Red apple and cherry plum composition and its application in anti-atherosclerosis

Abstract

The present invention relates to the field of food and medicine, and in particular to a red apple and cherry plum composition with anti-atherosclerotic efficacy, wherein the weight ratio of the red apple and cherry plum is 1:10-10:1. Mouse animal experiments show that the red apple and cherry plum composition has a synergistic effect in anti-atherosclerotic efficacy, and the efficacy of the composition in preventing or treating atherosclerosis is better than that of using red apple or cherry plum alone. The product forms of the red apple and cherry plum composition include juice, jam, fruit powder, oral liquid products, capsules, tablets, etc. The red apple and cherry plum composition also has a certain synergistic effect when used in combination with statins and other drugs.

Description

Technical Field

[0001] The invention relates to the field of food and medicine, and in particular to a red apple and cherry plum composition and application thereof in resisting arteriosclerosis. Background Art

[0002] Atherosclerosis (As) is an inflammatory disease that affects the arterial wall, characterized by the accumulation of lipids and inflammatory cells in the intima of large arteries. Atherosclerosis can cause atherosclerotic cardiovascular disease (ASCVD). Clinically diagnosed ASCVD diseases mainly include: acute coronary syndrome, history of myocardial infarction, stable or unstable angina, coronary artery revascularization (including interventional treatment and bypass surgery), other peripheral arterial disease or revascularization, atherosclerotic stroke or transient ischemic attack. Currently, ASCVD has become the leading cause of death in the world, seriously endangering health. In my country, the incidence of atherosclerosis has a clear upward trend, and it is more common in middle-aged and elderly people. Studies have shown that by 2020, 2 billion people in the world will have carotid atherosclerosis. A survey involving 85,000 people in my country showed that one-third of the elderly have atherosclerosis.

[0003] Clinical drugs for the treatment of atherosclerosis mainly include: (1) lipid-lowering drugs: statins, ezetimibe, PCSK9 inhibitors, etc.; (2) antiplatelet drugs: aspirin, clopidogrel, etc. The pathogenesis of AS is complex. A large number of studies have shown that low-density lipoprotein cholesterol (LDL-C) plays a key role in the occurrence and development of AS, and lipid-lowering drugs are the first choice. In addition, there are oxidative stress theory and inflammatory theory on the pathogenesis of atherosclerosis. There are some literature reports or clinical trials, such as colchicine and canakinumab related to the inflammatory theory, and probucol and idebenone related to oxidative stress. There are also some Chinese patent medicines for anti-atherosclerosis, such as compound Danshen dripping pills, Qishen Yiqi dripping pills, and Naoxintong capsules. Atherosclerosis requires a treatment plan under the guidance of a doctor, and long-term medication is required. Different drugs can have different degrees of side effects on the human body, such as liver and muscle damage, gastrointestinal reactions, etc., so a more natural and safe way of intervention is needed.

[0004] The red flesh apple (Malus neidzwetzkyana Dieck) is a plant of the genus Malus Mill in the family Rosaceae. The flesh of its fruit is red, and it has a very high potential utilization value in the research and processing of germplasm resources. There are many varieties of red flesh apples, and the fruit size and color vary greatly. my country, New Zealand, Japan, Switzerland and other countries have carried out breeding work on red flesh apples. Specifically, they include the Hongxun series in Xinjiang, China, Robert Crab in the United States, Ruby Sweet in Japan, and Red Love in Switzerland. Xinjiang's red flesh apples play an important role in the origin of red flesh apple plants.

[0005] Prunus cerasifera Ehrhart, commonly known as wild sour plum, is a plant of the genus Prunus in the Rosaceae family. Wild Prunus cerasifera is listed as a national Class II key protected species. It is a rare and endangered primitive wild fruit tree resource in the world. It is sparsely distributed naturally in the world. The Daxigou and Xiaoxigou in the Yili River Valley in Xinjiang is the world's largest concentrated distribution area of ​​wild Prunus cerasifera. Prunus cerasifera fruit has a long history of consumption in Xinjiang, my country. The Kazakh and Mongolian herdsmen living in the Daxigou of the Tianshan Mountains mainly eat beef and mutton, but few of them suffer from hyperlipidemia and hypertension. This is related to the herdsmen's long-term habit of picking wild Prunus cerasifera to make jam or drying the fruit for tea. Prunus cerasifera includes multiple varieties, which are mainly divided into black fruit, red fruit, purple fruit, purple-black fruit, yellow fruit, etc. according to color.

[0006] Red apples and cherry plums are rich in anthocyanins, polyphenols, flavonoids and other functional ingredients, and have high development value. However, current research is still limited, mainly focusing on chemical composition and antioxidants, and is still in the preliminary research stage. Summary of the invention

[0007] The inventors of the present application have found through a large number of experimental studies that the combination of red apple (or red apple extract) and cherry plum (or cherry plum extract) has a synergistic effect in resisting atherosclerosis.

[0008] To this end, in a first aspect of the present invention, the present invention provides a composition comprising:

[0009] A first component, wherein the first component is red flesh apple or a red flesh apple extract; and

[0010] The second component is cherry plum or cherry plum extract.

[0011] In some embodiments, the weight ratio of the first component to the second component is 1:10-10:1 (e.g., 1:9-10:1, 1:8-10:1, 1:7-10:1, 1:6-10:1, 1:5-10:1, 1:4-10:1, 1:3-10:1, 1:2-10:1, 1:1-10:1, 1:10-9:1, 1:9-9:1, 1:8-9:1, 1:7-9:1, 1:6-9:1, 1:5-9:1, 1:4-9:1, 1:3-9:1, 1:2-9:1, 1:1-9:1, 1:10-8:1, 1:9-8:1, 1:8-8:1, 1:7-8:1, 1:6-8:1, 1:5-8:1, 1 :4-8:1, 1:3-8:1, 1:2-8:1, 1:1-8:1, 1:10-7:1, 1:9-7:1, 1:8-7:1, 1:7-7:1, 1:6-7:1, 1:5-7:1, 1:4-7:1, 1:3-7:1, 1:2-7:1, 1:1-7:1, 1:10-6:1, 1:9- 6:1, 1:8-6:1, 1:7-6:1, 1:6-6:1, 1:5-6:1, 1:4-6:1, 1:3-6:1, 1:2-6:1, 1:1-6:1, 1:10-5:1, 1:9-5:1, 1:8-5:1, 1:7-5:1, 1:6-5:1, 1:5-5:1, 1:4-5:1, 1:3-5:1, 1:2-5:1, 1:1-5:1, 1:10-4:1, 1:9-4:1, 1:8-4:1, 1:7-4:1, 1:6-4:1, 1:5-4:1, 1:4-4:1, 1:3-4:1, 1:2-4:1, 1:1-4:1, 1:10-3:1, 1:9-3:1, 1:8 -3:1, 1:7-3:1, 1:6-3:1, 1:5-3:1, 1:4-3:1, 1:3-3:1, 1:2-3:1, 1:1-3:1, 1:10-2:1, 1:9-2:1, 1:8-2:1, 1:7-2:1, 1:6-2:1, 1:5-2:1, 1:4-2:1, 1:3-2:1 , 1:2-2:1, 1:1-2:1, 1:10-1:1, 1:9-1:1, 1:8-1:1, 1:7-1:1, 1:6-1:1, 1:5-1:1, 1:4-1:1, 1:3-1:1, 1:2-1:1, 1:10-5:1, 1:10-2:1, 1:10-1:1, 1:5-10:1 , 1:5-5:1, 1:5-2:1, 1:5-1:1, 1:2-10:1, 1:2-5:1, 1:2-2:1, 1:2-1:1, 1:1-10:1, 1:1-5:1, 1:1-2:1, such as 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, 1:1, 2:1,3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1).

[0012] In some embodiments, the red flesh apple or red flesh apple extract is obtained by one of the following methods: fresh fruit, fresh fruit pulping or juicing, immersion extraction, ultrasonic extraction, ultra-high pressure extraction, subcritical extraction, macroporous resin separation or other extraction methods.

[0013] In some embodiments, the cherry plum or cherry plum extract is obtained by one of the following methods: fresh fruit, fresh fruit pulping or juicing, immersion extraction, ultrasonic extraction, ultra-high pressure extraction, subcritical extraction, macroporous resin separation or other extraction methods.

[0014] In some embodiments, the composition is in the form of fresh fruit, juice, jam, fruit powder (such as freeze-dried powder), candy (such as compressed candy), oral liquid, capsule, tablet, dried fruit, preserved fruit, fruit wine or cake, etc. In other words, the product form of the composition can be fresh fruit, juice, jam, fruit powder (such as freeze-dried powder), candy (such as compressed candy), oral liquid, capsule, tablet, dried fruit, preserved fruit, fruit wine or cake, etc. That is to say, the composition can be made into fresh fruit, juice, jam, fruit powder (such as freeze-dried powder), candy (such as compressed candy), oral liquid, capsule, tablet, dried fruit, preserved fruit, fruit wine or cake, etc. Among them, juice, jam, fruit powder (such as freeze-dried powder), candy (such as compressed candy), oral liquid, capsule, tablet can be prepared by extracts obtained by leaching extraction, ultrasonic extraction, ultra-high pressure extraction, subcritical extraction, macroporous resin separation or other extraction methods.

[0015] In some embodiments, the composition is in the form of a medicine (such as Chinese medicine, Chinese patent medicine, natural medicine, etc.), a common food or a functional food or a health food. In other words, the composition can be made into a medicine (such as Chinese medicine, Chinese patent medicine, natural medicine, etc.), a common food or a functional food or a health food.

[0016] In some embodiments, the red flesh apple refers to the fruit of a plant of the genus Malus in the family Rosaceae having red flesh.

[0017] In some embodiments, the red flesh apple is selected from: Hongxun No. 1 (Hongxun No. 1), Hongxun No. 3 (Hongxun No. 3), Hongxun No. 4 (Hongxun No. 4), Hongxun No. 6 (Hongxun No. 6), Hongxun No. 8 (Hongxun No. 8), Red Love, etc.

[0018] In some embodiments, the cherry plum is selected from the group consisting of red cherry plum, black cherry plum, purple cherry plum, purple-black cherry plum, yellow cherry plum, and the like.

[0019] In a second aspect of the invention, the invention provides a combination product comprising:

[0020] A first product, wherein the first product is the aforementioned composition;

[0021] The second product is a drug for preventing and / or treating atherosclerotic diseases.

[0022] In some embodiments, the atherosclerotic disease is selected from atherosclerosis of the aorta and its main branches, coronary artery atherosclerosis, cerebral artery atherosclerosis, renal artery atherosclerosis, mesenteric artery atherosclerosis, limb artery atherosclerosis, etc.

