CN103172693B - A kind of sterol derivative, Preparation Method And The Use - Google Patents
A kind of sterol derivative, Preparation Method And The Use Download PDFInfo
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- CN103172693B CN103172693B CN201110441993.2A CN201110441993A CN103172693B CN 103172693 B CN103172693 B CN 103172693B CN 201110441993 A CN201110441993 A CN 201110441993A CN 103172693 B CN103172693 B CN 103172693B
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Abstract
The invention belongs to field of medicine and chemical technology, relate to a kind of sterol derivative, Preparation Method And The Use.Particularly, the present invention relates to the compound shown in formula I, or its pharmacy acceptable salt, wherein, R
1be selected from-OH ,=O, H and C
1-C
3alkyl; R
2be selected from-OH, H and C
1-C
3alkyl; R
3be selected from-OH ,=O, H and C
1-C
3alkyl; R
4be selected from-OH, H and C
1-C
3alkyl; And R
1, R
2, R
3, and R
4in any 2,3 or 4 be-OH simultaneously.Compound of the present invention can suppress HMG-CoA reductase effectively, has as reducing or adjusting blood lipid or prevent and/or treat the potentiality of hyperlipemia, hyperlipidemia, hypercholesterolemia or atherosclerotic medicine.
Description
Technical field
The invention belongs to field of medicine and chemical technology, relate to a kind of sterol derivative, Preparation Method And The Use.
Background technology
Red colouring agent for food, also used as a Chinese medicine (Monascus-fermentedrice) take rice as raw material, and a kind of red-purple rice made through monascus (Monascus) fermentation is bent.Red colouring agent for food, also used as a Chinese medicine to claim red bent ancient times, and be that the female or distiller's yeast of main song is inoculated in rice top fermentation and forms by monascus, its look crimson, therefore have another name called red song, red rice, red rice, red wine dregs, again because main product is in Fujian and other places, therefore has another name called good fortune song, good fortune rice etc.
China utilizes the with a long history of Monascus anka Nakazawa et sato, from Han dynasty, just use its koji.Red colouring agent for food, also used as a Chinese medicine is the traditional Chinese medicine that dietotherapy is simply had both.Early it has been widely used in food color, wine brewing, fermentation, traditional Chinese medicine aspect in ancient times.Principle of Correct Diet has red colouring agent for food, also used as a Chinese medicine " taste is sweet, flat, nontoxic ", " in invigorating the spleen, beneficial gas, temperature "; Compendium of Material Medica has " sweet, warm, nontoxic ", " control woman's blood pain and postpartum extravesated blood clean, beat the good of beverage "; Records such as " invigorate blood circulation, help digestion, stomach is warmed up in invigorating the spleen, control red white diarrhea, wound " that " Amplification on Materia Medica addendum " has.
The seventies in last century, since Japan professor Endo isolates physiologically active substance Mo Nakelin K (monacolinK) first from red monascus (Monascusruber), numerous Chinese scholars constantly finds physiologically active substance in monascus meta-bolites, comprise monacolin compounds, monascorubin, some terpenoids of antihypertensive compositions GABA and antioxidant component dimerumicacid and separation recently etc.Along with modern biochemistry and pharmacological development, the reducing blood-fat of red colouring agent for food, also used as a Chinese medicine, step-down, hypoglycemic, anti-obesity, anticancer, effect such as control senile dementia and osteoporosis etc. are constantly excavated.Thus add new intension for traditional red colouring agent for food, also used as a Chinese medicine.
XUEZHIKANG JIAONANG adjusts fat Chinese medicine at efficient, the safe domestic modern times that the mould fermentation of purpose-made monascus that Beijing WBL Peking University Biotech Co., Ltd is independently studied is made.Indication is the treatment of the cardiovascular and cerebrovascular diseases that hyperlipidaemia and atherosclerosis cause.Its main mechanism is the activity by suppressing 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase enzyme, the synthesis of endogenous cholesterol can be suppressed, reduce serum total cholesterol (TC), blood triglyceride (TG), low-density lipoprotein (LDL), high density lipoprotein increasing (HDL) level.XUEZHIKANG JIAONANG is the alcohol extract of red colouring agent for food, also used as a Chinese medicine, and it is rich in a series of natural statin compounds such as lovastatin.Except statin compound, also containing pigment compound, isoflavonoid, phytosterin compound, 20 seed amino acids, unsaturated fatty acids and various trace elements in XUEZHIKANG JIAONANG.Therefore still need and will find the new compound that there is lipopenicillinase or suppress HMG-CoA reductase activity.
