CN104523703A - Application of free fatty acid translocator small-molecule inhibitor - Google Patents
Application of free fatty acid translocator small-molecule inhibitor Download PDFInfo
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Abstract
The invention discloses an application of a free fatty acid translocator small-molecule inhibitor. The free fatty acid translocator small-molecule inhibitor is 6-beta-ethyl-chenodeoxycholic acid or a derivative of 6-beta-ethyl-chenodeoxycholic acid or pharmaceutically acceptable nontoxic salt formed from the derivative; the free fatty acid translocator small-molecule inhibitor has a specific inhibiting effect on free fatty acid translocator 5 (FATP5), and the small-molecule inhibitor can be used for relieving the absorption of free fatty acid in liver and reducing the fatty content of the liver; the small-molecule inhibitor can be used for preparing a medicine for inhibiting the free fatty acid translocator 5 (FATP5) or a medicine for treating nonalcoholic fatty liver disease, and compared to an existing FATP5-inhibiting small-molecule inhibitor, the small-molecule inhibitor disclosed by the invention is better in clinical application effect.
Description
Technical field
The present invention relates to a kind of application of free fatty transport protein micromolecule mortifier, belong to biotechnology and field of medicaments.
Background technology
Non-alcoholic fatty liver disease (NAFLD) has become one of the highest hepatopathy of China's sickness rate.2005, the investigation display NAFLD sickness rate in Shanghai reached 15%.Within 2012, have report display, at the sickness rate of Hong Kong NAFLD up to 27.3%, wherein 3.7% show progressive stage hepatic fibrosis, its degree of cirrhosis and Body Mass Index and transaminase have nothing to do.Along with high fat diet, crowd increases, and pathogenesis of fatty liver rate is also raising year by year.
NAFLD is a kind of multistage chronic disease, with multiple complications:
(1) normally a kind of reversible metabolism syndrome of simple fatty liver is " first strike " that liver is subject to, be after this comparatively vulnerable to medicine, poisonous substance, ethanol, ischemia " second strike ", cause other types hepatopathy; Simple fatty liver occurs because of obesity usually, but its molecular mechanism is failed to understand, so the relation of fatty liver and fat metabolic disturbance still needs further research;
(2) research display, having the simple fatty liver of more than 20% to develop into non-ethanol fat hepatitis, is clearly the important cercinoma prophase pathologic change of cryptogenic cirrhosis and hepatocarcinoma, and molecular mechanism of its development may comprise mitochondrial function exception and inflammatory reaction etc.;
(3) Patients with Fatty Liver hyperlipemia, diabetes and mortality of hypertension increase, and coronary heart disease finally occurs, the probability of apoplexy also can significantly increase, its healthy life span and total life span are subject to the impact of fatty liver.
The treatment of current fatty liver is mainly made the life better mode, perform difficulty, and Europe and U.S. FDA do not ratify the medicine for the treatment of simple fatty liver, so the target spot of the simple fatty liver finding treatment reversible, prevent the generation of follow-up serious disease from having become the vital task of current medical science secondary prevention.In clinical position, partial fat hepatopathy people is due to fat-reducing of going on a diet, and heat Deficiency of Intake in a period of time, also causes fatty liver to increase the weight of.This just points out us, and the major reason of pathogenesis of fatty liver is: liver takes in long-chain fatty acid (Long Chain Fatty Acid, the LCFA) synthctic fat mainly from fatty tissue in blood, and body can not be used.Research shows, serum LCFA raise and fat, insulin resistant is relevant, increase hepatic gluconeogenic and triglyceride generation.LCFA is also the part of nuclear receptor PPAR alfa, beta and delta, affects inflammatory process.Clinically, in Patients with Fatty Liver liver, 60% triglyceride derives from LCFA in blood, and 26% is liver fabricated in situ, and 15% directly from food.Fat content in fat constituent energy rapid contribution liver in food, high fat diet also can cause the fatty liver of people and Mus.As can be seen here, in blood, LCFA is transported into liver is in a large number the important pathological process that fatty liver occurs.
