CN112592379A

CN112592379A - beta-D-glucose short-chain fatty acid ester compound and preparation method and application thereof

Info

Publication number: CN112592379A
Application number: CN202011514632.1A
Authority: CN
Inventors: 范辉; 李小超; 杨欣
Original assignee: Guangdong Pharmaceutical University
Current assignee: Ruikang Oriental Medical Research Beijing Co ltd
Priority date: 2020-12-18
Filing date: 2020-12-18
Publication date: 2021-04-02
Anticipated expiration: 2040-12-18
Also published as: CN112592379B

Abstract

The invention discloses a beta-D-glucose short-chain fatty acid ester compound and a preparation method and application thereof, belonging to the technical field of organic synthesis. The compound is a compound shown in a formula I, or a stereoisomer, a pharmaceutically acceptable salt, a solvate or a prodrug of the compound shown in the formula I. Wherein R is methyl, ethyl, propyl, propylene, isopropylene, butyl, butylene, isobutylene, pentylene, or isopentylene. The compound has potential prevention and treatment effects on diabetes, hyperlipidemia, atherosclerosis, Alzheimer disease, cardiovascular and cerebrovascular diseases, inflammation, tumor and depression.

Description

beta-D-glucose short-chain fatty acid ester compound and preparation method and application thereof

Technical Field

The invention belongs to the technical field of organic synthesis, and particularly relates to a beta-D-glucose short-chain fatty acid ester compound and a preparation method and application thereof.

Background

The glycolipid metabolic diseases comprise hyperlipidemia, diabetes, fatty liver, obesity, arteriosclerotic cardiovascular and cerebrovascular diseases and other series diseases, the incidence rate of which is high, and the glycolipid metabolic diseases become one of the worldwide problems. In China, the proportion of people with hypercholesterolemia is 3.08 hundred million, that of people with diabetes is 1.4 hundred million, and that of people with early stage diabetes among adults is 50.1 percent higher, and the Chinese cardiovascular disease report 2016 indicates that 1 person dies of cardiovascular disease every 5 seconds. Diabetes, hyperlipidemia and other metabolic diseases and cardiovascular diseases are becoming increasingly beautiful and affecting the health of people in the world (Guo, Xiaoxue, Ronglu, etc.. glycolipid metabolic diseases and precise medicine [ J ]. world science and technology-traditional Chinese medicine modernization, 2017,19(1): 50-54.).

The pathophysiological basis for type II diabetes is insulin resistance and insulin hyposecretion. Research shows that the occurrence of insulin resistance is related to endotoxemia, chronic inflammatory reaction, short-chain fatty acid, bile acid metabolism and the like, and the intestinal flora of diabetic patients has obvious imbalance (Chinese research collaboration on insulin secretion, Anya Li-Shuang-Wei. Chinese research on insulin secretion and insulin resistance characteristics of newly diagnosed type 2diabetes patients [ J ]. Chinese journal of endocrine metabolism, 2008,24(3): 256-260). The short-chain fatty acid comprises acetic acid, propionic acid and butyric acid, is a product of carbohydrate which is not digested and decomposed by a host in intestinal tract by the zymolysis of intestinal flora, is an important energy source of intestinal epithelial cells, and can maintain the barrier effect of the intestinal mucosa; meanwhile, the protein is an important signal molecule and can directly activate G protein coupled receptor 41(GPR41) and GPR 43. Short Chain Fatty Acids (SCFA) bind to GPR43 to inhibit inflammatory responses, induce glucagon-like peptide 1(GLP-1) secretion from L cells in the distal small intestine and colonic epithelium, participate in the regulation of blood glucose homeostasis, and have a hypoglycemic effect. Experiments prove that the insulin resistance induced by high-fat diet is more prominent in mice lacking GPR43, and after the combination activation of SCFA and GPR41, the combination activation of SCFA and GPR41 can induce L cells to express gastrointestinal peptide hormone casein Peptide (PYY), inhibit intestinal tract motility and improve insulin sensitivity. These studies suggest that intestinal flora can affect the metabolism of the host by short-chain fatty acids, which are metabolites thereof, and that supplementation of a diet with a suitable amount of short-chain fatty acids can improve blood glucose levels and enhance insulin sensitivity (the study on the relationship between juan, cuckoo red intestinal flora and type 2diabetes has advanced [ J ] modern medicine and health, 2018(3): 398-.

Short-chain fatty acids (SCFA) have other important physiological functions, can reduce intestinal inflammatory reactions, improve intestinal epithelial barrier function, and have certain therapeutic effects on colon tumors, non-alcoholic fatty liver diseases, and obesity (xyloid, liuyuhong, zhuyu, chuyu, lilising, xujingdong. research progress of short-chain fatty acids in disease treatment [ J ] world china journal of digestion, 2017,25(13):1179 1186.).

Acetate and butyrate are the major SCFAs. Acetate reduces the number of marginal zone b (mzb) cells, promoting proliferation of islet autoantigen-reactive T cells. Butyrate can increase the number and function of regulatory T (T reg) cells, and can reduce insulitis and diabetes in mice (Li W, Wong F S. diabetes short-chain fatty acids protective against type 1diabetes [ J ]. Nature Immunology,2017,18(5): 484-486.).

Short Chain Fatty Acids (SCFA) can enhance intestinal barrier function, prevent inflammatory diseases caused by bacterial invasion, and help to suppress autoimmune diseases and type 1diabetes (T1D). SCFAs may also enhance intestinal barrier function, prevent inflammatory diseases caused by bacterial invasion, and help to suppress autoimmune diseases and type 1diabetes (T1D) (Kim C h. microbial or short-chain fatty acids: white diabetes? [ J ]. Cellular & Molecular Immunology,2017,15 (2)).

The Butyrate and Inulin combined have the effects of reducing blood pressure and blood sugar and losing weight, and have potential treatment effect on Patients with diabetic hypertension and hyperlipidemia (Roshanravan N, Mahdavi R, Alizadeh E, et al. Effect of butyl and insulin Supplementation on Glycemic State, Lipid Profile and glucose-Like Peptide 1Level in peptides with Type 2Diabetes: A random Double-bind, plasma-Controlled Trial [ J ]. Hormine & Metabolic Research,2017,49(11):886 + 891.).

