CN105061456A - Novel kaurene diterpenoid compound and preparation method and pharmaceutical application thereof - Google Patents
Novel kaurene diterpenoid compound and preparation method and pharmaceutical application thereof Download PDFInfo
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Abstract
The invention discloses a novel kaurene diterpenoid compound and a preparation method and pharmaceutical application thereof, belongs to the field of medicines, and particularly relates to the novel kaurene diterpenoid compound separated from the dried leaves of sweet viburnum, the preparation method and the pharmaceutical application of the compound. The compound, which is reported for the first time, can be obtained by extraction, separation and purification from the dried leaves of the sweet viburnum, and the purity is high. An experiment proves that the novel aurane diterpenoid compound can remarkably inhibit the growth of melanoma cells, moreover, the number of the cells and the concentration of a compound (I) decrease dependently, and therefore medicines for treating melanomas can be researched and developed from the novel kaurene diterpenoid compound.
Description
Technical field
The invention belongs to pharmaceutical field, be specifically related to be separated a kind of new kauri pine alkanes diterpene compound obtained and preparation method thereof and medicinal use from the dry leave of sweet viburnum.
Background technology
Sweet viburnum is Caprifoliaceae (Caprifoliaceae) Viburnum plant (Viburnumodoratissimum), evergreen shrubs or dungarunga, another name: high smoke tree tree, root of Sweet Viburnum, coral branch, france mongolicus.Originate in China, be mainly distributed in Hunan, Fujian, Guangdong, Taiwan and Zhejiang, also there are distribution in Japan, Korea.The former mutation of sweet viburnum is ViburnumodoratissimumKer-Gawl.var.odoratissimum, and other two mutation are respectively Japanese sweet viburnum (ViburnumodoratissimumKer-Gawl.var.awabuki) and Yunnan sweet viburnum (ViburnumodoratissimumKer-Gawl.var.sessiliflorum); Begin to be loaded in " Chinese Higher plant illustrated handbook ", its branches and leaves can be used as medicine, and have clearing and activating the channels and collaterals effect, cure mainly rheumatic arthralgia, treating swelling and pain by traumatic injury, fracture.
Main containing compounds such as diterpene, triterpene, flavones, sesquiterpene, lignanoid and coumarin glycosides in sweet viburnum.Wherein vibsanin type diterpene-kind compound is the characteristic compounds of sweet viburnum, according to modern study report, it is only present in V.awabuki, V.odoratissimum and V.suspensum of Viburnum, it is divided into that 3 kinds of hypotype structures i.e. ten unitary are ring-like, seven-membered ring type and rearrangement type, and these 3 kinds of subtype compounds rearrangement reaction can occur under high light or high temperature.
Sweet viburnum mainly has cell toxicant, antibacterial, ichthyotoxin, suppression plant-growth isoreactivity, wherein cytotoxic activity is its primary bioactivity, and Recent study finds that in sweet viburnum, composition goes out cytotoxic activity to cells show such as KB, A-549, HT-29, P-388, NUGC-3 and HONE-1.In addition, the organic acid separated from sweet viburnum and flavone component have stronger fungistatic effect.
Summary of the invention
The object of the present invention is to provide and a kind ofly from the dry leave of sweet viburnum, be separated a kind of new kauri pine alkanes diterpene compound obtained;
Another object of the present invention is to the preparation method that this new compound is provided;
Another object of the present invention is the medicinal use providing this compound for the preparation for the treatment of melanoma medicine.
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 dry rhizome of (a) sweet viburnum is pulverized, extract with 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), use 8% ethanol and 75% ethanol elution successively, collect 75% ethanol eluate, concentrating under reduced pressure obtains 75% ethanol elution thing medicinal extract; C in () step (b), 75% ethanol elution medicinal extract purification on normal-phase silica gel is separated, obtain 5 components successively with the methylene chloride-methanol gradient elution that volume ratio is 80:1,45:1,20:1,10:1 and 1:1; D in () step (c), component 3 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 25:1,20:1 and 15:1; 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 elutriant, elutriant concentrating under reduced pressure obtains pure compound (I).
Further, described macroporous resin is AB-8 type macroporous adsorbent resin.
Pharmaceutical composition, the described compound (I) wherein containing treatment significant quantity and pharmaceutically acceptable carrier.
