CN112587512A - ASBDV pharmaceutical composition for treating cerebral apoplexy and preparation and application thereof - Google Patents
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Abstract
The invention discloses an ASBDV pharmaceutical composition for treating cerebral apoplexy, and a preparation and application thereof. The ASBDV medicine composition for treating cerebral apoplexy consists of astaxanthin and cannabidiol. The preparation is the ASBDV pharmaceutical composition for treating cerebral apoplexy, and auxiliary materials are added into the ASBDV pharmaceutical composition to prepare tablets, oral liquid, capsules, granules, powder, pills or injections. The application is the application of the ASBDV pharmaceutical composition for treating cerebral apoplexy in preparing products for preventing/treating cerebral apoplexy. The astaxanthin and cannabidiol composition (ASBDV) has a remarkable effect of treating cerebral apoplexy.
Description
Technical Field
The invention belongs to the technical field of medicines, and particularly relates to an ASBDV pharmaceutical composition for treating cerebral apoplexy, and a preparation and application thereof.
Background
Cerebral apoplexy is a group of diseases which take cerebral ischemia and hemorrhagic injury symptoms as main clinical manifestations, is also called cerebral apoplexy or cerebrovascular accident, has extremely high fatality rate and disability rate, and is mainly divided into hemorrhagic cerebral apoplexy (cerebral hemorrhage or subarachnoid hemorrhage) and ischemic cerebral apoplexy (cerebral infarction and cerebral thrombosis). The cerebral apoplexy is acute and the fatality rate is high, which is one of the most important lethal diseases in the world. High mortality rate and gradually increased prevention and treatment cost become one of the most serious diseases harming the health of people, cause huge economic burden and mental stress to the society and families, and become an important public health problem seriously affecting the national civilization[1-2]Therefore, the research on effective therapeutic drugs is of great social significance.
Disclosure of Invention
The first purpose of the invention is to provide an ASBDV pharmaceutical composition for treating cerebral apoplexy; the second aim is to provide the preparation of the ASBDV pharmaceutical composition for treating cerebral apoplexy; the third purpose is to provide the application of the ASBDV pharmaceutical composition for treating cerebral apoplexy.
The first purpose of the invention is realized by that the ASBDV medicine composition for treating cerebral apoplexy consists of astaxanthin and cannabidiol.
The second purpose of the invention is realized by adding auxiliary materials into the ASBDV pharmaceutical composition to prepare tablets, oral liquid, capsules, granules, powder, pills or injections.
The third purpose of the invention is realized by the application of the ASBDV pharmaceutical composition for treating cerebral apoplexy in the preparation of products for preventing/treating cerebral apoplexy.
The present study shows that high intake of the six major carotenoids (i.e. lycopene, carotene, lutein, zeaxanthin and astaxanthin) is associated with a reduced risk of stroke and other cardiovascular events.
Astaxanthin (astaxanthin) is a xanthophyll carotenoid widely found in rhodococcus pluvialis, seafood, quail and some complex plants. It has antioxidant, antiinflammatory, anti-apoptosis, and neuroprotective effects. Some recent studies have shown that astaxanthin can prevent ischemia-reperfusion injury in mice by activating the mTOR pathway, exhibiting therapeutic effects on cerebral stroke.
Cannabidivarin (CBDV), the first isolated in 1969, has so far been poorly known for its pharmacological activity, CBDV has been shown to inhibit the major synthase of endocannabinoids, 2-arachidon, diacylglycerol lipase a, etc., whereas a study in 2018 has shown that chronic use of CBDV can restore behavioral patterns and brain atrophy symptoms in rats with rist syndrome (an autism), although these pharmacological traits have not been clinically proven, they illustrate the diversity of CBDV pharmacology. The invention proves the curative effect of the astaxanthin and cannabidiol combined drug (ASBDV) on cerebral arterial thrombosis for the first time.
