CN105585609A - Aminophylline drug composition and medical application thereof - Google Patents
Aminophylline drug composition and medical application thereof Download PDFInfo
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07J—STEROIDS
- C07J73/00—Steroids in which the cyclopenta[a]hydrophenanthrene skeleton has been modified by substitution of one or two carbon atoms by hetero atoms
- C07J73/001—Steroids in which the cyclopenta[a]hydrophenanthrene skeleton has been modified by substitution of one or two carbon atoms by hetero atoms by one hetero atom
- C07J73/003—Steroids in which the cyclopenta[a]hydrophenanthrene skeleton has been modified by substitution of one or two carbon atoms by hetero atoms by one hetero atom by oxygen as hetero atom
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- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
- A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
- A61K31/519—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim ortho- or peri-condensed with heterocyclic rings
- A61K31/52—Purines, e.g. adenine
- A61K31/522—Purines, e.g. adenine having oxo groups directly attached to the heterocyclic ring, e.g. hypoxanthine, guanine, acyclovir
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- A61K31/56—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
- A61K31/58—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids containing heterocyclic rings, e.g. danazol, stanozolol, pancuronium or digitogenin
- A61K31/585—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids containing heterocyclic rings, e.g. danazol, stanozolol, pancuronium or digitogenin containing lactone rings, e.g. oxandrolone, bufalin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/73—Rosaceae (Rose family), e.g. strawberry, chokeberry, blackberry, pear or firethorn
- A61K36/734—Crataegus (hawthorn)
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Abstract
The invention discloses an aminophylline drug composition and medical application thereof. The provided aminophylline drug composition contains aminophylline and a natural product compound (I) which is separated from dry hawthorn fruit and is novel in structure, and the treatment effect on melanomata is poor when aminophylline and the natural product compound (I) act individually and is remarkably improved when aminophylline and the natural product compound (I) act in combination. The aminophylline drug composition can be developed into a drug for treating the melanomata, which has substantive features and remarkable progress in comparison with the prior art.
Description
Technical field
The invention belongs to biomedicine field, relate to the new purposes of aminophylline, be specifically related to a kind of aminophylline pharmaceutical composition andIt is to melanomatous therapeutic action.
Background technology
Aminophylline is theophylline and ethylenediamine double salt, and its pharmacological action is mainly from theophylline, and ethylenediamine makes its water-soluble enhancing. InsolubleIn methyl alcohol, ethanol, ether. This product has direct relexation to airway smooth muscle. Its mechanism of action more complicated, recognizes in the pastFor by suppressing phosphodiesterase, make due to the interior cAMP content raising of cell. Recently experiment thinks that the bronchiectasis of theophylline doesThat in addition, theophylline is purinoceptor retarding agent due to the result of endogenous adrenaline and norepinephrine release by part,Can resist the contraction to respiratory tract such as adenine. Theophylline can strengthen contraction of diaphragm power, especially effect in the time that contraction of diaphragm is unableMore remarkable, be therefore of value to and improve respiratory function. This product still has faint diastole coronary artery, and peripheral vascular and bile duct smooth muscle are doneWith. There are slight increase convergent force and slight diuresis.
Clinical practice mainly contains:
1. lax bronchial smooth muscle, the several kind of smooth muscle cells such as enteron aisle, biliary tract that also can relax, the hyperemia to tunica mucosa bronchiorum, oedemaAlso there is mitigation;
2. increase CO, expansion output and input renal arterioles, increase glomerular filtration rate(GFR and renal blood flow, suppresses far-endReabsorption sodium and chlorion;
3. increase the convergent force of Isolated Skeletal Muscular; In chronic obstructive pulmonary disease situation, improve myotility.
Melanoma, claims again malignant mela noma, is a class malignant tumour that derives from melanocyte, is common in skin, alsoSee the position such as mucous membrane, ocular choroid. Melanoma is the knurl kind that in skin neoplasin, grade malignancy is the highest, easily occurs turning at a distanceMove. Early diagnosis and therapy thereby seem particularly important.
Summary of the invention
The object of the present invention is to provide a kind of pharmaceutical composition of aminophylline, in this pharmaceutical composition, contain aminophylline and a kind of fromThe natural products that separates the novel structure obtaining in draft, aminophylline and this natural products can Synergistic treatment melanomas.