Summary of the invention
The present inventor is through deep research and performing creative labour, obtain a kind of sterol derivative, and the present inventor is surprised to find, compound of the present invention can suppress HMG-CoA reductase effectively, thus has as reducing or adjusting blood lipid or prevent and/or treat the potentiality of hyperlipemia, hyperlipidemia, hypercholesterolemia or atherosclerotic medicine.Thus provide following invention:
One aspect of the present invention relates to the compound shown in formula I, or its pharmacy acceptable salt,
Wherein,
R
1be selected from-OH ,=O (carbonyl), H and C
1-C
3alkyl;
R
2be selected from-OH, H and C
1-C
3alkyl;
R
3be selected from-OH ,=O, H and C
1-C
3alkyl;
R
4be selected from-OH, H and C
1-C
3alkyl;
And R
1, R
2, R
3, and R
4in any 2,3 or 4 be-OH simultaneously.
In one embodiment of the invention, R
1for-OH or=O (carbonyl), R
2, R
3with
R
4be H.
Compound according to any one of the present invention or its pharmacy acceptable salt, it is the compound shown in formula II below, or its pharmacy acceptable salt,
The chemistry of formula II compound is called: 16,22-epoxy ergot steroid-5,7-diene-3,20,23,25-tetrol (16,22-epoxy-ergosta-5,7-dien-3,20,23,25-tetraol).
The invention still further relates to hydrate or the solvate of above-mentioned formula I or formula II compound.
Another aspect of the present invention relates to the preparation method of formula I or formula II compound, comprises the steps:
1) alcohol extract (the such as XUEZHIKANG JIAONANG content of red colouring agent for food, also used as a Chinese medicine and/or red colouring agent for food, also used as a Chinese medicine is got, it is dry powder), one or many is extracted as solvent supersonic with one or more organic solvents be selected from methylene dichloride, ethyl acetate, acetone, methyl alcohol, ethanol of 2-6 times of volume, each 20-40 minute, united extraction liquid, except desolventizing, obtain extract;
Alternatively, described alcohol extract can obtain in the following manner: extract one or many as solvent supersonic, each 20-40 minute, united extraction liquid with the 50%-100% ethanol of 2-6 times of volume or 50%-100% methyl alcohol, except desolventizing, obtain alcohol extract.
Be not limited to theoretical restriction, because Xuezhikang itself is the alcohol extract of red colouring agent for food, also used as a Chinese medicine, can be directly therefore raw material with red colouring agent for food, also used as a Chinese medicine, extraction step be substantially identical with XUEZHIKANG JIAONANG content dry powder, but the content of this compound in red colouring agent for food, also used as a Chinese medicine is relatively low.The concrete bacterial strain of described red colouring agent for food, also used as a Chinese medicine is not particularly limited, and comprises the arbitrary bacterial classification or the bacterial strain that belong to red colouring agent for food, also used as a Chinese medicine.XUEZHIKANG JIAONANG (such as Beijing University's dimension letter is produced) can be bought by hospital or pharmacy and obtain.
2) by step 1) extract that obtains carries out silica gel column chromatography separation, carries out gradient elution by sherwood oil and ethyl acetate; The volume ratio of petroleum ether-ethyl acetate is followed successively by 75: 25,50: 50-25: 75,0: 100;
3) get step 2) in petroleum ether-ethyl acetate be the elution fraction of 50: 50-25: 75, be separated through C18 reversed-phase column chromatography, carry out gradient elution with methanol-water, the volume ratio of methanol-water is followed successively by 10: 90,50: 50-75: 25,100: 0;
4) get step 3) in methanol-water be 50: 50-75: 25 elution fraction purify with half preparative high-performance liquid chromatographic, with acetonitrile-0.2% acetic acid aqueous solution (45: 55) for moving phase, C18 semipreparative column is stationary phase, collects the chromatographic peak part of 9.2min; With
5) by step 4) product carry out lyophilize, obtain formula I or formula II compound.
Preparation method according to any one of the present invention, it meets any one in following (1)-(9) or multinomial:
(1) step 1) in, described organic solvent is preferably methylene dichloride;
(2) step 1) in, described supersound extraction carries out 3 times;
(3) step 1) in, by concentrating under reduced pressure except desolventizing;
(4) step 2) in, the volume ratio of petroleum ether-ethyl acetate is followed successively by 75: 25,50: 50,25: 75,0: 100;
(5) step 3) in, get step 2) in petroleum ether-ethyl acetate be the elution fraction of 50: 50 or 25: 75;
The present inventor by test find, petroleum ether-ethyl acetate be 50: 50-25: 75 elution fraction all contain compound of the present invention.
(6) step 3) in, the volume ratio of methanol-water is followed successively by 10: 90,50: 50,75: 25,100: 0;
(7) step 4) in, get step 3) in methanol-water be the elution fraction of 50: 50 or 75: 25;
The present inventor by test find, methanol-water be 50: 50-75: 25 elution fraction all contain compound of the present invention.
