WO2019041141A1 - 解纤维素拟杆菌在制备预防和/或治疗脂质代谢相关疾病制剂中的应用 - Google Patents

解纤维素拟杆菌在制备预防和/或治疗脂质代谢相关疾病制剂中的应用 Download PDF

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Publication number
WO2019041141A1
WO2019041141A1 PCT/CN2017/099522 CN2017099522W WO2019041141A1 WO 2019041141 A1 WO2019041141 A1 WO 2019041141A1 CN 2017099522 W CN2017099522 W CN 2017099522W WO 2019041141 A1 WO2019041141 A1 WO 2019041141A1
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Prior art keywords
composition
bacteroides cellulosilyticus
group
bacteroides
mammal
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PCT/CN2017/099522
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English (en)
French (fr)
Inventor
揭著业
粱穗莎
夏慧华
邹远强
肖亮
贾慧珏
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BGI Shenzhen Co Ltd
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BGI Shenzhen Co Ltd
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Priority to EP17923679.9A priority Critical patent/EP3677273B1/en
Priority to US16/642,702 priority patent/US11273186B2/en
Priority to PCT/CN2017/099522 priority patent/WO2019041141A1/zh
Priority to CN201810997577.2A priority patent/CN109419816B/zh
Publication of WO2019041141A1 publication Critical patent/WO2019041141A1/zh
Anticipated expiration legal-status Critical
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; PREPARATION THEREOF
    • A23C9/00—Milk preparations; Milk powder or milk powder preparations
    • A23C9/12—Fermented milk preparations; Treatment using microorganisms or enzymes
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; PREPARATION THEREOF
    • A23C9/00—Milk preparations; Milk powder or milk powder preparations
    • A23C9/152—Milk preparations; Milk powder or milk powder preparations containing additives
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K10/00—Animal feeding-stuffs
    • A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
    • A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
    • A23K10/18—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions of live microorganisms
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
    • A23K20/20—Inorganic substances, e.g. oligoelements
    • A23K20/24—Compounds of alkaline earth metals, e.g. magnesium
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K50/00—Feeding-stuffs specially adapted for particular animals
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K50/00—Feeding-stuffs specially adapted for particular animals
    • A23K50/50—Feeding-stuffs specially adapted for particular animals for rodents
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Preparation or treatment thereof
    • A23L2/52—Adding ingredients
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
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    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7016—Disaccharides, e.g. lactose, lactulose
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/702—Oligosaccharides, i.e. having three to five saccharide radicals attached to each other by glycosidic linkages
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/20—Milk; Whey; Colostrum
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A61K35/66—Microorganisms or materials therefrom
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/74—Bacteria
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    • A—HUMAN NECESSITIES
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    • A61K35/66—Microorganisms or materials therefrom
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    • A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis
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    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
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    • A61K36/06—Fungi, e.g. yeasts
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/04—Anorexiants; Antiobesity agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/06—Antihyperlipidemics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/11—Lactobacillus
    • A23V2400/113—Acidophilus
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/51—Bifidobacterium
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K2035/11—Medicinal preparations comprising living procariotic cells
    • A61K2035/115—Probiotics

Definitions

  • the present invention relates to the field of microbiology, and in particular to the use of Bacteroides cellulosilyticus for the prevention and/or treatment of diseases associated with lipid metabolism.
  • Atherosclerosis is the leading cause of coronary heart disease, cerebral infarction, and peripheral vascular disease.
  • Lipid metabolism disorder is the basis of atherosclerosis, which is characterized by the involvement of the affected arterial lesions from the intima, usually the accumulation of lipids and complex carbohydrates, hemorrhage and thrombosis, and then fibrous tissue hyperplasia and calcium deposition, and The gradual metamorphosis and calcification of the middle layer of the artery leads to thickening and hardening of the arterial wall and narrowing of the vascular lumen.
  • the lesion often involves the large and medium muscular arteries, and once developed enough to block the arterial lumen, the tissue or organ supplied by the artery will be ischemic or necrotic.
  • Atherosclerosis Since the appearance of lipids accumulated in the intima of the artery is yellow atheroma, it is called atherosclerosis. Atherosclerosis is caused by multiple factors, and the pathogenesis is complex. The main risk factors are hypertension, hyperlipidemia and massive smoking, as well as diabetes, obesity and genetic factors.
  • Atherosclerotic cardiovascular disease is the number one killer in developed countries and is becoming more common in developing countries.
  • China’s economy has developed rapidly, the people’s living standards have improved substantially, and the way of life is undergoing profound changes.
  • the rhythm of work and life is accelerating, the calorie of diet increases, but physical activity is reduced.
  • Cardiovascular risk factors such as obesity caused by unhealthy lifestyles have increased significantly, and the incidence of cardiovascular and cerebrovascular diseases has risen rapidly.
  • diseases such as stroke and myocardial infarction can lead to severe disability, reduced quality of life, and a heavy medical burden. High morbidity, mortality, and disability make cardiovascular and cerebrovascular diseases a major public health problem in China this century.
  • ACVD mainly includes coronary heart disease (CHD), stroke and peripheral arterial diseases.
  • CHD coronary heart disease
  • the causes of ACVD are multifaceted, and many related factors are related to lifestyle, including smoking, atherosclerotic diet, overweight or obesity, and sedentary, lack of physical activity.
  • the main cause of coronary heart disease is atherosclerosis, and the pathological changes of myocardial ischemia and necrosis. Its pathological pathogenesis is mainly due to lipid metabolism disorder, elevated blood lipids, lipid deposition and soaking in the inner wall of blood vessels, formation of vascular endothelial foam cells, and chronic inflammatory reaction of blood vessel wall, causing damage to coronary vascular endothelial function and morphology, forming blood vessels.
  • Endothelium plaque, further plaque rupture, thrombosis, stenosis or closure The plug causes coronary circulation disorder, myocardial ischemia, and myocardial cells are damaged, leading to cardiac insufficiency.
  • coronary circulation disorder myocardial ischemia, and myocardial cells are damaged, leading to cardiac insufficiency.
  • myocardial ischemia With the improvement of living standards, the incidence and mortality of coronary heart disease in China have also increased year by year, and there is a trend of rejuvenation.
  • Obesity is a group of common metabolic syndromes.
  • the external cause is mainly due to excessive diet and too little activity.
  • Calorie intake is more than calorie expenditure, making fat synthesis more important than obesity.
  • It is obese due to disorder of lipid metabolism.
  • the incidence of coronary heart disease and hypertension in obese patients is significantly higher than that in non-obese patients, and the incidence is generally 5 to 10 times that of non-obese patients, especially those with central waist obesity (male > 90 cm, female > 85 cm).
  • Obesity can cause cardiac hypertrophy, thickening of the posterior wall and interventricular septum, cardiac hypertrophy with increased blood volume, intracellular and intercellular fluid, ventricular end-diastolic pressure, pulmonary artery pressure and pulmonary capillary wedge pressure are increased, and some obese people have left ventricle Impaired function and obesity myocardial lesions.
  • the incidence of sudden death in obese patients is significantly increased, which may be related to hypertrophy of the heart muscle, arrhythmia caused by fat infiltration of the cardiac conduction system, and cardiac ischemia.
  • Modern medicine for the treatment of lipid metabolism related diseases especially atherosclerotic cardiovascular diseases (such as coronary heart disease) mainly includes drug treatment, interventional therapy, surgical treatment and so on.
  • oral Chinese medicine preparations for treating coronary heart disease are mainly based on Danshen preparations, such as Guanxin Danshen tablets (danshen, Sanqi, fragrant oil), Danshen tablets (danshen), Shuangdan oral liquid (danshen, peony bark), Dan Seven tablets (danshen, Sanqi), compound Danshen tablets (danshen, Sanqi, borneol), compound Danshen dripping pills (danshen, notoginseng, borneol), etc., all use activating blood as their main effect. Some of the above drugs contain borneol.
  • the borneol has the characteristics of aromatic walking, consuming gas and yang, long-term service and causing heart function reduction, and also stimulating the gastrointestinal tract.
  • Coronary heart disease has gastritis and stomach and twelve fingers. Patients with intestinal stains, esophagitis, or patients with debilitating constitution should not be used. At the same time, long-term use of compound Danshen tablets will lead to patients with bloating, fatigue and other potassium deficiency.
  • Interventional and surgical treatment have problems such as postoperative restenosis and high cost, and can not resolve the re-precipitation of intravascular lipids from the source to form new plaques, resulting in re-narrowing and occlusion of blood vessels, and poor blood supply to the atherosclerosis.
  • the problem of recurrence of sclerosing cardiovascular disease (such as coronary heart disease).
  • a first aspect of the invention provides the use of Bacteroides cellulosilyticus for the preparation of a composition or formulation for the prevention and/or treatment of a lipid metabolism related disorder.
  • the lipid metabolism-related disease is selected from the group consisting of atherosclerosis-related diseases, cardiovascular diseases, obesity, or a combination thereof.
  • Bacteroides cellulosilyticus is selected from the group consisting of Bacteroides cellulosilyticus DSM 14838, Bacteroides cellulosilyticus CL02T12C19, Bacteroides cellulosilyticus WH2, or a combination thereof.
  • the composition is selected from the group consisting of a food composition, a health care composition, a pharmaceutical composition, a beverage composition, a feed composition, or a combination thereof.
  • the composition is an oral preparation.
  • the composition is a liquid formulation, a solid formulation, or a semi-solid formulation.
  • the dosage form of the composition is selected from the group consisting of powders, powders, tablets, dragees, capsules, granules, suspensions, solutions, syrups, drops, and/or Or sublingual tablets.
  • the food composition comprises an emulsion preparation, a solution preparation, a powder preparation, or a suspension preparation.
  • the food composition comprises a dairy product, a milk powder, or an emulsion.
  • the liquid formulation is selected from the group consisting of a solution preparation or a suspension product.
  • the atherosclerosis-related disease is selected from the group consisting of coronary heart disease, coronary artery disease (CAD), atherosclerotic heart disease, atherosclerotic cardiovascular disease, ischemic Heart disease, or a combination thereof.
  • CAD coronary artery disease
  • atherosclerotic heart disease atherosclerotic cardiovascular disease
  • ischemic Heart disease ischemic Heart disease
  • the cardiovascular disease is selected from the group consisting of acute coronary syndrome, angina pectoris, arteriosclerosis, atherosclerosis, carotid atherosclerosis, cerebrovascular disease, cerebral infarction, congestive heart failure , congenital heart disease, coronary heart disease, coronary artery disease, coronary plaque stabilization, dyslipidemia, dyslipoproteinemia, endothelial dysfunction, familial hypercholesterolemia, familial complex high fat Blood, low alpha lipoproteinemia, hypertriglyceridemia, high beta lipoproteinemia, hypercholesterolemia, hypertension, hyperlipidemia, intermittent claudication, ischemia, ischemia reperfusion injury, Ischemic heart disease, myocardial ischemia, metabolic syndrome, multiple cerebral infarction dementia, myocardial infarction, obesity, peripheral vascular disease, reperfusion injury, restenosis, renal atherosclerosis Hardening, rheumatic heart disease, stroke, thrombotic disorder, transient ischemic attack, and with Alzheimer'sclerosis,
  • the formulation comprises a microecological formulation.
  • a second aspect of the invention provides the use of Bacteroides cellulosilyticus for the preparation of a composition or formulation for one or more uses selected from the group consisting of:
  • the improving vascular lesions in the mammal comprises ameliorating one or more indicators selected from the group consisting of blood viscosity, blood rheology, blood pressure, blood lipids (eg, triglycerides, total cholesterol, high density) Lipoprotein, low density lipoprotein), ST segment of electrocardiogram.
  • the reducing blood lipid levels in the mammal comprises lowering total cholesterol levels, triglyceride levels, low density lipoprotein levels, and/or blood viscosity levels in the blood.
  • the reduction in body weight of the mammal refers to a reduction in body weight of the experimental group of mammals by ⁇ 10%, preferably from 15% to 20%, compared to the model group of mammals.
  • the improving myocardial ischemia in a mammal comprises reducing a ST segment displacement of a myocardial ischemia in a mammal.
  • the mammal comprises a human or a non-human mammal.
  • the non-human mammal comprises a rodent such as a rat or a mouse.
  • a third aspect of the invention provides a composition for treating and/or preventing a cardiovascular disease, the composition comprising:
  • a fourth aspect of the invention provides a composition for preventing and/or treating obesity, the composition comprising:
  • a fifth aspect of the invention provides a method for treating and/or preventing an atherosclerosis-related disease a composition comprising:
  • a sixth aspect of the invention provides a composition for treating and/or preventing a lipid metabolism-related disease, the composition comprising:
  • Bacteroides cellulosilyticus is selected from the group consisting of Bacteroides cellulosilyticus DSM 14838, Bacteroides cellulosilyticus CL02T12C19, Bacteroides cellulosilyticus WH2, or a combination thereof.
