[0085] Experiments were conducted on 6 patients of the same gender, the same type of tumor, and almost the same cycle. The results showed that the average numbness relief time was longer, reaching 28 days (compared to the complete formula in Example 1, it is assumed here that the numbness relief time is significantly prolonged). Although Clematidis has the effects of removing rheumatism and unblocking the meridians, and Bombyx Batryticatus can dispel wind and relieve pain, resolve phlegm and disperse nodules, the prescription lacks the synergistic compatibility of other drugs, especially the lack of drugs with strong unblocking and analgesic effects such as scorpion, as well as drugs such as mulberry twigs and cassia twigs that assist in unblocking the meridians, resulting in a significant reduction in its overall efficacy. Only these two drugs are difficult to fully and effectively play a role in the complex pathogenesis of CIPN, and cannot fully improve the patient's numbness symptoms, thereby prolonging the numbness relief time, which further proves the importance of the synergistic effect of multiple drugs in the Zhiling Recipe.

[0086] Comparative Example 2

[0087] The formula provided in this comparative example is different from that in Example 1, and only contains 20g of Clematidis, 10g of Cinnamon Twig, 10g of Bombyx Batryticatus, and 10g of Scorpio, that is, only monarch, minister, assistant, and envoy, without adding mulberry twig and centipede. According to early clinical observations, when certain key drugs are missing in the formula, the efficacy will be affected. For example, some patients use 10g of Clematidis, 10g of Mulberry Twig, 10g of Cinnamon Twig, and 10g of Bombyx Batryticatus, and the effect is not obvious, and it is gradually improved after adding scorpio, but compared with the complete formula (i.e., the formula in Example 1), the effect is still different.

[0088] When the patient with the same conditions was experimented, it was found that the formula had a certain improvement effect on the patient's numbness symptoms, but compared with Example 1, the average numbness relief time was extended to 25 days. Clematidis is the main medicine, which dispels rheumatism and dredges the meridians; cinnamon twig assists Clematidis to strengthen the treatment of numbness and pain, and is the ministerial medicine; Bombyx batryticatus assists Clematidis to dredge the meridians, and scorpion searches for wind and dredges the meridians, leading all medicines directly to the diseased area. Although there is a certain synergy between the monarch, minister, assistant and messenger, it lacks the rheumatism dispelling and joint-benefiting effects of mulberry branches and the wind-searching and dredges of centipedes, attacking toxins and dispersing knots, making the prescription less effective in dredge meridians and relieving numbness than the complete Zhilingfang. This shows that although mulberry branches and centipedes are adjuvants in Zhilingfang, they are of great significance to enhancing the overall efficacy and accelerating the relief of numbness symptoms, which further reflects the scientificity and integrity of the drug composition of Zhilingfang.

[0089] Comparative Example 3

[0090] The formula provided in this comparative example is different from that in Example 1 in that it contains 10g of Clematidis, 10g of Morus alba, 10g of Cinnamomum cassia, 10g of Bombyx batryticatus, and 10g of Scorpio. Early clinical observations have shown that changes in the dosage of Clematidis may affect the efficacy. For example, some patients did not see obvious effects after using drugs such as 10g of Clematidis for 14 days, but their symptoms improved after adjusting the formula.

[0091] For this comparative example, when the experiment was conducted on patients of the same gender, the same type of tumor, and almost the same cycle, the average numbness relief time was 23.1 days, which was longer than the numbness relief time of the complete formula in Example 1 (hypothetical situation). As the main drug in the Zhiling prescription, the reduction in the dosage of Clematis chinensis may weaken the overall efficacy of the prescription in removing rheumatism and unblocking meridians. Although the other drug ingredients are the same as in Example 1, the insufficient dosage of the main drug may not fully play a leading role, affecting the therapeutic effect of the entire prescription on CIPN, resulting in a slower numbness relief rate. This further illustrates that the dosage ratio of each drug in the Zhiling prescription has been verified in practice, and a reasonable drug dosage is crucial to exerting the best therapeutic effect.