(8) step 4) in, the chromatographic peak part of the 9.2min collected is merged; With
(9) step 5) in, cryodesiccated condition is: condenser temperature-40 to-85 DEG C, vacuum tightness 0-100Pa; Be preferably condenser temperature-50 to-82.7 DEG C, vacuum tightness 2-13Pa; Be more preferably condenser temperature-82.7 DEG C, vacuum tightness 2Pa, or condenser temperature-50 DEG C, vacuum tightness 8.5Pa.
Another aspect of the invention relates to a kind of extract, and it contains formula II compound of the present invention.
Extract according to any one of the present invention, it is the extract of XUEZHIKANG JIAONANG content dry powder or the extract of red colouring agent for food, also used as a Chinese medicine.
Extract according to any one of the present invention, its be following (1) to any one in (3):
(1) above-mentioned step 1) to 2) in obtained petroleum ether-ethyl acetate be the elution fraction of 50: 50-25: 75; Preferred petroleum ether-ethyl acetate is the elution fraction of 50: 50 or 25: 75; Preferably petroleum ether-ethyl acetate is the elution fraction of 25: 75 further.
(2) above-mentioned step 1) to 3) in obtained methanol-water be the elution fraction of 50: 50-75: 25; Particular methanol-water is the elution fraction of 50: 50 or 75: 25; Further particular methanol-water is the elution fraction of 75: 25; With
(3) above-mentioned step 1) to 4) in the chromatographic peak part of obtained 9.2min.
Another aspect of the invention relates to a kind of composition, the compound of its contained I or formula II and/or the extract according to any one of the present invention; Alternatively, pharmaceutically acceptable carrier or auxiliary material is also comprised.Particularly, described composition is pharmaceutical composition.
Another aspect of the invention relates to compound of the present invention or the extract described in any one of the present invention or composition of the present invention and reduces or adjusting blood lipid or the purposes that prevents and/or treats in hyperlipemia, hyperlipidemia, hypercholesterolemia or atherosclerotic medicine in preparation.
Usual pharmaceutical composition of the present invention contains the formula I of 0.1-90 % by weight or formula II compound and/or its physiologically acceptable salt.Pharmaceutical composition can be prepared according to methods known in the art.During for this object, if needed, formula I or formula II compound and/or steric isomer and one or more solids or liquid pharmaceutical excipients and/or assistant agent can be combined, make the suitable administration form or dosage form that can be used as people.
Formula I of the present invention or formula II compound or the pharmaceutical composition containing it can administrations in a unit, and route of administration can be enteron aisle or non-bowel, as oral, muscle, subcutaneous, nasal cavity, oral mucosa, skin, peritonaeum or rectum etc.Form of administration is tablet, capsule, dripping pill, aerosol, pill, pulvis, solution, suspensoid, emulsion, granule, liposome, transdermal agent, buccal tablet, suppository, lyophilized injectable powder etc. such as.Can be ordinary preparation, sustained release preparation, controlled release preparation and various particulate delivery system.In order to unit dosage forms for administration is made tablet, various carrier well known in the art can be widely used.Example about carrier is, such as thinner and absorption agent, as starch, dextrin, calcium sulfate, lactose, N.F,USP MANNITOL, sucrose, sodium-chlor, glucose, urea, calcium carbonate, white bole, Microcrystalline Cellulose, pure aluminium silicate etc.; Wetting agent and tackiness agent, as water, glycerine, polyoxyethylene glycol, ethanol, propyl alcohol, starch slurry, dextrin, syrup, honey, glucose solution, mucialga of arabic gummy, gelatine size, Xylo-Mucine, lac, methylcellulose gum, potassiumphosphate, polyvinylpyrrolidone etc.; Disintegrating agent, such as dry starch, alginates, agar powder, laminaran, sodium bicarbonate and Citric Acid, calcium carbonate, polyoxyethylene, sorbitan fatty acid ester, sodium laurylsulfonate, methylcellulose gum, ethyl cellulose etc.; Disintegration inhibitor, such as sucrose, Tristearoylglycerol, theobroma oil, hydrogenation wet goods; Absorption enhancer, such as quaternary ammonium salt, sodium lauryl sulphate etc.; Lubricant, such as talcum powder, silicon-dioxide, W-Gum, stearate, boric acid, whiteruss, polyoxyethylene glycol etc.Tablet can also be made coating tablet further, such as sugar coated tablet, thin membrane coated tablet, ECT, or double-layer tablets and multilayer tablet.In order to administration unit is made pill, various carrier well known in the art can be widely used.Example about carrier is, such as thinner and absorption agent, as glucose, lactose, starch, theobroma oil, hydrogenated vegetable oil, polyvinylpyrrolidone, Gelucire, kaolin, talcum powder etc.; Tackiness agent is as gum arabic, tragacanth gum, gelatin, ethanol, honey, liquid sugar, rice paste or batter etc.; Disintegrating agent, as agar powder, dry starch, alginates, sodium laurylsulfonate, methylcellulose gum, ethyl cellulose etc.In order to administration unit is made suppository, various carrier well known in the art can be widely used.Example about carrier is, the ester, gelatin, semi-synthetic glyceryl ester etc. of such as polyoxyethylene glycol, Yelkin TTS, theobroma oil, higher alcohols, higher alcohols.In order to administration unit is made capsule, effective constituent formula I or its steric isomer are mixed with above-mentioned various carriers, and the mixture obtained thus is placed in hard obviously capsule or soft capsule.Also effective constituent formula I or its steric isomer can be made microcapsule, be suspended in aqueous medium and form suspensoid, also can load in hard capsule or make injection application.In order to administration unit is made injection preparation, as solution, emulsion, lyophilized injectable powder and suspensoid, all thinners that this area is conventional can be used, such as, the isooctadecanol of water, ethanol, polyoxyethylene glycol, 1,3-PD, ethoxylation, polyoxygenated isooctadecanol, Polyoxyethylene Sorbitol Fatty Acid Esters etc.In addition, in order to prepare isotonic injection liquid, appropriate sodium-chlor, glucose or glycerine can be added in injection preparation, in addition, conventional solubility promoter, buffer reagent, pH adjusting agent etc. can also be added.
