Application of colchicine in preparation of medicine for preventing and treating cerebral apoplexy

Abstract

The invention belongs to the field of biological medicines, and particularly discloses application of colchicine serving as an effective component for preventing and / or treating ischemic cerebrovascular diseases in preparation of medicines for preventing and / or treating ischemic cerebrovascular diseases. Tests prove that colchicine can effectively improve neurological impairment, reduce the cerebral infarction area, relieve the edema degree of brain tissues and reduce the level of inflammatory factors. The symptoms of the cerebral apoplexy are prevented and relieved from multiple angles, and the progress of the disease course is blocked; the medicine is safe, reliable and free of obvious toxic and side effects.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine, and in particular, relates to the use of colchicine in preparing a drug for preventing and / or treating cerebral stroke. Background Art

[0002] Cerebrovascular disease (CVD) includes transient ischemic attack (TIA) and stroke. Among them, stroke has surpassed cardiovascular disease and malignant tumors to become the leading cause of death and disability among Chinese residents, causing huge economic and social burdens, including acute ischemic stroke (AIS) and intracerebral hemorrhage (ICH). AIS, also known as acute cerebral infarction, is a group of clinical syndromes caused by various reasons that result in obstruction of brain tissue blood supply, ischemic hypoxic necrosis, and then neurological dysfunction. Acute ischemic stroke is the most common type of stroke, accounting for about 60%-80% of all strokes. However, the treatment of acute ischemic stroke is very limited. At present, the clinically proven effective acute treatment options are intravenous alteplase (tPA) thrombolysis or intravascular thrombectomy to achieve vascular recanalization in the ischemic area. However, due to the narrow treatment time window and the increased risk of complications such as cerebral edema and hemorrhagic transformation, intravascular thrombectomy requires a professional team with complex techniques, and clinically only 15% of patients can benefit from it.

[0003] According to domestic and foreign experience, stroke is preventable and controllable. Active and effective intervention in the risk factors of stroke can significantly reduce the incidence of stroke and reduce the burden of stroke. The risk factors of stroke are divided into two types: interveneable and non-interventional. Non-interventional factors mainly include: age, gender, race, genetic factors, etc.; interveneable factors include: hypertension, abnormal glucose metabolism, dyslipidemia, heart disease, asymptomatic carotid atherosclerosis and lifestyle [Guidelines for the Prevention and Treatment of Stroke in China (2021 Edition) http: / / www.nhc.gov.cn / yzygj / s3593 / 202108 / 50c4071a86df4bfd9666e9ac2aaac605.shtml]. Secondary prevention refers to early detection, early diagnosis, and early treatment of stroke in the formation and development process. In the early stage of ischemic stroke, appropriate secondary prevention strategies are extremely important. According to statistics, by adjusting the modifiable risk factors: adjusting diet, exercise, combined use of antithrombotic, lipid-lowering, and antihypertensive drugs, and controlling blood sugar, the control and treatment of cardiovascular diseases and atherosclerosis can reduce 80% of recurrent ischemic events [Xu Jie, Zhang Ying, Tan Sheng, Gao Qingchun. Analysis of the actual effect of secondary prevention drugs in patients with ischemic stroke and transient ischemic attack [J]. Chinese Journal of Modern Drug Application, 2022, 16(21): 1-6. DOI: 10.14164 / j.cnki.cn11-5581 / r.2022.21.001.].

