CN103709157B - Poly-nuclear molecular compound and preparation method and application thereof - Google Patents

Poly-nuclear molecular compound and preparation method and application thereof Download PDF

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CN103709157B
CN103709157B CN201410015130.2A CN201410015130A CN103709157B CN 103709157 B CN103709157 B CN 103709157B CN 201410015130 A CN201410015130 A CN 201410015130A CN 103709157 B CN103709157 B CN 103709157B
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molecular compound
berberrubine
methanol
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CN103709157A (en
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李卫民
袁晓
荣向路
周东斌
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Pi & Pi Technology Inc
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    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D455/00—Heterocyclic compounds containing quinolizine ring systems, e.g. emetine alkaloids, protoberberine; Alkylenedioxy derivatives of dibenzo [a, g] quinolizines, e.g. berberine
    • C07D455/03—Heterocyclic compounds containing quinolizine ring systems, e.g. emetine alkaloids, protoberberine; Alkylenedioxy derivatives of dibenzo [a, g] quinolizines, e.g. berberine containing quinolizine ring systems directly condensed with at least one six-membered carbocyclic ring, e.g. protoberberine; Alkylenedioxy derivatives of dibenzo [a, g] quinolizines, e.g. berberine
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
    • C07D491/12—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains three hetero rings
    • C07D491/14—Ortho-condensed systems
    • C07D491/147—Ortho-condensed systems the condensed system containing one ring with oxygen as ring hetero atom and two rings with nitrogen as ring hetero atom
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    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
    • A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics

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Abstract

The invention discloses a poly-nuclear molecular compound. The poly-nuclear molecular compound is prepared by synthesizing berberrubine and magn olol. A brand new poly-nuclear molecular compound is provided for the first time. The poly-nuclear molecular compound is prepared by synthesizing berberrubine and magnolol, and the medicament metabolism loops of absorption, distribution, metabolism and excretion of a single component or a mixture of two components are changed. Compared with berberrubine and magnolol, the obtained poly-nuclear molecular compound has the advantages of remarkably lowered toxicity being only 10 percent that of berberrubine and magnolol, remarkable increase in absorption rate, higher bioavailability, longer metabolism time, long-acting effect and enhanced blood sugar reducing effect. Moreover, the invention further discloses a preparation method of the poly-nuclear molecular compound, and application of the compound to preparation of diabetes drugs.

Description

A kind of multinuclear molecular compound, Preparation Method And The Use
Technical field
The invention belongs to pharmaceutical chemistry and pharmacotherapeutics field, be specifically related to a kind ofly synthesize by berberrubine and magnolol the multinuclear molecular compound, its preparation method that obtain and preparing the purposes in the medicine for the treatment of diabetes.
Background technology
Diabetes are a kind of chronic diseases, when pancreas does not produce enough Regular Insulin or human body cannot effectively utilize produced Regular Insulin, just there will be diabetes.Regular Insulin is a kind of hormone regulating blood sugar.Hyperglycemia or blood sugar increasing, be the out-of-control a kind of normally results of diabetes, the time one can bring grievous injury to many systems of human body for a long time, particularly neural and blood vessel.Diabetes are generally divided into insulin-dependent diabetes mellitus (IDDM), type II diabetes and gestational diabetes.
Impaired glucose tolerance (IGT) and impaired fasting glucose (IFG) refer to a kind of intermediateness of blood sugar for human body value transitory stage between normal and diabetes glucose value.Impaired glucose tolerance patient or impaired fasting glucose patient face the high risk developing into type II diabetes, although this is not inevitable.
Diabetes de-velopment goes down, and may damage heart, blood vessel, eyes, kidney and nerve.
Diabetes are one of most important Chronic Non-Communicable Diseasess of current threat global human health.In order to assess the impact of diabetes on the whole world, the policy formulated for diabetes for international community and national governments and medical institutions provides foundation, and IDF (IDF) is regularly according to the latest data regular update " IDF diabetes map " of global diabetes epidemiology and health economics research.In the vital document of this part of IDF, IDF not only provides global disease condition and the economical load of current up-to-date diabetes, also predicts the incidence trend of global diabetes from now on.On November 14th, 2013, IDF formally discloses sixth version " IDF diabetes map " when borrowing " United Nations's diabetes day " global concern diabetes.
According to the recent statistics of IDF (IDF), 2013 global diabetes 20-79 year adult in morbidity be 8.3%, patient numbers has reached 3.82 hundred million, and wherein 80% in medium and low income country, and in these countries in zooming trend.Estimate that the whole world in 2035 will have nearly 5.92 hundred million people to suffer from diabetes.Currently suffering from the crowd of diabetes, having 1.75 hundred million (46%) not diagnosed.Within 2013, pregnant woman has the number of hyperglycemia to be 2,100 ten thousand, accounts for all puerpera's then 17%.The whole world in 2013 has 79000 new type 1 diabetes patients and is diagnosed.
2013 global impaired glucose tolerance (IGT) morbidity be 6.9%, have 3.16 hundred million people.Expecting 2035 will have 4.71 hundred million people to suffer from impaired glucose tolerance.
The whole world in 2013 has 5,100,000 people and dies from the disease relevant to diabetes, accounts for 8.39% of all death tolls.The global medical cost of this year diabetes reaches 5,480 hundred million dollars, accounts for 11% of global medical expenditure.Expect 2035, the global medical cost relevant to diabetes will reach 6,273 hundred million dollars.The quick growth of diabetes in China and other developing countries, brings very white elephant to the social and economic development of these countries.
In the estimation to every country and regional sickness rate and incidence trend, the number of patients of China's diabetes in 2013 is 9,840 ten thousand, occupying the whole world the first, is secondly India (6,510 ten thousand), the U.S. (2,440 ten thousand), Brazil (1,190 ten thousand), Russia (1,090 ten thousand).Other numbers of patients are lower than 1,000 ten thousand but the country coming top ten also comprises Mexico, Indonesia, Germany, Egypt and Japanese.IDF estimates, the diabetes number of patients to China in 2035 will reach 1.43 hundred million, still occupy the whole world the first, and the U.S. only will reach 2,970 ten thousand.
In this part of vital document, IDF reminds common people again: diabetes are huge and global problems constantly increased the weight of, and diabetes bring more and more white elephant to society; In all countries, the people being in social low layer easily suffers from diabetes all the better and bears relatively heavier burden.Diabetes have been not only a simple health problem, and solving the problem of diabetes needs policy that social many aspects are concrete and action.
The brief introduction of type II diabetes conventional animal model:
(1) spontaneous genetic model:
Ob/ob mouse: this mouse bulimia, obesity, and suffer from diabetes, its obesity causes because ob gene (ob) there occurs recessive mutation, ob/ob mouse of therefore gaining the name (obese mice).Ob/ob mouse also exists exception and the phenotype change of genetic background simultaneously, and wherein C57B/6J system exception is modal genetic background.
Db/db mouse: db/db mouse (diabetic mice), the rice chromosome of this kind of model has pernicious sudden change, and male and female mouse all diabetes occurs.Namely plasma insulin level started to rise from 2 ~ 3 week age, and islet cells also has the changes such as hypertrophy, propagation, retting conditions, and many foods, obesity also clearly, have significant insulin resistant in its fat hyperglycemia phase.
KK with KK-Ay mouse: KK mouse has the character similar with adult's non-insulin-dependent diabetes mellitus, belongs to congenital heredity defective mouse.The feature of obvious obesity, diuresis can be there is during 16 week age, and start to occur hyperglycemia, hyperinsulinemia and blood fat disorder.
STZ-SHR and SHROB rat: give streptozotocin (STZ) process at introduction stage by spontaneous hypertensive rat (SHR), merges essential hypertension model (STZSHR) to just obtaining type II diabetes time adult.Fat spontaneous hypertensive rat (SHROB) and the difference of thin SHR are that SHROB superposes the fat mouse kind showed in hypertensive genetic background, there is high-caliber fasting plasma hyperglycemic-glycogenolytic factor and free fatty acids, and to glucose load abnormal reaction.
Zucker obese rat: Zucker obese rat is the homozygotic individual (fa/fa) of genetic flaw, with hyperglycemia, hyperinsulinemia, hyperlipidemia and moderate hypertension for feature.
OLETF rat: be the Spontaneous type Ⅱ diabetes mouse kind developed by Japanese pharmaceutical companies for 1984.The expression of this mouse cholecystokinin (CCK)-A receptor mrna lacks completely, ODB1 with the ODB2 gene that it carries is relevant with the morbidity of diabetes.This mouse plays the obvious insulin resistant of appearance 12 week age, can be used for the evaluation of insulin resistant intervening measure.
JCR LA-cp rat: be metabolism syndrome (MS) model turning to performance characteristic with Atherosclerosis.Recessive cp genetic homozygous (cp/cp) of androautosome often shows as obesity, insulin resistant, hyperinsulinemia and hypertriglyceridemia etc., has the characteristic feature of MS.
Wistar obese rat: the Wistar-Kyoto rat with impaired glucose tolerance carries the fat gene of fa, and has hyperglycemia and leptin receptor abnormality.
WOKW rat: Wistar Ottawa Karlsburg W (WOKW) rat shows as obesity, moderate hypertension, hyperlipemia, hyperinsulinemia and impaired glucose tolerance and proteinuria, very similar to mankind MS, can be used for the research that human insulin resists relative disease.It affects body weight and the genetic locus of causeing fat is positioned on No. 1 and No. 5 karyomit(e)s.
GK rat: GK rat is a kind of Spontaneous type Ⅱ diabetes model, has the features such as the insulin secreting ability that blood sugar slightly raises, glucose stimulates is impaired, the minimizing of B cells of pancreas agglomerate amount.