  • the composition further comprises a growth factor (preferably, a milk growth factor).
  • a growth factor preferably, a milk growth factor
  • the composition is selected from the group consisting of a food composition, a health care composition, a pharmaceutical composition, a beverage composition, a feed composition, or a combination thereof.
  • the composition is an oral preparation.
  • the composition is a liquid formulation, a solid formulation, or a semi-solid formulation.
  • the dosage form of the composition is selected from the group consisting of powders, powders, tablets, dragees, capsules, granules, suspensions, solutions, syrups, drops, sublingual Tablets, or a combination thereof.
  • the food composition comprises an emulsion preparation, a solution preparation, a powder preparation, or a suspension preparation.
  • the food composition comprises a dairy product, a milk powder, or an emulsion.
  • the liquid formulation is selected from the group consisting of a solution preparation or a suspension product.
  • the composition contains 1 x 10-1 x 10 15 cfu/mL or cfu/g of Bacteroides cellulosilyticus, preferably 1 x 10 4 - 1 x 10 10 Cfu/mL or cfu/g of Bacteroides cellulosilyticus, based on the total volume or total weight of the composition.
  • the composition contains from 0.0001 to 99% by weight, preferably from 0.1 to 90% by weight, of the Bacteroides cellulosilyticus, based on the total weight of the composition.
  • the composition is a unit dosage form (one tablet, one capsule or one vial), and the mass of the composition in each unit dosage form is from 0.05 to 5 g, preferably from 0.1 to 1 g.
  • composition further contains other probiotics and/or prebiotics.
  • the probiotic is selected from the group consisting of lactic acid bacteria, bifidobacteria, Lactobacillus acidophilus, or a combination thereof.
  • the prebiotic is selected from the group consisting of oligofructose (FOS), galactooligosaccharide (GOS), xylooligosaccharide (XOS), oligofructose (LACT), and low soybean.
  • the composition further contains a substance (e.g., a protective agent) that helps maintain the viability of Bacteroides cellulosilyticus.
  • a substance e.g., a protective agent
  • the substance (such as a protective agent) which contributes to maintaining the viability of Bacteroides cellulosilyticus is selected from the group consisting of cysteine, glutathione, and butylhydroxyanisole.
  • the weight ratio (wt%) of the substance (such as a protective agent) which contributes to maintaining the activity of Bacteroides cellulosilyticus is 0.1-2%, preferably 0.5-1.5. %, more preferably, 0.5-1.0%, based on the total weight of the composition.
  • the amount of the substance (e.g., a protective agent) which contributes to maintaining the viability of Bacteroides cellulosilyticus is 1 mg to 20 mg, preferably 5 mg to 15 mg, based on 1 g of the composition. More preferably, 5 mg to 10 mg.
  • a seventh aspect of the present invention provides a process for producing a composition according to any of the third aspect of the present invention, wherein the method comprises the steps of:
  • the composition further contains a substance (e.g., a protective agent) that helps maintain the viability of Bacteroides cellulosilyticus.
  • a substance e.g., a protective agent
  • the substance (such as a protective agent) which contributes to maintaining the viability of Bacteroides cellulosilyticus is selected from the group consisting of cysteine, glutathione, and butylhydroxyanisole.
  • the composition is an oral preparation.
  • An eighth aspect of the invention provides a method of improving myocardial ischemia in a mammal comprising the steps of:
  • composition of any one of the third aspect of the invention to the sixth aspect of the invention is administered to the subject, thereby improving myocardial ischemia in the mammal.
  • the administration comprises oral administration.
  • the administration dose is 0.01 to 5 g / 50 kg body weight / day, preferably 0.1 to 2 g / 50 kg body weight / day.
  • the subject comprises a human or a non-human mammal.
  • non-human mammals include rodents (e.g., mice, rats), primates (e.g., monkeys).
  • the method is non-therapeutic and non-diagnostic.
  • a ninth aspect of the present invention provides a method of lowering blood fat and reducing body weight of a mammal, comprising the steps of:
  • composition of any one of the third aspect of the invention to the sixth aspect of the invention is administered to the subject, thereby lowering blood lipids and reducing body weight of the mammal.
  • the administration comprises oral administration.
  • the administration dose is 0.01 to 5 g / 50 kg body weight / day, preferably 0.1 to 2 g / 50 kg body weight / day.
  • the subject comprises a human or a non-human mammal.
  • non-human mammals include rodents (e.g., mice, rats), primates (e.g., monkeys).
  • the method is non-therapeutic and non-diagnostic.
  • a tenth aspect of the invention provides a method of improving vascular lesions in a mammal, comprising the steps of:
  • composition of any one of the third aspect of the invention to the sixth aspect of the invention is administered to the subject, thereby improving vascular lesions in the mammal.
  • the administration comprises oral administration.
  • the administration dose is 0.01 to 5 g / 50 kg body weight / day, preferably 0.1 to 2 g / 50 kg body weight / day.
  • the subject comprises a human or a non-human mammal.
  • the method is non-therapeutic and non-diagnostic.
  • the eleventh aspect of the present invention provides a method for treating and/or preventing a lipid metabolism-related disease Law, including steps:
  • composition of any one of the third aspect of the invention to the sixth aspect of the invention is administered to the subject, thereby treating and/or preventing a lipid metabolism-related disease.
  • the administration comprises oral administration.
  • the administration dose is 0.01 to 5 g / 50 kg body weight / day, preferably 0.1 to 2 g / 50 kg body weight / day.
  • the subject comprises a human or a non-human mammal.
  • the method is non-therapeutic and non-diagnostic.
  • Bacteroides cellulosilyticus has a disease for preventing and/or treating lipid metabolism (such as atherosclerosis-related diseases, cardiovascular diseases, The role of diseases such as obesity.
  • the active composition containing Bacteroides cellulosilyticus is fed to the test subject, and it is found that the composition can significantly reduce body weight, lower blood lipids, and improve myocardial ischemic diseases (such as reducing myocardial ischemia in mammals ST) Segment displacement), improve vascular lesions.
  • myocardial ischemic diseases such as reducing myocardial ischemia in mammals ST) Segment displacement
  • the term “contains” means that the various ingredients can be used together in the mixture or composition of the invention. Therefore, the terms “consisting essentially of” and “consisting of” are encompassed by the term “contains.”
  • growth factor includes milk growth factors, in particular, nutrients including vitamins, terpenoids, pyrimidines, or combinations thereof.
  • the vitamins include, but are not limited to: vitamin C, vitamin E, vitamin A, vitamin A precursor, vitamin B 6 , vitamin D 3 , vitamin K, folic acid, or a combination thereof;
  • the terpenoid includes, but is not limited to, a purine nucleoside, wherein the purine nucleoside comprises rhodium a 5'-phosphate of a purine nucleoside; the 5'-phosphate of the purine nucleoside is selected from the group consisting of inosinic acid (inosine-5'-phosphate; IMP) and guanylic acid (guanosine-5) '-phosphate ester; GMP), xanthosine (xanthine-5'-phosphate; XMP), adenylate (adenosine-5'-phosphate; AMP), or a combination thereof;
  • the pyrimidine species include all substances containing a pyrimidine structure.
  • reducing mammalian body weight and “controlling mammalian body weight gain” are used interchangeably and refer to the treatment of obesity-induced weight gain symptoms.
  • the term "improving vascular disease in a mammal” refers to treating a vascular lesion caused by atherosclerosis.
  • the improving vascular lesions in a mammal comprises ameliorating one or more indicators selected from the group consisting of blood viscosity, blood rheology, blood lipids (mainly triglycerides, total cholesterol, high density lipoprotein, Low density lipoprotein), ST segment of electrocardiogram.
  • ST segment displacement of myocardial ischemia means that myocardial ischemia can be diagnosed by electrocardiography, mainly showing ST segment depression or elevation.
  • the "Bacteroides cellulosilyticus of the present invention” is a strict anaerobic bacterium, and the cells are non-moving rods, having a length of 1.7 ⁇ m and a width of 0.9 ⁇ m, and Gram staining is negative and The cell wall is ultrastructured, with no heat-resistant spore formation, and the model bacteria are isolated from human feces. (See Robert C, Chassard C, Lawson P A, et al. Bacteroides cellulosilyticus sp. nov., a cellulolytic bacterium from the human gut microbial community [J]. International journal of systematic and evolutionary microbiology, 2007, 57(7): 1516-1520.)
  • Bacteroides cellulosilyticus is selected from the group consisting of Bacteroides cellulosilyticus DSM 14838, Bacteroides cellulosilyticus CL02T12C19, Bacteroides cellulosilyticus WH2, or a combination thereof.
  • the present invention provides an understanding of Bacteroides cellulosilyticus in the prevention and/or treatment of lipid metabolism related diseases (such as atherosclerosis-related diseases, cardiovascular diseases, obesity) Applications such as diseases.
  • the Bacteroides cellulosilyticus of the present invention has (i) lowering blood lipid levels in a mammal; (ii) reducing body weight in a mammal; (iii) improving myocardial ischemia in a mammal; and/or (iv) improving Vascular lesions in mammals.
  • SD rats are used as test rats, and a high-fat diet is used for hyperlipidemia modeling to obtain a hyperlipidemic model rat.
  • the hyperlipidemic model rats treated with Bacteroides cellulosilyticus showed significantly lower body weight than the untreated control group (model group), and various atherosclerosis-related diseases and cardiovascular diseases.
  • Disease-related indicators such as disease, obesity, such as blood lipid levels, total cholesterol levels, triglyceride levels, low-density lipoprotein levels, and blood viscosity levels are also significantly reduced.
  • SD rats are used as test rats, and SD rats are treated with pituitrin, and pituitary-treated SD rats treated with Bacteroides cellulosilyticus, Compared with the untreated control group (model group), the index related to cardiovascular disease (ST segment displacement of myocardial ischemia) was significantly reduced.
  • Bacteroides cellulosilyticus of the present invention can be used for the prevention and/or treatment of diseases related to lipid metabolism (such as atherosclerosis-related diseases, cardiovascular diseases, obesity and the like).
  • the present invention also provides a composition, preferably the composition comprises a food composition, a health care composition, a pharmaceutical composition, a beverage composition, or a feed composition, preferably a pharmaceutical composition.
  • the composition comprises (i) an effective amount of Bacteroides cellulosilyticus; and (ii) a pharmaceutically acceptable carrier.
  • the composition further comprises a growth factor (such as milk growth factor).
  • a growth factor such as milk growth factor.
  • the composition further comprises a probiotic selected from the group consisting of lactic acid bacteria, bifidobacteria, Lactobacillus acidophilus, or a combination thereof; and/or a prebiotic selected from the group consisting of oligofructose ( FOS), galactooligosaccharide (GOS), xylooligosaccharide (XOS), oligofructose (LACT), soy oligosaccharide (SOS), inulin, oligosaccharides, or combinations thereof.
  • FOS oligofructose
  • GOS galactooligosaccharide
  • XOS xylooligosaccharide
  • LACT oligofructose
  • SOS soy oligosaccharide
  • inulin oligosaccharides, or combinations thereof.
  • the composition further comprises a substance (e.g., a protective agent) selected from the group consisting of: cysteine, glutathione, butyl, which is useful for maintaining the viability of Bacteroides cellulosilyticus: Hydroxyanisole, dibutylmethyltoluene, tocopherol, bamboo leaf antioxidant, D-isoascorbic acid and its sodium salt, sodium ascorbate, calcium ascorbate, phospholipid, vitamin C (ascorbic acid), vitamin E, or a combination thereof.
  • a protective agent selected from the group consisting of: cysteine, glutathione, butyl, which is useful for maintaining the viability of Bacteroides cellulosilyticus: Hydroxyanisole, dibutylmethyltoluene, tocopherol, bamboo leaf antioxidant, D-isoascorbic acid and its sodium salt, sodium ascorbate, calcium ascorbate, phospholipid, vitamin C (ascorbic acid), vitamin E, or a combination thereof.
  • the weight ratio of the substance (e.g., protective agent) which contributes to maintaining the vitality of Bacteroides cellulosilyticus is 0.1-2%, preferably 0.5-1.5%, based on the total weight of the composition. Good place, 0.5-1.0%.
  • the composition is a liquid formulation, a solid formulation, or a semi-solid formulation.
  • the liquid formulation is selected from the group consisting of a solution product or a suspension product.