Intestinal, heart, fat, kidney regulating liver-QI etc. have organized long-chain fatty acid transport protein.The more fatty acid transport protein of current research comprises: long-chain fat acid binding protein (FABP), FAT/CD36, ACSL and free fatty transport protein (Free Fatty Acid Transporter, FATP).People or mice FATP family are all made up of 6 albumen, and each albumen of FATP family comprises AMP binding domain, and between albumen, 35% to 50% aminoacid is homologous sequence.That express in liver is FATP2 and FATP5, and its amino acid homology sequence is more than 70%.High expressed liver FATPs enhances the absorption of LCFA.FATP2 has expression liver and kidney, there is long-chain ester acyl CoA synthesizing activity, FATP2 knock-out mice decreases liver and kidney is oxidized the absorption of LCFA and beta, and a kind of neurodegenerative endocrinopathy (ALD) occurs FATP2 knock-out mice, but to knock out be not the immediate cause of this disease to FATP2.FATP5 with FATP2 is the same is also a membrane-spanning protein, and FATP5 only expresses liver, and all show the transport function of LCFA with experiment in vitro in vivo, so FATP5 is a suitable gene knockout target spot.FATP5 knock-out mice decreases liver and takes in LCFA, spontaneously decrease feed food rich in fat, improve the fatty liver of food rich in fat induction, enhance the anabolism of sugar, confirm that FATP5 is one of key factor of the fatty liver formation of Mouse Liver insulin resistant and food rich in fat induction.FATP5 knocks out the composition changing cholic acid in bile, inhibits secondary bile acid and amino acid whose combination, significantly reduces the formation of cholelithiasis.Further experiment builds and purification carries the adeno-associated virus (AAV8) of FATP5 shRNA, finds that striking FATP5 in low liver with this adeno-associated virus expresses, can the prevention and therapy mice food rich in fat fatty liver of inducing.Meanwhile, FATP5 knocks out Old Male mouse adipose liver to be improved, and in blood, testosterone increases, and the conventional FATP5 raised knocks out the male mice life-span and do not have marked difference with the wild-type mice of condition, and FATP5 knocks out and has no chronic side effect.So FATP5 may be the promising target for the treatment of non-ethanol fatty liver.
The micromolecular compound of nearest reported suppression FATP5 comprises chenodeoxycholic acid (CDCA), ursodesoxycholic acid (UDCA), deoxycholic acid (DCA), rarely has the nontoxic salt about 6-beta-ethyl-chenodeoxycholic acid and derivant or derivatives thereof thereof suppressing the relevant report in free fatty transport protein FATP5.
Summary of the invention
The object of this invention is to provide the application of a kind of free fatty transport protein micromolecule mortifier in the medicine of preparation suppression free fatty transport protein FATP5 or the medicine for the treatment of non-alcoholic fatty liver disease, free fatty transport protein micromolecule mortifier of the present invention is the pharmaceutically acceptable nontoxic salt that the derivant or derivatives thereof of 6-beta-ethyl-chenodeoxycholic acid or 6-beta-ethyl-chenodeoxycholic acid is formed, described 6-beta-ethyl-chenodeoxycholic acid structural formula is such as formula shown in IA, and its derivant has such as formula chemical constitution shown in IB:
（IA） （IB）
Wherein, R
1the C1-C5 alkyl of straight or branched, R
2for H, F, Cl, Br, I, CH
3,-CH
2-CH
3one of or derivant shown in formula IB can be made to form other chemical groups of stable meltable salt.
Described nontoxic salts can be: acetas, adipate ester, aspartate, benzoate, benzene sulfonate, bicarbonate/carbonate, disulfate/sulfate, borate, citrate, cyclohexyl-n-sulfonate, esilate, formates, fumarate, gluceptate, gluconate, glucuronate, hexafluorophosphate, hydrochlorate/chloride, hydrobromate, hydrophosphate, bromide, hydriodate/iodide, isethionate, D-and Pfansteihl salt, malate, maleate, malonate, mesylate, 2-naphthalene sulfonate, nicotinate, nitrate, Orotate, oxalates, palmitate, embonate, phosphate, dihydric phosphate, pyroglutamate, sucrose, stearate, succinate, tannin, D-and L-TARTARIC ACID salt, one in 1-hydroxyl-2-naphthalene tosylate or xinafoate.
