Plant extract composition and application thereof

Abstract

The present application relates to the field of biomedicine technology, and more particularly to a plant extract composition and its application. The plant extract composition comprises a water-soluble tomato concentrate and an Ampelopsis grossularis extract. Compared to conventional technologies, the beneficial effects of the embodiments of the present application include: the plant extract composition comprising a water-soluble tomato concentrate and an Ampelopsis grossularis extract provided in the embodiments of the present application has a synergistic effect in improving blood flow and / or inhibiting platelet aggregation, and preventing and treating cerebral thrombosis. The combined use of the water-soluble tomato concentrate and Ampelopsis grossularis extract significantly improves the effect compared to the use of a single extract, greatly reduces the cost, and has excellent application prospects.

Description

Technical Field

[0001] The present application relates to the field of biomedicine technology, and in particular to a plant extract composition and its application. Background Art

[0002] Cerebral thrombosis is a common ischemic cerebrovascular disease with high mortality and disability rates. The main harm of cerebral thrombosis is vascular occlusion and blood flow interruption, resulting in ischemia, hypoxia, softening, and necrosis of brain tissue in the blood supply area, which in turn leads to neurological damage. Specific manifestations include hemiplegia, numbness of the limbs, motor impairment, slurred speech, dizziness, blurred vision, etc. Severe cases may cause coma, incontinence, and even death. In recent years, changes in dietary structure and an accelerated pace of life have led to an increasing incidence of cerebral thrombosis, and the affected population is tending to be younger, posing a significant threat to the health of the general population.

[0003] At present, prevention is the main approach for cerebral thrombosis and the neurological damage it causes, including primary prevention (for people at risk who have not developed cerebral thrombosis) and secondary prevention (for patients with cerebral thrombosis). Preventive measures are very necessary for both patients with cerebral thrombosis and people at high risk of cerebral thrombosis.

[0004] Recent studies have shown that abnormal platelet activation and aggregation play a key role in cerebral thrombosis, and antiplatelet therapy is recommended to reduce the risk of cerebral thrombosis. Currently, commonly used antiplatelet drugs in clinical practice include arachidonic acid cyclooxygenase inhibitors represented by aspirin, the purine receptor antagonist clopidogrel, and the platelet membrane glycoprotein IIb / IIIa receptor antagonist abciximab. Although the important role of aspirin in the prevention and treatment of cerebral thrombosis has been continuously revealed, its application has been limited due to "aspirin resistance" and adverse reactions such as bleeding. Other antiplatelet drugs are also mostly limited in clinical application due to the risk of bleeding. Based on the current clinical status, it is particularly important to provide new solutions for the prevention and treatment of cerebral thrombosis.

[0005] Provis Natural Products Co., Ltd. has developed a water-soluble tomato concentrate, dubbed a "natural aspirin," that has a very strong inhibitory effect on platelet aggregation and is free of adverse reactions such as gastrointestinal bleeding. It is expected to have promising applications in the prevention and treatment of cerebral thrombosis. However, since the water-soluble tomato concentrate is a patented raw material of Provis Natural Products Co., Ltd. and the extraction and purification yield is low, its high price significantly limits its application in the preparation of products for the prevention and treatment of cerebral thrombosis. Summary of the Invention

[0006] Based on this, one embodiment of the present application provides a plant extract composition, which can effectively improve blood flow, inhibit platelet aggregation, and prevent and treat cerebral thrombosis.

[0007] One embodiment of the present application provides a plant extract composition, which includes a water-soluble tomato concentrate and an Ampelopsis grossedentata extract.

[0008] In some embodiments of the present application, the weight ratio of the water-soluble tomato concentrate to the Ampelopsis grossedentata extract is 1:1-10.

[0009] In some embodiments of the present application, the weight ratio of the water-soluble tomato concentrate to the Ampelopsis grossedentata extract is 1:4.

[0010] Another embodiment of the present application provides a use of the plant extract composition in preparing a drug for preventing and treating cerebral thrombosis.

[0011] Yet another embodiment of the present application provides a drug for preventing and treating cerebral thrombosis, the drug comprising the plant extract composition as described above and pharmaceutically acceptable excipients.

[0012] In some embodiments of the present application, the dosage form of the drug is tablet, capsule or powder.

[0013] In some embodiments of the present application, the pharmaceutically acceptable excipients include one or more of a filler, a disintegrant, a binder, and a lubricant.

[0014] In some embodiments of the present application, the filler includes one or more of anhydrous sugar powder, lactose, sucrose, starch, microcrystalline cellulose and maltodextrin.

[0015] In some embodiments of the present application, the disintegrant includes one or more of sodium carboxymethyl starch, cross-linked polyvinylpyrrolidone, cross-linked sodium carboxymethyl cellulose, ethyl cellulose, methyl cellulose, sodium lauryl sulfate and polyvinyl pyrrolidone.