  • the dosage form of the composition is selected from the group consisting of powders, powders, tablets, dragees, capsules, granules, suspensions, solutions, syrups, drops, and/or Sublingual tablets.
  • composition of the present invention may be administered in the form of an oral liquid, a tablet, an injection, an orally disintegrating tablet, a lyophilized powder preparation or a capsule, preferably an enteric solvent type (e.g., a capsule), in the present invention, if there is no special
  • excipients, pharmaceutically acceptable vehicles and carriers used in the present invention are mainly selected according to the characteristics of the suitable bacteria or their metabolites and the specific administration mode required, and are advantageous for the bacteria or its metabolites to be smoothly passed through the stomach.
  • the drug is absorbed. These substances can be selected depending on the route of administration.
  • compositions of the present invention may further comprise any additional excipients in the excipients typically used in pharmaceutical formulations, for example, to stabilize the composition itself, or to readily disperse or impart a suitable taste.
  • inulin fructose, starch, xylooligosaccharide, silica, buffering agents, and flavoring agents are suitable examples.
  • the pharmaceutical preparation of the present invention may further comprise an auxiliary active ingredient.
  • the pharmaceutical composition of the present invention may further include a lubricant, a wetting agent, an emulsifier, a suspension stabilizer, a preservative, a sweetener, a perfume, and the like.
  • the pharmaceutical composition of the present invention can be produced in a casing formulation by various known methods, so that the active ingredient of the pharmaceutical composition, that is, the microorganism, can smoothly pass through the stomach without being destroyed by gastric acid.
  • the microorganism of the present invention can be used in the form of a capsule prepared by a conventional method.
  • a standard excipient is mixed with the cold-dried microorganism of the present invention to prepare a pellet, and then the pellet is filled into a gelatin capsule.
  • the microorganisms and medicaments of the present invention allow the use of excipients such as liquid gums, celluloses, silicates or mineral oils to prepare suspensions or dispersions, which suspensions or dispersions can be filled into soft gelatin capsules. in.
  • the pharmaceutical composition of the present invention can be made into an enteric tablet for oral use.
  • casing as used in this application includes coatings which are permitted for use with all conventional drugs which are not degraded by gastric acid but which are sufficiently decomposed in the small intestine and rapidly release the microorganism of the present invention.
  • the casing of the present invention is coated at about 16-30 mg per tablet, preferably 16-25 mg, more preferably 16-20 mg.
  • the thickness of the casing is 5 to 100 ⁇ m, and the thickness is preferably 20 to 80 ⁇ m.
  • the casing component is selected from conventional polymers known to the public.
  • Preferred casings of the present invention are cellulose acetate phthalate polymers or trimellitate polymers and copolymers of methacrylic acid (for example, containing 40% or more of methacrylic acid and containing methylcellulose phthalic acid) Preparation of a copolymer of hydroxypropyl acrylate or its ester derivative of methacrylic acid).
  • the cellulose acetate phthalate used in the casing of the present invention has a viscosity of about 45 to 90 cp, an acetyl content of 17 to 26%, and a phthalic acid content of 30 to 40%.
  • the cellulose acetate terephthalate used in the casing has a viscosity of about 5-21 cp and an acetaminophen content of 17-26%.
  • Cellulose acetate trimellitate is produced by Eastman Kodak Company and can be used in the casing material of the present invention.
  • the hydroxypropyl methylcellulose phthalate used in the casing of the present invention has a molecular weight of generally 20,000 to 130,000 Daltons, an ideal molecular weight of 80,000 to 100,000 Daltons, and a hydroxypropyl content of 5 to 10%.
  • the methoxy group content is 18-24%, and the phthaloyl group content is 21-35%.
  • HP50 The hydroxypropylmethylcellulose phthalate used in the casing of the present invention is HP50, which is produced by Shin-Etsu Chemidnl Co. Ltd., Japan. HP50 contains 6-10% hydroxypropyl, 20-24% methoxy, 21-27% propyl and has a molecular weight of 84,000 Daltons.
  • Another casing material is HP55, HP55 contains 5-9% hydroxypropyl methylcellulose phthalate, 18-22% methoxy, 27-35% phthalic acid, and its molecular weight is 78,000 Daltons.
  • the casing of the present invention is prepared by spraying a casing solution onto the core using conventional methods. All solvents in the enteric coating process are alcohols (e.g., ethanol), ketones (e.g., acetone), halogenated hydrocarbon compounds (e.g., dichloromethane), or combinations thereof. Softeners such as di-n-butyl phthalate and triacetin are added to the casing solution in a ratio of about 1 part to about 0.05 part or about 0.3 parts of softener.
  • the spraying method is preferably carried out continuously, and the amount of the sprayed material can be controlled according to the conditions employed for the coating.
  • the spray pressure can be adjusted at will, and in general, the desired result can be obtained at an average pressure of 1-1.5 Pa.
  • pharmaceutically effective amount is meant an amount that is functional or active to a human and/or animal and that is acceptable to humans and/or animals.
  • it may be prepared to contain 1 ⁇ 10 -1 ⁇ 10 20 cfu / ml or cfu / g (particularly, may contain 1 ⁇ 10 4 - 1 ⁇ 10 15 cfu / ml or cfu / g; more special A formulation of 1 x 10 6 - 1 x 10 11 cfu/ml or cfu/g) of Bacteroides cellulosilyticus may be contained.
  • the effective dose of Bacteroides cellulosilyticus used may vary depending on the mode of administration and the severity of the condition to be treated.
  • a dosage form suitable for internal administration comprising about 1 x 10-1 x 10 20 cfu/ml or cfu/g in intimate admixture with a solid or liquid pharmaceutically acceptable carrier (particularly, may contain 1 x 10 4 -1 x 10 15 cfu/ml or cfu/g; more particularly, may contain 1 ⁇ 10 6 -1 ⁇ 10 11 cfu/ml or cfu/g) of Bacteroides cellulosilyticus, or its fermentation-producing activity ingredient.
  • This dosage regimen can be adjusted to provide an optimal therapeutic response. For example, several separate doses may be administered per day, or the dose may be proportionally reduced, as is critical to the condition of the treatment.
  • the Bacteroides cellulosilyticus can be administered by oral or the like.
  • Solid carriers include: starch, lactose, dicalcium phosphate, microcrystalline cellulose, sucrose and kaolin, while liquid carriers include: medium, polyethylene glycol, nonionic surfactants and edible oils (such as corn oil, peanut oil). And sesame oil) as long as it is suitable for the characteristics of Bacteroides cellulosilyticus and the specific mode of administration required.
  • Adjuvants commonly used in the preparation of pharmaceutical compositions may also be advantageously included, such as flavoring agents, coloring agents, preservatives, and antioxidants such as vitamin E, vitamin C, BHT, and BHA.
  • compositions are solid compositions, especially tablets and solid filled or liquid filled capsules. Oral administration is preferred.
  • composition of the invention is administered to the individual for one or more administrations per day.
  • Dosage unit of administration means a dose that is formally separable and suitable for use in humans or all other mammalian individuals.
  • Each unit contains a pharmaceutically acceptable carrier and a therapeutically effective amount of a microorganism of the invention.
  • the amount administered varies depending on the severity of the patient's lipid metabolism-related diseases such as diseases such as atherosclerosis-related diseases, cardiovascular diseases, and obesity, the supplementary active components included, and the microorganisms used. Further, if possible, it can be administered separately and can be administered continuously if necessary. Therefore, the amount administered does not limit the invention.
  • the "composition" in the present invention means not only a medicine but also a functional food and a health supplement.
  • the composition comprises: a beverage, a food, a pharmaceutical, an animal feed, and the like.
  • a food composition comprising an effective amount of Bacteroides cellulosilyticus and a balance of a food acceptable carrier, said food composition
  • the dosage form is selected from the group consisting of a solid, a dairy, a solution, a powder, or a suspension.
  • the food composition may further comprise a growth factor (such as milk growth factor).
  • the composition further comprises a probiotic selected from the group consisting of lactic acid bacteria, bifidobacteria, Lactobacillus acidophilus, or a combination thereof; and/or a prebiotic selected from the group consisting of oligofructose ( FOS), galactooligosaccharide (GOS), xylooligosaccharide (XOS), oligofructose (LACT), soy oligosaccharide (SOS), inulin, oligosaccharides, or combinations thereof.
  • a probiotic selected from the group consisting of lactic acid bacteria, bifidobacteria, Lactobacillus acidophilus, or a combination thereof
  • a prebiotic selected from the group consisting of oligofructose ( FOS), galactooligosaccharide (GOS), xylooligosaccharide (XOS), oligofructose (LACT), soy oligosacc
  • the composition further comprises a substance (e.g., a protective agent) selected from the group consisting of: cysteine, glutathione, butyl, which is useful for maintaining the viability of Bacteroides cellulosilyticus: Hydroxyanisole, dibutylmethyltoluene, tocopherol, bamboo leaf antioxidant, D-isoascorbic acid and its sodium salt, sodium ascorbate, calcium ascorbate, phospholipid, vitamin C (ascorbic acid), vitamin E, or a combination thereof.
  • a protective agent selected from the group consisting of: cysteine, glutathione, butyl, which is useful for maintaining the viability of Bacteroides cellulosilyticus: Hydroxyanisole, dibutylmethyltoluene, tocopherol, bamboo leaf antioxidant, D-isoascorbic acid and its sodium salt, sodium ascorbate, calcium ascorbate, phospholipid, vitamin C (ascorbic acid), vitamin E, or a combination thereof.
  • composition of the composition is as follows:
  • composition of the composition is as follows:
  • the microecological preparation is a biological preparation containing probiotics and metabolites or a dietary supplement which can increase probiotics, and can achieve the purpose of improving human health by regulating and maintaining the micro-ecological balance in the intestinal tract. It mainly includes probiotics, prebiotics and synbiotics.
  • the microecological preparation comprises (a) a safe and effective amount of Bacteroides cellulosilyticus; and (b) a food acceptable carrier or a pharmaceutically acceptable carrier.
  • the formulation further comprises a growth factor (such as milk growth factor, preferably including a vitamin, an anthraquinone, and/or a pyrimidine).
  • the formulation further comprises a probiotic selected from the group consisting of lactic acid bacteria, bifidobacteria, Lactobacillus acidophilus, or a combination thereof; and/or a prebiotic selected from the group consisting of oligofructose (FOS) ), galactooligosaccharide (GOS), xylooligosaccharide (XOS), oligofructose (LACT), soy oligosaccharide (SOS), inulin, oligosaccharides, or combinations thereof.
  • a probiotic selected from the group consisting of lactic acid bacteria, bifidobacteria, Lactobacillus acidophilus, or a combination thereof
  • prebiotic selected from the group consisting of oligofructose (FOS) ), galactooligosaccharide (GOS), xylooligosaccharide (XOS), oligofructose (LACT), soy oligo
  • the composition further comprises a substance (e.g., a protective agent) selected from the group consisting of: cysteine, glutathione, butyl, which is useful for maintaining the viability of Bacteroides cellulosilyticus: Hydroxyanisole, dibutylmethyltoluene, tocopherol, bamboo leaf antioxidant, D-isoascorbic acid and its sodium salt, sodium ascorbate, calcium ascorbate, phospholipid, vitamin C (ascorbic acid), vitamin E, or a combination thereof.
  • a protective agent selected from the group consisting of: cysteine, glutathione, butyl, which is useful for maintaining the viability of Bacteroides cellulosilyticus: Hydroxyanisole, dibutylmethyltoluene, tocopherol, bamboo leaf antioxidant, D-isoascorbic acid and its sodium salt, sodium ascorbate, calcium ascorbate, phospholipid, vitamin C (ascorbic acid), vitamin E, or a combination thereof.
  • B. cellulolyticus is cultured in minced meat culture solution (DSMZ Medium 78) at 37 ° C for 48-72 h.
  • the method comprises ingesting a pharmaceutical composition, a food composition, a beverage composition, or a combination thereof of the present invention.
  • the subject includes mammals such as humans.
  • the method comprises: ingesting a pharmaceutical composition, a food composition, or an animal feed of the invention, or a combination thereof.
  • the subject is an animal, preferably a mouse, a rabbit.
  • the method comprises ingesting a pharmaceutical composition, a food composition, a beverage composition, or a combination thereof of the present invention.
  • the subject includes mammals such as humans.
  • the method comprises: ingesting a pharmaceutical composition, a food composition, or an animal feed of the invention, or a combination thereof.
  • the subject is an animal, preferably a mouse, a rabbit.
  • the method comprises ingesting a pharmaceutical composition, a food composition, a beverage composition, or a combination thereof of the present invention.