Described nontoxic salts can also be react by the derivant and one of following ion or material with described formula IB structure the pharmaceutically acceptable salt formed: (1) and Al
3+, Ca
2+, Mg
2+, Zn
2+, Na
+, K
+the basic salt prepared; (2) salt formed with the amine such as arginine, glycine, lysine; (3) and N, N '-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine, procaine etc. formed organic salt.
Described pharmaceutical dosage form can be tablet, injection, oral liquid, capsule, patch, spray or the other drug dosage form that medically accepts.
Described suppression free fatty transport protein micromolecule mortifier can also be applied to preparation obesity, metabolism syndrome, insulin resistant, diabetes, cardiovascular diseases, the medicine for treatment of apoplexy or adjuvant drug.
The present invention also provides with following three kinds be main active with described free fatty transport protein micromolecule mortifier 6-beta-ethyl-chenodeoxycholic acid the medicine of suppression free fatty transport protein FATP5 or medicines for the treatment of non-alcoholic fatty liver disease.
Treat a medicine for non-alcoholic fatty liver disease, be made up of free fatty transport protein micromolecule mortifier 6-beta-ethyl-chenodeoxycholic acid and the adjuvant that pharmaceutically accepts, adopt following concrete steps to prepare:
(1) preparation prescription: 6-beta-ethyl-chenodeoxycholic acid 100g, microcrystalline Cellulose 240g, soluble starch 150g, Pulvis Talci 10g;
(2) take each raw material by preparation prescription, 6-beta-ethyl-chenodeoxycholic acid and soluble starch are mixed, then adds microcrystalline Cellulose, with 95% ethanol soft material, to sieve granulation, at low temperature 50 DEG C of dryings, then granulate, add Pulvis Talci, mixing, tabletting, every sheet 0.5g, quality inspection, packaging, obtain the tablet of every sheet containing 6-beta-ethyl-chenodeoxycholic acid 100mg.
Suppress a medicine of free fatty transport protein FATP5, be made up of free fatty transport protein micromolecule mortifier 6-beta-ethyl-chenodeoxycholic acid and the adjuvant that pharmaceutically accepts, adopt following concrete steps to prepare:
(1) preparation prescription: core: 6-beta-ethyl-chenodeoxycholic acid 100g, soybean oil 300g, capsule material: gelatin-glycerol-water 100g, the weight ratio of gelatin, glycerol, water is 1: 0.4: 1, makes 1000 soft capsules;
(2) getting 6-beta-ethyl-chenodeoxycholic acid adds in soybean oil, stirs evenly and makes dissolving, on encapsulating machine, press capsule, dry, packaging, obtains the soft capsule of every ball containing the 6-beta-ethyl-chenodeoxycholic acid of 100mg.
Suppress a medicine of free fatty transport protein FATP5, be made up of free fatty transport protein micromolecule mortifier 6-beta-ethyl-chenodeoxycholic acid and the adjuvant that pharmaceutically accepts, adopt following concrete steps to prepare:
(1) preparation prescription: 6-beta-ethyl-chenodeoxycholic acid 10g, soybean oil 250g, soybean lecithin 10g, poloxamer 60g, sodium benzoate 4g, aspartame 3g, distilled water surplus is to 1000ml;
(2) preparation method: 6-beta-ethyl-chenodeoxycholic acid and soybean lecithin are added in soybean oil, oil phase is obtained after abundant dissolving, poloxamer, sodium benzoate and aspartame are dissolved in 500ml water and obtain aqueous phase, under high shear dispersion dispersing emulsification machine stirs, aqueous phase is slowly added in oil phase, 6000rpm stirs 15min and obtains colostrum, add water to 1000ml, circulate 5 times through high pressure homogenizer, obtain milky breast eventually, fill nitrogen fill, obtain the Orally taken emulsion that every mL contains 10mg 6-beta-ethyl-chenodeoxycholic acid.