Drawings
FIG. 1 is a graph showing the effect of the ASBDV pharmaceutical composition of the present invention on the survival rate of mice after stroke;
FIG. 2 is a graph showing the effect of the ASBDV pharmaceutical composition of the present invention on the body weight of mice after stroke;
FIG. 3 is a graph showing the effect of the ASBDV pharmaceutical composition of the present invention on mNSS of post-stroke mice;
fig. 4 is a schematic diagram showing the comparative effect of the ASBDV pharmaceutical composition of the present invention on the 5 th day movement distance of the post-stroke mouse in the field test;
FIG. 5 is a graph showing the comparative effect of the ASBDV pharmaceutical composition of the present invention on the mean velocity at day 5 of the post-stroke mouse field test;
fig. 6 is a schematic diagram showing the comparative effect of the ASBDV pharmaceutical composition of the present invention on the limb support distance at day 7 of the post-stroke mouse open field test;
fig. 7 is a graph showing the maximum contact area versus effect of the ASBDV pharmaceutical composition of the present invention on abnormal improvement in post-stroke mice;
fig. 8 is a graph showing the effect of the ASBDV pharmaceutical composition of the present invention on improving abnormal hind limb paw prints in post-stroke mice;
fig. 9 is a schematic diagram showing the effect of the ASBDV pharmaceutical composition of the present invention on brain injury in mice after stroke.
Detailed Description
The present invention is further illustrated by the following examples and the accompanying drawings, but the present invention is not limited thereto in any way, and any modifications or alterations based on the teaching of the present invention are within the scope of the present invention.
The ASBDV medicine composition for treating cerebral apoplexy consists of astaxanthin and cannabidiol.
The mass ratio of the astaxanthin to the cannabidiol is 1: (2-5).
The mass ratio of the astaxanthin to the cannabidiol is 1: 3.
The preparation of the ASBDV pharmaceutical composition for treating cerebral apoplexy is prepared by adding auxiliary materials into the ASBDV pharmaceutical composition to prepare tablets, oral liquid, capsules, granules, powder, pills or injections.
The application of the ASBDV pharmaceutical composition for treating cerebral apoplexy disclosed by the invention is the application of the ASBDV pharmaceutical composition for treating cerebral apoplexy in preparing a product for preventing/treating cerebral apoplexy.
The product is food, medicine or health food.
The invention is further illustrated by the following specific examples:
example 1
Weighing astaxanthin and water-soluble cannabidiol according to the mass ratio of 1:2, and then uniformly mixing to obtain the target ASBDV pharmaceutical composition for treating cerebral apoplexy.
Example 2
Weighing astaxanthin and water-soluble cannabidiol according to the mass ratio of 1:3, and then uniformly mixing to obtain the target ASBDV pharmaceutical composition for treating cerebral apoplexy.
Example 3
Weighing astaxanthin and water-soluble cannabidiol according to the mass ratio of 1: 4, and then uniformly mixing to obtain the target ASBDV pharmaceutical composition for treating cerebral apoplexy.
Example 4
Weighing astaxanthin and water-soluble cannabidiol according to the mass ratio of 1: 5, and then uniformly mixing to obtain the target ASBDV pharmaceutical composition for treating cerebral apoplexy.
Example 5
The ASBDV pharmaceutical composition for treating cerebral apoplexy prepared in the embodiment 2 is added with pharmaceutically acceptable auxiliary materials to prepare tablets.
Example 6
The ASBDV pharmaceutical composition for treating cerebral apoplexy prepared in the embodiment 1 is added with pharmaceutically acceptable auxiliary materials to prepare oral liquid.
Example 7
The ASBDV pharmaceutical composition for treating stroke prepared in example 4 is added with pharmaceutically acceptable excipients to prepare a capsule.
Example 8
The ASBDV pharmaceutical composition for treating cerebral apoplexy prepared in the embodiment 2 is added with pharmaceutically acceptable auxiliary materials to prepare granules.
Example 9
The ASBDV pharmaceutical composition for treating cerebral apoplexy prepared in the embodiment 3 is added with pharmaceutically acceptable auxiliary materials to prepare powder.
Example 10
The ASBDV medicine composition for treating cerebral apoplexy prepared in the embodiment 2 is added with pharmaceutically acceptable auxiliary materials to prepare pills.
Example 11
The ASBDV pharmaceutical composition for treating cerebral apoplexy prepared in the embodiment 3 is added with pharmaceutically acceptable auxiliary materials to prepare an injection.
Example 12
The ASBDV medicine composition for treating cerebral apoplexy prepared in the embodiment 4 is added with pharmaceutically acceptable auxiliary materials to prepare pills.
Example 13
A capsule is prepared by adding pharmaceutically acceptable auxiliary materials into the ASBDV pharmaceutical composition for treating cerebral apoplexy, which is prepared in the embodiment 1.
