WO2022048197A1 - UTILISATION D'α-ASARONE DANS LA PRÉPARATION DE MÉDICAMENT DE PRÉVENTION OU DE TRAITEMENT D'UN ACCIDENT VASCULAIRE CÉRÉBRAL ISCHÉMIQUE - Google Patents

UTILISATION D'α-ASARONE DANS LA PRÉPARATION DE MÉDICAMENT DE PRÉVENTION OU DE TRAITEMENT D'UN ACCIDENT VASCULAIRE CÉRÉBRAL ISCHÉMIQUE Download PDF

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WO2022048197A1
WO2022048197A1 PCT/CN2021/095672 CN2021095672W WO2022048197A1 WO 2022048197 A1 WO2022048197 A1 WO 2022048197A1 CN 2021095672 W CN2021095672 W CN 2021095672W WO 2022048197 A1 WO2022048197 A1 WO 2022048197A1
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ischemic stroke
emulsion
asarone
group
oil
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PCT/CN2021/095672
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Chinese (zh)
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毛声俊
张坤
冯欣倩
刘琪
张检
张迪
母珂蔓
高小凤
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四川大学
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/075—Ethers or acetals
    • A61K31/085—Ethers or acetals having an ether linkage to aromatic ring nuclear carbon
    • A61K31/09—Ethers or acetals having an ether linkage to aromatic ring nuclear carbon having two or more such linkages
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis

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  • the present invention relates to the application of ⁇ -asarone in preparing medicine for preventing or treating ischemic stroke.
  • Stroke is now the second leading cause of adult death and the leading cause of adult disability. In China, although the mortality rate of stroke is generally decreasing, the incidence rate is increasing day by day. Currently, there are about 2.4 million new cases every year. Stroke can be clinically divided into hemorrhagic stroke and ischemic stroke. Ischemic stroke is mainly caused by atherosclerosis leading to thrombus formation or thrombus shedding and blocking cerebral blood vessels. When an ischemic stroke occurs, the amount of blood circulating in the brain is reduced or absent, leading to neuronal damage and death.
  • Cerebral ischemia and reperfusion can cause a variety of pathological cascades in the brain, including energy metabolism disorders, oxidative stress, inflammation, cytokine damage, glutamate toxicity, Ca 2+ overload, excess nitric oxide (NO) ) synthesis, apoptosis, etc.
  • the cascade first manifests as excitotoxicity, in the central area of the infarct, due to ischemia leading to ATP depletion, inactivation of the sodium pump, and excessive release of excitatory amino acids (glutamate) from the presynaptic membrane, resulting in neuronal damage.
  • the release of excitatory neurotransmitters and the depolarization of neuronal cell membranes are the key links in cell damage and death caused by acute ischemic stroke.
  • the main goal of stroke treatment is to rescue hypoperfused, nonfunctional but viable tissue around the ischemic penumbra or infarct core.
  • the clinical treatment strategy for ischemic stroke is mainly cerebral blood flow reconstruction, including intravenous thrombolysis and intra-arterial thrombolysis, as well as thrombectomy under the guidance of angiography and fluoroscopy.
  • Recombinant tissue plasminogen activator (rtPA) is mainly used in thrombolytic therapy, but rtPA thrombolysis, as a main and effective treatment method, has a narrow time window for treatment (symptoms). Within 3 to 4.5 hours after appearance), less than 10% of patients are suitable.
  • neuroprotective agents can not only reduce nerve cell death, but also prolong the time window of thrombolytic therapy. Therefore, the development of effective and safe neuroprotective agents is of great significance.
  • GABA Gamma-aminobutyric acid
  • prophylactic antiepileptic drugs are generally not recommended (Chinese Medical Association Neurology Branch, Chinese Medical Association Neurology Branch Cerebrovascular Disease Group, China Acute Epilepsy Guidelines for Diagnosis and Treatment of Hemorrhagic Stroke 2018 [J]. Chinese Journal of Neurology, 2018, 51(9): 666-682.), the reason is that antiepileptic drugs have serious side effects, and preventive administration of antiepileptic drugs may damage ischemic Neurological function in stroke patients.
  • the present invention provides the application of ⁇ -asarone in the prevention or treatment of ischemic stroke.
  • the first aspect of the present invention provides the compound shown in formula I (trans-2,4,5-trimethoxy-1-propenylbenzene, also known as ⁇ -asarone, ⁇ -asarone) in the preparation of The application of drugs in the prevention or treatment of ischemic stroke;
  • the present invention unexpectedly finds that the compound represented by formula I ( ⁇ -asarone) can significantly improve the neurological symptoms of ischemic stroke model rats and reduce the volume of cerebral infarction, and it has a precise effect on ischemic stroke animal models. Therapeutic effect, and no obvious toxic side effects.
  • ⁇ -asarone, ⁇ -asarone, positive drug butylphthalide emulsion injection and edaravone injection are used to treat middle cerebral artery occlusion (Middle cerebral artery occlusion, MCAO) cerebral ischemia-regeneration Perfusion model rats, it was found that ⁇ -asarone can significantly increase the blood flow ratio of the infarcted hemisphere and the contralateral hemisphere, reduce the volume of cerebral infarction, significantly improve the neurological function score, improve the survival rate, and prolong the survival rate of the model rats. Expect. The experimental results show that ⁇ -asarone has a significantly better therapeutic effect on ischemic stroke than ⁇ -asarone.
