New uses of il-6 receptor antibodies

Abstract

The application provides use of an IL-6 receptor antibody (for example, tocilizumab) in preparation of a drug for treating a novel coronavirus. The application also provides a pharmaceutical composition for treating a novel coronavirus, which contains an IL-6 receptor antibody (for example, tocilizumab) and a pharmaceutically acceptable carrier or excipient. The IL-6 receptor antibody can improve the oxygenation index, significantly reduce the body temperature of a subject, and has a good effect on treating a novel coronavirus.

Description

Technical Field

[0001] This application is a divisional application of Chinese application number 202010257944.2, filed on April 2, 2020, entitled "A Novel Use of IL-6 Receptor Antibody".

[0002] This invention belongs to the field of biomedicine and relates to the application of IL-6 receptor antibodies in the preparation of drugs for the treatment of novel coronavirus. Background Technology

[0003] 1. Introduction to the novel coronavirus:

[0004] Researchers, through whole-genome sequencing and phylogenetic analysis, have shown that 2019-nCoV is distinctly different from the beta coronaviruses associated with human severe acute respiratory syndrome (SARS) and Middle East respiratory syndrome (MERS). It is a novel coronavirus belonging to the coronavirus family and classified as a beta coronavirus 2b lineage. Furthermore, they found that 2019-nCoV shares 96% homology with bat coronaviruses, thus speculating that bats are its primary origin (Zhou, P. et al, A pneumonia outbreak associated with a new coronavirus of probable bat origin, NatureOnline 3 February 2020). On February 11, 2020, the International Committee on Taxonomy of Viruses (ICTV) announced that the official classification name of the novel coronavirus (2019-nCoV) is severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). On the same day, the World Health Organization (WHO) announced that the official name of the disease caused by this virus is "COVID-19".

[0005] 2. Patients with novel coronavirus exhibit features related to cytokine storm.

[0006] Cytokine storm syndrome (CSS) is attracting increasing attention. The concept of CSS was first proposed by Farquhar and Claireaux in 1952 in familial hemophagocytic lymphohistiocytosis (fHLH). For the next fifty years, there was not much research on it. CSS causes multiple organ dysfunction (MOF), which leads to an extremely high mortality rate, and has once again attracted widespread attention (Randy Q. Cron • Edward M. Behrens et al, Cytokine Storm Syndrome, ISBN 978-3-030-22094-5 (eBook)).

[0007] Cytokine storm (CSS) is characterized by an uncontrolled and dysfunctional immune response involving the sustained activation and proliferation of lymphocytes and monocytes, which secrete large amounts of inflammatory factors. Further comparisons between ICU and non-ICU patients revealed higher plasma concentrations of IL2, IL7, IL10, GCSF, IP10, MCP1, MIP1A, and TNFα in ICU patients, suggesting a correlation between cytokine storm and disease severity. Clinical symptoms in COVID-19 patients also exhibit cytokine storm-related features, including fever, liver dysfunction, lymphopenia, coagulopathy, and nephrotoxicity, and predict poor clinical outcomes. Finding effective treatments to control cytokine storm and prevent progression to severe illness is a crucial strategy for reducing the incidence of severe and critical illness and lowering mortality.

[0008] 3. Interleukin-6 (IL-6) is a key node in cytokine storm syndrome (CSS).

[0009] Interleukin-6 (IL-6) has become a key node in certain cytokine storm syndromes (CSS). IL-6 is a classic multifunctional cytokine involved in inflammatory and febrile responses, exhibiting hormone-like properties and influencing homeostasis. Dysregulation of IL-6 production plays an important pathological role in various inflammatory diseases. IL-6 receptors exist in two forms: membrane-bound IL-6 receptor (mIL-6R) and soluble IL-6 receptor (sIL-6R). IL-6 binds to sIL-6R to form a complex, which then binds to gp130 on the cell membrane, completing trans-signal transduction and exerting a pro-inflammatory effect (Simon A. Jones, Jürgen Scheller, and Stefan Rose-John. Therapeutic strategies for the clinical blockade of IL-6 / gp130 signaling. The Journal of Clinical Investigation (2011)).

[0010] 4. Tocilizumab, an IL-6 receptor antibody, can block IL-6R-mediated signal transduction and clinical overview.

[0011] Tocilizumab is a recombinant humanized anti-human IL-6 receptor monoclonal antibody of the immunoglobulin IgG1 subtype. It can specifically bind to sIL-6R and mIL-6R and inhibit sIL-6R-mediated signal transduction. Roche and Chugai have completed clinical efficacy and safety studies of intravenous tocilizumab in multiple disease areas, including systemic juvenile idiopathic arthritis, adult-onset arthritis, and polyarticular juvenile idiopathic arthritis.

[0012] Intravenous tocilizumab at 8 mg / kg has been approved for rheumatoid arthritis (RA) in more than 70 countries, including Japan and Europe. In the United States, intravenous tocilizumab at 4 mg / kg and 8 mg / kg has been approved for RA patients who have not responded well to anti-TNF agents. Furthermore, tocilizumab has been approved in India and Japan for giant lymph node hyperplasia.

