Application of cobalt complex in preparation of medicine for treating cytokine storm related diseases

Abstract

The invention discloses application of a cobalt complex in preparation of a medicine for treating cytokine storm related diseases. A large number of natural medicines with anti-inflammatory effects are verified, then based on structural formulas of the natural medicines, a machine learning algorithm is combined to assist medicine design, some small molecule medicines are artificially synthesized, and the anti-inflammatory effects of the medicines are verified at a cellular level and an animal level and are subjected to target exploration; finally, it is found that the cobalt complex Co68 has a remarkable anti-inflammatory effect and has a wide application prospect in the field of treatment of cytokine storm related diseases.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology, and in particular relates to the application of cobalt complexes in the preparation of drugs for treating cytokine storm-related diseases. Background Art

[0002] Cytokine storm, also known as cytokine release syndrome (CRS), is a serious immune response disease. It is characterized by the abnormal release of a large number of inflammatory cytokines by the body, triggering systemic inflammatory responses and tissue damage. Usually, cytokine storms are caused by the loss of control of the immune system caused by infection, immunotherapy or other stimuli. Its clinical manifestations are varied, including symptoms such as high fever, chills, increased heart rate, shortness of breath, hypotension, and tissue edema. In severe cases, serious complications such as shock and multiple organ failure may occur, seriously threatening the patient's life. Although the pathogenesis of cytokine storms has not yet been fully clarified, studies have shown that it is closely related to the excessive release of multiple inflammatory cytokines, such as tumor necrosis factor (TNF), interleukin-6 (IL-6), and interleukin-1β (IL-1β). These inflammatory cytokines can induce systemic inflammatory responses, leading to serious consequences such as increased vascular permeability, thrombosis, and organ dysfunction. For example, the SARS-CoV-2 virus causes a cytokine storm reaction in the patient's body, which poses a serious threat to the patient's life and health. Therefore, timely diagnosis and effective treatment of cytokine storm-related diseases are crucial to improving the survival rate of patients.

[0003] Methods for treating cytokine storms mainly include controlling inflammatory responses, maintaining hemodynamic stability, and supporting organ function. Common treatment measures include anti-inflammatory drugs, vasoactive drugs, fluid resuscitation, and respiratory support. However, currently available treatments such as immunomodulators or cytokine inhibitors are quite limited and are limited to treating cytokine storms caused by specific infections, and are not broad-spectrum.

[0004] Metal elements are important components of life, and metal ions and metal proteins play an important role in many life processes. For example, among all known enzymes, about one-third of them have metal cofactors, which constitute the structure of the enzyme, catalytic components or regulate enzyme functions. In fact, people have long been studying the regulation of the immune system by metal ions, such as the discovery of various metal allergies. The large-scale emergence of metal nickel contact dermatitis in 1950 is the most common and famous hypersensitivity reaction in the world. Nickel ions activate macrophages by binding to TLR4, releasing inflammatory factors and type I interferon and stimulating T cell immunity. Another example is aluminum adjuvant, which was discovered in 1926 and began to be used worldwide. In recent years, there have been more and more reports on the functions of metal elements in the immune system, which has deepened people's understanding of metal immunology. For example, according to research, manganese ions can directly activate the innate immune signaling pathway cGAS-STING-TBK1-IRF3-IFNB1 by binding to cGAS, which has clinical application value in antiviral and anti-tumor. As research on metal immunology continues to deepen, it is expected to reveal more complex and subtle interactions between metal elements and the immune system, providing new insights and solutions for human health and disease prevention and treatment.

[0005] Although it has been reported that several common metal elements such as manganese, calcium, iron, cobalt, nickel and zinc directly participate in or indirectly regulate the function of the immune system, the specific mechanism of action of metal complexes in mediating immune and inflammatory responses is still unclear. Therefore, it is still necessary to study the specific mechanism of action of metal complexes in mediating immune and inflammatory responses in order to obtain new strategies for treating cytokine storm-related diseases. Summary of the invention

[0006] In order to solve at least some of the technical problems in the above-mentioned prior art, the present invention verifies a large number of natural medicines with anti-inflammatory effects, and then based on the structural formulas of these natural medicines, combines machine learning algorithms to assist drug design, and finally finds that the cobalt complex Co68 has excellent effects in treating cytokine storm-related diseases. Specifically, the present invention includes the following contents.