[0004] Colchicine is the first-line drug for the treatment of acute gout, prevention of gout and familial Mediterranean fever. It has a strong anti-inflammatory effect because of its mechanism of action of inhibiting microtubule polymerization, changing the adhesion, mobility and cytokine production of leukocytes. It is also commonly used in clinical practice for other diseases [Tian Bingfeng, Yu Hualin, Ma Yiliu. Some progress in the clinical application of colchicine [J]. Journal of Kunming Medical College, 2009, 30(S1):107-111.], including pseudolupus erythematosus, pericarditis, Behcet's disease, cirrhosis, and neutrophilic dermatosis; it also has anti-tumor and anti-fibrosis effects. Low-dose colchicine can be used clinically for secondary prevention of cardiovascular diseases [Zheng Gang. The status and clinical significance of colchicine in the prevention and treatment of cardiovascular diseases [J]. Chinese Journal of Geriatric Cardiovascular and Cerebrovascular Diseases, 2022, 24(02): 216-218.], and is also a new target for secondary prevention of patients with ischemic cerebrovascular disease [Wu Jing, Zhou Shanshan, Chou Xiaohua, et al. Pharmaceutical practice of colchicine in the treatment of acute ischemic stroke [J]. Chinese Science and Technology Journal Database (Full Text Edition) Medicine and Health, 2022(5): 0005-0007.].

[0005] At present, the chemical drugs used to treat stroke are mainly thrombolytic drugs, anticoagulants and neuroprotectants. Among them, thrombolytic drugs are used to reduce blood clots and reduce the degree of vascular occlusion. Anticoagulants are mainly used to improve microcirculation and restore blood flow. Neuroprotectants are mainly used to maintain nerves and delay nerve damage. Traditional Chinese medicine is also widely used in the treatment of stroke. Single herbs and compound drugs mainly have the main effects of promoting blood circulation and removing blood stasis, such as Angelica sinensis, Panax notoginseng, Chuanxiong, etc.; compound drugs mainly include oral drugs such as Compound Danshen Tablets, Buyang Huanwu Decoction, Guanxin Shutong Capsules, Yiqi Qiangxin Capsules, Bushen Yiqi Capsules, and Naoxintong Capsules, as well as injection preparations such as Danhong Injection and Compound Danshen Injection. However, these drug compositions often have a single effect, complex composition, or poor safety, and lack strict and systematic pharmacodynamic experiments that meet the requirements of modern pharmacology, making it difficult to carry out strict quality control and standardized production.

[0006] However, the clinical application of colchicine lacks theoretical basis and specific mechanism of action, and in view of this, the present invention is proposed. Summary of the invention

[0007] The technical problem to be solved by the present invention is to overcome the deficiencies of the prior art, provide an application of colchicine in the preparation of a medicament for preventing and / or treating stroke, expand the application scope of colchicine, and provide experimental and theoretical basis for the clinical prevention and / or treatment of stroke-related diseases.

[0008] In order to solve the above technical problems, the basic concept of the technical solution adopted by the present invention is: The first object of the present invention is to provide a method for preparing a drug for preventing and / or treating a patient suffering from ischemic cerebrovascular disease by using colchicine as an effective ingredient for preventing and / or treating ischemic cerebrovascular disease.

[0009] In a further embodiment, the ischemic cerebrovascular disease in this technical solution is stroke, and preferably, the prevention is secondary prevention of stroke.

[0010] In a further solution, the stroke described in the technical solution highly expresses the following markers: IL-1 factor, IL-6 factor, NLRP3, TNF-α or IL-8 factor, inflammatory factors: A1AC (α1-antitrypsin), BPI (bactericidal / permeability-promoting protein), C5a (complement component 5a) and interleukin 1β (IL-1β). Preferably, the markers include: tumor necrosis factor-α (TNF-α), interleukin 6 (IL-6) and interleukin 1β (IL-1β).

[0011] In a further embodiment, the colchicine described in this technical solution is used as the only active ingredient acting on ischemic cerebrovascular disease.

[0012] The second object of the present invention is to provide a colchicine as an effective ingredient for preventing and / or treating ischemic cerebrovascular disease, and its use in the preparation of a drug for treating patients suffering from ischemic cerebrovascular disease, wherein the patients also suffer from: coronary heart disease, atrial fibrillation, hypertension, diabetes, obesity, chronic kidney disease and other diseases.

[0013] The third object of the present invention is to provide a pharmaceutical composition for preventing and / or treating ischemic cerebrovascular diseases, wherein the active ingredient for preventing and / or treating ischemic cerebrovascular diseases in the pharmaceutical composition includes colchicine.

