Pharmaceutical composition and application thereof

Abstract

The invention belongs to the technical field of medicines, and particularly relates to a pharmaceutical composition and application thereof. The pharmaceutical composition comprises (a) an OB01 strain or a filial generation strain or a subcloning strain or a culture supernatant or a preparation thereof, and (b) other drugs for treating metabolic diseases, the bacterial strain OB01 is the bifidobacterium bifidum ibiome001, and the bacterial strain OB01 is the bifidobacterium bifidum ibiome001. After the OB01 is combined with multiple positive drugs, blood sugar, blood fat and obesity are improved to different extents, a synergistic effect is achieved, the curative effect can be improved, the drug dosage can be reduced, intestinal flora balance can be adjusted, adverse reactions caused by common drugs can be reduced, and the drug resistance of the common drugs can be reduced by adjusting the metabolic process of an organism. The OB01 is combined with common positive drugs for use, a safer and more effective treatment scheme is provided for patients, and important clinical significance and application value are achieved.

Description

Technical Field

[0001] The present invention belongs to the field of medical technology, and in particular relates to a pharmaceutical composition and use thereof. Background Art

[0002] In the first 100 years after probiotics were discovered and used, they were only used as food or dietary supplements. In recent years, with the development of sequencing technology and the in-depth study of microbiome, people have gradually realized the great potential of microorganisms in treating diseases. Live biotherapeutic products (LBP), also known as "second-generation probiotics", have the advantages of high safety, good efficacy and low cost, and are expected to become a "dark horse" in the field of innovative drug research and development in the future.

[0003] CN115948273A discloses a new strain of Bifidobacterium bifidum ( Bifidobacterium bifidum )ibiome001 (OB01), this strain can significantly reduce the body weight and white fat weight of obese and diabetic mice induced by high-fat diet, and increase the expression of lipolytic genes ATGL and HSL; and significantly reduce the fasting blood glucose, area under the oral glucose tolerance curve, plasma glycosylated hemoglobin and insulin content of mice. It is a potential functional strain for the treatment of metabolic syndrome such as obesity and diabetes, and its effect is better than the combination with other Bifidobacterium bifidum.

[0004] At present, although common drugs for treating metabolic diseases can control the disease to a certain extent, they have many disadvantages: (1) Adverse reactions: For example, metformin, common adverse reactions include gastrointestinal reactions (nausea, diarrhea, aversion to oil, etc.), abnormal liver function, etc.; statins, common adverse reactions include muscle pain, abnormal liver function, etc.; diuretics, calcium channel blockers, etc., common adverse reactions include hypotension, increased heart rate, electrolyte imbalance, etc.; (2) Drug resistance: Long-term use of certain drugs may lead to drug resistance, which reduces the efficacy of the drug and makes the disease difficult to control; (3) Drug interactions: Patients with metabolic diseases often need to take multiple drugs at the same time. The interactions between drugs may lead to reduced efficacy or increased adverse reactions.

[0005] Given the many drawbacks of current common drugs for treating metabolic diseases, the combination of living biological drugs with these drugs has become a new treatment strategy. Summary of the invention

[0006] The purpose of the present invention is to solve the above-mentioned deficiencies in the prior art and to provide a pharmaceutical composition and use thereof.

[0007] The present invention is achieved through the following technical solutions: The invention provides a method for preparing a product for enhancing the efficacy of a drug for treating metabolic diseases, wherein the drug is an OB01 strain or a progeny strain or a subclone strain or a culture supernatant thereof or a preparation thereof, wherein the OB01 strain is a strain of Bifidobacterium bifidum ( Bifidobacterium bifidum )ibiome001, which is deposited in the Guangdong Provincial Microbiological Culture Collection Center, located at the 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou City, Guangdong Province, Institute of Microbiology, Guangdong Academy of Sciences, with a deposit date of May 17, 2022 and a deposit number of GDMCC No. 62473; the metabolic diseases are diabetes, obesity and obesity-related diseases, and metabolic syndrome; the obesity-related diseases include at least one of the following diseases: cardiovascular disease, hyperlipidemia, insulin resistance syndrome and fatty hepatitis.

[0008] The present invention also protects a pharmaceutical composition comprising: (a) OB01 strain or its progeny strain or subclone strain or its culture supernatant or preparation thereof, and (b) other drugs for the treatment of metabolic diseases; The OB01 strain is Bifidobacterium bifidum ( Bifidobacterium bifidum )ibiome001, which is deposited in the Guangdong Provincial Microbiological Culture Collection Center, located at the 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou City, Guangdong Province, Institute of Microbiology, Guangdong Academy of Sciences, with a deposit date of May 17, 2022 and a deposit number of GDMCC No. 62473; the metabolic diseases are diabetes, obesity and obesity-related diseases, and metabolic syndrome; the obesity-related diseases include at least one of the following diseases: cardiovascular disease, hyperlipidemia, insulin resistance syndrome and fatty hepatitis.

[0009] The above-mentioned pharmaceutical composition includes both the preparation of a compound preparation of the strain and other drugs, and the direct combination of the strain and other drugs. Among them, the total number of live bacteria contained in the live bacteria preparation is 10 6 -10 14 CFU; the medication cycle is based on the ability to achieve the desired effect, including but not limited to 1-3 times a day, 3-7 days a week, etc., and is also related to the concentration of the specific preparation.

[0010] The above preparation is the OB01 strain or its progeny strain or subclone strain or its culture supernatant, and pharmaceutically acceptable excipients.

[0011] There are many ways to classify excipients in preparations, which can be classified according to source, function and use, route of administration, etc. According to the source, they can be divided into natural products, semi-synthetic products and fully synthetic products. According to the function and use of excipients in preparations, there are 65 types of classifications, namely pH adjusters, chelating agents, inclusion agents, coating agents, protective agents, moisturizers, disintegrants, surfactants, virus inactivators, supplements, precipitants, film-forming materials, flavoring agents, freeze-drying excipients, carbon dioxide adsorbents, foaming agents, fragrances, preservatives, excipients, desiccant, curing agents, buffers, sustained-release materials, adhesives, flavoring agents, antioxidants, antioxidant enhancers, anti-adhesive agents, air replacement agents , condensing agent, ointment base material, gel material, polishing agent, propellant, solvent, softener, emulsifier, ointment base, soft capsule material, lubricant, wetting agent, penetration enhancer, osmotic pressure regulator, suppository base, sweetener, filler, pill core, stabilizer, adsorbent, absorbent, diluent, defoaming agent, flocculant, ethanol modifier, hard paste base, ink, thickener, solubilizer, plasticizer, adhesive, Chinese medicine preparation auxiliary materials, filter aid, solubilizer, suspending agent, colorant.

