Application of traditional Chinese medicine composition in preparation of medicine for preventing and / or treating cytokine storm

Abstract

The traditional Chinese medicine composition is mainly prepared from herba schizonepetae, radix saposhnikoviae, rhizoma seu radix notopterygii, radix angelicae pubescentis, poria cocos, rhizoma chuanxiong, radix platycodonis, radix bupleuri, radix peucedani, fructus aurantii and radix glycyrrhizae, and the new application refers to application of the traditional Chinese medicine composition in preparation of a medicine for preventing and / or treating cytokine storm. Research finds that the traditional Chinese medicine composition can rapidly and effectively restrain release of cytokines, and a safe and effective treatment medicine is provided for preventing and treating cytokine storm.

Description

Technical Field

[0001] The present invention belongs to the technical field of traditional Chinese medicines and relates to a new use of a traditional Chinese medicine composition, and specifically relates to a use of a traditional Chinese medicine composition mainly prepared from notopterygium root, angelica dahurica, Poria cocos, siler, schizonepeta tenuifolia, ligusticum chuanxiong, platycodon grandiflorum, bupleurum root, peucedanum peucedanum, fructus aurantii and liquorice in the preparation of a medicine for preventing and / or treating cytokine storm. Background Art

[0002] Cytokine storm (CS) is a systemic inflammatory response that affects the whole body and can be caused by a variety of factors. Diseases that develop from it include cytokine storm syndrome (CSS) and cytokine release syndrome (CRS). CSS is related to infection and autoimmune diseases, while CRS is related to adverse reactions to tumor immunotherapy.

[0003] Cytokine storm syndrome is a term that covers a variety of immune regulation disorders, including systemic symptoms, systemic inflammation and multi-organ dysfunction, which can lead to multi-organ failure if not treated properly. In 1952, Farquhar and Claireaux first described CS symptoms in familial hemophagocytic lymphohistiocytosis (fHLH). In 1993, Ferrara et al. first proposed the term "cytokine storm" in graft versus host disease (GVHD). The term cytokine storm began to be used to describe severe immune responses that occur during infection.

[0004] Cytokine release syndrome is a severe systemic inflammatory response syndrome caused by the activation of immune cells and the release of a large number of cytokines. It was initially classified as cytokine storm syndrome. In 1990, the CD3 monoclonal antibody OKT3 was used in CIK cell therapy. In 2006, after the injection of the humanized CD28 superagonist TGNI412, patients developed a systemic inflammatory response characterized by the rapid release of proinflammatory cytokines. In recent years, with the development of immunotherapy, it has been found that systemic inflammatory responses can also occur after the use of nivolumab, and similar phenomena have also occurred in the use of CAR-T and other therapies. In 2018, the consensus of the American Society of Stomatology (ASBMT) defined CRS as any hyperphysiological response that leads to the activation or participation of endogenous or infused T cells and / or other immune effector cells after any immunotherapy, which is distinguished from the traditional CSS (Liu Liang, Meng Yuan, Gao Quanli. Research progress on cytokine storm syndrome and cytokine release syndrome [J]. Chinese Journal of Clinical Oncology, 2022(012):049.).

[0005] At the beginning of the cytokine storm, inflammation begins in local tissues and then spreads to the whole body through the circulatory system. Activated T cells and immune effector cells in the cytokine storm release a large number of proinflammatory cytokines and chemokines after lysis to stimulate the activation of macrophages, dendritic cells and endothelial cells. These activated cells release more cytokines in an autocrine or paracrine manner, further stimulating the activation of more immune cells and endothelial cells. This cascade amplification reaction causes an imbalance in the immune network through the lack of negative feedback of immune regulation and the continuous self-amplification of positive feedback, ultimately leading to the initiation of a cytokine storm, manifested as excessive inflammatory response in a single or multiple organs, such as cardiovascular symptoms. Symptoms of the blood system, lung symptoms, acute kidney injury, multiple organ failure and even life-threatening.