[0014] In a further embodiment, the pharmaceutical composition described in this technical solution uses colchicine as the only active ingredient for preventing and / or treating ischemic cerebrovascular disease.

[0015] In a further embodiment, the pharmaceutical composition for preventing and / or treating ischemic cerebrovascular disease described in this technical solution further comprises a pharmaceutically acceptable carrier.

[0016] In a further embodiment, the pharmaceutical composition for preventing and / or treating ischemic cerebrovascular disease is an injectable drug or an oral drug.

[0017] In a further embodiment, the pharmaceutical composition for preventing and / or treating ischemic cerebrovascular disease is in the form of tablets, capsules, granules, suspensions, emulsions, solutions, syrups or injections.

[0018] After adopting the above technical solution, the present invention has the following beneficial effects compared with the prior art: The experimental data recorded in the present invention prove that colchicine can effectively improve neurological deficits, reduce the area of ​​cerebral infarction, reduce the degree of brain tissue edema, and reduce the level of inflammatory factors. It can prevent and alleviate the symptoms of stroke from multiple angles and block the progression of the disease. It is safe and reliable, and has no obvious toxic and side effects.

[0019] The specific implementation modes of the present invention are further described in detail below in conjunction with the accompanying drawings. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] The accompanying drawings are part of the present invention and are used to provide a further understanding of the present invention. The exemplary embodiments of the present invention and their descriptions are used to explain the present invention, but do not constitute an improper limitation of the present invention. Obviously, the drawings described below are only some embodiments. For ordinary technicians in this field, other drawings can be obtained based on these drawings without creative work. In the accompanying drawings: Figure 1Shown are the TTC-stained sections of the rat brain in each drug group; Figure A is the sham operation group, Figure B is the model group, Figure C is the nimodipine group, Figure D is the low-dose colchicine tablet group, and Figure E is the high-dose colchicine tablet group. Example

[0021] Effects of colchicine on cerebral ischemia-reperfusion model in rats 1. Experimental Materials 1.1 Test products and reagents Colchicine tablets were produced by Kunming Pharmaceutical Group Co., Ltd.; nimodipine tablets were produced by Bayer Healthcare Co., Ltd.; rat tumor necrosis factor (TNF-α), rat interleukin 6 (IL-6), and rat interleukin 1β (IL-1β) test kits were purchased from Beijing Solebow Technology Co., Ltd.; and sutures were purchased from Beijing Xilong Technology Co., Ltd.

[0022] 1.2 Experimental animals Male SD rats, 250-300 g, were obtained from Zhejiang Weitonglihua Experimental Animal Co., Ltd., with production license: SCXK(Zhejiang)2019-0001, issued by the Zhejiang Provincial Department of Science and Technology. Laboratory Animal Use License: SYXK(Yunnan)K2019-0001, issued by Kunming Science and Technology Bureau. The experimental animal room temperature was 20-25°C (daily temperature difference ≤3°C), humidity was 40-70%, lighting was 12h:12h light-dark alternation, illumination was 150-300lx, and noise was ≤60Db.

[0023] 2. Experimental Methods 2.1 Grouped Dosing 100 male SD rats were randomly divided into 5 groups after adaptive feeding for 1 week, with 20 rats in each group, namely sham operation group, model group, positive drug nimodipine group (10 mg / kg), low-dose colchicine (0.1 mg / kg) and high-dose colchicine group (0.2 mg / kg). The drugs in the nimodipine group and colchicine group were crushed and suspended with sodium carboxymethyl cellulose (CMC-Na) solution to prepare suspensions. They were shaken well before administration and given oral administration of the corresponding drugs every day. The sham operation group and model group were given the same amount of sodium carboxymethyl cellulose solution. The drugs were administered for 5 consecutive days, and the model was established on the 6th day. The drug was administered again after 1 hour of reperfusion.