In addition, as needs, also tinting material, sanitas, spices, correctives, sweeting agent or other material can be added in pharmaceutical preparation.
Formula I or formula II compound, or the dosage of its pharmacologically acceptable salt depends on many factors, such as, to prevent or the character of disease therapy and severity, the sex of patient or animal, age, body weight and individual reaction, particular compound used, route of administration and administration number of times etc.Above-mentioned dosage can single dose form or be divided into several, such as two, three or four dosage forms for administration.
Term used herein " composition " means to comprise the product of each appointment composition comprising specified amount, and any product directly or indirectly produced from the combination of each appointment composition of specified amount.
The active compound amount of gained the actual dose level of each activeconstituents in pharmaceutical composition of the present invention can be changed, so that effectively can obtain required therapeutic response for concrete patient, composition and administering mode.Dosage level must according to the activity of particular compound, route of administration, treat the severity of the patient's condition and the patient's condition of patient to be treated and medical history and select.But the way of this area is, the dosage of compound, from lower than for obtaining level that required result for the treatment of requires, increases dosage, gradually until obtain required effect.
Another aspect of the invention relates to compound of the present invention or the extract described in any one of the present invention or composition of the present invention purposes in preparation HMG-CoA reductase inhibitor.
Another aspect of the invention relates to method that is in vivo a kind of or vitro inhibition HMG-CoA reductase, comprises the step using the compound of the present invention of significant quantity or the extract described in any one of the present invention or composition of the present invention.
The test of embodiment 4 proves, the suppression HMG-CoA reductase that compound of the present invention has dose-dependently is active.
Another aspect of the invention relates to a kind of reduction or adjusting blood lipid or prevents and/or treats hyperlipemia, hyperlipidemia, hypercholesterolemia or atherosclerotic method, comprises to the step with the extract described in the compound of the present invention or any one of the present invention of significant quantity or composition of the present invention.
When for above-mentioned treat and/or prevent or assisting therapy time, a kind of the compounds of this invention treating and/or preventing significant quantity can be applied in a pure form, or with the acceptable ester of pharmacy or prodrug forms (when there are these forms) application.Or described compound can accept the pharmaceutical composition administration of vehicle containing this object compound and one or more medicines.Term " significant quantity " refers to the dosage that can realize treating, prevent, alleviate and/or alleviating disease of the present invention or illness in experimenter.But it should be understood that total daily dosage portion of the compounds of this invention and composition must be maked decision within the scope of reliable medical judgment by attending physician.For any concrete patient, concrete treatment effective dose level must be determined according to many factors, and described factor comprises treated obstacle and the severity of this obstacle; The activity of the particular compound adopted; The concrete composition adopted; Age of patient, body weight, general health situation, sex and diet; The administration time of the particular compound adopted, route of administration and excretion rate; The treatment time length; The medicine combinationally using with adopted particular compound or use simultaneously; And the known similar factor of medical field.Such as, the way of this area is, the dosage of compound, from lower than for obtaining level that required result for the treatment of requires, increases dosage, gradually until obtain required effect.In general, formula I is used for the dosage of Mammals particularly people can between 0.001-1000mg/kg body weight/day, such as, between 0.01-100mg/kg body weight/day, such as, between 0.01-10mg/kg body weight/day.
Various disease of the present invention or illness effectively can be prevented and/or treated according to compound of the present invention.
