Stimulation of TRPV1 + sensory nerves for prevention and treatment of diseases

Abstract

The invention discloses prevention and treatment of diseases by stimulating TRPV1 + sensory nerves. Specifically, the invention discloses application of a substance for stimulating TRPV1 + sensory nerves in preparation of products for prevention, treatment or adjuvant treatment of inflammatory diseases, autonomic nerve dysfunction diseases or adrenal insufficiency diseases, and application of the substance for stimulating TRPV1 + sensory nerves in preparation of products for inhibiting proinflammatory cytokines and products for promoting anti-inflammatory cytokines. The TRPV1 ion channel agonist disclosed by the invention can obviously and effectively inhibit the expression of numerous proinflammatory cytokines and obviously improve the expression of anti-inflammatory cytokines IL10, so that not only is the physiological status of an inflammatory mouse obviously improved, but also the survival rate of the inflammatory mouse is obviously improved; meanwhile, the spleen function can be adjusted, expression of spleen pro-inflammatory genes is remarkably inhibited, generation of catecholamine is increased, and secretion of cortisol hormones is promoted. The invention has wide clinical application value in prevention and treatment of related diseases.

Description

Technical Field

[0001] This invention belongs to the field of biomedicine and relates to the prevention and treatment of diseases by stimulating TRPV1+ sensory nerves. Specifically, it relates to methods and drugs for stimulating TRPV1+ sensory nerves to regulate the autonomic nervous system and organ function to treat visceral dysfunction, regulate the secretion of hormones and catecholamines, and prevent and treat inflammatory diseases caused by pro-inflammatory cytokines or pro-inflammatory cytokine cascade reactions. Background Technology

[0002] Autonomic nervous system (vegetative nervous system) dysfunction, adrenal insufficiency, and imbalances in hormone and catecholamine secretion can disrupt the body's physiological homeostasis, leading to discomfort and inducing various diseases. Pathogen invasion, tissue damage, visceral dysfunction, autoimmune diseases, organ transplantation, immunotherapy, and exposure to stimulants can all trigger inflammatory responses. The immune system helps the body resist pathogen invasion, the proliferation of abnormal cells, and repair damaged tissues. However, immune system dysfunction leading to excessive and persistent inflammation will produce excessive pro-inflammatory cytokines and form a cytokine cascade. If the inflammatory process is not stopped in time, it will eventually lead to a cytokine storm or inflammatory storm. When this exceeds the body's normal compensatory anti-inflammatory response capacity, the immune system will indiscriminately attack the body's own tissues and cells, causing widespread damage to various systems and tissues throughout the body. In severe cases, it can lead to multiple organ failure and even death. Sepsis or septicemia caused by pathogenic microorganisms or other pathogenic factors has a high incidence rate, with over ten million severe cases worldwide each year. These two diseases are often accompanied by cytokine storms, leading to extremely severe illness and a mortality rate as high as 30%–70%, exceeding that of myocardial infarction. Critical illness or death caused by the coronaviruses SARS-CoV, MERS-CoV, and SARS-CoV-2, which have ravaged the world, is also closely related to cytokine storms. Cytokine storms release pro-inflammatory substances into the lung microenvironment in a short period, causing severe lung tissue damage. The concentration of pro-inflammatory cytokines in the plasma of severely ill patients is significantly higher than that of mildly ill patients, indicating a close correlation between symptom severity and high levels of cytokine expression. Immunotherapy and CAR-T cell therapy for cancer patients also carry a significant risk of inducing cytokine storms, even endangering their lives. For critically ill patients with the aforementioned diseases, while timely elimination of the source of the disease through antibiotics, antiviral drugs, or surgery is necessary, a key focus should be on effectively suppressing excessive inflammation. Furthermore, the persistent damage caused by chronic inflammation and autoimmune diseases should not be underestimated, often leading to various chronic diseases (such as diabetes and atherosclerosis) and cancer. Unfortunately, the available drugs and treatments for suppressing inflammation are currently limited, their efficacy is unsatisfactory, and they frequently cause serious side effects.

[0003] Besides the immune system, the nervous system also plays a crucial role in regulating inflammatory responses. Communication between neurons and immune cells promotes the functional integration of these two systems, which is essential for regulating immune function and mobilizing various physiological functions, and is an indispensable part of maintaining homeostasis. TRPV1, a cation channel belonging to the TRP (transient receptor potential) family, is an important chemosensitive and thermosensitive receptor, playing a vital role in sensing external environmental temperature and regulating body temperature. TRPV1 ion channels can be activated by various stimuli, such as capsaicin, heat (>43°C), acidic pH (<5.3), and endogenous substances. Furthermore, pro-inflammatory factors (such as histamine, prostaglandins, and ATP) can bind to G protein-coupled receptors (GPCRs), thereby triggering TRPV1 ion channel sensitization through phospholipase C (PLC) and protein kinase A (PKA)-dependent phosphorylation pathways. TRPV1+ neurons are widely distributed in the skin and many visceral tissues; the cell bodies of TRPV1+ neurons are located in the trigeminal ganglion, dorsal root ganglion, and jugular-nodal ganglion. TRPV1+ sensory neurons are responsible for transmitting noxious or thermal information from the body surface or internal tissues to the corresponding brain regions, thereby perceiving external environmental stimuli and the physiological state of internal tissues. Summary of the Invention

[0004] The purpose of this invention is to provide a method and drug for stimulating TRPV1+ sensory nerves to regulate the autonomic nervous system and organ function, treating visceral dysfunction, regulating hormone and catecholamine secretion, and preventing and treating inflammatory diseases caused by pro-inflammatory cytokines or pro-inflammatory cytokine cascades. The technical problems to be solved by this invention are not limited to the described technical topics; other technical topics not mentioned herein will be clearly understood by those skilled in the art through the following description.

[0005] To achieve the above objectives, the present invention first provides the use of a substance that stimulates TRPV1+ sensory nerves in the preparation of products having any of the following functions: A1) Prevention, treatment, or adjunctive treatment of inflammatory diseases; A2) Inhibit the expression of pro-inflammatory cytokines or inhibit the pro-inflammatory cytokine cascade; A3) Promotes the expression or production of anti-inflammatory cytokines; A4) Prevention, treatment, or adjunctive treatment of autonomic nervous system dysfunction; A5) Promotes catecholamine secretion; A6) Prevention, treatment, or adjuvant treatment of adrenal insufficiency; A7) Promotes the secretion of cortisol hormones by the adrenal glands.

[0006] In the above-mentioned applications, the substance that stimulates TRPV1+ sensory nerves can be a substance that activates TRPV1 ion channels through chemical stimulation, temperature stimulation, mechanical stimulation, electrical stimulation, ultrasound stimulation, or infrared stimulation.

[0007] In the above-mentioned uses, the substance that stimulates TRPV1+ sensory nerves may be a TRPV1 ion channel agonist.

[0008] In the above-mentioned applications, the TRPV1 ion channel agonist may be any of the following: B1) Reagents or drugs that activate TRPV1 ion channels; B2) Reagents or drugs that activate TRPV1+ sensory neurons; B3) Agents or drugs that promote the replication, transcription, translation, post-transcriptional modification and / or post-translational modification of the TRPV1 gene; B4) Reagents or drugs that increase or upregulate the content, activity, and / or function of TRPV1 protein.

[0009] Furthermore, the TRPV1 ion channel agonist can be a reagent or drug for overexpressing the TRPV1 gene (such as a recombinant vector for overexpressing the TRPV1 gene). Overexpression can be achieved through regulation at the gene level (such as gene replication, transcription, translation, post-transcriptional modification, and / or post-translational modification), or by promoting or increasing the content, activity, and / or function of the target protein at the protein level. There are no particular limitations on the means of overexpression, and many methods for achieving overexpression are well known to those skilled in the art. For example, the nucleic acid molecule to be overexpressed or the nucleic acid molecule encoding the TRPV1 protein can be placed under the control of a strong promoter; the copy number of one or more genes encoding the TRPV1 protein described in this invention can be increased; or the strength of the ribosome binding site or Kozak sequence can be increased, mRNA stability can be improved, codon usage can be altered, etc.