Above-mentioned purpose of the present invention is to be achieved by technical scheme below:
A kind of compound (I) with following structural formula,
A pharmaceutical composition for aminophylline, comprises aminophylline, compound as above (I) and acceptable pharmaceuticallyCarrier.
The preparation method of compound as above (I), comprises following operating procedure: (a) dry fruit of hawthorn pulverized,With 75～85% alcohol heat reflux extractions, merge extract, be concentrated into without alcohol taste, use successively benzinum, ethyl acetate and water saturationExtracting n-butyl alcohol, obtain respectively petroleum ether extract, acetic acid ethyl ester extract and n-butyl alcohol extract; (b) step (a)Middle n-butanol is got thing macroreticular resin removal of impurities, first uses 8 column volumes of 15% ethanol elution, then uses 12 posts of 70% ethanol elutionVolume, collects 70% eluent, and reduced pressure concentration obtains 70% ethanol elution concentrate; (c) 70% ethanol elution in step (b)Concentrate separates by purification on normal-phase silica gel, is the methylene chloride-methanol gradient elution of 70:1,35:1,15:1 and 5:1 successively by volume ratioObtain 4 components; (d) in step (c), component 4 use purification on normal-phase silica gel further separate, and are 8:1,5:1 successively by volume ratioObtain 3 components with the methylene chloride-methanol gradient elution of 2:1; (e) component 2 octadecylsilane key in step (d)The reverse phase silica gel closing separates, and the methanol aqueous solution isocratic elution that is 75% by concentration expressed in percentage by volume, collects 9～15 column volume wash-outsLiquid, eluent reduced pressure concentration obtains compound (I).
Further, in step (a), with 80% alcohol heat reflux extraction, merge extract.
Further, described macroreticular resin is D101 type macroporous absorbent resin.
The application of compound as above (I) in the melanomatous medicine of preparation treatment.
The application of the pharmaceutical composition of aminophylline as above in the melanomatous medicine of preparation treatment.
Advantage of the present invention:
In the pharmaceutical composition of aminophylline provided by the invention, contain aminophylline and a kind of novel structure of obtaining of separating from hawthornNatural products, when aminophylline and this natural products independent role, to melanomatous result for the treatment of a little less than; When the two synergy,Melanomatous result for the treatment of is significantly improved, can be developed to the melanomatous medicine for the treatment of. The present invention compared with prior artThere is outstanding substantive distinguishing features and significant progressive.
Detailed description of the invention
Further illustrate essentiality content of the present invention below in conjunction with embodiment, but do not limit protection domain of the present invention with this. To the greatest extentPipe is explained in detail the present invention with reference to preferred embodiment, and those of ordinary skill in the art should be appreciated that can be to the present inventionTechnical scheme modify or be equal to replacement, and do not depart from essence and the scope of technical solution of the present invention.
Embodiment 1: compound (I) separates preparation and structural identification
Reagent source: ethanol, benzinum, ethyl acetate, n-butanol, carrene are that analysis is pure, purchased from Shanghai Ling Feng chemistryReagent Co., Ltd, methyl alcohol, analyze pure, purchased from Jiangsu Han Bang chemical reagent Co., Ltd.
Separation method: (a) dry fruit of hawthorn (5kg) is pulverized, with 80% alcohol heat reflux extraction (20L × 3 time),Merge extract, be concentrated into without alcohol taste (4L), use successively benzinum (4L × 3 time), ethyl acetate (4L × 3 time) andWater saturated n-butanol (4L × 3 time) extraction, obtains respectively petroleum ether extract, acetic acid ethyl ester extract and extracting n-butyl alcoholThing; (b) D101 type macroreticular resin removal of impurities for acetic acid ethyl ester extract in step (a), first uses 8 posts of 15% ethanol elutionVolume, then use 12 column volumes of 70% ethanol elution, collect 70% eluent, reduced pressure concentration obtains 70% ethanol elution concentrate;(c) in step (b), 70% ethanol elution concentrate separates by purification on normal-phase silica gel, successively with volume ratio be 70:1 (8 column volumes),The methylene chloride-methanol gradient elution of 35:1 (8 column volumes), 15:1 (8 column volumes) and 5:1 (10 column volumes) obtainsTo 4 components; (d) in step (c), component 4 use purification on normal-phase silica gel further separate, and are 8:1 (8 posts successively by volume ratioVolume), the methylene chloride-methanol gradient elution of 5:1 (10 column volumes) and 2:1 (5 column volumes) obtains 3 components;(e) in step (d), the reverse phase silica gel of component 2 use octadecylsilane bondings separates, the first that is 75% by concentration expressed in percentage by volumeAlcohol solution isocratic elution, collects 9～15 column volume eluents, eluent reduced pressure concentration obtain compound (I) (295mg,HPLC normalization purity is greater than 98%).