[0012] Preferably, the excipients include at least one of adjuvants, stabilizers or protective agents, antibacterial agents, excipients, solubilizers, flavoring agents, diluents, and buffers. Adjuvant: a substance mixed with one or more components that bind to a vaccine antigen to enhance [such as strengthen, accelerate, prolong and (or) possibly direct] its specific immune response and the clinical effect of the vaccine. Stabilizer or protective agent: a substance used to stabilize or protect the active ingredients of biological products to prevent them from degradation or loss of activity. Antibacterial agent: a substance used to inhibit microbial growth and prevent microbial contamination. Excipient: a substance used to shape the drug and act as a scaffold in freeze-dried products. Solubilizer: a substance used to increase the solubility of the drug. Flavoring agent: a substance used to improve the taste of oral drugs. Diluent, buffer: a solvent used to dissolve, dilute the product, and adjust the pH of the product, such as water for injection, sodium chloride injection, phosphate buffered physiological sodium chloride solution (PBS), etc.

[0013] Exemplary excipients include, but are not limited to, butylated hydroxytoluene (BHT), calcium carbonate, calcium phosphate (monohydrogen), calcium stearate, cross-linked carboxymethylcellulose, cross-linked polyvinyl pyrrolidone, citric acid, cross-linked povidone, cysteine, ethylcellulose, gelatin, hydroxypropyl cellulose, hydroxypropyl methylcellulose, lactose, magnesium stearate, maltitol, mannitol, methionine, methylcellulose, methylparaben, microcrystalline cellulose, polyethylene glycol, povidone, pregelatinized starch, propylparaben, retinyl palmitate, shellac, silicon dioxide, sodium carboxymethylcellulose, sodium citrate, sodium starch glycolate, sorbitol, starch (corn), stearic acid, stearic acid, sucrose, talc, titanium dioxide, vitamin A, vitamin E, vitamin C, and xylitol.

[0014] The pharmaceutical composition can be prepared in the form of an injection preparation and / or an oral preparation. The injection preparations are classified according to physical state, including liquid injections, powders for injection, and tablets for injection; and classified according to the injection part, including intradermal injections, subcutaneous injections, intramuscular injections, intravenous injections, and spinal cavity injections; preferably, the solvent of the injection preparation includes water for injection or physiological saline.

[0015] Formulations for oral use include tablets containing the active ingredient mixed with non-toxic pharmaceutically acceptable excipients. These excipients can be, for example, inert diluents or fillers (e.g., sucrose, sorbitol, sugar, mannitol, microcrystalline cellulose, starch including potato starch, calcium carbonate, sodium chloride, lactose, calcium phosphate, calcium sulfate or sodium phosphate); granulating agents and disintegrants (e.g., cellulose derivatives including microcrystalline cellulose, starch including potato starch, cross-linked carboxymethyl cellulose sodium, alginates or alginic acid); binders (e.g., sucrose, glucose, sorbitol, gum arabic, alginic acid, sodium alginate, gelatin, starch, pregelatinized starch, microcrystalline cellulose, magnesium aluminum silicate, sodium carboxymethyl cellulose, methylcellulose, hydroxypropyl methylcellulose, ethylcellulose, polyvinyl pyrrolidone or polyethylene glycol); and lubricants, glidants and anti-adherents (e.g., magnesium stearate, zinc stearate, stearic acid, silicon dioxide, hydrogenated vegetable oil or talc). Preparations for oral use may also be in the form of chewable tablets, or in the form of hard gelatin capsules in which the active ingredient is mixed with an inert solid diluent (e.g., potato starch, lactose, microcrystalline cellulose, calcium carbonate, calcium phosphate or kaolin), or in the form of soft gelatin capsules in which the active ingredient is mixed with water or an oil medium (e.g., peanut oil, liquid paraffin or olive oil). Powders, granules and pills can be prepared using the ingredients mentioned above under tablets or capsules in a conventional manner using, for example, a mixer, fluidized bed equipment or spray drying equipment.

[0016] Other pharmaceutically acceptable excipients for oral formulations include, but are not limited to, colorants, flavoring agents, plasticizers, humectants, and buffers. Formulations for oral use may also be in the form of chewable tablets, or in the form of hard gelatin capsules, wherein the active ingredient is mixed with an inert solid diluent (e.g., potato starch, lactose, microcrystalline cellulose, calcium carbonate, calcium phosphate, or kaolin), or in the form of soft gelatin capsules, wherein the active ingredient is mixed with water or an oil medium (e.g., peanut oil, liquid paraffin, or olive oil). Powders, granules, and pills may be prepared in a conventional manner using, for example, a mixer, fluidized bed apparatus, or spray drying apparatus using the ingredients mentioned above under tablets or capsules.

[0017] In some embodiments, administration comprises administering an agent described herein intramuscularly, intravenously (e.g., in the form of a sterile solution and in a solvent system suitable for intravenous use), intradermally, intraarterially, intraperitoneally, intralesionally, intracranially, intraarticularly, intraprostatically, intrapleurally, intratracheally, intranasally, intravitreally, intravaginally, intrarectally, topically, intratumorally, transperitoneally, subcutaneously, subconjunctivally, intracapsularly, transmucosally, intrapericardially, intraumbilically, intraocularly, orally (e.g., as a tablet, capsule, caplet, caplet-shaped tablet, or syrup), topically (e.g., in the form of a cream, gel, lotion, or ointment), topically, by inhalation, by injection, or by infusion (e.g., continuous infusion in the form of a cream or lipid composition, local perfusion, catheterization, lavage directly bathing target cells).