[0010] Furthermore, the TRPV1 ion channel agonists include, but are not limited to, the following chemicals: capsaicin, capsiate, capsaicinoid, deuterated capsaicin (capsaicin-d3), dihydrocapsaicin, dihydrocapsaicin ester, gingerol, allicin, allin, N-arachidonyldopamine (NADA), (R)-methanandamide (AM-356), evodiamine, cannabinoids, cannabidiol, vocacapsaicin (CA-008), and vocacapsaicin hydrochloride. hydrochloride, N-oleoyldopamine (OLDA), hydroxy-α-sanshool, nonivamide, paradol, protokylol, eugenol, camphor, clotrimazole, arvanil (N-arachidonoylvanillamine), anandamide, 2-aminoethoxydiphenyl borate (2APB), resiniferatoxin, phorbol 12-phenylacetate 13-acetate 20-homovanillate (PPAHV), olvanil, 6'-iodoresiniferatoxin, 6'-IRTX), C18 N-acylethanolamines, lipoxygenase derivatives, 12-hydroperoxyeicosatetraenoic acid, inhibitor cysteineknot peptides, vanillotoxins, piperine, N-[2-(3,[4-Dimethylbenzyl)-3-(neovaleroxy)propyl]-2-[4-(2-aminoethoxy)-3-methoxyphenyl]acetamide (MSK195), N-[2-(3,4-dimethylbenzyl)-3-(neovaleroxy)propyl]-N'-(4-hydroxy-3-methoxybenzyl)thiourea (JYL 79), 2-aminoethoxydiphenyl borate (2-aminoethoxydiphenyl) borate), zingerone, 10-shogaol, oleylgingerol, oleylshogaol, N-(4-tert-butylbenzyl)-N'-(4-hydroxy-3-methoxybenzyl)thiourea (SU200), amylocaine, articaine, benzocaine, bupivacaine, carbocaine, carticaine, chloroprocaine, cyclomethycaine, dibucaine (cinchocaine), dimethocaine (… larocaine, eticaine, hexylcaine, levobupivacaine, lidocaine, mepivacaine, meprylcaine, oracaine, metabutoxycaine, piperocaine, prilocaine, procaineNovacaine, proparacaine, propoxycaine, risocaine, ropivacaine, tetracaine (amethocaine), trimecaine, diallyl sulfides, alkylamides derived from sanshool, oxidized linoleic metabolites, black pepper compounds, tinyatoxin, AM404, MDR-652, MSP-3, OMDM-5, OMDM-6, CB1 / 2 agonist 4, SA13353.

[0011] Agonists of other receptors or ion channels expressed by TRPV1+ neurons can also activate TRPV1+ neurons, such as TRPA1 ion channels and agonists of some cytokine receptors.

[0012] The TRPA1 ion channel agonists include, but are not limited to, the following chemicals: allyl isothiocyanate (AITC), mustard oil, icilin, allicin, cannabidiol, gingerol, optovin, polygodial, resveratrol, diallyl disulfide, cinnamaldehyde, cinnamon oil, imiquimod, moringin, voacangine, wintergreen oil, clove oil, acrolein, hydroxy-alpha-sanshool, methyl syringate, and deuterated derivatives of methyl syringate. syringate-d6), 4-(Phenyldiazenyl)benzoic acid, (E)-4-Oxo-2-nonenal, 4-ONE, 2-aminoethoxydiphenyl borate, 4-hydroxynonenal, methyl p-hydroxybenzoate, 3'-carbamoylbiphenyl-3-yl Cyclohexylcarbamate (URB597), amylocaine, articaine, benzocaine, bupivacaine, carbocaine, carticaine, chloroprocaine, cyclomethycaine, dibucaine (cinchocaine), dimethocaine (larocaine), etidocaine, hexylcaine, levobupivacaine, lidocaine, mepivacaine.Meprylcaine (oracaine), metabutoxycaine, pipecaine, prilocaine, procaine (novacaine), proparacaine, propoxycaine, risocaine, ropivacaine, tetracaine (amethocaine), trimecaine, fenarnate, nicotine, diallyl sulfides nifedipine, nimodipine, nicardipine, nitrendipine, L-type calcium channel agonist BayK8644 BayK8644), hepoxilins A3 and B3, 12S-Hydroperoxy-5Z,8Z,10E, 14Z-eicosatetraenoic acid, 4,5-epoxyeicosatrienoic acid, supercimlamaldehyde, microRNA-711, microRNA-let-7b, ASP7663, AS1269574, PF-4840154, JT010.

[0013] In the above-mentioned applications, the TRPV1 ion channel agonist may be capsaicin or capsaicin-like substances.

[0014] The capsaicin-like compounds include capsaicin analogs and their derivatives, which are known to those skilled in the art and are commercially available.

[0015] Furthermore, the capsaicinoids include, but are not limited to, norcapsaicin, high capsaicin, dihydrocapsaicin, nordihydrocapsaicin (nordihydrocapsaicin), highdihydrocapsaicin, nonivamide, octanoylvanillamide, decanoylvanillamide, and zucapsaicin (cis-capsaicin, Civamide).

[0016] This invention also provides the use of the TRPV1 ion channel as a target in any of the following: C1) Use in the preparation of products for the prevention, treatment or adjunctive treatment of inflammatory diseases; C2) Use in the preparation of products that inhibit the expression of pro-inflammatory cytokines or inhibit the cascade reaction of pro-inflammatory cytokines; C3) Use in the preparation of products that promote the expression or production of anti-inflammatory cytokines; C4) Use in the preparation of products for the prevention, treatment or adjunctive treatment of autonomic nervous system dysfunction; C5) Use in the preparation of products that promote catecholamine secretion; C6) Use in the preparation of products for the prevention, treatment or adjunctive treatment of adrenal insufficiency; C7) Use in the preparation of products that promote the secretion of cortisol hormones by the adrenal glands; C8) Use in the development, design, or screening of drug candidates and / or treatments for the treatment and / or prevention of inflammatory diseases caused by pro-inflammatory cytokines or pro-inflammatory cytokine cascades; C9) Use in the development, design or screening of candidate drugs and / or treatments for the treatment and / or prevention of autonomic dysfunction disorders and diseases caused by insufficient catecholamine secretion; C10) Use in the development, design, or screening of candidate drugs and / or treatments for the treatment and / or prevention of diseases of adrenal insufficiency.

[0017] The present invention also provides pharmaceutical compositions comprising any of the TRPV1 ion channel agonists and / or TRPA1 ion channel agonists described herein, and one or more pharmaceutically acceptable carriers.

[0018] Furthermore, the pharmaceutical composition has at least one of the following uses: D1) Prevention, treatment, or adjunctive treatment of inflammatory diseases; D2) Inhibit the expression of pro-inflammatory cytokines or inhibit the pro-inflammatory cytokine cascade; D3) Promotes the expression or production of anti-inflammatory cytokines; D4) Prevention, treatment, or adjunctive treatment of autonomic nervous system dysfunction; D5) Promotes catecholamine secretion; D6) Prevention, treatment, or adjuvant treatment of adrenal insufficiency; D7) Promotes the secretion of cortisol hormones by the adrenal glands.

[0019] The pharmaceutically acceptable carrier is selected from diluents, excipients, fillers, binders, wetting agents, disintegrants, preservatives, stabilizers, absorption enhancers, adsorbents, surfactants, lubricants, atomizing agents, suspending agents, plasticizers, and dispersants.

[0020] In the above-mentioned uses, the pro-inflammatory cytokines may be TNFα, IL6, IL1a, IL1β, IL1f9, IL1r1, IL1rn, IL2, IL12b, IL13, IL15, IL16, IL20rb, IL27, TNFSF15, TNFSF18, TNFRSF12a, TNFRSF19, CD40, CSF2, CSF3, LIF, TIMP1, TSLP, FAS, OSMR, CCL1, CCL2, CCL3, CCL4, CCL5, CCL7, CCL11, CCL12, CCL1 7. CCL19, CCL20, CCL22, CCL27, CCR6, CXCL1, CXCL5, CXCL10, CXCL11, CX3CL1, XCL1, IFNa1, IFNa2, IFNa4, IFNa5, IFNa9, IFNab, IFNb1, IFNg, IFIH1, IFIT1, IFIT1bl1, IFIT2, IFIT3, IFIT3b, IFI27l2b, IFI47, IFI204, IFI205, IFI207, IFI211, IRF4 and / or IRF6.

[0021] In the above-mentioned uses, the anti-inflammatory cytokine may be IL10.

[0022] The corticosteroids mentioned in this article may refer to corticosterone.

[0023] The catecholamines mentioned in this article may be dopamine, norepinephrine, and / or epinephrine.

