CN110042132A - A kind of compound and preparation method thereof treating and preventing diabetes - Google Patents

A kind of compound and preparation method thereof treating and preventing diabetes Download PDF

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CN110042132A
CN110042132A CN201910280531.3A CN201910280531A CN110042132A CN 110042132 A CN110042132 A CN 110042132A CN 201910280531 A CN201910280531 A CN 201910280531A CN 110042132 A CN110042132 A CN 110042132A
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secondary metabolites
diabetes
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孙坤来
张译文
宫凯凯
赵成英
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Jiaxing Juetou Technology Co Ltd
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
    • A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
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    • C07D407/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having oxygen atoms as the only ring hetero atoms, not provided for by group C07D405/00
    • C07D407/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having oxygen atoms as the only ring hetero atoms, not provided for by group C07D405/00 containing three or more hetero rings
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    • 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
    • C12P17/16—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms containing two or more hetero rings
    • C12P17/162—Heterorings having oxygen atoms as the only ring heteroatoms, e.g. Lasalocid

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Abstract

The present invention provides a kind of compound and preparation method thereof for treating and preventing diabetes, belongs to microbial medicine technical field, above-mentioned secondary metabolites include that Enteromorpha source fungi isolates and purifies；Enteromorpha source fungi is inoculated in containing fermenting in TSA and enoxolone fermentation liquid；And the extraction separation and purifying of the secondary metabolites of Enteromorpha source fungi.The new application of the secondary metabolites of Enteromorpha source fungi is to prepare the new application in alpha-glucosidase restrainer and/or the drug for treating diabetes.The preparation method of secondary metabolites of the present invention can improve the yield of secondary metabolites, promote the proliferation activity of Enteromorpha source fungi, improve the yield of its secondary metabolite；Secondary metabolites of the present invention are good to the inhibitory effect of alpha-glucosidase, slow down the generation and absorption of glucose, prevent postprandial hyperglycemia, to the preferable effect of diabetes tool is prevented and treated, are easy to be absorbed by organisms.

Description

A kind of compound and preparation method thereof treating and preventing diabetes
Technical field
The invention belongs to microbial medicine technical fields, and in particular to it is a kind of treat and prevent diabetes compound and its Preparation method.
Background technique
To the research history of terrestrial microorganism last 100 years, it was found that a large amount of chemical structure diversity and bioactivity are significant Natural products, greatly pushed the development of biotin drug, such as: penicillin, vancomycin, streptomysin.To microorganism More and more concerns lead to the repetition research of microorganism and the machine of known compound overlapping development, noval chemical compound discovery naturally Rate reduces.Therefore people begin to focus on that microorganism reserves are bigger, the more marine-derived microorganisms of specific diversity.Ocean is micro- Biology is completely different with terrestrial microorganism since its unique living environment (high pressure, with high salt, low temperature, oligotrophic) causes it to have Metabolic pathway, thus be more also easy to produce the secondary metabolite different from terrestrial microorganism, and then it is living to show good biology Property such as: inhibit quorum sensing activity, bacteriostatic activity, antiviral activity, protein kinase inhibiting activity, cytotoxic activity, tumour thin Born of the same parents' cycle arresting activity etc., thus increasingly become the important natural product chemistry resource of marine drug research worker favor.Sea The abundance of microorganism reaches 10 in water6/ mL, bottom sediment bacterial abundance are even more to have reached 109/mL.However Oceanic Samples are adopted Collection difficulty is high, only isolated lower than 5% marine microorganism under typical laboratory conditions, even if the microorganism obtained exists The metabolite of its all biosynthesis pathway under marine environment can not be expressed under the conditions of laboratory cultures completely.Therefore How to develop these hard-won Marine Microorganisms to the greatest extent, can to greatest extent by human use, at For the vital task and project of scientific worker.Aspergillus fungi is all the important bacterium of natural products circle research all the time Kind, secondary metabolite structure novel skeleton is changeable, in addition to the common structures types such as conventional steroidal, sequiterpene, anthraquinone with Outside, contain toward contact: a variety of skeleton such as alkaloids, peptides, polyketone class, sesterterpene.The compound of these structure novels Containing cell toxicant, antibacterial toward contact, the various actives such as antiviral become one of the important sources of marine drug lead compound, Cause the extensive concern of scholar in the industry.
