CN116410201A - Flavonoid compound extracted from black mulberry plants and preparation method and application thereof - Google Patents
Flavonoid compound extracted from black mulberry plants and preparation method and application thereof Download PDFInfo
- Publication number
- CN116410201A CN116410201A CN202111644417.8A CN202111644417A CN116410201A CN 116410201 A CN116410201 A CN 116410201A CN 202111644417 A CN202111644417 A CN 202111644417A CN 116410201 A CN116410201 A CN 116410201A
- Authority
- CN
- China
- Prior art keywords
- water
- ethanol
- preparation
- column
- component
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 229930003935 flavonoid Natural products 0.000 title claims abstract description 46
- 235000017173 flavonoids Nutrition 0.000 title claims abstract description 46
- 238000002360 preparation method Methods 0.000 title claims abstract description 38
- -1 Flavonoid compound Chemical class 0.000 title claims abstract description 28
- 240000000249 Morus alba Species 0.000 title claims abstract description 12
- 238000004128 high performance liquid chromatography Methods 0.000 claims abstract description 25
- 150000003839 salts Chemical class 0.000 claims abstract description 20
- 238000004440 column chromatography Methods 0.000 claims abstract description 11
- 238000000605 extraction Methods 0.000 claims abstract description 10
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 88
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 57
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 36
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 24
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 24
- 150000002215 flavonoids Chemical class 0.000 claims description 23
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 22
- YASAKCUCGLMORW-UHFFFAOYSA-N Rosiglitazone Chemical compound C=1C=CC=NC=1N(C)CCOC(C=C1)=CC=C1CC1SC(=O)NC1=O YASAKCUCGLMORW-UHFFFAOYSA-N 0.000 claims description 22
- 239000000243 solution Substances 0.000 claims description 15
- 208000001072 type 2 diabetes mellitus Diseases 0.000 claims description 14
- 239000008194 pharmaceutical composition Substances 0.000 claims description 13
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 12
- 239000003480 eluent Substances 0.000 claims description 12
- 239000000499 gel Substances 0.000 claims description 12
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 11
- 206010022489 Insulin Resistance Diseases 0.000 claims description 11
- 229960004586 rosiglitazone Drugs 0.000 claims description 11
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 10
- 238000000034 method Methods 0.000 claims description 10
- 239000000741 silica gel Substances 0.000 claims description 10
- 229910002027 silica gel Inorganic materials 0.000 claims description 10
- 238000010828 elution Methods 0.000 claims description 9
- 238000010898 silica gel chromatography Methods 0.000 claims description 9
- 239000011259 mixed solution Substances 0.000 claims description 8
- 239000003208 petroleum Substances 0.000 claims description 8
- 239000000843 powder Substances 0.000 claims description 8
- 230000002441 reversible effect Effects 0.000 claims description 8
- 235000008708 Morus alba Nutrition 0.000 claims description 7
- 239000012046 mixed solvent Substances 0.000 claims description 7
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims description 6
- 238000001514 detection method Methods 0.000 claims description 6
- 239000003814 drug Substances 0.000 claims description 6
- 208000030159 metabolic disease Diseases 0.000 claims description 6
- 238000002156 mixing Methods 0.000 claims description 6
- 239000000203 mixture Substances 0.000 claims description 6
- 230000008569 process Effects 0.000 claims description 6
- 201000010099 disease Diseases 0.000 claims description 5
- 239000000546 pharmaceutical excipient Substances 0.000 claims description 5
- 239000011347 resin Substances 0.000 claims description 5
- 229920005989 resin Polymers 0.000 claims description 5
- ZOBPZXTWZATXDG-UHFFFAOYSA-N 1,3-thiazolidine-2,4-dione Chemical compound O=C1CSC(=O)N1 ZOBPZXTWZATXDG-UHFFFAOYSA-N 0.000 claims description 4
- 229940123464 Thiazolidinedione Drugs 0.000 claims description 4
- 239000012043 crude product Substances 0.000 claims description 4
- 239000000945 filler Substances 0.000 claims description 4
- 229940124531 pharmaceutical excipient Drugs 0.000 claims description 4
- 230000005526 G1 to G0 transition Effects 0.000 claims description 3
- 159000000007 calcium salts Chemical class 0.000 claims description 3
- 238000001035 drying Methods 0.000 claims description 3
- 238000001914 filtration Methods 0.000 claims description 3
- 239000007788 liquid Substances 0.000 claims description 3
- 208000024172 Cardiovascular disease Diseases 0.000 claims description 2
- 108010010803 Gelatin Proteins 0.000 claims description 2
- 208000031226 Hyperlipidaemia Diseases 0.000 claims description 2
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 claims description 2
- 229920000168 Microcrystalline cellulose Polymers 0.000 claims description 2
- 229920000881 Modified starch Polymers 0.000 claims description 2
- GHUUBYQTCDQWRA-UHFFFAOYSA-N Pioglitazone hydrochloride Chemical compound Cl.N1=CC(CC)=CC=C1CCOC(C=C1)=CC=C1CC1C(=O)NC(=O)S1 GHUUBYQTCDQWRA-UHFFFAOYSA-N 0.000 claims description 2
- 229930006000 Sucrose Natural products 0.000 claims description 2
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 2
- 239000004480 active ingredient Substances 0.000 claims description 2
- 239000002775 capsule Substances 0.000 claims description 2
- 239000008273 gelatin Substances 0.000 claims description 2
- 229920000159 gelatin Polymers 0.000 claims description 2
- 235000019322 gelatine Nutrition 0.000 claims description 2
- 235000011852 gelatine desserts Nutrition 0.000 claims description 2
- 239000008187 granular material Substances 0.000 claims description 2
- 238000002347 injection Methods 0.000 claims description 2
- 239000007924 injection Substances 0.000 claims description 2
- 239000008101 lactose Substances 0.000 claims description 2
- 235000019813 microcrystalline cellulose Nutrition 0.000 claims description 2
- 239000008108 microcrystalline cellulose Substances 0.000 claims description 2
- 229940016286 microcrystalline cellulose Drugs 0.000 claims description 2
- 229960002827 pioglitazone hydrochloride Drugs 0.000 claims description 2
- 239000005720 sucrose Substances 0.000 claims description 2
- 238000009472 formulation Methods 0.000 claims 1
- 230000002265 prevention Effects 0.000 claims 1
- 239000003826 tablet Substances 0.000 claims 1
- 230000000694 effects Effects 0.000 abstract description 19
- 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 abstract description 9
- 239000008103 glucose Substances 0.000 abstract description 9
- 108010076365 Adiponectin Proteins 0.000 abstract description 6
- 102000011690 Adiponectin Human genes 0.000 abstract description 6
- 239000012530 fluid Substances 0.000 abstract description 4
- NOESYZHRGYRDHS-UHFFFAOYSA-N insulin Chemical compound N1C(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(NC(=O)CN)C(C)CC)CSSCC(C(NC(CO)C(=O)NC(CC(C)C)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CCC(N)=O)C(=O)NC(CC(C)C)C(=O)NC(CCC(O)=O)C(=O)NC(CC(N)=O)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CSSCC(NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2C=CC(O)=CC=2)NC(=O)C(CC(C)C)NC(=O)C(C)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2NC=NC=2)NC(=O)C(CO)NC(=O)CNC2=O)C(=O)NCC(=O)NC(CCC(O)=O)C(=O)NC(CCCNC(N)=N)C(=O)NCC(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC(O)=CC=3)C(=O)NC(C(C)O)C(=O)N3C(CCC3)C(=O)NC(CCCCN)C(=O)NC(C)C(O)=O)C(=O)NC(CC(N)=O)C(O)=O)=O)NC(=O)C(C(C)CC)NC(=O)C(CO)NC(=O)C(C(C)O)NC(=O)C1CSSCC2NC(=O)C(CC(C)C)NC(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(N)CC=1C=CC=CC=1)C(C)C)CC1=CN=CN1 NOESYZHRGYRDHS-UHFFFAOYSA-N 0.000 description 20