[0023] In some embodiments, the drug for preventing and / or treating atherosclerotic diseases is selected from statins (such as rosuvastatin, atorvastatin, simvastatin), ezetimibe, PCSK9 inhibitors, aspirin, clopidogrel, colchicine, canakinumab, probucol, idebenone, etc.

[0024] In some embodiments, the weight ratio of the first product to the second product is 1000:0.001-1000:33.4 (such as 1000:0.002075, 1000:0.004175, 1000:0.0083, 1000:0.02075, 1000:0.04175, 1000:0.083, 1000:0.08325, 1000:0.167, 1000:0.20875, 1000:0. :0.3125, 1000:0.333, 1000:0.4175, 1000:0.62625, 1000:0.835, 1000:1.25, 1000:1.67, 1000:2.0875, 1000:2.505, 1000:2.875, 1000:4.175, 1000:8.35 or 1000:16.7, or such as 1000:0.002075-1000:16.7).

[0025] In some embodiments, when the second product is rosuvastatin, the weight ratio of the first product to the second product is 1000:0.01-1000:0.34 (such as 1000:0.02075, 1000:0.04175, 1000:0.083 or 1000:0.167, or such as 1000:0.02075-1000:0.167).

[0026] In some embodiments, when the second product is atorvastatin, the weight ratio of the first product to the second product is 1000:0.02-1000:0.67 (such as 1000:0.04175, 1000:0.08325, 1000:0.167 or 1000:0.333, or such as 1000:0.04175-1000:0.333).

[0027] In some embodiments, when the second product is aspirin, the weight ratio of the first product to the second product is 1000:0.10-1000:5.01 (such as 1000:0.20875, 1000:0.4175, 1000:0.62625, 1000:0.835, 1000:1.67 or 1000:2.505, or such as 1000:0.20875-1000:2.505).

[0028] In some embodiments, when the second product is clopidogrel, the weight ratio of the first product to the second product is 1000:0.10-1000:5.01 (such as 1000:0.20875, 1000:0.4175, 1000:0.62625, 1000:0.835, 1000:1.67 or 1000:2.505, or such as 1000:0.20875-1000:2.505).

[0029] In some embodiments, when the second product is colchicine, the weight ratio of the first product to the second product is 1000:0.001-1000:0.166 (such as 1000:0.002075, 1000:0.004175, 1000:0.0083, 1000:0.02075 or 1000:0.083, or such as 1000:0.002075-1000:0.083).

[0030] In some embodiments, when the second product is probucol, the weight ratio of the first product to the second product is 1000:1.0-1000:33.4 (such as 1000:2.0875, 1000:2.875, 1000:4.175, 1000:8.35 or 1000:16.7, or such as 1000:2.0875-1000:16.7).

[0031] In the third aspect of the present invention, the present invention provides the use of the aforementioned composition in the preparation of drugs (such as Chinese medicine, Chinese patent medicine, natural medicine, etc.), ordinary food or functional food or health food, and the drugs (such as Chinese medicine, Chinese patent medicine, natural medicine, etc.), ordinary food or functional food or health food are used to prevent and / or treat atherosclerotic diseases. The present invention also provides the use of the aforementioned combination product in the preparation of drugs (such as Chinese medicine, Chinese patent medicine, natural medicine, etc.), and the drugs (such as Chinese medicine, Chinese patent medicine, natural medicine, etc.) are used to prevent and / or treat atherosclerotic diseases.

[0032] In the present invention, ordinary food refers to finished products and raw materials for human consumption or drinking, as well as items that are traditionally both food and medicine, but does not include items for therapeutic purposes.

[0033] In the present invention, health food refers to food that claims to have specific health functions or is intended to supplement vitamins and minerals, that is, food that is suitable for consumption by specific groups of people, has the function of regulating body functions, is not intended to treat diseases, and does not cause any acute, subacute or chronic harm to the human body. Functional food is mainly a term in Europe, America and Japan for foods that can improve physical health or reduce illness, which is similar to the definition of health food in China.

[0034] In the present invention, Chinese medicine refers to medicines that are collected, processed, and prepared according to the theory of traditional Chinese medicine, explain the mechanism of action, and guide clinical application. Chinese medicines are mainly derived from natural medicines and their processed products, including herbal medicines, animal medicines, mineral medicines, and some chemical and biological medicines, with herbal medicines being the majority.

[0035] In the present invention, Chinese patent medicine is made of Chinese medicinal materials as raw materials, and is processed into Chinese medicine products of certain dosage forms according to prescribed prescriptions and preparation processes under the guidance of traditional Chinese medicine theory for the purpose of preventing and treating diseases. It is a type of commercialized Chinese medicine preparation approved by the national drug supervision and administration department.

[0036] In the present invention, natural medicine refers to animal medicine, plant medicine and mineral medicine that have been proven to have certain pharmacological activity by the modern medical system. Natural medicine is not equivalent to traditional Chinese medicine or Chinese herbal medicine. Natural medicine refers to natural medicinal substances and preparations used under the guidance of modern medical theory. Its sources include plants, animals and minerals, and generally do not include substances derived from genetically modified animals and plants, substances fermented by microorganisms or modified by chemicals, etc., which include a first product and a second product in independent packages, the first product is a combination product of red meat apples and cherry plums, and the second product is an anti-atherosclerotic drug (such as lipid-lowering drugs, antiplatelet drugs, anticoagulant drugs, anti-inflammatory and anti-oxidative stress drugs, etc.).

[0037] In the present invention, "individual" includes human or non-human animals. Exemplary human individuals include human individuals (referred to as patients) suffering from diseases (e.g., diseases described herein) or normal individuals. In the present invention, "non-human animals" include all vertebrates, such as non-mammals (e.g., fish, birds, amphibians, reptiles) and mammals, such as non-human primates, livestock and / or domesticated animals (e.g., mice, sheep, dogs, cats, cows, pigs, etc.).

[0038] In the present invention, "cherry plum extract" or "red flesh apple extract" refers to a product formed by obtaining and enriching one or more effective ingredients in cherry plum or red flesh apple through physical and / or chemical extraction and separation processes using cherry plum or red flesh apple as raw materials, which can be used as a raw material or auxiliary material for food (such as ordinary food or functional food or health food) or medicine. Exemplary extraction methods include but are not limited to: leaching extraction, ultrasonic extraction, ultra-high pressure extraction, subcritical extraction, macroporous resin separation, etc.

[0039] Exemplary methods for preparing red apple and cherry plum extracts are provided below.

[0040] Extraction

[0041] In certain embodiments, the red apple and cherry plum extracts of the present invention are obtained by leaching extraction (eg, water leaching extraction or alcohol leaching extraction).

[0042] In certain embodiments, the leaching extraction comprises the following steps:

[0043] Step 1: Remove the seeds or cores of red apples and cherry plums and then beat them to obtain pulp directly, or separate juice and pomace; in some embodiments, filter and separate the juice and pomace using gauze (e.g., 100-300 mesh gauze, e.g., 200 mesh gauze);

[0044] Step 2: Mix the pulp or pomace with an extraction medium (e.g., water or ethanol), add cellulase, extract at room temperature or under heating (e.g., 10-100° C., e.g., 50° C.) conditions (e.g., extract for 30-100 min, e.g., 60 min), and filter to obtain a pulp or pomace extract; in some embodiments, the pulp or pomace and the extraction medium are mixed in a weight ratio of 1:(2-10) (e.g., 1:5); in some embodiments, the ethanol is 20%-100% (e.g., 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100%) ethanol;

[0045] Step 3: Concentrating the pulp extract or the combined solution of the juice and pomace extract to obtain an extract of red apple and cherry plum; in some embodiments, the concentration is performed by rotary evaporation; in some embodiments, the rotary evaporation temperature is 30-60° C., for example, 50° C.;

[0046] Step 4: The extracted red apple and cherry plum extracts are mixed in different proportions (weight ratio) (1:10, 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1) to prepare a composition, or in step 1, the red apple and cherry plum raw materials are first mixed in the above-mentioned different proportions and then extracted.

[0047] In some embodiments, the red apple and cherry plum extracts of the present invention are obtained by water extraction. In some embodiments, the water extraction comprises the following steps:

[0048] Step 1: respectively remove the seeds or cores of red flesh apples and cherry plums and beat them to obtain pulp directly, or filter the juice with 100-300 mesh gauze to obtain red flesh apple and cherry plum juice, and separate the juice and pomace; in some embodiments, the mesh number of the gauze is 200 mesh;

[0049] Step 2: red apple and cherry plum pulp or pomace is mixed with water at a solid-liquid ratio of 1: (2-10), and cellulase is added, and the mixture is extracted at 10-100° C. for 30-100 min, and filtered to obtain a pulp or pomace extract; in some embodiments, the solid-liquid ratio is 1:5; in some embodiments, the extraction temperature is 50° C.; in some embodiments, the extraction time is 60 min; in some embodiments, the amount of cellulase added (based on the weight of the pomace or pulp) is 0.2%-1% (e.g., 0.5%);

[0050] Step 3: The red apple and cherry plum pulp extract or the combined solution of the juice and pomace extract is concentrated by rotary evaporation at 30-60° C. to obtain a red apple and cherry plum water extract; in some embodiments, the rotary evaporation temperature is 50° C.;

[0051] Step 4: The extracted red flesh apple and cherry plum water extracts are mixed in different proportions (weight ratio) (1:10, 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1) to prepare a composition, or the red flesh apple and cherry plum raw materials are first mixed in the above-mentioned different proportions in step 1 and then extracted.

[0052] In some embodiments, the red apple and cherry plum extracts of the present invention are obtained by alcohol extraction. In some embodiments, the alcohol extraction comprises the following steps:

[0053] Step 1: respectively remove the seeds or cores of red flesh apples and cherry plums and beat them to obtain pulp directly, or filter the juice with 100-300 mesh gauze to obtain red flesh apple and cherry plum juice, and separate the juice and pomace; in some embodiments, the mesh number of the gauze is 200 mesh;

[0054] Step 2: red apple and cherry plum pulp or pomace is mixed with 20%-100% (e.g., 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100%) of ethanol at a solid-liquid ratio of 1:(2-10), 0.2%-1% (based on the weight of the pomace or pulp) (e.g., 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9% or 1%) of cellulase is added, and the mixture is extracted at 10-100° C. for 30-100 min, and filtered to obtain a pulp or pomace extract; in some embodiments, the solid-liquid ratio is 1:5; in some embodiments, the concentration of the ethanol used is 40%; in some embodiments, the extraction temperature is 50° C.; in some embodiments, the extraction time is 60 min;

[0055] Step 3: The red apple and cherry plum pulp extract or the combined solution of the juice and pomace extract is concentrated by rotary evaporation at 30-60° C. The pulp extract is directly concentrated to obtain the red apple and cherry plum alcohol extract; in some embodiments, the rotary evaporation temperature is selected to be 50° C.; optionally, the extract is evaporated to remove residual ethanol therein;

[0056] Step 4: The extracted red apple and cherry plum alcohol extracts are mixed in different proportions (weight ratio) (1:10, 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1) to prepare a composition, or the red apple and cherry plum raw materials are first mixed in the above-mentioned different proportions in step 1 and then extracted.

