Pharmaceutical composition, organ preserving fluid and application thereof

Abstract

The invention relates to a pharmaceutical composition, an organ preserving fluid and an application of the organ preserving fluid. The pharmaceutical composition comprises telaprevir and borneol. The mass ratio of the telaprevir to the borneol is 1: (0.0001-1000). The pharmaceutical composition disclosed by the invention is good in cell protection effect, telaprevir and borneol show a synergistic effect, excellent curative effects are favorably obtained, the dosage of the medicine is reduced, and the clinical medication safety is favorably improved.

Description

Technical Field

[0001] This invention relates to a pharmaceutical composition for protecting cells, an organ preservation solution, and their applications. The composition can be used to protect individuals (especially at-risk individuals), organs, tissues, or cells from damage and belongs to the field of biomedicine. Background Technology

[0002] During the decline process or pathological conditions, especially those that may lead to cell death, organs, tissues or cells may undergo various pathological processes such as oxidative stress, calcium overload, energy metabolism disorders, and inflammatory responses. These processes can lead to apoptosis, necrosis, and other forms of death in organs, tissues or cells, ultimately resulting in cell death.

[0003] Cellular necrosis includes various pathways such as necroptosis, ferroptosis, and pyroptosis. Studies have shown that RIPK1 / RIPK3 / MLKL-dependent necroptosis exists in various injury-related diseases, such as ischemic stroke, myocardial infarction, hepatic and renal ischemia / reperfusion injury, autoimmune diseases, and neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis (ALS), infantile spinal muscular atrophy, Huntington's disease, and Parkinson's plus syndrome. Inhibiting RIPK1 / RIPK3-dependent necroptosis, ferroptosis, or pyroptosis, such as the RIPK1 inhibitor necrostatin-1 (Nec-1), can reduce the degree of tissue, organ, and cell damage and reduce cell death. In cases of cerebral ischemia / reperfusion injury, the RIPK1 inhibitor necrostatin-1 can reduce cerebral ischemia injury in mice and improve neurological function.

[0004] Oxidative stress (OS) is a state of imbalance between oxidation and antioxidation in the body, with a predominance of oxidation leading to inflammatory infiltration of neutrophils, increased protease secretion, and the production of large amounts of oxidative intermediates. Oxidative stress is a negative effect produced by free radicals in the body and is considered a significant factor contributing to aging and disease. In various injury-related diseases, including ischemic stroke, myocardial infarction, liver and kidney ischemia / reperfusion injury, autoimmune diseases, and neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis (ALS), infantile spinal muscular atrophy, Huntington's disease, and Parkinson's plus syndrome, oxidative stress levels are elevated. Inhibiting oxidative stress can reduce the degree of tissue, organ, and cell damage and decrease cell death.

[0005] If, under multiple pathological conditions, such as ischemia / reperfusion injury of the heart and brain, apoptosis, necroptosis-like apoptosis, ferroptosis, or pyroptosis pathways are simultaneously inhibited, cell death can be synergistically suppressed, greatly reducing the degree of tissue, organ, and cell damage, decreasing cell death, and simultaneously reducing the dosage and adverse reactions of individual drugs. However, existing compounds that inhibit these pathways can only serve as tool drugs and cannot meet clinical needs.

[0006] Therefore, it is essential to find novel drug compositions (cell protectants) with greater clinical application potential for cell protection.

[0007] Telaprevir is an inhibitor of hepatitis C virus (HCV) NS3 / 4A serine protease, possessing antiviral activity by inhibiting HCV replication. Previous research by our team has found that telaprevir can be used to prepare drugs for the treatment or prevention of ischemia / reperfusion injury (see CN202110943155.9). Borneol, also known as camphor, 2-borneol, borneol, and borneol tablets, has anti-inflammatory and sedative effects. However, whether the combined use of these two drugs has a synergistic effect is unclear, and no relevant reports have been found. Summary of the Invention

[0008] In this invention, the term "cell protection" refers to the effect of any reagent or compound (whether natural or non-natural) in protecting and / or preventing and / or treating cells against the consequences of pathological conditions (particularly those that may lead to cell death) or degenerative processes at the cellular level.

[0009] In this invention, unless otherwise stated, "pathological condition" refers to, for example, symptoms or disease or trauma or exposure to various factors (especially factors that trigger cell death processes), and also includes events such as bleeding, accidental occlusion (infarction) and / or medical procedures (especially surgical procedures, such as organ transplantation).

[0010] In this invention, "protection" refers to preventing the occurrence of pathological conditions or degenerative processes that may lead to cell death at the cellular level (especially for at-risk individuals), or inhibiting, reducing, or treating such consequences.

[0011] In this invention, "treatment" refers to preventive (advantageously for individuals at risk) and / or alleviating and / or curative treatment. It includes: a) inhibiting and / or eliminating the occurrence and / or development of a degenerative process or pathological condition; b) or mitigating the severity of such a degenerative process or pathological condition, for example, reducing the frequency or severity of symptoms associated with the degenerative process or pathological condition, improving the quality of life of individuals suffering from such a degenerative process or pathological condition, reducing the dosage of other medications required to treat the degenerative process or pathological condition, enhancing the effectiveness of another treatment for the degenerative process or pathological condition, or prolonging the lifespan of individuals suffering from such a degenerative process or pathological condition.

[0012] "Prevention" or "avoidance" refers to reducing the likelihood of development or preventing or delaying the occurrence of a decline process or pathological condition in an individual who has not yet developed such a condition but is at risk of developing one.

[0013] "At risk" means that an individual has one or more risk factors for a degenerative process or pathology, which are measurable parameters that can be associated with the development of the degenerative process or pathology and are known to those skilled in the art. Individuals exhibiting one or more of these risk factors are more likely to develop a degenerative process or pathology than individuals who do not exhibit these risk factors. For example, an individual planning to undergo surgery may be considered an at-risk individual. As another example, an individual with the following risk factors may be considered an individual with risk factors for stroke and cerebral ischemia: hypertension, carotid artery stenosis, transient ischemic attack, coronary artery disease, history of myocardial infarction, lack of physical activity, atrial fibrillation, left ventricular dysfunction or mitral stenosis, heart failure, hyperlipidemia, smoking, and diabetes.

[0014] The terms "organ," "tissue," or "cell" refer to one or more cells, a part of an organ, an entire organ, tissue, or group of tissues (limbs, etc.) of human or animal origin. This invention may be applicable to all organs, tissues, or cells. Examples include: solid organs such as the heart, liver, brain, lungs, kidneys, pancreas, intestines, and eyes; cells such as cells or stem cells of the aforementioned organs; and tissues such as skin, cornea, and vascularized composite tissue (VCA). Preferably, this invention targets solid organs; even more preferably, the brain, heart, intestines, lungs, liver, and kidneys are the targets of this invention.

[0015] In this invention, unless otherwise stated, telaprevir refers to a telaprevir compound (drug) or a pharmaceutically acceptable salt or ester of the same or an isomer thereof or a semi-synthetic derivative thereof or a salt of the same or an ester thereof or an ester salt of the same or an ester salt thereof or a deuterated compound thereof (deuterated telaprevir) or an isotopically labeled compound thereof.

[0016] Bornol, also known as camphor, 2-borneol, borneol, borneol, and borneol, includes dextroborneol [(+)-borneol, also known as (+)-2-borneol, dextroborneol], levonborneol [(-)-borneol, also known as (-)-2-borneol, levonborneol] and racemic borneol [(±)-borneol, also known as (±)-2-borneol].

[0017] Optionally, camphor includes one or more of dexborneol (or natural borneol), levborneol, and racemic camphor.

[0018] Optionally, the pharmaceutically acceptable salt is a pharmaceutically commonly used salt, and further, the salt is selected from one or more of the following: acetate, hydrochloric acid, hydrobromic acid, nitric acid, sulfuric acid, phosphoric acid, benzoate, fumarate, maleate, succinic acid, tartaric acid, citrate, oxalic acid, glyoxylic acid, aspartic acid, tartrate, 2,5-dihydroxybenzoate, methanesulfonate, ethanesulfonate, benzenesulfonate, lecithin sulfonate, hydroquinone sulfonate, and p-toluenesulfonate, or salts formed from carboxylic acids (e.g., formic acid, acetic acid, or propionic acid).

[0019] Generally, the structural formula of telaprevir is shown in Formula I, and its molecular formula is C10. 36 H 53 N7O6:

[0020]

[0021] The structural formula of dextromethorphan is shown in Formula II, and its molecular formula is C. 10 H 18 O.

[0022]

[0023] The structural formula of levonorhamnetin is shown in Formula III, and its molecular formula is C. 10 H 18 O.

[0024]

[0025] The structural formula of racemic camphor is shown in Formula IV, and its molecular formula is C10. 10 H 18 O.

[0026]

[0027] To address the shortcomings of existing technologies, one objective of this invention is to provide a pharmaceutical composition capable of protecting, preventing, and / or treating cells against processes that lead to cell death, such as drugs against pathological apoptosis and / or necrosis and / or necroptosis and / or ferroptosis and / or disulfide death and / or autophagy, or pharmaceutical compositions that resist surgical procedures that may lead to cell death. Another objective of this invention is to provide the application of the pharmaceutical composition in the preparation of drugs that protect, prevent, and / or treat cells against processes that lead to cell death.

[0028] More specifically, in view of the shortcomings of the prior art, the present invention aims to provide a pharmaceutical composition for protecting cells with superior efficacy; a second objective of the present invention is to provide the application of the above-mentioned pharmaceutical composition in the preparation of cell protection drugs; a third objective of the present invention is to provide the application of the above-mentioned pharmaceutical composition in the preparation of organ preservation solutions; and a fourth objective of the present invention is to provide an organ preservation solution.

[0029] To solve the above-mentioned technical problems, the technical solution of the present invention is as follows:

[0030] A pharmaceutical composition comprising terabhivir and camphene.

[0031] Further, the mass ratio of terabhivir to camphene is 1:0.0001-10000, preferably 1:0.001-1000, more preferably 1:0.01-100, even more preferably 1:0.025-50, more preferably 1:0.03-45, even more preferably 1:0.05-25, even more preferably 1:0.007-20, even more preferably 1:0.1-10, even more preferably 1:0.1-5, even more preferably 1:0.15-1, and even more preferably 1:0.2-0.8.

[0032] Further, the mass ratio of terabhivir to camphor is 5-40:1-10, preferably 10-35:2-8, more preferably 15-30:3-6, even more preferably 18-25:4-5, and even more preferably 19:4-5, 20:4-5, 21:4-5, 22:4-5, 23:4-5 or 24:4-5.

[0033] Further, the mass ratio of terabhivir to camphor is 3-30:0.25-10, preferably 5-30:0.5-8, more preferably 7.5-30:0.75-6, even more preferably 10-30:0.8-2, and even more preferably 12-30:0.9-1.5.

[0034] Optionally, in the pharmaceutical composition, the mass ratio of telaprevir to camphor is 20:7, 19:7, 18:7, 17:7, 16:7, 15:7, 14:7, 12:7, 11:7, 10:7, 9:7, 8:7, 7:7, 6:7, 5:7, 20:6, 19:6, 18:6, 17:6, 16:6, 15:6, 14: 6, 12:6, 11:6, 10:6, 9:6, 8:6, 7:6, 6:6, 5:6, 20:5, 19:5, 18:5, 17:5, 16:5, 15:5, 14:5, 12:5, 11:5, 10:5, 9:5, 8:5, 7:5, 6:5, 5:5, 20:4, 19:4, 18:4, 17:4, 16 :4, 15:4, 14:4, 12:4, 11:4, 10:4, 9:4, 8:4, 7:4, 6:4, 5:4, 20:3, 19:3, 18:3, 17:3, 16:3, 15:3, 14:3, 12:3, 11:3, 10:3, 9:3, 8:3, 7:3, 6:3, 5:3, 20:2, 19:2, 1 8:2, 17:2, 16:2, 15:2, 14:2, 12:2, 11:2, 10:2, 9:2, 8:2, 7:2, 6:2, 5:2, 20:1, 19:1, 18:1, 17:1, 16:1, 15:1, 14:1, 12:1, 11:1, 10:1, 9:1, 8:1, 7:1, 6:1 or 5:1.

[0035] Optionally, the mass ratio of telaprevir to camphene in the pharmaceutical composition is 30:0.1, 30:0.15, 30:0.2, 30:0.25, 30:0.3, 30:0.35, 30:0.4, 30:0.45, 30:0.5, 30:0.55, 30:0.6, 30:0.65, 30:0.7, 30:0.75, 30:0.8, 30:0.85, 30:0.9, 30:0.95, 30:1, 30:1.05, 30:1.1, 30:1.15, 30:1.2, 30:1.25, 30:1.3, 30:1.35, 30:1.4, 30:1.45, or 30:1.5.

[0036] Furthermore, the molar ratio of terabhivir to camphene is 5-75:3-65, preferably 6-70:4-60, and more preferably 7-68:5-58.

[0037] Furthermore, the pharmaceutical composition further includes a pharmaceutically acceptable carrier. Furthermore, the active ingredients of the pharmaceutical composition may comprise or be telaprevir and camphene.

[0038] Furthermore, the pharmaceutical composition can be prepared into any pharmaceutically acceptable dosage form according to known techniques. The pharmaceutical composition of the present invention can be formulated into the following forms: suspension or ready-to-use injection solution or temporary injection solution, gel, oil, tablet, suppository, powder, capsule, granule, suspension, emulsion, polymer, nanoparticle, microsphere, rectal capsule, enema, paste, ointment, cream, plaster, decoction, implant, spray, aerosol, etc., optionally with controlled release and / or sustained release via dosage form or device. Preferred dosage forms include injection, capsule, tablet, granule, powder, spray, liposome, oral liquid, and pellet.

[0039] The pharmaceutical combination of terabhivir and camphor can be used (advantageously in at-risk individuals) for the prevention and / or protection and / or treatment of humans and / or animals (particularly mammals, preferably humans).

[0040] Based on the same inventive concept, the present invention also provides the use of the pharmaceutical composition described above in the preparation of cell protection drugs.

[0041] Further, the cell-protective drug is a drug used to prevent, protect, and / or treat cell damage caused by pathological conditions and / or degeneration processes; the pathological conditions and / or degeneration processes include situations that may lead to cell death, preferably, the pathological conditions and / or degeneration processes include pathological apoptosis and / or pathological necrosis and / or necrotizing apoptosis and / or pyroptosis and / or ferroptosis and / or disulfide-dependent cell death and / or autophagy (anti-apoptotic drugs and / or anti-necrotizing drugs and / or anti-pyroptosis drugs and / or anti-ferroptosis drugs and / or anti-dependent cell death drugs and / or disulfide-dependent cell death drugs and / or anti-autophagy drugs) and / or diseases or conditions; preferably, the diseases or conditions include, but are not limited to: nervous system diseases, cardiovascular system diseases The following conditions are contraindicated: hemorrhage and thrombotic diseases, diffuse connective tissue diseases, organ-specific or systemic inflammation or autoimmune diseases, autoimmune diseases, bone diseases, joint and cartilage diseases, ischemic diseases or attacks of limbs, ophthalmic diseases, skin diseases, kidney diseases, blood and vascular diseases, lung diseases, gastrointestinal diseases, liver diseases, metabolic diseases, muscle diseases, pancreatic diseases, severe poisoning caused by chemicals, infectious agents, toxins or drugs, age-related diseases, dental diseases, auditory conduction pathway diseases, mitochondrial-related diseases, and / or trauma and / or exposure to factors of biological and / or chemical and / or physical and / or medical and / or surgical procedures, such as accidental infarction and hemorrhage, and / or medical and / or surgical procedures, such as cell, tissue or organ transplantation. The cell-protective drugs mentioned refer to drugs that have the effect of preventing, inhibiting or treating damage, degeneration or dysfunction of tissues, organs and cells caused by hypoxia / reoxygenation.