In summary, SCFAs play an irreplaceable role in maintaining homeostasis and normal physiological metabolism.

Disclosure of Invention

Based on the important role of short-chain fatty acid in maintaining body homeostasis and normal physiological metabolism, the invention aims to provide a beta-D-glucose short-chain fatty acid ester compound with SCFAs activity on one hand.

The beta-D-glucose short-chain fatty acid ester compound also comprises derivatives thereof, and the derivatives are stereoisomers, tautomers, pharmaceutically acceptable salts, solvates or prodrugs of the beta-D-glucose short-chain fatty acid ester compound.

The invention also provides a pharmaceutical composition comprising a pharmaceutically acceptable carrier and at least the beta-D-glucose short chain fatty acid ester compound or a stereoisomer, a tautomer, a pharmaceutically acceptable salt, a solvate or a prodrug thereof.

In another aspect, the present invention also provides a method for preparing the beta-D-glucose short chain fatty acid ester compound or a stereoisomer, a tautomer, a pharmaceutically acceptable salt, a solvate, or a prodrug thereof.

In still another aspect, the invention also provides the application of the beta-D-glucose short-chain fatty acid ester compound in preparing medicaments for treating various glycolipid metabolic diseases, inflammations, tumors or cancers.

First, the present invention provides a β -D-glucose short-chain fatty acid ester compound having SCFAs activity.

The compound is shown as a formula I, or a stereoisomer, a pharmaceutically acceptable salt, a solvate or a prodrug of the compound shown as the formula I;

wherein R is alkyl or hydrogen; preferably, R is C_1-20An alkyl group; more preferably, R is C_1-10An alkyl group; still more preferably, said C_1-10Alkyl is C_1-10Straight chain alkyl or C_1-10A branched alkyl group; further preferably, R is hydrogen, methyl, ethyl, propyl, propylene, isopropylene, butyl, butylene, isobutylene, pentyl, pentylene, or isopentylene; still further preferably, said R is methyl, ethyl, propyl, butyl, isobutylene, pentyl or isopentylene; preferably, even more preferably, said R is methyl, ethyl or propyl.

The pharmaceutically acceptable salt is sodium salt, potassium salt, calcium salt or zinc salt.

The compound is glucose formate, glucose acetate, glucose propionate, glucose butyrate, glucose isobutyrate, glucose valerate or glucose isovalerate;

preferably, the compound is glucose acetate, glucose propionate or glucose butyrate.

Secondly, the invention provides a preparation method of the beta-D-glucose short-chain fatty acid ester compound, and the preparation method comprises the following steps: the glycosyl trichloroacetimidate is prepared by taking glucose as a raw material, short chain fatty acid reacts with the glycosyl trichloroacetimidate, and then benzyl is removed to obtain the target compound.

The method specifically comprises the following steps:

(1) refluxing a certain amount of sodium acetate and acetic anhydride, slowly adding glucose, reacting at 90 ℃ for 4h, cooling to room temperature, ultrasonically adding ice water, performing suction filtration to obtain a white solid, and recrystallizing with absolute ethyl alcohol to obtain white solid Glu 1;

(2) dissolving Glu1 obtained in step (1) in anhydrous dichloromethane, adding EtSH and BF at 0 deg.C₃OEt₂After the reaction at room temperature is finished, adding saturated sodium bicarbonate to stop the reaction, extracting with dichloromethane, washing with saturated sodium chloride aqueous solution, and spin-drying an organic layer to obtain yellow oily Glu 2;

(3) dissolving Glu2 obtained in the step (2) in absolute methanol, adding 30% sodium methoxide methanol solution, after the reaction is finished, adding Amberlite IR 120 to adjust the pH value to be neutral, filtering, spin-drying the solvent, and vacuum-drying to obtain Glu 3;

(4) dissolving Glu3 obtained in the step (3) in anhydrous DMF, adding NaH at 0 ℃, adding benzyl bromide for reaction, and carrying out silica gel column chromatographic separation to obtain Glu 4;

(5) dissolving Glu4 obtained in the step (4) in acetone/water solution, adding N-bromosuccinimide (NBS), after the reaction is completed, adding triethylamine in ice bath, spin-drying acetone, adding an equivalent amount of saturated sodium bicarbonate solution dissolved in dichloromethane for extraction, and washing an organic layer with saturated sodium chloride water solution to obtain Glu 5;

(6) dissolving Glu5 obtained in the step (5) in anhydrous dichloromethane, adding trichloroacetonitrile and 1, 8-diazabicyclo [5.4.0] undec-7-ene (DBU), and separating by silica gel column chromatography after the reaction to obtain Glu 6;

(7) dissolving the Glu6 obtained in the step (6) in anhydrous dichloromethane, adding R-COOH at-20 ℃, adding saturated sodium bicarbonate after complete reaction, extracting with dichloromethane, washing an organic layer with saturated sodium chloride, spin-drying the organic layer, and recrystallizing with diethyl ether-n-hexane to obtain Glu 7;

(8) dissolving the Glu7 obtained in the step (7) in 95% ethanol, adding 10% Pd-C, hydrogenating for 12h under normal pressure, centrifuging to remove Pd-C, spin-drying the filtrate, and recrystallizing with methanol-ethyl acetate to obtain Glu8, namely the beta-D-glucose short-chain fatty acid ester compound.

R in R-COOH in the above step (7) is C_1-20An alkyl group; still more preferably, R is C_1-10Alkyl radical, said C_1-10Alkyl is C_1-10Straight chain alkyl or C_1-10A pendant alkyl group; further preferably, R is hydrogen, methyl, ethyl, propyl, propylene, isopropylene, butyl, butylene, isobutylene, pentylene, or isopentylene; still further preferably, said R is methyl, ethyl, propyl, butyl, isobutyl, pentyl or isopentylene; preferably, even more preferably, said R is methyl, ethyl or propyl.