[0018] Preferably, the other metabolic disease therapeutic drugs include at least one of the following: insulin and insulin analogs, insulin secretagogues, insulin sensitizers, dipeptidyl peptidase-4 (DPP-4) inhibitors, sodium-glucose cotransporter 2 (SGLT-2) inhibitors, glucagon-like peptide-1 (GLP-1) receptor agonists, α-glucosidase inhibitors, appetite suppressants, metabolic regulators, statins, and fibrates. Furthermore, the insulin secretagogue is a sulfonylurea drug or a non-sulfonylurea drug; the insulin sensitizer is a biguanide drug or a thiazolidinedione drug.

[0019] Wherein, the insulin and insulin analogs are human insulin, glargine insulin, aspart insulin, lispro insulin, glulisine insulin, detemir insulin, degludec insulin or premixed insulin; the sulfonylurea drugs are glibenclamide, glipizide, gliclazide, gliquidone or glimepiride; the non-sulfonylurea drugs are repaglinide, nateglinide or mitiglinide; the biguanide drugs are metformin, metformin hydrochloride, metformin hydrochloride glibenclamide, pioglitazone metformin, metformin gliclazide, phenformin or buformin; the thiazolidinedione drugs are pioglitazone, rosiglitazone, troglitazone or ciglitazone; the DPP-4 inhibitors are sitagliptin, saxagliptin, vildagliptin, linagliptin, alogliptin, tenegliptin or gemagliptin; the S The GLT-2 inhibitor is dapagliflozin, canagliflozin, empagliflozin, rupagliflozin, ipragliflozin or togliflozin; the GLP-1 receptor agonist is semaglutide, dulaglutide, liraglutide, loxenatide, exenatide, semaglutide, albiglutide, belaglutide or lixisenatide; the α-glucosidase inhibitor is acarbose, voglibose or miglitol; the appetite suppressant is orlistat, fenfluramine, lorcaserin, cenikalan, bupropion, bupropion, diethylpropion or clofenindole; the statin is atorvastatin, simvastatin, rosuvastatin, fluvastatin, pravastatin, lovastatin, cerivastatin or rosuvastatin; the fibrate is fenofibrate, bezafibrate, gemfibrozil, clofibrate or ciprofibrate.

[0020] The present invention also protects the use of the above-mentioned pharmaceutical composition in preparing a drug for preventing or treating metabolic diseases, wherein the metabolic diseases are diabetes, obesity and obesity-related diseases, and metabolic syndrome.

[0021] In this specification, the term "diabetes" includes all forms of diabetes, which are characterized by metabolic disorders and abnormally high blood sugar (hyperglycemia) due to insufficient insulin hormone levels, as described above. The term thus includes type 1 diabetes (T1D), type 2 diabetes (T2D), gestational diabetes (GDM) and impaired glucose tolerance (IGT). Type 1 diabetes is caused by autoimmune damage or idiopathic causes, and is characterized by absolute destruction of pancreatic islet function. It mostly occurs in children and adolescents and must be treated with insulin to achieve satisfactory therapeutic effects, otherwise it will be life-threatening. Type 2 diabetes is a multifactorial syndrome characterized by abnormal carbohydrate / fat metabolism, usually including high blood sugar, high blood pressure and abnormal cholesterol. Type 2 diabetes is caused by the inability of insulin to work effectively (low content of binding to receptors), so not only fasting blood sugar should be checked, but also blood sugar 2 hours after a meal, and pancreatic islet function tests should be done in particular. There are two types of diabetes during pregnancy. One is that the patient has been diagnosed with diabetes before pregnancy, which is called "diabetes complicated with pregnancy"; the other is that the patient has normal glucose metabolism or potential glucose tolerance before pregnancy, and the diabetes only appears or is diagnosed during pregnancy, which is also called "gestational diabetes mellitus (GDM)". More than 80% of pregnant women with diabetes have GDM. Impaired glucose tolerance (IGT) is a pre-diabetic state of impaired glucose metabolism associated with insulin resistance and increased risk of cardiovascular disease. According to the standards of the World Health Organization and the American Diabetes Association, glucose intolerance is defined as a two-hour glucose level of 140-199 mg / dL (7.8 to 11.0 mmol) in a 75 g oral glucose tolerance test. When the patient has an immediately elevated glucose level after 2 hours, but is lower than the standard for type 2 diabetes, he or she is considered to be in an IGT state. Fasting glucose can be normal or slightly elevated. IGT can precede type 2 diabetes by many years. IGT is also a risk factor for death.

[0022] It should be noted that the above-mentioned pharmaceutical use for diabetes includes but is not limited to the treatment or prevention of type 1 diabetes (T1D), type 2 diabetes (T2D), gestational diabetes (GDM) and impaired glucose tolerance (IGT).

[0023] In this specification, the term "obesity" is associated with the body mass index (BMI). The body mass index (BMI), calculated as the weight in kilograms divided by the square of the height in meters, is the most commonly accepted calculation method for overweight and / or obesity. A BMI of more than 25 is considered overweight. A BMI of 30 or more is defined as obesity, a BMI of 35 or more is considered severe comorbidity obesity, and a BMI of 40 or more is considered morbid obesity.

[0024] As mentioned above, the term "obesity" as used herein includes overweight, obesity, comorbid obesity and morbid obesity. Therefore, the term "obesity" as used herein can be defined as a subject with a BMI greater than or equal to 25. In some embodiments, suitably, the BMI of an obese subject can be greater than or equal to 30, suitably 35, suitably 40.

[0025] Although the compositions of the present invention are particularly suitable for patients who suffer from both diabetes and obesity, the compositions of the present invention are also suitable for patients who suffer from diabetes but are not obese, and are also suitable for obese patients who have risk factors for diabetes but are not yet in a diabetic state. It is expected that obese people (but not diabetic) may limit the metabolic consequences of their obesity, i.e., the development of diabetes or at least insulin resistance.

[0026] In addition, the pharmaceutical composition for use in the present invention can be used to treat metabolic syndrome in mammals. Metabolic syndrome is a general term for medical conditions that increase the risk of cardiovascular disease and diabetes. Metabolic syndrome is also known as metabolic syndrome X, syndrome X, insulin resistance syndrome, Reaven's syndrome or CHAOS syndrome (Australia).