[0057] Ultrasonic extraction

[0058] In certain embodiments, the red apple and cherry plum extracts of the present invention are obtained by ultrasonic extraction (eg, ultrasonic water extraction or ultrasonic alcohol extraction).

[0059] In certain embodiments, the ultrasonic extraction comprises the following steps:

[0060] Step 1: Remove the seeds or cores of red apples and cherry plums and then beat them to obtain pulp directly, or separate juice and pomace; in some embodiments, filter and separate the juice and pomace using gauze (e.g., 100-300 mesh gauze, e.g., 200 mesh gauze);

[0061] Step 2: Mix the pulp or pomace with an extraction medium (e.g., water or ethanol) and cellulase, extract under heating conditions (e.g., 30-50°C, e.g., 45°C) (e.g., extraction for 30-100 min, e.g., 60 min), then ultrasonicate under ultrasonic conditions (e.g., ultrasonic power of 100-500 W) (e.g., ultrasonicate for 20-60 min, e.g., ultrasonicate for 30 min), and filter to obtain a pulp or pomace extract; in some embodiments, the amount of cellulase added (based on the weight of the pomace or pulp) is 0.2%-1% (e.g., 0.5%); in some embodiments, the pulp or pomace is mixed with the extraction medium in a weight ratio of 1:(2-10) (e.g., 1:5); in some embodiments, the ethanol is 20%-100% (e.g., 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100%) ethanol;

[0062] Step 3: Concentrating the red apple and cherry plum pulp extract or the combined solution of the juice and pomace extract to obtain ultrasonic extracts of red apple and cherry plum; in some embodiments, the concentration is performed by rotary evaporation; in some embodiments, the rotary evaporation temperature is 30-60° C., for example, 50° C.;

[0063] Step 4: The extracted red apple and cherry plum ultrasonic extracts are mixed in different proportions (weight ratio) (1:10, 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1) to prepare a composition, or the red apple and cherry plum raw materials are first mixed in the above-mentioned different proportions in step 1 and then extracted.

[0064] In some embodiments, the red apple and cherry plum extracts of the present invention are obtained by ultrasonic water extraction. In some embodiments, the ultrasonic water extraction comprises the following steps:

[0065] Step 1: respectively remove the seeds or cores of red flesh apples and cherry plums and beat them to obtain pulp directly, or filter the juice with 100-300 mesh gauze to obtain red flesh apple and cherry plum juice, and separate the juice and pomace; in some embodiments, the mesh number of the gauze is 200 mesh;

[0066] Step 2: Mix red apple and cherry plum pulp or pomace with water at a solid-liquid ratio of 1: (2-10), add 0.2%-1% (based on the weight of the pomace or pulp) of cellulase, extract in a water bath at 30-50° C. for 30-100 min, ultrasonicate at 100-500 W for 20-60 min, and filter to obtain a pulp or pomace extract; in some embodiments, the solid-liquid ratio is 1:5; in some embodiments, the amount of cellulase added is 0.5%; in some embodiments, the water bath temperature is 45° C.; in some embodiments, the extraction time is 60 min; in some embodiments, the ultrasonic power is 500 W; in some embodiments, the ultrasonic time is 30 min;

[0067] Step 3: The combined solution of the red apple and cherry plum pulp or juice and the pomace extract is concentrated by rotary evaporation at 30-60° C. to obtain the red apple and cherry plum ultrasonic extraction water extract; in some embodiments, the rotary evaporation temperature is selected to be 50° C.;

[0068] Step 4: The extracted red apple and cherry plum ultrasonic water extracts are mixed in different proportions (weight ratio) (1:10, 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1) to prepare a composition, or the red apple and cherry plum raw materials are first mixed in the above-mentioned different proportions in step 1, and then extracted.

[0069] In some embodiments, the red apple and cherry plum extracts of the present invention are obtained by ultrasonic alcohol extraction. In some embodiments, the ultrasonic alcohol extraction comprises the following steps:

[0070] Step 1: respectively remove the seeds or cores of red flesh apples and cherry plums and beat them to obtain pulp directly, or filter the juice with 100-300 mesh gauze to obtain red flesh apple and cherry plum juice, and separate the juice and pomace; in some embodiments, the mesh number of the gauze is 200 mesh;

[0071] Step 2: Mix red apple and cherry plum pulp or pomace with 20%-100% ethanol at a solid-liquid ratio of 1: (2-10), add 0.2%-1% (based on the weight of the pomace or pulp) of cellulase, extract in a water bath at 30-50° C. for 30-100 min, ultrasonicate at 100-500 W for 20-60 min, and filter to obtain a pulp or pomace extract; in some embodiments, the solid-liquid ratio is 1:5; in some embodiments, the concentration of the ethanol used is 40%; in some embodiments, the amount of cellulase added is 0.5%; in some embodiments, the water bath temperature is 45° C.; in some embodiments, the extraction time is 60 min; in some embodiments, the ultrasonic power is 500 W; in some embodiments, the ultrasonic time is 30 min;

[0072] Step 3: The red apple and cherry plum pulp extract or the combined solution of the juice and pomace extract is concentrated by rotary evaporation at 30-60° C. to obtain the red apple and cherry plum ultrasonic extraction alcohol extract; in some embodiments, the rotary evaporation temperature is selected to be 50° C.; optionally, the extract is evaporated to remove residual ethanol therein;

[0073] Step 4: The extracted red apple and cherry plum ultrasonic alcohol extracts are mixed in different proportions (weight ratio) (1:10, 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1) to prepare a composition, or the red apple and cherry plum raw materials are first mixed in the above-mentioned different proportions in step 1 and then extracted.

[0074] Ultrahigh pressure extraction

[0075] In certain embodiments, the red apple and cherry plum extracts of the present invention are obtained by ultra-high pressure extraction (eg, ultra-high pressure water extraction or ultra-high pressure alcohol extraction). As used herein, "ultra-high pressure" refers to a pressure exceeding 100 MPa (equivalent to 1000 atmospheres).

[0076] In certain embodiments, the ultrahigh pressure extraction comprises the following steps:

[0077] Step 1: Remove the seeds or cores of red apples and cherry plums and then beat them to obtain pulp directly, or separate juice and pomace; in some embodiments, filter and separate the juice and pomace using gauze (e.g., 100-300 mesh gauze, e.g., 200 mesh gauze);

[0078] Step 2: Mix the pulp or pomace with an extraction medium (e.g., water or ethanol), treat under ultrahigh pressure (e.g., 200-600 MPa, e.g., 400 MPa) (e.g., for 3-10 min, e.g., for 5 min), and filter to obtain a pulp or pomace extract; in some embodiments, the pulp or pomace and the extraction medium are mixed in a weight ratio of 1:(2-10) (e.g., 1:5); in some embodiments, the ethanol is 50%-95% ethanol;

[0079] Step 3: Concentrating the combined solution of the red apple and cherry plum pulp or juice and the pomace extract to obtain an ultra-high pressure extract of red apple and cherry plum; in some embodiments, the concentration is performed by rotary evaporation; in some embodiments, the rotary evaporation temperature is 30-60° C., for example, 50° C.;

[0080] Step 4: The extracted red apple and cherry plum ultrahigh pressure extracts are mixed in different proportions (weight ratio) (1:10, 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1) to prepare a composition, or the red apple and cherry plum raw materials are first mixed in the above-mentioned different proportions in step 1 and then extracted.

[0081] In some embodiments, the red apple and cherry plum extracts of the present invention are obtained by ultra-high pressure water extraction. In some embodiments, the ultra-high pressure water extraction comprises the following steps:

[0082] Step 1: respectively remove the seeds or cores of red flesh apples and cherry plums and beat them to obtain pulp directly, or filter the juice with 100-300 mesh gauze to obtain red flesh apple and cherry plum juice, and separate the juice and pomace; in some embodiments, the mesh number of the gauze is 200 mesh;

[0083] Step 2: mixing red apple and cherry plum pulp or pomace with water at a solid-liquid ratio of 1:(2-10), treating at an ultra-high pressure of 200-600 MPa for 3-10 min, and filtering to obtain a pulp or pomace extract; in some embodiments, the solid-liquid ratio is 1:5; in some embodiments, the ultra-high pressure is 400 MPa; in some embodiments, the ultra-high pressure treatment time is 5 min;

[0084] Step 3: The red apple and cherry plum pulp extract or the combined solution of the juice and pomace extract is concentrated by rotary evaporation at 30-60° C. to obtain the red apple and cherry plum ultra-high pressure extraction water extract; in some embodiments, the rotary evaporation temperature is selected to be 50° C.;

[0085] Step 4: The ultra-high pressure water extracts of red apple and cherry plum obtained by extraction are mixed in different proportions (weight ratio) (1:10, 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1) to prepare a composition, or the red apple and cherry plum raw materials are first mixed in the above-mentioned different proportions in step 1 and then extracted.

[0086] In some embodiments, the red apple and cherry plum extracts of the present invention are obtained by ultra-high pressure alcohol extraction. In some embodiments, the ultra-high pressure alcohol extraction comprises the following steps:

[0087] Step 1: respectively remove the seeds or cores of red flesh apples and cherry plums and beat them to obtain pulp directly, or filter the juice with 100-300 mesh gauze to obtain red flesh apple and cherry plum juice, and separate the juice and pomace; in some embodiments, the mesh number of the gauze is 200 mesh;

[0088] Step 2: red apple and cherry plum pulp or pomace is mixed with 20%-100% (e.g., 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100%) ethanol at a solid-liquid ratio of 1:(2-10), treated at ultra-high pressure of 200-600 MPa for 3-10 min, and filtered to obtain a pulp or pomace extract; in some embodiments, the solid-liquid ratio is 1:5; in some embodiments, the ultra-high pressure is 400 MPa; in some embodiments, the ultra-high pressure treatment time is 5 min; in some embodiments, the ethanol is 40% ethanol;

[0089] Step 3: The red apple and cherry plum pulp extract or the combined solution of the juice and pomace extract is concentrated by rotary evaporation at 30-60° C. to obtain the red apple and cherry plum ultra-high pressure alcohol extract; in some embodiments, the rotary evaporation temperature is 50° C.; optionally, the extract is evaporated to remove residual ethanol therein;

[0090] Step 4: The extracted red apple and cherry plum ultrahigh pressure alcohol extracts are mixed in different proportions (weight ratio) (1:10, 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1) to prepare a composition, or the red apple and cherry plum raw materials are first mixed in the above-mentioned different proportions in step 1 and then extracted.