The application of described compound (I) in the melanomatous medicine of preparation treatment.
The application of described pharmaceutical composition in the melanomatous medicine of preparation treatment.
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.
Figure of description
Fig. 1 is compound (I) structural formula;
Fig. 2 is that the theoretical ECD value of compound (I) compares with experiment ECD value;
Fig. 3 is BrdU Immunofluorescence test Cell proliferation results.
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.
Embodiment 1: compound (I) is separated preparation and structural identification
Medicinal material and reagent source: ethanol, sherwood oil, ethyl acetate, propyl carbinol, methylene dichloride are analytical pure, purchased from Shanghai Ling Feng chemical reagent company limited, methyl alcohol, analytical pure, purchased from Jiangsu Han Bang chemical reagent company limited.Sweet viburnum medicinal material purchased from Hui nationality's Chinese Medicinal Materials Markets, Fujian, the place of production.
Preparation method: the dry leave (8kg) of sweet viburnum is pulverized by (a), (25L × 3 time) are extracted with 80% alcohol heat reflux, united extraction liquid, be concentrated into without alcohol taste (6L), sherwood oil (6L × 3 time), ethyl acetate (6L × 3 time) and water saturated propyl carbinol (6L × 3 time) is used to extract successively, concentrated, obtain petroleum ether extract, acetic acid ethyl ester extract (315g) and n-butyl alcohol extract respectively; B in () step (a), acetic acid ethyl ester extract dissolved in purified water is to 2L, medical absorbent cotton filters, filtrate is separated with AB-8 type macroporous resin (1.5kg), use 8% ethanol (10L) and 75% (12L) ethanol elution successively, collect 75% ethanol eluate, concentrating under reduced pressure obtains 75% ethanol elution thing medicinal extract (155g); C in () step (b), 75% ethanol elution medicinal extract 200-300 order purification on normal-phase silica gel is separated, successively with volume ratio be 80:1 (10 column volumes), the methylene chloride-methanol gradient elution of 45:1 (8 column volumes), 20:1 (8 column volumes), 10:1 (8 column volumes) and 1:1 (5 column volumes) obtains 5 components; D component 3 (36g) 200-300 order purification on normal-phase silica gel is separated further in () step (c), successively with volume ratio be 25:1 (8 column volumes), the methylene chloride-methanol gradient elution of 20:1 (8 column volumes) and 15:1 (5 column volumes) obtains 3 components; E in () step (d), component 2 (12g) the reverse phase silica gel ODS-C18 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 elutriant, elutriant concentrating under reduced pressure obtains pure compound (I) (25mg).
Structural identification: HR-ESIMS shows [M+Na]
+for m/z401.1614, can obtain molecular formula in conjunction with nuclear-magnetism feature is C
20h
26o
7, degree of unsaturation is 8.Hydrogen nuclear magnetic resonance modal data δ
h(ppm, DMSO-d
6, 500MHz): H-1 (5.98, dd, J=11.9, 5.8), H-2 (2.36, m), H-2 (2.29, overlap), H-3 (3.90, m), H-5 (2.84, br, s), H-6 (5.95, br, s), H-9 (2.89, d, J=3.6), H-11 (5.07, m), H-12 (2.27, overlap), H-12 (1.99, m), H-13 (2.64, m), H-14 (3.69, d, J=10.8), H-14 (3.05, dd, J=10.8, 4.0), H-17 (1.43, s), H-18 (1.27, s), H-19 (1.22, s), H-20 (4.55, d, J=8.8), H-20 (4.57, d, J=8.8), 3-OH (4.77, s), 16-OH (4.62, s), carbon-13 nmr spectra data δ
c(ppm, DMSO-d
6, 125Hz): 76.7 (CH, 1-C), 31.9 (CH
2, 2-C), 72.4 (CH, 3-C), 37.3 (C, 4-C), 54.9 (CH, 5-C), 101.5 (CH, 6-C), 171.4 (C, 7-C), 56.4 (C, 8-C), 49.8 (CH, 9-C), 48.9 (C, 10-C), 64.7 (CH, 11-C), 33.5 (CH
2, 12-C), 40.5 (CH, 13-C), 32.6 (CH
2, 14-C), 211.8 (C, 15-C), 77.7 (C, 16-C), 19.9 (CH
3, 17-C), 29.1 (CH
3, 18-C), 15.8 (CH
3, 19-C), 73.4 (CH
2, 20-C), carbon atom mark is see Fig. 1.