[0016] In some embodiments of the present application, the binder includes one or more of starch slurry, sodium carboxymethyl cellulose, hydroxypropyl cellulose, methyl cellulose, ethyl cellulose and hydroxypropyl methyl cellulose.

[0017] In some embodiments of the present application, the lubricant includes one or more of magnesium stearate, talc, polyethylene glycols and magnesium sulfate.

[0018] Yet another embodiment of the present application provides a use of the plant extract composition in preparing a health food for improving blood flow and / or inhibiting platelet aggregation.

[0019] Yet another embodiment of the present application provides a health food for improving blood flow and / or inhibiting platelet aggregation, wherein the health food comprises the plant extract composition and an excipient acceptable in food.

[0020] In some embodiments of the present application, the health food comprises tablets, capsules or powders.

[0021] In some embodiments of the present application, the acceptable excipients in the health food include one or more of fillers, excipients, disintegrants, lubricants and binders.

[0022] In some embodiments of the present application, the filler comprises one or more of anhydrous lactose, microcrystalline cellulose, and maltodextrin.

[0023] In some embodiments of the present application, the excipient includes maltodextrin.

[0024] In some embodiments of the present application, the disintegrant includes one or more of cross-linked sodium carboxymethyl cellulose, sodium carboxymethyl starch, and cross-linked polyvinylpyrrolidone.

[0025] In some embodiments of the present application, the lubricant includes one or more of magnesium stearate and silicon dioxide.

[0026] In some embodiments of the present application, the binder includes one or more of starch slurry, sodium carboxymethyl cellulose and hydroxypropyl cellulose.

[0027] Compared with conventional technologies, the advantages of the embodiments of the present application include:

[0028] The plant extract composition provided in the examples of the present application, including a water-soluble tomato concentrate and an Ampelopsis grossedentata extract, has a synergistic effect in improving blood flow, inhibiting platelet aggregation, and preventing and treating cerebral thrombosis. The combined use of the water-soluble tomato concentrate and the Ampelopsis grossedentata extract significantly improves the effect and greatly reduces the cost compared to the use of a single extract, and has very good application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] In order to more clearly illustrate the technical solutions in the embodiments of the present application and to more fully understand the present application and its beneficial effects, the following is a brief introduction to the drawings required for describing the embodiments. Obviously, the drawings described below are only some embodiments of the present application. Those skilled in the art can also derive other drawings based on these drawings without inventive effort.

[0030] Figure 1 The following are the staining results of cerebral thrombosis in zebrafish of normal control group, model control group and aspirin group;

[0031] Figure 2 This is the staining effect of zebrafish brain thrombosis in different dosage groups of sample 1;

[0032] Figure 3 This is the staining effect of zebrafish brain thrombosis in different dosage groups of sample 2;

[0033] Figure 4 This is a picture of the staining effect of zebrafish brain thrombosis in the sample 3, 4, and 5 treatment groups. DETAILED DESCRIPTION

[0034] Below in conjunction with accompanying drawing, embodiment and example, the application is described in further detail.It should be understood that these embodiment and example are only used to illustrate the application and are not used to limit the scope of the application, and the purpose of providing these embodiment and example is to make the understanding of the disclosure of the application more thorough and comprehensive.It should also be understood that the application can be implemented in many different forms and is not limited to the embodiment and example described herein. Those skilled in the art can make various changes or modifications without violating the connotation of the application, and the equivalent form obtained also falls within the protection scope of the application.In addition, in the description below, a large amount of specific details are given in order to provide a more complete understanding of the application.It should be understood that the application can be implemented without one or more of these details.

[0035] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art to which this application pertains. The terms used herein in the specification of this application are for the purpose of describing embodiments and examples only and are not intended to limit this application.

[0036] the term

[0037] Unless otherwise specified or incompatible herewith, the terms and phrases used herein shall have the following meanings:

[0038] The terms "and / or", "or / and", and "and / or" used herein include any one of two or more related listed items, and also include any and all combinations of the related listed items, wherein the arbitrary and all combinations include any combination of two related listed items, any more related listed items, or all related listed items. It should be noted that when at least three items are connected by at least two conjunctions selected from "and / or", "or / and", and "and / or", it should be understood that in this application, the technical solution undoubtedly includes technical solutions connected by "logical and" and also undoubtedly includes technical solutions connected by "logical or". For example, "A and / or B" includes three parallel solutions of A, B and A+B. For example, the technical solution of "A, and / or, B, and / or, C, and / or, D" includes any one of A, B, C, and D (that is, the technical solution of all being connected by "logical OR"), and also includes any and all combinations of A, B, C, and D, that is, the combination of any two or any three of A, B, C, and D, and also includes the four-item combination of A, B, C, and D (that is, the technical solution of all being connected by "logical AND").

[0039] In this application, "plurality", "multiple", "multiple times", "multiples", etc., unless otherwise specified, refer to a quantity greater than or equal to 2. For example, "one or more" means one or more than or equal to two.