  • the subject includes mammals such as humans.
  • the method comprises: ingesting a pharmaceutical composition, a food composition, or an animal feed of the invention, or a combination thereof.
  • the subject is an animal, preferably a mouse, a rabbit.
  • the method comprises ingesting a pharmaceutical composition, a food composition, a beverage composition, or a combination thereof of the present invention.
  • the subject includes mammals such as humans.
  • the method comprises: ingesting a pharmaceutical composition, a food composition, or an animal feed of the invention, or a combination thereof.
  • the subject is an animal, preferably a mouse, a rabbit.
  • the Bacteroides cellulosilyticus of the present invention can significantly treat and/or prevent lipid metabolism-related diseases (such as atherosclerosis-related diseases, cardiovascular diseases, Obesity and other diseases).
  • lipid metabolism-related diseases such as atherosclerosis-related diseases, cardiovascular diseases, Obesity and other diseases.
  • the Bacteroides cellulosilyticus of the present invention can significantly reduce body weight and blood lipids (e.g., lower total cholesterol, triglycerides, low density lipoprotein, blood viscosity levels).
  • body weight e.g., lower total cholesterol, triglycerides, low density lipoprotein, blood viscosity levels.
  • the Bacteroides cellulosilyticus of the present invention can significantly improve myocardial ischemia.
  • Bacteroides cellulosilyticus of the present invention can significantly improve vascular lesions.
  • the Bacteroides cellulosilyticus of the present invention has obvious curative effects, small toxic and side effects, and greatly reduced cost.
  • Tongmai Jiangzhi Tablet is produced by Jilin Baitangtang Pharmaceutical Co., Ltd., the national medicine Zhunzi Z22024984.
  • Compound danshen Tablets is produced by Tianjin Tianshili (Liaoning) Pharmaceutical Co., Ltd., and the national medicine Zhunzi Z21020381.
  • Example 1 Therapeutic effect of Bacteroides cellulosilyticus in a rat model of hyperlipidemia
  • Bacteroides cellulosilyticus DSM 14838 (bacteria 1) was purchased from DSMZ (Leibniz-Institut DSMZ-Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH); Bacteroides cellulosilyticus CL02T12C19 (bacteria 2, obtained from Harvard Medical School, Coyne M J, Zitomersky N L, McGuire A M, et al. Evidence of extensive DNA transfer between bacteroidales species within the human gut [J].
  • Bacteroides cellulosilyticus WH2 Bacteroides 3, obtained from the University of Washington Medical Effects of diet on resource utilization by a model human gut microbiota containing Bacteroides cellulosilyticus WH2,a symbiont with an extensive glycobiome[J].PLoS Biology, 2013, 11(8): e1001637.
  • the microbial agents were placed in minced meat culture solution (DSMZ Medium78), anaerobic cultured at 37 ° C for 48-72 h, and the experiment was started after 16 S rDNA sequencing. The strain information is shown in Table 1.
  • Bacterial agent 1 Bacteroides cellulosilyticus DSM 14838 Bacterial agent 2 Bacteroides cellulosilyticus CL02T12C19 Bacterial agent 3 Bacteroides cellulosilyticus WH2
  • SD rats were purchased from the Guangdong Medical Laboratory Animal Center. Normal feeding, age 6 weeks. A total of 70 rats, the rat room environment is SPF (Specific Pathogen Free) level. After 1 week of adaptive feeding, the rats were randomly divided into 6 groups, including model control group, bacterial agent group 1, bacterial agent group 2, microbial agent group 3, Tongmai Jiangzhi tablets group (positive drugs for treating hyperlipidemia). And the normal control group, 10 in each group. Among them, the microbial agent group 1, the microbial agent group 2, the microbial agent group 3, the Tongmai Jiangzhi tablet group and the model control group were grouped and fed with high-fat diet for high-lipidemia model construction, and high-fat diet (including 78.8% basic diet).
  • 1% cholesterol, 10% egg yolk powder, 10% lard and 0.2% bile salt purchased from Nantong Trophy Feed Technology Co., Ltd.) for hyperlipidemia modeling, normal control group for feeding ordinary feed (purchased from Guangdongzhou Medicine) Laboratory Animal Center).
  • the concentration of viable bacteria was 10 9 cfu/ml, and that of Tongmai Jiangzhi tablets group was 0.6 g/kg per day in the group of bacteria group 1 and 2 groups.
  • the fresh bacterial liquid is replaced every other day, and the bacterial liquid is stored in an anaerobic storage condition at 4 °C. At the beginning of the experiment, while the high-fat diet was fed, the bacterial agent was intervened.
  • the bacteria in the group 1, 2, and 3 were intragastrically administered with 2 ml of the daily inoculum.
  • the model group was intragastrically administered with PBS (phosphate buffer) 2 ml daily.
  • the tablets were administered with 0.6 g/kg per day, and the body weight, status, and food intake of the rats were recorded weekly before and after the intervention.
  • the rats were sacrificed, the fat content was recorded, the serum was collected, and the serum lipid content, including total cholesterol (TC), triglyceride (TG), high-density lipoprotein (HDLC) and low density, was measured by reference to the kit instructions.
  • Lipoprotein (LDLC) Blood was taken from the last carotid artery and hemorrhagic examination was performed.
  • the data in Table 2 shows that as the experiment progressed, the body weight of the rats gradually increased, and the weight gain of the rats fed with high fat was higher than that of the rats in the normal group. At the 5th week, the rats of the normal group were normal. the body weight was significantly lower than the model group ( ⁇ P ⁇ 0.05), 9 weeks, the two sets of weight difference is even more significant ( ⁇ P ⁇ 0.0001). The body weight of the bacteria in the intervention group (bacteria 2, microbial agent 3) was different from that in the model group at the 7th week (*P ⁇ 0.05).
  • the results in Table 3 show that the intervention of the bacteria 1, 2, 3 can effectively control the levels of TC, TG, LDLC and blood viscosity in the blood (relative to the model group * P ⁇ 0.05), and the effect is better than the pulse drop Fat tablets.
  • the main components in blood lipids are cholesterol and triglycerides, and the increase in plasma cholesterol and triglyceride levels is closely related to the occurrence of atherosclerosis.
  • the microbial agents 1, 2, 3 can lower blood lipids and significantly reduce blood viscosity, thereby effectively preventing blood from being in a highly viscous, hypercoagulable state, improving blood rheology, and improving vascular lesions. Therefore, the microbial agents 1, 2, and 3 can reduce the related indicators of atherosclerosis-related diseases and cardiovascular diseases.
  • Example 2 Effect of Bacteroides cellulosilyticus on experimental myocardial ischemia in rats
  • the three groups of probiotics of the first embodiment are used as the medicine, and the preparation and dosage of the microbial agent are the same as those in the first embodiment, and the compound Danshen tablets are used as the control medicine, the vasopressin (the posterior pituitary injection), the national standard Word H31022259, specification 2ml: 6 units, produced by Shanghai Shangyi First Biochemical Pharmaceutical Co., Ltd.
  • SD rats were purchased from the Guangdong Medical Laboratory Animal Center. Normal feeding, age 6 weeks. A total of 70 rats, the rat room environment is SPF (Specific Pathogen Free) level.
  • SPF Specific Pathogen Free
  • the specific experimental results are shown in Table 4.
  • the three groups of bacteria provided by the present invention can effectively reduce the ST segment displacement of myocardial ischemia in rats (P ⁇ 0.01, *P ⁇ 0.05 relative to the model group), indicating the bacteria provided by the present invention.
  • the agent has a good improvement and protection effect on myocardial ischemia, and the effect is better than the compound danshen tablet group.
  • the raw materials are shown in Table 5.
  • the bacterial components in Formulations 1-3 are single bacterial components, respectively containing Bacteroides cellulosilyticus DSM 14838, Bacteroides cellulosilyticus CL02T12C19, and Bacteroides cellulosilyticus WH2.
  • formula 1-3 refers to a formula containing three kinds of bacteria, named formula 1, formula 2, formula 3.
  • Example 4 Pharmaceutical Composition Containing Bacteroides Cellulolyticum
  • the raw material ratio is shown in Table 6.
  • the bacterial components in Formulations 1-3 are single bacterial components, respectively containing Bacteroides cellulosilyticus DSM 14838, Bacteroides cellulosilyticus CL02T12C19, and Bacteroides cellulosilyticus WH2.
  • lactose, yeast powder and peptone are mixed uniformly with purified water, preheated to 60-65 ° C, homogenized at 20 Mpa, sterilized at 90 ° C for 20-30 minutes, cooled to 36-38 ° C, mixed with vitamin C, and connected.
  • Bacillus thuringiensis live bacteria (1-50 ⁇ 10 6 cfu/mL), fermented to pH 6.0 at 36-38 ° C, centrifuged, freeze-dried to a moisture content of less than 3%, ie, the preparation of frozen components containing bacteria Dry matter.
  • Example 5 Preparation of a drug containing Bacteroides disinfectant for treating a lipid metabolism related disease such as coronary heart disease
  • Preparation of drug or pharmaceutical dosage form adding 5 volumes (ml) of growth factor and 1 volume (ml) of a protective agent (such as vitamin C) to 100 volumes (ml) of Bacillus thuringiensis fermentation broth, fully The mixture is stirred and mixed, and then a starch adjuvant (such as maltodextrin) is added to prepare a drug or pharmaceutical dosage form containing Bacteroides.