[0024] The present invention also provides a method for screening candidate drugs for the treatment and / or prevention of inflammatory diseases caused by pro-inflammatory cytokines or pro-inflammatory cytokine cascades, the method comprising: screening drugs to be screened using TRPV1 ion channels as targets, and selecting drugs that can activate TRPV1 ion channels or upregulate TRPV1 gene and / or TRPV1 protein levels as the candidate drugs.

[0025] The products described in this article may include reagents, formulations, drugs, pharmaceutical compositions, therapeutic kits, etc.

[0026] The present invention also provides the use of a substance that stimulates TRPV1+ sensory nerves in any of the following: E1) Prevention, treatment, or adjunctive treatment of inflammatory diseases; E2) Inhibits the expression of pro-inflammatory cytokines or inhibits the pro-inflammatory cytokine cascade; E3) promotes the expression or production of anti-inflammatory cytokines; E4) Prevention, treatment, or adjunctive treatment of autonomic nervous system dysfunction; E5) promotes catecholamine secretion; E6) Prevention, treatment, or adjuvant treatment of adrenal insufficiency; E7) promotes the secretion of cortisol hormones by the adrenal glands.

[0027] The present invention also provides a method for preventing or treating a disease, the method comprising stimulating the TRPV1+ sensory nerves of a subject or patient.

[0028] In the above methods, the diseases include inflammatory diseases, autonomic nervous system dysfunction diseases, or adrenal insufficiency diseases.

[0029] The above methods include administering to a subject or patient the TRPV1 ion channel agonist and / or TRPA1 ion channel agonist described herein, or the pharmaceutical composition described herein.

[0030] The patients described in this article may be those suffering from inflammatory diseases, autonomic nervous system dysfunction, or adrenal insufficiency.

[0031] The above methods include administration via intramuscular injection, intramuscular injection, subcutaneous injection, intradermal injection, transdermal injection, intravenous injection, arterial injection, intraperitoneal injection, intraperitoneal injection, microneedle injection, mucosal administration, oral administration, oral / nasal spray, nebulized inhalation, skin application, implantation, and external delivery via a portable device.

[0032] In the above method, the application sites include the abdominal cavity, the whole body skin, subcutaneous tissue, muscles, neck and back, waist and back, and abdomen.

[0033] Furthermore, the amount administered may be a therapeutically effective amount. A therapeutically effective amount may refer to the dosage of the drug used to (i) treat or prevent a specific disease, condition, or disorder; (ii) alleviate, improve, or eliminate one or more symptoms of a specific disease, condition, or disorder; or (iii) prevent or delay the onset of one or more symptoms of a specific disease, condition, or disorder described herein. A therapeutically effective amount can be determined by testing in a known in vitro or in vivo (e.g., animal model) system.

[0034] In the above methods, the stimulation includes chemical stimulation, temperature stimulation, mechanical stimulation, electrical stimulation, electrical signal stimulation, ultrasound stimulation, and infrared stimulation.

[0035] The temperature stimulation may include hot compresses, heat therapy, etc. The temperature stimulation may be a heat stimulation of 43℃, 44℃, 45℃, 46℃, 47℃, 48℃, 49℃ or 50℃.

[0036] The present invention also provides a method for inhibiting the expression of pro-inflammatory cytokines, the method comprising stimulating TRPV1+ sensory nerves in a subject or patient.

[0037] Further, the pro-inflammatory cytokines are selected from TNFα, IL6, IL1a, IL1β, IL1f9, IL1r1, IL1rn, IL2, IL12b, IL13, IL15, IL16, IL20rb, IL27, TNFSF15, TNFSF18, TNFRSF12a, TNFRSF19, CD40, CSF2, CSF3, LIF, TIMP1, TSLP, FAS, OSMR, CCL1, CCL2, CCL3, CCL4, CCL5, CCL7, CCL11, CCL12, and CCL... 17, CCL19, CCL20, CCL22, CCL27, CCR6, CXCL1, CXCL5, CXCL10, CXCL11, CX3CL1, XCL1, IFNa1, IFNa2, IFNa4, IFNa5, IFNa9, IFNab, IFNb1, IFNg, IFIH1, IFIT1, IFIT1bl1, IFIT2, IFIT3, IFIT3b, IFI27l2b, IFI47, IFI204, IFI205, IFI207, IFI211, IRF4 and IRF6.

[0038] The present invention also provides a method for promoting the expression of anti-inflammatory cytokines, the method comprising stimulating TRPV1+ sensory nerves in a subject or patient.

[0039] Furthermore, the anti-inflammatory cytokine is IL10.

[0040] The stimulation of TRPV1+ sensory nerves in subjects or patients described in this article can be achieved by activating TRPV1 ion channels or TRPV1+ sensory neurons through chemical stimulation, temperature stimulation, mechanical stimulation, electrical stimulation, electrical signal stimulation, ultrasound stimulation, or infrared stimulation.

[0041] The term "TRPV1+ sensory nerve" as used in this article can refer to sensory nerves or sensory neurons that express TRPV1, i.e., TRPV1-positive sensory nerve fibers or TRPV1-positive sensory neurons. The TRPV1+ sensory nerves include systemic TRPV1+ peripheral somatic sensory nerves, TRPV1+ internal tissue somatic sensory nerves, and TRPV1+ vagal sensory nerves.

[0042] The term "treatment" as used in this article includes both preventative and therapeutic treatment. "Preventative treatment" refers to preventing, suppressing, or reducing the occurrence of symptoms before they appear. "Therapeutic treatment" refers to treating subjects who already have the relevant disease.

[0043] Stimulation of TRPV1+ sensory nerves is effective in treating inflammatory diseases and regulating visceral function in body sites including, but not limited to, the following: abdominal cavity, skin, subcutaneous tissue, and muscles, especially the neck, back, lower back, and abdomen. Suitable routes of administration include: intradermal, transdermal, subcutaneous, intramuscular, intraperitoneal injections and oral administration of TRPV1+ neurostimulants; application of creams or gels to the skin; sprays; implants or external devices; and electrical stimulation, heat therapy, electrical stimulation, ultrasound stimulation, and infrared stimulation.

[0044] The term “inflammatory disease” is used in the broadest sense in this article, encompassing all diseases and pathological conditions related to systemic or local inflammatory responses, including acute and chronic inflammatory diseases.