[0091] Subcritical extraction (also known as "subcritical extraction")

[0092] In certain embodiments, red apple and cherry plum extracts are obtained by subcritical extraction (eg, subcritical water extraction or subcritical alcohol extraction).

[0093] In certain embodiments, the subcritical extraction comprises the following steps:

[0094] Step 1: Remove the seeds or cores of red apples and cherry plums and then beat them to obtain pulp directly, or separate juice and pomace; in some embodiments, filter and separate the juice and pomace using gauze (e.g., 100-300 mesh gauze, e.g., 200 mesh gauze);

[0095] Step 2: Mix the pulp or pomace with an extraction medium (e.g., water or ethanol), perform subcritical extraction treatment (e.g., treatment for 10-50 min, e.g., 30 min), and filter to obtain a pulp or pomace extract; in some embodiments, the pulp or pomace and the extraction medium are mixed in a weight ratio of 1:(2-10) (e.g., 1:5); in some embodiments, the ethanol is 50%-95% ethanol; in some embodiments, the treatment temperature of the subcritical extraction is 100-150° C. (e.g., 110° C.);

[0096] Step 3: Concentrating the combined solution of the pulp or juice and the pomace extract to obtain a subcritical extract of red flesh apple and cherry plum; in some embodiments, the concentration is performed by rotary evaporation; in some embodiments, the rotary evaporation temperature is 30-60° C., for example, 50° C.;

[0097] Step 4: The extracted red flesh apple and cherry plum subcritical extracts are mixed in different proportions (weight ratio) (1:10, 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1) to prepare a composition, or in step 1, the red flesh apple and cherry plum raw materials are first mixed in the above-mentioned different proportions and then extracted.

[0098] In some embodiments, the red apple and cherry plum extracts of the present invention are obtained by subcritical water extraction. In some embodiments, the subcritical water extraction comprises the following steps:

[0099] Step 1: respectively remove the seeds or cores of red flesh apples and cherry plums and then beat them to obtain pulp directly, or filter the juice through 100-300 mesh gauze to obtain red flesh apple and cherry plum juice, and separate the juice and pomace; in some embodiments, the mesh number of the gauze is 200 mesh;

[0100] Step 2: mixing red apple and cherry plum pulp or pomace with water at a solid-liquid ratio of 1:(2-10), performing subcritical extraction for 10-50 min at a treatment temperature of 100-150° C., and filtering to obtain a pulp or pomace extract; in some embodiments, the solid-liquid ratio is 1:5; in some embodiments, the treatment temperature is 110° C.; in some embodiments, the treatment time is 30 min;

[0101] Step 3: The red apple and cherry plum pulp extract or the combined solution of the juice and pomace extract is concentrated by rotary evaporation at 30-60° C. to obtain a subcritical water extract of red apple and cherry plum; in some embodiments, the rotary evaporation temperature is 50° C.;

[0102] Step 4: The extracted red flesh apple and cherry plum subcritical water extracts are mixed in different proportions (weight ratio) (1:10, 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1) to prepare a composition, or the red flesh apple and cherry plum raw materials are first mixed in the above-mentioned different proportions in step 1, and then extracted.

[0103] In some embodiments, the red apple and cherry plum extracts of the present invention are obtained by subcritical alcohol extraction. In some embodiments, the subcritical alcohol extraction comprises the following steps:

[0104] Step 1: respectively remove the seeds or cores of red flesh apples and cherry plums and then beat them to obtain pulp directly, or filter the juice through 100-300 mesh gauze to obtain red flesh apple and cherry plum juice, and separate the juice and pomace; in some embodiments, the mesh number of the gauze is 200 mesh;

[0105] Step 2: mixing red apple and cherry plum pulp or pomace with 20%-100% (e.g., 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100%) ethanol at a solid-liquid ratio of 1:(2-10), performing subcritical extraction for 10-50 min at a treatment temperature of 100-150° C., and filtering to obtain a pulp or pomace extract; in some embodiments, the solid-liquid ratio is 1:5; in some embodiments, the treatment temperature is 110° C.; in some embodiments, the treatment time is 30 min;

[0106] Step 3: The red apple and cherry plum pulp extract or the combined solution of the juice and pomace extract is concentrated by rotary evaporation at 30-60° C. to obtain a subcritical alcohol extract of red apple and cherry plum; in some embodiments, the rotary evaporation temperature is 50° C.; optionally, the extract is evaporated to remove residual ethanol therein;

[0107] Step 4: The extracted red apple and cherry plum subcritical alcohol extracts are mixed in different proportions (weight ratio) (1:10, 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1) to prepare a composition, or the red apple and cherry plum raw materials are first mixed in the above-mentioned different proportions in step 1 and then extracted.

[0108] Macroporous resin separation

[0109] In certain embodiments, red apple and cherry plum extracts are separated using a macroporous resin.

[0110] In certain embodiments, the macroporous resin separation comprises the following steps:

[0111] Step 1: preparing red apple and cherry plum extracts by the above-mentioned different extraction methods (extraction, ultrasonic extraction, ultrahigh pressure extraction, subcritical extraction, etc.); or directly pulping the red apple and cherry plum to obtain pulp;

[0112] Step 2: Mix the extract or pulp of the red flesh apple and cherry plum with 95%-100% ethanol, the mixing ratio of the extract or pulp to the ethanol is 1: (2-10) (weight ratio) (for example, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9 or 1:10); the mixing time is 1-5 hours (for example, 3 hours), and then filtered to separate to obtain alcohol precipitate juice and precipitate, and the alcohol precipitate juice is subjected to rotary evaporation to remove ethanol, and the rotary evaporation temperature is 30-60°C (for example, 30°C, 40°C, 50°C or 60°C) to obtain alcohol precipitate clear juice; in some embodiments, the extract or red flesh apple and cherry plum extract can also be mixed with the extract or red flesh Pectinase is added to apple and cherry plum pulp for enzymatic clarification to obtain enzymatic clear juice, the enzymatic hydrolysis temperature is 20-60° C. (for example, 20° C., 30° C., 40° C., 50° C. or 60° C.), the enzymatic hydrolysis time is 30-240 min (for example, 30 min, 60 min, 90 min, 120 min, 150 min, 180 min, 210 min or 240 min), and the pectinase addition amount is 100-1000 ppm (for example, 100 ppm, 200 ppm, 300 ppm, 400 ppm, 500 ppm, 600 ppm, 700 ppm, 800 ppm, 900 ppm or 1000 ppm);

[0113] Step 3: using a macroporous adsorption resin to adsorb the alcohol precipitate juice or the enzymatic hydrolysis juice, and sequentially using water and 20%-100% ethanol (e.g., any of 20% ethanol, 40% ethanol, 60% ethanol, 80% ethanol, 100% ethanol) for gradient elution, and collecting the eluent of each gradient; in some embodiments, the elution rate is 1 to 5 BV / h (1 to 5 column volumes);

[0114] Step 4: The eluent is subjected to rotary evaporation to remove ethanol, and the rotary evaporation temperature is 30-60° C. (eg, 30° C., 40° C., 50° C. or 60° C.) to obtain samples separated by different macroporous resins.

[0115] Red apple and cherry plum can be extracted and prepared separately and then mixed in proportion to prepare a composition, or the red apple and cherry plum raw materials can be mixed in different proportions and then extracted to prepare the composition. As mentioned above, the product form of the composition of the present invention can be fresh fruit, juice, jam, fruit powder (such as freeze-dried powder), candy (such as compressed candy), oral liquid, capsule, tablet, dried fruit, preserved fruit, fruit wine or cake. Among them, juice, jam, fruit powder (such as freeze-dried powder), candy (such as compressed candy), oral liquid, capsule, tablet can be prepared by extracts obtained by leaching extraction, ultrasonic extraction, ultra-high pressure extraction, subcritical extraction, macroporous resin separation or other extraction methods. In addition, the product form of the composition of the present invention can be medicine, ordinary food or functional food or health food. In some embodiments, the composition of the present invention can be used as the main raw material in products such as juice, jam, fruit powder (such as freeze-dried powder), candy (such as compressed candy), oral liquid, capsule, tablet, dried fruit, preserved fruit, fruit wine or cake, for example, accounting for more than 30wt%, more than 40wt%, more than 50wt%, more than 60wt%, more than 70wt%, more than 80wt%, more than 90wt% or 100wt% of the total raw materials; optionally, the product may also contain other ingredients such as water, sweeteners, or other types of fruits or extracts thereof.

[0116] Exemplary methods for preparing red apple and cherry plum products (eg, fresh fruit, juice, jam, fruit powder, compressed candy, oral liquid, capsule) are provided below.

[0117] Fresh Fruit

[0118] In certain embodiments, fresh red apples and cherry plums can be obtained by a method comprising the following steps:

[0119] Step 1: Harvest red apples and cherry plums from one month before maturity to when they are fully ripe, and choose sunny days as the priority.

[0120] Step 2: Pick out and remove rotten fruits or fruits with poor color;

[0121] Step 3: Refrigerate and store after harvesting;

[0122] Step 4: Combine fresh red apples and cherry plums in different proportions (weight ratio) (1:10, 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1).

[0123] The fresh red-fleshed apples and cherry plums obtained can be eaten directly.

[0124] juice

[0125] In certain embodiments, red apple and cherry plum juice may be obtained by a method comprising the following steps:

[0126] Step 1: Remove the seeds or cores of red apples and cherry plums and blend them;

[0127] Step 2: filtering the slurry (for example, filtering with 100-300 mesh gauze) to obtain cloudy juice of red apple and cherry plum juice; in some embodiments, the mesh number of the gauze is 200 mesh;

[0128] Step 3: Mix red apple and cherry plum juice in different proportions (weight ratio) (1:10, 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1) to prepare a composition, or first mix the red apple and cherry plum raw materials in the above-mentioned different proportions in step 1, and then beat them to prepare the juice.

[0129] In certain embodiments, red apple and cherry plum juices can also be obtained by a method comprising the following steps:

[0130] Step 1: Remove the seeds or cores of red apples and cherry plums and blend them;

[0131] Step 2: filtering the slurry (for example, filtering with 100-300 mesh gauze), adding pectinase for enzymatic hydrolysis and filtering; obtaining red apple and cherry plum juice; in some embodiments, the mesh number of the gauze is 200 mesh;

[0132] Step 3: Mix red apple and cherry plum juice in different proportions (by weight) (1:10, 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1) to prepare a composition, or first mix the red apple and cherry plum raw materials in the above-mentioned different proportions in step 1, and then beat them to prepare the juice.

[0133] In certain embodiments, red apple and cherry plum juices can also be prepared using extracts obtained by the aforementioned leaching extraction, ultrasonic extraction, ultrahigh pressure extraction, subcritical extraction, macroporous resin separation or other extraction methods as raw materials.

