CN104987319A - Marine fungus-derived depside compounds and application thereof in treatment of type 2 diabetes - Google Patents
Marine fungus-derived depside compounds and application thereof in treatment of type 2 diabetes Download PDFInfo
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- CN104987319A CN104987319A CN201510205871.1A CN201510205871A CN104987319A CN 104987319 A CN104987319 A CN 104987319A CN 201510205871 A CN201510205871 A CN 201510205871A CN 104987319 A CN104987319 A CN 104987319A
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- depsidcs
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D321/00—Heterocyclic compounds containing rings having two oxygen atoms as the only ring hetero atoms, not provided for by groups C07D317/00 - C07D319/00
- C07D321/02—Seven-membered rings
- C07D321/10—Seven-membered rings condensed with carbocyclic rings or ring systems
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P17/00—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms
- C12P17/02—Oxygen as only ring hetero atoms
- C12P17/08—Oxygen as only ring hetero atoms containing a hetero ring of at least seven ring members, e.g. zearalenone, macrolide aglycons
Abstract
The invention belongs to the field of medicinal compounds, and concretely discloses marine fungus-derived depside compounds and an application thereof in treatment of type 2 diabetes. The structure of the depside compounds is represented by formula (I). The compounds 1-6 can substantially inhibit the activity of alpha-glucosidase, the IC50 value of the compound 1 is 10.4[mu]M, the IC50 value of the compound 2 is 13.3[mu]M, the IC50 value of the compound 3 is 2.1[mu]M, the IC50 value of the compound 4 is 12.4[mu]M, the IC50 value of the compound 5 is 9.8[mu]M, and the IC50 value of the compound 6 is 11.7[mu]M, wherein the IC50 value of positive control acarbose is 553.7[mu]M. Kinetic researches of the alpha-glucosidaseinhibition effect show that the compounds 1, 3 and 5 are noncompetitive inhibitors. The compounds can be used for preparing alpha-glucosidase inhibitor medicines in order to prevent and treat the type 2 diabetes.
Description
Technical field
The present invention relates to medical compounds field, be specifically related to the medical compounds in thalassiomycetes source, more specifically, relate to the depsidcs in a class thalassiomycetes source and the application for the treatment of type ii diabetes thereof.
Background technology
At present, the type ii diabetes number of patients in global range is more than 3.7 hundred million people, and the number of patient also increases fast.This growth changes with Economic development, aging population, day by day urbanization, food habits, physical exertion reduces relevant with other living-pattern preservation.Thus, type ii diabetes is considered to one of most stern challenge of 21 century world community public health security.In type ii diabetes, body can produce Regular Insulin, but because of insulin secretion relative deficiency or effect defect (also referred to as insulin resistant), causes blood sugar increasing.And alpha-glucosidase is a kind of important target of the early stage type ii diabetes for the treatment of.It is the general name of the enzyme that a class can be alpha-glucose-based from the non-reducing end catalytic hydrolysis containing α-glucose glycosidic bond substrate.Alpha-glucosidase is distributed widely in organism, participates in the biosynthesizing of food digestion, glycoprotein, many bioprocesss such as the composition and decomposition metabolism of polysaccharide and saccharide complex.Alpha-glucosidase inhibitor is a class reaches treatment diabetes oral antidiabetic drug to delay enteron aisle carbohydrate absorption, its action principle is: competitive inhibition is positioned at the various alpha-glucosidases of small intestine, the speed being decomposed into glucose is slowed down, thus slow down the absorption of glucose in enteron aisle, improve the peak of postprandial blood sugar.Research confirms, alpha-glucosidase inhibitor can be prevented and treated post prandial hyperglycemia and alleviate hyperinsulinemia, can improve sugar tolerance simultaneously.Current, the alpha-glucosidase inhibitor of clinical application mainly contains acarbose and voglibose.In order to avoid or reduce the adverse side effect of current medical or resistance, provide more drug candidates, it is very necessary for therefore finding new alpha-glucosidase inhibitor and developing into medicine simultaneously.