- 150000001875 compounds Chemical class 0.000 description 15
- 102000004877 Insulin Human genes 0.000 description 10
- 108090001061 Insulin Proteins 0.000 description 10
- 230000004069 differentiation Effects 0.000 description 10
- 229940125396 insulin Drugs 0.000 description 10
- 239000012071 phase Substances 0.000 description 8
- 239000002253 acid Substances 0.000 description 7
- 210000004027 cell Anatomy 0.000 description 7
- 208000035475 disorder Diseases 0.000 description 7
- 229930182733 sanggenol Natural products 0.000 description 7
- 210000001789 adipocyte Anatomy 0.000 description 6
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 6
- 210000000229 preadipocyte Anatomy 0.000 description 6
- HIXDQWDOVZUNNA-UHFFFAOYSA-N 2-(3,4-dimethoxyphenyl)-5-hydroxy-7-methoxychromen-4-one Chemical compound C=1C(OC)=CC(O)=C(C(C=2)=O)C=1OC=2C1=CC=C(OC)C(OC)=C1 HIXDQWDOVZUNNA-UHFFFAOYSA-N 0.000 description 5
- 238000002835 absorbance Methods 0.000 description 5
- 238000006243 chemical reaction Methods 0.000 description 5
- 230000006698 induction Effects 0.000 description 5
- 238000007789 sealing Methods 0.000 description 5
- 238000004809 thin layer chromatography Methods 0.000 description 5
- CSCPPACGZOOCGX-MICDWDOJSA-N 1-deuteriopropan-2-one Chemical compound [2H]CC(C)=O CSCPPACGZOOCGX-MICDWDOJSA-N 0.000 description 4
- VZCYOOQTPOCHFL-OWOJBTEDSA-N Fumaric acid Chemical compound OC(=O)\C=C\C(O)=O VZCYOOQTPOCHFL-OWOJBTEDSA-N 0.000 description 4
- 238000003556 assay Methods 0.000 description 4
- 239000008280 blood Substances 0.000 description 4
- 210000004369 blood Anatomy 0.000 description 4
- 238000004113 cell culture Methods 0.000 description 4
- 239000003153 chemical reaction reagent Substances 0.000 description 4
- 239000003795 chemical substances by application Substances 0.000 description 4
- 239000000463 material Substances 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 3
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 3
- 108090000790 Enzymes Proteins 0.000 description 3
- MUBZPKHOEPUJKR-UHFFFAOYSA-N Oxalic acid Chemical compound OC(=O)C(O)=O MUBZPKHOEPUJKR-UHFFFAOYSA-N 0.000 description 3
- OFOBLEOULBTSOW-UHFFFAOYSA-N Propanedioic acid Natural products OC(=O)CC(O)=O OFOBLEOULBTSOW-UHFFFAOYSA-N 0.000 description 3
- 230000002378 acidificating effect Effects 0.000 description 3
- 229930013930 alkaloid Natural products 0.000 description 3
- 238000010009 beating Methods 0.000 description 3
- 230000008859 change Effects 0.000 description 3
- UREBDLICKHMUKA-CXSFZGCWSA-N dexamethasone Chemical compound C1CC2=CC(=O)C=C[C@]2(C)[C@]2(F)[C@@H]1[C@@H]1C[C@@H](C)[C@@](C(=O)CO)(O)[C@@]1(C)C[C@@H]2O UREBDLICKHMUKA-CXSFZGCWSA-N 0.000 description 3
- 229960003957 dexamethasone Drugs 0.000 description 3
- 229940079593 drug Drugs 0.000 description 3
- 229940088598 enzyme Drugs 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 239000012091 fetal bovine serum Substances 0.000 description 3
- 125000000524 functional group Chemical group 0.000 description 3
- 150000002632 lipids Chemical class 0.000 description 3
- 239000002609 medium Substances 0.000 description 3
- 239000012528 membrane Substances 0.000 description 3
- FJKROLUGYXJWQN-UHFFFAOYSA-N papa-hydroxy-benzoic acid Natural products OC(=O)C1=CC=C(O)C=C1 FJKROLUGYXJWQN-UHFFFAOYSA-N 0.000 description 3
- 239000002904 solvent Substances 0.000 description 3
- VZCYOOQTPOCHFL-UHFFFAOYSA-N trans-butenedioic acid Natural products OC(=O)C=CC(O)=O VZCYOOQTPOCHFL-UHFFFAOYSA-N 0.000 description 3
- GHOKWGTUZJEAQD-ZETCQYMHSA-N (D)-(+)-Pantothenic acid Chemical compound OCC(C)(C)[C@@H](O)C(=O)NCCC(O)=O GHOKWGTUZJEAQD-ZETCQYMHSA-N 0.000 description 2
- WXTMDXOMEHJXQO-UHFFFAOYSA-N 2,5-dihydroxybenzoic acid Chemical compound OC(=O)C1=CC(O)=CC=C1O WXTMDXOMEHJXQO-UHFFFAOYSA-N 0.000 description 2
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 2
- RGHNJXZEOKUKBD-SQOUGZDYSA-N D-gluconic acid Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C(O)=O RGHNJXZEOKUKBD-SQOUGZDYSA-N 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- 108010001336 Horseradish Peroxidase Proteins 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 2
- AFVFQIVMOAPDHO-UHFFFAOYSA-N Methanesulfonic acid Chemical compound CS(O)(=O)=O AFVFQIVMOAPDHO-UHFFFAOYSA-N 0.000 description 2
- 241000218231 Moraceae Species 0.000 description 2
- 241000218213 Morus <angiosperm> Species 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- 206010070834 Sensitisation Diseases 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- 150000007513 acids Chemical class 0.000 description 2
- 150000003797 alkaloid derivatives Chemical class 0.000 description 2
- 125000003118 aryl group Chemical group 0.000 description 2
- WPYMKLBDIGXBTP-UHFFFAOYSA-N benzoic acid Chemical compound OC(=O)C1=CC=CC=C1 WPYMKLBDIGXBTP-UHFFFAOYSA-N 0.000 description 2
- 238000004587 chromatography analysis Methods 0.000 description 2
- 229960004106 citric acid Drugs 0.000 description 2
- 239000013256 coordination polymer Substances 0.000 description 2
- ZYGHJZDHTFUPRJ-UHFFFAOYSA-N coumarin Chemical compound C1=CC=C2OC(=O)C=CC2=C1 ZYGHJZDHTFUPRJ-UHFFFAOYSA-N 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- XBDQKXXYIPTUBI-UHFFFAOYSA-N dimethylselenoniopropionate Natural products CCC(O)=O XBDQKXXYIPTUBI-UHFFFAOYSA-N 0.000 description 2
- 239000001530 fumaric acid Substances 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 239000012442 inert solvent Substances 0.000 description 2
- KQNPFQTWMSNSAP-UHFFFAOYSA-N isobutyric acid Chemical compound CC(C)C(O)=O KQNPFQTWMSNSAP-UHFFFAOYSA-N 0.000 description 2
- TWBYWOBDOCUKOW-UHFFFAOYSA-N isonicotinic acid Chemical compound OC(=O)C1=CC=NC=C1 TWBYWOBDOCUKOW-UHFFFAOYSA-N 0.000 description 2
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 2
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 2
- 125000001434 methanylylidene group Chemical group [H]C#[*] 0.000 description 2
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 2
- 125000001570 methylene group Chemical group [H]C([H])([*:1])[*:2] 0.000 description 2
- XNGIFLGASWRNHJ-UHFFFAOYSA-N phthalic acid Chemical compound OC(=O)C1=CC=CC=C1C(O)=O XNGIFLGASWRNHJ-UHFFFAOYSA-N 0.000 description 2
- 239000011148 porous material Substances 0.000 description 2
- 230000001737 promoting effect Effects 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- YGSDEFSMJLZEOE-UHFFFAOYSA-N salicylic acid Chemical compound OC(=O)C1=CC=CC=C1O YGSDEFSMJLZEOE-UHFFFAOYSA-N 0.000 description 2
- 230000008313 sensitization Effects 0.000 description 2
- 238000001228 spectrum Methods 0.000 description 2
- 239000012089 stop solution Substances 0.000 description 2
- TYFQFVWCELRYAO-UHFFFAOYSA-N suberic acid Chemical compound OC(=O)CCCCCCC(O)=O TYFQFVWCELRYAO-UHFFFAOYSA-N 0.000 description 2
- 208000024891 symptom Diseases 0.000 description 2
- JOXIMZWYDAKGHI-UHFFFAOYSA-N toluene-4-sulfonic acid Chemical compound CC1=CC=C(S(O)(=O)=O)C=C1 JOXIMZWYDAKGHI-UHFFFAOYSA-N 0.000 description 2
- QBYIENPQHBMVBV-HFEGYEGKSA-N (2R)-2-hydroxy-2-phenylacetic acid Chemical compound O[C@@H](C(O)=O)c1ccccc1.O[C@@H](C(O)=O)c1ccccc1 QBYIENPQHBMVBV-HFEGYEGKSA-N 0.000 description 1
- WRIDQFICGBMAFQ-UHFFFAOYSA-N (E)-8-Octadecenoic acid Natural products CCCCCCCCCC=CCCCCCCC(O)=O WRIDQFICGBMAFQ-UHFFFAOYSA-N 0.000 description 1
- TUSDEZXZIZRFGC-UHFFFAOYSA-N 1-O-galloyl-3,6-(R)-HHDP-beta-D-glucose Natural products OC1C(O2)COC(=O)C3=CC(O)=C(O)C(O)=C3C3=C(O)C(O)=C(O)C=C3C(=O)OC1C(O)C2OC(=O)C1=CC(O)=C(O)C(O)=C1 TUSDEZXZIZRFGC-UHFFFAOYSA-N 0.000 description 1
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 1
- 238000005160 1H NMR spectroscopy Methods 0.000 description 1
- MZPDTOMKQCMETI-UHFFFAOYSA-N 2,6-dimethylocta-2,6-diene Chemical compound CC=C(C)CCC=C(C)C MZPDTOMKQCMETI-UHFFFAOYSA-N 0.000 description 1