[0134] jam

[0135] In certain embodiments, red apple and cherry plum jam can be obtained by a method comprising the following steps:

[0136] Step 1: Remove the seeds or cores of red apples and cherry plums and blend them;

[0137] Step 2: Boil and heat the fruit pulp to concentrate it into red apple and cherry plum jam;

[0138] Step 3: Mix red apple and cherry plum jam in different proportions (weight ratio) (1:10, 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1) to prepare a composition, or first mix the red apple and cherry plum raw materials in the above-mentioned different proportions in step 1, and then beat them to prepare jam.

[0139] In certain embodiments, red apple and cherry plum jams can also be prepared from extracts or juices obtained by the aforementioned leaching extraction, ultrasonic extraction, ultrahigh pressure extraction, subcritical extraction, macroporous resin separation or other extraction methods.

[0140] Fruit powder

[0141] In certain embodiments, red apple and cherry plum fruit powders may be obtained by a method comprising the following steps:

[0142] Step 1: Remove the seeds or cores of red apples and cherry plums and blend them.

[0143] Step 2: freeze-drying the fruit pulp to obtain fruit pulp freeze-dried powder;

[0144] Step 3: Mix the freeze-dried powder of red apple and cherry plum in different proportions (weight ratio) (1:10, 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1) to prepare a composition, or first mix the red apple and cherry plum raw materials in the above-mentioned different proportions in step 1, and then beat them to prepare freeze-dried powder.

[0145] In certain embodiments, red apple and cherry plum fruit powders may also be obtained by a method comprising the following steps:

[0146] Step 1: Remove the seeds or cores of red apples and cherry plums and blend them;

[0147] Step 2: filtering the slurry (for example, filtering with 100-300 mesh gauze) to obtain red apple and cherry plum juice; in some embodiments, the mesh number of the gauze is 200 mesh;

[0148] Step 3: freeze-drying the juice to prepare freeze-dried juice powder;

[0149] Step 4: Mix the freeze-dried powder of red apple and cherry plum in different proportions (weight ratio) (1:10, 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1) to prepare a composition, or first mix the red apple and cherry plum raw materials in the above-mentioned different proportions in step 1, and then beat them to prepare freeze-dried powder.

[0150] In certain embodiments, red apple and cherry plum fruit powders can also be prepared by belt drying according to the above steps to obtain fruit powders.

[0151] In certain embodiments, red apple and cherry plum fruit powders can also be prepared using extracts obtained by the aforementioned leaching extraction, ultrasonic extraction, ultrahigh pressure extraction, subcritical extraction, macroporous resin separation or other extraction methods as raw materials, and the fruit powders are prepared by freeze drying or belt drying.

[0152] Tablet Candy

[0153] In certain embodiments, red apple and cherry plum pressed candies may be obtained by a method comprising the following steps:

[0154] Step 1: Remove the seeds or cores of red apples and cherry plums and blend them;

[0155] Step 2: freeze-drying or belt-drying the fruit pulp into fruit powder;

[0156] Step 3: Compressing the fruit powder and auxiliary materials into tablet candies;

[0157] Step 4: Mix red apple and cherry plum pressed candies in different proportions (weight ratio) (1:10, 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1) to prepare a composition, or first mix the red apple and cherry plum raw materials in the aforementioned different proportions in step 1, and then beat them to prepare pressed candies.

[0158] In certain embodiments, red apple and cherry plum pressed candies can also be obtained by a method comprising the following steps:

[0159] Step 1: Remove the seeds or cores of red apples and cherry plums and blend them;

[0160] Step 2: filtering the slurry (for example, filtering with 100-300 mesh gauze) to obtain red apple juice; in some embodiments, the mesh number of the gauze is 200 mesh;

[0161] Step 3: freeze-dry or belt-dry the juice to make powder;

[0162] Step 4: Compress the fruit powder and auxiliary materials into tablet candies.

[0163] Step 5: Mix red apple and cherry plum pressed candies in different proportions (weight ratio) (1:10, 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1) to prepare a composition, or first mix the red apple and cherry plum raw materials in the aforementioned different proportions in step 1, and then beat them to prepare pressed candies.

[0164] In certain embodiments, red apple and cherry plum tablet candies can also be prepared using extracts obtained by the aforementioned leaching extraction, ultrasonic extraction, ultrahigh pressure extraction, subcritical extraction, macroporous resin separation or other extraction methods as raw materials.

[0165] Oral solution

[0166] In certain embodiments, the red apple and cherry plum oral solution can be obtained by a method comprising the following steps:

[0167] Step 1: preparing red apple and cherry plum extracts in different ratios (weight ratio) (1:10, 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1) (for example, prepared by fresh fruit pulping or juicing, leaching extraction, ultrasonic extraction, ultrahigh pressure extraction, subcritical extraction or macroporous resin separation, the steps are the same as above);

[0168] Step 2: sterilize the red flesh apple and cherry plum extracts to prepare the red flesh apple oral solution.

[0169] In certain embodiments, the red apple and cherry plum oral liquid can also be obtained by a method comprising the following steps:

[0170] Step 1: preparing red apple and cherry plum extracts in different ratios (weight ratio) (1:10, 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1) (for example, prepared by fresh fruit pulping or juicing, leaching extraction, ultrasonic extraction, ultrahigh pressure extraction, subcritical extraction or macroporous resin separation, the steps are the same as above);

[0171] Step 2: adding pectinase for enzymatic hydrolysis and filtering to prepare a clear juice extract composition;

[0172] Step 3: sterilize the red flesh apple and cherry plum juice extract to obtain the red flesh apple oral solution.

[0173] capsule

[0174] In certain embodiments, red apple and cherry plum capsules may be obtained by a method comprising the following steps:

[0175] Step 1: Remove the seeds or cores of red apples and cherry plums and blend them;

[0176] Step 2: freeze-drying or belt-drying the fruit pulp to obtain fruit pulp freeze-dried powder;

[0177] Step 3: mixing red apple and cherry plum fruit powders in different proportions (by weight) (1:10, 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1) to prepare a composite, or first mixing red apple and cherry plum raw materials in the above-mentioned different proportions in step 1, and then beating to prepare fruit powder;

[0178] Step 4: Fill the fruit powder into capsule shells to make capsules.

[0179] In certain embodiments, red apple and cherry plum capsules can also be obtained by a method comprising the following steps:

[0180] Step 1: Remove the seeds or cores of red apples and cherry plums and blend them;

[0181] Step 2: filtering the slurry (for example, filtering with 100-300 mesh gauze) to obtain red apple and cherry plum juice; in some embodiments, the mesh number of the gauze is 200 mesh;

[0182] Step 3: freeze-dry or belt-dry the juice to make powder;

[0183] Step 4: mixing red apple and cherry plum fruit powders in different proportions (weight ratio) (1:10, 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1) to prepare a composite, or first mixing red apple and cherry plum raw materials in the above-mentioned different proportions in step 1, and then beating to prepare fruit powder;

[0184] Step 5: Fill the fruit powder into capsule shells to make capsules.

[0185] In certain embodiments, red apple and cherry plum capsules can also be prepared using extracts obtained by the aforementioned leaching extraction, ultrasonic extraction, ultrahigh pressure extraction, subcritical extraction, macroporous resin separation or other extraction methods as raw materials.

[0186] Beneficial Effects

[0187] The present invention selects natural red apples and cherry plums, and the prepared red apple and cherry plum composition has a good anti-atherosclerotic effect. The effect of the two composites is better than that of using red apples or cherry plums alone, and has a synergistic effect. In addition, the effect of the composition further combined with a drug is better than that of the composition or the drug alone, and has a synergistic effect. DETAILED DESCRIPTION

[0188] The embodiments of the present invention will be described in detail below with reference to examples, but those skilled in the art will appreciate that the following examples are only used to illustrate the present invention and should not be construed as limiting the scope of the present invention.

[0189] Embodiment 1:

[0190] The red flesh apple and cherry plum products described in this embodiment refer to fresh red flesh apple and cherry plum fruits.

[0191] Red-fleshed apples (Hongxun No. 1) and cherry plums are harvested from one month before maturity to when they are fully mature. When picking fruits, sunny days are preferred. Rotten fruits or fruits with poor color are selected and removed, and they are refrigerated after picking. The obtained red-fleshed apples and cherry plums are combined in different proportions (weight ratio) (1:10, 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1), and the fresh fruits can be eaten directly.

[0192] Embodiment 2:

[0193] The red flesh apple and cherry plum products described in this embodiment refer to red flesh apple and cherry plum fruit juice.

[0194] After harvesting red apples (Hongxun No. 1) and cherry plums, the seeds or cores are removed and then pulped, and the juice is filtered through 200-mesh gauze to obtain red apple and cherry plum juice. The red apple and cherry plum juices are mixed in different proportions (weight ratio) (1:10, 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1) to prepare a composition, or the red apple and cherry plum raw materials are first mixed in the above different proportions and then pulped to prepare the juice.

[0195] Embodiment 3:

[0196] The red apple and cherry plum product described in this embodiment refers to red apple and cherry plum fruit juice.

[0197] After harvesting red apples (Hongxun No. 1) and cherry plums, the seeds or cores are removed and then pulped, and the juice is filtered with 200-mesh gauze to obtain cloudy red apple and cherry plum juices. Pectinase is added for enzymatic hydrolysis and filtration to obtain clear juice. The red apple and cherry plum clear juices are mixed in different proportions (weight ratio) (1:10, 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1) to prepare a composition, or the red apple and cherry plum raw materials are first mixed in the above-mentioned different proportions, and then pulped to prepare the juice.

[0198] Embodiment 4:

[0199] The red flesh apple and cherry plum product described in this embodiment refers to red flesh apple and cherry plum jam.

[0200] Red flesh apple (Hongxun No. 1) and cherry plum are pulped after being deseeded or cored, and boiled and heated to prepare thicker jam. Red flesh apple and cherry plum jam are mixed in different proportions (weight ratio) (1:10, 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1) to prepare a composite, or the red flesh apple and cherry plum raw materials are first mixed in the above different proportions and then pulped to prepare jam.

[0201] Embodiment 5:

[0202] The red flesh apple and cherry plum products described in this embodiment refer to red flesh apple and cherry plum fruit powder.

[0203] Red flesh apple (Hongxun No. 1) and cherry plum are pulped after deseeding or core removal, and freeze-dried to prepare fruit pulp freeze-dried powder. The freeze-dried powder of red flesh apple and cherry plum is mixed in different proportions (weight ratio) (1:10, 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1) to prepare a composite, or the red flesh apple and cherry plum raw materials are first mixed in the above different proportions, and then pulped to prepare fruit powder.

[0204] Embodiment 6:

[0205] The red flesh apple and cherry plum product described in this embodiment refers to red flesh apple and cherry plum pressed candy.