The preparation process comprises the following steps:

furthermore, the invention also provides a composition which comprises the compound shown as the formula I, the stereoisomer thereof, the pharmaceutically acceptable salt thereof or the solvate thereof, and a pharmaceutically acceptable auxiliary agent, carrier or diluent.

In some preferred embodiments, the composition is in a dosage form selected from plain tablets, film-coated tablets, sugar-coated tablets, enteric-coated tablets, dispersible tablets, capsules, granules, oral solutions or oral suspensions.

The invention further provides the application of the beta-D-glucose short-chain fatty acid ester compound or the stereoisomer thereof, or the pharmaceutically acceptable salt thereof or the solvate thereof in preparing a medicament for treating glycolipid metabolic diseases, neurological diseases, inflammations or cancers.

Wherein the glycolipid metabolic disease is hyperlipidemia, diabetes, fatty liver, obesity, hypertension or cardiovascular and cerebrovascular diseases;

the neurological disease is Parkinson syndrome, Alzheimer disease and depression; the inflammation is intestinal inflammatory disease, pneumonia or skin infectious disease.

The cancer is gastric cancer, cervical adenocarcinoma, colon cancer, lung cancer, liver cancer, glioma, esophageal cancer, intestinal cancer, nasopharyngeal cancer, breast cancer, lymphoma, kidney cancer, pancreatic cancer, bladder cancer, ovarian cancer, uterine cancer, bone cancer, gallbladder cancer, lip cancer, melanoma, tongue cancer, laryngeal cancer, leukemia, prostate cancer, brain tumor, squamous cell cancer, skin cancer, hemangioma, lipoma, cervical cancer or thyroid cancer.

The term "alkyl" as used herein is intended to include both branched and straight chain saturated hydrocarbon radicals having the specified number of carbon atoms. E.g. "C_1-10Alkyl "(or alkylene) groups are intended to be C1, C2, C3, C4, C5, C6, C7, C8, C9 and C10 alkyl groups. In addition, for example "C_1-10Alkyl "denotes an alkyl group having 1 to 10 carbon atoms. Alkyl groups may be unsubstituted or substituted such that one or more of its hydrogen atoms are replaced with another chemical group. Examples of alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (e.g., n-propyl and isopropyl), butyl (e.g., n-butyl, isobutyl, t-butyl), pentyl (e.g., n-pentyl, isopentyl, neopentyl), and the like. Accordingly, "alkylene" has one less hydrogen atom than "alkyl".

The compounds of the invention are understood to include both the free form and salts thereof, unless otherwise indicated. The term "salt" means an acid and/or base salt formed from an inorganic and/or organic acid and a base. In addition, the term "salt" may include zwitterions (internal salts), such as when the compound of formula I contains a basic moiety, such as an amine or pyridine or imidazole ring, and an acidic moiety, such as a carboxylic acid. Pharmaceutically acceptable (i.e., non-toxic, physiologically acceptable) salts are preferred, such as acceptable metal and amine salts, wherein the cation does not contribute significantly to the toxicity or biological activity of the salt. However, other salts may be useful, such as separation or purification steps in the preparation process, and are therefore included within the scope of the present invention. Salts of the compounds of formula I may be formed, for example, by combining a compound of formula I with an amount of acid or base, for example, in equal amounts, in a vehicle, for example, in which the salt precipitates or in which it is present in an aqueous vehicle, and then lyophilizing.

Exemplary acid addition salts include acetate (e.g., formed with acetic acid or trihaloacetic acid, such as trifluoroacetic acid), adipate, alginate, ascorbate, aspartate, benzoate, benzenesulfonate, bisulfate, borate, butyrate, citrate, camphorate, camphorsulfonate, cyclopentylpropionate, digluconate, dodecylsulfate, ethanesulfonate, fumarate, glucoheptonate, glycerophosphate, hemisulfate, heptanoate, hexanoate, hydrochloride (formed with hydrochloric acid), hydrobromide (formed with hydrobromic acid), hydroiodide, 2-hydroxyethanesulfonate, lactate, maleate (formed with maleic acid), methanesulfonate (formed with methanesulfonic acid), 2-naphthalenesulfonate, nicotinate, nitrate, oxalate, pectate, persulfate, 3-phenylpropionate, salts of benzoic acid, salts of, Phosphates, picrates, pivalates, propionates, salicylates, succinates, sulfates (such as those formed with sulfuric acid), sulfonates (such as those mentioned herein), tartrates, thiocyanates, tosylates such as tosylate, undecanoate, and the like.

Exemplary basic salts include ammonium salts, alkali metal salts such as sodium, potassium, calcium, and zinc salts; alkaline earth metal salts such as calcium and magnesium salts; barium, zinc and aluminum salts; salts with organic bases (e.g., organic amines) such as trialkylamines, e.g., triethylamine, procaine, dibenzylamine, N-benzyl- β -phenylethylamine, 1-diphenylhydroxymethylamine, N' -dibenzylethylenediamine, dehydroabietylamine, N-ethylpiperidine, benzylamine, dicyclohexylamine, or similar pharmaceutically acceptable amines and salts with amino acids such as arginine, lysine, and the like. Basic nitrogen-containing groups can be quaternized with agents such as lower alkyl halides (e.g., methyl, ethyl, propyl, and butyl chlorides, bromides, and iodides), dialkyl sulfates (e.g., dimethyl, diethyl, dibutyl, and diamyl sulfates), long chain halides (e.g., decyl, dodecyl, tetradecyl, and octadecyl chlorides, bromides, and iodides), aralkyl halides (e.g., benzyl and phenethyl bromides), and others. Preferred salts include monohydrochloride, bisulfate, mesylate, phosphate or nitrate salts.

The phrase "pharmaceutically acceptable" refers to those compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without additional toxicity, irritation, allergic response, or other problem or complication commensurate with a reasonable benefit/risk ratio.