(2) genetic modification model
A-ZIP/F-1 with aP2-SREBP-1c transgenic mice: A-ZIP/F-1 transgenic mice and aP2-SREBP-1c transgenic mice are very similar in phenotype, and occur mild fatty liver during birth, blood triglyceride levels raises gradually subsequently.Due to fatty tissue dysplasia, usually there is a series of MS diseases such as the rising of hyperglycemia, hyperinsulinemia, hypertriglyceridemia, free fatty acid levels and insulin resistant in this type of mouse.
TKO-OBR mouse: because leptin level of adipose tissue opposing in periphery also can be causeed fat, technology acquisition TKO-OBR mouse can be knocked out by applying gene, this Mouse Liver and skeletal muscle triglyceride levels raise 4 times and 2 times respectively, and occur impaired glucose tolerance and hyperinsulinemia.
Insulin receptor gene knocks out model: mouse is at insulin receptor (insulin receptor, INSR) during genetic flaw, slight growth retardation is only had during birth, not metabolic disturbance symptom, but there is ketoacidosis very soon after birth, after several days, B cell nonfunction, then dies from ketoacidosis.Therefore, the deratization of INSR clpp gene respectively cannot organize adult rats in the function of INSR analyze.
(3) diet induced model
STZ+ height fat feeds model: C57BL/6J mouse peritoneal injection low dosage STZ, slight damage B cell, feeds and raise high lipid food, can copy the animal model closely similar with clinical type II diabetes after stable.This is a very ripe animal model of diabetes mellitus type II.
SD rat: 8 week age, SD rat was fed after certain hour through high lipid food, shows as obvious hyperinsulinemia, body weight increases and visceral fat accumulation, and prompting rat exists insulin resistant.
Hamster: the rodent with hyperlipemia mostly is rabbit and hamster, and need hypercholesterolemia or higher fatty acid nursing induction preparation.
Gottingen piggy: with higher fatty acid full diet induction 9 ~ 10 month female Gottingen piggys, after 5 weeks, in abdomen, lipid content and blood triglyceride levels obviously raise, and its metabolism is impaired similar to human obesity person.
Wistar rat: the most common with Wistar rat in the MS animal model of diet induced, conventional method has the nursing of high fat height salt feeding, high Fructose-Fed, high fat, the high sugar of high fat is fed and high-sucrose is fed.Rat there will be obesity, with insulin resistant, hypertension, hypertriglyceridemia etc., like mankind MS feature.
Remedies for diabetes conventional clinically:
(1) Regular Insulin and analogue thereof
Insulin preparation experienced by the Regular Insulin of animal-origin and genetically engineered insulin human's stage, the exploitation of the insulin analog be developed so far its using method and corresponding apparatus in addition, the physiologic secretion making ectogenic Regular Insulin also can reach simulation normal people endogenous insulin produces corresponding physiological regulation function with the fluctuation situation of blood sugar.The excellent of such as Li Lai company is secreted happy (Lispro, Humalog).
(2) insulinotropic hormone excretion
Sulfonylurea medicine: all contain a sulfonylurea group in sulfonylureas (SUD) chemical structure, its action character is sulfonylurea acceptor (SUR) specific binding with human pancreatic islet B-cell, close cytolemma potassium-channel, cell membrane potential is caused to change, open calcium channel, make flow of calcium ions, impel the secretion of Regular Insulin to increase.SUD comprises the D-860 and chlorine sulphur phenylurea etc. of the first-generation, s-generation glyburide, gliclazide, gliquidone, Glipizide etc., the glimepiride etc. of the third generation.
Non-sulfonylurea medicine: non-sulfonylurea pancreotropic hormone antidiabetic drug not only has amino acid in the structure, and show on target site, promote insulin secretion rapidly with " open soon-speed close ", reduce glycolated hemoglobin (HbA1c) and the postprandial blood sugar (PBG) of patients with NIDDM.N1,N1-Dimethylbiguanide, the thiazolidinediones medicine combined utilization of itself and binding mode complementation, so as to alleviating islet B-cells load, postpone the existence of islet cells, remarkable curative effect is shown to isolatism postprandial hyperglycemia person (IPH), First-phase of insulin secretion obstacle person and the irregular person of diet.Be described as " blood sugar regulator used during user having meals ".Medicine conventional clinically has repaglinide, nateglinide and mitiglinide etc.
(3) reduce or delay the medicine of carbohydrate metabolism and absorption
Alpha-glucosidase inhibitor: by suppressing the activity of the small intestinal mucosa chorion alpha-glucosidase of small intestine epimere, block carbohydrate breakdown and become glucose, the carbohydrate be not decomposed is hydrolyzed into glucose to the latter half of of small intestine.Thus effectively control postprandial blood sugar, remain on a more stable level.This type of medicine has acarbose, miglitol, voglibose etc.
Amylin: natural human amylin (amylin, AC-0137), for having 37 amino acid whose polypeptide hormones, is discharged by islet B-cells, effectively can suppress gastric emptying together with Regular Insulin, but unstable.Easily be hydrolyzed in vivo or aggegation.Amylin company develops its analogue pramlintide acetate.
(4) euglycemic agent
Thiazolidinediones: target spot is mainly fatty tissue, skeletal muscle and liver; energy inducing adipocyte is divided into the little adipocyte to insulin sensitivity; and the genetic transcriptions such as lipoprotein lipase are regulated on the hereditary passage of fat; to cardiac muscle and endotheliocyte, the vascular smooth muscle cell of blood vessel, to the stimulation of vasodilator substance, suppress the hyperplasia of picked-up to calcium and unstriated muscle; protection vascular system; reduce blood pressure, the damage of anti-hemostatic tube and atherosclerosis etc., have effect.This type of medicine has rosiglitazone, pioglitazone etc.
Nonthiazolidinedione class: be economic benefits and social benefits PPAR agonist, have BMS-298585, JTT-501, LY-818, DRF-2725, NN-622 etc., this type of medicine is in clinical each conceptual phase, not yet goes on the market.
(5) newly-developed hypoglycemic agent
Antidiabetic medicine novel both at home and abroad roughly comprises GLP-1 analogue, DPP-IV inhibitor, SGLT-2 inhibitor, 11 beta-HSD 1 inhibitors, G-protein linked receptor and glucokinase inhibitors etc. at present.
Berberine (berberine) has resisting pathogenic microbes effect: anti-microbial effect, antivirus action, protozoacide and antitoxic action.Effect to cardiovascular systems: anti-arrhythmia, reduce blood pressure, positive inotropic and have provide protection to ischemic brain injury.Hypoglycemic activity: a large amount of pharmacology and clinical research confirmation, Berberine not only has significant hypoglycemic activity, and to diabetics with complication hypertension thrombosis etc. have good preventive and therapeutic effect.Anti-inflammatory action, anticoagulant effect, strengthens immunologic function, anticancer.Other effects: antiulcer action, refrigeration function and also there is the effect such as CNS inhibition and cholagogic.
It is of flaccid muscles that magnolol (Magnolol) has obvious, lasting central, Central nervous depressant, and anti-inflammatory is antibacterial, resisting pathogenic microbes, and antiulcer agent is anti-oxidant, antitumor, suppresses morphine withdrawal syndrome, can the pharmacological action such as anticoagulant.Be used for the treatment of acute enteritis, bacillary or amebic dysentery.Chronic gastritis etc.Wherein, in anti-microbial effect, magnolol has significant anti-microbial activity to gram-positive bacteria, acid resistance bacterium, filamentous fungus, has more significant anti-microbial effect to streptococcus mutans, the strongest to staphylococcic restraining effect.Mainly clinically be used as to eliminate the medicines such as full vexed, the calm nervus centralis of chest abdomen, sportsmen are of flaccid muscles, antimycotic, antiulcer agent.
The coptis and the bark of official magnolia form a Chinese medicines pair; ancient prescription has " coptis bark of official magnolia soup " for representing the prescription of multiple coptis magnolia obovata composition, and the representative composition of the coptis is Berberine (berberine), and the representative composition of the bark of official magnolia is magnolol; there is antisepsis and anti-inflammation; anti-oxidant, CNS inhibition, anticancer; anticoagulant and ischemic brain injury have provide protection; relaxing smooth muscle and hypotensive effect, common or synergy, can heighten the effect of a treatment.
" natural inner core molecule " and " new multinuclear molecule " is defined as follows:
Natural inner core molecule: derive from nature biotechnology evolutionary process in order to defend certain injury or obtain certain interests through evolution, be derived (no matter its molecular size range, how complicated structure is, include primary, secondary metabolite) the molecule having central role, its core object that is derivative, that evolve is for certain injury of defence or obtains certain interests.
New multinuclear molecule: the recruit of the artificial multiple-effect through manually two or more " natural inner core molecule " synthesis or splicing being obtained.
Based on the above-mentioned definition to " natural inner core molecule ", then be there is through synthetic or " the new multinuclear molecule " that be spliced by plural " natural inner core molecule " characteristic of single " natural inner core molecule ", have the pharmacodynamic characteristics of multiple " natural inner core molecule " simultaneously concurrently; For " natural inner core molecule ", the effect of synergy, attenuation is at least served through synthetic or spliced " new multinuclear molecule ".Any key or assignment key of mononuclear molecule connect and acts on identical or diverse monokaryon (or multinuclear) molecule its molecular structure, sterie configuration are changed; Effect core also changes.
Chinese medicines is instruct the molecule how making two and plural " natural inner core molecule " it be interconnected to have multiple central role through synthetic or splicing (may be any key or assignment key) to the core concept that splicing is theoretical, is called " new multinuclear molecule ".It is different from " pharmacophore " and " twin medicine " concept of chemical drug, and " pharmacophore " refers to the steric form of the constitutional features played an important role for activity; " twin medicine " refers to and two identical or different lead compounds or medicine is connected through covalent linkage, the recruit puted together, metabolism generates above two kinds of medicines and produces synergy in vivo, enhanced activity or produce new pharmacologically active, or the selectivity of raising effect.
