CN1668311A

CN1668311A - Method and composition for treating diabetes

Info

Publication number: CN1668311A
Application number: CNA028296354A
Authority: CN
Inventors: 陈小茁; 李运生; 李京; 刘芳
Original assignee: Ohio State University
Current assignee: Ohio State University
Priority date: 2002-07-24
Filing date: 2002-07-24
Publication date: 2005-09-14
Also published as: JP2005538987A; WO2004009094A1; EP1545554A1; CA2496912A1; EP1545554A4; AU2002322623A1

Abstract

Methods for modulating diabetes, impaired glucose tolerance, gestational diabetes and glucose resistance in a mammal, particularly a human. In one embodiment the method comprises administering a gallotannin composition to a mammal in need of the same. The gallotannin composition comprises one or more select hydrolysable gallotannins. In another embodiment the method comprises administering a gallotannin variant composition comprising one ore more select gallotannin variant compounds to the subject. Methods of preventing or treating weight gain in a subject. The method comprises administering the gallotannin composition of the present invention, the gallotannin variant composition of the present invention, or a combination of the gallotannin composition of the present invention and the gallotannin variant composition of the present invention to the subject. The present invention also relates a gallotannin variant compound or a salt thereof, and a pharmaceutical composition comprising such compound or the salt thereof.

Description

The method and composition of treatment diabetes

Invention field

The present invention relates to regulate the method and composition of diabetes and other other disease relevant in mammal with the glucose of abnormality and/or insulin level.This method is used the compositions of not induced lipolysis generation or hypoglycemia.

Background technology

Diabetes are commonly referred to diabetes (diabetes), are meant a kind of lysis that is caused by multiple factor, it is characterized in that the plasma glucose of improving the standard being referred to as hyperglycemia.Referring to as LeRoith, D. etc. (volume), DIABETES MELLITUS (Lippincott-Raven Publishers, Philadelphia, Pa.U.S.A., 1996) and whole documents of wherein quoting.According to ADA, estimate diabetes affects about global human 6%.Owing to suffer from blood capillary and trunk disease, comprise that the danger of nephropathy, neuropathy, retinopathy, hypertension, cerebrovascular disease and coronary heart disease improves, unsteered hyperglycemia is with raising relevant with too early mortality rate.Therefore, the control glucose homeostasis is the important method of treatment diabetes.

The diabetes that two kinds of principal modes are arranged, type 1 diabetes (being called insulin-dependent diabetes or IDDM in the past) and type 2 diabetes mellitus (being called noninsulindependent diabetes or NIDDM in the past).Type 1 diabetes is to lack insulin (regulating the hormone of glucose utilization) fully to cause.It is feature that insulin deficit damages with the pancreatic in pancreas usually, and insulin lacks fully.Type 2 diabetes mellitus be a kind of be the disease of feature with the insulin resistance, be accompanied by relative, rather than absolute insulin deficit.Type 2 diabetes mellitus can comprise from outstanding insulin resistance follows relative insulin deficit, follows some insulin resistances to outstanding insulin deficit.Insulin resistance is insulin have been reduced in wide concentration range bring into play its bioactive ability.In the insulin resistance individuality, the insulin of the amount that the health secretion is unusual high is to compensate this defective.When existing islets of langerhans in shortage usually to compensate insulin resistance and fully controlling glucose, the glucose tolerance status of weakening has taken place.In a large amount of individualities, insulin secretion further descends, and plasma glucose levels rises, and causes diabetes clinically.

Most of type 2 diabetes mellitus patients are by by the blood sugar lowering that stimulates β cell uelralante to play a role, or by can strengthening the medicine of patient to the structure sensitive properties of insulin, or islets of langerhans is usually treated.The sulphur urea is an example, can stimulate β cell uelralante.In strengthening the medicine of patient to the structure sensitive properties of insulin, metformin is representative example.Even the sulphur urea is widely used in the treatment type 2 diabetes mellitus, in most of the cases its treatment is satisfied inadequately.In a large amount of type 2 diabetes mellitus patients, sulphur urea deficiency is so that blood sugar level normalization, and therefore, the patient has the high risk of suffering from diabetic complication.Equally, many patients little by little lose the reaction to the treatment of sulphur urea, thereby, little by little be forced to carry out insulinize.The patient treats insulinize exhausting owing to pancreatic beta cell among the type 2 diabetes mellitus patients usually from oral hypoglycemic.

Insulin stimulating is by skeletal muscle and fatty tissue ingestion of glucose, mainly inserts to cell surface (Saltiel, A.R.﹠amp by glucose transporter 4 (GLUT4) storage site in the cell; Kahn, C.R. (2001) Nature 414:799-806; Saltiel, A.﹠amp; Pessin, J.E. (2002) Trends in Cell Biol.12:65-71; White, M.F. (1998) Mol.Cell.Biochem.182:3-11).In order to respond insulin, the GLUT4 component that is present in the cell inner membrance redistributes on the plasma membrane, causes the raising of GLUT4 on cell surface and the glucose uptake of cell to strengthen.The GLUT4 transposition is mainly by Insulin receptor INSR (IR) mediation.

Except glucose transport, insulin and lipogenesis are closely related, this process comprise along with fat in the cell inner accumulated, the propagation and the preceding adipose cell of preceding adipose cell (preceding-adipose cell) are divided into adipose cell (adipose cell).To studies show that of adipose cell lines 3T3-L1, the effect of insulin in lipogenesis mainly is mitosis (43).Before differentiation, the 3T3-L1 cell is the preceding adipose cell of fibroblast-sample, comprises more IGF-1 receptor than IR.External, the lipogenesis of preceding adipose cell can be generally used for inducing cocktail (cocktail) MDI of differentiation to cause by a kind of, and it is by a kind of reagent methyl-isobutyl xanthine (MIX) that improves cAmp; A kind of glucocorticoid dexamethasone (DEX); And with preceding adipose cell on the insulin (or IGF-1) of IGF-1 receptor response form (Tong, Q., Hotamisligil, G.S. (2001) Rev.in Endoc.﹠amp; Metabolic Disorders.2:349-355; Rosen, E.D. etc., (2000) Genes Dev.14:1293-1307).When treating with MDI, the preceding adipose cell that converges reenters cell cycle, and experiences about 2 mitosiss of taking turns (Modan-Moses, D. etc. (1998) Biochem.J.333:825-831; Tong, Q.Hotamisligil, G.S. (2001) Rev.in Endoc.﹠amp; Metabolic Disorders.2:349-355; Rosen, E.D. waits (2000) Genes Dev.14:1293-1307), this process is often referred to clone's expansion.After clone's expansion, preceding adipose cell withdraws from cell cycle, and comprises C/EBP-α, β, δ and PPAR-γ adipose cell gene by expression, begins to be divided into adipose cell.

Because having, its lipogenesis effect, insulin in the type 2 diabetes mellitus patient, promote fat harmful effect (referring to Moller, D.E. (2001) Nature 414:821-827).Unfortunately, other is used to also have the lipogenesis activity at the anti--diabetes medicament of type 2 diabetes mellitus patient moderate stimulation glucose transport at present.Thereby, though present Drug therapy can make blood glucose reduce, often promote fat.So, be starved of a kind of new anti--diabetes medicament of exploitation, can correct hyperglycemia and do not cause the lipogenesis side effect of following generation.Can in diabetics, induce glucose uptake and not cause that hypoglycemic chemical compound also needs.

Summary of the invention

The invention provides mammal, philtrum is especially regulated the method for glucose tolerance, gestational diabetes and the glucose resistance of diabetes, weakening.In an embodiment of this method, comprise to a kind of compositions of the administration of needs, be called " gallotannin (gallotannin) compositions " here.The gallotannin compositions is fully pure, comprises one or more hydrolyzable gallotannins, is selected from: 1,2,3, and 4-four-O-galloyl-alpha-D-glucose, 1,2,3,6-four-O-galloyl-alpha-D-glucose, 1,3,4,6-four-O-galloyl-alpha-D-glucose, 1,2,3,4,6-five-O-galloyl-alpha-D-glucose, 1,2,3,4,6-five-O-galloyl-β-D-glucose, 1,2,3,4,6-six-galloyl-alpha-D-glucose, 1,2,3,4,6-six-O-galloyl-β-D-glucose, 1,2,3,4,6-seven-O-galloyl-alpha-D-glucose, 1,2,3,4,6-seven-O-galloyl-β-D-glucose, 1,2,3,4,6-eight-O-galloyl-alpha-D-glucose, 1,2,3,4,6-eight-O-galloyl-β-D-glucose, 1,2,3,4,6-nine-O-galloyl-alpha-D-glucose, 1,2,3,4,6-nine-O-galloyl-β-D-glucose, 1,2,3,4,6-ten-O-galloyl-alpha-D-glucose, with 1,2,3,4,6-ten-O-galloyl-β-D-glucose.As used herein, term " fully pure " means the listed gallotannin that the gallotannin compositions comprises a kind of of at least 95% dry weight or a combination, and one or more following chemical compounds that are less than 5% dry weight: list-O-galloyl-β-D-glucose, two-O-galloyl-β-D-glucose, three-O-galloyl-β-D-glucose, four-O-galloyl-β-D-glucose, 11-O-galloyl-β-D-glucose, 12-O-galloyl-β-D-glucose or its mixture.

In another embodiment of this method, comprise to individuality and use a kind of compositions, be called " gallotannin variant compositions " here.The gallotannin variant compositions comprises one or more gallotannin variant compound or its salts.Described gallotannin variant compositions has following structure:

R-X-A( _n)-X-A( _q)-X-A( _z)，

Wherein R is selected from: D-glucose, L-glucose, D-glucose, L-glucose, the D-mannose, L-mannose, D-galactose, the L-galactose, D-allose, L-allose, the D-altrose, L-altrose, D-gulose, L-gulose, D-idose, the L-idose, D-talose, L-talose, D-fructose, L-fructose, alpha-D-xylose, α-D-lyxose, β-D-lyxose, α-D-arabinose, β-D-arabinose, α-D-ribose, β-D-ribose, the D-trehalose, D-maltose, D-cellobiose, inositol, the D-glucitol

X is a kind of ester or ehter bond,

A is a kind of trihydroxybenzoic acid, is selected from: 3,4, and 5-trihydroxybenzoic acid, 2,3,4-trihydroxybenzoic acid, 2,4,6-trihydroxybenzoic acid; Or resorcylic acid, be selected from: 2,3-resorcylic acid, 2,4-resorcylic acid, 3,4-resorcylic acid; Or monohydroxy benzoic acids, be selected from 3-hydroxy benzoic acid and 4-hydroxy benzoic acid,

Wherein, n is 5, and q is 0,1,2,3,4 or 5, and, when R is the D-glucose, the L-glucose, D-mannose, L-mannose, D-galactose, the L-galactose, D-allose, L-allose, D-altrose, the L-altrose, D-gulose, L-gulose, D-idose, L-idose, D-talose, the L-talose, D-fructose, during L-fructose, z is 0;

Wherein, n is 4, and q is 0,1,2,3 or 4, and, when R is: alpha-D-xylose, α-D-lyxose, β-D-lyxose, α-D-arabinose, β-D-arabinose, when α-D-ribose, β-D-ribose, z is 0,1 or 2.