[0013] On April 15, 2011, the U.S. Food and Drug Administration (FDA) approved tocilizumab, alone or in combination with aminopurine, for the treatment of active systemic juvenile idiopathic arthritis (sJIA) (TCZACTEMRA's US Package Insert (USPI), April 2011). On August 1, 2011, tocilizumab was approved in Europe for the treatment of sJIA unresponsive to NSAIDs and systemic corticosteroids. Tocilizumab can be used alone or in combination with methotrexate (MTX) in patients 2 years of age and older (RoACTEMRA Product Features Summary, Roche Resistration Limited 6 Falcon Way Shire Park, Welwyn Garden City, AL71TW, UK, June 4, 2010). The approved dosage is 12 mg / kg intravenously every two weeks for patients weighing less than 30 kg and 8 mg / kg intravenously every two weeks for patients weighing 30 kg or more.

[0014] Based on the Phase 3 clinical trial MRA318JP conducted in Japan, tocilizumab was approved in Japan for the treatment of polyarticular juvenile idiopathic arthritis (pcJIA). Additionally, the Phase 3 study WA19977 examined the efficacy, safety, pharmacokinetic (PK), and disease progression (PD) of tocilizumab in children aged 2–17 years with pcJIA. Summary of the Invention

[0015] This invention discloses for the first time that IL-6 receptor antibodies (such as tocilizumab) can be used to treat the novel coronavirus. Specifically, this invention provides the following technical solutions:

[0016] 1. Use of an IL-6 receptor antibody, preferably tocilizumab (preferably 350-450 mg, more preferably tocilizumab with a single drug dose unit of 350-450 mg) in the preparation of a medicament or kit for treating patients with novel coronavirus, preferably, the patients being mild, moderate, severe or critically ill patients.

[0017] 2. According to the use described in 1 above, the drug or kit further comprises an antiviral agent and optionally a traditional Chinese medicine preparation, wherein when the IL-6 receptor antibody is present in the kit, the IL-6 receptor antibody is administered simultaneously, sequentially, or separately with the antiviral agent and optionally the traditional Chinese medicine preparation, wherein the traditional Chinese medicine preparation includes, but is not limited to, Xiyanping injection, Xuebijing injection, Shenfu injection, and / or Shengmai injection.

[0018] 3. According to the use described in 2 above, wherein the antiviral agent is α-interferon (preferably α-interferon with a single drug dose unit of 5 million U), lopinavir / ritonavir (preferably lopinavir with a single drug dose unit of 200 mg, and ritonavir with a single drug dose unit of 50 mg) or ribavirin (preferably ribavirin with a single drug dose unit of 500 mg); wherein

[0019] Preferably, the single dose of α-interferon is 5 million U twice daily; the single doses of lopinavir / ritonavir are 200 mg / 50 mg per tablet twice daily; and the single dose of ribavirin is 500 mg / dose, 2-3 times daily.

[0020] 4. The use according to any of the preceding claims, wherein the IL-6 receptor antibody is tocilizumab.

[0021] 5. According to the use described in 4 above, wherein the single dose of the tocilizumab is 350 mg, 360 mg, 370 mg, 380 mg, 390 mg, 400 mg, 410 mg, 420 mg, 430 mg, 440 mg or 450 mg, more preferably 400 mg; preferably, the tocilizumab is in a form suitable for intravenous infusion.

[0022] 6. A pharmaceutical composition or single-dose unit (preferably for treating patients with novel coronavirus) containing an IL-6 receptor antibody (preferably 350-450 mg, more preferably 350 mg, 360 mg, 370 mg, 380 mg, 390 mg, 400 mg, 410 mg, 420 mg, 430 mg, 440 mg or 450 mg, even more preferably 400 mg), preferably, the patient being a mild, moderate, severe or critical patient.

[0023] 7. The pharmaceutical composition or single-dose unit according to claim 6 above further comprises an antiviral agent and optionally a glucocorticoid.

[0024] 8. The pharmaceutical composition or single-dose unit according to 7 above, wherein the antiviral agent is α-interferon, lopinavir / ritonavir or ribavirin.

[0025] 9. The pharmaceutical composition or single-dose unit according to any one of 6-8 above, wherein the IL-6 receptor antibody is tocilizumab.

[0026] 10. The pharmaceutical composition or single-dose unit according to 6 above may also be used in combination with traditional Chinese medicine preparations, wherein the traditional Chinese medicine preparations include, but are not limited to, Xiyanping injection, Xuebijing injection, Shenfu injection and / or Shengmai injection.

[0027] 11. An article of an intravenous administration device comprising an IL-6 receptor antibody, wherein a fixed dose of an IL-6 receptor antibody (preferably tocilizumab) is administered to a patient with COVID-19, said fixed dose being 350-450 mg, preferably 350 mg, 360 mg, 370 mg, 380 mg, 390 mg, 400 mg, 410 mg, 420 mg, 430 mg, 440 mg or 450 mg, more preferably 400 mg.

[0028] 12. A method for treating a patient with the novel coronavirus, characterized in that the patient is given 350 mg to 450 mg of tocilizumab, preferably 400 mg of tocilizumab.

[0029] 13. The method of treating patients with novel coronavirus as described in Item 12, wherein the tocilizumab is used in combination with an antiviral drug (such as alpha interferon, lopinavir / ritonavir or ribavirin) and / or an anti-inflammatory drug (such as methylprednisolone).

[0030] In this invention, the subjects selected are patients diagnosed with the novel coronavirus, including but not limited to mild, moderate, severe or critical patients; and the subjects' serum interleukin-6 (IL-6) level is higher than the normal reference range (reference range is <7, unit pg / ml).