[0206] Red flesh apple (Hongxun No. 1) and cherry plum are pulped after being deseeded or cored, freeze-dried to obtain freeze-dried pulp powder, and the freeze-dried pulp powder and auxiliary materials are tableted to prepare tablet candies. Red flesh apple and cherry plum tablet candies are mixed in different proportions (weight ratio) (1:10, 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1) to prepare a composite, or the red flesh apple and cherry plum raw materials are first mixed in the above different proportions, and then pulped to prepare tablet candies.

[0207] Embodiment 7:

[0208] The red flesh apple and cherry plum product described in this embodiment refers to the oral liquid extracted from red flesh apple and cherry plum by water leaching.

[0209] (1) Red apples (Hongxun No. 1) and cherry plums are pulped after being deseeded or pitted to prepare red apple and cherry plum pulp.

[0210] (2) Water was added to the red apple and cherry plum pulp at a solid-liquid ratio of 1:5, and cellulase was added. The pulp was extracted at 50°C water for 60 minutes and filtered to obtain the pulp extract.

[0211] (3) The extract was concentrated by rotary evaporation at 50°C to obtain water extracts of red apple and cherry plum.

[0212] (4) The extracted red apple and cherry plum extracts were mixed in different proportions (weight ratio) (1:10, 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1) to prepare a composition.

[0213] (5) The combined extract of red apple and cherry plum is sterilized to prepare an oral solution.

[0214] Embodiment 8:

[0215] The red flesh apple and cherry plum products described in this embodiment refer to oral liquid extracted from red flesh apple (Hongxun No. 1) and cherry plum alcohol.

[0216] The treatment was carried out in accordance with steps (1) to (5) of Example 7, wherein the pulp was extracted with 20%, 40%, 60%, and 100% ethanol at a solid-liquid ratio of 1:5. The alcohol extract was evaporated to remove the ethanol.

[0217] Embodiment 9:

[0218] The red apple and cherry plum products described in this embodiment refer to the oral liquid extracted from red apple (Hongxun No. 3) and cherry plum. The preparation steps are the same as those in Example 7.

[0219] Embodiment 10:

[0220] The red flesh apple and cherry plum product described in this embodiment refers to the oral liquid extracted from red flesh apple (Hongxun No. 6) and cherry plum. The preparation steps are the same as those in Example 7.

[0221] Embodiment 11:

[0222] The red-fleshed apple and cherry plum product described in this embodiment refers to the ultrasonic water-extracted oral liquid of red-fleshed apple and cherry plum.

[0223] (1) Red apples (Hongxun No. 1) and cherry plums are pulped after being deseeded or pitted to prepare red apple and cherry plum pulp.

[0224] (2) Water was added to red apple and cherry plum pulp at a solid-liquid ratio of 1:5, 0.5% (based on the weight of the pulp) of cellulase was added, and the pulp was extracted in a 45°C water bath for 60 min, ultrasonicated at 500 W for 30 min, and filtered to obtain a pulp extract.

[0225] (3) The extract was concentrated by rotary evaporation at 50°C to obtain ultrasonic water extracts of red flesh apple and cherry plum.

[0226] (4) The extracted red apple and cherry plum extracts were mixed in different proportions (weight ratio) (1:10, 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1) to prepare a composition.

[0227] (5) The combined extract of red apple and cherry plum is sterilized to prepare an oral solution.

[0228] Embodiment 12:

[0229] The red apple and cherry plum products described in this embodiment refer to the ultrasonic water extraction oral liquid of red apple (Hongxun No. 6) and cherry plum. The preparation steps are the same as those in Example 10.

[0230] Embodiment 13:

[0231] The red flesh apple and cherry plum products described in this embodiment refer to the ultrasonic alcohol extraction oral liquid of red flesh apple (Hongxun No. 1) and cherry plum.

[0232] The treatment was carried out in accordance with steps (1) to (5) of Example 10, wherein the pulp was extracted with 20%, 40%, 60%, and 100% ethanol at a solid-liquid ratio of 1:5. The alcohol extract was evaporated to remove the ethanol.

[0233] Embodiment 14:

[0234] The red flesh apple and cherry plum products described in this embodiment refer to the oral liquid extracted from red flesh apple and cherry plum by ultra-high pressure water.

[0235] (1) Red apples (Hongxun No. 1) and cherry plums are pulped after being deseeded or pitted to prepare red apple and cherry plum pulp.

[0236] (2) Water was added to red apple and cherry plum pulp at a solid-liquid ratio of 1:5, and the pulp was treated at an ultrahigh pressure of 400 MPa for 5 min. The pulp extract was obtained by filtration.

[0237] (3) The extract was concentrated by rotary evaporation at 50°C to obtain ultra-high pressure water extracts of red apple and cherry plum.

[0238] (4) The extracted red apple and cherry plum extracts were mixed in different proportions (weight ratio) (1:10, 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1) to prepare a composition.

[0239] (5) The combined extract of red apple and cherry plum is sterilized to prepare an oral solution.

[0240] Embodiment 15:

[0241] The red flesh apple and cherry plum products described in this embodiment refer to the oral liquid extracted from red flesh apple (Hongxun No. 6) and cherry plum by ultra-high pressure water. The preparation steps are the same as those in Example 13.

[0242] Embodiment 16:

[0243] The red flesh apple and cherry plum products described in this embodiment refer to red flesh apple (Hongxun No. 1) and cherry plum ultra-high pressure alcohol extraction oral liquid.

[0244] The treatment was carried out in accordance with steps (1) to (5) of Example 13, wherein 50% of the pulp was added at a solid-liquid ratio of 1:5. The alcohol extract was evaporated to dry the ethanol.

[0245] Embodiment 17:

[0246] The red flesh apple and cherry plum products described in this embodiment refer to the subcritical water extracted oral liquid of red flesh apple and cherry plum.

[0247] (1) Red apples (Hongxun No. 1) and cherry plums are pulped after being deseeded or pitted to prepare red apple and cherry plum pulp.

[0248] (2) Water was added to the red apple and cherry plum pulp at a solid-liquid ratio of 1:5, and subcritical extraction was performed for 30 minutes at a processing temperature of 110°C. The pulp extract was obtained by filtration.

[0249] (3) The extract was concentrated by rotary evaporation at 50°C to obtain subcritical water extracts of red apple and cherry plum.

[0250] (4) The extracted red apple and cherry plum extracts were mixed in different proportions (weight ratio) (1:10, 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1) to prepare a composition.

[0251] (5) The combined extract of red apple and cherry plum is sterilized to prepare an oral solution.

[0252] Embodiment 18:

[0253] The red flesh apple and cherry plum product described in this embodiment refers to red flesh apple and cherry plum jam.

[0254] Referring to Example 7, first prepare the oral liquid of water extraction of red flesh apple (Hongxun No. 1) and cherry plum, then boil and heat to make thicker jam, with or without adding auxiliary materials. Mix red flesh apple and cherry plum jam in different proportions (weight ratio) (1:10, 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1) to prepare a composition, or first mix the red flesh apple and cherry plum raw materials in the above different proportions, and then prepare the jam.

[0255] Embodiment 19:

[0256] The red flesh apple and cherry plum products described in this embodiment refer to red flesh apple and cherry plum fruit powder.

[0257] Referring to Example 7, first prepare the oral liquid extracted from red flesh apple (Hongxun No. 1) and cherry plum, and then freeze-dry to prepare freeze-dried powder, with or without adding auxiliary materials. The freeze-dried powder of red flesh apple and cherry plum is mixed in different proportions (weight ratio) (1:10, 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1) to prepare a composition, or first mix the red flesh apple and cherry plum raw materials in the above different proportions, and then prepare the fruit powder.

[0258] Embodiment 20:

[0259] The red flesh apple and cherry plum product described in this embodiment refers to red flesh apple and cherry plum jam.

[0260] Referring to Example 11, first prepare red apple (Hongxun No. 1) and cherry plum ultrasonic extraction oral liquid, then boil and heat to make thicker jam, add or not add auxiliary materials. Red apple and cherry plum jam are mixed in different proportions (weight ratio) (1:10, 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1) to prepare a composite, or first mix red apple and cherry plum raw materials in the above different proportions, and then prepare jam.

[0261] Embodiment 21:

[0262] The red flesh apple and cherry plum products described in this embodiment refer to red flesh apple and cherry plum fruit powder.

[0263] Referring to Example 11, first prepare red apple (Hongxun No. 1) and cherry plum ultrasonic extraction oral liquid, then belt dry to obtain fruit powder, with or without adding auxiliary materials. Red apple and cherry plum fruit powder are mixed in different proportions (weight ratio) (1:10, 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1) to prepare a composite, or first mix red apple and cherry plum raw materials in the above different proportions, and then prepare fruit powder.

[0264] Embodiment 22:

[0265] The red flesh apple and cherry plum product described in this embodiment refers to the red flesh apple and cherry plum pure and clear juice product.

[0266] Referring to Example 7, a water-extracted concentrated solution of red flesh apple (Hongxun No. 1) and cherry plum was first prepared, and mixed with 95% ethanol for 3 hours. The weight ratio of the concentrated solution to 95% ethanol was 1:2. The concentrated solution was then filtered and separated to obtain an alcohol-precipitated clear juice. The ethanol was removed by rotary evaporation to obtain an alcohol-precipitated clear juice product of a red flesh apple and cherry plum composition.

[0267] Embodiment 23:

[0268] The red flesh apple and cherry plum products described in this embodiment refer to the red flesh apple and cherry plum macroporous resin separation products.

[0269] Referring to Example 22, first prepare red apple (Hongxun No. 1) and cherry plum alcohol precipitate juice products, use macroporous adsorption resin to adsorb the above clarified extract, and use water, 20%, 40%, 60%, and 100% ethanol for gradient elution in sequence, and the elution rate is 3BV / h (3 column volumes), and the eluate of each gradient is collected separately; the eluate is rotary evaporated to remove ethanol, and the rotary evaporation temperature is 30-60°C to obtain samples separated by different macroporous resins.

[0270] Evaluation of the anti-atherosclerotic effect of the above-mentioned embodiment products:

[0271] Experimental Example 1: Evaluation of the preventive effect of different proportions of red apple and cherry plum combinations on atherosclerosis in mice

[0272] (1) Test method

[0273] The wild-type C57 mice were used as the normal control group and rosuvastatin was used as the positive drug to investigate the efficacy of the mixed composition sample in preventing atherosclerosis in the high-fat diet-induced ApoE- / - mouse atherosclerosis model. The atherosclerosis model was established after 12 weeks of modeling (i.e., the model group), and the control group was fed with ordinary feed for 12 weeks (i.e., the normal group). The sample administration group was given the test sample at the same time as the modeling, and the normal group and the model group were given the same dose of distilled water. After 12 weeks of continuous administration, the formation of atherosclerotic plaques in the mouse aorta was observed, and the plaque area was dissected and the percentage of plaques was statistically analyzed.