[0007] In a first aspect of the present invention, a cobalt complex Co68 is provided for use in the preparation of a drug for treating cytokine storm-related diseases, wherein the structural formula of the cobalt complex Co68 is as shown in Formula I:

[0008]

[0009] In certain embodiments, the use of the cobalt complex Co68 according to the present invention in the preparation of a drug for treating a cytokine storm-related disease, wherein the tBu The PNP ligand and the cobalt salt react to obtain the cobalt complex Co68.

[0010] In certain embodiments, the cobalt complex Co68 according to the present invention is used in the preparation of a medicament for treating cytokine storm-related diseases, wherein the cytokines include at least one of TNF, IL-1, IL-2, IL-4, IL-6, CSF2, CSF3, IL-27, LTb, IL-8, IL-10, IL-12, IL-17, IL-18, IFN-γ, IFN-α, IP-10, CXCL10, CCL7, GM-CSF, MCP-1, MIP-1α and MIP-1β.

[0011] In certain embodiments, the cobalt complex Co68 according to the present invention is used in the preparation of a drug for treating cytokine storm-related diseases, wherein the cytokine storm-related diseases include at least one of infectious diseases, autoimmune diseases, AIDS and neurodegenerative diseases.

[0012] In certain embodiments, the use of the cobalt complex Co68 according to the present invention in the preparation of a medicament for treating a cytokine storm-related disease, wherein the treatment includes at least one of the following situations:

[0013] (1) Reduce the amount of cytokines;

[0014] (2) Inhibit inflammatory response;

[0015] (3) inhibiting inflammatory response signaling pathways;

[0016] (4) Activation of innate immune cytokines;

[0017] (5) Prolong survival time.

[0018] In certain embodiments, according to the use of the cobalt complex Co68 described in the present invention in the preparation of a drug for treating cytokine storm-related diseases, the inflammatory response signaling pathway includes at least one of a TNF signaling pathway, a TLR signaling pathway, and a JAK-STAT signaling pathway.

[0019] In certain embodiments, according to the use of the cobalt complex Co68 described in the present invention in the preparation of a medicament for treating cytokine storm-related diseases, wherein the natural immune cytokines include at least one of IFNA2, IFNA4, IFNB1, EGR1, TRIM172 and DUSP27.

[0020] The second aspect of the present invention provides a method for regulating cytokine levels in vitro, which comprises the step of contacting the cobalt complex Co68 according to the present invention with cells, wherein the regulation comprises reducing the amount of cytokines or inhibiting the production of cytokines.

[0021] In certain embodiments, according to the method for regulating cytokine levels in vitro according to the present invention, the cytokines include at least one of TNF, IL-1, IL-2, IL-4, IL-6, CSF2, CSF3, IL-27, LTb, IL-8, IL-10, IL-12, IL-17, IL-18, IFN-γ, IFN-α, IP-10, CXCL10, CCL7, GM-CSF, MCP-1, MIP-1α and MIP-1β.

[0022] The third aspect of the present invention provides the use of the cobalt complex Co68 in the preparation of a drug for treating cytokine storm-related diseases in combination with other drugs, wherein the structural formula of the cobalt complex Co68 is as shown in Formula I:

[0023]

[0024] The present invention verifies a large number of natural medicines with anti-inflammatory effects, and then based on the structural formulas of these natural medicines, combines machine learning algorithms to assist in drug design, and artificially synthesizes some small molecule drugs to verify their anti-inflammatory effects and explore targets at the cellular and animal levels. It is finally found that the cobalt complex Co68 has significant anti-inflammatory effects and has broad application prospects in the treatment of cytokine storm-related diseases.