[0024] 2.2 Preparation of rats with cerebral ischemia-reperfusion model The rats were fasted for 12 hours before the operation and were allowed to drink water freely. The rats were anesthetized by intraperitoneal injection of 10% chloral hydrate (0.3 ml / 100 g) and fixed on the operating table in supine position. The operation was performed according to the following steps: skin preparation and disinfection; midline incision in the neck to expose the common carotid artery, blunt separation of the internal carotid artery, temporary clamping of the common carotid artery and the internal carotid artery with an artery clamp, separation of the external carotid artery, ligation of the distal end of the external carotid artery, a small cut was made in the external carotid artery about 2-3 mm from the bifurcation of the common carotid artery, the thread plug was slowly and gently advanced through the incision of the external carotid artery toward the direction of the internal carotid artery entering the skull, a thin line was tied to the internal carotid artery and the thread plug was fixed so that it could be pushed without slipping out of the blood vessel, the external carotid vessel was shortened along the ligation at the distal end of the external carotid, the external carotid vessel was straightened to form a line with the internal carotid vessel, the artery clamp at the internal carotid artery was removed, and the thread plug was continued to be advanced, with the bifurcation of the common carotid artery as a mark, and slight resistance was encountered when advancing about 18 mm, that is, reaching the thinner middle cerebral artery and stopping, so that the local cerebral blood flow was fully blocked, the thin line on the external carotid artery was tightened, the artery clamp on the common artery was removed, the wound was sutured, and a thread plug about 3 cm long was left outside the body, and the surgical area was disinfected with iodine. After 1 hour of ischemia, the occluding wire was removed to achieve reperfusion. Sham operation group: No wire was inserted for ligation, and the rest of the treatment was the same as the drug administration group.

[0025] 2.3 Determination indicators 2.3.1 Neurological function score: The condition of the animals in each group was closely observed within 24 h of ischemia-reperfusion, and behavioral defects were evaluated with reference to the Zea Longa scoring standard, where 0 was normal with no symptoms of neurological damage; 1 was inability to fully extend the contralateral forelimb; 2 was turning in circles to the contralateral side when crawling; 3 was leaning to the contralateral side when walking; and 4 was inability to walk spontaneously and loss of consciousness.

[0026] 2.3.2 Cerebral infarction area: After 24 hours of reperfusion after drug administration, the animals in each group were killed, and the whole brain of half of the rats in each group was taken. The brain was sliced ​​coronally (2 mm) using a brain slice mold, and then the brain slices were placed in 2% triphenyltetrazolium chloride (TTC) solution at 37°C for 15 minutes in the dark, fixed with 4% paraformaldehyde for more than 2 hours, and photographed. Normal brain tissue appears red, and cerebral infarction tissue is not stained and appears white. The photos were analyzed using Image Pro Plus 6.0 software to calculate the infarction area. The infarction area% = infarction area / total brain area*100%.

[0027] 2.3.3 ELISA for detection of brain tissue TNF-α, IL-6, and IL-1β levels The whole brain of the remaining rats in each group was taken, weighed on a balance, and the tissue was made into a tissue homogenate with pre-cooled physiological saline (brain tissue g: physiological saline mL = 1:9), centrifuged at low temperature and high speed for 10 min, and the supernatant was taken. The detection was carried out according to the steps in the ELISA kit manual.

[0028] 2.4 Statistical analysis All experimental data were expressed as mean ± standard deviation (`X ± SD), and data between groups were compared using SPSS23.0 software, and one-way analysis of variance (ANOVA) was used for significance analysis.

[0029] 3. Results 3.1 Neurological function score and percentage of cerebral infarction area Compared with the sham operation group, the neurological function score and cerebral infarction area in the model group were significantly higher than those in the sham operation group; compared with the model group, the neurological function score and cerebral infarction area percentage in the two dose groups of nimodipine and colchicine were significantly reduced, as shown in Table 1.

