Application of polyketides in the treatment of metabolic disorder-related diseases

Abstract

The present invention relates to the use of polyketide compounds in treating diseases related to metabolic disorders, and specifically to the use of the following compound of formula I, its tautomers, pharmaceutically acceptable salts or esters in preparing drugs for preventing or treating diseases related to lipid and / or sugar metabolism disorders:

Description

Technical Field

[0001] The present invention relates to the field of medicine, and in particular to the use of polyketide compounds in treating diseases related to metabolic disorders (such as sugar and / or lipid metabolism disorders). Background Art

[0002] Metabolic disorder refers to a pathological state in which the body's glucose, lipid and / or protein metabolism is abnormal, resulting in an imbalance between supply and demand. With changes in dietary structure and lifestyle, obesity, insulin resistance, hypercholesterolemia, non-alcoholic fatty liver disease, etc. caused by metabolic disorders have become a global epidemic. The important clinical manifestations of this type of disease are increased weight, increased body fat percentage, increased blood sugar after meals but difficulty in regulating blood sugar with insulin, increased fat in the liver and abnormal liver hypertrophy. This will further cause or aggravate the risk of liver and cardiovascular and cerebrovascular diseases.

[0003] Finding drugs with novel structures that have good therapeutic effects on the above-mentioned diseases has become an urgent problem to be solved in this field. Summary of the invention

[0004] The high-fat diet mouse model is one of the model animals induced by the Western diet. It is mainly based on high-calorie, high-fat food to induce glucose and lipid metabolism disorders in mice, and is used to replicate the production of obesity, insulin resistance, hypercholesterolemia, and non-alcoholic fatty liver. C57BL mice are a special strain of mice that are sensitive to a high-fat diet, and male mice are more sensitive than female mice. The animal model is mainly manifested by increased body weight, increased body fat percentage, increased liver wet weight, increased total triglycerides in the liver, increased total cholesterol and low-density lipoprotein in serum, increased fasting blood sugar, increased blood sugar curves in glucose tolerance tests, and the appearance of a large number of fat vacuoles in liver tissue sections. Through a series of inspection methods, it can be determined whether the model is successful and can be used to observe the effects of target compounds on obesity, diabetes, hypercholesterolemia, and non-alcoholic fatty liver.

[0005] The inventors cultured the sea mud fungus MMF4 in a rice culture medium and isolated compound J2D (the structure of which is described herein). The compound was found to significantly reduce the body weight, body fat percentage, liver wet weight, and total triglyceride content in liver tissue and serum of the model mice through a high-fat mouse model, and also improve insulin resistance. At the same time, the liver sections of the model mice showed that the fat vacuoles in the administration group were significantly reduced. Therefore, the compound is very beneficial for the treatment of metabolic disorders (especially lipid metabolism disorders) and related diseases, and has important clinical application value.

[0006] One aspect of the present invention provides the use of the following compound of formula I, its tautomer, pharmaceutically acceptable salt or ester, or a pharmaceutical composition containing any of the above ingredients in the preparation of a drug for preventing or treating diseases related to lipid and / or sugar metabolism disorders:

[0007]

[0008] Formula I

[0009] Wherein, R1 and R2 are each independently selected from a linear or branched C 2-10 Alkyl, C 2-10 Alkenyl and C 2-10 Alkynyl.

[0010] In some embodiments, R1 and R2 in the compound of formula I are each independently selected from a linear or branched C 4-6 Alkyl, C 4-6 Alkenyl and C 4-6 Alkynyl.

[0011] In some embodiments, in the compound of formula I, R1 and R2 are each independently selected from a linear C 4-6 In some preferred embodiments, the C 4-6 Alkenyl groups contain 1 or 2 carbon-carbon double bonds.

[0012] In some embodiments, R1 and R2 in the compound of formula I are each independently selected from and .

[0013] In some embodiments, the compound of formula I is .

[0014] In some embodiments, the lipid and / or sugar metabolism disorder-related disease is selected from one or more of metabolic syndrome, obesity, diabetes (eg, type II diabetes), prediabetes, insulin resistance, liver disease, and cardiovascular disease.

[0015] In some embodiments, the liver disease is fatty liver (eg, non-alcoholic fatty liver disease) or cirrhosis.

[0016] In some embodiments, the cardiovascular disease is selected from hypertension, hyperlipidemia (eg, hypercholesterolemia), coronary heart disease, myocardial infarction, and atherosclerosis.

[0017] In another aspect, the present invention provides a method for preventing or treating diseases related to lipid and / or glucose metabolism disorders, comprising the step of administering to a subject in need thereof an effective amount of the following compound of formula I, its tautomer, pharmaceutically acceptable salt or ester, or a pharmaceutical composition containing any of the above ingredients:

[0018]

[0019] Formula I

[0020] Wherein, R1 and R2 are each independently selected from a linear or branched C 2-10 Alkyl, C 2-10 Alkenyl and C 2-10 Alkynyl.

[0021] In some embodiments, R1 and R2 in the compound of formula I are each independently selected from a linear or branched C 4-6 Alkyl, C 4-6 Alkenyl and C 4-6 Alkynyl.

[0022] In some embodiments, in the compound of formula I, R1 and R2 are each independently selected from a linear C 4-6 In some preferred embodiments, the C 4-6 Alkenyl groups contain 1 or 2 carbon-carbon double bonds.

[0023] In some embodiments, R1 and R2 in the compound of formula I are each independently selected from and .

[0024] In some embodiments, the compound of formula I is .

[0025] In some embodiments, the lipid and / or sugar metabolism disorder is selected from one or more of metabolic syndrome, obesity, diabetes (eg, type II diabetes), prediabetes, insulin resistance, liver disease, and cardiovascular disease.

[0026] In some embodiments, the liver disease is fatty liver (eg, non-alcoholic fatty liver disease) or cirrhosis.

[0027] In some embodiments, the cardiovascular disease is selected from hypertension, hyperlipidemia (eg, hypercholesterolemia), coronary heart disease, myocardial infarction, and atherosclerosis.

[0028] In some embodiments, the pharmaceutical composition of the present invention can be formulated into any dosage form known in the medical field, for example, tablets, pills, suspensions, emulsions, solutions, gels, capsules, powders, granules, elixirs, lozenges, suppositories, injections (including injections, sterile powders for injection and concentrated solutions for injection), inhalants, sprays, etc. The preferred dosage form depends on the intended mode of administration and therapeutic use. The pharmaceutical composition of the present invention should be sterile and stable under production and storage conditions. A preferred dosage form is an injection. Such an injection can be a sterile injection solution. For example, a sterile injection solution can be prepared by the following method: incorporating a necessary dose of a compound of the present invention, a pharmaceutically acceptable salt or ester thereof, a stereoisomer thereof, or a solvate thereof in an appropriate solvent, and optionally, incorporating other desired ingredients (including but not limited to, pH adjusters, surfactants, ionic strength enhancers, isotonic agents, preservatives, diluents, or any combination thereof), followed by filtration and sterilization. In addition, sterile injection solutions can be prepared as sterile lyophilized powders (e.g., by vacuum drying or freeze drying) for storage and use. Such sterile lyophilized powders can be dispersed in a suitable carrier, such as sterile pyrogen-free water, before use.

[0029] Furthermore, the compound of the present invention, its tautomer, pharmaceutically acceptable salt or ester may be present in a pharmaceutical composition in a unit dosage form for ease of administration.

[0030] The compounds of the present invention, their tautomers, pharmaceutically acceptable salts or esters, and pharmaceutical compositions can be administered by any suitable method known in the art, including, but not limited to, oral, oral, sublingual, ocular, topical, parenteral, rectal, intrathecal, intracytoplasmic, inguinal, intravesical, topical (e.g., powders, ointments or drops), or nasal routes. However, for many therapeutic uses, the preferred route of administration / mode is parenteral administration (e.g., intravenous, subcutaneous, intraperitoneal, intramuscular). The technician will appreciate that the route of administration and / or mode will vary depending on the intended purpose. In a preferred embodiment, the compounds of the present invention, their tautomers, pharmaceutically acceptable salts or esters, and pharmaceutical compositions are administered by intravenous infusion or injection.

[0031] The pharmaceutical composition of the present invention may include a "therapeutically effective amount" or a "prophylactically effective amount" of a compound of the present invention, a pharmaceutically acceptable salt or ester thereof, a stereoisomer thereof, or a solvate thereof. A "prophylactically effective amount" refers to an amount sufficient to prevent, prevent, or delay the occurrence of a disease. A "therapeutically effective amount" refers to an amount sufficient to cure or at least partially prevent the disease and its complications in a patient already suffering from the disease. The therapeutically effective amount of a compound of the present invention, its tautomer, pharmaceutically acceptable salt or ester thereof may vary depending on the severity of the disease to be treated, the overall state of the patient's own immune system, the patient's general condition such as age, weight and sex, the mode of administration of the drug, and other treatments administered simultaneously, etc.

[0032] In the present invention, the dosage regimen can be adjusted to obtain the best intended response (e.g., a therapeutic or preventive response). For example, the dosage can be administered in a single dose, multiple doses can be administered over a period of time, or the dosage can be reduced or increased in proportion to the urgency of the treatment situation.

[0033] The typical non-limiting range of the therapeutic or prophylactic effective amount of the compound of the present invention, its tautomer, pharmaceutically acceptable salt or ester is 0.02-100 mg / kg, such as 0.1-100 mg / kg, 0.1-50 mg / kg, or 1-50 mg / kg. It should be noted that the dosage may vary depending on the type and severity of the symptoms to be treated. In addition, it is understood by those skilled in the art that for any particular patient, the specific dosing regimen should be adjusted over time according to the patient's needs and the doctor's professional evaluation; the dosage range given here is only for illustrative purposes and does not limit the use or scope of the pharmaceutical composition of the present invention.

[0034] In the present invention, the subject may be a mammal, such as a human.

[0035] The term "glucose metabolism disorders" refers to abnormalities in the structure, function, or concentration of hormones or enzymes that regulate the metabolism of glucose, fructose, galactose, etc., or pathophysiological changes in tissues and organs, and blood sugar monitoring will show elevated blood sugar levels.

[0036] The term "lipid metabolism disorders" refers to abnormalities in the substance and quantity of lipids (lipids) and their metabolites in the blood and other tissues and organs.

[0037] The term "metabolic syndrome" refers to a pathological state in which the body's proteins, fats, carbohydrates and other substances undergo metabolic disorders. It is a group of complex metabolic disorder syndromes and a risk factor for diabetes, cardiovascular and cerebrovascular diseases.

[0038] The term "obesity" refers to a certain degree of excess weight and excessive fat layer, especially excessive accumulation of body fat and triglycerides, which leads to a sub-healthy state. Excessive accumulation of body fat due to excessive food intake or changes in body metabolism causes excessive weight gain and causes pathological and physiological changes or latent changes in the human body.

[0039] The term "insulin resistance" refers to the decrease in the efficiency of glucose uptake and utilization promoted by insulin due to various reasons. The body compensatory secretion of excessive insulin produces hyperinsulinemia to maintain blood sugar stability, which in turn causes insulin resistance. Its main manifestations are high fasting blood sugar values ​​and slow response of the body to blood sugar in glucose tolerance tests. Obesity is one of the main causes of insulin resistance, especially central obesity. Obesity is mainly related to long-term lack of exercise and excessive dietary energy intake. 80% of patients with type 2 diabetes are obese when diagnosed. In the late stage of insulin resistance, insulin sensitivity further decreases, which can lead to the occurrence of diabetes. In the late stage of insulin resistance, it can gradually turn into diabetes. Detecting the effect of a compound on insulin resistance can indirectly reflect the effect of the compound on diabetes.

[0040] The term "hyperlipidemia" refers to a systemic lipid metabolism disorder characterized by elevated levels of plasma cholesterol, triglycerides, and low-density lipoprotein and low levels of high-density lipoprotein.

[0041] The term "hypercholesterolemia" refers to a cardiovascular disease caused by excessive cholesterol intake or cholesterol metabolism disorders, which leads to abnormally high serum total cholesterol and low-density lipoprotein concentrations. Hypercholesterolemia can increase blood viscosity, increase the burden on the heart, and cause poor operation of the microcirculatory system. In addition, hypercholesterolemia can accumulate on the blood vessel walls, leading to vascular atherosclerosis and ultimately vascular sclerosis. After fat accumulation, hypercholesterolemia can form fatty liver and even induce cerebral infarction.

[0042] The term "non-alcoholic fatty liver disease" refers to a clinical pathological syndrome characterized by excessive deposition of fat in hepatocytes caused by factors other than alcohol and other clear liver-damaging factors, and is involved in the pathogenesis of diseases such as insulin resistance and atherosclerosis.

[0043] The term "cardiovascular diseases" refers to a class of diseases involving the heart or blood vessels, and unhealthy diet and obesity, smoking, drinking, etc. are important causes. Through the high-fat mouse animal model, it was found that the compounds described herein (especially J2D) can significantly reduce the body weight, body fat percentage, liver wet weight, and the content of total triglycerides in liver tissue and serum of the model mice, indicating that the compounds can have a therapeutic effect on cardiovascular diseases. In some embodiments, cardiovascular diseases include coronary heart disease, such as angina pectoris and myocardial infarction (commonly known as heart disease), as well as stroke, heart failure, hypertensive heart disease, rheumatic heart disease, cardiomyopathy, arrhythmia, congenital heart disease, valvular heart disease, aortic aneurysm, peripheral arterial disease, thromboembolic disease and venous thrombosis. In some embodiments, when the cardiovascular disease specifically refers to vascular disease, it includes coronary artery disease, peripheral arterial disease, cerebrovascular disease, renal artery stenosis, aortic aneurysm, Raynaud's disease, Buerger's disease, peripheral venous disease, atherosclerosis, stroke, venous thrombosis and coagulation disorder. In some embodiments, when the cardiovascular disease specifically refers to heart disease, it includes cardiomyopathy, hypertensive heart disease, heart failure, cor pulmonale, arrhythmia, inflammatory heart disease (e.g., endocarditis, inflammatory myocardial hypertrophy, myocarditis, eosinophilic myocarditis), valvular heart disease, congenital heart disease or rheumatic heart disease. In some embodiments, the cardiovascular disease includes hypertension, hyperlipidemia (e.g., hypercholesterolemia), coronary heart disease, myocardial infarction and atherosclerosis.

[0044] The term "tautomer" refers to a functional group isomer resulting from the rapid movement of an atom in two positions in a molecule, such as enol-keto tautomers. In the present application, the compound of formula I may produce the following tautomerism, and the use of all possible tautomers in preventing or treating diseases related to lipid and / or sugar metabolism disorders is within the scope of protection of the present invention.

[0045]

[0046] Pharmaceutically acceptable salts of the compounds of the present invention include acid addition salts and base addition salts thereof.

[0047] For a review of suitable salts, see Stahl and Wermuth, "Handbook of Pharmaceutical Salts: Properties, Selection, and Use" (Wiley-VCH, 2002), or SM Berge et al., "Pharmaceutical Salts", J. Pharm. Sci. 1977, 66, 1-19. Methods for preparing pharmaceutically acceptable salts of the compounds of the invention are known to those skilled in the art.

[0048] Suitable acid addition salts are formed from acids that form pharmaceutically acceptable salts. Suitable base addition salts are formed from bases that form pharmaceutically acceptable salts. In some embodiments, suitable inorganic acids are such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, phosphoric acid or nitric acid, etc.; organic acids are such as formic acid, acetic acid, acetoacetic acid, pyruvic acid, trifluoroacetic acid, propionic acid, butyric acid, hexanoic acid, heptanoic acid, undecanoic acid, lauric acid, benzoic acid, salicylic acid, 2-(4-hydroxybenzoyl)-benzoic acid, camphoric acid, cinnamic acid, cyclopentanepropionic acid, digluconic acid, 3-hydroxy-2-naphthoic acid, nicotinic acid, pamoic acid, pectinic acid, 3-phenylpropionic acid , picric acid, pivalic acid, 2-hydroxyethanesulfonic acid, itaconic acid, aminosulfonic acid, trifluoromethanesulfonic acid, dodecylsulfuric acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, methanesulfonic acid, 2-naphthalenesulfonic acid, naphthalenedisulfonic acid, camphorsulfonic acid, citric acid, tartaric acid, stearic acid, lactic acid, oxalic acid, malonic acid, succinic acid, malic acid, adipic acid, alginic acid, maleic acid, fumaric acid, D-gluconic acid, mandelic acid, ascorbic acid, glucoheptonic acid, glycerophosphoric acid, aspartic acid, sulfosalicylic acid, etc. For example, HCl (or hydrochloric acid), HBr (or hydrobromic acid solution), methanesulfonic acid, sulfuric acid, tartaric acid or fumaric acid can be used to form a pharmaceutically acceptable salt with the compound shown in Formula I.

[0049] The term "pharmaceutically acceptable ester" means an ester derived from a compound of the invention, including physiologically hydrolyzable esters (which may be hydrolyzed under physiological conditions to release the compound of the invention in either free acid or alcohol form).

[0050] The term "excipient" refers to an excipient or vehicle used to administer the compound, including but not limited to diluents, disintegrants, precipitation inhibitors, surfactants, glidants, binders, lubricants, coating materials, etc. Examples of excipients include but are not limited to aluminum monostearate, aluminum stearate, carboxymethylcellulose, sodium carboxymethylcellulose, crospovidone, glyceryl isostearate, glyceryl monostearate, hydroxyethylcellulose, hydroxymethylcellulose, hydroxyoctadecanoate, hydroxypropylcellulose, hydroxypropylmethylcellulose, lactose, lactose monohydrate, magnesium stearate, mannitol, microcrystalline cellulose, etc.

[0051] The term "straight or branched C 2-10 "Alkyl" refers to a straight or branched chain alkyl group containing 2 to 10 (e.g., 2, 3, 4, 5, 6, 7, 8, 9 or 10) carbon atoms, such as C 2-8 Alkyl, C 4-8 Alkyl or C 4-6 Specific examples include, but are not limited to, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl or decyl.

[0052] The term "straight or branched C 2-10 "Alkenyl" refers to a straight or branched chain alkenyl group containing 2 to 10 (e.g., 2, 3, 4, 5, 6, 7, 8, 9 or 10) carbon atoms and at least one carbon-carbon double bond, such as C 2-8 Alkenyl, C 4-8 Alkenyl or C 4-6 Specific examples include, but are not limited to, vinyl, allyl, propenyl, butenyl, 2-butenyl, 3-butenyl, 1,3-dibutenyl, pentenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, 1,3-dipentenyl, 1,4-dipentenyl, 2,4-dipentenyl, hexenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl, 5-hexenyl, 1,3-dihexenyl, 1,4-dihexenyl, 1,5-dihexenyl, 2,4-dihexenyl, 2,5-dihexenyl, 3,5-dihexenyl, 1,3,5-trihexenyl, heptenyl, octenyl, nonenyl, or decenyl.

[0053] The term "straight or branched C 2-10 "Alkynyl" refers to a straight or branched chain alkynyl group containing 2 to 10 (e.g., 2, 3, 4, 5, 6, 7, 8, 9 or 10) carbon atoms and at least one carbon-carbon triple bond, such as C 2-8 Alkynyl, C 4-8 Alkynyl or C 4-6 Specific examples include, but are not limited to, ethynyl, propargyl, propynyl, butynyl, 2-butynyl, 3-butynyl, pentynyl, 2-pentynyl, 3-pentynyl, 4-pentynyl, hexynyl, 2-hexynyl, 3-hexynyl, 4-hexynyl, 5-hexynyl, heptynyl, octynyl, nonynyl, or decynyl.

[0054] Advantageous Effects of the Invention

[0055] The present inventors have discovered for the first time that polyketide compounds (especially J2D) can be used to prevent and treat diseases related to metabolic disorders (such as sugar and / or lipid metabolism disorders), which provides a new option for the treatment of the diseases and has important clinical application value. BRIEF DESCRIPTION OF THE DRAWINGS

[0056] Figure 1 Display the H NMR spectrum of compound J2D, test conditions:1 H-NMR (600 MHz, acetone-d6).

[0057] Figure 2 Display the J2D C NMR spectrum of compound, test conditions: 13 C NMR (150 MHz, acetone-d6)

[0058] Figure 3 Shows the effect of compound J2D on the glucose tolerance curve of the hyperlipidemia mouse model; among them: the model group is compared with the control group # P < 0.05, ## P < 0.01, ### P<0.001. Compared with the model group, * P < 0.05, ** P < 0.01, *** P<0.001, N=6.

[0059] Figure 4 Shown is the effect of compound J2D on fat vacuoles in liver slices of a high-fat mouse model.

[0060] Figure 5 Shows the liver sections of the control group, model group and drug-treated group in the high-fat fed mice experiment. DETAILED DESCRIPTION

[0061] The embodiments of the present invention will be described in detail below in conjunction with the examples, but those skilled in the art will appreciate that the following examples are only used to illustrate the present invention and should not be considered to limit the scope of the present invention. If no specific conditions are specified in the examples, they are carried out according to normal conditions or the conditions recommended by the manufacturer. If the manufacturer is not specified for the reagents or instruments used, they are all conventional products that can be obtained commercially.

[0062] Example 1 Preparation of Compounds

[0063] A person skilled in the art can prepare the compounds described in the present application by referring to the following methods, or obtain the compounds described in the present application by referring to the methods in the prior art, for example, Liu W, Gu Q, Zhu W, Cui C, Fan G. Dihydrotrichodimerol and tetrahydrotrichodimerol, two new bisorbicillinoids, from a marine-derived Penicillium terrestre. J Antibiot (Tokyo). 2005;58(10):621-4.

[0064] 1.1 Materials and Instruments

[0065] (1) Strain

[0066] The strain MMF4 (Acremonium citrinum) was isolated from sea mud (Aojiang Haikou Beach, Wenzhou).

[0067] (2) Experimental instruments:

[0068] Instrument Name Manufacturer DZF-6050 Vacuum Drying Oven Shanghai Jinghong Experimental Equipment Co., Ltd. <![CDATA[Bruker ASCEND TM 600 MHz NMR]]> Bruker, Switzerland Diaphragm vacuum pump Bruker, Switzerland Medium and low pressure chromatography system Bruker, Switzerland Mold incubator Shaoguang Keli Experimental Instrument Co., Ltd. Clean bench Suzhou Antai Air Technology Co., Ltd. Media Multi-function Induction Cooker Guangdong Midea Household Electrical Appliances Manufacturing Co., Ltd. LC-20A Analytical High Performance Liquid Chromatograph Shima Seiki N-1100 EYELA Rotary Evaporator Shanghai Ailang Instrument Co., Ltd. CA-1111 EYELA cooling water circulation device Shanghai Ailang Instrument Co., Ltd. SHB-III circulating water multi-purpose vacuum pump Zhengzhou Great Wall Science and Technology Trading Co., Ltd. Toledo XS205 DualRange Analytical Balance METTLER-TOLEDO, Switzerland PL602-L electronic balance METTLER-TOLEDO, Switzerland Milli-QB Ultrapure Water System Millipore Corporation

[0069] (3) Main reagents:

[0070] Reagent name Manufacturer Silica gel (200-300 mesh, thin layer silica gel) Qingdao Ocean Chemical Co., Ltd. TLC Plates Qingdao Ocean Chemical Co., Ltd. ODS (octadecylsilane bonded silica) Pharmacia Methanol Sinopharm Chemical Reagent Co., Ltd. Chloroform Sinopharm Chemical Reagent Co., Ltd. Deuterated reagents Cambridge CIL

[0071] (4) Culture medium

[0072] Rice medium: 100 g rice, 150 mL artificial seawater;

[0073] 1.2 Preparation of fermentation products

[0074] (1) Prepare 4800 g of rice culture medium;

[0075] (2) Cultivate in a 28°C mold incubator in the dark for 36 days.

[0076] 1.3. Preparation of crude strain extracts

[0077] (1) Add the extraction solvent (ethyl acetate:methanol = 4:1) to the fermentation product of the strain, stir, soak for 12 h, then filter, collect the filtrate, extract three times, and combine the filtrate to obtain the product;

[0078] (2) The filtrate is concentrated under reduced pressure to obtain a crude strain extract.

[0079] 1.4 Preparation of strain extracts

[0080] (1) extracting the crude strain extract with water:ethyl acetate (1:1), and concentrating the obtained ethyl acetate phase under reduced pressure to obtain the strain ethyl acetate extract;

[0081] (2) The ethyl acetate extract was then extracted with petroleum ether:methanol (1:1) to remove the oil components, and the resulting methanol phase was concentrated under reduced pressure to obtain the strain extract.

[0082] 1.5 ODS column chromatography, silica gel column chromatography, preparation of monomer compounds

[0083] (1) Dissolve the strain extract (10.0 g) in a small amount of methanol (5 mL) to obtain a methanol suspension of the crude strain extract;

[0084] (2) The suspension was evenly loaded on the top of the ODS column and gradient eluted with a methanol-water system (30:70, 50:50, 70:30, 100:0). Each gradient elution volume was 2000 mL. 19 fractions (Fr. 1-19) were obtained by TLC analysis.

[0085] (3) Fraction Fr. 15 was subjected to normal phase silica gel column chromatography with a mobile phase of petroleum ether-ethyl acetate system (1:0, 20:1, 17:1, 10:1, 5:1, 1:1, 0:1), and 8 fractions (Fr. 15A-15H) were obtained by merging according to TLC analysis;

[0086] (4) The fraction Fr. 15F was separated by semi-preparative HPLC using a chromatographic column (YMC PackODS-A 250×10 mm), a mobile phase of 60% acetonitrile, a flow rate of 4 mL / min, and a detection wavelength of 208 nm. The target compound J2D (Dihydrotrichodimerol) was finally obtained as a yellow powder with a molecular formula of C 28 H 34 O8, molecular weight is 498.23.

[0087]

[0088] J2D

[0089] 1H-NMR (600 MHz, acetone-d6) δ: 3.04 (1H, s, H-1), 1.37 (3H, s, 2-CH3), 1.42 (3H, s, 4-CH3), 3.15 (1H, s, H-7), 1.35 (3H, s, 8-CH3), 1.40 (3H,s, 10-CH3), 2.42 (1H, dt, J = 14.8, 7.4 Hz, H-2'), 2.57 (1H, dt, J = 15.4,7.7 Hz, H-2'), 2.26 (1H, m, H-3'), 5.48 (1H, m, H-4'), 5.49 (1H, m, H-5'),1.63 (3H, br d, J = 4.8 Hz, H-6'), 6.48 (1H, d, J = 14.8 Hz, H-2”), 7.28 (1H,dd, J = 14.8, 11.0 Hz, H-3”), 6.41 (1H, m, H-4”), 6.25 (1H, m, H-5”), 1.87 (3H, dd, J = 6.8, 1.6 Hz, H-6”).

[0090] 13 C NMR (150 MHz, acetone-d6) δ: 58.7 (C-1), 79.7 (C-2), 105.2 (C-3), 58.9 (C-4), 193.9 (C-5), 104.8 (C-6), 58.3 (C-7), 79.6 (C-8), 105.2 (C-9),60.3 (C-10), 201.5 (C-11), 104.6 (C-12), 193.6 (C-1'), 35.2 (C-2'), 29.0 (C-3'), 130.5 (C-4'), 126.5 (C-5'), 18.0 (C-6'), 176.1 (C-1”), 120.3 (C-2”),143.4 (C-3”), 132.0 (C-4”), 140.3 (C-5”), 18.9 (C-6”), 21.9 (2-CH3), 20.1 (4-CH3), 21.9 (8-CH3), 19.6 (10-CH3).

[0091] Example 2 Activity detection of compounds

[0092] 2.1 Main instruments

[0093] The body composition analyzer was EchoMRI-100H, the autofocus ultrasonic crusher was Covaris M220, the fully automatic biochemical analyzer was BS-240vet, the paraffin embedding machine was LEICA EG 1160, the upright microscope was Leica DM4B, the analytical balance was OHAUS, and the human blood glucose meter was a Roche blood glucose meter.

[0094] 2.2 Main Materials

[0095] Blood glucose test strips were purchased from Roche; serum total cholesterol, total triglyceride, high-density lipoprotein, and low-density lipoprotein detection kits were purchased from Kubel Company; high-fat feed (TP23400) and low-fat and low-sugar control feed (TP23402) were purchased from Nantong Trophy Co., Ltd.

[0096] 2.3 Animals

[0097] C57BL mice were purchased from the Experimental Animal Center of Xiamen University in accordance with the ethical standards for experimental animals. After the mice were 3 months old, the control group was given a low-fat and low-sugar control diet, and the model group was given a high-fat diet. After 3 months of modeling, the test substance was given by gavage at 3 mg / kg / d, and the control and model groups were given the same volume of solvent (0.5% sodium carboxymethyl cellulose). Samples were collected 1 month later to detect indicators related to obesity, insulin resistance, hypercholesterolemia, and non-alcoholic fatty liver disease.

[0098] 2.4 Observation indicators

[0099] 2.4.1 Body weight and body fat percentage before and after administration

[0100] The weight of mice was measured and recorded using an ordinary balance. The body composition analyzer detected the total body fat of mice based on the principle of nuclear magnetic resonance. The total body fat of mice / body weight was the body fat percentage.

[0101] 2.4.2 Liver wet weight

[0102] The liver can be divided into 5 lobes. The median fissure of the liver, which starts from the middle of the gallbladder fossa and extends upward and backward to the left wall of the inferior vena cava, is used as the boundary to divide the liver into left and right halves. The left and right halves of the liver are divided into the left lateral lobe, left medial lobe, right anterior lobe, right posterior lobe and caudate lobe by the interlobar fissure. The five lobes of liver tissue are completely peeled off, weighed and recorded, which is the total wet weight of the liver.

[0103] 2.4.3 Total triglyceride content in liver tissue homogenate

[0104] The liver of each mouse was accurately weighed in equal weight, and the liver was placed in an automatic focused ultrasonic disruptor for tissue disruption. The liver was centrifuged at 3500 rpm / 10 min, and the supernatant was collected. The total triglyceride content in the tissue homogenate was detected using a total triglyceride detection kit.

[0105] 2.4.4 Serum total cholesterol, total triglycerides, high-density lipoprotein, and low-density lipoprotein levels

[0106] The eyes of mice were removed and blood was collected. o C for 30 min, centrifuged at 3500 rpm / 10 min, collected the supernatant, and used an automatic biochemical analyzer to detect the contents of total cholesterol, total triglycerides, high-density lipoprotein, and low-density lipoprotein.

[0107] 2.4.5 Glucose tolerance test to detect blood sugar changes

[0108] The feed was removed but the water was not. After the mice were starved for 6 hours, the fasting blood glucose was measured, and the time was recorded as 0 min. A 25% glucose solution was prepared and 10 μL / g was given by gavage. The blood glucose values ​​at 15 min, 30 min, 60 min, and 90 min were measured respectively.

[0109] 2.4.6 Detection of fat vacuoles in liver tissue sections

[0110] The center of the left anterior lobe of the liver tissue of each mouse was taken for tissue section preparation. After fixation, dehydration, wax immersion, embedding, paraffin sectioning, dewaxing, hematoxylin / eosin staining, dehydration, wax immersion, sealing, upright microscope examination, multifunctional true color pathological image analysis software was used to count and analyze the fat vacuoles in the liver tissue sections.

[0111] 2.4.7 Statistical analysis

[0112] All data are expressed as mean ± standard error. The t-test was used for significant comparison between the two groups, and the statistical software was SPSS 16.0. When p < 0.05, it was considered to be significantly different, indicated by "*"; when p < 0.01, it was indicated by "**". When p < 0.001, it was indicated by "***".

[0113] 2.5 Results

[0114] 2.5.1 Effects of the drug-treated groups on body weight, body fat percentage, and liver wet weight

[0115] Table 1

[0116] Body weight before administration (g) Body weight after administration (g) Body fat percentage before administration (%) Body fat percentage after administration (%) Liver wet weight (g) Control group 29.6±1.748 27.5±2.427 13.8±2.47 13.6±1.69 1.8±0.12 Model Group <![CDATA[49.8±1.586 ### ]]> <![CDATA[49.8±2.874 ### ]]> <![CDATA[41.9±2.02 ### ]]> <![CDATA[41.6±1.61 ### ]]> <![CDATA[2.2±0.29 ## ]]> Drug group 48.2±0.943 <![CDATA[43.6±3.127 ** ]]> 41.4±2.007 <![CDATA[37.5±2.891 * ]]> <![CDATA[1.6±0.201 *** ]]>

[0117] Among them: the model group compared with the control group # P < 0.05, ## P < 0.01, ### P<0.001. Compared with the model group, * P < 0.05, ** P < 0.01, *** P < 0.001, N = 6

[0118] As shown in Table 1, the model group was compared with the control group in terms of body weight before administration, body weight after administration, body fat percentage before administration, body fat percentage after administration, and wet liver weight (P < 0.001), indicating that the model was successful. After administration, compared with the model group, the body weight and body fat percentage of the drug administration group decreased (P < 0.05), and the wet liver weight was significantly reduced (P < 0.001).

[0119] 2.5.2 Effects of total triglycerides in liver tissue and total cholesterol, total triglycerides, high-density lipoprotein, and low-density lipoprotein in serum of mice in the drug-treated group

[0120] Table 2

[0121] Total triglycerides in liver tissue Serum total cholesterol Serum total triglycerides Serum high-density lipoprotein Serum low-density lipoprotein Control group 0.8±0.279 5.6±1.67 0.8±0.199 1.1±0.189 0.2±0.125 Model Group <![CDATA[1.3±0.353 ### ]]> <![CDATA[9.9±1.54 ### ]]> 0.9±0.156 1.2±0.147 <![CDATA[0.4±0.0537 ## ]]> Drug group 1.1±0.201 8.8±0.727 <![CDATA[0.5±0.113 *** ]]> 1.2±0.123 <![CDATA[0.3±0.065 * ]]>

[0122] Among them: the model group compared with the control group # P < 0.05, ## P < 0.01, ### P<0.001. Compared with the model group, * P < 0.05, ** P < 0.01, *** P < 0.001, N = 6

[0123] As shown in Table 2, compared with the control group, the model group had significantly increased total triglycerides in liver tissue (P < 0.001) and significantly increased low-density lipoprotein in serum (P < 0.001), indicating that the model was successful. After administration, compared with the model group, the total triglycerides in liver tissue of the drug group were significantly reduced (P < 0.05), the low-density lipoprotein in serum was significantly reduced, and the total triglycerides were significantly reduced (P < 0.001), suggesting that the reason for the weight loss effect of the drug group may be through reducing total triglycerides.

[0124] 2.5.3 Effects of drug administration groups on glucose tolerance curve

[0125] The results are as follows Figure 3 As shown in the figure, the area under the glucose tolerance curve of the model group increased significantly compared with the control group (P < 0.001), suggesting that the model had insulin resistance. After administration, the area under the tolerance curve of the drug administration group was significantly reduced compared with the model group (P < 0.01), suggesting that the drug administration group may have an effect on insulin resistance.

[0126] 2.5.4 Effects of drug administration on fat vacuoles in liver slices

[0127] The results are as follows Figure 4 As shown, the model group ( Figure 4 B) and the control group ( Figure 4A) In contrast, the fat vacuoles in the liver tissue sections of the model group increased significantly, showing the characteristics of non-alcoholic fatty liver. Figure 4 C) Fat vacuoles were significantly reduced, indicating that the test substance has the effect of treating fatty liver. Figure 5 AR.

[0128] 2.6 Conclusion

[0129] The present invention detected the activity indexes related to lipid and sugar metabolism disorder-related diseases (such as metabolic syndrome, obesity, diabetes, insulin resistance, liver disease or cardiovascular disease) in the high-fat-fed mouse model group and the drug-administered group. The results showed that the compound of the present invention has the activity of preventing and treating metabolic syndrome, obesity, diabetes, prediabetes, insulin resistance, liver disease or cardiovascular disease.

[0130] Although the specific embodiments of the present invention have been described in detail, it will be understood by those skilled in the art that various modifications and substitutions may be made to those details based on all the teachings disclosed, and these changes are within the scope of protection of the present invention. The full scope of the present invention is given by the attached claims and any equivalents thereof.

Claims

1. Use of the following compounds, their tautomers or pharmaceutically acceptable salts, or pharmaceutical compositions containing any of the above ingredients in the preparation of drugs for preventing or treating diseases related to lipid and / or glucose metabolism disorders: The diseases related to lipid and / or sugar metabolism disorder are selected from obesity and fatty liver.

2. The use according to claim 1, wherein the fatty liver is non-alcoholic fatty liver.

3. The use according to claim 1 or 2, wherein the pharmaceutical composition contains the compound, its tautomer or pharmaceutically acceptable salt, and one or more pharmaceutically acceptable excipients.

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