[0031] In this invention, the treatment regimen for subjects strictly follows the standard treatment guidelines of the "Diagnosis and Treatment Protocol for Novel Coronavirus (Fifth Edition or Updated)". In addition to standard treatment, subjects are given tocilizumab at a dose of 350-450 mg, preferably 350 mg, 360 mg, 370 mg, 380 mg, 390 mg, 400 mg, 410 mg, 420 mg, 430 mg, 440 mg, or 450 mg, with a more preferred dose of 400 mg, administered over 1 hour. If fever persists within 12 hours, an additional dose is administered (as before), with a maximum of two doses in total. Treatment is discontinued once the body temperature returns to normal.

[0032] Terminology of this invention

[0033] In this invention, the IL-6 receptor antibody is a commercially available monoclonal antibody. Specifically, the IL-6 receptor antibody is commercially available tocilizumab (e.g., purchased from Roche).

[0034] In this invention, the term "single drug dose unit" refers to a single-dose drug formulation containing an IL-6 receptor antibody (especially tocilizumab) to be administered to a patient at the time of the dosing regimen, such as an ampoule, which is a small glass container for holding a drug solution, typically with a capacity of 1 to 25 ml, and is commonly used to store injectable drug solutions.

[0035] In this invention, the clinical manifestations of the novel coronavirus are mainly fever, fatigue, and dry cough. A few patients also experience nasal congestion, runny nose, sore throat, and diarrhea. Severe patients often develop dyspnea and / or hypoxemia one week after the onset of illness, and in severe cases, it rapidly progresses to acute respiratory distress syndrome, septic shock, uncorrectable metabolic acidosis, and coagulation dysfunction. It is worth noting that severe and critically ill patients may have low-grade fever or even no obvious fever during the course of the disease. Mild patients only present with low-grade fever and mild fatigue, without pneumonia.

[0036] In laboratory tests, early-stage COVID-19 patients show normal or decreased total peripheral blood leukocyte count and decreased lymphocyte count. Some patients may exhibit elevated liver enzymes, lactate dehydrogenase (LDH), muscle enzymes, and myoglobin; elevated troponin levels are observed in some critically ill patients. Most patients show elevated C-reactive protein (CRP) and erythrocyte sedimentation rate (ESR), while procalcitonin is normal. In severe cases, D-dimer levels are elevated, and peripheral blood lymphocytes progressively decrease. COVID-19 nucleic acid can be detected in nasopharyngeal swabs, sputum, lower respiratory tract secretions, blood, and stool samples.

[0037] Early chest imaging of the novel coronavirus shows multiple small patchy shadows and interstitial changes, more pronounced in the peripheral lung fields. It then progresses to multiple ground-glass opacities and infiltrates in both lungs; in severe cases, pulmonary consolidation may occur. Pleural effusion is rare.

[0038] The clinical classification of the novel coronavirus can be divided into:

[0039] (1) Mild type. Clinical symptoms are mild and no pneumonia is seen on imaging.

[0040] (2) Common type. Symptoms include fever and respiratory symptoms, and imaging shows signs of pneumonia.

[0041] (3) Severe. Meets any of the following criteria: respiratory distress, M>30 breaths / min; oxygen saturation ≤93% at rest; arterial partial pressure of oxygen (PaO2) / oxygen concentration (FiO2) ≤300mmHg (1mmHg=0.133kPa).

[0042] (4) Critically ill. Patients meeting one of the following criteria: respiratory failure requiring mechanical ventilation; shock; or other organ failure requiring ICU monitoring and treatment.

[0043] In this invention, the term "treatment" includes treating a disease, and generally also includes slowing or reversing the progression of a disease, preventing or slowing the onset of one or more symptoms associated with the disease, reducing and / or alleviating one or more symptoms associated with the disease, reducing the severity and / or duration of the disease and / or any symptoms associated with it and / or preventing further increases in the severity of the disease and / or any symptoms associated with it, preventing, reducing or reversing any physiological damage caused by the disease, and any pharmacological effects that are generally beneficial to the patient being treated.

[0044] Technical effects of the present invention:

[0045] This invention confirms that high levels of inflammatory factors, including IL-6, GM-CSF, and IFN-γ, can be produced in patients with the novel coronavirus.

[0046] In this invention, by administering a treatment regimen of "conventional treatment + tocilizumab" to subjects, it was demonstrated that interleukin-6 (IL-6) receptor antibodies (such as tocilizumab), especially specific doses of tocilizumab, can block the inflammatory response induced by IL-6 in patients with novel coronavirus, specifically manifested as one or more of the following improvements: a significant decrease in C-reactive protein; a significant reduction in fever response, with body temperature returning to normal; an increase in the number of immune cells, specifically manifested as a significant increase in the number of peripheral leukocytes and lymphocytes, which is beneficial to the antiviral response; and a reduction in lung damage, specifically manifested as an improvement in the oxygenation index. Attached Figure Description

[0047] Figure 1 : Displays CD4 counts in patients infected with the novel coronavirus + T cells can produce high proportions of IL-6, GM-CSF, and IFN-γ;

[0048] Figure 2 This study showed that peripheral blood mononuclear cells from patients infected with the novel coronavirus highly express IL-6.

[0049] Figure 3 This shows a comparison of body temperature in patients infected with the novel coronavirus before and after receiving tocilizumab treatment.

[0050] Figure 4 : Displays CT images of patients with novel coronavirus before and after tocilizumab treatment.