Example 14
The ASBDV pharmaceutical composition for treating stroke prepared in example 3 was tested as follows:
1. materials and methods
1.1 animals
Male C57BL/6J mice, 8 weeks old, purchased from the department of laboratory animals, university of Kunming medical, were 22.5-25 g in weight, kept in a room of fixed humidity (40% + -5%), temperature (22 deg.C + -2 deg.C) and natural light, and had free access to water and food. The animal study protocol was approved by the university of Kunming medical laboratory animal ethics Committee.
1.2 establishment of cerebral ischemia-reperfusion injury model and group administration
Mice were anesthetized with a small animal anesthesia machine filled with 2% isoflurane, the skin of the mouse head was cut open, and the basal cerebral blood flow was monitored with a laser doppler flow meter (Moor Instruments, uk). The left common carotid, external carotid, internal carotid and pterygopalatine arteries of the mice were isolated, the distal external carotid was ligated with medical sutures, the proximal cut was made and a MCAO plug was inserted to block blood flow in the middle cerebral artery. And after 30 min, the plug is drawn out to realize reperfusion, and the mice which detect the cerebral blood flow until the cerebral blood flow is recovered to about 80 percent of the basic value are considered to realize the purpose of ischemia reperfusion, so that the modeling is successful. Finally, the skin was sutured with 4-0 sutures. The sham group (sham) was identical to the model rats except that no wire plugs were inserted. 140 mice were used for this study, 20 of which were discarded without effecting reperfusion of blood flow, and 120 were equally divided for the official experiment in 6 groups as follows: sham group (sham), model group (model), 3 mL. kg-1Gold multiple control drug group (ginaton), 3 mL. kg-1 ASBDV pharmaceutical composition group (ASBDV) for treating cerebral apoplexy, prepared in example 3, 3 mL kg-1 Astaxanthin group (AST) and 3 mL kg-1 time cannabidiol group (CBDV). For 4 administration groups, immediately after moldingThe corresponding drug was injected into the tail vein 1 time, and thereafter, the tail vein was administered 1 time in the morning and evening each day until day 7, and the Sham (Sham) and Model (Model) groups were administered with an equal amount of physiological saline.
Neurological deficit scores mice were scored for neurological deficit on days 1, 4, and 7 following cerebral ischemia-reperfusion according to a modified neurological severity score sheet (mNSS) including 4 items of tail overhang, walking, balance beam test, and reactive deficit. The score ranges from 0 to 14 and is positively correlated with the severity of ischemia reperfusion injury.
Open field experiments on day 5 after ischemia reperfusion, mice were placed in a 40 cm x 40 cm open field with a high speed camera suspended above. To eliminate the odor effect, the open field bottom was wiped with 75% alcohol before testing each animal. The recording time was set to 30 min and the total distance and average speed were recorded using ANY ANY-maze behavior tracking software (Stoelting, USA).
Automatic gait analysis mice were trained on the Catwalk system (Noldus, the netherlands) race for 2 days prior to ischemia reperfusion to become familiar with the race track, and on day 7 after ischemia reperfusion, mice were subjected to automatic quantitative gait recordings by the Catwalk system and analyzed for support distance, maximum contact area, total area of paw prints, stride length, and swing speed using Catwalk XT Version 9.1 software.
Measurement of cerebral infarction volume mice were euthanized and the brains were removed and cut into 5 coronal sections 2 mm thick in a brain mold and stained with TTC for 15 min in a 37 ℃ incubator. The areas of the contralateral hemisphere (Ci), ipsilateral hemisphere (Ii) and ipsilateral non-ischemic region (Ni) were determined using Image J software (NIH) and infarct volume (%) was calculated using the following equation:
blood Brain Barrier (BBB) leakage evaluation after stroke on the 6 th day according to 300mL kg-1 tail vein injection of 0.6% Evans blue solution, after 24 h taking the brain and cut into 5 2 mm thick coronary slices, using IVIS LuminaK Series III small animal in vivo optical imaging system (according to the following fluorescence imaging parameters uptake total luminous efficiency), exposure time automatic, excitation light 700 nm, emission light 790 nm.
Analysis of cerebral edema after inhalation of anesthetized mice with 2% isoflurane, 15 mL kg-1Tail vein injection of contrast agent iohexol injection. The brain was then scanned using the Quantum FX small animal in vivo three-dimensional tomography system (Micro-CT, PerkinElmer, usa) at the following parameters: the voltage is 90 kV, the current is 180 muA, the visual field is 2 cm, and the scanning time is 4.5 min.