Wherein, n is 6, and q is 0,1,2,3,4,5 or 6, and when R is: when D-glucitol or inositol, z is 0, and

Wherein, n is 8, and q is 0,1,2,3,4,5,6,7 or 8, and when R is the D-trehalose, when D-maltose or D-cellobiose, z is 0.

Every kind of compositions has a kind of structure except following structure in the gallotannin variant compositions: four-O-galloyl-β-D-glucose, 1,2,3,4,6-five-O-galloyl-β-D-glucose, 1,2,3,4,6-six-O-galloyl-β-D-glucose, 1,2,3,4,6-seven-O-galloyl-β-D-glucose, 1,2,3,4,6-eight-O-galloyl-β-D-glucose, 1,2,3,4,6-nine-O-galloyl-β-D-glucose, with 1,2,3,4,6-ten-O-galloyl-β-D-glucose.

In the 3rd embodiment of this method, comprise the combination of using a kind of gallotannin compositions of the present invention and gallotannin variant compositions of the present invention to the patient.

The present invention also provides the method for preventing or treating weight increase in individuality.This method comprises the combination of using gallotannin compositions of the present invention, gallotannin variant compositions of the present invention or gallotannin compositions of the present invention and gallotannin variant compositions of the present invention to individuality.

Adipose cell was divided into the method for adipose cell before the present invention also provided and suppressed.This method comprises the combination of preceding-adipose cell with natural gallotannin compositions of the present invention, gallotannin variant compositions of the present invention or natural gallotannin compositions of the present invention and gallotannin variant compositions of the present invention is contacted.Before described-adipose cell can be in culture or in the mammalian subject body.

The present invention is also about this gallotannin variant compositions, and this chemical compound has structure: R-X-A ( _n)-X-A ( _q)-X-A ( _z), or its salt, and the pharmaceutical composition that comprises this compound or its salt.

Description of drawings

Fig. 1. the structure of five-O-galloyl-D-glucose (PGG).

PGG comprises a glucose core, and is covalently bound by ester bond and five gallates.The carbon 1 of glucose ( ^*) on have two kinds of possible conformations, have two kinds of PGG anomers.The PGG conformation of computer simulation shows that α-PGG is than β-PGG more symmetrical (thereby having less polarity).

The glucose transport stimulating activity of Fig. 2 .PGG and anti--lipogenesis activity.

PGG as mixture or as individual isomer (α or β), is added in adipose cell (A) or the preceding adipose cell (B).GTS activity (A) and the AD activity (B) of PGG are passed through ³The H-glucose uptake is measured.Do not express in the GLUT4 pro-adipose cell, so glucose uptake can be used as the indirect measurement of adipose cell differentiation.

Fig. 3 .PGG is with Kd～10 ^-5M is in conjunction with IR.

3T3-L1 adipose cell in 24 orifice plates respectively with the cold insulin of progressive concentration (at 8pM ¹²⁵The I-insulin exists down) or the cold PGG of progressive concentration (at 20 μ M ¹⁴C-PGG exists down) 4 ℃ of overnight incubation, then, and after removing unconjugated isotope, measurement and cell-bonded ¹²⁵The I-insulin or ¹⁴C-PGG.

Fig. 4. in the 3T3-L1 adipose cell, PGG does not replace insulin and is attached to its receptor IR.

3T3-L1 adipose cell and progressive concentration ¹⁴C-PGG is at 8pM ¹²⁵Under the insulin of I-labelling exists, 4 ℃ of overnight incubation, calculate then ¹²⁵The I-insulin or ¹⁴The cell combination of C-PGG.The standard of PGG-constant insulin is counted between 0.1 and 20 μ M, shows at this concentration range PGG and can not replace insulin from IR.Insulin is in conjunction with raising on higher PGG concentration.

The protein binding selectivity of Fig. 5 .PGG.

In the presence of the epoxy resin or BSA or ovalbumin of variable, fixed amount ¹²⁵I BSA and PGG are hatched jointly.To take place bonded and free the separation by precipitation, compete thing not in the presence of with % in conjunction with expression.Article three, competition curve shows, the PGG selective binding has the different albumen of obvious different affinitys.

Fig. 6. in the 3T3-L1 adipose cell, three kinds of insulin signaling approach-special inhibitor also destroy the inductive glucose transport of PGG-.

Existing or not existing under the different inhibitor, with insulin or PGG induced lipolysis cell.The glucose transport of the cell of handling is active in by cellular uptake ³The H glucose measurement.Be respectively that HNMPA-(AM) 3 suppresses the Tyr kinase activity, cytochalasin (Cytochalasin) B suppresses GLUT4, and wortmannin (Wortmannin) suppresses PI-3K.

Fig. 7. in the 3T3-L1 adipose cell, PGG induces the phosphorylation of Akt.

The adipose cell that the cracking difference is handled is with SDS-PAGE analysis of cells albumen.The non-processing of 1=; The 2=insulin; 3=PGG, 15 μ M; 4=PGG, 30 μ M; The big tick marks of M=albumen.

Fig. 8. add the inductive 3T3-L1 cell of 30 μ M PGG by MDI or insulin with oil red O stain.

After inducing 10 days, the cell oil red O stain is taken pictures under amplifying * 200 times.Only those cells that comprise fat bubble (triglyceride) can be colored.

Fig. 9. before 3T3-L1 in the adipose cell, the Northern engram analysis of PPAR γ and C/EBP α gene expression.Different time after processing, the mRNA that analyzes the preceding adipose cell of difference processing expresses.Road 1: non-processing; Road 2=MDI; Road 3=30 μ M PGG+MDI; Road 4=is the PGG of 30 μ M only.Time=processing after mRNA is separated after the time.

Figure 10. the clonal expression before the 3T3-L1 of α-PGG and β-PGG processing in the adipose cell.

MDI, or the clone of adipose cell expands before existing α-PGG down or β-PGG to induce by MDI.Induced the back 24 or 48 hours, culture medium is removed and cell lysis.Measure lactose dehydrogenase (LDH) activity of cell lysate with the LDH test kit.The LDH activity of while collecting cell growth medium and measurement dead cell (showing in this chart).In given cell type, the LDH activity is constant, and the LDH activity of measurement is proportional with the cell quantity in the sample.

Figure 11. in db/db and ob/ob mice, PGG is for the influence of blood glucose levels.Db/db mice (A) oral administration is not contained the α-PGG of the various dose of glucose, or ob/ob mice (B) oral administration is contained the α-PGG of the various dose of glucose.Different time after administration is measured from the glucose in the blood sample of tail.

Figure 12. PGG protects the ob/ob mice of hyperglycemia immediately behind glucose test, and after test several days hypoglycemia that becomes.The ob/ob mice carries out a kind of glucose tolerance test, shown in Figure 11 B.Different time points behind glucose test is measured blood glucose levels in tail blood.

Figure 13. the chemical constitution of the gallotannin variant of selection.G represents trihydroxybenzoic acid.

Figure 14. in the ob/ob mice, PGG is for the influence of plasma insulin level.

Figure 15. in the mice that has the normal blood glucose level, PGG is for the influence of blood glucose levels.

Detailed description of the present invention

Definition

Term " diabetes " refers to a kind of disease or state, usually to take place in the glucose producing and utilize Metabolic deficiency is feature, and it causes can not keeping suitable blood sugar level in the body. The result of these defectives is blood Portugals Grape sugar raises, and is called " hyperglycemia ". The diabetes of two kinds of principal modes are type 1 diabetes and diabetes B. As previously mentioned, the type 1 diabetes normally absolute shortage of insulin causes, and insulin is to regulate glucose utilization Hormone. Diabetes B often occur in normal or even the situation of the insulin of improving the standard under, by tissue Can not cause by normal response insulin. Most of diabetes B patients are insulin resistances, have relative pancreas The island is plain to be lacked, because insulin secretion can not compensate peripheral tissues to the resistance of insulin replies. And, many 2 Diabetes mellitus type is fat. The glucose homeostasis imbalance of other type comprises the glucose-tolerant of weakening The property, it is a kind of metabolism stage between normal glucose homeostasis and diabetes; And gestational diabetes mellitus, It is not have formerly 1 type and the glucose of diabetes B history in women's pregnancy not anti-.

The glucose tolerance of diagnosed type 2 diabetic, weakening and the policy of gestational diabetes mellitus are by U.S.'s glycosuria Sick association point out (referring to such as diabetes diagnosis and the systematicalian committee, diabetes care, (1999) roll up 2 (Suppl 1): S5-19).

" symptom " of term diabetes includes but not limited to polyuria used herein, polydipsia and voracity, high pancreas The plain mass formed by blood stasis in island and hyperglycemia are in conjunction with they usual usages. Such as, " polyuria " referred in the given time Produce the urine of large volume in phase; " polydipsia " refer to chronically, excessively thirsty; " voracity " refers to eat food excessively Thing, hyperinsulinemia refer to the blood insulin level that improves. Other diabetic symptom comprises as to some infection Neurological susceptibility (especially fungi and staphy lococcus infection) rising, gastric disorder causing nausea and the ketoacidosis (ketoboidies that in blood, strengthens Produce).

" complication " of term glucose includes but not limited to microvascular complication and macrovascular complications. Little blood The pipe complication is those complication that usually cause little blood vessel to damage. These complication comprise as retinopathy (by Cause impaired vision or forfeiture in the damage of eye medium vessels); Neuropathy (causes nerve because neural blood vessel damages Damage and foot problem); And ephrosis (causing kidney trouble because the kidney medium vessels damages). Macrovascular complications is that A little complication that usually caused by the trunk damage. These complication comprise such as heart disease and peripheral artery disease. The heart Vascular diseases refer to the disease of the blood vessel of heart. Referring to such as Kaplan, R.M. etc., " Cardiovascular Diseases " in HEALTH AND HUMAN BEHAVIOR, pp.206-242 (McGraw-Hill, New York 1993). Angiocardiopathy is one of several forms normally, comprise such as hypertension (being also referred to as high vascular pressure), hat Worry, apoplexy and rheumatic heart disease. Peripheral artery disease refers to the disease of the blood vessel of any heart outside, its warp Normal narrow sense is interpreted as carries blood to the blood vessel of leg and arm muscle.