  • the present invention also unexpectedly found that ⁇ -asarone is significantly more effective than butylphthalide emulsion injection in reducing the incidence of secondary epilepsy caused by ischemic stroke; in terms of anti-neuroinflammation, the effect is remarkable Better than edaravone injection.
  • the medicament is also used to prevent or treat secondary epilepsy caused by ischemic stroke.
  • the medicament is used to treat ischemic stroke and prevent secondary epilepsy due to ischemic stroke.
  • the drug can reduce the expression of LC3 protein in brain tissue.
  • the medicament can also reduce the expression of GFAP and Iba-1 proteins in brain tissue, inhibit neuroinflammation, reduce autophagy, apoptosis and necrosis of neurons, thereby exerting an anti-ischemic stroke effect.
  • the compound represented by formula I is the only active ingredient in the medicine.
  • the medicament may contain pharmaceutical excipients.
  • the total weight ratio of the compound represented by the formula I to the pharmaceutical excipients is 1:20-200. More preferably, the compound represented by the formula I is the only active ingredient in the medicine, and the total weight ratio of the compound to the pharmaceutical excipients is 1:20-200.
  • the daily effective dose of the compound represented by formula I in the medicine when used in the treatment of ischemic stroke model rats, can be 10 mg-40 mg/kg body weight.
  • the object of prevention or treatment of the drug is a human or an animal.
  • the daily dose of the compound represented by the formula I in the medicament can range from 0.5 mg to 5.0 mg/kg body weight, preferably 1.0 mg ⁇ 4.0 mg/kg body weight. If it is administered 2-3 times a day, the dosage range of each administration can be 0.3 mg-2.0 mg/kg body weight, preferably 0.5 mg-1.0 mg/kg body weight. The above doses can be obtained according to the dose conversion relationship between different species of animals.
  • the drug administration object can be human or animal. Described medicine can be made into the dosage form that is suitable for human and/or animal use, for example any dosage form that is compatible with different routes of administration, as long as the dosage form can make the compound shown in formula I enter the brain and reach effective therapeutic concentration.
  • the route of administration of the drug is injection, oral administration, subcutaneous implantation, inhalation, transdermal, mucosal and the like.
  • the dosage form is an injection or an oral formulation.
  • the dosage form is an intravenous injection or an oral preparation.
  • the compound shown in the formula I is used for clinical treatment of ischemic stroke patients, it can be diluted with an appropriate amount of sodium chloride injection before use, and the mode of administration is intravenous drip, which is conducive to better maintenance. its effective blood concentration.
  • the medicament is an emulsion (eg, emulsion injection, oral emulsion).
  • the emulsion may contain the compound represented by formula I, a pharmaceutically acceptable oil, a pharmaceutically acceptable emulsifier and water.
  • the pharmaceutically acceptable oil can be composed of at least one of soybean oil, medium chain oil, olive oil and fish oil.
  • the pharmaceutically acceptable emulsifier may be composed of at least one of egg yolk lecithin, soybean lecithin, Pluronic F-68, and polyethylene glycol stearic acid-15 (Solutol HS15).
  • the water can be water for injection or purified water.
  • the emulsion can also contain at least one of oleic acid and sodium oleate.
  • oleic acid is dissolved in the oil phase
  • sodium oleate is dissolved in the water phase
  • the mixture of the two can be dissolved in the oil and water phases respectively.
  • the emulsion can also contain glycerin.
  • the emulsion may also contain antioxidants.
  • the antioxidant can be sodium bisulfite, vitamin E, pyrogallate and the like.
  • the emulsion when administered orally, may further contain at least one of other suitable additives such as preservatives and flavoring agents.
  • suitable additives such as preservatives and flavoring agents.
  • the preservatives can be conventional preservatives in the art, such as benzoic acid, sodium benzoate, sorbic acid, potassium sorbate, ethylparaben, propyl ester and butyl ester and the like.
  • the flavoring agents can be conventional flavoring agents in the art, such as sweeteners, aromatic agents, mucilage agents or effervescent agents.
  • the sweeteners can be simple syrup, stevioside, aspartame, etc.; the aromatics can be fruit flavors, such as apple flavor, strawberry flavor, etc.; the mucilage agents can be gelatin, methyl cellulose gum
  • the effervescent agent can be a mixture of citric acid, tartaric acid and sodium bicarbonate.
  • the emulsion may contain 0.5% to 5% of the compound represented by formula I, 5% to 30% of pharmaceutically acceptable oil, 0.6% to 1.8% of emulsifier, and 0% of glycerin in terms of weight percentage. ⁇ 2.5%, and the balance water (eg purified water or water for injection).
  • concentration of the compound of formula I in the emulsion can vary within a certain range, and the range of concentration variation depends on the dosage, the dosage volume and the solubility of the compound of formula I in the oil phase.
  • the emulsion may contain ⁇ -asarone 0.5%-5%, pharmaceutically acceptable oil 5%-20%, emulsifier 0.6%-1.8%, glycerin 0% by weight percentage ⁇ 2.5%, and the balance water (eg purified water or water for injection).
  • the emulsion is an injection emulsion.
  • the total weight ratio of the compound represented by formula I to pharmaceutical excipients is 1:20-200.
  • the preparation method of described emulsion may comprise the following steps: the compound shown in formula I, pharmaceutically acceptable oil, pharmaceutically acceptable emulsifier and water are mixed by high-speed shearing to obtain colostrum; Homogenize to obtain an emulsion.