Structural identification: yellow solid; HR-ESI-MS shows [M+H]+For m/z509.3259, can obtain molecule in conjunction with nuclear-magnetism featureFormula is C32H44O5, degree of unsaturation is 11. Proton nmr spectra data δH(ppm，DMSO-d6，500MHz)：H-1(6.06，d，J＝12.9Hz)，H-2(5.88，d，J＝12.9Hz)，H-4(4.44，m)，H-5(2.23，m)，H-6(0.78，m)，H-6(1.84，m)，H-7(1.14，m)，H-7(1.41，m)，H-8(1.78，dd，J＝10.5，6.7Hz)，H-11(1.50，m)，H-11(2.10，m)，H-12(1.63，m)，H-12(1.71，m)，H-15(1.28，m)，H-15(2.34，dd，J＝14.9，7.3Hz)，H-16(5.32，m)，H-16b(2.05，s)，H-17(2.51，m)，H-18(1.08，s)，H-19(0.89，d，J＝4.3Hz)，H-19(0.98，d，J＝4.3Hz)，H-21(1.81，s)，H-22(6.61，s)，H-24aa(6.04，s)，H-24ab(5.85，s)，H-25(2.88，m)，H-26(1.03，d，J＝6.6Hz)，H-27(1.11，d，J＝6.6Hz)，H-28(1.27，d，J＝6.7Hz)，H-30(0.99，S); Carbon-13 nmr spectra data δC(ppm，DMSO-d6，125MHz)：150.2(CH，1-C)，119.6(CH，2-C)，168.5(C，3-C)，79.3(CH，4-C)，44.8(CH，5-C)，23.4(CH2，6-C)，24.8(CH2，7-C)，46.2(CH，8-C)，30.2(C，9-C)，37.0(C，10-C)，28.5(CH2，11-C)，32.6(CH2，12-C)，45.2(C，13-C)，47.3(C，14-C)，46.9(CH2，15-C)，77.1(CH，16-C)，170.2(C，16a-C)，22.3(CH3，16b-C)，59.5(CH，17-C)，18.5(CH3，18-C)， 32.5(CH2，19-C)，162.4(C，20-C)，16.8(CH3，21-C)，126.6(CH，22-C)，194.8(C，23-C)，154.3(C，24-C)，119.8(CH2，24a-C)，29.5(CH，25-C)，22.1(CH3，26-C)，21.8(CH3，27-C)，18.8(CH3，28-C)，20.1(CH3, 30-C). 1715cm-in IR spectrogram1With 1694cm-1Absorption band shows to exist in this compound structure carbonyl and conjugation carbonyl; Absorption maximum 207nm in UV spectrogramWith in 245nm description architecture, there is conjugated double bond. Hydrogen spectrum nuclear magnetic data shows: in this structure, degree of unsaturation is two by 3 C-CBond structure, 3 carbonyl structures and 5 cyclic skeleton compositions. δH0.89 and δH0.98 coupling constant is 4.3Hz, shows thisThere is cyclopropane structure in compound. Carbon spectrum nuclear magnetic data and DEPT spectrum data show: in this compound structure, contain 32 carbon,Wherein there are seven methyl (δC22.3,18.5,16.8,22.1,21.8,18.8 and 20.1), seven methylene (δC23.4、24.8、28.5,32.6,46.9,32.5 and 119.8), nine methine (δC150.2、119.6、79.3、44.8、46.2、77.1、59.5,126.6 and 29.5), nine tertiary carbon (δC168.5、30.2、37.0、45.2、47.3、170.2、162.4、194.8With 154.3), a ketone group (δC194.8), two ester carbonyl group (δC168.5 and 170.2), two oxygen carbon (δ of companyC79.3With 77.1), six olefinic carbon (δC150.2 and 119.6,162.4 and 126.6; 154.3 with 119.8). Resolve by HMBCKnown δC170.2 carbonyl carbon and δH2.05 H-16b methyl composition acetyl group, δ simultaneouslyH5.32 H-16 and δC170.2 carbonylAlso there is correlation in carbon, shows that C-16 position is connected with an acetoxyl group. Compose Data Analysis δ by NOESYH4.44 H-4,δH1.27 H-28 and δH2.23 H-5 and δH1.84 H-6 relation can illustrate 28-methyl Relative configuration. Comprehensive hydrogenSpectrum, carbon spectrum, HMBC spectrum and NOESY spectrum, and document is about correlation type nuclear magnetic data, can substantially determine this compoundAs follows, spatial configuration is further tested and is determined by ECD, and theoretical value and experiment value are basically identical.