As used herein, " hydrolyzable gallotannin " refers to a kind of galloyl-glucosylation compound, and it is a kind of Portugal The ester of grape sugar is with one or more trihydroxy benzene yl carboxylic acids. Fully pure hydrolyzable gallotannin group of the present invention Compound comprises one or more hydrolyzable gallotannin α or β anomers, has 5,6,7,8,9 or 10 Gallate Acyl group group, or the acceptable salt of its medicine. Six, the hydrolyzable gallotannin of seven, eight, nine and ten forms all has The galloyl group of an initial setting up is attached on the carbon 1,2,3,4 and 6 of glucose core; And secondary The galloyl that arranges is respectively from 1-5 extra galloyl group. The galloyl group of secondary setting is attached to for the first time to be established Put on the galloyl group that separates. Fully pure gallotannin composition of the present invention also can comprise 1,2,3,4-, four-O-Galloyl-alpha-D-glucose.

Many based on the food of plant in the β anomer of hydrolyzable gallotannin, these food comprise light water Really (berry, banana, grape, apple); Cereal (barley, jowar); The beverage of plant origin such as tea and grape wine. Typically, the beta isomer of hydrolyzable gallotannin also can find in the tannic acid mixture, and it is commercial available.

Commercial tannic acid mixture also comprises methyl galloyl and the galloyl glucosylation compound of variable quantity, comprises 1,2,3,11 and 12 galloyl groups. Fully pure hydrolyzable gallotannin composition of the present invention comprises and is lower than 5% and does Heavy, preferably be lower than 3% dry weight, more preferably less than a kind of of the following compound of 1% dry weight or mix: list-O-Gallate Acyl-β-D-Glucose, two-O-galloyl-β-D-Glucose, three-O-galloyl-β-D-Glucose or its mixing. Thereby, The abundant pure hydrolyzable gallotannin composition of using in the present invention is the gallotannin mixture available with commerce Different.

As used herein, " gallotannin variant " refers to a kind of compound, and be similar but be different from following structure: four-O-Galloyl-β-D-Glucose, 1,2,3,4,6-, five-O-galloyl-β-D-Glucose, six-O-galloyl-β-D-Glucose, seven-O-Galloyl-β-D-Glucose, eight-O-galloyl-β-D-Glucose, nine-O-galloyl-β-D-Glucose, ten-O-galloyl-β-D-Glucose, 11-O-galloyl-β-D-Glucose, 12-O-galloyl-β-D-Glucose.

Variant has following structure:

R-X-A( _n)-X-A( _q)-X-A( _z)，

Wherein R is selected from: D-Glucose, L-glucose, D-MANNOSE, the L-mannose, D-galactolipin, L-galactolipin, the D-allose, L-allose, D-altrose, the L-altrose, the D-gulose, the L-gulose, the D-idose, the L-idose, the D-talose, L-talose, D-Fructose, L-fructose, alpha-D-xylose, α-D-lyxose, β-D-lyxose, α-D-R, β-D-R, α-D-ribose, β-D-ribose, D-trehalose, D-Maltose, D-cellobiose, inositol, D-Glucose alcohol

X is a kind of ester or ehter bond,

A is trihydroxybenzoic acid, is selected from: Gallic Acid, 2,3,4-trihydroxybenzoic acid, 2,4,6-Trihydroxybenzoic acid; Or dihydroxy-benzoic acid, be selected from: 2,3-dihydroxy-benzoic acid, 2,4-dihydroxy-benzoic acid, PCA; Or monohydroxy benzoic acids, be selected from 3-hydroxybenzoic acid and 4-HBA,

Wherein, n is that 5, q is 0,1,2,3,4 or 5, and, when R is D-Glucose, L-glucose, D-MANNOSE, the L-mannose, the D-galactolipin, the L-galactolipin, the D-allose, the L-allose, the D-altrose, The L-altrose, D-gulose, L-gulose, D-idose, the L-idose, the D-talose, the L-talose, D-Fructose, during L-fructose, z is 0;

As described here, " adipose cell " is meant adipose cell.In the morphology, adipose cell is circular, comprises the cell of triglyceride (fat) bubble.On the biochemistry, adipose cell is at the high-caliber Insulin receptor INSR of cell surface expression, and represents the glucose transport signal pathway of overactive insulin-mediation, relates to glucose transporter 4 (GLUT4).In vivo, adipose cell relates to the synthetic of fat (triglyceride) and storage, and Fructus Vitis viniferae metabolism (ingestion of glucose from blood, and be converted to fat with glucose).

As used herein, " preceding adipose cell " is meant the adipose cell precursor, under hormone such as insulin and glucocorticoid effect, divides and is divided into adipose cell.In the morphology, preceding adipose cell is fibroblast-sample (thin, and spindle), and lacks triglyceride (fat) bubble in their Cytoplasm.Compare with adipose cell, preceding adipose cell comprises the Insulin receptor INSR and high-caliber relatively insulin like growth factor 1 (IGF-1) receptor of low water level, is used to receive mitogenesis and differentiation signal.Without inducing or all differentiation, preceding adipose cell can not expressed GLUT4 or the relevant gene of other differentiation, as PPAR-γ, C/EBP-α or C/EBP-γ.The glucose transport activity is lower than adipose cell in the cell of preceding adipose cell.

As used herein, " lipogenesis " is meant a process, crosses the Cheng Qian adipose cell separately and be divided into adipose cell by this.

As used herein, " lipogenesis " is meant a process, is synthesized in adipose cell and gathers by this process fat.

Term " mammal " includes but not limited to people, domestic animal (as Canis familiaris L. or cat), farming animals (cattle, horse or pig), monkey, rabbit, mice and laboratory animal.

In one aspect, the invention provides the method at mammalian cell moderate stimulation glucose uptake, the glucose of especially suffering from diabetes, weakening is not anti-, the mammal of insulin resistance or gestational diabetes.In yet another aspect, the invention provides in mammal the method that adipose cell before suppressing is divided into adipose cell, especially fat, overweight or present in the mammal of the not anti-or gestational diabetes of diabetic symptom, glucose.This method part finds that based on the inventor some hydrolyzable gallotannin and some gallotannin variant can stimulate glucose transport to arrive adipose cell, and adipose cell is divided into adipose cell before suppressing.This method also part finds that based on the inventor some hydrolyzable gallotannin can reduce blood glucose levels and blood insulin level in mammal.This method also finds that based on the inventor some hydrolyzable gallotannin does not cause hypoglycemia to small part.Therefore, this method is useful for glucose tolerance, insulin resistance and the gestational diabetes of treatment in mammal and prevent diabetes, weakening.

In one embodiment, the present invention is used for comprising to a kind of abundant pure hydrolyzable gallotannin compositions for the treatment of effective dose of administration in the method for glucose tolerance, insulin resistance and the gestational diabetes of mammal treatment or prevent diabetes, weakening.Fully pure gallotannin compositions comprises one or more hydrolyzable gallotannins, is selected from: 1,2,3, and 4-four-O-galloyl-alpha-D-glucose, 1,2,3,6--four-O-galloyl-alpha-D-glucose, 1,3,4,6-four-O-galloyl-alpha-D-glucose, 1,2,3,4,6-five-O-galloyl-alpha-D-glucose, 1,2,3,4,6-five-O-galloyl-β-D-glucose, 1,2,3,4,6-six-galloyl-alpha-D-glucose, 1,2,3,4,6-six-O-galloyl-β-D-glucose, 1,2,3,4,6-seven-O-galloyl-alpha-D-glucose, 1,2,3,4,6-seven-O-galloyl-β-D-glucose, 1,2,3,4,6-eight-O-galloyl-alpha-D-glucose, 1,2,3,4,6-eight-O-galloyl-β-D-glucose, 1,2,3,4,6-nine-O-galloyl-alpha-D-glucose, 1,2,3,4,6-nine-O-galloyl-β-D-glucose, 1,2,3,4,6-ten-O-galloyl-alpha-D-glucose and 1,2,3,4,6-ten-O-galloyl-β-D-glucose, or the acceptable salt of the pharmacy of hydrolyzable gallotannin.Fully pure hydrolyzable gallotannin compositions comprises one or more following chemical compounds that are less than 5% dry weight: list-O-galloyl-β-D-glucose, two-O-galloyl-β-D-glucose, three-O-galloyl-β-D-glucose, four-O-galloyl-β-D-glucose, 11-O-galloyl-β-D-glucose, 12-O-galloyl-β-D-glucose or its mixture.

In another embodiment, be used for the treatment of or prevent the method for glucose tolerance, insulin resistance and the gestational diabetes of diabetes in the mammal, weakening to comprise and use a kind of gallotannin variant compositions for the treatment of effective dose, wherein comprise one or more gallotannin variant chemical compounds to the patient.In a further embodiment, this method comprises to the patient uses a kind of fully pure hydrolyzable gallotannin compositions and a kind of gallotannin variant compositions of the present invention simultaneously.

Can be randomly, be used to treat or other reagent of prevent diabetes, comprise insulin, sulphur urea, meglitinide class medicine (Meglitinide), biguanides (Biguanide) (Kuruization (Glucophage) or metformin), thiazolidinediones (thiazolidinedione) (TZDs) and Alpha-glucosidase inhibitor, combined with hydrolyzable gallotannin of the present invention or gallotannin variant compositions and delivers medicine to mammal.Suffers from the mammal of type 1 diabetes for those, the conjugate of preferred administration insulin and gallotannin compositions and/or gallotannin variant compositions.

Object

The mammal that this method suffers from the glucose tolerance of diabetes, gestational diabetes, insulin resistance or weakening after diagnosing for treatment is useful.This method also is useful for the mammal that treatment presents the glucose tolerance symptom of diabetes, gestational diabetes, insulin resistance or weakening.This method also is useful for a prevention or a treatment body weight gains, particularly fat or overweight individuality.

Administering mode

This compositions is administered to individuality by injection, comprises subcutaneous, parenteral and intravenous injection, or passes through oral administration.Owing to be easy to administration, preferred route of administration is an oral administration.