[0027] There is currently no single accepted definition of the metabolic syndrome. The World Health Organization criteria (1999) require the presence of diabetes mellitus, glucose intolerance, fasting glucose intolerance, or insulin resistance, and two of the following: Blood pressure: ≥140 / 90 mmHg; Dyslipidemia: triglyceride (TG): ≥1.695mmol / L, high-density lipoprotein cholesterol (HDL-C) ≤0.9mmol / L (male), ≤1.0mmol / L (female); Central obesity: waist:hip ratio > 0.90 (males), > 0.85 (females), and / or body mass index > 30 kg / m 2 .

[0028] Microalbuminuria: urinary albumin excretion rate ≥ 20 mg / min, or albumin: creatinine ratio ≥ 30 mg / g.

[0029] The European Insulin Resistance Study Group (1999) defines insulin resistance as a fasting insulin value greater than 25% of that in non-diabetic individuals, plus two or more of the following: Central obesity: waist circumference ≥94cm (male), ≥80cm (female); Dyslipidemia: TG ≥ 2.0 mmol / L, and / or HDL-C < 1.0 mg / dL, or being treated for dyslipidemia; Hypertension: blood pressure ≥140 / 90 mmHg, or taking antihypertensive drugs; Fasting blood glucose ≥6.1mmol / L.

[0030] The National Cholesterol Education Program (NCEP) Treatment Strategies for Adults III (2001) requires at least three of the following: Central obesity: waist circumference ≥102cm or 40 inches (male), ≥88cm or 36 inches (female); Dyslipidemia: TG ≥ 1.695mmol / L (150mg / dl); Dyslipidemia: HDL-C <40 mg / dL (male), <50 mg / dL (female); Blood pressure: ≥130 / 85 mmHg; Fasting blood glucose ≥6.1mmol / L (110mg / dl).

[0031] In addition, the obesity-related diseases include at least one of the following diseases: cardiovascular disease, hyperlipidemia, insulin resistance syndrome and steatohepatitis.

[0032] Examples of cardiovascular diseases that can be treated using the pharmaceutical composition according to the present invention include aneurysm, angina, atherosclerosis, cerebrovascular accident (stroke), cerebrovascular disease, congestive heart failure (CHF), coronary heart disease, myocardial infarction (heart attack), and peripheral vascular disease.

[0033] An aneurysm is a localized, blood-filled bulge (ballooning) in a blood vessel caused by a diseased or weakened blood vessel wall. Aneurysms most commonly occur in the arteries at the base of the brain (cerebral arterial circle) and in the aorta (the main artery leaving the heart, a so-called aortic aneurysm). As an aneurysm grows, the risk of rupture increases, which can lead to severe bleeding or other complications, including sudden death.

[0034] Angina pectoris (commonly called angina) is severe chest pain due to myocardial ischemia (lack of blood and thus oxygen supply), usually caused by blockage or spasm of the coronary arteries (blood vessels of the heart). Coronary heart disease (the main cause of angina) is caused by atherosclerosis of the heart's arteries.

[0035] Atherosclerosis is a condition in which the walls of arteries thicken due to the accumulation of fatty substances, such as cholesterol. It is a syndrome that affects the arterial blood vessels and is a chronic inflammatory response of the arterial walls, largely due to the accumulation of white blood cells called macrophages, and is promoted by low-density (especially small-particle) lipoproteins (plasma proteins that carry cholesterol and triglycerides) and the inability of functional high-density lipoproteins (HDL) to properly remove fat and cholesterol from macrophages. Atherosclerosis is often referred to as hardening of the arteries or "furring" and is usually caused by the formation of multiple plaques within the arteries.

[0036] A stroke is a rapid loss of brain function due to a disruption in the blood supply to the brain. This may be due to ischemia (lack of blood supply) caused by a blood clot or embolism, or due to bleeding. As a result, the affected area of ​​the brain cannot function, resulting in loss of movement of one or more limbs on one side of the body, loss of ability to understand or describe speech, or loss of vision on one side of the body and ultimately death.

[0037] Cerebrovascular disease is a group of brain dysfunctions associated with vascular diseases that supply the brain. Hypertension is the most important cause of damage to the endothelium of the inner membrane of the blood vessels in contact with the underlying collagen, where platelets accumulate in the underlying collagen and initiate a repair process that is not always complete and perfect. Persistent hypertension permanently changes the structure of the blood vessels, making them narrow, stiff, deformed, and uneven, making them more susceptible to blood pressure fluctuations. A decrease in blood pressure during sleep can lead to a significant decrease in blood flow in the narrowed blood vessels, leading to morning ischemic strokes, while a sudden increase in blood pressure can cause blood vessel ruptures, leading to intracranial hemorrhage during daytime excitement. Mainly elderly people, diabetics, smokers, or people with ischemic heart disease suffer from cerebrovascular disease. All diseases related to arterial dysfunction can be classified in diseases called large vessel diseases. This is an oversimplified study, which allows fat deposits or thrombi to block arteries. The results of cerebrovascular disease may include stroke, or even sometimes hemorrhagic stroke. In the process of cerebrovascular accidents, ischemia or other vascular dysfunctions may affect one.

[0038] Heart failure is a general term for a physiological state in which cardiac output is insufficient for the body's needs. This can occur in the setting of low cardiac output (often referred to as "congestive heart failure"). Common causes of heart failure include myocardial infarction and other forms of ischemic heart disease, hypertension, valvular heart disease, and cardiomyopathy.

[0039] Coronary heart disease (or coronary artery heart disease) is the inability of the coronary artery circulation to supply adequate circulation to the heart muscle and surrounding tissues. It is often equated with atherosclerotic coronary artery heart disease, but coronary artery disease can be attributed to other causes, such as coronary vasospasm. Stenosis may be caused by spasm.

[0040] Myocardial infarction (commonly called a heart attack) occurs when the blood supply to part of the heart is blocked, causing certain heart cells to die. This is most often attributed to blockage (occlusion) of a coronary artery following the rupture of a vulnerable atherosclerotic plaque (an unstable accumulation of lipids (such as cholesterol) and white blood cells (especially macrophages) in the artery wall). The resulting ischemia (restricted blood supply) and lack of oxygen, if left untreated long enough, can lead to myocardial damage and / or necrosis (infarction).