- LQJBNNIYVWPHFW-UHFFFAOYSA-N 20:1omega9c fatty acid Natural products CCCCCCCCCCC=CCCCCCCCC(O)=O LQJBNNIYVWPHFW-UHFFFAOYSA-N 0.000 description 1
- ALKYHXVLJMQRLQ-UHFFFAOYSA-N 3-Hydroxy-2-naphthoate Chemical compound C1=CC=C2C=C(O)C(C(=O)O)=CC2=C1 ALKYHXVLJMQRLQ-UHFFFAOYSA-N 0.000 description 1
- HVCOBJNICQPDBP-UHFFFAOYSA-N 3-[3-[3,5-dihydroxy-6-methyl-4-(3,4,5-trihydroxy-6-methyloxan-2-yl)oxyoxan-2-yl]oxydecanoyloxy]decanoic acid;hydrate Chemical compound O.OC1C(OC(CC(=O)OC(CCCCCCC)CC(O)=O)CCCCCCC)OC(C)C(O)C1OC1C(O)C(O)C(O)C(C)O1 HVCOBJNICQPDBP-UHFFFAOYSA-N 0.000 description 1
- BMYNFMYTOJXKLE-UHFFFAOYSA-N 3-azaniumyl-2-hydroxypropanoate Chemical compound NCC(O)C(O)=O BMYNFMYTOJXKLE-UHFFFAOYSA-N 0.000 description 1
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 description 1
- 229940090248 4-hydroxybenzoic acid Drugs 0.000 description 1
- QSBYPNXLFMSGKH-UHFFFAOYSA-N 9-Heptadecensaeure Natural products CCCCCCCC=CCCCCCCCC(O)=O QSBYPNXLFMSGKH-UHFFFAOYSA-N 0.000 description 1
- 239000004475 Arginine Substances 0.000 description 1
- 239000005711 Benzoic acid Substances 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical group [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- GHOKWGTUZJEAQD-UHFFFAOYSA-N Chick antidermatitis factor Natural products OCC(C)(C)C(O)C(=O)NCCC(O)=O GHOKWGTUZJEAQD-UHFFFAOYSA-N 0.000 description 1
- 235000004035 Cryptotaenia japonica Nutrition 0.000 description 1
- RGHNJXZEOKUKBD-UHFFFAOYSA-N D-gluconic acid Natural products OCC(O)C(O)C(O)C(O)C(O)=O RGHNJXZEOKUKBD-UHFFFAOYSA-N 0.000 description 1
- FEWJPZIEWOKRBE-JCYAYHJZSA-N Dextrotartaric acid Chemical compound OC(=O)[C@H](O)[C@@H](O)C(O)=O FEWJPZIEWOKRBE-JCYAYHJZSA-N 0.000 description 1
- 238000008157 ELISA kit Methods 0.000 description 1
- 239000001263 FEMA 3042 Substances 0.000 description 1
- 239000004366 Glucose oxidase Substances 0.000 description 1
- 108010015776 Glucose oxidase Proteins 0.000 description 1
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 description 1
- 229930186217 Glycolipid Natural products 0.000 description 1
- 206010060378 Hyperinsulinaemia Diseases 0.000 description 1
- ODKSFYDXXFIFQN-BYPYZUCNSA-P L-argininium(2+) Chemical compound NC(=[NH2+])NCCC[C@H]([NH3+])C(O)=O ODKSFYDXXFIFQN-BYPYZUCNSA-P 0.000 description 1
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 description 1
- FEWJPZIEWOKRBE-JCYAYHJZSA-M L-tartrate(1-) Chemical compound OC(=O)[C@H](O)[C@@H](O)C([O-])=O FEWJPZIEWOKRBE-JCYAYHJZSA-M 0.000 description 1
- 101000775476 Mus musculus Adiponectin Proteins 0.000 description 1
- GRYLNZFGIOXLOG-UHFFFAOYSA-N Nitric acid Chemical compound O[N+]([O-])=O GRYLNZFGIOXLOG-UHFFFAOYSA-N 0.000 description 1
- 239000005642 Oleic acid Substances 0.000 description 1
- ZQPPMHVWECSIRJ-UHFFFAOYSA-N Oleic acid Natural products CCCCCCCCC=CCCCCCCCC(O)=O ZQPPMHVWECSIRJ-UHFFFAOYSA-N 0.000 description 1
- LRBQNJMCXXYXIU-PPKXGCFTSA-N Penta-digallate-beta-D-glucose Natural products OC1=C(O)C(O)=CC(C(=O)OC=2C(=C(O)C=C(C=2)C(=O)OC[C@@H]2[C@H]([C@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)[C@@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)[C@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)O2)OC(=O)C=2C=C(OC(=O)C=3C=C(O)C(O)=C(O)C=3)C(O)=C(O)C=2)O)=C1 LRBQNJMCXXYXIU-PPKXGCFTSA-N 0.000 description 1
- 102000003992 Peroxidases Human genes 0.000 description 1
- IWYDHOAUDWTVEP-UHFFFAOYSA-N R-2-phenyl-2-hydroxyacetic acid Natural products OC(=O)C(O)C1=CC=CC=C1 IWYDHOAUDWTVEP-UHFFFAOYSA-N 0.000 description 1
- KDYFGRWQOYBRFD-UHFFFAOYSA-N Succinic acid Natural products OC(=O)CCC(O)=O KDYFGRWQOYBRFD-UHFFFAOYSA-N 0.000 description 1
- FEWJPZIEWOKRBE-UHFFFAOYSA-N Tartaric acid Natural products [H+].[H+].[O-]C(=O)C(O)C(O)C([O-])=O FEWJPZIEWOKRBE-UHFFFAOYSA-N 0.000 description 1
- GAMYVSCDDLXAQW-AOIWZFSPSA-N Thermopsosid Natural products O(C)c1c(O)ccc(C=2Oc3c(c(O)cc(O[C@H]4[C@H](O)[C@@H](O)[C@H](O)[C@H](CO)O4)c3)C(=O)C=2)c1 GAMYVSCDDLXAQW-AOIWZFSPSA-N 0.000 description 1
- 102000007641 Trefoil Factors Human genes 0.000 description 1
- 235000015724 Trifolium pratense Nutrition 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 235000011054 acetic acid Nutrition 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 210000000577 adipose tissue Anatomy 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- GZCGUPFRVQAUEE-VANKVMQKSA-N aldehydo-L-glucose Chemical compound OC[C@H](O)[C@H](O)[C@@H](O)[C@H](O)C=O GZCGUPFRVQAUEE-VANKVMQKSA-N 0.000 description 1
- AZDRQVAHHNSJOQ-UHFFFAOYSA-N alumane Chemical class [AlH3] AZDRQVAHHNSJOQ-UHFFFAOYSA-N 0.000 description 1
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 description 1
- 229910052782 aluminium Inorganic materials 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 235000001014 amino acid Nutrition 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 150000003863 ammonium salts Chemical class 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 1
- 235000010323 ascorbic acid Nutrition 0.000 description 1
- 229960005070 ascorbic acid Drugs 0.000 description 1
- 239000011668 ascorbic acid Substances 0.000 description 1
- 238000003705 background correction Methods 0.000 description 1
- SRSXLGNVWSONIS-UHFFFAOYSA-N benzenesulfonic acid Chemical compound OS(=O)(=O)C1=CC=CC=C1 SRSXLGNVWSONIS-UHFFFAOYSA-N 0.000 description 1
- 229940092714 benzenesulfonic acid Drugs 0.000 description 1
- 235000010233 benzoic acid Nutrition 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 150000001621 bismuth Chemical class 0.000 description 1
- 230000036772 blood pressure Effects 0.000 description 1
- KDYFGRWQOYBRFD-NUQCWPJISA-N butanedioic acid Chemical compound O[14C](=O)CC[14C](O)=O KDYFGRWQOYBRFD-NUQCWPJISA-N 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- BVKZGUZCCUSVTD-UHFFFAOYSA-N carbonic acid Chemical compound OC(O)=O BVKZGUZCCUSVTD-UHFFFAOYSA-N 0.000 description 1
- 238000001142 circular dichroism spectrum Methods 0.000 description 1
- 235000015165 citric acid Nutrition 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 235000001671 coumarin Nutrition 0.000 description 1
- 229960000956 coumarin Drugs 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 206010012601 diabetes mellitus Diseases 0.000 description 1
- ZBCBWPMODOFKDW-UHFFFAOYSA-N diethanolamine Chemical class OCCNCCO ZBCBWPMODOFKDW-UHFFFAOYSA-N 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 239000002552 dosage form Substances 0.000 description 1
- CCIVGXIOQKPBKL-UHFFFAOYSA-M ethanesulfonate Chemical compound CCS([O-])(=O)=O CCIVGXIOQKPBKL-UHFFFAOYSA-M 0.000 description 1
- 229930003944 flavone Natural products 0.000 description 1
- 150000002212 flavone derivatives Chemical class 0.000 description 1
- 235000011949 flavones Nutrition 0.000 description 1
- 239000011888 foil Substances 0.000 description 1
- 235000019253 formic acid Nutrition 0.000 description 1
- 229960002598 fumaric acid Drugs 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 230000004927 fusion Effects 0.000 description 1
- LRBQNJMCXXYXIU-QWKBTXIPSA-N gallotannic acid Chemical compound OC1=C(O)C(O)=CC(C(=O)OC=2C(=C(O)C=C(C=2)C(=O)OC[C@H]2[C@@H]([C@@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)[C@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)[C@@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)O2)OC(=O)C=2C=C(OC(=O)C=3C=C(O)C(O)=C(O)C=3)C(O)=C(O)C=2)O)=C1 LRBQNJMCXXYXIU-QWKBTXIPSA-N 0.000 description 1
- 229960005219 gentisic acid Drugs 0.000 description 1
- 239000000174 gluconic acid Substances 0.000 description 1
- 235000012208 gluconic acid Nutrition 0.000 description 1
- 229950006191 gluconic acid Drugs 0.000 description 1
- 229940116332 glucose oxidase Drugs 0.000 description 1
- 235000019420 glucose oxidase Nutrition 0.000 description 1
- 235000013922 glutamic acid Nutrition 0.000 description 1
- 239000004220 glutamic acid Substances 0.000 description 1
- 229940093915 gynecological organic acid Drugs 0.000 description 1