13this compound of CNMR and DEPT spectrum display contains three methyl, four methylene radical (containing oxygen), seven methynes (four containing oxygen), six quaternary carbons (contains oxygen carbon, a ketone carbonyl carbon and a lactone carbonyl carbon).H in HMBC spectrum
2-14 (δ H3.69 and 3.05), H-13 (δ H2.64), H
2-12 (δ H2.27 and 1.99) and Me-17 (δ H1.43) show C-16 position to be connected with an oh group with the dependency containing oxygen quaternary carbon C-16 (δ C77.7).H in HMBC spectrum
2the dependency of-2 (δ H2.36 and 2.29), Me-18 (δ H1.27) and Me-19 (δ H1.22) and oxidation methyne C-3 (δ C72.4), and
1h-
1in HCOSY spectrum, the dependency of H-2 and H-3, shows existence oh group on C-3 position.H-1 and H-3, H-5 β and Me-18 in ROESY spectrum, and the dependency of Me-17 and H-12 α shows that C-3-OH and C-16-OH is respectively α and beta comfiguration.Comprehensive hydrogen spectrum, carbon spectrum, HMBC spectrum and ROESY 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
People A375 melanoma cell strain is given by Third Military Medical University.Compound (I) is made by oneself, and preparation method is shown in that embodiment 1, HPLC normalization method purity is greater than 98%.DMEM substratum, 0.25% pancreatin purchased from American Gibco company.MTT (3-(4,5-dimethylthiazole-2)-2,5-diphenyltetrazolium bromide bromine salt; Methyl thiazoly tetrazolium assay), DMSO (dimethyl sulfoxide (DMSO)), BrdU purchased from American Sigma company.Mouse-anti BrdU polyclonal antibody purchased from American ABcam company.Goat against murine two anti-purchased from American Cellsignaling company.AnnexinV-FITC/PI cell apoptosis detection kit purchased from American Promega company.
Constant temperature CO
2incubator, general refrigerator ,-80 degree refrigerators are U.S. Forma Products.Bechtop (Chinese Suzhou cleaning project company).Inverted microscope, inverted fluorescence microscope (olympus company).Electro-heating standing-temperature cultivator (Chinese Shanghai leap medical apparatus and instruments factory).Automatic microplate reader (Japanese Wako company).UV detector (Beckman company of the U.S.).J6-HC supercentrifuge (Beckman company of the U.S.).Low-temperature trace whizzer (German Eppendorf company).Vibration shaking table (Forma company of the U.S.).
Two, test method
1, cell cultures
1.1 cell recovery
Frozen A375 melanoma cell in liquid nitrogen container taken out, put into rapidly the warm water of 37 DEG C, shake makes it melt as early as possible (about lmin) gently.Then sucking-off cell suspension, joins in the centrifuge tube being added with 2mL substratum, blows and beats mixing gently, 800r/min, and centrifugal 5min discards upper strata substratum.After making cell suspension with the DMEM substratum 8mL of the mould G/ Streptomycin sulphate containing 10%FBS and 1%, be seeded in 10cm culture plate.Then 5%CO is placed in
2, cultivate in 37 DEG C of constant incubators.
1.2 passage
Basis of microscopic observation cytogamy degree can go down to posterity when reaching 80%-90%.First use 2mLPBS washed cell 2 times, then add the pancreatin lmL of 0.25%.Horizontal wave and culture dish gently, makes Digestive system can cover the surface of cell, is then placed in 37 DEG C of incubators and digests about lmin.See that cell rounding bounces back under being placed in microscope, then add rapidly the substratum termination digestion that 1mL contains FBS.Blow and beat to cell dispersal even gently, then go down to posterity according to 1:2 or 1:3 according to cell density, be re-seeded in new culture plate, be placed in 5%CO
2, cultivate in 37 DEG C of constant incubators.
