CN105418562A - Diterpenoid compound used for treating the prostatic cancer and preparation method therefor - Google Patents
Diterpenoid compound used for treating the prostatic cancer and preparation method therefor Download PDFInfo
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Abstract
The invention discloses a diterpenoid compound used for treating the prostatic cancer and a preparation method therefor, and belongs to the medicine technology field. The provided compound is reported for the first time, is a diterpenoid compound with a novel structure, and can be obtained from dried ternate buttercup roots through extraction, separation and purification. In vitro tests prove that the compound has an inhibition effect on prostatic cancer cell strains PC3 and DU145, and the inhibition effect has a concentration-time dependent relation. The compound (I) can become a potential alternative for late period prostatic cancer treatment, and can be developed into a medicine for treating prostatic cancer.
Description
Technical field
The invention belongs to technical field of pharmaceuticals, be specifically related to from the Root of Catclaw Buttercup of drying, be separated a kind of diterpene-kind compound obtained, containing its pharmaceutical composition and its preparation method and application.
Background technology
Root of Catclaw Buttercup RanunculiTernatiRadix is the dried root of ranunculaceae plant Seynuns Kugelann RanunculusternatusThunb..This product is fusiform, and many 5 ~ 6 cluster, likeness in form ram's horn, and long 3 ~ 10mm, diameter 2 ~ 3mm, there are the residual stem of tawny or stem trace in top.Surface tawny or lark, color and luster stored for a long time deepens, and micro-have vertical wrinkle, and has point-like mark of fibrous root and residual fibrous root.Matter is solid, section off-white color or yellow-white, hollow or solid, mealiness.Gas is micro-, and taste is micro-sweet.Root of Catclaw Buttercup another name three loose grass (Zhejiang), ram's horn grass (Henan), ram's horn (Henan), Root of Whiteflower Hogfennel (Anhui), white melitot (Guangxi).Root of Catclaw Buttercup is warm in nature, and taste is sweet, pungent, enters liver, lung channel, has clearing heat and detoxicating, effect such as dispersing swelling and dissipating binds, cough-relieving apophlegmatic, is clinically used for the treatment of the diseases such as pulmonary tuberculosis, tuberculous lymphadenitis, pharyngolaryngitis, malaria.Among the peoplely treat lymphoid tuberculosis with Root of Catclaw Buttercup, no matter tuberculosis size or whether suppurate and all have good curative effect.Study hotspot is become in recent years because it has good antitumous effect.
Research in recent years for Root of Catclaw Buttercup chemical composition is less, and it is mainly containing the compound such as flavonoid and glycoside, alkaloid, volatile oil, organic acid.
It is reported, Root of Catclaw Buttercup decoction treatment lymphoid tuberculosis 180 example, it is efficient reaches 100%, clinical cure rate 73.9%; Its injection liquid has its decoction of restraining effect all to have remarkable restraining effect to dysentery bacterium, streptococcus aureus, Staphylococcus albus, tetrads etc. to the strain of mouse S180, S37 cancer.Separately there is report Root of Catclaw Buttercup energy induced tumors necrosin (TNF).Tumour necrosis factor is one group and has antitumor, antiviral, to participate in panimmunity regulate process sphaeroprotein, and because it can kill tumour cell specifically, and normal tissue cell has no adverse effects, thus plays the effect of tumour immunity and oncotherapy.
Summary of the invention
The object of this invention is to provide and a kind ofly from the Root of Catclaw Buttercup of drying, be separated a kind of diterpene-kind compound obtained, containing its pharmaceutical composition and its preparation method and application.