  • the preparation method of the emulsion may include the following steps:
  • Step 1 Under the protection of nitrogen or inert gas, the compound represented by formula I is dissolved in a pharmaceutically acceptable oil at 60-80 ° C to obtain an oil phase, and then the emulsifier and glycerol are dissolved or dispersed at 60-80 ° C. or, under the protection of nitrogen or inert gas, the compound represented by formula I and the emulsifier are dissolved or dispersed in a pharmaceutically acceptable oil at 60-80 ° C to obtain an oil phase, and then the glycerol Dissolve in water at 60 ⁇ 80°C to obtain water phase;
  • Step 2 mixing the above-mentioned oil phase and water phase through high-speed shearing, so that the oil phase is dispersed in the water phase to obtain colostrum;
  • Step 3 The colostrum is homogenized by high pressure (for example, the number of times of homogenization can be 1 to 3 times), so that the average particle size of the milk droplets is not greater than 0.5 ⁇ m, filtered, and poured under the protection of nitrogen or inert gas. Packed in glass ampoules, infusion bottles, vials, soft bags and other medicinal containers; according to the needs of the route of administration, use rotary autoclaving or add preservatives without sterilization to obtain an emulsion.
  • high pressure for example, the number of times of homogenization can be 1 to 3 times
  • the shear rate of the high-speed shearing can be a conventional shear rate used in the preparation of emulsions in the art, for example, it can be 2000-4000 r.min-1, and for example, 2500-3500 r.min-1.
  • the shearing time of the high-speed shearing can be the conventional shearing time used in the preparation of emulsions in the art, for example, it can be 3-10 minutes, and for example, it can be 5-8 minutes.
  • the homogenization pressure of the high-pressure homogenization can be the conventional homogenization pressure used in the field to prepare the emulsion, for example, it can be 500-1500 bar, and for example, it can be 500-1000 bar.
  • the cycle times of the high-pressure homogenization can be the conventional cycle times used in the preparation of emulsions in the art, for example, it can be 1 to 6 times.
  • the present invention also provides a pharmaceutical composition for preventing or treating ischemic stroke, wherein the pharmaceutical composition contains the compound represented by formula I and pharmaceutical excipients.
  • the pharmaceutical composition is also used to prevent or treat secondary epilepsy caused by ischemic stroke.
  • the pharmaceutical composition is used to treat ischemic stroke and prevent secondary epilepsy caused by ischemic stroke.
  • the compound of formula I is the only active ingredient in the pharmaceutical composition.
  • the pharmaceutical composition is an emulsion.
  • the present invention also provides a method for treating or preventing ischemic stroke in a subject, comprising: administering to the subject a therapeutically or prophylactically effective amount of the compound represented by formula I.
  • the method is for treating or preventing ischemic stroke and treating or preventing epilepsy secondary to ischemic stroke in a subject.
  • the present invention also provides a method for treating or preventing ischemic stroke in a subject and treating or preventing secondary epilepsy caused by ischemic stroke, comprising: administering to the subject a therapeutically or prophylactically effective amount The compound of formula I.
  • trans-2,4,5-trimethoxy-1-propenylbenzene alias: ⁇ -asarone, ⁇ -asarone
  • alpha-asarone can also be used to prevent and treat epilepsy associated with ischemic stroke.
  • ⁇ -asarone is combined with pharmaceutically acceptable excipients to prepare a dosage form suitable for human or animal use, and the route of administration can be injection, oral administration, subcutaneous implantation, inhalation, transdermal , mucosa, etc., the dosage form can be any dosage form suitable for different administration routes, provided that the dosage form can enable ⁇ -asarone to enter the brain to achieve an effective therapeutic concentration.
  • alpha-asarone is the only active ingredient of the drug for the treatment of ischemic stroke.
  • the dosage forms of the medicine for treating ischemic stroke are injections and oral preparations.
  • ⁇ -asarone can be used as the only active ingredient in the medicine, and the total weight ratio of ⁇ -asarone to pharmaceutically acceptable auxiliary materials can be 1:20-200.
  • the daily effective dose of the drug for treating ischemic stroke can be 1.0-10.0 mg/kg body weight, for example, 2-10 mg/kg body weight.
  • the preferred dosage forms are emulsions.
  • the emulsion may comprise an ischemic stroke-treating effective amount of alpha-asarone and a pharmaceutically acceptable oil as an oil phase; a pharmaceutically acceptable emulsifier as an emulsifier; and the remainder as an aqueous phase.
  • the medicinal oil can be composed of at least one of soybean oil, medium chain oil, olive oil, and fish oil.
  • the pharmaceutically acceptable emulsifier may be composed of at least one of egg yolk lecithin, soybean lecithin, Pluronic F-68, and polyethylene glycol stearic acid-15 (Solutol HS15).
  • oleic acid, sodium oleate or a mixture of the two in any ratio can also be added according to the needs of emulsifying performance.
  • oleic acid is dissolved in the oil phase
  • sodium oleate is dissolved in the water phase
  • the mixture of the two is dissolved in the oil and water phases respectively.
  • the water phase can be water for injection or purified water.
  • the aqueous phase may also contain glycerol.
  • the total weight ratio of ⁇ -asarone and pharmaceutically acceptable excipients may be 1:20-200.
  • the weight ratio of each component in the emulsion can be ⁇ -asarone 0.5% to 5%, oil phase 5% to 20%, emulsifier 0.6% to 1.8%, glycerol 0% to 2.5%, purified water or water for injection. to 100%.
