CN104098632A - Iridoid glycoside compound, and preparation method and application thereof - Google Patents
Iridoid glycoside compound, and preparation method and application thereof Download PDFInfo
- Publication number
- CN104098632A CN104098632A CN201310145395.XA CN201310145395A CN104098632A CN 104098632 A CN104098632 A CN 104098632A CN 201310145395 A CN201310145395 A CN 201310145395A CN 104098632 A CN104098632 A CN 104098632A
- Authority
- CN
- China
- Prior art keywords
- methanol
- preparation
- cut
- water
- 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.)
- Pending
Links
Landscapes
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses an iridoid glycoside compound as shown in a formula I and a preparation method and application thereof. The preparation method comprises the following steps: (1) respectively extracting a Reduning injection with ethyl acetate and n-butanol so as to obtain n-butanol extract; (2) subjecting the n-butanol extract to HP-20 macroporous adsorption resin column chromatographic separation and ethanol-water gradient elution so as to obtain a 25-35% ethanol elution part; and (3) successively subjecting the 25-35% ethanol elution part to silica gel column chromatographic separation, ODS column chromatographic separation, HW-40 column chromatographic separation and preparative liquid phase HPLC separation so as to obtain the compound. The compound provided by the invention can be used for treating hand-foot-and-mouth disease.
Description
Technical field
The present invention relates to medical technical field, particularly a kind of iridoid glycoside compound extracting and its preparation method and application from existing Chinese patent medicine preparation.
Background technology
Reduning injection (the accurate word Z20050217 of traditional Chinese medicines) is former Chinese medicine two kind new medicines of Kangyuan Pharmaceutical Co., Ltd., Jiangsu Prov's independent research, and its prescription is sweet wormwood, Japanese Honeysuckle, cape jasmine, and auxiliary material is Polysorbate 80.Reduning injection is widely used in the clinical treatment of flu, influenza, cough, upper respiratory tract infection due to affection of exogenous wind-heat, and its effect is rapid, effect is remarkable.
Researchist has found new chemical composition in Reduning injection, and finds this compound equal stable existence in the Reduning injection of each batch.Known through By consulting literatures, this is the new chemical composition of finding in Reduning injection first and identifying, and retrieval finds that this compound is new compound through Scifinder scholar.
Summary of the invention
The present invention prepares to extract and a kind ofly has bioactive iridoid glycoside compounds from Reduning injection, and its application in preparation treatment hand foot mouth disease medicine is provided.
Specifically, the invention provides a kind of iridoid glycoside compounds, its structure is suc as formula shown in I:
Above-claimed cpd called after of the present invention: 2 '-O-is trans-coffee acyl gardoside (2 '-O-trans-caffeoylgardoside).
The present invention also provides the preparation method of above-claimed cpd, comprises the steps:
(1) get Reduning injection finished product, by isopyknic ethyl acetate and propyl carbinol, extract respectively, concentrating under reduced pressure obtains acetic acid ethyl ester extract, n-butyl alcohol extract and water layer;
(2) n-butyl alcohol extract of getting step (1) is through the separation of HP-20 macroporous adsorbent resin column chromatography, and alcohol-water gradient elution, obtains water elution position, 25～35% alcohol elutions, 95% alcohol elution;
(3) 25～35% alcohol elutions of getting step (2) are through silica gel column chromatography separation, use chloroform-methanol gradient elution, collect the cut E that chloroform-methanol ratio is 9: 1, cut E is separated through ODS column chromatography, use methanol-water gradient elution, collect the cut E-2 that methanol-water ratio is 1: 1, cut E-2 is separated through HW-40 column chromatography, use methanol-water gradient elution, collect the cut E-2-3 that methanol-water ratio is 6: 4, cut E-2-3 is separated through preparation liquid phase HPLC, obtains the compounds of this invention.
In above-mentioned preparation method, the described preparative liquid chromatography of step (3), the methanol-water that the ratio of take is 5: 5 is moving phase, detects wavelength and be 254 and 325nm, flow velocity 8mL/min, the retention time in preparation liquid phase is 38.8min.
