CN110664809B - Pharmaceutical composition for preventing and treating cerebral infarction - Google Patents
Pharmaceutical composition for preventing and treating cerebral infarction Download PDFInfo
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- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
- A61K31/4353—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems
- A61K31/4375—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems the heterocyclic ring system containing a six-membered ring having nitrogen as a ring heteroatom, e.g. quinolizines, naphthyridines, berberine, vincamine
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- A61K31/00—Medicinal preparations containing organic active ingredients
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- 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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Abstract
The invention relates to a pharmaceutical composition, which consists of phellodendrine, paeoniflorin and optional EGCG. The pharmaceutical composition can prevent or treat brain tissue damage caused by local and whole cerebral ischemia, specifically can prevent or treat diseases such as cerebral hemorrhage, cerebral infarction, cerebral thrombosis and the like, particularly has a very good treatment effect on cerebral infarction, and has high safety and no side effect.
Description
Technical Field
The invention relates to the field of medicines, in particular to a pharmaceutical composition containing phellodendrine, paeoniflorin and optionally EGCG, a preparation method and application of the pharmaceutical composition in preventing and treating cerebral infarction.
Background
Cerebral infarction, also known as ischemic stroke, refers to ischemic necrosis or softening of localized brain tissue caused by blood supply disorder, ischemia, and hypoxia in the brain. The common clinical types of cerebral infarction include cerebral thrombosis, lacunar infarction, cerebral embolism and the like, and the cerebral infarction accounts for 80% of all cerebral apoplexy; the clinical manifestations are characterized by sudden syncope, unconsciousness, hemiplegia, speech disturbance and intellectual disturbance. Cerebral infarction not only poses great threat to human health and life, but also brings great pain and heavy burden to patients, families and society.
After cerebral infarction is caused, under the action of various pathophysiological mechanisms, neuronal cell necrosis is caused, and finally, the neurological function of a patient is damaged. The conventional treatment scheme of cerebral infarction generally only aims at a single link or a single factor of the pathological process of cerebral infarction, but the pathological process of cerebral infarction is extremely complex, and theoretically, the intervention of a plurality of links can achieve better curative effect.
The incidence of cerebral infarction in China is rising year by year and the onset age is in a decreasing trend, and the results of 10-year collaborative research on the world's biggest cardiovascular disease published by the world health organization show that the incidence of cerebral apoplexy in China is 250/10 ten thousand, is second only in the Siberian area (300/10 thousand) and second in the world, is a common disease and frequently encountered disease in China, harms the health of people and brings heavy mental and economic burden to the families of patients, so that the search of an economical and effective treatment scheme becomes urgent.
At present, more medicines are used for treating cerebral infarction, and particularly, traditional Chinese medicines are more, but the treatment effect is general and the effect is slow. For example, the prior art CN109731084A discloses a traditional Chinese medicine composition for treating stroke and cerebral infarction, which is prepared from the following raw materials in parts by weight: 12-16 parts of angelica sinensis, 12-18 parts of pseudo-ginseng, 7-13 parts of scorpion, 18-22 parts of peach kernel, 7-15 parts of radix puerariae, 5-10 parts of rhizoma corydalis, 8-12 parts of rhizoma sparganii, 8-12 parts of astragalus membranaceus, 15-25 parts of radix achyranthis bidentatae, 1-9 parts of leech, 8-12 parts of schizonepeta and 7-13 parts of fructus aurantii.
CN108714169A discloses a medicine for treating acute cerebral infarction, which is prepared from the following raw materials in parts by weight: 0.3-0.45 part of musk; 120 portions of astragalus root and 180 portions; 0.5-0.75 part of borneol; 6-9 parts of angelica; 6-9 parts of red peony root; 3-4.5 parts of peach kernels; 3-4.5 parts of safflower; 3-4.5 parts of ligusticum wallichii; 3-4.5 parts of earthworm.