Diabetes are a kind of chronic diseases, it is characterized in that blood glucose is excessive, worldwide, diabetes oneself affected greatly There are about 3.66 hundred million people, and this number is riseing year by year, expect to be up to 5.52 hundred million to the year two thousand thirty.There are three main for diabetes Type: type 1 diabetes are also referred to as insulin-dependent diabetes mellitus, and diabetes B is referred to as non-insulin-dependent diabetes mellitus and pregnant It is pregnent phase diabetes.Diabetes B is the most common form, and the patient in such case is more than 90%.Except through diet and Movement is to control other than blood glucose level, and there are also some remedy measures to control blood glucose level in a certain range, mainly at present It is 6 major class drugs, including insulin of oneself approved listing and the like, sulfonylureas, meglitinides, biguanides, thiazole Alkane diones (TZDs) and alpha-glucosidase inhibitor.And alpha-glucosidase inhibitor has toward antidiabetic drug institute not as a kind of antidiabetic drug It has the advantage that, and such drug of clinical use only has few in number several at present, therefore alpha-glucosidase inhibitor is opened Hair has boundless prospect, while being also the important directions of relationship human health.
Summary of the invention
The purpose of the present invention is to provide a kind of versatility related gene expressions for promoting Enteromorpha source fungi, improve secondary The yield of metabolin, while promoting the proliferation activity of Enteromorpha source fungi, the yield of its secondary metabolite is improved, is microorganism Field of medicaments provides the preparation method of the compound of the treatment and prevention diabetes of new approach.
The technical solution that the present invention is taken to achieve the above object are as follows:
A kind of preparation method for the compound treating and preventing diabetes, including,
Enteromorpha source fungi isolates and purifies；
Above-mentioned Enteromorpha source fungi is inoculated in containing fermenting in TSA and enoxolone fermentation liquid；And
The extraction of the secondary metabolites of Enteromorpha source fungi separates and purifying.
TSA and enoxolone can generate the facilitation of collaboration to the fermentation of Enteromorpha source fungi in fermentation liquid, not Same cell-signaling pathways effect is lower to carry out cytoplasm and nucleus shuttle, becomes slack high degree of coagulation Chromatin domains, removal Acetyl group on histone ε-N-acetyllysine residue, so that DNA is more closely wrapped on histone, and can be with it His transcription factor regulatory protein forms co-suppression compound, and enhancing and the amino acid residue at enzyme active sites edge interact, Inhibitory effect and the selectivity to enzyme are improved, so that inhibition of gene expression, enhances the degree of acetylation of histone, transcription is promoted to swash It is living, cause transcriptional level to increase, the selective expression of controlling gene, promotes in the case where not changing genomic dna sequence The versatility related gene expression of Enteromorpha source fungi improves the yield of secondary metabolites, while it is true to regulate and control Enteromorpha source The expression of bacterium nitrogen metabolism related gene, adjusts the level of intracellular glutamine synthelase, promote fungi to the absorption of nitrogen source and It utilizes, and then promotes the proliferation activity of Enteromorpha source fungi, improve the yield of its secondary metabolite, be microorganism field of medicaments New approach is provided.
Preferably, Enteromorpha source fungi is Aspergillus terreus OUCMDZ-2739.
Preferably, 9-11 μM of TSA and 0.8-1.3 μM of enoxolone are contained in fermentation liquid.
Preferably, the purification step of the secondary metabolites of Enteromorpha source fungi are as follows: ethyl acetate extract is utilized into decompression Silica gel column chromatography, Sephadex LH-20, HPLC are further purified, and obtain target compound
It is furthermore preferred that HPLC, which is used, contains 60%MeOH-H2The solution of O and 0.15%TFA elutes.
It is furthermore preferred that the TR of secondary metabolites is 17.6 minutes during HPLC.
It is good that another object of the present invention is to provide the inhibitory effects of a kind of pair of alpha-glucosidase, slows down the life of glucose At and absorb, prevent postprandial hyperglycemia, to the preferable effect of diabetes tool is prevented and treated, be easy to be absorbed by organisms is controlled Treat and prevent the compound of diabetes.