Prescription

Use conventional method, gallotannin of using among the present invention and gallotannin variant compositions are made into pharmaceutical composition.These pharmaceutical preparatioies comprise one or more hydrolyzable gallotannins of the present invention and/or one or more gallotannin variant compositions of the present invention.Can be randomly, this pharmaceutical composition also comprises a kind of pharmaceutical acceptable carrier or diluent.Term " pharmacy is acceptable " is meant a kind of atoxic material, can not disturb the effectiveness of gallotannin or gallotannin variant compositions.Many this carriers are conventional uses, and can discern by reference pharmacy books.The characteristic of carrier will depend on specific compound or the combination of compounds in route of administration and the compositions.The preparation of this preparation belongs in those skilled in the art's the horizontal extent.Said preparation can further comprise other and strengthen gallotannin or active or additional its active reagent of gallotannin variant.Said preparation can further comprise implant, salt, cushion, stabilizing agent, solubilizing agent and other material known in the art.

Pharmaceutical composition can exist with unit dosage form easily, and can be by the known method preparation of any pharmaceutical field.Term " unit dose " is meant true quantitative gallotannin of a kind of expection or gallotannin variant compositions, is enough effective in treatment target disease or imbalance.All methods comprise gallotannin compositions, gallotannin variant compositions or both and carrier or diluent and any other selectable supplementary element are contacted.

For oral administration, pharmaceutical composition is such as adopting following form: add tablet or the capsule that the acceptable excipient of pharmacy prepares, excipient such as bonding agent (as corn starch, polyvinyl pyrrolidone or the hydroxypropyl emthylcellulose of pre-gelatinize), filler (as lactose, microcrystalline Cellulose or calcium hydrogen phosphate), lubricant (as magnesium stearate or Talcum), disintegrating agent (as potato starch or carboxymethylstach sodium) or wetting agent (as sodium lauryl sulphate) by conventional instrument.Tablet can wrap quilt by technology well known in the art.Be used for oral liquid preparation such as adopting following form: aqueous solution, syrup or suspension, or can there be water-soluble before use or other suitable media in they by dryed product.This liquid preparation can add that the pharmacy acceptable additive is prepared by the instrument of routine, and additive is such as the media (as almond oil, grease, ethanol or fractionated vegetable oil) of suspending agent (as sorbitol syrups, cellulose derivative or hydrogenant edible fat), emulsifying agent (as lecithin or Radix Acaciae senegalis), non-water and antiseptic (as methyl or propyl group-to hydroxyl (p-hydro) benzoate or sorbic acid).Said preparation can comprise suitable buffer salt, flavoring agent, pigment and sweeting agent.Said preparation also can be taked the mode of nutrient.The preparation that is used for oral administration can be mixed with the gallotannin compositions of sustained release.Owing to have the high-caliber albumen that comprises proline in the saliva, oral formulations is preferably with capsular form, its comprise a skin with protection gallotannin compositions not with ptyaloreaction.

Gallotannin and gallotannin variant compositions can be prepared into the parenteral form by injection, as injecting or continous pouring by pill.The preparation that is used to inject can unit dose form be present in as ampoule bottle or many-dose container, have additional preservatives.The form that compositions can be taked is as the suspension in oil or aqueous medium, solution or Emulsion, and can comprise formula agent as suspend, stable and/or dispersant.Selectively, active component can powder type, tablet or capsule form, and before use with suitable media, as the pyrogen-free water of sterilizing mixes.

Gallotannin and gallotannin variant compositions also can be mixed with rectal compositions, such as suppository or enema, as comprise conventional suppository substrate, as cupu oil, other glyceride or carbon cured (Carbowax).

Except previously described preparation, gallotannin and gallotannin variant compositions also can be made into durative action preparation.This long term preparation can be by implanting (as subcutaneous or intramuscular) or intramuscular injection administration.Therefore, such as, chemical compound can with suitable polymerization or hydrophobic material (such as the Emulsion in the acceptable oil) or ion exchange resin preparation, or as the derivant of indissoluble, as salt as indissoluble.

Dosage

With the treatment effective dose individuality is used gallotannin or gallotannin variant compositions.As used herein, term " treatment effective dose " is meant that total amount is enough to show significant benefit, as the prevention of the improvement of alleviating hyperglycemia (blood sugar level reductions), alleviating hyperinsulinism (reduction of blood insulin level), glucose tolerates, weight increase with lose weight.Need the meaningful result's of acquisition the gallotannin compositions or the dosage of gallotannin variant compositions to carry out routine test with suitable contrast and to determine by this area ordinary person according to the disclosure content.Suitable treatment group and contrast will show that relatively a specific dosage is whether effective for reducing individual blood sugar level or suppressing lipogenesis.

The amount of required gallotannin compositions will depend on the character and the seriousness of the state of an illness of being treated, and the character that depends on individual previous treatment.At last, dosage obtains applying clinical research.At first, the clinicist can use the dosage from zooscopy.A kind of effective dose can reach by the single administration of compositions.Selectively, a kind of effective dose can reach by repeatedly using compositions to individuality.External, biologic effective dose promptly is enough to induce the amount of glucose uptake, by with two multiplication amount administrations, determines active full breadth.Effect oral, subcutaneous and intravenous administration is determined with clinical research.Although single administration gallotannin compositions may be useful, the preferably administration of multidose.

Measure the method for the dosage of glucose uptake in the irritation cell

The glucose uptake activity can be passed through standard test in cell, measures 2-deoxidation-D-[ ³H] picked-up of glucose analyzes.Be grown in the DMEM washed twice of the 3T3-L1 adipose cell that the converges usefulness serum deprivation on 12 orifice plates, the culture medium identical with 1mL hatched 2 hours at 37 ℃.Cell was hatched 30 minutes at 37 ℃ with 0.9ml KRP buffer with Krebs-Ringer-Hepes (KRP) buffer washing 3 times.Then, insulin (positive control) or gallotannin or gallotannin variant (experiment) are added with predetermined concentration, adipose cell was hatched 15 minutes for 37 ℃.Add by 0.1ml KRP buffer and 37MBq/L 2-deoxidation-D-[ ³H] glucose and ultimate density 1mmol/L glucose, the beginning glucose uptake.After 10 minutes,, stop glucose uptake by with cold PBS washed cell 3 times.Cell is with the 1%Triton X-100 of 0.7mL, 37 ℃ of cracking 20 minutes.Determine the radioactivity of cell lysate by scintillation counter.Can in laboratory sample, select to induce the dosage of maximum glucose uptake.

Determine to stimulate the method for the dosage of glucose uptake in the animal

Male db/db (leptin (leptin) receptor deficiency) mice in 8 ages in week can be used to determine the dosage of body internal stimulus glucose uptake.Analyze how much dosage as required, mice is divided into 3 to 4 groups.The test solution that test mice oral administration 10 μ l is had the test gallotannin compositions that pre-determines concentration.The negative control mice is accepted the water of same amount.After administration, different time is collected blood from mousetail behind oral administration.The blood sugar level that the time is gone up mice after the specific administration is singly touched the basis and is measured on the skin treatment or preparation methodologies (available from Lifescan) fully by 6 μ l blood are added in.Effective dosage ranges and optimal dose can be by the contrast various dose the reduction of blood sugar level determine with respect to the blood sugar level of negative (water) matched group.

The inhibition of lipogenesis and weightening finish

In yet another aspect, the invention provides the method that the preceding adipose cell of inhibition is divided into adipose cell.Adipose cell can be in culture or intravital at mammalian subject.In one embodiment, this method comprises preceding adipose cell is contacted with the hydrolyzable gallotannin compositions of biologic effective dose.The gallotannin compositions is fully pure, comprises the gallotannin compositions, comprises following one or more hydrolyzable gallotannins: 1,2,3, and 4-four-O-galloyl-alpha-D-glucose, 1,2,3,6-four-O-galloyl-alpha-D-glucose, 1,3,4,6-four-O-galloyl-alpha-D-glucose, 1,2,3,4,6-five-O-galloyl-alpha-D-glucose, 1,2,3,4,6-five-O-galloyl-β-D-glucose, 1,2,3,4,6-six-galloyl-alpha-D-glucose, 1,2,3,4,6-six-O-galloyl-β-D-glucose, 1,2,3,4,6-seven-O-galloyl-alpha-D-glucose, 1,2,3,4,6-seven-O-galloyl-β-D-glucose, 1,2,3,4,6-eight-O-galloyl-alpha-D-glucose, 1,2,3,4,6-eight-O-galloyl-β-D-glucose, 1,2,3,4,6-nine-O-galloyl-alpha-D-glucose, 1,2,3,4,6-nine-O-galloyl-β-D-glucose, 1,2,3,4,6-ten-O-galloyl-alpha-D-glucose, with 1,2,3,4,6-ten-O-galloyl-β-D-glucose.

In another embodiment, this method comprises preceding adipose cell is contacted with the gallotannin variant compositions that comprises one or more gallotannin variants of biologic effective dose.In another embodiment, this method comprises preceding adipose cell is contacted with hydrolyzable gallotannin compositions of the present invention and gallotannin variant compositions simultaneously.

The step of external definite dosage

For the valid density that is identified for preventing lipogenetic gallotannin or gallotannin variant, with adipose cell before undifferentiated and differentiation-induce cocktail (comprising 3-isobutyl-1-methylxanthine, dexamethasone and insulin (MDI)) or MDI adds gallotannin or the gallotannin variant is hatched external.MDI induces differentiation after about 10 days, and this is apparent, because adipose cell has become adipose cell circular, that comprise the fat bubble before the fibroblast sample.The differentiation degree of cell gathers by the microscopic examination lipid and oil red O stain (vesicle that only comprises triglyceride can be dyed redness), and assesses by the glucose uptake activity that last treated cell of the stage of hatching presents.Select and carry out the glucose uptake test, be used for determining the differentiation degree of adipose cell, this experiment can be by insulin-induced ingestion of glucose based on the adipose cell of observing differentiation, and preceding adipose cell can not.

Determine the method for dosage in vivo

For anti--lipogenesis effect and effective dose in the body of determining gallotannin compositions of the present invention or variant gallotannin compositions, use hereditary diabetes female mice (2 types, the KK-A in 5 ages in week ^y).Gallotannin or gallotannin variant are gone into mice with variable concentrations oral administration every day or peritoneal injection, continue for 6 to 10 weeks.Food intake and the body weight of monitoring mice.When experiment finishes, takes out fatty tissue by the uterus treated and control mice, weigh and contrast.Also take out treated and liver control mice, measure the fat content of liver.Cause maximum reduction of fat content of other fatty tissue in uterus and liver and the dosage of not obvious change food intake is considered to gallotannin of the present invention or the active optimal dose of gallotannin variant compositions lipogenesis resisting.