[0040] As used herein, "combination thereof", "any combination thereof", "any combination thereof" and the like include all suitable combinations of any two or more of the listed items.

[0041] Herein, the “suitable” mentioned in “suitable combination”, “suitable method”, “any suitable method”, etc. shall be based on the ability to implement the technical solution of this application, solve the technical problems of this application, and achieve the expected technical effects of this application.

[0042] In this application, "further", "further", "particularly" and the like are used for descriptive purposes to indicate differences in content, but should not be understood as limiting the scope of protection of this application.

[0043] In this application, the terms "optionally," "optional," and "optional" mean optional or dispensable, i.e., they refer to either option being selected from two parallel options: "with" or "without." If a technical solution contains multiple "optional" clauses, each "optional" clause is independent unless otherwise specified and there are no contradictions or constraints.

[0044] In this application, the terms "first," "second," "third," "fourth," etc. in "the first aspect," "the second aspect," "the third aspect," "the fourth aspect," etc. are used for descriptive purposes only and should not be understood as indicating or implying relative importance or quantity, nor should they be understood as implicitly indicating the importance or quantity of the indicated technical features. Furthermore, "first," "second," "third," "fourth," etc. serve only as non-exhaustive enumeration and description, and should be understood not to constitute a closed-ended limitation on quantity.

[0045] In this application, the technical features described in an open manner include closed technical solutions composed of the listed features, and also include open technical solutions containing the listed features.

[0046] In this application, when referring to a numerical interval (i.e., a numerical range), unless otherwise specified, the optional numerical distribution is considered continuous within the above numerical interval and includes the two numerical endpoints of the numerical range (i.e., the minimum and maximum values), as well as each numerical value between the two numerical endpoints. Unless otherwise specified, when a numerical interval refers only to integers within the numerical interval, it includes the two endpoint integers of the numerical range, as well as each integer between the two endpoints. In this article, it is equivalent to directly listing each integer, such as t is an integer selected from 1 to 10, indicating that t is any integer selected from the group of integers consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10. In addition, when multiple ranges are provided to describe features or characteristics, these ranges can be merged. In other words, unless otherwise specified, the ranges disclosed herein should be understood to include any and all subranges included therein.

[0047] Unless otherwise specified, the temperature parameters in this application allow for both constant temperature treatment and temperature fluctuations within a certain temperature range. It should be understood that the constant temperature treatment allows for temperature fluctuations within the accuracy range of instrument control. Fluctuations within ranges such as ±5°C, ±4°C, ±3°C, ±2°C, and ±1°C are permitted.

[0048] In this application, % (w / w) and wt% both refer to weight percentage, % (v / v) refers to volume percentage, and % (w / v) refers to mass volume percentage.

[0049] All documents mentioned in this application are cited as references in this application, just as each document is cited as reference separately. Unless they conflict with the application purpose and / or technical solution of this application, the cited documents involved in this application are cited in their entirety and for all purposes. When cited documents are involved in this application, the definitions of relevant technical features, terms, nouns, phrases, etc. in the cited documents are also cited. When cited documents are involved in this application, the examples and preferred embodiments of the cited relevant technical features may also be incorporated into this application as references, but are limited to the ability to implement this application. It should be understood that when the cited content conflicts with the description in this application, the present application shall prevail or be adaptively amended according to the description in this application.

[0050] In the present application, a "composition" may be a combination of multiple substances, and further, may be used in combination, or may be a mixture of the combination.

[0051] In this application, "preventing and / or treating" have the same meaning and are used interchangeably. In this application, "preventing and / or treating" encompasses aspects of prevention, treatment, and adjuvant therapy. As used herein, "preventing and treating" refers to alleviating, slowing the progression of, attenuating, preventing, or maintaining an existing disease or condition. "Preventing and treating" also includes curing, preventing the development of, or alleviating to a certain degree, one or more symptoms of a disease or condition.

[0052] As used herein, "drug" includes any agent, compound, composition, or mixture that provides a pharmacological effect in vivo or in vitro, often with a beneficial effect. The scope of the pharmacological effect of a "drug" in vivo is not particularly limited and may be systemic or localized. The activity of the "drug" is not particularly limited and may be an active substance that interacts with other substances or an inert substance that does not interact.

[0053] As used herein, "pharmaceutically acceptable" refers to those ligands, materials, compositions, and / or dosage forms that are suitable for administration to a patient within the scope of sound medical judgment and commensurate with a reasonable benefit / risk ratio.

[0054] It should be understood that the term "medicament" includes a therapeutically effective amount of the plant extract composition described herein. As used herein, "therapeutically effective amount" refers to an amount of the active pharmaceutical ingredient that will elicit a biological or medical response in an individual to a disease, condition, and / or symptom, such as an amount of the plant extract composition described herein that produces a pharmacologically positive effect in an individual, including but not limited to reducing or inhibiting enzyme or protein activity, ameliorating symptoms, alleviating symptoms, slowing or delaying disease progression, or preventing disease.