[0274] (2) Test results

[0275] The test results are shown in Table 1. In the atherosclerosis prevention model, red apple and cherry plum were extracted by the water extraction method of Example 7 to obtain red apple water extract and cherry plum water extract. The red apple extract and cherry plum extract had a certain effect when used alone at a dose of 40 g / kg. When used in combination within the range of 1:10 to 10:1 (the combination dose was 40 g / kg), they had a more significant effect compared with the model. Compared with the model group, the formation of plaques in the arterial wall can be significantly reduced, and it has the effect of preventing atherosclerosis.

[0276] The q value was calculated according to the Jin's formula reported in the literature: q = E (A + B) / (EA + EB - EA EB)

[0277] Among them: E(A+B) is the inhibition rate of the two drugs in combination, EA and EB are the inhibition rates of each drug alone, q<1 indicates that the two drugs have an antagonistic effect when used together, and q≥1 indicates that the two drugs have a synergistic effect when used together.

[0278] As can be seen from Table 1, when red apples and cherry plums are used in combination, the q value is greater than or equal to 1, and the effect is significantly better than when used alone, and has a synergistic effect on the prevention of atherosclerosis.

[0279] Table 1 Preventive effect of different proportions of red apple and cherry plum combination on atherosclerosis in mice

[0280] Group Plaque area% Compared with the model, the inhibition rate is % q Model 33.02％ / / Positive drug (rosuvastatin) 1.67mg / kg 20.93％** 36.61％ / Red flesh apple 40g / kg 23.21％* 29.71％ / Cherry plum 40g / kg 25.25％* 23.53％ / Red apple: cherry plum 5:1 40g / kg 17.59％** 46.73％ 1.01 Red apple: cherry plum 1:1 40g / kg 15.64％** 52.63％ 1.14 Red apple: cherry plum 1:5 40g / kg 16.92％** 48.76％ 1.05

[0281] The significance in the above table is compared with the model group; *p<0.05, **p<0.01, ***p<0.001

[0282] Experimental Example 2: Evaluation of the therapeutic effect of different proportions of red apple and cherry plum combination on atherosclerosis in mice

[0283] (1) Test method

[0284] Wild-type C57 mice were used as the normal control group, and rosuvastatin was used as the positive drug to investigate the anti-atherosclerotic efficacy (i.e., therapeutic effect) of the mixed composition sample in the high-fat diet-induced ApoE- / - mouse atherosclerosis model. The atherosclerosis model was established 17 weeks after modeling, and the control group was fed with ordinary feed for 17 weeks. After 17 weeks of modeling, the sample administration group was continuously administered for 4 weeks to observe the formation of atherosclerotic plaques in the aorta of mice, and the plaque area was dissected and the percentage of plaques was statistically analyzed.

[0285] (2) Test results

[0286] The test results are shown in Table 2. In the atherosclerosis treatment model, red apple and cherry plum were extracted by the water extraction method of Example 7 to obtain red apple water extract and cherry plum water extract. The red apple extract and cherry plum extract had a certain effect when used alone at a dose of 40 g / kg. When used in combination within the range of 1:10 to 10:1 (the combination dose was 40 g / kg), they had a more significant effect compared with the model, and could significantly reduce the plaque area in the arterial wall, and had the effect of treating atherosclerosis.

[0287] The q value was calculated by referring to King's formula in Experimental Example 1.

[0288] As can be seen from Table 2, when red apple and cherry plum are used in combination, the q value is greater than or equal to 1, and the effect is significantly better than when used alone, and has a synergistic effect on the treatment of atherosclerosis.

[0289] Table 2 The therapeutic effect of different proportions of red apple and cherry plum combination on atherosclerosis in mice

[0290] Group Plaque area% Compared with the model, the inhibition rate is % q Model 42.57％ / / Positive drug (rosuvastatin) 1.67mg / kg 26.10％* 38.69％ / Red flesh apple 40g / kg 31.05％* 27.06％ / Cherry plum 40g / kg 30.23％* 28.99％ / Red apple: cherry plum 5:1 40g / kg 21.26％** 50.06％ 1.04 Red apple: cherry plum 2:1 40g / kg 20.98％** 50.72％ 1.05 Red apple: cherry plum 1:2 40g / kg 20.88％** 50.95％ 1.06 Red apple: cherry plum 1:5 40g / kg 20.84％** 51.05％ 1.06

[0291] The significance in the above table is compared with the model group; *p<0.05, **p<0.01, ***p<0.001

[0292] Experimental Example 3: Evaluation of the preventive effect of different doses of red apple and cherry plum combination on atherosclerosis in mice

[0293] (1) Test method

[0294] The wild-type C57 mice were used as the normal control group and rosuvastatin was used as the positive drug to investigate the efficacy of the mixed composition sample in preventing atherosclerosis in the high-fat diet-induced ApoE- / - mouse atherosclerosis model. The atherosclerosis model was established after 12 weeks of modeling (i.e., the model group), and the control group was fed with ordinary feed for 12 weeks (i.e., the normal group). The sample administration group was given the test sample at the same time as the modeling, and the normal group and the model group were given the same dose of distilled water. After 12 weeks of continuous administration, the formation of atherosclerotic plaques in the mouse aorta was observed, and the plaque area was dissected and the percentage of plaques was statistically analyzed.

[0295] (2) Test results

[0296] The test results are shown in Table 3. In the atherosclerosis prevention model, the effects of different doses of red apple and cherry plum compositions were compared. The water extraction method of Example 7 was used to extract the red apple water extract and the cherry plum water extract. The red apple extract and the cherry plum extract were weak when used alone at a dose of 20g / kg and 10g / kg, respectively. When used in combination (the composition dose was 20g / kg or 10g / kg, respectively), they had a more significant effect compared to the model, and could significantly reduce the formation of plaques in the arterial wall, and had the effect of preventing atherosclerosis.

[0297] The q value was calculated by referring to King's formula in Experimental Example 1.

[0298] As can be seen from Table 3, the effect of combined use is better than that of single use, and the q value is greater than or equal to 1, which shows a synergistic effect. Therefore, it can be seen that different doses of the combination of red apple and cherry plum can produce a synergistic effect in the prevention of atherosclerosis in mice.

[0299] Table 3 Effect of different doses of red apple and cherry plum combination on preventing atherosclerosis in mice

[0300] Group Plaque area% Compared with the model, the inhibition rate is % q Model 35.77％ / / Positive drug (rosuvastatin) 1.67mg / kg 24.82％ 30.61％ / Red flesh apple 20g / kg 27.37％ 23.48％ / Cherry plum 20g / kg 28.88％ 19.26％ / Red apple: cherry plum 1:2 20g / kg 20.95％ 41.43％ 1.08 Red apple: cherry plum 2:1 20g / kg 22.03％ 38.41％ 1.01 Red flesh apple 10g / kg 31.92％ 10.76％ / Cherry plum 10g / kg 31.40％ 12.22％ / Red apple: cherry plum 1:2 10g / kg 26.85％ 24.94％ 1.15 Red apple: cherry plum 2:1 10g / kg 25.63％ 28.35％ 1.31

[0301] The significance in the above table is compared with the model group; *p<0.05, **p<0.01, ***p<0.001

[0302] Experimental Example 4: Evaluation of the therapeutic effect of different doses of red apple and cherry plum combination on atherosclerosis in mice

[0303] (1) Test method

[0304] Wild-type C57 mice were used as the normal control group, and rosuvastatin was used as the positive drug to investigate the anti-atherosclerotic efficacy (i.e., therapeutic effect) of the mixed composition sample in the high-fat diet-induced ApoE- / - mouse atherosclerosis model. The atherosclerosis model was established 17 weeks after modeling, and the control group was fed with ordinary feed for 17 weeks. After 17 weeks of modeling, the sample administration group was continuously administered for 4 weeks to observe the formation of atherosclerotic plaques in the aorta of mice, and the plaque area was dissected and the percentage of plaques was statistically analyzed.

[0305] (2) Test results

[0306] The test results are shown in Table 4 below. In the atherosclerosis treatment model, the effects of different doses of red apple and cherry plum compositions were compared. Red apple and cherry plum were extracted by the water extraction method of Example 7 to obtain red apple water extract and cherry plum water extract. The red apple extract and cherry plum extract were weak when used alone at a dose of 20 g / kg and 10 g / kg, respectively. When used in combination (the dose of the composition was 20 g / kg or 10 g / kg, respectively), the effect was more significant than the model, and the plaques in the arterial wall were significantly reduced.

[0307] The q value was calculated by referring to King's formula in Experimental Example 1.

[0308] It can be seen from Table 4 that the combined use effect is better than the effect when used alone, and the q value is greater than or equal to 1, which shows a synergistic effect. Therefore, it can be seen that different doses of the combination of red apple and cherry plum can produce a synergistic effect in the treatment of atherosclerosis in mice.

[0309] Table 4 The therapeutic effect of different doses of red apple and cherry plum combination on atherosclerosis in mice

[0310]

[0311]

[0312] The significance in the above table is compared with the model group; *p<0.05, **p<0.01, ***p<0.001

[0313] Experimental Example 5: Evaluation of the therapeutic effect of the combination of the composition and statins on atherosclerosis in mice

[0314] (1) Test method

[0315] The wild-type C57 mice were used as the normal control group, and rosuvastatin and atorvastatin were used as positive drugs to investigate the anti-atherosclerotic efficacy (i.e., therapeutic effect) of the mixed composition sample in the high-fat diet-induced ApoE- / - mouse atherosclerosis model. The atherosclerosis model was established 17 weeks after modeling, and the control group was fed with ordinary feed for 17 weeks. After 17 weeks of modeling, the composition sample was continuously administered for 4 weeks to observe the formation of atherosclerotic plaques in the aorta of mice, and the plaque area was dissected and the percentage of plaques was statistically analyzed.

[0316] (2) Test results

[0317] The test results are shown in Table 5. The composition was extracted in the manner of Example 7. In the atherosclerosis treatment model, the red apple and cherry plum composition in the range of 40g / kg to 10g / kg was used in combination with high and low doses of positive drugs rosuvastatin and atorvastatin. Compared with the model group, the plaque in the arterial wall can be significantly reduced, and it has the effect of treating atherosclerosis. The composition has a synergistic effect when used in combination with the drug and when used alone (the calculation method refers to Experimental Example 1).

[0318] Table 5 The therapeutic effect of red apple and cherry plum combination combined with statins on atherosclerosis in mice

[0319]

[0320]

[0321] The significance in the above table is compared with the model group; *p<0.05, **p<0.01, ***p<0.001

[0322] Experimental Example 6: Evaluation of the therapeutic effect of the combination of the composition and other drugs on atherosclerosis in mice

[0323] (1) Test method

[0324] Wild-type C57 mice were used as the normal control group, and aspirin, clopidogrel, colchicine and probucol were used as positive drugs to investigate the anti-atherosclerotic efficacy (i.e., therapeutic effect) of the mixed composition sample in the high-fat diet-induced ApoE- / - mouse atherosclerosis model. The atherosclerosis model was established 17 weeks after modeling, and the control group was fed with ordinary feed for 17 weeks. After 17 weeks of modeling, the composition sample was continuously administered for 4 weeks to observe the formation of atherosclerotic plaques in the aorta of mice, and the plaque area was dissected and the percentage of plaques was statistically analyzed.