[0041] Peripheral vascular disease (PVD), also known as peripheral arterial disease (PAD) or peripheral arterial occlusive disease (PAOD), includes all diseases caused by occlusion of the large arteries in the arms and legs. PVD can be attributed to atherosclerosis, inflammatory processes leading to stenosis, embolism, or thrombosis, which results in acute or chronic ischemia (lack of blood supply), usually in the legs.

[0042] It is contemplated within the scope of the present invention that the embodiments of the present invention may be combined, such that any combination of features described herein is also included within the scope of the present invention. In particular, it is contemplated within the scope of the present invention that any therapeutic effects of the composition may be simultaneously manifested.

[0043] Preferably, the drug is used for at least one of the following purposes: preventing or treating diabetes caused by obesity; preventing or treating type II diabetes; reducing the weight of mammals; reducing fasting blood glucose in mammals; improving impaired oral glucose tolerance in mammals; improving insulin resistance in mammals; reducing the area under the insulin tolerance curve; improving the cholesterol content in the blood of mammals; reducing the glycated hemoglobin content in mammals.

[0044] It is worth noting that when measuring the effect of combined drug use, the Q value method, also known as the Kim Jong-kyun Q value method, is often used. It is a statistical method used to evaluate whether two or more drugs, treatment methods, etc. produce a synergistic effect when used in combination. This method determines whether there is a synergistic effect by comparing the difference in effect when the drug is used in combination with that when it is used alone. Suppose the effects of drug A and drug B when used alone are E and A and E B , the effect when used together is E AB , then the Q value can be calculated by the following formula: Q = E AB / (EA + EB − EA × EB)When the Q value is greater than 1, it means that the effect of the combined drug is greater than the simple sum of the effects of the individual drugs, that is, there is a synergistic effect; when the Q value is equal to 1, it means that the effect of the combined drug is equal to the simple sum of the effects of the individual drugs, that is, it shows an additive effect; when the Q value is less than 1, it means that the effect of the combined drug is less than the simple sum of the effects of the individual drugs, and there may be an antagonistic effect.

[0045] The beneficial effects of the present invention are: When OB01 is used in combination with metformin, semaglutide, sitagliptin, and dapagliflozin, blood sugar, blood lipids, and obesity are improved to varying degrees, achieving a synergistic effect, which can improve efficacy, reduce drug dosage, regulate the balance of intestinal flora, reduce adverse reactions caused by commonly used drugs, and help reduce drug resistance to commonly used drugs by regulating the body's metabolic process. The combination of OB01 and common positive drugs provides patients with a safer and more effective treatment plan, which has important clinical significance and application value. BRIEF DESCRIPTION OF THE DRAWINGS

[0046] Figure 1 The figure shows the effect of the combined use of OB01 and metformin on the insulin tolerance of ob / ob mice in Example 1 (A is insulin tolerance, B is the area under the insulin tolerance curve).

[0047] Figure 2 The figure shows the effect of the combined use of OB01 and semaglutide on the insulin tolerance of ob / ob mice in Example 2 (A is insulin tolerance, B is the area under the insulin tolerance curve).

[0048] Figure 3 The figure shows the effect of the combined use of OB01 and semaglutide on the body weight of DIO mice in Example 3 (A is the body weight curve from 0 to 20 weeks, and B is the body weight value at 20 weeks).

[0049] Figure 4 The effect of the combined use of OB01 and semaglutide in Example 3 on blood lipids in DIO mice is shown.

[0050] Figure 5 The figure shows the effect of the combined use of OB01 and semaglutide in Example 3 on fasting blood glucose (A) and glycated hemoglobin (B) in DIO mice.

[0051] Figure 6 The figure shows the effect of the combined use of OB01 and semaglutide on the oral glucose tolerance of DIO mice in Example 3 (A is oral glucose tolerance, B is the area under the oral glucose tolerance curve).

[0052] Figure 7The effect of the combined use of OB01 and sitagliptin on the insulin tolerance of DIO mice in Example 4 is shown (A is insulin tolerance, B is the area under the insulin tolerance curve). DETAILED DESCRIPTION

[0053] For a better understanding of the present invention, the present invention is further described below in conjunction with embodiments and drawings. The following embodiments are merely for illustration of the present invention rather than for limitation thereof. Example 1 Effects of OB01 combined with metformin on ob / ob mice

[0054] 1. Experimental methods: Forty 8-week-old ob / ob mice (bred by Han Microbiology) were randomly divided into (1) control group (Model), (2) OB01 group, (3) metformin group (Met), and (4) OB01 and metformin combined group (OB01+Met), with 10 mice in each group. The mice in each group were fed with normal feed while receiving medication until the end of the experiment.

[0055] (1) Control group: intragastrically administered 0.2 mL 0.9% saline / animal / day; (2) OB01 group: Gavage OB01 (i.e., Bifidobacterium bifidum ibiome001, deposited on May 17, 2022 in Guangdong Microbial Culture Collection Center, address: 5th Floor, Building 59, No. 100, Xianlie Middle Road, Guangzhou City, Guangdong Province, Institute of Microbiology, Guangdong Academy of Sciences, with the deposit number of GDMCC No. 62473). According to the number of viable bacteria in each batch of freeze-dried bacterial powder after OB01 was prepared into freeze-dried bacterial powder, the number of viable bacteria per gavage was calculated. 9 The number of grams of bacterial powder required for CFU / each was resuspended in 0.2 mL of 0.9% saline once a day; (3) Metformin group (Met): Metformin 100 mg / kg (equivalent to 0.5 g / day for humans) was administered orally, dissolved in normal saline, 0.2 mL per mouse, once a day; (4) OB01 and metformin combined group (OB01+Met): intragastric administration for 10 9 CFU / OB01 and 100 mg / kg metformin were dissolved in normal saline and administered intragastrically with 0.2 mL per mouse once a day.

[0056] Insulin tolerance (ITT) was tested at the 13th week of administration: the mice were fasted for 4 hours, blood was collected from the tip of the tail (0 h) to measure blood glucose, and 0.75 IU / kg insulin solution was injected subcutaneously. Then, blood glucose was measured at the tip of the tail at 30 min, 60 min, and 120 min, and the blood glucose change curve over time was plotted, and the area under the curve was calculated.