The technical solution that the present invention is taken to achieve the above object are as follows:
Compound made from a kind of preparation method of the compound of above-mentioned treatment and prevention diabetes, structural formula are as follows:
Half-inhibitory concentration (IC of the above compound to alpha-glucosidase50) value be 24.8 μM.1-Deoxynojirimycin and The IC of acarbose50Value is respectively 191.7 μM and 555.1 μM, illustrates that compound 8 is better than the inhibitory effect of alpha-glucosidase 1-Deoxynojirimycin and acarbose, this shows that reactive compound is better than acarbose to the inhibitory effect of alpha-glucosidase, Inhibit body alpha-glucosidase activity, slow down the generation and absorption of glucose, prevent postprandial hyperglycemia, to preventing and control Treat the preferable effect of diabetes tool, especially diabetes B.
Above-mentioned diabetes are type 1 diabetes or diabetes B.
Compared with prior art, the invention has the benefit that
The preparation method of the compounds of this invention promotes the versatility related gene expression of Enteromorpha source fungi, improves secondary generation It thanks to the yield of object, while promoting the proliferation activity of Enteromorpha source fungi, improve the yield of its secondary metabolite, cured for microorganism Medicine field provides new approach；Half-inhibitory concentration (the IC of the compounds of this invention alpha-glucosidase50) value be 24.8 μM, to α- The inhibitory effect of glucuroide is better than 1-Deoxynojirimycin and acarbose, and the inhibitory effect of alpha-glucosidase is better than Ah Ka Wave sugar, inhibits body alpha-glucosidase activity, slows down the generation and absorption of glucose, postprandial hyperglycemia is prevented, to pre- Anti- and the preferable effect for the treatment of diabetes tool, especially diabetes B；Secondary of the compounds of this invention from microorganism simultaneously Metabolite belongs to natural alpha-glucosidase restrainer, is easy to be absorbed by organisms.
Present invention employs above-mentioned technical proposals to provide a kind of compound and preparation method thereof for treating and preventing diabetes, Compensate for the deficiencies in the prior art, reasonable design, easy operation.
Detailed description of the invention
Fig. 1 is the ECD curve of the compounds of this invention.
Specific embodiment
The application discloses a kind of preparation method of compound for treating and preventing diabetes, including,
Enteromorpha source fungi isolates and purifies；
Above-mentioned Enteromorpha source fungi is inoculated in containing fermenting in TSA and enoxolone fermentation liquid；And
The extraction of the secondary metabolites of Enteromorpha source fungi separates and purifying.
TSA and enoxolone can generate the rush of collaboration to the fermentation of Enteromorpha source fungi in the preparation method fermentation liquid Into effect, cytoplasm is carried out under different cell-signaling pathways effects and nucleus shuttles, makes high degree of coagulation Chromatin domains It becomes slack, the acetyl group on histone ε-N-acetyllysine residue is removed, so that DNA is more closely wrapped in histone On, and co-suppression compound, the amino acid of enhancing and enzyme active sites edge can be formed with other transcription factor regulatory proteins Residue interaction, improves inhibitory effect and the selectivity to enzyme, so that inhibition of gene expression, enhances the acetylation journey of histone Degree promotes transcriptional activation, causes transcriptional level to increase, the selection of controlling gene in the case where not changing genomic dna sequence Property expression, promote the versatility related gene expression of Enteromorpha source fungi, improve the yield of secondary metabolites, while can adjust The expression for controlling Enteromorpha source fungi nitrogen metabolism related gene adjusts the level of intracellular glutamine synthelase, promotes fungi pair The absorption and utilization of nitrogen source, and then promote the proliferation activity of Enteromorpha source fungi, the yield of its secondary metabolite is improved, is micro- Biomedicine field provides new approach, and furthermore gained target compound accounting in ethyl acetate solution extract increases Add, further explains the facilitation that TSA and enoxolone in fermentation liquid generate collaboration to the fermentation of Enteromorpha source fungi.
Above-mentioned Enteromorpha source fungi is Aspergillus terreus OUCMDZ-2739.
Contain 9-11 μM of TSA (such as 9.2 μM, 9.5 μM, 10.3 μM, 10.8 μM etc.) and 0.8-1.3 μ in above-mentioned fermentation liquid M enoxolone (such as 0.83 μM, 0.95 μM, 1.0 μM, 1.1 μM, 1.2 μM, 1.27 μM etc.).