2. Results
2.1 Effect of each administration group on survival, body weight and mNSS in post-stroke mice
As can be seen from FIG. 1, the mortality rate was higher (P < 0.001) in the Model group mice (Model) than in the Sham group (Sham). The ASBDV pharmaceutical composition for treating stroke (ASBDV) (P < 0.001) prepared in example 3 improved survival rate to more than 50% and the hypocannabidiol (CBDV) and Astaxanthin (AST) groups were slightly less effective (P < 0.05), compared to the Model group (Model), and the survival rate was improved after treatment with ASBDV, Astaxanthin (AST) and hypocannabidiol (CBDV) prepared in example 3. As can be seen from fig. 2, the body weight of the model group was significantly reduced, and compared to the model group, the ASBDV pharmaceutical composition for treating stroke prepared in example 3 exhibited significant efficacy (P < 0.05 or P < 0.01) from day 4, and Cannabidiol (CBDV) and Astaxanthin (AST) were effective (P < 0.01) from day 5, and on the mNSS scale (fig. 3), the model group mice exhibited severe impaired neurobehavioral (P < 0.001), and on day 4 after surgery, the ASBDV pharmaceutical composition for treating stroke (asv) (P < 0.001) and Cannabidiol (CBDV) (P < 0.05) prepared in example 3 significantly improved neurological abnormalities, and this beneficial effect was sustained until day 7 (P < 0.001 or P < 0.01). The ASBDV pharmaceutical composition (ASBDV) for treating cerebral apoplexy prepared in the example 3 and the gold poly (Ginaton) as a positive drug have no obvious difference on the 3 indexes.
2.2 Effect of each administration group on post-stroke mouse locomotor function
Open field experiments (fig. 4 and 5) on day 5 after stroke show that the moving distance and the average speed of the model group mice are obviously reduced within 30 min (P< 0.01). However, the ASBDV pharmaceutical composition for treating cerebral Apoplexy (ASBDV) prepared in example 3 was compared with the control drug group (Ginanton) (P)<0.001), Astaxanthin (AST) group and cannabidiol sub (CBDV) group (B: (B)P Less than 0.05) the moving distance and the average speed of the mouse are obviously improved. Gait analysis of mice on day 7 showed that the ASBDV pharmaceutical composition (ASBDV), Astaxanthin (AST) and Cannabidiol (CBDV) groups for treating stroke prepared in example 3 could significantly improve the base of support (BOS) (P) caused by stroke<0.001, fig. 6), and in addition, the ASBDV pharmaceutical composition for treating cerebral Apoplexy (ASBDV) prepared in example 3 can improve the maximum contact area of abnormality (P < 0.05) and Total area of paw prints of hind limbs: (P Less than 0.01), the effect is obviously better than that of Astaxanthin (AST) group and Cannabidiol (CBDV) group (BP< 0.05, FIGS. 7, 8). In the above kinematic evaluation indexes, the ASBDV pharmaceutical composition for treating cerebral Apoplexy (ASBDV) prepared in example 3 is equivalent to the gold pleiotropium positive drug.
2.3 Effect of Each administration group on post-stroke brain injury in mice
Mice were evaluated for brain injury severity by TTC staining and detection of BBB leakage. The model group mice had significantly increased cerebral infarct volume (A, B of fig. 9) and BBB leakage (C, D of fig. 9) compared to Sham (Sham) ((ii) in contrast to Sham)P< 0.001). Example 3 the resulting pharmaceutical composition of ASBDV for stroke treatment (ASBDV) can reverse this brain injury: (P< 0.001 andP< 0.01). While the Cannabidiol (CBDV) and Astaxanthin (AST) groups also reduced the volume of cerebral infarction, the ASBDV pharmaceutical composition for treating cerebral Apoplexy (ASBDV), Cannabidiol (CBDV) and Astaxanthin (AST) groups prepared in example 3 all had a vague effect in terms of BBB leakage, showing a beneficial effect. In addition, the preparation of example 3 for the treatment of strokeASBDV pharmaceutical composition (ASBDV) and gold were much the same in reversing cerebral infarct volume and BBB leakage.
3. Conclusion
The ASBDV pharmaceutical composition (ASBDV) for treating stroke prepared in example 3 has a significant effect on post-stroke cerebellar injury, and the ASBDV pharmaceutical composition (ASBDV) (P < 0.001) for treating stroke prepared in example 3 can increase the survival rate to more than 50% and show a significant effect on the fourth day (fig. 1, 2 and 3).