[0055] The first aspect of the embodiments of the present application

[0056] The present embodiment provides a plant extract composition, which includes a water-soluble tomato concentrate and an Ampelopsis grossedentata extract.

[0057] The present application compounds a water-soluble tomato concentrate with an Ampelopsis grossedentata extract, thereby achieving a good effect in preventing and treating cerebral thrombosis while reducing the amount of the water-soluble tomato concentrate used and reducing costs.

[0058] In some examples, the weight ratio of the water-soluble tomato concentrate to the Ampelopsis grossedentata extract is 1:1-10 (e.g., 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10).

[0059] In some examples, the weight ratio of the water-soluble tomato concentrate to the Ampelopsis grossedentata extract is 1:4.

[0060] The second aspect of the embodiment of the present application

[0061] The present application provides an application of the plant extract composition in the preparation of a drug for preventing and treating cerebral thrombosis.

[0062] The third aspect of the embodiment of the present application

[0063] An embodiment of the present application provides a drug for preventing and treating cerebral thrombosis, which includes the plant extract composition as described above and pharmaceutically acceptable excipients.

[0064] The embodiments of the present application do not particularly limit the dosage form of the drug. Suitable pharmaceutically acceptable excipients can be selected according to clinical needs to prepare the corresponding dosage form, including but not limited to tablets, capsules or powders. Pharmaceutically acceptable excipients include but are not limited to fillers, disintegrants, binders and lubricants. The fillers include but are not limited to anhydrous sugar powder, lactose, sucrose, starch, microcrystalline cellulose and maltodextrin. The disintegrants include but are not limited to sodium carboxymethyl starch, cross-linked polyvinylpolypyrrolidone, cross-linked sodium carboxymethyl cellulose, ethyl cellulose, methyl cellulose, sodium lauryl sulfate and polyvinyl pyrrolidone. The binders include but are not limited to starch slurry, sodium carboxymethyl cellulose, hydroxypropyl cellulose, methyl cellulose, ethyl cellulose and hydroxypropyl methyl cellulose. The lubricants include but are not limited to magnesium stearate, talc, polyethylene glycols and magnesium sulfate.

[0065] The fourth aspect of the embodiments of the present application

[0066] The embodiments of the present application provide an application of the plant extract composition in the preparation of a health food for improving blood flow and / or inhibiting platelet aggregation.

[0067] In the present application, "health food" refers to food with health functions, which can regulate the physiological functions of animals (humans or other animals) and often provide beneficial effects. The scope of the beneficial effects of "health food" in the body is not particularly limited, and it can be a systemic effect or it can only produce effects locally. In the present application, "food" refers to a product that can be eaten directly. It should be understood that the food in this application may include the aforementioned plant extract composition, and is also allowed to include any other suitable edible substances. In some embodiments, other edible substances can be selected from substances allowed to be added in food management specifications, and further, substances prohibited from being added in food management specifications are not included. Unless otherwise specified, food management specifications refer to the current specifications at the time of production.

[0068] In this application, the "inhibition of platelet aggregation" in the health food of "inhibiting platelet aggregation" refers to inhibiting the abnormal increase in the platelet aggregation rate of the subject, which can lead to the occurrence of thrombosis and further induce the occurrence of diseases such as cerebral infarction, myocardial infarction, and angina pectoris.

[0069] In this application, the term "improving blood flow" in the context of a "health food for improving blood flow" refers to beneficially improving blood flow, i.e., "improving blood flow" brings beneficial effects to health. In certain embodiments, "improving blood flow" refers to increasing the blood flow velocity of a subject with abnormally slow blood flow.

[0070] The fifth aspect of the embodiment of the present application

[0071] The embodiments of the present application provide a health food for improving blood flow and / or inhibiting platelet aggregation, wherein the health food comprises the plant extract composition and an excipient acceptable in food.

[0072] In this application, "acceptable excipients in food" refer to edible raw materials, edible excipients or a combination of the two. Among them, "edible raw materials" refer to edible substances that can provide health functions alone, but are not plant extract compositions; "edible excipients" refer to edible substances that do not have the ability to provide health functions alone. It should be understood that "acceptable excipients in food" can be selected from substances allowed to be added in the health food management regulations, and further, do not include substances prohibited from being added in the health food management regulations. Unless otherwise specified, the health food management regulations refer to the current regulations at the time of production.

[0073] It should be understood that the health food contains a health-effective amount of the plant extract composition of the present application. The "health-effective amount" herein refers to an amount that can beneficially improve blood flow and / or inhibit platelet aggregation, and can exert the beneficial effects of improving blood flow and / or inhibiting platelet aggregation when consumed at a normal health food dosage.

