CN113599502A - Composition of withaferin A and leptin and application of composition in preparation of medicine for preventing and/or treating hemorrhagic stroke - Google Patents
Composition of withaferin A and leptin and application of composition in preparation of medicine for preventing and/or treating hemorrhagic stroke Download PDFInfo
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- CN113599502A CN113599502A CN202110829521.8A CN202110829521A CN113599502A CN 113599502 A CN113599502 A CN 113599502A CN 202110829521 A CN202110829521 A CN 202110829521A CN 113599502 A CN113599502 A CN 113599502A
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- leptin
- withaferin
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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
- A61K31/56—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
- A61K31/58—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids containing heterocyclic rings, e.g. danazol, stanozolol, pancuronium or digitogenin
- A61K31/585—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids containing heterocyclic rings, e.g. danazol, stanozolol, pancuronium or digitogenin containing lactone rings, e.g. oxandrolone, bufalin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/22—Hormones
- A61K38/2264—Obesity-gene products, e.g. leptin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
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- 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
Abstract
The invention relates to a composition of withaferin A and leptin and application thereof in preparing a medicine for preventing and/or treating hemorrhagic stroke, belonging to the technical field of medicines. The invention provides a composition of withaferin A and leptin and application thereof in preparing a medicine for preventing and/or treating hemorrhagic stroke, wherein the existing withaferin A and leptin two compounds are optimally applied to the aspect of preparing the medicine for preventing and/or treating hemorrhagic stroke through combination, so that a new source is provided for preparing the medicine for preventing and/or treating hemorrhagic stroke, and the new medicinal value of withaferin A and leptin is developed. The technology of the invention is funded by Guangxi science and technology base and talent special fund (Guike AD 18281013) and Guangxi natural science fund youth fund (2018 GXNSFAA 138046) on the Guangxi natural science fund (2020 GXNSFAA 259036).
Description
Technical Field
The invention relates to a composition of withaferin A and leptin and application thereof in preparing a medicine for preventing and/or treating hemorrhagic stroke, belonging to the technical field of medicines.
Background
Hemorrhagic stroke, a sudden onset of blood circulation disorder, clinically manifested as transient or permanent dysfunction, is an important type of stroke, about 15%. The death rate of Chinese population cerebral hemorrhage accounts for about 61.7% of the death rate of cerebrovascular disease, and the high morbidity and mortality rate of the death rate become the leading death disease of Chinese. Although the pathophysiology research of cerebral hemorrhage has been advanced greatly, the effective treatment means is limited to surgical removal of hematoma, dehydration, reduction of cranial pressure and neuroprotection. From the aspect of curative effect, whether the operation treatment or the medicine intervention is carried out, the fatality rate of the cerebral hemorrhage is still high, and the recurrence rate, the complications and the sequelae are still many. Therefore, the search for new drugs, new targets and new mechanisms is extremely important for the treatment of hemorrhagic stroke.
WithaferinA (WFA) is a steroid lactone active ingredient with 28 carbons extracted and purified from the root of the natural plant withaferin. A series of studies also confirm that withaferin compounds show good neuroprotective effect in various nervous system diseases such as anxiety, cognitive disorder, insomnia, parkinson's syndrome, alzheimer's disease, stroke, nerve injury, etc. (laoauba et al, chinese pharmacological report 2018). Our previous studies also reported that withaferin a is effective in ameliorating inflammatory responses and apoptosis of vascular endothelial cells following brain trauma (julian et al, European journal of pharmacology 2020). The results all indicate that withaferin A can be a medicine for effectively preventing and treating hemorrhagic stroke. But has not been reported.
Leptin (LEP) is an important hormone synthesized and secreted mainly by adipocytes, mainly regulating food intake and energy metabolism balance, and also involved in many other physiological processes including immune response, oxidative stress, endocrine, immune and neural development, etc. The earliest crowd-queuing studies by So ̈ derberg, Stefan et al reported that leptin might be an important risk factor for the first hemorrhagic stroke (stroke, 1994). The increase in leptin levels is closely associated with a poor prognosis of cerebral hemorrhage. Exogenous administration of leptin in a hemorrhagic stroke model in mice can increase cerebral edema and neurological deficit, and the effect is considered to be that leptin promotes the activation of stat3 and further up-regulates the expression levels of inflammation-related factors such as COX-2 and iNOS (J Cereb Blood Flow Metab, 2013). Notably, in this study, administration of stat3 inhibitor after cerebral hemorrhage was not effective in improving cerebral edema, and the paradoxical results indicate that the role of leptin in cerebral hemorrhage is yet to be further resolved.