- 238000003919 heteronuclear multiple bond coherence Methods 0.000 description 1
- 238000001052 heteronuclear multiple bond coherence spectrum Methods 0.000 description 1
- 238000005570 heteronuclear single quantum coherence Methods 0.000 description 1
- 238000000990 heteronuclear single quantum coherence spectrum Methods 0.000 description 1
- 238000002114 high-resolution electrospray ionisation mass spectrometry Methods 0.000 description 1
- XMBWDFGMSWQBCA-UHFFFAOYSA-N hydrogen iodide Chemical compound I XMBWDFGMSWQBCA-UHFFFAOYSA-N 0.000 description 1
- 229940071870 hydroiodic acid Drugs 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- 230000003451 hyperinsulinaemic effect Effects 0.000 description 1
- 201000008980 hyperinsulinism Diseases 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 230000002452 interceptive effect Effects 0.000 description 1
- QXJSBBXBKPUZAA-UHFFFAOYSA-N isooleic acid Natural products CCCCCCCC=CCCCCCCCCC(O)=O QXJSBBXBKPUZAA-UHFFFAOYSA-N 0.000 description 1
- 239000004310 lactic acid Substances 0.000 description 1
- 235000014655 lactic acid Nutrition 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 229910003002 lithium salt Inorganic materials 0.000 description 1
- 159000000002 lithium salts Chemical class 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 159000000003 magnesium salts Chemical class 0.000 description 1
- VZCYOOQTPOCHFL-UPHRSURJSA-N maleic acid Chemical compound OC(=O)\C=C/C(O)=O VZCYOOQTPOCHFL-UPHRSURJSA-N 0.000 description 1
- 239000011976 maleic acid Substances 0.000 description 1
- 229960002510 mandelic acid Drugs 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 229940098779 methanesulfonic acid Drugs 0.000 description 1
- XELZGAJCZANUQH-UHFFFAOYSA-N methyl 1-acetylthieno[3,2-c]pyrazole-5-carboxylate Chemical compound CC(=O)N1N=CC2=C1C=C(C(=O)OC)S2 XELZGAJCZANUQH-UHFFFAOYSA-N 0.000 description 1
- 150000007522 mineralic acids Chemical class 0.000 description 1
- 229910017604 nitric acid Inorganic materials 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- ZQPPMHVWECSIRJ-KTKRTIGZSA-N oleic acid Chemical compound CCCCCCCC\C=C/CCCCCCCC(O)=O ZQPPMHVWECSIRJ-KTKRTIGZSA-N 0.000 description 1
- 229960002969 oleic acid Drugs 0.000 description 1
- 235000021313 oleic acid Nutrition 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 235000005985 organic acids Nutrition 0.000 description 1
- 235000006408 oxalic acid Nutrition 0.000 description 1
- 238000004806 packaging method and process Methods 0.000 description 1
- 238000012856 packing Methods 0.000 description 1
- WLJNZVDCPSBLRP-UHFFFAOYSA-N pamoic acid Chemical compound C1=CC=C2C(CC=3C4=CC=CC=C4C=C(C=3O)C(=O)O)=C(O)C(C(O)=O)=CC2=C1 WLJNZVDCPSBLRP-UHFFFAOYSA-N 0.000 description 1
- 239000011713 pantothenic acid Substances 0.000 description 1
- 229940055726 pantothenic acid Drugs 0.000 description 1
- 235000019161 pantothenic acid Nutrition 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 230000002093 peripheral effect Effects 0.000 description 1
- 108040007629 peroxidase activity proteins Proteins 0.000 description 1
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N phenol group Chemical group C1(=CC=CC=C1)O ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 1
- 150000002989 phenols Chemical class 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 239000008055 phosphate buffer solution Substances 0.000 description 1
- 235000013824 polyphenols Nutrition 0.000 description 1
- XAEFZNCEHLXOMS-UHFFFAOYSA-M potassium benzoate Chemical compound [K+].[O-]C(=O)C1=CC=CC=C1 XAEFZNCEHLXOMS-UHFFFAOYSA-M 0.000 description 1
- 125000001844 prenyl group Chemical group [H]C([*])([H])C([H])=C(C([H])([H])[H])C([H])([H])[H] 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 235000019260 propionic acid Nutrition 0.000 description 1
- IUVKMZGDUIUOCP-BTNSXGMBSA-N quinbolone Chemical compound O([C@H]1CC[C@H]2[C@H]3[C@@H]([C@]4(C=CC(=O)C=C4CC3)C)CC[C@@]21C)C1=CCCC1 IUVKMZGDUIUOCP-BTNSXGMBSA-N 0.000 description 1
- 238000010992 reflux Methods 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 229960004889 salicylic acid Drugs 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 159000000000 sodium salts Chemical class 0.000 description 1
- 150000001629 stilbenes Chemical class 0.000 description 1
- 235000021286 stilbenes Nutrition 0.000 description 1
- 239000011550 stock solution Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 235000015523 tannic acid Nutrition 0.000 description 1
- 229940033123 tannic acid Drugs 0.000 description 1
- 229920002258 tannic acid Polymers 0.000 description 1
- 235000002906 tartaric acid Nutrition 0.000 description 1
- 239000011975 tartaric acid Substances 0.000 description 1
- 229960001367 tartaric acid Drugs 0.000 description 1
- ISIJQEHRDSCQIU-UHFFFAOYSA-N tert-butyl 2,7-diazaspiro[4.5]decane-7-carboxylate Chemical compound C1N(C(=O)OC(C)(C)C)CCCC11CNCC1 ISIJQEHRDSCQIU-UHFFFAOYSA-N 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 238000002211 ultraviolet spectrum Methods 0.000 description 1
- VHBFFQKBGNRLFZ-UHFFFAOYSA-N vitamin p Natural products O1C2=CC=CC=C2C(=O)C=C1C1=CC=CC=C1 VHBFFQKBGNRLFZ-UHFFFAOYSA-N 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
- 150000003751 zinc Chemical class 0.000 description 1
Images
Classifications
-
- 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
-
- 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
- A61P3/06—Antihyperlipidemics
-
- 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
- A61P5/00—Drugs for disorders of the endocrine system
- A61P5/48—Drugs for disorders of the endocrine system of the pancreatic hormones
- A61P5/50—Drugs for disorders of the endocrine system of the pancreatic hormones for increasing or potentiating the activity of insulin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
-
- 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)
- Diabetes (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Animal Behavior & Ethology (AREA)
- Veterinary Medicine (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Pharmacology & Pharmacy (AREA)
- Life Sciences & Earth Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Endocrinology (AREA)
- Hematology (AREA)
- Obesity (AREA)
- Emergency Medicine (AREA)
- Cardiology (AREA)
- Heart & Thoracic Surgery (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses a flavonoid compound extracted from black mulberry plants, and a preparation method and application thereof. Specifically discloses a flavonoid compound I or pharmaceutically acceptable salt thereof and a preparation method of the flavonoid compound I. The preparation method of the flavonoid compound I comprises the steps of extraction, column chromatography and HPLC preparation. The flavonoid compound I can reduce the content of glucose in cell fluid, raise the level of adiponectin, and has small side effects.
Description
Technical Field
The invention relates to a flavonoid compound extracted from black mulberry plants, and a preparation method and application thereof.
Background
Along with the improvement of the social life level and the change of the mode, the type 2 diabetics are increased continuously, and the healthy life of people is seriously influenced. Among them, insulin resistance (Insulin resistance, IR) is one of the main causes of the onset of type 2 diabetes. It is mainly manifested in that the sensitivity of peripheral tissues such as liver, adipose tissue and the like to insulin is reduced, so that the organism competitively generates more insulin to maintain blood sugar level, hyperinsulinemia is generated, and further glycolipid metabolic disorder is caused, and metabolic diseases such as type 2 diabetes and the like are caused. Rosiglitazone, a drug for the treatment of diabetes, can improve IR resistance and increase insulin sensitivity, but also brings about a number of adverse effects.