  • a protective agent such as vitamin C

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Abstract

解纤维素拟杆菌(Bacteroides cellulosilyticus)在制备预防和/或治疗脂质代谢性相关疾病组合物或制剂中的应用，所述疾病是动脉粥样硬化相关疾病、心血管疾病和肥胖等。

Description

[根据细则37.2由ISA制定的发明名称]　解纤维素拟杆菌在制备预防和/或治疗脂质代谢相关疾病制剂中的应用 技术领域
本发明涉及微生物学领域，具体地，涉及解纤维素拟杆菌(Bacteroides cellulosilyticus)在预防和/或治疗脂质代谢性相关疾病方面的应用。
背景技术
动脉粥样硬化(atherosclerosis，AS)是冠心病、脑梗死、外周血管病的主要原因。脂质代谢障碍为动脉粥样硬化的病变基础，其特点是受累动脉病变从内膜开始，一般先有脂质和复合糖类积聚、出血及血栓形成，进而纤维组织增生及钙质沉着，并有动脉中层的逐渐蜕变和钙化，导致动脉壁增厚变硬、血管腔狭窄。病变常累及大中肌性动脉，一旦发展到足以阻塞动脉腔，则该动脉所供应的组织或器官将缺血或坏死。由于在动脉内膜积聚的脂质外观呈黄色粥样，因此称为动脉粥样硬化。动脉粥样硬化是多因素共同作用引起的，发病机制复杂，主要危险因素有高血压、高血脂和大量吸烟，还有糖尿病、肥胖和遗传因素等。
动脉粥样硬化性心血管疾病(atherosclerotic cardiovascular disease,ACVD)是发达国家的头号杀手，在发展中国家也日渐普遍。近年来，我国经济迅速发展，人民生活水平大幅度提高，生活方式正在发生深刻变化。工作和生活的节奏加快，饮食热量增加但体力活动减少。不良生活方式所引起的肥胖等心血管病危险因素显著增加，心脑血管病的发病率因而迅速上升。此外，脑卒中、心肌梗死等疾病可导致严重残疾，生活质量降低，医疗负担沉重。高发病率、死亡率、致残率使心脑血管病成为本世纪我国重大的公共卫生问题。
ACVD主要包括冠心病(coronary heart disease,CHD)、中风和周围动脉疾病。ACVD的病因是多方面的，许多相关因素都与生活方式有关，包括吸烟、致动脉粥样硬化饮食、超重或肥胖及惯于久坐、缺乏体力活动等。冠心病的主要病因是动脉粥样硬化，以及由此引起的心肌缺血、坏死的病理改变。其病理发病机制主要是由于脂质代谢紊乱使血脂升高，血管内壁脂质沉积、浸溶，血管内皮泡沫细胞形成，血管壁慢性炎性反应使冠脉血管内皮功能和形态的损伤，形成血管内皮粥样斑块，进一步出现斑块破裂，血栓形成，血管狭窄或闭 塞使冠脉血液循环障碍，心肌缺血，心肌细胞受损，导致心功能不全。随着生活水平提高，我国冠心病发病率和死亡率也逐年升高，并有年轻化趋势。
肥胖症是一组常见的代谢症群,外因以饮食过多而活动过少为主。热量摄入多于热量消耗，使脂肪合成增加是肥胖的物质基础。内因为脂质代谢紊乱而致肥胖。肥胖症患者并发冠心病、高血压的几率明显高于非肥胖者，其发生率一般5～10倍于非肥胖者，尤其腰围粗(男性>90cm，女性>85cm)的中心型肥胖患者。肥胖可致心脏肥大，后壁和室间隔增厚，心脏肥厚同时伴血容量、细胞内和细胞间液增加，心室舒张末压、肺动脉压和肺毛细血管楔压均增高，部分肥胖者存在左室功能受损和肥胖性心肌病变。肥胖患者猝死发生率明显升高，可能与心肌的肥厚、心脏传导系统的脂肪浸润造成的心律失常及心脏缺血有关。
现代医学对脂质代谢性相关疾病，尤其是动脉粥样硬化性心血管疾病(如冠心病)的治疗主要包括药物治疗、介入治疗、手术治疗等。
但是，药物治疗多有不同程度的毒副作用，且中成药其疗效大多不显著，治疗周期较长，不能彻底的治愈。目前市场上治疗冠心病的口服中药制剂主要以丹参制剂为主，如冠心丹参片(丹参、三七、降香油)、丹参片(丹参)、双丹口服液(丹参、牡丹皮)、丹七片(丹参、三七)、复方丹参片(丹参、三七、冰片)、复方丹参滴丸(丹参、三七、冰片)等，均以活血为其主要功效。且上述药物中有些含有冰片，冰片具有芳香走窜的特性，耗气伤阳，久服易致心脏功能减逞，同时对胃肠道刺激也较大，冠心病兼有胃炎、胃十二指肠渍窃、食道炎的患者，或属于虚寒体质的患者均不宜选用。同时长期服用复方丹参片会导致患者可出现腹胀、乏力等缺钾表现。
介入和手术治疗存在术后再狭窄和费用昂贵等问题，也不能从源头上解决血管内脂质等再沉淀形成新的斑块，致使血管重新变窄、堵塞，心肌供血不畅致动脉粥样硬化性心血管疾病(如冠心病)复发的问题。
因此，本领域迫切需要开发一种新的，无毒副作用的，用于治疗和/或预防脂质代谢性相关疾病(如动脉粥样硬化相关疾病、心血管疾病、肥胖)的药物。
发明内容
本发明的目的在于提供一种新的，无毒副作用的，用于治疗和/或预防脂 质代谢性相关疾病的药物。
本发明第一方面提供了一种解纤维素拟杆菌(Bacteroides cellulosilyticus)的用途，用于制备组合物或制剂，所述组合物或制剂用于预防和/或治疗脂质代谢性相关疾病。
在另一优选例中，所述脂质代谢性相关疾病选自下组：动脉粥样硬化相关疾病、心血管疾病、肥胖、或其组合。
在另一优选例中，所述解纤维素拟杆菌(Bacteroides cellulosilyticus)选自下组：Bacteroides cellulosilyticus DSM 14838、Bacteroides cellulosilyticus CL02T12C19、Bacteroides cellulosilyticus WH2、或其组合。
在另一优选例中，所述组合物选自下组：食品组合物、保健组合物、药物组合物、饮料组合物、饲料组合物、或其组合。
在另一优选例中，所述的组合物为口服制剂。
在另一优选例中，所述的组合物为液态制剂、固态制剂、半固态制剂。
在另一优选例中，所述的组合物的剂型选自下组：粉末剂、散剂、片剂、糖衣剂、胶囊剂、颗粒剂、悬浮剂、溶液剂、糖浆剂、滴剂、和/或舌下含片。
在另一优选例中，所述的食品组合物包括乳液制品、溶液制品、粉末制品、或悬浮液制品。
在另一优选例中，所述的食品组合物包括乳品、乳粉、或乳液。
在另一优选例中，所述的液态制剂选自下组：溶液制品或悬浮液制品。
在另一优选例中，所述动脉粥样硬化相关疾病选自下组：冠心病、冠状动脉疾病(CAD)、动脉粥样硬化性心脏病、动脉粥样硬化性心血管疾病、缺血性心脏病、或其组合。
在另一优选例中，所述心血管疾病选自下组：急性冠状动脉综合征、心绞痛、动脉硬化、动脉粥样硬化、颈动脉粥样硬化、脑血管疾病、脑梗塞、充血性心力衰竭、先天性心脏病、冠心病、冠状动脉疾病、冠脉斑块稳定(coronary plaque stabilization)、血脂异常症、异常脂蛋白血症、内皮功能障碍、家族性高胆固醇血症、家族性复合高脂血症、低α脂蛋白血症、高甘油三酯血症、高β脂蛋白血症、高胆固醇血症、高血压、高血脂、间歇性跛行、局部缺血、局部缺血再灌注损伤、缺血性心脏病、心肌缺血、代谢综合征、多发性脑梗塞痴呆、心肌梗塞、肥胖症、外周血管疾病、再灌注损伤、再狭窄、肾动脉粥样 硬化、风湿性心脏病、中风、血栓形成性紊乱、暂时性局部缺血发作以及与阿尔茨海默病、肥胖症、糖尿病、综合X、阳痿、多发性硬化症、帕金森病、炎性疾病有关的脂蛋白异常、或其组合。
在另一优选例中，所述制剂包括微生态制剂。
本发明第二方面提供了一种解纤维素拟杆菌(Bacteroides cellulosilyticus)的用途，用于制备组合物或制剂，所述组合物或制剂用于选自下组的一种或多种用途：
(i)降低哺乳动物的血脂水平；
(ii)降低哺乳动物的体重；
(iii)改善哺乳动物的心肌缺血；
(ⅳ)改善哺乳动物的血管病变。
在另一优选例中，所述改善哺乳动物的血管病变包括改善选自下组的一种或多种指标：血液粘度、血液流变、血压、血脂(如甘油三酯、总胆固醇、高密度脂蛋白、低密度脂蛋白)、心电图ST段。
在另一优选例中，所述降低哺乳动物的血脂水平包括降低血液中的总胆固醇水平、甘油三酯水平、低密度脂蛋白水平、和/或血粘度水平。
在另一优选例中，所述降低哺乳动物的体重指与模型组哺乳动物相比，实验组哺乳动物体重的降低幅度≥10％，较佳地，为15％-20％。
在另一优选例中，所述改善哺乳动物的心肌缺血包括降低哺乳动物心肌缺血ST段位移。
在另一优选例中，所述哺乳动物包括人或非人哺乳动物。
在另一优选例中，所述非人哺乳动物包括啮齿动物，如大鼠、小鼠。
本发明第三方面提供了一种用于治疗和/或预防心血管疾病的组合物，所述组合物包括：
(a)安全有效量的解纤维素拟杆菌(Bacteroides cellulosilyticus)；和
(b)食品上或药学上可接受的载体。
本发明第四方面提供了一种用于预防和/或治疗肥胖的组合物，所述组合物包括：
(a)安全有效量的解纤维素拟杆菌(Bacteroides cellulosilyticus)；和
(b)食品上或药学上可接受的载体。
本发明第五方面提供了一种用于治疗和/或预防动脉粥样硬化相关疾病的 组合物，所述组合物包括：
(a)安全有效量的解纤维素拟杆菌(Bacteroides cellulosilyticus)；和
(b)食品上或药学上可接受的载体。
本发明第六方面提供了一种用于治疗和/或预防脂质代谢性相关疾病的组合物，所述组合物包括：
(a)安全有效量的解纤维素拟杆菌(Bacteroides cellulosilyticus)；和
(b)食品上或药学上可接受的载体
在另一优选例中，所述解纤维素拟杆菌(Bacteroides cellulosilyticus)选自下组：Bacteroides cellulosilyticus DSM 14838、Bacteroides cellulosilyticus CL02T12C19、Bacteroides cellulosilyticus WH2、或其组合。
在另一优选例中，所述组合物还包括生长因子(较佳地，牛奶生长因子)。
在另一优选例中，所述组合物选自下组：食品组合物、保健组合物、药物组合物、饮料组合物、饲料组合物、或其组合。
在另一优选例中，所述的组合物为口服制剂。
在另一优选例中，所述的组合物为液态制剂、固态制剂、半固态制剂。
在另一优选例中，所述的组合物的剂型选自下组：粉末剂、散剂、片剂、糖衣剂、胶囊剂、颗粒剂、悬浮剂、溶液剂、糖浆剂、滴剂、舌下含片、或其组合。
在另一优选例中，所述的食品组合物包括乳液制品、溶液制品、粉末制品、或悬浮液制品。
在另一优选例中，所述的食品组合物包括乳品、乳粉、或乳液。
在另一优选例中，所述的液态制剂选自下组：溶液制品或悬浮液制品。
在另一优选例中，所述组合物含有1×10-1×1015cfu/mL或cfu/g的解纤维素拟杆菌(Bacteroides cellulosilyticus)，较佳地1×104-1×1010cfu/mL或cfu/g的解纤维素拟杆菌(Bacteroides cellulosilyticus)，按所述组合物的总体积或总重量计。
在另一优选例中，所述的组合物中，含有0.0001-99wt％，较佳地0.1-90wt％所述的解纤维素拟杆菌(Bacteroides cellulosilyticus)，以所述组合物的总重量计。在另一优选例中，所述的组合物为单元剂型(一片、一粒胶囊或一小瓶)，每个单元剂型中所述组合物的质量为0.05-5g，较佳地为0.1-1g。
在另一优选例中，所述的组合物还含有其它益生菌和/或益生元。
在另一优选例中，所述的益生菌选自下组：乳酸菌、双歧杆菌、嗜酸乳杆菌、或其组合。
在另一优选例中，所述的益生元选自下组：低聚果糖(FOS)、低聚半乳糖(GOS)、低聚木糖(XOS)、低聚乳果糖(LACT)、大豆低聚糖(SOS)、菊粉(Inulin)、寡聚糖、或其组合。
在另一优选例中，所述组合物还含有有助于保持解纤维素拟杆菌(Bacteroides cellulosilyticus)活力的物质(如保护剂)。
在另一优选例中，所述有助于保持解纤维素拟杆菌(Bacteroides cellulosilyticus)活力的物质(如保护剂)选自下组：半胱氨酸、谷胱甘肽、丁基羟基茴香醚、二丁基甲基甲苯、生育酚、竹叶抗氧化物、D-异抗坏血酸及其钠盐、抗坏血酸钠、抗坏血酸钙、磷脂、维生素C(抗坏血酸)、维生素E、或其组合。
在另一优选例中，所述有助于保持解纤维素拟杆菌(Bacteroides cellulosilyticus)活力的物质(如保护剂)的重量比(wt％)为0.1-2％，较佳地，0.5-1.5％，更佳地，0.5-1.0％，以所述组合物的总重计。
在另一优选例中，以组合物1g计，所述有助于保持解纤维素拟杆菌(Bacteroides cellulosilyticus)活力的物质(如保护剂)的含量为1mg-20mg，较佳地，5mg-15mg，更佳地，5mg-10mg。