Hemorrhagic stroke has limited effective treatment means, and from the aspect of curative effect, the death rate of cerebral hemorrhage is still high, and the recurrence rate, complications and sequelae are still large no matter the cerebral apoplexy is treated by operation or intervened by medicines. Therefore, the search for new drugs, new targets and new mechanisms is extremely important for the treatment of hemorrhagic stroke.
Disclosure of Invention
The invention aims to provide application of a drug composition containing withaferin A/leptin in preparation of drugs for preventing and/or treating hemorrhagic stroke, wherein the withaferin A and the leptin can prevent and treat the hemorrhagic stroke under the condition that the mass ratio is (20 mg/kg: 4mg/kg), so that the respective dosage is obviously reduced, and the effect superior to that of single use can be achieved.
In order to achieve the above purpose, the invention provides the following technical scheme:
the invention provides a medicine composition containing withaferin A/leptin, which comprises withaferin A and leptin, wherein the mass ratio of the withaferin A to the leptin is 20mg/kg to 4 mg/kg.
The invention also provides application of the pharmaceutical composition in the technical scheme in preparation of drugs for preventing and treating hemorrhagic stroke.
The invention also provides application of the pharmaceutical composition in the technical scheme in preparation of a medicine for increasing the number of neural precursor cells.
The invention also provides application of the pharmaceutical composition in the technical scheme in preparation of medicines for treating neuroinflammation.
Preferably, the medicament takes withaferin A and leptin as active ingredients and also comprises pharmaceutically acceptable auxiliary materials or auxiliary ingredients.
Preferably, the dosage form of the medicament comprises one or more of tablets, powder, granules, capsules, oral liquid and sustained release agents.
Compared with the prior art, the invention has the beneficial effects that: the composition of withaferin A and leptin and the application thereof in preparing the medicine for preventing and/or treating the hemorrhagic stroke provided by the invention not only provide a new source for preparing the medicine for preventing and/or treating the hemorrhagic stroke, but also discover new medicinal values of withaferin A and leptin.
Drawings
FIG. 1 is a chemical structural diagram of withaferin A and leptin and a schematic diagram of the effects of withaferin A and leptin;
FIG. 2 is a statistical chart of the results of the bar-rotating test of the test mice in the control group and the hemorrhagic stroke model group induced by autologous blood injection, after 7 days of intraperitoneal injection of the control solvent, withaferin A (40 mg/kg), leptin (8 mg/kg) and withaferin A (20mg/kg) + leptin (4mg/kg), respectively, n =12,** P<0.01；
FIG. 3 is a schematic diagram and a statistical chart of footprint analysis of mice detected 7 days after intraperitoneal injection of control solvent, withaferin A (40 mg/kg), leptin (8 mg/kg) and withaferin A (20mg/kg) + leptin (4mg/kg) in a hemorrhagic stroke model group induced by body blood injection and a control group, respectively, wherein n =12,** P<0.01, * P<0.05；
FIG. 4 shows the number of neural precursor cells in lateral ventricle area of mice in the hemorrhagic stroke model group induced by body blood injection and the control group 7 days after intraperitoneal injection of control solvent, withaferin A (40 mg/kg), leptin (8 mg/kg) and withaferin A (20mg/kg) + leptin (4mg/kg), respectively, n =6,** P<0.01, * P<0.05；
FIG. 5 shows the level of Iba1 positive cells in the peripheral area of bleeding of mice in the hemorrhagic stroke model group induced by autologous blood injection and the control group, after 7 days of intraperitoneal injection of control solvent, withaferin A (40 mg/kg), leptin (8 mg/kg) and withaferin A (20mg/kg) + leptin (4mg/kg), respectively, n =6,** P<0.01, * P<0.05。
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and the following embodiments are used for illustrating the present invention and are not intended to limit the scope of the present invention.
The invention provides a medicine composition containing withaferin A/leptin, wherein the mass ratio of withaferin A to leptin is 20 mg/kg: 4mg/kg (the chemical structural formula of withaferin A and leptin is shown in figure 1).
The invention also provides application of the pharmaceutical composition in the technical scheme in preparation of drugs for preventing and treating hemorrhagic stroke. In the invention, the medicament preferably takes withaferin A and leptin as active ingredients and also comprises pharmaceutically acceptable auxiliary materials or auxiliary ingredients. In the invention, the dosage form of the medicament preferably comprises one or more of tablets, powder, granules, capsules, oral liquid and sustained release agents. The dosage of withaferin A and leptin in the preparation is not particularly limited, and the dosage of the drug in the conventional preparation is adopted. The invention has no special limitation on the types and contents of the auxiliary materials or auxiliary components used by the preparation formulation, and the preparation method can be realized by adopting a conventional method.