Therefore, there is a need for a drug which can prevent or treat brain tissue damage caused by local and global cerebral ischemia, particularly, diseases such as cerebral hemorrhage, cerebral infarction and cerebral thrombosis, and which is highly safe and free from side effects.
Disclosure of Invention
Aiming at the defects in the prior art, the invention provides a pharmaceutical composition which can prevent or treat brain tissue damage caused by local and whole cerebral ischemia, particularly can prevent or treat diseases such as cerebral hemorrhage, cerebral infarction, cerebral thrombosis and the like, has a very good treatment effect on cerebral infarction, and has high safety and no side effect.
One of the objects of the present invention is to provide a pharmaceutical composition, which is characterized in that: it is composed of phellodendrine and paeoniflorin, and the weight ratio of phellodendrine to paeoniflorin is 0.1-1: 1-10.
Preferably, the weight ratio of phellodendrine to paeoniflorin is 0.5-1:1-5, preferably 1: 5.
Preferably, the pharmaceutical composition further comprises EGCG.
Preferably, the weight ratio of phellodendrine, paeoniflorin and EGCG is 0.1-3:1-10: 0.5-5.
Preferably, the weight ratio of phellodendrine, paeoniflorin and EGCG is 0.5-2:3-6:0.5-3, preferably 1:5: 1.
Phellodendrine is one of characteristic components and important active components of phellodendron amurense of Rutaceae, has obvious pharmacological activity, and modern pharmacology shows that the phellodendrine has obvious effects on the aspects of reducing blood pressure, resisting nephritis, inhibiting cellular immune response, inhibiting central nervous system and the like.
Paeoniflorin, also known as peoniflorin, is a pinane monoterpene picrin isolated from the traditional Chinese medicines radix Paeoniae Rubra and radix Paeoniae alba. Is a hygroscopic amorphous powder. Is present in root of Paeonia plant such as Paeonia lactiflora, Paeonia suffruticosa, etc. Has tranquilizing, analgesic, and antiinflammatory effects. Has antipyretic and spasmolytic effects, and can dilate coronary artery and increase coronary blood flow. Resisting acute myocardial ischemia, inhibiting blood platelet coagulation, and lowering blood pressure. Especially has the effect of protecting cerebral ischemia, and the paeoniflorin has the influence on blood brain barrier and cerebral ischemia neuropathological change symptoms and cerebral blood flow during reperfusion after cerebral ischemia. The paeoniflorin is found to have protective effect on cerebral edema, blood brain barrier and cerebral local blood flow after cerebral ischemia. The application of paeoniflorin in animals with focal cerebral ischemia can obviously relieve cerebral edema, reduce cerebral infarction range, improve neurobehavioral symptoms, improve blood brain barrier permeability, and increase cerebral local blood flow.
EGCG (epigallocatechin gallate), is the main component of green tea polyphenol, is catechin monomer separated from tea, and has antibacterial, antiviral, antioxidant, arteriosclerosis resisting, thrombosis resisting, vascular proliferation resisting, antiinflammatory and antitumor effects.
Preferably, the composition further optionally comprises a pharmaceutically acceptable carrier.
A pharmaceutically acceptable carrier refers to a pharmaceutically acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, manufacturing aid (e.g., lubricant, talc magnesium, calcium or zinc stearate, or stearic acid), or solvent encapsulating material, involved in transporting or transporting the subject compound from one organ or portion of the body to another organ or portion of the body. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not injurious to the patient. Some examples of materials that can be used as pharmaceutically acceptable carriers include: (1) sugars such as lactose, glucose and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose and its derivatives, such as microcrystalline cellulose, sodium carboxymethylcellulose, ethyl cellulose and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository waxes; (9) oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; (10) glycols, such as propylene glycol; (11) polyols such as glycerol, sorbitol, mannitol and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffering agents such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) ringer's solution; (19) ethanol; (20) a pH buffer solution; (21) polyesters, polycarbonates and/or polyanhydrides; and (22) other non-toxic compatible substances used in pharmaceutical formulations.