[0025] The present invention has found through research that the cobalt complex Co68 has the following beneficial effects:

[0026] (1) Co68 has a broad spectrum of inhibitory effects on inflammatory responses at the cellular level, including inflammatory responses induced by the bacterial product LPS, as well as inflammatory responses caused by viral infections such as VSV and EMCV;

[0027] (2) Co68 can alleviate and slow down LPS-induced inflammation-related diseases at the animal level and significantly prolong the lifespan of critically ill mice in the LPS-induced sepsis model;

[0028] (3) Co68 can significantly inhibit typical cytokines in cytokine storm at both the transcriptional and protein levels, including IL1a, IL1b, IL-6, CSF2, CSF3 and other cytokines; and can significantly activate antiviral-related cytokines and their downstream genes and proteins in natural immunity, including IFNA2, IFNA4, IFNB1, EGR1, TRIM17 and DUSP27; and induce the expression of epigenetic-related genes, including H1F5, H2AC7, H2BC4 and H2BC18;

[0029] (4) The gene changes induced by Co68 were mainly enriched in the signaling pathways related to inflammatory response and innate immune response, including biological processes such as systemic lupus erythematosus, TNF signaling pathway, TLR signaling pathway, JAK-STAT signaling pathway, and rheumatoid arthritis;

[0030] (5) Co68 can significantly activate the innate immune response in different cell lines in a time-dependent manner;

[0031] (6) Co68 activates IFN by activating the TLR4 receptor, and it is dependent on the TRIF-TBK1-IRF3 classical signaling pathway downstream of TLR4 to activate the body's innate immune response;

[0032] (7) Co68 activates STAT1 through the adaptor protein downstream of TLR4, thereby exerting a broad-spectrum anti-inflammatory function;

[0033] (8) Co68 activates the TLR4-TBK1-IRF3-IFN signaling pathway while reshaping the body's immune and inflammatory responses by inhibiting TAK1-IKBa-P65 signaling. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] Figure 1 The results showed that Co68 has a broad spectrum of anti-inflammatory effects at the cellular and animal levels.

[0035] Figure 2 It was shown that Co68 can broadly inhibit lipopolysaccharide (LPS)-induced inflammatory cytokines.

[0036] Figure 3 This shows that Co68 can activate the body's natural immune response.

[0037] Figure 4 This showed that Co68 has antiviral function at the cellular level.

[0038] Figure 5 It was shown that Co68 activated the innate immune response through TLR4 and inhibited the inflammatory response through STAT1. DETAILED DESCRIPTION

[0039] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0040] It should be understood that the terms described in the present invention are only for describing special embodiments and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that the upper and lower limits of the scope and each intermediate value therebetween are specifically disclosed. Each smaller range between the intermediate value in any stated value or stated range and any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0041] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as commonly understood by those of ordinary skill in the art to which the invention belongs. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials related to the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0042] application

[0043] In one aspect of the present invention, a cobalt complex Co68 is provided for use in the preparation of a drug for treating cytokine storm-related diseases, wherein the structural formula of the cobalt complex Co68 is as shown in Formula I:

[0044]

[0045] The cobalt complex Co68 used in the present invention can be prepared by the method described in Chinese patent application CN107964028A, the content of which is incorporated herein by reference in its entirety. Specifically, the cobalt complex in the present invention is prepared by the following method:

[0046] will dissolve tBu A solution of PNP ligand (0.361 g, 1.0 mmol) in tetrahydrofuran (THF) (5 ml) was added to a solution of cobalt dichloride (0.115 g, 0.90 mmol) in THF (5 ml). After the addition of the ligand, the reaction was stirred at room temperature (25°C) for 10 hours, and then the solution was concentrated to 2 ml, 15 ml of degassed n-hexane was added, and after stirring for 1 hour, argon was filtered, and the filter cake was dried to obtain a blue solid. The cobalt complex Co68 can be analyzed and verified by nuclear magnetic resonance and infrared determination. The cobalt complex Co68 used in the present invention tBu The PNP ligand is bis[2-(di-tert-butylphosphino)ethyl]amine, and the reaction equation is as follows:

[0047]

[0048] In the present invention, the cytokines (also referred to as "inflammatory cytokines") include but are not limited to at least one of TNF, IL-1, IL-2, IL-4, IL-6, CSF2, CSF3, IL-27, LTb, IL-8, IL-10, IL-12, IL-17, IL-18, IFN-γ, IFN-α, IP-10, CXCL10, CCL7, GM-CSF, MCP-1, MIP-1α and MIP-1β.