The preparation method of micromolecule mortifier of the present invention, synthesizes example with 6-beta-ethyl-chenodeoxycholic acid (formula IA), adopts following steps:
(1) at 30 ~ 60 DEG C, in methanol solution, methanesulfonic acid esterification 3 Alpha-hydroxy-7-ketone group-5 β-cholanic acid (II) is used, to obtain 3 Alpha-hydroxy-7-ketone group-5 β-cholanic acid methyl ester (III);
(2) under tertiary amine that is aliphatic, alicyclic or heteroaromatic type exists, in aromatic solvent, use trim,ethylchlorosilane by-5 β-cholanic acid methyl ester (III) silylanizing of 3-Alpha-hydroxy-7-ketone group, to obtain corresponding 3-α-trimethylsiloxy-7-ketone group-5 β-cholanic acid methyl ester (IV);
(3) under the alkaline amide of the alkaline amide obtained by ammonia or aliphatic secondary amine and trim,ethylchlorosilane exist, by 3 α-trimethylsiloxy-7-ketone group-5 β-cholanic acid methyl ester (IV) silylanizing in the aprotic solvent of polarity, with obtain 3 α-, 7-bis--trimethylsiloxy-6-alkene-5 β-cholanic acid methyl ester (V);
(4) at-90 ~-60 DEG C, make 3-α-, 7-bis--trimethylsiloxy-6-alkene-5 β-cholanic acid methyl ester (V) and wherein R
1the aldehyde R of the C1-C5 alkyl of straight or branched
1-CHO reacts 2 to 4 hours in alkyl halide solvents and etherate of trifluoroboron, subsequently, reactant mixture is incubated 1 to 6 hour at 0 ~ 35 DEG C, to obtain 3-Alpha-hydroxy-6-alkylidene-7-ketone group-5 β-cholanic acid methyl ester (VI);
(5) at 20 ~ 60 DEG C, under alkaline hydrated oxide exist, in alcoholic solvent, be hydrolyzed 3-Alpha-hydroxy-6-ethylidene-7-ketone group-5 β-cholanic acid methyl ester (VI), obtain 3-Alpha-hydroxy-6-alkylidene-7-ketone group-5 β-cholanic acid (VII),
(6) under 1 ~ 3 atmospheric pressure, in sodium hydrate aqueous solution, use Pd/C hydrogenation 3-Alpha-hydroxy-6-alkylidene-7-ketone group-5 β-cholanic acid, obtain 3-Alpha-hydroxy-6 β-alkyl-7-ketone group-5 β-cholanic acid (VIII);
(7) at 70 ~ 105 DEG C, in the aqueous solution of alkalescence, use metal hydride reduction 3 Alpha-hydroxy-6 β-alkyl-7-ketone group-5 β-cholanic acid (VIII) 1 hour, with obtain 3 α-, 7 alpha-dihydroxy--6 β-alkyl-5 β-cholanic acid (IA),
Concrete synthetic route is as follows:
。
The present invention is as follows relative to the beneficial effect of prior art: the present inventor finds in research process, bile acid activates FXR(method Buddhist nun ester derivant X receptor) regulate and control the absorption process of liver long-chain fatty acid LCFA afterwards, the obesity suppressing intestinal FXR can treat high lipid food to cause.CDCA(chenodeoxycholic acid) be the activator of FXR, and also the derivant that the hydrophobic fat chain group of the 6 α positions of CDCA substitutes generation can strengthen the exciting function of FXR; DCA(deoxycholic acid) the same with LCA, also have activated FXR, but LCA fails to improve fatty liver.In addition, 6-alpha-ethyl-chenodeoxycholic acid is the potent activator of FXR, it is 99nM that the half of 6-alpha-ethyl-CDCA acid active FXR outside born of the same parents activates concentration (EC50), and in cell, EC50 is 85nM, but finds unsatisfactory curative effect in II clinical trial phase.The hepatoma H22 cells of the FXR positive or primary hepatocyte is utilized to measure, find that FXR agonist 6-alpha-ethyl-chenodeoxycholic acid can not suppress the transport function of FATP5 when low concentration 100nM, but have activated FXR, the absorption significantly increasing the primary hepatocyte LCFA of the FATP5 positive in 30 minutes reaches 52%, about 5% is strengthened to the absorption of the HepG2 cell LCFA not expressing FATP5, but 52% is reached to the primary hepatocyte absorption LCFA increase expressing FATP5.Can think that the major reason that 6-alpha-ethyl-chenodeoxycholic acid fails to succeed in clinical studies adds liver after it activates FXR to take in LCFA.