[0051] Figure 5Figures 1 and 2 show chest CT scans, demonstrating significant bilateral lung remission following tocilizumab treatment. (AC) Computed tomography (CT) before tocilizumab treatment showed patchy and ground-glass opacities. (DF) Chest CT showed diffuse infiltration in both lungs, but the lesions significantly resolved after tocilizumab treatment. Figures A and D show a 55-year-old male patient whose primary symptoms were fever, cough, and sputum, and whose lesions resolved on day 5 after tocilizumab treatment. Figures B and E show an 82-year-old male patient whose primary symptoms were fever, and whose lesions resolved on day 5 after tocilizumab treatment. Figures C and F show a 32-year-old male patient whose primary symptoms were fever, cough, and sputum, and whose lesions resolved on day 4 after tocilizumab treatment.

[0052] Figure 6: CRP, body temperature, oxygen concentration, and oxygen saturation values ​​in 21 COVID-19 patients before and after tocilizumab treatment. (A) After tocilizumab treatment, C-reactive protein (CRP) significantly decreased and returned to normal in most patients (16 / 19, 84.2%). (B) After tocilizumab treatment, fever returned to normal in all 21 patients. (CD) Before treatment, 20 patients required oxygen therapy, including 9 patients (45.0%) receiving high-flow oxygen, 7 patients (35.0%) receiving nasal cannula, 1 patient (5.0%) receiving mask oxygen, 1 patient (5.0%) receiving non-invasive ventilation, and 2 patients (10.0%) receiving invasive ventilation. After tocilizumab treatment, 15 patients (75.0%) reduced their oxygen intake, and oxygen saturation remained stable. One patient did not require further oxygen therapy on the third day, so the oxygen concentration was marked as 21%, similar to the oxygen content in normal air. Figure 6B -D uses paired t-test analysis. Detailed Implementation

[0053] To make the objectives, technical solutions, and advantages of the present invention clearer, the present invention will be further described in detail below with reference to specific embodiments and accompanying drawings.

[0054] Unless otherwise specified, the methods used in the following examples are conventional methods, and the reagents used are commercially available reagents.

[0055] In a specific embodiment of the present invention, the treatment regimen for subjects with novel coronavirus is "tocilizumab + conventional treatment". The conventional treatment is defined according to the National Health Commission's "Notice on Issuing the Diagnosis and Treatment Protocol for Novel Coronavirus (Trial Version 5, Revised Edition)" (National Health Commission Medical Letter

[2020] No. 117). Specifically, the conventional treatment includes general treatment, treatment for severe and critically ill cases, and traditional Chinese medicine treatment.

[0056] In a specific embodiment of the present invention, "general treatment" includes the following:

[0057] 1. Bed rest, enhanced supportive treatment, and adequate calorie intake; pay attention to water and electrolyte balance and maintain internal environment stability; closely monitor vital signs, oxygen saturation, etc.

[0058] 2. Monitor blood routine, urine routine, CRP, biochemical indicators (liver enzymes, myocardial enzymes, kidney function, etc.), coagulation function, arterial blood gas analysis, and chest imaging according to the patient's condition. Cytokine testing may be performed if available.

[0059] 3. Provide timely and effective oxygen therapy, including nasal cannula, face mask oxygenation, and high-flow nasal cannula oxygen therapy.

[0060] 4. Antiviral therapy: Currently, there is no confirmed effective antiviral treatment. Alpha-interferon nebulized inhalation (5 million U or equivalent dose for adults, added to 2 ml of sterile water for injection, twice daily), lopinavir / ritonavir (200 mg / 50 mg per tablet), 2 tablets twice daily, or ribavirin (500 mg / dose, 2-3 times daily via intravenous infusion) can be added. Attention should be paid to adverse reactions associated with lopinavir / ritonavir, such as diarrhea, nausea, vomiting, and liver damage, as well as interactions with other drugs.

[0061] 5. Antibiotic treatment: Avoid blind or inappropriate use of antibiotics, especially the combined use of broad-spectrum antibiotics.

[0062] In specific embodiments of the present invention, the "treatment of severe and critically ill cases" generally includes the following methods:

[0063] 1. Treatment principles: On the basis of symptomatic treatment, actively prevent and treat complications, treat underlying diseases, prevent secondary infections, and provide timely organ function support.

[0064] 2. Respiratory support:

[0065] (1) Oxygen therapy: Severe patients should receive oxygen via nasal cannula or face mask, and timely assessment should be made to determine whether respiratory distress and / or hypoxemia have been relieved.

[0066] (2) High-flow nasal cannula oxygen therapy or non-invasive mechanical ventilation: When respiratory distress and / or hypoxemia cannot be relieved after standard oxygen therapy, high-flow nasal cannula oxygen therapy or non-invasive ventilation may be considered. If the condition does not improve or even worsens within a short period of time (1-2 hours), endotracheal intubation and invasive mechanical ventilation should be performed in a timely manner.

[0067] (3) Invasive mechanical ventilation: A lung-protective ventilation strategy is adopted, namely, mechanical ventilation with small tidal volume (4-8 ml / kg ideal body weight) and low inspiratory pressure (plateau pressure <30 cmH2O) to reduce ventilator-associated lung injury. Many patients experience patient-ventilator asynchrony, and sedation and muscle relaxants should be used in a timely manner.

[0068] (4) Salvage therapy: Lung recruitment is recommended for patients with severe acute respiratory distress syndrome (ARDS). Prone ventilation should be performed for at least 12 hours per day, provided there is sufficient manpower. If prone ventilation is ineffective, extracorporeal membrane oxygenation (ECMO) should be considered as soon as possible, if conditions permit.