Above-mentioned purpose of the present invention is achieved by technical scheme below:
There is the compound (I) of following structural formula:
The preparation method of described compound (I), comprise following operation steps: the Root of Catclaw Buttercup of drying is pulverized by (a), extract with 70 ~ 80% alcohol heat reflux, united extraction liquid, be concentrated into without alcohol taste, use sherwood oil, ethyl acetate and water saturated n-butanol extraction successively, obtain petroleum ether extract, acetic acid ethyl ester extract and n-butyl alcohol extract respectively; Acetic acid ethyl ester extract macroporous resin removal of impurities in (b) step (a), first use 10% ethanol elution, 6 column volumes, then use 70% ethanol elution, 8 column volumes, collect 70% elutriant, concentrating under reduced pressure obtains 70% ethanol elution enriched material; C in () step (b), 70% ethanol elution enriched material purification on normal-phase silica gel is separated, obtain 5 components successively with the methylene chloride-methanol gradient elution that volume ratio is 85:1,50:1,25:1,10:1 and 1:1; D in () step (c), component 4 is separated further by purification on normal-phase silica gel, obtain 3 components successively with the methylene chloride-methanol gradient elution that volume ratio is 15:1,10:1 and 5:1; E in () step (d), component 2 reverse phase silica gel of octadecylsilane bonding is separated, with the methanol aqueous solution isocratic elution that concentration expressed in percentage by volume is 70%, collect 8 ~ 12 column volume elutriants, elutriant concentrating under reduced pressure obtains pure compound (I).
Further, in step (a), extract with 75% alcohol heat reflux, united extraction liquid.
Further, described macroporous resin is D101 type macroporous adsorbent resin.
A kind of pharmaceutical composition, the described compound (I) wherein containing treatment significant quantity and pharmaceutically acceptable carrier.
The application of described compound (I) in the medicine of preparation treatment prostate cancer.
The application of described pharmaceutical composition in the medicine of preparation treatment prostate cancer.
When the compounds of this invention is used as medicine, directly can uses, or use with the form of pharmaceutical composition.
This pharmaceutical composition contains the compounds of this invention (I) for the treatment of significant quantity, and all the other are acceptable on pharmacology, nontoxic to humans and animals and pharmaceutically acceptable carrier of inertia and/or vehicle.
Described pharmaceutically acceptable carrier or vehicle are that one or more are selected from solid, semisolid and liquid diluent, filler and pharmaceutical preparation assistant agent.Pharmaceutical composition of the present invention is used with the form of per weight dose.Medicine of the present invention is applied to by form that is oral or injection the patient needing treatment.For time oral, tablet, slow releasing tablet, controlled release tablet, capsule, dripping pill, micropill, suspensoid, emulsion, powder or granule, oral liquid etc. can be made into; During for injecting, can be made into water-based or oily solution, aseptic powder injection, liposome or the emulsion etc. of sterilizing.
Accompanying drawing explanation
Fig. 1 is compound (I) structural formula;
Fig. 2 is that compound (I) calculates ECD and experiment ECD figure;
Fig. 3 is PC3 and DU145 survival rate after different concns compound (I) effect 72h;
Fig. 4 is PC3 and DU145 survival rate after 10.0mg/L compound (I) effect different time.
Embodiment
Further illustrate essentiality content of the present invention below in conjunction with embodiment, but do not limit scope with this.Although be explained in detail the present invention with reference to preferred embodiment, those of ordinary skill in the art should be appreciated that and can modify to technical scheme of the present invention or equivalent replacement, and does not depart from essence and the scope of technical solution of the present invention.
Main raw, reagent source and instrument type:
Ethanol, sherwood oil, ethyl acetate, propyl carbinol, methylene dichloride are analytical pure, and purchased from Shanghai Ling Feng chemical reagent company limited, methyl alcohol, analytical pure, purchased from Jiangsu Han Bang chemical reagent company limited.
Embodiment 1: compound (I) is separated preparation and structural identification
A Root of Catclaw Buttercup (10kg) pulverizing that () is dry, (30L × 3 time) are extracted with 75% alcohol heat reflux, united extraction liquid, be concentrated into without alcohol taste (6L), use sherwood oil (6L × 3 time), ethyl acetate (6L × 3 time) and water saturated propyl carbinol (6L × 3 time) to extract successively, obtain petroleum ether extract, acetic acid ethyl ester extract (381g) and n-butyl alcohol extract respectively; Acetic acid ethyl ester extract D101 macroporous resin removal of impurities in (b) step (a), first use 10% ethanol elution, 6 column volumes, use 70% ethanol elution, 8 column volumes again, collect 70% elutriant, concentrating under reduced pressure obtains 70% ethanol elution enriched material (131g); C in () step (b), 70% ethanol elution enriched material purification on normal-phase silica gel is separated, successively with volume ratio be 85:1 (8 column volumes), the methylene chloride-methanol gradient elution of 50:1 (8 column volumes), 25:1 (8 column volumes), 10:1 (10 column volumes) and 1:1 (8 column volumes) obtains 5 components; D component 4 (27g) is separated further by purification on normal-phase silica gel in () step (c), successively with volume ratio be 15:1 (8 column volumes), the methylene chloride-methanol gradient elution of 10:1 (8 column volumes) and 5:1 (6 column volumes) obtains 3 components; E in () step (d), component 2 (18g) reverse phase silica gel of octadecylsilane bonding is separated, with the methanol aqueous solution isocratic elution that concentration expressed in percentage by volume is 70%, collect 8 ~ 12 column volume elutriants, elutriant concentrating under reduced pressure obtains pure compound (I) (230mg).