As used herein, "pharmaceutically acceptable salts" refers to derivatives of the disclosed compounds wherein the parent compound is modified with an acid or its basic salt. Examples of pharmaceutically acceptable salts include, but are not limited to, inorganic or organic acid salts of basic groups such as amines; and acid groups such as bases or organic salts of carboxylic acids. Pharmaceutically acceptable salts include the conventional non-toxic salts or parent compounds forming quaternary ammonium salts, e.g. from non-toxic inorganic or organic acids. For example, such conventional non-toxic salts include those derived from inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, sulfamic acid, phosphoric acid and nitric acid; and salts prepared from organic acids such as acetic, propionic, succinic, glycolic, stearic, lactic, malic, tartaric, citric, ascorbic, pamoic, maleic, hydroxymaleic, phenylacetic, glutamic, benzoic, salicylic, sulfanilic, 2-ethoxybenzoic, fumaric, toluenesulfonic, methanesulfonic, ethane disulfonic, oxalic, and isethionic acid and the like.

The pharmaceutically acceptable salts of the present invention may be synthesized from parent compounds containing either a basic or acidic moiety by conventional chemical methods. Generally, these salts can be prepared from the free acid or base forms of these compounds with a stoichiometric ratio of the appropriate base or acid in water or an organic solvent, or a mixture of the two; generally, nonaqueous vehicles such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are preferred.

All stereoisomers of the compounds of the invention are contemplated, both in admixture or in pure or substantially pure form. Stereoisomers may include compounds that are optical isomers substituted by one or more chiral atoms, as well as compounds that are optical isomers by restricting the rotation of one or more bonds (atropisomers). The definition of the compounds of the present invention includes all possible stereoisomers and mixtures thereof. It includes especially the racemic form and the isolated optical isomers which have particular activity. Resolution of racemic forms by physical means, for example fractional crystallisation, separation or crystallisation of stereoisomeric derivatives or separation by chiral column chromatography. The individual optical isomers such as salts with optically active acids are obtained from the racemic salts by conventional methods and then crystallized.

Prodrugs and solvates of the compounds of the present invention are also contemplated. The term "prodrug" denotes a compound that undergoes a chemical reaction, either metabolically or chemically, upon administration to a receptor, to yield a compound of formula I, and/or a salt and/or solvate thereof. Any compound that is converted in vivo to provide a biologically active agent (i.e., a compound of formula I) is a prodrug within the scope and spirit of the invention. For example, compounds containing a carboxyl group may form physiologically hydrolyzable esters as prodrugs, which upon hydrolysis in vivo yield the compounds of formula I themselves. These prodrugs are preferably administered orally, since hydrolysis under many conditions occurs substantially under the influence of digestive enzymes. Parenteral administration can be used, the ester itself being active, in those instances hydrolysis occurs in the blood. Examples of physiologically hydrolyzable esters of compounds of formula I include C_1-6Alkylbenzyl, 4-methoxybenzyl, indanyl, phthaloyl, methoxymethyl, C_1-6alkanoyloxy-C_1-6Alkyl radicals such as acetoxymethyl, pivaloyloxymethyl or propoxymethyl, C_1-6Alkoxy carbonyloxy-C_1-6Alkyl radicals, such as methoxycarbonyl-oxymethyl or ethoxycarbonyloxymethyl, glycinyloxymethyl, phenylglycinyloxymethyl, (5-methyl-2-oxo-1, 3-dioxol-4-yl) -methyl and other well-known physiologically hydrolyzable esters of use, for example in the field of penicillin and cepalogensporin. These esters can be prepared by conventional techniques known in the art. Various forms of prodrugs are well known in the art.

"pharmaceutically acceptable carrier" refers generally to a carrier generally accepted in the art for delivering a biologically active agent to an animal, particularly a mammal. The pharmaceutically acceptable carrier is formulated according to a number of factors well known to those of ordinary skill in the art. These include without limitation the type and nature of the active agent being formulated; a subject to which a composition comprising the agent is administered; the route of administration of the composition; and directional therapy instructions. Pharmaceutically acceptable carriers include aqueous and non-aqueous liquid vehicles, and a variety of solid and semi-solid dosage forms. These carriers include many different components and additives in addition to the active agent, such additional components being included in the formulation for a variety of reasons, such as stability of the active agent, binder, etc., as is well known to those of ordinary skill in the art.

The compounds of formula I of the present invention may be administered in any suitable manner for treating a condition, depending on the site-specific treatment or the amount of drug delivered. Topical administration is generally preferred for systemic treatment of skin-related diseases, cancerous or pre-cancerous conditions, but other modes of delivery are contemplated. For example, orally administered compounds, such as in the form of tablets, capsules, granules, powders, or liquid formulations including syrups; topically such as in solution, suspension, gel or ointment; sublingual administration; the cheek floor; parenteral administration, e.g., by subcutaneous, intravenous, intramuscular or intrasternal injection or infusion (e.g., sterile aqueous or nonaqueous solution or suspension); nasal such as by inhalation spray; topically such as in the form of a lotion or ointment; rectally, e.g., in suppository form; or liposomal. Dosage unit formulations containing non-toxic, pharmaceutically acceptable excipients or diluents may be administered. The compound may be administered in an immediate release or delayed release form. Immediate release or delayed release may be achieved with suitable pharmaceutical compositions, in the case of partial delayed release, using devices such as subcutaneous implants or osmotic pumps.

Exemplary compositions for oral administration include suspensions which may contain, for example, microcrystalline cellulose for delivery, alginic acid or sodium alginate as a suspending agent, methylcellulose as a viscosity enhancer, and sweetening or flavoring agents such as those known in the art; immediate release tablets may contain, for example, microcrystalline cellulose, dicalcium phosphate, starch, magnesium stearate and/or lactose and/or other excipients, binders, extenders, disintegrants, diluents and lubricants such as those known in the art. The compounds of the invention may also be delivered orally by sublingual and/or buccal administration, e.g. compression molding, compression orAnd (4) freeze-drying the tablets. Exemplary compositions may include fast dissolving diluents such as mannitol, lactose, sucrose, and/or cyclodextrins. Included in these formulations may also be high molecular weight excipients such as cellulose

Or polyethylene glycol (PEG); excipients which aid mucosal adhesion such as hydroxypropyl cellulose (HPC), hydroxypropylmethyl cellulose (HPMC), sodium carboxymethylcellulose (SCMC) and/or maleic anhydride copolymers (e.g. HPMC)

) (ii) a And release controlling agents such as polyacrylic acid copolymers (e.g. CARBOPOL)

). Lubricants, glidants, flavors, colorants and stabilizers may also be added to aid in preparation and use.