[0049] In the present invention, the cytokine storm-related diseases include but are not limited to infectious diseases, autoimmune diseases, AIDS, neurodegenerative diseases, etc.

[0050] In the present invention, the infectious diseases include but are not limited to diseases caused by bacterial infection and diseases caused by viral infection. Examples of the infectious diseases include but are not limited to sepsis, pneumonia, malaria, avian influenza, smallpox, pandemic influenza, adult respiratory distress syndrome (ARDS), severe acute respiratory syndrome (SARS), etc. The bacteria include but are not limited to Streptococcus pneumoniae, Pneumocystis jiroveci, Bordetella pertussis, Blastomyces dermatitidis, Cryptococcus, Coccidioides, Histoplasma capsulatum, Cryptococcus neoformans, Streptococcus pyogenes, Staphylococcus aureus, Klebsiella pneumoniae, etc. The viruses include DNA viruses and / or RNA viruses, and examples of viruses include, but are not limited to, Rhabdoviridae viruses (such as, but not limited to, vesicular stomatitis virus, VSV), encephalomyocarditis virus (EMCV), herpes simplex virus (HSV), rhinovirus, coronavirus, paramyxoviridae, orthomyxoviridae, adenovirus, parainfluenza virus, metapneumovirus, respiratory syncytial virus, influenza virus, Arenaviridae, Filoviridae, Bunyaviridae, Flaviviridae, Ebola virus, Marburg virus, Crimean-Congo hemorrhagic fever (CCHF), South American hemorrhagic fever, dengue fever, yellow fever, Rift Valley fever, Omsk hemorrhagic fever virus, Kwasana Forest disease, Junin virus, Machupo virus, Sabia virus, Guanarito virus, Garissa virus, Ilesa virus, Lassa fever virus, etc.

[0051] In the present invention, the autoimmune diseases include but are not limited to autoimmune neutropenia, Guillain-Barré syndrome, epilepsy, autoimmune encephalitis, Isaacs syndrome, nevus syndrome, pemphigus vulgaris, pemphigus decidua, bullous pemphigoid, acquired bullous epidermolysis bullosa, pemphigus gestationis, pemphigoid, mucous membrane pemphigoid, antiphospholipid syndrome, autoimmune anemia, autoimmune Graves' disease, thyroid eye disease (TED), Goodpasture's syndrome, myasthenia gravis, multiple sclerosis, rheumatoid arthritis, systemic lupus erythematosus, idiopathic thrombocytopenic purpura (ITP), warm antibody autoimmune hemolytic anemia (WAIHA), chronic inflammatory demyelinating polyneuropathy (CIDP), lupus nephritis, IgA nephropathy (Berger's disease), dermatomyositis, necrotizing autoimmune myopathy, membranous nephropathy, etc.

[0052] In the present invention, the neurodegenerative diseases include but are not limited to Alzheimer's disease, frontotemporal dementia (FTD), dementia, dementia with Lewy bodies (DLB), post-traumatic stress disorder, Parkinson's disease, vascular dementia, vascular cognitive impairment, Huntington's disease, multiple sclerosis, Creutzfeldt-Jakob disease, multiple system atrophy, progressive supranuclear palsy, etc.

[0053] In the present invention, the treatment includes at least one of the following situations:

[0054] (1) Reduce the amount of cytokines, including inhibiting the production and release of cytokines;

[0055] (2) Inhibit inflammatory response;

[0056] (3) inhibiting inflammatory response signaling pathways;

[0057] (4) Activation of innate immune cytokines;

[0058] (5) Prolong survival time.

[0059] In the present invention, the term "treatment" refers to therapeutic treatment or preventive measures, the purpose of which is to prevent or slow down (reduce) undesirable physiological changes or disorders, such as the occurrence and progression of inflammation. Beneficial or desired clinical results include, but are not limited to, the following results, whether detectable or undetectable, including relief of symptoms, reduction in disease severity, stabilization of the disease state (i.e., no worsening), delay or slowing of disease progression, improvement or alleviation of the disease state, and relief (whether partial or complete). Those in need of treatment include those who already have a cytokine storm-related disease or those who need to prevent or improve a cytokine storm-related disease.