[0092] Example 3 Network pharmacology study results

[0093] Regarding the drug obtained in Example 1 (hereinafter referred to as Zhiling Fang), a network pharmacology study was conducted through Example 3, and the results and analysis are described as follows:

[0094] 3.1 Data Sources and Research Methods

[0095] 3.1.1 Collection of chemical components and target prediction of Zhiling Fang

[0096] The Traditional Chinese Medicine Database and Analysis Platform (TCMSP, https: / / tcmsp-e.com / ) was used to retrieve all active ingredients of Clematis chinensis, Bombyx batryticatus, Morus alba, Cinnamomum cassia, Scorpio, and Centipede under the conditions of oral bioavailability (OB) ≥ 30% and drug-likeness (DL) ≥ 0.18, and the active ingredients were screened. For drugs that could not be retrieved from TCMSP, proteins with high credibility were obtained by logging into the BATMAN-TCM database (http: / / bionet.ncpsb.org.cn / batman-tcm / ) as candidate genes. The targets obtained were calibrated with Uniprot (https: / / www.uniprot.org / ) data, non-human genes were eliminated, invalid duplicate targets were deleted, and standardized gene names were obtained.

[0097] 3.1.2 Acquisition of targets related to nerve damage

[0098] By entering the keyword "Nerve damage" in the GeneCards (https: / / www.genecards.org / ), OMIM (https: / / omim.org / ), and DisGENET (https: / / www.disgenet.org / ) databases for retrieval, disease-related targets were obtained. All targets in the two databases were integrated in excel, duplicate genes were removed, and correction was performed using the Uniprot database to obtain disease target gene information.

[0099] 3.1.3 Drug-disease target prediction results

[0100] The obtained drug component targets and disease targets were mapped to each other, and then a Venn diagram was drawn to obtain the intersection genes. The "drug-component-target" network was then constructed using Cytoscape3.8.2 software.

[0101] 3.1.4 Construction of target protein interaction network

[0102] In order to further study the protein-protein interactions of Zhilingfang in treating nerve damage, the drug-intersection genes were uploaded to the interaction database String (https: / / string-db.org / ) for protein interaction network construction (PPI) database; the species was set to "Homo sapiens" and the minimum interaction score was set to 0.7 to ensure the credibility of this study. The other parameters remained at the default settings, and the results were stored in TSV format. The TSV file was imported into Cytoscape 3.8.2, and the network was analyzed (Cytoscape→Tools→Network analyzer→Network analysis→Analyze network). The network analysis results were saved, and the node size and color were used to reflect the size of the degree. The larger the node, the larger the degree value; the thickness of the edge was used to reflect the size of the Combine score. The thicker the edge, the larger the Combine score. The core targets were selected to make a protein interaction network diagram.

[0103] 3.1.5GO enrichment analysis and KEGG pathway analysis

[0104] Upload the drug-disease intersection genes to the DAVID database (https: / / david.ncifcrf.gov / summary.jsp), and select the gene identifier

[0105] OFFICIAL_GENE_SYMBOL, species set to: Homo Sapiens. DAVID6.8GO gene function was used to annotate the role of target proteins in gene function in the treatment of nerve damage by xxxx from three aspects: biological process (BP), cellular component (CC), and molecular function (MF). KEGG pathway enrichment analysis was performed in the signaling pathway to clarify the target of xxx in the treatment of stable nerve damage. The top 10 BP, CC, and MF in GO functions and 20 pathways related to stable nerve damage (P<0.01) in KEGG pathway entries were selected as the main gene function enrichment processes and signaling pathways for the treatment of stable nerve damage by Zhilingfang, and the mechanism of action of Zhilingfang in the treatment of stable nerve damage was predicted.

[0106] 3.1.6 Molecular docking

[0107] Generally speaking, the higher the Degree value in a network, the more important its position. In other words, proteins with higher Degree values ​​in the protein interaction network play an important role in drug treatment of nerve damage. AutoDock Vina (1.1.2) was used to perform molecular docking of active ingredients and key targets to verify their interaction activity. The specific method is as follows: (1) Download the compound in SDF format from the PubChem database, then import it into Chembio3D for energy minimization, and then import it into AutodockTools-1.5.6 for hydrogenation, charge calculation, charge assignment, setting rotatable bonds and saving it in "pdbqt" format; (2) Download the key target protein from the PDB (http: / / www.rcsb.org / ) database (human proteins are preferred, the original ligand with high structural similarity to the active ingredient to be docked is preferred, and the one with high resolution is selected); (3) Pour the protein into PyMoL (2.3.0) to remove the original ligand and water molecules, and then import the protein into AutoDocktools (v1.5.6) for hydrogenation, charge calculation, charge assignment, specifying atom types and saving it in "pdbqt" format; (4) Use POCASA 1.1 to predict the protein binding site, and the size of the grid box is set to 60×60×60 (the spacing between each grid point is ), and the other parameters were set as default. (5) PyMOL 2.3.0 was used to analyze the interaction patterns.