[0074] The embodiments of the present application do not particularly limit the dosage form of the health food. Appropriate food-acceptable excipients can be selected according to health needs to prepare the corresponding dosage form, including but not limited to tablets, capsules or powders.

[0075] For example, if the health food is a tablet, acceptable excipients in the health food include, but are not limited to, fillers, excipients, disintegrants, lubricants, and binders. Fillers include, but are not limited to, anhydrous lactose, microcrystalline cellulose, and maltodextrin. Excipients include, but are not limited to, maltodextrin. Disintegrants include, but are not limited to, croscarmellose sodium, sodium carboxymethyl starch, and cross-linked polyvinylpyrrolidone. Lubricants include, but are not limited to, magnesium stearate and silicon dioxide. Binders include, but are not limited to, starch slurry, sodium carboxymethyl cellulose, and hydroxypropyl cellulose.

[0076] In this application, "subject" refers to a patient taking a drug or a consumer taking a health food.

[0077] In the present application, a "subject" is an animal, preferably a mammal, more preferably a human, and the subject includes but is not limited to consumers of health foods and patients with diseases, disorders and / or symptoms. The subject in the present invention is preferably a mammal. The term "mammal" mainly refers to warm-blooded vertebrate mammals, including but not limited to: cats, dogs, rabbits, bears, foxes, wolves, monkeys, deer, mice (such as rats, mice), pigs, cattle, sheep, horses, humans, etc., preferably primates, more preferably humans. In the present application, a "patient" refers to an animal, preferably a mammal, such as a human, and also such as a rat or mouse. Different subjects may result in different selection ranges of other ingredients in the aforementioned drugs or health foods.

[0078] The sixth aspect of the embodiments of the present application

[0079] An embodiment of the present application provides a method for preventing and treating cerebral thrombosis, comprising administering to a subject a therapeutically effective amount of the plant extract composition or the medicine described in the present application.

[0080] The seventh aspect of the embodiment of the present application

[0081] The embodiments of the present application provide a method for beneficially improving blood flow and / or inhibiting platelet aggregation, comprising administering to a subject a health-effective amount of the plant extract composition described in the present application, or administering to a subject a health-effective amount of a health food containing the plant extract composition described in the present application.

[0082] The embodiments of the present application will be described in detail below with reference to the examples. It should be understood that these examples are intended to illustrate the present application only and are not intended to limit the scope of the present application. The experimental methods for which specific conditions are not specified in the following examples are preferably referred to the guidance provided in the present application, and can also be based on the experimental manuals or conventional conditions in this area, or according to the conditions recommended by the manufacturer, or with reference to experimental methods known in the art.

[0083] In the following specific examples, the measured parameters of raw material components may have slight deviations within the range of weighing accuracy unless otherwise specified. For temperature and time parameters, acceptable deviations caused by instrument testing accuracy or operational accuracy are allowed.

[0084] Example 1: Preparation of composition

[0085] The preparation method of the water-soluble tomato concentrate is described in the prior patent "200980148421.6" of Provis Natural Products Co., Ltd. The water-soluble tomato concentrate is obtained by removing the fat-soluble components in tomatoes using a physical method and concentrating the water-soluble components.

[0086] The extract of Ampelopsis grossularis was purchased from the Qingping Chinese Medicine Market in Guangzhou. The preparation method of the Ampelopsis grossularis extract comprises: providing dried stems and / or leaves of Ampelopsis grossularis, crushing the dried stems and / or leaves, and extracting the dried stems and / or leaves with water and / or ethanol (preferably pure water as the extraction solvent, the ratio of the mass of the Ampelopsis grossularis dry product to the mass of water is 1:10-60, preferably 1:40) as a solvent, the extraction temperature is 30°C, the extraction time is 15min-3h (preferably 2h), and the extraction is performed 1-3 times (preferably 2 times). After collecting the extract, centrifuging (preferably pure water as the extraction solvent, the ratio of the mass of the Ampelopsis grossularis dry product to the mass of water is 1:10-60, preferably 1:40), filtering the supernatant, and concentrating the supernatant to a concentrate of 1 / 3-1 / 10 volume, and then spray drying the concentrate at 60-80°C to obtain the Ampelopsis grossularis extract.

[0087] Example 2: Verification of efficacy

[0088] 1. Testing materials

[0089] 1.1. Sample preparation information

[0090] Samples 1, 2, and 3 were all prepared with ultrapure water to a 20.0 mg / mL stock solution and stored at -20°C. Sample 1 was a water-soluble tomato concentrate, Sample 2 was an Ampelopsis radiata extract, Sample 3 was a water-soluble tomato concentrate + Ampelopsis radiata extract (1:1 ratio, w / w), Sample 4 was a water-soluble tomato concentrate + Ampelopsis radiata extract (ratio 1:10), and Sample 5 was a water-soluble tomato concentrate + Ampelopsis radiata extract (ratio 1:4). Positive control: Enteric-coated aspirin tablets (hereinafter referred to as aspirin), white tablets, batch number BJ54728, Bayer HealthCare Co., Ltd., stored in a cool, dry place. Prepared with DMSO to a 45.0 mg / mL stock solution and stored at -20°C.