[0060] In a preferred embodiment, the inflammatory response signaling pathway includes at least one of a TNF signaling pathway, a TLR signaling pathway, and a JAK-STAT signaling pathway.

[0061] In a preferred embodiment, the innate immune cytokine includes at least one of IFNA2, IFNA4, IFNB1, EGR1, TRIM172 and DUSP27.

[0062] method

[0063] In one aspect of the present invention, a method for regulating cytokine levels in vitro is provided, comprising the step of contacting the cobalt complex Co68 according to the present invention with cells, wherein the regulation comprises reducing the amount of cytokines or inhibiting the production of cytokines. The in vitro method of the present invention can be used for the study of the pathogenesis of cytokine storm-related diseases, drug screening and optimization, etc.

[0064] Joint Application

[0065] One aspect of the present invention provides the use of the cobalt complex Co68 according to the present invention in the preparation of a drug for treating cytokine storm-related diseases in combination with other drugs.

[0066] In the present invention, the other drugs include but are not limited to anti-inflammatory drugs, antiviral drugs, microbial inhibitors, vasoactive drugs, immunomodulators, cytokine inhibitors, etc.

[0067] Example

[0068] This example shows the role of the cobalt complex Co68 in inhibiting inflammatory responses and activating immune responses.

[0069] 1. Co68 has a broad spectrum of anti-inflammatory effects at the cellular and animal levels

[0070] 1.1 Cellular level

[0071] 1.1.1 Experimental methods

[0072] (1) The expression levels of target genes IL1a and IL1b were detected after the RAW 264.7 macrophage cell line was treated for 3 hours under different treatment conditions. The specific treatment conditions included: DMSO, LPS, LPS pretreatment for 15 minutes before applying Co68, LPS and Co68 treatment at the same time, Co68 pretreatment for 15 minutes before applying LPS, and Co68 alone.

[0073] (2) RAW 264.7 macrophage cell line was treated with VSV or HSV+Co68 for 24 hours, and the expression of inflammatory factor IL1b was detected.

[0074] (3) RAW 264.7 macrophage cell line was treated with EMCV, HSV, EMCV+Co68, and HSV+Co68 for 24 hours, and the expression of inflammatory factor IL1b was detected.

[0075] 1.1.2 Experimental Results

[0076] The results showed that Co68 could significantly inhibit the expression of IL1a induced by LPS ( Figure 1 Similarly, Co68 could significantly inhibit the expression of IL1b induced by LPS ( Figure 1 B).

[0077] After 24 hours of treatment with VSV alone, the expression of the inflammatory factor IL1b in the RAW 264.7 macrophage cell line was significantly increased. However, after 24 hours of co-treatment with VSV and Co68, VSV-induced viral inflammation was significantly inhibited ( Figure 1 C).

[0078] After 24 hours of treatment with EMCV and HSV alone, the expression of the inflammatory factor IL1b in the RAW 264.7 macrophage cell line was significantly increased. However, after 24 hours of simultaneous treatment with Co68, the viral inflammation induced by EMCV and HSV was also significantly inhibited ( Figure 1 D).

[0079] 1.2 Animal level

[0080] 1.2.1 Experimental methods

[0081] After the mice were injected with LPS at a dose of 15 mg / kg, PBS and Co68 solution were administered by intraperitoneal injection, respectively.

[0082] Mice were injected with LPS at a dose of 30 mg / kg, and PBS and Co68 solution were given by intraperitoneal injection, respectively.

[0083] 1.2.2 Experimental Results

[0084] The results showed that among mice treated with 15 mg / kg LPS, the survival time of mice treated with Co68 was significantly prolonged compared with mice treated with PBS alone ( Figure 1 In mice treated with 30 mg / kg LPS, the survival time of mice treated with Co68 was significantly prolonged compared with mice treated with PBS alone, and the difference was more obvious ( Figure 1 F).