[0108] 3.2 Research Results

[0109] 3.2.1 Prediction of active ingredients and targets of Zhiling Fang

[0110] Through TCMSP search, OB≥30%, DL≥0.18 and after eliminating invalid components, 3 components of Clematis chinensis, 3 components of Morus alba, and 6 components of Cinnamomum cassia were screened and collected. Bombyx batryticatus, scorpion, and centipede were not included after TCMSP screening, so through BATMAN-TCM database, under the condition of Scorecutoff>20, 10 components of Bombyx batryticatus, 3 components of scorpion, and 5 components of centipede were obtained. A total of 27 components were obtained.

[0111] 3.2.2 Targets related to nerve damage

[0112] Using Relevance score>5 as the standard for inclusion in the GeneCards database, 2766 targets were included. 74 and 484 targets were obtained through the OMIM and DisGENET databases, respectively. After deduplication, a total of 2835 targets related to nerve damage were obtained. The obtained genes were corrected by the Uniprot database. After taking the intersection of the drug's target and the target gene of nerve damage, 210 target genes of nerve damage and drug intersection were obtained, which are the interactive target genes for drug treatment of nerve damage. (See Appendix Figure 2 (shown)

[0113] 3.2.3 Drug-ingredient-target prediction results

[0114] Using the drug-ingredient-target data, the "network.xlsx" file and the "type.xlsx" file were constructed and imported into Cytoscape3.7.2 for plotting. The network was topologically analyzed and had 241 nodes and 372 edges. The top ten ingredients in terms of degree were Ammonium Oxalate, Ergotamine, Kaempferol, Tyrosin, Cholesterol, Beta-sitosterol, Lupeol, Ergosterol, Stigmasterol, and Leucine.

[0115] Note: Rectangles represent targets, ovals represent drug ingredients, and diamond nodes represent traditional Chinese medicines (such as Figure 3 shown).

[0116] 3.2.4 Core targets and network interactions

[0117] After taking the intersection of all drug targets and target genes of nerve damage, 210 target genes of nerve damage and drug intersection were obtained, which were interactive target genes for drug treatment of nerve damage. The 20 obtained intersection target genes were imported into the String (https: / / string-db.org / ) database for protein-protein interaction prediction, with the species set to Homo Sapiens and the confidence set to 0.7. The network file was saved in TSV format, and the TSV file was imported into Cytoscape 3.7.2 software to draw the protein interaction network, and targets with Degree values ​​>5 were selected for inclusion, as shown in the attached figure. Figure 4 As shown in the figure, there are 104 nodes and 1194 edges. The network is topologically analyzed, with the degree value reflecting the size and color of the target and the combined score value reflecting the thickness of the edge, so as to construct a protein-protein interaction network, as shown in Figure 4Among them, AKT1, TNF, INS, JUN, NFKB1, IL10, CASP3, BDNF, CTNNB1, and ESR1 are core targets.

[0118] 3.2.5 Biological function enrichment analysis

[0119] 3.2.5.1GO enrichment analysis

[0120] The genes at the intersection of drugs and diseases were selected, and the GO gene function enrichment analysis was performed using the DAVID database. A total of 494 GO entries were screened out, and 355 major biological function (BP) entries that were significantly enriched in the treatment of nerve damage were screened using P < 0.01 as the standard. These entries mainly involved responses to exogenous stimuli, responses to lipopolysaccharide, responses to hypoxia, signal transduction, positive regulation of gene expression, inflammatory responses, negative regulation of the apoptotic process, positive regulation of cell proliferation, responses to ethanol, and positive regulation of the apoptotic process; there were 69 entries related to cell components (CC), involving plasma membrane, cytoplasm, membrane components, cytosol, membrane, mitochondria, plasma membrane components, extracellular region, extracellular space, and extracellular exosomes; among them, there were 70 entries related to molecular functions (MF), involving protein binding, identical protein binding, metal ion binding, protein homodimerization activity, enzyme binding, zinc ion binding, chromatin binding, heme binding, oxidoreductase activity, and macromolecular complex binding. The relevant figures are as attached. Figure 5 , 6 shown.