[0091] Experimental Animals

[0092] Zebrafish were raised in fish farming water at 28°C (water quality: 200 mg of instant sea salt was added to 1 L of reverse osmosis water, conductivity was 450-550 μS / cm; pH was 6.5-8.5; hardness was 50-100 mg / L CaCO3). They were bred and provided by the Huante Fish Farming Center. The experimental animal use license number is SYXK(Zhejiang)2022-0004. The breeding and management of zebrafish met the requirements of the international AAALAC certification (certification number: 001458).

[0093] Zebrafish (albino) with a melanin allele mutation were bred naturally in pairs. Zebrafish aged 2 days post fertilization (dpf) were used to determine the maximum detectable concentration (MTC) of the samples for the prevention and treatment of cerebral thrombosis and to evaluate their efficacy.

[0094] 1.3. Instruments, consumables, and reagents

[0095] Dissecting microscope (SZX7, OLYMPUS, Japan); CCD camera (VertA1, Shanghai Tusen Vision Technology Co., Ltd., China); biological microscope (CX31, OLYMPUS, Japan); zebrafish heartbeat and blood analysis system (PB2084C, Viewpoint, USA); precision electronic balance (CP214, OHAUS, USA); 6-well plate (Zhejiang Bellanbo Biotechnology Co., Ltd., China).

[0096] Ponatinib (Batch No. 13771, MedChemExpress, USA); o-dianisidine (Batch No. MKBX3619V, Sigma, USA); dimethyl sulfoxide (DMSO, Batch No. BCCD8942, Sigma, Switzerland); methylcellulose (Batch No. C2004046, Shanghai Aladdin Biochemical Technology Co., Ltd., China).

[0097] 2. Detection Method

[0098] 2.1 Evaluation of efficacy (incidence) in preventing and treating cerebral thrombosis

[0099] 2-day-old zebrafish (albino) with a melanin allele mutation were randomly selected and plated in 6-well plates, with 30 zebrafish treated in each well (experimental group). Samples (concentrations shown in Table 1) were administered in water. A positive control group received aspirin at a concentration of 100 μg / mL. A normal control group and a model control group were also established, with a volume of 3 mL per well. Except for the normal control group, all other experimental groups received ponatinib in water to establish a zebrafish cerebral thrombosis model.

[0100] After treatment at 28°C for one day, the zebrafish were stained with o-dianisidine. After staining, each experimental group was observed under a dissecting microscope. The number of zebrafish with cerebral thrombosis was counted, and the incidence rate (%) of cerebral thrombosis in each experimental group was calculated. The statistical analysis results of this indicator were used to evaluate the efficacy (incidence) of the sample in preventing and treating cerebral thrombosis. Statistical analysis was performed using SPSS 26.0 software. A p < 0.05 indicated a statistically significant difference.

[0101] Table 1. Results of the experimental evaluation of the efficacy (incidence) of samples in preventing and treating cerebral thrombosis (n=30)

[0102]

[0103]

[0104] Compared with the model control group, *P<0.05, ***p<0.001.

[0105] Cerebral thrombosis inhibition rate (%) = (number of cerebral thrombi in the model control group - number of cerebral thrombi in the drug-treated group) / number of cerebral thrombi in the model group × 100%.

[0106] Under the experimental conditions, high doses of Samples 1 and 2 have the effect of preventing and treating cerebral thrombosis. After reducing the doses of Samples 1 and 2 and compounding them in different proportions, the three dose groups of Samples 3, 4, and 5 all have the effect of preventing and treating cerebral thrombosis, which is specifically manifested in reducing the incidence of cerebral thrombosis. See Table 1 for details. Figure 1 、 Figure 2 、 Figure 3 、 Figure 4. Because the zebrafish brain has typical vertebrate brain morphological characteristics and possesses coagulation factors and platelet receptors, cerebral thrombosis can be induced after taking a large amount of ponatinib. After red blood cell specific staining (red), the head thrombosis of zebrafish suffering from cerebral ischemia will be significantly increased than that of normal zebrafish. As shown in the figure, the normal control group did not add ponatinib, and the model control group and the sample group both took in equal amounts of ponatinib (ponatinib was taken into the zebrafish body by dissolving it in the fish farming water). After taking the sample for a period of time, the present embodiment performed red blood cell specific staining on the zebrafish as a whole and observed the thrombosis of the head. Among them, the model group had an increase in thrombosis, the aspirin group had a decrease in thrombosis, the sample 1 dosage group had a decrease in thrombosis as the dose increased, the sample 2 dosage group had more thrombosis, the high-dose group had a significant decrease in thrombosis, and the thrombosis of samples 3, 4, and 5 was significantly reduced.