The sources of withaferin A and leptin are not particularly limited, and the withaferin A and leptin can be prepared from conventional commercial products.
Example 1
The mice used in the following examples were: male SPF grade C57BL/6 mice 7-8 weeks old, purchased from changshafeke laboratory animals llc, production license number: SCXK (Xiang) 2016-. Animals are raised in the center of SPF-level experimental animals in hospitals of Guilin medical college, the temperature of the experimental animal room is 20-26 ℃ (the daily temperature difference is less than or equal to 4 ℃), the humidity is 40-70%, and the illumination is 12h, and the light and shade are alternated.
Establishing a mouse hemorrhagic stroke model: male C57BL/6 mice were anesthetized by intraperitoneal injection of 10% chloral hydrate (40 mg/kg). After anesthesia, a mouse is placed on a brain stereotaxic apparatus for fixation, and a needle insertion point of a right lateral brain basal ganglia is positioned according to a brain atlas of the mouse, wherein the specific position is from the front cranium, the front and back are-0.0 mm, the left and right are + 1.0 mm, the depth is as follows: -3.0 mm. Drilling to dura mater, after the tail of a mouse is unhaired, rubbing with alcohol to expand a tail blood vessel, pressing the root of the tail to prevent blood from flowing back to fill the tail artery blood, cutting the tail artery with a sterilized surgical blade, after dropping the blood on a piece of wax paper, drawing the arterial blood of about 20 mu l with a micro-sampling needle, injecting blood at a uniform speed of 2 mu l/min, after keeping the needle for 5min after injecting 10 mu l, upwards pulling out the needle for about 0.5 mm, continuing to inject the rest arterial blood, and then withdrawing the needle after keeping the needle for 10 min. Bone wax seals the bone void and sutures the scalp. The control group was treated in the same manner by injecting the same amount of physiological saline from the right basal ganglia and the rest was treated in the same manner.
Animal grouping: the experimental model comprises a sham operation group, a cerebral hemorrhage model group, a model + withaferin A (40 mg/kg) group, a model + leptin (8 mg/kg) group, a model + withaferin A (20mg/kg) + leptin (4mg/kg) group, and mice are randomly distributed according to the groups after cerebral hemorrhage modeling.
And detecting the change condition of the behavioural indexes of each group 7 days after the intraperitoneal injection of each group of the medicines and the control solvent. And (3) detection of the behavioral indexes: after the cerebral hemorrhage model is established, the test is carried out on the 7 th day of the injection of each group of medicines, the mouse ethological index test exercise is carried out in a quiet and proper environment, and the test is carried out according to the relevant ethological standard.
Rolling rod experiment
The mice were moved to the test room and habituated for 1 h. The mice were placed on the test instrument (rotating rod) at an initial speed of 4rpm, gradually accelerated, and no further acceleration was observed after increasing the speed to 40rpm at 5 min. The time the mouse dropped from the rod was recorded. The same batch of mice was tested 3 times a day, each time at 20min intervals, for three consecutive days, and the average was taken. Wherein, compared with the normal group, the behavioral indexes of the model group are obviously reduced, and obvious dyskinesia appears; the withaferin A + leptin combination can remarkably recover the mouse motor dysfunction caused by cerebral hemorrhage, and the effect is superior to that of a single administration group (the result is shown in figure 2).
Footprint analysis experiment
The mice were moved to the test room and habituated for 1 h. The mice are placed on a full-automatic footprint analysis detector, and the detection is carried out after parameters are set. The same batch of mice was tested 3 times per day, each time at 20min intervals, and the average was taken. Wherein, compared with the normal group, the number of wrong steps of the model group is obviously increased, and obvious dyskinesia appears; the withanotin A + leptin combination group can obviously improve the mouse footprint movement disorder caused by cerebral hemorrhage, and the effect is superior to that of a single administration group (the result is shown in figure 3).
Number of neural precursor cells in lateral ventricle region of mouse
Neural precursor cells are key cells for promoting nerve regeneration of damaged areas after brain injury, and the number of the neural precursor cells shows the repair capacity after the brain injury. After the hemorrhagic stroke model is established, the quantity of the nerve precursor cells in the lateral ventricle area of each group of mice is detected by using an immunohistochemical technology 7 days after each group of drugs and a control solvent are injected in the abdominal cavity. The results were: compared with the normal group, the number of the lateral ventricle positive nerve precursor cells of the mice in the model group has no significant change. The withanotin A + leptin combination group can obviously increase the number of lateral ventricle positive nerve precursor cells, and the effect is superior to that of a single administration group (the result is shown in figure 4).