Summary of the invention
The object of the invention is to the defect existed for current type ii diabetes medicine, the depsidcs in a class thalassiomycetes source is provided.
Another object of the present invention is to provide the preparation method of the depsidcs in a class thalassiomycetes source.
The application of depsidcs in preparation treatment type ii diabetes medicine being to provide a class thalassiomycetes source an object of the present invention.
To achieve these goals, the present invention is achieved by the following technical programs:
The depsidcs in one class thalassiomycetes source, its structural formula is as shown in formula I:
Described depsidcs 1 – 6 is separated to obtain from the fermented liquid of thalassiomycetes Meyerozyma sp.HZ-Y2; Described thalassiomycetes Meyerozyma sp.HZ-Y2 is deposited in China typical culture collection center on April 6th, 2015, and preservation address is Wuhan University of Wuhan, China city, and deposit number is CCTCCNO:M 2015203.
Preferably, the preparation method of above-mentioned depsidcs is as follows:
S1. by thalassiomycetes Meyerozyma sp.HZ-Y
2access seed culture medium, shaking table is cultivated, and obtains seed culture fluid;
S2. seed culture fluid is accessed in fermention medium, quiescent culture;
S3. filtered by tunning and obtain thalline, thalline methyl alcohol soaks, and concentrating under reduced pressure obtains medicinal extract, then through chromatographic separation, obtains depsidcs 1 – 6.
As a kind of preferred version, in above-mentioned preparation method, the component of seed culture medium described in step S1 is: potato 200g, glucose 20g, water 1L.
As a kind of preferred version, in above-mentioned preparation method, the component of fermention medium described in step S2 is: northeast rice 7000g, sea salt 210g, water 7L.
As a kind of preferred version, in above-mentioned preparation method, described in step S1, shaking table culture condition is: rotating speed 200rpm, temperature 28 DEG C, incubation time 72h.
As a kind of preferred version, in above-mentioned preparation method, quiescent culture temperature described in step S2 is 25 DEG C, and incubation time is 28 days.
As a kind of preferred version, in above-mentioned preparation method, described in step S3, medicinal extract silica gel column chromatography is separated, and is separated the ethyl acetate/petroleum ether gradient elution with 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 100%.
Medicinal extract is when chromatographic separation, and the 6th component adopts dextrane gel Sephadex LH-20 chromatography, is that eluent carries out wash-out with the methanol-chloroform that volume ratio is 1:1, recycle silicon glue post layer, and 25% ethyl acetate-light petrol is eluent, obtains compound 2 and 6.8th component employing dextrane gel Sephadex LH-20 chromatography is methyl alcohol by volume ratio is that eluent carries out wash-out, and recycle silicon glue post layer, 30% ethyl acetate-light petrol is eluent, obtains compound 3 and 4.15th component adopts dextrane gel Sephadex LH-20 chromatography, is that eluent carries out wash-out, then prepares compound 1 and 5 with partly preparing RPLC with the methanol-chloroform that volume ratio is 1:1.
Depsidcs 1-6 of the present invention has restraining effect to alpha-glucosidase, can be used for preparing alpha-glucosidase inhibitor medicament, treatment type ii diabetes.Therefore, the application of application claims protection depsidcs 1-6 in preparation treatment type ii diabetes medicine.In addition, the present invention also protects depsidcs 1-6 preparing the application in alpha-glucosidase inhibitor.
Compared with prior art, the present invention has following beneficial effect:
The present invention is separated from the root of Guangdong Huizhou marine site Kandelia candel mangrove and obtains a fungal strain Meyerozymasp.HZ-Y
2and separation first obtains class depsidcs 1 – 6 from the fermented liquid of this bacterial strain, it is active that these compounds have remarkable Inhibiting α-glucosidase, preparing alpha-glucosidase inhibitor medicament, has good market outlook in treatment type ii diabetes.In addition, depsidcs 1 – 6 of the present invention derives from thalassiomycetes, simple, with low cost from the method for fungi extraction and isolation.
