CN103880678B - Benzoic acid derivative and preparation thereof and hypoglycemic application - Google Patents
Benzoic acid derivative and preparation thereof and hypoglycemic application Download PDFInfo
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Abstract
The invention discloses benzoic acid derivative and preparation method thereof and application.Benzoic acid derivative provided by the present invention is for such as formula the compound shown in I or formula II; In described formula I, R
1and R
2for following a)-d) in arbitrary: a) R
1=CH
3, R
2=COOH; B) R
1=CH
3, R
2=COOCH
3; C) R
1=CH
3, R
2=CHO; D) R
1=CH
2oH, R
2=CH
2oCH
3; In described formula II, R
1for following e)-g) in arbitrary: e) R
1=CH
2cH
2cH
2cOOH; F) R
1=CH (COOH) CH (CH
3) CH
2cH
3; G) R
1=CH (COOH) CH
2cH (CH
3) CH
3.Experiment proves, the compounds of this invention has the activity of very strong Inhibiting α-glucosidase, and obviously can reduce the blood glucose value of type II diabetes rat model.Compound provided by the present invention and pharmacologically acceptable salt thereof are that exploitation treats and/or prevents the original new drug of type ii diabetes, the innovative ofhypoglycemic medicine of exploitation Inhibiting α-glucosidase, and food, functional health care product etc. that exploitation has an above-mentioned functions provide new basic substance, have potential huge Social benefit and economic benefit.
formula I
Description
Technical field
The present invention relates to benzoic acid derivative and preparation method thereof and application, particularly two classes totally 7 kinds of benzoic acid derivatives, preparation methods, and as the application in alpha-glucosidase inhibitor.
Background technology
Type ii diabetes has become in world wide the Non Communicable Diseases (NCD) having a strong impact on human health, has more than 2,000 ten thousand China diabetic subject, and wherein nearly 90% is type ii diabetes, and occurs feature that is multiple, rejuvenation, serious threat national health.The Regular Insulin of type ii diabetes patient is in relative deficiency state, therefore its treatment can rely on Regular Insulin completely and use other medicines.The main method of current treatment diabetes is the hypoglycemic drug such as insulinize and oral sulfourea, biguanides, but insulinize is expensive, and other types of drug exists the features such as offer limited effectiveness, toxic side effect be obvious.
Alpha-glucosidase acts on the oligosaccharides such as enteral sucrose, and final generation glucose also enters blood through intestinal absorption, is the major cause that postprandial blood sugar raises.Postprandial hyperglycemia can cause body reduce the susceptibility of Regular Insulin thus add type ii diabetes conditions of patients, even causes a series of complication.Thus the control that will significantly improve diabetes of alpha-glucosidase inhibitor.The alpha-glucosidase inhibitor used in the market is mainly acarbose, voglibose, miglitol etc., but it should be noted that, these inhibitor of current use all bring toxic side effect in varying degrees, as gastrointestinal side effect, urticaria, hepatic insufficiency, cardiac system risk etc.Therefore, the alpha-glucosidase inhibitor that develop actively is safer, more potent is necessary and urgent.
Summary of the invention
An object of the present invention is to provide two classes totally 7 kinds of benzoic acid derivatives.
Benzoic acid derivative provided by the present invention, for such as formula the compound shown in I and formula II:
R
1for CH
3; R
2during for COOH, compound shown in formula I is such as formula shown in I-1.
R
1for CH
3; R
2for COOCH
3time, compound shown in formula I is such as formula shown in I-2.
R
1for CH
3; R
2during for CHO, compound shown in formula I is such as formula shown in I-3.
R
1for CH
2oH; R
2for CH
2oCH
3time, compound shown in formula I is such as formula shown in I-4.
R
1for CH
2cH
2cH
2during COOH, compound shown in formula II is such as formula shown in II-1.
R
1for CH (COOH) CH (CH
3) CH
2cH
3time, compound shown in formula II is such as formula shown in II-2.
R
1for CH (COOH) CH
2cH (CH
3) CH
3time, compound shown in formula II is such as formula shown in II-3.
In above-mentioned 7 compounds, Compound nomenclature shown in formula I-1 is sterenolA, Compound nomenclature shown in formula I-2 is sterenolB, Compound nomenclature shown in formula I-3 is sterenolC, Compound nomenclature shown in formula I-4 is sterenolD, Compound nomenclature shown in formula II-1 is stereninE, and Compound nomenclature shown in formula II-2 is stereninF, and Compound nomenclature shown in formula II-3 is stereninG.
The application of described compound or pharmaceutically acceptable salt thereof in following (A1)-(A4) is arbitrary also belongs to protection scope of the present invention:
(A1) as alpha-glucosidase inhibitor;
(A2) preparation treats and/or prevents the medicine of type ii diabetes;
(A3) medicine of Inhibiting α-glucosidase is prepared;
(A4) preparation has food or the functional health care product of blood sugar reducing function.
Another object of the present invention is to provide a kind of medicine treating and/or preventing type ii diabetes.
The medicine treating and/or preventing type ii diabetes provided by the present invention, its activeconstituents is described compound or pharmaceutically acceptable salt thereof.
Also object of the present invention is to provide a kind of medicine of Inhibiting α-glucosidase.
The medicine of Inhibiting α-glucosidase provided by the present invention, its activeconstituents is described compound or pharmaceutically acceptable salt thereof.
Another object of the present invention is to provide a kind of food or the functional health care product with blood sugar reducing function.
Food or the functional health care product with blood sugar reducing function provided by the present invention, its activeconstituents is described compound or pharmaceutically acceptable salt thereof.
Described medicines all above all can be tablet, pulvis, capsule, oral liquid, emulsion, paste, creme, injection, suspensoid, tincture, granule or aerosol.The medicine of above-mentioned various formulation all can be prepared according to the ordinary method of pharmaceutical field.
In the present invention, above all described in treat and/or prevent type ii diabetes and be embodied in: the fasting blood sugar reducing type II diabetes rat model.
Another object of the present invention is to provide the preparation method of described compound.
The preparation method of described compound, specifically can comprise the steps:
(1) thick hair Boreostereum vibrans (Stereumhirsutum) is inoculated on solid medium carries out solid fermentation cultivation;
(2) in the system after step (1) fermentation, add the extracting solution that organic solvent soak extraction obtains containing compound shown in compound and formula II shown in formula I, by described extracting solution in 36-40 DEG C of (as 38 DEG C) concentrating under reduced pressure, vacuum-drying (vacuum tightness <3mmHg), obtains the crude extract containing compound shown in compound and formula II shown in formula I.