1.3 cell cryopreservation
Get the cell that 1 dish is in logarithmic phase, trysinization is also collected in centrifuge tube, and the centrifugal 5min of 1000r/min, abandons supernatant.Then add lmL frozen storing liquid, dispel cell gently and make cell suspension, cell suspension is joined in the cryopreservation tube of 1.5mL.In the title of cryopreservation tube subscript clear-cells, shelf time, the name of preserver.First cryopreservation tube is put into 4 DEG C of refrigerators, place about 30min.Then be placed in-20 DEG C of refrigerators, place about 2h.Then be put in-80 DEG C of Ultralow Temperature Freezers to place and spend the night.Frozen cell was placed in the medium-term and long-term preservation of liquid nitrogen container in second day.
2, cell counting draws cell growth curve
(1) take the logarithm A375 melanoma cell in vegetative period, counting, adjustment cell density is 2 × 10
4/ mL.
(2) by cell suspension inoculation in 12 orifice plates, 1.5mL/ hole.
(3) after 12h, treat cell attachment, change 1mL plasma-free DMEM medium, and (compound (I) for DMSO dilution controls DMSO concentration below 1/1000 to use 5 μm of ol/L compounds (I) and DMSO respectively, guarantee cell harmless) process, often organize cell and establish 3 parallel holes, be placed in 5%C02,37 DEG C of constant incubators are cultivated.
(4) stop cultivating respectively at 0h, 24h, 48h, 72h, 96h, collect each group of cell, add 0.25% tryptic digestion, centrifugal, resuspended.
(5) cell counting: draw cell suspension 10 μ l, add isopyknic phenol blue area and divide viable cell, draw 10 μ L mixed solutions and add in cell counting count board groove gently, ensure tight and bubble.Under inverted microscope, the cell count in counting periphery four block plaid, substitutes into formula: archeocyte suspension concentration (cell count/mL)=(cell count/4 of 4 block plaid) × 10
4× extension rate.
(6) calculate number of viable cells, draw cell growth curve.
3, mtt assay detects cell proliferation
(1) the A375 cell of taking the logarithm vegetative period, digestion, centrifugal, resuspended, counting, in adjustment cell suspension, the density of cell is 2 × 10
4/ mL.
(2) by each group of cell suspension inoculation in 96 well culture plates, the 200 every holes of μ L, often organize cell 3 parallel holes, establish blank control wells (only adding substratum) simultaneously.
(3) 12h is after cell attachment, changes 200 μ L plasma-free DMEM medium, and uses 5 μm of ol/L compounds (I) and DMSO process respectively.
(4) stopped cultivating respectively at 0,1,2,3,4 day.
(5) add Methyl thiazoly tetrazolium assay (MTT) the 20 μ L that concentration is 5mg/mL to every hole and continue culturing cell 4h.
(6) inhale the substratum abandoned in each aperture, add 200 μ LDMSO, microoscillator is vibrated 10min, and crystallisate is fully dissolved.
(7) return to zero with blank control wells, automatic microplate reader is adopted to measure the absorption photometric value (OD value) at 570nm place, each hole, ability of cell proliferation is represented with the OD value of correspondence, the each group of mean value getting 3 parallel holes, take time as transverse axis, with each absorbance for the longitudinal axis draws cell growth curve.
4, BrdU Immunofluorescence test cell proliferation
(1) creep plate is prepared: creep plate is put into 75% alcohol and soak 24h and disinfect.
(2) aseptic creep plate is placed in 24 well culture plates, the A375 melanoma cell in vegetative period of taking the logarithm, digestion, centrifugal, make cell suspension, 2 × 10
4cell/ hole, is inoculated in and is placed with in the hole of creep plate.
(3) 12h is after cell attachment, changes 1mL plasma-free DMEM medium, and uses 5 μm of ol/L compounds (I) and DMSO process respectively.
(4) after 72h, add 10 μ lBrdU (1mg/mL) in the medium, cultivate 1h for 37 DEG C.
(5) take out 24 orifice plates, wash 5min × 2 time with cold PBS, 4% paraformaldehyde fixed cell, room temperature 30min.
(6) PBS washes 5min × 3 time.
(7) 2mol/LHCl process, after room temperature 10min, 37 DEG C of 20min.
(8) the penetrating process in cell 1%Triton ×-100, washes 5min × 3 time.
(9) 10% lowlenthal serums are closed, room temperature, lh.