Structural identification: white amorphous powder; HR-ESIMS shows [M+Na]
+for m/z513.1702, can obtain molecular formula in conjunction with nuclear-magnetism feature is C
25h
30o
10, degree of unsaturation is 11.Hydrogen nuclear magnetic resonance modal data δ
h(ppm, DMSO-d
6, 600MHz): H-1 (5.19, s), H-3 (2.41, m), and H-3 (2.16, m), H-6 (5.25, s), and H-8 (2.82, m), H-9 (2.61, t, J=12.0), and H-11 (1.35, m), H-11 (1.99, m), H-15 (5.93, s), H-17 (5.54, d, J=1.8), H-17 (5.32, d, J=1.8), and H-18 (1.01, s), H-19 (1.01, s), H-20 (1.11, s), and 1-OAc (2.04, s), 6-OAc (2.14, s), 12-OCH
3(3.02, s), 5-OH (2.89, br, s); Carbon-13 nmr spectra data δ
c(ppm, DMSO-d
6, 125MHz): 81.7 (CH, 1-C), 201.7 (C, 2-C), 47.7 (CH
2, 3-C), 38.3 (C, 4-C), 91.2 (C, 5-C), 90.4 (CH, 6-C), 202.4 (C, 7-C), 48.1 (CH, 8-C), 36.5 (CH, 9-C), 43.8 (C, 10-C), 35.4 (CH
2, 11-C), 116.6 (C, 12-C), 166.0 (C, 13-C), 139.4 (C, 14-C), 115.1 (CH, 15-C), 169.3 (C, 16-C), 115.7 (CH
2, 17-C), 30.2 (CH
3, 18-C), 24.3 (CH
3, 19-C), 16.4 (CH
3, 20-C), 168.8 (C, 1-OAc), 20.7 (CH
3, 1-OAc), 172.1 (C, 6-OAc), 21.7 (CH
3, 6-OAc), 49.6 (CH
3, 12-OCH
3); Carbon atom mark is see Fig. 1.
13cNMR spectrum and DEPT spectrum show 25 carbon signals, comprising six methyl (methoxyl group), three methylene radical (an olefinic methylene base), five methyne (olefinic methynes, two containing oxygen methyne), and 11 quaternary carbons (two containing oxygen quaternary carbon for two alkene carbon, five carbonyl carbon).IR stave this compound bright contains hydroxyl and carbonyl (3510 and 1740cm
-1) group, it has maximal ultraviolet absorption at 218nm place in addition, shows that this compound contains α, β-unsaturated butenolide structure.Olefinic proton signals δ H5.93 (H-15, s) and be positioned at the carbon signal δ C166.0 (C-13) of low field, 115.1 (C-15) and 169.3 (C-16) also demonstrate that the existence of this structure.In addition in conjunction with three methyl signals δ H1.01 (H
3-18, s), 1.01 (H
3-19, s) He 1.11 (H
3-20, s), show that this compound is a tetracyclic diterpene compound containing butenolide structural unit.In HMBC spectrum, H-1 (δ H5.19, s) and-OCOCH
3(δ C168.8); H-6 (δ H5.25, s) and-OCOCH
3(δ C172.1); 12-OCH
3with the dependency of C-12 (δ C116.6), (δ H3.02, s) shows that C-1 and C-6 position is connected with acetoxyl group, C-12 is connected with methoxyl group.In HSQC and HMBC spectrum, proton signal δ H5.54 (d, J=1.8Hz) and 5.32 (d, J=1.8Hz) with carbon signal (δ C115.7, C-17) be directly connected, and with the dependency of C-14 (δ C139.4) and C-13 (δ C166.0) long distance, show to be connected with double bond between C-14 and C-17.Comprehensive hydrogen spectrum, carbon spectrum, HMBC spectrum and NOESY spectrum, and document is about correlation type nuclear magnetic data, can substantially determine this compound as shown in Figure 1, steric configuration is determined further by ECD test, theoretical value and experimental value basically identical (Fig. 2).