The purification step of the secondary metabolites of above-mentioned Enteromorpha source fungi are as follows: ethyl acetate extract is utilized into decompression silica gel Column chromatography, Sephadex LH-20, HPLC are further purified, and obtain target compound
Above-mentioned HPLC, which is used, contains 60%MeOH-H2The solution of O and 0.15%TFA elutes.
The TR of secondary metabolites is 17.6 minutes during above-mentioned HPLC.
Compound made from the preparation method of the compound of above-mentioned treatment and prevention diabetes, structure is also disclosed in the application Formula are as follows:
Half-inhibitory concentration (IC of the above compound to alpha-glucosidase50) value be 24.8 μM.1-Deoxynojirimycin and The IC of acarbose50Value is respectively 191.7 μM and 555.1 μM, illustrates that compound 8 is better than the inhibitory effect of alpha-glucosidase 1-Deoxynojirimycin and acarbose, this shows that reactive compound is better than acarbose to the inhibitory effect of alpha-glucosidase, Inhibit body alpha-glucosidase activity, slow down the generation and absorption of glucose, prevent postprandial hyperglycemia, to preventing and control Treat the preferable effect of diabetes tool, especially diabetes B.
Above-mentioned diabetes are type 1 diabetes or diabetes B.
In the following, being described further in conjunction with specific embodiments to embodiment of the present invention.
Embodiment 1:
A kind of preparation method for the compound treating and preventing diabetes, including,
1) Enteromorpha source fungi isolates and purifies: isolating fungi from Qingdao Stone old man bathing beach Enteromorpha sample Enteromorpha sample is successively used antiseptic sea water, 75% ethyl alcohol and sterile water washing by bacterial strain Aspergillus terreus OUCMDZ-2739, then, with grinding Alms bowl smashs sample to pieces, then deposits in PDA culture medium (every liter of 200g containing potato extract, Portugal containing 100mg/mL chloramphenicol Grape sugar 20g, agar 15g and seawater 1L) in, it is cultivated at 28 DEG C to there is single culture, which is transferred to another PDA Culture medium is simultaneously stored in 4 DEG C, passes through polyphase sort research, colonial morphology and the analysis of 18S rRNA sequence, strain development tree It establishes, is identified as Aspergillus terreus OUCMDZ-2739；
2) Enteromorpha source fungi is inoculated in containing fermenting 30 days at 25 DEG C in TSA and enoxolone fermentation liquid, fermentation liquid By glucose (10g/L), maltose (20g/L), mannitol (20g/L), monosodium glutamate (10g/L), KH2PO4(0.5g/L)、 MgSO4·7H2O (0.3g/L), yeast extract (3g/L), SEA salt (33g/L), TSA (10 μM) and tap water (1L, pH 7) group At；
3) the extraction separation and purifying of the secondary metabolites of Enteromorpha source fungi: fermentation liquid is filtered by garrha to divide From filtrate and mycelia, the ethyl acetate extraction of filtrate equivalent volume obtains ethyl acetate solution, 80% acetone of mycelia three times Three times, then acetone soln obtains ethyl acetate solution through being concentrated under reduced pressure with ethyl acetate extraction three times for extraction, will be above-mentioned two Ethyl acetate solution is combined, and is concentrated under reduced pressure, and 31.2g ethyl acetate solution extract is obtained；Then by acetic acid Ethyl ester extract is gradually eluted using decompression silica gel column chromatography, and eluent is petroleum ether-CH2Cl2(1:1 and 0:1) and CH2Cl26 main fractions of surrender gradient (fraction 1~6) of-MeOH (100%~0%), the fraction 4 of 5.2g pass through Sephadex LH-20 (MeOH) is further purified, and obtains 260mg fraction 4.2,120mg fraction 4.3 and 370mg fraction 4.4, wherein fraction 4.2 Continue to be purified with HPLC, eluent 60%MeOH-H2O and 0.15%TFA obtains 12.1mg target compound (tR= 17.6min)。