  • the ⁇ -asarone in an effective amount for treating ischemic stroke means that the ⁇ -asarone contained in the emulsion can effectively treat the above diseases.
  • the effective amount varies depending on the subject of administration (eg, humans or animals).
  • the administration volume of the emulsion should be appropriate to the administration subject.
  • the concentration of alpha-asarone in the emulsion can vary within a certain range. The concentration range varies depending on the dose administered, the volume administered and the solubility of alpha-asarone in the oil phase. For example, the concentration can range from 0.5% to 5.0%, ie, 5 mg/mL to 50 mg/mL.
  • the emulsion may also contain antioxidants, and the antioxidants may include sodium bisulfite, vitamin E, pyrogallate and the like.
  • antioxidants may include sodium bisulfite, vitamin E, pyrogallate and the like.
  • other suitable additives such as preservatives and flavoring agents may also be used.
  • the dosage form of the medicine for treating ischemic stroke is injection or oral preparation.
  • the pharmaceutical dosage form is an injection
  • the preferred dosage form is an emulsion.
  • the total weight ratio of ⁇ -asarone to pharmaceutically acceptable excipients can be 1:20 ⁇ 200; the weight ratio of each component in the emulsion can be 0.5% ⁇ 200% of ⁇ -asarone 5%, oil phase 5%-20%, emulsifier 0.6%-1.8%, glycerol 0%-2.5%, purified water or water for injection added to 100%.
  • ⁇ -asarone when used as a drug for treating ischemic stroke in humans, its effective therapeutic dose range may be 1.0 mg-10.0 mg/kg body weight, preferably 1.5 mg-4.0 mg/kg.
  • the preparation method includes the following steps:
  • the colostrum is homogenized by high pressure for 3 to 6 times, so that the average particle size of the milk droplets is not greater than 0.5 ⁇ m, and filtered to obtain a drug-loaded emulsion containing ⁇ -asarone; under nitrogen protection, it is potted and sealed
  • suitable medicinal containers such as glass ampoules, infusion bottles, etc.; according to the needs of the route of administration, it can be obtained by rotary autoclaving in a suitable way.
  • the preparation method comprises the following steps:
  • the colostrum is homogenized under high pressure so that the average particle size of the milk droplets is not greater than 0.5 ⁇ m, and filtered to obtain a drug-loaded emulsion containing ⁇ -asarone, which is filled with suitable medicines such as glass ampoules and infusion bottles.
  • suitable medicines such as glass ampoules and infusion bottles.
  • a suitable method to sterilize by rotary autoclaving In a container; according to the needs of the route of administration, use a suitable method to sterilize by rotary autoclaving.
  • the effective dose for the treatment of ischemic stroke in rats may be 10 mg to 40 mg/kg body weight.
  • the effective therapeutic dose can range from 1.0 mg to 10.0 mg/kg body weight. , preferably 1.5 mg to 4.0 mg/kg body weight.
  • the present invention can be used as a medicament for the treatment of ischemic stroke in humans, as well as for the treatment of epilepsy caused by ischemic stroke in humans. It can be administered by injection or oral route.
  • the terms “pharmaceutically acceptable” and “pharmaceutically acceptable” refer to those compounds, materials, compositions and/or dosage forms that are within the scope of sound medical judgment. It is suitable for use in contact with human and animal tissues without undue toxicity, irritation, allergic reactions or other problems or complications, commensurate with a reasonable benefit/risk ratio.
  • pharmaceutically acceptable excipients means “pharmaceutical excipients”, which refer to the excipients and additives used in the production of pharmaceutical products and in the formulation of prescriptions, which are included in pharmaceutical preparations in addition to active ingredients. of all substances. See the Pharmacopoeia of the People's Republic of China (2020 Edition) Volume Four, or the Handbook of Pharmaceutical Excipients (Raymond C Rowe, 2009 Sixth Edition).
  • treatment refers to therapeutic therapy.
  • treatment refers to: (1) ameliorating one or more biological manifestations of the disease or disorder, (2) interfering with (a) one or more points in the biological cascade leading to or causing the disorder or (b) ) one or more biological manifestations of the disorder, (3) amelioration of one or more symptoms, effects or side effects associated with the disorder, or one or more symptoms, effects or side effects associated with the disorder or its treatment, or (4) slowing the progression of the disorder or one or more biological manifestations of the disorder.
  • the term “therapeutically effective amount” refers to an amount of a compound that, when administered to a subject, is sufficient to treat the disease or disorder described herein.
  • a “therapeutically effective amount” will vary depending on the compound, the condition and its severity, and the age of the patient to be treated, but can be adjusted as needed by those skilled in the art. The effective amount varies depending on the subject of administration (eg, humans or animals).
  • prevention refers to a reduced risk of acquiring or developing a disease or disorder.
  • prophylactically effective amount refers to an amount sufficient to prevent a disease or disorder, or an amount sufficient to prevent one or more symptoms associated with a disease or disorder, or to prevent recurrence of a disease or disorder.
  • the term “subject” refers to any animal, preferably a mammal, and most preferably a human, to which the compound is to be or has been administered according to embodiments of the present invention.
  • the term “mammal” includes any mammal. Examples of mammals include, but are not limited to, cows, horses, sheep, pigs, cats, dogs, mice, rats, rabbits, guinea pigs, monkeys, humans, etc., with humans being the most preferred.
  • the reaction temperature is room temperature, which is generally 20-35°C.