Black mulberry (Morus nigra L.) is Moraceae (Moraceae) Morus (Morus) plant, and has effects of lowering blood sugar, blood lipid, blood pressure, and resisting virus, and the like, and the related researches report that black mulberry leaf and stem contain abundant active substances such as phenols, including flavonoids, stilbenes, DA adducts, etc.; alkaloid component, coumarin component, etc. Wherein the water-soluble and phenolic components have the function of reducing blood sugar, and the water-soluble alkaloid components are developed into mulberry twig total alkaloids tablets to be marketed at present.
Disclosure of Invention
The invention aims to solve the technical problem of providing a flavonoid compound extracted from black mulberry plants, and a preparation method and application thereof. The flavonoid compound can improve insulin resistance and increase insulin sensitivity.
The technical problems are solved by the following technical scheme.
The invention provides flavonoid compound I or pharmaceutically acceptable salt thereof,
the invention also provides a preparation method of the flavonoid compound I, which comprises the following steps:
(1) Extracting: extracting coarse powder of ramulus Mori with ethanol, and concentrating the extractive solution to obtain extract;
(2) Suspending the extract with water, performing AB-8 macroporous resin column chromatography, eluting with water, 40% ethanol and 95% ethanol respectively;
(3) Performing silica gel column chromatography on the eluent obtained by eluting with 95% ethanol, and performing gradient elution by using a mixed solution of petroleum ether and ethyl acetate, wherein the volume ratio of petroleum ether to ethyl acetate is 10:1,7:1,5:1,2:1,1:1,2:3, collecting the 16 th component;
(4) Subjecting the 16 th component to gel column chromatography, eluting with dichloromethane and methanol at a volume ratio of 1:1, and collecting the 4 th component;
(5) Subjecting the 4 th component to reverse phase silica gel chromatography, and carrying out gradient elution by using methanol and water, wherein the volume ratio of the methanol to the water is 50:50, 55:45, 60:40, 65:35, 70:30, 75:25, 80:20, 85:15, 90:10, 95:5,100: 0, collecting the 19 th component, concentrating to obtain a crude product of the flavonoid compound I;
(6) HPLC preparation: purifying the crude product by HPLC, wherein the stationary phase of the chromatographic column is C18 bonded silica gel, and the mobile phase is a mixed solution of acetonitrile and water; the volume ratio of acetonitrile to water is 60:40.
in one embodiment of the present invention, in the step (1), the ethanol is 85% to 95% ethanol, for example, 90% ethanol.
In one embodiment of the invention, in step (1), the extraction temperature is 75-90 ℃, for example 80 ℃.
In one embodiment of the present invention, in the step (1), the volume/mass ratio of the ethanol to the black mulberry shoot coarse powder is 5 to 10L/kg, for example 8L/kg.
In one embodiment of the present invention, in the step (1), the number of times of the extraction is 1 to 3, for example, 2.
In one embodiment of the invention, in step (1), the extraction time is 0.5 to 2 hours, for example 1 hour.
In a certain aspect of the present invention, in step (1), the extracting may further include the steps of: filtering the extractive solutions, mixing, and concentrating to obtain extract.
In one embodiment of the present invention, in the step (2), when the extract is suspended with water, the weight ratio of the extract to the water is 1:5 to 15, for example 1:10.
in one embodiment of the present invention, in the step (2), the water, 40% ethanol and 95% ethanol are used in an amount of 5 column volumes, respectively, when the AB-8 macroporous resin column chromatography is performed.
In one embodiment of the present invention, in the step (3), the eluent obtained by eluting with 95% ethanol is concentrated and dried, and then subjected to silica gel column chromatography.
In one embodiment of the present invention, in the step (3), the mixed solvent is used in an amount of 3 column volumes per gradient when the gradient elution is performed with a mixed solution of petroleum ether and ethyl acetate.
In one embodiment of the present invention, in step (3), the 16 th component is the eluent collected at 16 th column volumes. When the 16 th component is detected by silica gel thin layer chromatography, and the volume ratio of the developing agent to the methylene dichloride is 20:1, the Rf value can be 0.50.
In one embodiment of the present invention, in step (4), the 16 th component is concentrated under reduced pressure and then subjected to gel column chromatography.
In one embodiment of the present invention, in the step (4), the packing material of the gel column is hydroxypropyl sephadex LH-20.
In one embodiment of the present invention, in step (5), the 4 th component is concentrated and dried, and then subjected to reverse phase silica gel chromatography.
In one aspect of the present invention, in the step (5), the reverse phase silicse:Sup>A gel is ODS-C18, such as ODS-A-HG.
In one embodiment of the present invention, in step (5), the amount of the mixed solvent used for each gradient is 3 column volumes when the gradient elution is performed with methanol and water.
In one embodiment of the present invention, in the step (5), the 19 th component is eluted with a gradient of methanol and water, and the volume ratio of methanol to water is 95:5, using an amount of 2 column volumes of eluent collected. When the 19 th component is detected by silica gel thin layer chromatography, and the volume ratio of the developing agent to the methylene dichloride is 20:1, the Rf value can be 0.56.
In one embodiment of the invention, in the HPLC preparation, the filler particle size of the column is 5 to 10. Mu.m, for example 5. Mu.m; the length of the chromatographic column may be from 100 to 250mm, for example 250mm; the inner diameter of the column may be 9.4 to 30mm, for example 10mm.
In one embodiment of the present invention, the HPLC preparation method comprises the step of preparing the HPLC column with Agilent 1100.
In one embodiment of the invention, the HPLC preparation is carried out at a column temperature of 20 to 50℃for example at room temperature.
In one embodiment of the invention, the HPLC preparation has a detection wavelength of 200 to 400nm, for example 210nm.
In one embodiment of the invention, the flow rate of the mobile phase in the HPLC preparation is 1-15 mL/min, such as 3mL/min.
In one embodiment of the invention, the HPLC preparation is performed with a sample volume of 10 to 1000. Mu.L, for example 200. Mu.L.
The present invention also provides a pharmaceutical composition comprising: flavonoid I or pharmaceutically acceptable salt thereof, and at least one pharmaceutical adjuvant.
The pharmaceutical composition may also include a thiazolidinedione component.
The thiazolidinedione component may be rosiglitazone or pioglitazone hydrochloride.
The pharmaceutical excipients can be pharmaceutical excipients conventionally used in the art, preferably sucrose, gelatin, glycerol, pregelatinized starch, microcrystalline cellulose, lactose or inorganic calcium salts.
The pharmaceutical dosage form of the pharmaceutical composition can be a tablet, a capsule, a granule, a powder or a liquid preparation.
The pharmaceutical composition may be prepared using methods known to those skilled in the art.
The pharmaceutical composition can be used in combination with other drugs for treating type 2 diabetes.
The invention also provides application of the flavonoid compound I, the pharmaceutically acceptable salt or the pharmaceutical composition thereof in preparing medicaments for preventing and treating insulin resistance and diseases caused by the insulin resistance. The diseases include insulin resistance and metabolic diseases caused by the insulin resistance. The metabolic diseases preferably include type 2 diabetes, hyperlipidemia and cardiovascular diseases.
In such applications, preferably, the flavonoid or pharmaceutically acceptable salt thereof is the sole active ingredient.
The flavonoid compound I, a pharmaceutically acceptable salt or a pharmaceutical composition thereof may be administered to a subject by any suitable route, preferably orally, intravenously or topically, more preferably orally.
The term "pharmaceutically acceptable salt" refers to salts of the compounds of the present invention prepared with relatively non-toxic, pharmaceutically acceptable acids or bases. When the compounds of the present invention contain relatively acidic functional groups, base addition salts can be obtained by contacting a prototype of such compounds with a sufficient amount of a pharmaceutically acceptable base in pure solution or in a suitable inert solvent. Pharmaceutically acceptable base addition salts include, but are not limited to: lithium salt, sodium salt, potassium salt, calcium salt, aluminum salt, magnesium salt, zinc salt, bismuth salt, ammonium salt, diethanolamine salt. When relatively basic functional groups are present in the compounds of the present invention, the acid addition salts may be obtained by contacting a prototype of such compounds with a sufficient amount of a pharmaceutically acceptable acid in pure solution or in a suitable inert solvent. The pharmaceutically acceptable acids include inorganic acids including, but not limited to: hydrochloric acid, hydrobromic acid, hydroiodic acid, nitric acid, carbonic acid, phosphoric acid, phosphorous acid, sulfuric acid, and the like. The pharmaceutically acceptable acid includes organic acids including, but not limited to: acetic acid, propionic acid, oxalic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, salicylic acid, tartaric acid, methanesulfonic acid, isonicotinic acid, acidic citric acid, oleic acid, tannic acid, pantothenic acid, hydrogen tartrate, ascorbic acid, gentisic acid, fumaric acid, gluconic acid, sugar acid, formic acid, ethanesulfonic acid, pamoic acid (i.e., 4' -methylene-bis (3-hydroxy-2-naphthoic acid)), amino acids (e.g., glutamic acid, arginine), and the like. When the compounds of the present invention contain relatively acidic and relatively basic functional groups, they can be converted into base addition salts or acid addition salts. See, for example, berge et al, "Pharmaceutical Salts", journal of Pharmaceutical Science 66:1-19 (1977), or Handbook of Pharmaceutical Salts: properties, selection, and Use (P.Heinrich Stahl and Camille G.Wermuth, ed., wiley-VCH, 2002).