[0069] 3. Circulatory support: On the basis of adequate fluid resuscitation, improve microcirculation, use vasoactive drugs, and perform hemodynamic monitoring if necessary.

[0070] 4. Other treatment measures

[0071] Depending on the severity of the patient's dyspnea and the progression of chest imaging, glucocorticoids may be used for a short period (3-5 days), with the recommended dose not exceeding the equivalent of methylprednisolone 1-2 mg / kg / day. It should be noted that higher doses of glucocorticoids may delay the clearance of the coronavirus due to their immunosuppressive effects. 100 ml of Xuebijing (a traditional Chinese medicine) may be administered intravenously twice daily. Intestinal microecological regulators may be used to maintain intestinal microecological balance and prevent secondary bacterial infections. Convalescent plasma therapy may be used. For critically ill patients with high inflammatory responses, extracorporeal blood purification technology may be considered if available.

[0072] In specific embodiments of the present invention, "traditional Chinese medicine treatment" generally includes the following methods:

[0073] 1. Medical observation period

[0074] Clinical manifestation 1: Fatigue accompanied by gastrointestinal discomfort

[0075] Recommended traditional Chinese medicine: Huoxiang Zhengqi capsules (pills, water, oral liquid)

[0076] Clinical manifestation 2: Fatigue accompanied by fever

[0077] Recommended traditional Chinese medicines: Jinhua Qinggan Granules, Lianhua Qingwen Capsules (Granules), Shufeng Jiedu Capsules (Granules), Fangfeng Tongsheng Pills (Granules)

[0078] 2. Clinical treatment period

[0079] (1) Initial stage: Cold and dampness stagnate in the lungs

[0080] Clinical manifestations: chills and fever or no fever, dry cough, dry throat, fatigue, chest tightness, epigastric fullness, or nausea and vomiting, loose stools. The tongue is pale or pale red with a white and greasy coating, and the pulse is soft.

[0081] Recommended prescription: Atractylodes lancea 15g, Citrus reticulata peel 10g, Magnolia officinalis 10g, Pogostemon cablin 10g, Amomum tsao-ko 6g, Ephedra sinica 6g, Notopterygium incisum 10g, Zingiber officinale 10g, Areca catechu 10g

[0082] (2) Mid-term: Epidemic virus closes the lungs

[0083] Clinical manifestations: persistent fever or alternating chills and fever, cough with little sputum or yellow sputum, abdominal distension and constipation. Chest tightness and shortness of breath, cough with wheezing, shortness of breath upon exertion. Red tongue with yellow and greasy or yellow and dry coating, and slippery and rapid pulse.

[0084] Recommended prescription: Apricot kernel 10g, gypsum 30g, Trichosanthes kirilowii 30g, raw rhubarb 6g (added later), raw and processed ephedra 6g each, Lepidium apetalum 10g, peach kernel 10g, Amomum tsao-ko 6g, areca nut 10g, Atractylodes lancea 10g

[0085] Recommended traditional Chinese medicines: Xiyanping Injection, Xuebijing Injection

[0086] (3) Severe stage: internal closure and external collapse

[0087] Clinical manifestations: difficulty breathing, shortness of breath with exertion or need for assisted ventilation, accompanied by coma, irritability, sweating and cold limbs, dark purple tongue, thick and greasy or dry tongue coating, and floating, large and weak pulse.

[0088] Recommended prescription: Ginseng 15g, Aconite root slices 10g (decocted first), Cornus officinalis 15g, taken with Suhexiang pills or Angong Niuhuang pills.

[0089] Recommended traditional Chinese medicines: Xuebijing Injection, Shenfu Injection, Shengmai Injection

[0090] (4) Recovery period: Lung and spleen qi deficiency

[0091] Clinical manifestations: shortness of breath, fatigue, poor appetite, nausea and vomiting, abdominal distension, weak bowel movements, loose stools, pale and swollen tongue with a white and greasy coating.

[0092] Recommended prescription: Pinellia ternata 9g, Citrus reticulata peel 10g, Codonopsis pilosula 15g, Astragalus membranaceus (processed) 30g, Poria cocos 15g, Pogostemon cablin 10g, Amomum villosum 6g.

[0093] Example 1 Subject CD4 + Expression of GM-CSF, IFN-γ and IL-6 in T cells

[0094] The proportion, absolute number, and phenotypic molecular characteristics of peripheral blood immune cells were detected by flow cytometry in 33 patients with novel coronavirus (including 12 ICU patients with severe symptoms and 21 patients with mild symptoms) and 10 healthy individuals.

[0095] Key factors of cytokine storm, including interferon-γ (IFNγ), colony-stimulating factor (GM-CSF), and IL-6, were detected. Results showed that, compared to healthy controls, patients with novel coronavirus had significantly higher levels of fresh CD4+. + T cells still produce high proportions of GM-CSF, IFN-γ, and IL-6, with even higher expression levels in severely ill ICU patients (see [link]). Figure 1 ).

[0096] Example 2: Expression of IL-6 in the monocytes of the subject

[0097] The proportion, absolute number, and phenotypic molecules of peripheral blood immune cells in the above 33 patients with novel coronavirus (including 12 ICU severe patients and 21 ordinary patients) and 10 normal individuals were detected by flow cytometry.