  • secondary epilepsy due to ischemic stroke in the present invention (also referred to herein as epilepsy associated with ischemic stroke, epilepsy caused by ischemic stroke) refers to The patient has no history of epilepsy, and secondary epileptic seizures caused by ischemic stroke (excluding lesions not related to ischemic stroke).
  • the reagents and raw materials used in the present invention are all commercially available.
  • the present invention discovers for the first time that ⁇ -asarone has the effect of treating/preventing ischemic cerebral apoplexy, and provides a safe and effective medicine for the treatment and prevention of ischemic cerebral apoplexy.
  • the results of pharmacodynamic mechanism studies show that ⁇ -asarone can significantly increase the middle cerebral artery by reducing the expression of GFAP, Iba-1 and LC3 proteins in brain tissue, inhibiting neuroinflammation and neuronal autophagy, apoptosis and necrosis.
  • the blood flow ratio of the infarcted side hemisphere and the contralateral hemisphere of the occlusive ischemia-reperfusion (MCAO) model rats reduces the volume of cerebral infarction; significantly improves the neurological function score, improves the survival rate, and prolongs the survival period; significantly Reduce the incidence of secondary epilepsy due to ischemic stroke.
  • MCAO occlusive ischemia-reperfusion
  • the present invention unexpectedly found that the therapeutic effect of ⁇ -asarone on ischemic stroke is significantly better than that of ⁇ -asarone; In terms of incidence, the effect was significantly better than that of the butylphthalide administration group; in terms of anti-neuroinflammation, the effect was significantly better than that of edaravone injection. Accordingly, ⁇ -asarone is expected to become a promising drug for the treatment of ischemic stroke.
  • Figure 1 The effect of ⁇ -asarone on apoptosis of brain tissue in rats with cerebral ischemia-reperfusion.
  • A TUNEL staining pictures of rat brain tissue after 24 hours of cerebral ischemia and reperfusion
  • B Percentage of TUNEL staining positive cells in brain tissue of rats in each group
  • P, M, L, H, E represent sham-operated group, model group, ⁇ -Asarone low-dose group, ⁇ -Asarone high-dose group, Edaravone injection group
  • TUNEL means terminal deoxynucleotidyl transferase-mediated dUTP end-labeling technology
  • DAPI is 4′, 6- Diamidino-2-phenylindole
  • Merge indicates the merge of TUNEL staining pictures and DAPI staining pictures
  • arrows indicate TUNEL staining positive cells; compared with model group: ***P ⁇ 0.001, ns means no significant difference.
  • Figure 2 The effect of ⁇ -asarone on brain histomorphology in rats with cerebral ischemia-reperfusion.
  • Figure 3 Results of Nissl staining and quantitative analysis of Nissl bodies in brain tissue of cerebral ischemia-reperfusion rats.
  • Figure 4 The effect of ⁇ -asarone on the expression of GFAP in brain tissue of cerebral ischemia-reperfusion rats.
  • GFAP is glial fibrillary acidic protein, a specific marker of astrocytes; arrows indicate GFAP; compared with model group: *P ⁇ 0.05, **P ⁇ 0.01; compared with sham operation group: ### P ⁇ 0.001; ns means no significant difference.
  • Figure 5 The effect of ⁇ -asarone on the expression of Iba-1 in brain tissue of cerebral ischemia-reperfusion rats.
  • Iba-1 is an ionic calcium adaptor protein, which is a marker of microglia; arrows indicate Iba-1; compared with the model group: *P ⁇ 0.05; compared with the sham operation group: # P ⁇ 0.05; ns means no significance difference.
  • Figure 6 The effect of ⁇ -asarone on the expression of LC3 in brain tissue of cerebral ischemia-reperfusion rats.
  • LC3 is an autophagy-related protein and a marker of autophagosome formation; arrows indicate LC3; compared with model group: *P ⁇ 0.05, **P ⁇ 0.01; compared with sham operation group: ## P ⁇ 0.01; ns Indicates no significant difference.
  • the above oil phase is added to the water phase, sheared at high speed for 5 to 15 minutes, and water is added to 1000 mL to prepare colostrum.
  • water is added to 1000 mL to prepare colostrum.
  • a high-pressure homogenizer for 3 to 6 times, so that the average particle size of the homogenized milk droplets is not greater than 0.5 ⁇ m, filter through the filter membrane, and fill the filtrate into 5mL-20mL glass ampoules under the protection of nitrogen.
  • rotary autoclaving is performed at 121° C. for 8-12 minutes to obtain ⁇ -asarone injection emulsion, wherein the concentration of ⁇ -asarone is 0.5-50 mg/mL.
  • the above oil phase is added to the water phase, sheared at high speed for 5 to 15 minutes, and water is added to 1000 mL to prepare colostrum.
  • a high-pressure homogenizer for 3 to 6 times, so that the average particle size of the homogenized milk droplets is not greater than 0.5 ⁇ m, filter through a filter membrane, and fill the filtrate into 5mL or 10mL glass ampoules under nitrogen protection.
  • the above oil phase is added to the water phase, sheared at high speed for 5 to 15 minutes, and water is added to 1000 mL to prepare colostrum.
  • a high-pressure homogenizer for 3 to 6 times, so that the average particle size of the homogenized milk droplets is not greater than 0.5 ⁇ m, filter through a filter membrane, and fill the filtrate into 5mL or 10mL glass ampoules under nitrogen protection.
  • the preparation method is the same as that of Preparation Example 1.