The term "treatment" refers to therapeutic therapy. When specific conditions are involved, treatment refers to: (1) alleviating a disease or one or more biological manifestations of a disorder, (2) interfering with (a) one or more points in a biological cascade that results in or causes a disorder or (b) one or more biological manifestations of a disorder, (3) ameliorating one or more symptoms, effects, or side effects associated with a disorder, or one or more symptoms, effects, or side effects associated with a disorder or treatment thereof, or (4) slowing the progression of a disorder or one or more biological manifestations of a disorder.
The above preferred conditions can be arbitrarily combined on the basis of not deviating from the common knowledge in the art, and thus, each preferred embodiment of the present invention can be obtained.
The reagents and materials used in the present invention are commercially available.
The invention has the positive progress effects that: the invention discovers flavonoid compounds shown in the formula I, which can reduce the content of glucose in cell fluid, raise the level of adiponectin and have small side effects.
Drawings
FIG. 1 shows the CD spectrum of flavonoid I (JASCO Corp., J-810. Solvent used: chromatographic grade acetonitrile; test concentration: 0.03 mg/mL) obtained in example 1;
FIG. 2 shows a chiral resolution of flavonoid I obtained in example 1 at 246nm (chiral column: waters Trefoil) TM CEL 2.5 μm Columns (150X 3.0mm,2.5 μm); chromatographic conditions: acetonitrile and water are used as mobile phases, and the volume ratio of the acetonitrile to the water is 60:40);
FIG. 3 shows HSQC spectra (acetone-d) of flavonoid I obtained in example 1 6 ,600MHz)；
FIG. 4 shows HMBC spectra (acetone-d) of flavonoid I obtained in example 1 6 ,600MHz)。
Detailed Description
The invention is further illustrated by means of the following examples, which are not intended to limit the scope of the invention. The experimental methods, in which specific conditions are not noted in the following examples, were selected according to conventional methods and conditions, or according to the commercial specifications.
EXAMPLE 1 preparation of flavonoid I
1. Extracting: extracting dried coarse powder of ramulus Mori with 90% ethanol under reflux, wherein the amount of 90% ethanol is 8 (v/m, L/kg) times of the amount of the medicinal materials, and extracting twice for 1 hr each time. Filtering the extractive solutions, mixing, and concentrating to obtain extract.
2. Column chromatography: suspending the extract with 10 times of water, performing AB-8 macroporous resin column chromatography, eluting with water, 40% ethanol and 95% ethanol respectively, and eluting with 5 times of column volume. Concentrating and drying the eluent obtained by eluting with 95% ethanol, performing silica gel column chromatography, and using petroleum ether with the volume of three times of column volume: the mixed solutions of ethyl acetate (10:1, 7:1,5:1,2:1,1:1, 2:3) are respectively subjected to gradient elution, and are identified by a silica gel thin layer plate, and the mixed solutions are combined with the same Rf value to obtain 23 components.
Component 16 (i.e., eluent collected when the amount of eluent was 16 th column volumes, i.e., eluent collected when the amount of mixed solvent (petroleum ether: ethyl acetate=2:1) was 2 column volumes) was collected, and the developing agent was methylene chloride by thin-layer chromatography detection of silica gel: methanol=20:1, rf value=0.50; concentrating under reduced pressure, subjecting to gel column (Sephadex LH-20), GE Healthcare Bio-Sciences AB, uppsala, sweden) chromatography, eluting with dichloromethane: methanol at 1:1, sequentially collecting with 15mL test tubes, identifying with silica gel thin layer plate, combining test tubes according to Rf value to obtain 7 components, and collecting fourth component (silica gel thin layer chromatography detection, developing solvent is dichloromethane: methanol=20:1); after concentration and drying, the mixture was subjected to reverse phase silicse:Sup>A gel ODS-C18 (ODS-se:Sup>A-HG, YMC co., ltd., japan) chromatography, and eluted with se:Sup>A gradient of methanol and water (50:50, 55:45, 60:40, 65:35, 70:30, 75:25, 80:20, 85:15, 90:10, 95:5, 100:0), the mixed solvent amount used for each gradient was 3 column volumes, and the mixture was collected in sequence for each column volume, and was identified by silicse:Sup>A gel thin layer plates, rf values were the same, and 22 fractions were obtained in total, fraction 19 (i.e., the eluate collected when the amount of the mixed solvent (methanol: water=95:5) was 2 column volumes) was collected, and rf=0.56 when the developer was methylene chloride: methanol=20:1 was detected by silicse:Sup>A gel thin layer chromatography, and the mixture was concentrated and dried to obtain se:Sup>A crude product of the flavonoid compound I.
Hplc preparation: purifying the crude flavonoid compound I by HPLC (chromatographic column model: agilent 1100,5 μm, 10X1250 mm; sample injection amount: 200 μl; column temperature: room temperature; stationary phase: C18 bonded silica gel; mobile phase: 60% acetonitrile aqueous solution, detection wavelength 210nm, flow rate 3mL/min; preparation liquid phase: LC3050N type high performance liquid chromatograph) to obtain flavonoid compound I: (E) -5- (2, 6-dimethylhepta-1, 5-dien-1-yl) -3,4,8,10-tetrahydroxy-11- (3-methybut-2-en-1-yl) isochromeno [4,3-b ] chrome-7 (5H) -one.
EXAMPLE 2 structural identification of flavonoid I
A pale yellow oil; HR ESI MS m/z 503.2039[M-H] - Molecular formula C 30 H 32 O 7 The method comprises the steps of carrying out a first treatment on the surface of the The UV spectrum (methanol as solvent) shows maximum absorption at 264.06 and 384.06nm, and is presumed to be flavonoid. CD (c 0.03mg/mL, ACN) values were near 0 (see FIG. 1), and HPLC analysis (FIG. 2) using chiral columns gave two closer peaks, presumably a pair of racemates.
1 H-NMR(600MHz,acetone-d 6 (ii) the spectrum shows the signal delta of the hydroxy group at the C-5 position of a flavone H 12.93 (1H, s, OH-5); two ortho-coupled aromatic proton signals 7.31 (1 h, d, j=6.8 hz, h-6 '), 7.02 (1 h, d, j=6.8 hz, h-5'); an aromatic proton signal 6.32 (1H, s, H-6); a set of prenyl signals 5.33 (1H, br t, J=6.8 Hz, H-10), 3.62 (1H, br dd, J=14.4, 6.8Hz, H-9 a), 3.52 (1H, br dd, J=14.4, 6.8Hz, H-9 b), 1.84 (3H, br s, H-13), 1.68 (3H, br s, H-12); in addition, there are three methine signals 6.34 (1H, d, J=7.6 Hz, H-14), 5.54 (1H, d, J=8.8 Hz, H-15), 4.98 (1H, t, J=6.4 Hz, H-20); two methylene signals 2.01 (2H, m, H-18), 1.99 (2H, m, H-19); and three methyl signals 1.99 (3H, s, H-17), 1.50 (3H, br s, H-23), 1.48 (3H, br s, H-22).
13 C-NMR(150MHz,acetone-d 6 ) The spectra showed 30 carbons, 15 of which were δc176.8 (C-4), δc 161.9 (C-7), δ0c161.4 (C-5), δ1c154.8 (C-8 a), δ2c149.7 (C-4 '), δ3c149.4 (C-2), δ4c142.0 (C-3'), δ5c132.9 (C-3), δ6c117.0 (C-1 '), δ7c123.9 (C-2'), δ8c115.5 (C-6 '), δ9c115.7 (C-5'), δc107.4 (C-8), δ0c106.5 (C-4 a), δ1c99.2 (C-6); 5 carbon signals on 1 isopentenyl 22.6 (C-9), 123.9 (C-10), 132.3 (C-11), 26.2 (C-12), 18.5 (C-13). In combination with HSQC (FIG. 3), three methines were found to correspond to δ2c70.3 (C-14), δc122.5 (C-15), δc124.9 (C-20), respectively; two methylene groups correspond to δc40.5 (C-18), δc27.1 (C-19), respectively; three methyl groups correspond to δc25.9 (C-22), δc18.0 (C-23), δc17.5 (C-17), respectively; two quaternary carbons 131.5 (C-21), 132.2 (C-16). In combination with HMBC (FIG. 4), H-9 is associated with C-7, confirming that an isopentenyl group is attached at the C-8 position; h-14 is associated with C-3, C-1', C-3', C-15 and C-16, indicating that a six membered ring is present between the B ring and the C ring; h-17 is associated with C-15, C-18, H-19 is associated with C-18, H-20 is related to C-19 and C-23, and H-23 is related to C-20 and C-21. C-22-related indicates the presence of a side chain (E) -2,6-dimethylocta-2,6-diene at the C-14 position. The planar structure of the compound is determined.