Chinese medicines is the important research content of traditional Chinese medicine and pharmacy to compatibility, Tang's Shennong's Herbal is discussed two medicine compatibilities from different perspectives, and it is summarized in " seven human emotions and conjunction ": namely say medicine " have single passerby, have mutual reinforcement between person, have phase envoy; have mutual restraint between two drugs person; have mutual inhibition person, have opposite, have phase senilicide; these seven human emotions all, close and look it; When good with mutual reinforcement between, phase envoy, do not use the mutual inhibition, opposite; If should make poisonous, available mutual restraint between two drugs, phase senilicide, you, do not share yet." in the compatibility research of Chinese medicine, often coming across 5 utilizations mutually together of two taste medicines in prescription, is exactly medicine pair, also known as to medicine.That is two a kind of relatively more fixing collocation between taste medicine, use and often apply in pairs, its Main Function mainly heightens the effect of a treatment, weaken toxicity and side effect.By the Experiment of Compatibility research that medicine is right, the compatibility effect between medicine pair can be disclosed, explore the rules for compatibility of its attenuation synergistic, the compatibility relationships such as the mutual reinforcement between of the in-depth Chinese medicine seven human emotions, the mutual-assistance.Medicine, to being traditional Chinese medicines pair, can study the Compatibility Law between two herbal medicines, may also be " medicine to " compatibility effect mutually in the same medicine of research between different chemical composition.
The medicine of Chinese Traditional Medicine can instruct the compatibility between Chinese medicine (medicine materical crude slice) to apply to theory; The compatibility between effective ingredient in Chinese can be instructed to apply; Also the compatibility between Effective Component of Chinese Medicine can be instructed to apply; We are developed extension and are applied to interaction between research, design and guidance drug molecule " natural inner core molecule " and impact, make the compatibility interaction type of the complicated medicine of Study of Traditional Chinese Medicine medicine centering be converted into the interaction relationship of intermolecular and " the new multinuclear molecule " aspect of drugs.
At present, the patent and correlative study that become new multinuclear molecule are spliced in the chemosynthesis about Berberine and magnolol, yet there are no open or relevant report.
Summary of the invention
The object of the present invention is to provide a kind of completely newly synthesize by berberrubine and magnolol the multinuclear molecular compound obtained; Another object of the present invention is to the preparation method that a kind of described multinuclear molecular compound is provided; Another object of the present invention is to provide the purposes of a kind of described multinuclear molecular compound in preparation treatment diabetes medicament.
For achieving the above object, the technical scheme that the present invention takes is: a kind of multinuclear molecular compound, and described multinuclear molecular compound is synthesized by berberrubine and magnolol.
Present inventor proposes the concept of multinuclear molecule first, and propose berberrubine and magnolol to synthesize or splice first to obtain a kind of brand-new multinuclear molecular compound, through checking, the toxicity of the multinuclear molecular compound obtained significantly reduces, and have significant blood sugar reducing function to Diabetes Gene mouse model, may be used for the glucose-lowering treatment of diabetes, there is good effect.
As the preferred implementation of multinuclear molecular compound of the present invention, the structural formula of described multinuclear molecular compound is:
Described n is the integer of 1≤n≤30.
As the preferred implementation of multinuclear molecular compound of the present invention, in the structural formula of multinuclear molecular compound described above, n is 2.
As the preferred implementation of multinuclear molecular compound of the present invention, described multinuclear molecular compound is prepared from by following methods:
(1a) synthesis of berberrubine derivative: take berberrubine, add organic solvent, adds X (CH after being heated to boiling 2) ny, back flow reaction, concentrates reaction solution, crystallisation by cooling, filters, obtains berberrubine derivative;
(2a) get the berberrubine derivative that step (1a) obtains to mix with magnolol, add anhydrous sodium carbonate and organic solvent, stir, heating reflux reaction, filters reacting liquor while hot, filtrate recycling design, dissolve with DMSO, post is separated, and uses the methanol solution wash-out of 30%, 40%, 50%, 60% successively, HPLC detects, and collects 60% methanol-eluted fractions, concentrated, sample mixed by concentrated solution recycle silicon glue, silicagel column is separated, and wash-out, obtains multinuclear molecular compound;
X (CH described in described step (1a) 2) nin Y, X=Y or X ≒ Y, X, Y are O, S, F, Cl, Br or I, and n is the integer of 1≤n≤30.
Preferably, the organic solvent adopted in described step (1a) is acetonitrile, and certain those skilled in the art also can select other suitable materials as organic solvent according to prior art.Synthesized by berberrubine in described step (1a) that to obtain the reaction process of berberrubine derivative as follows:
In above-mentioned reaction formula, n is the integer of 1≤n≤30.
Preferably, to react by berberrubine derivative and magnolol the reaction process generating multinuclear molecular compound as follows for described step (2a):
Wherein, n is the integer of 1≤n≤30.
As the preferred implementation of multinuclear molecular compound of the present invention, the building-up process of the red alkali derivant of described step (1a) Berberine is: take berberrubine 1 weight part, add organic solvent 150 ~ 200 weight part, 85 DEG C are heated to boiling, add 1, 2-ethylene dibromide 20 ~ 40 weight part, back flow reaction 3h, reaction solution is concentrated into 50 ~ 100 parts, crystallisation by cooling, filter, with appropriate organic solvent washing crystallization, washings and filtrate merge, reclaim organic solvent, the residue dissolve with methanol of 30 weight parts, crystallisation by cooling, filter, use methanol wash crystallization, merge with above-mentioned crystallization, obtain berberrubine derivative.As described X (CH 2) nwhen Y is glycol dibromide, it is berberrubine-9-oxygen monobromoethane that berberrubine and glycol dibromide react the berberrubine derivative generated, and its chemical structural formula is as follows:
As multinuclear molecular compound preferred implementation of the present invention, described step (2a) is: get berberrubine derivative 1.2 weight part that step (1a) obtains and mix with magnolol 1 weight part, add anhydrous sodium carbonate 2 weight part, organic solvent 150 weight part, stir, be heated to 85 DEG C, backflow 8h, reacting liquor while hot is filtered, filtrate recycling design, with the DMSO solvent of 5 weight parts, post is separated, use 30% successively, 40%, 50%, the methanol solution wash-out of 60%, HPLC detects, collect 60% methanol-eluted fractions, concentrated, the silica gel mixed sample of 3 weight parts used again by concentrated solution, silicagel column is separated, with sherwood oil and the ethyl acetate mixtures wash-out of 20 times of column volumes, use methanol-eluted fractions again, reclaim methyl alcohol, obtain multinuclear molecular compound.Preferably, C18 post is adopted to be separated; The granular size of described silica gel is 400-500 order; In described sherwood oil and ethyl acetate mixtures, the volume ratio of sherwood oil and ethyl acetate is 1:1.The multinuclear molecular compound obtained, it is 615.3 that MS obtains molecular weight, and fusing point is 145.2 ~ 146.1.
As the another preferred implementation of multinuclear molecular compound of the present invention, described multinuclear molecular compound is prepared from by following methods:
(1b) synthesis of magnolia bark phenol derivative: get magnolol, mix with anhydrous sodium carbonate, add organic solvent, then add X (CH 2) ny, reacting by heating, obtains magnolia bark phenol derivative;
(2b) get magnolia bark phenol derivative that step (1b) obtains, anhydrous sodium carbonate, berberrubine mixing, add organic solvent, stir, heating reflux reaction, filters reacting liquor while hot, filtrate recycling design, dissolve with DMSO, post is separated, and uses the methanol solution wash-out of 30%, 40%, 50%, 60% successively, HPLC detects, and collects 60% methanol-eluted fractions, concentrated, sample mixed by concentrated solution recycle silicon glue, silicagel column is separated, and wash-out, obtains multinuclear molecular compound;
X (CH described in described step (1b) 2) nin Y, X=Y or X ≒ Y, X, Y are O, S, F, Cl, Br or I, and n is the integer of 1≤n≤30.
As follows by the reaction process of magnolol synthesis magnolia bark phenol derivative in described step (1b):
In above-mentioned reaction formula, n is the integer of 1≤n≤30.
The reaction process that described step (2b) synthesizes multinuclear molecular compound by berberrubine and magnolia bark phenol derivative is as follows:
Wherein, n is the integer of 1≤n≤30.
As the preferred implementation of multinuclear molecular compound of the present invention, magnolol, anhydrous sodium carbonate, X (CH in described step (1b) 2) nthe mol ratio of Y is 1:2:16, and described magnolol is 1/150mol/L with the molecular volume ratio of organic solvent, and temperature of reaction is 85 DEG C, and the reaction times is 5h; Described X (CH 2) ny is glycol dibromide.As described X (CH 2) nwhen Y is glycol dibromide, the magnolia bark phenol derivative generated is magnolol-1-oxygen monobromoethane, and its structural formula is as follows:
As the preferred implementation of multinuclear molecular compound of the present invention, described step (2b) is: get magnolia bark phenol derivative 1 weight part that step (1b) obtains, anhydrous sodium carbonate 2 weight part, berberrubine 1 weight part adds in reactor, add organic solvent 150 parts, stir, be heated to 85 DEG C, backflow 8h, reacting liquor while hot is filtered, filtrate recycling design, with the DMSO solvent of 5 weight parts, post is separated, use 30% successively, 40%, 50%, the methanol solution wash-out of 60%, HPLC detects, collect 60% methanol-eluted fractions, concentrated, the silica gel mixed sample of 3 weight parts used again by concentrated solution, silicagel column is separated, with sherwood oil and the ethyl acetate mixtures wash-out of 20 times of column volumes, use methanol-eluted fractions again, reclaim methyl alcohol, obtain multinuclear molecular compound.Preferably, the granular size of described silica gel is 400-500 order; In described sherwood oil and ethyl acetate mixtures, the volume ratio of sherwood oil and ethyl acetate is 1:1.
Organic solvent in step described above (1a), (2a), (1b), (2b), those skilled in the art can carry out suitable selection according to prior art, and preferably, described organic solvent is acetonitrile.