  • the oil phase can add a pharmaceutically acceptable amount of antioxidants such as vitamin E, pyrogallate, etc., and the oil phase can also add a pharmaceutically acceptable amount of preservatives, such as paraben Ethyl ester, etc.;
  • the water phase can add pharmaceutically acceptable amount of flavoring agents, such as fruit juice syrup with aromatic flavor;
  • the water phase can also add pharmaceutically acceptable amount of preservatives, such as benzoic acid, sodium benzoate, etc.
  • Colostrum is prepared by the same method, and then the colostrum is homogenized by a high-pressure homogenizer for 3 to 6 times, so that the average particle size of the homogenized milk droplets is not greater than 10 ⁇ m, filtered through a filter membrane, and the filtrate is filled under nitrogen protection.
  • suitable medicinal packaging circulating steam is sterilized at 100 DEG C for 15-45 minutes, or 121 DEG C for 8-12 minutes to obtain ⁇ -asarone oral emulsion.
  • the above oil phase is added to the water phase, sheared at high speed for 5 to 15 minutes, and water is added to 1000 mL to prepare colostrum.
  • a high-pressure homogenizer for 1 to 3 times, so that the average particle size of the homogenized milk droplets is not greater than 0.5 ⁇ m, filter through the filter membrane, and fill the filtrate in 2mL, 5mL, and 10mL under the protection of nitrogen.
  • rotary autoclave sterilization at 121° C. for 8 to 12 minutes to obtain ⁇ -asarone milky injection, wherein the content of ⁇ -asarone is 1 mg/mL to 20 mg/mL.
  • the above-mentioned oil phase was added to the water phase, and sheared at high speed (2800r.min-1) for 10 minutes to disperse the oil phase in the water phase, and water was added to 1000L to prepare colostrum.
  • a high-pressure homogenizer and the homogenization pressure is 1000bar, so that the average particle size of the milk droplets after homogenization is not greater than 0.5 ⁇ m, filter membrane, and fill the filtrate in 2mL under nitrogen protection.
  • the above-mentioned oil phase was added to the water phase, and sheared at high speed (rotation speed: 2500 r ⁇ min -1 ) for 10 minutes to disperse the oil phase in the water phase, and water was added to 1000 L to prepare colostrum.
  • the homogenization pressure is 800bar, so that the average particle size of the milk droplets after homogenization is not greater than 0.5 ⁇ m, filter membrane, and fill the filtrate in 10mL under nitrogen protection.
  • Oral vial circulating steam sterilization at 100°C ⁇ 30min, or rotary autoclave sterilization at 121°C ⁇ 8min to obtain ⁇ -asarone oral emulsion, wherein the content of ⁇ -asarone is 10 mg/mL.
  • mice were randomly divided into sham operation group (P group), model group (M group, given the same volume of blank emulsion as the milky injection high-dose group), and ⁇ -asarone cerebral intravenous administration group (with the same preparation).
  • Example 7 method operation prepared into milky injection, the concentration is 10mg/mL, the dosage is 10mg/kg, B group), ⁇ -asarone brain milky injection low-dose group (made by Preparation Example 7, 10mg/kg, L group), high-dose group of ⁇ -asarone brain emulsion injection (made by Preparation Example 7, 20mg/kg, H group), ⁇ -asarone brain emulsion oral administration group (implemented by preparation Example 8 made, 40mg/kg, O group), butylphthalide group (because the concentration of commercially available butylphthalide sodium chloride injection is as low as 0.25mg/ml is inconvenient to administer, so operate with preparation example 7 method, Prepared into butylphthalide emulsion injection, the concentration is 10mg/mL, the dosage is 10mg/kg, C group), 12 in each group.
  • ⁇ -asarone brain milky injection low-dose group made by Preparation Example 7, 10mg/kg,
  • the MCAO model was made by the modified suture method: the rats were fasted for 12 hours before operation, and the rats were anesthetized by intraperitoneal injection of 10% chloral hydrate (350 mg/kg). Supine fixation, midline neck incision, blunt dissection of the muscle and fascia along the inner border of the sternocleidomastoid muscle, separation of the right common carotid artery (CCA), external carotid artery (ECA) and internal carotid artery (ICA). CCA proximal end, ICA and ECA disposal lines are reserved. The CCA and ECA were ligated, and the ICA was temporarily clamped with an arterial clip.
  • CCA common carotid artery
  • ECA external carotid artery
  • ICA internal carotid artery
  • the rats were scored after anesthesia and awake. Those with a score of 1 to 3 were included in the group:
  • the rats were anesthetized by intraperitoneal injection of 10% chloral hydrate (350 mg/kg), and the brains were harvested after cardiac perfusion with normal saline, and then the brains were placed in a -20°C refrigerator for 20-30 min to facilitate slicing.
  • the cerebellum, olfactory bulb, and lower brainstem were removed. Coronal section, 2mm thick, the first incision is at the midpoint of the line connecting the anterior pole of the brain and the optic chiasm; the second incision is in the optic chiasm; the third incision is in the funnel peduncle; the fourth incision is in the funnel peduncle and the caudal pole of the posterior lobe between.
  • the brain slices were placed in 2% TTC dye solution pre-warmed at 37°C for 30 minutes, protected from light, and the container was slightly shaken every 5 minutes to fully stain the brain slices.
  • the brain slices were taken out, fixed with 4% paraformaldehyde solution for 24 hours and photographed.
  • the infarct area and right brain area were measured by Image J image analysis system, and the relative infarct volume of the brain was calculated.
  • Relative infarct volume total area of cerebral infarction/total area of right brain. The results are shown in Table 2.