Embodiment 2: compound (I) pharmacological action is tested
One, material and instrument
Prostatic cancer cell lines PC3 (ArCC-CRL-1435), Prostatic cancer cell lines DU145 (ATCC-HTB-81).Compound (I) is made by oneself, and HPLC normalization method purity is greater than 98%, and being mixed with concentration with dimethyl alum (DMSO) is that the storage liquid of 1.0g/L is for subsequent use.CCK-8 test kit is Japanese colleague's chemistry institute product.RPMI-1640 substratum is purchased from Gibco company.Foetal calf serum (FBS) is purchased from Hyelone company.Green grass or young crops/Streptomycin sulphate is Shanghai pioneer's medicine company product.Trypsinase is purchased from Huamei Bio-Engrg Co..Dimethyl sulfoxide (DMSO) (DMSO) is purchased from Shanghai Hua Shun bio-engineering corporation.Agar (Agarose), dithiothreitol (DTT) (DTT), phenmethyl sulfonephthalein fluorine (PMSF), Tetramethyl Ethylene Diamine (TEMED) are Sigma Products.Sodium laurylsulfonate (SDs), trichloromethyl alkyl methane (Tris) Tris-Hcl, Tritonx-100 are Promega Products.Propylene phthalein amine, persulfuric acid money (AP) are purchased from Suzhou chemical reagent factory product.
CO
2cell culture incubator (ShellLab), inverted phase contrast microscope (Nikon), Bechtop (Suzhou Decontamination Equipment Plant), flow cytometer (BD), F039300A type microplate reader (Sunrise), autoclave sterilizer (HirayamaHA-300MD), low temperature ultracentrifuge (Kubota3740), Universal table (Jiangsu kylin medical apparatus factory TS-92), electrophoresis apparatus (Gibco company), LXJ-II type whizzer (Shanghai medical analytical instrument factory), DK600 type electric heating constant-temperature water-bath tank (Shanghai microtest equipment company), electronic balance (METTLERTOLEDO), desk-top loft drier (Shanghai gloomy reliable test Instrument Ltd.), UV detector (Beckman).
Two, test method
1, cell cultures and maintenance
1.1 cell cultures
Cell strain is incubated at centralab of tumour hospital.Be incubated in the RPMI-1640 substratum containing 10% foetal calf serum, another add paddy ammonia phthalein amine (2mmol/L) and microbiotic (l00U/ penicillin and 100mg/L Streptomycin sulphate), put 37 DEG C, saturated humidity, containing 5%CO
2cultivate in the incubator of gas.
1.2 passage
1. under inverted phase contrast microscope, observation of cell covers with adherent, can go down to posterity.Before going down to posterity by 75% alcohol wipe through the Bechtop of uviolizing and both hands.2. suck the old nutrient solution in culturing bottle with suction pipe, clean 3 times with PBS.3. add 0.02%EDTA-0.25% trypsin solution 2mL to digest, leave standstill about 5 minutes, and frequently observe under inverted phase contrast microscope, when kytoplasm retraction, when no longer connecting in blocks between cell, add the appropriate fresh culture containing serum and stop tryptic effect.4. with dropper, the cell digested is blown and beaten into cell suspension, equilibrium centrifugation (1000 revs/min) 5 minutes in suction 10mL centrifuge tube.5. abandoning supernatant, adds 2mL nutrient solution, and blow and beat cell gently with dropper and make cell suspension, 1:2 ~ 3 are distributed into new culturing bottle, adds nutrient solution and continues in right amount to cultivate in incubator.