ASBDV pharmaceutical composition for treating cerebral Apoplexy (ASBDV) prepared in example 3 in improving motor function<0.001), Astaxanthin (AST) group and cannabidiol sub (CBDV) group (B: (B)P < 0.05) the moving distance and average speed of the mice were significantly increased (fig. 4, 5).
Gait analysis of mice shows that the ASBDV pharmaceutical composition (ASBDV), the Astaxanthin (AST) group and the Cannabidiol (CBDV) group for treating stroke prepared in example 3 can significantly improve the base of support (BOS) (P) caused by stroke<0.001, 6), and in addition, the ASBDV pharmaceutical composition group for treating cerebral Apoplexy (ASBDV) prepared in example 3 can improve the abnormal maximum contact area ((ASBDV)P< 0.05) and Total area of paw prints of hind limbs: (PLess than 0.01), the effect is obviously better than that of Astaxanthin (AST) group and Cannabidiol (CBDV) group (BP< 0.05, FIGS. 7, 8). In the above kinematic evaluation indexes, the ASBDV pharmaceutical composition for treating cerebral Apoplexy (ASBDV) prepared in example 3 is equivalent to the gold pleiotropium positive drug.
The severity of brain injury to mice is evaluated by TTC staining and BBB leakage detection, and the results show that the ASBDV pharmaceutical composition (ASBDV), Cannabidiol (CBDV) and Astaxanthin (AST) groups for treating cerebral apoplexy prepared in example 3 all have obvious efficacy, and the ASBDV pharmaceutical composition (ASBDV) for treating cerebral apoplexy prepared in example 3 has more obvious effect, can obviously reverse the brain injury, and is superior to the Cannabidiol (CBDV) and Astaxanthin (AST) groups in results (figure 9).
Example 15
The results of experiments carried out by the ASBDV pharmaceutical compositions for treating cerebral apoplexy prepared in example 1, example 2 and example 4 are the same as example 14, and all the results show that the ASBDV pharmaceutical compositions for treating cerebral apoplexy have significant efficacy in treating cerebral apoplexy.
Claims (6)
1. The ASBDV pharmaceutical composition for treating cerebral apoplexy is characterized by consisting of astaxanthin and cannabidiol.
2. The ASBDV pharmaceutical composition for treating cerebral apoplexy according to claim 1, wherein the weight ratio of astaxanthin and cannabidivarin is 1: (2-5).
3. The ASBDV pharmaceutical composition for treating cerebral apoplexy according to claim 1, wherein the weight ratio of the astaxanthin to the cannabidiol is 1: 3.
4. The preparation of the ASBDV pharmaceutical composition for treating the cerebral apoplexy as claimed in any one of claims 1 to 3, wherein the preparation of the ASBDV pharmaceutical composition for treating the cerebral apoplexy is prepared by adding auxiliary materials into the ASBDV pharmaceutical composition to prepare tablets, oral liquid, capsules, granules, powder, pills or injections.
5. The application of the ASBDV composition for treating cerebral apoplexy of any claim 1 to 3, which is characterized in that the ASBDV composition for treating cerebral apoplexy is applied to the preparation of products for preventing/treating cerebral apoplexy.
6. The use of ASBDV composition for the treatment of stroke as claimed in claim 5 wherein the product is a food, pharmaceutical or nutraceutical product.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101361726A (en) * | 2008-09-05 | 2009-02-11 | 南通大学 | Application of astaxanthin in the preparation of drugs for the prevention and treatment of stroke |
| CN101489548A (en) * | 2006-06-08 | 2009-07-22 | 纽若科伊公司 | Use of cannabinoid receptor agonists as hypothermia inducing drugs for the treatment of ischemia |
| US20140228438A1 (en) * | 2011-07-01 | 2014-08-14 | Gw Pharmaceuticals Limited | Cannabinoids for use in the treatment of neurodegenerative diseases or disorders |
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2021
- 2021-02-05 CN CN202110162171.4A patent/CN112587512A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101489548A (en) * | 2006-06-08 | 2009-07-22 | 纽若科伊公司 | Use of cannabinoid receptor agonists as hypothermia inducing drugs for the treatment of ischemia |
| CN101361726A (en) * | 2008-09-05 | 2009-02-11 | 南通大学 | Application of astaxanthin in the preparation of drugs for the prevention and treatment of stroke |
| US20140228438A1 (en) * | 2011-07-01 | 2014-08-14 | Gw Pharmaceuticals Limited | Cannabinoids for use in the treatment of neurodegenerative diseases or disorders |
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Application publication date: 20210402 |