Traditional Chinese medicine zhilingsheng formula, preparation method and use thereof

Abstract

The present application relates to the technical field of nerve injury drugs, in particular to a traditional Chinese medicine Zhiling recipe, a preparation method and application thereof in preparation of drugs for relieving peripheral neuropathy, wherein the traditional Chinese medicine Zhiling recipe comprises the following raw materials in parts by weight: Radix Aristolochiae 18-22 parts, Ramulus Mori 8-12 parts, Ramulus Cinnamomi 8-12 parts, Bombyx Batryticatus 8-12 parts, Scorpio 8-12 parts and Solpugid 2-4 parts. The traditional Chinese medicine Zhiling recipe provided by the present application has good effects on peripheral neuropathy, can greatly relieve the numbness of limbs, and through retrospective clinical observation research and mechanism research by network pharmacology, the present application provides medical evidence for treating peripheral neuropathy induced by chemotherapy in traditional Chinese medicine.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of medicine, in particular to Zhiling Decoction, a preparation method and the use of Zhiling Decoction as a drug for treating peripheral neuropathy. BACKGROUND

[0002] Chemotherapy-induced peripheral neuropathy (CIPN) is a common, dose-limiting adverse reaction associated with chemotherapy drugs, about 50%-90% of patients receiving chemotherapy will develop CIPN, of which 30%-40% will develop into chronic adverse reactions. CIPN is clinically common and seriously affects the quality of life of patients and even the normal progress of treatment. Currently, 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), and other drug-related factors include single dose, cumulative dose and duration of drug exposure, other drug interactions in combination regimens, etc. The pathogenesis of CIPN is not fully understood, 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 drug, 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 classics. Its main manifestations are symmetrical paresthesia in the distal extremities, motor nerve dysfunction and autonomic nervous dysfunction. According to the initial characteristics of CIPN, which are mainly paresthesia such as numbness and pain in the fingers and toes, the symptoms are very similar to "Bi" and "Wei" syndromes. Combined with the specific symptoms of CIPN, CIPN can be attributed to "numbness" in traditional Chinese medicine, which belongs to the category of "blood stasis"; over time, muscle atrophy, severe motor nerve dysfunction symptoms such as limb paralysis, etc. When it belongs to the category of "flaccid syndrome" in traditional Chinese medicine.

[0004] There is no satisfactory treatment for CIPN. In modern clinical research, the treatment of CIPN is different, but the treatment principle is always based on "deficiency", "cold" and "stasis". The internal treatment methods for CIPN include GJG, Huangqiguishewu Decoction, Buyanghuangw Decoction, etc. The external treatment methods include moxibustion, fumigation, acupuncture, etc. In addition, the drugs for the treatment of peripheral neuropathic pain are (1) duloxetine (60mg / d), which can be used as the first-line drug for the treatment of neuropathic pain caused by CIPN (evidence level: grade II, recommendation level: strong recommendation); (2) pregabalin (150mg / d), which can be used as the preferred drug for the treatment of peripheral neuropathic pain related to taxane drugs (evidence level: grade II, recommendation level: strong recommendation); (3) as a back-line treatment, CIPN can be treated with non-opioid (non-steroidal anti-inflammatory drugs) and / or opioid analgesics according to clinical experience, and the influence of complications and combined drugs should be paid attention to (evidence level: grade III, recommendation level: strong recommendation); (4) local peripheral neuropathic pain should be treated with local treatment, such as capsaicin patch (179mg), lidocaine patch, other patches and gel preparations, etc. (evidence level: grade II, recommendation level: strong recommendation).

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

[0006] Therefore, the application provides a traditional Chinese medicine branch Ling formula, a preparation method and a use of the traditional Chinese medicine branch Ling formula as a drug for treating peripheral neuropathy.

[0007] The application group summarizes the clinical experience for many years, refers to the treatment experience of traditional Chinese medicine for herpes zoster postherpetic neuralgia, stroke sequelae and other classical prescriptions, and formulates a traditional Chinese medicine composition with good effect for relieving CIPN. Since the traditional Chinese medicine composition contains main active ingredients such as mulberry branch, cassia twig, radix clematidis, cantharis and scorpion, the traditional Chinese medicine composition is named "branch Ling formula" in the application.

[0008] The formula of the traditional Chinese medicine branch Ling formula provided by the application comprises 18-22 parts of radix clematidis, 8-12 parts of mulberry branch, 8-12 parts of cassia twig, 8-12 parts of cantharis, and 8-12 parts of scorpion. Preferably, it further comprises 2-4 parts of cantharis.