Berberrubine described above can be berberrubine of the prior art, and those skilled in the art can take suitable approach to obtain, such as, synthesize or be directly purchased from market.Preferably, the berberrubine in the present invention adopts following methods to be prepared from: add Berberine and DMF that solid-liquid ratio is 1:15 ~ 30g/L in the reactor, add zeolite, reflux condensation mode, under 400W ~ 800W microwave radiation, reaction 10 ~ 20min, takes out reactor, add water dilution cooling while hot, refrigerated overnight, makes crystallization complete, suction filtration, drying, obtains crystallization a; Filtrate is separately separated by macroporous resin column, uses the methanol-eluted fractions of 40%, 45%, 50%, 55%, 60%, 65% and 70% successively, collects 70% methanol-eluted fractions position, concentrated, obtains crystallization b, is merged by crystallization a and crystallization b, obtain berberrubine.The chemical structural formula of described berberrubine is as follows:
Another object of the present invention is to the preparation method providing a kind of multinuclear molecular compound described above, for realizing this object, the technical scheme that the present invention takes is: a kind of preparation method of multinuclear molecular compound, comprises the following steps:
(1a) synthesis of berberrubine derivative: take berberrubine, add organic solvent, adds X (CH after being heated to boiling 2) ny, back flow reaction, concentrates reaction solution, crystallisation by cooling, filters, obtains berberrubine derivative;
(2a) get the berberrubine derivative that step (1a) obtains to mix with magnolol, add anhydrous sodium carbonate and organic solvent, stir, heating reflux reaction, filters reacting liquor while hot, filtrate recycling design, dissolve with DMSO, post is separated, and uses the methanol solution wash-out of 30%, 40%, 50%, 60% successively, HPLC detects, and collects 60% methanol-eluted fractions, concentrated, sample mixed by concentrated solution recycle silicon glue, silicagel column is separated, and wash-out, obtains multinuclear molecular compound;
X (CH described in described step (1a) 2) nin Y, X=Y or X ≒ Y, X, Y are O, S, F, Cl, Br or I, and n is the integer of 1≤n≤30.
As the preferred implementation of the preparation method of the present invention's multinuclear molecular compound described above, the building-up process of the red alkali derivant of described step (1a) Berberine is: take berberrubine 1 weight part, add organic solvent 150 ~ 200 weight part, 85 DEG C are heated to boiling, add 1, 2-ethylene dibromide 20 ~ 40 weight part, back flow reaction 3h, reaction solution is concentrated into 50 ~ 100 parts, crystallisation by cooling, filter, with appropriate organic solvent washing crystallization, washings and filtrate merge, reclaim organic solvent, the residue dissolve with methanol of 30 weight parts, crystallisation by cooling, filter, use methanol wash crystallization, merge with above-mentioned crystallization, obtain berberrubine derivative.
As the preferred implementation of the preparation method of the present invention's multinuclear molecular compound described above, described step (2a) is: get berberrubine derivative 1.2 weight part that step (1a) obtains and mix with magnolol 1 weight part, add anhydrous sodium carbonate 2 weight part, organic solvent 150 weight part, stir, be heated to 85 DEG C, backflow 8h, reacting liquor while hot is filtered, filtrate recycling design, with the DMSO solvent of 5 weight parts, post is separated, use 30% successively, 40%, 50%, the methanol solution wash-out of 60%, HPLC detects, collect 60% methanol-eluted fractions, concentrated, the silica gel mixed sample of 3 weight parts used again by concentrated solution, silicagel column is separated, with sherwood oil and the ethyl acetate mixtures wash-out of 20 times of column volumes, use methanol-eluted fractions again, reclaim methyl alcohol, obtain multinuclear molecular compound.
Another object of the present invention is the preparation method providing another kind of multinuclear molecular compound as mentioned above, and for realizing this object, the technical scheme that the present invention takes is: a kind of preparation method of multinuclear molecular compound, comprises the following steps:
(1b) synthesis of magnolia bark phenol derivative: get magnolol, mix with anhydrous sodium carbonate, add organic solvent, then add X (CH 2) ny, reacting by heating, obtains magnolia bark phenol derivative;
(2b) get magnolia bark phenol derivative that step (1b) obtains, anhydrous sodium carbonate, berberrubine mixing, add organic solvent, stir, heating reflux reaction, filters reacting liquor while hot, filtrate recycling design, dissolve with DMSO, post is separated, and uses the methanol solution wash-out of 30%, 40%, 50%, 60% successively, HPLC detects, and collects 60% methanol-eluted fractions, concentrated, sample mixed by concentrated solution recycle silicon glue, silicagel column is separated, and wash-out, obtains multinuclear molecular compound;
X (CH described in described step (1b) 2) nin Y, X=Y or X ≒ Y, X, Y are O, S, F, Cl, Br or I, and n is the integer of 1≤n≤30.
As the preferred implementation of the another kind of preparation method of multinuclear molecular compound of the present invention, magnolol, anhydrous sodium carbonate, X (CH in described step (1b) 2) nthe mol ratio of Y is 1:2:8, and described magnolol is 1/150mol/L with the molecular volume ratio of organic solvent, and temperature of reaction is 85 DEG C, and the reaction times is 5h; Described X (CH 2) ny is glycol dibromide.
As the preferred implementation of the another kind of preparation method of multinuclear molecular compound of the present invention, described step (2b) is: get magnolia bark phenol derivative 1 weight part that step (1b) obtains, anhydrous sodium carbonate 2 weight part, berberrubine 1 weight part adds in reactor, add organic solvent 150 parts, stir, be heated to 85 DEG C, backflow 8h, reacting liquor while hot is filtered, filtrate recycling design, with the DMSO solvent of 5 weight parts, post is separated, use 30% successively, 40%, 50%, the methanol solution wash-out of 60%, HPLC detects, collect 60% methanol-eluted fractions, concentrated, the silica gel mixed sample of 3 weight parts used again by concentrated solution, silicagel column is separated, with sherwood oil and the ethyl acetate mixtures wash-out of 20 times of column volumes, use methanol-eluted fractions again, reclaim methyl alcohol, obtain multinuclear molecular compound.
Another object of the present invention is the purposes providing a kind of multinuclear molecular compound described above in the medicine of preparation treatment diabetes.
The invention provides a kind of brand-new material multinuclear molecular compound, described multinuclear molecular compound is also the concept that present inventor proposes first, this multinuclear molecular compound is synthesized by berberrubine and magnolol and obtains, change the absorption of single component or two constituents mixts, distribution, the drug metabolism link of metabolism and excretion, gained to multinuclear molecular compound compared with Berberine, toxicity only has 1/10th of Berberine, toxicity is significantly reduced, absorption significantly improves, improve bioavailability, extend the metabolism time, long-acting effect can be reached, and hypoglycemic effect obtains enhancing.The preparation method of described multinuclear molecular compound provided by the invention, technique is simple, and easy handling, is convenient to large-scale industrial production.The purposes of described multinuclear molecular compound provided by the invention in preparation treatment diabetes medicament, have significant hypoglycemic effect, the clinical treatment for diabetes provides how efficient, safe drug candidate, to meet many-sided demand of clinical treatment.
Accompanying drawing explanation
Fig. 1 is the chemical structure schematic diagram of multinuclear molecular compound of the present invention.
Fig. 2 is berberrubine 1h NMR schemes;
Fig. 3 is berberrubine 13c NMR schemes;
Fig. 4 is berberrubine-9-oxygen monobromoethane 1h NMR schemes;
Fig. 5 is berberrubine-9-oxygen monobromoethane 13c NMR schemes;
Fig. 6 is multinuclear molecular compound of the present invention 1h NMR schemes;
Fig. 7 is multinuclear molecular compound of the present invention 13c NMR schemes;
Fig. 8 is aP2-SREBP-1c murine genes type qualification figure;
Fig. 9 is that multinuclear molecular compound of the present invention is on the impact of aP2-SREBP-1c mouse blood sugar;
Figure 10 is that multinuclear molecular compound of the present invention is on the impact of aP2-SREBP-1c mouse AUC;
Figure 11 is that multinuclear molecular compound of the present invention is on the impact of aP2-SREBP-1c mouse TG;
Figure 12 is the impact of multinuclear molecular compound of the present invention on the type II diabetes mouse blood sugar that STZ+ height fat causes;
Figure 13 is the impact of multinuclear molecular compound of the present invention on the type II diabetes mouse AUC that STZ+ height fat causes;
Figure 14 is the impact of multinuclear molecular compound of the present invention on the type II diabetes mouse TG that STZ+ height fat causes.
Embodiment
For better the object, technical solutions and advantages of the present invention being described, below in conjunction with the drawings and specific embodiments, the invention will be further described.