In step (1), described solid medium can be following a1) or a2):
A1) substratum be made up of 1 weight part base material and 1-1.5 weight parts water;
A2) be made up of 1 weight part base material, 1-1.5 weight parts water and 0.02-0.1 weight part sugar;
Described base material can be at least one in long-grained nonglutinous rice, wheat, Gorgon fruit, oat, polished rice, corn, glutinous rice and the seed of Job's tears; Described sugar can be glucose.
Specifically, described solid medium can be following substratum first, substratum second or substratum third.Described substratum first is made up of long-grained nonglutinous rice and water, and proportioning is 80g long-grained nonglutinous rice: 100ml water.Described substratum second is made up of wheat and water, and proportioning is 80g wheat: 90ml water.Described substratum third is made up of Gorgon fruit, glucose and water, and proportioning is 80g Gorgon fruit: 5g glucose: 90ml water.
In step (1), the temperature that described solid fermentation is cultivated can be 22-28 DEG C, and as 25 DEG C, mode is lucifuge quiescent culture, and the time was 10-50 days, as 40 days.
In step (2), described organic solvent specifically can be ethyl acetate.
In step (2), described soak extraction obtains the method for the extracting solution containing compound shown in compound and formula II shown in formula I, specifically can comprise following b1)-b4) step:
B1) first time soak extraction: add ethyl acetate and soak 7 days in the system after step (1) fermentation, period every 8h supersound process once, gets supernatant liquor after immersion;
B2) second time soak extraction: to step b1) residuum in add ethyl acetate and soak 7 days, period every 8h supersound process once, gets supernatant liquor after immersion;
B3) third time soak extraction: to step b2) residuum in add ethyl acetate and soak 7 days, period every 8h supersound process once, gets supernatant liquor after immersion;
B4) combining step b1)-b3) supernatant liquor, namely obtain the described extracting solution containing compound shown in compound and formula II shown in formula I.
At b1)-b3) in, be once describedly ultrasonicly 50Hz continuous ultrasound 20min.In the present invention, temperature when soaking for three times is all specially 20 DEG C.
Further, described method also comprises the steps (3)-(5):
(3) the described crude extract containing compound shown in compound with formula II shown in formula I step (2) obtained carries out silica gel column chromatography and is separated, using the mixed solution of chloroform and acetone as moving phase, the gradient being followed successively by 100:0,100:1,50:1,30:1,20:1,10:1,8:2,7:3,6:4 and 0:1 according to the volume ratio of chloroform in moving phase and acetone carries out gradient elution (during wash-out, flow velocity specifically can be 10ml per minute), and the elution volume of the moving phase of each gradient is 1500 milliliters; The volume ratio of collecting chloroform and acetone is the elutriant of first 500 milliliters of the moving phase of 50:1, is designated as solution first; The volume ratio of collecting chloroform and acetone is the elutriant of latter 500 milliliters of the moving phase of 20:1, and the volume ratio of chloroform and acetone is the elutriant of first 500 milliliters of the moving phase of 10:1, and after mixing, solution is designated as solution second.
Wherein, described crude extract is carried out silica gel column chromatography be separated into: be that the chloroform of 1:1 and the mixed solution of methyl alcohol dissolve (wherein by described crude extract volume ratio, described crude extract and described volume ratio are that the proportioning of the chloroform of 1:1 and the mixed solution of methyl alcohol is as 13g:50ml), centrifugal (5min as centrifugal in 5000rpm), get supernatant liquor, carry out described silica gel column chromatography.When carrying out described silica gel column chromatography, the silicagel column filler of employing is 200-300 object silica gel particle, and silicagel column specification is Φ 4.5 × 80cm, and amount of filler is 213g.
(4) the solution first that step (3) obtains is carried out gel permeation chromatography, carry out wash-out (during wash-out, flow velocity specifically can be 1ml per minute) using methyl alcohol as moving phase; Collect the elutriant of the 140th milliliter-180 milliliters, in 36-40 DEG C of (as 38 DEG C) concentrating under reduced pressure, vacuum-drying (vacuum tightness <3mmHg), obtains compound shown in formula I-1; Collect the elutriant of the 200th milliliter-240 milliliters, in 36-40 DEG C of (as 38 DEG C) concentrating under reduced pressure, vacuum-drying (vacuum tightness <3mmHg), obtains compound shown in formula I-2; Collect the elutriant of the 260th milliliter-340 milliliters, in 36-40 DEG C of (as 38 DEG C) concentrating under reduced pressure, vacuum-drying (vacuum tightness <3mmHg), obtains compound shown in formula I-3.
Wherein, described solution first is carried out gel permeation chromatography is: by described solution first in 36-40 DEG C of (as 38 DEG C) concentrating under reduced pressure, vacuum-drying (vacuum tightness <3mmHg), obtain dry-matter first, with dry-matter first described in dissolve with methanol (wherein, the proportioning of described dry-matter first and described methyl alcohol is as 2g:3ml), centrifugal (the centrifugal 5min of 5000rpm), gets supernatant liquor and carries out described gel permeation chromatography.When carrying out described gel permeation chromatography, the gel column filler of employing is sephadexLH-20, and gel column specification is Φ 2.5 × 120cm.
(5) the solution second that step (3) obtains is carried out ODS reversed-phase silica gel chromatography, using the mixed solution of first alcohol and water as moving phase, the gradient being followed successively by 20:80,30:70,40:60,50:50,60:40,85:15 and 0:1 according to the volume ratio of first alcohol and water in moving phase carries out gradient elution (flow velocity during wash-out specifically can be 3ml per minute), and the elution volume of the moving phase of each gradient is 200 milliliters; The volume ratio of collecting first alcohol and water is the elutriant of latter 100 milliliters of the moving phase of 40:60, and in 36-40 DEG C of (as 38 DEG C) concentrating under reduced pressure, vacuum-drying (vacuum tightness <3mmHg), obtains compound shown in formula I-4; The volume ratio of collecting first alcohol and water is the elutriant of latter 100 milliliters of the moving phase of 50:50, and in 36-40 DEG C of (as 38 DEG C) concentrating under reduced pressure, vacuum-drying (vacuum tightness <3mmHg), obtains compound shown in formula II-1; The volume ratio of collecting first alcohol and water is the elutriant of latter 100 milliliters of the moving phase of 60:40, and in 36-40 DEG C of (as 38 DEG C) concentrating under reduced pressure, vacuum-drying (vacuum tightness <3mmHg), obtains compound shown in formula II-2; The volume ratio of collecting first alcohol and water is the elutriant of latter 100 milliliters of the moving phase of 85:15, and in 36-40 DEG C of (as 38 DEG C) concentrating under reduced pressure, vacuum-drying (vacuum tightness <3mmHg), obtains compound shown in formula II-3.