Another object of the present invention is to provide a method for preparing the composition, which comprises the steps of: the composition is prepared by uniformly mixing phellodendrine, paeoniflorin, EGCG and a pharmaceutically acceptable carrier according to the weight ratio.
The third purpose of the invention is to use the pharmaceutical composition in preparing a medicament for preventing or treating brain tissue damage caused by local and whole brain ischemia.
Preferably, the pharmaceutical composition is used for preparing a medicament for preventing or treating cerebral hemorrhage, cerebral infarction and cerebral thrombosis.
Preferably, the pharmaceutical composition is used for preparing a medicament for preventing or treating cerebral infarction.
In the case of pharmaceutical compositions for oral, sublingual, subcutaneous, intramuscular, intravenous, transdermal, topical or rectal administration, the active ingredient, alone or together with other active ingredients, may be mixed with conventional pharmaceutical carriers for administration to animals or humans in the form of administration unit dosage forms. Suitable unit dosage forms for administration include oral forms such as tablets, capsules, pills, powders, granules, chewing gum forms and oral solutions or suspensions, topical forms, transdermal forms, subcutaneous forms, intramuscular forms, intravenous forms or intraocular and rectal forms.
When preparing solid compositions in tablet form, a wetting agent, such as sodium lauryl sulfate, may be added to the micronized or unmicronized active ingredient and mixed with pharmaceutically acceptable excipients, such as silicon dioxide, starch, lactose, magnesium stearate, talc or the like. The tablets may be coated with sucrose, various polymers or other suitable materials, or otherwise treated to provide prolonged or sustained activity and continuous release of a predetermined amount of the active ingredient.
Capsule preparations are obtained by mixing one or more active ingredients with a diluent, such as ethylene glycol or glycerol esters, and then adding the resulting mixture to soft or hard gelatin capsules.
The preparation in the form of a syrup or spirit may contain one or more active ingredients with the addition of sweeteners, preferably non-caloric sweeteners, methylparaben, propylparaben as preservatives, as well as flavoring agents and suitable coloring agents.
Powders or granules which may be dispersed in water may contain one or more active ingredients in admixture with a dispersing or wetting agent, or suspending agent such as polyvinylpyrrolidone, or with a sweetening or taste-modifying agent.
The pharmaceutical composition can prevent or treat brain tissue damage caused by local and whole cerebral ischemia, specifically can prevent or treat diseases such as cerebral hemorrhage, cerebral infarction, cerebral thrombosis and the like, particularly has a very good treatment effect on cerebral infarction, and has high safety and no side effect.
Detailed Description
The present invention is illustrated in detail by the following examples, which are intended to be exemplary only.
It should be understood that the terms or words used in the specification and claims should not be construed as having meanings defined in dictionaries, but should be interpreted as having meanings that are consistent with their meanings in the context of the present invention on the basis of the following principles: the concept of terms may be defined appropriately by the inventors for the best explanation of the invention.
EXAMPLE 1 protective Effect of pharmaceutical composition on model of hypoxia and hypoglycaemia of neural cells
Cell culture: recovering frozen PC12 cell strain in centrifuge tube, centrifuging to obtain supernatant, suspending with 10% FBS-containing RPMI1640 culture medium, inoculating in culture dish, culturing for passage with incubator, harvesting cells at logarithmic growth phase, suspending with culture medium to adjust concentration to 1 × 104One/ml was plated in 24-well plates for subsequent testing.
Grouping experiments: normal control group, continuously administered with 95% O2And 5% CO2The mixed gas of (3); the second group is the sugar-deficient hypoxia (OGD) model group: the normal medium was removed from PC12 cells and replaced with HBSS solution (140mM NaCl, 3.5mM KCl, 12mM MgSO) containing no glucose4，5mM NaHCO3,1.7mM CaCl2,0.4mM KH2PO410mM Hepes) into a dedicated anoxic tank, charged with 95% N2And 5% CO2The mixed gas is lack of sugar and oxygen for 6 hours; in the treatment group, after the cells were pretreated with DMEM solution containing different pharmaceutical compositions for 3h, the cells were subjected to the OGD model for 6 h.