Exemplary compositions for administration by spraying or inhalation include solutions which may contain benzyl alcohol or other suitable preservatives, absorption promoters to enhance absorption and/or biological activity, and/or other soluble or dispersible agents such as those known in the art.

Exemplary compositions for parenteral administration include injection solutions or suspensions which may contain, for example, suitable non-toxic, parenterally acceptable diluents or solvents, such as mannitol, 1, 3-butanediol, water, geline's solution, isotonic sodium chloride solution, or other suitable dispersing or wetting and suspending agents, including synthetic mono-or diglycerides, and fatty acids, including oleic acid.

Exemplary compositions for rectal administration include suppositories which may contain, for example, suitable non-irritating excipients such as cocoa butter, synthetic glycerides or polyethylene glycols which are solid at ordinary temperatures but dissolve and/or dissolve in the gastrointestinal tract to release the drug.

A therapeutically effective amount of a compound of the present invention can be determined by one of ordinary skill in the art and includes exemplary dosages of from about 0.05 to 1000 mg/kg; 1-1000 mg/kg; 1-50 mg/kg; 5-250 mg/kg; 250-1000mg/kg, which can be administered in a single dose or in separate divided doses, e.g. from 1 to 4 times daily, in terms of the amount of active compound per kg of body weight per day. It will be understood that the specific dose level and frequency of dosage for a particular subject may be varied depending upon a variety of factors including the activity of the specific compound employed, the metabolic stability and length of action of that compound, the race, age, body weight, general health, sex and diet of the subject, mode and time of administration, rate of excretion, drug combination and the severity of the particular disease. Preferred recipients for use in therapy include animals, most preferably mammalian races such as humans and poultry animals such as dogs, cats, horses and the like.

Detailed Description

The invention is further illustrated by the following examples. It should be understood that the method described in the examples is only for illustrating the present invention and not for limiting the present invention, and that simple modifications of the preparation method of the present invention based on the concept of the present invention are within the scope of the claimed invention. All materials and solvents used in the examples were purchased from Sigma Biochemical and Organic Compounds for Research and Diagnostic Reagents.

Example 1 a method for preparing a beta-D-glucose short-chain fatty acid ester compound-glucose formate ester includes the steps of:

(7) dissolving the Glu6 obtained in the step (6) in anhydrous dichloromethane, adding formic acid at-20 ℃, adding saturated sodium bicarbonate after complete reaction, extracting with dichloromethane, washing an organic layer with saturated sodium chloride, spin-drying the organic layer, and recrystallizing with diethyl ether-n-hexane to obtain Glu 7;

The structural formula of the glucose formate is as follows:

the nuclear magnetic hydrogen spectrum data of the glucose formate compound are as follows:

1HNMR(D2O):δ8.1(s,1H),5.20(d,1H,J＝9.8Hz),4.11(d,1H,J＝1.2Hz),3.74(m,1H),3.69(dd,1H,J＝2.8Hz),3.62(dd,1H,J＝3.0Hz),3.44(dd,1H,J＝9.7Hz)3.23(m,1H)。

example 2 preparation of beta-D-glucose short-chain fatty acid ester Compound-glucose acetate

The difference from example 1 is that: (7) dissolving the Glu6 obtained in the step (6) in anhydrous dichloromethane, adding acetic acid at-20 ℃, adding saturated sodium bicarbonate after complete reaction, extracting with dichloromethane, washing an organic layer with saturated sodium chloride, spin-drying the organic layer, and recrystallizing with diethyl ether-n-hexane to obtain Glu 7; other operations and steps are the same as in example 1.

The structural formula of the glucose acetate is as follows:

the nuclear magnetic hydrogen spectrum data of the glucose acetate compound are as follows:

¹HNMR(D2O):δ5.53(d,1H,J1,2 8.0Hz,H-1),4.01(dd,1H,J4,5 1.0Hz,H-4),3.86(m,1H,J5,6a 6.0,J5,6b 6.1Hz,H-5),3.79(m,2H,H-6a,b),3.76(dd,1H,J3,43.0Hz,H-3)3.75(dd,1H,J2,3 9.9Hz,H-2),2.23(s,3H,CH3-Ac)。

example 3 preparation of beta-D-glucose short-chain fatty acid ester Compound-glucose propionate

The difference from example 1 is that: (7) dissolving the Glu6 obtained in the step (6) in anhydrous dichloromethane, adding propionic acid at-20 ℃, adding saturated sodium bicarbonate after the reaction is completed, extracting the dichloromethane, washing an organic layer with saturated sodium chloride, spin-drying the organic layer, and recrystallizing with diethyl ether-n-hexane to obtain Glu 7;

other operations and steps are the same as in example 1.

The structural formula of the glucose propionate is as follows:

the nuclear magnetic hydrogen spectrum data of the glucose propionate compound are as follows:

1HNMR(D2O):δ5.40(d,1H,J＝8.1Hz),4.12(d,1H,J＝1.5Hz),3.78-3.70(m,3H),3.67(dd,1H,J＝9.2Hz)3.61(dd,1H,J＝2.4Hz),2.53(m,2H),1.75(dt,2H,J＝14.6,7.5Hz),0.98(dd,3H,J＝9.6t,5.2Hz)。

example 4A method for preparing beta-D-glucose short-chain fatty acid ester Compound-Glucobutyrate

The difference from example 1 is that: (7) dissolving the Glu6 obtained in the step (6) in anhydrous dichloromethane, adding butyric acid at-20 ℃, adding saturated sodium bicarbonate after complete reaction, extracting with dichloromethane, washing an organic layer with saturated sodium chloride, spin-drying the organic layer, and recrystallizing with diethyl ether-n-hexane to obtain Glu 7;

other operations and steps are the same as in example 1.