Embodiment 2: to melanomatous therapeutic action
One, material and instrument
People A375 melanoma cell strain is given by Third Military Medical University. Compound (I) self-control, preparation method is shown in embodiment 1,HPLC normalization purity is greater than 98%. DMEM culture medium, 0.25% pancreatin are purchased from Gibco company of the U.S.. MTT (3-(4,5-Dimethylthiazole-2)-2,5-diphenyl tetrazole bromine salt; Tetramethyl azo azoles indigo plant), DMSO (dimethyl sulfoxide (DMSO)), BrdU purchased fromSigma company of the U.S.. Mouse-anti BrdU polyclonal antibody is purchased from American AB cam company. Mountain sheep anti mouse two is anti-purchased from U.S. CellSignaling company. AnnexinV-FITC/PI cell apoptosis detection kit is purchased from Promega company of the U.S..
Constant temperature CO2Incubator, general refrigerator ,-80 degree refrigerators are Forma company of U.S. product. Superclean bench (ChinaSuzhou cleaning project company). Inverted microscope, inverted fluorescence microscope (olympus company). Electro-heating standing-temperature cultivator (inShanghai leap medical apparatus and instruments factory of state). Automatically ELIASA (Japanese Wako company). Uv-spectrophotometric instrument (U.S. BeckmanCompany). J6-HC supercentrifuge (Beckman company of the U.S.). Low-temperature trace centrifuge (German Eppendorf company).Vibration shaking table (Forma company of the U.S.).
Two, test method
1, cell is cultivated
1.1 cell recovery
Frozen A375 MC in liquid nitrogen container is taken out, put into rapidly the warm water of 37 DEG C, shake makes it gentlyMelt as early as possible (approximately lmin). Then sucking-off cell suspension, joins in the centrifuge tube that is added with 2mL culture medium, gentlyPiping and druming mixes, 800r/min, and centrifugal 5min, discards upper strata culture medium. With the mould G/ streptomysin that contains 10%FBS and 1%DMEM culture medium 8mL make after cell suspension, be seeded in 10cm culture plate. Then be placed in 5%CO2, 37 DEG C of perseverancesIn temperature incubator, cultivate.
1.2 passage
When under microscope, observation of cell degrees of fusion reaches 80%-90%, can go down to posterity. First use 2mLPBS washed cell 2 times, soAfter add 0.25% pancreatin lmL. Level shake culture plate, makes digestive juice can cover the surface of cell gently, is then placed in 37 DEG CIn incubator, digest about lmin. Be placed under microscope and see that cell rounding bounces back, the culture medium that then adds rapidly 1mL to contain FBSStop digestion. Blow and beat gently to cell and be uniformly dispersed, then go down to posterity according to 1:2 or 1:3 according to cell density, be re-seeded intoIn new culture plate, be placed in 5%CO2, cultivate in 37 DEG C of constant incubators.
1.3 cell cryopreservation
Get the cell of 1 dish in exponential phase, trypsinization is also collected in centrifuge tube, and the centrifugal 5min of 1000r/min, abandonsSupernatant. Then add lmL cryopreserving liquid, dispel gently cell and make cell suspension, cell suspension is joined to freezing of 1.5mLDeposit in pipe. In the title of cryopreservation tube subscript clear-cells, holding time, preserver's name. 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 ultra low temperature freezers and place and spend the night. TheFrozen cell was placed in to the medium-term and long-term preservation of liquid nitrogen container in two days.