Based on above-mentioned cognition, the present invention is through high flux screening, cytologic experiment, Experiment of Zoology and pharmacodynamics test find, relative to existing FATP5 inhibitor, 6-beta-ethyl-chenodeoxycholic acid is the potent inhibitor of FATP5, the IC50 of primary hepatocyte is about 10nM, its EC50 activating FXR is 16 μMs, because in cell, useful effect concentration differs 1600 times, in its effective suppression body during FATP5 effect, the expression of FXR can not be activated, improve fatty liver, thus reduce serum high-density LP (HDL) after avoiding 6-alpha-ethyl-CDCA acid active liver F XR, increase the side effect of serum low-density LP (LDL), the obesity that may cause after also avoiding sustained activation small intestinal FXR.The nontoxic salts that such free fatty transport protein micromolecule mortifier 6-beta-ethyl-chenodeoxycholic acid, its derivant and derivant thereof are formed is to free fatty transport protein 5(FATP5) suppress there is specificity, liver free fatty can be reduced absorb, reduce hepatic fat content, can be used for primary cell, cell strain, stable cell line and various tissue, if liver, kidney etc. are to the suppression of fatty acid absorption, also comprise the absorption of the fatty acid in food.The nontoxic salts such free fatty transport protein micromolecule mortifier 6-beta-ethyl-chenodeoxycholic acid, its derivant and derivant thereof formed is for the preparation of the medicine of the medicine or treatment non-alcoholic fatty liver disease that suppress free fatty transport protein FATP5, relative to the micromolecule mortifier of existing FATP5, there is larger clinical value.
Accompanying drawing explanation
Fig. 1 is the half-inhibition concentration (IC50) that experiment one measures that in 293-FATP5 cell 6-beta-ethyl-chenodeoxycholic acid suppression FATP5 absorbs long-chain fatty acid.
Fig. 2 is that after experiment two lumbar injection 6-beta-ethyl-chenodeoxycholic acid, fluorescently-labeled fatty acid absorption enters blood and has no appreciable impact, but significantly suppress it and be absorbed into liver.
Fig. 3 is after experiment three human primary hepatocyte and 6-beta-ethyl-chenodeoxycholic acid, fluorescent labeling fatty acid hatch 30 minutes altogether, and 6-beta-ethyl-chenodeoxycholic acid significantly suppresses human primary hepatocyte absorption fatty acid to enter in cell.
Detailed description of the invention
Be described in further details the present invention below by embodiment, these embodiments are only used for the present invention is described, do not limit the scope of the invention.
Embodiment 1 free fatty transport protein micromolecule mortifier is to the suppression of free fatty transport protein
Test the high flux screening of a free fatty transport protein micromolecular inhibitor and the half-inhibition concentration to FATP5 thereof.
1, the stable cell line of expressing different FATP is respectively built: by 1 × 10
4individual human embryo kidney 293 cells is inoculated in 96 orifice plates, when its fusion reaches 30% to 50%, MOI is 10, respectively with the recombinant slow virus and the contrast virus infected cell that carry the FATP such as hsFATP5 or hsFATP2, add virus transfection reinforcing agent polybrene(polybrene), final concentration is 6ug/ml, removes virus liquid after 8 hours, renew fresh culture medium, amplification cultivation also goes down to posterity to build and is.
2, the different FATP of Real_time quantitative detection high expressed absorbs the speed of fatty acid: by 30,000 – 50,293 stabilized cells or the primary hepatocyte of the different FATP of 000 cells/well high expressed are inoculated in 96 orifice plates, every hole 100ul DMEM/FBS, overnight incubation, grow to 50%-70%, then with the 1 × HBSS dilution containing 20mM HEPES, 4.5g/L glucose, 0.1%BSA.Again with containing or not adding 96 orifice plates and 24 orifice plates respectively containing the HBSS BSA buffer of 100 μ l or 1ml of the micromolecule mortifier 6-beta-ethyl-chenodeoxycholic acid of free fatty transport protein, hatch 30min, then remove buffer.