Detection indexes are as follows:
1. the cell viability of each group was determined by MTT method, and was ═ control OD value-other group OD value)/control OD value × 100%.
2. And (3) carrying out synergistic action judgment by adopting a gold positive average q value method, wherein the q value is obtained by the following formula: q ═ PA+B/(PA+PB-PA×PB). In the formula PA、PBAnd PA+BRespectively the treatment rates of the A medicine group, the B medicine group and the combination of the two medicines. q < 1 > sayThe two medicines can generate antagonism after being used together; q is more than 1, which indicates that the two drugs produce synergistic effect after being combined together, and q is 1, which indicates that the two drugs produce additive effect after being combined together.
3. LDH and SOD activity determination, wherein a biochemical kit of Nanjing institute of bioengineering is adopted for determination on a full-automatic biochemical instrument, and the specific detection refers to the kit specification.
The specific results are as follows:
1. the survival rate of each group of cells and the q value condition are determined by adopting an MTT method
# represents p <0.01 compared to normal control by Oneway-ANOVA test; represents p <0.01 compared to model group.
2. LDH and SOD Activity assay
From the results, the medicinal composition of phellodendrine and paeoniflorin has a very good protective effect on an oxygen-deficient and sugar-deficient model of nerve cells, and a synergistic effect is generated, so that the effect of the medicinal composition can be further improved after EGCG is added.
Example 2: effect of drugs on cerebral ischemia reperfusion model in rats
Test materials
Animals: healthy Wistar rats are domesticated in an animal feeding room for more than 3 days, are all male, and have the weight of 180-220 g, and the total number is 80.
Test method
The right carotid artery was dissected by median incision under isoflurane anesthesia (3% at the time of introduction and 2% at the time of maintenance) using an anesthetic Wistar rat, and blood flow in the common and external carotid arteries was temporarily blocked after they were placed on the line. After the external carotid artery was cut by inserting a plug embolus from the external carotid artery, 10.5mm was inserted into the internal carotid artery from the bifurcation of the external carotid artery and the internal carotid artery, and the surgical wound was closed after closing the initial part of the middle cerebral artery. The heating pad is used in the operation for cerebral infarction, and the rectal temperature of the mouse is kept at 36.5-37.5 ℃. The mice in ischemia were returned to cages for free rearing. After 30 minutes of infarction, the plug was removed under anesthesia and the incision was closed.
Animal grouping and dosing experiments: animals were randomly divided into 8 groups of 10 animals each, and the total was divided into a sham operation group, a model group and a drug group. The test substance was administered 2 times in total after 6 and 24 hours of ischemia reperfusion. The administration solution was filled in a 1mL syringe and administered with a 27G needle in an amount of 0.2mL from the tail vein. The sham operation group and the model group were injected with physiological saline of equal volume, respectively.
Measurement of cerebral infarction region:
after 2 days of ischemia reperfusion, mice were sacrificed by cutting off abdominal aorta under anesthesia for exsanguination, and brains were collected. The collected brains were prepared as 4 transverse brain sections 2mm thick from the border between the brain and the cerebellum using a mouse brain microtome. They were stained with 1% 2,3, 5-triphenyltetrazolium chloride (TTC) at 37 ℃ for 30 minutes. Brain sections were photographed using a microscope, and the left brain area, right brain non-obstacle (lesion) area, and right brain obstacle area of each brain section were determined for the size of the infarct focus using image analysis software. The overall brain area was calculated as follows: the brain area is left brain area × 2.
The volume of cerebral infarction was calculated by the following formula according to the method of determining the volume of the frustum:
[ S + S + √ (sS) ] × h/3 (where S: the area of slice 1; S: the area of slice 2; and h: the distance between slices).