Embodiment 1
A kind of embodiment of multinuclear molecular compound of the present invention, multinuclear molecular compound described in the present embodiment adopts following methods to be prepared from:
(1a) synthesis of berberrubine derivative: take berberrubine 1 weight part, adds acetonitrile 150 weight part, and 85 DEG C are heated to boiling, add X (CH 2) ny40 weight part, back flow reaction 3h, is concentrated into 50 parts by reaction solution, crystallisation by cooling, filters, with appropriate acetonitrile wash crystallization, washings and filtrate merge, and reclaim acetonitrile, the residue dissolve with methanol of 30 weight parts, crystallisation by cooling, filter, use methanol wash crystallization, merge with above-mentioned crystallization, obtain that sample yield is 56.34%, the berberrubine derivative of purity more than 95%, wherein said X (CH 2) nin Y, X, Y are Br, n=2, and gained berberrubine derivative is berberrubine-9-oxygen monobromoethane, its 1h NMR scheme and 13c NMR schemes respectively as shown in figs. 4 and 5, and wherein in Fig. 4, the ownership of H is in table 1:
Table 1
In Fig. 5, the ownership of C is in table 2:
Table 2
(2a) get berberrubine derivative 1.2 weight part that step (1a) obtains to mix with magnolol 1 weight part, add anhydrous sodium carbonate 2 weight part, acetonitrile 150 weight part, stir, be heated to 85 DEG C, backflow 8h, reacting liquor while hot is filtered, filtrate recycling design, with the DMSO solvent of 5 weight parts, post is separated, use 30% successively, 40%, 50%, the methanol solution wash-out of 60%, HPLC detects, collect 60% methanol-eluted fractions, concentrated, concentrated solution uses the silica gel of 3 weight parts (400-500 order) to mix sample again, silicagel column is separated, with sherwood oil and ethyl acetate mixtures (volume ratio of sherwood oil and ethyl acetate the is 1:1) wash-out of 20 times of column volumes, use methanol-eluted fractions again, reclaim methyl alcohol, obtain multinuclear molecular compound, the yield of described multinuclear molecule is 22 ~ 28%, it is 615.3 that MS obtains molecular weight, fusing point is 145.2 ~ 146.1, the molecular structural formula of gained multinuclear molecular compound as shown in Figure 1, gained multinuclear molecular compound 1h NMR scheme and 13c NMR schemes respectively as seen in figs. 6 and 7, and wherein in accompanying drawing 6, the ownership of H is as table 3:
In accompanying drawing 7, the ownership of C is in table 4:
Table 4
Berberrubine in step described in the present embodiment (1a) can adopt either method in prior art to obtain, also can directly from market purchase etc., preferably, berberrubine in step described in the present embodiment (1a) adopts following methods to be prepared from: add Berberine and DMF that solid-liquid ratio is 1:15 ~ 30g/L in the reactor, add zeolite, reflux condensation mode, under 400W ~ 800W microwave radiation, reaction 10 ~ 20min, takes out reactor, add water dilution cooling while hot, refrigerated overnight, makes crystallization complete, suction filtration, drying, obtains crystallization a; Filtrate is separately separated by macroporous resin column, use the methanol-eluted fractions of 40%, 45%, 50%, 55%, 60%, 65% and 70% successively, collect 70% methanol-eluted fractions position, concentrated, obtain crystallization b, crystallization a and crystallization b is merged, obtain berberrubine, total yield of products is 80 ~ 95%, purity>=98% of gained berberrubine, it is 321.2 that MS obtains molecular weight, its 1h NMR scheme and 13c NMR schemes respectively as shown in Figures 2 and 3, and wherein in accompanying drawing 2, the ownership of H is in table 5:
Table 5
In accompanying drawing 3, the ownership of C is in table 6:
Table 6
Embodiment 2
A kind of embodiment of multinuclear molecular compound of the present invention, multinuclear molecular compound described in the present embodiment adopts following methods to be prepared from:
(1a) synthesis of berberrubine derivative: take berberrubine 1 weight part, adds acetonitrile 200 weight part, and 85 DEG C are heated to boiling, add X (CH 2) ny20 weight part, back flow reaction 3h, is concentrated into 100 parts by reaction solution, crystallisation by cooling, filters, with appropriate acetonitrile wash crystallization, washings and filtrate merge, and reclaim acetonitrile, the residue dissolve with methanol of 30 weight parts, crystallisation by cooling, filter, use methanol wash crystallization, merge with above-mentioned crystallization, obtain that sample yield is 56.34%, the berberrubine derivative of purity more than 95%, wherein said X (CH 2) nin Y, X, Y are Cl, n=3;
(2a) get berberrubine derivative 1.2 weight part that step (1a) obtains to mix with magnolol 1 weight part, add anhydrous sodium carbonate 2 weight part, acetonitrile 150 weight part, stir, be heated to 85 DEG C, backflow 8h, reacting liquor while hot is filtered, filtrate recycling design, with the DMSO solvent of 5 weight parts, post is separated, use 30% successively, 40%, 50%, the methanol solution wash-out of 60%, HPLC detects, collect 60% methanol-eluted fractions, concentrated, concentrated solution uses the silica gel of 3 weight parts (400-500 order) to mix sample again, silicagel column is separated, with sherwood oil and ethyl acetate mixtures (volume ratio of sherwood oil and ethyl acetate the is 1:1) wash-out of 20 times of column volumes, use methanol-eluted fractions again, reclaim methyl alcohol, obtain multinuclear molecular compound, the yield of described multinuclear molecule is 22 ~ 28%, it is 615.3 that MS obtains molecular weight, fusing point is 145.2 ~ 146.1, the molecular structural formula of gained multinuclear molecular compound as shown in Figure 1.
Berberrubine in step described in the present embodiment (1a) can adopt either method in prior art to obtain, also can directly from market purchase etc., preferably, the berberrubine in step described in the present embodiment (1a) adopts the method described in embodiment 1 to be prepared from.
Embodiment 3
A kind of embodiment of multinuclear molecular compound of the present invention, multinuclear molecular compound described in the present embodiment adopts following methods to be prepared from:
(1a) synthesis of berberrubine derivative: take berberrubine 1 weight part, adds acetonitrile 160 weight part, and 85 DEG C are heated to boiling, add X (CH 2) ny30 weight part, back flow reaction 3h, is concentrated into 80 parts by reaction solution, crystallisation by cooling, filters, with appropriate acetonitrile wash crystallization, washings and filtrate merge, and reclaim acetonitrile, the residue dissolve with methanol of 30 weight parts, crystallisation by cooling, filter, use methanol wash crystallization, merge with above-mentioned crystallization, obtain that sample yield is 56.34%, the berberrubine derivative of purity more than 95%, wherein said X (CH 2) nin Y, X, Y are S, n=4;
(2a) get berberrubine derivative 1.2 weight part that step (1a) obtains to mix with magnolol 1 weight part, add anhydrous sodium carbonate 2 weight part, acetonitrile 150 weight part, stir, be heated to 85 DEG C, backflow 8h, reacting liquor while hot is filtered, filtrate recycling design, with the DMSO solvent of 5 weight parts, post is separated, use 30% successively, 40%, 50%, the methanol solution wash-out of 60%, HPLC detects, collect 60% methanol-eluted fractions, concentrated, concentrated solution uses the silica gel of 3 weight parts (400-500 order) to mix sample again, silicagel column is separated, with sherwood oil and ethyl acetate mixtures (volume ratio of sherwood oil and ethyl acetate the is 1:1) wash-out of 20 times of column volumes, use methanol-eluted fractions again, reclaim methyl alcohol, obtain multinuclear molecular compound, the yield of described multinuclear molecule is 22 ~ 28%, it is 615.3 that MS obtains molecular weight, fusing point is 145.2 ~ 146.1, the molecular structural formula of gained multinuclear molecular compound as shown in Figure 1.
Berberrubine in step described in the present embodiment (1a) can adopt either method in prior art to obtain, also can directly from market purchase etc., preferably, the berberrubine in step described in the present embodiment (1a) adopts the method described in embodiment 1 to be prepared from.
Embodiment 4
A kind of embodiment of multinuclear molecular compound of the present invention, multinuclear molecular compound described in the present embodiment adopts following methods to be prepared from:
(1a) synthesis of berberrubine derivative: take berberrubine 1 weight part, adds acetonitrile 180 weight part, and 85 DEG C are heated to boiling, add X (CH 2) ny35 weight part, back flow reaction 3h, is concentrated into 60 parts by reaction solution, crystallisation by cooling, filters, with appropriate acetonitrile wash crystallization, washings and filtrate merge, and reclaim acetonitrile, the residue dissolve with methanol of 30 weight parts, crystallisation by cooling, filter, use methanol wash crystallization, merge with above-mentioned crystallization, obtain that sample yield is 56.34%, the berberrubine derivative of purity more than 95%, wherein said X (CH 2) nin Y, X, Y are O, n=1;
(2a) get berberrubine derivative 1.2 weight part that step (1a) obtains to mix with magnolol 1 weight part, add anhydrous sodium carbonate 2 weight part, acetonitrile 150 weight part, stir, be heated to 85 DEG C, backflow 8h, reacting liquor while hot is filtered, filtrate recycling design, with the DMSO solvent of 5 weight parts, post is separated, use 30% successively, 40%, 50%, the methanol solution wash-out of 60%, HPLC detects, collect 60% methanol-eluted fractions, concentrated, concentrated solution uses the silica gel of 3 weight parts (400-500 order) to mix sample again, silicagel column is separated, with sherwood oil and ethyl acetate mixtures (volume ratio of sherwood oil and ethyl acetate the is 1:1) wash-out of 20 times of column volumes, use methanol-eluted fractions again, reclaim methyl alcohol, obtain multinuclear molecular compound, the yield of described multinuclear molecule is 22 ~ 28%, it is 615.3 that MS obtains molecular weight, fusing point is 145.2 ~ 146.1, the molecular structural formula of gained multinuclear molecular compound as shown in Figure 1.
Berberrubine in step described in the present embodiment (1a) can adopt either method in prior art to obtain, also can directly from market purchase etc., preferably, the berberrubine in step described in the present embodiment (1a) adopts the method described in embodiment 1 to be prepared from.
Embodiment 5
A kind of embodiment of multinuclear molecular compound of the present invention, multinuclear molecular compound described in the present embodiment adopts following methods to be prepared from:
(1b) synthesis of magnolia bark phenol derivative: get magnolol 1 molar part, mixes with anhydrous sodium carbonate 2 molar part, adds acetonitrile, and described acetonitrile is 150:1(L/mol with the Molar ratio of magnolol), then add X (CH 2) ny16 molar part, 85 DEG C of reaction 5h obtain that yield is 28%, purity is the magnolia bark phenol derivative of 95%, wherein said X (CH 2) nin Y, X, Y are I, n=10;
(2b) magnolia bark phenol derivative 1 weight part that step (1b) obtains is got, anhydrous sodium carbonate 2 weight part, berberrubine 1 weight part adds in reactor, add acetonitrile 150 parts, stir, be heated to 85 DEG C, backflow 8h, reacting liquor while hot is filtered, filtrate recycling design, with the DMSO solvent of 5 weight parts, post is separated, use 30% successively, 40%, 50%, the methanol solution wash-out of 60%, HPLC detects, collect 60% methanol-eluted fractions, concentrated, concentrated solution uses the silica gel of 3 weight parts (400-500 order) to mix sample again, silicagel column is separated, with sherwood oil and ethyl acetate mixtures (volume ratio of sherwood oil and ethyl acetate the is 1:1) wash-out of 20 times of column volumes, use methanol-eluted fractions again, reclaim methyl alcohol, obtain multinuclear molecular compound, the yield of described multinuclear molecule is 22 ~ 28%, it is 615.3 that MS obtains molecular weight, fusing point is 145.2 ~ 146.1, the molecular structural formula of gained multinuclear molecular compound as shown in Figure 1.