[0098] Monocytes are an important component of peripheral immune cells. Using flow cytometry with internal standard cytokines, IL-6, a major factor in cytokine storm, was detected. Compared with healthy controls, CD4+ was found to be significantly increased in patients with novel coronavirus. + Monocytes produce a high proportion of IL-6, and importantly, in severely ill ICU patients, these inflammatory monocytes also secrete high levels of IL-6, exacerbating the cytokine storm (see...). Figure 2 The above demonstrates that IL-6 is a key target for immunotherapy.

[0099] Example 3: Changes in biochemical indicators of subjects after administration of tocilizumab

[0100] Tocilizumab injection, a drug used clinically for rheumatic diseases, can block the interleukin-6 pathway. According to the National Health Commission's document No. 117 (2020), "Notice on Issuing the Diagnosis and Treatment Protocol for Novel Coronavirus (Trial Version 5, Revised Edition)," six patients from the First Affiliated Hospital of the University of Science and Technology of China and the Second People's Hospital of Fuyang, Anhui Province, were clinically classified, including one mild case and five severe cases. A new treatment regimen of "tocilizumab + conventional treatment" was adopted; the conventional treatment was defined according to the National Health Commission's document No. 117 (2020).

[0101] In this embodiment, 6 patients had undergone 4-10 days of routine treatment, 7 patients still had typical lesions in both lungs, and 6 patients all presented with high fever (average fever duration 7.8 days, average body temperature reaching 38.3℃). Among the 6 patients, 2 had white blood cell counts below the normal reference range (reference range 3.5-9.5, unit ×10). 9In 6 cases, all lymphocyte counts were below the normal reference range (reference range: 1.1-3.2, unit: ×10⁻⁶ / L). 9 The serum interleukin-6 (IL-6) levels of all 6 patients were higher than the normal reference range (reference range is <7, unit pg / ml), the serum C-reactive protein levels of all 6 patients were higher than the normal reference range (reference range is <8, unit mg / L), and the oxygenation index of all 6 patients was lower than the normal reference range (reference range 400-500, unit mmhg), as shown in Table 1.

[0102] After treatment with intravenous tocilizumab 400mg, the body temperature of 6 febrile patients returned to normal (temperature below or equal to 37℃) after 24 hours, and remained normal for 84 hours. Statistical analysis (paired t-test) showed significant differences in temperature changes. Figure 3 As shown. CT images of Case 7 before and after medication are shown below. Figure 4 As can be seen from the image, the patient's lung inflammation has significantly improved.

[0103] In addition, the changes in blood parameters of the 6 subjects 24 hours after drug administration are shown in Table 1. Five patients showed an increase in white blood cell and / or lymphocyte counts, indicating a recovery in immune function and improved antiviral capacity; six patients showed a decrease in C-reactive protein, indicating a decrease in inflammatory response and a reduction in cytokine storm; simultaneously, five patients showed improvement in oxygenation index (see Table 1). IL6 is a cytokine that requires its receptor (IL6 receptor) to exert its effector function. Tocilizumab targets the IL6 receptor, blocking the binding of IL6 to its receptor, thereby blocking IL6 function. After tocilizumab treatment, IL6 cannot bind to its receptor, and thus cannot be consumed, accumulating in the body, resulting in an increase in blood IL6 levels after treatment; however, even with this increase, the presence of tocilizumab in the body prevents it from exerting its effector function.

[0104] Table 1

[0105]

[0106] Example 4: Tocilizumab treatment for COVID-19 patients

[0107] A total of 21 patients participated in tocilizumab treatment. Seven patients received treatment at the First Affiliated Hospital of the University of Science and Technology of China (Anhui Provincial Hospital), and 14 patients received treatment at the Second People's Hospital of Fuyang, Anhui Province. All registered patients met the criteria outlined in the "Diagnosis and Treatment Protocol for Novel Coronavirus Pneumonia (Trial Version 6)" issued by the National Health Commission of the People's Republic of China.th The criteria for severe or critical illness as defined in the Interim Edition of the Chinese NHCOTPSRO (China National Health and Family Planning Commission) are as follows: A severe diagnosis is defined as meeting any of the following conditions in addition to being diagnosed with COVID-19: (1) respiratory rate ≥ 30 breaths / minute; (2) SpO2 ≤ 93% when breathing indoor air; (3) PaO2 / FiO2 ≤ 300 mmHg. A critical illness is diagnosed if any of the following conditions are present: (1) respiratory failure requiring mechanical ventilation; (2) shock; (3) concurrent failure of other organ functions requiring ICU admission.

[0108] The Medical Research Ethics Committee of Anhui Provincial Hospital approved the study. All patients signed informed consent forms before using tocilizumab and agreed to the publication of this case series. We are committed to protecting patient privacy and complying with the Declaration of Helsinki.

[0109] (Approval No.: 2020-XG (H)-015)

[0110] IL-6 test

[0111] IL-6 levels were measured using electrochemiluminescence immunoassay (Roche Diagnostics GmbH) or FACS analysis. Intracellular staining for IL-6 was performed without any restimulation. Cells were then collected, washed, and blocked according to eBioscience's instructions. The normal range for IL-6 is less than 7 pg / ml.