The typical method for preparing hydrolyzable gallotannin

A. separation method

The hydrolyzable gallotannin of beta form can use HPLC heavily to separate from commercial tannin goods.The HPLC system is Beckman System Gold, is made up of one 125 solvent assembly, 168 PDA detectors and one 508 automatic sampler.In order to separate, use the reverse semi-preparative column of Beckman Ultrasphere C-18 (10.0mm * 250mm internal diameter, 5 μ m).Detect wavelength and be located at 320nm or 330nm.Eluant A is the water that adds 0.1% trifluoroacetic acid, and eluant B is the acetonitrile with 0.1% trifluoroacetic acid.Foxy Jr. fraction collector from ISCO is used to collect each peak in time window.When flow velocity is 3mL/min, permanent solvent gradient A: B reached separation at 82: 18 o'clock in 40 minutes.In these cases, the about retention time of the gallotannin of beta form is as follows:

Five-O-galloyl-β-D-glucose 13.5 minutes.

Six-O-galloyl-β-D-glucose 20.2 minutes.

Eight-O-galloyl-β-D-glucose 22.0 minutes.

Nine-O-galloyl-β-D-glucose 30.0 minutes.

Ten-O-galloyl-β-D-glucose 36.3 minutes.

11-O-galloyl-β-D-glucose 39.8 minutes.

B. synthetic method

The chemical synthesis process of the PGG of α and beta form is made up of four steps:

(i) from the methyl ester of gallate, in acetone, react with chlorotoluene in the presence of potassium carbonate and the potassium iodide, produce the gallate derivant methyl 3,4 of protection, 5-three-O-benzyl gallic acid ester.

(ii) sodium hydroxid cuts ester in ethanol water, forms 3,4,5-three-O-benzyl gallate.

(iii) 3,4,5-three-O-benzyl gallate is used at 1 of 4-(dimethylamino) pyrimidine, and the 2-dichloroethane solution exists down, the D-glucose esterification of dicyclohexylcarbodiimide mediation.Obtain α-and five-O-(3,4,5-three-O-benzyl galloyl)-D-Glucopyranose. of β-anomer form of mixtures.

(iv) in oxolane palladium catalyst in the presence of, hydrogenation five-O-(3,4,5-three-O-benzyl galloyl)-D-Glucopyranose., go the protection.The chromatograph of the blended PGG of α/β (Chromatotron) is separated the independently purified anomer of generation.

The intermediate of natural abundance and the characteristic of product and purity by ¹H and ¹³C{ ¹H}NMR spectrum, electrojet mass spectrum and the visible spectrum of UV-are determined.

Use α-five-O-galloyl glucose, the mixture of β-five-O-galloyl glucose can prepare α-six-O-galloyl glucose as parent material, β-six-O-galloyl glucose, α-seven-O-galloyl glucose, β-seven-O-galloyl glucose, α-eight-O-galloyl glucose, β-eight-O-galloyl glucose, α-nine-O-galloyl glucose, β-nine-O-galloyl glucose, α-ten-O-galloyl glucose, β-ten-O-galloyl glucose.It is identical or closely similar with previously described α/β-synthetic reactions steps of five-O-galloyl glucose that one or more gallates join the reaction of α/β-five-O-galloyl glucose.This reaction will be synthesized the mixture of the galloyl glucose with 6,7,8,9 or 10 gallates.Be separable into single kind by these different chemical compounds of HPLC, and their individual configurations characteristic can be analyzed and verifies by mass spectrum and NMR.

α and β-four-O-galloyl glucose can prepare by an oh group on the protection glucose before adding gallate, and after additive reaction this hydroxyl is gone protection.

The exemplary preparation method of preparation gallotannin variant

1) synthetic five-O-(3,4-dihydroxy benzoyl)-β-D-Glucopyranose .-

Step 1:3,4-benzoic acid dibenzyl ethyl ester

Method:

At room temperature stir 3,4-benzoic acid dibenzyl ethyl ester (10g, 54.3mmol), potassium iodide (4g, 24mmol) and the potassium carbonate of anhydrous powder (40g, 289mmol) mixture in acetone (500mL) is 20 minutes.Add the chlorotoluene be dissolved in the 100mL acetone (14.85g, 117mmol).Suspension returning 18 hours.Cross filter solid, the evaporation filter liquor.Residue is dissolved in the 300mL dichloromethane again, filters once more.Evaporating solvent.Obtain a kind of dark yellow oil.It can be used for next step, without any need for being further purified.

Step 2:3, the 4-benzoic acid dibenzyl

Method:

The crude product of preceding step is suspended in 95% ethanol (300mL), and adding sodium hydroxide precipitation (3.54g, 88.5mmol).Reflux mixture 3 hours.Hot solution is fed in 500mL water and the 25mL concentrated hydrochloric acid.Behind the rotary flask 10 minutes, filtration product, water (100mL), 95% ethanol (100mL), methanol (100mL) and methyl tertiary butyl ether(MTBE) (100mL) washing continuously.White solid is placed oil pump vacuum (～0.1 crust), at room temperature dried overnight.

The output of two steps: 18.6g (93%)

Step 3: five-O-(3,4-benzyloxy benzoyl)-β-D-Glucopyranose.

Method:

With the D-glucose (0.2g, 1.11mmol), 3,4-benzoic acid dibenzyl (2.78g, 8.31mmol), dicyclohexylcarbodiimide (DCC, 1.84,8.92mmol) and N, (DMAP, 1.08g 8.84mmol) join in the dried dichloromethane (130mL) the N-dimethylamino naphthyridine.Suspension returning 2.5 days.Behind cool to room temperature, elimination carbamide by-product.8g silica gel is joined in the filter liquor, be evaporated to drying.Residue is added to (solvent system: dichloromethane: toluene: ethyl acetate=300: 100: 4) on the silicagel column.Merge purified component and evaporation.

The limpid oil of output: 1.72g (88%) high viscosity.

Step 4: five-O-(3,4-dihydroxy benzenes formoxyl)-β-D-Glucopyranose.

Method:

(392mg 0.222mmol) is dissolved among the anhydrous THF (50mL) parent material of benzyl protection.Solution outgased about 30 seconds by a kind of water aspirator, carried out magnetic simultaneously and stirred.Then to the flask argon cleaning.The degassing and flushing are carried out 2 times again.Add 10% palladium carbon (287mg, 0.27mmol).With the mixture degassing, wash with hydrogen then.The degassing and flushing repeat 2 times again.Then in normal pressure hydrogen, under 40 ℃, stirred suspension 5 hours with maximal rate.The mixture cooling, diatomite filtration, evaporation filter liquor.

The amorphous solid foamy of output: 190mg (99%).

Other the gallotannin variant that comprises dihydroxy benzyloxy benzoyl group prepares as previously mentioned, except using the parent material of different protocatechuic acid ethyl esters as step 1.

2) four-O-(3,4-benzyloxy benzoyl)-β-D-synthesizing than glucopyranoside

Step 1:3,4,5-three benzyloxy essence of Niobe

Method:

Stir 3,4 under the room temperature, 5-trihydroxybenzoic acid methyl ester (10g, 54.3mmol), potassium iodide (4g, 24mmol) and the potassium carbonate of anhydrous powder shape (44g, 318mmol) mixture 20 minutes of (500mL) in acetone.Add the chlorotoluene be dissolved in the 100mL acetone (22g, 174mmol).Suspension returning 18h.Cross filter solid, the evaporation filter liquor.Residue places the 400mL dichloromethane.Suspension evaporates filter liquor by diatomite filtration.Residue places the oil pump vacuum, at room temperature dry 45 minutes.

Output: 26.528g (107%) (product comprises some chlorotoluenes, as a kind of impurity)

Step 2:3,4,5-three benzyloxy benzoic acid

Method:

The crude product of abovementioned steps is suspended in 95% ethanol (300mL), and adding sodium hydroxide precipitation (3.54g, 88.5mmol).Mixture heated 3 hours under refluxing.Hot solution is fed in the mixture of 500mL water and 25mL concentrated hydrochloric acid.Behind the rotary flask 10 minutes, filtration product, water (100mL), 95% ethanol (100mL), methanol (100mL) and methyl-tert-butyl ether (100mL) washing continuously.White solid is placed oil pump vacuum (～0.1 crust), at room temperature dried overnight.

The output of two steps: 22.42g (94%)

Step 3: four-O-(3,4,5-three benzyloxy benzoyls)-D-xylopyranose

Method:

The D-xylose (0.2g, 1.33mmol), 3,4,5-three benzyloxy benzoic acid (3.515g, 7.98mmol), dicyclohexylcarbodiimide (DCC, 1.77,8.57mmol) and N, (DMAP, 1.04g 8.49mmol) join in the anhydrous methylene chloride (130mL) the N-dimethylamino naphthyridine.Suspension returning 2.5 days.Behind cool to room temperature, elimination carbamide by-product.9g silica gel is joined in the filter liquor, be evaporated to drying.Residue is added to (solvent system: dichloromethane: toluene: ethyl acetate=300: 100: 4) on the silicagel column.Merge purified component and evaporation.

The beta isomer of output: 208mg

The αYi Gouti of 220mg

The mixed fraction of 770mg (alpha+beta)

Step 4: four-O-(3,4,5-trihydroxy benzene formoxyl)-β-D-Glucopyranose.

Method:

(150mg 0.0815mmol) is dissolved among the anhydrous THF (20mL) parent material of benzyl protection.Solution outgased about 30 seconds by a kind of water aspirator, carried out magnetic simultaneously and stirred.Use the argon cleaning flask then.The degassing and flushing repeat 2 times again.Add 10% palladium carbon (287mg, 0.27mmol).The mixture degassing is washed with hydrogen then.The degassing and flushing repeat 2 times again.Then with suspension in normal pressure hydrogen, stirred 5 hours with maximal rate at 40 ℃.The mixture cooling, diatomite filtration, evaporation filter liquor.

The amorphous solid foamy of output: 62mg (100%).

Other gallotannin variant comprises a kind of sugared core except glucose, and as galactose, mannose, trehalose, maltose, cellobiose, inosite and glucitol, preparation as previously mentioned is except the xylose that adds in step 3 is replaced by another kind of sugar.