Wherein, described solution second is carried out ODS reversed-phase silica gel chromatography is: by described solution second in 36-40 DEG C of (as 38 DEG C) concentrating under reduced pressure, vacuum-drying (vacuum tightness <3mmHg), obtain dry-matter fourth, with dry-matter fourth described in dissolve with methanol (wherein, the proportioning of described dry-matter fourth and described methyl alcohol is 390.5mg:1.5ml), centrifugal (the centrifugal 5min of 5000rpm), gets supernatant liquor and carries out described ODS reversed-phase silica gel chromatography.When carrying out described ODS reversed-phase silica gel chromatography, the silicagel column filler of employing is ODS reverse phase silica gel (specification brand is specially " YMC*GEL, 12nm, S-50 μm "), and the mesolow chromatographic column of silicagel column to be specification be Φ 3.0 × 30cm, amount of filler is 40.4g.
In the step (1) of described method, before thick hair Boreostereum vibrans (Stereumhirsutum) being inoculated into solid medium carries out solid fermentation, also comprise the steps: described thick hair Boreostereum vibrans (Stereumhirsutum) to be inoculated on PDA substratum to carry out activation culture, get the mycelium after activation culture and be transferred on liquid PDB substratum and carry out seed culture.The condition of described activation culture is that 25 DEG C of lucifuges cultivate 7 days; The condition of described seed culture is 28 DEG C, 180rmp lucifuge cultivates 7 days.Consisting of of described PDA substratum: potato 200g, glucose 20g, agar powder 18g, be settled to 1L with distilled water; Consisting of of described liquid PDB substratum: potato 200g, glucose 20g, distilled water is settled to 1L.
In the process, described thick hair Boreostereum vibrans (Stereumhirsutum) specifically can be thick hair Boreostereum vibrans (Stereumhirsutum) THG20.
Experiment proves, shown in formula I, compounds exhibit shown in compound and formula II goes out the activity of very strong Inhibiting α-glucosidase.By measuring two compounds, alpha-glucosidase half-inhibition concentration (IC50 value) is shown: the intensity of the Inhibiting α-glucosidase of compound shown in formula I-1 is 84 times of positive control medicine acarbose; The intensity of the Inhibiting α-glucosidase of compound shown in formula I-2 is 209 times of positive control medicine acarbose; The intensity of the Inhibiting α-glucosidase of compound shown in formula I-3 is 106 times of positive control medicine acarbose; The intensity of the Inhibiting α-glucosidase of compound shown in formula I-4 is 28 times of positive control medicine acarbose; The intensity of the Inhibiting α-glucosidase of compound shown in formula II-1 is 17 times of positive control medicine acarbose, and the intensity of the Inhibiting α-glucosidase of compound shown in formula II-2 is 49 times of positive control medicine acarbose; The intensity of the Inhibiting α-glucosidase of compound shown in formula II-3 is 23 times of positive control medicine acarbose.In addition, in experimentation on animals, 7 compounds all obviously can reduce the blood glucose value of type II diabetes rat model.The present invention is that exploitation treats and/or prevents the original new drug of type ii diabetes, the innovative ofhypoglycemic medicine of exploitation Inhibiting α-glucosidase, and food, functional health care product etc. that exploitation has an above-mentioned functions provide new basic substance, have potential huge Social benefit and economic benefit.
Embodiment
The experimental technique used in following embodiment if no special instructions, is ordinary method.
Material used in following embodiment, reagent etc., if no special instructions, all can obtain from commercial channels.
Thick hair Boreostereum vibrans (Stereumhirsutum) THG20: (be called for short CGMCC at China Committee for Culture Collection of Microorganisms's common micro-organisms center, address is: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica) deposit number be CGMCC5.1975.The public can buy from CGMCC and obtain.
Alpha-glucosidase: buy from Sigma company (article No. is G5003); 4-oil of mirbane-α-D-glucopyranoside: buy from lark prestige Science and Technology Ltd. (article No. is 270305); U-9889: buy from lark prestige Science and Technology Ltd. (article No. is M02540).Wistar male rat (SPF level): Beijing Vital River Experimental Animals Technology Co., Ltd. provides.
The preparation of compound shown in compound and formula II shown in embodiment 1, formula I
One, bacterial strain activation
Thick hair Boreostereum vibrans (Stereumhirsutum) THG20 is seeded to PDA culture medium flat plate, and 25 DEG C of lucifuges are cultivated 7 days (mycelia covers with whole flat board).
PDA substratum (natural pH): potato 200g, glucose 20g, agar powder 18g, be settled to 1L with distilled water.
Two, seed culture
Mycelium in step one is forwarded to liquid PDB substratum, 28 DEG C, 180rmp lucifuge cultivates 7 days, obtains seed culture fluid.
PDB substratum (natural pH): potato 200g, glucose 20g, distilled water is settled to 1L.
Three, solid fermentation is cultivated
Get the seed culture fluid that 5ml step 2 obtains, be seeded to solid medium and (in each 500ml triangular flask, a solid medium be housed, 10 triangular flasks are set, every part of solid medium is made up of 80g long-grained nonglutinous rice and 100ml water), 25 DEG C of darkroom quiescent culture 40 days, obtain fermented product (the whole culture system that the material produced after namely being utilized by bacterium by the metabolite of thalline, bacterium, substratum and substratum forms).
Wherein, the composition of solid medium as above available following (a1) or (a2) substitutes: (a1) 80g wheat and 90ml water; (a2) 80g Gorgon fruit, 5g glucose and 90ml water.
Four, the preparation of compound
Get the fermented product that step 3 obtains, add 300 milliliters of ethyl acetate room temperatures (20 DEG C) lixiviate 7 days (period carried out a supersound process every 8 hours, and each supersound process is 50Hz continuous ultrasound 20min) in each triangular flask, get supernatant liquor; Residuum adds 300 milliliters of new ethyl acetate normal temperature (20 DEG C) lixiviate 7 days (period carried out a supersound process every 8 hours, and each supersound process is 50Hz continuous ultrasound 20min) again, gets supernatant liquor; Residuum adds 300 milliliters of new ethyl acetate normal temperature (20 DEG C) lixiviate 7 days (period carried out a supersound process every 8 hours, and each supersound process is 50Hz continuous ultrasound 20min), gets supernatant liquor; The supernatant liquor three lixiviates obtained merges, and then in 38 DEG C of concentrating under reduced pressure, vacuum tightness <3mmHg vacuum-drying, obtains 13.0g medicinal extract.