[0325] (2) Test results

[0326] The test results are shown in Table 6, which compares the anti-atherosclerotic effects of different positive drugs when used in combination with the composition. The composition was extracted in the manner of Example 7. In the atherosclerosis treatment model, the red apple and cherry plum composition in the range of 40g / kg to 10g / kg was used in combination with positive drugs aspirin, clopidogrel, colchicine and probucol. Compared with the model group, it can significantly reduce the plaque in the arterial wall and has the effect of treating atherosclerosis. The composition has a synergistic effect when used in combination with the drug and when used alone (the calculation method refers to Experimental Example 1).

[0327] Table 6 The therapeutic effect of red apple and cherry plum combination with other drugs on atherosclerosis in mice

[0328]

[0329]

[0330] The significance in the above table is compared with the model group; *p<0.05, **p<0.01, ***p<0.001

[0331] Experimental Example 7: Differences in the effects of different varieties of red apples and cherry plums on animal models

[0332] (1) Test method

[0333] Wild-type C57 mice were used as the normal control group, and rosuvastatin was used as the positive drug to investigate the anti-atherosclerotic efficacy (i.e., therapeutic effect) of the sample in the ApoE- / - mouse atherosclerosis model induced by a high-fat diet. The atherosclerosis model was established 17 weeks after modeling, and the control group was fed with ordinary feed for 17 weeks. After 17 weeks of modeling, the sample group was given continuous medication for 4 weeks to observe the formation of atherosclerotic plaques in the aorta of mice, and the plaque area was dissected and the percentage of plaques was calculated.

[0334] (2) Test results

[0335] As can be seen from Table 7, the inhibition rates of Hongxun No. 1 red flesh apple and Hongxun No. 3 red flesh apple are similar, with no significant difference. Therefore, it can be seen that there is no significant difference in the effects of different varieties of red flesh apples in animal models. Similarly, there is no significant difference in the effects of different varieties of cherry plums in animal models.

[0336] Table 7 The therapeutic effect of different varieties of red apple on atherosclerosis in mice

[0337] Group Plaque area% Compared with the model, the inhibition rate is % Model 39.66％ / Positive drug (rosuvastatin) 1.67mg / kg 25.87％** 34.77％ Red flesh apple (Hongxun No. 1) 20g / kg 32.89％* 17.07％ Red flesh apple (Hongxun No. 3) 20g / kg 32.56％* 17.90％

[0338] The significance in the above table is compared with the model group; *p<0.05, **p<0.01, ***p<0.001

[0339] Experimental Example 8: Differences in the effects of red apple and cherry plum compositions extracted by different methods in animal models

[0340] (1) Test method

[0341] The wild-type C57 mice were used as the normal control group, and rosuvastatin was used as the positive drug. The anti-atherosclerotic efficacy (i.e., therapeutic effect) of the mixed composition sample was investigated in the ApoE- / - mouse atherosclerosis model induced by a high-fat diet. The atherosclerosis model was established after 17 weeks of modeling, and the control group was fed with ordinary feed for 17 weeks. After 17 weeks of modeling, the composition sample was continuously administered for 4 weeks to observe the formation of atherosclerotic plaques in the aorta of mice, and the plaque area was dissected and the percentage of plaques was statistically analyzed.

[0342] (2) Test results

[0343] As can be seen from Table 8, the compositions prepared by the extraction methods with different ethanol concentrations in Example 7 and Example 8 all have good effects in treating atherosclerosis. It can be seen that the compositions of red apple and cherry plum with different extraction methods have good effects in animal models, and have better effects than using them alone.

[0344] Table 8 The therapeutic effect of red apple and cherry plum combination with different extraction methods on atherosclerosis in mice

[0345]

[0346]

[0347] The significance in the above table is compared with the model group; *p<0.05, **p<0.01, ***p<0.001

[0348] Experimental Example 9: Differences in the effects of red apple and cherry plum compositions extracted by different methods in animal models

[0349] (1) Test method

[0350] The wild-type C57 mice were used as the normal control group, and rosuvastatin was used as the positive drug. The anti-atherosclerotic efficacy (i.e., therapeutic effect) of the composition sample was investigated in the ApoE- / - mouse atherosclerosis model induced by a high-fat diet. The atherosclerosis model was established 17 weeks after modeling, and the control group was fed with ordinary feed for 17 weeks. After 17 weeks of modeling, the composition sample was continuously administered for 4 weeks to observe the formation of atherosclerotic plaques in the aorta of the mice, and the mice were dissected and the plaque area (percentage) was counted.

[0351] (2) Test results

[0352] As can be seen from Table 9, the compositions prepared by macroporous resin separation methods with different ethanol concentration gradient elution in Example 22 and Example 23 all have good effects in treating atherosclerosis. It can be seen that the red apple and cherry plum compositions with different extraction methods have good effects in animal models, and have better effects than using them alone.

[0353] Table 9 The therapeutic effect of red apple and cherry plum combination with different extraction methods on atherosclerosis in mice

[0354]

[0355]

[0356] The significance in the above table is compared with the model group; *p<0.05, **p<0.01, ***p<0.001

[0357] The above embodiments are only used to illustrate the technical solution of the present invention rather than to limit it. Any modification or equivalent replacement of the technical solution of the present invention without departing from the purpose and scope of the technical solution of the present invention should be included in the protection scope of the present invention.

Claims

1. Use of the composition in preparing a drug for preventing and / or treating atherosclerotic diseases, wherein the composition comprises the following first component and second component as active ingredients: The first component is a red apple extract ； and A second component, wherein the second component is a cherry plum extract; The weight ratio of the first component to the second component is 1:5-5:1; The first component and the second component are obtained by water extraction, and the water extraction comprises the following steps: Step 1: Remove the seeds or cores of red apples and cherry plums and then pulp them to directly obtain pulp; Step 2: Mix the pulp of red apple or cherry plum with water at a solid-liquid ratio of 1:2-10, add cellulase, extract at 10-100° C. for 30-100 min, and filter to obtain a pulp extract; Step 3: The red flesh apple or cherry plum pulp extract is concentrated by rotary evaporation at 30-60° C. to obtain a red flesh apple or cherry plum water extract; Step 4: Mix the extracted red apple or cherry plum water extracts in different weight ratios to prepare a composition, or first mix the red apple and cherry plum raw materials in the aforementioned different weight ratios in step 1 and then perform extraction.

2. The use according to claim 1, in, The weight ratio of the first component to the second component is 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 4:1 or 5:

1.

3. Use of the composition in preparing a drug for preventing and / or treating atherosclerotic diseases, wherein the composition comprises the following first component and second component as active ingredients: A first component, wherein the first component is a red apple extract; and A second component, wherein the second component is a cherry plum extract; The weight ratio of the first component to the second component is 1:2; The first component and the second component are obtained by alcohol leaching and extraction. The following steps are involved: Step 1: Remove the seeds or cores of red apples and cherry plums and then pulp them to directly obtain pulp; Step 2: mixing red apple or cherry plum pulp with 20%-60% ethanol at a solid-liquid ratio of 1:2-10, adding 0.2%-1% cellulase based on the weight of the pulp, extracting at 10-100° C. for 30-100 minutes, and filtering to obtain a pulp extract; Step 3: The red flesh apple or cherry plum pulp extract is concentrated by rotary evaporation at 30-60° C. to obtain a red flesh apple or cherry plum alcohol extract; the extract is evaporated to remove residual ethanol; Step 4: Mix the red apple or cherry plum alcohol extracts obtained by extraction in a weight ratio of 1:2 to prepare a composition, or first mix the red apple and cherry plum raw materials in a weight ratio of 1:2 in step 1 and then extract.

4. Use of the composition in preparing a drug for preventing and / or treating atherosclerotic diseases, wherein the composition comprises the following first component and second component as active ingredients: The first component is a red apple extract ； and A second component, wherein the second component is a cherry plum extract; The weight ratio of the first component to the second component is 1:2; The first component and the second component are obtained by macroporous resin separation, and the macroporous resin separation comprises the following steps: Step 1: preparing red apple and cherry plum extracts by water extraction as defined in claim 1; Step 2: Mix the extracts of red apple and cherry plum with 95%-100% ethanol, with the weight ratio of the extract to the ethanol being 1:2-10; the mixing time is 1-5 hours, and then filtering and separating to obtain alcohol precipitate juice and precipitate, and removing ethanol from the alcohol precipitate juice by rotary evaporation at a temperature of 30-60° C. to obtain alcohol precipitate clear juice; Step 3: using a macroporous adsorption resin to adsorb the alcohol precipitate clear juice, and gradient eluting with water and 20%-60% ethanol in sequence, collecting the eluent of each gradient respectively; the elution rate is 1-5 BV / h; Step 4: The above eluate is subjected to rotary evaporation to remove ethanol at a rotary evaporation temperature of 30-60° C. to obtain a macroporous resin separation sample.

5. The use according to any one of claims 1 to 4, in, The composition is in the form of freeze-dried powder, oral solution, capsule or tablet.

6. The use according to any one of claims 1 to 4, wherein the medicine is a Chinese patent medicine or a natural medicine.

7. The use according to any one of claims 1 to 4, in, The red flesh apple is selected from: Hongxun No. 1, Hongxun No. 3, Hongxun No. 4, Hongxun No. 6, Hongxun No. 8, and Red Love; Alternatively, the cherry plum is selected from red cherry plum, black cherry plum, purple cherry plum, purple-black cherry plum and yellow cherry plum.

8. Combination products, which include: A first product, wherein the first product is a composition as defined in any one of claims 1 to 5 and 7; The second product is a drug for preventing and / or treating atherosclerotic diseases.

9. The combination product according to claim 8, in, The atherosclerotic disease is aortic atherosclerosis; Alternatively, the drug for preventing and / or treating atherosclerotic diseases is selected from statins, aspirin, clopidogrel, colchicine, and probucol.

10. The combination product of claim 9, wherein the statin is selected from the group consisting of rosuvastatin, atorvastatin, and simvastatin.

11. The combination product according to any one of claims 8 to 10, in, The weight ratio of the first product to the second product is 1000:0.001-1000:33.

4.

12. Use of the combination product according to any one of claims 8 to 11 in the preparation of a medicament for preventing and / or treating atherosclerotic diseases.

13. The use according to claim 12, wherein the medicine is a Chinese patent medicine.

14. The use according to claim 12 or 13, in, The atherosclerotic disease is aortic atherosclerosis.

Images