[0045] Non-limiting examples of conditions for which this invention can be applied include autonomic nervous system (vegetative nervous system) disorders, adrenal insufficiency, cortisol hormone secretion disorders, catecholamine secretion disorders, sepsis, septicemia, septic shock, endotoxin shock, cytokine-induced toxicity, acute severe inflammation, chronic refractory inflammation, organ ischemia-reperfusion injury, organ necrosis, multiple organ injury syndrome, antigen-antibody complex-mediated diseases, immune responses associated with cytokine and T lymphocyte-mediated acute and delayed hypersensitivity reactions, allergic diseases, myocardial or other tissue reperfusion injury, cardiovascular ischemia, ischemic-reperfusion disorder, meningitis, encephalitis, Rasmussen encephalitis, limbic and / or brainstem encephalitis, cerebral infarction, cerebral embolism, cerebral ischemia, and acute suppuration. Meningitis or other inflammatory diseases of the central nervous system, neuritis, neuralgia, hyperalgesia, polyradiculitis, spinal cord injury, paralysis, rheumatoid spondylitis, inflammatory diseases of the central nervous system, allergic neuritis, systemic lupus erythematosus, arthritis, acute arthritis, rheumatoid arthritis, chronic rheumatoid arthritis, gouty arthritis, acute gouty arthritis, chronic inflammatory arthritis, degenerative arthritis, infectious arthritis, Lyme arthritis, proliferative arthritis, psoriatic arthritis, spondyloarthritis and juvenile-onset rheumatoid arthritis, osteoarthritis, chronic progressive arthritis, deforming arthritis, seronegative spondyloarthritis, chronic primary polyarthritis, reactive arthritis and ankylosing spondylitis, rheumatic diseases. Sclerosis, Systemic sclerosis, Multiple sclerosis, Spinal optical multiple sclerosis, Primary progressive multiple sclerosis, Relapsing-remitting multiple sclerosis, Progressive systemic sclerosis, Atherosclerosis, Arteriosclerosis, Diffuse sclerosis and ataxia-associated sclerosis, Autoimmune neutropenia, Pancytopenia, Leukopenia, Diseases involving leukocyte exudation, Eosinophil-related diseases such as eosinophilia, Infiltrative eosinophilia, Eosinophilia-myalgia syndrome, Ruffler syndrome, Chronic eosinophilic pneumonia, Tropical eosinophilic pneumonia, Granulocyte transfusion-related syndrome, Stiff man or stiff man syndrome, Encephalomyelitis, Allergic encephalomyelitis, Laboratory Allergic encephalomyelitis, transverse myelitis, myasthenia gravis, thymoma-associated myasthenia gravis, cerebellar degeneration, neuromuscular rigidity, optoclonus or optoclonus myoclonus syndrome and sensory neuropathy, multifocal motor neuropathy, neuromyelitis optica, polyneuropathy, chronic neuropathy, IgM polyneuropathy, IgM-mediated neuropathy, peripheral neuropathy, ion channelopathies such as epilepsy, migraine, periodic paralysis and central nervous system channelopathies, autism, Alzheimer's disease, demyelinating diseases, Alzheimer's disease, diseases involving T-cell infiltration, leukocyte adhesion deficiency, diseases involving leukocyte exudation, allogeneic transplant rejection, graft-versus-host disease, respiratory distress syndrome, influenza, influenza virus infection, coronavirus infection.Respiratory syncytial virus infection, adenovirus infection, parainfluenza virus infection, hepatitis B virus infection, hepatitis C virus infection, metapneumovirus infection, cytomegalovirus infection, smallpox infection, herpesvirus infection, disseminated bacteremia, dengue fever, candidiasis, human immunodeficiency virus (HIV) infection, echovirus infection, parvovirus infection, rubella virus infection, post-vaccination syndrome, congenital rubella infection, Epstein-Barr virus infection, mumps, sequelae of streptococcal nephritis, malaria, filariasis, amoebiasis, leprosy, leishmaniasis, trypanosomiasis, schistosomiasis, ascariasis, aspergillosis, Samptothenia syndrome, Kaplan syndrome, parasitic diseases such as leishmaniasis, burns, vesicular cysts, asthma, pharyngitis, thyroiditis, and so on. Autoimmune thyroiditis, chronic thyroiditis, subacute thyroiditis, Hashimoto's disease, primary hypothyroidism, hypoparathyroidism, autoimmune thyroid disease, idiopathic hypothyroidism, hyperthyroidism, allergic and atopic rhinitis, bronchitis, bronchiolitis, pneumonia, lymphoid interstitial pneumonia, obliterative bronchiolitis (non-transplant), emphysema, pneumoconiosis, pulmonary cirrhosis, alveolitis, bird laryngotracheitis, chronic hypersensitivity pneumonia, farmer's lung, alveolitis, bronchitis, chronic obstructive airway disease, silicosis, pulmonary sclerosis, interstitial lung disease, transfusion reaction, endocardial fibrosis, diffuse interstitial pulmonary fibrosis, interstitial pulmonary fibrosis, idiopathic pulmonary fibrosis, pleurisy, autoimmune myocarditis Pericarditis, myocarditis, dilated cardiomyopathy, endocarditis, myocardial ischemia, arrhythmia, post-myocardial infarction syndrome, post-cardiotomy syndrome, cardiomyopathy, valvulitis, thromboangiitis obliterans, choroiditis, arteriosclerotic diseases, idiopathic nephrotic syndrome, minimal change disease, regional ileitis, diabetic aortic disease, pyelonephritis, intra-arterial hyperplasia, peptic ulcer, pancreatitis, acquired splenic atrophy, periarteritis nodosa, vasculitis, arteritis, thrombophlebitis, vasculitis, giant cell (Gaumann's) arteritis, mid-segment vasculitis, allergic granulomatous vasculitis, benign lymphocytic vasculitis, Allport syndrome, polyarteritis nodosa, microscopic polyarteritis, central nervous system vasculitis, necrotizing, cutaneous, hypersensitivity Vasculitis, systemic necrotizing vasculitis, ANCA-associated vasculitis, DeQuailwan's thyroiditis, thyrotoxicosis, allergic granulomatous vasculitis or syndrome, temporal arteritis, celiac disease, appendicitis, gastric atrophy, autoimmune atrophic gastritis, peptic ulcer, gastric ulcer, duodenal ulcer, peritonitis, pancreatitis, ulcerative colitis, pseudomembranous colitis, acute colitis, ischemic colitis, cholangitis, cholecystitis, nephrotic syndrome, pancreatitis, hepatitis, Crohn's disease, enteritis, inflammatory bowel disease, irritable bowel syndrome, Crohn's disease, colitis, ulcerative colitis, collagenous colitis, polypoid colitis, necrotizing enterocolitis, transmural colitis, autoimmune enteropathic syndrome, autoimmune hepatitis, chronic hepatitis.Lupus-like hepatitis, giant cell hepatitis, chronic active hepatitis or autoimmune chronic active hepatitis, autoimmune liver disease, non-alcoholic fatty liver disease, Guillain-Barré syndrome, primary biliary cirrhosis, autoimmune orchitis and oophoritis, epididymitis, vaginitis, prostatitis, urethritis, sinusitis, dermatitis, dermatomyositis, primary sclerosing cholangitis, superficial scleritis, total or partial uveitis, iritis, choroiditis, uveitis, anterior uveitis, acute anterior uveitis, granulomatous uveitis, non-granulomatous uveitis, superantigen uveitis, posterior uveitis, autoimmune uveitis, pulmonary hemorrhage nephritis syndrome, glomerulonephritis, idiopathic membranous glomerulonephritis or idiopathic membranous nephropathy, membranous Or membranous proliferative nephritis, rapidly progressive glomerulonephritis, Burger's disease (IgA nephropathy), idiopathic IgA nephropathy, immune complex nephritis, antibody-mediated nephritis, adrenalitis, diabetic nephropathy, glomerulonephritis, nephropathy, granulomatous orchitis, leukopenia, juvenile-onset type I diabetes, childhood insulin-dependent diabetes, adult-onset type II diabetes, autoimmune diabetes, idiopathic diabetes insipidus, tuberculosis, sarcoidosis, granulomatous disease, lymphomatoid granulomatosis, Wegener's granulomatosis, agranulocytosis, polymyalgia rheumatica, Kawasaki disease, aplastic anemia, autoimmune aplastic anemia, Coombs-positive anemia, congenital pure red cell aplasia. Aplastic anemia, hemolytic anemia, or immune hemolytic anemia, including autoimmune hemolytic anemia, pernicious anemia, Addison's disease, pure red cell anemia or aplastic anemia, factor VIII deficiency, hemophilia, autoimmune blood disorders, hypotensive response, vascular dysfunction, vasodilation, antiglomerular basement membrane disease, antiphospholipid antibody syndrome, Behcet's disease or Behcet's disease, Kassman syndrome, Raynaud's syndrome, Sjögren's syndrome, Stevens-Johnson syndrome, pemphigus, selective pemphigus vulgaris, foliaceous pemphigus, mucosal pemphigus, erythematous pemphigus, autoimmune multiple endocrine disorders, Leter's disease or syndrome, thrombocytopenia, thrombotic thrombocytopenic purpura, idiopathic thrombocytopenic purpura. Autoimmune hemolysis, Burkholderia, cryoglobulinemia, polyglandular syndrome, paraneoplastic syndromes, including neurological paraneoplastic syndromes such as Lambert-Eton myasthenia gravis or Eaton-Lambert syndrome, Sheehan's syndrome, linear IgA dermatitis, celiac disease, stomatitis, refractory aphthous stomatitis, idiopathic aphthous stomatitis, cryoglobulinemia, amyotrophic lateral sclerosis, coronary artery disease, ataxia-telangiectasia, autoimmune ear diseases such as autoimmune inner ear disease, autoimmune hearing loss, optoclonus-myoclonus syndrome, polychondritis, pulmonary alveolar proteinosis, amyloidosis, scleritis, noncancerous lymphocytosis, essential lymphocytosis, including monoclonal B-cell lymphocytosis.Selective benign monoclonal gammopathy or monoclonal gammopathy of undetermined significance, paraneoplastic syndromes, muscle diseases, inflammatory myopathy, fibromyalgia, multiple endocrine failure, Schmidt syndrome, CREST syndrome, male and female autoimmune infertility, mixed connective tissue diseases, Chagas disease, rheumatic fever, recurrent miscarriage, erythema multiforme, Cushing's syndrome, cystic fibrosis, focal segmental hearing loss, sudden hearing loss, blindness, keratoconjunctivitis sicca, epidemic keratoconjunctivitis, endocrine eye diseases, uveitis, inflammatory diseases of the eye and orbit, diabetic retinopathy, choroidal retina. Inflammation, scleritis, episcleritis, endophthalmitis, retinal autoimmune disease, erythema elevata, fetal erythrocytosis, eosinophilic fasciitis, purulent or non-purulent sinusitis, acute or chronic sinusitis, Shulman syndrome, Felty syndrome, cyclitis, endocrine ophthalmitis, hypersensitivity endophthalmitis, sympathetic ophthalmitis, allergic purpura, Evan syndrome, autoimmune gonadal insufficiency, Sidnam's chorea, peripheral tabes dorsalis, polymyalgia giant cells, aphthous stomatitis, aphthous stomatitis, azoospermia, infertility caused by antisperm antibodies, Dupuitren's contracture, erythema nodosum leprosy, idiopathic facial paralysis, chronic fatigue syndrome Symptoms, rheumatic fever, Harman-Ritchie disease, sensorineural hearing loss, paroxysmal hemoglobinuria, hypogonadism, leukopenia, infectious mononucleosis, primary idiopathic myxedema, skin diseases with acute inflammatory components, pyoderma gangrene, non-malignant thymoma, vitiligo, SCID and Barr virus-related diseases, acquired immunodeficiency syndrome (AIDS), toxic shock syndrome, food poisoning, pyoderma gangrene, erythema nodosum, antiglomerular basement membrane disease, allergic neuritis, autoimmune polyendocrine disorders, primary myxedema, sympathetic ophthalmitis, mixed connective tissue disease, multiple endocrine disorders Endocrine dysfunction, autoimmune polygonococcal syndrome type I, adult-onset idiopathic hypoparathyroidism, acquired epidermolysis bullosa, hemochromatosis, nephrotic syndrome, primary sclerosing cholangitis, ethmoid sinus, frontal sinus, maxillary sinus, or sphenoid sinusitis, bronchopneumonia with aspergillosis, aspergilloma or granuloma containing eosinophils, endometriosis, polyendocrine autoimmune diseases, sclerosing cholangitis, chronic mucocutaneous candidiasis, Bruton's syndrome, transient hypogammaglobulinemia in infants, Wescott-Aldridge syndrome, collagen-related autoimmune diseases, and allergic hypersensitivity disorders.