[0006] Currently, the main types of therapeutic drugs for cytokine storm are as follows: interleukin blockers, TNF antibodies, peroxisome proliferator-activated receptor agonists (PPARs), sphingosine 1-phosphate receptor 1 (S1PR1) agonists, cyclooxygenase (COX) inhibitors, IFN-γ antibodies, immunoglobulins, chemokine receptor antagonists, acetylcholine and its analogs, angiotensin-converting enzyme inhibitors / angiotensin receptor blockers (ACEI / ARB), endogenous anti-inflammatory substances, adrenocortical hormones, etc.

[0007] However, the above-mentioned therapeutic drugs have certain drawbacks and limitations. For example, conventional anti-inflammatory drugs such as non-steroidal anti-inflammatory drugs and corticosteroids will hinder the body's clearance of pathogenic microorganisms such as viruses, thereby limiting their application in the treatment of cytokine storms. In addition, some specific cytokine blockers (such as tocilizumab, anakinra) also have certain limitations, because specific cytokine blockers mainly reduce the production of a single cytokine, and cytokine storms are systemic inflammation caused by multiple cytokines, and the expression of inflammatory cytokines in different types of cytokine storms is also different. Therefore, the development of a safe and efficient drug that can more effectively treat cytokine storms while reducing side effects is a key issue that needs to be solved urgently. Summary of the invention

[0008] The formula of Jingfang Granules comes from Jingfang Baidu Powder in "Health Prescriptions" by Zhang Shiche in the Ming Dynasty. It is mainly used to dispel wind and cold, relieve the symptoms and eliminate dampness. It is called "the universal cure for plague" and "the magic medicine for colds in all seasons" by doctors of all dynasties. Jingfang Granules is a modern formula preparation made by modern pharmaceutical technology based on the formula of Jingfang Baidu Powder. It is a classic reproduction of the ancient famous formula. The present invention is a further study and development of the marketed product "Jingfang Granules" and the use of its formula.

[0009] The present application seeks to protect the use of a Chinese medicine composition in the preparation of a drug for preventing and / or treating cytokine storm.

[0010] The cytokine storm is a phenomenon in which multiple cytokines are rapidly produced in the body due to infection or non-infectious diseases, and the tissue damage caused by it.

[0011] The traditional Chinese medicine composition is mainly prepared from angelica sinensis, angelica pubescens, Poria cocos, siler, schizonepeta tenuifolia, chuanxiong, platycodon grandiflorum, bupleurum, peucedanum peucedanum, fructus aurantii and liquorice.

[0012] Furthermore, the Chinese medicine composition is mainly prepared from the following raw materials:

[0013] Nepeta 15-90 weight parts Saposhnikovia 15-90 weight parts Notopterygium 15-90 weight parts

[0014] Angelica dahurica 15-90 weight parts Bupleurum 9-75 weight parts Peucedanum 9-75 weight parts

[0015] Chuanxiong 15-90 weight parts Citrus aurantium 9-75 weight parts Poria 15-90 weight parts

[0016] 9-75 parts by weight of Platycodon grandiflorum and 3-30 parts by weight of Licorice.

[0017] Furthermore, the Chinese medicine composition is mainly prepared from the following raw materials:

[0018] 45 weight parts of Schizonepeta tenuifolia 45 weight parts of Saposhnikovia divaricata 45 weight parts of Notopterygium incisum

[0019] 45 weight parts of Angelica dahurica 45 weight parts of Radix Peucedanum 45 weight parts of Radix Bupleuri 45 weight parts

[0020] 45 parts by weight of Chuanxiong 45 parts by weight of Citrus aurantium 45 parts by weight of Poria cocos

[0021] 45 parts by weight of Platycodon grandiflorum and 15 parts by weight of Licorice.

[0022] In one embodiment, the cytokine storm is cytokine storm syndrome or cytokine release syndrome.

[0023] In one embodiment, the cytokine storm is caused by a viral infection, a bacterial infection, cancer, an organ transplant, or cancer treatment.