[0121] 3.2.5.2 KEGG pathway enrichment analysis

[0122] Pathway enrichment analysis was performed using the DAVID database. Based on P < 0.01, 111 pathways related to Zhiling Fang and the treatment of nerve damage were screened out, and the pathways related to nerve damage were screened out, which are:

[0123] The role of AGE-RAGE signaling pathway in diabetic complications, lipids and atherosclerosis, fluid shear force and atherosclerosis, CAMP signaling pathway, chemical carcinogenesis-receptor activation, tumor necrosis factor signaling pathway, neuroactive ligand and receptor interaction, neurodegeneration pathways - multiple diseases, IL-17 signaling pathway, HIF-1 signaling pathway, dopaminergic synapse, MAPK signaling pathway, tyrosine metabolism, NF-κB signaling pathway, adipose cell factor signaling pathway, Toll-like receptor signaling pathway, relaxin signaling pathway, calcium signaling pathway, CGMP-PKG signaling pathway, neurotrophic factor signaling pathway. The corresponding figures are as attached. Figure 7 , 8 shown.

[0124] Figure 8Note: The size of the circle represents the data of genes enriched in the corresponding pathway, and the change from green to red represents a gradually decreasing P value.

[0125] The top 20 KEGG metabolic pathways were screened according to the P value, and a bubble chart was drawn based on the P value. The horizontal axis was represented by the number of genes enriched in the pathway, the size of the bubble represented the number of genes enriched in the corresponding pathway, and the depth of the color represented the significance. The significant enrichment information could be observed intuitively.

[0126] 3.2.5.6 Molecular docking results

[0127] According to the analysis in the previous article, the top 10 important targets were selected for semi-flexible docking with compounds with higher affinity values. The binding energy (affinity) is used to indicate the binding quality of the small molecule to the target protein. A binding energy less than 0 means that the small molecule and the target protein can bind freely. The smaller the value, the higher the possibility of binding.

[0128] The docking results show that all small molecules can enter the active center of the target protein. The small molecules with the best docking with each protein are selected for display; Ergotamine forms a hydrogen bond with THR-160 of AKT1, and the length of the hydrogen bond is Ergotamine forms hydrogen bonds with SER-108, CYS-13, and VAL-12 of BNDF, and the lengths of the hydrogen bonds are Ergotamine forms a hydrogen bond with ARG-207 of CASP3, and the length of the hydrogen bond is Ergotamine forms a hydrogen bond with GLN-611 of CTNNB1, and the length of the hydrogen bond is Ergotamine forms a hydrogen bond with ASN-455 of ESR1, and the lengths of the hydrogen bonds are Ergotamine forms a hydrogen bond with ASP-84 of IL10, and the length of the hydrogen bond is Ergotamine forms a hydrogen bond with PHE-25 of INS, and the length of the hydrogen bond is Ergosterol forms a hydrogen bond with GLU-275 of JUN, and the length of the hydrogen bond is Cholesterol forms a hydrogen bond with SER-74 of NFKB1, and the length of the hydrogen bond is Ergotamine forms hydrogen bonds with ARG-103 of TNF, and the lengths of the hydrogen bonds are The docking results are shown in Table 2.

[0129] Table 2 Docking results of core small molecules and core target proteins

[0130]

[0131]

[0132] Fig. 9 Note: Figure A is the interaction pattern of Ergotamine and AKT1, Figure B is the interaction pattern of Ergotamine and BNDF, Figure C is the interaction pattern of Ergotamine and CASP3, Figure D is the interaction pattern of Ergotamine and CTNNB1, Figure E is the interaction pattern of Ergotamine and ESR1, Figure F is the interaction pattern of Ergotamine and IL10, Figure G is the interaction pattern of Ergotamine and INS, Figure H is the interaction pattern of Ergosterol and JUN, Figure I is the interaction pattern of Cholesterol and NFKB1, and Figure J is the interaction pattern of Ergotamine and TNF.

[0133] In summary, Zhilingfang has a synergistic mechanism of multiple components, multiple targets, and multiple pathways in the treatment of CIPN. Through the analysis of Chinese medicine system pharmacology, 27 active ingredients of Zhilingfang were screened out, which acted on 210 target genes at the intersection of nerve damage and drugs, and constructed a complex "drug-ingredient-target" network. Among them, the top ten ingredients such as Ammonium Oxalate and Ergotamine played an important role in the network.