2, cell counting is drawn cell growth curve
(1) the A375 MC in growth period of taking the logarithm, counting, adjusting cell density is 2 × 104/mL。
(2) by cell suspension inoculation in 12 orifice plates, 1.5mL/ hole.
(3) after 12h, treat cell attachment, change 1mL serum-free DMEM culture medium, and use respectively 5 μ mol/L aminophyllines,5 μ mol/L compounds (I), 2.5 μ mol/L aminophyllines and 2.5 μ mol/L compound (I) compositions, DMSO[forThe medicine of DMSO dilution, controls DMSO concentration below 1/1000, guarantees cell harmless] to process, every group of cell establishes 3Individual parallel hole, is placed in 5%CO2, cultivate in 37 DEG C of constant incubators.
(4) stop cultivating respectively at 0h, 24h, 48h, 72h, collect each group of cell, add 0.25% Trypsin Induced, fromThe heart, resuspended.
(5) cell count: draw cell suspension 10 μ L, add isopyknic phenol blue area to divide living cells, draw 10 μ L mixedClose liquid and add gently in cell counting count board groove, ensure tight and bubble. Under inverted microscope, four large grids of counting peripheryInterior cell number, substitution formula: archaeocyte suspension concentration (cell number/mL)=(cell number/4 of 4 large grids) × 104×Extension rate.
(6) calculate living cells number, draw cell growth curve.
3, mtt assay detects cell proliferation
(1) the A375 cell of taking the logarithm growth period, digestion, centrifugal, resuspended, counting, adjusts cell in cell suspensionDensity is 2 × 104/mL。
(2) by each group of cell suspension inoculation in 96 well culture plates, the 200 every holes of μ L, 3 parallel holes of every group of cell, simultaneouslyIf blank hole (only adding culture medium).
(3) 12h, after cell attachment, changes 200 μ L serum-free DMEM culture mediums, and use respectively 5 μ mol/L aminophyllines,5 μ mol/L compounds (I), 2.5 μ mol/L aminophyllines and 2.5 μ mol/L compound (I) compositions, DMSO process.
(4) stopped cultivating respectively at 0,1,2,3,4 day.
(5) adding concentration to every hole is tetramethyl azo azoles indigo plant (MTT) the 20 μ L continuation cultured cell 4h of 5mg/mL.
(6) inhale the culture medium of abandoning in each aperture, add 200 μ LDMSO, the 10min that vibrates in microoscillator, makes crystallizationThing fully dissolves.
(7) with the zeroing of blank hole, adopt automatic ELIASA to measure the absorption photometric value at 570nm place, each hole (OD value),With corresponding OD value representation ability of cell proliferation, get the mean value of 3 parallel holes for each group, taking the time as transverse axis, with each extinctionDegree value is that the longitudinal axis is drawn cell growth curve.
4, statistical analysis
Application SPSS18.0 statistical analysis software, adopts two independent sample t inspections and one-way analysis of variance to carry out data and dividesAnalyse, data represent with X ± S, and P < 0.05 is for there being statistical significance.
Three, result and conclusion
1, cellular morphology is observed and count results
Cellular morphology is observed and counting shows, after aminophylline and compound (I) compositions-treated A375 cell 48h, 72h,Viable count significantly reduces, and inhibition is significantly better than aminophylline or compound (I) independent role 48h or 72h, difference toolThere is statistical significance (P < 0.05).
The results are shown in Table 1.
Impact (cell counting, the unit: × l0 of table 1 on A375 cell proliferation4/mL)(n＝3)
| Group | 0h | 24h | 48h | 72h |
| DMSO group | 2.89±0.32 | 4.82±0.30 | 7.46±0.13 | 7.66±0.39 |
| Aminophylline group | 2.89±0.32 | 3.97±0.36 | 3.58±0.32 | 3.12±0.34 |
| Compound (I) group | 2.89±0.32 | 3.92±0.39 | 3.53±0.35 | 3.11±0.29 |
| Aminophylline and compound (I) composition group | 2.89±0.32 | 2.46±0.28 | 2.15±0.37 | 1.66±0.36 |
2, mtt assay experimental result
MTT result shows, aminophylline or compound (I) independent role, aminophylline and compound (I) combination of compositions workCan suppress A375 cell proliferation with equal, and inhibition when aminophylline and compound (I) compound action is significantly better than ammonia teaThe inhibition (P < 0.05) of alkali or compound (I) independent role.