Accompanying drawing explanation
Fig. 1 is the inhibiting two enzymatic kinetic curve reciprocal of the alpha-glucosidase of compound 1.
Fig. 2 is the inhibiting two enzymatic kinetic curve reciprocal of the alpha-glucosidase of compound 3.
Fig. 3 is the inhibiting two enzymatic kinetic curve reciprocal of the alpha-glucosidase of compound 5.
Embodiment
To make the present invention below in conjunction with Figure of description and specific embodiment and elaborating further, described embodiment, only for explaining the present invention, is not intended to limit scope of the present invention.The test method used in following embodiment if no special instructions, is ordinary method; The material used, reagent etc. if no special instructions, are the reagent that can obtain from commercial channels and material.
Embodiment 1
Thalassiomycetes Meyerozyma sp.HZ-Y of the present invention
2be be separated from the root of Guangdong Huizhou marine site Kandelia candel mangrove to obtain, separation method is with reference to this area conventional separation methods.By thalassiomycetes Meyerozyma sp.HZ-Y after separation
2be deposited in China typical culture collection center on April 6th, 2015, preservation address is Wuhan University of Wuhan, China city, and deposit number is CCTCC NO:M 2015203.
From thalassiomycetes Meyerozyma sp.HZ-Y
2fermented liquid in be separated obtain depsidcs, concrete steps are as follows:
S1. seed culture:
S11. prepare seed culture medium: potato 200g, glucose 20g, tap water 1L, average mark is loaded on 5 500mL Erlenmeyer flasks, and 121 DEG C go out 30 minutes.
S12. the cultivation of seed: by thalassiomycetes Meyerozyma sp.HZ-Y
2bacterial strain access seed culture medium, at the temperature of 28 DEG C, put with the rotating speed of 200rpm on shaking table, cultivating 72 hours must seed culture fluid.
S2. fermentation culture:
S21. prepare fermention medium: northeast rice 7000g, sea salt 210g, tap water 7L, 121 DEG C go out 30 minutes.
S22. fermentation culture: seed liquor 5mL access is equipped with in the Erlenmeyer flask of fermention medium, in 25 DEG C of quiescent culture 28 days by aseptic technique.
S3. extraction and isolation:
Fermented product methyl alcohol soaks, and soak solution concentrating under reduced pressure at lower than 50 DEG C obtains medicinal extract 12.2g.This medicinal extract is separated through silica gel column chromatography, uses 10% respectively, and 20%, 30%, 40%, 50%, the ethyl acetate-light petrol gradient elution of 60%, 70%, 80%, 100%, is divided into 30 components.Wherein the 6th component adopts dextrane gel Sephadex LH-20 chromatography, is that eluent carries out wash-out with the methanol-chloroform that volume ratio is 1:1, recycle silicon glue post layer, and 25% ethyl acetate-light petrol is eluent, obtains compound 2 and 6.8th component employing dextrane gel Sephadex LH-20 chromatography is methyl alcohol by volume ratio is that eluent carries out wash-out, and recycle silicon glue post layer, 30% ethyl acetate-light petrol is eluent, obtains compound 3 and 4.15th component adopts dextrane gel Sephadex LH-20 chromatography, is that eluent carries out wash-out, then prepares compound 1 and 5 with partly preparing RPLC with the methanol-chloroform that volume ratio is 1:1.
Embodiment 2
Structural analysis test is carried out to the compound in embodiment 1, obtains following physico-chemical property data:
Compound 1: pale yellow powder, fusing point 146-147 DEG C (thermometer does not correct), EI-MS (m/z): 288 [M]
+.
Compound 2: faint yellow needle-like crystal, fusing point 165-166 DEG C (thermometer does not correct), EI-MS (m/z): 300 [M]
+.
Compound 3: white powder, fusing point 147-148 DEG C (thermometer does not correct), EI-MS (m/z): 302 [M]
+.