3, inhibitor screening experiment: add the BODIPY-FA mixture (containing the HBSS of 4.5g/L glucose, 0.1% BSA, 2 μMs of fluorescently-labeled LCFA C1-BODIPY-C12,2mM quenchers) containing 6-beta-ethyl-chenodeoxycholic acid.Put on real-time continuous fluorescence plate reading machine (Gemini plate reader) after adding these mixture at once, plate reading machine is set to bottom and reads, high responsive, excitation wavelength 488nm, emission wavelength 515nm, can read once every 2min, altogether 2h, so only detect the fluorescence of adherent cell, because block the fluorescence in mixture with quencher.Absorption speed carries out linear regression calculating by the data point detected.
4, the high flux screening of free fatty transport protein micromolecule mortifier: 56 kinds of different bile acid derivatives are dissolved in DMSO respectively as storage liquid, add above BODIPY-FA mixture, dilution bile acid derivative is the final concentration needed, add in order to upper method in the cell of the different FATP of high expressed, put into plate reading machine at once and detect fatty acid absorption speed.The reversibility of simultaneously carrying out bile acid derivative suppression detects: wash twice cell with PBS, then adds not containing the BODIPY-FA quencher mixture of bile acid derivative, and the IC50 of compound, by statistics, calculates by nonlinear regression.
The half-inhibition concentration (IC50) of result display section bile acid derivative to FATP5 is as shown in table 1: 6-beta-ethyl-chenodeoxycholic acid (Fig. 1) IC50 is less than 1nM.
The FATP5 mortifier that table 1 screens and its IC50
。
Test the suppression that in two: hours, 6-beta-ethyl-chenodeoxycholic acid absorbs mouse liver long-chain fatty acid (LCFA).
Mice fasting one evening, the 2nd day morning 10 point, anesthetized mice.With the PBS liquid lumbar injection of the fluorescently-labeled fatty acid of the C1-BODIPY-C12 of 20-100 μM in conjunction with 1% BSA.The 6-beta-be dissolved in DMSO ethyl-chenodeoxycholic acid is added in PBS, injection opposite side abdominal cavity.Put to death mice after one hour, get serum and liver, liver makes albumen homogenate after weighing in 2ml RIPA buffer, measures protein concentration, fat-soluble LCFA is extracted with the Dole reagent (petroleum ether: isopropyl alcohol: 2N sulphuric acid=10:40:1, volume ratio) of 3 times of volumes.With 18000g rotating speed centrifugal 10 minutes, collect supernatant.Get 50ul supernatant or serum adds 96 orifice plates, with excitation wavelength 488nm, emission wavelength 515nm measures fluorescence, and flat fluorescent protein concentration or organ weight carry out standardization.
Result as shown in Figure 2,3, result shows: 6-beta-ethyl-chenodeoxycholic acid did not make significant difference (Fig. 2 b) to content of fatty acid in serum in one hour, but (Fig. 2 a), to absorb fatty acid to liver inhibited for prompting 6-beta-ethyl-chenodeoxycholic acid to significantly suppress the content of fluorescent labeling fatty acid in liver.
Experiment three: 6-beta-ethyl-chenodeoxycholic acid absorbs the suppression of long-chain fatty acid (LCFA) to human primary hepatocyte
Be separated human primary hepatocyte, be suspended in HBSS, the 6-beta-of the 10-50nM in DMSO ethyl-chenodeoxycholic acid is added the HBSS of the BODIPY-LCFA combined containing 0.1%BSA, then add in cell, at 37 ° of C, 5% CO
2in hatch 30 minutes, be placed on ice, before stream measuring, add PI.Flow cytometer is positive and negative identification living cells and dead cell by PI, detects LCFA take in situation by detecting BODIPY green fluorescence intensity.