[0057] 2. Experimental results: like Figure 1 As shown in the figure, compared with the control group, oral administration of OB01, metformin, and OB01 combined with metformin can significantly reduce the area under the insulin tolerance curve of ob / ob mice, and oral administration of OB01 is more effective than oral administration of metformin. At the same time, the effect of OB01 combined with metformin is significantly better than that of oral administration of OB01 or metformin alone (the AUCof ITT decrease rate is calculated using the Q value method, Q=1.07). This shows that oral administration of OB01 alone is expected to replace metformin and avoid the side effects of metformin such as nausea, vomiting, diarrhea, abdominal pain, bloating, indigestion, fatigue, etc.; when OB01 is combined with metformin, the dosage of metformin can be reduced while ensuring the same efficacy, and OB01 can be used to regulate the patient's intestines, thereby reducing the possibility of side effects and drug resistance. Example 2 Effects of OB01 combined with semaglutide on ob / ob mice

[0058] 1. Experimental methods: Forty 8-week-old ob / ob mice (bred by Hanwei Bio) were randomly divided into (1) control group (Model), (2) OB01 group, (3) semaglutide group (Sema), and (4) OB01 and semaglutide combined group (OB01+Sema), with 10 mice in each group. The mice in each group were fed with normal feed at the same time as the drug administration until the end of the experiment.

[0059] (1) Control group: intragastrically administered 0.2 mL 0.9% saline / animal / day + subcutaneously injected 0.1 mL 0.9% saline / animal / week; (2) OB01 group: Gavage OB01 (i.e., Bifidobacterium bifidum ibiome001, deposited on May 17, 2022 in Guangdong Microbial Culture Collection Center, address: 5th Floor, Building 59, No. 100, Xianlie Middle Road, Guangzhou City, Guangdong Province, Institute of Microbiology, Guangdong Academy of Sciences, with the deposit number of GDMCC No. 62473). According to the number of viable bacteria in each batch of freeze-dried bacterial powder after OB01 was prepared into freeze-dried bacterial powder, the number of viable bacteria per gavage was calculated. 9 The number of grams of bacterial powder required for CFU / mouse was resuspended in 0.2 mL of 0.9% saline once a day; in addition, 0.1 mL of 0.9% saline was injected subcutaneously / mouse / week; (3) Semaglutide group (Sema): Semaglutide was injected subcutaneously. Each mouse was given semaglutide according to body weight (12.8 μg / kg / week from the first to the fifth week, 25.6 μg / kg / week from the sixth to the fifteenth week). Semaglutide was diluted with normal saline and 0.1 mL was injected subcutaneously to each mouse once a week. In addition, 0.2 mL of 0.9% normal saline was gavaged per mouse per day. (4) OB01 and semaglutide combined group (OB01+Sema): intragastrically administered for 10 9 CFU / mouse / day OB01 (resuspended in 0.2 mL 0.9% saline) and subcutaneous injection of semaglutide (12.8 μg / kg / week from the first to the fifth week, 25.6 μg / kg / week from the sixth to the fifteenth week, diluted with saline, 0.1 mL per mouse subcutaneously, once a week).

[0060] Insulin tolerance (ITT) was tested at the 15th week of administration: the mice were fasted for 4 hours, blood was collected from the tip of the tail (0 h) to measure blood glucose, and 0.75 IU / kg insulin solution was injected subcutaneously. Then, blood glucose was measured at the tip of the tail at 30 min, 60 min, and 120 min, and the blood glucose change curve over time was plotted, and the area under the curve was calculated.

[0061] 2. Experimental results: like Figure 2 As shown in the figure, compared with the control group, oral administration of OB01, semaglutide, and the combination of OB01 and semaglutide can significantly reduce the area under the insulin tolerance curve of ob / ob mice. At the same time, the combined effect of OB01 and semaglutide is significantly better than that of oral administration of OB01 or semaglutide alone (the AUC of ITT decrease rate is calculated using the Q value method, Q=1.19). Therefore, when OB01 and semaglutide are selected for combined use, the dosage of patients taking semaglutide for obesity and metabolic diseases caused by leptin deficiency can be reduced while ensuring the same drug efficacy. OB01 can be used to regulate the intestines of patients, thereby reducing the side effects of semaglutide and reducing the possibility of drug resistance. Example 3 Effects of OB01 combined with semaglutide on DIO mice

[0062] 1. Experimental methods: Forty-five 8-week-old C57BL / 6J mice (purchased from Jiangsu Jicui Pharmaceutical Biotechnology Co., Ltd.) were randomly divided into (1) negative control group (control), (2) control group (Model), (3) OB01 group, (4) semaglutide group (Sema), and (5) OB01 and semaglutide combined treatment group (OB01+Sema), with 5-10 mice in each group. Except for the negative control group (control), which was fed with ordinary maintenance feed, the mice in the other groups were fed with high-fat feed (60 kcal%Fat, research diet, D12492) at the same time as the drug administration until the end of the experiment.

[0063] (1) Negative control group (control): intragastric administration of 0.2 mL 0.9% saline / animal / day + subcutaneous injection of 0.1 mL 0.9% saline / animal / week; (2) Control group (Model): intragastrically administered 0.2 mL 0.9% saline / animal / day + subcutaneously injected 0.1 mL 0.9% saline / animal / week; (3) OB01 group: Gavage OB01 (i.e., Bifidobacterium bifidum ibiome001, deposited on May 17, 2022 in Guangdong Microbial Culture Collection Center, address: 5th Floor, Building 59, No. 100, Xianlie Middle Road, Guangzhou City, Guangdong Province, Institute of Microbiology, Guangdong Academy of Sciences, with the deposit number of GDMCC No. 62473). According to the number of viable bacteria in each batch of freeze-dried bacterial powder after OB01 was prepared into freeze-dried bacterial powder, the number of viable bacteria per gavage was calculated. 8 The number of grams of bacterial powder required for CFU / mouse was resuspended in 0.2 mL of 0.9% saline once a day; in addition, 0.1 mL of 0.9% saline was subcutaneously injected / mouse / week; (4) Semaglutide group (Sema): Subcutaneous injection of semaglutide, each mouse was given 12.8 μg / kg semaglutide according to body weight, diluted with normal saline, and each mouse was subcutaneously injected with 0.1 mL, once a week. In addition, 0.2 mL 0.9% normal saline was gavaged per mouse per day; (5) OB01 and semaglutide combined group (OB01+Sema): intragastrically administered for 10 8 CFU / mouse / day OB01 (resuspended in 0.2 mL 0.9% saline) and subcutaneous injection of 12.8 μg / kg / week semaglutide (diluted with saline, 0.1 mL per mouse subcutaneously, once a week).

[0064] The following experiments were conducted respectively: a. Monitor body weight weekly; b. Fasting blood glucose was tested at the 12th week of administration (fasting for 6 hours); c. Oral glucose tolerance (OGTT) test at the 14th week of administration: mice were fasted for 6 hours and blood was collected from the tail tip (0 h) to measure blood glucose, and 2 g / kg glucose solution was gavaged intragastrically. Then, blood was collected from the tail tip at 30 min, 60 min, and 120 min to measure blood glucose. The blood glucose change curve over time was plotted, and the area under the curve was calculated. d. After 20 weeks of administration, the mice were fasted for 12 h, their eyeballs were removed and blood was collected into heparin tubes. The blood was centrifuged at 4000 rpm for 10 min at room temperature, and the supernatant plasma was transferred to a new centrifuge tube and stored at -80°C for later use. The glycated hemoglobin content and low-density lipoprotein cholesterol content in the blood were detected using a kit.

[0065] 2. Experimental results (1) Weight like Figure 3 As shown, compared with the control group (Model), starting from the seventh week of administration, oral administration of OB01, semaglutide, and the combination of OB01 and semaglutide were able to significantly reduce the body weight of DIO mice, and the effect lasted until the end of administration; at the same time, the combination of OB01 and semaglutide was significantly better than oral administration of OB01 or semaglutide alone, and the effect was almost the same as that of the negative control group (control) (the weight loss rate was calculated using the Q value method, Q=1.05).

[0066] (2) Blood lipids like Figure 4 As shown, compared with the control group (Model), oral administration of OB01, semaglutide, and the combination of OB01 and semaglutide can significantly reduce the plasma low-density lipoprotein (LDL-C) content of DIO mice; at the same time, the combination of OB01 and semaglutide is significantly better than oral administration of OB01 or semaglutide alone (the low-density lipoprotein decrease rate is calculated using the Q value method, Q=1.10).

[0067] (3) Blood sugar (i) Fasting blood glucose and glycated hemoglobin levels like Figure 5 As shown, compared with the control group (Model), oral administration of OB01, semaglutide, and the combination of OB01 and semaglutide can significantly reduce the fasting blood glucose and glycosylated hemoglobin levels of DIO mice; at the same time, the combination of OB01 and semaglutide is significantly better than oral administration of OB01 or semaglutide alone in controlling fasting blood glucose and glycosylated hemoglobin levels, and the effect is not significantly different from the negative control group (control). Among them, the glycosylated hemoglobin content can reflect the long-term control effect of blood glucose. (The decrease rates calculated by the Q value method are Q=1.01 and Q=1.12 respectively) (ii) Oral glucose tolerance like Figure 6 As shown, compared with the control group (Model), oral administration of OB01, semaglutide, and the combination of OB01 and semaglutide can significantly reduce the area under the oral glucose tolerance curve of DIO mice; at the same time, the combination of OB01 and semaglutide is significantly better than oral administration of OB01 or semaglutide alone, and the effect is better than the negative control group (control) (the AUC of OGTT decrease rate was calculated using the Q value method, Q=1.02).

[0068] Therefore, when OB01 is used in combination with semaglutide, the dosage of semaglutide for patients taking semaglutide for obesity and metabolic diseases caused by a high-fat diet can be reduced while ensuring the same efficacy. OB01 can be used to regulate the patient's intestines, thereby reducing the side effects of semaglutide and reducing the possibility of drug resistance. Example 4 Effects of OB01 combined with sitagliptin on DIO mice

[0069] 1. Experimental methods: Twenty-four C57BL / 6J mice (DIO mice) (purchased from Beijing Biotechnology Co., Ltd.) with 60% high-fat diet-induced obesity (weight range: 37.5g-43.1g) were randomly divided into (1) control group (Model), (2) OB01 group, (3) sitagliptin group (Stp) and (4) OB01 and sitagliptin combination group (OB01+Stp), with 6 mice in each group. The mice in each group continued to be fed with a high-fat diet (60 kcal% Fat, research diet, 12492) until the end of the experiment.

[0070] (1) Control group (Model): intragastrically administered with 0.2 mL 0.9% saline / rat / day; (2) OB01 group: Gavage OB01 (i.e., Bifidobacterium bifidum ibiome001, deposited on May 17, 2022 in Guangdong Microbial Culture Collection Center, address: 5th Floor, Building 59, No. 100, Xianlie Middle Road, Guangzhou City, Guangdong Province, Institute of Microbiology, Guangdong Academy of Sciences, with the deposit number of GDMCC No. 62473). According to the number of viable bacteria in each batch of freeze-dried bacterial powder after OB01 was prepared into freeze-dried bacterial powder, the number of viable bacteria per gavage was calculated. 8 The number of grams of bacterial powder required for CFU / each was resuspended in 0.2 mL of 0.9% saline once a day; (3) Sitagliptin group (Stp): Sitagliptin 20 mg / kg was administered orally, dissolved in normal saline, 0.2 mL per mouse, once a day.

[0071] (4) OB01 and sitagliptin combined group (OB01+Stp): intragastric administration for 10 8 CFU / OB01 and 20 mg / kg sitagliptin were dissolved in normal saline and gavaged 0.2 mL per mouse once a day.

[0072] Insulin tolerance (ITT) was tested at the 8th week of administration: the mice were fasted for 4 hours, blood was collected from the tip of the tail (0 h) to measure blood glucose, and 0.75 IU / kg insulin solution was injected subcutaneously. Then, blood glucose was measured at the tip of the tail at 30 min, 60 min, and 120 min, and the blood glucose change curve over time was plotted, and the area under the curve was calculated.