EXAMPLE 3 insulin sensitization Studies with flavonoid I
1. Instrument for measuring and controlling the intensity of light
Enzyme-labeled instrument: bioTek, EPOCH; CO 2 Incubator (Thermo 3111), microscope (olympus CX 23), ultra clean bench (Su Xin YJ-840/YJ-1340), pipette (Eppendorf).
2. Material
3T3-L1 cells: mouse preadipocytes derived from a cell bank of the national academy of sciences; DMEM (Gibco, C11995500 CP); fetal bovine serum (Gibco, 2110875 CP); anti-anti (Lifetechnologies, 15240-112); PBS, pH7.4 (Gibco, 10010-500 BT); trypsin-EDTA (0.25%) (Gibco, 25200-056); IBMX (Sigma, I7018); dexamethasone (Sigma, D4902); insulin (Lifetechnologies, 12585-014); induction culture solution: 1X conventional culture solution; 0.5mM IBMX;1 μm Dexamethasone;2 μg/mL Insulin; preserving at 4 ℃ for standby. Dexamethasone stock:25mg of DEX was dissolved in 1mL of absolute ethanol (M.W = 392.46 g/mol) and stored at 4℃for further use as 64. Mu.M stock solution. IBMX stock:110mg of IBMX was dissolved in 1mL of DMSO (M.W =222.2 g/mol) and stored at-20℃for further use after packaging. Mouse Adiponectin ELISA Kit: biyun, PA002. Cell culture dish: coning,430167; cell culture flask: corning,430639; cell culture plate: costar,3596, 3599; glucose test cassette: nanjing is built, F006-1-1.
Sang Genchun F (Sanggenol F) is prepared by a laboratory, and the preparation method refers to the literature: xu Liangjin two national medicinal plants have chemical composition and biological activity research [ D ]. Shanghai, china medical industry institute, 2018.
3. Experimental step (1) glucose content determination
(1) Preadipocyte differentiation:
cell culture: mouse preadipocytes 3T3-L1 were routinely cultured in DMEM medium (containing 1X antibiotics) with 10% FBS.
Inducing differentiation: after 3T3-L1 is cultured until fusion, induction is carried out according to the following induction differentiation scheme, and the specific process is as follows:
day0: the conventional culture solution is changed into an induction culture solution, and the culture is carried out for 48 hours;
day2: collecting cells, changing the induction culture solution into a culture solution containing insulin, and culturing for 48 hours;
day4: the culture was continued by changing the DMEM medium containing 10% FBS, changing the medium every 48 hours, and adding the drug for intervention on day 6.
(2) Cell supernatant glucose content assay:
the enzyme-labeled instrument is preheated for more than 30min, the wavelength is regulated to 505nm, and the distilled water is zeroed.
Sample assay (the following reagents were added sequentially to a 1.5mL EP tube):
i sample: compound I, sangggenol F, rosiglitazone
ii Reagent one: phosphate buffer solution 100mmol/L, p-hydroxybenzoic acid 16.5mmol/L iii Mixing the following reagents: phosphate buffer 100mmol/L, glucose oxidase 105KU/L, peroxidase 1.2KU/L
Mixing, keeping the temperature at 37 ℃ for 10min, and reading absorbance at 505nm wavelength. The absorbance values of the blank, standard and assay tubes are designated A1, A2 and A3, respectively.
The formula: glucose content (μmol/mL) =c standard X (A3-A1)/(A2-A1)
Standard tube concentration, 5.55. Mu. Mol/mL.
(2) Determination of adiponectin content
(1) The number of pre-coated strips required for one experiment was calculated and determined, the required strips were removed and placed in a 96 well frame, temporarily without strips, and the strips were returned to the aluminum foil bag for sealing and stored at 4 ℃.
(2) The standard substance is prepared and a standard curve is drawn in each experiment, and a background correction hole, namely a blank hole, is suggested to be arranged, and the arrangement method is that only TMB solution and stop solution are added into the hole.
(3) Samples or standards of different concentrations were added to the corresponding wells at 100 μl/well, and the reaction wells were sealed with a sealing plate membrane (transparent) and incubated at room temperature for 120 minutes.
(4) The plate is washed 5 times, and finally the plate is placed on thick water absorbing paper for beating. 100. Mu.L/well of biotinylated antibody was added (note: this biotinylated antibody was already pre-formulated and used directly without further dilution). The reaction wells were sealed with a sealing plate membrane (transparent) and incubated at room temperature for 60 minutes.
(5) The plate is washed 5 times, and finally the plate is placed on thick water absorbing paper for beating.
(6) 100. Mu.L/well of horseradish peroxidase-labeled strepitavidine was added (note: this horseradish peroxidase-labeled strepitavidine was pre-formulated and used as such without further dilution). The reaction wells were sealed with a sealing plate membrane (white), and incubated at room temperature for 20 minutes in the dark. At low room temperature (below 25 ℃), it is necessary to extend the incubation time appropriately.
(7) The plate is washed 5 times, and finally the plate is placed on thick water absorbing paper for beating.
(8) 100 mu L/well of TMB solution as a color developing agent is added, the reaction well is sealed by a sealing plate film (white), and the reaction well is incubated for 15-20 minutes at room temperature and in a dark place. At low room temperature, the incubation time needs to be prolonged appropriately, and the standard can be incubated until the standard has a very significant color change, and if the sample concentration is high enough, the color change is significant.
(9) The stop solution was added at 50. Mu.L/well, and the A450 value was measured immediately after mixing.
The values of the complex pores are usually valid within 20% of the difference, and the average value of the complex pores can be used as the measured value.
(3) 3T3-L1 preadipocyte differentiation
(1) Preadipocyte differentiation:
the procedure was the same as in the glucose assay described above.
(2) Measurement of ability to promote adipocyte differentiation:
after dissolution with isopropanol, the OD490 nm was measured by an enzyme-labeled instrument and the reading was recorded.
4. Experimental results
TABLE 1 effect of compounds on glucose content in insulin resistant 3T3-L1 adipocyte model.
(n＝3,`x±SD)
Note that: low dose: compound I and Sanggenol F were 10 μm and rosiglitazone was 1 μm;
high dose: compound I and Sanggenol F were 30 μm and rosiglitazone was 10 μm;
p < 0.05, < p < 0.01 compared to model group; compared to the normal group, #p < 0.01.
Table 2: effect of the compounds on adiponectin content in the insulin resistant 3T3-L1 adipocyte model.
(n＝3,`x±SD)
Note that: low dose: compound I and Sanggenol F were 10 μm and rosiglitazone was 1 μm;
high dose: compound I and Sanggenol F were 30 μm and rosiglitazone was 10 μm;
p < 0.05 compared to the model group, # p < 0.01 compared to the normal group.
The results in tables 1 and 2 show that the flavonoid I of the present invention has significantly better ability to lower the glucose content and raise the adiponectin level in the cell fluid at the low dose of 10 mu M and the high dose of 30 mu M than Sanggenol F. The ability of flavonoid I to reduce glucose content in cell fluid and raise adiponectin level at low dose concentration of 10 μm is equivalent to that of positive medicine rosiglitazone.
Table 3: effect of compounds on differentiation of insulin resistant 3T3-L1 adipocyte model into lipid.
Examination of the effect of 3T3-L1 preadipocyte induced differentiation shows that flavonoid I has no effect of promoting the differentiation of the adipocytes at the concentration of 10 mu M, and rosiglitazone has the effect of promoting the differentiation of the adipocytes into lipid at the concentration of 1 mu M (side effect), which shows that the flavonoid I has less side effect at the concentration of 10 mu M than that of the rosiglitazone at the concentration of 1 mu M.
In conclusion, the insulin sensitization activity of the flavonoid compound I is superior to that of Sanggenol F, and the flavonoid compound I has small side effect and potential development value.
Claims (10)
1. Flavonoid I or pharmaceutically acceptable salt thereof,
2. a process for the preparation of flavonoids I according to claim 1, characterized in that it comprises the following steps:
(1) Extracting: extracting coarse powder of ramulus Mori with ethanol, and concentrating the extractive solution to obtain extract;
(2) Suspending the extract with water, performing AB-8 macroporous resin column chromatography, eluting with water, 40% ethanol and 95% ethanol respectively;
(3) Performing silica gel column chromatography on the eluent obtained by eluting with 95% ethanol, and performing gradient elution by using a mixed solution of petroleum ether and ethyl acetate, wherein the volume ratio of petroleum ether to ethyl acetate is 10:1,7:1,5:1,2:1,1:1,2:3, collecting the 16 th component;
(4) Subjecting the 16 th component to gel column chromatography, eluting with dichloromethane and methanol at a volume ratio of 1:1, and collecting the 4 th component;
(5) Subjecting the 4 th component to reverse phase silica gel chromatography, and carrying out gradient elution by using methanol and water, wherein the volume ratio of the methanol to the water is 50:50, 55:45, 60:40, 65:35, 70:30, 75:25, 80:20, 85:15, 90:10, 95:5,100: 0, collecting the 19 th component, concentrating to obtain a crude product of the flavonoid compound I;
(6) HPLC preparation: purifying the crude flavonoid compound I by HPLC, wherein the stationary phase of the chromatographic column is C18 bonded silica gel, and the mobile phase is a mixed solution of acetonitrile and water; the volume ratio of acetonitrile to water is 60:40.