Result shows: 6-beta-ethyl-chenodeoxycholic acid inhibits the absorption of LCFA, but the hepatocyte of FATP5 knock-out mice can not be suppressed to absorb LCFA, and primary hepatocyte (PI negative cells) alive also can not be caused to decline.Trypan Blue detects primary hepatocyte, its live cell fraction and do not add 6-beta-ethyl-chenodeoxycholic acid matched group without significant difference (Fig. 3).
Embodiment 2 6-beta-ethyl-chenodeoxycholic acid is as follows for the preparation of the applicating example suppressed in the medicine of the medicine of free fatty transport protein FATP5 or treatment non-alcoholic fatty liver disease.
One, tablet applications.
Preparation prescription: 6-beta-ethyl-chenodeoxycholic acid 100g, microcrystalline Cellulose 240g, soluble starch 150g, Pulvis Talci 10g; Make 1000 altogether, every sheet is containing 6-beta-ethyl-chenodeoxycholic acid 100 mg.
Preparation method: take 6-beta-ethyl-chenodeoxycholic acid 100 g, with soluble starch 150g, mixing, take microcrystalline Cellulose 240g again, with 95% ethanol soft material, granulation of sieving, low temperature 50 DEG C of dryings, granulate, add Pulvis Talci 10g, mixing, tabletting (every 0.5g), quality inspection, packaging, to obtain final product.
Usage and dosage: every day 4-8 sheet, point 2-3 time.
The crowd of being suitable for: be suitable for nonalcoholic fatty liver disease, high-fat animal food of especially taking food is main patient.
Two: the preparation of soft capsule.
Prescription: core: 6-beta-ethyl-chenodeoxycholic acid 100g, soybean oil 300g, capsule material: gelatin-glycerol-water (weight ratio between three is 1: 0.4: 1) 100g, makes 1000 soft capsules.
Preparation method: get 6-beta-ethyl-chenodeoxycholic acid and add in soybean oil, stir evenly and make dissolving, press capsule on encapsulating machine, dry, packaging, obtains the soft capsule of every ball containing the 6-beta-ethyl-chenodeoxycholic acid of 100mg.Oral: 400-800mg/ day, i.e. 4-8 ball/day, divide and take for 2-3 time.
Three: the preparation of Orally taken emulsion.
Prescription: 6-beta-ethyl-chenodeoxycholic acid 10g, soybean oil 250g, soybean lecithin 10g, poloxamer 60g, sodium benzoate 4g, aspartame 3g, distilled water adds to 1000ml.
Preparation method: 6-beta-ethyl-chenodeoxycholic acid and soybean lecithin are added in soybean oil, oil phase is obtained after abundant dissolving, poloxamer, sodium benzoate and aspartame are dissolved in 500ml water and obtain aqueous phase, slowly added in oil phase by aqueous phase under high shear dispersion dispersing emulsification machine stirs, 6000rpm stirs 15min and obtains colostrum, adds water to 1000ml, circulate 5 times through high pressure homogenizer, obtain milky breast eventually, fill nitrogen fill, obtain 6-beta-ethyl-chenodeoxycholic acid that every mL contains 10mg.
Oral: 400-800mg/ day, i.e. 40-80ml/ day, divide and take for 2-3 time, note during use shaking up.
Claims (8)
1. the application of a free fatty transport protein micromolecule mortifier, it is characterized in that: for the preparation of the medicine of the medicine or treatment non-alcoholic fatty liver disease that suppress free fatty transport protein FATP5, the pharmaceutically acceptable nontoxic salts that described micromolecule mortifier is 6-beta-ethyl-chenodeoxycholic acid, its derivant or derivatives thereof is formed, 6-beta-ethyl-chenodeoxycholic acid structural formula is such as formula shown in IA, and its derivant has such as formula chemical constitution shown in IB:
（IA） （IB）
Wherein, R
1for the C1-C5 alkyl of straight or branched, R
2for H, F, Cl, Br, I, CH
3,-CH
2-CH
3one of or derivant shown in formula IB can be made to form other chemical groups of stable meltable salt.