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

[0010] (1) 20g of radix clematidis, 10g of mulberry branch, 10g of cassia twig, 10g of cantharis, 10g of scorpion and 3g of cantharis are taken, washed, dried and crushed;

[0011] (2) Take the raw materials in (1), first burn to open with strong fire, then simmer for 10-20 min with weak fire after adding water, filter, take the filtrate, take the filtrate again after the remaining filter residue is simmered once in the same way, filter, and take the filtrate after cooling, and then bag. The medicine prepared by the above method is used in the form of internal administration, and the dosage of Zhiling Decoction is 2 bags per day, 100 mL per bag.

[0012] The application of the traditional Chinese medicine Zhiling Decoction in the preparation of a medicine for treating peripheral neuropathy is also an important protection content of the present application, and the peripheral neuropathy is caused by chemotherapy induction, such as induction by taxane chemotherapy drugs, and of course, the above-mentioned neuropathy can also be caused by other reasons, and Zhiling Decoction can be used as a medicine for treating the above-mentioned neuropathy.

[0013] The formula of the Zhiling Decoction provided by the present application comprises:

[0014] Radix et Rhizoma Clematidis, with a bitter taste and a warm nature, has the effects of expelling wind and dampness and benefiting joints, and is used for treating rheumatic arthralgia, joint pain and numbness, and is the monarch drug.

[0015] Radix et Rhizoma Morindae Officinallis, with a bitter taste and a warm nature, has the effects of expelling wind and dampness and benefiting joints, and is used for treating rheumatic arthralgia, joint pain and numbness.

[0016] Radix et Rhizoma Cinnamomi, with a bitter taste and a warm nature, has the effects of dispelling cold and pain and promoting yang and qi, and is used for treating cold and blood stasis pain, and is the minister drug.

[0017] Cicada, with a salty and bitter taste and a warm nature, has the effects of calming wind and relieving convulsion, expelling wind and relieving pain, and resolving phlegm and eliminating mass, and is used for treating rheumatic arthralgia, joint pain and numbness, and is the assistant drug. The present application has the following beneficial effects:

[0018] (1) The Zhiling Decoction provided by the present application can effectively treat CIPN numbness and has a simple formula, and Radix et Rhizoma Clematidis is used as the monarch drug, Radix et Rhizoma Cinnamomi is used as the minister drug, Cicada is used as the assistant drug, and Scorpio is used as the minister drug, and Radix et Rhizoma Morindae Officinallis and Eupolyphaga are used as the auxiliary components, and the Zhiling Decoction has the effects of dredging meridians and relieving limb numbness.

[0019] (2) The Zhiling Decoction provided by the present application can effectively treat peripheral neuropathy caused by chemotherapy drugs, and can effectively reduce the numbness degree of CIPN and reduce the numbness plane through internal administration, and has no serious adverse reactions.

[0020]

[0021] ​(3) Through the retrospective study, systematic analysis of existing clinical data, evaluation of Ziling prescription in the treatment of chemotherapy-induced peripheral neuropathy, and mechanism research by network pharmacology, so as to provide strong evidence-based medicine and basic research evidence for TCM treatment of chemotherapy-induced peripheral neuropathy. BRIEF DESCRIPTION OF DRAWINGS

[0022] Figure 1 is the technical route of the clinical efficacy and safety verification of Chinese medicine Ziling prescription for treating peripheral neuropathy induced by chemotherapy drugs (CIPN);

[0023] Figure 2 is the 3.2.2 nerve damage related target map in example 3;

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

[0025] Figure 4 is the 3.2.4 core target and network interaction in example 3;

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

[0027] Figure 6 is the 3.2.5.1 GO gene function enrichment analysis diagram of drug-disease intersection genes using DAVID database (II) in example 3;

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

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

[0030] Figure 9 is the 3.2.5.6 molecular docking result diagram in example 3. DETAILED DESCRIPTION

[0031] In order to enable those skilled in the art to better understand the present application, the present application will be further described in conjunction with specific embodiments.

[0032] Example 1

[0033] 1.1 Preparation of Chinese medicine Ziling prescription

[0034] The Chinese medicine Ziling prescription is used in the form of decocting medicine and taking orally. The method of decocting Ziling prescription is as follows:

[0035] (1) Take Wuling San 20g, Sangzhi 10g, Guizhi 10g, Jiangcan 10g, Quanxie 10g, and Centipede 3g;

[0036] (2) Add the raw materials in (1) to 400g of water, and boil under strong fire, then simmer under weak fire for 20 minutes. Filter, take the filtrate, control the filtrate to be about 100mL, then add 300g of water to the remaining filter residue, and boil under strong fire, then simmer under weak fire for 15 minutes. Filter, take the filtrate, control the filtrate to be about 100mL, and combine the two filtrates. Cool and bag.

[0037] Dosage: The dosage of Zhiling Formula is 2 bags per day, 100mL per bag.

[0038] 1.2 For the Zhiling Formula in 1.1, further retrospective observation clinical research needs to be reasonably designed to verify its efficacy and safety.

[0039] The commonly used evaluation methods are comprehensive clinical evaluation tools (adverse event general term standard) and patient-reported outcome measures. The clinical evaluation tools include EORTC CIPN20 scale, and adverse reaction evaluation scales for specific drugs, such as specific scales in the cancer patient quality of life evaluation scale system (taxane and oxaliplatin neurotoxicity special evaluation scale). The above evaluation tools have certain diagnostic value, but there is no evidence to show that any of the evaluation tools is significantly superior.

[0040] The present application systematically analyzes the existing clinical data by carrying out retrospective research, evaluates the efficacy and safety of Zhiling Formula in the treatment of peripheral neuropathy induced by taxane chemotherapy drugs, and thus provides a strong evidence-based medical basis for the treatment of peripheral neuropathy induced by chemotherapy (such as the Figure 1

[0041] The specific verification scheme is as follows:

[0042] 1.2.1 Retrospective observation clinical research scheme

[0043] (1) Research object

[0044] Inclusion criteria: from January 2022 to June 2024, patients who visited the outpatient department or ward of China Medical Science Institute Cancer Hospital; pathologically diagnosed as malignant tumor; received taxane, platinum and other chemotherapy drugs; peripheral nerve damage occurred and was determined as CIPN; age 18-80 years old; have complete medical record data.

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

[0046] (3) Data collection​

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

[0048] Neuropathy-related information: CIPN onset time, symptoms (e.g., numbness, pain, paresthesia, etc.) and severity (EORTC QLQ-CIPN20 scale and numbness plane).

[0049] Zhilings Decoction usage: dosage, course of treatment, time of starting use, etc.

[0050] Pre- and post-treatment related examination results: blood routine, urine routine, stool routine, liver and kidney function, electrocardiogram, etc.

[0051] (4) Efficacy evaluation index

[0052] Primary index: change in score on numbness symptoms in EORTC QLQ-CIPN20 scale (e.g., hand numbness, toe or foot numbness, etc.) before and after treatment.

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

[0054] Statistical analysis method: measurement data is represented by (x ± s), and paired t-test is used to compare the changes in primary index before and after treatment; count data is represented by rate (%), and chi-square test (or Fisher's exact test, if applicable) is used to compare the differences between different groups. P < 0.05 is considered statistically significant.

[0055] 1.2.2 Results of retrospective observational clinical study

[0056] (1) Drug preparation

[0057] Wei Lingxian 20g, Sangzhi 10g, Guizhi 10g, Jiangcan 10g, Quanxie 10g, Wugong 3g per day; decocted in water and taken orally.

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

[0059] (2) Patient's basic information: A total of 32 patients were included, 14 males and 18 females; age range was 34-75, with a median age of 63.5 (49.75, 69.5) years. Tumor types included breast cancer in 8 cases, lung cancer in 5 cases, digestive tract malignancies in 16 cases, and other malignancies in 3 cases. Chemotherapy regimens mainly involved albumin paclitaxel in 17 cases, oxaliplatin in 12 cases, and others in 3 cases.

[0060] Zhilings Decoction usage: Patients took Zhilings Decoction at a dose of one dose per day, taken twice a day, with a course of 14 days, and took 2-4 courses.

[0061] (3) Efficacy evaluation results

[0062] Primary index: The average score of numbness symptom in EORTC QLQ-CIPN20 scale before treatment was [the score before treatment (3.06±0.24)], the average score after treatment was [the score after treatment (2.10±0.52)], and the difference between the scores before and after treatment was [the difference (0.47±0.61)]. The paired t test showed that P<0.001, indicating that the score of numbness symptom after treatment was significantly reduced, i.e. the degree of numbness was obviously reduced.

[0063] Secondary index

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

[0065] <0.05, with statistical difference, suggesting that the pain symptom was relieved.

[0066] Patient self-perceived improvement: 72% of the patients indicated that the symptoms such as numbness and pain were obviously improved, 65% of the patients believed that the quality of life was improved, and 52% of the patients believed that the numbness plane was reduced.

[0067] Safety evaluation results: During the treatment, 1 patient had skin rash, which subsided after anti-allergic treatment. No serious adverse reactions were found, and the blood routine, urine routine, stool routine, liver and kidney function and electrocardiogram examination showed no obvious abnormal changes related to Zhiling Decoction treatment.

[0068] 1.2.3 Retrospective observation of clinical research results analysis

[0069] (1) Effectiveness analysis

[0070] From the primary index EORTC QLQ-CIPN20 scale numbness symptom score, it was significantly reduced after treatment, indicating that Zhiling Decoction had obvious effect on reducing the numbness symptom of CIPN patients. This may be related to the effect of dredging meridians of Weilingxian, Sangzhi and Guizhi in Zhiling Decoction. Weilingxian can directly reach the disease site to improve the numbness symptom, and Sangzhi and Guizhi can assist Weilingxian to strengthen the treatment of numbness and pain, and the monarch and the minister work together to play the effect of dredging meridians and stopping numbness.

[0071] The improvement of secondary indicators such as pain score and quality of life score further supports the effectiveness of Zhiling Decoction. The relief of pain symptoms may be related to the effect of relieving meridian obstruction and relieving pain of drugs such as scorpion and canary grass. Scorpion can relieve wind and dredge collaterals, and canary grass can dispel wind and relieve pain. Both of them assist the monarch drug and enhance the analgesic effect, thereby improving the quality of life of patients. The improvement of neurological function related indicators (if any) suggests that Zhiling Decoction may have a certain repair effect on nerve damage, but further research is needed to confirm. The improvement of patients' self-feeling is also consistent with the changes of objective indicators, indicating that Zhiling Decoction has been subjectively recognized by patients in improving clinical symptoms.

[0072] (2) Safety analysis: During the observation, although a few patients had skin rash adverse reactions, they were relieved after appropriate treatment, and there were no serious adverse reactions, and the related examinations did not suggest obvious damage to the function of the organs. This shows that Zhiling Decoction has good safety in clinical application, and each herb in the drug composition does not show obvious toxic reactions under the conventional dose. However, due to the limitations of retrospective studies, such as incomplete monitoring of adverse reactions, further safety monitoring is still needed.

[0073] The following Table 1 shows the treatment of 32 patients (involving multiple tumor types) treated with Zhiling Decoction for CIPN in the TCM clinic from May 2022 to November 2022.

[0074] Table 1: Patient Statistics (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 shown in Table 1 above, the Zhiling Decoction composed of Weilingxian 20g, Sangzhi 10g, Guizhi 10g, Jiangcan 10g, Quanxie 10g, and Wugong 3g can effectively reduce the numbness degree (upper and lower limbs or upper limbs or lower limbs) of CIPN, reduce the numbness plane, and the longest time to take effect is less than 3 months, and the shortest time to take effect is about one week. It can greatly improve the quality of life of patients and has no serious adverse reactions, providing a more comprehensive treatment plan for CIPN.

[0080] Example 2

[0081] The prescription used in this example is: Wei Lingxian 20g, Sangzhi 10g, Guizhi 10g, Jiangcan 10g, Quanxie 10g. The preparation method and medicinal effect verification of Zhiling Prescription are the same as those in Example 1. The only difference between the prescription in this example and that in Example 1 is that there is no addition of Mantis.

[0082] In the previous clinical observation, it was found that the effect of Wei Lingxian 10g, Sangzhi 10g, Guizhi 10g, Jiangcan 10g, and Quanxie 9g was not obvious for some patients, but after adding 2 pieces of Mantis (about 3g) in the prescription, the symptoms of the patients improved significantly after 14 days. Based on this, it is speculated that in this example, the addition of Mantis may have a certain positive effect on the overall efficacy, and if this flavor is not added, the corresponding efficacy will be reduced. For example, in the experiment on 6 patients of the same gender, same type of tumor, and almost the same cycle, it was found that the average numbness relief time of the patients in Example 2 was longer, and the average numbness relief time reached 21 days compared with Example 1. This may be because Mantis has the effect of searching for wind and dredging collaterals, attacking toxins and resolving knots, and synergizes with other drugs in the prescription to enhance the effect of dredging collaterals and relieving numbness. When Mantis is lacking, this synergistic effect is weakened, resulting in a slower numbness relief speed. However, the prescription in Example 2 may still have an improvement effect on the symptoms of the patients to some extent, but the effect is not as significant as the complete prescription (i.e., the prescription containing Mantis in Example 1), which further illustrates the rationality and synergy of the various drug compositions in Zhiling Prescription.

[0083] Comparative Example 1

[0084] The prescription provided in this comparative example is different from Example 1 in that it only contains Wei Lingxian 20g and Jiangcan 10g. In the previous clinical observation, a patient used Wei Lingxian 10g, Sangzhi 10g, Guizhi 10g, and Jiangcan 10g alone, and the effect was not obvious after 14 days of use. Only after adding Quanxie and other drugs did the patient improve. It can be inferred that the prescription in this comparative example may have insufficient efficacy.

[0085] Experiments were conducted on 6 patients of the same gender, same type of tumor, and almost the same cycle, and the results showed that the average numbness relief time was longer, reaching 28 days (compared with the complete prescription in Example 1, it is assumed that the numbness relief time is significantly prolonged here). Although Wei Lingxian has the effect of dispelling wind and dampness and dredging collaterals, and Jiangcan can dispel wind and relieve pain, resolve phlegm and resolve knots, the lack of synergistic compatibility of other drugs in the prescription, especially the lack of drugs such as Quanxie with strong collateral dredging and pain relieving effects, as well as Sangzhi and Guizhi which assist in collateral dredging, results in a significant reduction in overall efficacy. Only these two drugs cannot fully and effectively play a role in the complex pathogenesis of CIPN, and cannot fully improve the numbness symptoms of patients, resulting in a longer numbness relief time, which further proves the importance of the synergistic effect of multiple drugs in Zhiling Prescription.

[0086] Comparative Example 2

[0087] The difference between the formula provided in this comparative example and Example 1 is that only 20 g of radix dipsaci, 10 g of ramulus cinnamomi, 10 g of cantharis and 10 g of scolopendra are contained, i.e. only monarch, minister and assistant are contained, and no morus alba and scolopendra are added. According to the previous clinical observation, when some key drugs are missing in the formula, the curative effect will be affected. For example, the effect of radix dipsaci 10 g, morus alba 10 g, ramulus cinnamomi 10 g and cantharis 10 g is not obvious for some patients, and it is gradually improved after the addition of scolopendra, but compared with the complete formula (i.e. the formula in Example 1), the effect is still different.

[0088] When experiments were conducted on patients under the same conditions, it was found that the formula had a certain improvement effect on the numbness symptoms of the patients, but compared with Example 1, the average numbness relief time was prolonged to 25 days. Radix dipsaci is the monarch drug, which can dispel wind and dampness and dredge collaterals; ramulus cinnamomi assists radix dipsaci to strengthen the treatment of numbness and pain, and is the minister drug; cantharis assists radix dipsaci to dredge collaterals, and scolopendra searches for wind and dredges collaterals, leading all drugs to the disease site. Although there is a certain synergy among monarch, minister and assistant, the lack of the efficacy of morus alba in dispelling wind and dampness and benefiting joints and the efficacy of scolopendra in searching for wind and dredging collaterals and attacking toxins and resolving knots makes the formula less effective than the complete Zhiling formula in dredging collaterals and relieving numbness. This shows that morus alba and scolopendra, although they are assistant drugs in Zhiling formula, are of great significance to enhancing the overall curative effect and accelerating the relief of numbness symptoms, further embodying the scientificity and integrity of the drug composition of Zhiling formula.

[0089] Comparative Example 3

[0090] The difference between the formula provided in this comparative example and Example 1 is that 10 g of radix dipsaci, 10 g of morus alba, 10 g of ramulus cinnamomi, 10 g of cantharis and 10 g of scolopendra are contained. The previous clinical observation found that the change of the dosage of radix dipsaci may affect the curative effect, such as the effect of radix dipsaci 10 g and other drugs for 14 days is not obvious for some patients, and the symptoms are improved after adjusting the formula.

[0091] For this comparative example, when experiments were 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 case). Radix dipsaci is the monarch drug in Zhiling formula, and the reduction of its dosage may weaken the overall efficacy of the formula in dispelling wind and dampness and dredging collaterals. Although the other drug ingredients are the same as in Example 1, the insufficient dosage of the monarch drug may not be able to fully play a leading role, affecting the treatment effect of the entire formula on CIPN, resulting in a slower numbness relief speed. This further shows that the dosage and ratio of each drug in Zhiling formula are verified by practice, and reasonable drug dosage is crucial to exert the best curative effect.

[0092] Example 3 Network pharmacology research results

[0093] The network pharmacology research was carried out by Example 3 on the medicine obtained in Example 1 (hereinafter referred to as Zhiling Decoction), 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 of Zhiling Decoction and prediction of target points

[0096] The Traditional Chinese Medicine Database and Analysis Platform (TCMSP, https: / / tcmsp-e.com / ) was used to search for all active ingredients of Wilingxian, Jiangcan, Sangzhi, Guizhi, Quanxie and Eupolyphaga with oral bioavailability (OB) ≥ 30% and drug-likeness (DL) ≥ 0.18 as conditions, and active ingredients were screened. For drugs that could not be retrieved from TCMSP and obtained high reliability proteins as candidate genes by logging into the BATMAN-TCM database (http: / / bionet.ncpsb.org.cn / batman-tcm / ). The target points obtained after collation were calibrated by Uniprot (https: / / www.uniprot.org / ), non-human genes were removed, invalid and duplicate target points were deleted, and standardized gene names were obtained.

[0097] 3.1.2 Acquisition of nerve damage related target points

[0098] By inputting the keyword "Nerve damage" in GeneCards (https: / / www.genecards.org / ), OMIM (https: / / omim.org / ) and DisGENET (https: / / www.disgenet.org / ) databases, disease related target points were obtained, all target points in the two databases were integrated in excel, duplicate genes were removed, and Uniprot database was corrected to obtain disease target gene information.

[0099] 3.1.3 Prediction results of drug-disease target points

[0100] The obtained drug component target points and disease target points were mapped to each other, and then Venn diagram was made to obtain intersection genes. Then Cytoscape3.8.2 software was used to construct "drug-component-action target point" network.

[0101] 3.1.4 Construction of target protein interaction network

[0102] To further study the protein-protein interactions of Ziling Fang in the treatment of nerve injury, the drug-intersection genes were uploaded to the interaction database String (https: / / string-db.org / ) for protein-protein interaction network construction (PPI) database; the species was set to "Homo sapiens", the minimum interaction score was set to 0.7 to ensure the reliability of the study, and the rest of the parameters were kept at the default settings. 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 Degree. The larger the node, the larger the Degree value. The thickness of the edge was used to reflect the size of Combine score. The thicker the edge, the larger the Combine score. The core target points were selected to make a protein-protein interaction network diagram.

[0103] 3.1.5 GO enrichment analysis and KEGG pathway analysis

[0104] The drug-disease intersection genes were uploaded to the DAVID database (https: / / david.ncifcrf.gov / summary.jsp), and the gene identifier was selected

[0105] OFFICIAL_GENE_SYMBOL, and the species was set to HomoSapiens. The DAVID6.8 GO gene function was used to annotate the role of the target proteins for treating nerve injury in gene function from three aspects of biological process (Biological Process, BP), cellular component (Cellular Component, CC), and molecular function (Molecular Function, MF). To clarify the target points of Ziling Fang in the treatment of stable nerve injury, KEGG pathway enrichment analysis was performed. The top 10 of BP, CC, and MF in GO function, and the 20 pathways related to stable nerve injury (P<0.01) were selected as the main gene function enrichment processes and signal pathways of Ziling Fang in the treatment of stable nerve injury. The mechanism of Ziling Fang in the treatment of stable nerve injury was predicted.

[0106] 3.1.6 Molecular docking

[0107] ​Generally speaking, the higher the Degree value in a network, the more important its position is. That is to say, the higher the Degree value of a protein in a protein interaction network, the more important role it plays in the treatment of nerve injury. Molecular docking was performed on active ingredients and key targets using AutoDock Vina (1.1.2) to verify their interaction activity. The specific method is as follows: (1) Download the SDF format of the compound from the PubChem database, then import Chembio3D for energy minimization, then import AutodockTools-1.5.6 for hydrogen addition, charge calculation, charge distribution, and setting of rotatable bonds, and save as "pdbqt" format; (2) Download the key target protein (human protein first, original ligand with high structural similarity to the active ingredient to be docked, and high resolution) from the PDB (http: / / www.rcsb.org / ) database; (3) Import PyMoL (2.3.0) to remove the original ligand and water molecules, then import the protein into AutoDocktools (v1.5.6) for hydrogen addition, charge calculation, charge distribution, and atom type designation, and save as "pdbqt" format; (4) Use POCASA 1.1 to predict protein binding sites, and the size of the grid box is set to 60x60x60 (the distance between each grid point is ), and the rest of the parameters are default settings. (5) Use PyMOL2.3.0 for interaction mode analysis.

[0108] 3.2 Research results

[0109] 3.2.1 Active ingredients of Zhiling Decoction and target prediction

[0110] After searching TCMSP, OB≥30%, DL≥0.18, and excluding invalid ingredients, 3 ingredients of Wulingxian, 3 ingredients of Sangzhi, and 6 ingredients of Guizhi were collected. After screening by TCMSP, no ingredients of Jiegeng, Quanxie, and Goujie were included, so through the BATMAN-TCM database, 10 ingredients of Jiegeng, 3 ingredients of Quanxie, and 5 ingredients of Goujie were obtained under the condition of Scorecutoff>20. A total of 27 ingredients were obtained.

[0111] 3.2.2 Nerve injury related targets

[0112] Take Relevance score > 5 as the standard for GeneCards database to include target points, 2766 target points are included, 74 and 484 target points are obtained through OMIM, DisGENET database respectively, after deduplication, 2835 target points related to nerve damage are obtained, and the obtained genes are corrected through Uniprot database. The intersection of the action target points of the drug and the target genes of nerve damage is obtained, and 210 intersection target genes of nerve damage and drugs are obtained as the interactive target genes of drug treatment of nerve damage. (As shown in Figure 2

[0113] 3.2.3 Drug-ingredient-target prediction results

[0114] Using drug-ingredient-target data, construct "network.xlsx" file and "type.xlsx" file, and import the file into Cytoscape3.7.2 for drawing. Topological analysis is performed on the network, and the network has 241 nodes and 372 edges. The top ten ingredients in terms of degree value are Ammonium Oxalate, Ergotamine, kaempferol, Tyrosin, Cholesterol, beta-sitosterol, Lupeol, Ergosterol, Stigmasterol and Leucine.

[0115] Note: rectangles represent target points, ellipses represent drug ingredients, and diamond nodes represent traditional Chinese medicines (as shown in Figure 3

[0116] 3.2.4 Core target points and network interaction

[0117] After taking the intersection of the action target points of all drugs and the target genes of nerve damage, 210 intersection target genes of nerve damage and drugs are obtained, which are the interactive target genes of drug treatment of nerve damage. The obtained 20 intersection target genes are imported into String (https: / / string-db.org / ) database for protein-protein interaction prediction, and the species is set to Homo Sapiens and the confidence is set to 0.7. Save the network file in TSV format, and import the TSV file into Cytoscape3.7.2 software to draw the protein interaction network, and select the target points with degree value > 5 to be included, as shown in Figure 4 The figure includes 104 nodes and 1194 edges. Topological analysis is performed on the network, and the degree value reflects the size and color of the target point, and the combined score value reflects the thickness of the edge, thereby constructing the protein-protein interaction network, as shown in Figure 4 ​​AKT1, TNF, INS, JUN, NFKB1, IL10, CASP3, BDNF, CTNNB1, ESR1 are core target points.

[0118] 3.2.5 Biofunction enrichment analysis

[0119] 3.2.5.1 GO enrichment analysis

[0120] Taking the drug-disease intersection genes, GO gene function enrichment analysis was performed using the DAVID database, and 494 GO entries were screened out. Taking P<0.01 as the standard, 355 main entries of significantly enriched biological functions (BP) of treating nerve damage were screened out, mainly involving response to exogenous stimulus, response to lipopolysaccharide, response to hypoxia, signal transduction, positive regulation of gene expression, inflammatory response, negative regulation of apoptosis process, positive regulation of cell proliferation, response to ethanol, positive regulation of apoptosis process; 69 related to cell components (CC) involved in plasma membrane, cytoplasm, membrane component, cytosol, membrane, mitochondria, plasma membrane component, extracellular region, extracellular space, extracellular exosome; and 70 related to molecular function (MF) involved in protein binding, identical protein binding, metal ion binding, protein homodimerization activity, enzyme binding, zinc ion binding, chromatin binding, heme binding, oxidoreductase activity, macromolecular complex binding. The relevant graph is shown in Figure Figure 5 、 6 .

[0121] 3.2.5.2 KEGG pathway enrichment analysis

[0122] Pathway enrichment analysis was performed using the DAVID database, and 111 pathways of Zhiling Decoction and treatment of nerve damage were screened out according to P<0.01, and the pathways related to nerve damage were screened out, respectively:

[0123] Role of AGE-RAGE signaling pathway in diabetic complications, Lipid and atherosclerosis, Fluid shear stress and atherosclerosis, CAMP signaling pathway, Chemical carcinogenesis-receptor activation, Tumor necrosis factor signaling pathway, Neuroactive ligand-receptor interaction, Pathways in neurodegeneration - multiple diseases, IL-17 signaling pathway, HIF-1 signaling pathway, Dopaminergic synapse, MAPK signaling pathway, Tyrosine metabolism, NF-kappa B signaling pathway, Adipocytokine signaling pathway, Toll-like receptor signaling pathway, Relaxin signaling pathway, Calcium signaling pathway, CGMP-PKG signaling pathway, Neurotrophin signaling pathway. The corresponding graph is shown in Figure Figure 7 、 8 .

[0124] Figure 8Note: The size of the circle represents the data of the enrichment of the gene on the corresponding pathway, from green to red, the P value gradually decreases.

[0125] According to the P value, the top 20 KEGG metabolic pathways are screened, and a bubble chart is drawn according to the P value. The horizontal axis represents the number of genes enriched in the pathway, and the size of the bubble represents the number of genes enriched in the corresponding pathway. The depth of color represents the significance, and the significant enrichment information can be observed intuitively.

[0126] 3.2.5.6 Molecular docking results

[0127] According to the analysis in the foregoing, the top 10 important target points and the compounds with higher degree values are selected for semi-flexible docking. The affinity is used to represent the binding of small molecules and target proteins. The smaller the affinity is, the higher the possibility of binding is.