  • 1.6 ⁇ -Asarone increases the relative blood flow of the infarcted hemibrain in rats with cerebral ischemia-reperfusion
  • the rats were anesthetized with 10% chloral hydrate (350 mg/kg) intraperitoneally, placed in the prone position, the skull was exposed, and the bilateral cerebral hemisphere blood flow was monitored using a laser speckle contrast imaging system.
  • the MoorFLPI image analysis system was used to open the cerebral blood flow images, and 6mm ⁇ 4mm areas were selected in the same part of the left and right hemispheres to measure the blood flow of the left and right hemispheres.
  • the relative blood flow value of the infarct side hemi-cerebral blood flow the blood flow value of the infarct side hemisphere/the contralateral hemi-cerebral blood flow value. The results are shown in Table 2.
  • the oral administration group of ⁇ -asarone cerebral emulsion (group O), Butylphthalide emulsion injection group (group C) can significantly improve the neurological symptoms of model rats, reduce the size of cerebral infarction, and increase the relative value of hemi-cerebral blood flow on the infarcted side (P ⁇ 0.01); ⁇ -asarone cerebral vein
  • group B the results of this study unexpectedly found that ⁇ -asarone has a precise therapeutic effect on cerebral ischemia-reperfusion injury.
  • the experimental materials, grouping and modeling methods are the same as those in Example 1.
  • the rats are ischemia for 2 hours. Immediately after reperfusion, they are administered according to the grouped dosing schedule. After that, the administration is continued for 14 days, once a day.
  • the model group and the sham operation The group was given the same volume of normal saline as the high-dose ⁇ -asarone brain milk injection group. The survival of the rats was observed and recorded, and the results are shown in Table 3.
  • Seizure grade refers to the Racine standard, which divides seizure behavior into 6 grades according to the degree of seizures: grade 0 is no response or seizure stops; grade I is rhythmic mouth or facial twitching; grade II is nodding or tail flicking; grade III is Single-limb twitching; grade IV twitching or rigidity of multiple limbs; grade V generalized tonic-clonic seizures. Classes I, II, and III are clonic seizures, and classes IV and V are tonic seizures.
  • Tunel incubation solution (batch number: 11684795910, Roche, Switzerland);
  • DAPI dye solution (batch number: DA0001, Beijing Regen Biology Co., Ltd.);
  • Glycerol gelatin solution (batch number: PH1425, Tianjin Ruijinte Chemicals Co., Ltd.);
  • Anti-fluorescence decay mountant (batch number: AR1109, Wuhan Boster Biological Engineering Co., Ltd.);
  • Goat serum blocking solution (batch number: ZLI-9021, Beijing Zhongshan Jinqiao Biological Co., Ltd.);
  • DAB color development kit (batch number: K135925C, Beijing Zhongshan Golden Bridge Biological Co., Ltd.);
  • APES 3-aminopropyltriethoxysilane, batch number: 16A06A01, Boster Biotechnology Co., Ltd.
  • GFAP primary antibody (batch number: bs-0199R, Beijing Boaosen Biotechnology Co., Ltd.);
  • Iba-1 primary antibody (batch number: GB12105, Beijing Boaosen Biotechnology Co., Ltd.);
  • LC3 primary antibody (batch number: ab58610, Abcam);
  • Goat anti-rabbit working solution (batch number: SP-9001, Beijing Zhongshan Jinqiao Biological Co., Ltd.);
  • FITC-labeled goat anti-rabbit IgG (batch number: GB22303, Servicebio);
  • the modeling method of SD rats is the same as item "1.1" in Example 1. After modeling, they were randomly divided into a sham operation group (P), a model group (M), and a low-dose group of ⁇ -asarone brain milky injection (prepared by Prepared in Example 7, 10 mg/kg, L group), high-dose ⁇ -asarone-brain emulsion injection group (produced by Preparation Example 7, 20 mg/kg, H group), Edaravone injection group ( Commercially available edaravone injection specification 5mL: 10mg, 3mg/kg, E group), 5 rats in each group, 2 hours after cerebral ischemia, immediately after reperfusion according to the grouped dosing schedule, the corresponding drugs or with ⁇ -Blank emulsion of the same volume in the high-dose group of Asarum-brain emulsion injection.
  • anesthesia was performed with 10% chloral hydrate (350 mg/kg, i.p.). Open the chest and abdomen to fully expose the heart, liver, and lungs. Cut the superior and inferior vena cava, insert the perfusion needle into the left ventricle, first perfuse the heart with about 20 mL of 0.9% normal saline until the lungs are white, and then perfuse with about 20 mL of 4% paraformaldehyde until the body and After the organs were hardened, the brain tissue was separated by decapitation and fixed in 4% paraformaldehyde for 72 hours.
  • Figure 2 shows that after 24 hours of cerebral ischemia and reperfusion, the cortex of the model group (M group) had obvious neuronal degeneration and necrosis, some neurons were vacuolated, cytoplasmic vacuolated, and pyknosis was deep. Compared with the M group, the pathological damage of the brain tissue in the E, L and H groups was significantly reduced.
  • the low-dose ⁇ -asarone brain emulsion group (L group) had no significant effect on the expression of LC3 in the brain of cerebral ischemia-reperfusion rats; the ⁇ -asarone high-dose group ( Group H) can significantly reduce the content of LC3 in the brain of cerebral ischemia-reperfusion rats, and there is no significant difference compared with the positive drug.