[0107] The interaction between the components in the composition was further calculated according to the Jin Zhengjun Q value method, and the formula is as follows:

[0108] Q=E[a+b] / [Ea+(1-Ea)×Eb]; where:

[0109] E[a+b] is the inhibition rate when formula a and formula b are used together; Ea and Eb are the inhibition rates when formula a and formula b are used alone, respectively. When q < 0.85, it indicates that the two formulas have an antagonistic effect; when q > 1.15, it indicates that the two formulas have a synergistic effect.

[0110] The experimental results in Table 1 show that the inhibition rate of 25 μg / mL water-soluble tomato concentrate on cerebral thrombosis is 55.56%, the inhibition rate of 100 μg / mL Ampelopsis glauca extract on cerebral thrombosis is 3.70%, and the inhibition rate of 25 μg / mL water-soluble tomato concentrate and 100 μg / mL Ampelopsis glauca extract on cerebral thrombosis is 100%. It can be seen that the combination of the two is a q value of

[0111] 1.75, which is much greater than 1.15, so the combination of water-soluble tomato concentrate and Ampelopsis glauca extract shows obvious synergistic effect in inhibiting cerebral thrombosis.

[0112] 2.2 Evaluation of efficacy (blood flow) in preventing and treating cerebral thrombosis

[0113] 2-day-old zebrafish (albino) with a melanin allele mutation were randomly selected and plated in 6-well plates, with 30 zebrafish treated in each well (experimental group). Samples were administered in water (concentrations shown in Table 2). Aspirin at a concentration of 100 μg / mL was administered to the positive control group. A normal control group and a model control group were also established, with the volume per well being 3 mL. Except for the normal control group, all other experimental groups received ponatinib in water to establish a zebrafish cerebral thrombosis model. After one day of treatment at 28°C, 10 zebrafish were randomly selected from each experimental group and placed in a heartbeat blood analysis system to record blood flow videos. Blood flow velocity was analyzed, and statistical analysis of this indicator was used to evaluate the efficacy of the samples in preventing and treating cerebral thrombosis (blood flow). Statistical results are expressed as mean ± SE. Statistical analysis was performed using SPSS 26.0 software. P < 0.05 indicated statistical significance.

[0114] Table 2. Results of the experimental evaluation of the efficacy (blood flow) of the samples in preventing and treating cerebral thrombosis (n=10)

[0115]

[0116] Compared with the model control group, *p<0.05, **p<0.01.

[0117] The blood flow promotion rate was calculated as follows: blood flow promotion rate = blood flow velocity of the drug administration group - blood flow velocity of the model group / blood flow velocity of the model group × 100%.

[0118] The experimental results showed that the blood flow velocity of the model group was significantly reduced after administration of Ponatinib, which was statistically different from the normal group, indicating that the model was successful. The blood flow velocity of sample group 1 at 25, 100, and 125 μg / mL, sample group 2 at 2000 μg / mL, and samples 3, 4, and 5 were all significantly increased after administration.

[0119] 2.3. Evaluation of platelet aggregation rate

[0120] SD rats were anesthetized with 3% sodium pentobarbital (60 mg / kg), and blood was collected from the abdominal aorta. The blood was anticoagulated with 3.8% sodium citrate (1:9) and centrifuged at 200g for 10 minutes to prepare platelet-rich plasma (PRP). The remaining portion was centrifuged at 2000g for 10 minutes to prepare platelet-poor plasma (PPP). PPP was used to adjust the platelet count in PRP according to experimental needs. Counting method: Add 1.98mL of diluent (1% ammonium oxalate solution) to a clean test tube, accurately draw 20μL of PRP, place it in the platelet diluent, and immediately mix thoroughly. Take 10μL of the above-mentioned uniform platelet suspension, inject it into the counting chamber, let it stand for 10 minutes, and allow the platelets to sink. Count under a microscope.

[0121] Platelet aggregation rate measurement: First, connect the instrument to the computer, open the software, and then turn on the instrument. Preheat for 30 minutes. Note that the temperature displayed in the software indicates the connection between the computer and the instrument. Place a clean test cup in the incubation chamber and add a stir bar. Add a total of 300 μL of PRP and test sample / normal saline in a 9:1 ratio to the test cup. Zero the PPP and add equal amounts of test sample / normal saline of varying concentrations. Incubate the PRP and test sample for 4 minutes. To begin the measurement, insert the test cup containing the corresponding PPP into the colorimetric cell. Use the transmittance of the PPP to adjust the zero point. The system automatically adjusts the transmittance before aggregation to 100%. Press the corresponding channel button as prompted. Remove the PPP and then insert the PRP test cup containing the test sample of varying concentrations. The system records the transmittance at this time. Once the value stabilizes, press the corresponding channel button. Immediately add the corresponding amount of inducer (collagen). Immediately press the corresponding channel button, and the instrument begins measuring platelet aggregation over a 5-minute period (6 minutes for collagen induction). Repeat the experiment at least twice under the same experimental conditions, analyze the data statistically, and examine the stability of the experimental results. Determine the maximum platelet aggregation rate and calculate the inhibition rate of the test article on platelet aggregation according to the following formula.