3) between gallate and glucose, replace ester bond with ehter bond

Method (the E.Eich that this synthetic first three step is a reported in literature, H.Pertz, M.Kaloga, J.Schulz, M.R.Fesen, A.Mazumder, Y.Pommier, (-)-Arctigenin as a Lead Structure for Inhibitorof Human Immunodeficiency Virus Type-1 Integrase, J.Med.Chem.1996,39,86-95).

The benzyl of the carbohydrate of subsequent step and standard is protected/is gone and protects chemical reaction similar.Last hydrogenesis will be than carbohydrate quickly in conjunction with three oxybenzene methyl groups for benzyl phenol.Can estimate that ehter bond reduces the stability (half-life) that will improve molecule to acid-hydrolyzed sensitivity, thereby improve effect in vivo and comprehensive tangible bioactive persistent period.

Embodiment

Following embodiment only is used for proof for example, and the scope of unrestricted additional claim.The document that all this paper quote is all in this complete introducing.

Embodiment 1: with five-O-galloyl-D-glucose (PGG) at cell moderate stimulation glucose uptake

50: 50 mixture as preceding synthetic α-PGG and β-PGG.As previously mentioned α is separated with the β anomer.β-the PGG of the plant origin that the glucose transport of two kinds of anomers is active and real relatively.The PGG of chemosynthesis is identical on the spectrum with the PGG of real plant origin.

The 3T3-L1 adipose cell is containing the required 10%CO of cell available from ATCC ₂37 ℃ of couveuses in, in adding the DMEM of 10% calf serum, keep, go down to posterity as preceding adipose cell.Induce cocktail by adding MDI, cell is induced to differentiate into adipose cell, as Liu, and F., Kim, J., Li, Y., Liu, X., Li, J.﹠amp; Chen, X. (2001), Flos Caryophylli Lagerstroemia indica L. in the 3T3-L1 cell (Lagerstroemiaspeciosa L.) extract has stimulation of Insulin-Like glucose uptake and adipose cell differentiation-inhibition activity, and J.Nutrition 131:2242-224 is described, and it is incorporated herein with for referencial use.Then not commensurability α-PGG and β-PGG joined in the culture medium, the glucose uptake amount of contrast and treated cell uses the glucose uptake of standard to test, as described in Liu etc.Shown in Fig. 2 A, chemosynthesis have similar glucose transport zest (GTS) with β-PGG plant origin.The GTS activity of α-PGG is generally than the high 10-20% of β-PGG (Fig. 2).

Induce the active mechanism of GTS in order to study PGG, synthetic ¹⁴The PGG of C-labelling also is used for cell research.In order to determine that PGG acts in cell membrane or the cell, give radioactivity PGG and 3T3-L1 adipose cell different time at 37 ℃ or 4 ℃.After hatching, remove excessive radioactivity PGG by washing, collecting cell uses disclosed scheme, is component in cell membrane and the cell with cell separation.The above radioactivity of 90-95% combines with membrane component, proves that PGG acts on the 3T3-L1 cell by being attached to cell membrane.

Use 4 ℃ with ¹⁴The 3T3-L1 cell of C-PGG overnight incubation and the unmarked PGG of incremental change carry out part and replace research.The radioactivity insulin is used as the receptors bind contrast.The test concentration range in, PGG works as part, with target be～10 with apparent receptors bind constant (Kd) ^-5M is in conjunction with (Fig. 3).For GTS and AD activity, what the EC50 that the Kd of estimation is very similar to PGG can not represent.Because PGG has insulinoid GTS activity, and show and to induce the GTS activity with the approach similar that can infer, PGG is by bound insulin receptor-inducible GTS activity in the 3T3-L1 cell to insulin.

Whether have common target in order to measure PGG and insulin, the radioactivity insulin of 3T3-L1 cell and constant density adds the cold PGG of raising amount, or hatches jointly with the cold insulin that radioactivity PGG adds the raising amount.Show that in conjunction with result of the test PGG does not compete the insulin binding site (Fig. 4) that is positioned on the IR with insulin.On the contrary, find that also insulin is not positioned at PGG binding site on the cell membrane with PGG competition.As if thereby PGG combines with IR site except the insulin binding site, perhaps combine with membrane receptor except IR.

The bovine serum albumin of application of radiation labelling (BSA) determines in conjunction with test whether PGG can selective binding albumen by competition as tracer.PGG has nothing in common with each other for the relative affinity of three kinds of standard proteins, differs at least 10 times (Fig. 5), and PGG is higher than 100 times for the different of binding affinity of gel and ovalbumin.This shows that the PGG-albumino reaction has specificity, and the PGG alternative acts on single creature chemistry target spot, as IR.

But the chemical inhibitor of the specificity Profilin/enzyme that relates in the signal pathway of insulin-mediation is used to further identify the molecular target of PGG.Three kinds of inhibitor have been tested: hydroxyl-2-naphthyl methyl phosphonic acids triacetyl oxygen ylmethyl ester (HNMPA-(AM) ₃, a kind of chemical agent (Qiu, Z. wait (2001) J.Biol.Chem.276:11988-11995) that can suppress the IR tyrosine kinase; Wortmannin (wortmannin), a species specificity suppresses the chemical compound (Saperstein, R. wait (1989) Biochemistry 28:5694-5701) of PI-3K; And cytochalasin B, a species specificity suppresses chemical compound (Tomiyama, K. etc. (1995) Biochem.Biophy.Res.Commu.212:263-269 by the glucose transport of GLUT4 mediation; Kletzien, R.F. etc. (1972) J.Biol.Chem.247:2964-2966).All three kinds of inhibitor suppress the GTS activity (Fig. 6) by insulin or PGG mediation fully, prove that all GTS activity by the PGG mediation are by, and the signal pathway by insulin-mediated only.In fact, the inhibitor HNMPA-(AM) of first enzyme in the insulin signaling approach (IR tyrosine kinase) ₃Also can suppress the GTS activity of PGG fully, the molecular target that proves PGG is IR.PGG uses the viewpoint of the GTS approach of insulin-mediation and is further supported by the Western trace, and the Western trace shows that the key protein kinases Akt that comprises in this approach (zz) is by PGG phosphorylation (Fig. 7).

Embodiment 2:PGG is for lipogenetic influence

The 3T3-L1 adipose cell is available from ATCC, keeps and pass through to be preceding adipose cell in the DMEM that adds 10% calf serum, places to contain the required 10%CO of cell ₂37 ℃ of couveuses in.In order to measure PGG for lipogenetic influence, preceding adipose cell and the cocktail of inducing differentiation (MDI, a kind of cocktail that contains 3-isobutyl-1-methylxanthine, dexamethasone (MD)) that comprises 3-isobutyl-1-methylxanthine, dexamethasone (MD) and PGG are hatched jointly; Or add PGG with MDI and hatch jointly.The differentiation (referring to Fig. 2 B) of adipose cell before insulin is replaced by PGG in MDI, and new cocktail can not be induced.These results show that PGG can not replace insulin-induced preceding adipose cell and be divided into adipose cell.

Use cell proliferation test, determine whether PGG suppresses the adipose cell differentiation by retardance clone expansion.This test shows, clone's expansion of the first round not by α-or β-PGG suppress.Second clone's expansion of taking turns is partly suppressed by α-PGG, and is suppressed (Figure 10) fully by β-PGG.Do not understand in clone expansion and the differentiation the different basis between two kinds of anomers.Yet, our result shows, clone's expansion inhibition or apoptosis (programmed cell death) do not cause observed differentiation to suppress in the pro-adipose cell, because only expanding with the clone who limits, α-PGG suppresses to forbid differentiation that comparing with the inductive cell of MDI (data not shown) does not have the obviously cell killing of raising.Therefore, the committed step of inhibition is likely the step that is different from clone's expansion.

In order to study the mechanism that PGG suppresses adipose cell differentiation before the 3T3-L1, we have used gene expression research.This research makes us can determine PGG, and whether specific effect is in the gene that participates in atomization.And gene expression research has overcome the difficulty (Modan-Moses etc. (1998) Biochem.J.333:825-831) of Insulin receptor INSR in the preceding adipose cell (it is at the IR of few 10 times of cell surface expression) research.In order to study the gene that in the AD process, influences, preceding adipose cell and the cocktail of inducing differentiation that comprises 3-isobutyl-1-methylxanthine, dexamethasone and insulin are hatched jointly by PGG; Or add PGG with MDI and hatch jointly.After about 10 days, MDI induces differentiation, because adipose cell becomes circle, comprises the adipose cell of fatty vesicle before the fibroblast sample, variation is apparent (in the middle of Fig. 8).On the contrary, the preceding adipose cell that adds the PGG processing with MDI has kept their fibroblast-like forms, and does not still have fatty vesicle (Fig. 8 right side).

At different time, collect treated cell, separate total RNA, use the complementary probe of the different genes that comprises with atomization to analyze expression by the Northern trace.The Northern engram analysis shows, break up required and by the expression of the inductive gene PPAR-of MDI γ, c/EBP-α by PGG total ban (Fig. 9).Beta-actin level consistent relatively in the cell of differentiation processing shows that it is not (Fig. 9) that is obviously influenced by PGG that other cell processes such as beta-actin are expressed.

Embodiment 3: PGG is to the influence of blood sugar lowering level in diabetic animal

In order to determine whether α-PGG can represent anti--diabetic activity in vivo, with the α-PGG of aqueous solution form to 8 age in week male fasting diabetes db/db mice oral administration.Find to compare with the db/db mice of only accepting vehicle (the identical aqueous solution that does not have α-PGG), the concentration of single dose is that the α-PGG of 25mg/kg body weight obviously reduces the blood sugar level (Figure 11 A) in the db/db mice.The about 15-20% of the reduction of glucose level looks the time (P＜0.01, Figure 11 A) after α-PGG administration and decides.

In order to determine whether α-PGG is effective to improving glucose tolerance in diabetes and obesity mice, uses the ob/ob mice to carry out the glucose tolerance test.Glucose or glucose are added α-PGG be administered orally in male ob/ob mice, different point in time measurement blood sugar level after glucose/PGG administration.Only compare with those, accept the ob/ob mice that glucose adds α-PGG and have obviously low blood sugar level (P＜0.001, Figure 11 B) with the mice that glucose is handled.Interesting is, after α-PGG handles 24 hours, still can observe the blood sugar level of raising, and the influence that proves α-PGG is long lasting relatively.