[0030] Table 1 Comparison of neurological function scores and percentage of infarct area in rats of each group (`X±SD) Note: Compared with the sham operation group##P<0.01, compared with the model group*P<0.05**P<0.01 3.2 Contents of inflammatory factors TNF-α, IL-6, and IL-1β in brain tissue Compared with the sham operation group, the levels of inflammatory factors in the model group were higher than those in the sham operation group, with extremely significant differences; compared with the model group, the levels of inflammatory factors in both the nimodipine and colchicine dose groups were significantly reduced, with statistical differences. The reduction of inflammatory factors in the two colchicine dose groups was dose-dependent, as shown in Table 2 for details.

[0031] Table 2 Contents of inflammatory factors TNF-α, IL-6, and IL-1β in brain tissue of rats in each group (`X±SD) Note: Compared with the sham operation group##P<0.01, compared with the model group*P<0.05**P<0.01 The above test results show that colchicine can effectively improve neurological deficits, reduce the area of ​​cerebral infarction, reduce the degree of brain tissue edema, and reduce the level of inflammatory factors. It can prevent and relieve the symptoms of stroke from multiple angles and block the progression of the disease. It is safe and reliable, with no obvious toxic side effects.

[0032] The above is only a preferred embodiment of the present invention, and does not limit the present invention in any form. Although the present invention has been disclosed as a preferred embodiment as above, it is not used to limit the present invention. Any technician familiar with the present invention can make some changes or modify the technical contents suggested above into equivalent embodiments without departing from the scope of the technical solution of the present invention. However, any simple modification, equivalent change and modification made to the above embodiments according to the technical essence of the present invention without departing from the content of the technical solution of the present invention still fall within the scope of the solution of the present invention.

Claims

1. Use of colchicine as an effective ingredient for preventing and / or treating ischemic cerebrovascular disease in the preparation of a medicament for preventing and / or treating ischemic cerebrovascular disease.

2. The use according to claim 1, characterized in that The ischemic cerebrovascular disease is stroke, and the prevention is secondary prevention of stroke.

3. The use according to claim 2, characterized in that The stroke markers are highly expressed in the following markers: IL-1, IL-6, NLRP3, TNF-α or IL-8, inflammatory factors: A1AC (α1-antitrypsin), BPI (bactericidal / permeability-promoting protein), C5a (complement component 5a) and IL-1β 。 4. The use according to claim 3, characterized in that The stroke highly expresses the following markers: TNF-α, IL-6 and IL-1β.

5. The use of colchicine as claimed in claim 1, characterized in that In the medicine, colchicine is the only effective ingredient acting on ischemic cerebrovascular diseases.

6. Use of colchicine as an effective ingredient for preventing and / or treating ischemic cerebrovascular disease in the preparation of a medicament for treating patients with ischemic cerebrovascular disease, characterized in that: The patients also suffer from coronary heart disease, atrial fibrillation, hypertension, diabetes, obesity, chronic kidney disease and other diseases.

7. A pharmaceutical composition for preventing and / or treating ischemic cerebrovascular disease, characterized in that: The effective ingredient of the composition in the medicine for preventing and treating ischemic cerebrovascular diseases includes colchicine.

8. The pharmaceutical composition for preventing and / or treating ischemic cerebrovascular disease according to claim 7, characterized in that: The pharmaceutical composition uses colchicine as the only effective ingredient for preventing and treating ischemic cerebrovascular disease, wherein the ischemic cerebrovascular disease is cerebral stroke, and the prevention is secondary prevention of cerebral stroke.

9. The pharmaceutical composition for preventing and / or treating ischemic cerebrovascular disease according to claim 8, characterized in that: The pharmaceutical composition for preventing and / or treating ischemic cerebrovascular disease further comprises a pharmaceutically acceptable carrier.

10. The pharmaceutical composition for preventing and / or treating ischemic cerebrovascular disease according to claim 9, characterized in that: The pharmaceutical composition for preventing and / or treating ischemic cerebrovascular diseases is an injectable drug or an oral drug, and the form of the pharmaceutical composition for preventing and / or treating ischemic cerebrovascular diseases is selected from tablets, capsules, granules, suspensions, emulsions, solutions, syrups or injections.

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