本发明第七方面提供了一种本发明第三方面至本发明第六方面任一方面所述组合物的制法，其特征在于，包括步骤：
将(i)解纤维素拟杆菌(Bacteroides cellulosilyticus)、与(ii)食品上或药学上可接受的载体混合，从而形成本发明第三方面至本发明第六方面任一方面所述的组合物。
在另一优选例中，所述组合物还含有有助于保持解纤维素拟杆菌(Bacteroides cellulosilyticus)活力的物质(如保护剂)。
在另一优选例中，所述有助于保持解纤维素拟杆菌(Bacteroides cellulosilyticus)活力的物质(如保护剂)选自下组：半胱氨酸、谷胱甘肽、丁基羟基茴香醚、二丁基甲基甲苯、生育酚、竹叶抗氧化物、D-异抗坏血酸及其钠盐、抗坏血酸钠、抗坏血酸钙、磷脂、维生素C(抗坏血酸)、维生素E、或其组合。
在另一优选例中，所述的组合物为口服制剂。
本发明第八方面提供了一种改善哺乳动物的心肌缺血的方法，包括步骤：
给所述对象施用本发明第三方面至本发明第六方面任一方面所述的组合物，从而改善哺乳动物的心肌缺血。
在另一优选例中，所述的施用包括口服。
在另一优选例中，所述的施用剂量为0.01-5g/50kg体重/天，较佳地，0.1-2g/50kg体重/天。
在另一优选例中，所述的对象包括人或非人哺乳动物。
在另一优选例中，所述非人哺乳动物包括啮齿动物(如小鼠、大鼠)、灵长类动物(如猴)。
在另一优选例中，所述方法为非治疗性和非诊断性的。
本发明第九方面提供了一种降低血脂、降低哺乳动物的体重的方法，包括步骤：
给所述对象施用本发明第三方面至本发明第六方面任一方面所述的组合物，从而降低血脂、降低哺乳动物的体重。
在另一优选例中，所述的施用包括口服。
在另一优选例中，所述的施用剂量为0.01-5g/50kg体重/天，较佳地，0.1-2g/50kg体重/天。
在另一优选例中，所述的对象包括人或非人哺乳动物。
在另一优选例中，所述非人哺乳动物包括啮齿动物(如小鼠、大鼠)、灵长类动物(如猴)。
在另一优选例中，所述方法为非治疗性和非诊断性的。
本发明第十方面提供了一种改善哺乳动物的血管病变的方法，包括步骤：
给所述对象施用本发明第三方面至本发明第六方面任一方面所述的组合物，从而改善哺乳动物的血管病变。
在另一优选例中，所述的施用包括口服。
在另一优选例中，所述的施用剂量为0.01-5g/50kg体重/天，较佳地，0.1-2g/50kg体重/天。
在另一优选例中，所述的对象包括人或非人哺乳动物。
在另一优选例中，所述方法为非治疗性和非诊断性的。
本发明第十一方面提供了一种治疗和/或预防脂质代谢性相关疾病的方 法，包括步骤：
给所述对象施用本发明第三方面至本发明第六方面任一方面所述的组合物，从而治疗和/或预防脂质代谢性相关疾病。
在另一优选例中，所述的施用包括口服。
在另一优选例中，所述的施用剂量为0.01-5g/50kg体重/天，较佳地，0.1-2g/50kg体重/天。
在另一优选例中，所述的对象包括人或非人哺乳动物。
在另一优选例中，所述方法为非治疗性和非诊断性的。
应理解，在本发明范围内中，本发明的上述各技术特征和在下文(如实施例)中具体描述的各技术特征之间都可以互相组合，从而构成新的或优选的技术方案。限于篇幅，在此不再一一累述。
具体实施方式
本发明人经过广泛而深入的研究和实验，意外地发现，解纤维素拟杆菌(Bacteroides cellulosilyticus)具有预防和/或治疗脂质代谢性相关疾病(如动脉粥样硬化相关疾病、心血管疾病、肥胖等疾病)的作用。实验表明，将含有解纤维素拟杆菌(Bacteroides cellulosilyticus)的活性组合物饲喂食实验对象，发现该组合物能够显著降低体重，降低血脂，改善心肌缺血疾病(如降低哺乳动物的心肌缺血ST段位移)，改善血管病变。在此基础上，本发明人完成了本发明。
如本文所用，术语“含有”表示各种成分可一起应用于本发明的混合物或组合物中。因此，术语“主要由...组成”和“由...组成”包含在术语“含有”中。
如本文所用，术语“生长因子”包括牛奶生长因子，具体地，包括维生素类物质、嘌呤类物质、嘧啶类物质、或其组合的营养物质。
其中，所述维生素类物质包括(但并不限于)：维生素C、维生素E、维生素A、维生素A前体、维生素B6、维生素D3、维生素K、叶酸、或其组合；
所述嘌呤类物质包括(但并不限于)：嘌呤核苷，其中所述嘌呤核苷包括嘌 呤核苷的5’-磷酸酯；所述嘌呤核苷的5’-磷酸酯选自下组：肌苷酸(肌苷-5’-磷酸酯；IMP)、鸟苷酸(鸟苷-5’-磷酸酯；GMP)、黄苷酸(黄嘌呤核苷-5’-磷酸酯；XMP)、腺苷酸(腺苷-5’-磷酸酯；AMP)、或其组合；
所述嘧啶类物质包括所有含嘧啶结构的物质。
如本文所用，术语“降低哺乳动物体重”与“控制哺乳动物体重升高”可以互换使用，是指治疗肥胖引起的体重增加症状。
如本文所用，术语“改善哺乳动物的血管病变”是指治疗动脉粥样硬化引起的血管病变症状。
在本发明中，所述改善哺乳动物的血管病变包括改善选自下组的一种或多种指标：血液粘度、血液流变、血脂(主要是甘油三酯、总胆固醇、高密度脂蛋白、低密度脂蛋白)、心电图ST段。
心肌缺血ST段位移
如本文所用，术语“心肌缺血ST段位移”是指心肌缺血可以通过心电图检查来诊断，主要表现ST段压低或抬高。
本发明的解纤维素拟杆菌(Bacteroides cellulosilyticus)及其应用
如本文所用，所述“本发明的解纤维素拟杆菌(Bacteroides cellulosilyticus)”为严格厌氧菌，细胞为非运动型棒状，长为1.7μm，宽为0.9μm，革兰氏染色呈阴性和细胞壁超微结构，没有耐热的芽孢形成，模式菌分离自人粪便。(参见Robert C,Chassard C,Lawson P A,et al.Bacteroides cellulosilyticus sp.nov.,a cellulolytic bacterium from the human gut microbial community[J].International journal of systematic and evolutionary microbiology,2007,57(7):1516-1520.)
在一优选实施方式中，所述解纤维素拟杆菌(Bacteroides cellulosilyticus)选自下组：Bacteroides cellulosilyticus DSM 14838、Bacteroides cellulosilyticus CL02T12C19、Bacteroides cellulosilyticus WH2、或其组合。
本发明提供了解纤维素拟杆菌(Bacteroides cellulosilyticus)在预防和/或治疗脂质代谢性相关疾病(如动脉粥样硬化相关疾病、心血管疾病、肥胖 等疾病)方面的应用。本发明的解纤维素拟杆菌(Bacteroides cellulosilyticus)具有(i)降低哺乳动物的血脂水平；(ii)降低哺乳动物的体重；(iii)改善哺乳动物的心肌缺血；和/或(ⅳ)改善哺乳动物的血管病变。
根据本发明的一个优选例，采用SD大鼠作为试验大鼠，采用高脂饲料进行高血脂造模，从而获得高血脂模型大鼠。经解纤维素拟杆菌(Bacteroides cellulosilyticus)来治疗的高血脂模型大鼠，与未接受治疗的对照组(模型组)相比，其体重显著降低，并且各种动脉粥样硬化相关疾病、心血管疾病、肥胖等疾病相关的指标，如血脂水平、总胆固醇水平、甘油三酯水平、低密度脂蛋白水平和血粘度水平也显著降低。
根据本发明的另一个优选例采用SD大鼠作为试验大鼠，用垂体后叶素处理SD大鼠，经解纤维素拟杆菌(Bacteroides cellulosilyticus)来治疗的垂体后叶素处理的SD大鼠，与未治疗的对照组(模型组)相比，其与心血管疾病相关的指标(心肌缺血ST段位移)得到显著降低。
因此，本发明的解纤维素拟杆菌(Bacteroides cellulosilyticus)能够用以预防和/或治疗脂质代谢性相关疾病(如动脉粥样硬化相关疾病、心血管疾病、肥胖等相关疾病)。
组合物及其应用
本发明还提供了一种组合物，较佳地，所述组合物包括食品组合物、保健组合物、药物组合物、饮料组合物、或饲料组合物，优选地，为药物组合物。所述组合物包括(i)有效量的解纤维素拟杆菌(Bacteroides cellulosilyticus)；和(ii)药学上可接受的载体。
在一个优选例中，所述组合物还包括生长因子(如牛奶生长因子)。在一个优选例中，所述组合物还包括选自下组的益生菌：乳酸菌、双歧杆菌、嗜酸乳杆菌、或其组合；和/或选自下组的益生元：低聚果糖(FOS)、低聚半乳糖(GOS)、低聚木糖(XOS)、低聚乳果糖(LACT)、大豆低聚糖(SOS)、菊粉(Inulin)、寡聚糖、或其组合。
在一个优选例中，所述组合物还包括选自下组的有助于保持解纤维素拟杆菌(Bacteroides cellulosilyticus)活力的物质(如保护剂)：半胱氨酸、谷胱甘肽、丁基羟基茴香醚、二丁基甲基甲苯、生育酚、竹叶抗氧化物、D-异抗坏血酸及其钠盐、抗坏血酸钠、抗坏血酸钙、磷脂、维生素C(抗坏血酸)、维生素E、或其组合。 以组合物的总重计，所述有助于保持解纤维素拟杆菌(Bacteroides cellulosilyticus)活力的物质(如保护剂)的重量比为0.1-2％，较佳地，0.5-1.5％，更佳地，0.5-1.0％。
在一优选例中，所述的组合物为液态制剂、固态制剂、半固态制剂。
在一优选例中，所述的液态制剂选自下组：溶液制品或悬浮液制品。
在一优选例中，所述的组合物的剂型选自下组：粉末剂、散剂、片剂、糖衣剂、胶囊剂、颗粒剂、悬浮剂、溶液剂、糖浆剂、滴剂、和/或舌下含片。
本发明的组合物可以以口服液、片剂、针剂、口崩片、冻干粉制剂或胶囊的任一种形式给药，优选肠溶剂型(如胶囊)，在本发明中，如无特别说明，本发明所用的赋形剂、药物允许的媒介和载体主要是根据适合菌或其代谢产物特性和所需的特定给药方式进行选择，有利于菌或其代谢产物顺利通过胃而被给药者吸收。这些物质可根据给药途径进行选择。
本发明的组合物可进一步包含那些通常用于药物制剂的赋形剂中任何附加的赋形剂，为的是例如稳定组合物自身，或使其容易地分散或赋予其适宜的味道。
在所述赋形剂当中，菊粉、果糖、淀粉、低聚木糖、二氧化硅、缓冲试剂以及风味剂是合适的实例。
本发明中药物制剂可进一步包含辅助的活性组份。
乳糖、麦芽糊精、葡萄糖、蔗糖、山梨糖醇、甘露糖、淀粉、阿拉伯胶、磷酸钙、藻酸盐、明胶、硅酸钙、细结晶纤维素、聚乙烯吡咯烷酮(PVP)、纤维素、水、糖浆、甲基纤维素、羟基苯甲酸甲酯、羟基苯甲酸丙酯、滑石、硬脂酸镁或矿物油等都可用作本发明中药物组合物的载体、赋形剂或稀释剂等。
此外，本发明的药物组合物可进一步包括润滑剂、润湿剂、乳化剂、悬浮液稳定剂、防腐剂、甜味剂和香料等。本发明的药物组合物可通过多种公知的方法以肠衣制剂生产，以便于药物组合物的活性成分即微生物能顺利通过胃而不被胃酸所破坏。
另外，本发明的微生物可以常规方法制备的胶囊形式使用。例如，标准赋形剂和本发明的冷干微生物混合制成小球药丸，然后将药丸装填入明胶胶囊中。此外，本发明的微生物和药物允许使用的赋形剂如液体胶、纤维素、硅酸盐或矿物油等混合制作悬浮液或分散液，这种悬浮液或分散液可装入软的明胶胶囊中。
本发明的药物组合物可制成肠衣片供口服使用。本申请中的术语-“肠衣”，包括所有常规药物允许使用的包衣，这些包衣不被胃酸降解，但在小肠中能充分分解并快速释放出本发明的微生物。本发明的肠衣能在合成胃酸如pH＝1的HCl溶液中在36-38℃维持2小时以上，并优选在合成肠液如pH＝7.0的缓冲液中在1.0小时内分解。
本发明的肠衣为以每片约16-30mg进行包衣，较佳地16-25mg，更佳地16-20mg进行包衣。本发明中肠衣厚度为5-100μm，理想的厚度为20-80μm。肠衣成分选自己公开知晓的常规聚合物。
本发明优选的肠衣由纤维素乙酸邻苯二甲酸酯聚合物或偏苯三酸酯聚合物以及异丁烯酸的共聚物(例如，含有40％以上异丁烯酸和含有甲基纤维素邻苯二甲酸羟丙酯或其酯类衍生物的异丁烯酸的共聚物)制备。
本发明中肠衣所使用的纤维素乙酸邻苯二甲酸酯的粘度为约45-90cp，乙酰含量17-26％，邻苯二甲酸含量30-40％。用于肠衣中的纤维素乙酸偏苯二酸酯粘度为约5-21cp，乙酞含量17-26％。纤维素乙酸偏苯三酸酯由Eastman科达公司生产，可用于本发明中的肠衣材料。
用于本发明肠衣中的羟丙基甲基纤维素邻苯二甲酸酯，分子量一般为20,000-130,000道尔顿，理想分子量为80,000-100,000道尔顿，羟丙基含量为5-10％，甲氧基含量为18-24％，邻苯二甲酰基含量为21-35％。