[0024] In one embodiment, the viral infection is caused by one or more of coronavirus, influenza virus, Epstein-Barr virus, respiratory syncytial virus, dengue virus, Zika virus, West Nile virus, smallpox virus, cytomegalovirus and HIV virus.

[0025] In one embodiment, the bacterial infection is caused by Streptococcus pyogenes, Staphylococcus aureus, Klebsiella pneumoniae, or Yersinia pestis.

[0026] In one embodiment, the cancer is a malignant tumor of the nasal cavity and paranasal sinuses, nasopharyngeal cancer, oral cancer, laryngeal cancer, salivary gland tumor, intracranial tumor, thyroid cancer or tongue cancer, lung cancer, esophageal cancer, cardia cancer, breast cancer, or mediastinal tumor, stomach cancer, colorectal cancer and sigmoid colon and rectal cancer, liver cancer, pancreatic cancer and periampullary cancer, biliary tract tumor or small intestine malignant tumor, kidney cancer, bladder cancer, testicular malignant tumor, penis cancer, cervical cancer, endometrial cancer or ovarian cancer, lymphoma, acute leukemia, chronic leukemia, polycythemia, thrombocythemia, myeloma.

[0027] In one embodiment, the cancer treatment comprises one or more of chemotherapy, radiation therapy, and immunotherapy.

[0028] Furthermore, the immunotherapy includes antibody therapy and chimeric antigen receptor T cell therapy.

[0029] Furthermore, the antibody therapy includes monoclonal antibody or bispecific antibody therapy.

[0030] In one embodiment according to any of the foregoing embodiments, the traditional Chinese medicine composition can be added with conventional pharmaceutical excipients to prepare a clinically acceptable preparation.

[0031] Furthermore, the preparations include but are not limited to oral liquids, suspensions, syrups, mixtures, tablets, pills, powders, granules, and capsules.

[0032] The present invention has achieved unexpected beneficial effects:

[0033] Animal experiments have shown that the Chinese medicine composition provided by the present invention can quickly curb the increase in the concentration of IL-6, IL-8, TNF-α, IL-1β, IL-12p70, IL-17, IFN-β, IFN-γ, GM-CSF, CCL2, and CXCL10 cytokines in the plasma of cytokine storm model animals, increase the level of IL-10, and systematically and comprehensively inhibit the overreaction of cytokines, thereby relieving the cytokine storm that has occurred and providing a fast-acting, effective, and safe therapeutic drug for the treatment of cytokine storm. The Chinese medicine composition of the present invention can also enhance the anti-inflammatory effect of hydrocortisone and reduce the white blood cell count to normal levels in a relatively short period of time. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] Figure 1 The serum cytokines IL-6, IL-8, TNF-α, and GM-CSF levels of the animals in each group before administration and 3h, 6h, and 24h after administration. Compared with the model group, *p<0.05, **p<0.01; compared with the hydrocortisone group, &p<0.05, &&p<0.01; compared with the Jingfang granule group, #p<0.05, ##p<0.01.

[0035] Figure 2 The serum cytokines IL-1β, IL-10, IFN-β, and IFN-γ levels of each group of animals before administration and 3h, 6h, and 24h after administration. Compared with the model group, *p<0.05, **p<0.01; compared with the hydrocortisone group, &p<0.05, &&p<0.01; compared with the Jingfang granule group, #p<0.05, ##p<0.01.

[0036] Figure 3 The serum cytokines IL-12p70, IL-17, CCL2, and CXCL10 levels of the animals in each group before administration and 3h, 6h, and 24h after administration. Compared with the model group, *p<0.05, **p<0.01; compared with the hydrocortisone group, &p<0.05, &&p<0.01; compared with the Jingfang granule group, #p<0.05, ##p<0.01.

[0037] Figure 4 and Figure 5 The blood routine test results of each group of animals before administration and 3h, 6h and 24h after administration. Compared with the model group, *p<0.05, **p<0.01.

[0038] Efficacy Example

[0039] F182112 is a bispecific antibody developed and produced by Shandong New Era Pharmaceutical Co., Ltd. that simultaneously targets BCMA and CD3 receptors on the surface of T cells. It can enrich CD3-positive T cells near myeloma cells expressing BCMA, thereby inducing T cells to kill tumor cells. The patent with authorization announcement number CN111138542B discloses F182112 and its construction method. In order to verify the efficacy of the Chinese medicine composition of the present invention in preventing and treating cytokine storm, the inventor established a cytokine storm model by a single intravenous injection of F182112 in crab-eating monkeys. The following examples are used to illustrate the technical scheme of the present invention rather than to limit the scope of protection of the present invention. Those of ordinary skill in the art who modify or replace the technical scheme of the present invention without departing from the essence and scope of the technical scheme of the present invention still belong to the scope of protection of the present invention.

[0040] 1. Experimental Animals:

[0041] Thirty-two male cynomolgus monkeys, weighing 3 to 5 kg, were purchased from Guangzhou Conghua Huazhen Animal Breeding Farm, with production license number: SCXK (Yue) 2015-0028.

[0042] 2. Medication

[0043] Jingfang granules, provided by Shandong New Era Pharmaceutical Co., Ltd.;

[0044] F182112, provided by Shandong New Era Pharmaceutical Co., Ltd.;

[0045] Hydrocortisone injection was provided by Huazhong Pharmaceutical Co., Ltd.

[0046] 3. Experimental Procedure

[0047] 3.1 Animal grouping, modeling and drug administration

[0048] The cynomolgus monkeys were randomly divided into 4 groups according to their body weight, namely, model group, Jingfang granule group, hydrocortisone group, and combination medication group, with 8 monkeys in each group.

[0049] At 8:00 am, Jingfang granules were administered orally to the monkeys in the combination group at a dose of 1.7 g / kg, 10 ml / kg per monkey. The model group was given the same volume of purified water. The monkeys in the hydrocortisone group and the combination group were intramuscularly injected with hydrocortisone injection at a dose of 1.5 mg / kg, with a volume of 0.3 ml / kg.

[0050] At 9:00 am, the model group, Jingfang granule group, hydrocortisone group, and combined medication group were all injected with F182112 in the hind limbs of the animals using a syringe pump. The dosage was 0.5 mg / kg, and the volume of the modeling agent was 5 ml / kg. Each monkey was administered for 5 minutes. According to the results of the long-term toxicity report of F182113, at a dose of 0.5 mg / kg, cytokines in crab-eating macaques will increase rapidly 2-6 hours after administration of F182112.

[0051] At 14:00 in the afternoon, Jingfang granules were administered again by gavage to the monkeys in the combination group at a dose of 1.7 g / kg, 10 ml / kg per monkey. The model group was given the same volume of purified water. At 14:00 in the afternoon, hydrocortisone injection was injected intramuscularly to the monkeys in the hydrocortisone group and the combination group at a dose of 1.5 mg / kg, with a volume of 0.3 ml / kg.

[0052] The dosage and administration method of each group are shown in Table 1.

[0053] Table 1 Animal groups and dosage, concentration, volume and method of administration in each group

[0054]

[0055] 3.2 Detection indicators

[0056] Blood samples were collected one day before model establishment (marked as 0h) and 3h, 6h, and 24h after administration of F182112 to detect blood routine and serum cytokine levels.

[0057] 3.3 Statistical analysis

[0058] SPSS 22.0 software was used to analyze the data. The differences were statistically significant when p < 0.05.

[0059] 4. Results and Conclusions

[0060] 4.1 Changes in serum cytokine levels in each group of animals

[0061] IL-6, IL-8, TNF-α, IL-1β, IL-12p70, and IL-17 are proinflammatory cytokines, among which IL-6 is an important cytokine involved in cytokine storms. The above-mentioned proinflammatory cytokines can stimulate the activation of macrophages, dendritic cells, and endothelial cells. These activated cells release more cytokines in an autocrine or paracrine manner, further stimulating the activation of more immune cells and endothelial cells. IFN-β and IFN-γ are also proinflammatory cytokines, which can help activate dendritic cells, proinflammatory macrophages, neutrophils, effector T cells, B cells, etc., and promote the occurrence and development of inflammatory responses. GM-CSF increases the number of macrophages that produce cytokines at the site of inflammation, causing the inflammatory response to continue and leading to a cascade amplification effect of inflammation. IL-10 is an anti-inflammatory factor that inhibits the release of inflammatory mediators by mononuclear macrophages, thereby inhibiting the secretion of TNF-α, IL-1β, IL-6, IL-8, G-CSF, and GM-CSF caused by LPS and IFN-γ. In addition, it enhances the release of anti-inflammatory factors, such as IL-1 receptor antagonists and soluble TNF-α receptors. CCL2 and CXCL10 are chemokines that control the tropism and recruitment of leukocytes. Recruitment and chemotaxis of leukocytes to the site of inflammation.

[0062] like Figure 1 , Figure 2 and Figure 3 As shown in the figure, before intravenous injection of F182112, the cytokines of animals in each group were at normal levels. The serum cytokine levels of cynomolgus monkeys in each group were detected 3 hours after intravenous injection of F182112. It was found that the levels of cytokines such as IL-6, IL-8, TNF-α, IL-1β, IL-12p70, IL-17, IFN-β, IFN-γ, GM-CSF, CCL2, and CXCL10 in the serum of cynomolgus monkeys in the model group increased rapidly, while the cytokine levels of cynomolgus monkeys in the Jingfang granule group, hydrocortisone group, and combined medication group were lower than those in the model group, and there were significant differences compared with the model group. The level of anti-inflammatory factor IL-10 in the serum of cynomolgus monkeys in the model group increased 3 hours after intravenous injection of F182112, but the increase in IL-10 was smaller than that in other groups. Each group continued to receive medication, and the serum cytokine levels were monitored 6 hours after modeling. It was found that the levels of IL-6, IL-8, TNF-α, IL-1β, IL-12p70, IL-17, IFN-β, IFN-γ, GM-CSF, CCL2, and CXCL10 in the serum of the cynomolgus monkeys in the Jingfang granule group, the hydrocortisone group, and the combined medication group continued to decrease significantly, while the cytokines in the cynomolgus monkeys in the model group were still at a high level, which was significantly different from the model group. The serum IL-10 level of the cynomolgus monkeys in the Jingfang granule group, the hydrocortisone group, and the combined medication group remained at a high level, which was significantly different from the model group.

[0063] Jingfang Granules can quickly inhibit the increased concentrations of IL-6, IL-8, TNF-α, IL-1β, IL-12p70, IL-17, IFN-β, IFN-γ, GM-CSF, CCL2, and CXCL10 cytokines in the plasma of crab-eating monkeys in the cytokine storm model within 3-6 hours, increase the level of anti-inflammatory factor IL-10, and systematically and comprehensively inhibit the excessive response of cytokines, thereby relieving the cytokine storm that has already occurred.

[0064] 4.2 Results of routine blood tests of animals in each group

[0065] Depend on Figure 4 and Figure 5 It can be seen that before intravenous injection of F182112, the white blood cell counts of cynomolgus monkeys in each group were at normal levels. Blood was collected from each group of cynomolgus monkeys 3-6 hours after intravenous injection of F182112 to observe the white blood cell counts. It was found that the white blood cell counts of cynomolgus monkeys in the model group increased rapidly. 3-6 hours after modeling, the white blood cell counts of cynomolgus monkeys in the hydrocortisone group and the combined medication group dropped to the level of normal animals, and there was a significant difference compared with the model group (p<0.01). The white blood cell counts of cynomolgus monkeys in the Jingfang granule group were also lower than those in the model group, but there was no significant difference compared with the model group. It can be seen that Jingfang granules can enhance the anti-inflammatory effect of hydrocortisone.

Claims

1. Use of a traditional Chinese medicine composition in preparing a drug for preventing and / or treating cytokine storm, wherein the traditional Chinese medicine composition is mainly prepared from Notopterygium wilfordii, Angelica dahurica, Poria cocos, Saposhnikovia divaricata, Nepeta tenuifolia, Ligusticum chuanxiong, Platycodon grandiflorum, Bupleurum chinense, Peucedanum peucedanum, Fructus aurantii, and Licorice; the cytokine storm is a phenomenon in which a variety of cytokines are rapidly and massively produced in the body due to infection or non-infection diseases, and the tissue damage caused thereby.

2. The use according to claim 1, characterized in that The Chinese medicine composition is mainly prepared from the following raw materials: Nepeta 15-90 weight parts Saposhnikovia 15-90 weight parts Notopterygium 15-90 weight parts Angelica dahurica 15-90 weight parts Bupleurum 9-75 weight parts Peucedanum 9-75 weight parts Chuanxiong 15-90 weight parts Citrus aurantium 9-75 weight parts Poria 15-90 weight parts 9-75 parts by weight of Platycodon grandiflorum; 3-30 parts by weight of Licorice; Preferably, the Chinese medicine composition is mainly prepared from the following raw materials: 45 weight parts of Schizonepeta tenuifolia 45 weight parts of Saposhnikovia divaricata 45 weight parts of Notopterygium incisum 45 weight parts of Angelica dahurica 45 weight parts of Radix Peucedanum 45 weight parts of Radix Bupleuri 45 weight parts 45 parts by weight of Chuanxiong 45 parts by weight of Citrus aurantium 45 parts by weight of Poria cocos 45 parts by weight of Platycodon grandiflorum and 15 parts by weight of Licorice.

3. The use according to claim 1, characterized in that The cytokine storm is cytokine storm syndrome or cytokine release syndrome.

4. The use according to claim 1, characterized in that The cytokine storm is caused by viral infection, bacterial infection, cancer treatment, organ transplantation or cancer.

5. The use according to claim 4, characterized in that The viral infection is caused by one or more of coronavirus, influenza virus, Epstein-Barr virus, respiratory syncytial virus, dengue virus, Zika virus, West Nile virus, smallpox virus, cytomegalovirus and HIV virus.

6. The use according to claim 4, characterized in that The bacterial infection is caused by Streptococcus pyogenes, Staphylococcus aureus, Klebsiella pneumoniae or Yersinia pestis.

7. The use according to claim 4, characterized in that The cancer treatment includes one or more of chemotherapy, radiotherapy and immunotherapy; the immunotherapy includes antibody therapy or chimeric antigen receptor T cell therapy.

8. The use according to claim 7, characterized in that The antibody therapy includes monoclonal antibody or bispecific antibody therapy.

9. The use according to claim 4, characterized in that The cancers are malignant tumors of the nasal cavity and paranasal sinuses, nasopharyngeal cancer, oral cancer, laryngeal cancer, salivary gland tumors, intracranial tumors, thyroid cancer or tongue cancer, lung cancer, esophageal cancer, cardia cancer, breast cancer, or mediastinal tumors, gastric cancer, colorectal cancer and sigmoid colon and rectal cancer, liver cancer, pancreatic cancer and periampullary cancer, biliary tract tumors or small intestinal malignancies, kidney cancer, bladder cancer, testicular malignancies, penile cancer, cervical cancer, endometrial cancer or ovarian cancer, lymphoma, acute leukemia, chronic leukemia, polycythemia, thrombocythemia, and myeloma.

10. The use according to any one of claims 1 to 9, characterized in that The Chinese medicine composition can be added with conventional pharmaceutical excipients to prepare a clinically acceptable preparation. Preferably, the preparation is selected from oral liquid, suspension, syrup, mixture, tablet, pill, powder, granule, capsule.

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