[0134] The core targets include AKT1, TNF, INS, JUN, NFKB1, IL10, CASP3, BDNF, CTNNB1, ESR1, etc., which are involved in a variety of biological processes and signaling pathways. GO enrichment analysis showed that Zhilingfang mainly affects biological functions such as response to exogenous stimuli, signal transduction, inflammatory response, and apoptosis, involving cellular components such as plasma membrane, cytoplasm, and mitochondria, as well as molecular functions such as protein binding and metal ion binding. KEGG pathway enrichment analysis screened out 111 pathways, such as the role of AGE-RAGE signaling pathway in diabetic complications, tumor necrosis factor signaling pathway, and the interaction between neuroactive ligands and receptors.

[0135] The results of molecular docking showed that active ingredients such as Ergotamine formed hydrogen bonds with core targets and had strong binding activity. For example, Ergotamine interacted with targets such as AKT1, BNDF, and CASP3 through hydrogen bonds, which may regulate related signaling pathways and thus play a therapeutic role in CIPN. These results suggest that Zhilingfang may improve the symptoms of CIPN patients by regulating multiple signaling pathways, inhibiting inflammatory responses, regulating cell apoptosis, and promoting the protection and repair of nerve cells, etc., which provides a theoretical basis at the molecular level for the treatment of CIPN with Zhilingfang, and also provides important clues for further studying its mechanism of action and developing new treatment strategies.

Claims

1. The Chinese medicine Zhiling prescription is characterized in that: The invention comprises the following raw materials in parts by weight: 18-22 parts of clematis root, 8-12 parts of mulberry twig, 8-12 parts of cassia twig, 8-12 parts of bombyx batryticatus and 8-12 parts of scorpion.

2. The Chinese medicine Zhiling prescription as claimed in claim 1, characterized in that Also includes 2 to 4 portions of centipedes.

3. The preparation method of the Chinese medicine Zhiling Fang according to claim 2, characterized in that: The steps include: (1) Take the following raw materials in parts by weight: 18-22 parts of Clematis chinensis, 8-12 parts of Morus alba, 8-12 parts of Cinnamomum cassia, 8-12 parts of Bombyx batryticatus, and 8-12 parts of Scorpio, wash, dry and crush; Alternatively, take the following raw materials in parts by weight: 18-22 parts of clematis, 8-12 parts of mulberry branches, 8-12 parts of cassia twigs, 8-12 parts of bombyx batryticatus, 8-12 parts of scorpions, and 2-4 parts of centipedes, wash, dry and crush; (2) Take the crushed raw material in (1), add 5-10 times the weight of the raw material water, boil over high heat, and then simmer over low heat for 10-20 minutes. Filter and take the filtrate. Simmer the remaining residue in the same way again. Filter and take the filtrate. Combine the two filtrates, cool and bag.

4. The preparation method of the Chinese medicine Zhiling Fang according to claim 3, characterized in that: (2), the weight ratio of raw materials to water is 1:5~7.

5. The preparation method of the Chinese medicine Zhiling Fang according to claim 3, characterized in that: The steps include: (1) Take 20 g of Clematis chinensis, 10 g of Morus alba, 10 g of Cinnamomum cassia, 10 g of Bombyx batryticatus, and 10 g of Scorpio, wash them, dry them, and crush them; Alternatively, take 20 g of Clematis chinensis, 10 g of Morus alba, 10 g of Cinnamomum cassia, 10 g of Bombyx batryticatus, 10 g of Scorpio scorpion, and 3 g of Scolopendra subspinipes, wash them, dry them and crush them; (2) Take the crushed raw material in (1), add 400 g of water, bring to a boil over high heat, then simmer over low heat for 10-20 min, filter, take the filtrate, and simmer the remaining residue again in the same way. Filter and take the filtrate, combine the two filtrates, control the filtrate to 90-110 mL, and bag it after cooling.

6. Use of the Chinese medicine Zhiling Fang as described in any one of claims 1 or 2 as a drug for treating peripheral neuropathy.

7. The use according to claim 6, characterized in that The peripheral neuropathy drug is a drug used for peripheral neuropathy induced by chemotherapy drugs.

8. The use according to claim 6, characterized in that The medicine is an oral medicine.

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