[0042] Alternatively, the cell-protective drug refers to a drug that has the effect of preventing, inhibiting or treating damage, degeneration or dysfunction of tissues, organs and cells caused by necroptosis.

[0043] Furthermore, the neurological diseases mentioned include stroke, transient ischemic attack, focal ischemia, intracranial hemorrhage, prenatal hypoxia, adult or childhood hypoxia, neurodegenerative diseases, muscle diseases, trigeminal neuralgia, glossopharyngeal neuralgia, Bell's palsy, progressive bulbar palsy, primary lateral sclerosis (PLS), pseudobulbar palsy, invertebral disc syndrome, cervical spondylosis, plexus disorder, thoracic outlet destruction syndrome, porphyria, peripheral neuropathy, multiple system atrophy, corticobasal degeneration, progressive supranuclear palsy, Lewy body dementia, demyelinating diseases, frontotemporal dementia, Gullman-Barré syndrome, multiple sclerosis, Kreutzfeldt-Jacob disease, progressive peroneal muscular atrophy, prion disease, and lethal... Familial insomnia, G.S.-S. syndrome, bovine spongiform encephalopathy, epilepsy, hereditary ataxia, Friedreich ataxia, spinocerebellar ataxia, hereditary spastic paraplegia, dystonia, multiple system atrophy, lysosomal storage disease, Niemann-Pick disease, Gaucher disease, AIDS dementia syndrome, neurological damage caused by exposure to toxic compounds consisting of industrial solvents, heavy metals, drugs, and chemotherapeutic agents, and neurological damage caused by mechanical, physical, or chemical trauma; preferably, stroke includes one or more of ischemic and hemorrhagic strokes; preferably, the neurodegenerative disease includes Alzheimer's disease. Muscle diseases include one or more of the following: Parkinson's disease, amyotrophic lateral sclerosis (ALS), infantile spinal muscular atrophy, Huntington's disease, and Parkinson's plus syndrome; preferably, the muscle diseases include one or more of the following: muscular atrophy, Duchenne's muscular dystrophy, ankylosing spondylitis, myopathy and myasthenia gravis, myasthenia gravis, progressive muscular atrophy, spinal muscular atrophy, and hereditary muscular atrophy;

[0044] Preferably, the cardiovascular diseases include one or more of the following: myocardial ischemia and / or vascular ischemia, ischemic heart disease, angina pectoris, unstable angina pectoris, refractory angina pectoris, myocardial infarction, myocardial ischemia / reperfusion injury, hypoxia, low oxygen, chronic or acute heart failure, systolic heart failure and diastolic heart failure, left ventricular dysfunction, post-myocardial infarction left ventricular dysfunction, hypertensive heart disease, rheumatic heart disease, cardiomyopathy, myocardial hypertrophy, hypertrophic cardiomyopathy, myocarditis, valvular heart disease, arrhythmia, paroxysmal tachycardia, atrial fibrillation, ventricular fibrillation, arteriosclerosis, atherosclerosis, peripheral vascular disease, aneurysm, peripheral vascular occlusive disease (preferably cerebral occlusion, pulmonary occlusion or intestinal occlusion), chronic venous insufficiency or varicose veins, hypertension, systemic hypertension, pulmonary hypertension, portal hypertension, and cardiovascular toxic side effects caused by drug (preferably anticancer drugs) treatment;

[0045] Preferably, the stroke includes ischemic stroke;

[0046] Preferably, the diffuse connective tissue disease includes one or more of the following: rheumatoid arthritis, juvenile idiopathic arthritis, lupus erythematosus, systemic lupus erythematosus, scleroderma, idiopathic inflammatory myopathy, polymyositis, dermatomyositis, vasculitis, necrotizing vasculitis, polyarteritis nodosa, granulomatous vasculitis, giant cell arteritis, Sjögren's syndrome, systemic scleroderma, allergic cutaneous vasculitis, and Behcet's disease;

[0047] Preferably, the organ-specific inflammation, systemic inflammation, or autoimmune disease includes one or more of chronic inflammatory bowel disease, bronchial asthma, chronic obstructive pulmonary disease, eosinophilic sinusitis, and systemic lupus erythematosus.

[0048] Preferably, the ophthalmic diseases or conditions include one or more of the following: diabetic retinopathy, glaucoma, retinal degeneration, retinitis pigmentosa, corneal reticular dystrophy, optic neuropathy and optic neuritis, optic drusen, ptosis, chronic progressive extraocular muscle paralysis, macular degeneration, retinal hole or retinal tear, retinal ischemia, retinal ischemia / reperfusion injury, retinal detachment, trauma-related acute retinopathy, inflammatory degeneration, postoperative complications, drug-induced retinopathy or cataract, wet or dry AMD-related photoreceptor degeneration;

[0049] Preferably, the skin disease includes one or more of the following: dermatitis, psoriasis, scarring, aging or altered healing process, eczema, and collagen diseases;

[0050] Preferably, the trauma and / or exposure to factors of biological and / or chemical and / or physical and / or medical and / or surgical origins include severe poisoning caused by infectious agents, toxins, chemicals, or drugs. More preferably, the symptoms of severe poisoning include one or more of sepsis, septic shock and its consequences, or iatrogenic diseases.

[0051] Furthermore, the cell-protective drug is one or more drugs used to prevent and / or protect and / or treat cell death in transplanted organs and / or organ donors and / or organ recipients, prevent acute transplant rejection of organs and / or increase long-term survival rates, limit primary organ dysfunction and / or limit the delay in the recovery of transplanted organ function and / or improve the functional recovery of transplanted organs; primarily preventing or treating cell death in transplanted organs.

[0052] Advantageously, pharmaceutical compositions containing telaprevir and camphene can be used before, during, or after transplantation for living or clinically dead organ donors, tissue donors, cell donors, organ recipients, tissue recipients, or cell recipients; and / or more specifically, the organs, tissues, or cells can be not only in situ organs, tissues, or cells (e.g., in medicine, during surgery, or in pathological processes) but also ex vivo organs, tissues, or cells (e.g., in certain procedures requiring temporary removal of organs, tissues, or cells from the body, particularly those that modify or purify them, or during the transport and preservation of organs, tissues, or cells, during transplantation, or during their reperfusion after reimplantation).

[0053] Therefore, this prophylaxis and protection can be applied in a general manner to individuals, donors or recipients, or organs, tissues or cells, in situ or ex vivo, for example during certain surgeries, or during their transport or preservation for reimplantation.

[0054] Ischemia primarily results from a reduced or interrupted blood supply to an organ, leading to decreased blood, oxygen, and energy supply to organs, tissues, and cells. This causes damage and death of tissues, organs, and cells, and is associated with, but not limited to, atherosclerotic plaques, thrombosis, arterial compression (e.g., through limb compression, tourniquet use, tumors, hematomas, or fluid leakage), artificial cessation of blood circulation (if necessary during surgery), hemorrhage, or insufficient perfusion. Ischemia or insufficient perfusion can affect or damage the function of all organs, particularly the brain, heart, liver, lungs, kidneys, intestines, or limbs. When blood circulation to an organ is restored after a period of ischemia, reperfusion can also cause damage to the organ, tissues, and cells, a condition known as ischemia / reperfusion injury. This can limit the recovery of function and may even jeopardize an individual's survival. Ischemia / reperfusion injury involves multiple types of cell death.

[0055] Unexpectedly, the inventors discovered that a pharmaceutical composition containing terabitvir and camphor can reduce damage to human or animal cells from different types of organs subjected to organ ischemia / reperfusion and increase their survival rate.

[0056] Preferably, the cell-protective drug is a drug for preventing and / or protecting and / or treating pathological conditions or degenerative processes associated with ischemia / reperfusion symptoms (especially those that lead to cell death). More preferably, the pathological conditions or degenerative processes associated with ischemia / reperfusion symptoms include one or more of actual cold ischemia, warm ischemia, actual reperfusion, and ischemia / reperfusion phenomena.

[0057] Preferably, the cell-protective drug is a drug used to prevent and / or protect and / or treat organs, tissues or cells against ischemia / reperfusion injury. Optionally, the ischemia / reperfusion injury is caused by cold or warm ischemia and / or reperfusion and / or ischemia / reperfusion. Further, the ischemia / reperfusion injury includes one or more of cerebral ischemia / reperfusion injury, myocardial ischemia / reperfusion injury, hepatic ischemia / reperfusion injury, renal ischemia / reperfusion injury, pulmonary ischemia / reperfusion injury, intestinal ischemia / reperfusion injury and limb ischemia / reperfusion injury.

[0058] In a preferred form, the pharmaceutical composition containing telaprevir and camphenol can be used as a medicine for the prevention and / or protection and / or treatment of: neurological sequelae due to stroke or trauma, heart failure due to infarction, tissue damage affecting the heart, liver, intestine, lung or kidney after transplantation to a graft or surgery, or damage caused by surgical procedures.

[0059] A more preferred form is a pharmaceutical composition containing telaprevir and camphene, which can be used as a medicine for the prevention and / or protection and / or treatment of ischemic stroke caused by actual hot or cold ischemia and / or actual reperfusion and / or ischemia / reperfusion.

[0060] A more preferred form is a pharmaceutical composition containing telaprevir and camphene, which can be used as a medicine for the prevention and / or protection and / or treatment of myocardial infarction caused by actual hot or cold ischemia and / or actual reperfusion and / or ischemia / reperfusion.

[0061] According to the present invention, the pharmaceutical composition containing terabhivir and camphene is used in a physiologically effective amount.

[0062] As a medicine, the pharmaceutical composition containing telaprevir and camphor can be formulated for use in the digestive tract or external gastrointestinal tract.

[0063] Furthermore, the cell-protective drug is a drug used to prevent and / or protect and / or treat one or more of the following: heart failure caused by infarction, neurological sequelae caused by stroke or trauma, tissue damage affecting liver, intestine, heart, lung or kidney transplantation to grafts or surgery, or consequences of surgical procedures.

[0064] Alternatively, the cell-protective drug is a drug for preventing and / or protecting and / or treating one or more of nerve cells (a drug for protecting the brain), heart cells (a drug for protecting the heart), liver cells (a drug for protecting the liver), kidney cells (a drug for protecting the kidneys), intestinal cells, or lung cells. Preferably, the cell-protective drug is a drug for protecting nerve cells, vascular endothelial cells, brain endothelial cells, and / or cardiomyocytes.

[0065] Optionally, the factors that trigger the cell death process may be of biological and / or chemical and / or physical origin.

[0066] The application of the pharmaceutical composition described above in the preparation of organ preservation solutions.

[0067] Optionally, biological sources include, for example, asphyxiation, ischemia / reperfusion, hypoxia or oxygen deprivation, nutrient deprivation, poisoning from in-situ generated free radicals or reactive oxygen species, growth factor deficiency, and massive release of cytotoxins or cytokines. They may also originate from events such as hemorrhage, accidental occlusion (infarction), and certain medical procedures (e.g., cuff inflation, ventilator use, sutures), as well as biological or chemical agents used as therapeutic agents in medical treatment (e.g., immunosuppressants, cell inhibitors or cytotoxic agents, anti-inflammatory drugs).

[0068] Optionally, chemical sources include, for example, poisoning from toxins, waste, pH changes, free radicals, reactive oxygen species, and environmental toxins.

[0069] Alternatively, physical sources include, for example, impact, cuts, exposure to radiation (X-ray radiation, gamma radiation, UV radiation, etc.), hyperthermia, hypothermia, or the presence of foreign objects or crystals in the organism.

[0070] Alternatively, surgical procedures that may lead to cell death processes may include, for example, procedures that require a brief interruption of blood circulation resulting in systemic or local ischemia or hypoperfusion (such as the use of tourniquets, hemostatic forceps, etc.), occurring, for example, during surgery, particularly during angioplasty of organs or the heart or main or peripheral blood vessels, or thoracic surgery, cardiac surgery, or vascular surgery that sometimes requires bypassing the cardiopulmonary system or stopping the heart, and any surgery that requires voluntary obstruction of an organ or part of an artery or reduction of blood flow through an organ.

[0071] Optionally, very important diseases or symptoms that may lead to cell death processes include (but are not limited to) the following diseases or symptoms (which are often accompanied by apoptosis and / or necrosis and / or necroptosis and / or ferroptosis and / or pyroptosis and / or autophagy):

[0072] Neurological disorders include stroke, transient ischemic attack, focal ischemia, intracranial hemorrhage, prenatal hypoxia, hypoxic-ischemic encephalopathy in adults or children, neurodegenerative diseases, muscle diseases, trigeminal neuralgia, glossopharyngeal neuralgia, Bell's palsy, progressive bulbar palsy, primary lateral sclerosis (PLS), pseudobulbar palsy, invertebral disc syndrome, cervical spondylosis, plexus disorder, thoracic outlet destruction syndrome, porphyria, peripheral neuropathy, multiple system atrophy, corticobasal degeneration, progressive supranuclear palsy, Lewy body dementia, demyelinating diseases, frontotemporal dementia, Gullman-Barré syndrome, multiple sclerosis, Kreutzfeldt-Jacob disease, progressive peroneal muscular atrophy, prion disease, and fatal familial insomnia. The following are included in the list of neurodegenerative diseases: Göttingen-Schwarz syndrome, bovine spongiform encephalopathy, epilepsy, hereditary ataxia, Friedreich-type ataxia, spinocerebellar ataxia, hereditary spastic paraplegia, dystonia, multiple system atrophy, lysosomal storage diseases, Niemann-Pick disease, Gaucher disease, AIDS dementia syndrome, neurological damage caused by exposure to toxic compounds in the group consisting of industrial solvents, heavy metals, drugs and chemotherapeutic agents, and neurological damage caused by mechanical, physical or chemical trauma; preferably, stroke includes one or more of ischemic stroke and hemorrhagic stroke; preferably, the neurodegenerative disease includes Alzheimer's disease. Muscle diseases include one or more of the following: Parkinson's disease, amyotrophic lateral sclerosis (ALS), infantile spinal muscular atrophy, Huntington's disease, and Parkinson's plus syndrome; preferably, the muscle diseases include one or more of the following: muscular atrophy, Duchenne's muscular dystrophy, ankylosing spondylitis, myopathy and myasthenia gravis, myasthenia gravis, progressive muscular atrophy, spinal muscular atrophy, and hereditary muscular atrophy;

[0073] Pain, such as neuropathic pain, inflammatory pain, and diabetic pain;

[0074] Cardiovascular diseases, such as myocardial ischemia and / or vascular ischemia, ischemic heart disease, angina pectoris, unstable angina pectoris, refractory angina pectoris, myocardial infarction, myocardial ischemia / reperfusion injury, hypoxia, hypoxia, chronic or acute heart failure, systolic heart failure and diastolic heart failure, left ventricular dysfunction, post-myocardial infarction left ventricular dysfunction, hypertensive heart disease, rheumatic heart disease, cardiomyopathy, myocardial hypertrophy, hypertrophic cardiomyopathy, myocarditis, valvular heart disease, arrhythmia, paroxysmal tachycardia, atrial fibrillation, ventricular fibrillation, arteriosclerosis, atherosclerosis, peripheral vascular disease, aneurysm, peripheral vascular occlusive disease (especially cerebral occlusion, pulmonary occlusion or intestinal occlusion), chronic venous insufficiency or varicose veins, hypertension, systemic hypertension, pulmonary hypertension, portal hypertension, cardiovascular toxicity due to drug (especially anticancer drugs) treatment;

[0075] Myocardial remodeling includes myocardial remodeling after myocardial ischemia / hypoxia (such as myocardial infarction), myocardial remodeling after cardiac surgery, myocardial remodeling after arterial valve disease, vascular hypertrophy (smooth muscle cell hypertrophy), and vascular remodeling.