用于本发明肠衣中的羟丙基甲基纤维素邻苯二甲酸酯为HP50，由日本Shin-Etsu Chemidnl Co.Ltd.生产。HP50含有6-10％羟丙基，20-24％甲氧基，21-27％的丙基，其分子量为84,000道尔顿。另一种肠衣物质为HP55，HP55含有5-9％的羟丙基甲基纤维素邻苯二甲酸酯，18-22％甲氧基，27-35％的邻苯二甲酸，其分子量为78,000道尔顿。
本发明肠衣如下制备：使用常规方法将肠衣溶液喷雾到核心上。该肠包衣方法中所有溶剂为醇类(如乙醇)、酮类(如丙酮)、卤代烃化合物(如二氯甲烷)、或其组合物。将软化剂如二-正丁基邻苯二甲酸酯和三乙酸甘油酯加入到肠衣溶液中，其比例为1份包衣物对约0.05份或约0.3份软化剂。喷雾方法优选连续执行，所喷雾的料量可根据包衣所采用的条件进行控制。喷雾压力可随意调节，一般而言，能在平均1-1.5帕压力下获得理想的结果。
说明书中“药物有效量”是指可对人和/或动物产生功能或活性的且可被人和/或动物所接受的量。比如，在本发明中，可制备含有1×10-1×1020cfu/ml 或cfu/g(特别的，可含有1×104-1×1015cfu/ml或cfu/g；更特别地，可含有1×106-1×1011cfu/ml或cfu/g)的解纤维素拟杆菌(Bacteroides cellulosilyticus)的制剂。
当用于制备药物组合物时，所用的解纤维素拟杆菌(Bacteroides cellulosilyticus)的有效剂量可随施用的模式和待治疗的疾病的严重程度而变化。适用于内服的剂量形式，包含与固态或液态药学上可接受的载体密切混合的约1×10-1×1020cfu/ml或cfu/g(特别的，可含有1×104-1×1015cfu/ml或cfu/g；更特别地，可含有1×106-1×1011cfu/ml或cfu/g)的解纤维素拟杆菌(Bacteroides cellulosilyticus)，或其发酵产生的活性成分。可调节此剂量方案以提供最佳治疗应答。例如，由治疗状况的迫切要求，可每天给予若干次分开的剂量，或将剂量按比例地减少。
所述的解纤维素拟杆菌(Bacteroides cellulosilyticus)可通过口服等途径给予。固态载体包括：淀粉、乳糖、磷酸二钙、微晶纤维素、蔗糖和白陶土，而液态载体包括：培养基、聚乙二醇、非离子型表面活性剂和食用油(如玉米油、花生油和芝麻油)，只要适合解纤维素拟杆菌(Bacteroides cellulosilyticus)特性和所需的特定给药方式。在制备药物组合物中通常使用的佐剂也可有利地被包括，例如调味剂、色素、防腐剂和抗氧化剂如维生素E、维生素C、BHT和BHA。
从易于制备和给药的立场看，优选的药物组合物是固态组合物，尤其是片剂和固体填充或液体填充的胶囊。口服给药是优选的。
将本发明组合物施用给所述个体，每天给药1次或多次。给药剂量单位表示其形式上能分开且适用于人类或其他所有哺乳动物个体的剂量。每一单位含有药物允许的载体和有效治疗量的本发明微生物。给药量随病人的脂质代谢性相关疾病(如动脉粥样硬化相关疾病、心血管疾病和肥胖等疾病)的严重程度、所包括的补充活性组份和所使用的微生物而变化。此外如可能，可分开给药，并且如需要可连续给药。因此，所述给药量不会对本发明造成限制。此外，本发明中的“组合物”不仅意味着药品而且表示可作为功能性食品和健康补充食品。在一个优选例中，所述组合物包括：饮料、食品、药品、动物饲料等。
在本发明的一个优选例中，还提供了一种食品组合物，它含有有效量的解纤维素拟杆菌(Bacteroides cellulosilyticus)以及余量的食品上可接受的载体，所述的食物组合物的剂型选自固体、乳品、溶液制品、粉末制品、或悬浮液制品。在一优选例中，所述食品组合物还可含有生长因子(如牛奶生长因子)。 在一优选例中，所述组合物还包括选自下组的益生菌：乳酸菌、双歧杆菌、嗜酸乳杆菌、或其组合；和/或选自下组的益生元：低聚果糖(FOS)、低聚半乳糖(GOS)、低聚木糖(XOS)、低聚乳果糖(LACT)、大豆低聚糖(SOS)、菊粉(Inul in)、寡聚糖、或其组合。在一个优选例中，所述组合物还包括选自下组的有助于保持解纤维素拟杆菌(Bacteroides cellulosilyticus)活力的物质(如保护剂)：半胱氨酸、谷胱甘肽、丁基羟基茴香醚、二丁基甲基甲苯、生育酚、竹叶抗氧化物、D-异抗坏血酸及其钠盐、抗坏血酸钠、抗坏血酸钙、磷脂、维生素C(抗坏血酸)、维生素E、或其组合。
在一优选例中，所述组合物的配方如下：
1×10-1×1020cfu/mL的解纤维素拟杆菌(Bacteroides cellulosilyticus)；以及食品上或药学上可接受的载体，和/或赋形剂。
在另一优选例中，所述组合物的配方如下：
1×106-1×1011cfu/mL的解纤维素拟杆菌(Bacteroides cellulosilyticus)；以及食品上或药学上可接受的载体，和/或赋形剂。
微生态制剂
微生态制剂是一种包含益生菌及代谢产物的生物制剂或者是可以增加益生菌的膳食补充剂，可以通过调节、维持肠道内微生态平衡，达到提高人体健康水平的目的。主要包括益生菌、益生元和合生元。
在本发明中，所述微生态制剂包括(a)安全有效量的解纤维素拟杆菌(Bacteroides cellulosilyticus)；以及(b)食品上可接受的载体或药学上可接受的载体。在一优选例中，所述制剂还包括生长因子(如牛奶生长因子，较佳地，包括维生素类物质、嘌呤类物质、和/或嘧啶类物质)。在一个优选例中，所述制剂还包括选自下组的益生菌：乳酸菌、双歧杆菌、嗜酸乳杆菌、或其组合；和/或选自下组的益生元：低聚果糖(FOS)、低聚半乳糖(GOS)、低聚木糖(XOS)、低聚乳果糖(LACT)、大豆低聚糖(SOS)、菊粉(Inulin)、寡聚糖、或其组合。在一个优选例中，所述组合物还包括选自下组的有助于保持解纤维素拟杆菌(Bacteroides cellulosilyticus)活力的物质(如保护剂)：半胱氨酸、谷胱甘肽、丁基羟基茴香醚、二丁基甲基甲苯、生育酚、竹叶抗氧化物、D-异抗坏血酸及其钠盐、抗坏血酸钠、抗坏血酸钙、磷脂、维生素C(抗坏血酸)、维生素E、或其组合。
解纤维素拟杆菌的培养方法
在本发明中，将解纤维素拟杆菌于肉末培养液(DSMZ Medium 78)中，37℃厌氧培养48-72h。
改善哺乳动物的心肌缺血的方法
在另一优选例中，所述方法包括：摄取本发明的药物组合物、食品组合物、饮料组合物、或其组合。所述实验对象包括哺乳动物，如人。
在另一优选例中，所述方法包括：摄取本发明的药物组合物、食品组合物、或动物饲料，或其组合。所述实验对象为动物，较佳地为鼠类，兔类。
降低血脂、降低哺乳动物的体重的方法
在另一优选例中，所述方法包括：摄取本发明的药物组合物、食品组合物、饮料组合物、或其组合。所述实验对象包括哺乳动物，如人。
在另一优选例中，所述方法包括：摄取本发明的药物组合物、食品组合物、或动物饲料，或其组合。所述实验对象为动物，较佳地为鼠类，兔类。
改善哺乳动物的血管病变的方法
在另一优选例中，所述方法包括：摄取本发明的药物组合物、食品组合物、饮料组合物、或其组合。所述实验对象包括哺乳动物，如人。
在另一优选例中，所述方法包括：摄取本发明的药物组合物、食品组合物、或动物饲料，或其组合。所述实验对象为动物，较佳地为鼠类，兔类。
治疗和/或预防脂质代谢性相关疾病的方法
在另一优选例中，所述方法包括：摄取本发明的药物组合物、食品组合物、饮料组合物、或其组合。所述实验对象包括哺乳动物，如人。
在另一优选例中，所述方法包括：摄取本发明的药物组合物、食品组合物、或动物饲料，或其组合。所述实验对象为动物，较佳地为鼠类，兔类。
本发明的主要优点包括：
(a)本发明的解纤维素拟杆菌(Bacteroides cellulosilyticus)能够显著治疗和/或预防脂质代谢性相关疾病(如动脉粥样硬化相关疾病、心血管疾病、 肥胖等疾病)。
(b)本发明的解纤维素拟杆菌(Bacteroides cellulosilyticus)能够显著降低体重、血脂(如降低总胆固醇、甘油三酯、低密度脂蛋白、血粘度水平)。
(c)本发明的解纤维素拟杆菌(Bacteroides cellulosilyticus)能够显著改善心肌缺血。
(d)本发明的解纤维素拟杆菌(Bacteroides cellulosilyticus)能够显著改善血管病变。
(e)本发明的解纤维素拟杆菌(Bacteroides cellulosilyticus)的疗效明显，毒副作用小，费用大大降低。
下面结合具体实施例，进一步阐述本发明。应理解，这些实施例仅用于说明本发明而不用于限制本发明的范围。下列实施例中未注明具体条件的实验方法，通常按照常规条件如Sambrook等人，分子克隆：实验室手册(New York：Cold Spring Harbor Laboratory Press，1989)中所述的条件，或按照《微生物：实验手册》(James Cappuccino和Natalie Sherman编，Pearson Education出版社)中所述的条件，或按照制造厂商所建议的条件。
如无特别说明，实施例所用的材料和试剂均为市售产品。
通脉降脂片由吉林百姓堂药业有限公司生产,国药准字Z22024984。
复方丹参片由天津天士力(辽宁)制药有限责任公司生产,国药准字Z21020381。
实施例1：解纤维素拟杆菌(Bacteroides cellulosilyticus)在高血脂症大鼠模型的治疗效果
Bacteroides cellulosilyticus DSM 14838(菌剂1)购自DSMZ(Leibniz-Institut DSMZ-Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH)；Bacteroides cellulosilyticus CL02T12C19(菌剂2,获自哈佛医学院，Coyne M J,Zitomersky N L,McGuire A M,et al.Evidence of extensive DNA transfer between bacteroidales species within the human gut[J].MBio,2014,5(3):e01305-14.)和Bacteroides cellulosilyticus WH2(菌剂3,获自华盛顿大学医学院，Effects of diet on resource utilization by a model human gut microbiota containing Bacteroides cellulosilyticus WH2,a symbiont with an extensive glycobiome[J].PLoS  biology,2013,11(8):e1001637.)。将菌剂分别放于肉末培养液(DSMZ Medium78)中，37℃厌氧培养48-72h，经16S rDNA测序鉴定无误后开始实验。菌株信息见表1。
表1菌株信息
编号 菌种名
菌剂1 Bacteroides cellulosilyticus DSM 14838
菌剂2 Bacteroides cellulosilyticus CL02T12C19
菌剂3 Bacteroides cellulosilyticus WH2
实验采用SD大鼠，购自广东省医学实验动物中心。正常饲养，鼠龄6周。总共70只大鼠，鼠房环境为SPF(Specific pathogen Free)级别。购买的大鼠适应性饲养1周后随机分为6组，包括模型对照组、菌剂1组、菌剂2组、菌剂3组、通脉降脂片组(治疗高血脂的阳性药物)和正常对照组，每组10只。其中菌剂1组、菌剂2组、菌剂3组、通脉降脂片组和模型对照组，分组后饲喂高脂饲料进行高血脂模型构建，采用高脂饲料(含78.8％基础饲料、1％胆固醇、10％蛋黄粉、10％猪油和0.2％胆盐，购自南通特洛菲饲料科技有限公司)进行高血脂造模，正常对照组饲喂普通饲料(购自广东省医学实验动物中心)。采用边造模边干预的方式，菌剂1组、菌剂2组和菌剂3组分别制备活菌浓度为109cfu/ml，通脉降脂片组每天给药0.6g/kg。为防止菌种失活甚至死亡，每隔一天更换新鲜菌液，菌液存放为4℃条件厌氧储存。实验开始，一边饲喂高脂饲料一边进行菌剂干预，菌剂1，2，3组每日灌胃菌液2ml，模型组每日灌胃PBS(磷酸盐缓冲液)2ml，通脉降脂片组每天给药0.6g/kg，分别在造模前后，干预前后每周记录大鼠的体重、状态、进食量等数据。实验结束后处死大鼠，记录脂肪含量，收集血清，参照试剂盒使用说明检测血清中血脂的含量，包括总胆固醇(TC)、甘油三酯(TG)、高密度脂蛋白(HDLC)和低密度脂蛋白(LDLC)。末次颈动脉取血，做血液流变学检查。