Berberrubine in step described in the present embodiment (2b) can adopt either method in prior art to obtain, also can directly from market purchase etc., preferably, the berberrubine in step described in the present embodiment (2b) adopts the method described in embodiment 1 to be prepared from.
Embodiment 6
A kind of embodiment of multinuclear molecular compound of the present invention, multinuclear molecular compound described in the present embodiment adopts the method identical with embodiment 5 to be prepared from, except described X (CH 2) nin Y, X, Y are F, and outside n=30, all the other are all identical with embodiment 5.
Embodiment 7
A kind of embodiment of multinuclear molecular compound of the present invention, multinuclear molecular compound described in the present embodiment adopts the method identical with embodiment 5 to be prepared from, except described X (CH 2) nin Y, X, Y are Br, and outside n=18, all the other are all identical with embodiment 5.
Embodiment 8
Multinuclear molecular compound of the present invention is on the impact of aP2-SREBP-1c transgenic mice blood sugar, blood fat
1 materials and methods
1.1 animals are introduced
AP2-SREBP-1c transgenic mice is introduced by U.S. Jackson laboratory, entrusts the purification of model animal institute of Nanjing University, then goes to Traditional Chinese Medicine University Of Guangzhou's captive breeding.Conformity certification number: J003393SCXK (Soviet Union) 2010-0001.Captive breeding condition: Traditional Chinese Medicine University Of Guangzhou Experimental Animal Center SPF level Animal House, temperature 20 ~ 25 DEG C, humidity 50 ~ 80%, 12h:12h intermittent illumination round the clock, normal diet is bought in Guangdong Medical Lab Animal Center.
1.2 animal features
SREBP-1c is important a member of nuclear factor family, and it mainly participates in the expression of lipogenesis and glucose metabolism relative enzyme gene.It has three kinds of forms: SREBP-1C, SREBP-1a and SREBP-2.SREBP-2 tends to activate the synthesis of cholesterol, SREBP-1a and SREBP-1c tends to promote the synthesis of lipid acid and SREBP-1c predominant expression in liver and fatty tissue.This transgenic mouse lines utilizes adenovirus technology to make SREBP-1c overexpression in fatty tissue, it is similar to another kind of adipose tissue development obstacle transgenic models mouse A-ZIP/F-1 mouse, show obvious white adipose atrophy when the former is born, brown fat is flourishing.Latter shows as white adipose and definitely lacks, and brown fat significantly reduces, and the two all occurs that a series of metabolism syndrome disease comprises insulin resistant, hyperlipidaemia, hyperglycemia etc.
1.3 animal reproduction
More than 6 week age, agematched Male Transgenic type and female wild type mouse mate with 1:2, and after female mouse pregnancy, male mouse divides cage separately, and conceived female mouse is suitably added nutrition and feeds and raise, and rearing conditions is the same.Mouse is cut tail and is about the capable genotype identification of 1cm after being born 2 weeks, distinguish wild-type and transgenic.
1.4 genotype identification
AP2-SREBP-1c transgenic mice is the heterozygote individual of C57BL/6J and SJL background, mouse tail DNA extraction and pcr amplification is carried out with reference to Jackson laboratory genotype identification method, qualification transgenic (transgene-type, T) and wild-type (wild-type, W).
1.5 multinuclear molecular compound dosage and preparations
Dosage 40mg/kg/10ml.Take the multinuclear molecular compound powder of the arbitrary gained of 40mg embodiment 1 ~ 7, successively add 5% gum arabic solution, limit edged grinds, and final constant volume, to 10ml, to obtain final product.
1.6 animal grouping and process
Select brood female wild type mouse (W) 8 in 12 week age, transgenic mouse (T) 16.Be divided into normal group (wild-type), control group (transgenic), multinuclear molecular compound group (transgenic), often organize 8.Survey body weight and food-intake, flooding quantity each 2 times weekly, gastric infusion, control group gives isometric(al) 5% gum arabic solution, and administration volume is 10ml/kg body weight, once a day, and continuous 13 weeks.In the 8th week fasting 12h, ether was gently numb, and eyeground vein clump is got blood and surveys Glu(glucose), TG(triglyceride level), survey oral glucose tolerance (OGTT) after the 9th week fasting 12h, finally get blood by front method at the end of experiment and survey Glu, TG.
1.7 key instruments and reagent
BS110S electronic analytical balance, German Sartorius; MG96G gene-amplificative instrament, Hangzhou Lang Ji scientific instrument company limited; The long microplate reader of all-wave, Multiskan GO, 1510, Thermo Fisher Scientific Oy Ratastie2.Fi-01620Vantaa, Finland; HITACHI CR22G high speed freezing centrifuge, FDAC; D-dextrose anhydrous, BIOSHARP.Glu, TG test kit, 20130201,20130301, ShangHai RongSheng Biology Pharmacy Co., Ltd.
1.8 Testing index and method
(1) blood biochemical: the mensuration of Glu, TG is all undertaken by test kit specification sheets.(2) OGTT(oral glucose tolerance test): after animal fasting 12h, the shallow fiber crops of ether, eyeground vein clump gets blood, then gives glucose with 2g/kg body weight gavage, gets blood, survey 0,20,60,120min blood sugar.Make blood sugar-time curve, and area (AUC) under calculated curve.
1.9 statistical procedures methods
All data all with represent, every result adopts StatView software to carry out ANOVA check analysis.
2 results
2.1 genotype identification show exogenous nSREBP-1c and express
The structure of aP2-SREBP-1c transgenic mice is the DNA fragmentation of a 5.4kb of the aP2 genetic enhancer/promoters driven by inserting fatty tissue specifically expressing, this fragment coding people SREBP1c1-436 aminoacid sequence, i.e. caryogram SREBP-1c(nSREBP-1c).Genotype identification display is consistent with Jackson laboratory, and neonatal transgenic type (T) and wild-type (W) mouse ratio are about 1:1.PCR target stripe is (Transgene) 151bp, and internal reference (Internal positive control) is 324bp(Fig. 8).Shimomura etc. analyze with Northern Blot and also show the nSREBP-1c mRNA overexpression being derived from people in transgenic mouse white adipose.
2.2 multinuclear molecular compounds are on the glycometabolic impact of aP2-SREBP-1c mouse
Embodiment 1 ~ 7 gained multinuclear molecular compound all significantly can reduce 20min, 60min blood sugar and AUC in OGTT experiment, illustrates that multinuclear molecular compound tool of the present invention is significantly improved aP2-SREBP-1c mouse carbohydrate metabolism.(see Fig. 9, Figure 10)
2.3 multinuclear molecular compounds are on the lipometabolic impact of aP2-SREBP-1c mouse
Embodiment 1 ~ 7 gained multinuclear molecular compound all significantly can reduce TG in the 8th week, the 13rd week aP2-SREBP-1c Mouse Blood, illustrates that multinuclear molecular compound tool of the present invention is significantly improved the effect of aP2-SREBP-1c mouse lipid metabolism.(see Figure 11)
Embodiment 9
The type II diabetes mouse blood sugar that multinuclear molecular compound of the present invention causes STZ+ height fat, the impact of blood fat
1 materials and methods
1.1 laboratory animal C57BL/6N mouse, SPF level, is provided by Beijing Vital River Experimental Animals Technology Co., Ltd., and animal conformity certification number is SCXR(capital) 2012-0001; High lipid food is processed by Guangdong Medical Lab Animal Center, processing lot number: No.0116860, formula composition: casein 26.17%, CYSTINE 0.39%, Star Dri 5 16.35%, sucrose 9.00%, Mierocrystalline cellulose 6.54%, soya-bean oil 3.27%, lard 32.06%, mineral substance AIN-934.58%, vitamin A IN-931.31%, choline chloride 60 0.33%, cholesterol 1.0%, cholate 0.15%, feed conformity certification card number is SCXK (Guangdong) 2008-0002; Basal feed is provided by Guangdong Medical Lab Animal Center.Animal rearing is in Traditional Chinese Medicine University Of Guangzhou Experimental Animal Center SPF level Animal House, and raising temperature and humidity: 20 ~ 25 DEG C, 40 ~ 70%, adopt 12h:12h intermittent illumination round the clock, and ad lib is drunk water.
1.2 multinuclear molecular compound dosage and preparations: dosage 40mg/kg/10ml.Take the multinuclear molecular compound powder of the arbitrary gained of 40mg embodiment 1 ~ 7, successively add 5% gum arabic solution, limit edged grinds, and final constant volume, to 10ml, to obtain final product.
After 1.3 animal groupings and process C57BL/6N mouse adaptability raise 1 week, reference literature method (Kusakabe T, et al.Diabetologia, 2009,52 (4): 675-683), in fasting after 4 hours, except 10 mouse peritoneal injection solvents, the equal abdominal injection STZ120mg/kg of all the other mouse, after 3 weeks, by body weight and basic biochemistry index, mouse is divided into 3 groups at random, often organizes 10.Be respectively blank group, model group, multinuclear molecular compound group.Except blank group gives except basal feed, abdominal injection STZ mouse feeds raises 60% high lipid food (formula is with reference to Research Diet company D12492 feed).Until experiment terminates.Survey body weight and food-intake, flooding quantity each 2 times weekly, gastric infusion, control group gives isometric(al) 5% gum arabic solution, and administration volume is 10ml/kg body weight, once a day, and continuous 13 weeks.In the 8th week fasting 12h, ether was gently numb, and eyeground vein clump is got blood and surveys Glu(glucose), TG(triglyceride level), survey oral glucose tolerance (OGTT) after the 9th week fasting 12h, finally get blood by front method at the end of experiment and survey Glu, TG.