Contriver by physico-chemical property and the Modern spectroscopy section of learning to do (UV, IR, MS,
1h-NMR,
13c-NMR and 2D-NMR) the separated compound obtaining has been carried out to Structural Identification, be confirmed that it is structure suc as formula the iridoid glycoside compound shown in I.
Another object of the present invention is to provide the application of compound shown in a kind of formula I in preparation treatment hand foot mouth disease medicine.Contriver finds that the compounds of this invention has certain restraining effect to hand-foot-mouth disease EV 71 virus.
A further object of the present invention is to provide a kind of pharmaceutical composition, comprises compound shown in above-mentioned formula I.
The present invention also provides a kind of pharmaceutical composition for the treatment of hand foot mouth disease, contains above-mentioned formula I compound and one or more pharmaceutically acceptable carriers for the treatment of significant quantity.
Accompanying drawing explanation
Fig. 1 is compound
1h-NMR spectrum;
Fig. 2 is the local spectrum of amplifying of the HMBC of compound;
Fig. 3 is main HMBC (→) and COSY (-) relevant information of compound;
Fig. 4 is compound
13c-NMR spectrum;
Fig. 5 is compound
1h-
1h COSY spectrum;
Fig. 6 is the hsqc spectrum of compound;
Embodiment
In order to make those skilled in the art understand better technical scheme of the present invention, below in conjunction with specific embodiment, the present invention is described in further detail.
The preparation of embodiment 1 the compounds of this invention
(1) get Reduning injection finished product 700g, with isopyknic ethyl acetate and propyl carbinol, extract respectively three times, concentrating under reduced pressure obtains acetic acid ethyl ester extract, n-butyl alcohol extract and water layer;
(2) n-butyl alcohol extract of getting step (1) is through the separation of HP-20 macroporous adsorbent resin column chromatography, and ethanol water gradient elution, obtains water elution position, 30% alcohol elution, 95% alcohol elution;
(3) 30% alcohol elution of getting step (2) is through silica gel column chromatography separation, use chloroform-methanol gradient elution, collect the cut E7.7g that chloroform-methanol ratio is 9: 1, cut E is separated through ODS column chromatography, use methanol-water gradient elution, collect the cut E-23.2g that methanol-water ratio is 1: 1, cut E-2 is separated through HW-40 column chromatography, use methanol-water gradient elution, collect the cut E-2-3299.0mg that methanol-water ratio is 6: 4, cut E-2-3 is separated through preparation liquid phase HPLC, the methanol-water that the ratio of take is 5: 5 is moving phase, detect wavelength and be 254 and 325nm, flow velocity 8mL/min, retention time at preparation liquid phase warp is 38.8min, separating obtained solution is dry, obtain the compounds of this invention 12.6mg.
The Structural Identification of embodiment 2 the compounds of this invention
Faint yellow gluey thing, Molisch reacting positive,
(c0.4, MeOH).ESI-MS (positive) provides m/z559[M+Na]
+and m/z1095[2M+Na]
+; ESI-MS (negative) provides m/z535[M-H]
-and m/z1071[2M-H]
-, prompting compound molecular weight is 536.HR-ESI-MS provides m/z559.1641[M+Na]
+(calculated value is 559.1635), deterministic adduct molecule formula C
25h
28o
13, calculating degree of unsaturation is 12.
The compounds of this invention
1h-NMR (400MHz, in CD
3oD) (Fig. 1) collection of illustrative plates, has shown the feature hydrogen signal of 1 coffee acyl, 1 glucosyl group and gardoside aglycon.Comprehensive above analysis, finally composes the connection site of each structure fragment of this compound to determine by HMBC.The connection site of each structure fragment can be composed and be determined by HMBC.HMBC amplifies in spectrum (Fig. 2), the distant relation peak of H-1 (δ 5.49)/C-1 ' (δ 97.9) and H-1 ' (δ 4.88)/C-1 (δ 96.4), and prompting glucosyl group is replaced in the C-1 position of aglycon; " (δ 168.2) have distant relation, and prompting coffee acyl is connected to the C-2 " position of glucosyl group for glycosyl H-2 ' (δ 4.80) and carbonyl C-1.Comprehensive above analysis, by the compounds of this invention be accredited as 2 '-O-trans-coffee acyl gardoside.