Compound 4: pale yellow powder, fusing point 143-144 DEG C (thermometer does not correct), EI-MS (m/z): 316 [M]
+.
Compound 5: pale yellow oil, EI-MS (m/z): 302 [M]
+.
Compound 6: faint yellow needle-like crystal, fusing point 169-170 DEG C (thermometer does not correct), EI-MS (m/z): 314 [M]
+.
The NMR data of compound 1 – 3 are in table 1, and the NMR data of 4 – 5 are in table 2.
The NMR data (100MHz/400MHz, TMS, ppm) of table 1 compound 1 – 3
The NMR data (100MHz/400MHz, TMS, ppm) of table 2 compound 4 – 6
According to above data, learn that the structural formula of compound 1 – 6 is as shown in formula I:
Embodiment 3
Alpha-glucosidase Inhibition test is carried out to compound 1 – 6 in embodiment 1:
Adopt p-NP-alpha-glucosaccharase (pNPG) to be substrate, carry out in 0.01M phosphoric acid buffer (pH=7.0).PNPG is p-NP by alpha-glucosidase enzymolysis, to measure the change of its absorbancy and calculate the activity of enzyme with ultraviolet-visible spectrophotometer at 400nm wavelength place.Sample and positive control (acarbose) are all made into DMSO solution (being 10 μm of ol/mL), enzyme and substrate 0.01M phosphoric acid buffer are made into suitable concentration solution, 1mL initial reaction system includes 0.1unit enzyme, 20 μ L substrates, 20 μ L DMSO.Get appropriate enzyme liquid, add the DMSO solution of blank DMSO solution or sample, mixing, at 37 DEG C, constant temperature 20 minutes, adds substrate, and mixing detects the changing value of the absorbancy of system in 1min immediately at 400nm wavelength place.Enzymic activity is calculated: inhibiting rate (%)=[(B – S)/B] × 100%, wherein B is absorbancy changing value when adding blank DMSO, and S is the absorbancy changing value of sample with following formula.Measure the sample of 5 concentration, draw dosage-inhibiting rate curve, draw its IC
50value.Each sample replication three times, result mean value ± standard deviation represents.
Result records the activity of the remarkable Inhibiting α-glucosidase of compound 1 – 6 energy, its IC
50value is respectively 10.4 μMs, 13.3 μMs, 2.1 μMs, 12.4 μMs, 9.8 μMs and 11.7 μMs, the wherein IC of positive control acarbose
50it is 553.7 μMs.In addition the suppression double reciprocal curve under 3 groups of inhibitor concentration as shown in Figures 1 to 3, speed of response Vmax increases with inhibitor concentration and diminishes, and the Michaelis-Menton constant (Km) of alpha-glucosidase remains unchanged under different inhibitor concentration, double reciprocal curve meets on X-coordinate, is typical Noncompetition inhibition kinetic curve.Therefore, compound 1,3 and 5 is all noncompetitive inhibitor.
Claims (10)
1. the depsidcs in a class thalassiomycetes source, it is characterized in that, its structural formula is as shown in formula I:
Formula I depsidcs 1 – 6.
2. the preparation method of depsidcs described in claim 1, is characterized in that, depsidcs 1 – 6 is from thalassiomycetes
meyerozymain the fermented liquid of sp.HZ-Y2, separation obtains; Described thalassiomycetes
meyerozymasp.HZ-Y2 is deposited in China typical culture collection center on April 6th, 2015, and preservation address is Wuhan University of Wuhan, China city, and deposit number is CCTCC NO:M 2015203.
3. preparation method according to claim 2, is characterized in that, the preparation method of depsidcs comprises the steps:
S1. by thalassiomycetes
meyerozymasp.HZ-Y
2access seed culture medium, shaking table is cultivated, and obtains seed culture fluid;
S2. seed culture fluid is accessed in fermention medium, quiescent culture;
S3. filtered by tunning and obtain thalline, thalline methyl alcohol soaks, and concentrating under reduced pressure obtains medicinal extract, then through chromatographic separation, obtains depsidcs 1 – 6.