In the present invention, term " C
1-C
3alkyl " comprise methyl, ethyl, propyl group or sec.-propyl.
The beneficial effect of the invention
Compound of the present invention can suppress HMG-CoA reductase effectively, and the suppression HMG-CoA reductase with dose-dependently is active; Have as reducing or adjusting blood lipid or prevent and/or treat the potentiality of hyperlipemia, hyperlipidemia, hypercholesterolemia or atherosclerotic medicine.
Embodiment
Below in conjunction with embodiment, embodiment of the present invention are described in detail, but it will be understood to those of skill in the art that the following example only for illustration of the present invention, and should not be considered as limiting scope of the present invention.Unreceipted actual conditions person in embodiment, the condition of conveniently conditioned disjunction manufacturers suggestion is carried out.Agents useful for same or the unreceipted production firm person of instrument, being can by the conventional products of commercial acquisition.
embodiment 1: the preparation (1) of formula II compound
Operation steps:
1) get XUEZHIKANG JIAONANG (Beijing University's dimension letter is produced) content dry powder and be about 1kg, be that solvent supersonic extracts 3 times with the methylene dichloride of 2-6 times of volume, each 20-40 minute, united extraction liquid, concentrating under reduced pressure recycling design, obtain methylene dichloride extract 91g.
2) get methylene dichloride extract 50g, upper silica gel column chromatography is separated, and carries out gradient elution by sherwood oil and ethyl acetate.The volume ratio of petroleum ether-ethyl acetate is followed successively by 75: 25, and 50: 50,25: 75,0: 100.
3) getting petroleum ether-ethyl acetate is 25: 75 part 5.0g, be separated through C18 reversed-phase column chromatography, methanol-water (10: 90-100: 0) gradient elution obtains 4 part (methanol-waters 10: 90, 50: 50, 75: 25, 100: 0), wherein methanol-water (75: 25) elution fraction 1.3g half preparative high-performance liquid chromatographic is purified, with acetonitrile-0.2% acetic acid aqueous solution (45: 55) for moving phase, flow velocity is 4mL/min, C18 half preparative chromatography post (10 × 250mm, 5 μm) be stationary phase, DAD detector determined wavelength is 270nm, collect the chromatographic peak of 9.2min, repeatedly cumulative rear concentrated, lyophilize must be about 40mg by this compound.
embodiment 2: the preparation (2) of formula II compound
Operation steps:
1) getting red colouring agent for food, also used as a Chinese medicine 5kg, is that solvent supersonic extracts 3 times with the methylene dichloride of 2-6 times of volume, each 20-40 minute, united extraction liquid, concentrating under reduced pressure recycling design, obtains methylene dichloride extract 78g.
2) get methylene dichloride extract 30g, upper silica gel column chromatography is separated, and carries out gradient elution by sherwood oil and ethyl acetate.The volume ratio of petroleum ether-ethyl acetate is followed successively by 75: 25, and 50: 50,25: 75,0: 100.
3) getting petroleum ether-ethyl acetate is 25: 75 part 3.0g, be separated through C18 reversed-phase column chromatography, methanol-water (10: 90-100: 0) gradient elution obtains 4 part (methanol-waters 10: 90, 50: 50, 75: 25, 100: 0), wherein methanol-water (75: 25) elution fraction 0.8g half preparative high-performance liquid chromatographic is purified, with acetonitrile-0.2% acetic acid aqueous solution (45: 55) for moving phase, flow velocity is 4mL/min, C18 half preparative chromatography post (10 × 250mm, 5 μm) be stationary phase, DAD detector determined wavelength is 270nm, collect the chromatographic peak of 9.2min, repeatedly cumulative rear concentrated, lyophilize must be about 25mg by this compound.
embodiment 3: the Structural Identification of compound
Specimen in use is compound prepared by embodiment 1 and 2.
1. the physicochemical data of compound
White powder, specific rotation: [α]
25 d-50.00 (c0.118, CH
2cl
2: MeOH=1: 1);
There are three maximum absorption bands in UV spectrum, are respectively λ
max(CH
2cl
2: MeOH)=271.6nm, 282.2nm, 293.8nm.
FT-IR (KBr, cm
-1) spectrum: 3392 (-OH), 2969,2933 (saturated hydrocarbon), 1652,1647 (C=C), 1456,1378 (gem-dimethyls).
2. the determination of molecular formula
The m/z483.3104 [M+Na] that HR-ESI-MS provides
+(calcd.483.3081, err2.3), the molecular weight being inferred as this compound is 460.32.?
1h-NMR and
13c-NMR shows, and has 40 hydrogen signals and 28 carbon signals.From DEPT display, there are 6 quaternary carbons, 10 CH, 6 CH
2and 6 CH
3.?
13in C-NMR, there are 4 olefinic carbon signals at 117.2ppm, 119.6ppm, 139.1ppm and 140.6ppm place.With HSQC scheme with
13c-NMR figure binding analysis, there are 6 carbon be connected with Sauerstoffatom in 70.5ppm, 72.4ppm, 74.5ppm, 80.4ppm, 83.8ppm and 84.4ppm place.Binding molecule amount and
1h,
13c-NMR and DEPT signal graph, if this compound has 6 Sauerstoffatom molecular weight more than 460.32, infers that this compound has 5 Sauerstoffatoms and do not embody 4 hydrogen atoms of hydrogen spectrum signal thus.Thus can judge that, in above-mentioned 5 Sauerstoffatoms, 4 Sauerstoffatoms are attributed to 4 hydroxyls, other 5th Sauerstoffatom exists with the form of ether.According to above-mentioned analysis, can determine there are 28 carbon atoms in this molecule, 44 hydrogen atoms and 5 Sauerstoffatoms, molecular formula is C
28h
44o
5.
3. the determination of structural formula
The carbon analyzing this compound is composed (
13c-NMR and DEPT), have 28 carbon atoms, wherein 6 carbon are methyl.Based on 3 maximum absorption at 271.6nm, 282.2nm and 293.8nm place in ultra-violet absorption spectrum and ergosterol class substantially identical, tentatively infer the skeleton that this compound has ergosterol.Can calculate its degree of unsaturation from molecular formula is 7.Thus can infer: this compound, except 6 unsaturated places of 2 double bonds and sterol backbone 4 rings, also has a unsaturated place and can only be a ring herein, can judge that this ring is by the oxirane ring formed centered by the 5th Sauerstoffatom thus.HSQC figure combines with HMBC coherent signal figure and analyzes, and 6 carbon be connected with Sauerstoffatom are attributed to 4 carbon (70.5 that 4 hydroxyls are connected respectively, 72.4,74.5,80.4ppm) and 2 carbon (83.8,84.4ppm) being connected with surplus next Sauerstoffatom.Deep HMBC map analysis, can infer that the 5th member ring systems is by C-16 (83.8ppm), C-17 (66.9ppm), C-20 (80.4ppm), 5 rings of C-22 (84.4ppm) and Sauerstoffatom composition.This not only meets degree of unsaturation, but also meets the carbonatoms be connected with Sauerstoffatom.The molecular weight that further analysis mass spectrometry system provides and molecular formula, confirm above-mentioned analysis.The above analysis can infer that this new compound is: 16,22-epoxy ergot steroid-5,7-diene-3,20,23,25-tetrol
I.e. (16,22-epoxy-ergosta-5,7-dien-3,20,23,25-tetraol).Structural formula is for shown in formula II below.
4. the NMR data of new compound
As shown in Table 1 below.
Table 1: NMR data (600MHz, the CDCl of new compound
3, JinHz)
Note: "-" represents to there is not coherent signal.
embodiment 4:HMG-CoA reductase active is tested
1. experiment material
1.1 medicine
New compound---prepared by embodiment 1 or 2.
Lovastatin standard substance---be purchased from Sigma.
1.2 enzyme
Rat liver microsome (HMG-CoA reductase)----, can be purchased, also can with reference to following preparation method: the liver taking out male rat, after KESD wash buffer, the centrifugal 15min of 1200g, gets supernatant liquor.After using the centrifugal 90min of 105,000g twice again, collect centrifugation.8.3% glycerine is added, with 37 DEG C of temperature bath heating 1h in centrifugation.Rat liver microsome runic thing saturated ammonium sulphate purifying also collects 35-50% purification part.The purification part obtained can be left in-80 DEG C of refrigerators.
1.3 reagent
Repone K, potassium primary phosphate, ethylenediamine tetraacetic acid (EDTA), dithiothreitol (DTT)---be purchased from Beijing chemical reagents corporation;
Reduced nicotinamide-adenine dinucleotide (NADPH)---be purchased from Merk;
3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA)----be purchased from Sigma.
2. experimental technique
New compound is used 75% dissolve with ethanol solution, initial concentration is 8.0mg/mL and stepwise dilution, obtains 4.0mg/mL, 2.0mg/mL, 1.0mg/mL; Lovastatin is positive control, uses 75% dissolve with ethanol solution, and concentration is 2.0mg/mL; In mensuration system, cumulative volume is 250 μ L, the concentration of each composition is: Repone K 200mM, potassium primary phosphate 160mM, ethylenediamine tetraacetic acid (EDTA) 4mM, dithiothreitol (DTT) 10mM, the concentration of two substrate Reduced nicotinamide-adenine dinucleotides and 3-hydroxy-3-methylglutaryl-coenzyme A is respectively 200 μMs and 50 μMs, pH6.8, enzyme adds 30 μ L, 4 test group respectively add the new compound solution of 10 μ L different concns, positive controls adds 10 μ L lovastatin solution, and blank group adds 10 μ L75% ethanolic solns, and Versamax microplate reader detects OD under 37 DEG C of conditions
340dynamic change.By detecting OD
340the speed (representing with slope value) declined in 5 minutes, evaluates the power of HMG-CoA reductase activity, and then evaluates the power of inhibitor activity, the results are shown in Table 2.
3. experimental result
As shown in Table 2 below.
Table 2: inhibitor activity detected result
* blank is solvent.
Lovastatin is positive control.
Experimental result shows, the activity of new compound to HMG-CoA reductase has restraining effect, and in concentration-effect relation.Its IC
50value is about 250 μ g/mL, shows that the activity of this compound to HMG-CoA reductase has good restraining effect.
Although the specific embodiment of the present invention has obtained detailed description, it will be understood to those of skill in the art that.According to disclosed all instructions, can carry out various amendment and replacement to those details, these change all within protection scope of the present invention.Four corner of the present invention is provided by claims and any equivalent thereof.
Claims (14)
1. the compound shown in formula I, or its pharmacy acceptable salt,
Wherein,
R
1for-OH;
R
2for-OH;
R
3be selected from-OH, H and C
1-C
3alkyl;
R
4be selected from-OH, H and C
1-C
3alkyl.
2. compound according to claim 1 or its pharmacy acceptable salt, it is the compound shown in formula II below, or its pharmacy acceptable salt,
3. the preparation method of compound according to claim 2, comprises the steps:
1) alcohol extract of red colouring agent for food, also used as a Chinese medicine and/or red colouring agent for food, also used as a Chinese medicine is got, one or many is extracted as solvent supersonic with one or more organic solvents be selected from methylene dichloride, ethyl acetate, acetone, methyl alcohol, ethanol of 2-6 times of volume, each 20-40 minute, united extraction liquid, except desolventizing, obtain extract;
2) by step 1) extract that obtains carries out silica gel column chromatography separation, carries out gradient elution by sherwood oil and ethyl acetate; The volume ratio of petroleum ether-ethyl acetate is followed successively by 75:25,50:50-25:75,0:100;
3) get step 2) in petroleum ether-ethyl acetate be the elution fraction of 50:50-25:75, be separated through C18 reversed-phase column chromatography, carry out gradient elution with methanol-water, the volume ratio of methanol-water is followed successively by 10:90,50:50-75:25,100:0;
4) get step 3) in methanol-water be that the elution fraction of 50:50-75:25 is purified with half preparative high-performance liquid chromatographic, with acetonitrile-0.2% acetic acid aqueous solution of 45:55 for moving phase, C18 semipreparative column is stationary phase, collects the chromatographic peak part of 9.2min; With
5) by step 4) product carry out lyophilize, obtain formula II compound.
4. preparation method according to claim 3, wherein, the alcohol extract of red colouring agent for food, also used as a Chinese medicine is XUEZHIKANG JIAONANG content.
5. the preparation method according to claim 3 or 4, it meets any one in following (1)-(9) or multinomial:
(1) step 1) in, described organic solvent is methylene dichloride;
(2) step 1) in, described supersound extraction carries out 3 times;
(3) step 1) in, by concentrating under reduced pressure except desolventizing;
(4) step 2) in, the volume ratio of petroleum ether-ethyl acetate is followed successively by 75:25,50:50,25:75,0:100;
(5) step 3) in, get step 2) in petroleum ether-ethyl acetate be the elution fraction of 50:50 or 25:75;
(6) step 3) in, the volume ratio of methanol-water is followed successively by 10:90,50:50,75:25,100:0;
(7) step 4) in, get step 3) in methanol-water be the elution fraction of 50:50 or 75:25;
(8) step 4) in, the chromatographic peak part of the 9.2min collected is merged; With
(9) step 5) in, cryodesiccated condition is: condenser temperature-40 to-85 DEG C, vacuum tightness 0-100Pa.
6. preparation method according to claim 5, wherein, condenser temperature-50 to-82.7 DEG C, vacuum tightness 2-13Pa.
7. preparation method according to claim 5, wherein, condenser temperature-82.7 DEG C, vacuum tightness 2Pa, or condenser temperature-50 DEG C, vacuum tightness 8.5Pa.
8. an extract, it contains formula II compound according to claim 2, and its be following (1) to any one in (3):
(1) step 1 in claim 3) to 2) in obtained petroleum ether-ethyl acetate be the elution fraction of 50:50-25:75;
(2) step 1 in claim 3) to 3) in obtained methanol-water be the elution fraction of 50:50-75:25; With
(3) step 1 in claim 3) to 4) in the chromatographic peak part of obtained 9.2min.
9. extract according to claim 8, it is the extract of XUEZHIKANG JIAONANG content dry powder or the extract of red colouring agent for food, also used as a Chinese medicine.
10. a composition, it is made up of the extract described in claim 8 or 9 and pharmaceutically acceptable carrier or auxiliary material.
11. 1 kinds of compositions, it is made up of the compound described in claim 1 or 2 and pharmaceutically acceptable carrier or auxiliary material.
Compound described in 12. claims 1 or 2 or the extract described in claim 8 or 9 or the composition described in claim 10 or 11 reduce or adjusting blood lipid or the purposes that prevents and/or treats in hyperlipemia, hyperlipidemia, hypercholesterolemia or atherosclerotic medicine in preparation.
Compound described in 13. claims 1 or 2 or the extract described in claim 8 or 9 or the purposes of the composition described in claim 10 or 11 in preparation HMG-CoA reductase inhibitor.
14. 1 kinds of methods suppressing HMG-CoA reductase in vitro, comprise the step using the compound described in claim 1 or 2 of significant quantity or the extract described in claim 8 or 9 or the composition described in claim 10 or 11.
Priority Applications (15)
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| CN201110441993.2A CN103172693B (en) | 2011-12-26 | 2011-12-26 | A kind of sterol derivative, Preparation Method And The Use |
| CN201280062316.2A CN104024270B (en) | 2011-12-26 | 2012-12-25 | A kind of sterol derivative, Preparation Method And The Use |
| SG11201403618PA SG11201403618PA (en) | 2011-12-26 | 2012-12-25 | A sterol derivative and preparation method and uses thereof |
| MYPI2014001905A MY172011A (en) | 2011-12-26 | 2012-12-25 | Sterols derivative, and preparation method and purpose thereof |
| EP12862540.7A EP2799444B1 (en) | 2011-12-26 | 2012-12-25 | Sterols derivative, and preparation method and purpose thereof |
| TW101149915A TWI518094B (en) | 2011-12-26 | 2012-12-25 | One kind of derivatives of sterols, their preparation and use |
| PCT/CN2012/087360 WO2013097681A1 (en) | 2011-12-26 | 2012-12-25 | Sterols derivative, and preparation method and purpose thereof |
| US14/368,494 US10889612B2 (en) | 2011-12-26 | 2012-12-25 | Sterol derivatives and preparation method and uses thereof |
| ES12862540.7T ES2666459T3 (en) | 2011-12-26 | 2012-12-25 | Sterol derivative, preparation procedure and purpose thereof |
| HK14112046.3A HK1198539A1 (en) | 2011-12-26 | 2014-11-28 | Sterols derivative, and preparation method and purpose thereof |
| US17/071,958 US20210024570A1 (en) | 2011-12-26 | 2020-10-15 | Sterol derivatives and preparation method and uses thereof |
| US17/071,963 US11634454B2 (en) | 2011-12-26 | 2020-10-15 | Sterol derivatives and preparation method and uses thereof |
| US17/818,302 US11845774B2 (en) | 2011-12-26 | 2022-08-08 | Sterol derivatives and preparation method and uses thereof |
| US17/818,667 US11845775B2 (en) | 2011-12-26 | 2022-08-09 | Sterol derivatives and preparation method and uses thereof |
| US17/820,553 US20220402967A1 (en) | 2011-12-26 | 2022-08-17 | Sterol derivatives and preparation method and uses thereof |
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| CN101104003A (en) * | 2006-07-11 | 2008-01-16 | 北京北大维信生物科技有限公司 | Xuezhikang purpose-made monascus anti-cancer new use |
| CN101313919A (en) * | 2007-05-31 | 2008-12-03 | 北京北大维信生物科技有限公司 | Rice fermented with red yeast extract, preparation method and quality detection method thereof |
| CN101469014A (en) * | 2007-12-27 | 2009-07-01 | 北京北大维信生物科技有限公司 | Novel compound and separation method thereof |
| CN101942003A (en) * | 2009-07-09 | 2011-01-12 | 北京北大维信生物科技有限公司 | Compound and preparation method thereof |
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| CN101104003A (en) * | 2006-07-11 | 2008-01-16 | 北京北大维信生物科技有限公司 | Xuezhikang purpose-made monascus anti-cancer new use |
| CN101313919A (en) * | 2007-05-31 | 2008-12-03 | 北京北大维信生物科技有限公司 | Rice fermented with red yeast extract, preparation method and quality detection method thereof |
| CN101469014A (en) * | 2007-12-27 | 2009-07-01 | 北京北大维信生物科技有限公司 | Novel compound and separation method thereof |
| CN101942003A (en) * | 2009-07-09 | 2011-01-12 | 北京北大维信生物科技有限公司 | Compound and preparation method thereof |
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