[0046] This invention establishes a systemic inflammation mouse model by injecting LPS (lipopolysaccharide, a component of the cell wall of Gram-negative bacteria). It was found that stimulating TRPV1+ sensory nerves in different parts of the mouse body can reduce the expression of various cytokines (such as TNF-α, IL-6, and IL-1β) to varying degrees, with subcutaneous injection of capsaicin in the neck and back being the most effective anti-inflammatory method. Figure 1The anti-inflammatory effect of stimulating TRPV1+ sensory nerves depends on the capsaicin dosage. Transdermal stimulation of TRPV1+ sensory nerves with capsaicin cream effectively inhibits cytokine expression, indicating that direct stimulation of TRPV1+ sensory nerves in the skin is effective in reducing inflammation. Figure 2 Luminex protein multifactor analysis was used to detect the expression of 31 cytokines. It was found that stimulation of TRPV1+ sensory nerves inhibited the expression of numerous pro-inflammatory cytokines and chemokines, while increasing the expression of anti-inflammatory cytokines (IL-10). trpv1 Capsaicin pretreatment in knockout mice did not produce an anti-inflammatory effect; therefore, capsaicin exerts its anti-inflammatory effect by specifically stimulating TRPV1+ sensory nerves. Figure 3 Thermal stimulation of TRPV1+ cutaneous sensory nerves at approximately 50°C significantly inhibited the expression of numerous pro-inflammatory cytokines, chemokines, and interferons, while enhancing the expression of the anti-inflammatory cytokine IL10. In contrast, cold stimulation at approximately 0°C only inhibited the expression of TNFα and CCL3. Therefore, thermal stimulation of TRPV1+ cutaneous sensory nerves, similar to moxibustion, can effectively reduce inflammation. Figure 4 In lethal systemic inflammation, stimulation of TRPV1+ sensory nerves significantly improved the survival rate and physiological state of inflammatory mice. Figure 5 Through pre-splenectomy and reserveine drug pretreatment experiments, this study reveals that stimulating TRPV1+ sensory nerves regulates spleen function and thus reduces inflammation by activating the sympathetic nervous system. Figure 6 Transcriptome sequencing of spleen gene expression revealed that stimulation of TRPV1+ sensory nerves significantly affected spleen gene expression, particularly inhibiting the expression of pro-inflammatory genes, regardless of whether the condition was inflammatory or normal. Functional analysis showed that differentially expressed genes were mainly enriched in inflammation-related signaling pathways. Figure 7 By dynamically monitoring changes in serum catecholamine concentrations in mice, and through subphrenic vagal nerve transection surgery and hexamethonium bromide pretreatment, it was found that stimulation of TRPV1+ somatic sensation could activate the vagus nerve-adrenal axis and increase catecholamine production. trpv1 In knockout mice, injection of capsaicin did not promote the expression of the three catecholamines. Figure 8 In addition, stimulation of TRPV1+ sensory nerves can activate the hypothalamic-pituitary-adrenal reflex arc and promote the secretion of cortisol hormones. trpv1 In knockout mice, corticosterone expression did not increase in mice injected with capsaicin. Figure 9 ).

[0047] In summary, stimulating TRPV1+ peripheral sensory nerves can regulate the somatic sensory nerve-autonomic nervous system-immune system reflex arc, thereby synergistically inhibiting the production of pro-inflammatory cytokines. Simultaneously, it activates the vagus nerve-adrenal axis, increasing catecholamine production, and activates the hypothalamic-pituitary-adrenal reflex arc, promoting cortisol secretion. Therefore, sensory nerves not only perceive changes in the external environment and the state of internal tissues, but also regulate visceral function and the body's physiological state through sensory nerve-autonomic nerve reflex arcs, sensory nerve-hormonal reflex arcs, and neuroimmune interactions. This invention suggests that temperature stimulation, chemical stimulation, mechanical stimulation, tissue damage, or pathogen infection of the body surface activate the peripheral sensory nervous system, thereby momentarily or continuously interfering with visceral function and disrupting homeostasis. In severe cases, this can lead to physical discomfort and disease. Accordingly, corresponding preventive measures can be taken to prevent physiological imbalances.

[0048] This invention is based on the inventors' discovery that stimulating sensory nerves can regulate visceral function and the body's physiological state. This mechanism can be used to regulate the body's homeostasis, thereby preventing and treating diseases. This invention utilizes this important physiological mechanism to disclose methods and drugs for treating visceral dysfunction by activating TRPV1+ sensory nerves to regulate the autonomic nervous system and organ function, regulating hormone and catecholamine secretion, effectively reducing the production of pro-inflammatory cytokines and thus inhibiting inflammation, regulating the body's physiological state, and preventing and treating inflammatory diseases caused by pro-inflammatory cytokines or their cascade reactions.

[0049] Therefore, this invention proposes novel uses for substances that stimulate TRPV1+ sensory nerves (such as substances that activate TRPV1 ion channels through chemical stimulation, temperature stimulation, mechanical stimulation, electrical stimulation, ultrasound stimulation, or infrared stimulation) in the prevention, treatment, or adjuvant treatment of inflammatory diseases, autonomic nervous system disorders, and adrenal insufficiency. Previously, the art had not conceived of using TRPV1 ion channel agonists as targets for the prevention or treatment of inflammatory diseases, autonomic nervous system disorders, and adrenal insufficiency. Unexpectedly, the inventors of this application have experimentally demonstrated that TRPV1 ion channel agonists (such as capsaicin or heat-stimulating substances) can significantly and effectively inhibit the expression of numerous pro-inflammatory cytokines and significantly increase the expression of the anti-inflammatory cytokine IL10, not only significantly improving the physiological state of inflamed mice but also significantly increasing their survival rate. Simultaneously, TRPV1 ion channel agonists can also regulate spleen function, significantly inhibit the expression of pro-inflammatory genes in the spleen, increase the production of catecholamines (dopamine, norepinephrine, and epinephrine), and promote the secretion of cortisol hormones. This invention can also be used to screen candidate drugs for the treatment and / or prevention of inflammatory diseases, autonomic nervous system dysfunction, and adrenal insufficiency caused by pro-inflammatory cytokines or pro-inflammatory cytokine cascades, and to develop new treatment methods and drugs. This invention has broad clinical application value for the prevention and treatment of related diseases. Attached Figure Description

[0050] Figure 1 A schematic diagram illustrating the anti-inflammatory effects of stimulating TRPV1+ sensory nerves in different parts of a mouse body.

[0051] Figure 2 A schematic diagram showing that the anti-inflammatory effect of stimulating TRPV1+ sensory nerves depends on the capsaicin dose, and that stimulating TRPV1+ cutaneous sensory nerves can effectively inhibit cytokine expression.

[0052] Figure 3 A schematic diagram showing how stimulating TRPV1+ sensory nerves inhibits the expression of numerous cytokines and chemokines, and how this anti-inflammatory response depends on TRPV1+ sensory nerves.

[0053] Figure 4 This is a schematic diagram illustrating how thermal stimulation of the TRPV1 ion channel can effectively inhibit the expression of numerous pro-inflammatory cytokines and chemokines.

[0054] Figure 5 This diagram illustrates how stimulating TRPV1+ sensory nerves can improve the survival rate and physiological state of inflammatory mice.

[0055] Figure 6 This diagram illustrates how stimulating TRPV1+ sensory nerves can regulate spleen function and thus reduce inflammation by activating the sympathetic nervous system.

[0056] Figure 7 This is a schematic diagram showing that stimulating TRPV1+ sensory nerves significantly inhibits the expression of pro-inflammatory genes in the spleen, as demonstrated by spleen transcriptome sequencing.

[0057] Figure 8 A schematic diagram illustrating how stimulating TRPV1+ sensory nerves can activate the vagus nerve-adrenal axis and increase catecholamine production.

[0058] Figure 9 A schematic diagram illustrating how stimulating TRPV1+ sensory nerves can activate the hypothalamic-pituitary-adrenal reflex arc and promote the secretion of cortisol hormones. Detailed Implementation

[0059] The present invention will now be described in further detail with reference to specific embodiments. The given embodiments are merely illustrative of the invention and not intended to limit its scope. The embodiments provided below can serve as a guide for further improvements by those skilled in the art and do not constitute a limitation on the invention in any way.

[0060] Unless otherwise specified, the experimental methods used in the following examples are conventional methods, performed according to the techniques or conditions described in the literature in this field or according to the product instructions. Unless otherwise specified, the materials and reagents used in the following examples are commercially available.

[0061] The biomaterials and reagents involved in the following examples are as follows: 1. Animals C57BL / 6J mice were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. Trpv1- / -(KOCMP-193034-Trpv1) mice (hereinafter referred to as...) trpv1 Knockout mice were purchased from Cyagen Biotech Ltd. All animals were housed in a standard barrier environment, with free access to standard mouse feed and water. All animal experiments were conducted in accordance with the experimental protocols approved by the Animal Ethics Committee of the Beijing Institute of Biotechnology.

[0062] 2. Biological reagents LPS (L2630), Reserpine (83580), and Hexamethonium bromide (H0879) were purchased from Sigma-Aldrich. Capsaicin (A3278) was purchased from Apexbio. Capsaicin cream (Shuxiao) was manufactured by Puhua Pharmaceutical. Parameter Corticosterone Assay kit (KGE009), IL-1β (MLB00C), and IL-6 (M6000B) ELISA kits were purchased from R&D Systems. TNFα (EK282 / 4-96) ELISA kits were purchased from Linko Biotech. Bio-Plex Pro Mouse Chemokine Panel 31-Plex (12009159) was purchased from Bio-Rad. 3-CAT ELISA kit (BA E-5600R) was purchased from LDN.

[0063] The solvents used in the following examples are all physiological saline.

[0064] Example 1: Comparison of anti-inflammatory effects of stimulating TRPV1+ sensory nerves in different parts of mice. A systemic inflammation mouse model was established by intraperitoneal injection of 8 mg / kg LPS (endotoxin) into 6-8 week old male C57BL / 6J mice. Thirty minutes before LPS injection, 4 mg / kg capsaicin (CAP, a TRPV1 ion channel agonist) was injected intraperitoneally, subcutaneously in the abdomen, neck, or back, or 0.25 mg / kg via the tail vein to establish the CAP-LPS experimental group. A control group consisting of a solvent (physiological saline) + LPS was also included. Blood samples were collected 1.5 hours and 6 hours after LPS injection, left at room temperature for 2 hours, and then centrifuged at 2000 g for 20 minutes to collect serum. The serum was stored at -80°C before use. The expression levels of serum cytokines TNFα, IL6, and IL1β were detected using TNFα, IL6, or IL1β ELISA kits according to the manufacturer's instructions.

[0065] The results are as follows Figure 1As shown, capsaicin treatment at different sites effectively inhibited TNFα expression. Subcutaneous injection of capsaicin into the neck and back inhibited IL6 expression at both 1.5 and 6 hours, while subcutaneous injection into the abdomen only suppressed IL6 production at 1.5 hours. Other injection sites had no effect on IL6 expression. IL1β expression was undetectable at 1.5 hours after LPS injection, but significantly increased at 6 hours. Only subcutaneous injection of capsaicin into the neck and back or lower back effectively inhibited IL1β production. Therefore, stimulation of TRPV1+ sensory nerves at different sites can reduce cytokine expression to varying degrees, with subcutaneous injection of capsaicin into the neck and back being the most effective anti-inflammatory method.

[0066] Example 2: Effect of capsaicin dosage on the anti-inflammatory effect of stimulating TRPV1+ sensory nerves Without LPS, blood samples were collected 1.5 and 6 hours after a subcutaneous injection of 4 mg / kg capsaicin into the back of the neck. Thirty minutes before LPS injection, mice were injected subcutaneously with 1 mg / kg or 4 mg / kg capsaicin, or capsaicin cream was applied to the skin of the back of the neck after shaving. Blood samples were collected 1.5 and 6 hours after LPS injection, incubated at room temperature for 2 hours, and then centrifuged at 2000 g for 20 minutes to collect serum. The serum was stored at -80°C before use. Serum cytokine expression levels were detected using either a TNFα or IL6 ELISA kit, following the manufacturer's instructions.

[0067] The results are as follows Figure 2 As shown, without LPS injection, 4 mg / kg capsaicin only produced basal levels of cytokine expression. Subcutaneous injection of 1 mg / kg capsaicin into the back of the neck resulted in a 55.3% decrease in TNFα expression, while 4 mg / kg capsaicin caused a 70.1% decrease. 1 mg / kg capsaicin led to a decrease in IL6 expression. 4 mg / kg capsaicin caused a 92% decrease in IL6 expression at 1.5 hours and a 72.4% decrease at 6 hours. Capsaicin cream effectively inhibited cytokine expression through transdermal stimulation of TRPV1+ cutaneous sensory nerves. This indicates that direct stimulation of TRPV1+ sensory nerves in the skin is effective in reducing inflammation. The above results show that the anti-inflammatory effect of stimulating TRPV1+ sensory nerves depends on the capsaicin dose. When the capsaicin dose is greater than or equal to 1 mg / kg, it can reduce the expression levels of cytokines TNFα and IL6. When the capsaicin dose is greater than or equal to 4 mg / kg, the inhibitory effect on cytokines is better, and transdermal stimulation of TRPV1+ cutaneous sensory nerves can effectively inhibit cytokine expression.

[0068] Example 3: Stimulation of TRPV1+ sensory nerves in the neck and back inhibits the expression of numerous cytokines and chemokines. Without LPS, blood was collected 1.5 hours after a subcutaneous injection of either a solvent (physiological saline) or 4 mg / kg capsaicin into the back of the neck. Thirty minutes before LPS injection, 4 mg / kg capsaicin was subcutaneously injected into the back of the neck of mice to establish the CAP-LPS experimental group, with a solvent + LPS control group also included. trpv1 In knockout mice, LPS was injected intraperitoneally 30 minutes after a subcutaneous injection of 4 mg / kg capsaicin into the neck and back, serving as the CAP-LPS experimental group. A solvent + LPS control group was also included. Blood samples were collected 1.5 hours after LPS injection, left at room temperature for 2 hours, and then centrifuged at 2000 g for 20 minutes to collect serum. The serum was stored at -80°C before use. Following the manufacturer's instructions, the expression of 31 cytokines was detected using the Bio-Plex Pro Mouse Chemokine Panel 31-Plex kit via Luminex multifactor protein analysis.

[0069] The results are as follows Figure 3 As shown, without LPS injection, subcutaneous injection of 4 mg / kg capsaicin only inhibited basal IL6 expression in mice. LPS injection led to an increase in the expression of 13 pro-inflammatory cytokines and chemokines, while pre-injection of capsaicin subcutaneously into the neck and back significantly reduced the expression of these 13 cytokines and chemokines and increased the expression of the anti-inflammatory cytokine IL10. trpv1 Capsaicin pretreatment in knockout mice did not produce an anti-inflammatory effect, therefore capsaicin exerts its anti-inflammatory effect by specifically stimulating TRPV1+ sensory nerves. These results indicate that stimulation of TRPV1+ sensory nerves in the neck and back can inhibit the expression of numerous cytokines and chemokines, and that this anti-inflammatory response is dependent on TRPV1+ sensory nerves.

[0070] Example 4: Thermal stimulation of TRPV1 ion channels in the neck and back After shaving the neck and back of C57BL / 6J mice, the skin was exposed to either approximately 50°C heat stimulation (using a 50°C hand warmer, similar to moxibustion) or 0°C cold stimulation (using an ice cube) for 15 minutes, followed by an intraperitoneal injection of LPS. Three groups were established: an untreated LPS control group, a 50°C-LPS experimental group, and a 0°C-LPS experimental group. Blood samples were collected 1.5 hours after LPS injection, left at room temperature for 2 hours, and then centrifuged at 2000 g for 20 minutes to collect serum. The serum was stored at -80°C before use. Following the manufacturer's instructions, the expression of 31 cytokines was detected using the Bio-Plex Pro Mouse Chemokine Panel 31-Plex kit and Luminex multifactor protein analysis.

[0071] The results are as follows Figure 4As shown, TRPV1 ion channels can be activated by temperatures >43°C. Thermal stimulation at approximately 50°C significantly inhibited the expression of numerous pro-inflammatory cytokines, chemokines, and interferons, while increasing the expression of the anti-inflammatory cytokine IL10. Cold stimulation at approximately 0°C only inhibited the expression of TNFα and CCL3. Therefore, heat stimulation of TRPV1+ cutaneous sensory nerves, similar to moxibustion, can effectively reduce inflammation. These results indicate that thermal stimulation of TRPV1 ion channels in the neck and back can effectively inhibit the expression of numerous pro-inflammatory cytokines and chemokines.

[0072] Example 5: Stimulation of TRPV1+ sensory nerves in the neck and back can improve the survival rate and physiological state of inflamed mice. A systemic inflammation mouse model was established by intraperitoneal injection of 8 mg / kg LPS into male mice aged 9-11 weeks. Thirty minutes before LPS injection, 4 mg / kg capsaicin was subcutaneously injected into the neck and back of C57BL / 6J mice to establish the CAP-LPS experimental group. n = 25), and a solvent + LPS control group was set up ( n = 19). 30 minutes before LPS injection, in trpv1 Knockout mice were subcutaneously injected with 4 mg / kg capsaicin in the back of their necks to establish the CAP-LPS experimental group. n = 17), and a solvent + LPS control group was set up ( n = 13). The physiological status of mice was assessed every 12 hours before and after LPS injection. Based on scores for body temperature, weight, coat color, and activity level, mice with inflammation were assessed to determine whether they met the death criteria and were promptly euthanized. Mice were considered dead if they met any of the following criteria: 1. Body temperature (body temperature <32℃); 2. Weight (weight loss of 20%); 3. Total clinical symptom score (a total score of 2 points or higher). Clinical symptoms included four categories, each scored in three levels (0, 1, 2): a. Eye discharge (excessive discharge causing eye closure: 0 points; excessive discharge: 1 point; no discharge: 2 points); b. Diarrhea (severe: 0 points; loose stools: 1 point; normal: 2 points); c. Coat color (dry, dull, and ungroomed fur: 0 points; dry and poorly groomed fur: 1 point; shiny fur: 2 points); d. Activity level (drowsy and only raising head with mild stimulation: 0 points; inactive and only moving less than two steps with mild stimulation: 1 point; normal: 2 points). A total score of 8 points across all four categories indicates a mouse in a normal physiological state. The mice were assessed to determine whether they met the criteria for early termination of the experiment and were promptly euthanized. The time of death was recorded. All experimental mice were euthanized on day 14 after drug administration, which was the termination point for the survival experiment.

[0073] The results are as follows Figure 5As shown, the survival rate of inflammatory mice in the control group was only 7%, while the survival rate of inflammatory mice injected with capsaicin in the experimental group was as high as 60%. We continued to observe the experimental mice for 2 weeks. All surviving mice regained their body weight after 1 week of experimentation and no further deaths occurred, indicating that stimulating TRPV1+ sensory nerves can continuously protect mice against inflammatory attacks. In addition, the body temperature changes of inflammatory mice injected with capsaicin in the experimental group were significantly better than those of control mice. trpv1 In the knockout mice, the experimental group injected with capsaicin trpv1 The survival rate and body temperature changes of the knockout inflammation mice were no different from those of the control mice.

[0074] Example 6: Stimulation of TRPV1+ sensory nerves regulates spleen function and thus reduces inflammation by activating the sympathetic nervous system. Splenectomy: Mice underwent surgery under isoflurane anesthesia. After opening the abdomen, the stomach was gently moved to expose the spleen. The blood vessels connecting to the spleen were ligated with nylon surgical sutures, and then the spleen was removed. The abdominal cavity was then sutured, and analgesics were injected subcutaneously. Experiments were conducted 3 days after surgery. Thirty minutes before LPS injection, 4 mg / kg capsaicin was subcutaneously injected into the neck and back of the splenectomized mice to establish the CAP-LPS experimental group, with a solvent + LPS control group.

[0075] Reserpine treatment experiment (Reserpine is a peripheral sympathetic inhibitor used to block peripheral sympathetic nerves; Martínez-Olivares, R., Villanueva, I., Racotta, R. & Piñón, M. Depletion and recovery of catecholamines in several organs of rats treated with reserpine.) Autonomic Neuroscience 128, 64-69 (2006). Mice were intraperitoneally injected with 10 mg / kg Reserpine, and subsequent experiments were conducted 24 hours later. Thirty minutes before LPS injection, mice treated with Reserpine were subcutaneously injected with 4 mg / kg capsaicin in the neck and back to establish the CAP-LPS experimental group, and a solvent + LPS control group was set up.

[0076] Blood samples were collected 1.5 hours after LPS injection, left at room temperature for 2 hours, and then centrifuged at 2000 g for 20 minutes to collect serum. The serum was stored at -80°C before use. Serum TNFα and IL6 expression levels were detected using either a TNFα or IL6 ELISA kit, following the manufacturer's instructions.

[0077] The results are as follows Figure 6As shown, capsaicin treatment after spleen removal did not induce an anti-inflammatory response. Reserpine inhibits the release of norepinephrine from peripheral sympathetic nerve endings, thereby suppressing the transmission of sympathetic nerve signals. After reserpine blocks peripheral sympathetic nerves, capsaicin treatment did not produce an anti-inflammatory effect.

[0078] Example 7: Stimulation of TRPV1+ sensory nerves significantly inhibited the expression of pro-inflammatory genes in the spleen. Without LPS, spleens were harvested 1 hour after a subcutaneous injection of either the solvent or 4 mg / kg capsaicin into the back of the neck. Thirty minutes before LPS injection, mice were subcutaneously injected with 4 mg / kg capsaicin into the back of the neck to establish the CAP-LPS experimental group, with a solvent + LPS control group. Spleens were harvested 1 hour after LPS injection. Beijing Biomarker Biotechnology Co., Ltd. provided transcriptome sequencing and related data analysis services.

[0079] The results are as follows Figure 7 As shown, stimulation of TRPV1+ sensory nerves significantly affects the expression of spleen genes, especially inhibiting the expression of pro-inflammatory genes (including Tnf, Il6, Ifng, etc.) under both inflammatory and normal physiological conditions. Functional analysis showed that differentially expressed genes were mainly enriched in inflammation-related signaling pathways.

[0080] Example 8: Stimulation of TRPV1+ somatic sensory nerves can activate the vagus nerve-adrenal axis and increase the production of catecholamines. In wild type and trpv1 Knockout mice were subcutaneously injected with 4 mg / kg capsaicin in the neck and back to establish an experimental group, and a solvent control group was set up. Blood samples were collected at different time points.

[0081] Subphrenic vagus nerve transection surgery: Mice underwent surgery under isoflurane anesthesia. After abdominal incision, the stomach was gently moved to expose the esophagus, and the two subphrenic vagus nerves were identified and transcribed. The abdominal cavity was then sutured, and analgesics were injected subcutaneously. Experiments were conducted 3 days after surgery. Mice with transcribed subphrenic vagus nerves were subcutaneously injected with saline solution or 4 mg / kg capsaicin in the neck and back, establishing a solvent control group and an experimental group. Blood was collected 40 minutes later.

[0082] Mice were treated with Hexamethonium bromide (HEX) and injected intraperitoneally with 10 mg / kg HEX. Thirty minutes later, physiological saline or 4 mg / kg capsaicin was injected subcutaneously into the neck and back of the mice. A solvent control group and an experimental group were established. Blood was collected 40 minutes later.

[0083] Blood samples were left at room temperature for 2 hours, then centrifuged at 2000 g for 20 minutes to collect serum. The serum was stored at -80°C before use. The concentration of catecholamines in the serum was detected using the 3-CAT ELISA kit according to the manufacturer's instructions.

[0084] The results are as follows Figure 8 As shown, capsaicin injection increased the expression levels of dopamine, norepinephrine, and epinephrine in the serum of wild-type mice. trpv1 In knockout mice, capsaicin injection did not promote the expression of three catecholamines. HEX treatment can block the binding of acetylcholine to nicotinic acetylcholine receptors secreted by vagal nerve terminals, thereby blocking vagal signal transmission (Ilcol, YO, Cansev, M., Yilmaz, MS, Hamurtekin, E., Ulus, IHPeripheral administration of CDP-choline and its cholinergic metabolites increases serum insulin: muscarinic and nicotinic acetylcholine receptors are both involved in their actions. Neuroscience Letters. 431(1):71-76 (2008)). In mice with subdiaphragmatic vagal nerve transection or HEX-pretreated mice, capsaicin injection did not promote the expression of dopamine and norepinephrine. Therefore, stimulation of TRPV1+ somatic sensory nerves increases catecholamine production by activating the vagus nerve-adrenal axis.

[0085] Example 9: Stimulation of TRPV1+ sensory nerves can activate the hypothalamic-pituitary-adrenal reflex arc and promote the secretion of cortisol hormones. In wild type and trpv1 After subcutaneous injection of solvent or 4 mg / kg capsaicin into the neck and back of knockout mice, blood samples were collected at different time points. The blood samples were left at room temperature for 2 hours, then centrifuged at 2000 g for 20 minutes to collect serum, which was stored at -80°C before use. Serum corticosterone concentration was detected using the Parameter Corticosterone Assay kit according to the manufacturer's instructions.

[0086] The results are as follows Figure 9 As shown, 15 minutes after injecting capsaicin, the concentration of corticosterone in the serum of wild-type mice increased sevenfold, and then gradually decreased, reaching the background level after 3 hours. trpv1 In knockout mice, corticosterone expression did not increase in mice injected with capsaicin.

[0087] The present invention has been described in detail above. Those skilled in the art will recognize that the invention can be practiced in a wide range of ways with equivalent parameters, concentrations, and conditions without departing from its spirit and scope, and without requiring unnecessary experiments. While specific embodiments have been provided, it should be understood that further modifications can be made to the invention. In summary, according to the principles of the invention, this application is intended to include any changes, uses, or improvements to the invention, including changes made using conventional techniques known in the art that depart from the scope disclosed herein.

Claims

1. Use of substances that stimulate TRPV1+ sensory nerves in the preparation of products having any of the following functions: A1) Prevention, treatment, or adjunctive treatment of inflammatory diseases; A2) Inhibit the expression of pro-inflammatory cytokines or inhibit the pro-inflammatory cytokine cascade; A3) Promotes the expression or production of anti-inflammatory cytokines; A4) Prevention, treatment, or adjunctive treatment of autonomic nervous system dysfunction; A5) Promotes catecholamine secretion; A6) Prevention, treatment, or adjuvant treatment of adrenal insufficiency; A7) Promotes the secretion of cortisol hormones by the adrenal glands.

2. The use according to claim 1, characterized in that, The substance that stimulates TRPV1+ sensory nerves is a substance that activates TRPV1 ion channels through chemical stimulation, temperature stimulation, mechanical stimulation, electrical stimulation, ultrasound stimulation, or infrared stimulation.

3. The use according to claim 1 or 2, characterized in that, The substance that stimulates TRPV1+ sensory nerves is a TRPV1 ion channel agonist.

4. The use according to any one of claims 1-3, characterized in that, The TRPV1 ion channel agonist is any one of the following: B1) Reagents or drugs that activate TRPV1 ion channels; B2) Reagents or drugs that activate TRPV1+ sensory neurons; B3) Agents or drugs that promote the replication, transcription, translation, post-transcriptional modification and / or post-translational modification of the TRPV1 gene; B4) Reagents or drugs that increase or upregulate the content, activity, and / or function of TRPV1 protein.

5. The use according to claim 3 or 4, characterized in that, The TRPV1 ion channel agonist is capsaicin or a capsaicin-like substance.

6. The use of TRPV1 ion channels as a target in any of the following: C1) Use in the preparation of products for the prevention, treatment or adjunctive treatment of inflammatory diseases; C2) Use in the preparation of products that inhibit the expression of pro-inflammatory cytokines or inhibit the cascade reaction of pro-inflammatory cytokines; C3) Use in the preparation of products that promote the expression or production of anti-inflammatory cytokines; C4) Use in the preparation of products for the prevention, treatment or adjunctive treatment of autonomic nervous system dysfunction; C5) Use in the preparation of products that promote catecholamine secretion; C6) Use in the preparation of products for the prevention, treatment or adjunctive treatment of adrenal insufficiency; C7) Use in the preparation of products that promote the secretion of cortisol hormones by the adrenal glands; C8) Use in the development, design, or screening of drug candidates and / or treatments for the treatment and / or prevention of inflammatory diseases caused by pro-inflammatory cytokines or pro-inflammatory cytokine cascades; C9) Use in the development, design or screening of candidate drugs and / or treatments for the treatment and / or prevention of autonomic dysfunction disorders and diseases caused by insufficient catecholamine secretion; C10) Use in the development, design, or screening of candidate drugs and / or treatments for the treatment and / or prevention of diseases of adrenal insufficiency.

7. A pharmaceutical composition, characterized in that, The pharmaceutical composition comprises the TRPV1 ion channel agonist and / or TRPA1 ion channel agonist as described in claim 4 or 5, and one or more pharmaceutically acceptable carriers.

8. The use according to any one of claims 1-6, characterized in that, The pro-inflammatory cytokines are TNFα, IL6, IL1a, IL1β, IL1f9, IL1r1, IL1rn, IL2, IL12b, IL13, IL15, IL16, IL20rb, IL27, TNFSF15, TNFSF18, TNFRSF12a, TNFRSF19, CD40, CSF2, CSF3, LIF, TIMP1, TSLP, FAS, OSMR, CCL1, CCL2, CCL3, CCL4, CCL5, CCL7, CCL11, CCL12, CCL17, and C. CL19, CCL20, CCL22, CCL27, CCR6, CXCL1, CXCL5, CXCL10, CXCL11, CX3CL1, XCL1, IFNa1, IFNa2, IFNa4, IFNa5, IFNa9, IFNab, IFNb1, IFNg, IFIH1, IFIT1, IFIT1bl1, IFIT2, IFIT3, IFIT3b, IFI27l2b, IFI47, IFI204, IFI205, IFI207, IFI211, IRF4 and / or IRF6.

9. The use according to any one of claims 1-6, characterized in that, The anti-inflammatory cytokine is IL10.

10. A method for screening candidate drugs for treating and / or preventing inflammatory diseases caused by pro-inflammatory cytokines or pro-inflammatory cytokine cascades, characterized in that, The method includes: screening drugs to be screened with TRPV1 ion channels as targets, and selecting drugs that can activate TRPV1 ion channels or upregulate TRPV1 gene and / or TRPV1 protein levels as candidate drugs.

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