[0085] 2. Co68 broadly inhibits LPS-induced inflammatory cytokines

[0086] 2.1 Experimental methods

[0087] RAW 264.7 macrophage cell line was treated with Co68, Co68+LPS and LPS for 3 hours, respectively, and the expressions of inflammatory cytokines IL1a, IL1b, IL-6, IL-27 and CSF3 and the expression of immune regulatory transcription factor Egr1 were detected.

[0088] 2.2 Experimental Results

[0089] The results showed that Co68 could not only significantly reduce the expression of IL1a and IL1b ( Figure 2 A and B), and could also significantly inhibit the expression of IL-6, IL-27 and CSF3 ( Figure 2 CE). At the same time, it was unexpectedly found that Co68 could significantly induce the expression of the innate immune regulatory transcription factor EGR1 ( Figure 2 F).

[0090] 3. Study on the activation of the body's natural immune response by Co68

[0091] 3.1 Experimental methods

[0092] In order to further verify that Co68 can activate innate immunity, RNA-seq analysis was performed in this example.

[0093] 3.2 Experimental Results

[0094] The results show that if Figure 3 As shown in A, compared with LPS treatment, Co68 significantly induced the expression of some genes related to innate immunity, including IFNA4, IFNB, EGR1, and TRIM17. At the same time, CO68 also significantly upregulated some genes related to epigenetics, such as H2BC18, H2BC4, H2AC7, and H1F5. In addition, consistent with the above inhibition experimental results, Co68 significantly downregulated the expression of several inflammatory cytokines, such as IL1b, IL1a, IL-27, CSF2, and LTb.

[0095] Figure 3 B shows a heat map of differentially expressed genes under Co68 and LPS treatment conditions, reflecting the changes in gene expression under different treatment conditions.

[0096] In protein interaction network analysis (PPI network), such as Figure 3As shown in C, most of the upregulated genes were enriched in the innate immune response pathway. Gene set enrichment analysis (GSEA) of the differentially expressed genes induced by Co68 revealed that these genes were mainly enriched in multiple immune-related signaling pathways, including the TNF signaling pathway, the Toll-like receptor signaling pathway, the JAK-STAT signaling pathway, the NOD-like receptor signaling pathway, and the cytoplasmic DNA recognition signaling pathway. This shows that Co68 can activate the innate immune response by regulating the above key signaling pathways.

[0097] 4.Co68 has antiviral function at the cellular level

[0098] 4.1 Experimental methods

[0099] In order to further verify the antiviral function of Co68, this example systematically evaluated its immunological properties.

[0100] 4.2 Experimental Results

[0101] like Figure 4 As shown in A, in the RAW 264.7 macrophage cell line, the activation effect of Co68 reached a peak after approximately 3 hours of treatment.

[0102] Next, this example evaluated the antiviral effect of Co68 in the RAW 264.7 macrophage cell line. Figure 4 As shown in Figure B, over time, the viral replication of VSV in the DMSO group gradually increased, while after Co68 treatment, the replication of the viral RNA fragment VSVg was significantly inhibited. To further verify this finding, similar experiments were conducted in the human colorectal adenocarcinoma cell line HT29 in this example, and the results further confirmed the significant inhibitory effect of Co68 on viral replication ( Figure 4 C).

[0103] Finally, the above results were confirmed at the protein level. Figure 4 As shown in D and E, after Co68 treatment, the phosphorylation levels of TBK1 and IRF3, key signaling proteins of innate immunity, increased significantly, further indicating its antiviral activity. At the same time, the important inflammatory factor transcription factor P65 was hardly phosphorylated after Co68 treatment, and its upstream inhibitory protein IKBα did not show degradation under Co68 treatment. Relatively speaking, under LPS treatment, the expression level of P65 increased over time, and IKBα also began to degrade after LPS treatment. These results collectively indicate that Co68 exerts its antiviral effect by regulating key immune pathways.

[0104] 5. Co68 activates innate immune response through TLR4 and inhibits inflammatory response through STAT1

[0105] 5.1 Experimental Methods

[0106] In order to explore the molecular mechanism by which Co68 activates innate immunity and simultaneously inhibits inflammatory response, this example used a variety of innate immune signaling pathway inhibitors, including STING inhibitor C176, TLR2 inhibitor C29, TLR4-MD2 co-inhibitor, TRIF inhibitor, MyD88 inhibitor and TBK1 inhibitor for target exploration.