Because very high-caliber blood sugar level after glucose is tested～24 hours, perhaps because of unusual low glucose level after testing 2～3 days at glucose, the glucose of single high dose can be fatal in the ob/ob mice.We find that the influence of very high glucose level in a short time (Figure 11 B) after α-PGG not only protects the ob/ob mice not tested by glucose also protects mice not to be subjected to the influence (Figure 12) of low-down glucose level after 2-5 days.Although protection mechanism is unknown at present, α-PGG also has some other activity not only with its GTS active protection ob/ob mice probably, the activity relevant as AD-.In these mices, do not find the toxic effect that PGG is relevant.

These zooscopies prove that clearly α-PGG not only acts on the 3T3-L1 cell, and are also effective in diabetes/fat animal model body.

Embodiment 4 gallotannin variants are for the influence of the glucose uptake of cell

We after tested the GTS (table 1) of polymerized polyphenolic in the 3T3-L1 cell system of several types.Flos Caryophylli Lagerstroemia indica L. extract and gallotannin preparation (tannin (a kind of mixture of galloyl glucose); And the PGG of purification) has high activity.Procyanidin has limited activity, and tea catechol EGCG does not have activity.This supports special albumen-PGG reaction to cause the active viewpoint of GTS, because only PGG and closely-related compositions are activated.If the GTS activity only is the common protein bound result of polymerized polyphenolic, procyanidin and the tea catechol goods based on flavonoid (flavonoid) will have activity so.

It is reproducible different in GTS and AD activity to obtain between α-PGG and the β-PGG between (Fig. 2) and PGG and the six-GG (table 1) routinely.May be to cause active different reason between α-PGG of being observed and the β-PGG in the direction of the gallate on the carbon 1 of sugar.A kind of gallotannin variant five galloyl galactose (PGGal) of the present invention and five galloyl deoxynojirimycins (PG-DJM) have been synthesized (Figure 13).We find that PGGal has the activity of the 60-70% of PGG, and PG-DJM does not have the GTS activity that can survey.

Glucose transport stimulating activity-β-D-the pyrans of table .PGG derivant

The GTS activity of α-PGG is ++ +++, the activity of β-PGG is ++ ++.

Embodiment 5:PGG is for the influence of plasma insulin level

Injection water or α-PGG in ob/ob mouse peritoneum diabetes and obesity.Different time separates the blood plasma from every mice after injection, measures insulin level.As shown in figure 15, the ob/ob diabetes of single injection α-PGG are compared with the ob/ob diabetic mice (negative control) of only using water treatment with obesity mice and are had significantly low plasma insulin level.To the influence of glucose uptake, can believe that in vivo, PGG can strengthen the glucose transport stimulating activity of insulin based on these results and PGG.Therefore, expection PGG treatability in mammalian subject is used to improve insulin resistance.

Embodiment 6 five-O-(3,4-dihydroxy benzenes formoxyl)-B-D-Glucopyranose. and four-O-(3,4, the 5-trihydroxy The xylopyranose of benzoyl-a-D) is for the influence of the glucose uptake of adipose cell

Be grown in the DMEM washed twice of the 3T3-L1 adipose cell that the converges usefulness serum deprivation on 12 orifice plates, the culture medium identical with 1mL hatched 2 hours at 37 ℃.Cell was hatched 30 minutes at 37 ℃ with 0.9ml KRP buffer with Krebs-Ringer-Hepes (KRP) buffer washing 3 times.Then, be listed in chemical compound in the following table with 20-40 μ M (ultimate density?) add, and adipose cell was hatched 15 minutes at 37 ℃.By adding 0.1mL KRP buffer and 37MBq/L 2-deoxidation-D-[ ³H] glucose and 1mmol/L glucose be as ultimate density, starts the picked-up of glucose.After 10 minutes,, stop glucose uptake in cold PBS by cell is washed 3 times.With the 1%Triton X-100 dissolving of cell, carried out 20 minutes at 37 ℃ with 0.7mL.Determine by scintillation counter by the radioactivity that the cytolysis thing keeps.As shown in Table, gallotannin variant five-O-(3,4-dihydroxy benzoyl)-β-D-Glucopyranose. and four-O-(3,4,5-trihydroxy benzoyl)-α-D-xylopyranose have improved the glucose uptake of cell.

Embodiment 7:PGG does not cause hypoglycemia

Normally the mice of (health) is at the glucose of oral 40mg of 0 time.In 2 hours (120 minutes) back but blood glucose during in normal or foundation level, the PGG of the oral variable concentrations of mice or water (negative control), or peritoneal injection insulin.At the different time of administration PGG or insulin at interval, collect to determine blood sugar level from the blood of contrast and treated animal.As shown in figure 15, injection of insulin causes hypoglycemia in mice, do not have and use PGG.Therefore, PGG reduces than normal high blood glucose levels.Yet PGG can further not reduce the blood sugar level that is lower than normal or basic horizontal.

Claims

1. the treatment or the method for prevent diabetes in a mammalian subject comprise: to a kind of gallotannin compositions for the treatment of effective dose of the individual administration of needs,

Wherein, described gallotannin compositions comprises one or more hydrolyzable gallotannins, is selected from: 1,2,3,4-four-O-galloyl-alpha-D-glucose, 1,2,3,6--four-O-galloyl-alpha-D-glucose, 1,3,4,6-four-O-galloyl-alpha-D-glucose, 1,2,3,4,6-five-O-galloyl-alpha-D-glucose, 1,2,3,4,6-five-O-galloyl-β-D-glucose, 1,2,3,4,6-six-galloyl-alpha-D-glucose, 1,2,3,4,6-six-O-galloyl-β-D-glucose, 1,2,3,4,6-seven-O-galloyl-alpha-D-glucose, 1,2,3,4,6-seven-O-galloyl-β-D-glucose, 1,2,3,4,6-eight-O-galloyl-alpha-D-glucose, 1,2,3,4,6-eight-O-galloyl-β-D-glucose, 1,2,3,4,6-nine-O-galloyl-alpha-D-glucose, 1,2,3,4,6-nine-O-galloyl-β-D-glucose, 1,2,3,4,6-ten-O-galloyl-alpha-D-glucose, with 1,2,3,4,6-ten-O-galloyl-β-D-glucose or its salt, and

Wherein, described gallotannin compositions comprises one or more following chemical compounds that are less than 5% dry weight: list-O-galloyl-β-D-glucose, two-O-galloyl-β-D-glucose, three-O-galloyl-β-D-glucose, four-O-galloyl-β-D-glucose, 11-O-galloyl-β-D-glucose, 12-O-galloyl-β-D-glucose or its mixture.

2. the process of claim 1 wherein that described gallotannin compositions comprises 1,2 of at least 50% dry weight, 3,4,6-five-O-galloyl-alpha-D-glucose or 1,2,3,4,6-five-O-galloyl-β-D-glucose, or 1,2,3,4,6-five-O-galloyl-alpha-D-glucose and 1,2,3,4, the conjugate of 6-five-O-galloyl-β-D-glucose.

3. the process of claim 1 wherein that described gallotannin compositions comprises 1,2,3,4 of at least 50% dry weight, 6-five-O-galloyl-alpha-D-glucose.

4. the process of claim 1 wherein that described gallotannin compositions comprises 1,2,3,4 of at least 50% dry weight, 6-five-O-galloyl-β-D-glucose.

5. the process of claim 1 wherein that described gallotannin compositions is applied to individuality by oral or injection.

6. the method for claim 1 further comprises the step to individual administration of insulin.

7. the treatment or the method for prevent diabetes in an individuality comprise:

Use the gallotannin variant compositions to individuality,

Wherein, described gallotannin variant compositions comprises one or more gallotannin variant compound or its salts, and wherein every kind of described gallotannin variant chemical compound has following structure:

R-X-A( _n)-X-A( _q)-X-A( _z)，

Wherein R is selected from: the D-glucose, the L-glucose, the D-mannose, the L-mannose, the D-galactose, the L-galactose, the D-allose, the L-allose, the D-altrose, the L-altrose, the D-gulose, the L-gulose, the D-idose, the L-idose, the D-talose, the L-talose, D-fructose, L-fructose, alpha-D-xylose, α-D-lyxose, β-D-lyxose, α-D-arabinose, β-D-arabinose, α-D-ribose, β-D-ribose, the D-trehalose, D-maltose, the D-cellobiose, inositol, the D-glucitol

X is a kind of ester or ehter bond,

A is a trihydroxybenzoic acid, is selected from: 3,4, and 5-trihydroxybenzoic acid, 2,3,4-trihydroxybenzoic acid, 2,4,6-trihydroxybenzoic acid; Or resorcylic acid, be selected from: 2,3-resorcylic acid, 2,4-resorcylic acid, 3,4-resorcylic acid; Or the monohydroxy benzoic acid, be selected from 3-hydroxy benzoic acid and 4-hydroxy benzoic acid,

N is 5, q is 0,1,2,3,4 or 5, and, when R was D-glucose, L-glucose, D-mannose, L-mannose, D-galactose, L-galactose, D-allose, L-allose, D-altrose, L-altrose, D-gulose, L-gulose, D-idose, L-idose, D-talose, L-talose, D-fructose, L-fructose, z was 0;

N is 4, and q is 0,1,2,3 or 4, and when R is: when alpha-D-xylose, α-D-lyxose, β-D-lyxose, α-D-arabinose, β-D-arabinose, α-D-ribose, β-D-ribose, z is 0,1 or 2.

N is 6, and q is 0,1,2,3,4,5 or 6, and when R is: when D-glucitol or inositol, z is 0, and

N is 8, and q is 0,1,2,3,4,5,6,7 or 8, and when R was D-trehalose, D-maltose or D-cellobiose, z was 0.

8. the method for claim 7, wherein, X is a kind of ehter bond.

9. the method for claim 7, wherein, R is L-glucose, D-mannose, L-mannose, D-galactose, L-galactose, D-allose, L-allose, D-altrose, L-altrose, D-gulose, L-gulose, D-idose, L-idose, D-talose, L-talose, D-fructose, L-fructose, alpha-D-xylose, α-D-lyxose, β-D-lyxose, α-D-arabinose, β-D-arabinose, α-D-ribose, β-D-ribose, D-trehalose, D-maltose, D-cellobiose, inositol or D-glucitol.

10. the method for claim 4, wherein, A is a kind of trihydroxybenzoic acid, is selected from: 2,3,4-trihydroxybenzoic acid, 2,4,6-trihydroxybenzoic acid; Or a kind of resorcylic acid, be selected from: 2,3-resorcylic acid, 2,4-resorcylic acid, 3,4-resorcylic acid; Or a kind of monohydroxy benzoic acid, be selected from 3-hydroxy benzoic acid and 4-hydroxy benzoic acid.