Embodiment 2:
A kind of preparation method for the compound treating and preventing diabetes, including,
1) Enteromorpha source fungi isolates and purifies: isolating fungi from Qingdao Stone old man bathing beach Enteromorpha sample Enteromorpha sample is successively used antiseptic sea water, 75% ethyl alcohol and sterile water washing by bacterial strain Aspergillus terreus OUCMDZ-2739, then, with grinding Alms bowl smashs sample to pieces, then deposits in PDA culture medium (every liter of 200g containing potato extract, Portugal containing 100mg/mL chloramphenicol Grape sugar 20g, agar 15g and seawater 1L) in, it is cultivated at 28 DEG C to there is single culture, which is transferred to another PDA Culture medium is simultaneously stored in 4 DEG C, passes through polyphase sort research, colonial morphology and the analysis of 18S rRNA sequence, strain development tree It establishes, is identified as Aspergillus terreus OUCMDZ-2739；
2) Enteromorpha source fungi is inoculated in containing fermenting 30 days at 25 DEG C in TSA and enoxolone fermentation liquid, fermentation liquid By glucose (10g/L), maltose (20g/L), mannitol (20g/L), monosodium glutamate (10g/L), KH2PO4(0.5g/L)、 MgSO4·7H2O (0.3g/L), yeast extract (3g/L), SEA salt (33g/L), TSA (10 μM), enoxolone (1.0 μM) and Tap water (1L, pH 7) composition；
3) the extraction separation and purifying of the secondary metabolites of Enteromorpha source fungi: fermentation liquid is filtered by garrha to divide From filtrate and mycelia, the ethyl acetate extraction of filtrate equivalent volume obtains ethyl acetate solution, 80% acetone of mycelia three times Three times, then acetone soln obtains ethyl acetate solution through being concentrated under reduced pressure with ethyl acetate extraction three times for extraction, will be above-mentioned two Ethyl acetate solution is combined, and is concentrated under reduced pressure, and 37.5g ethyl acetate solution extract is obtained；Then by acetic acid Ethyl ester extract is gradually eluted using decompression silica gel column chromatography, and eluent is petroleum ether-CH2Cl2(1:1 and 0:1) and CH2Cl26 main fractions of surrender gradient (fraction 1~6) of-MeOH (100%~0%), the fraction 4 of 7.2g pass through Sephadex LH-20 (MeOH) is further purified, and obtains 378.1mg fraction 4.2,168.5mg fraction 4.3 and 517.1mg fraction 4.4, wherein Fraction 4.2 continues to be purified with HPLC, eluent 60%MeOH-H2O and 0.15%TFA obtains 17.7mg target compound (tR =17.6min).
Compound made from a kind of preparation method of the compound of above-mentioned treatment and prevention diabetes, structural formula are as follows:
Above compound is yellowish solid, [a] D25-36(c 0.1,CH2Cl2)；UV(MeOH)λmax(log ε): 204 (4.35),252(3.95)and 376(4.25)nm；ECD(0.0022M,MeOH)λmax(△ε)207(+1.13),230(- 0.13), 252 (+0.10), 373 (- 0.63) nm (such as Fig. 1)；IR(KBr)νmax 3747,3641,1696,1536,1501cm-1； 1H and 13C NMR value such as table 1；((calcd.for C27H28O6, 449.1959).
1 compound of table is in CDCl3In1H(500MHz)and 13C (125MHz) NMR value
Position δH(J/Hz) δC
1 167.6,s
2 106.0,s
3 157.3,s
4 142.0,s
5 6.73,s 112.9,d
1′ 120.1,s
2′ 7.66,s 129.2,d
3′ 121.2,s
4′ 154.2,s
5′ 6.80,d(8.6) 117.5,d
6′ 7.62,d(8.5) 127.1,d
1″ 123.9,s
2″ 7.31,s 125.9,d
3″ 129.4,s
4″ 160.6,s
5″ 6.86,d(8.2) 110.2,d
6″ 7.25,d(8.1) 129.1,d
1″′ 2.79,t(6.6) 22.5,t
2′ 1.80,t(6.7) 32.7,t
3″′ 74.7,s
4″′ 1.33,s 26.9,q
5″′ 1.33,s 26.9,q
Embodiment 3:
In order to improve the separating degree of target compound, further prioritization scheme are as follows:
Using containing 60%MeOH-H in HPLC purification process2O, the solution of 0.35 ‰ phenyl-phosphonic acids and 0.15%TFA elute. MeOH (methanol) is used as intensive polar solvent, can be dissolved in just own burning/awns alcohol or just own burning/aqueous isopropanol, and polarity is big and proton Property it is strong, it is cheap, purification be easy, TFA (trifluoroacetic acid) is the good solvent of many organic compounds, the trifluoro in mobile phase For acetic acid by interacting in different modes with hydrophobic bonding phase and remaining polar surfaces, Lai Gaishan peak shape overcomes peak stretching And tailing problem；It can be protonated in target compound, the MeOH, phenyl-phosphonic acid and TFA in above-mentioned solution can play gain Effect can provide sufficient proton, can be with so that exchange is more fully adsorbed and desorbed to target compound and stationary phase , there is independent chromatographic peak, while causing stationary phase and target compound in the appearance time for shortening target compound in chromatogram Between hydrogen bond action increase so that separating degree and separation factor increase.In contrast, add 0.35 ‰ phenyl-phosphonic acids and Under conditions of 0.15%TFA, the separating degree of each chromatographic peak is best, and appearance time is most short, so 0.35 ‰ phenyl-phosphonic acids of selection With 0.15%TFA as most suitable additive amount, 19.1mg target compound is obtained.
Embodiment 4:
Target compound tests the inhibition of alpha-glucosidase:
The preparation of alpha-glucosaccharase enzyme solutions: enzyme freeze-dried powder 0.1%BSA solution is dissolved, the enzyme of 100U/mL is made into Liquid is placed in -20 DEG C of refrigerator and freezes.Before experiment, 100U/mL enzyme solution liquid-transfering gun is drawn on a small quantity, it is first molten with 0.1%BSA Liquid is diluted to 20U/mL, then with 0.1%BSA solution to be diluted to 0.2U/mL spare.
PNPG solution is prepared: weighing a certain amount of PNPG solid, the phosphate buffer for being 6.8 with 0.1mol/L pH (PBS) it dissolves, being made into concentration is 10mmol/L, then spare with the centrifuge tube packing of 1.5mL.
Using the activity of colorimetric method for determining alpha-glucosidase.Active ingredient to be measured is dissolved with dimethyl sulfoxide (DMSO) Object prepares the stock solution that mass concentration is 10mg/mL.Stock solution is diluted with PBS again, is configured to the to be measured molten of required concentration Liquid.Active compounds solution (experimental group), acarbose solution (positive controls) or PBS (yin are added in the every hole of 96 orifice plates Property control group) 10 μ L, 50 μ L of PBS, 20 μ L of enzyme solutions, shake up, be placed in 37 DEG C of constant temperature water bath 10min, then be added PNPG solution 20 μ L, shake up, and 37 DEG C of reaction 10min add Na2CO330 μ L of terminate liquid terminates reaction, measures absorbance at 405nm immediately Value.The inhibiting rate of alpha-glucosidase is calculated as follows in sample: inhibiting rate (%)=[(negative control group absorbance-experiment Group absorbance)/negative control group absorbance] × 100%.When inhibiting rate of the sample to alpha-glucosidase is 50%, by sample Quality concentration is set to half-inhibitory concentration (IC50) value.Target compound is measured to the half-inhibitory concentration of alpha-glucosidase (IC50) value be 24.8 μM.The IC of 1-Deoxynojirimycin and acarbose50Value is respectively 191.7 μM and 555.1 μM, illustrates target Compound is better than 1-Deoxynojirimycin and acarbose to the inhibitory effect of alpha-glucosidase.And above-mentioned secondary metabolites pair Ki value is 1.42 μM.This shows that reactive compound is better than acarbose to the inhibitory effect of alpha-glucosidase, inhibits body α- Glucosidase activity slows down the generation and absorption of glucose, prevents postprandial hyperglycemia, has to diabetes are prevented and treated Preferable effect, especially diabetes B.
The prior art of routine techniques dawn known to those skilled in the art in above-described embodiment, therefore herein no longer in detail It repeats.
The above embodiments are only used to illustrate the present invention, and not limitation of the present invention, the ordinary skill people of this field Member can also make a variety of changes and modification without departing from the spirit and scope of the present invention.Therefore, all equivalent Technical solution also belong to scope of the invention, scope of patent protection of the invention should be defined by the claims.

Claims (10)

1. a kind of preparation method for the compound for treating and preventing diabetes, it is characterised in that: including point of Enteromorpha source fungi From purifying；
Enteromorpha source fungi is inoculated in containing fermenting in TSA and enoxolone fermentation liquid；And
The extraction of the secondary metabolites of Enteromorpha source fungi separates and purifying.
2. a kind of preparation method of compound for treating and preventing diabetes according to claim 1, it is characterised in that: institute Stating Enteromorpha source fungi is Aspergillus terreus OUCMDZ-2739.
3. a kind of preparation method of compound for treating and preventing diabetes according to claim 1, it is characterised in that: institute It states and contains 9-11 μM of TSA and 0.8-1.3 μM of enoxolone in fermentation liquid.
4. a kind of preparation method of the secondary metabolites of Enteromorpha source fungi according to claim 1, it is characterised in that: institute State the extraction separating step of the secondary metabolites of Enteromorpha source fungi are as follows: fermentation liquid is filtered by garrha with separating filtrate and The ethyl acetate extraction of mycelia, filtrate equivalent volume obtains ethyl acetate solution, 80% acetone of the mycelia three times Three times, then acetone soln obtains ethyl acetate solution through being concentrated under reduced pressure with ethyl acetate extraction three times for extraction, will be described two Ethyl acetate solution is combined, and is concentrated under reduced pressure, and ethyl acetate solution extract is obtained.
5. a kind of preparation method of compound for treating and preventing diabetes according to claim 1, it is characterised in that: institute State the purification step of the secondary metabolites of Enteromorpha source fungi are as follows: the ethyl acetate extract is utilized into decompression silicagel column color Spectrum, Sephadex LH-20, HPLC are further purified, and obtain target compound.
6. a kind of preparation method of compound for treating and preventing diabetes according to claim 5, it is characterised in that: institute HPLC is stated to use containing 60%MeOH-H2The solution of O and 0.15%TFA elutes.
7. a kind of preparation method of compound for treating and preventing diabetes according to claim 5, it is characterised in that: institute The TR for stating secondary metabolites during HPLC is 17.6 minutes.
8. chemical combination made from a kind of preparation method of any one of claim 1-5 compound for treating and preventing diabetes Object, it is characterised in that: structural formula are as follows:
9. a kind of compound for treating and preventing diabetes according to claim 7, it is characterised in that: the compound pair Half-inhibitory concentration (the IC of alpha-glucosidase50) value be 24.8 μM.
10. a kind of compound for treating and preventing diabetes according to claim 7, it is characterised in that: the diabetes For type 1 diabetes or diabetes B.
CN201910280531.3A 2019-04-09 2019-04-09 A kind of compound and preparation method thereof treating and preventing diabetes Withdrawn CN110042132A (en)

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Cited By (3)

* Cited by examiner, † Cited by third party
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CN115154453A (en) * 2021-08-17 2022-10-11 周口师范学院 Application of aspulvinone compound in preparation of antidiabetic drugs
CN115894467A (en) * 2021-08-17 2023-04-04 周口师范学院 Dicyclo diisopentenyl substituted aspulvinone compound and preparation method and application thereof
CN115991687A (en) * 2021-08-17 2023-04-21 周口师范学院 Diisopentenyl substituted aspulvinone compound and preparation method and application thereof

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN115154453A (en) * 2021-08-17 2022-10-11 周口师范学院 Application of aspulvinone compound in preparation of antidiabetic drugs
CN115894467A (en) * 2021-08-17 2023-04-04 周口师范学院 Dicyclo diisopentenyl substituted aspulvinone compound and preparation method and application thereof
CN115991687A (en) * 2021-08-17 2023-04-21 周口师范学院 Diisopentenyl substituted aspulvinone compound and preparation method and application thereof
CN115154453B (en) * 2021-08-17 2024-01-19 周口师范学院 Application of aspulvinone compound in preparation of antidiabetic drugs
CN115894467B (en) * 2021-08-17 2024-05-10 周口师范学院 Dicyclo-diisopentenyl substituted aspulvinone compound and preparation method and application thereof
CN115991687B (en) * 2021-08-17 2024-09-20 周口师范学院 Diisopentenyl substituted aspulvinone compound and preparation method and application thereof

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