实验结果：
1、益生菌剂对高血脂模型大鼠体重的影响
表2、菌剂1,2,3分别对肥胖大鼠体重的影响
Figure PCTCN2017099522-appb-000001
Figure PCTCN2017099522-appb-000002
表2的数据显示，随着实验的进行，大鼠的体重逐渐升高，其中高脂饲喂的大鼠体重增加幅度高于正常组大鼠体重的增加，在第5周，正常组大鼠的体重显著低于模型组(▲P<0.05)，第9周，这两组的体重差异更加显著(▲▲▲P<0.0001)。菌剂(菌剂2，菌剂3)干预组大鼠的体重在第7周同模型组开始有了差异(*P<0.05)，在实验的第9周，三组菌剂干预组大鼠体重较模型组的差异也更加显著(**P<0.01),且效果均优于通脉降脂片，说明菌剂1,2,3的干预可以有效控制大鼠体重升高。
2、益生菌剂对肥胖模型大鼠血脂的影响
表3、菌剂1,2,3分别对肥胖大鼠血脂的影响
Figure PCTCN2017099522-appb-000003
Figure PCTCN2017099522-appb-000004
表3的结果显示，菌剂1,2,3的干预可以有效的控制血液中TC，TG，LDLC以及血粘度的水平(相对于模型组*P<0.05),且效果均优于通脉降脂片。其中血脂中的主要成份是胆固醇和甘油三酯，血浆中胆固醇和甘油三脂水平的升高与动脉粥样硬化的发生有密切的关系。此外，菌剂1,2,3可以降低血脂，并能够显著降低血液粘度，从而有效地防止血液处于高黏、高凝状态，改善血液流变学，改善血管病变。因此，菌剂1,2,3可以降低动脉粥样硬化相关疾病、心血管疾病的相关指标。
实施例2：解纤维素拟杆菌(Bacteroides cellulosilyticus)在对大鼠实验性心肌缺血的影响
1、实验材料
(1)受试药物
本实例采用实施例1的3组益生菌菌剂作为药物，菌剂的制备和用量同实施例1，同时采用复方丹参片作为对照药物，垂体后叶素(垂体后叶注射液)，国药准字H31022259，规格2ml:6单位，由上海上药第一生化药业有限公司生产。
(2)实验动物
采用SD大鼠，购自广东省医学实验动物中心。正常饲养，鼠龄6周。总共70只大鼠，鼠房环境为SPF(Specific pathogen Free)级别。
(3)实验过程
对脑垂体后叶素所致大鼠急性心肌缺血的影响
取SD大鼠50只，雌雄各半，体重180～220克，随机分为模型组、复方丹参片组，菌剂1组，菌剂2组，菌剂3组，每组10只大鼠。复方丹参片组和菌剂组每天灌胃给药1次，复方丹参片组每天给药2g/kg，菌剂组每天灌胃菌液2ml(菌浓度为109cfu/ml)，模型组给与等容量的PBS，连续10天。末次给药后lh，记录标I导联心电图，由舌下iv(静脉注射)脑垂体后叶素1单位/kg，容量为1m l/kg，于15S注射完毕，然后立即描记5S、15S、30S、lmin、2min、3min和5min的心电图变化，测量比较给垂体后叶素前后ST段抬 高的变化，以组间t检验进行统计分析。具体实验结果见表4所示。
表4.菌剂和复方丹参片对垂体后叶素致大鼠心肌缺血ST段位移的影响
分组 ST位移(mv)(±S)
模型组 0.061±0.012
复方丹参片组 0.032±0.016*
菌剂1组 0.028±0.009**
菌剂2组 0.027±0.011**
菌剂3组 0.021±0.012**
通过表4中统计显示，本发明提供的3组菌剂均能有效降低大鼠心肌缺血ST段位移(相对于模型组**P<0.01，*P<0.05)，表明本发明提供的菌剂对心肌缺血具有很好的改善和保护作用，且效果优于复方丹参片组。
实施例3含解纤维素拟杆菌的食品组合物
原料配比如表5。
表5
原料 质量百分比(％)
菌组分 0.5
牛奶 90.0
白糖 9.0
维生素C 0.5
配方1-3中的菌组分为单菌组分，分别含有Bacteroides cellulosilyticus DSM 14838、Bacteroides cellulosilyticus CL02T12C19、Bacteroides cellulosilyticus WH2。其中，配方1-3指的是分别含有三种菌的配方，命名为配方1，配方2，配方3。
按照上述配方比例混合牛奶、白糖，搅拌至完全混合，预热，20Mpa压力均质，90℃左右杀菌5-10分钟，冷却至40-43℃，混入保护剂维生素C，接种1-100×106cfu/g的菌组分，即制成含菌组分(如解纤维素拟杆菌)的食品组合物。
实施例4含解纤维素拟杆菌的药物组合物
原料配比见表6。
表6
原料 质量百分比(％)
菌组分 1.0％
乳糖 2.0％
酵母粉 2.0％
蛋白胨 1.0％
纯净水 93.5％
维生素C 0.5％
配方1-3中的菌组分为单菌组分，分别含有Bacteroides cellulosilyticus DSM 14838、Bacteroides cellulosilyticus CL02T12C19、Bacteroides cellulosilyticus WH2。
按照比例将乳糖、酵母粉、蛋白胨以纯净水混合均匀，预热到60-65℃，20Mpa压力均质，90℃左右杀菌20-30分钟，冷却至36-38℃，混入维生素C，接入解纤维素拟杆菌活菌(1-50×106cfu/mL)，36-38℃发酵至pH值为6.0，离心，冷冻干燥至水份含量小于3％，即制备含菌组分的冷冻干燥物。称取0.5克含菌组分的冷冻干燥物与麦芽糊精、保护剂(如维生素C)等量混合后装入胶囊中，即制成含菌组分(如解纤维素拟杆菌)的药物组合物。
实施例5一种用于治疗脂质代谢性相关疾病(如冠心病)的含解纤维素拟杆菌的药物的制备
1、菌液准备：将解纤维素拟杆菌(1×109cfu/ml)进行厌氧培养，厌氧培养基采用PYG培养基，经过37℃厌氧发酵2-3天。
2、生长因子制备：将脱脂牛奶、酪蛋白进行混合、离心、超滤获得牛奶生长因子粗提物(含有维生素类物质、嘌呤类物质、和/或嘧啶类物质的营养成分)。
3、药物或药物剂型制作：将5体积(ml)生长因子和1体积(ml)的保护剂(如维生素C)加入到100体积(ml)的解纤维素拟杆菌发酵的菌液中，充分搅拌混匀，然后加入淀粉辅料(如麦芽糊精)，从而制备得到含有解纤维素拟杆菌的药物或药物剂型。
在本发明提及的所有文献都在本申请中引用作为参考，就如同每一篇文献被单独引用作为参考那样。此外应理解，在阅读了本发明的上述讲授内容之后，本领域技术人员可以对本发明作各种改动或修改，这些等价形式同样落于本申请所附权利要求书所限定的范围。

Claims (18)

  1. 一种解纤维素拟杆菌(Bacteroides cellulosilyticus)的用途，其特征在于，用于制备组合物或制剂，所述组合物或制剂用于预防和/或治疗脂质代谢性相关疾病。
  2. 如权利要求1所述的用途，其特征在于，所述脂质代谢性相关疾病选自下组：动脉粥样硬化相关疾病、心血管疾病、肥胖、或其组合。
  3. 如权利要求1所述的用途，其特征在于，所述解纤维素拟杆菌(Bacteroides cellulosilyticus)选自下组：Bacteroides cellulosilyticus DSM 14838、Bacteroides cellulosilyticus CL02T12C19、Bacteroides cellulosilyticus WH2、或其组合。
  4. 一种解纤维素拟杆菌(Bacteroides cellulosilyticus)的用途，其特征在于，用于制备组合物或制剂，所述组合物或制剂用于选自下组的一种或多种用途：
    (i)降低哺乳动物的血脂水平；
    (ii)降低哺乳动物的体重；
    (iii)改善哺乳动物的心肌缺血；
    (ⅳ)改善哺乳动物的血管病变。
  5. 一种用于治疗和/或预防心血管疾病的组合物，其特征在于，所述组合物包括：
    (a)安全有效量的解纤维素拟杆菌(Bacteroides cellulosilyticus)；和
    (b)食品上或药学上可接受的载体。
  6. 一种用于预防和/或治疗肥胖的组合物，其特征在于，所述组合物包括：
    (a)安全有效量的解纤维素拟杆菌(Bacteroides cellulosilyticus)；和
    (b)食品上或药学上可接受的载体。
  7. 一种用于治疗和/或预防动脉粥样硬化相关疾病的组合物，其特征在于，所述组合物包括：
    (a)安全有效量的解纤维素拟杆菌(Bacteroides cellulosilyticus)；和
    (b)食品上或药学上可接受的载体。
  8. 一种用于治疗和/或预防脂质代谢性相关疾病的组合物，其特征在于，所述组合物包括：
    (a)安全有效量的解纤维素拟杆菌(Bacteroides cellulosilyticus)；和
    (b)食品上或药学上可接受的载体。
  9. 如权利要求5-8任一项所述的组合物，其特征在于，所述的组合物还含有其它益生菌和/或益生元。
  10. 如权利要求9所述的组合物，其特征在于，所述的益生菌选自下组：乳酸菌、双歧杆菌、嗜酸乳杆菌、或其组合。
  11. 如权利要求9所述的组合物，其特征在于，所述的益生元选自下组：低聚果糖(FOS)、低聚半乳糖(GOS)、低聚木糖(XOS)、低聚乳果糖(LACT)、大豆低聚糖(SOS)、菊粉(Inulin)、寡聚糖、或其组合。
  12. 如权利要求5-8任一项所述的组合物，其特征在于，所述组合物还含有有助于保持解纤维素拟杆菌(Bacteroides cellulosilyticus)活力的物质。
  13. 一种权利要求5-8任一项所述组合物的制法，其特征在于，包括步骤：
    将(i)解纤维素拟杆菌(Bacteroides cellulosilyticus)、与(ii)食品上或药学上可接受的载体混合，从而形成权利要求5-8任一项所述的组合物。
  14. 如权利要求13所述的制法，其特征在于，所述组合物还含有有助于保持解纤维素拟杆菌(Bacteroides cellulosilyticus)活力的物质。
  15. 一种改善哺乳动物的心肌缺血的方法，其特征在于，包括步骤：
    给所述对象施用权利要求5-8任一项所述的组合物，从而改善哺乳动物的心肌缺血。
  16. 一种降低血脂、降低哺乳动物的体重的方法，其特征在于，包括步骤：
    给所述对象施用权利要求5-8任一项所述的组合物，从而降低血脂、降低哺乳动物的体重。
  17. 一种改善哺乳动物的血管病变的方法，其特征在于，包括步骤：
    给所述对象施用权利要求5-8任一项所述的组合物，从而改善哺乳动物的血管病变。
  18. 一种治疗和/或预防脂质代谢性相关疾病的方法，其特征在于，包括步骤：
    给所述对象施用权利要求5-8任一项所述的组合物，从而治疗和/或预防脂质代谢性相关疾病。
PCT/CN2017/099522 2017-08-29 2017-08-29 解纤维素拟杆菌在制备预防和/或治疗脂质代谢相关疾病制剂中的应用 Ceased WO2019041141A1 (zh)

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EP17923679.9A EP3677273B1 (en) 2017-08-29 2017-08-29 Bacteroides cellulosityticus for use in preventing and/or treating atherosclerosis and obesity
US16/642,702 US11273186B2 (en) 2017-08-29 2017-08-29 Application of bacteroides cellulosityticus in preparing a preparation for preventing and/or treating lipid metabolism related diseases
PCT/CN2017/099522 WO2019041141A1 (zh) 2017-08-29 2017-08-29 解纤维素拟杆菌在制备预防和/或治疗脂质代谢相关疾病制剂中的应用
CN201810997577.2A CN109419816B (zh) 2017-08-29 2018-08-29 解纤维素拟杆菌在预防和/或治疗心脏疾病方面的应用

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CN116036134A (zh) * 2022-11-30 2023-05-02 广东省人民医院 一种用于预防和/或治疗心血管疾病的细菌组合物及应用

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