The structural formula of the glucose butyrate is as follows:

the nuclear magnetic hydrogen spectrum data of the glucose butyrate compound are as follows:

EXAMPLE 5 preparation of beta-D-glucose short-chain fatty acid ester Compound-glucose isobutyrate

The difference from example 1 is that: (7) dissolving the Glu6 obtained in the step (6) in anhydrous dichloromethane, adding isobutyric acid at-20 ℃, adding saturated sodium bicarbonate after complete reaction, extracting with dichloromethane, washing an organic layer with saturated sodium chloride, spin-drying the organic layer, and recrystallizing with diethyl ether-n-hexane to obtain Glu 7;

other operations and steps are the same as in example 1.

The structural formula of the glucose isobutyrate is as follows:

the nuclear magnetic hydrogen spectrum data of the glucose isobutyrate compound are as follows:

1HNMR(D2O):5.64(d,1H,J＝8.7Hz),4.13(d,1H,J＝1.2Hz),3.75(m,1H),3.69(dd,1H,J＝2.4Hz),3.62(dd,1H,J＝1.3Hz),3.44(dd,1H,J＝9.2Hz),3.37(d,1H,J＝3.2Hz),3.21(m,1H),1.12(d,6H,J＝14.6Hz)。

EXAMPLE 6 preparation of beta-D-glucose short-chain fatty acid ester Compound-Gluconate valerate

The difference from example 1 is that: (7) dissolving the Glu6 obtained in the step (6) in anhydrous dichloromethane, adding valeric acid at-20 ℃, adding saturated sodium bicarbonate after complete reaction, extracting with dichloromethane, washing an organic layer with saturated sodium chloride, spin-drying the organic layer, and recrystallizing with diethyl ether-n-hexane to obtain Glu 7;

other operations and steps are the same as in example 1.

The glucose valerate has a structural formula as follows:

the nuclear magnetic hydrogen spectrum data of the glucose valerate compound are as follows:

1HNMR(D2O):δ5.74(d,1H,J＝7.8Hz),4.07(d,1H,J＝1.0Hz),3.85(m,1H),3.69(dd,1H,J＝2.4Hz),3.57(d,1H,J＝1.8Hz),3.44(dd,1H,J＝9.2Hz),3.30(dd,1H,J＝2.7Hz),2.38(t,2H,J＝12.8),1.75(m,2H),1.38(m,2H),0.93(dd,3H,J＝10.7,5.8Hz)。

example 7A method for preparing beta-D-glucose short-chain fatty acid ester Compound-glucose Isovalerate

The difference from example 1 is that: (7) dissolving the Glu6 obtained in the step (6) in anhydrous dichloromethane, adding isovaleric acid at-20 ℃, adding saturated sodium bicarbonate after complete reaction, extracting with dichloromethane, washing an organic layer with saturated sodium chloride, spin-drying the organic layer, and recrystallizing with diethyl ether-n-hexane to obtain Glu 7;

other operations and steps are the same as in example 1.

The structural formula of the glucose isovalerate is as follows:

the nuclear magnetic hydrogen spectrum data of the glucose isovalerate compound are as follows:

1HNMR(D2O):5.57(d,1H,J＝8.2Hz),3.94(d,1H,J＝1.6Hz),3.69(dd,1H,J＝2.4Hz),3.68(d,1H,J＝2.9Hz),3.64(dd,1H,J＝8.2Hz),3.44(dd,1H,J＝2.7Hz),3.13(d,1H,J＝12.7Hz),2.41(t,2H,J＝10.4),2.40(m,1H),0.94(d,6H,J＝4.8Hz)。

effect test:

1. the 7 beta-D-glucose short-chain fatty acid ester compounds can promote and improve the glycolipid metabolic disturbance of rats, improve the insulin sensitivity, promote the secretion of the intestinal hormone GLP-1 and prevent and treat diabetes.

Experimental methods

(1) Animal model: male SD rats 180 are bred adaptively for 1 week, 10 male SD rats are randomly selected as a normal group and fed with a common feed; the remaining 170 rats were fed a high fat diet (20% sucrose, 2.5% cholesterol, 10% lard, 0.2% sodium cholate, 67.3% basal diet). After feeding for 4 weeks, the animals in the high-fat-fed group were fasted for 12 hours, and injected intraperitoneally with Streptozotocin (STZ) at 30 mg/kg-1. Rats were fed with normal diet and injected with an equal volume of citric acid-sodium citrate buffer. After 3 days, the blood sugar after fasting for 16 hours is detected, and the fasting blood sugar of 16.7mmol/L is taken as the successful standard of the diabetes molding.

(2) Experiment grouping

150 rats meeting the requirements of the diabetes model are randomly grouped according to blood sugar and body weight, and a low dose group (0.1 g/kg) and a high dose group (1.0 g/kg) of 7 compounds of glucose formate (GOF), glucose acetate (GOA), glucose propionate (GOP), glucose butyrate (GOF), glucose isobutyrate (GOIB), glucose valerate (GOV) and glucose isovalerate (GOIV) are set, wherein 10 rats are selected from each group, 10 rats are selected from the diabetes model group, and 10 rats are selected from a normal group fed with normal feed.

(3) Measurement index

3.1 blood glucose concentration determination: blood is collected from the tail of a rat, fasting plasma glucose (FBG) of the animal is measured by a glucometer and test paper, and blood glucose is measured after 4 weeks of administration. The test is started 8 hours in the morning without water supply.

3.2 sugar tolerance test (OGTT): rats were fasted for 12h before the experiment, and were gavaged with 50% glucose at 2 g/kg. Before (0min) and 30, 60 and 120min after intragastric administration of glucose, blood glucose at each time point is respectively measured, an OGTT curve is drawn, and the area under the curve (AUC) is calculated.

3.3 intestinal hormone glucagon-like peptide-1 (GLP-1) assay: adopting a glucagon-like peptide 1(GLP-1) detection kit: the whole blood sample collected in the serum separation tube is left at room temperature for 2 or 4 hours overnight, and then centrifuged at 1000 Xg for 20 minutes, and the supernatant is collected.