[0112] Treatment and observation

[0113] All patients received standard care according to the "Diagnosis and Treatment Protocol for Novel Coronavirus Infections" (Trial Version 6), including lopinavir, methylprednisolone, other symptom relievers, and oxygen therapy, plus tocilizumab (Roche Pharma (Schweiz) Ltd, B2084B21), administered as a single intravenous infusion of 400 mg. Clinical characteristics, including body temperature and oxygen saturation, were recorded. Complete blood counts were repeated. All patients underwent spiral computed tomography (CT) scans at admission and one week after initiation of tocilizumab treatment using a 64-slice spiral Optima CT680 scanner (GE Healthcare, USA) with 5 mm slices, performing whole-lung, low-dose exposure.

[0114] Data collection

[0115] Treatment data were collected through charts and graphs, including gender, age, comorbidities, clinical symptoms, and peripheral oxygen saturation. The study focused on changes in body temperature, respiratory function, and CT findings before and after tocilizumab treatment.

[0116] Statistical analysis

[0117] All statistical data were analyzed using IBM SPSS version 16.0 (Chicago, IL, USA) and are expressed as mean ± standard deviation.

[0118] result

[0119] Demographic characteristics

[0120] The mean age of the subjects was 56.8 ± 16.5 years, ranging from 25 to 88 years (Table 2). Of the 21 patients, 18 were male (85.7%) and 3 were female (14.3%). Seventeen patients (81.0%) were assessed as severe and four patients (19.0%) as critical. Of these, five patients (23.8%) had a history of exposure, and six patients (28.6%) had been exposed to a patient diagnosed with COVID-19. Eighteen patients (85.7%) received a single dose of tocilizumab, and three patients (14.3%) received a second dose of the same tocilizumab due to fever within 12 hours.

[0121] Clinical presentation

[0122] All patients presented with fever as the first symptom, followed by dry cough (14 / 21, 66.7%), copious white sputum (9 / 21, 42.9%), fatigue (6 / 21, 28.6%), and dyspnea (6 / 21, 28.6%). Nausea was present in 4 patients (19.0%). Other symptoms, including rhinorrhea and chest pain, were rare (4.8%). The median interval from the onset of fever to the development of dyspnea was 6 days (range 2–14 days) across all patients. Twenty patients required oxygen therapy, including 9 patients (45.0%) requiring high-flow oxygen, 7 patients (35.0%) requiring nasal cannula, 1 patient (5.0%) requiring face mask oxygen, 1 patient (5.0%) requiring non-invasive ventilation, and 2 patients (10.0%) requiring invasive ventilation. (Table 2)

[0123] Laboratory tests

[0124] White blood cell counts showed abnormal values ​​in peripheral blood in 4 patients (4 / 20, 20.0%) (mean 6.30 ± 2.77 x 10⁻⁶). 9 / L). (Table 3). 85.0% of patients had a decreased lymphocyte percentage (17 / 20, mean 15.52 ± 8.89%). C-reactive protein (CRP) levels were increased in all 20 patients (mean 75.06 ± 66.80 mg / L). The median procalcitonin (PCT) value was 0.33 ± 0.78 ng / mL, with only 2 patients (10.0%) showing abnormal values. Furthermore, prior to tocilizumab treatment, IL-6 expression levels were analyzed in all patients, with a mean of 132.38 ± 278.54 pg / ml, indicating upregulation of IL-6 in these severe and critically ill COVID-19 patients.

[0125] Imaging features

[0126] All patients presented with abnormal chest CT findings. The main abnormalities on the initial chest CT were patchy, ground-glass opacities and focal consolidation, primarily distributed in the periphery, especially the subpleural region. Figure 5 (AC). Within the first 7 days of admission, the size, extent, and severity of ground-glass opacities increased in 21 patients. No pleural effusion, mediastinal nodule, or central pulmonary embolism was found.

[0127] Treatment results

[0128] Following administration of tocilizumab, all patients' body temperatures returned to normal significantly on the first day and then remained stable. Figure 6B In the following days, clinical symptoms significantly improved. Peripheral oxygen saturation significantly improved. Figure 6C -D). One patient did not require further oxygen therapy. Fifteen patients (15 / 20, 75.0%) had their oxygen intake reduced. One patient was weaned off the ventilator on day one after tocilizumab treatment. One critically ill patient was extubated on day five and regained consciousness. Another patient who had previously received invasive ventilation had their ventilator parameters reduced and is expected to be weaned off the ventilator soon.

[0129] As shown in Table 3, Table 5-8 and Figure 6A As shown, lymphocyte percentage and CRP levels changed significantly after tocilizumab treatment. On day 5 post-treatment, only 2 patients (2 / 19, 10.5%) had abnormal white blood cell counts, with a mean of 5.25 ± 2.11 x 10⁻⁶. 9 / L. In 10 patients (10 / 19, 52.60%), the lymphocyte percentage returned to normal (mean 22.62 ± 13.48%). On day 5 post-treatment, CRP significantly decreased and returned to normal in 84.20% of patients (16 / 19, mean 2.72 ± 3.60 mg / L). Post-treatment CT scans showed lesion absorption in 19 patients (90.5%), with slight improvement in the others (see CT scans before and after treatment). Figure 5 AC and Figure 5 (DF). Nineteen patients (90.5%) have been discharged, including two critically ill patients. The remaining patients remain under observation in the hospital, but their body temperature remains normal (Table 4), and all symptoms have significantly improved. The mean length of hospital stay after tocilizumab treatment was 13.5 ± 3.1 days (Table 2). There were no reports of subsequent pulmonary infections, disease exacerbations, or deaths. No adverse drug reactions were reported during tocilizumab treatment.