The volumes of 3 sites between adjacent slices were determined and summed up, and the percentage of cerebral infarction volume was calculated:
cerebral infarct volume (%) -/whole brain volume × 100%.
And (3) statistical treatment:
the measurements are expressed as mean and standard error (mean + -SEM). Statistical analysis of the measurements was performed by one-way anova (dispersion analysis of unary configuration) using the table calculation software, and significant differences were considered to exist in the case of p < 0.05.
Neurological deficit scoring was performed 24h post reperfusion, according to Longa 5 scoring criteria.
0 minute: normal, no neurological impairment; 1 minute: the left anterior paw can not be fully extended, and mild neurological deficit is caused; and 2, dividing: when walking, the rat turns to the left side (paralyzed side), and the function of the nerve is damaged moderately; and 3, dividing: when walking, the rat body was inclined to the left (paralyzed side). Severe neurological impairment; and 4, dividing: spontaneous walking is not possible and consciousness is lost.
Test results
Example 3: the pharmaceutical composition tablet of the invention
Weighing 8g of phellodendrine and 16g of paeoniflorin, uniformly mixing, adding 160g of sodium carboxymethylcellulose and 50g of dextrin, sieving with a 16-mesh sieve to prepare wet granules, drying the wet granules at 50-60 ℃, grading the dry granules with a 20-mesh sieve, adding 2% of magnesium stearate, uniformly mixing, tabletting, and tabletting, wherein the tablet weight is 0.2 g/tablet.
Example 4: the invention relates to a dripping pill of a pharmaceutical composition
Weighing 4g of phellodendrine, 8g of paeoniflorin and EGCG4g, mixing with polyethylene glycol-400020 g and polyethylene glycol-60004 g, heating in water bath to melt, adding medicinal simethicone as coolant, dripping into pill at 70 deg.C, drying, and making into dripping pill of 25 mg/pill.
Example 5: the invention relates to a powder injection of a pharmaceutical composition
Weighing 4g of phellodendrine, 8g of paeoniflorin and EGCG4g, mixing with 10g of lactose, adding 100mL of water for injection, and heating at 80 ℃ to obtain a suspension for later use; weighing 20g of mannitol, adding 100mL of water for injection, slowly adding the suspension under stirring at 80 ℃, adding 0.05% of activated carbon for injection, stirring at 80 ℃ for 30min, filtering with a microporous membrane, metering the volume of the filtrate to 300mL, subpackaging, and freeze-drying to obtain the finished product, wherein the volume is 3 mL/piece.
The foregoing describes preferred embodiments of the present invention, but is not intended to limit the invention thereto. Modifications and variations of the embodiments disclosed herein may be made by those skilled in the art without departing from the scope and spirit of the invention.
Claims (7)
1. A pharmaceutical composition is characterized by comprising phellodendrine and paeoniflorin, wherein the weight ratio of phellodendrine to paeoniflorin is 1: 5.
2. A pharmaceutical composition is characterized by comprising phellodendrine, paeoniflorin and EGCG, wherein the weight ratio of the phellodendrine to the paeoniflorin to the EGCG is 1:5: 1.
3. The pharmaceutical composition of any one of claims 1-2, wherein the composition further optionally comprises a pharmaceutically acceptable carrier.
4. A method for preparing the composition as claimed in any one of claims 1-2, comprising the step of mixing phellodendrine, paeoniflorin and optionally EGCG and optionally pharmaceutically acceptable carrier uniformly according to the weight ratio to obtain the composition.
5. Use of the pharmaceutical composition according to any one of claims 1-2 for the preparation of a medicament for the prevention or treatment of brain tissue damage caused by local and global cerebral ischemia.
6. The use of the pharmaceutical composition according to claim 5 for the preparation of a medicament for the prevention or treatment of cerebral hemorrhage, cerebral infarction, cerebral thrombosis.
7. Use of the pharmaceutical composition according to claim 6 for the preparation of a medicament for the prevention or treatment of cerebral infarction.
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