[0122] Platelet aggregation inhibition rate = (maximum aggregation rate of control group - maximum aggregation rate of test group) / maximum aggregation rate of control group × 100%.

[0123] Table 3. Effects of samples on inhibiting collagen-induced platelet aggregation in vitro (Mean ± SD)

[0124]

[0125] *: Note: Compared with the blank control group, *P<0.05; **, P<0.01

[0126] The test results showed that the aspirin group, the low, medium and high dose groups of sample 1, the medium and high dose groups of sample 2, and the dose group of sample 5 had a significant inhibitory effect on platelet aggregation.

[0127] Under the experimental conditions of this example, Sample 1 showed efficacy in improving blood flow velocity, preventing and treating cerebral thrombosis, and inhibiting platelet aggregation; Sample 2 showed efficacy in improving blood flow velocity, preventing and treating cerebral thrombosis, and inhibiting platelet aggregation at high doses. Samples 3, 4, and 5, prepared by compounding low-dose Sample 1 and Sample 2, showed strong efficacy in improving blood flow velocity, preventing and treating cerebral thrombosis, and inhibiting platelet aggregation, and the two exhibited strong synergistic effects, as shown in Table 2.

[0128] The experiments in this example show that Sample 1, high-dose Sample 2, and Samples 3, 4, and 5 have the effects of improving blood flow velocity, preventing and treating cerebral thrombosis, and inhibiting platelet aggregation. Among them, the combination of water-soluble tomato concentrate and Snake extract can reduce the dosage of both, which has the effect of reducing dosage and increasing efficiency.

[0129] The various technical features of the above-mentioned implementation modes and examples can be combined in any appropriate manner. In order to make the description concise, not all possible combinations of the various technical features in the above-mentioned implementation modes and examples are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of the description in this specification.

Claims

1. A plant extract composition for preventing and treating cerebral thrombosis, characterized in that: The plant extract composition consists of a water-soluble tomato concentrate and an Ampelopsis grossedentata extract; The weight ratio of the water-soluble tomato concentrate to the Ampelopsis serrulate extract is 1:(1-10); The preparation method of the Ampelopsis grossedentata extract comprises: providing dried stems and / or leaves of Ampelopsis grossedentata, crushing the dried stems and / or leaves, and extracting with water and / or ethanol as a solvent at an extraction temperature of 30° C. for 15 minutes to 3 hours, and performing 1-3 extractions; collecting the extract, centrifuging, filtering, and concentrating the supernatant to a concentrate of 1 / 3 to 1 / 10 of its volume; and spray-drying the concentrate at 60-80° C. to obtain the Ampelopsis grossedentata extract.

2. The plant extract composition for preventing and treating cerebral thrombosis according to claim 1, characterized in that: The weight ratio of the water-soluble tomato concentrate to the Ampelopsis grossedentata extract is 1:

4.

3. Use of the plant extract composition for preventing and treating cerebral thrombosis according to any one of claims 1 to 2 in the preparation of a medicament for preventing and treating cerebral thrombosis.

4. A drug for preventing and treating cerebral thrombosis, characterized in that: The medicine comprises the plant extract composition according to any one of claims 1 to 2 and a pharmaceutically acceptable excipient.

5. The drug for preventing and treating cerebral thrombosis according to claim 4, characterized in that: The dosage form of the medicine is tablet, capsule or powder.

6. The drug for preventing and treating cerebral thrombosis according to claim 4 or 5, characterized in that: The pharmaceutically acceptable excipients include one or more of fillers, disintegrants, binders and lubricants.

7. The drug for preventing and treating cerebral thrombosis according to claim 6, characterized in that: The filler comprises one or more of anhydrous sugar powder, lactose, sucrose, starch, microcrystalline cellulose and maltodextrin.

8. The drug for preventing and treating cerebral thrombosis according to claim 6, characterized in that: The disintegrant includes one or more of sodium carboxymethyl starch, cross-linked polyvinylpyrrolidone, cross-linked sodium carboxymethyl cellulose, ethyl cellulose, methyl cellulose, sodium lauryl sulfate and polyvinyl pyrrolidone.

9. The drug for preventing and treating cerebral thrombosis according to claim 6, characterized in that: The binder includes one or more of starch slurry, sodium carboxymethyl cellulose, hydroxypropyl cellulose, methyl cellulose and ethyl cellulose.

10. The drug for preventing and treating cerebral thrombosis according to claim 6, characterized in that: The lubricant includes one or more of magnesium stearate, talc, polyethylene glycols and magnesium laurate sulfate.

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