NL2033900B1 - Monascuskaolin a, b and c having glycosidase inhibitory, immunoregulatory and anti-aging activities and preparation method thereof - Google Patents
Monascuskaolin a, b and c having glycosidase inhibitory, immunoregulatory and anti-aging activities and preparation method thereof Download PDFInfo
- Publication number
- NL2033900B1 NL2033900B1 NL2033900A NL2033900A NL2033900B1 NL 2033900 B1 NL2033900 B1 NL 2033900B1 NL 2033900 A NL2033900 A NL 2033900A NL 2033900 A NL2033900 A NL 2033900A NL 2033900 B1 NL2033900 B1 NL 2033900B1
- Authority
- NL
- Netherlands
- Prior art keywords
- monascus
- monascuskaolin
- glycosidase
- kaolin
- tgf
- Prior art date
Links
- 108010031186 Glycoside Hydrolases Proteins 0.000 title claims abstract description 24
- 102000005744 Glycoside Hydrolases Human genes 0.000 title claims abstract description 24
- 230000002401 inhibitory effect Effects 0.000 title claims abstract description 20
- 230000003712 anti-aging effect Effects 0.000 title claims abstract description 15
- 230000004957 immunoregulator effect Effects 0.000 title claims abstract description 9
- 238000002360 preparation method Methods 0.000 title abstract description 20
- 241000255588 Tephritidae Species 0.000 claims abstract description 10
- 238000000338 in vitro Methods 0.000 claims abstract description 10
- 230000005764 inhibitory process Effects 0.000 claims abstract description 10
- 210000002540 macrophage Anatomy 0.000 claims abstract description 10
- 239000000843 powder Substances 0.000 claims abstract description 9
- 108060008682 Tumor Necrosis Factor Proteins 0.000 claims abstract description 7
- 102000004887 Transforming Growth Factor beta Human genes 0.000 claims abstract 3
- 108090001012 Transforming Growth Factor beta Proteins 0.000 claims abstract 3
- ZRKFYGHZFMAOKI-QMGMOQQFSA-N tgfbeta Chemical compound C([C@H](NC(=O)[C@H](C(C)C)NC(=O)CNC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CC(C)C)NC(=O)CNC(=O)[C@H](C)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](NC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)[C@@H](NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CCSC)C(C)C)[C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](C)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(=O)N1[C@@H](CCC1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O)C1=CC=C(O)C=C1 ZRKFYGHZFMAOKI-QMGMOQQFSA-N 0.000 claims abstract 3
- MZOFCQQQCNRIBI-VMXHOPILSA-N (3s)-4-[[(2s)-1-[[(2s)-1-[[(1s)-1-carboxy-2-hydroxyethyl]amino]-4-methyl-1-oxopentan-2-yl]amino]-5-(diaminomethylideneamino)-1-oxopentan-2-yl]amino]-3-[[2-[[(2s)-2,6-diaminohexanoyl]amino]acetyl]amino]-4-oxobutanoic acid Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC(O)=O)NC(=O)CNC(=O)[C@@H](N)CCCCN MZOFCQQQCNRIBI-VMXHOPILSA-N 0.000 claims abstract 2
- 102000000852 Tumor Necrosis Factor-alpha Human genes 0.000 claims abstract 2
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims description 27
- 241001095209 Monascus sp. (in: Fungi) Species 0.000 claims description 14
- 241000228347 Monascus <ascomycete fungus> Species 0.000 claims description 13
- 102000000589 Interleukin-1 Human genes 0.000 claims description 4
- 108010002352 Interleukin-1 Proteins 0.000 claims description 4
- 239000005995 Aluminium silicate Substances 0.000 claims 12
- 235000012211 aluminium silicate Nutrition 0.000 claims 12
- NLYAJNPCOHFWQQ-UHFFFAOYSA-N kaolin Chemical compound O.O.O=[Al]O[Si](=O)O[Si](=O)O[Al]=O NLYAJNPCOHFWQQ-UHFFFAOYSA-N 0.000 claims 12
- 230000035897 transcription Effects 0.000 claims 3
- 238000013518 transcription Methods 0.000 claims 3
- 230000003110 anti-inflammatory effect Effects 0.000 abstract description 2
- 238000010924 continuous production Methods 0.000 abstract description 2
- 239000003316 glycosidase inhibitor Substances 0.000 abstract 1
- 229960003444 immunosuppressant agent Drugs 0.000 abstract 1
- 239000003018 immunosuppressive agent Substances 0.000 abstract 1
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 9
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 9
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 9
- 238000012258 culturing Methods 0.000 description 8
- 238000009630 liquid culture Methods 0.000 description 8
- 239000007788 liquid Substances 0.000 description 7
- 239000003480 eluent Substances 0.000 description 6
- 238000000605 extraction Methods 0.000 description 6
- 102100040247 Tumor necrosis factor Human genes 0.000 description 5
- 239000001963 growth medium Substances 0.000 description 5
- 239000001965 potato dextrose agar Substances 0.000 description 5
- 238000007670 refining Methods 0.000 description 5
- 244000113306 Monascus purpureus Species 0.000 description 4
- 235000002322 Monascus purpureus Nutrition 0.000 description 4
- 150000001875 compounds Chemical class 0.000 description 4
- 239000012043 crude product Substances 0.000 description 4
- 239000003814 drug Substances 0.000 description 4
- 229940079593 drug Drugs 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 235000013305 food Nutrition 0.000 description 4
- 239000000047 product Substances 0.000 description 4
- 239000000725 suspension Substances 0.000 description 4
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 3
- 238000005160 1H NMR spectroscopy Methods 0.000 description 3
- XILIYVSXLSWUAI-UHFFFAOYSA-N 2-(diethylamino)ethyl n'-phenylcarbamimidothioate;dihydrobromide Chemical compound Br.Br.CCN(CC)CCSC(N)=NC1=CC=CC=C1 XILIYVSXLSWUAI-UHFFFAOYSA-N 0.000 description 3
- 238000005481 NMR spectroscopy Methods 0.000 description 3
- 239000003472 antidiabetic agent Substances 0.000 description 3
- 150000001768 cations Chemical class 0.000 description 3
- 238000005100 correlation spectroscopy Methods 0.000 description 3
- 206010012601 diabetes mellitus Diseases 0.000 description 3
- 229940093499 ethyl acetate Drugs 0.000 description 3
- 235000019439 ethyl acetate Nutrition 0.000 description 3
- 238000001704 evaporation Methods 0.000 description 3
- 230000008020 evaporation Effects 0.000 description 3
- 238000003919 heteronuclear multiple bond coherence Methods 0.000 description 3
- 238000003929 heteronuclear multiple quantum coherence Methods 0.000 description 3
- 238000004895 liquid chromatography mass spectrometry Methods 0.000 description 3
- 238000000746 purification Methods 0.000 description 3
- 239000012264 purified product Substances 0.000 description 3
- 238000004611 spectroscopical analysis Methods 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 2
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 2
- 241000031003 Monascus ruber Species 0.000 description 2
- 235000002595 Solanum tuberosum Nutrition 0.000 description 2
- 244000061456 Solanum tuberosum Species 0.000 description 2
- 230000003321 amplification Effects 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- 230000001413 cellular effect Effects 0.000 description 2
- 238000013375 chromatographic separation Methods 0.000 description 2
- 230000006837 decompression Effects 0.000 description 2
- 238000010828 elution Methods 0.000 description 2
- 239000000706 filtrate Substances 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- 229910052739 hydrogen Inorganic materials 0.000 description 2
- 239000001257 hydrogen Substances 0.000 description 2
- 230000002218 hypoglycaemic effect Effects 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- 238000011081 inoculation Methods 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 238000000034 method Methods 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 229940057059 monascus purpureus Drugs 0.000 description 2
- 238000003199 nucleic acid amplification method Methods 0.000 description 2
- 239000002244 precipitate Substances 0.000 description 2
- 238000004366 reverse phase liquid chromatography Methods 0.000 description 2
- 238000004007 reversed phase HPLC Methods 0.000 description 2
- 238000012807 shake-flask culturing Methods 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 229910021642 ultra pure water Inorganic materials 0.000 description 2
- 239000012498 ultrapure water Substances 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 238000005084 2D-nuclear magnetic resonance Methods 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 241001326554 Elaphomycetaceae Species 0.000 description 1
- 241000221785 Erysiphales Species 0.000 description 1
- 108090000371 Esterases Proteins 0.000 description 1
- 241000228427 Eurotiales Species 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 108010073178 Glucan 1,4-alpha-Glucosidase Proteins 0.000 description 1
- 102100022624 Glucoamylase Human genes 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 150000001298 alcohols Chemical class 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 235000021107 fermented food Nutrition 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 235000019634 flavors Nutrition 0.000 description 1
- 230000005965 immune activity Effects 0.000 description 1
- 208000026278 immune system disease Diseases 0.000 description 1
- 230000001506 immunosuppresive effect Effects 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 238000004949 mass spectrometry Methods 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- PSGAAPLEWMOORI-PEINSRQWSA-N medroxyprogesterone acetate Chemical compound C([C@@]12C)CC(=O)C=C1[C@@H](C)C[C@@H]1[C@@H]2CC[C@]2(C)[C@@](OC(C)=O)(C(C)=O)CC[C@H]21 PSGAAPLEWMOORI-PEINSRQWSA-N 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 150000002772 monosaccharides Chemical class 0.000 description 1
- 239000002773 nucleotide Substances 0.000 description 1
- 125000003729 nucleotide group Chemical group 0.000 description 1
- 238000000238 one-dimensional nuclear magnetic resonance spectroscopy Methods 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 239000008223 sterile water Substances 0.000 description 1
- 230000002195 synergetic effect Effects 0.000 description 1
- 238000002137 ultrasound extraction Methods 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
- A61K31/365—Lactones
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
-
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P39/00—General protective or antinoxious agents
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D493/00—Heterocyclic compounds containing oxygen atoms as the only ring hetero atoms in the condensed system
- C07D493/02—Heterocyclic compounds containing oxygen atoms as the only ring hetero atoms in the condensed system in which the condensed system contains two hetero rings
- C07D493/04—Ortho-condensed systems
-
- 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
- C12P17/18—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms containing at least two hetero rings condensed among themselves or condensed with a common carbocyclic ring system, e.g. rifamycin
- C12P17/181—Heterocyclic compounds containing oxygen atoms as the only ring heteroatoms in the condensed system, e.g. Salinomycin, Septamycin
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/645—Fungi ; Processes using fungi
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Diabetes (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Immunology (AREA)
- Pain & Pain Management (AREA)
- Zoology (AREA)
- Emergency Medicine (AREA)
- Endocrinology (AREA)
- Hematology (AREA)
- Obesity (AREA)
- Epidemiology (AREA)
- Rheumatology (AREA)
- Wood Science & Technology (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Genetics & Genomics (AREA)
- Toxicology (AREA)
- Biotechnology (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The present invention relates to Monascuskaolin A, B and C having glycosidase inhibitory, immunoregulatory and anti—aging activities and a preparation method thereof. Monascuskaolin A, B and C are reddish—brown powder, with leecular formulas of CEHQO7, CBHwO6 and, Clflh205; the Monascuskaolin. A, B and, C have a glycosidase inhibitory activity, and, median inhibition concentrations (ICw) thereof against alpha—glycosidase are 220 micrograms/mL, 174 micrograms/mL and 200 micrograms/mL respectively, and the Monascuskaolin. A, B and, C can significantly downregulate transcript levels of IL—l beta, TNF—alpha and TGF—beta of in vitro cultured mouse macrophages RAW264.7; and the life spans of fruit flies are extended by 31.6 percent, 27.3 percent and 22.5 percent, respectively. The Monascuskaolin A, B and C can be used for preparing glycosidase inhibitors, anti—inflammatory immunosuppressants and anti—aging preparations. The preparation method is environmental—friendly, is not affected by natural environment and, resources, and, is liable to realize industrial, automatic and continuous production.
Description
MONASCUSKAOLIN A, B AND C HAVING GLYCOSIDASE INHIBITORY,
IMMUNOREGULATORY AND ANTI-AGING ACTIVITIES AND PREPARATION METHOD
THEREOF
The present invention belongs to the technical field of ex- traction and preparation of biological products, and particularly relates to Monascuskaolin A, B and C having glycosidase inhibito- ry, immunoregulatory and anti-aging activities, as well as extrac- tion, purification, structural identification and activity deter- mination thereof.
Monascus sp., a kind of small-sized saprophytic fungi of
Monascus in Monascaceae, Eurotiales, Plectomycetes, Ascomycotina,
Eumycophyta, includes Monascus purpureus, Monascus anka and Monas- cus ruber. In China and Southeast Asia, Monascus sp. has been ap- plied in food for more than 1,000 years. Monascus sp. products are important food pigments and can produce enzyme substances such as glucoamylase, esterase and protease. Under the synergistic effect of these enzymes, taste compounds such as monosaccharide, amino acids and nucleotides, and aroma compounds such as volatile acids, alcohols and esters can be produced, contributing significantly to the flavor of fermented foods.
In recent years, the number of diabetic patients has in- creased year by year due to improved living standards and reduced amount of exercise. At present, there are 440 million diabetic pa- tients in the world and the figure increases by 5% per year. How- ever, there are few kinds and options of hypoglycemic drugs, so it is urgent to develop new and efficient hypoglycemic drugs. Devel- oping relevant drugs can bring great social and economic benefits.
With the rapid development of today’s society, due to the fast pace of life and work and great stress, lack of exercise, staying up late to check mobile phones and other bad habits, the number of sub-healthy people with immune disorders increases year by year. Therefore, it is necessary to develop immunoregulatory drugs. As China and many countries around the world have entered into the aged society, anti-aging products are in short supply. As a food microorganism, Monascus sp. has the advantages of safety and reliability, so that relevant immune preparations, hypoglyce- mic preparations and anti-aging preparations have great applica- tion potentials and huge economic values.
The present invention is intended to provide Monascuskaolin
A, B and C having glycosidase inhibitory, immunoregulatory and an- ti-aging activities, and a preparation method for Monascuskaolin
A, B and C.
Monascuskaolin A, B and C having glycosidase inhibitory, im- munoregulatory and anti-aging activities are separated and pre- pared from a Monascus sp. culture, and structural formulas of the
Monascuskaolin A, B and C are as follows:
A a, a ae en 5 Si J ete eds C. ee ë Ex a Bow 4 vi the Monascuskaolin A is red powder, with a molecular formula of C:5H::0:; the Monascuskaolin A has a glycosidase inhibitory ac- tivity, a median inhibition concentration (ICs) thereof against o- glycosidase is 220 pg/mL, and the Monascuskaolin A can signifi- cantly downregulate transcript levels of IL-1, TNF-« and TGF-p of in vitro cultured mouse macrophages RAW264.7: IL-1p (0.01+0.01),
TNF-a (0.0240.01) and TGF-B (0.03£0.01) which change significantly compared to a DMSO control group (P<0.05); a life span of fruit flies is extended by 31.6%; the Monascuskaolin B is red powder, with a molecular formula of C:3H:905; the Monascuskaolin B has a glycosidase inhibitory ac- tivity, a median inhibition concentration (IC) thereof against o- glycosidase is 174 pg/mL, and the Monascuskaolin B can signifi- cantly downregulate transcript levels of IL-1p, TNF-a and TGF-$ of in vitro cultured mouse macrophages RAW264.7: IL-1p (0.01+0.01),
TNF-o {0.02+£0.01) and TGF-B (0.03+0.01) which change significantly compared to a DMSO control group (P<0.05); a life span of fruit flies is extended by 27.3%; and the Monascuskaclin C is red powder, with a molecular formula of C3H.,05; the Monascuskaolin C has a glycosidase inhibitory ac- tivity, a median inhibition concentration (IC.y) thereof against o- glycosidase is 200 pg/mL, and the Monascuskaolin C can signifi- cantly downregulate transcript levels of IL-1, TNF-a and TGF-6 of in vitro cultured macrophages RAW264.7 of a mouse: IL-1p (0.01£0.01), TNF-o (0.02+0.01) and TGF-8 (0.03£0.01) which change significantly compared to a DMSO control group (P<0.05); a life span of fruit flies is extended by 22.5%.
The Monascus sp. 1s Monascus purpureus or Monascus anka or
Monascus ruber.
The Monascuskaolin A, B and C having glycosidase inhibitory, immunoregulatory and anti-aging activities are prepared by the following operating steps: (1) preparation of a solid culture performing primary strain culturing, secondary seed culturing and tertiary liquid amplification culturing on a strain of Monas- cus sp. to obtain a liquid culture; (1.1) primary strain culturing inoculating a spore suspension of Monascus sp. to a potato dextrose agar (PDA) slant culture medium, with an inoculation amount of 10-30 pL on each slant, and culturing the spore suspen- sion at a temperature of 32-35°C and a rotating speed of 220-250 r/min for 7-9 d to obtain a primary strain; (1.2) secondary seed culturing preparing the primary strain and sterile water into a spore suspension with a concentration of 10°-10" spores/mL, inoculating the spore suspension to a PDB culture medium, with an inoculation amount of 200-250 pL in each bottle, and performing shake-flask culturing at a temperature of 32-35°C and a rotating speed of 220- 250 r/min for 3-5 d to obtain a liquid secondary strain; (1.3) tertiary liquid amplification culturing inoculating 1-3 mL of liquid secondary strain to 250 mL of
PDB culture medium, performing shake-flask culturing at a tempera- ture of 32-35°C and a rotating speed of 220-250 r/min for 7-9 d, and collecting the culture to obtain a liquid culture; (2) extraction and refining of effective components in the liquid culture an extractant for extraction and refining is prepared by evenly mixing methanol and ethyl acetate according to a volume ra- tic of 0-1:1-0; specific operations are as follows: (2.1) extraction of effective components in the liquid cul- ture after decompression concentration and drying, performing ul- trasonic extraction on the liquid culture by using an extractant, filtrating by a filtration membrane or centrifuging the liquid culture, and removing the solvent by reduced pressure evaporation to obtain an effective component extract; (2.2) preliminary purification of the extract preliminarily purifying the effective component extract by using a preparative reversed-phase high performance liquid chro- matograph to obtain a reddish-brown preliminarily purified prod- uct; (2.3) refining of the preliminarily purified product refining the preliminarily purified product by using a semi- preparative reversed-phase high performance liquid chromatograph, collecting eluents with mass-to-charge ratios (m/z) corresponding to peaks 445, 403 and 331 under an acetonitrile solution eluting condition, respectively a Monascuskaolin A eluent, a Monascuskao- lin B eluent and a Monascuskaolin C eluent, to obtain red Monas- cuskaolin A, B and C.
Further preparation technical requirements are as follows:
In step (1.1), the PDA culture medium is a potato dextrose agar medium, wherein the formula of the PDA culture medium com- prises 200 g of peeled potato, 20 g of glucose and 15 g of agar, and water is added to 1,000 mL.
In steps (1.2) and (1.3), the formula of the PDB culture me- dium comprises 200 g of peeled potato and 20 g of glucose, and wa- ter is added to 1,000 mL.
In step (2.1), the liquid culture is concentrated by decom- pression to 3/10-1/10 of an original volume at a vacuum degree of -0.1 to -0.08 MP and a temperature of 10-60°C, dried at a tempera- ture of -50-130°C and crushed, the extractant is added into the 5 liquid culture according to a weight-to-volume ratio of 1 g:0.5-5 mL, and 40 KHz ultrasonic extraction is performed for 20-200 min; the extract is centrifuged at 4,000-15,000 r/min or filtrated by a 0.1-2.5 pm filtration membrane to obtain a filtrate or a centri- fuged supernatant; and the extractant is removed from the filtrate or the centrifuged supernatant through reduced pressure evapora- tion at a vacuum degree of -0.1 to -0.08 MP and a temperature of 10-60°C to obtain the effective component extract.
In step (2.2), the preparative reversed-phase high perfor- mance liquid chromatograph is used for refining; the chromato- graphic separation preparation conditions are as follows: a mobile phase A is ultrapure water, and a mobile phase B is acetonitrile; eluting conditions: gradient elution is performed with the 50%-70% mobile phase B from 0 min to 30 min; an injection volume is 30-300 pL; a column temperature is 20-40°C; a flow rate is 5-50 mL/min; a chromatographic column is a preparative reversed-phase C-18 col- umn; the eluents with mass-to-charge ratios (m/z) corresponding to peaks 445, 403 and 331 are respectively collected; the extractant is removed through reduced pressure evaporation at a vacuum degree of -0.1 to -0.08 MP and a temperature of 10-60°C; the precipitates are dried at a temperature of lower than or equal to 130°C to ob- tain the red preliminarily purified crude products A, B and C, re- spectively.
In step (2.3), the chromatographic separation preparation conditions are as follows: a mobile phase A is ultrapure water, and a mobile phase B is acetonitrile; eluting conditions: elution is performed with the 50%-70% mobile phase A from 0 min to 30 min; an injection volume is 20-200 pL; a column temperature is 20-40°C; a flow rate is 2-20 mL/min; a chromatographic column is a semipre- parative reversed-phase C-18 column; the crude product A, the crude product B and the crude product C are respectively injected and the eluents with mass-to-charge ratios (m/z) corresponding to peaks 445, 403 and 331 are respectively collected; the extractant is removed through reduced pressure evapcration at a vacuum degree of -0.1 to -0.08 MP and a temperature of 10-60°C; the precipitates are dried at a temperature of lower than or equal to 130°C to ob- tain the red solid Monascuskaolin A, B and C.
In steps (2.1-2.3), the extractant is methanol or ethyl ace- tate or a mixture of methanol and ethyl acetate.
Analytical research of the present invention is described as follows: 1. The extract is separated and purified by means of a pre- parative reversed-phase high performance liquid chromatography and a semipreparative reversed-phase high performance liquid chroma- tography to obtain 3 novel compounds from Monascus sp., named
Monascuskaolin A, B and C. 2. The Monascuskaolin A has a glycosidase inhibitory activi- ty, a median inhibition concentration (IC) thereof against o- glycosidase is 220 u9/mL, and the Monascuskaolin A can signifi- cantly downregulate transcript levels of IL-1f, TNF-a and TGF-p of in vitro cultured mouse macrophages RAW264.7: IL-1B8 (0.01+0.01),
TNF-0 (0.02+0.01) and TGF-B {0.03+0.01) which change significantly compared to a DMSO control group (P<0.00); a life span of fruit flies is extended by 31.63; the Monascuskaolin B is red powder, with a molecular formula of C:3H306 and a glycosidase inhibitory activity, a median inhibi- tion concentration {ICs;) thereof against o-glycosidase is 174 ng/mL, and the Monascuskaolin B can significantly downregulate transcript levels of IL-18, TNF-« and TGF-p of in vitro cultured mouse macrophages RAW264.7: IL-1B8 (0.01+0.01), TNF-o (0.02x0.01) and TGF-B (0.03+£0.01) which change significantly compared to a
DMSO control group (P<0.05); a life span of fruit flies is extend- ed by 27.3%; and the Monascuskaolin C has a glycosidase inhibitory activity, a median inhibition concentration (ICs) against o-glycosidase there- of is 200 pg/mL, and the Monascuskaolin C can significantly down- regulate transcript levels of IL-18, TNF-a and TGF-B of in vitro cultured mouse macrophages RAW264.7: IL-1p (0.0140.01), TNF-Q (0.02+0.01) and TGF-p (0.03+0.01) which change significantly com-
pared to a DMSO control group (P<0.05); a life span of fruit flies is extended by 22.5%. 4. Chemical structure identification data of Monascuskaolin
A, B and C
It is shown by high resolution liquid chromatography-mass spectrometry that a mass-to-charge ratio of cations [M + H] of the Monascuskaolin A is 445.2224, and a corresponding molecular formula is C::H::0;. The specific data of NMR spectrometry are shown in Table 1.
Table 1. Attribution of ‘H-NMR and ‘C-NMR spectrometry of
Monascuskaolin A
No. | Carbon DEPT Hydrogen shift COSY HMBC NOESY shift HMQC | value 8, mult (J value 6 in Hz) 1 148.4 CH 7.51, s 42.4, 115.0, 146.4, 3.94 em pe
Swe jo OO 4 108.7 CH 6.32,s 39.2, 104.9, 115.0, 1.28,2.71,5.38
A + wwe oT es fm cms je ze sm
EC ee ae 42.7 CH 3.94, br 4.17 22.1, 55.8, 82.8, 1.52, 7.51 115.0, 146.4, 148.4, 192.8, 202.7 11 39.2 CH, 2.71, d 5.15 18.7, 68.1, 108.7, 1.28, 5.15, 6.32
Co a ee fe
7.2) 2.57,1.57 2.98 dt (18.2, 7.1) 16 22.7 CH, 1.57, quint {7.1} | 2.57,2.98, | 202.7,42.4,28.7 1.26, 2.57, 2.98 17 [287 Ch, 1.26, m 1.57
EE zz EZ sa aje as Jw ae | jw 21 | 13.1 CH, 0.89, t{7.1) 1.28 31.5, 22,3
En
It is shown by high resolution liquid chromatography-mass spectrometry that a mass-to-charge ratio of cations [M + H] © of the Monascuskaclin B is 403.2199, and a corresponding molecular formula is C:3H:0s. The specific data of NMR spectrometry are shown in Table 2.
Table 2. Attribution of ‘H-NMR and “C-NMR spectrometry of
Monascuskaolin B } oe eee -
DEPT shift value value ô,, mult COSY HMBC
HMQC êc {/ in Hz) eee ea [a wwe fe | 1
Ses jo [we | jmewme es eT ce ser Jo 9a 42.7 CH 3.93,s 4.04 22.1, 55.8, 82.8, 115.0, 147.0, en 5 mr fe |] 15 42.4 CH; 2.51, dt (18.2, 2.98, 1.51 202.7,22.7,28.7 7.2) 2.57, 1.51 2.98 dt (18.2, 7.1) 16 22.7 CH, 1.57, quint {7.1} | 2.51, 2.98, | 202.7,42.4,28.7
KG ea aan ze
It is shown by high resolution liquid chromatography-mass spectrometry that a mass-to-charge ratio of cations [M + H] 7 of the Monascuskaolin C is 331.1565, and a corresponding molecular formula is C:5H::0s. The specific data of NMR spectrometry are shown in Table 3.
Table 3. Attribution of ‘H-NMR and “C-NMR spectrometry of
Monascuskaolin C
Hydrogen shift shift DEPT value §,, mult (Jin | COSY HMBC value HMQC
Hz) bc ete aan fr
I te wwe er
KE mw
TT ee alata [amen a ee 13 1422 CH, 2.52, dt {18.2, 7.2} | 2.97,1.55 202.7, 31.0
CU naan aa
The present invention has the following beneficial technical effects: 1. To look for novel, natural and efficient substances having glycosidase inhibitory, immunoregulatory and anti-aging activi- ties, the inventors cultured a plenty of Monascus sp. strains at first and then performed extraction and activity determination on the effective components of the culture, and discovered that a
Monascus sp. strain culture extract had the glycosidase inhibito- ry, RAW264.7 cellular inflammation inhibitory, immune activities and had an anti-aging activity. Based on massive culturing and ex- traction, Monascuskaolin A, B and C were obtained through separa- tion and purification by means of preparative reversed-phase chro- matography and semipreparative reversed-phase chromatography.
These compounds are reddish-brown powder and structural formulas of Monascuskaolin A, B and C were obtained by 1D-NMR spectrometry, 2D-NMR spectrometry and accurate mass spectrometry. They have strong o-glycosidase inhibitory, RAW264.7 cellular immunosuppres- sive and anti-aging activities and have the potential for develop- ing diabetes drugs, anti-inflammatory and immune-related drugs and anti-aging preparations. 2. In the present invention, a Monascus sp. strain was first discovered to have the function of metabolizing the above- mentioned active substances. On the basis of the present inven- tion, related genes are expected to be further cloned to construct high-yield strains. 3. As a food microorganism, Monascus sp. has the advantages of safety and reliability, so that relevant immune preparations,
hypoglycemic preparations and anti-aging preparations have great application potentials and huge economic values. 4. As the Monascuskaolin A, B and C are produced by means of microbial fermentation, the preparation method is not affected by natural environment and resources, and is liable to realize indus- trial, automatic and continuous production without causing damages to natural environment and natural resources; the Monascuskaolin products produced by the process method of the present invention are low in cost, and the preparation process is simple, stable, easy to regulate and high in success rate.