1.4 key instruments and reagent BS110S electronic analytical balance, German Sartorius; The long microplate reader of all-wave, Multiskan GO, 1510, Thermo Fisher Scientific Oy Ratastie2.Fi-01620Vantaa, Finland; HITACHI CR22G high speed freezing centrifuge, FDAC; D-dextrose anhydrous, BIOSHARP.Glu, TG test kit: 20120801,20121001, ShangHai RongSheng Biology Pharmacy Co., Ltd.Chain arteries and veins assistant rhzomorph (STZ), Sigma company, the used time dissolves with citrate buffer, by 10ml/kg abdominal injection.
1.5 Testing index and method (1) blood biochemical: the mensuration of Glu, TG is all undertaken by test kit specification sheets.(2) OGTT(oral glucose tolerance test): after animal fasting 12h, the shallow fiber crops of ether, eyeground vein clump gets blood, then gives glucose with 2g/kg body weight gavage, gets blood, survey 0,20,60,120min blood sugar.Make blood sugar-time curve, and area (AUC) under calculated curve.
1.6 statistical procedures method all datas all with represent, every result adopts StatView software to carry out ANOVA check analysis.
2. result
The glycometabolic impact of type II diabetes mouse that 2.1 multinuclear molecular compounds cause STZ+ height fat
Embodiment 1 ~ 7 gained multinuclear molecular compound all significantly can reduce 0min, 20min, 60min, 120min blood sugar and AUC in OGTT experiment, the type II diabetes mouse carbohydrate metabolism being significantly improved multinuclear molecular compound tool of the present invention STZ+ height fat causes is described, (see Figure 12, Figure 13)
2.2 multinuclear molecular compounds are on the impact of the type II diabetes lipid of mice that STZ+ height fat causes
Embodiment 1 ~ 7 gained multinuclear molecular compound all significantly can reduce TG in the 8th week, the 13rd week aP2-SREBP-1c Mouse Blood, the type II diabetes mouse lipid metabolism effect being significantly improved multinuclear molecular compound tool of the present invention STZ+ height fat causes is described, (see Figure 14).
Embodiment 10
The acute toxicity test of multinuclear molecular compound of the present invention
1 experiment material
1.1 trial-product
(1) title: the multinuclear molecular compound of the arbitrary gained of embodiment 1 ~ 7 (airtight leave in 4 DEG C of refrigerators).
(2) proterties: powder
(3) solvent used: the 0.5% tween-80 aqueous solution
(4) compound method: the emulsion being emulsified into desired concn with the 0.5% tween-80 aqueous solution.
(5) preservation condition: a time preparation is finished (4 DEG C of Refrigerator stores) for the longest two days.
1.2 laboratory animal
18 ~ 22 grams of SPF level NIH healthy mices, provide (mouse conformity certification number: SCXK(Guangdong) 2008-0002 by Guangdong Medical Lab Animal Center).Mouse feeder environment: room temperature 23 ± 2 DEG C, relative humidity 65 ± 10%, illumination every day 12 hours.The full price mouse granulated feed that mouse feed provides for Guangdong Medical Lab Animal Center.
1.3 agents useful for same
Tween-80, Biosharp biotechnology lot number: Amre5000442.
2 methods and result
2.1 experimental technique
Test front 12 h fast (can't help water).NIH healthy mice, random packet, often organizes 10, male and female half and half.According to pre-test result, tested material establishes 5 dosage groups and solvent (the 0.5% tween-80 aqueous solution) control group, and the situation such as dosage and grouping is in table 7, table 8.Each dosage group spacing ratio is 0.7.The administration of mouse single intraperitoneal injection, administration volume is 10ml/kg.Improvement Kou Shifa (Modified Karber ' s method) is adopted to calculate LD 50.
Table 7 multinuclear molecular compound single intraperitoneal injection administration acute toxicity test dead mouse situation
Table 8 multinuclear molecular compound single intraperitoneal injection administration the acute toxicity tests
logLD50=Xm-i（∑P-0.5）
95% fiducial limit=the log-1 (log LD50 ± 1.96 × Sx50) of LD50
Wherein Sx50=i* ( Σp - Σp 2 ) / ( n - 1 )
I-group distance, the i.e. difference of two adjacent groups log10 dose
Xm-maximal dose logarithm
P-each dosage group mortality ratio (mortality ratio represents all decimally)
Pm-most high mortality
Pn-minimum mortality
Σ p-each dosage group mortality ratio sum
N-each treated animal number
The standard error of Sx50-logLD50
2.2 observational technique
The various response situation of initial 4 hours close observation animals after administration, not timing later every day is observed repeatedly (be generally upper and lower noon respectively observe 2 times), Continuous Observation 14 days.The toxic reaction situation of record animal (initial time of toxicity symptom and toxic reaction, severity, time length, whether reversible etc.) and death condition, to to be at death's door and dead animal has carried out naked eyes immediately and cuts open inspection, then do corresponding histopathological examination as found that internal organs are abnormal.
2.3 observation index
Mainly macroscopic outward appearance, behavior (spirit), motion, to breathe and under cardiovascular, Digestive tract and urogenital system function and the concrete observation index such as secretory product, movement be shown in.Weigh appetite and body weight every day, draw body weight gain curve.
(1) hair: whether fluffy agglomerating, matt, contaminated, fall hair, alarm hair;
(2) skin: whether to perspire, subcutaneous whether hemorrhage, fash;
(3) external eyes: secretory product is many, stream is shed tears, blepharoptosis, ophthalmoptosis;
(4) eyeball: whether platycoria, conjunctival congestion, eyelid be with or without sagging, ophthalmoptosis and head tremor;
(5) central nervous system:
1. excitation: activity increases, irritation, dysphoria, come and go run, irritability, bellicose, scream, tremble, jump, faint from fear, fecal incontinence, tetanic;
2. restraining effect: movable to reduce, slow in reacting, One's spirits are drooping, instability of gait, ataxia, disappearance, Curled is sleeping in motionless, the reflection of reposing (pupil, right, stretching, to startle), lethargic sleep, stupor;
(6) somatic movement: whether muscle rigidity, tetraplegia, forced movement, jerk;
(7) to breathe and the recycle system: whether be short of breath, have difficulty in breathing, pant, mouth and nose are hemorrhage, cough, nasal discharge, auricular concha are ice-cold, mucosal pallor, mucous membrane cyanosis, touch pareordia to be to judge heart rate speed;
(8) Digestive tract: whether salivate, abdominal distension, diarrhoea, appetite, stool color (melena, ashen are just) and proterties (rare just, constipation, to have blood in stool);
(9) urogenital system: whether oliguria color depth, blood urine, rhacoma;
(10) other: whether twist one's ears and rub one's face as a sign of anxiety, become thin, fervescence or body temperature reduces.
2.4. experimental result
Vehicle controls group was at the viewing duration of 14 days, and above-mentioned observation index is all normal, and animal, without death, does not occur toxic reaction.
1 hours after tested material multinuclear molecular compound intraperitoneal injection, animal subject starts to occur toxic reaction: walking is waved, and rolls up, burnout, belly indent, expiratory dyspnea.188.7mg/kg dosage group mouse starts to occur death on the 2nd day, and each administration group mouse is most dead in 3 days.The not dead mouse mental status is poor, movable minimizing, and slow in reacting, dorsal body setae is fluffy, matt, recovers gradually after the 6th day.Perform an autopsy on sb. to dead mouse, naked eyes have no the obviously change such as hemorrhage, downright bad of the heart, liver, spleen, lung, kidney, brain.
The observation period of 14 days terminates all mouse of rear execution and performs an autopsy on sb., and each dosage group mice organs has no obvious pathological change.
From experimental result, the LD of multinuclear molecular compound chamber drug administration by injection 50for 279.3mg/kg, multinuclear molecular compound drug administration by injection LD 5095% be crediblely limited to 234.6 ~ 332.5mg.
Experimental result shows that multinuclear molecular compound of the present invention is not obvious for the damage of animal viscera, and toxicity is lower.
Finally to should be noted that; above embodiment is only in order to illustrate technical scheme of the present invention but not limiting the scope of the invention; although be explained in detail the present invention with reference to preferred embodiment; those of ordinary skill in the art is to be understood that; can modify to technical scheme of the present invention or equivalent replacement, and not depart from essence and the scope of technical solution of the present invention.

Claims (16)

1. a multinuclear molecular compound, is characterized in that, described multinuclear molecular compound is synthesized by berberrubine and magnolol, and described multinuclear molecular compound structural formula is:
Described n is the integer of 1≤n≤30.
2. multinuclear molecular compound as claimed in claim 1, it is characterized in that, in the structural formula of described multinuclear molecular compound, n is 2.
3. multinuclear molecular compound as claimed in claim 1, it is characterized in that, described multinuclear molecular compound is prepared from by following methods:
(1a) synthesis of berberrubine derivative: take berberrubine, add organic solvent, adds X (CH after being heated to boiling 2) ny, back flow reaction, concentrates reaction solution, crystallisation by cooling, filters, obtains berberrubine derivative;
(2a) get the berberrubine derivative that step (1a) obtains to mix with magnolol, add anhydrous sodium carbonate and organic solvent, stir, heating reflux reaction, filters reacting liquor while hot, filtrate recycling design, dissolve with DMSO, post is separated, and uses the methanol solution wash-out of 30%, 40%, 50%, 60% successively, HPLC detects, and collects 60% methanol-eluted fractions, concentrated, sample mixed by concentrated solution recycle silicon glue, silicagel column is separated, and wash-out, obtains multinuclear molecular compound;
X (CH described in described step (1a) 2) nin Y, X=Y, X, Y are O, S, F, Cl, Br or I, and n is the integer of 1≤n≤30.