4. preparation method according to claim 3, is characterized in that, the component of described seed culture medium is: potato 200g, glucose 20g, water 1L.
5. preparation method according to claim 3, is characterized in that, the component of described fermention medium is: northeast rice 7000g, sea salt 210g, water 7L.
6. preparation method according to claim 3, is characterized in that, described in step S1, shaking table culture condition is: rotating speed 200rpm, temperature 28 DEG C, incubation time 72h.
7. preparation method according to claim 3, is characterized in that, quiescent culture temperature described in step S2 is 25 DEG C, and incubation time is 28 days.
8. preparation method according to claim 3, is characterized in that, described in step S3, medicinal extract silica gel column chromatography is separated, and is separated the ethyl acetate/petroleum ether gradient elution with 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 100%.
9. depsidcs described in claim 1 is preparing the application in alpha-glucosidase inhibitor.
10. the application of depsidcs according to claim 9, it is characterized in that, described alpha-glucosidase inhibitor is for preventing and treating type ii diabetes.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108640841A (en) * | 2018-06-20 | 2018-10-12 | 广东海洋大学 | A kind of depsidone class compound and its preparation method and application |
| CN108925565A (en) * | 2018-06-20 | 2018-12-04 | 广东海洋大学深圳研究院 | A kind of application of depsidone class compound |
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| WO2008144982A1 (en) * | 2007-05-29 | 2008-12-04 | Shanghai Institute Of Materia Medica, Chinese Academy Of Sciences | Compounds with 7-member cycle and the pharmaceutical use thereof for preventing and treating diabetes and metabolism syndrome |
| CN101376655A (en) * | 2008-10-11 | 2009-03-04 | 中国海洋大学 | Penicillazine derivative, and preparation and use thereof |
| EP2230235A1 (en) * | 2009-03-17 | 2010-09-22 | Leibniz-Institut für Naturstoff-Forschung und Infektionsbiologie e.V. -Hans-Knöll-Institut- (HKI) | Botryosphaerones, novel depsidones and their use as medicaments |
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2015
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2008144982A1 (en) * | 2007-05-29 | 2008-12-04 | Shanghai Institute Of Materia Medica, Chinese Academy Of Sciences | Compounds with 7-member cycle and the pharmaceutical use thereof for preventing and treating diabetes and metabolism syndrome |
| CN101376655A (en) * | 2008-10-11 | 2009-03-04 | 中国海洋大学 | Penicillazine derivative, and preparation and use thereof |
| EP2230235A1 (en) * | 2009-03-17 | 2010-09-22 | Leibniz-Institut für Naturstoff-Forschung und Infektionsbiologie e.V. -Hans-Knöll-Institut- (HKI) | Botryosphaerones, novel depsidones and their use as medicaments |
Non-Patent Citations (3)
| Title |
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| JOHN A. ELIX ET AL.: "Metabolites from the Lichen Genus Psoroma", 《AUST. J. CHEM》 * |
| JOHN A. ELIX ET AL.: "New Depsidones from the Lichen Genus Erioderma", 《AUST. J. CHEM.》 * |
| RANDA ABDOU ET AL.: "Botryorhodines A–D, antifungal and cytotoxic depsidones from Botryosphaeria rhodina, an endophyte of the medicinal plant Bidens pilosa", 《PHYTOCHEMISTRY》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108640841A (en) * | 2018-06-20 | 2018-10-12 | 广东海洋大学 | A kind of depsidone class compound and its preparation method and application |
| CN108925565A (en) * | 2018-06-20 | 2018-12-04 | 广东海洋大学深圳研究院 | A kind of application of depsidone class compound |
| CN108640841B (en) * | 2018-06-20 | 2021-04-02 | 广东海洋大学 | Depside cyclic ether compound and preparation method and application thereof |
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