[0073] 2. Experimental results: like Figure 7 As shown in the figure, compared with the control group, oral administration of OB01, sitagliptin and the combination of OB01 and sitagliptin can significantly reduce the area under the insulin tolerance curve of DIO mice, and there is no significant difference between oral administration of OB01 and oral administration of sitagliptin. At the same time, the combined use of OB01 and sitagliptin is better than oral administration of OB01 or sitagliptin alone. In particular, at 30 minutes of the ITT experiment, the combined use of OB01 and sitagliptin is significantly better than oral administration of OB01 or sitagliptin alone (the decrease rate at 30 minutes of the ITT experiment was calculated using the Q value method, Q=1.22). This shows that oral administration of OB01 alone is expected to replace sitagliptin and avoid the adverse reactions that may be caused by sitagliptin, such as neurogenic inflammation, increased blood pressure, and immune response. When OB01 and sitagliptin are used in combination, the dosage of sitagliptin can be reduced while ensuring the same efficacy, and OB01 can be used to regulate the patient's intestines, thereby reducing the possibility of adverse reactions and drug resistance.

[0074] The above-described embodiments are merely descriptions of preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should all fall within the protection scope determined by the claims of the present invention.

Claims

1. Use of a drug in the preparation of a product for enhancing the efficacy of a drug for treating metabolic diseases, characterized in that: The drug is the OB01 strain or its progeny strain or subclone strain or its culture supernatant or its preparation, and the OB01 strain is Bifidobacterium bifidum ( Bifidobacterium bifidum )ibiome001, which is deposited in Guangdong Microbiological Culture Collection Center, located at 5th Floor, Building 59, No. 100, Xianlie Middle Road, Guangzhou City, Guangdong Province, Institute of Microbiology, Guangdong Academy of Sciences, deposit date May 17, 2022, deposit number GDMCC No. 62473; The metabolic diseases are diabetes, obesity and obesity-related diseases, and metabolic syndrome; the obesity-related diseases include at least one of the following diseases: cardiovascular disease, hyperlipidemia, insulin resistance syndrome, and fatty liver disease.

2. The use according to claim 1, characterized in that: The preparation is OB01 strain or its progeny strain or subclone strain or its culture supernatant, and pharmaceutically acceptable adjuvants.

3. A pharmaceutical composition, characterized in that Include: (a) OB01 strain or its progeny strain or subclone strain or its culture supernatant or preparation thereof, and (b) other drugs for the treatment of metabolic diseases; The OB01 strain is Bifidobacterium bifidum ( Bifidobacterium bifidum )ibiome001, which is deposited in Guangdong Microbiological Culture Collection Center, located at 5th Floor, Building 59, No. 100, Xianlie Middle Road, Guangzhou City, Guangdong Province, Institute of Microbiology, Guangdong Academy of Sciences, deposit date May 17, 2022, deposit number GDMCC No. 62473; The metabolic diseases are diabetes, obesity and obesity-related diseases, and metabolic syndrome; the obesity-related diseases include at least one of the following diseases: cardiovascular disease, hyperlipidemia, insulin resistance syndrome, and fatty liver disease.

4. The pharmaceutical composition according to claim 3, characterized in that: The preparation described in (a) is the OB01 strain or its progeny strain or subclone strain or its culture supernatant, and pharmaceutically acceptable excipients.

5. The pharmaceutical composition according to claim 3, characterized in that The other metabolic disease therapeutic drugs include at least one of the following: insulin and insulin analogs, insulin secretagogues, insulin sensitizers, dipeptidyl peptidase-4 (DPP-4) inhibitors, sodium-glucose co-transporter 2 (SGLT-2) inhibitors, glucagon-like peptide-1 (GLP-1) receptor agonists, α-glucosidase inhibitors, appetite suppressants, metabolic regulators, statins, and fibrates.

6. The pharmaceutical composition according to claim 5, characterized in that: The insulin secretagogue is a sulfonylurea drug or a non-sulfonylurea drug; The insulin sensitizer is a biguanide drug or a thiazolidinedione drug.

7. The pharmaceutical composition according to claim 6, characterized in that: The insulin and insulin analogs are human insulin, insulin glargine, insulin aspart, insulin lispro, insulin glulisine, insulin detemir, insulin degludec or premixed insulin; The sulfonylurea drug is glibenclamide, glipizide, gliclazide, gliquidone or glimepiride; The non-sulfonylurea drug is repaglinide, nateglinide or mitiglinide; The biguanide drug is metformin, metformin hydrochloride, metformin hydrochloride glibenclamide, pioglitazone metformin, metformin gliclazide, phenformin or buformin; The thiazolidinedione drug is pioglitazone, rosiglitazone, troglitazone or ciglitazone; The DPP-4 inhibitor is sitagliptin, saxagliptin, vildagliptin, linagliptin, alogliptin, tenegliptin or gemagliptin; The SGLT-2 inhibitor is dapagliflozin, canagliflozin, empagliflozin, rupagliflozin, empagliflozin or topagliflozin; The GLP-1 receptor agonist is semaglutide, dulaglutide, liraglutide, loxenatide, exenatide, semaglutide, albiglutide, belaglutide or lixisenatide; The α-glucosidase inhibitor is acarbose, voglibose or miglitol; The appetite suppressant is orlistat, fenfluramine, lorcaserin, cenikalan, bupropion, bupropion, diethylpropion or clofenin; The statin is atorvastatin, simvastatin, rosuvastatin, fluvastatin, pravastatin, lovastatin, cerivastatin or rosuvastatin; The fibrate drug is fenofibrate, bezafibrate, gemfibrozil, clofibrate or ciprofibrate.

8. The pharmaceutical composition according to claim 3, characterized in that: The pharmaceutical composition is an injection preparation and / or an oral preparation.

9. Use of the pharmaceutical composition according to any one of claims 3 to 7 in the preparation of a medicament for preventing or treating a metabolic disease, characterized in that: The metabolic diseases are diabetes, obesity and obesity-related diseases, and metabolic syndrome; the obesity-related diseases include at least one of the following diseases: cardiovascular disease, hyperlipidemia, insulin resistance syndrome, and fatty liver disease.

10. The use according to claim 9, characterized in that The drug is used for at least one of the following purposes: Prevent or treat obesity-induced diabetes; Prevent or treat type 2 diabetes; Reducing the weight of mammals; Lowering fasting blood sugar in mammals; Improve impaired oral glucose tolerance in mammals; Improve insulin resistance in mammals; Reduced area under the insulin tolerance curve; Improve the cholesterol content in the blood of mammals; Lowering the glycated hemoglobin content in mammals.

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