Comprehensive HSQC and HMBC spectrum information (Fig. 3), carried out belonging to (in Table 1) to whole carbon signals of compound and hydrogen signal.Through SciFinder Scholar network retrieval, do not find relevant report, show that this compound is a new iridoid glycoside compounds.
The nuclear magnetic data of table 1 compound (deuterated methanol,
1h-NMR400MHz,
13c NMR100MHz)
Embodiment 3 the compounds of this invention In Vitro Anti hand-foot-mouth disease EV 71 virus drug testings
1. material
1.1 strain hand-foot-mouth disease EV 71 virus, the laboratory preservation of going down to posterity.
1.2 cell model monkey-kidney cells are Vero, the laboratory preservation of going down to posterity.Culture condition: DMEM+10% foetal calf serum, 37 ℃, 5%CO
2.
2. principle and method
The cytotoxicity of 2.1 medicines detects
Adopted
(Invitrogen) toxic action of test kit detection of drugs to cell.
Experimental principle:
be a kind of oxidation-reduction indicator, can produce absorbancy according to metabolic activity and change and fluorescent signal.
soluble in water, its oxidised form enters cell and produces measurable fluorescence and colour-change by cyclophorase reduction, for quantitative analysis and the vitro cytotoxicity research of cytoactive and cell proliferation.This mensuration is that the cell based on having metabolic activity converts reagent to the ability of fluorescence and colorimetric indicator, and impaired and non-activity cell has lower natural metabolic activity, and corresponding signal is lower.Therefore fluorescent signal is strong and weak, can reflect the height of cytoactive.
Method steps: Vero cell is inoculated in 96 porocyte culture plates, standby after cell attachment.With cell maintenance medium (DMEM+5% serum) by medicine from 6 gradients of 2 times of continuous 3 times of gradient dilutions of initial concentration, every concentration gradient single hole detects.Dosing is cultivated after 48h, adds
hatch 2h, fluoroscopic examination for 37 ℃
reduction situation, exciting light 570nm, utilizing emitted light 595nm.
The inhibition test of cytoactive (%)=(sample well-blank)/(cell contrast-blank) * 100%2.2 medicines to EV71 virus
Containing after the EV71 vero cells infection of reporter gene GFP, cells infected meeting expressing green fluorescent protein, by express the cell number of GFP green fluorescence at fluorescence microscopy Microscopic observation, just can reflect the propagation situation of EV71 virus.
Method steps:
Vero cell is inoculated in 96 porocyte culture plates, standby after cell attachment.Medicine is 6 gradients of continuous 3 times of gradient dilutions from 4 times of test concentrations the highest; The medicine having diluted is added in hand-hole, after 4h, add viral supernatant liquor to infect, be placed in 37 ℃ of cell culture incubators and cultivate 24h, under fluorescent microscope, take pictures, fluorocyte is counted.
Experiment is established without medicine control wells (not adding medicine hole after virus infection), positive drug control wells (Guanidinium hydrochloride GuHCl).
Inhibiting rate (%)=(without medicine control wells-sample well)/without medicine control wells * 100%
3. result
3.1 drug samples detect the toxicity of cell and EV71 are suppressed to active and detect
Drug sample dilution solvent for use, the highest concentration of ordinary dissolution, the highest test concentrations, CC
50, EC
50and SI (selectivity index) is as shown in table 2.
Table 2 experimental result
4. conclusion
The compounds of this invention shows overt toxicity, and examines under a microscope most cells change and justify and have cracking, when (100 μ M), also has overt toxicity, and its SI value is 11.4, shows that it has certain restraining effect to hand-foot-mouth disease EV 71 virus.
The above is only the preferred embodiment of the present invention; it should be pointed out that for those skilled in the art, under the premise without departing from the principles of the invention; can also make some improvements and modifications, these improvements and modifications also should be considered as protection scope of the present invention.
Claims (6)
1. an iridoid glycoside compounds, its structure is suc as formula shown in I:
2. the preparation method of compound described in claim 1, is characterized in that, comprises the following steps:
(1) get Reduning injection finished product, by isopyknic ethyl acetate and propyl carbinol, extract respectively, concentrating under reduced pressure obtains acetic acid ethyl ester extract, n-butyl alcohol extract and water layer;
(2) n-butyl alcohol extract of getting step (1) is through the separation of HP-20 macroporous adsorbent resin column chromatography, and alcohol-water gradient elution, obtains water elution position, 25～35% alcohol elutions, 95% alcohol elution;
(3) 25～35% alcohol elutions of getting step (2) are through silica gel column chromatography separation, use chloroform-methanol gradient elution, collect the cut E that chloroform-methanol ratio is 9: 1, cut E is separated through ODS column chromatography, use methanol-water gradient elution, collect the cut E-2 that methanol-water ratio is 1: 1, cut E-2 is separated through HW-40 column chromatography, use methanol-water gradient elution, collect the cut E-2-3 that methanol-water ratio is 6: 4, cut E-2-3 is separated through preparation liquid phase HPLC, obtains the compounds of this invention.
3. preparation method according to claim 2, is characterized in that, the described preparative liquid chromatography of step (3), the methanol-water that the ratio of take is 5: 5 is moving phase, detect wavelength and be 254 and 325nm, flow velocity 8mL/min, the retention time in preparation liquid phase is 38.8min.
4. the application of compound in preparation treatment hand foot mouth disease medicine described in claim 1.
5. a pharmaceutical composition, is characterized in that, comprises compound described in claim 1.
6. a pharmaceutical composition that is used for the treatment of hand foot mouth disease, is characterized in that, contains compound and pharmaceutically acceptable carrier described in the claim 1 for the treatment of significant quantity.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310145395.XA CN104098632A (en) | 2013-04-10 | 2013-04-10 | Iridoid glycoside compound, and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310145395.XA CN104098632A (en) | 2013-04-10 | 2013-04-10 | Iridoid glycoside compound, and preparation method and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN104098632A true CN104098632A (en) | 2014-10-15 |
Family
ID=51667208
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310145395.XA Pending CN104098632A (en) | 2013-04-10 | 2013-04-10 | Iridoid glycoside compound, and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104098632A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109053831A (en) * | 2018-09-04 | 2018-12-21 | 山东中医药大学附属医院 | New compound and application in a kind of oldenlandia diffusa |
| CN109503684A (en) * | 2018-12-20 | 2019-03-22 | 广州市香雪制药股份有限公司 | Iridoid glycosides compound and its preparation method and application |
| CN114249783A (en) * | 2021-12-20 | 2022-03-29 | 山东省分析测试中心 | Phenyl succinamide glycoside compound and preparation method and application thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0505572A1 (en) * | 1990-10-09 | 1992-09-30 | TSUMURA & CO. | Iridoide derivative and its use as medicine |
| CN1706859A (en) * | 2005-05-24 | 2005-12-14 | 中国人民解放军第二军医大学 | Process of preparing high-purity jasminodin with Cape jasmine fruit |
| US20070275006A1 (en) * | 2006-03-08 | 2007-11-29 | Council Of Scientific And Industrial Research | Iridoid glycoside composition |
| CN101704857A (en) * | 2009-09-25 | 2010-05-12 | 中国中医科学院中药研究所 | Process for separating and purifying special component specnuezhenide of glossy privet fruit |
-
2013
- 2013-04-10 CN CN201310145395.XA patent/CN104098632A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0505572A1 (en) * | 1990-10-09 | 1992-09-30 | TSUMURA & CO. | Iridoide derivative and its use as medicine |
| CN1706859A (en) * | 2005-05-24 | 2005-12-14 | 中国人民解放军第二军医大学 | Process of preparing high-purity jasminodin with Cape jasmine fruit |
| US20070275006A1 (en) * | 2006-03-08 | 2007-11-29 | Council Of Scientific And Industrial Research | Iridoid glycoside composition |
| CN101704857A (en) * | 2009-09-25 | 2010-05-12 | 中国中医科学院中药研究所 | Process for separating and purifying special component specnuezhenide of glossy privet fruit |
Non-Patent Citations (1)
| Title |
|---|
| HAI-BO LI，等: "Iridoid and bis-iridoid glucosides from the fruit of Gardenia jasminoides", 《FITOTERAPIA》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109053831A (en) * | 2018-09-04 | 2018-12-21 | 山东中医药大学附属医院 | New compound and application in a kind of oldenlandia diffusa |
| CN109503684A (en) * | 2018-12-20 | 2019-03-22 | 广州市香雪制药股份有限公司 | Iridoid glycosides compound and its preparation method and application |
| CN114249783A (en) * | 2021-12-20 | 2022-03-29 | 山东省分析测试中心 | Phenyl succinamide glycoside compound and preparation method and application thereof |
| CN114249783B (en) * | 2021-12-20 | 2023-09-01 | 北京东方红航天生物技术股份有限公司 | Phenyl butanediamide glycoside compound and preparation method and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103254259A (en) | Iridoid glycoside compound as well as preparation method and application thereof | |
| CN105315147A (en) | Guaiane-type sesquiterpenoids compound, and preparation method and application thereof | |
| CN104098632A (en) | Iridoid glycoside compound, and preparation method and application thereof | |
| CN104098630A (en) | Iridoid glycoside compound, and preparation method and application thereof | |
| CN105175383A (en) | Biphenyl compound and preparation method and application thereof | |
| CN105669617A (en) | Lignin compound as well as preparation method and application thereof | |
| CN101538272B (en) | Phenyl propanoid derivative, preparation method thereof and application thereof to preparation of medicines resisting breast cancer | |
| CN113214340A (en) | Antitumor glycyrrhetinic acid derivative and preparation method thereof | |
| CN104072551A (en) | Dipolyiridoid glycoside compound and preparation method and application thereof | |
| CN103254263B (en) | Cardiac glycoside compound and preparation method as well as application thereof | |
| CN104817448B (en) | The application in preparing resisting tobacco mosaic virus medicine of a kind of chalcone compounds | |
| CN105503979B (en) | A kind of compound and its preparation method and application | |
| CN105384746A (en) | Method for extraction of tazettine from plant hymenocallis littoralis bulb | |
| CN105461545A (en) | Cycloreversed cadinane sesquiterpene compound, and preparation method and application thereof | |
| CN104098631A (en) | Iridoid glycoside compound, and preparation method and application thereof | |
| CN103951723A (en) | Golden camellia flavonoid glycoside as well as preparation method and application thereof | |
| CN100434419C (en) | Compound of monocyclic polysubstitution saturated cyclohexanones, prepartion method and usage | |
| CN109053641B (en) | Dineolignan compound, and separation preparation method and application thereof | |
| CN1323078C (en) | Pharmacologically active novel dauer pheromone compound for controlling aging and stress and method for isolating and characterizing the same | |
| CN108347915A (en) | A kind of frozen stock solution and preparation method thereof for adult stem cell long-term preservation | |
| CN110368389A (en) | The preparation method and applications and Claritin of phenolic compound | |
| CN114409557B (en) | Carbon keratin with neuroprotective activity and preparation method and application thereof | |
| TWI764744B (en) | Compound and pharmaceutical composition, use and process extracting from penicillium citrinum thereof | |
| CN113024494B (en) | Phenanthrene compound, preparation method and application | |
| CN107513085B (en) | Compound and preparation method and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| REG | Reference to a national code |
Ref country code: HK Ref legal event code: DE Ref document number: 1197913 Country of ref document: HK |
|
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20141015 |
|
| REG | Reference to a national code |
Ref country code: HK Ref legal event code: WD Ref document number: 1197913 Country of ref document: HK |