2, morphological observation
Inverted microscope is observed: logarithmic phase PC3 and DU145 cell are inoculated in 6cm culture dish, adds 10.0mgL after cultivating 24h
-1, observation of cell morphologic change record under inverted phase contrast microscope respectively after compound (I) process 24,48,72h.
3, cytotoxicity test (CCK-8 method)
1. the cell dissociation in culturing bottle becomes single cell suspension, according to 3 × 10 after cell counting
4individual cells/well is inoculated in 96 orifice plates, and every hole adds 0.lmL substratum, cellar culture in 37 DEG C, containing 5%CO
2in the incubator of gas.2. grouping is tested: establish negative control group (to have cell but not dosing, 0.1%DMSO), blank group (acellular only have nutrient solution), compound (I) is 2.5mg/L respectively, 5.0mg/L, 10.0mg/L and 20.0mg/L totally 6 groups.3. after 24 hours, observation of cell adherent growth is good, and divide into groups to dosing in 96 orifice plates by above-mentioned test respectively, often group establishes 6-8 repeating hole.4. after dosing, 96 orifice plates are moved into 37 DEG C, containing 5%CO
2continue in the incubator of gas to cultivate 24,48 and 72 hours respectively.When 5. often organizing off-test, every hole adds CCK-810 μ L, continue in 37 DEG C of incubators cultivation after 4 hours microplate reader detect absorbancy (OD) value in 450 every holes, mensuration wavelength is 450nm, and reference wavelength is 600nm.6. go out cell survival rate (cellviability) according to following formulae discovery, be then depicted as chart, value when survival rate is 50% is IC
50.Cell survival rate (%)=[(As-Ab)/(Ac-Ab)] × 100%.Wherein As is test holes, and Ac is control wells, and Ab is blank well.
4, colony-forming test
1. to take the logarithm cell in vegetative period, with 3 × l0 after counting
2individually be inoculated in 6 orifice plates, every hole adds 2mL substratum, cellar culture in 37 DEG C, containing 5%CO
2in the incubator of gas.2. after 24h, observation of cell adherent growth is good, and add different concns (0,2.5,5.0,10.0,20.0mg/L) compound (I) process respectively, often group establishes 3 repeating holes.3. after dosing, 6 orifice plates are moved into 37 DEG C, containing 5%CO
2continue 14 days in the incubator of gas.4. 10% methyl alcohol is fixed, and Giemsa dyes, and calculates every hole colony number (the counting a colony of >=50 cells).5. colony forming efficiency is calculated: colony sum/inoculating cell number × 100%.
Three, result and conclusion
1, compound (I) gas is on the impact of PC3 and DU145 cellular form
See cellular control unit adherent growth under mirror, flanking cell merges in flakes, and cell is similar round or fusiformis, and volume is comparatively large, and arrangement is tight, the smooth of the edge, and kytoplasm is full, and the structure outline such as nuclear membrane, kernel is obvious, and Growth of Cells is rapid; After 10.0mg/L compound (I) process, cell density reduces gradually, and the speed of growth obviously slows to and almost stagnates, and cell comes off gradually and floats in nutrient solution.Cell volume reduces, after birth shrinkage, becomes small circular or irregular form, common fine granularity material in born of the same parents.Drug treating time is longer, and morphological changes of cell is more obvious.
2, cytotoxicity test
Different concns compound (I) growth to Prostatic cancer cell lines PC3 and DU145 all has restraining effect.2.5,5.0,10.0,20.0mg/L compound (I) acts on the survival rate after two kinds of cells 24,48 and 72h (table l and table 2) as shown in the table.Wherein, the cell survival rate after 10.0mg/L compound (I) acts on PC3 and DU145 cell 24,45,72h is respectively 56.2%, 42.5%, 24.3% and 50.8%, 32.6%, 20.7%; 2.5,5.0,10.0,20.0mg/L, the survival rate after compound (I) effect two kinds of cell 72h is respectively 54.3%, 37.7%, 24.3%, 13.2% and 52.4%, 32.8%, 20.7%, 11.2% (see Fig. 3 and Fig. 4).Two analysis of variance show that between different concns and different time treatment group, difference has statistical significance (P<0.05), and prompting compound (I) the growth-inhibiting effect to PC3 and DU145 is that time-concentration relies on.