By above-mentioned 13.0g medicinal extract, dissolve with 50ml chloroform-methanol (volume ratio of chloroform and methyl alcohol is 1:1), the centrifugal 5min of 5000rpm, collect supernatant liquor, by supernatant liquor and 13.0g column chromatography silica gel (Qingdao Marine Chemical Co., Ltd., size is 200-300 order) mixing, evaporate to dryness organic solvent in 50 DEG C of water-baths, then the open column chromatography post of silica gel (Φ 4.5 × 80cm is added into, be filled with 200g column chromatography silica gel, the particle diameter that Qingdao Marine Chemical Co., Ltd. produces is 200-300 object column chromatography silica gel), chloroform-acetone is adopted to carry out gradient elution (in gradient elution process as moving phase again, volume ratio is adopted to be 100:0 successively, 100:1, 50:1, 30:1, 20:1, 10:1, 8:2, 7:3, the chloroform of 6:4 and 0:1 and the mixed solution of acetone are as moving phase, the elution volume of each gradient is 1500 milliliters), flow velocity during wash-out is 10ml per minute.Collect and adopt volume ratio to be the front 500 milliliter elutriants (be designated as solution first) of chloroform-acetone as moving phase of 50:1, collect adopt volume ratio be 20:1 chloroform-acetone as moving phase rear 500 milliliters of elutriants and adopt volume ratio to be the mixed solution (be designated as solution second) of chloroform-acetone as front 500 milliliters of elutriants of moving phase of 10:1.
By solution first in 38 DEG C of concentrating under reduced pressure, vacuum tightness <3mmHg vacuum-drying, obtains dry-matter first (2.0g).By 2.0g dry-matter first, use 3ml dissolve with methanol, the centrifugal 5min of 5000rpm, collect supernatant liquor, supernatant liquor is splined on gel column sephadexLH-20(Φ 2.5 × 120cm, sephadexLH-20, Merck company), then adopt methyl alcohol to carry out wash-out as moving phase, flow velocity during wash-out is 1ml per minute.Collect the elutriant of the 140th milliliter-180 milliliters, in 38 DEG C of concentrating under reduced pressure, vacuum tightness <3mmHg vacuum-drying, obtains compound shown in formula I-1; Collect the elutriant of the 200th milliliter-240 milliliters, in 38 DEG C of concentrating under reduced pressure, vacuum tightness <3mmHg vacuum-drying, obtains compound shown in formula I-2; Collect the elutriant of the 260th milliliter-340 milliliters, in 38 DEG C of concentrating under reduced pressure, vacuum tightness <3mmHg vacuum-drying, obtains compound shown in formula I-3.
By solution second in 38 DEG C of concentrating under reduced pressure, vacuum tightness <3mmHg vacuum-drying, obtains dry-matter fourth (390.5mg).By 390.5mg dry-matter fourth, use 1.5ml dissolve with methanol, the centrifugal 5min of 5000rpm, collect supernatant liquor, by supernatant liquor and the reverse silica gel (YMC*GEL of 400mgODS, 12nm, S-50 μm) mixing, evaporate to dryness organic solvent in 46 DEG C of water-baths, then mesolow chromatographic column (Φ 3.0 × 30cm is added into, be filled with 40gODS reverse phase silica gel, YMC*GEL, 12nm, S-50 μm) in, then methanol-water is adopted to carry out gradient elution (in gradient elution process as moving phase, volume ratio is adopted to be 20:80 successively, 30:70, 40:60, 50:50, 60:40, the mixed solution of the first alcohol and water of 85:15 and 0:1 is as moving phase, the elution volume of each gradient is 200 milliliters), flow velocity during wash-out is 3ml per minute.The volume ratio of collecting first alcohol and water is the elutriant of latter 100 milliliters of the moving phase of 40:60, and in 38 DEG C of concentrating under reduced pressure, vacuum tightness <3mmHg vacuum-drying, obtains compound shown in formula I-4; The volume ratio of collecting first alcohol and water is the elutriant of latter 100 milliliters of the moving phase of 50:50, and in 38 DEG C of concentrating under reduced pressure, vacuum tightness <3mmHg vacuum-drying, obtains compound shown in formula II-1; The volume ratio of collecting first alcohol and water is the elutriant of latter 100 milliliters of the moving phase of 60:40, and in 38 DEG C of concentrating under reduced pressure, vacuum tightness <3mmHg vacuum-drying, obtains compound shown in formula II-2; The volume ratio of collecting first alcohol and water is the elutriant of latter 100 milliliters of the moving phase of 85:15, and in 38 DEG C of concentrating under reduced pressure, vacuum tightness <3mmHg vacuum-drying, obtains compound shown in formula II-3.
Five, the structural confirmation of compound
Compound shown in formula I-1 (called after sterenolA), its physical and chemical parameter and spectral data as follows:
Brown unformed powder; Uv-absorbing (methyl alcohol) λ
maxnm (log ε): 212 (6.51) nm, 266 (4.21) nm; Infrared absorption ν
max(methyl alcohol) 3360,3071,2979,2938,1700,1685,1641,1617,1585,1441,1310,1284,1157,1096,846cm
-1; HRTOFMS (positive ion mode) m/z:[M+Na]
+409.1258 [calcd.forC
21h
22o
7na (M+Na)
+, 409.1258]; Its molecular formula is C
21h
22o
7.
1h-NMR (500MHz, methyl alcohol-d
4): δ 6.53 (1H, s, H-2 '), δ 6.30 (1H, s, H-5), δ 6.22 (1H, d, J=2.3Hz, H-3), δ 5.04 (1H, t, J=6.6Hz, H-2 ' '), δ 3.27 (2H, d, J=6.6Hz, H-1 ' '), δ 2.57 (6H, s, H-7 ', H-8), δ 1.56 (3H, s, H-5 ' '), δ 1.50 (3H, s, H-4 ' ').
13c-NMR (125MHz, methyl alcohol-d
4): δ 175.2 (C-8 '), δ 171.1 (C-7), δ 166.9 (C-2), δ 164.7 (C-4), δ 164.0 (C-5 '), δ 153.6 (C-1 '), δ 144.9 (C-6), δ 141.8 (C-3 '), δ 132.6 (C-3 ' '), δ 122.8 (C-2 ' '), δ 121.0 (C-6 '), δ 117.7 (C-2 '), δ 112.9 (C-5), δ 111.6 (C-4 '), δ 105.1 (C-1), δ 101.8 (C-3), δ 25.7 (C-5 ' '), δ 24.5 (C-8), δ 23.9 (C-7 '), δ 23.8 (C-1 ' '), δ 17.7 (C-4 ' ').
Compound shown in formula I-2 (called after sterenolB), its physical and chemical parameter and spectral data as follows:
Brown unformed powder; Uv-absorbing (methyl alcohol) λ
maxnm (log ε): 213 (6.23) nm, 265 (4.51) nm; Infrared absorption ν
max(methyl alcohol) 3370,2928,2855,1663,1617,1443,1403,1377,1310,1250,1195,1149,1095,806cm
-1; HRTOFMS (positive ion mode) m/z:[M+Na]
+423.1419 [calcd.forC
22h
24o
7na (M+Na)
+, 423.1414]; Its molecular formula is C
22h
24o
7.