  • mice were randomly divided into five groups, blank control group (Blank group, given a blank emulsion equivalent to the administration volume of ⁇ -asarone high-dose group, 10 animals), cyclophosphamide group (CTX group) , 40mg/kg, 10 rats), ⁇ -asarone brain milky injection low-dose group (made by Preparation Example 7, 100 mg/kg/day, ASA-L group, 10 rats), ⁇ -asarone brain milky injection Injection medium-dose group (made by Preparation Example 7, 150 mg/kg/day, ASA-M group, 10), ⁇ -Asarone-brain emulsion injection high-dose group (made by Preparation Example 7, 200 mg /kg/day, ASA-H group, 10 animals).
  • blank control group Breast group, given a blank emulsion equivalent to the administration volume of ⁇ -asarone high-dose group, 10 animals
  • CTX group cyclophosphamide group
  • CTX group cyclophos
  • mice All drugs were injected through the tail vein. Except for the positive control drug, cyclophosphamide (CTX), which was injected only 24 hours before the sampling, the other groups were continuously administered through the tail vein for 4 days, and were sacrificed 24 hours after the last administration.
  • CX cyclophosphamide
  • mice after separating both sides of the femur, the femoral bone marrow cells were washed out with PBS, passed through a 300-mesh nylon mesh to make a single cell suspension, centrifuged at 1650 rpm for 5 min, and resuspended in PBS to adjust the cell concentration to 5 ⁇ 10 6 cells/mL.
  • PIPES-PI solution 10 mL of PIPES solution (3.5 mg/mL) + 0.5 mg PI + 0.01 mL of Triton X-100 (0.1% concentration), gently pipetting and mixing. After staining at 4°C for 30 min in the dark, flow cytometry was used for detection, and the results are shown in Table 5.
  • PCE is polychromatic erythrocytes
  • MNPCE is polychromatic erythrocytes with micronuclei
  • fMNPCE is polychromatic erythrocytes with micronuclei.
  • the ratio of polychromatic erythrocytes to polychromatic erythrocytes reflects the micronucleus rate of mouse bone marrow cells. The higher the value, the stronger the genotoxicity.
  • the micronucleus rate of the positive control drug cyclophosphamide (CTX) administration group Compared with the blank emulsion group, it was significantly improved (P ⁇ 0.01); compared with the CTX group, each dose group of ⁇ -asarone injection had statistical differences (ASA-L: P ⁇ 0.01; ASA-M: P ⁇ 0.01). 0.05; ASA-H: P ⁇ 0.05).

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Abstract

La présente invention concerne l'utilisation d'un composé, représenté par la formule I, dans la préparation d'un médicament de prévention ou de traitement d'un accident vasculaire cérébral ischémique. Il a été découvert pour la première fois dans la présente invention que l'α-asarone a les effets de prévention et de traitement d'un accident vasculaire cérébral ischémique, ce qui fournit un médicament sûr et efficace pour la prévention et le traitement d'un accident vasculaire cérébral ischémique.
PCT/CN2021/095672 2020-09-04 2021-05-25 UTILISATION D'α-ASARONE DANS LA PRÉPARATION DE MÉDICAMENT DE PRÉVENTION OU DE TRAITEMENT D'UN ACCIDENT VASCULAIRE CÉRÉBRAL ISCHÉMIQUE WO2022048197A1 (fr)

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KR101355483B1 (ko) * 2011-06-15 2014-02-05 건국대학교 산학협력단 수검초 추출물 또는 이로부터 분리된 알파-아사론을 유효성분으로 함유하는 퇴행성 신경질환 예방 및 치료용 조성물

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CN110772606A (zh) * 2019-12-11 2020-02-11 孟胜喜 防治缺血性卒中的中药组合物及其制备方法

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* Cited by examiner, † Cited by third party
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Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
CAI, ZHOUQUAN ET AL.: "Preparation and Quality Evaluation of Intravenous α-Asarone Emulsion", WEST CHINA JOURNAL OF PHARMACEUTICAL SCIENCES, vol. 22, no. 6, 31 December 2007 (2007-12-31), pages 611 - 613, XP055906378 *
KIM, YOUNG-OCK;KWON, YONG-SOO;LEE, SEONG-EUN;LEE, SANG-WON;LEE, HEE-JAE: "Effects of alpha-Asarone against Global Cerebral Ischemia in Rats", NATURAL PRODUCT SCIENCES, vol. 15, no. 4, 31 December 2009 (2009-12-31), KR , pages 198 - 202, XP053006065, ISSN: 1226-3907 *
LEE HONG JU, AHN SUNG MIN, PAK MALK EUN, JUNG DA HEE, LEE SEO-YEON, SHIN HWA KYOUNG, CHOI BYUNG TAE: "Positive effects of α-asarone on transplanted neural progenitor cells in a murine model of ischemic stroke", PHYTOMEDICINE, vol. 51, 1 December 2018 (2018-12-01), AMSTERDAM, NL , pages 151 - 161, XP055906371, ISSN: 0944-7113, DOI: 10.1016/j.phymed.2018.09.230 *
XU, FEI-FEI ET AL.: "Research Progress on the Neuropharmacological Effects and Mechanisms about the Main Components α- and β-Asarone in Volatile Oils of Acorus Tatarrinowii Schott", NATURAL PRODUCT RESEARCH AND DEVELOPMENT, vol. 32, no. 11, 29 July 2020 (2020-07-29), pages 1969 - 1979, XP055906386 *

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