(10) BrdU monoclonal antibody (1:300 dilution) is added, 37 DEG C of 1.5h.
(11) l × PBST shakes and washes 5min × 3 time.
(12) add BrdU bis-anti-(1:300), after room temperature lucifuge 1h, l × PBST washes 3 times.
(13) DAPI staining fluid transfect cell core 15min.
(14) PBS washes 5min × 3 time.
(15) 1 anti-fluorescent quenching mounting liquid is added, neutral gum mounting, fluorescence microscopy Microscopic observation, photograph.
5, statistical study
Application SPSS17.0 statistical analysis software, adopt two independent samples t test and one-way analysis of variance to carry out data analysis, data X ± S represents, P<0.05 is for there being statistical significance.
Three, result and conclusion
Cellular form is observed and counting display, and with DMSO control group ratio, after compound (I) process A375 cell 24h, 48h, 72h, viable count significantly reduces and reaches 44.5%.The results are shown in Table 1 (with control group ratio
*p<0.05,
*p<0.01, lower same).
MTT result shows, and compound (I) can significantly suppress A375 cell proliferation, and OD value is that the decline of concentration (5 μm of ol/L, 10 μm of ol/L, 20 μm of ol/L) dependency reaches 27.3%, and difference has statistical significance.The results are shown in Table 2.
Compound (I) energy check melanin tumor cell proliferation is confirmed further by BrdU immunofluorescence dyeing, 5 μm of ol/L compound (I) process A375 cells are after 3 days, compared with contrast (26.07 ± 2.59%), experimental group BrdU positive cell percentage (7.55 ± 2.76%) significantly reduces (P=0.000).The results are shown in Figure 3 (* * P<0.01).
This experiment have detected the impact that compound (I) is bred melanoma cell, cell counting and MTT analytical results all prove that compound (I) can remarkable check melanin tumor cell growth, and cell count and compound (I) concentration are that dependency declines.After BrdU immunofluorescence dyeing also shows compound (I) process, BrdU positive cell percentage is significantly lower than control group, illustrates that compound (I) can suppress A375 melanoma cell to be bred.Compound (I) may be melanomatous efficient targeting medicine.
Table 1 compound (I) dose-dependent inhibition A375 cell proliferation
(cell counting, unit: × l0
4/ ml) (
n=3)
| Group | 0h | 24h | 48h | 72h | 96h |
| DMSO | 2.87±0.23 | 4.72±0.30 | 7.36±0.13 | 7.56±0.39 | 8.04±0.27 |
| Compound (I)-5 μm of ol/L | 2.87±0.23 | 3.87±0.36* | 3.48±0.32** | 3.40±0.34** | 2.54±0.41** |
| Compound (I)-10 μm of ol/L | 2.87±0.23 | 2.36±0.28** | 2.95±0.37** | 1.56±0.36** | 0.89±0.26** |
Table 2 compound (I) dose-dependent inhibition A375 cell proliferation
(mtt assay, OD570) (
n=3)
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 1:9 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.
Claims (6)
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 dry leave of (a) sweet viburnum is pulverized, extract with 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), use 8% ethanol and 75% ethanol elution successively, collect 75% ethanol eluate, concentrating under reduced pressure obtains 75% ethanol elution thing medicinal extract; C in () step (b), 75% ethanol elution medicinal extract purification on normal-phase silica gel is separated, obtain 5 components successively with the methylene chloride-methanol gradient elution that volume ratio is 80:1,45:1,20:1,10:1 and 1:1; D in () step (c), component 3 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 25:1,20:1 and 15:1; 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 elutriant, elutriant concentrating under reduced pressure obtains pure compound (I).
3. preparation method according to claim 2, is characterized in that: described macroporous resin is AB-8 type macroporous adsorbent resin.
4. pharmaceutical composition, the compound according to claim 1 (I) wherein containing treatment significant quantity and pharmaceutically acceptable carrier.
5. the application of compound according to claim 1 (I) in the melanomatous medicine of preparation treatment.
6. the application of pharmaceutical composition according to claim 4 in the melanomatous medicine of preparation treatment.
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| CN109134184B (en) * | 2017-06-16 | 2021-03-19 | 中国科学院上海药物研究所 | Diterpenoid compounds, preparation method and application thereof |
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Application publication date: 20151118 |