2. application according to claim 1, it is characterized in that: described nontoxic salts comprises: acetas, adipate ester, aspartate, benzoate, benzene sulfonate, bicarbonate/carbonate, disulfate/sulfate, borate, citrate, cyclohexyl-n-sulfonate, esilate, formates, fumarate, gluceptate, gluconate, glucuronate, hexafluorophosphate, hydrochlorate/chloride, hydrobromate, hydrophosphate, bromide, hydriodate/iodide, isethionate, D-and Pfansteihl salt, malate, maleate, malonate, mesylate, 2-naphthalene sulfonate, nicotinate, nitrate, Orotate, oxalates, palmitate, embonate, phosphate, dihydric phosphate, pyroglutamate, sucrose, stearate, succinate, tannin, D-and L-TARTARIC ACID salt, one in 1-hydroxyl-2-naphthalene tosylate or xinafoate.
3. application according to claim 1, is characterized in that: described nontoxic salts is have the derivant of described formula IB structure and one of following ion or material react the pharmaceutically acceptable salt formed: Al
3+, Ca
2+, Mg
2+, Zn
2+, Na
+, K
+, arginine, glycine, lysine, N, N '-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine, procaine.
4. application according to claim 1, is characterized in that: described medicine is tablet, injection, oral liquid, capsule, patch, spray or the other drug dosage form that medically accepts.
5. treat the medicine of non-alcoholic fatty liver disease for one kind, it is characterized in that: be made up of free fatty transport protein micromolecule mortifier 6-beta-ethyl-chenodeoxycholic acid according to claim 1 and the adjuvant that pharmaceutically accepts, its preparation method comprises following concrete steps:
(1) preparation prescription: 6-beta-ethyl-chenodeoxycholic acid 100g, microcrystalline Cellulose 240g, soluble starch 150g, Pulvis Talci 10g;
(2) take each raw material by preparation prescription, 6-beta-ethyl-chenodeoxycholic acid and soluble starch are mixed, then adds microcrystalline Cellulose, with 95% ethanol soft material, to sieve granulation, at low temperature 50 DEG C of dryings, then granulate, add Pulvis Talci, mixing, tabletting, every sheet 0.5g, quality inspection, packaging, obtain the tablet of every sheet containing 6-beta-ethyl-chenodeoxycholic acid 100mg.
6. one kind is suppressed the medicine of free fatty transport protein FATP5, it is characterized in that: be made up of free fatty transport protein micromolecule mortifier 6-beta-ethyl-chenodeoxycholic acid according to claim 1 and the adjuvant that pharmaceutically accepts, its preparation method comprises following concrete steps:
(1) preparation prescription: core: 6-beta-ethyl-chenodeoxycholic acid 100g, soybean oil 300g, capsule material: gelatin-glycerol-water 100g, the weight ratio of gelatin, glycerol, water is 1: 0.4: 1, makes 1000 soft capsules;
(2) getting 6-beta-ethyl-chenodeoxycholic acid adds in soybean oil, stirs evenly and makes dissolving, on encapsulating machine, press capsule, dry, packaging, obtains the soft capsule of every ball containing the 6-beta-ethyl-chenodeoxycholic acid of 100mg.
7. one kind is suppressed the medicine of free fatty transport protein FATP5, it is characterized in that: be made up of free fatty transport protein micromolecule mortifier 6-beta-ethyl-chenodeoxycholic acid according to claim 1 and the adjuvant that pharmaceutically accepts, its preparation method comprises following concrete steps:
(1) preparation prescription: 6-beta-ethyl-chenodeoxycholic acid 10g, soybean oil 250g, soybean lecithin 10g, poloxamer 60g, sodium benzoate 4g, aspartame 3g, distilled water surplus is to 1000ml.
8.(2) preparation method: 6-beta-ethyl-chenodeoxycholic acid and soybean lecithin are added in soybean oil, oil phase is obtained after abundant dissolving, poloxamer, sodium benzoate and aspartame are dissolved in 500ml water and obtain aqueous phase, under high shear dispersion dispersing emulsification machine stirs, aqueous phase is slowly added in oil phase, 6000rpm stirs 15min and obtains colostrum, add water to 1000ml, circulate 5 times through high pressure homogenizer, obtain milky breast eventually, fill nitrogen fill, obtain the Orally taken emulsion that every mL contains 10mg 6-beta-ethyl-chenodeoxycholic acid.
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