3, colony-forming test
Test display, the colony forming efficiency of control group PC3 and DU145 is respectively 67.7 and 64%, and 2.5,5.0,10.0,20.0mg/L compound (I) treatment group is respectively 60.7%, 51.3,39.3%, 27.0% and 49.7%, 34.0%, 27.7%, 15.7% (see table 3).This further demonstrates that compound (I) has inhibited proliferation to PC3 and DU145 cell.
Conclusion, verify that compound (I) is to the effect of Prostatic cancer cell lines PC3 and DU145, and restraining effect has concentration in this test by CCK-8 method and colony-forming test---time-dependent sexual intercourse.Compound (I) may become a potential selection of tool in therapy approaches of advanced prostate cancer.
Table 1 different concns compound (I) acts on the cell survival rate after PC3
| Time/concentration | 2.5mg/L | 5.0mg/L | 10.0mg/L | 20.0mg/L |
| 24h | 0.825 | 0.731 | 0.562 | 0.422 |
| 48h | 0.69 | 0.53 | 0.425 | 0.226 |
| 72h | 0.543 | 0.377 | 0.243 | 0.132 |
Table 2 different concns compound (I) acts on the cell survival rate after DU145
| Time/concentration | 2.5mg/L | 5.0mg/L | 10.0mg/L | 20.0mg/L |
| 24h | 0.809 | 0.604 | 0.508 | 0.375 |
| 48h | 0.646 | 0.481 | 0.326 | 0.173 5 --> |
| 72h | 0.542 | 0.328 | 0.207 | 0.112 |
The colony forming efficiency of lower PC3 and DU145 of table 3 different concns compound (I) effect
Embodiment 3
The preparation of tablet: by embodiment 1 method first obtained compound (I), and the salt utilizing organic acid to make as tartrate or citric acid or formic acid or oxalic acid etc., mineral acid example hydrochloric acid or sulfuric acid or phosphoric acid, vehicle is added, pelletizing press sheet than the ratio for 1:7 in itself and excipient weight.
Embodiment 4
The preparation of oral liquid: by embodiment 1 method first obtained compound (I), and the salt utilizing organic acid to make as tartrate or citric acid or formic acid or oxalic acid etc., mineral acid example hydrochloric acid or sulfuric acid or phosphoric acid, oral liquid method for making makes oral liquid routinely.
Embodiment 5
The preparation of capsule or granule: by embodiment 1 method first obtained compound (I), and the salt utilizing organic acid to make as tartrate or citric acid or formic acid or oxalic acid etc., mineral acid example hydrochloric acid or sulfuric acid or phosphoric acid, add vehicle in itself and excipient weight than the ratio for 1:9, make capsule or granule.
Embodiment 6
The preparation of injection liquid: by embodiment 1 method first obtained compound (I), and the salt utilizing organic acid to make as tartrate or citric acid or formic acid or oxalic acid etc., mineral acid example hydrochloric acid or sulfuric acid or phosphoric acid, inject with water routinely, essence filter, injection liquid is made in embedding sterilizing.
Embodiment 7
The preparation of aseptic powder injection: by embodiment 1 method first obtained compound (I), and the salt utilizing organic acid to make as tartrate or citric acid or formic acid or oxalic acid etc., mineral acid example hydrochloric acid or sulfuric acid or phosphoric acid, be dissolved in sterile water for injection, stirring makes molten, filter with aseptic suction funnel, aseptic essence filter again, be sub-packed in ampoule, after frozen drying, aseptic sealing by fusing obtains powder injection.
The effect of above-described embodiment is essentiality content of the present invention is described, but does not limit protection scope of the present invention with this.Those of ordinary skill in the art should be appreciated that and can modify to technical scheme of the present invention or equivalent replacement, and does not depart from essence and the protection domain of technical solution of the present invention.