[0130] discuss

[0131] In this study, all 21 patients had a one-week history of routine treatment prior to tocilizumab treatment, but their condition worsened with persistent fever, hypoxemia, and deteriorating CT images. Following tocilizumab treatment, in addition to improved body temperature (see Table 4), most patients experienced some degree of improvement in respiratory function. Chest tightness was relieved. In 15 patients (75.0%), oxygen intake flow was reduced, and oxygen saturation remained stable. Within 5 days of tocilizumab treatment, two patients were weaned off mechanical ventilation, and another critically ill patient is expected to be weaned off mechanical ventilation shortly.

[0132] A decrease in lymphocyte percentage is considered an important indicator for diagnosis and severity assessment in COVID-19 patients. In our study, we found that within 5 days of tocilizumab administration, lymphocyte percentage decreased in 85.0% of patients (17 / 20), and returned to normal in 52.6% (10 / 19). Simultaneously, elevated CRP levels also normalized. In 19 patients (90.5%), ground-glass opacities in the lungs observed on CT scans were absorbed. Considering the sufficient time required for lung tissue damage to repair, a delayed response on CT scans can be expected. No adverse drug reactions or subsequent pulmonary infections were reported during treatment. All patients experienced significant improvement in clinical symptoms and had a good prognosis after treatment. All 19 patients (90.5%) have been discharged, including 2 critically ill patients. Therefore, tocilizumab is effective in treating severe COVID-19 patients, likely due to the blockade of the IL-6-related fever and inflammatory storm response.

[0133] Table 2: Demographic characteristics of patients

[0134]

[0135]

[0136]

[0137] CHD, Coronary Artery Disease; COPD, Chronic Obstructive Pulmonary Disease; CKD, Chronic Kidney Disease. Unless otherwise stated, data are n / N (%).

[0138] * Additions and subtractions are average ± SD.

[0139] ** Patients who have been diagnosed with COVID-19.

[0140] ***Hospitalization for several days after tocilizumab treatment.

[0141] Table 3: Laboratory tests before and after tocilizumab treatment

[0142]

[0143] Note: Data are presented as mean ± SD (number of abnormalities / total number, %). Due to special clinical circumstances, some data could not be collected in a timely manner.

[0144] Table 4: Body temperature (°C) of patients before and after tocilizumab treatment

[0145]

[0146] Table 5: IL-6 levels before and after tocilizumab treatment

[0147]

[0148] Note: Due to the special circumstances of the pandemic, some data could not be collected in a timely manner.

[0149] Table 6: C-reactive protein levels before and after tocilizumab treatment

[0150]

[0151] Note: Due to the special circumstances of the pandemic, some data could not be collected in a timely manner.

[0152] Table 7: Lymphocyte counts before and after tocilizumab treatment

[0153]

[0154] Note: Due to the special circumstances of the pandemic, some data could not be collected in a timely manner.

[0155] Table 8: White blood cell counts before and after tocilizumab treatment

[0156]

[0157] The above results indicate that the IL-6 receptor antibody tocilizumab, especially 400 mg tocilizumab, achieves one or more of the following effects: a significant decrease in C-reactive protein; a significant reduction in fever response and a return to normal body temperature; an increase in the number of immune cells, specifically a significant increase in the number of peripheral leukocytes and lymphocytes, which is beneficial to the antiviral response; and a reduction in lung damage, specifically an improvement in the oxygenation index, demonstrating a good effect on the treatment of novel coronavirus.

[0158] The specific embodiments described above further illustrate the purpose, technical solution, and beneficial effects of the present invention. It should be understood that the above descriptions are merely specific embodiments of the present invention and are not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. Use of an IL-6 receptor antibody in a single-dose unit of 350 mg-450 mg in the preparation of a medicament or kit for treating patients with novel coronavirus, wherein the IL-6 receptor antibody is tocilizumab, and wherein the medicament or kit further comprises an antiviral agent, wherein when the IL-6 receptor antibody is present in the kit, the IL-6 receptor antibody is packaged separately from the antiviral agent. The antiviral agents mentioned are alpha-interferon, lopinavir / ritonavir, or ribavirin.

2. The use as described in claim 1, wherein the patient is a mild patient, a moderate patient, a severe patient, or a critically ill patient.

3. The use according to any one of claims 1-2, wherein the drug or kit further comprises a traditional Chinese medicine preparation, wherein the traditional Chinese medicine preparation is selected from Xiyanping injection, Xuebijing injection, Shenfu injection and / or Shengmai injection.

4. The use according to claim 3, wherein the IL-6 receptor antibody is packaged separately from the antiviral preparation and the traditional Chinese medicine preparation.

5. The use according to claim 1, wherein the antiviral agent is α-interferon with a single dose unit of 5 million U, lopinavir with a single dose unit of 200 mg, ritonavir with a single dose unit of 50 mg, or ribavirin with a single dose unit of 500 mg.

6. The use according to claim 5, wherein the single dose of α-interferon is 5 million U twice daily; the single dose of lopinavir / ritonavir is 200 mg / 50 mg per tablet twice daily; and the single dose of ribavirin is 500 mg / dose, 2-3 times daily.

7. The use according to claim 1, wherein, The single dose of tocilizumab is 350 mg, 360 mg, 370 mg, 380 mg, 390 mg, 400 mg, 410 mg, 420 mg, 430 mg, 440 mg or 450 mg.

8. The use as claimed in claim 1, wherein the tocilizumab is in a form suitable for intravenous infusion.

9. The use according to claim 1, wherein the drug or kit further comprises a glucocorticoid.

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