Inhibiting neuroinflammation after hemorrhagic apoplexy
The occurrence and development of neuroinflammation are considered to be one of the important factors for the progress of hemorrhagic stroke at present, and the role of neuroinflammation in promoting the mediation of neuron progressive death in the process of hemorrhagic stroke is widely proved. The occurrence of neuroinflammation mainly comprises inflammatory reactions such as microglial activation and the like. Furthermore, inflammatory cells in the center interact through a plurality of mechanisms to generate proinflammatory cytokines to amplify inflammatory signals and generate neurotoxin directly acting on neurons, and finally, hemorrhagic stroke injury is aggravated. After the hemorrhagic stroke model is established, the number of activated microglia in the hemorrhage peripheral area of each group of mice is detected by using an immunohistochemical technology after each group of drugs and a control solvent are injected into the abdominal cavity for 7 days. The results were: the number of activated microglia in the peripheral area of bleeding of the model group mice was significantly increased compared to the normal group. The withaferin A + leptin combination group can obviously reduce the number of activated microglia, and the effect is better than that of a single administration group (the result is shown in figure 5).
Claims (6)
1. The application of the withaferin A/leptin-containing pharmaceutical composition in preparing the medicines for preventing and/or treating the hemorrhagic stroke is characterized in that the optimal mass ratio of the withaferin A/leptin pharmaceutical composition is 20mg/kg to 4mg/kg, but not limited to the above ratio.
2. The use of the withaferin a/leptin containing pharmaceutical composition of claim 1 in the preparation of a medicament for preventing and or treating hemorrhagic stroke, wherein said withaferin a/leptin pharmaceutical composition ameliorates the neurological impairment following hemorrhagic stroke.
3. Use of the withaferin a/leptin containing pharmaceutical composition according to claim 1 for the preparation of a medicament for the prevention and or treatment of hemorrhagic stroke, characterized in that said withaferin a/leptin pharmaceutical composition increases the number of neural precursor cells.
4. The use of the withaferin a/leptin containing pharmaceutical composition of claim 1 in the preparation of a medicament for preventing and or treating hemorrhagic stroke, wherein said withaferin a/leptin pharmaceutical composition inhibits neuroinflammation.
5. The application of the withaferin A/leptin-containing pharmaceutical composition in preparing a medicine for preventing and/or treating hemorrhagic stroke as claimed in claim 1, wherein the withaferin A and leptin are used as active ingredients in the withaferin A/leptin pharmaceutical composition, and pharmaceutically acceptable auxiliary materials or auxiliary ingredients are also included.
6. The application of the withaferin A/leptin-containing pharmaceutical composition in preparing a medicine for preventing and/or treating hemorrhagic stroke as claimed in claim 1, wherein in the withaferin A/leptin pharmaceutical composition, the dosage form of the medicine comprises one or more of tablets, powder, granules, capsules, oral liquid and sustained release agents.
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103041371A (en) * | 2012-12-31 | 2013-04-17 | 上海市内分泌代谢病研究所 | Application of adiponectin in preparing medicine for curing cerebral arterial thrombosis of old people |
| US20160375039A1 (en) * | 2013-11-26 | 2016-12-29 | The Children's Medical Center Corporation | Compounds for the treatment of obesity and methods of use thereof |
| CN109985045A (en) * | 2019-05-10 | 2019-07-09 | 上海交通大学医学院附属瑞金医院 | Betulinic acid prepares the purposes for the treatment of leptin resistance related drugs as leptin sensitizer |
| CN109999044A (en) * | 2019-05-24 | 2019-07-12 | 北京大学 | A kind of composition containing withaferin A and its application |
| CN110403932A (en) * | 2019-09-05 | 2019-11-05 | 北京大学 | A kind of pharmaceutical composition and its application containing withaferin A |
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- 2021-07-22 CN CN202110829521.8A patent/CN113599502A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103041371A (en) * | 2012-12-31 | 2013-04-17 | 上海市内分泌代谢病研究所 | Application of adiponectin in preparing medicine for curing cerebral arterial thrombosis of old people |
| US20160375039A1 (en) * | 2013-11-26 | 2016-12-29 | The Children's Medical Center Corporation | Compounds for the treatment of obesity and methods of use thereof |
| CN109985045A (en) * | 2019-05-10 | 2019-07-09 | 上海交通大学医学院附属瑞金医院 | Betulinic acid prepares the purposes for the treatment of leptin resistance related drugs as leptin sensitizer |
| CN109999044A (en) * | 2019-05-24 | 2019-07-12 | 北京大学 | A kind of composition containing withaferin A and its application |
| CN110403932A (en) * | 2019-09-05 | 2019-11-05 | 北京大学 | A kind of pharmaceutical composition and its application containing withaferin A |
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