[0107] 5.2 Experimental Results

[0108] The results show that if Figure 5 As shown in A, inhibitors of TLR4, TRIF, and TBK1 were able to significantly inhibit Co68-activated IFNB. Further validation in J774 and HT29 cell lines ( Figure 5 BC) also showed that the process of Co68 activating IFNB depends on the TLR4 signaling pathway.

[0109] Subsequently, bone marrow macrophages (BMDMs) from TLR4 knockout (KO) mice were used for experiments ( Figure 5 The results showed that Co68 could not activate IFNB in ​​TLR4 KO cells, which further confirmed that Co68 induced IFNB expression by activating TLR4 receptor. In addition, inhibitor and gene knockdown experiments ( Figure 5 The EF of the study verified the mechanism by which Co68 exerts a broad-spectrum anti-inflammatory function through the transcription factor STAT1. This shows that Co68 plays a key role in stimulating innate immunity and regulating inflammatory responses.

[0110] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit them; although the present invention has been described in detail with reference to the above embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the above embodiments, or replace some of the technical features therein by equivalents. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. The use of cobalt complex Co68 in the preparation of a drug for treating cytokine storm-related diseases, characterized in that: The structural formula of the cobalt complex Co68 is shown in Formula I:

2. The use of the cobalt complex Co68 according to claim 1 in the preparation of a drug for treating cytokine storm-related diseases, characterized in that: By making tBu The PNP ligand and the cobalt salt react to obtain the cobalt complex Co68.

3. The use of the cobalt complex Co68 according to claim 1 in the preparation of a drug for treating cytokine storm-related diseases, characterized in that: The cytokines include at least one of TNF, IL-1, IL-2, IL-4, IL-6, CSF2, CSF3, IL-27, LTb, IL-8, IL-10, IL-12, IL-17, IL-18, IFN-γ, IFN-α, IP-10, CXCL10, CCL7, GM-CSF, MCP-1, MIP-1α and MIP-1β.

4. The use of the cobalt complex Co68 according to claim 1 in the preparation of a drug for treating cytokine storm-related diseases, characterized in that: The cytokine storm-related diseases include at least one of infectious diseases, autoimmune diseases, AIDS and neurodegenerative diseases.

5. Use of the cobalt complex Co68 according to claim 1 in the preparation of a drug for treating cytokine storm-related diseases, characterized in that: The treatment includes at least one of the following: (1) Reduce the amount of cytokines; (2) Inhibit inflammatory response; (3) inhibiting inflammatory response signaling pathways; (4) Activation of innate immune cytokines; (5) Prolong survival time.

6. Use of the cobalt complex Co68 according to claim 5 in the preparation of a drug for treating cytokine storm-related diseases, characterized in that: The inflammatory response signaling pathway includes at least one of a TNF signaling pathway, a TLR signaling pathway and a JAK-STAT signaling pathway.

7. Use of the cobalt complex Co68 according to claim 5 in the preparation of a drug for treating cytokine storm-related diseases, characterized in that: The innate immune cytokine includes at least one of IFNA2, IFNA4, IFNB1, EGR1, TRIM172 and DUSP27.

8. A method for regulating cytokine levels in vitro, characterized in that: The method comprises the step of contacting a cobalt complex Co68 with cells, wherein the regulation comprises reducing the amount of cytokines or inhibiting the production of cytokines, and the structural formula of the cobalt complex Co68 is shown in Formula I:

9. The method for regulating cytokine levels in vitro according to claim 8, characterized in that: The cytokines include at least one of TNF, IL-1, IL-2, IL-4, IL-6, CSF2, CSF3, IL-27, LTb, IL-8, IL-10, IL-12, IL-17, IL-18, IFN-γ, IFN-α, IP-10, CXCL10, CCL7, GM-CSF, MCP-1, MIP-1α and MIP-1β.

10. Use of the cobalt complex Co68 in the preparation of a drug for treating cytokine storm-related diseases in combination with other drugs, characterized in that: The structural formula of the cobalt complex Co68 is shown in Formula I:

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