The results are shown in Table 2.
The inhibition (mtt assay, OD570) of table 2 to A375 cell proliferation (n＝3)
| Group | 0h | 24h | 48h | 72h |
| DMSO group | 0.183±0.011 | 0.286±0.033 | 0.498±0.012 | 0.498±0.017 |
| Aminophylline group | 0.183±0.011 | 0.198±0.019 | 0.193±0.014 | 0.175±0.023 |
| Compound (I) group | 0.183±0.011 | 0.199±0.037 | 0.195±0.024 | 0.172±0.029 |
| Aminophylline and compound (I) composition group | 0.183±0.011 | 0.098±0.005 | 0.064±0.002 | 0.035±0.004 |
Result of the test shows, the composition of aminophylline and compound (I) can significantly suppress the propagation of A375 cell, and it presses downMake of being significantly higher than the inhibition to A375 cell separately of aminophylline or compound (I). Aminophylline and compound (I) itBetween exist synergy.
Those of ordinary skill in the art should be appreciated that and can modify or be equal to replacement technical scheme of the present invention, andDo not depart from essence and the protection domain of technical solution of the present invention.
Claims (7)
1. one kind has the compound (I) of following structural formula,
2. a pharmaceutical composition for aminophylline, is characterized in that: comprise aminophylline, compound as claimed in claim 1 (I)Pharmaceutically acceptable carrier.
3. the preparation method of compound claimed in claim 1 (I), is characterized in that, comprises following operating procedure: (a)The dry fruit of hawthorn is pulverized, with 75～85% alcohol heat reflux extractions, merged extract, be concentrated into without alcohol taste, use successively stoneOil ether, ethyl acetate and water saturated extracting n-butyl alcohol, obtain respectively petroleum ether extract, acetic acid ethyl ester extract and n-butanolExtract; (b) in step (a), n-butanol is got thing macroreticular resin removal of impurities, first uses 8 column volumes of 15% ethanol elution, thenWith 12 column volumes of 70% ethanol elution, collect 70% eluent, reduced pressure concentration obtains 70% ethanol elution concentrate; (c) stepSuddenly in (b), 70% ethanol elution concentrate separates by purification on normal-phase silica gel, is 70:1,35:1,15:1 and 5:1 successively by volume ratioMethylene chloride-methanol gradient elution obtains 4 components; (d) in step (c), component 4 use purification on normal-phase silica gel further separate, and comply withInferiorly obtain 3 components with the methylene chloride-methanol gradient elution that volume ratio is 8:1,5:1 and 2:1; (e) group in step (d)Divide the reverse phase silica gel of 2 use octadecylsilane bondings to separate, the methanol aqueous solution isocratic elution that is 75% by concentration expressed in percentage by volume,Collect 9～15 column volume eluents, eluent reduced pressure concentration obtains compound (I).
4. the preparation method of compound according to claim 3 (I), is characterized in that: in step (a), with 80%Alcohol heat reflux extracts, and merges extract.
5. the preparation method of compound according to claim 3 (I), is characterized in that: described macroreticular resin is D101Type macroporous absorbent resin.
6. the application of compound claimed in claim 1 (I) in the melanomatous medicine of preparation treatment.
7. the application of the pharmaceutical composition of aminophylline claimed in claim 2 in the melanomatous medicine of preparation treatment.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106496205A (en) * | 2016-09-12 | 2017-03-15 | 南通市科通科技信息咨询有限公司 | A kind of pharmaceutical composition of lavo-ofloxacin and its medical usage |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104961791A (en) * | 2015-06-05 | 2015-10-07 | 中国科学院西北高原生物研究所 | Novel triterpene compound in potentilla anserina, and preparation method and application thereof |
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| CN104961791A (en) * | 2015-06-05 | 2015-10-07 | 中国科学院西北高原生物研究所 | Novel triterpene compound in potentilla anserina, and preparation method and application thereof |
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| CN106496205A (en) * | 2016-09-12 | 2017-03-15 | 南通市科通科技信息咨询有限公司 | A kind of pharmaceutical composition of lavo-ofloxacin and its medical usage |
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