[0128] The docking results show that the small molecules can enter the active center of the target protein. The best small molecule for each protein is selected for plotting and display; Ergotamine forms a hydrogen bond with AKT1 THR-160, and the length of the hydrogen bond is Ergotamine forms a hydrogen bond with BNDF SER-108, CYS-13, and VAL-12, and the lengths of the hydrogen bonds are Ergotamine forms a hydrogen bond with CASP3 ARG-207, and the length of the hydrogen bond is Ergotamine forms a hydrogen bond with CTNNB1 GLN-611, and the length of the hydrogen bond is Ergotamine forms a hydrogen bond with ESR1 ASN-455, and the lengths of the hydrogen bonds are Ergotamine forms a hydrogen bond with IL10 ASP-84, and the length of the hydrogen bond is Ergotamine forms a hydrogen bond with INS PHE-25, and the length of the hydrogen bond is Ergosterol forms a hydrogen bond with JUN GLU-275, and the length of the hydrogen bond is Cholesterol forms a hydrogen bond with NFKB1 SER-74, and the length of the hydrogen bond is Ergotamine forms a hydrogen bond with TNF ARG-103, 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] Figure 9 Figure A is the interaction mode of Ergotamine and AKT1, Figure B is the interaction mode of Ergotamine and BDNF, Figure C is the interaction mode of Ergotamine and CASP3, Figure D is the interaction mode of Ergotamine and CTNNB1, Figure E is the interaction mode of Ergotamine and ESR1, Figure F is the interaction mode of Ergotamine and IL10, Figure G is the interaction mode of Ergotamine and INS, Figure H is the interaction mode of Ergosterol and JUN, Figure I is the interaction mode of Cholesterol and NFKB1, and Figure J is the interaction mode of Ergotamine and TNF.

[0133] In summary, Zhenling Decoction has a synergistic mechanism of multi-component, multi-target and multi-pathway in treating CIPN. Through systematic pharmacological analysis of traditional Chinese medicine, 27 active components of Zhenling Decoction were screened, which act on 210 intersection target genes of nerve damage and drugs, and a complex "drug-component-action target" network is constructed. Among them, components such as Ammonium Oxalate, Ergotamine, etc. play an important role in the network.

[0134] The core target points include AKT1, TNF, INS, JUN, NFKB1, IL10, CASP3, BDNF, CTNNB1, ESR1, etc., which are involved in multiple biological processes and signal pathways. GO enrichment analysis shows that Zhenling Decoction mainly affects the response to exogenous stimuli, signal transduction, inflammatory response, cell apoptosis and other biological functions, involving cell components such as plasma membrane, cytoplasm, mitochondria, and molecular functions such as protein binding, 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 neuroactive ligand-receptor interaction, etc.

[0135] Molecular docking results showed that active ingredients such as Ergotamine formed hydrogen bonds with core targets, and had strong binding activity. For example, Ergotamine interacted with AKT1, BNDF, CASP3 and other targets through hydrogen bonds, which may regulate related signaling pathways, thereby playing a therapeutic role on CIPN. These results suggest that Zhiling Decoction may regulate multiple signaling pathways, inhibit inflammation, regulate cell apoptosis, promote the protection and repair of nerve cells, and thus improve the symptoms of CIPN patients, providing a molecular level theoretical basis for Zhiling Decoction in the treatment of CIPN, and also providing important clues for further studying its mechanism of action and developing new treatment strategies.

Claims

1. A traditional Chinese medicine, characterized in that, The traditional Chinese medicine is named "Zhilingshang", and is composed of the following raw materials in parts by weight: Wilingxian 18-22 parts, Sangzhi 8-12 parts, Guizhi 8-12 parts, Jiangcan 8-12 parts, Quanxie 8-12 parts, and Wugong 2-4 parts.

2. The preparation method of claim 1, characterized in that, The steps include the following: (1) Take the following raw materials in parts by weight: Wilingxian 18-22 parts, Sangzhi 8-12 parts, Guizhi 8-12 parts, Jiangcan 8-12 parts, Quanxie 8-12 parts, and Wugong 2-4 parts, wash, dry and crush; (2) Take the crushed raw materials in (1), add water in a weight ratio of 5-10 times of the raw materials, first burn to open with a strong fire, then simmer for 10-20 min with a weak fire, filter, take the filtrate, and take the filtrate again after the remaining filter residue is simmered once more in the same way, combine the two filtrates, and pack after cooling.

3. The preparation method of claim 2, characterized in that, In (2), the weight ratio of the raw materials to water is 1:5-7.

4. The preparation method of claim 2, characterized in that, The steps include the following: (1) Take Wilingxian 20 g, Sangzhi 10 g, Guizhi 10 g, Jiangcan 10 g, Quanxie 10 g, and Wugong 3 g, wash, dry and crush; (2) Take the crushed raw materials in (1), add 400 g of water, first burn to open with a strong fire, then simmer for 10-20 min with a weak fire, filter, take the filtrate, and take the filtrate again after the remaining filter residue is simmered once more in the same way, filter after the filtrate, combine the two filtrates, control the filtrate at 90-110 mL, and pack after cooling.

5. The use of the traditional Chinese medicine in any one of claims 1 or 2 in the preparation of a drug for treating peripheral neuropathy.

6. Use according to claim 5, characterized in that, The drug for treating peripheral neuropathy is a drug for peripheral neuropathy induced by chemotherapy drugs.

7. The use according to any one of claims 5 or 6, characterized in that, The drug is an oral drug.

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