Claims (7)
1. there is the compound (I) of following structural formula:
2. the preparation method of compound according to claim 1 (I), it is characterized in that comprising following operation steps: the Root of Catclaw Buttercup of drying is pulverized by (a), extract with 70 ~ 80% alcohol heat reflux, united extraction liquid, be concentrated into without alcohol taste, use sherwood oil, ethyl acetate and water saturated n-butanol extraction successively, obtain petroleum ether extract, acetic acid ethyl ester extract and n-butyl alcohol extract respectively; B in () step (a), the removal of impurities of thing macroporous resin got by propyl carbinol, first use 10% ethanol elution, 6 column volumes, then use 70% ethanol elution, 8 column volumes, collect 70% elutriant, concentrating under reduced pressure obtains 70% ethanol elution enriched material; C in () step (b), 70% ethanol elution enriched material purification on normal-phase silica gel is separated, obtain 5 components successively with the methylene chloride-methanol gradient elution that volume ratio is 85:1,50:1,25:1,10:1 and 1:1; D in () step (c), component 4 is separated further by purification on normal-phase silica gel, obtain 3 components successively with the methylene chloride-methanol gradient elution that volume ratio is 15:1,10:1 and 5:1; E in () step (d), component 2 reverse phase silica gel of octadecylsilane bonding is separated, with the methanol aqueous solution isocratic elution that concentration expressed in percentage by volume is 70%, collect 8 ~ 12 column volume elutriants, elutriant concentrating under reduced pressure obtains pure compound (I).
3. the preparation method of compound according to claim 2 (I), is characterized in that: in step (a), extracts, united extraction liquid with 75% alcohol heat reflux.
4. the preparation method of compound according to claim 2 (I), is characterized in that: described macroporous resin is D101 type macroporous adsorbent resin.
5. a pharmaceutical composition, is characterized in that: the compound according to claim 1 (I) wherein containing treatment significant quantity and pharmaceutically acceptable carrier.
6. the application of compound according to claim 1 (I) in the medicine of preparation treatment prostate cancer.
7. the application of pharmaceutical composition according to claim 5 in the medicine of preparation treatment prostate cancer.
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| CN201511016208.3A CN105418562A (en) | 2015-12-29 | 2015-12-29 | Diterpenoid compound used for treating the prostatic cancer and preparation method therefor |
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| CN201511016208.3A Withdrawn CN105418562A (en) | 2015-12-29 | 2015-12-29 | Diterpenoid compound used for treating the prostatic cancer and preparation method therefor |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105330714A (en) * | 2015-11-25 | 2016-02-17 | 杨飞杰 | Novel limonin compound as well as preparation method and medical application thereof in treatment of prostatic cancer |
| CN105330716A (en) * | 2015-10-21 | 2016-02-17 | 淄博夸克医药技术有限公司 | New withanolides compound, and preparation method and medical application thereof |
| CN105503800A (en) * | 2016-02-20 | 2016-04-20 | 杭州富阳伟文环保科技有限公司 | Novel eremophilanolides type compound and preparation method and medical application thereof |
| CN105503782A (en) * | 2016-01-04 | 2016-04-20 | 范素琴 | Protoilludanes type sesquiterpenoid compound and preparation method and medical application thereof |
| CN106083974A (en) * | 2016-06-03 | 2016-11-09 | 郑巧丹 | A kind of new lanostane-type triterpene compounds and preparation method thereof and medical usage |
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2015
- 2015-12-29 CN CN201511016208.3A patent/CN105418562A/en not_active Withdrawn
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105330716A (en) * | 2015-10-21 | 2016-02-17 | 淄博夸克医药技术有限公司 | New withanolides compound, and preparation method and medical application thereof |
| CN105330714A (en) * | 2015-11-25 | 2016-02-17 | 杨飞杰 | Novel limonin compound as well as preparation method and medical application thereof in treatment of prostatic cancer |
| CN105503782A (en) * | 2016-01-04 | 2016-04-20 | 范素琴 | Protoilludanes type sesquiterpenoid compound and preparation method and medical application thereof |
| CN105503800A (en) * | 2016-02-20 | 2016-04-20 | 杭州富阳伟文环保科技有限公司 | Novel eremophilanolides type compound and preparation method and medical application thereof |
| CN106083974A (en) * | 2016-06-03 | 2016-11-09 | 郑巧丹 | A kind of new lanostane-type triterpene compounds and preparation method thereof and medical usage |
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Application publication date: 20160323 |