11. the method for claim 4, wherein, every kind of described gallotannin variant chemical compound has the structure except following structure: four-O-galloyl-β-D-glucose, 1,2,3,4,6-five-O-galloyl-β-D-glucose, 1,2,3,4,6-six-O-galloyl-β-D-glucose, 1,2,3,4,6-seven-O-galloyl-β-D-glucose, 1,2,3,4,6-eight-O-galloyl-β-D-glucose, 1,2,3,4,6-nine-O-galloyl-β-D-glucose, with 1,2,3,4,6-ten-O-galloyl-β-D-glucose.

A 12. blood glucose levels that in required individuality, reduce to improve and do not cause and comprise the method for hypoglycemia:

Use a kind of or use following compositions (a) simultaneously and (b) to individuality,

(a) a kind of gallotannin compositions comprises one or more hydrolyzable gallotannins, is selected from 1,2,3,4-four-O-galloyl-alpha-D-glucose, 1,2,3,6--four-O-galloyl-alpha-D-glucose, 1,3,4,6-four-O-galloyl-alpha-D-glucose, 1,2,3,4,6-five-O-galloyl-alpha-D-glucose, 1,2,3,4,6-five-O-galloyl-β-D-glucose, 1,2,3,4,6-six-galloyl-alpha-D-glucose, 1,2,3,4,6-six-O-galloyl-β-D-glucose, 1,2,3,4,6-seven-O-galloyl-alpha-D-glucose, 1,2,3,4,6-seven-O-galloyl-β-D-glucose, 1,2,3,4,6-eight-O-galloyl-alpha-D-glucose, 1,2,3,4,6-eight-O-galloyl-β-D-glucose, 1,2,3,4,6-nine-O-galloyl-alpha-D-glucose, 1,2,3,4,6-nine-O-galloyl-β-D-glucose, 1,2,3,4,6-ten-O-galloyl-alpha-D-glucose and 1,2,3,4,6-ten-O-galloyl-β-D-glucose or the acceptable salt of its pharmacy, and

(b) a kind of gallotannin compositions comprises the chemical compound that one or more have following structure, or the acceptable salt of its pharmacy:

R-X-A( _n)-X-A( _q)-X-A( _z)，

Wherein R is selected from: D-glucose, L-glucose, D-mannose, L-mannose, D-galactose, the L-galactose, D-allose, L-allose, D-altrose, L-altrose, D-gulose, L-gulose, D-idose, L-idose, D-talose, L-talose, D-fructose, L-fructose, alpha-D-xylose, α-D-lyxose, β-D-lyxose, α-D-arabinose, β-D-arabinose, α-D-ribose, β-D-ribose, D-trehalose, D-maltose, the D-cellobiose, inositol, D-glucitol

X is a kind of ester or ehter bond,

A is a trihydroxybenzoic acid, is selected from: 3,4, and 5-trihydroxybenzoic acid, 2,3,4-trihydroxybenzoic acid, 2,4,6-trihydroxybenzoic acid; Or resorcylic acid, be selected from: 2,3-resorcylic acid, 2,4-resorcylic acid, 3,4-resorcylic acid; Or monohydroxy benzoic acids, be selected from 3-hydroxy benzoic acid and 4-hydroxy benzoic acid,

N is 5, and q is 0,1,2,3,4 or 5, and, when R is the D-glucose, the L-glucose, D-mannose, L-mannose, D-galactose, the L-galactose, D-allose, L-allose, D-altrose, the L-altrose, D-gulose, L-gulose, D-idose, L-idose, D-talose, the L-talose, D-fructose, during L-fructose, z is 0;

N is 4, and q is 0,1,2,3 or 4, and, when R is: alpha-D-xylose, α-D-lyxose, β-D-lyxose, α-D-arabinose, β-D-arabinose, when α-D-ribose, β-D-ribose, z is 0,1 or 2.

N is 8, and q is 0,1,2,3,4,5,6,7 or 8, and when R is the D-trehalose, when D-maltose or D-cellobiose, z is 0.

13. a treatment presents the method for the individuality of the not anti-disease of one or more obesities, type 2 diabetes mellitus, glucose, insulin resistance, hyperglycemia or Hyperinsulinism, comprises

Use a kind of gallotannin compositions to individuality,

Wherein said gallotannin compositions comprises one or more hydrolyzable gallotannins, is selected from 1,2,3,4-four-O-galloyl-alpha-D-glucose, 1,2,3,6--four-O-galloyl-alpha-D-glucose, 1,3,4,6-four-O-galloyl-alpha-D-glucose, 1,2,3,4,6-five-O-galloyl-alpha-D-glucose, 1,2,3,4,6-five-O-galloyl-β-D-glucose, 1,2,3,4,6-six-galloyl-alpha-D-glucose, 1,2,3,4,6-six-O-galloyl-β-D-glucose, 1,2,3,4,6-seven-O-galloyl-alpha-D-glucose, 1,2,3,4,6-seven-O-galloyl-β-D-glucose, 1,2,3,4,6-eight-O-galloyl-alpha-D-glucose, 1,2,3,4,6-eight-O-galloyl-β-D-glucose, 1,2,3,4,6-nine-O-galloyl-alpha-D-glucose, 1,2,3,4,6-nine-O-galloyl-β-D-glucose, 1,2,3,4,6-ten-O-galloyl-alpha-D-glucose and 1,2,3,4,6-ten-O-galloyl-β-D-glucose or its salt, and

14. the method for claim 13 wherein, is used described gallotannin compositions and suppress the differentiation of preceding adipose cell to adipose cell in individuality.

15. the method for claim 13, wherein, described gallotannin compositions by oral or the injection to individual administration.

16. one kind is suppressed the method that preceding adipose cell is divided into adipose cell, comprises in external or body:

Adipose cell contacts with the gallotannin compositions before making,

Wherein, described gallotannin compositions comprises one or more hydrolyzable gallotannins, is selected from 1,2,3,4-four-O-galloyl-alpha-D-glucose, 1,2,3,6--four-O-galloyl-alpha-D-glucose, 1,3,4,6-four-O-galloyl-alpha-D-glucose, 1,2,3,4,6-five-O-galloyl-alpha-D-glucose, 1,2,3,4,6-five-O-galloyl-β-D-glucose, 1,2,3,4,6-six-galloyl-alpha-D-glucose, 1,2,3,4,6-six-O-galloyl-β-D-glucose, 1,2,3,4,6-seven-O-galloyl-alpha-D-glucose, 1,2,3,4,6-seven-O-galloyl-β-D-glucose, 1,2,3,4,6-eight-O-galloyl-alpha-D-glucose, 1,2,3,4,6-eight-O-galloyl-β-D-glucose, 1,2,3,4,6-nine-O-galloyl-alpha-D-glucose, 1,2,3,4,6-nine-O-galloyl-β-D-glucose, 1,2,3,4,6-ten-O-galloyl-alpha-D-glucose, with 1,2,3,4,6-ten-O-galloyl-β-D-glucose or its salt.

17. the method for claim 16, wherein, described gallotannin compositions comprises one or more following chemical compounds that are less than 5% dry weight: list-O-galloyl-β-D-glucose, two-O-galloyl-β-D-glucose, three-O-galloyl-β-D-glucose, four-O-galloyl-β-D-glucose, 11-O-galloyl-β-D-glucose, 12-O-galloyl-β-D-glucose or its mixture.

18. the method for claim 16, wherein, described gallotannin compositions comprises 1 of at least 50% dry weight, 2,3,4,6-five-O-galloyl-alpha-D-glucose or 1,2,3,4,6-five-O-galloyl-β-D-glucose, or galloyl-alpha-D-glucose and 1,2,3,4, the conjugate of 6-five-O-galloyl-β-D-glucose.

19. the method for claim 16, wherein, described gallotannin compositions comprises 1,2,3,4 of at least 50% dry weight, 6-five-O-galloyl-β-D-glucose.

20. one kind is suppressed the method that preceding adipose cell is divided into adipose cell, comprises in external or body

With preceding adipose cell with comprise one or more compositionss and contact with compound or its salt of following structure:

R-X-A( _n)-X-A( _q)-X-A( _z)，

Wherein R is selected from: D-glucose, L-glucose, D-mannose, L-mannose, D-galactose, the L-galactose, D-allose, L-allose, D-altrose, L-altrose, D-gulose, L-gulose, D-idose, the L-idose, D-talose, L-talose, D-fructose, L-fructose, alpha-D-xylose, α-D-lyxose, β-D-lyxose, α-D-arabinose, β-D-arabinose, α-D-ribose, β-D-ribose, D-trehalose, D-maltose, the D-cellobiose, inositol, D-glucitol

X is a kind of ester or ehter bond,

21. a gallotannin variant compositions comprises one or more gallotannin variant chemical compounds or the acceptable salt of its pharmacy, wherein, every kind of described gallotannin variant chemical compound has following structure:

R-X-A( _n)-X-A( _q)-X-A( _z)，

X is a kind of ester or ehter bond,

N is 8, and q is 0,1,2,3,4,5,6,7 or 8, and when R is the D-trehalose, when D-maltose or D-cellobiose, z is 0,

And

Wherein, every kind of β isomeric form that described gallotannin variant chemical compound is not following chemical compound: four-O-galloyl-D-glucose, five-O-galloyl-D-glucose, six-O-galloyl-alpha-D-glucose, seven-O-galloyl-D-glucose, eight-O-galloyl-D-glucose, nine-O-galloyl-D-glucose and ten-O-galloyl-D-glucose.

22. the compositions of claim 21, wherein, described compositions comprises five-O-(3,4-dihydroxy benzoyl)-β-D-Glucopyranose. four-O-(3,4,5-trihydroxy benzoyl)-α-D-xylopyranose.

23. glucose tolerance, hyperglycemia, Hyperinsulinism or a fat compositions that is used for treating at individuality diabetes, insulin resistance, weakening, described compositions has following structural formula:

R-X-A( _n)-X-A( _q)-X-A( _z)，

X is a kind of ester or ehter bond,

Or the acceptable salt of the pharmacy of described chemical compound.

24. glucose tolerance, hyperglycemia, Hyperinsulinism or a fat pharmaceutical composition that is used for treating at individuality insulin resistance, weakening, described pharmaceutical composition comprises chemical compound or the acceptable salt of its pharmacy and the pharmaceutically acceptable carrier or the diluent of the claim 23 of effective dose.