4. multinuclear molecular compound as claimed in claim 3, it is characterized in that, the building-up process of the red alkali derivant of described step (1a) Berberine is: take berberrubine 1 weight part, add organic solvent 150 ~ 200 weight part, 85 DEG C are heated to boiling, add 1, 2-ethylene dibromide 20 ~ 40 weight part, back flow reaction 3h, reaction solution is concentrated into 50 ~ 100 parts, crystallisation by cooling, filter, with appropriate organic solvent washing crystallization, washings and filtrate merge, reclaim organic solvent, the residue dissolve with methanol of 30 weight parts, crystallisation by cooling, filter, use methanol wash crystallization, merge with above-mentioned crystallization, obtain berberrubine derivative.
5. multinuclear molecular compound as claimed in claim 3, it is characterized in that, described step (2a) is: get berberrubine derivative 1.2 weight part that step (1a) obtains and mix with magnolol 1 weight part, add anhydrous sodium carbonate 2 weight part, organic solvent 150 weight part, stir, be heated to 85 DEG C, backflow 8h, reacting liquor while hot is filtered, filtrate recycling design, with the DMSO solvent of 5 weight parts, post is separated, use 30% successively, 40%, 50%, the methanol solution wash-out of 60%, HPLC detects, collect 60% methanol-eluted fractions, concentrated, the silica gel mixed sample of 3 weight parts used again by concentrated solution, silicagel column is separated, with sherwood oil and the ethyl acetate mixtures wash-out of 20 times of column volumes, use methanol-eluted fractions again, reclaim methyl alcohol, obtain multinuclear molecular compound.
6. multinuclear molecular compound as claimed in claim 1, it is characterized in that, described multinuclear molecular compound is prepared from by following methods:
(1b) synthesis of magnolia bark phenol derivative: get magnolol, mix with anhydrous sodium carbonate, add organic solvent, then add X (CH 2) ny, reacting by heating, obtains magnolia bark phenol derivative;
(2b) get magnolia bark phenol derivative that step (1b) obtains, anhydrous sodium carbonate, berberrubine mixing, add organic solvent, stir, heating reflux reaction, filters reacting liquor while hot, filtrate recycling design, dissolve with DMSO, post is separated, and uses the methanol solution wash-out of 30%, 40%, 50%, 60% successively, HPLC detects, and collects 60% methanol-eluted fractions, concentrated, sample mixed by concentrated solution recycle silicon glue, silicagel column is separated, and wash-out, obtains multinuclear molecular compound;
X (CH described in described step (1b) 2) nin Y, X=Y, X, Y are O, S, F, Cl, Br or I, and n is the integer of 1≤n≤30.
7. multinuclear molecular compound as claimed in claim 6, is characterized in that, magnolol, anhydrous sodium carbonate, X (CH in described step (1b) 2) nthe mol ratio of Y is 1:2:16, and described magnolol is 1/150mol/L with the molecular volume ratio of organic solvent, and temperature of reaction is 85 DEG C, and the reaction times is 5h; Described X (CH 2) ny is glycol dibromide.
8. multinuclear molecular compound as claimed in claim 6, it is characterized in that, described step (2b) is: get magnolia bark phenol derivative 1 weight part that step (1b) obtains, anhydrous sodium carbonate 2 weight part, berberrubine 1 weight part adds in reactor, add organic solvent 150 parts, stir, be heated to 85 DEG C, backflow 8h, reacting liquor while hot is filtered, filtrate recycling design, with the DMSO solvent of 5 weight parts, post is separated, use 30% successively, 40%, 50%, the methanol solution wash-out of 60%, HPLC detects, collect 60% methanol-eluted fractions, concentrated, the silica gel mixed sample of 3 weight parts used again by concentrated solution, silicagel column is separated, with sherwood oil and the ethyl acetate mixtures wash-out of 20 times of column volumes, use methanol-eluted fractions again, reclaim methyl alcohol, obtain multinuclear molecular compound.
9. multinuclear molecular compound as claimed in claim 1, is characterized in that, described berberrubine adopts following methods to be prepared from: add Berberine and DMF that solid-liquid ratio is 1:15 ~ 30g/L in the reactor, add zeolite, reflux condensation mode, under 400W ~ 800W microwave radiation, reaction 10 ~ 20min, take out reactor, add water dilution cooling while hot, refrigerated overnight, make crystallization complete, suction filtration, dry, obtain crystallization a; Filtrate is separately separated by macroporous resin column, uses the methanol-eluted fractions of 40%, 45%, 50%, 55%, 60%, 65% and 70% successively, collects 70% methanol-eluted fractions position, concentrated, obtains crystallization b, is merged by crystallization a and crystallization b, obtain berberrubine.
10. the preparation method of the multinuclear molecular compound as described in claim arbitrary in claim 1 ~ 9, is characterized in that, comprise the following steps:
(1a) synthesis of berberrubine derivative: take berberrubine, add organic solvent, adds X (CH after being heated to boiling 2) ny, back flow reaction, concentrates reaction solution, crystallisation by cooling, filters, obtains berberrubine derivative;
(2a) get the berberrubine derivative that step (1a) obtains to mix with magnolol, add anhydrous sodium carbonate and organic solvent, stir, heating reflux reaction, filters reacting liquor while hot, filtrate recycling design, dissolve with DMSO, post is separated, and uses the methanol solution wash-out of 30%, 40%, 50%, 60% successively, HPLC detects, and collects 60% methanol-eluted fractions, concentrated, sample mixed by concentrated solution recycle silicon glue, silicagel column is separated, and wash-out, obtains multinuclear molecular compound;
X (CH described in described step (1a) 2) nin Y, X=Y, X, Y are O, S, F, Cl, Br or I, and n is the integer of 1≤n≤30.
The preparation method of 11. multinuclear molecular compounds as claimed in claim 10, it is characterized in that, the building-up process of the red alkali derivant of described step (1a) Berberine is: take berberrubine 1 weight part, add organic solvent 150 ~ 200 weight part, 85 DEG C are heated to boiling, add 1, 2-ethylene dibromide 20 ~ 40 weight part, back flow reaction 3h, reaction solution is concentrated into 50 ~ 100 parts, crystallisation by cooling, filter, with appropriate organic solvent washing crystallization, washings and filtrate merge, reclaim organic solvent, the residue dissolve with methanol of 30 weight parts, crystallisation by cooling, filter, use methanol wash crystallization, merge with above-mentioned crystallization, obtain berberrubine derivative.
The preparation method of 12. multinuclear molecular compounds as claimed in claim 10, it is characterized in that, described step (2a) is: get berberrubine derivative 1.2 weight part that step (1a) obtains and mix with magnolol 1 weight part, add anhydrous sodium carbonate 2 weight part, organic solvent 150 weight part, stir, be heated to 85 DEG C, backflow 8h, reacting liquor while hot is filtered, filtrate recycling design, with the DMSO solvent of 5 weight parts, post is separated, use 30% successively, 40%, 50%, the methanol solution wash-out of 60%, HPLC detects, collect 60% methanol-eluted fractions, concentrated, the silica gel mixed sample of 3 weight parts used again by concentrated solution, silicagel column is separated, with sherwood oil and the ethyl acetate mixtures wash-out of 20 times of column volumes, use methanol-eluted fractions again, reclaim methyl alcohol, obtain multinuclear molecular compound.
The preparation method of 13. multinuclear molecular compounds as described in claim arbitrary in claim 1 ~ 9, is characterized in that, comprise the following steps:
(1b) synthesis of magnolia bark phenol derivative: get magnolol, mix with anhydrous sodium carbonate, add organic solvent, then add X (CH 2) ny, reacting by heating, obtains magnolia bark phenol derivative;
(2b) get magnolia bark phenol derivative that step (1b) obtains, anhydrous sodium carbonate, berberrubine mixing, add organic solvent, stir, heating reflux reaction, filters reacting liquor while hot, filtrate recycling design, dissolve with DMSO, post is separated, and uses the methanol solution wash-out of 30%, 40%, 50%, 60% successively, HPLC detects, and collects 60% methanol-eluted fractions, concentrated, sample mixed by concentrated solution recycle silicon glue, silicagel column is separated, and wash-out, obtains multinuclear molecular compound;
X (CH described in described step (1b) 2) nin Y, X=Y, X, Y are O, S, F, Cl, Br or I, and n is the integer of 1≤n≤30.
The preparation method of 14. multinuclear molecular compounds as claimed in claim 13, is characterized in that, magnolol, anhydrous sodium carbonate, X (CH in described step (1b) 2) nthe mol ratio of Y is 1:2:8, and described magnolol is 1/150mol/L with the molecular volume ratio of organic solvent, and temperature of reaction is 85 DEG C, and the reaction times is 5h; Described X (CH 2) ny is glycol dibromide.
The preparation method of 15. multinuclear molecular compounds as claimed in claim 13, it is characterized in that, described step (2b) is: get magnolia bark phenol derivative 1 weight part that step (1b) obtains, anhydrous sodium carbonate 2 weight part, berberrubine 1 weight part adds in reactor, add organic solvent 150 parts, stir, be heated to 85 DEG C, backflow 8h, reacting liquor while hot is filtered, filtrate recycling design, with the DMSO solvent of 5 weight parts, post is separated, use 30% successively, 40%, 50%, the methanol solution wash-out of 60%, HPLC detects, collect 60% methanol-eluted fractions, concentrated, the silica gel mixed sample of 3 weight parts used again by concentrated solution, silicagel column is separated, with sherwood oil and the ethyl acetate mixtures wash-out of 20 times of column volumes, use methanol-eluted fractions again, reclaim methyl alcohol, obtain multinuclear molecular compound.
16. 1 kinds as arbitrary in claim 1 ~ 9 as described in multinuclear molecular compound preparation treatment diabetes medicine in purposes.
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