1h-NMR (500MHz, chloroform-d
1): δ 6.50 (1H, s, H-2 '), δ 6.32 (1H, s, H-3), δ 6.31 (1H, s, H-5), δ 5.07 (1H, t, J=6.7Hz, H-2 ' '), δ 3.97 (3H, s, H-9 '), δ 3.30 (2H, d, J=6.7Hz, H-1 ' '), δ 2.62 (3H, s, H-8), δ 2.53 (3H, s, H-7 '), δ 1.59 (3H, s, H-5 ' '), δ 1.55 (3H, s, H-4 ' ').
13c-NMR (125MHz, methyl alcohol-d
4): δ 172.4 (C-8 '), δ 170.2 (C-7), δ 166.5 (C-2), δ 161.2 (C-4), δ 162.8 (C-5 '), δ 152.4 (C-1 '), δ 144.7 (C-6), δ 140.2 (C-3 '), δ 132.6 (C-3 ' '), δ 121.3 (C-2 ' '), δ 120.5 (C-6 '), δ 116.9 (C-2 '), δ 112.0 (C-5), δ 110.5 (C-4 '), δ 104.9 (C-1), δ 101.7 (C-3), δ 52.4 (C-9 '), δ 25.7 (C-5 ' '), δ 24.7 (C-8), δ 24.2 (C-7 '), δ 23.3 (C-1 ' '), δ 17.8 (C-4 ' ').
Compound shown in formula I-3 (called after sterenolC), its physical and chemical parameter and spectral data as follows:
Brown unformed powder; Uv-absorbing (methyl alcohol) λ
maxnm (log ε): 217 (6.23) nm, 260 (4.51) nm; Infrared absorption ν
max(methyl alcohol) 3375,2979,2881,1719,1653,1640,1617,1444,1406,1377,1313,1240,1197,1163,1095,846cm
-1; HRTOFMS (positive ion mode) m/z:[M+Na]
+371.1489 [calcd.forC
21h
22o
6h (M+H)
+, 371.1489]; Its molecular formula is C
21h
22o
6.
1h-NMR (500MHz, chloroform-d
1): δ 6.51 (1H, s, H-2 '), δ 6.32 (1H, s, H-5), δ 6.33 (1H, d, H-3), δ 5.05 (1H, t, J=6.6Hz, H-2 ' '), δ 3.29 (2H, d, J=6.6Hz, H-1 ' '), δ 2.62 (3H, s, H-8), δ 2.60 (3H, s, H-7 '), δ 1.58 (3H, s, H-5 ' '), δ 1.55 (3H, s, H-4 ' ').
13c-NMR (125MHz, methyl alcohol-d
4): δ 194.7 (C-8 '), δ 170.0 (C-7), δ 166.5 (C-2), δ 161.3 (C-4), δ 163.5 (C-5 '), δ 154.8 (C-1 '), δ 144.6 (C-6), δ 141.0 (C-3 '), δ 133.0 (C-3 ' '), δ 122.6 (C-2 ' '), δ 121.5 (C-6 '), δ 116.4 (C-2 '), δ 112.1 (C-5), δ 116.8 (C-4 '), δ 104.7 (C-1), δ 101.7 (C-3), δ 25.7 (C-5 ' '), δ 24.7 (C-8), δ 18.3 (C-7 '), δ 22.6 (C-1 ' '), δ 17.8 (C-4 ' ').
Compound shown in formula I-4 (called after sterenolD), its physical and chemical parameter and spectral data as follows:
Colourless oil liquid; Uv-absorbing (methyl alcohol) λ
maxnm (log ε): 205 (6.44) nm, 270 (4.23) nm; Infrared absorption ν
max(methyl alcohol) 3306,2971,2931,1658,1621,1586,1445,1426,1376,1312,1254,1198,1166,1097,1009,845,792,697cm
-1; HRTOFMS (positive ion mode) m/z:[M+Na]
+425.1569 [calcd.forC
22h
26o
7na (M+Na)
+, 425.1571]; Its molecular formula is C
22h
26o
7.
1h-NMR (500MHz, chloroform-d
1): δ 6.67 (1H, s, H-2 '), δ 6.31 (1H, s, H-3), δ 6.30 (1H, s, H-5), δ 5.11 (1H, t, J=6.8Hz, H-2 ' '), δ 3.49 (3H, s, H-9 '), δ 3.30 (2H, d, J=6.8Hz, H-1 ' '), δ 2.62 (3H, s, H-8), δ 4.62 (2H, s, H-7 '), δ 4.81 (2H, s, H-8 '), δ 1.62 (3H, s, H-5 ' '), δ 1.59 (3H, s, H-4 ' ').
13c-NMR (125MHz, methyl alcohol-d
4): δ 170.6 (C-7), δ 166.4 (C-2), δ 161.1 (C-4), δ 156.4 (C-5 '), δ 148.5 (C-1 '), δ 144.6 (C-6), δ 137.6 (C-3 '), δ 133.5 (C-3 ' '), δ 121.3 (C-2 ' '), δ 121.7 (C-6 '), δ 114.2 (C-2 '), δ 111.9 (C-5), δ 119.1 (C-4 '), δ 104.9 (C-1), δ 101.6 (C-3), δ 58.6 (C-9 '), δ 69.3 (C-8 '), δ 25.7 (C-5 ' '), δ 24.7 (C-8), δ 63.6 (C-7 '), δ 23.6 (C-1 ' '), δ 17.8 (C-4 ' ').
Compound shown in formula II-1 (called after StereninE), its physical and chemical parameter and spectral data as follows:
White powder; Uv-absorbing (methyl alcohol) λ
maxnm (log ε): 218 (6.08) nm, 275 (3.41) nm; Infrared absorption ν
max(methyl alcohol) 3202,2972,2930,1709,1661,1601,1447,1311,1254,1200,1162,1104,1058,846,792,723cm
-1; HRTOFMS (positive ion mode) m/z:[M+H]
+470.1809 [calcd.forC
25h
27nO
8h (M+H)
+, 470.1809]; Its molecular formula is C
25h
27nO
8.
1h-NMR (500MHz, methyl alcohol-d
4): δ 7.06 (1H, 2, H-7), δ 6.31 (1H, s, H-5 ' '), δ 6.23 (1H, d, J=2.3Hz, H-3 ' '), δ 5.03 (1H, t, J=6.6Hz, H-2 '), δ 4.46 (2H, d, J=9.3Hz, H-3), δ 3.69 (2H, t, J=6.9Hz, H-1 ' ' '), δ 3.38 (2H, d, J=6.6Hz, H-1 '), δ 2.60 (3H, s, H-8 ' '), δ 2.38 (2H, t, J=7.0Hz, H-3 ' ' '), δ 2.02 (2H, m, H-2 ' ' '), δ 1.56 (3H, s, H-5 '), δ 1.50 (3H, s, H-4 ').
13c-NMR (125MHz, methyl alcohol-d
4): δ 176.8 (C-4 ' ' '), δ 171.8 (C-7 ' '), δ 170.4 (C-1), δ 167.0 (C-2 ' '), δ 164.7 (C-4 ' '), δ 152.1 (C-4), δ 151.2 (C-6), δ 144.9 (C-6 ' '), δ 133.1 (C-3 '), δ 132.5 (C-7a), δ 127.1 (C-5), δ 126.7 (C-3a), δ 122.5 (C-2 '), δ 112.9 (C-5 ' '), δ 110.0 (C-7), δ 105.1 (C-1 ' '), δ 101.8 (C-3 ' '), δ 49.5 (C-3), δ 43.2 (C-1 ' ' '), δ 32.2 (C-3 ' ' '), δ 25.7 (C-5 '), δ 24.7 (C-2 ' ' '), δ 24.6 (C-1 '), δ 24.5 (C-8 ' '), δ 17.7 (C-4 ').
Compound shown in formula II-2 (called after StereninF), its physical and chemical parameter and spectral data as follows:
White powder; Optical value [α] 20D-12.05 (c0.2, methyl alcohol); Uv-absorbing (methyl alcohol) λ
maxnm (log ε): 213 (4.28) nm, 263 (2.52) nm; Infrared absorption ν
max(methyl alcohol) 3193,2964,2925,2874,1718,1658,1623,1601,1455,1310,1253,1199,1160,1093,1063,846,791cm
-1; HRTOFMS (positive ion mode) m/z:[M+Na]
+520.1940 [calcd.forC
27h
31nO
8na (M+Na)
+, 520.1942]; Its molecular formula is C
27h
31nO
8.
1h-NMR (500MHz, methyl alcohol-d
4): δ 7.08 (1H, 2, H-7), δ 6.31 (1H, s, H-5 ' '), δ 6.23 (1H, d, J=2.3Hz, H-3 ' '), δ 5.04 (1H, t, J=6.6Hz, H-2 '), δ 4.45 (1H, d, J=17.6Hz, H-3), δ 4.77 (1H, d, J=17.6Hz, H-3), δ 4.74 (1H, d, J=10.4Hz, H-1 ' ' '), δ 3.39 (2H, d, J=6.6Hz, H-1 '), δ 2.59 (3H, s, H-8 ' '), δ 1.19 (1H, m, H-3 ' ' '), δ 1.44 (1H, m, H-3 ' ' '), δ 2.20 (1H, m, H-2 ' ' '), δ 1.57 (3H, s, H-5 '), δ 1.51 (3H, s, H-4 '), δ 0.95 (3H, t, J=7.3Hz, H-4 ' ' '), δ 1.09 (3H, d, J=6.6Hz, H-5 ' ' ').
13c-NMR (125MHz, methyl alcohol-d
4): δ 174.5 (C-6 ' ' '), δ 171.6 (C-7 ' '), δ 170.4 (C-1), δ 166.9 (C-2 ' '), δ 164.7 (C-4 ' '), δ 152.2 (C-4), δ 151.2 (C-6), δ 144.9 (C-6 ' '), δ 133.2 (C-3 '), δ 131.7 (C-7a), δ 127.3 (C-5), δ 127.0 (C-3a), δ 122.5 (C-2 '), δ 112.9 (C-5 ' '), δ 110.2 (C-7), δ 105.1 (C-1 ' '), δ 101.8 (C-3 ' '), δ 47.1 (C-3), δ 60.8 (C-1 ' ' '), δ 26.6 (C-3 ' ' '), δ 25.7 (C-5 '), δ 36.2 (C-2 ' ' '), δ 24.5 (C-1 '), δ 24.6 (C-8 ' '), δ 17.7 (C-4 '), δ 16.2 (C-5 ' ' '), δ 11.1 (C-4 ' ' ').
Compound shown in formula II-3 (called after StereninG), its physical and chemical parameter and spectral data as follows:
White powder; Optical value [α] 20D-11.33 (c0.3, methyl alcohol); Uv-absorbing (methyl alcohol) λ
maxnm (log ε): 213 (6.18) nm, 264 (2.45) nm; Infrared absorption ν
max(methyl alcohol) 3368,2923,2869,1711,1654,1637,1599,1450,1310,1253,1198,1158,1093,1071,844,795,725cm
-1; HRTOFMS (positive ion mode) m/z:[M+H]
+498.2124 [calcd.forC
27h
31nO
8h (M+H)
+, 498.2122]; Its molecular formula is C
27h
31nO
8.
1h-NMR (500MHz, methyl alcohol-d
4): δ 7.09 (1H, 2, H-7), δ 6.31 (1H, s, H-5 ' '), δ 6.23 (1H, d, J=2.3Hz, H-3 ' '), δ 5.04 (1H, t, J=6.6Hz, H-2 '), δ 4.40 (1H, d, J=17.6Hz, H-3), δ 4.63 (1H, d, J=17.6Hz, H-3), δ 5.06 (1H, m, H-1 ' ' '), δ 3.39 (2H, d, J=6.6Hz, H-1 '), δ 2.61 (3H, s, H-8 ' '), δ 1.52 (1H, m, H-3 ' ' '), δ 1.99 (1H, m, H-2 ' ' '), δ 1.90 (1H, m, H-2 ' ' '), δ 1.57 (3H, s, H-5 '), δ 1.51 (3H, s, H-4 '), δ 1.00 (3H, d, J=4.9Hz, H-4 ' ' '), δ 1.01 (3H, d, J=5.0Hz, H-5 ' ' ').
13c-NMR (125MHz, methyl alcohol-d
4): δ 174.5 (C-6 ' ' '), δ 171.5 (C-7 ' '), δ 170.4 (C-1), δ 166.9 (C-2 ' '), δ 164.7 (C-4 ' '), δ 152.2 (C-4), δ 151.2 (C-6), δ 144.9 (C-6 ' '), δ 133.2 (C-3 '), δ 131.9 (C-7a), δ 127.3 (C-5), δ 127.0 (C-3a), δ 122.5 (C-2 '), δ 112.9 (C-5 ' '), δ 110.2 (C-7), δ 105.1 (C-1 ' '), δ 101.8 (C-3 ' '), δ 46.4 (C-3), δ 53.6 (C-1 ' ' '), δ 26.3 (C-3 ' ' '), δ 25.7 (C-5 '), δ 39.4 (C-2 ' ' '), δ 24.5 (C-1 '), δ 24.6 (C-8 ' '), δ 17.7 (C-4 '), δ 23.5 (C-5 ' ' '), δ 21.3 (C-4 ' ' ').
The vitro inhibition alpha-glucosidase activity test of embodiment 2, embodiment 1 gained compound
Test sample solution: 7 kinds of compound solutions that the embodiment 1 that final concentration is respectively 100.0 μMs, 50.0 μMs, 25.0 μMs, 12.5 μMs, 6.25 μMs, 3.13 μMs prepares are (after being dissolved in a small amount of DMSO, with distilled water diluting to respective concentration, the final volume mark <0.1% of control DMSO); And be respectively the acarbose solution of 4000 μMs, 1000 μMs, 250 μMs, 62.5 μMs (after being dissolved in a small amount of DMSO as the final concentration of positive control, with distilled water diluting to respective concentration, the final volume mark <0.1% of control DMSO).
Get the above test sample solution of 25 μ L different concns, add 25 μ L alpha-glucosaccharase enzyme aqueous solution (concentration is 0.2U/mL) and 175 μ L phosphate buffer soln (50mM, pH7.0).Mixing solutions adds the 25 μ L4-oil of mirbane-α-D-glucopyranoside aqueous solution (concentration is 2.5mM) after room temperature places 10min, 37 DEG C of constant-temperature incubation 30min, the centrifugal 5min of 5000rpm, gets in supernatant 150 μ L to 96 orifice plate, measures light absorption value at wavelength 405nm place.
Experiment arranges blank group simultaneously: use the phosphate buffered saline buffer (50mM, pH7.0) of 25 μ L to replace test sample solution; Blank group: use the phosphate buffered saline buffer (50mM, pH7.0) of 25 μ L to replace test sample solution, the phosphate buffered saline buffer (50mM, pH7.0) of 25 μ L replaces alpha-glucosaccharase enzyme aqueous solution simultaneously; Sample controls group: use the phosphate buffered saline buffer (50mM, pH7.0) of 25 μ L to replace alpha-glucosaccharase enzyme aqueous solution.
Above-mentioned each experimental group all in triplicate, results averaged.Use following formulae discovery sample to the inhibiting rate of alpha-glucosidase:
Inhibiting rate (%)=[1-(sample sets light absorption value-sample controls group light absorption value)/(blank group light absorption value-blank group light absorption value)] × 100%
To experimental data statistical study, use IC
50the IC of each test sample of computed in software
50value result is as shown in table 1.Visible, the intensity of the Inhibiting α-glucosidase of compound shown in formula I-1 is 84 times of positive control medicine acarbose, the intensity of the Inhibiting α-glucosidase of compound shown in formula I-2 is 209 times of positive control medicine acarbose, the intensity of the Inhibiting α-glucosidase of compound shown in formula I-3 is 106 times of positive control medicine acarbose, the intensity of the Inhibiting α-glucosidase of compound shown in formula I-4 is 28 times of positive control medicine acarbose, the intensity of the Inhibiting α-glucosidase of compound shown in formula II-1 is 17 times of positive control medicine acarbose, the intensity of the Inhibiting α-glucosidase of compound shown in formula II-2 is 49 times of positive control medicine acarbose, the intensity of the Inhibiting α-glucosidase of compound shown in formula II-3 is 23 times of positive control medicine acarbose.
The alpha-glucosaccharase enzyme inhibition activity detected result of table 17 kind of compound
The experimentation on animals of embodiment 3, embodiment 1 gained compound
One, the structure of Rat Type II diabetes rat model
1, laboratory animal
Get wistar male rat 50, adaptability raises one week: room temperature 18-25 DEG C, humidity 50-60%, in 12/12 hour light and shade cycle, freely ingests, drinks water; Give standard rat chow.
2, type ii diabetes rat model modeling experiment
Blank group: 10 wistar male rats, give common standard rat chow.
Model group: 90 wistar male rats, give high glucose and high fat feed.
After blank group and model group rats are all fed 4 weeks, fasting 8 hours, got rat tail vein blood and measures fasting plasma glucose in the 29th day, and intravenous injection U-9889 45mg/kg body weight, feed after intravenous injection 72 hours, fasting 8 hours, gets rat tail vein blood and surveys fasting plasma glucose.
Result shows, and after injection U-9889, the fasting blood sugar of model group rats is 23.17 ± 0.51mmol/L, is significantly higher than 5.34 ± 0.61mmol/L(P < 0.01 of blank group rat).Model group rats occurs that the diabetic symptoms such as many drinks, diuresis, many foods are bright simultaneously.These results suggest that type ii diabetes rat model modeling success.
Two, administration experiment
Administration experiment grouping is as shown in table 2, and wherein, model group (test group) is divided into 9 groups at random, often organizes the type ii diabetes rat that 10 steps one successfully construct; Blank group is 10 normal wistar male rats.
Table 2 administration experiment grouping and process
Note: 7 kinds of compounds and acarbose are all dissolved in distilled water.
Each group of rat is with feeding corresponding feed under condition, and gastric infusion, once a day, successive administration one week, each group of rat is got in fasting tail vein after 8 hours measures fasting plasma glucose.
Test in triplicate, results averaged.
Result is as shown in table 3, visible, and 7 kinds of compounds have the effect significantly controlling type II diabetes rat model blood glucose value, and comparatively positive drug acarbose is stronger for its action effect.
Table 3 is group rat fasting blood-glucose value respectively
| Experimental group | Fasting plasma glucose mmol/L | P value |
| Formula I-1Sterenol A | 6.97±0.78 | <0.01 |
| Formula I-2Sterenol B | 4.23±0.38 | <0.01 |
| Formula I-3Sterenol C | 6.22±0.45 | <0.01 |
| Formula I-4Sterenol D | 7.89±0.28 | <0.01 |
| Formula II-1Sterenin E | 8.06±0.84 | <0.01 |
| Formula II-2Sterenin F | 7.56±0.76 | <0.01 |
| Formula II-3Sterenin G | 7.77±0.14 | <0.01 |
| Positive drug group | 12.49±0.59 | <0.01 |
| Model control group | 22.06±0.68 | - |
| Blank group | 4.82±0.93 | <0.01 |
Claims (11)
1. compound, shown in I or formula II:
In described formula I, R
1and R
2for arbitrary in following (a)-(d):
(a) R
1=CH
3, R
2=COOH; Corresponding compound is designated as compound S terenolA;
(b) R
1=CH
3, R
2=COOCH
3; Corresponding compound is designated as compound S terenolB;
(c) R
1=CH
3, R
2=CHO; Corresponding compound is designated as compound S terenolC;
(d) R
1=CH
2oH, R
2=CH
2oCH
3; Corresponding compound is designated as compound S terenolD;
In described formula II, R
1for arbitrary in following (e)-(g):
(e) R
1=CH
2cH
2cH
2cOOH; Corresponding compound is designated as compound S tereninE;
(f) R
1=CH (COOH) CH (CH
3) CH
2cH
3; Corresponding compound is designated as compound S tereninF;
(g) R
1=CH (COOH) CH
2cH (CH
3) CH
3; Corresponding compound is designated as compound S tereninG.
2. the application of compound or pharmaceutically acceptable salt thereof described in claim 1 in following (A1)-(A4) is arbitrary:
(A1) alpha-glucosidase inhibitor is prepared;
(A2) preparation treats and/or prevents the medicine of type ii diabetes;
(A3) medicine of Inhibiting α-glucosidase is prepared;
(A4) preparation has food or the functional health care product of blood sugar reducing function.
3. treat and/or prevent the medicine of type ii diabetes, its activeconstituents is compound or pharmaceutically acceptable salt thereof described in claim 1.
4. the medicine of Inhibiting α-glucosidase, its activeconstituents is compound or pharmaceutically acceptable salt thereof described in claim 1.
5. have food or the functional health care product of blood sugar reducing function, its activeconstituents is compound or pharmaceutically acceptable salt thereof described in claim 1.
6. the preparation method of compound described in claim 1, comprises the steps:
(1) thick hair Boreostereum vibrans (Stereumhirsutum) is inoculated on solid medium carries out solid fermentation cultivation;
(2) in the system after step (1) fermentation, add the extracting solution that organic solvent soak extraction obtains containing compound shown in compound and formula II shown in claim 1 Chinese style I, by described extracting solution in 36-40 DEG C of concentrating under reduced pressure, vacuum-drying, obtains the crude extract containing compound shown in compound and described formula II shown in described formula I.
7. method according to claim 6, is characterized in that: in step (1), and described solid medium is following a1) or a2):
A1) substratum be made up of 1 weight part base material and 1-1.5 weight parts water;
A2) be made up of 1 weight part base material, 1-1.5 weight parts water and 0.02-0.1 weight part sugar;
Described base material is at least one in long-grained nonglutinous rice, wheat, Gorgon fruit, oat, polished rice, corn, glutinous rice and the seed of Job's tears; Described sugar is glucose;
In step (1), the temperature that described solid fermentation is cultivated is 22-28 DEG C, and mode is lucifuge quiescent culture, and the time is 10-50 days.
8. the method according to claim 6 or 7, is characterized in that: in step (2), and described organic solvent is ethyl acetate.
9. the method according to claim 6 or 7, it is characterized in that: in step (2), the method for the extracting solution that described soak extraction obtains containing compound shown in compound and formula II shown in claim 1 Chinese style I comprises following b1)-b4) step:
B1) first time soak extraction: add ethyl acetate and soak 7 days in the system after step (1) fermentation, period every 8h supersound process once, gets supernatant liquor after immersion;
B2) second time soak extraction: to step b1) residuum in add ethyl acetate and soak 7 days, period every 8h supersound process once, gets supernatant liquor after immersion;
B3) third time soak extraction: to step b2) residuum in add ethyl acetate and soak 7 days, period every 8h supersound process once, gets supernatant liquor after immersion;
B4) combining step b1)-b3) supernatant liquor, namely obtain the described extracting solution containing compound shown in compound and described formula II shown in formula I.
10. the method according to claim 6 or 7, is characterized in that: described method also comprises the steps (3)-(5):
(3) the described crude extract containing compound shown in compound with formula II shown in formula I step (2) obtained carries out silica gel column chromatography and is separated, using the mixed solution of chloroform and acetone as moving phase, the gradient being followed successively by 100:0,100:1,50:1,30:1,20:1,10:1,8:2,7:3,6:4 and 0:1 according to the volume ratio of chloroform in moving phase and acetone carries out gradient elution, and the elution volume of the moving phase of each gradient is 1500 milliliters; The volume ratio of collecting chloroform and acetone is the elutriant of first 500 milliliters of the moving phase of 50:1, is designated as solution first; The volume ratio of collecting chloroform and acetone is the elutriant of latter 500 milliliters of the moving phase of 20:1, and the volume ratio of chloroform and acetone is the elutriant of first 500 milliliters of the moving phase of 10:1, and after mixing, solution is designated as solution second;
(4) the solution first that step (3) obtains is carried out gel permeation chromatography, carry out wash-out using methyl alcohol as moving phase; Collect the elutriant of the 140th milliliter-180 milliliters, concentrated in 36-40 DEG C of pressure, vacuum-drying, obtains the terenolA of compound S described in claim 1; Collect the elutriant of the 200th milliliter-240 milliliters, in 36-40 DEG C of concentrating under reduced pressure, vacuum-drying, obtains the terenolB of compound S described in claim 1; Collect the elutriant of the 260th milliliter-340 milliliters, in 36-40 DEG C of concentrating under reduced pressure, vacuum-drying, obtains the terenolC of compound S described in claim 1;
(5) the solution second that step (3) obtains is carried out ODS reversed-phase silica gel chromatography, using the mixed solution of first alcohol and water as moving phase, the gradient being followed successively by 20:80,30:70,40:60,50:50,60:40,85:15 and 0:1 according to the volume ratio of first alcohol and water in moving phase carries out gradient elution, and the elution volume of the moving phase of each gradient is 200 milliliters; The volume ratio of collecting first alcohol and water is the elutriant of latter 100 milliliters of the moving phase of 40:60, and in 36-40 DEG C of concentrating under reduced pressure, vacuum-drying, obtains the terenolD of compound S described in claim 1; The volume ratio of collecting first alcohol and water is the elutriant of latter 100 milliliters of the moving phase of 50:50, and in 36-40 DEG C of concentrating under reduced pressure, vacuum-drying, obtains the tereninE of compound S described in claim 1; The volume ratio of collecting first alcohol and water is the elutriant of latter 100 milliliters of the moving phase of 60:40, and in 36-40 DEG C of concentrating under reduced pressure, vacuum-drying, obtains the tereninF of compound S described in claim 1; The volume ratio of collecting first alcohol and water is the elutriant of latter 100 milliliters of the moving phase of 85:15, and in 36-40 DEG C of concentrating under reduced pressure, vacuum-drying, obtains the tereninG of compound S described in claim 1.
11. methods according to claim 6 or 7, is characterized in that: described thick hair Boreostereum vibrans (Stereumhirsutum) is thick hair Boreostereum vibrans (Stereumhirsutum) THG20.
Priority Applications (1)
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| Title |
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| "Studies on Glyoxalase inhibitor isolation of a new active agent, MS-3, from a mushroom culture";Kurasawa, Shyogo等;《Agricultural and Biological Chemistry》;19751231;第39卷(第10期);第2003-2008页 * |
| "The crystal and molecular structure of a dibromo-derivative of MS-3: a glyoxalase I inhibitor produced by a mushroom, Stereum hirsutum";Nakamura, Hikaru等;《Agricultural and Biological Chemistry》;19761231;第40卷(第9期);第1781-1784页 * |
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