(4) Measurement results

4.1 Effect on serum fasting plasma glucose (FBG) in diabetic rats

As shown in Table 1, the FBG levels of the animals in each group before modeling have no obvious difference (P is more than 0.05), and the test results show that the blood sugar level of the rats in the model group is obviously higher than that of the rats in the normal group (P is less than 0.05) when the drug intervention is not carried out, which indicates that the experimental diabetic rat model is successfully established in the research.

Given GOF, GOA, GOP, GOB, GOIB, GOV, GOIV low dose group and high dose group, the results show: the levels of FBG in rats in the GOP low-dose group and the high-dose group after 4 weeks are obviously reduced compared with the levels before administration, and the difference of the levels in the GOP low-dose group and the high-dose group compared with the model group has statistical significance (P is less than 0.05); the levels of FBG in rats after 4 weeks in the low-dose group and the high-dose group of the GOB are obviously reduced compared with the levels before administration, and the difference of the FBG levels in rats after administration of the GOB and the GOB in the high-dose group is statistically significant (P is less than 0.05); the FBG level of rats in the low-dose group and the high-dose group of the GOIV is obviously reduced after 4 weeks compared with that before administration, and compared with the model group, the difference has statistical significance (P is less than 0.05), and the results show that the fasting blood glucose curative effect of the rats with diabetes treated by GOB, GOP and GOIV is obvious, and a dose dependence relationship exists.

Table 1 comparison of FBGs in groups of rats (x ± S, n ═ 10)

Note: p <0.05 compared to normal group and P <0.05 compared to model group.

4.2 Effect on diabetic rat glucose tolerance (OGTT)

As shown in Table 2, after the same dose of glucose was taken orally, all rats showed a glucose absorption and metabolism process with a first rise in blood glucose and then a gradual decline in blood glucose. After modeling, the blood sugar of the diabetic rat OGTT at each time point is obviously higher than that of a normal rat (P is less than 0.01), which shows that the glucose tolerance of the rat can be obviously reduced by combining high-fat feed with small-dose STZ injection, and a T2DM model is successfully copied.

After 4 weeks of administration, the blood glucose was significantly higher in the OGTT test than in the normal group at each time point in the model group, indicating that this T2DM model was feasible. Compared with the model group rats, the blood sugar level of the high-low dose group of GOB shows a reduction trend, is obvious at 120min, the AUC area is obviously reduced, and the difference has statistical significance (P is less than 0.05). The effect of improving the OGTT of the GOB high-dose group is better than that of the GOB low-dose group, and the effect of improving the OGTT of other GOB high-dose groups and the effect of improving the OGTT of the GOIV high-dose group are statistically different (P is less than 0.05). Indicating that the GOB high and low dose group improves the OGTT of the STZ diabetic rats.

TABLE 2 comparison of OGTT in rats of various groups after administration (x + -S, n ═ 10)

Note: AUC area compared to normal group, # P < 0.01, compared to model group, # P < 0.05.

4.3 measurement of glucagon-like peptide-1 (GLP-1), an intestinal hormone of diabetic rats

Table 3 shows that the GLP-1 level in STZ hyperglycemic rats is obviously lower than that in a normal group, the content of GLP-1 can be increased in the group with low GOB and high GOB, the GLP-1 level can be increased in the other group with high GOA, low GOP, high GOV and low GOIV, statistical differences exist, and the results indicate that part of beta-D-glucose short-chain fatty acid ester compounds improve beneficial intestinal hormones.

Table 3 comparison of serum GLP-1 content in rats of each group after administration (x ± S, n ═ 10)

Note: p < 0.01 compared to normal group and P <0.05 compared to model group.

2. Effect of 7 beta-D-glucose short-chain fatty acid ester compounds on reducing in-vitro hepatocyte steatosis 1 experiment method

1.1 establishment of FFA-induced HepG2 cell steatosis model

When HepG2 cells were plated on the bottom of the dish, 2X 10 cells per well⁵The density of individual cells was seeded in 6-well plates at 37 ℃ with 5% CO₂After 48h incubation under conditions, the experiments were grouped, i.e.: normal control group, model group, GOF, GOA, GOP, GOB, GOIB, GOV, GOIV low concentration group 10 μmol/mL and high concentration group 100 μmol/mL. Normal control group: culturing with DMEM medium containing 10% fetal calf serum; model group: adding culture medium containing FFA for culture; the administration group is added with FFA-containing culture medium and compounds with high and low concentrations. The culture was carried out for 24h under each experimental condition, and each group was provided with 6 multiple wells.

1.2 determination of Triglyceride (TG)/protein content

(l) Removing the residual culture solution in the wells, washing for 1-2 times by PBS, adding a trypsin solution to digest cells, preparing cell suspensions, averagely dividing the cell suspensions in the wells into two parts (cells I and cells II), and respectively moving the cell suspensions into 1.5mL EP tubes;

(2) washing the cells twice with PBS at 4 ℃, centrifuging for 5min at 1000rpm at 4 ℃, and removing the supernatant;

(3) the residual liquid in the EP tube is completely sucked on the absorbent paper, the cells are added into the tube firstly, 100 mu L of isopropanol is added into the tube, and the mixture is shaken and evenly placed in an ice box; adding 100 mu LPBS into the cell ② tube, shaking uniformly and placing in an ice box;

(4) the cells are cracked by an ultrasonic cell crusher at the temperature of 4 ℃, centrifuged at 13500rpm for 10min at the temperature of 4 ℃, the supernatant is collected, the TG content in the supernatant of the cells is measured by a triglyceride kit (strictly determined according to the operation of the specification), and the protein content of the cells is measured by a BCA protein quantitative kit (strictly determined according to the operation of the specification), and the protein content is converted into the ratio of TG/protein content.

2. Results of the experiment

As shown in Table 4, high and low concentrations of GOF, GOA, GOP, GOB, GOIB, GOV, and GOIV (10. mu. mol/mL, 100. mu. mol/mL) were applied to HepG2 cells for 24 hours, and the intracellular triglyceride content was measured using a triglyceride kit. The results show that the intracellular triglyceride content is significantly higher in the model group compared to the blank group; compared with the model group, the GOB high-low concentration group, the GOP high-low concentration group, the GOV high-low concentration group, the GOIV high-low concentration group, the GOF high-concentration group and the GOA high-concentration group can obviously reduce the content of triglyceride in cells, and the difference has statistical significance (p is less than 0.05); the reduction effect of the GOB high-dose group is larger than that of the GOB low-dose group, and the reduction effect of the 7 beta-D-glucose short-chain fatty acid ester compounds is suggested to have the effect of reducing in-vitro liver cell fatty degeneration, wherein the GOB has the strongest inhibition capacity and can inhibit FFA (fat deposition in cells) in a dose-dependent manner.

TABLE 4 Effect of high and low concentrations of GOF, GOA, GOP, GOB, GOIB, GOV, GOIV on triglyceride accumulation in liver cells (GOA), (GOB), (GOIB), (GOV), (GOIV)

n＝6)

Note: p < 0.01 in comparison with normal group, tangle-solidup <0.05 in comparison with model group, tangle-solidup < 0.01.

3. 7 beta-D-glucose short-chain fatty acid ester compounds with effect of reducing in vitro inflammation

1 method of experiment

LPS induces an in vitro model of inflammation and intervention of β -D-glucose short chain fatty acid ester compounds: the mouse macrophage J774 cell line is stimulated by 100ng/mL LPS to induce inflammatory response for 1h, and intervention is carried out by utilizing high and low concentrations of GOF, GOA, GOP, GOB, GOIB, GOV and GOIV (10 mu mol/mL and 100 mu mol/mL). After the experiment is finished, collecting cells and determining the expression levels of inflammatory factors interleukin 1 beta (IL-1 beta), tumor necrosis factor alpha (TGF-alpha) and monocyte chemotactic factor 1(MCP-1) by an RT-q PCR method.

2. Results of the experiment

As shown in Table 5, the expression levels of inflammatory factors IL-1. beta., TNF. alpha. and MCP-1 were measured by RT-q PCR in which high and low concentrations of GOF, GOA, GOP, GOB, GOIB, GOV and GOIV (10. mu. mol/mL and 100. mu. mol/mL) were applied to an in vitro inflammation model induced by LPS. The results show that the expression levels of IL-1 beta, TNF alpha and MCP-1 are obviously up-regulated (P is less than 0.05) compared with the normal group when the J774 cells are stimulated by LPS for 1 h; compared with the model group, the GOA high-low concentration group, the GOP high-low concentration group, the GOB high-low concentration group, the GOIV high-low concentration group, the GOF high-concentration group and the GOA high-concentration group can obviously reduce the expression of IL-1 beta, TNF alpha and MCP-1, and the difference has statistical significance (p is less than 0.05); among them, GOA and GOB have the strongest inhibitory ability, suggesting that 7 beta-D-glucose short-chain fatty acid ester compounds have anti-inflammatory effect.

TABLE 5 Effect of high and Low concentrations of GOF, GOA, GOP, GOB, GOIB, GOV, GOIV on LPS-induced in vitro inflammation ((GOA), (GOB), and (GOIV))

n＝6)

In conclusion, 7 compounds of glucose formate (GOF), glucose acetate (GOA), glucose propionate (GOP), glucose butyrate (GOF), glucose isobutyrate (GOIB), glucose valerate (GOV) and glucose isovalerate (GOIV) prepared by the embodiment of the invention have the effects of promoting and improving rat glycolipid metabolic disorder, improving insulin sensitivity, promoting secretion of intestinal hormone GLP-1 and preventing and treating diabetes; and has the effect of reducing the steatosis in liver cells in vitro; has effect in inhibiting inflammation.

The present invention has been further described with reference to specific embodiments, which are only exemplary and do not limit the scope of the present invention. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention, and that such changes and modifications may be made without departing from the spirit and scope of the invention.

Claims

1. A compound, which is a compound shown in formula I, or a stereoisomer, a pharmaceutically acceptable salt, a solvate or a prodrug thereof of the compound shown in formula I;

wherein R is C_1-20Alkyl or hydrogen.

2. The compound of claim 1, wherein: r is C_1-10Alkyl or hydrogen; said C_1-10Alkyl is C_1-10Straight chain alkyl or C_1-10A branched alkyl group.

3. The compound of claim 2, wherein: and R is H, methyl, ethyl, propyl, propylene, isopropylene, butyl, butylene, isobutylene, amyl, amylene or isoamylene.

4. A compound according to claim 3, characterized in that: r is methyl, ethyl, propyl, butyl, isobutyl, amyl or isoamylene; preferably, said R is methyl, ethyl or propyl.

5. A process for preparing a compound according to any one of claims 1 to 4, characterized in that: the glycosyl trichloroacetimidate is prepared by taking glucose as a raw material, short chain fatty acid reacts with the glycosyl trichloroacetimidate, and then benzyl is removed to obtain the target compound.

6. The method of claim 5, wherein the reaction is as follows:

7. the method according to claim 6, characterized by the following specific steps:

8. A composition characterized by: the composition comprising a compound, a stereoisomer of the compound, a pharmaceutically acceptable salt of the compound, or a solvate of the compound of any one of claims 1-5, and a pharmaceutically acceptable adjuvant, carrier, or diluent.

9. Use of a compound of any one of claims 1-5, a stereoisomer of said compound, a pharmaceutically acceptable salt of said compound or a solvate of said compound, or a composition of claim 8, in the manufacture of a medicament for the treatment of a glycolipid metabolic disease, a neurological disease, inflammation or cancer.

10. Use according to claim 9, characterized in that: the glycolipid metabolic disease is hyperlipidemia, diabetes, fatty liver, obesity, hypertension or cardiovascular and cerebrovascular diseases;

the neurological disease is Parkinson syndrome, Alzheimer disease and depression; the inflammation is intestinal inflammatory disease, pneumonia or skin infectious disease;