3. the process for the preparation of flavonoids I according to claim 2, wherein the process satisfies one or more of the following conditions:
(1) In the step (1), the ethanol is 85% -95% ethanol;
(2) In the step (1), the extraction temperature is 75-90 ℃;
(3) In the step (1), the volume-mass ratio of the ethanol to the black mulberry stem and branch coarse powder is 5-10L/kg;
(4) In the step (1), the extraction times are 1-3 times;
(5) In the step (1), the extraction time is 0.5-2 hours;
(6) In the step (1), the extracting further includes the steps of: filtering the extractive solutions, mixing, and concentrating to obtain extract;
(7) In the step (2), when the extract is suspended by water, the weight ratio of the extract to the water is 1:5 to 15;
(8) In the step (2), when AB-8 macroporous resin column chromatography is carried out, the dosage of water, 40% ethanol and 95% ethanol is 5 times of column volume respectively;
(9) In the step (3), concentrating and drying the eluent obtained by eluting with 95% ethanol, and performing silica gel column chromatography;
(10) In the step (3), when the mixed solution of petroleum ether and ethyl acetate is used for gradient elution, the amount of the mixed solvent used for each gradient is 3 times of the volume of the column;
(11) In the step (3), the 16 th component is eluent collected when the 16 th column volume is multiplied;
(12) In the step (4), the 16 th component is concentrated under reduced pressure and then subjected to gel column chromatography;
(13) In the step (4), the filler of the gel column is hydroxypropyl sephadex LH-20;
(14) In the step (5), the 4 th component is concentrated and dried and then subjected to reverse phase silica gel chromatography;
(15) In the step (5), the reverse phase silica gel is ODS-C18;
(16) In the step (5), when methanol and water are used for gradient elution, the amount of the mixed solvent used for each gradient is 3 times of the volume of the column;
(17) In the step (5), the 19 th component is eluted with methanol and water in a gradient, and the volume ratio of methanol to water is 95:5, using an amount of 2 column volumes of eluent collected;
(18) In the HPLC preparation, the granularity of the filler of the chromatographic column is 5-10 mu m; the length of the chromatographic column can be 100-250 mm; the inner diameter of the chromatographic column can be 9.4-30 mm;
(19) In the HPLC preparation, the chromatographic column is Agilent 1100;
(20) In the HPLC preparation, the column temperature of the chromatographic column is 20-50 ℃;
(21) In the HPLC preparation, the detection wavelength is 200-400 nm;
(22) In the HPLC preparation, the flow rate of the mobile phase is 1-15 mL/min;
(23) In the HPLC preparation, the sample injection volume is 10-1000 mu L.
4. A process for the preparation of flavonoid I according to claim 3, which process satisfies one or more of the following conditions:
(1) In the step (1), the ethanol is 90% ethanol;
(2) In the step (1), the extraction temperature is 80 ℃;
(3) In the step (1), the volume-mass ratio of the ethanol to the black mulberry stem and branch coarse powder is 8L/kg;
(4) In the step (1), the number of times of extraction is 2;
(5) In the step (1), the extraction time is 1 hour;
(6) In the step (2), when the extract is suspended by water, the weight ratio of the extract to the water is 1:10;
(7) In the step (5), the reverse phase silicse:Sup>A gel is ODS-A-HG;
(8) In the HPLC preparation, the filler granularity of the chromatographic column is 5 mu m; the length of the chromatographic column may be 250mm; the inner diameter of the chromatographic column may be 10mm;
(9) In the HPLC preparation, the column temperature of the chromatographic column is room temperature;
(10) In the HPLC preparation, the detection wavelength is 210nm;
(11) In the HPLC preparation, the flow rate of the mobile phase is 3mL/min;
(12) In the HPLC preparation, the sample volume was 200. Mu.L.
5. A pharmaceutical composition comprising a flavonoid I or a pharmaceutically acceptable salt thereof according to claim 1, and at least one pharmaceutical excipient.
6. The pharmaceutical composition of claim 5, wherein the pharmaceutical composition satisfies one or more of the following conditions:
(1) The pharmaceutical composition further comprises a thiazolidinedione component; the thiazolidinedione component can be rosiglitazone or pioglitazone hydrochloride;
(2) The pharmaceutical excipients are sucrose, gelatin, glycerol, pregelatinized starch, microcrystalline cellulose, lactose or inorganic calcium salts;
(3) The medicinal formulation of the medicinal composition is tablets, capsules, granules, powder or liquid preparations.
7. Use of a flavonoid I, a pharmaceutically acceptable salt thereof or a pharmaceutical composition according to claim 5 or 6 according to claim 1 for the preparation of a medicament for the prevention and treatment of insulin resistance and diseases caused thereby.
8. The use according to claim 7, wherein the disease is insulin resistance and metabolic diseases caused thereby; the metabolic diseases preferably include type 2 diabetes, hyperlipidemia and cardiovascular diseases.
9. The use according to claim 7, wherein the flavonoid I or a pharmaceutically acceptable salt thereof is the sole active ingredient.
10. Use according to claim 7 or 8, wherein the flavonoid I, a pharmaceutically acceptable salt thereof or the pharmaceutical composition is administered orally, intravenously or topically, e.g. orally.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111644417.8A CN116410201A (en) | 2021-12-29 | 2021-12-29 | Flavonoid compound extracted from black mulberry plants and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111644417.8A CN116410201A (en) | 2021-12-29 | 2021-12-29 | Flavonoid compound extracted from black mulberry plants and preparation method and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN116410201A true CN116410201A (en) | 2023-07-11 |
Family
ID=87058125
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202111644417.8A Pending CN116410201A (en) | 2021-12-29 | 2021-12-29 | Flavonoid compound extracted from black mulberry plants and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN116410201A (en) |
-
2021
- 2021-12-29 CN CN202111644417.8A patent/CN116410201A/en active Pending
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JP6302102B2 (en) | A compound isolated from MONASCUS PURPUREUS, its preparation and use | |
| WO2021164155A1 (en) | Cryptotanshinone derivative, preparation method therefor and application thereof in resisting neuroinflammation and neuroprotection | |
| WO2022257995A1 (en) | Cryptotanshinone derivative and preparation method therefor and application thereof in lowering lipid and resisting obesity | |
| US10093695B2 (en) | Sterol derivative, preparation method therefor and use thereof | |
| CN112915096B (en) | Pharmaceutical application of echinocystic acid-28-O-beta-D-glucoside | |
| CN101502506B (en) | Medical use of 3-(3,4-dihydroxyphenyl)-acrylic acid 2-(3,4-dihydroxyphenyl)-ethyl ester | |
| CN111150740A (en) | Lipid-lowering active ingredient and lipid-lowering composition in panax japonicus and application of lipid-lowering active ingredient and lipid-lowering composition | |
| CN114262294B (en) | Phenyl isoquinoline alkaloid compound and preparation method and application thereof | |
| CN116410201A (en) | Flavonoid compound extracted from black mulberry plants and preparation method and application thereof | |
| CN111166735B (en) | Use of bis- (2-ethylheptyl) phthalate for inhibiting fat accumulation | |
| CN110437149B (en) | Natural naphthyl isoquinoline compound with antitumor activity, and composition and application thereof | |
| Wang et al. | Icaritin inhibits endometrial carcinoma cells by suppressing O-GlcNAcylation of FOXC1 | |
| CN109876022B (en) | Black fungus blood fat reducing active part and preparation method and application thereof | |
| CN113057955A (en) | Medicine for inhibiting stearoyl-CoA desaturase 1 | |
| CN108358947B (en) | Caged xanthone compound and preparation method and application thereof | |
| JP2005503381A (en) | Sesquiterpenoid derivatives with adipocyte differentiation inhibitory action | |
| CN116947794B (en) | Eucalyptus type sesquiterpenoids rearranged by four-ring system, preparation method and application thereof, pharmaceutical composition and application thereof | |
| CN109206392B (en) | Coumarin compound and preparation method and application thereof | |
| CN118047788A (en) | Isopentenyl phenolic compound, and preparation method and application thereof | |
| CN105796680B (en) | Willow leaf meadow sweet stem branch resisting rheumatoid disease active component and its preparation method and application | |
| CN105524135B (en) | The preparation method and its application in preparation of anti-tumor drugs of straw berry tomato lactone | |
| CN114591282B (en) | Pyranone compound separated from moldavica dragonhead as well as preparation method and application thereof | |
| CN114533719B (en) | Application of abietane diterpenoid compound in preparation of anti-inflammatory drugs | |
| WO2021063076A1 (en) | Application of albiziae cortex lignan compound in improving steatosis | |
| CN115073400B (en) | Preparation method and application of guaiacolide sesquiterpene compound with hypoglycemic activity 1, 10-split ring |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |