CN103936812B - Lupinane type triterpene compound and pharmaceutical composition thereof are applied with it - Google Patents
Lupinane type triterpene compound and pharmaceutical composition thereof are applied with it Download PDFInfo
- Publication number
- CN103936812B CN103936812B CN201410177372.1A CN201410177372A CN103936812B CN 103936812 B CN103936812 B CN 103936812B CN 201410177372 A CN201410177372 A CN 201410177372A CN 103936812 B CN103936812 B CN 103936812B
- Authority
- CN
- China
- Prior art keywords
- compound
- cell
- pharmaceutical composition
- type triterpene
- triterpene compound
- 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.)
- Active
Links
- VZEGBIWAIFLVJH-NXEZZACHSA-N (1r,9ar)-1-methyl-2,3,4,6,7,8,9,9a-octahydro-1h-quinolizine Chemical compound C1CCC[C@@H]2[C@H](C)CCCN21 VZEGBIWAIFLVJH-NXEZZACHSA-N 0.000 title claims abstract description 10
- 239000008194 pharmaceutical composition Substances 0.000 title claims abstract description 10
- -1 triterpene compound Chemical class 0.000 title abstract description 9
- 239000003814 drug Substances 0.000 claims abstract description 25
- 238000002360 preparation method Methods 0.000 claims abstract description 18
- 229940125904 compound 1 Drugs 0.000 claims abstract description 17
- 229940125782 compound 2 Drugs 0.000 claims abstract description 17
- 239000003937 drug carrier Substances 0.000 claims abstract description 5
- 150000003648 triterpenes Chemical class 0.000 claims abstract description 5
- 206010012601 diabetes mellitus Diseases 0.000 claims description 7
- 230000002265 prevention Effects 0.000 claims description 4
- 150000003839 salts Chemical class 0.000 abstract description 9
- 230000000694 effects Effects 0.000 abstract description 7
- 239000000203 mixture Substances 0.000 abstract description 6
- 230000003178 anti-diabetic effect Effects 0.000 abstract description 4
- 239000003472 antidiabetic agent Substances 0.000 abstract description 3
- 239000000470 constituent Substances 0.000 abstract 1
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 23
- 210000004027 cell Anatomy 0.000 description 22
- 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 18
- 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 16
- 239000008103 glucose Substances 0.000 description 16
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 15
- 229940079593 drug Drugs 0.000 description 13
- 102000004877 Insulin Human genes 0.000 description 12
- 108090001061 Insulin Proteins 0.000 description 12
- 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 12
- 229960003957 dexamethasone Drugs 0.000 description 12
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 11
- 239000013642 negative control Substances 0.000 description 10
- 235000015097 nutrients Nutrition 0.000 description 10
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 9
- 150000001875 compounds Chemical class 0.000 description 9
- 229940093499 ethyl acetate Drugs 0.000 description 9
- 229940125396 insulin Drugs 0.000 description 9
- 239000000741 silica gel Substances 0.000 description 9
- 229910002027 silica gel Inorganic materials 0.000 description 9
- 229960001866 silicon dioxide Drugs 0.000 description 9
- 206010022489 Insulin Resistance Diseases 0.000 description 8
- 230000003833 cell viability Effects 0.000 description 8
- 210000001789 adipocyte Anatomy 0.000 description 6
- 239000007788 liquid Substances 0.000 description 6
- 239000012930 cell culture fluid Substances 0.000 description 5
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 4
- 241000208832 Viburnum Species 0.000 description 4
- 239000002775 capsule Substances 0.000 description 4
- 238000000605 extraction Methods 0.000 description 4
- 239000003981 vehicle Substances 0.000 description 4
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 3
- 244000132059 Carica parviflora Species 0.000 description 3
- 235000014653 Carica parviflora Nutrition 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
- MUBZPKHOEPUJKR-UHFFFAOYSA-N Oxalic acid Chemical compound OC(=O)C(O)=O MUBZPKHOEPUJKR-UHFFFAOYSA-N 0.000 description 3
- 241001529246 Platymiscium Species 0.000 description 3
- 238000010609 cell counting kit-8 assay Methods 0.000 description 3
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 3
- 230000000857 drug effect Effects 0.000 description 3
- 229960004756 ethanol Drugs 0.000 description 3
- 239000012737 fresh medium Substances 0.000 description 3
- 239000006187 pill Substances 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- 241000208828 Caprifoliaceae Species 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- 244000307537 Viburnum odoratissimum Species 0.000 description 2
- 235000019013 Viburnum opulus Nutrition 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 238000004113 cell culture Methods 0.000 description 2
- 239000006285 cell suspension Substances 0.000 description 2
- 230000008021 deposition Effects 0.000 description 2
- 238000004821 distillation Methods 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 239000000706 filtrate Substances 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 230000006698 induction Effects 0.000 description 2
- HQKMJHAJHXVSDF-UHFFFAOYSA-L magnesium stearate Chemical compound [Mg+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O HQKMJHAJHXVSDF-UHFFFAOYSA-L 0.000 description 2
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 2
- 238000000034 method Methods 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 239000003960 organic solvent Substances 0.000 description 2
- 238000005453 pelletization Methods 0.000 description 2
- 239000000546 pharmaceutical excipient Substances 0.000 description 2
- 239000000049 pigment Substances 0.000 description 2
- 229940093429 polyethylene glycol 6000 Drugs 0.000 description 2
- 239000013641 positive control Substances 0.000 description 2
- OVARTBFNCCXQKS-UHFFFAOYSA-N propan-2-one;hydrate Chemical compound O.CC(C)=O OVARTBFNCCXQKS-UHFFFAOYSA-N 0.000 description 2
- 238000010298 pulverizing process Methods 0.000 description 2
- 238000010898 silica gel chromatography Methods 0.000 description 2
- 241000894007 species Species 0.000 description 2
- 230000003595 spectral effect Effects 0.000 description 2
- 239000007921 spray Substances 0.000 description 2
- 239000003826 tablet Substances 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 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
- 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
- 241001465754 Metazoa Species 0.000 description 1
- 241001529936 Murinae Species 0.000 description 1
- 108010019160 Pancreatin Proteins 0.000 description 1
- 229920003171 Poly (ethylene oxide) Polymers 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 235000015401 Viburnum lentago Nutrition 0.000 description 1
- 235000013249 Viburnum odoratissimum Nutrition 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000001464 adherent effect Effects 0.000 description 1
- 230000002293 adipogenic effect Effects 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 239000012752 auxiliary agent Substances 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 244000309466 calf Species 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 235000015165 citric acid Nutrition 0.000 description 1
- 238000004440 column chromatography Methods 0.000 description 1
- 238000009833 condensation Methods 0.000 description 1
- 230000005494 condensation Effects 0.000 description 1
- 230000024835 cytogamy Effects 0.000 description 1
- 229960000935 dehydrated alcohol Drugs 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- 229930004069 diterpene Natural products 0.000 description 1
- 150000004141 diterpene derivatives Chemical class 0.000 description 1
- 239000012154 double-distilled water Substances 0.000 description 1
- 239000008298 dragée Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 238000013210 evaluation model Methods 0.000 description 1
- 229930003944 flavone Natural products 0.000 description 1
- 150000002213 flavones Chemical class 0.000 description 1
- 235000011949 flavones Nutrition 0.000 description 1
- 235000011194 food seasoning agent Nutrition 0.000 description 1
- 235000019253 formic acid Nutrition 0.000 description 1
- 239000007903 gelatin capsule Substances 0.000 description 1
- 125000002791 glucosyl group Chemical group C1([C@H](O)[C@@H](O)[C@H](O)[C@H](O1)CO)* 0.000 description 1
- 206010073071 hepatocellular carcinoma Diseases 0.000 description 1
- WGCNASOHLSPBMP-UHFFFAOYSA-N hydroxyacetaldehyde Natural products OCC=O WGCNASOHLSPBMP-UHFFFAOYSA-N 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 238000009413 insulation Methods 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 208000030208 low-grade fever Diseases 0.000 description 1
- 235000019359 magnesium stearate Nutrition 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 235000012054 meals Nutrition 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 235000006408 oxalic acid Nutrition 0.000 description 1
- 229940055695 pancreatin Drugs 0.000 description 1
- 244000045947 parasite Species 0.000 description 1
- 239000000825 pharmaceutical preparation Substances 0.000 description 1
- 210000000229 preadipocyte Anatomy 0.000 description 1
- 238000002953 preparative HPLC Methods 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 230000008439 repair process Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 229960003271 rosiglitazone maleate Drugs 0.000 description 1
- SUFUKZSWUHZXAV-BTJKTKAUSA-N rosiglitazone maleate Chemical compound [H+].[H+].[O-]C(=O)\C=C/C([O-])=O.C=1C=CC=NC=1N(C)CCOC(C=C1)=CC=C1CC1SC(=O)NC1=O SUFUKZSWUHZXAV-BTJKTKAUSA-N 0.000 description 1
- 229920006395 saturated elastomer Polymers 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 238000002791 soaking Methods 0.000 description 1
- 239000012453 solvate Substances 0.000 description 1
- 210000000952 spleen Anatomy 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 238000005728 strengthening Methods 0.000 description 1
- 239000007940 sugar coated tablet Substances 0.000 description 1
- 239000006188 syrup Substances 0.000 description 1
- 235000020357 syrup Nutrition 0.000 description 1
- 229940095064 tartrate Drugs 0.000 description 1
- 229940098465 tincture Drugs 0.000 description 1
Landscapes
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
Thering is provided lupinane type triterpene compound 1 and 2 or its pharmaceutical salts, take it as the pharmaceutical composition of the pharmaceutically acceptable carrier composition of effective constituent and at least one, its preparation method, and it is preparing the application in antidiabetic medicine.1 and 2 is lupinane type triterpene compound, has significant external insulin-sensitizing activity, active strong, has more advantage as antidiabetic medicine.
Description
Technical field:
The invention belongs to technical field of pharmaceuticals, specifically, the lupinane type triterpene compound with anti-diabetic activity is related to, its preparation method, with the pharmaceutical composition that this compound is activeconstituents, and they treat the application in the medicine of diabetes in preparation.
Background technology:
Viburnum (Viburnum) is under the jurisdiction of Caprifoliaceae (Caprifoliaceae), and the whole world about has 200 kinds, is distributed in temperate zone and subtropical zone, Asia and South America kind more.China about has 74 kinds, is distributed widely in national each provinces and regions, with west and south most species.This platymiscium is the important medicinal and ornamental plant monoid of a class.The numerous species of Viburnum plant has pharmaceutical use, and wherein folk medicinal plants about has 43 kinds.Be mainly used in clearing heat and detoxicating, dispel rheumatism, strengthening the spleen to promote digestion, expelling parasite, kobadrin etc.Just because of there being many kinds all to have long-term Medicinal basis in this platymiscium, various countries successively expand a large amount of research work, especially in study of pharmacy to this platymiscium.Successively find from different Viburnum plants as diterpene, triterpene, lignanoid and flavones isoreactivity composition.Triterpene compound structure type is enriched, and has biological activity widely.There are no the report about lupinane type triterpene compound and activity thereof in prior art.
Summary of the invention:
The object of the invention is to provide two lupinane type triterpene compounds that separation obtains from sweet viburnum (ViburnumodoratissimumKer-Gawl.), with the pharmaceutical composition that it is activeconstituents, its preparation method, they are particularly preparing the application in antidiabetic medicine in pharmacy.
In order to realize above-mentioned purpose of the present invention, the invention provides following technical scheme:
Lupinane type triterpene compound 1 and 2 shown in following structural formula or its pharmaceutical salts,
16β-hydroxy-lup-20(29)-en-3-on-28-oicacidR
1＝β-OH
26a-hydroxy-lup-20(29)-en-3-on-28-oicacidR
2＝a-OH。
As described in lupinane type triterpene compound 1 and 2 or its pharmaceutical salts, wherein said pharmaceutical salts refers to pharmacy acceptable salt, with organic acid as tartrate or citric acid or formic acid or oxalic acid, or the salt formed with mineral acid example hydrochloric acid or sulfuric acid or phosphoric acid.
For preventing or treat a pharmaceutical composition for diabetes, it comprises described compound 1 or 2 or its pharmaceutical salts and the pharmaceutically acceptable carrier of at least one.
The preparation method of compound 1 and 2, with 100% acetone or alcohol or methanol solvate, cold soaking or thermal backflow lixiviate coral branch, leaf meal obtain total medicinal extract, and total medicinal extract must described compound after various positive reversed phase column chromatography and preparation are separated with half preparative HPLC.
As as described in preparation method, get coral branch, leaf, dry, with 70% acetone water at room temperature lixiviate three times after pulverizing, each 2 days, united extraction liquid, after underpressure distillation removing majority of organic solvent, hold over night, the pigment of filtering deposition, filtrate uses sherwood oil and extraction into ethyl acetate three times respectively, merge after concentrating and obtain ethyl acetate portion, used 95% dissolve with ethanol, MCI post is used to decolour, silicagel column on gained sample silica gel mixed sample after decolouring merging evaporate to dryness, the sherwood oil of (1:0-0:1) and ethyl acetate rush post in varing proportions, detect with TLC, merge identical fraction, obtain seven parts, wherein second section is through sherwood oil-chlorofonn-ethylacetate silica gel column chromatography repeatedly, use Rp-18 reversed-phase column 45%-80%MeOH:H again
2o is divided into three components, and component 2 is through SephadexLH-20 post, and chloroform: methyl alcohol=1:1 obtains compound 1, Part III silica gel mixed sample is crossed silicagel column and is divided into three components, and component 3 after SephadexLH-20 post, then obtains 2 through Rp-18 separation.
Compound 1 and 2 or the application of its pharmaceutical salts in preparation prevention or treatment diabetes medicament.
With the application of pharmaceutical composition in preparation prevention or treatment diabetes medicament that compound 1 and 2 or its pharmaceutical salts form for activeconstituents.
When the compounds of this invention 1 and 2 is used as medicine, directly can uses, or use with the form of pharmaceutical composition.This pharmaceutical composition contains 0.1 – 99%, and be preferably the compounds of this invention of 0.5 – 90%, all the other are acceptable on pharmacology, pharmaceutically acceptable carrier of and inertia nontoxic to humans and animals or vehicle.
Described pharmaceutically acceptable carrier or vehicle are that one or more are selected from solid, semisolid and liquid diluent, extra-fill material and pharmaceutical preparation assistant agent.Described effective extract or efficient part are used with the form of per weight dose.Medicine of the present invention can oral administration and mouth spray two kinds of form administrations.
Oral its solid available or liquid preparation, as pulvis, tablet, sugar coated tablet, capsule, tincture, syrup, pill etc.
Mouth spray can with its solid or liquid preparation.
Medicine of the present invention can be used for treating diabetes.
Embodiment:
Further illustrate essentiality content of the present invention with embodiments of the invention below, but do not limit the present invention with this.
Embodiment 1:
The preparation of compound 1 and 2:
Dry coral branch, leaf 4kg, with 70% acetone water (each 30 liters) at room temperature lixiviate three times after pulverizing, each 2 days.United extraction liquid, after underpressure distillation removing majority of organic solvent, hold over night, the pigment of filtering deposition, filtrate uses sherwood oil and extraction into ethyl acetate three times respectively, merge concentrated after obtain ethyl acetate portion and be about 305g.Ethyl acetate portion is used 95% dissolve with ethanol, MCI post is used to decolour, decolouring obtains sample after merging evaporate to dryness and is about 268g, silicagel column on silica gel mixed sample, the sherwood oil of (1:0-0:1) and ethyl acetate rush post in varing proportions, detect with TLC, merge identical fraction, obtain seven parts.Wherein second section is through sherwood oil-chlorofonn-ethylacetate silica gel column chromatography repeatedly, then uses Rp-18 reversed-phase column (45%-80%MeOH:H
2o) be divided into three components, component 2 obtains compound 1 (523mg) through SephadexLH-20 post (chloroform: methyl alcohol=1:1).Part III silica gel mixed sample is crossed silicagel column and is divided into three components, and component 3 after SephadexLH-20 post, then obtains 2 (565mg) through Rp-18 separation.
Compound structure is resolved:
NMR spectral data (the CDCl of table-1. compounds 1
3)
NMR spectral data (the CD of table-2. compounds 2
3cOCD
3)
Embodiment 2:
The external insulin-sensitizing activity evaluation of compound 1,2:
Medicine and reagent: DMEM substratum, foetal calf serum (FBS) and pancreatin are Gibco product, Glucose estimation kit (lot number 2012041) is Changchun Hui Li Bioisystech Co., Ltd product, CellCountingKit-8 (CCK-8) test kit is that Japanese colleague Co., Ltd. produces, dexamethasone, Regular Insulin and DMSO are Sigma product, and rosiglitazone maleate is provided by institute for drug control, Guangzhou.
Clone and cell cultures: adopt human hepatoma cell line HepG2, adipocyte that murine preadipocyte cell system 3T3-L1 induction becomes is as being evaluation model.The nutrient solution of HepG2 cell is with the addition of 10%FBS, DMEM containing 2g/L glucose, and the nutrient solution of 3T3-L1 PECTORAL LIMB SKELETON is the high pool DMEM that with the addition of 10%FBS.Cell culture condition: 37 DEG C, saturated humidity, 5% CO2gas incubator.
Drug effect: the HepG2 cell of (1) collected by trypsinisation logarithmic phase, is mixed with 5 × 10 with cell culture fluid
4the single cell suspension of individual/ml, is inoculated in 96 orifice plates by 100 μ L/ holes, cultivates 24h and makes cell completely adherent.Discard old nutrient solution, add the fresh medium that 100 μ L include 1 μM of dexamethasone and different concns medicine to be tested, continue cultivation and make drug treating time be 48h.The each drug level of this experiment sets three multiple holes, and set simultaneously not celliferous blank well, non-drug containing and dexamethasone negative control hole and containing 1 μM of dexamethasone but not containing the Positive control wells of other drug.(2) 3T3-L1 PECTORAL LIMB SKELETON cell culture fluid is mixed with 5 × 10
4the single cell suspension of individual/ml, is inoculated in 96 orifice plates by 100 μ L/ holes, is placed in incubator and makes cytogamy, changes a nutrient solution every other day.Cell is carried out adipogenic induction to break up its differentiation of 8 angels and become adipocyte, namely in the cell culture fluid containing 1 μM of dexamethasone, 0.5mM3-isobutyl--1-methyl xanthine, 10 μ g/ml Regular Insulin 2 days, containing in the cell culture fluid of 10 μ g/ml Regular Insulin 2 days, in the cell culture fluid of not drug containing 4 days, change a nutrient solution every other day.The nutrient solution changing the adipocyte after having broken up is the fresh medium that 100 μ L include 1 μM of dexamethasone and different concns medicine to be tested, continues cultivation and makes drug treating time be 48h.The each drug level of this experiment sets three multiple holes, and set simultaneously not celliferous blank well, non-drug containing and dexamethasone negative control hole and containing 1 μM of dexamethasone but not containing the Positive control wells of other drug.
Cell sugar consumption measures: take out 96 orifice plates, 10 μ L nutrient solutions are taken out in every hole, adopt the Glucose estimation kit (lot number 2012041) of Changchun Hui Li Bioisystech Co., Ltd, measure the amount remaining glucose in nutrient solution according to test kit operation instructions.According to accurately recording the concentration of glucose and the concentration of fresh medium glucose in the nutrient solution of every hole, calculate the glucose of every porocyte 48h internal consumption, and cell under drug effect is relative to the glucose consumption value RGC (%) of negative control porocyte.
Cell viability measures: take out 10 μ L nutrient solutions for 96 orifice plates after glucose assays, the CCK-8 of 10 μ L is added in every hole, take out after cultivating 1-2h in incubator, measure the absorption value A in every hole at microplate reader 450nm place, this optical density value and number of viable cells positive correlation.Can show that the cell under different concns drug effect to calculate the cell viability under different pharmaceutical concentration relative to the vigor CV (%) of control wells cell according to A value.Cell viability %=100 × (A
determinand-A
blank)/(A
negative-A
blank).
Pharmaceutical activity is evaluated: the cell viability (CV) in versus glucose consumption (RGC)/every hole in glucose consumption ability (GCA)=every hole of Board Lot viable cell.
Result: (1) HepG2 cell (insulin resistant) vigor under 1 μM of dexamethasone exists is 95% of negative control (insulin sensitivity), and glucose consumption is 82.1% (table 3) of negative control; And 3T3-L1 adipocyte (insulin resistant) vigor under 1 μM of dexamethasone exists is 100% of negative control (insulin sensitivity), glucose consumption is then 84.6% (table 4) of negative control.Illustrate that the vigor of the dexamethasone of 1 μM on cell is without impact, but the picked-up of cell to glucose can be reduced more than 15%, illustrate that dexamethasone inducing cell produces the model establishment of insulin resistant.(2) on insulin resistant HepG2 cell model, the vigor of 1 and 2 pair of cell of working concentration is without obviously suppressing (CV>90%), but cell is taken in glucose significantly increase, even exceed the negative control of insulin sensitivity, show good insulin-sensitizing effect (table 3); (3) on the 3T3-L1 adipocyte model of insulin resistant, 1 of working concentration and 2 on the vigor of cell without impact (CV>97.5%), but make the absorption ability of cell to glucose return to the level of insulin responsive cells (negative control), show the effect (table 4) of well repairing insulin sensitivity.
Table 3. compound 1 and 2 strengthens the insulin sensitivity means standard deviation (n=3) of insulin resistant HepG2 cell
Table 4. compound 1 and 2 repairs the insulin sensitivity means standard deviation (n=3) of insulin resistant 3T3-L1 adipocyte
Compare with the negative control of insulin sensitivity, data have significant difference:
▲p<0.05and
▲ ▲p<0.01;
Compare with the model comparison of insulin resistant, data have significant difference: * P<0.05, * * P<0.01and***P<0.001.
Conclusion: compound 1 and 2 has the effect of obvious external enhancing insulin sensitivity.
Embodiment 3:
Obtain compound 2 by embodiment 1, add vehicle in itself and excipient weight than the ratio of 1:1, pelletizing press sheet.
Embodiment 4:
Obtain compound 2 by embodiment 1, add vehicle in itself and excipient weight than the ratio of 1:2, pelletizing press sheet.
Embodiment 5:
Obtain compound 2 by embodiment 1, capsule preparations method makes capsule routinely.
Embodiment 6:
Obtain compound 2 by embodiment 1, then make tablet as follows:
Embodiment 7:
Capsule: compound 2100mg
Starch is appropriate
Magnesium Stearate proper quantity
Preparation method: mix 2 with auxiliary agent, sieves, Homogeneous phase mixing in suitable container, and the mixture obtained is loaded hard gelatin capsule.
Embodiment 8:
Preparation method: at every turn add a kind of composition under stirring in the double distilled water of proper volume, separates until completely dark, and then adds another kind of composition.After adding water to 2ml, this solution is filtered on sterilizing filter, to load in bottle and to separate according to suitable dosage.
Embodiment 9:
Dripping pill: compound 1 or 21g
Polyethylene glycol 6000 9g
Method for making: the preparation of compound 1 or 2 and polyethylene glycol 6000 fused solution: take 1 or 2 by above-mentioned recipe quantity, add appropriate dehydrated alcohol, after low-grade fever is dissolved, add (60 DEG C of water bath heat preservations) in the polyoxyethylene glycol fused solution of recipe quantity, be uniformly mixed, till ethanol is waved to the greatest extent, be statically placed in 60 DEG C of water-baths and be incubated 30 minutes, treat that bubble eliminates, then the above-mentioned mixing fused solution eliminating bubble is proceeded in liquid storage cylinder, under the condition of insulation 80-85 DEG C, control to drip speed, instill dropwise in phlegma, complete Deng condensation, incline phlegma, collect dripping pill, drop is clean and remove the phlegma on ball with filter paper, place in silica gel drier or seasoning.
Claims (2)
1. the application of the lupinane type triterpene compound 1 and 2 shown in following structural formula in preparation prevention or treatment diabetes medicament,
2. comprise the compound 1 described in claim 1 or/and 2 and the pharmaceutically acceptable carrier of at least one pharmaceutical composition preparation prevention or treatment diabetes medicament in application.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410177372.1A CN103936812B (en) | 2014-04-29 | 2014-04-29 | Lupinane type triterpene compound and pharmaceutical composition thereof are applied with it |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410177372.1A CN103936812B (en) | 2014-04-29 | 2014-04-29 | Lupinane type triterpene compound and pharmaceutical composition thereof are applied with it |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103936812A CN103936812A (en) | 2014-07-23 |
| CN103936812B true CN103936812B (en) | 2015-11-11 |
Family
ID=51184716
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410177372.1A Active CN103936812B (en) | 2014-04-29 | 2014-04-29 | Lupinane type triterpene compound and pharmaceutical composition thereof are applied with it |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103936812B (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106397623A (en) * | 2016-10-12 | 2017-02-15 | 重庆大学 | Rapid preparation method of polygonum multiflorum decolorizing polysaccharide |
| CN111000849A (en) * | 2019-12-19 | 2020-04-14 | 淮阴师范学院 | Application and product of lupeol DYC-6 |
-
2014
- 2014-04-29 CN CN201410177372.1A patent/CN103936812B/en active Active
Non-Patent Citations (3)
| Title |
|---|
| Chemical Studies on Viburnum awabuki K. KOCH;MASANORI KUROYANAGI et al.，;《Chem. Pharm. Bull.》;19861231;第34卷(第10期);第4014页chart 2以及第4016页第2段 * |
| Cytotoxic lupane-，secolupane-，and oleanane-type triterpenes from Viburnum awabuki;ALI A. EL-GAMAL;《Natural Product Research》;20080215;第22卷(第3期);第192页第1段以及第194页第3段 * |
| 忍冬属植物三萜皂苷类化学成分研究进展;陈雨 等;《中草药》;20130628;第44卷(第12期);第1679-1686页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103936812A (en) | 2014-07-23 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Liu et al. | An in vivo and in vitro assessment of the anti-inflammatory, antinociceptive, and immunomodulatory activities of Clematis terniflora DC. extract, participation of aurantiamide acetate | |
| Saleem et al. | Hepatoprotective effect of aqueous methanolic extract of Rumex dentatus in paracetamol-induced hepatotoxicity in mice | |
| CN108743583B (en) | Application of sesquiterpene lactone compound in centipeda minima in preparation of antiviral drug | |
| CN104370871B (en) | The mouth diphenylene ketone oxide class separated from Swertia punicea Hemsl. and the application of suppression hepatitis B virus | |
| CN105111269A (en) | Novel limonin compound as well as preparation method and medical application thereof | |
| KR20130107408A (en) | Composition for inhibiting hepatic stellate cells activation | |
| CN101480422A (en) | Tibetan oriental wormwood extract as well as preparation method and use thereof | |
| US20130231469A1 (en) | Method for preparing albiflorin and paeoniflorin | |
| CN102641317A (en) | Golden wave extract and application thereof in preparation of antidiabetic agent | |
| CN101933966B (en) | Active site composition of isatis roots, as well as preparation method and application thereof | |
| CN103936812B (en) | Lupinane type triterpene compound and pharmaceutical composition thereof are applied with it | |
| CN109232491A (en) | The Preparation method and use of benzofuran compounds in a kind of Herba Serissae | |
| US11458180B2 (en) | Environmentally-friendly preparation method of composition containing anti-inflammatory active ingredients and use of composition | |
| CN101824014A (en) | Compounds with anti-tumor activity in chloranthus japonicus as well as effective parts and purpose thereof | |
| CN104382968A (en) | Method for extracting common andrographis paniculata total lactone, pharmaceutical composition of andrographis paniculata total lactone and use of pharmaceutical composition | |
| CN103288615A (en) | Monocyclic phloroglucinol compounds and pharmaceutical composition and application thereof | |
| CN105287610A (en) | Application of forsythiaside I and preparation method thereof | |
| CN103239435B (en) | Preparation method of gynura divaricata total caffeoylquinic acid and application in antihyperglycemic agent or health-care product | |
| CN103285082B (en) | Medicament composition for treating gynecological inflammation | |
| Mulholland et al. | Triterpenoid acids and lactones from the leaves of Fadogia tetraquetra var. tetraquetra (Rubiaceae) | |
| CN102188497B (en) | Preparation method of total alkali of corydalis tuber, and corydalis tuber preparation and use thereof | |
| CN101912453B (en) | Medicinal composition with anti-inflammatory and analgesic effects and preparation method and application thereof | |
| CN106177035B (en) | Preparation method and application of effective rosa chinensis flower extract with blood sugar reducing and anticancer functions | |
| CN104961719A (en) | Nature product juglandoid having bacterial inhibition activity, and preparation method thereof | |
| CN105395808B (en) | It is a kind of for treating the synergism medicine composition of diabetes |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| OL01 | Intention to license declared | ||
| OL01 | Intention to license declared |