[0076] Hemorrhage and thrombotic diseases, thromboembolic diseases and venous thrombosis, vascular permeability disorders, restenosis, acute coronary syndrome, venous embolism occurring after thrombolytic therapy or coronary angioplasty, venous thrombosis, pulmonary embolism caused by venous thrombosis, cerebrovascular syndromes such as embolic stroke, transient ischemic attack, and occlusive coronary thrombosis. Coagulopathy, thrombotic thrombocytopenic purpura, disseminated intravascular coagulation, thromboangiitis obliterans, and thrombotic diseases related to heparin-induced thrombocytopenia. Thrombotic complications related to cardiopulmonary bypass, thrombotic complications related to instruments such as cardiac or other endovascular catheterization, intraaortic balloon catheterization, coronary stenting, or cardiac valves, and conditions requiring assistive devices and similar symptoms.

[0077] Diffuse connective tissue diseases, such as rheumatoid arthritis, juvenile idiopathic arthritis, lupus erythematosus, systemic lupus erythematosus, scleroderma, idiopathic inflammatory myopathy, polymyositis, dermatomyositis, vasculitis, necrotizing vasculitis, polyarteritis nodosa, granulomatous vasculitis, giant cell arteritis, Sjögren's syndrome, systemic scleroderma, allergic cutaneous vasculitis, Behcet's disease, etc.

[0078] Organ-specific inflammation or systemic inflammation or autoimmune diseases, such as chronic inflammatory bowel disease, bronchial asthma, chronic obstructive pulmonary disease, and eosinophilic sinusitis or systemic lupus erythematosus.

[0079] Autoimmune diseases or symptoms, including but not limited to Hashimoto's thyroiditis, autoimmune atrophic gastritis, autoimmune orchitis, autoimmune encephalomyelitis, autoimmune thrombocytopenia, autoimmune alopecia, ulcerative colitis, hemolytic anemia, pernicious anemia, sympathetic ophthalmia, Graves' disease, primary biliary cirrhosis, chronic invasive hepatitis, conjunctival tetragonal lesions, and systemic lupus erythematosus.

[0080] Bone diseases, joint diseases, and cartilage diseases, such as osteoporosis, osteomyelitis, ischemic necrosis, arthritis (including osteoarthritis and psoriatic arthritis), spondyloarthritis, ankylosing spondylitis, rickets, progressive ossifying fibrous proliferative disease, and Cushing's syndrome.

[0081] Ischemic limb disease or attack;

[0082] Ophthalmic diseases or conditions such as diabetic retinopathy, glaucoma, retinal degeneration, retinitis pigmentosa, corneal reticular dystrophy, optic neuropathy and optic neuritis, optic drusen, ptosis, chronic progressive extraocular muscle paralysis, macular degeneration, retinal hole or retinal tear, retinal ischemia, retinal ischemia / reperfusion injury, retinal detachment, trauma-related acute retinopathy, inflammatory degeneration, postoperative complications, drug-induced retinopathy or cataracts, wet or dry AMD-related photoreceptor degeneration;

[0083] Skin conditions such as dermatitis, psoriasis, scarring, changes in the aging or healing process, eczema, and collagen diseases;

[0084] Kidney diseases, such as renal fibrosis, acute kidney disease, renal ischemia, renal capillary infarction, acute kidney injury, acute or chronic interstitial nephropathy, glomerulonephritis, diabetic nephritis, renal artery sclerosis, renal insufficiency, acute or chronic renal failure or dialysis side effects, and renal failure after myocardial ischemia / reperfusion.

[0085] Blood disorders, such as anemia, bleeding, angioamyloidosis, sickle cell disease, neutropenia, erythrocyte fragmentation syndrome, pancytopenia, leukopenia, myelodysplastic syndrome, thrombocytopenia, and hemophilia;

[0086] Lung diseases, such as pulmonary hypertension, acute respiratory distress syndrome, respiratory infections, chronic obstructive pulmonary disease (COPD), such as chronic bronchitis and emphysema, asthma, cystic fibrosis; pulmonary cystic disease,

[0087] Gastrointestinal diseases, such as chronic inflammatory bowel disease, ulcers or mesenteric infarction, portal hypertension;

[0088] Liver diseases, such as autoimmune hepatitis, viral hepatitis or hepatitis caused by other infectious agents, liver fibrosis, alcoholic liver disease (ALD), alcoholic hepatitis, fulminant hepatitis, cirrhosis, and liver diseases caused by toxins or drugs.

[0089] Steatosis, such as liver ischemia or drug-induced exogenous poisoning, alcoholic or non-alcoholic steatohepatitis (NASH);

[0090] Metabolic diseases, such as diabetes, diabetic complications, diabetic nephropathy, diabetic retinopathy, diabetic foot, thyroiditis, Hashimoto's thyroiditis, glucose intolerance syndrome, obesity, abeta-lipoproteinemia, hyperlipidemia, hypothalamic-pituitary axis dysfunction, diabetes insipidus, galactosemia, glycogenopathies, gout, Wilson's disease, or Weber-Christian disease;

[0091] Pancreatic diseases, such as chronic pancreatitis or acute pancreatitis;

[0092] Severe poisoning caused by infectious agents, toxins, chemicals or drugs, such as sepsis, septic shock and its consequences or iatrogenic diseases;

[0093] Age-related diseases, such as accelerated aging syndrome;

[0094] Dental diseases, such as those that cause tissue damage, such as periodontitis;

[0095] Diseases of the auditory conduction pathway, such as antibiotic-induced deafness and otosclerosis;

[0096] Mitochondrial-related diseases (mitochondrial pathology), such as congenital muscular dystrophy with structural mitochondrial abnormalities and Friedrich's ataxia.

[0097] And / or trauma and / or exposure to factors of biological and / or chemical and / or physical origin and / or events, such as accidental bleeding and infarction, and / or medical procedures and / or surgical procedures, such as cell, tissue or organ transplantation.

[0098] Advantageously, the above-described pharmaceutical composition is used, for example, for the prevention and / or protection and / or treatment of nerve cells (a drug for protecting brain cells), cardiomyocytes (a drug for protecting the heart), lungs (a drug for protecting the lungs), intestines (a drug for protecting the intestines), liver (a drug for protecting the liver), and kidneys (a drug for protecting the kidneys), preferably for the protection of nerve cells (a drug for protecting the brain), cardiomyocytes (a drug for protecting the heart), and intestines (a drug for protecting the intestines), and very preferably for nerve cells and cardiomyocytes.

[0099] The pharmaceutical compositions of the present invention, comprising telaprevir and camphene, are intended for simultaneous, separate, or time-interval use as medicines, particularly for the prevention and treatment of individuals at risk and / or for individuals suffering from at least one pathological condition or at least one of the aforementioned degenerative processes. It should be understood that, for simultaneous use, the compounds present in the pharmaceutical compositions comprising the compounds may be mixed together or physically separated; for separate or time-interval use, the present compounds must be physically separated.

[0100] The pharmaceutical compositions of the present invention, comprising telaprevir and camphene, are used concurrently, separately, or at time intervals as cell-protective drugs or medicines for the prevention and / or protection and / or treatment of the cellular consequences of the aforementioned pathological conditions or degenerative processes.

[0101] The pharmaceutical compositions of the present invention, comprising telaprevir and camphene, are used concurrently, separately, or at time intervals as cellular protective agents or medicines for the prevention and / or protection and / or treatment of the consequences of the aforementioned pathological conditions or degenerative processes at the cellular level and / or for the treatment of subjects suffering from one of the aforementioned pathological conditions or degenerative processes.

[0102] In a highly preferred form, the present invention relates to pharmaceutical compositions comprising telaprevir, camphene, or a pharmaceutically acceptable salt or ester thereof, or an isomer thereof, or a semi-synthetic derivative thereof, or a salt thereof, or an ester thereof, or a deuterated compound thereof, or an isotopically labeled compound thereof, for use as a cellular protective agent or medicine for preventing and / or protecting and / or treating the consequences of the aforementioned pathological conditions or degenerative processes at the cellular level and / or treating a subject suffering from one of the aforementioned pathological conditions or degenerative processes.

[0103] Optionally, with respect to this aspect of the invention relating to the composition, consideration should also be given to the use of terabhivir and camphor in the pharmaceutical compositions of the invention as cell-protective agents, as well as the implementation methods and definitions relating to administration and dosage.

[0104] The pharmaceutical composition of the present invention, comprising telaprevir and camphene, is for use concurrently, separately, or at time intervals as a medicine for preventing and / or protecting and / or treating cells, tissues, and organs against ischemia / reperfusion injury (which may occur during actual cold or hot ischemia and / or actual reperfusion and / or ischemia / reperfusion).

[0105] In a highly preferred form, the present invention relates to pharmaceutical compositions as medicines for preventing and / or protecting and / or treating cells, tissues, and organs against ischemia / reperfusion injury (which may occur during actual cold or hot ischemia and / or actual reperfusion and / or ischemia / reperfusion).

[0106] In a highly preferred form, the present invention relates to pharmaceutical compositions as remedies for preventing and / or protecting and / or treating cells, tissues, and organs against degeneration and degenerative changes such as neurodegenerative diseases (e.g., Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis (ALS), infantile spinal muscular atrophy, Huntington's disease, Parkinson's plus syndrome, and multiple sclerosis).

[0107] According to one embodiment of the present invention, the amount of terabhivir in the pharmaceutical composition may be from 0.001 mg to 6000 mg / mg camphor, preferably from 1 mg to 600 mg (terabhivir) / mg camphor.

[0108] According to the present invention, telaprevir and camphor in the pharmaceutical composition may be a common dosage form. Also according to the present invention, telaprevir and camphor in the composition may be the same dosage form or different dosage forms.

[0109] Therefore, it should be understood that, according to the present invention, when the composition contains terapeptide and camphene...

[0110] When they are of the same dosage form, they can be administered simultaneously (i.e., at the same time) and using the same route of administration.

[0111] It should also be understood that, according to the present invention, when telaprevir and camphene in the composition are in the same or different dosage forms, they can be administered simultaneously, sequentially, or separately via the same or different routes of administration.

[0112] Preferably, when administered sequentially, terabhivir and camphor are administered at intervals not exceeding about 1 hour (preferably not exceeding about 10 minutes, even more preferably not exceeding about 1 minute).

[0113] According to the present invention, telaprevir and camphene can be mixed with one or more acceptable excipients or inert carriers (i.e., pharmaceutically inactive and non-toxic excipients) to impart a specific consistency or other specific physical or taste characteristics to the finished product, avoiding any chemical interaction with the two drugs. These include saline solutions, isotonic solutions, physiological solutions, buffer solutions, etc., which are compatible with the pharmaceutical use and known to those skilled in the art. The pharmaceutical compositions of the present invention may contain one or more reagents or media selected from the group consisting of: solubilizers, dispersants, stabilizers, sweeteners, preservatives, flavoring agents, lubricants, anti-caking agents, disintegrants, adsorbents, etc. In particular, reagents or media (liquid and / or injectable and / or solid) that can be used in formulations include: methylcellulose, carboxymethylcellulose, hydroxymethylcellulose, polysorbate 80, cyclodextrin, gelatin, mannitol, lactose, PEG, animal oils or vegetable oils, etc. The pharmaceutical compositions of the present invention can be formulated as follows: suspensions or ready-to-use or temporary injection solutions, oils, gels, tablets, powders, suppositories, capsules, granules, emulsions, suspensions, polymers, microspheres, nanoparticles, rectal capsules, enemas, pastes, creams, ointments, plasters, implants, decoctions, sprays, aerosols, etc.; optionally, controlled release and / or sustained release can be achieved through dosage forms or devices. For this type of formulation, reagents such as carbonates, cellulose, or starch are advantageously used.

[0114] In one embodiment, the pharmaceutical composition may be formulated into a powder form and reconstituted for intravenous injection.

[0115] Administration may be performed by any method known to those skilled in the art, preferably orally or parenterally, such as by intramuscular injection, intravenous administration, subcutaneous injection, intra-arterial injection, intraperitoneal injection, intracerebral injection or intrathecal injection, oral administration, sublingual administration, delivery to the lesion or brain or implantation, or aerosol administration. Intramuscular injection, intravenous administration, subcutaneous injection or oral administration are preferred. For long-term treatment, oral, sublingual or transdermal routes of administration are preferred.

[0116] For injection, the compound may be packaged as a suspension or liquid solution and administered using a syringe or infusion set. It should be understood that those skilled in the art can adjust the amount to be administered or the volume and / or rate of injection based on pathology, route of administration, individual circumstances, etc. It should be understood that repeated administration may be performed in combination with other active ingredients and / or any pharmaceutically acceptable carrier (buffer solution, isotonic solution, saline solution, in the presence of stabilizers, etc.).

[0117] According to some aspects, the pharmaceutical compositions of the present invention can be administered before, during, or after a process that may lead to cell death, such as before surgery, during surgery requiring bypassing the heart, or when an individual is at risk of potential ischemic injury (e.g., myocardial ischemia or vascular ischemia). According to some aspects, the pharmaceutical compositions of the present invention can be administered after a process that may lead to cell death has occurred (e.g., after an infarction).

[0118] In cases of pathological conditions and / or degenerative processes and / or cell death processes caused at least in part by ischemia-reperfusion, the compositions of the present invention may be administered before and / or during and / or after ischemia and / or before and / or during and / or after reperfusion.

[0119] Whether the organ is in situ or ex vivo, contact between the organ and the composition of the present invention can be made by any known means, such as direct contact with the organ by spraying, irrigation, immersion, dipping, rinsing, etc.

[0120] Optionally, as needed, the amount of the drug composition administered daily may be taken as one, two, three, four, five, six or more doses per day, or as multiple sub-dose taken at appropriate intervals throughout the day.

[0121] Optionally, the amount selected may depend on a variety of factors, particularly the route of administration, duration of administration, time of administration, rate of elimination of the compound, different products used in combination with the composition, the individual's age, weight and physical condition, as well as the individual's medical history, the nature of the pathological condition or degenerative process he or she is facing, and any other medically known information.

[0122] Optionally, the doctor may begin with a lower dose than normally used and then gradually increase these doses to better control potential side effects. Preferably, the compositions of the present invention can be administered for a period of 1 day to 20 years, and even more preferably for a period of 1 day to 3 years.

[0123] The present invention also relates to a treatment method that may include administering a therapeutically effective amount of a pharmaceutical composition comprising terabitvir and camphor to a person or animal in need.

[0124] The present invention also relates to a method for preparing a pharmaceutical composition comprising terabhivir and camphene, wherein the compound of the composition of the present invention is mixed with an acceptable excipient (particularly a pharmaceutically acceptable excipient) according to methods known per se.

[0125] The present invention also relates to the use of pharmaceutical compositions containing telaprevir and camphene in the preparation of organ preservation solutions. For example, organ preservation solutions containing pharmaceutical compositions containing telaprevir and camphene can be prepared from any existing organ preservation solution (e.g., any solution used for infusion, storage, transport, and / or flushing of organs). Such solutions may be, for example, Belzer cryopreservation solution, preservation solutions, or mixtures thereof.

[0126] Based on the same inventive concept, the present invention also provides: an organ preservation solution comprising the pharmaceutical composition described above.

[0127] Further, in the organ preservation solution, the concentration of telaprevir is from 0.01 mg / L to 1000 mg / L, preferably from 0.1 mg / L to 100 mg / L, more preferably from 1 mg / L to 10 mg / L; and the concentration of camphene is from 0.01 mg / L to 150 mg / L, preferably from 1 mg / L to 50 mg / L, more preferably from 1 mg / L to 10 mg / L.

[0128] Optionally, the organ preservation solution of the present invention can be used to inject the organ in situ before it is removed from the donor, optionally to cool it, and / or to use, for example, to statically flush and / or store and / or transport the organ after removal, or to inject it, for example, by using an infusion machine (with or without oxygen supply) at different temperatures ranging from low temperature to normal body temperature.

[0129] According to the present invention, the organ preservation solution contains an amount sufficient to prevent / mitigate / limit lesions caused by processes that can lead to cell death (particularly lesions caused by ischemia-reperfusion).

[0130] Advantageously, the organ preservation solution of the present invention can be used for infusion and / or storage and / or transportation and / or flushing of organs, such as the liver, lungs, heart, kidneys, or pancreas, preferably the liver. Optionally, the pharmaceutical composition of the present invention can be added to the solution several hours to minutes before using the organ preservation solution for infusion and / or storage and / or transportation and / or flushing of the organ.

[0131] Furthermore, the pharmaceutical composition can be added when the organ may already be present in the organ preservation solution. For example, the pharmaceutical composition can be added to the preservation solution at any time during warm ischemia, cold ischemia, or reperfusion.

[0132] Optionally, according to the present invention, when telaprevir and borneol in the pharmaceutical composition are of the same dosage form, they may be added simultaneously to the organ preservation solution. According to the present invention, when telaprevir and borneol in the pharmaceutical composition are of the same dosage form or different dosage forms, they may be added simultaneously or sequentially to the organ preservation solution.

[0133] Another object of the present invention relates to a method for temporarily preparing an organ preservation solution comprising telaprevir and camphene, the method comprising the step of mixing telaprevir and camphene. According to the present invention, when telaprevir and camphene are used to prepare a transplant organ preservation solution, the composition can be formulated into a form compatible with this use.

[0134] The present invention also relates to a method for preventing and / or protecting and / or treating organs, tissues or cells against ischemia-reperfusion injury (which may occur during actual warm or cold ischemia and / or actual reperfusion and / or ischemia-reperfusion), the method comprising contacting said organ, tissue or cells with an organ preservation solution of the composition of terabitrol and camphene of the present invention.

[0135] Another object of the present invention relates to the use of the telpivir and camphor composition, medicament, or preservation solution of the present invention in the prevention and / or protection and / or treatment of organs, tissues, or cells against ischemia-reperfusion injury (which may occur during actual warm or cold ischemia and / or actual reperfusion and / or ischemia-reperfusion).

[0136] Another object of the present invention relates to the use of the telpivir and camphor compositions, pharmaceuticals, compositions or preservative solutions of the present invention in the prevention and / or protection and / or treatment of organ, tissue or cell resistance to lesions that may occur during ischemia-reperfusion.

[0137] The use of camphor in the preparation of drugs for the treatment and / or prevention of multiple sclerosis.

[0138] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0139] The pharmaceutical composition of the present invention has excellent cell protection effect. The combined use of telaprevir and camphene shows a synergistic effect, which helps to achieve better efficacy, reduce drug dosage, improve clinical drug safety, and has good clinical application prospects.

[0140] The pharmaceutical compositions of the present invention can more effectively prevent and / or treat ischemia / reperfusion injury, especially nerve cell damage, and have excellent protective effects against ischemic stroke, significantly reducing cerebral ischemia / reperfusion injury. Attached Figure Description

[0141] Figure 1 Figure A shows the TTC staining and infarct volume measurement of mouse brain tissue from different groups in Example 1.1.

[0142] Figure 1 B is a graph showing the neurological function scores of mice in different groups in Example 1.1.

[0143] Figure 2 Figure A shows the infarct volume measurement of mouse brain tissue in different groups in Example 1.2.

[0144] Figure 2 B is a graph showing the neurological function scores of mice in different groups in Example 1.2.

[0145] Figure 3 Figure A shows the measurement of myocardial infarction area in mice of different groups in Example 3.

[0146] Figure 3 B is a graph showing the concentration of CK in the serum of mice in different groups in Example 3.

[0147] Figure 4 This is a graph showing the ratio of the time mice spent in the new arm in different groups in Example 5.

[0148] Figure 5 A is a diagram showing the escape latency of mice in different groups in Example 5.

[0149] Figure 5 B is a graph showing the ratio of time spent in the target quadrant by mice in different groups in Example 5.

[0150] Figure 6 This is a graph showing the time (latency) that mice in different groups spent on the rotarod in Example 7. Detailed Implementation

[0151] The present invention will now be described in detail with reference to the accompanying drawings and embodiments. It should be noted that, unless otherwise specified, the embodiments and features described herein can be combined with each other. The present invention will now be described in detail with reference to the embodiments.

[0152] Applications of telaprevir and camphene in the preparation of cell protection drugs.

[0153] Materials and methods:

[0154] To demonstrate the role of telaprevir and camphor in cell protection, the applicant used various animal models, such as ischemic stroke mouse model, myocardial ischemia / reperfusion injury mouse model, Alzheimer's disease, amyotrophic lateral sclerosis, multiple sclerosis, and Parkinson's disease animal models, and administered telaprevir and camphor at different time points. Cell damage and the protective effect of the drugs were detected according to the corresponding detection methods for each disease model.

[0155] Experimental reagents: telaprevir (compound) and dextroborneol (compound) were purchased from a reagent company and dissolved and prepared according to the company's reagent instructions.

[0156] Example 1

[0157] The role of pharmaceutical compositions containing telaprevir and camphor in the treatment of ischemic stroke:

[0158] Example 1.1

[0159] Animal experiments: Using a mouse model of ischemic stroke, the drug combination containing terabitvir and camphor was used to verify the anti-ischemic stroke effect.

[0160] Experimental animals: 7-week-old male C57BL / 6J mice. All experimental animals were housed in an SPF-grade housing for one week at a temperature of 22℃±2℃, relative humidity of 45%±15%, with free access to water and following a 12-hour light / dark cycle. Then, they were administered drugs according to the requirements of each experimental group.

[0161] Methods for establishing a mouse model of ischemic stroke: A mouse model of cerebral ischemia / reperfusion was established using the middle cerebral artery occlusion (MCAO) method. The steps are as follows: Eight-week-old male C57BL / 6J mice were anesthetized by intraperitoneal injection of 0.3% sodium pentobarbital (20 mL / kg). The left common carotid artery (CCA) was isolated, and the left external carotid artery (ECA) and internal carotid artery (ICA) were separated superiorly. The ECA and ICA were temporarily clamped with ophthalmic forceps, and the proximal end of the CCA was ligated. Then, a knotted spare suture was placed at the distal end of the CCA, a small incision was made at the lower end of the suture, and the suture was inserted into the internal carotid artery. The arterial clamps on the ECA and ICA were released, and the suture was advanced into the cranium along the ICA. The process stopped upon encountering resistance, the suture was tightened, and the suture was secured. After 1 hour of ischemia, the suture was removed, the skin was sutured, and the animals were treated after 24 hours of reperfusion.

[0162] The Longa 5-point scale was used to score the neurological deficits in the mouse model of cerebral ischemia / reperfusion injury (neurological function score). 0 points: no neurological deficit symptoms; 1 point: right forelimb cannot be fully extended; 2 points: rotation to the right; 3 points: walking towards the right and falling; 4 points: inability to walk spontaneously, loss of consciousness. Scores of 1-4 indicate a valid model.

[0163] TTC staining and infarct volume determination in mouse brains. After anesthetizing mice, the brains were quickly removed, the olfactory bulb and hindbrain were discarded, and 3-5 coronal slices were taken starting from the frontal pole. These slices were immediately placed in 1% TTC solution and incubated at 37°C in the dark for 30 minutes. Then, they were fixed by soaking in 10% paraformaldehyde solution. Infarcted areas appeared white, and non-infarcted areas appeared red. Each group of brain slices was neatly arranged and scanned. ImageJ was used to measure the infarct area and non-infarct area of ​​each brain slice, and the corresponding volume was calculated using the formula: Infarct volume (%) = (Volume of brain tissue contralateral to the infarct - Volume of brain tissue in the non-infarcted area on the infarcted side) / Volume of brain tissue contralateral to the infarct × 100%.

[0164] Experimental grouping and administration: Experimental animals were randomly divided into the following groups, with 6 animals in each group. Telapirvir, dexborneol, and other related drugs were dissolved in the solvent (10% DMSO + 30% PEG400 + 60% physiological saline), i.e.:

[0165] Sham group: The internal and external carotid arteries were separated without inserting sutures into the arteries.

[0166] Cerebral ischemia / reperfusion group (I / R group): cerebral ischemia for 1 hour and reperfusion for 24 hours were performed.

[0167] Telaprevir + I / R group: During the above reperfusion period, telaprevir (30 mg / kg, i.e., 30 mg of telaprevir per 1 kg of mouse body weight) was administered intramuscularly 1 hour after reperfusion, and the results were obtained 24 hours after reperfusion.

[0168] Borneol + I / R group: During the above reperfusion period, dexborneol (1 mg / kg) was administered intramuscularly 1 hour after reperfusion, and the results were measured 24 hours after reperfusion.

[0169] Telaprevir + dexborneol + cerebral ischemia / reperfusion group (Telaprevir + Borneol + I / R): During the above reperfusion period, telaprevir (30 mg / kg) and dexborneol (1 mg / kg) were administered intramuscularly 1 hour after reperfusion, and the results were obtained 24 hours after reperfusion.

[0170] Telaprevir + dexborneol + cerebral ischemia / reperfusion group (Telaprevir + Borneol + I / R): During the above reperfusion period, telaprevir (30 mg / kg) and dexborneol (0.75 mg / kg) were administered intramuscularly 1 hour after reperfusion, and the results were measured 24 hours after reperfusion.

[0171] The solvent + cerebral ischemia / reperfusion group (Vehicle + I / R) was tested 24 hours after reperfusion, following intramuscular injection of solvent during the reperfusion period.

[0172] The solvent is 10% DMSO + 30% PEG400 + 60% physiological saline (i.e., composed of DMSO, PEG400, and physiological saline in a volume ratio of 1:3:6); terabitvir and / or dextromethorphan are dissolved in the solvent and then used for injection (the same applies below).

[0173] The neurological function scores of mice and the volume of cerebral infarction were measured to evaluate the drug's efficacy.

[0174] Statistical analysis: Statistical analysis was performed using GraphPad Prism 9.0 software. All data are expressed as mean ± standard error (±SEM). Differences among multiple groups were analyzed using one-way ANOVA, followed by Tukey's test to compare whether there were statistically significant differences between groups. Neurological function scores were analyzed using Kruskal-Wallis and Wilcoxon tests. A p-value < 0.05 was considered statistically significant (the same applies below).

[0175] result:

[0176] Effects of the combination of terabhivir and dextroborneol on cerebral infarction volume and neurological function in mice

[0177] like Figure 1 As shown in the figure, the I / R group had obvious white infarct foci, while the terabhivir, dexborneol monotherapy group, and the terabhivir and dexborneol combination therapy group significantly reduced the infarct volume in mice (A) and improved neurological function (B). The dexborneol monotherapy group had no significant effect, while the combination therapy was significantly better than the monotherapy. (Data are expressed as mean ± standard error, n = 6, **P < 0.01 vs sham operation group;) # P<0.05 ## P<0.01 vs I / R group; + P<0.05 vs I / R + drug alone group

[0178] Conclusion: The combined use of telaprevir and dexborneol is more effective than either drug, exhibiting a synergistic effect. It can significantly reduce neuronal cell death, alleviate cerebral ischemia / reperfusion injury, and has a neuroprotective effect. The drug combination of telaprevir and dexborneol can be used to prepare drugs to alleviate cerebral ischemia / reperfusion injury for the treatment of ischemic stroke.

[0179] Example 1.2

[0180] Animal experiments: Using a mouse model of ischemic stroke, the drug combination containing terabitvir and camphor was used to verify the anti-ischemic stroke effect.

[0181] Experimental animals: 7-week-old male C57BL / 6J mice. All experimental animals were housed in an SPF-grade housing for one week at a temperature of 22℃±2℃, relative humidity of 45%±15%, with free access to water and following a 12-hour light / dark cycle. Then, they were administered drugs according to the requirements of each experimental group.

[0182] Methods for establishing a mouse model of ischemic stroke: A mouse model of cerebral ischemia / reperfusion was established using the middle cerebral artery occlusion (MCAO) method. The steps are as follows: Eight-week-old male C57BL / 6J mice were anesthetized by intraperitoneal injection of 0.3% sodium pentobarbital (20 mL / kg). The left common carotid artery (CCA) was isolated, and the left external carotid artery (ECA) and internal carotid artery (ICA) were separated superiorly. The ECA and ICA were temporarily clamped with ophthalmic forceps, and the proximal end of the CCA was ligated. Then, a knotted spare suture was placed at the distal end of the CCA, a small incision was made at the lower end of the suture, and the suture was inserted into the internal carotid artery. The arterial clamps on the ECA and ICA were released, and the suture was advanced into the cranium along the ICA. The process stopped upon encountering resistance, the suture was tightened, and the suture was secured. After 1 hour of ischemia, the suture was removed, the skin was sutured, and the animals were treated after 24 hours of reperfusion.

[0183] The Longa 5-point scale was used to score the neurological deficits in the mouse model of cerebral ischemia / reperfusion injury (neurological function score). 0 points: no neurological deficit symptoms; 1 point: right forelimb cannot be fully extended; 2 points: rotation to the right; 3 points: walking towards the right and falling; 4 points: inability to walk spontaneously, loss of consciousness. Scores of 1-4 indicate a valid model.

[0184] TTC staining and infarct volume determination in mouse brains. After anesthetizing mice, the brains were quickly removed, the olfactory bulb and hindbrain were discarded, and 3-5 coronal slices were taken starting from the frontal pole. These slices were immediately placed in 1% TTC solution and incubated at 37°C in the dark for 30 minutes. Then, they were fixed by soaking in 10% paraformaldehyde solution. Infarcted areas appeared white, and non-infarcted areas appeared red. Each group of brain slices was neatly arranged and scanned. ImageJ was used to measure the infarct area and non-infarct area of ​​each brain slice, and the corresponding volume was calculated using the formula: Infarct volume (%) = (Volume of brain tissue contralateral to the infarct - Volume of brain tissue in the non-infarcted area on the infarcted side) / Volume of brain tissue contralateral to the infarct × 100%.

[0185] Depending on the administration method and dosage, the experiment was divided into two parts: the neurological function score of mice was measured and the volume of cerebral infarction was measured to evaluate the drug effect.

[0186] Statistical analysis: Statistical analysis was performed using GraphPad Prism 9.0. All data are expressed as mean ± standard error (±SEM). Differences among multiple groups were analyzed using one-way ANOVA, followed by Tukey's test to compare whether there were significant differences between groups. Neurological function scores were analyzed using Kruskal-Wallis and Wilcoxon tests. P < 0.05 was considered statistically significant (the same applies below).

[0187] Experimental grouping and administration method: The experimental animals were randomly divided into the following groups, with 6 animals in each group. Telapirvir and dextroborneol were dissolved in a solvent [0.5% (w / v) aqueous solution of sodium carboxymethyl cellulose], i.e.:

[0188] Sham group: The internal and external carotid arteries were separated without obstructing the arteries.

[0189] Cerebral ischemia / reperfusion group (I / R group): cerebral ischemia for 1 hour and reperfusion for 24 hours were performed.

[0190] Cerebral ischemia / reperfusion + low-dose terabitvir group [+terabitvir L (Low)]: During the above reperfusion period, terabitvir (50 mg / kg) was administered by gavage 1 h and 5 h after reperfusion, and the results were measured 24 h after reperfusion.

[0191] Cerebral ischemia / reperfusion + high-dose terabitvir group [+terabitvir H (High)]: During the above reperfusion period, terabitvir (100 mg / kg) was administered by gavage 1 h and 5 h after reperfusion, and the results were measured 24 h after reperfusion.

[0192] Cerebral ischemia / reperfusion + low-dose dexborneol group (+ dexborneol L): During the above reperfusion period, dexborneol (2.5 mg / kg) was administered by gavage 1 h and 5 h after reperfusion, and the results were measured 24 h after reperfusion.

[0193] Cerebral ischemia / reperfusion + high-dose dexborneol group (+dexborneol H): During the above reperfusion period, dexborneol (3.5 mg / kg) was administered by gavage 1 h and 5 h after reperfusion, and the results were measured 24 h after reperfusion.

[0194] Cerebral ischemia / reperfusion + low-dose terabhivir and high-dose dexborneol combination group 1 (+composition 1): During the above reperfusion period, 1 hour and 5 hours after reperfusion, composition 1 (the composition of composition 1 satisfies the following: after each administration of composition 1, the current dose of terabhivir is 50 mg / kg and the current dose of dexborneol is 3.5 mg / kg), i.e., T50+B3.5 (where T represents terabhivir, B represents dexborneol, and the numbers are the corresponding drug dosages, with the units of the numbers being mg / kg) were administered by gavage, and the results were measured 24 hours after reperfusion.

[0195] Cerebral ischemia / reperfusion + high-dose terabitrin and low-dose dexborneol combination group 2 (+composition 2): During the above reperfusion period, composition 2 was administered 1 h and 5 h after reperfusion (the composition of composition 2 satisfies: after each administration of composition 2, the current dose of terabitrin is 100 mg / kg and the current dose of dexborneol is 2.5 mg / kg), i.e., T100+B2.5 by gavage, and the results were measured 24 h after reperfusion.

[0196] Cerebral ischemia / reperfusion + solvent group (+ solvent): During the above reperfusion period, solvent was administered by gavage 1 h and 5 h after reperfusion. The solvent was 0.5% (w / v) sodium carboxymethyl cellulose aqueous solution.

[0197] The neurological function scores of mice were assessed and the volume of cerebral infarction was measured to evaluate the drug's efficacy. Experimental results:

[0198] Effects of the combination of terabhivir and dextroborneol on cerebral infarction volume and neurological function in mice.

[0199] like Figure 2 As shown, the infarct volume in mice in the cerebral ischemia / reperfusion group and the solvent group was significantly increased. Oral administration of telaprevir monotherapy (100 mg / kg) and drug combinations containing telaprevir and dextroborneol (T50 mg / kg + B3.5 mg / kg, T100 mg / kg + B2.5 mg / kg) significantly reduced the infarct volume in mice. Figure 2 A) and improving neurological function ( Figure 2 B). Compared with terabhidimib and dexborneol monotherapy, the drug combination containing terabhidimib and dexborneol (T50 mg / kg + B3.5 mg / kg, T100 mg / kg + B2.5 mg / kg) was significantly more effective than terabhidimib and dexborneol monotherapy. Therefore, the drug combination containing terabhidimib and dexborneol can significantly reduce cerebral ischemia / reperfusion injury and has a therapeutic effect on ischemic stroke, which is better than the effect of terabhidimib and dexborneol monotherapy, showing a synergistic effect. (Data are expressed as mean ± standard error, number of mice in each group n = 6, **P < 0.01 vs sham-operated group;) # P<0.05 ## P<0.01 vs I / R group; + P<0.05 ++ P<0.01 vs I / R + drug alone group).

[0200] Conclusion: The pharmaceutical composition containing telaprevir and dexborneol of the present invention has a better therapeutic effect than single drugs, exhibiting a synergistic effect. It can significantly reduce neuronal cell death, alleviate cerebral ischemia / reperfusion injury, and has a neuroprotective effect. The pharmaceutical composition of telaprevir and dexborneol can be used to prepare drugs to alleviate cerebral ischemia / reperfusion injury for the treatment of ischemic stroke.

[0201] The above examples illustrate that the pharmaceutical composition containing telaprevir and dexborneol can be used to treat stroke (including ischemic stroke), and that the combined use of telaprevir and dexborneol is more effective than single drugs, showing a synergistic effect, which helps to ensure efficacy and reduce drug dosage, thereby improving the safety of clinical use while ensuring clinical treatment effect.

[0202] However, this invention is not limited to the above-mentioned diseases; the drug is also applicable to the treatment of other diseases with similar pathogenesis.

[0203] Example 2

[0204] Validating the anti-myocardial ischemia / reperfusion injury effect of a pharmaceutical composition containing telaprevir and camphor.

[0205] Animal experiments: Using a mouse model of myocardial ischemia / reperfusion injury, the effects of a drug composition containing telaprevir and camphor on myocardial ischemia / reperfusion injury were verified.

[0206] Experimental animals: 7-week-old male C57BL / 6J mice. All experimental animals were housed in an SPF-grade housing for one week at a temperature of 22℃±2℃, relative humidity of 45%±15%, with free access to water and following a 12-hour light / dark cycle. Then, they were administered drugs according to the requirements of each experimental group.

[0207] Methods for establishing a mouse model of myocardial ischemia / reperfusion: Eight-week-old male C57BL / 6J mice were anesthetized by intraperitoneal injection of 0.3% sodium pentobarbital (20 mL / kg), followed by endotracheal intubation, with the endotracheal tube fixed using pressure-sensitive adhesive. Ventilator parameters were set to a tidal volume of 3.2 mL / kg and a frequency of 110 breaths per minute. The left anterior descending artery (LAD) was ligated using 8-0 sutures after thoracotomy. Myocardial infarction was confirmed by whitening at the apex of the heart, and the ligation was maintained for 1 hour. After 1 hour of ischemia, the LAD suture knot was opened to restore cardiac perfusion. The thoracic cavity was sutured layer by layer, and the ventilator was removed; the mice regained spontaneous breathing. Twenty-four hours after cardiac perfusion was restored, the mice were anesthetized again with 0.3% sodium pentobarbital (20 mL / kg), and the LAD was ligated again in situ after thoracotomy. The abdominal cavity was opened, and 0.2 mL of 2% Evans Blue solution was injected via the inferior vena cava. When the lower lip of the mouse turned blue, blood was collected from the apex of the heart, and the heart was removed. The heart was frozen at -20°C for 1 hour, then sliced ​​into 1 mm thick sections. The sections were then incubated with 1% TTC staining solution (prepared in PBS) at 37°C for 15 minutes. After staining, the staining solution was removed, the sections were washed once with PBS, and fixed with 4% paraformaldehyde for 24 hours. The heart sections were then removed, the staining was observed, and photographs were taken. The areas of ischemic and infarcted regions were determined using ImageJ software.

[0208] The blue area in the heart slice represents normal tissue; the area outside the blue area is the ischemic area (also known as the risk area, AOR); the white area is the infarct area (AOI). The percentage of infarct area in each slice (the ratio of the infarct area AOI to the ischemic area AOR) is calculated, and the ratio of the infarct area is compared between the drug group and the model group.

[0209] Serum creatine kinase (CK) activity assay: After ischemia / reperfusion surgery, approximately 150 μL of whole blood was collected from the orbit of mice. The blood was centrifuged at 3000 rpm for 10 min at 4°C, and the supernatant was collected and stored at -40°C. Serum CK activity was measured according to the instructions of a commercially available kit as follows: 10 mL of R2 was dissolved in one vial of R1 to prepare the working solution. 4 μL of serum was added to 200 μL of the CK kit working solution. The solution was incubated at 37°C for 2 min. The wavelength was set to 340 nm using a microplate reader. Absorbance readings (A0, A1, A2, A3) were taken at 0, 1, 2, and 3 min. The average change in absorbance per minute (ΔA) was calculated, and the concentration of CK in the serum (U / L) was calculated.

[0210] Experimental grouping: Experimental animals were randomly divided into groups of 6-10. Based on preliminary experimental results, drug concentrations and administration times were set. The drug groups were treated with different concentrations of terabitvir, camphor, and a combination of both drugs, respectively.

[0211] Sham group: The mice underwent heart surgery but no vascular ligation was performed;

[0212] Model group (myocardial ischemia / reperfusion group): Ligation of the left anterior descending coronary artery for 1 hour to treat ischemia, suture cut, and reperfusion for 24 hours; telaprevir +

[0213] Myocardial ischemia / reperfusion group (Telaprevir+I / R): mice were treated with teraprevir 30 min after ischemia.

[0214] Borneol + myocardial ischemia / reperfusion group (Borneol+I / R): mice were treated with borneol 30 min after ischemia.

[0215] Telaprevir + Borneol + I / R: Mice were treated with telaprevir and borneol 30 min after ischemia.

[0216] The solvent + myocardial ischemia / reperfusion group (Vehicle + I / R) was treated with solvent after mice were ischemic for 30 min.

[0217] Blood and myocardial tissue were collected and relevant indicators were measured: myocardial infarction area was measured in mice and serum creatine kinase activity (CK activity) was detected.

[0218] Results: Telapir or camphor monotherapy and the combination of the two drugs have the effect of resisting myocardial ischemia / reperfusion injury in mice. They can reduce the infarct area of ​​myocardial tissue, reduce serum creatine kinase, and reduce myocardial cell death, thus having a cardioprotective effect. The effect of combination therapy is significantly better than that of monotherapy.

[0219] Conclusion: The combined use of telaprevir and camphor is more effective than single-drug therapy, exhibiting a synergistic effect. It can significantly reduce cardiomyocyte death, alleviate myocardial ischemia / reperfusion injury, and has a cardioprotective effect. It can be used to prepare drugs to alleviate myocardial ischemia / reperfusion injury for the treatment of myocardial infarction.

[0220] Example 3

[0221] Validating the anti-myocardial ischemia / reperfusion injury effect of a pharmaceutical composition containing telaprevir and camphor.

[0222] Animal experiments: Using a mouse model of myocardial ischemia / reperfusion injury, the effects of a drug composition containing telaprevir and camphor on myocardial ischemia / reperfusion injury were verified.

[0223] Experimental animals: 7-week-old male C57BL / 6J mice. All experimental animals were housed in an SPF-grade housing for one week at a temperature of 22℃±2℃, relative humidity of 45%±15%, with free access to water and following a 12-hour light / dark cycle. Then, they were administered drugs according to the requirements of each experimental group.

[0224] Methods for establishing a mouse model of myocardial ischemia / reperfusion: Eight-week-old male C57BL / 6J mice were anesthetized by intraperitoneal injection of 0.3% sodium pentobarbital (20 mL / kg), followed by endotracheal intubation, with the endotracheal tube fixed using pressure-sensitive adhesive. Ventilator parameters were set to a tidal volume of 3.2 mL / kg and a frequency of 110 breaths per minute. The left anterior descending artery (LAD) was ligated using 8-0 sutures after thoracotomy. Myocardial infarction was confirmed by whitening at the apex of the heart, and the ligation was maintained for 1 hour. After 1 hour of ischemia, the LAD suture knot was loosened to restore cardiac perfusion. The thoracic cavity was sutured layer by layer, and the ventilator was removed; the mice regained spontaneous breathing. Twenty-four hours after cardiac perfusion was restored, the mice were anesthetized again with 0.3% sodium pentobarbital (20 mL / kg), and the LAD was ligated again in situ after thoracotomy. The abdominal cavity was opened, and 0.2 mL of 2% Evans Blue solution was injected via the inferior vena cava. When the lower lip of the mouse turned blue, blood was collected from the apex of the heart, and the heart was removed. The heart was frozen at -20°C for 1 hour, then sliced ​​into 1 mm thick sections. The sections were then incubated with 1% TTC staining solution (prepared in PBS) at 37°C for 15 minutes. After staining, the staining solution was removed, the heart was washed once with PBS, and fixed with 4% paraformaldehyde for 24 hours. The heart sections were then removed, the staining was observed, and photographs were taken. The areas of ischemic and infarcted regions were determined using ImageJ software.

[0225] The blue area in the heart slice represents normal tissue; the area outside the blue area is the ischemic area (also known as the risk area, AOR); the white area is the infarct area (AOI). The percentage of infarct area in each slice (the ratio of the infarct area AOI to the ischemic area AOR) is calculated, and the ratio of the infarct area is compared between the drug group and the model group.

[0226] Serum creatine kinase (CK) activity assay: After ischemia / reperfusion surgery, approximately 150 μL of whole blood was collected from the orbit of mice. The blood was centrifuged at 3000 rpm for 10 min at 4°C, and the supernatant was collected and stored at -40°C. Serum CK activity was measured according to the instructions of a commercially available kit as follows: 10 mL of R2 was dissolved in one vial of R1 to prepare the working solution. 4 μL of serum was added to 200 μL of the CK kit working solution. The solution was incubated at 37°C for 2 min. The wavelength was set to 340 nm using a microplate reader. Absorbance readings (A0, A1, A2, A3) were taken at 0, 1, 2, and 3 min. The average change in absorbance per minute (ΔA) was calculated, and the concentration of CK in the serum (U / L) was calculated.

[0227] Experimental grouping: Experimental animals were randomly divided into groups of 6. Based on preliminary experimental results, drug concentrations and administration times were set. The drug groups were treated with different concentrations of terabhivir, dextroborneol, and a combination of both drugs. The solvent was 10% DMSO + 30% PEG400 + 60% physiological saline (i.e., composed of DMSO, PEG400, and physiological saline in a 1:3:6 volume ratio). Terabhivir and / or dextroborneol were dissolved in the solvent and then injected.

[0228] Sham surgery group: The hearts of mice were operated on, but blood vessels were not ligated;

[0229] Model group (myocardial ischemia / reperfusion group, I / R group): left anterior descending coronary artery was ligated for 1 hour for ischemia treatment, the suture was cut, and reperfusion was performed for 24 hours;

[0230] Myocardial ischemia / reperfusion + low-dose terabhivir group (+terabhivir L): terabhivir (7.5 mg / kg) was administered intramuscularly to mice 30 min after ischemia and 5 h after reperfusion, and the results were measured 24 h after reperfusion.

[0231] Myocardial ischemia / reperfusion + high-dose terabitvir group (+terabitvir H): terabitvir (15 mg / kg) was administered intramuscularly to mice 30 min after ischemia and 5 h after reperfusion, and the results were measured 24 h after reperfusion.

[0232] Myocardial ischemia / reperfusion + dexborneol group (+ dexborneol): Mice were given dexborneol (0.75 mg / kg) intramuscularly 30 min after ischemia and 5 h after reperfusion, and the results were measured 24 h after reperfusion.

[0233] Myocardial ischemia / reperfusion + low-dose terabhivir and dexborneol combination group 1 (+composition 1): Mice were given intramuscular injection of composition 1 (the composition of composition 1 satisfies the following conditions: 7.5 mg / kg of terabhivir and 0.75 mg / kg of dexborneol after each administration) 30 min after ischemia and 5 h after reperfusion. The results were measured 24 h after reperfusion.

[0234] Cerebral ischemia / reperfusion + high-dose terabhivir and dexborneol combination group 2 (+composition 2): After 30 min of ischemia and 5 h of reperfusion in mice, composition 2 (the composition of composition 2 satisfies the following: after each administration, the dose of terabhivir is 15 mg / kg and the dose of dexborneol is 0.75 mg / kg) was administered intramuscularly, and the results were measured 24 h after reperfusion.

[0235] Myocardial ischemia / reperfusion + solvent group (+ solvent): During the above reperfusion period, solvent was injected intramuscularly 1 hour and 5 hours after reperfusion, and the results were measured 24 hours after reperfusion.

[0236] Blood and myocardial tissue were collected and relevant indicators were measured: myocardial infarction area was measured in mice and serum creatine kinase activity (CK activity) was detected.

[0237] Statistical analysis: Statistical analysis was performed using GraphPad Prism 9.0 software. All data are expressed as mean ± standard error (±SEM). Independent samples Student's t-test was used to compare means between two groups, and one-way ANOVA was used to analyze differences among multiple groups. Tukey's test was used for post-hoc multiple comparisons. A p-value < 0.05 was considered statistically significant.

[0238] Result: As Figure 3 As shown, the infarct area of ​​myocardial tissue in mice in the I / R group and the solvent group was significantly increased. Intramuscular injection of terabitvir monotherapy (15 mg / kg) and drug combinations containing terabitvir and dextroborneol (T7.5 mg / kg + B0.75 mg / kg, T15 mg / kg + B0.75 mg / kg) significantly reduced the infarct area of ​​myocardial tissue in mice. Figure 3 A) Reduce serum creatine kinase ( Figure 3B). Compared with terabhidimib and dexborneol alone, the drug combination containing terabhidimib and dexborneol (T7.5 mg / kg + B0.75 mg / kg, T15 mg / kg + B0.75 mg / kg) was significantly more effective than terabhidimib and dexborneol alone. Therefore, the drug combination containing terabhidimib and dexborneol can significantly reduce myocardial ischemia / reperfusion injury and has a therapeutic effect on myocardial infarction. Its effect is better than that of terabhidimib and dexborneol alone, showing a synergistic effect. (Data are expressed as mean ± standard error, number of mice in each group n = 6, **P < 0.01 vs sham-operated group;) # P<0.05 ## P<0.01 vs I / R group; + P<0.05 ++ P<0.01 vs I / R + drug alone group).

[0239] Conclusion: The pharmaceutical composition containing telaprevir and dexborneol of the present invention has a better therapeutic effect than single drugs, exhibiting a synergistic effect. It can significantly reduce cardiomyocyte death, alleviate myocardial ischemia / reperfusion injury, and has a cardiomyocyte protective effect. The pharmaceutical composition of telaprevir and dexborneol can be used to prepare drugs to alleviate myocardial ischemia / reperfusion injury for the treatment of myocardial infarction.

[0240] Example 4

[0241] Verify the anti-Alzheimer's disease effect of a drug composition containing telaprevir and camphor.

[0242] Animal experiments: using Aβ 1-42 Animal models of Alzheimer's disease or APP / PS1 transgenic mice were constructed to verify the anti-Alzheimer's disease effect of drug compositions containing terabitvir and camphene.

[0243] Laboratory animals: APP / PS1 transgenic mice (Alzheimer's disease animal model, with C57BL / 6J mice as the background), male, 5 months old, weighing 24–30 g, and age-matched C57BL / 6J mice (male, 5 months old, weighing 26–30 g) were purchased from Beijing Huafukang Biotechnology Co., Ltd. All laboratory animals were housed in an SPF-grade enclosure with a temperature of 22℃±2℃, relative humidity of 45%±15%, free access to water, and following a 12-hour light / dark cycle.

[0244] Experimental grouping: Experimental animals were randomly divided into groups of 6-10. Based on preliminary experimental results, drug concentrations and administration times were set. The drug groups were treated with different concentrations of terabitvir, camphor, and a combination of both drugs.

[0245] Normal control group: APP / PS1 transgenic background mice, such as C57BL / 6J mice, plus intramuscular injection of solvent.

[0246] APP / PS1+solvent group: APP / PS1 mice + solvent.

[0247] APP / PS1 + Telaprevir group: APP / PS1 mice treated with Telaprevir.

[0248] APP / PS1 + Boneol group: APP / PS1 mice treated with camphor.

[0249] APP / PS1 + teraprevir group + camphor (+Telaprevir+Borneol): APP / PS1 mice were treated with teraprevir and camphor.

[0250] The learning and memory abilities and cognitive functions of mice were tested using experiments such as the new object recognition test and the Morris water maze test.

[0251] Detection methods and results:

[0252] (1) Effects of telaprevir and camphor combination on non-spatial learning and memory ability and cognitive function in Alzheimer's disease model mice - new object recognition experiment.

[0253] The experiment was conducted according to the requirements for new object recognition, and included three phases: adaptation, training, and testing. For one week prior to the experiment, the mice were petted for 2-3 minutes daily to reduce stress. A 40×40cm open-field test chamber was used. Before the experiment, the mice were placed in the test chamber to acclimatize for 20-30 minutes. After acclimatization, the new object training experiment began. Two objects (A and B) of identical color, shape, and material were placed in the test chamber, and the mice were placed in it for 10 minutes of training. Twenty-four hours after the training period, a new object testing experiment was conducted to assess the mice's short-term non-spatial learning memory. One of the two objects was replaced with another object of different shape and color (C), and the time taken for the mice to explore the two different objects within 10 minutes was recorded. Cognitive index = (New object exploration time / (New object exploration time + Old object exploration time)) × 100%.

[0254] Experimental results: Compared with normal control mice, the cognitive index of APP / PS1 + solvent group mice was significantly reduced. The non-spatial learning and memory ability and cognitive function of APP / PS1 mice were weakened. After administration of terabitvir, camphor monotherapy and the combination of the two drugs, the cognitive index of APP / PS1 mice was significantly increased. The combination of the two drugs was more effective than the monotherapy.

[0255] Conclusion: The combination of telaprevir and camphor significantly improves non-spatial learning and memory abilities and cognitive function in Alzheimer's disease mice.

[0256] (2) Effects of telaprevir and camphor combination on spatial learning and memory ability and cognitive function in Alzheimer's disease model mice—Morris water maze test.

[0257] Following the requirements of the water maze experiment, the Morris water maze was divided into four quadrants. A platform with a diameter of 12 cm and a height of 35 cm (the target quadrant) was placed in one quadrant, submerged in water by 1-2 cm, and the water was dyed black with carbon ink. After five days of orientation and navigation experiments, the platform was removed, and a spatial exploration experiment was conducted. The time it took for the mice to reach the original platform location (the latency period) and the time spent in the target quadrant (the time spent in the quadrant where the original hidden platform was located) were recorded, and the percentage of time spent in the target quadrant relative to the total exploration time was analyzed. The animals' movement trajectories and behaviors were recorded using the Smart 3.0 small animal analysis software system, and the relevant data were analyzed.

[0258] Compared with normal control mice, the APP / PS1+ solvent group mice had a significantly increased time to find the original platform location (escape latency), and a significantly reduced time spent in the target quadrant and the ratio of time spent in the target quadrant. After administration of terabitvir, or camphor monotherapy, or the combination of the two drugs, the APP / PS1 mice had a significantly reduced time to find the original platform (escape latency), and a significantly increased time spent in the target quadrant and the ratio of time spent in the target quadrant. The combination of the two drugs was more effective than the monotherapy.

[0259] Conclusion: The combination of telaprevir and camphor significantly improved the learning and memory abilities and cognitive functions of Alzheimer's disease mice, demonstrating a synergistic effect.

[0260] Example 5

[0261] Verify the anti-Alzheimer's disease effect of a drug composition containing telaprevir and camphor.

[0262] Animal experiments: using Aβ 1-42 Animal models of Alzheimer's disease or APP / PS1 transgenic mice were constructed to verify the anti-Alzheimer's disease effect of drug compositions containing terabitvir and camphene.

[0263] Laboratory animals: APP / PS1 transgenic mice (Alzheimer's disease animal model, with C57BL / 6J mice as the background), male, 5 months old, weighing 24–30 g, and age-matched C57BL / 6J mice (male, 5 months old, weighing 26–30 g) were purchased from Beijing Huafukang Biotechnology Co., Ltd. All laboratory animals were housed in an SPF-grade enclosure with a temperature of 22℃±2℃, relative humidity of 45%±15%, free access to water, and following a 12-hour light / dark cycle.

[0264] Experimental grouping: Five-month-old APP / PS1 transgenic mice were randomly divided into groups of six. Based on preliminary experimental results, drug concentrations and administration times were set. The drug groups were treated with different concentrations of telaprevir, dextroborneol, and a combination of both drugs, starting at 5 months of age via gavage, once daily for two consecutive months. The normal control group and the APP / PS1 mouse control group were given the same volume of solvent [0.5% (w / v) sodium carboxymethyl cellulose aqueous solution]. Specific groupings are as follows:

[0265] Normal control group: APP / PS1 transgenic background mice, such as C57BL / 6J mice, were administered solvent by gavage.

[0266] APP / PS1+solvent group: APP / PS1 mice were administered solvent by gavage.

[0267] APP / PS1 + low-dose terabhivir group (+terabhivir L): APP / PS1 mice were administered terabhivir (50 mg / kg) by gavage.

[0268] APP / PS1 + high-dose terabhivir group (+terabhivir H): APP / PS1 mice were administered terabhivir (100 mg / kg) by gavage.

[0269] APP / PS1 + dexborneol group (+ dexborneol): APP / PS1 mice were administered dexborneol (2.5 mg / kg) by gavage.

[0270] APP / PS1 + low-dose terabhivir and dexborneol combination group 1 (+composition 1): APP / PS1 mice + composition 1 (the composition of composition 1 satisfies the following: the current dose of terabhivir is 50 mg / kg and the current dose of dexborneol is 2.5 mg / kg after each administration) were administered by gavage.

[0271] APP / PS1 + high-dose terabhivir and dexborneol combination 2 group (+composition 2): APP / PS1 mice + composition 2 (the composition of composition 2 satisfies: after each administration, the current dose of terabhivir is 100 mg / kg and the current dose of dexborneol is 2.5 mg / kg) were administered by gavage.

[0272] The learning and memory abilities and cognitive functions of mice were tested using experiments such as the new object recognition test, the Y maze, and the Morris water maze.

[0273] Statistical analysis: GraphPad Prism 9.0 software was used for statistical analysis. All data are expressed as mean ± standard error (±SEM). Independent samples Student's t-test was used to compare the means of two groups. One-way ANOVA was used to analyze differences among multiple groups. Tukey's test was used for post-hoc multiple comparisons. P < 0.05 was considered statistically significant.

[0274] Detection methods and results:

[0275] (1) Effects of telaprevir and camphor combination on non-spatial learning and memory ability and cognitive function in Alzheimer's disease model mice - new object recognition experiment.

[0276] The experiment was conducted according to the requirements for new object recognition, and included three phases: adaptation, training, and testing. For one week prior to the experiment, the mice were petted for 2-3 minutes daily to reduce stress. A 40×40cm open-field test chamber was used. Before the experiment, the mice were placed in the test chamber to acclimatize for 20-30 minutes. After acclimatization, the new object training experiment began. Two objects (A and B) of identical color, shape, and material were placed in the test chamber, and the mice were placed in it for 10 minutes of training. Twenty-four hours after the training period, a new object testing experiment was conducted to assess the mice's short-term non-spatial learning memory. One of the two objects was replaced with another object of different shape and color (C), and the time taken for the mice to explore the two different objects within 10 minutes was recorded. Cognitive index = (New object exploration time / (New object exploration time + Old object exploration time)) × 100%.

[0277] Experimental results: Compared with normal control mice, the cognitive index of APP / PS1 + solvent group mice was significantly reduced. The non-spatial learning and memory ability and cognitive function of APP / PS1 mice were weakened. After administration of terabitvir, camphor monotherapy and the combination of the two drugs, the cognitive index of APP / PS1 mice was significantly increased. The combination of the two drugs was more effective than the monotherapy.

[0278] Conclusion: The combination of telaprevir and camphor significantly improves non-spatial learning and memory abilities and cognitive function in Alzheimer's disease mice.

[0279] (2) Y-maze experiment

[0280] This experiment was conducted according to reference 1 to test the learning and memory abilities of mice. The Y-shaped maze used in the experiment was made of gray organic plastic and consisted of three arms with an angle of 120° between adjacent arms. Each arm was 8cm × 30cm × 15cm (width × length × height). The computer could automatically track and record the mouse's exploration process using a camera located above the center of the maze. Three identical arms were randomly assigned: (1) the starting arm, which the mouse began to explore (always open); (2) the novel arm (referred to as "new arm"), which was blocked during the first test but opened during the second test; and (3) the other arms (always open). The novel arm was closed, and the mouse was placed in the starting arm with its back to the center of the maze, allowing it to explore orient itself in the starting arm and the other arms for 3 minutes. After the orienting exploration was completed, the mouse was removed from the Y maze and allowed to rest for 2 minutes. During this time, the maze was wiped with alcohol to remove the influence of the mouse's odor. After 2 minutes, the novel arm was opened, and the mouse was placed in the starting arm with its back to the center of the maze, allowing it to explore freely in the three arms for 1 minute. The movement trajectory and behavior of the animals were recorded using the Smart 3.0 small animal analysis software system. The time the mice spent on the "novel arm" was recorded, and the percentage of time the mice spent on the novel arm relative to the total free exploration time was analyzed (i.e., the novel arm time ratio).

[0281] Experimental results are as follows Figure 4 As shown in the figure, compared with the normal control group, the APP / PS1 + solvent group mice had a significantly shorter time spent on the "novel arm," indicating that the learning and memory abilities and cognitive functions of APP / PS1 mice were weakened. Administration of low and high doses of terabhivir, dexborneol, and their combinations increased the time APP / PS1 mice spent on the "novel arm." Specifically, administration of low-dose terabhivir combined with dexborneol combination 1 (T50 mg / kg + B2.5 mg / kg) and high-dose terabhivir combined with dexborneol combination 2 (T100 mg / kg + B2.5 mg / kg) significantly increased the time APP / PS1 mice spent on the "novel arm," with effects superior to terabhivir or dexborneol alone, demonstrating a synergistic effect. (Data are expressed as mean ± standard error; number of mice in each group n = 6; **P < 0.01 vs normal control group;) # P<0.05 ## P<0.01 vs APP / PS1+ solvent group; + P<0.05 ++ P<0.01 vs APP / PS1 + drug alone group).

[0282] The results show that the pharmaceutical composition containing terabhivir and dexborneol of the present invention can significantly improve the learning and memory abilities and cognitive functions of Alzheimer's disease mice, has a neuroprotective effect, and exhibits a synergistic effect. The pharmaceutical composition containing terabhivir and dexborneol can be used to prepare drugs for the prevention and treatment of Alzheimer's disease.

[0283] (3) Effects of telaprevir and camphor combination on spatial learning and memory ability and cognitive function in Alzheimer's disease model mice—Morris water maze test.

[0284] Following the requirements of the water maze experiment, the Morris water maze was divided into four quadrants. A platform with a diameter of 12 cm and a height of 35 cm (the target quadrant) was placed in one quadrant, submerged in water by 1-2 cm, and the water was dyed black with carbon ink. After five days of orientation and navigation experiments, the platform was removed, and a spatial exploration experiment was conducted. The time it took for the mice to reach the original platform location (the latency period) and the time spent in the target quadrant (the time spent in the quadrant where the original hidden platform was located) were recorded, and the percentage of time spent in the target quadrant relative to the total exploration time was analyzed. The animals' movement trajectories and behaviors were recorded using the Smart 3.0 small animal analysis software system, and the relevant data were analyzed.

[0285] Experimental results are as follows Figure 5 As shown in the figure, it can be concluded that compared with the normal control group mice, the time it took for the APP / PS1+ solvent group mice to find the original platform location (escape latency) was significantly increased (see Figure 1). Figure 5 A) The time spent in the target quadrant and the percentage of time spent in the target quadrant were significantly reduced (see A) Figure 5 B) Treatment of APP / PS1 mice with low and high doses of terabhivir, dexborneol, or their combinations significantly reduced the time to find the original platform (escape latency) and significantly increased the time spent in the target quadrant and the ratio of time spent in the target quadrant. Specifically, treatment with low-dose terabhivir combined with dexborneol combination 1 (T 50 mg / kg + B 2.5 mg / kg) and high-dose terabhivir combined with dexborneol combination 2 (T 100 mg / kg + B 2.5 mg / kg) significantly reduced the time to find the original platform (escape latency) and significantly increased the time spent in the target quadrant and the ratio of time spent in the target quadrant. These effects were superior to those of terabhivir or dexborneol alone, demonstrating a synergistic effect. (Data are expressed as mean ± standard error; number of mice in each group n = 6; **P < 0.01 vs. normal control group;) # P<0.05 ## P<0.01 vs APP / PS1+ solvent group; + P<0.05 ++ P<0.01 vs APP / PS1 + drug alone group).

[0286] Conclusion: The pharmaceutical composition containing telaprevir and dexborneol of the present invention can significantly improve the learning and memory abilities and cognitive functions of Alzheimer's disease mice, exhibiting neuroprotective effects and synergistic effects. The pharmaceutical composition containing telaprevir and dexborneol can be used to prepare drugs for the prevention and treatment of Alzheimer's disease.

[0287] Example 6

[0288] To verify the efficacy of a pharmaceutical composition containing telaprevir and camphor in combating amyotrophic lateral sclerosis (ALS).

[0289] Animal experiments: SOD1, one of the animal models of ALS, was used. G93A The (SOD1) mouse model was used to verify the anti-amyotrophic lateral sclerosis effect of a drug composition containing terabitvir and camphor.

[0290] Laboratory animals: ALS model SOD1 G93A The SOD1 mouse model was purchased from the company. It is known that SOD1 mice have an SOD1 gene mutation (SOD1). G93A This image perfectly reproduces the pathological condition of human ALS. Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease that selectively affects upper and lower motor neurons.

[0291] SOD1 G93A The transgenic ALS mouse model was purchased from an animal model company. All experimental animals were housed in an SPF-grade housing with a temperature of 22℃±2℃, a relative humidity of 45%±15%, free access to water, and following a 12-hour light / dark cycle.

[0292] Experimental grouping: Experimental animals were randomly divided into groups of 6-10. Based on the results of the preliminary experiment, the drug concentration and administration time were set. The drug groups were treated with different concentrations of terabitvir, camphor, and a combination of the two drugs.

[0293] Normal control group: SOD1 G93A (SOD1) transgenic background mice, such as Balb / c and C57BL / 6J mice + solvent.

[0294] SOD1 G93A +Solvent group (solvent group): SOD1 G93A Mice + vehicle.

[0295] SOD1 G93A +Telaprevir group: SOD1 G93A Mice treated with terabhivir.

[0296] SOD1 G93A +Borneol group: SOD1 G93A Mice treated with camphene.

[0297] SOD1 G93A +Terapitor group +Borneol group: SOD1 G93A Mice were treated with terabhivir and camphor.

[0298] The motor function and survival rate of mice were assessed using indicators such as the rotarod test, limb grip strength test, and survival time measurement.

[0299] (1) Rotarod experiment

[0300] The basic method is as follows: Three days before the test, the animals were allowed to acclimatize to the test environment for 30 minutes. Then, the mice were placed in a YLS-4C rotarod fatigue tester, starting at 5 r / min and increasing to 35 r / min within 180 seconds. The time the mouse spent on the rotarod when it fell was recorded; for mice that did not fall, 180 s / (35 r / min) was recorded. For the first experiment, the mice were allowed to acclimatize to the rotarod environment. Then, the experiment was repeated three times daily at the same time and under the same conditions, with a 30-minute rest interval between each test. The maximum value of the three results was taken.

[0301] (2) Grip strength test

[0302] The basic method is as follows: Mouse grip strength is used to directly assess the muscle strength of the mouse's limbs. The mouse is gently placed on the central platform of the gripping board, and its tail is gently pulled to encourage it to grasp the board. As the mouse grips the board, a further pull is applied to force it to release its grip. The maximum grip strength is recorded. This experiment is repeated three times, and the maximum value among the three results is taken as the evaluation value.

[0303] (3) Survival determination: For mice that underwent behavioral evaluation, the survival rate after administration of terabhivir and camphor was evaluated using Kaplan-Meier curves.

[0304] result:

[0305] Result 1. Behavioral tests in ALS model mice, SOD1 G93A Compared with the normal control group, the mice in the rat group showed significantly weakened grip strength in all four limbs and a significantly shorter time to fall off the rotisserie. Treatment with terabitvir or camphor monotherapy, or the combination of both drugs, delayed the onset of neurological symptoms in ALS model mice, with the combination of terabitvir and camphor being more effective than either drug alone.

[0306] Result 2. Compared with ALS model mice, treatment with terabitvir or camphor monotherapy or the combination of the two drugs improved the motor function of ALS model mice, significantly enhanced limb grip strength, and prolonged the time mice spent falling off the rotundus.

[0307] Result 3. Compared with the normal control group, the survival time of ALS model mice was significantly shortened. The survival time of mice treated with terabitvir or camphor alone or in combination was significantly longer than that of the ALS model group, and the survival time of mice in the combination treatment group was significantly better than that of mice treated with single drugs.

[0308] Conclusion: Treatment with terabhivir, camphor monotherapy, and the combination of the two drugs prolonged the survival time and improved the motor function of ALS mice. The combination therapy was more effective than the monotherapy, showing a synergistic effect. Terabhivir and camphor combination therapy can be used to treat amyotrophic lateral sclerosis.

[0309] Example 7

[0310] To verify the efficacy of a pharmaceutical composition containing telaprevir and camphor in combating amyotrophic lateral sclerosis (ALS).

[0311] 7.1 Animal experiments: One of the ALS animal models, B6SJL.SOD1, was used. G93A (abbreviated as SOD1) G93A A mouse model was used to investigate the anti-amyotrophic lateral sclerosis effect of the combination of terabitrin and dextromethorphan.

[0312] Experimental animals: ALS model was B6SJL.SOD1 G93A Mouse model, known SOD1 G93A Mice with SOD1 gene mutations reproduced the pathogenesis of human ALS very well. Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease that selectively affects upper and lower motor neurons.

[0313] SOD1 G93A The transgenic ALS mouse model was purchased from Cyagen (Suzhou) Biotechnology Co., Ltd. All experimental animals were housed in an SPF-grade housing with a temperature of 22℃±2℃, a relative humidity of 45%±15%, free access to water, and following a 12-hour light / dark cycle.

[0314] Experimental grouping and administration method: SOD1 G93AMice were randomly divided into groups of six at 60 days of age based on their motor function. Drug concentrations and administration times were determined according to preliminary experimental results. The drug groups were treated with different concentrations of telaprevir, dextroborneol, or a combination of both. Intraperitoneal injections were administered once daily starting at 71 days of age (11 weeks) until death. The normal control group and SOD1... G93A The mouse control group was given the same volume of solvent, which consisted of 10% DMSO + 30% PEG400 + 60% physiological saline.

[0315] Normal control group: C57BL / 6J mice + intraperitoneal injection of solvent;

[0316] SOD1 G93A +Solvent group (solvent group): SOD1 G93A Intraperitoneal injection of solvent in mice;

[0317] SOD1 G93A +Terrapivir low-dose group (+Terrapivir L): SOD1 G93A Mice were administered terabhivir (7.5 mg / kg) via intraperitoneal injection.

[0318] SOD1 G93A +Terrapivir high-dose group (+Terrapivir H): SOD1 G93A Mice were administered terabitril (23 mg / kg) via intraperitoneal injection.

[0319] SOD1 G93A +D-camphor group (+D-camphor): SOD1 G93A Mice were administered dextromethorphan (0.75 mg / kg) via intraperitoneal injection.

[0320] SOD1 G93A +Terrapivir low-dose combination with dextroborneol group 1 (+Combination 1): SOD1 G93A Mice were administered the combination of 1 and 2 (the composition of 1 satisfies the following conditions: the dose of terabitvir is 7.5 mg / kg and the dose of dexborneol is 0.75 mg / kg) via intraperitoneal injection.

[0321] SOD1 G93A +Terrapivir and dexborneol combination 2 groups (+Composition 2): SOD1 G93A Mice were administered the combination of 2 and 3 (the composition of 2 satisfies the following conditions: the dose of terabitvir is 23 mg / kg and the dose of dexborneol is 0.75 mg / kg) via intraperitoneal injection.

[0322] The motor function and survival rate of mice were assessed using indicators such as the rotarod test, limb grip strength test, and survival time measurement.

[0323] Statistical analysis: GraphPad Prism 9.0 software was used for statistical analysis. All data are expressed as mean ± standard error (±SEM). Independent samples Student's t-test was used to compare the means of two groups. One-way ANOVA was used to analyze differences among multiple groups. Tukey's test was used for post-hoc multiple comparisons. A p-value < 0.05 was considered statistically significant.

[0324] (1) Rotarod experiment

[0325] The basic method is as follows: Before the test (3 days), allow the animals to adapt to the test environment for 30 minutes. Then, place the mice in a YLS-4C rotundus fatigue tester, starting at 5 rpm and increasing to 30 rpm within 180 seconds. Specific steps: First, train the mice for 5 minutes with a rotation speed of 30 rpm, twice a day for three days. At the start of the test, conduct three consecutive tests (180 seconds each, with a 40-minute interval between each test). The experiment lasts 180 seconds, and the time it takes for any animal to fall off the platform is recorded as the latency period, or until the end of the experiment. Record the time the mouse spends on the rotundus each time. The longest time spent on the rotundus out of the three tests is taken as the mouse's motor function. If the time exceeds 180 seconds, the test is stopped, and 180 seconds is used as the calculation. SOD1 G93A Mice were tested for motor function using the aforementioned rotarod fatigue tester starting at 60 days of age, and then observed and tested once a week until the mice died.

[0326] (2) Grip strength test

[0327] The basic method is as follows: Mouse grip strength is used to directly assess the muscle strength of the mouse's limbs. The mouse is gently placed on the central platform of the gripping board, and its tail is gently pulled to encourage it to grasp the board. As the mouse grips the board, a further pull is applied to force it to release its grip. The maximum grip strength is recorded. This experiment is repeated three times, and the maximum value among the three results is taken as the evaluation value.

[0328] (3) Survival determination: For mice that underwent behavioral evaluation, the survival rate after administration of terabhivir and camphor was evaluated using Kaplan-Meier curves.

[0329] 7.2 Results:

[0330] Result 1. Rotating bar experiment and behavioral tests in ALS model mice: The experimental results are as follows. Figure 6 As shown, SOD1 G93A Compared with the normal control group, the mice in the mouse group showed significantly weakened muscle strength and a significantly shorter time to fall off the rotundus. (This is in contrast to SOD1.)G93A In mouse groups, treatment with low and high doses of terabhivir, dexborneol, and their combinations delayed SOD1 production. G93A Mice showed decreased muscle strength; among them, treatment with low-dose terabhivir combined with dexborneol combination 1 (T 7.5 mg / kg + B 0.75 mg / kg) and high-dose terabhivir combined with dexborneol combination 2 (T 23 mg / kg + B 0.75 mg / kg) resulted in decreased SOD1 levels at 84 days of age. G93A Mice remained on the rotarod for a significantly longer period (latency) than the ALS solvent control group, terabitvir, or dexborneol monotherapy group, which improved motor function and demonstrated a synergistic effect. (Data are expressed as mean ± standard error, n = 6, **P < 0.01 vs normal control group;) ## P<0.01 vs SOD1 G93A + Solvent group; + P<0.0<0.01 vs SOD1 G93A + Individual drug group).

[0331] Result 2. Compared with ALS model mice, treatment with terabitvir or camphor monotherapy or the combination of the two drugs improved the motor function of ALS model mice, significantly enhanced limb grip strength, and prolonged the time mice spent falling off the rotundus.

[0332] Result 3. Compared with the normal control group, the survival time of ALS model mice was significantly shortened. The survival time of mice treated with terabitvir or camphor alone or in combination was significantly longer than that of the ALS model group, and the survival time of mice in the combination treatment group was significantly better than that of mice treated with single drugs.

[0333] Conclusion: The pharmaceutical composition containing terabhivir and dexborneol of the present invention can significantly improve the motor function of mice, and the combined use is more effective than the single drug, showing a synergistic effect. The pharmaceutical composition of terabhivir and dexborneol can be used to prepare drugs for the prevention and treatment of amyotrophic lateral sclerosis (ALS).

[0334] Example 8

[0335] Verify the anti-Parkinson's effect of a pharmaceutical composition containing telaprevir and camphene.

[0336] Animal experiments: PD animal models were prepared using 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) to verify the anti-Parkinson's effect of a drug composition containing terabhivir and camphene.

[0337] Establishment of the MPTP animal model: A Parkinson's disease (PD) animal model was established using 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). Male C57BL / 6 mice, 10–12 weeks old and weighing 25–30 g, were given three days of pre-treatment acclimatization, and mice with poor motor coordination were removed. 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) was injected intraperitoneally at a dose of 30–40 mg / kg body weight for 5–7 consecutive days. Temporary symptoms such as trunk shaking, piloerection, tail hyperextension, reduced movement, and difficulty climbing poles appeared after the 5th–7th injection, indicating successful establishment of the Parkinson's disease (PD) mouse model.

[0338] The Parkinson's disease score is as follows: 0 points, similar to normal mice, with no symptoms; 1 point, with piloerection, arched back, and intermittent fine tremors, but still able to move freely; 2 points, with frequent swallowing, frequent tremors, hind limbs outstretched, tail tremors, and gradually restricted movement; 3 points, with drooling, persistent tremors, stiff limbs, and restricted movement; 4 points, death due to general paralysis.

[0339] Experimental grouping: Experimental animals were randomly divided into groups of 6-10. Based on preliminary experimental results, drug concentrations and administration times were set. The drug groups were treated with different concentrations of terabitvir, camphor, and a combination of both drugs, respectively.

[0340] Normal control group (Control group): C57BL / 6 mice.

[0341] MPTP model group: MPTP intraperitoneal injection for 5-7 days.

[0342] Telaprevir + MPTP model group: Telaprevir was administered after modeling.

[0343] Borneol + MPTP model group: Borneol was administered after modeling.

[0344] Telaprevir + Borneol + MPTP model group: Telaprevir and Borneol were administered during modeling.

[0345] Testing methods: The muscle strength and motor balance of the limbs of mice were tested using methods such as the pole climbing test and stiffness score.

[0346] (1) Pole test: The basic method is as follows: A rough wooden ball with a diameter of 9 mm is placed on top of a rough wooden stick with a circular cross-section and a length of 75 cm. The bottom of the stick is placed in a mouse cage. The mouse is placed on the ball and trained one day in advance to climb to the bottom of the stick with its head down. On the day of the experiment, the time it takes for the mouse to climb to the top of the stick is recorded as A, and the time it takes to climb to the bottom of the stick is recorded as B. The time it takes for the mouse to climb the entire stick is C, where C = AB. Each mouse is tested 3 to 5 times, and the average time of the 3 to 5 climbs is used as the statistical index.

[0347] (2) Rigidity scoring: The basic method is as follows: The mouse is held in a fixed position with its forelimbs placed on a 1cm wide wooden stick 4cm above the horizontal plane. A stopwatch is used to record the duration of this posture. Timing stops when both forelimbs are removed from the stick or the animal moves its head in an exploratory manner. The cutoff time is 300s. Each mouse is tested 3 times, with a 1-minute interval between each test. If the mouse maintains this posture for more than 30s, it is considered rigid. A score is given based on the time: 0s = 0 points, 1s-150s = 1 point, and 151s-300s = 2 points.

[0348] Experimental results:

[0349] Compared with the control group, the time required for mice to climb the pole in the MPTP model group was significantly increased, and the rigidity score was significantly elevated. Treatment with terabitvir or camphor alone or in combination significantly shortened the time for mice to climb the pole and reduced the rigidity score. The combination of terabitvir and camphor was more effective than either drug alone.

[0350] Conclusion: The combined use of terabhivir and camphene is more effective than the single drug, showing a synergistic effect. It can significantly improve motor function and rigidity symptoms in Parkinson's mice, reduce neuronal death, and has a neuroprotective effect. It can be used to prepare anti-Parkinson's drugs.

[0351] Example 9

[0352] Verify the anti-multiple sclerosis effect of a pharmaceutical composition containing telaprevir and camphor.

[0353] Animal experiments: The efficacy of a drug combination containing terapeptide and camphor against multiple sclerosis was verified using an experimental autoimmune encephalomyelitis (EAE) mouse model.

[0354] Experimental animals: 6-8 week old female C57BL / 6J mice, weighing 18-25 grams. All experimental animals were housed in an SPF-grade housing for one week at a temperature of 22℃±2℃, relative humidity of 45%±15%, with free access to water and following a 12-hour light / dark cycle. Then, modeling and drug administration were carried out according to the requirements of each experimental group.

[0355] Establishment of an experimental autoimmune encephalomyelitis (EAE) mouse model:

[0356] The basic experimental steps are as follows: First, prepare MOG. 35～55 The peptide was diluted with PBS to a concentration of 3 mg / ml (MOG); Mycobacterium tuberculosis was added to Freund's incomplete adjuvant (CFA) to prepare a concentration of 5 mg / ml Freund's complete adjuvant; MOG and CFA were mixed in a 1:1 ratio to prepare an antigen adjuvant emulsion. After anesthetizing mice, 0.2 ml / mouse was injected subcutaneously into the neck and back of the neck, divided into four injection points (randomly selected four points along the spine), 50 μl at each point. Subsequently, 0.1 ml (3 μg / ml, i.e., 300 ng) of pertussis toxin was injected intraperitoneally. 48 hours later, 0.1 ml of pertussis toxin was injected intraperitoneally again. After immunization with MOG, mice were routinely fed and their general condition was observed. The first day of immunization was recorded as Day 0. Then, at the same time every day, two experimenters scored the neurological dysfunction of each group of mice until 28 days after MOG immunization. The modified Kono 5-point scoring scale was used for scoring, and the specific scoring details are as follows:

[0357] 0 points (no clinical symptoms); 0.5 points (decreased tail tension, with the tip drooping); 1 point (tail dragging on the ground); 1.5 points (incomplete paralysis of one hind limb); 2 points (complete paralysis of one hind limb, able to roll over independently); 2.5 points (complete paralysis of one hind limb, accompanied by incomplete paralysis of the other hind limb); 3 points (complete paralysis of both hind limbs, unable to roll over independently, but can move on the ground after stimulation); 3.5 points (complete paralysis of both hind limbs, accompanied by paralysis of one forelimb); 4 points (paralysis of all four limbs or accompanied by urinary and fecal incontinence); 5 points (dying state or death).

[0358] The mice's food intake, water intake, activity, weight, and fur were recorded. A score of 0.5 to 5 was used to determine if a mouse was diseased. The time from the establishment of the model to the onset of the disease was recorded as the incubation period, and the time from the onset of the disease to the peak of the disease (no increase in the neurological dysfunction score for 3 consecutive days) was recorded as the progression period. Diseased mice were sacrificed at the peak of the disease, while unaffected mice and normal control mice were observed until 28 days after the establishment of the model and then sacrificed.

[0359] Experimental grouping: Experimental animals were randomly divided into groups of 6-10. Based on preliminary experimental results, drug concentrations and administration times were set. The drug groups were treated with different concentrations of terabitvir, camphor, and a combination of both drugs, respectively.

[0360] Normal control group (Control group): C57BL / 6J mice without model + solvent.

[0361] EAE Model Group (EAE Group): Model Group + Solvent.

[0362] Telaprevir+EAE model group: Telaprevir was administered after modeling.

[0363] Borneol + EAE model group: Borneol was administered after modeling.

[0364] Telaprevir + Borneol + EAE model group: Telaprevir and Borneol were administered after modeling.

[0365] Detection methods: The modified Kono 5-point scale was used to detect the neurological function of mice and to detect changes in body weight.

[0366] result:

[0367] Effects of telaprevir, camphor monotherapy, and combination therapy on neurological function and body weight in mice.

[0368] Compared with the control group, the EAE model group showed clinical symptoms such as decreased tail tension and increased neurological function scores 7-11 days after MOG immunization. Treatment with terabitvir or camphor monotherapy or the combination of the two drugs could reduce the neurological function scores of mice and improve their neurological function. The combination of terabitvir and camphor was more effective than the monotherapy.

[0369] Conclusion: The combined use of telaprevir and camphene is more effective than the single drug, showing a synergistic effect. It can significantly improve the neurological function of EAE mice, reduce neuronal death, and has a neuroprotective effect. It can be used to prepare drugs against multiple sclerosis for the treatment of multiple sclerosis.

[0370] The above embodiments show that the drug composition containing telaprevir and camphene can be used to treat cerebral ischemia / reperfusion injury (including ischemic stroke), myocardial ischemia / reperfusion injury (myocardial infarction), Alzheimer's disease, amyotrophic lateral sclerosis (ALS), Parkinson's disease, and multiple sclerosis. The combination therapy is more effective than monotherapy, showing a synergistic effect, which helps to ensure efficacy and reduce drug dosage, thereby improving the safety of clinical drug use while ensuring clinical treatment effect.

[0371] Reference 1: Zou C, Mifflin L, Hu Z, Zhang T, Shan B, Wang H, Xing X, Zhu H, Adiconis X, Levin JZ, Li F, Liu CF, Liu JS, Yuan J. Reduction of mNAT1 / hNAT2Contributes to Cerebral Endothelial Necroptosis and AβAccumulation in Alzheimer's Disease.Cell Rep.2020;33(10):108447.

[0372] However, this invention is not limited to the above-mentioned diseases; the drug is also applicable to the treatment of other diseases with similar pathogenesis.

[0373] The above embodiments should be understood as being used only to illustrate the present invention more clearly, and not to limit the scope of the present invention. After reading the present invention, any modifications of the present invention in various equivalent forms by those skilled in the art fall within the scope defined by the appended claims.

Claims

1. A pharmaceutical composition, characterized in that, It contains terabhivir and camphene.

2. The pharmaceutical composition according to claim 1, characterized in that, The mass ratio of terabhivir to camphene is 1:0.001-1000, preferably 1:0.01-100, more preferably 1:0.05-25, even more preferably 1:0.1-10, even more preferably 1:0.1-5, even more preferably 1:0.15-1, and even more preferably 1:0.2-0.

8.

3. The pharmaceutical composition according to claim 1, characterized in that, The mass ratio of terapeptide to camphor is 3-30:0.25-10, preferably 5-30:0.5-8, more preferably 7.5-30:0.75-6, even more preferably 10-30:0.8-2, and even more preferably 12-30: 0.9-1.5。 4. The pharmaceutical composition according to claim 1, characterized in that, The camphor includes one or more of dextrocamphor, levoncamphor, and racemic camphor; preferably, the pharmaceutical composition further includes a pharmaceutically acceptable carrier.

5. The use of the pharmaceutical composition according to any one of claims 1-4 in the preparation of a cell protection drug.

6. The application according to claim 5, characterized in that, The cell-protective drug is a drug used to prevent, protect, and / or treat cell damage caused by pathological conditions and / or degeneration processes; the pathological conditions and / or degeneration processes include situations that may lead to cell death, preferably including pathological apoptosis and / or pathological necrosis and / or necrotizing apoptosis and / or pyroptosis and / or ferroptosis and / or disulfide-dependent cell death and / or autophagy (anti-apoptotic drugs and / or anti-necrotizing drugs and / or anti-pyroptosis drugs and / or antiferroptosis drugs and / or anti-dependent cell death drugs and / or disulfide-dependent cell death drugs and / or anti-autophagy drugs) and / or diseases or conditions; preferably, the diseases or conditions include, but are not limited to: nervous system diseases, cardiovascular system diseases, hemorrhage. Contraindicated in thrombotic diseases, diffuse connective tissue diseases, organ-specific or systemic inflammation or autoimmune diseases, autoimmune diseases, bone diseases, joint and cartilage diseases, ischemic diseases or attacks of limbs, ophthalmic diseases, skin diseases, kidney diseases, blood and vascular diseases, lung diseases, gastrointestinal diseases, liver diseases, metabolic diseases, muscle diseases, pancreatic diseases, severe poisoning by chemicals, infectious agents, toxins or drugs, age-related diseases, dental diseases, auditory conduction pathway diseases, mitochondrial-related diseases, and / or trauma and / or exposure to biological and / or chemical and / or physical and / or medical and / or surgical procedures, such as accidental infarction and hemorrhage, and / or medical and / or surgical procedures, such as cell, tissue or organ transplantation.

7. The application according to claim 5 or 6, characterized in that, The cell-protective drugs mentioned above refer to drugs that have the effect of preventing, inhibiting or treating damage, degeneration or dysfunction of tissues, organs and cells caused by hypoxia / reoxygenation; Alternatively, the cell-protective drug refers to a drug that has the effect of preventing, inhibiting or treating damage, degeneration or dysfunction of tissues, organs and cells caused by necroptosis; Preferably, the neurological diseases include stroke, transient ischemic attack, focal ischemia, intracranial hemorrhage, prenatal hypoxia, adult or childhood hypoxia, neurodegenerative diseases, muscle diseases, trigeminal neuralgia, glossopharyngeal neuralgia, Bell's palsy, progressive bulbar palsy, primary lateral sclerosis (PLS), pseudobulbar palsy, invertebral disc syndrome, cervical spondylosis, plexus disorder, thoracic outlet destruction syndrome, porphyria, peripheral neuropathy, multiple system atrophy, corticobasal degeneration, progressive supranuclear palsy, Lewy body dementia, demyelinating diseases, frontotemporal dementia, Gullman-Barré syndrome, multiple sclerosis, Kreutzfeldt-Jacob disease, progressive peroneal muscular atrophy, prion disease, and lethal family members. The following are included in the list of neurodegenerative diseases: insomnia, G. Schereschat syndrome, bovine spongiform encephalopathy, epilepsy, hereditary ataxia, Friedreich ataxia, spinocerebellar ataxia, hereditary spastic paraplegia, dystonia, multiple system atrophy, lysosomal storage diseases, Niemann-Pick disease, Gaucher disease, AIDS dementia syndrome, neurological damage caused by exposure to toxic compounds consisting of industrial solvents, heavy metals, drugs, and chemotherapeutic agents, and neurological damage caused by mechanical, physical, or chemical trauma; preferably, stroke includes one or more of ischemic and hemorrhagic strokes; preferably, the neurodegenerative disease includes Alzheimer's disease. Muscle diseases include one or more of the following: Parkinson's disease, amyotrophic lateral sclerosis (ALS), infantile spinal muscular atrophy, Huntington's disease, and Parkinson's plus syndrome; preferably, the muscle diseases include one or more of the following: muscular atrophy, Duchenne's muscular dystrophy, ankylosing spondylitis, myopathy and myasthenia gravis, myasthenia gravis, progressive muscular atrophy, spinal muscular atrophy, and hereditary muscular atrophy; Preferably, the cardiovascular diseases include one or more of the following: myocardial ischemia and / or vascular ischemia, ischemic heart disease, angina pectoris, unstable angina pectoris, refractory angina pectoris, myocardial infarction, myocardial ischemia / reperfusion injury, hypoxia, low oxygen, chronic or acute heart failure, systolic heart failure and diastolic heart failure, left ventricular dysfunction, post-myocardial infarction left ventricular dysfunction, hypertensive heart disease, rheumatic heart disease, cardiomyopathy, myocardial hypertrophy, hypertrophic cardiomyopathy, myocarditis, valvular heart disease, arrhythmia, paroxysmal tachycardia, atrial fibrillation, ventricular fibrillation, arteriosclerosis, atherosclerosis, peripheral vascular disease, aneurysm, peripheral vascular occlusive disease (preferably cerebral occlusion, pulmonary occlusion or intestinal occlusion), chronic venous insufficiency or varicose veins, hypertension, systemic hypertension, pulmonary hypertension, portal hypertension, and cardiovascular toxic side effects caused by drug (preferably anticancer drugs) treatment; Preferably, the stroke includes ischemic stroke; Preferably, the diffuse connective tissue disease includes one or more of the following: rheumatoid arthritis, juvenile idiopathic arthritis, lupus erythematosus, systemic lupus erythematosus, scleroderma, idiopathic inflammatory myopathy, polymyositis, dermatomyositis, vasculitis, necrotizing vasculitis, polyarteritis nodosa, granulomatous vasculitis, giant cell arteritis, Sjögren's syndrome, systemic scleroderma, allergic cutaneous vasculitis, and Behcet's disease; Preferably, the organ-specific inflammation, systemic inflammation, or autoimmune disease includes one or more of the following: chronic inflammatory bowel disease, bronchial asthma, chronic obstructive pulmonary disease, eosinophilic sinusitis, and systemic lupus erythematosus. Preferably, the ophthalmic diseases or conditions include one or more of the following: diabetic retinopathy, glaucoma, retinal degeneration, retinitis pigmentosa, corneal retinal dystrophy, optic neuropathy and optic neuritis, optic drusen, ptosis, chronic progressive extraocular muscle paralysis, macular degeneration, retinal tear or retinal rupture, retinal ischemia, retinal ischemia / reperfusion injury, retinal detachment, trauma-related acute retinopathy, inflammatory degeneration, postoperative complications, drug-induced retinopathy or cataract, wet or dry AMD-related photoreceptor degeneration; Preferably, the skin disease includes one or more of the following: dermatitis, psoriasis, scarring, aging or altered healing process, eczema, and collagen diseases; Preferably, the trauma and / or exposure to factors of biological and / or chemical and / or physical and / or medical and / or surgical origins include severe poisoning caused by infectious agents, toxins, chemicals, or drugs. More preferably, the symptoms of severe poisoning include one or more of sepsis, septic shock and its consequences, or iatrogenic diseases.

8. The application according to claim 5 or 6, characterized in that, The cell-protective drug is one or more drugs used to prevent and / or protect and / or treat cell death in transplanted organs and / or organ donors and / or organ recipients, prevent acute transplant rejection of organs and / or increase long-term survival rates, limit primary organ dysfunction and / or limit the delay in the recovery of transplanted organ function and / or improve the functional recovery of transplanted organs. Preferably, the cell-protective drug is a drug for preventing and / or protecting and / or treating pathological conditions or degenerative processes associated with ischemia / reperfusion symptoms; more preferably, the pathological conditions or degenerative processes associated with ischemia / reperfusion symptoms include one or more of actual cold ischemia, warm ischemia, actual reperfusion, and ischemia / reperfusion phenomena. Preferably, the cell-protective drug is a drug used to prevent and / or protect and / or treat organs, tissues or cells against ischemia / reperfusion injury. Optionally, the ischemia / reperfusion injury is caused by cold or warm ischemia and / or reperfusion and / or ischemia / reperfusion. Further, the ischemia / reperfusion injury includes one or more of the following: cerebral ischemia / reperfusion injury, myocardial ischemia / reperfusion injury, hepatic ischemia / reperfusion injury, renal ischemia / reperfusion injury, pulmonary ischemia / reperfusion injury, intestinal ischemia / reperfusion injury, and limb ischemia / reperfusion injury. Preferably, the cell-protective drug is a drug for the prevention and / or protection and / or treatment of one or more of the following: heart failure caused by infarction, neurological sequelae caused by stroke or trauma, tissue damage affecting liver, intestine, heart, lung or kidney transplantation to grafts or the consequences of surgical procedures. Alternatively, the cell-protective drug is a drug for preventing and / or protecting and / or treating one or more of nerve cells (a drug for protecting the brain), heart cells (a drug for protecting the heart), liver cells (a drug for protecting the liver), kidney cells (a drug for protecting the kidneys), intestinal cells, or lung cells. Preferably, the cell-protective drug is a drug for protecting nerve cells, vascular endothelial cells, brain endothelial cells, and / or cardiomyocytes.

9. Use of the pharmaceutical composition according to any one of claims 1-4 in the preparation of organ preservation solutions.

10. An organ preservation solution, characterized in that, It comprises the pharmaceutical composition as described in any one of claims 1-4; Preferably, the concentration of telaprevir in the organ preservation solution is from 0.01 mg / L to 1000 mg / L, more preferably from 0.1 mg / L to 100 mg / L, and even more preferably from 1 mg / L to 10 mg / L; and the concentration of camphene is from 0.1 mg / L to 150 mg / L, more preferably from 1 mg / L to 50 mg / L, and even more preferably from 1 mg / L to 10 mg / L.

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