Claims (1)
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211287602.0A CN115611914A (en) | 2022-10-20 | 2022-10-20 | Monascus A, B and C with glycosidase inhibiting activity, immunoregulation activity and anti-aging activity and preparation method thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| NL2033900B1 true NL2033900B1 (en) | 2024-05-08 |
Family
ID=84865517
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| NL2033900A NL2033900B1 (en) | 2022-10-20 | 2023-01-02 | Monascuskaolin a, b and c having glycosidase inhibitory, immunoregulatory and anti-aging activities and preparation method thereof |
Country Status (2)
| Country | Link |
|---|---|
| CN (1) | CN115611914A (en) |
| NL (1) | NL2033900B1 (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114478565A (en) * | 2021-12-08 | 2022-05-13 | 安徽农业大学 | Monascus A with glycosidase inhibiting activity and immunoregulation activity and preparation method thereof |
| CN114478564A (en) * | 2021-12-08 | 2022-05-13 | 安徽农业大学 | Monascus C with glycosidase inhibiting activity and immunoregulation activity and preparation method thereof |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114292280A (en) * | 2021-12-08 | 2022-04-08 | 安徽农业大学 | Monascus B with glycosidase inhibiting activity and immunoregulation activity and preparation method thereof |
-
2022
- 2022-10-20 CN CN202211287602.0A patent/CN115611914A/en active Pending
-
2023
- 2023-01-02 NL NL2033900A patent/NL2033900B1/en active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114478565A (en) * | 2021-12-08 | 2022-05-13 | 安徽农业大学 | Monascus A with glycosidase inhibiting activity and immunoregulation activity and preparation method thereof |
| CN114478564A (en) * | 2021-12-08 | 2022-05-13 | 安徽农业大学 | Monascus C with glycosidase inhibiting activity and immunoregulation activity and preparation method thereof |
Non-Patent Citations (2)
| Title |
|---|
| BALAKRISHNAN BIJINU ET AL: "Mpp7 controls regioselective Knoevenagel condensation during the biosynthesis ofMonascusazaphilone pigments", TETRAHEDRON LETTERS, vol. 55, no. 9, 2014, pages 1640 - 1643, XP028615088, ISSN: 0040-4039, DOI: 10.1016/J.TETLET.2014.01.090 * |
| PROMSUK GUNLATIDA ET AL: "Absolute configuration of azaphilones from Monascus kaoliangKB9 and solvent effects on their keto and enol forms.", NATURAL PRODUCT RESEARCH 10 FEB 2022, 10 February 2022 (2022-02-10), pages 1 - 8, XP009544713, ISSN: 1478-6427 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN115611914A (en) | 2023-01-17 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN110283226B (en) | Method for extracting antioxidant component in rosemary | |
| CN114478565B (en) | Monascin A with glycosidase activity inhibiting and immunoregulatory activity and preparation method thereof | |
| CN108558817B (en) | Isocoumarin derivative and preparation method and application thereof | |
| CN109400664B (en) | Method for extracting rice oil phytosterol | |
| CN115073295A (en) | Unsaturated fatty acid in purslane, and extraction and separation method and application thereof | |
| KR20210058416A (en) | Novel Aspergillus tubingensis C2-2 isolated from Nu-ruk producing ginsenoside compound K biotransformation enzyme and use thereof | |
| NL2033900B1 (en) | Monascuskaolin a, b and c having glycosidase inhibitory, immunoregulatory and anti-aging activities and preparation method thereof | |
| Moreau et al. | Production of eremofortins A, B, and C relative to formation of PR toxin by Penicillium roqueforti | |
| CN114478564B (en) | Monascin C with glycosidase activity inhibiting and immunoregulatory activity and preparation method thereof | |
| CN108186790B (en) | Rehmannia root extract, preparation method and application in promoting CIK cell in-vitro proliferation | |
| CN108384814B (en) | Preparation method of phloretin | |
| CN114292280A (en) | Monascus B with glycosidase inhibiting activity and immunoregulation activity and preparation method thereof | |
| CN113754526A (en) | High-purity coenzyme Q10 purification process | |
| CN101117641B (en) | Method for preparing secoisolariciresinol diglucoside | |
| KR101767337B1 (en) | Novel pure substance extracting method | |
| CN109022512B (en) | Method for biologically synthesizing Hispidin | |
| CN105420293A (en) | Method for separating and purifying resveratrol from traditional Chinese medicine polygonum cuspidatum extraction solution | |
| CN101928291B (en) | Method for separating and purifying ansamitocin from precious orange synnema actinomycetes fermentation liquor | |
| KR20210017898A (en) | Mass production method of luteolin | |
| JP2004033088A (en) | Method for producing jasmonic acids and theobroxide by fungus | |
| CN115537434B (en) | Method for preparing tectorigenin from Sichuan blackberry lily | |
| CN108586435A (en) | The preparation method of Echinulin | |
| CN115490588B (en) | Method for separating various unsaturated fatty acids from torreya seed oil | |
| CN116063262B (en) | Kojic acid compound in noni fruits and preparation method and application thereof | |
| CN114456138B (en) | Method for separating three coumarin compounds from fingered citron extract |