CN107540687A - A kind of new phloroglucinol derivatives compound and preparation method thereof and medical usage - Google Patents

Abstract

The invention discloses a kind of new phloroglucinol derivatives compound and preparation method thereof and medical usage.The compound is a kind of novel phloroglucinol derivatives compound of structure, extracting and developing can purify to obtain from the dry mature kernal of hickory nut to report first.In vitro test proves that the compound is inhibited to human brain malignant glioblastoma U251 cells, and inhibitory action in time, dose dependent, i.e. the compound concentration is higher, and action time is longer, its inhibitory action is stronger, can be used for developing into the medicine for the treatment of glioma.

Description

A kind of new phloroglucinol derivatives compound and preparation method thereof and medical usage

Technical field

The invention belongs to technical field of pharmaceuticals, and in particular to isolated one kind from the dry mature kernal of hickory nut Phloroglucinol derivatives compound with treatment glioma effect and preparation method thereof.

Background technology

Hickory nut (Carya cathayensis) is that Juglandaceae (Juglandaceae) hickory (Carya) fallen leaves are tall Wood, alias Chinese walnut, numb walnut, the plant whole world of hickory share 18 kinds, 2 subspecies, China be its original producton location it One.Hickory nut is distributed in China north and south, Yunnan, Hunan, Guizhou, the main producing region that zhejiang and other places area is China hickory nut.Mountain Walnut is sweet, mild-natured, first recorded in 3rd century of Christian era Shanxi for written by Zhang Hua《Natural science will》, rear Tao Hongjing《Mingyi Bielu》During with Lee Precious《Compendium of Materia Medica》Also it is on the books.It has invigorating qi and benefiting blood, adjusts dry resolving sputum, temperature compensation kidney lung, removing toxicity for detumescence and other effects.Hickory nut Whole body is all precious, and walnut kernel can nourish blood vessels, improve a poor appetite, pitch-black beard and hair, regulation neurasthenia, amnesia；Fresh quench skin is decocted Soup embathes, and controls pin hemorrhoid；Fresh exocarp, which is smash, takes juice, and diseases such as skin tinea etc. are controlled in wiping.

Hickory nut contains the chemical compositions such as flavones, quinones, phenolic acid class.Modern study shows that hickory nut has antitumor, suppression Bacterium, anti-oxidant etc. act on.

The content of the invention

It is an object of the invention to provide one kind isolated in a kind of dry mature kernal from hickory nut to have treatment Phloroglucinol derivatives compound of glioma effect and preparation method thereof.

The above-mentioned purpose of the present invention is achieved by following technical scheme：

Compound (I) with following structural formula,

The preparation method of described compound (I), includes following operating procedure：(a) by the dry mature kernal of hickory nut Crush, with 75~85% alcohol heat reflux extract, merge extract solution, be concentrated into no alcohol taste, successively with petroleum ether, ethyl acetate and Water saturated extracting n-butyl alcohol, respectively obtain petroleum ether extract, acetic acid ethyl ester extract and n-butyl alcohol extract；(b) step (a) acetic acid ethyl ester extract is cleaned with macroreticular resin in, first with 15% ethanol elution, 8 column volumes, then with 75% ethanol elution 10 column volumes, 75% ethanol eluate is collected, be concentrated under reduced pressure to obtain 75% ethanol elution thing medicinal extract；(c) 75% second in step (b) Alcohol elution medicinal extract is separated with purification on normal-phase silica gel, is successively 85 with volume ratio:1、45:1、20:1、10:1 and 1:1 dichloromethane-first Alcohol gradient elution obtains 5 components；(d) component 3 is further separated with purification on normal-phase silica gel in step (c), is successively 25 with volume ratio: 1、20:1 and 10:1 methylene chloride-methanol gradient elution obtains 3 components；(e) the octadecyl silicon of component 2 in step (d) The reverse phase silica gel separation of alkane bonding, with the methanol aqueous solution isocratic elution that concentration expressed in percentage by volume is 75%, collects 8~10 cylinders Product eluent, eluent are concentrated under reduced pressure to give pure compound (I).

Further, in step (a), extracted with 80% alcohol heat reflux, merge extract solution.

Further, the macroreticular resin is AB-8 type macroporous absorbent resins.

A kind of pharmaceutical composition, wherein described compound (I) and pharmaceutically acceptable load containing therapeutically effective amount Body.

Application of the described compound (I) in the medicine for preparing treatment glioma.

Application of the described pharmaceutical composition in the medicine for preparing treatment glioma.

It when the compounds of this invention is used as medicine, can directly use, or be used in the form of pharmaceutical composition.

The pharmaceutical composition contains the compounds of this invention (I) of therapeutically effective amount, and remaining is pharmaceutically acceptable, right The nontoxic and inert pharmaceutical acceptable carrier of humans and animals and/or excipient.

Described pharmaceutical acceptable carrier or excipient is one or more selected from solid, semisolid and liquid diluent, filler And pharmaceutical preparation assistant agent.The pharmaceutical composition of the present invention is used in the form of per weight dose.Medicine of the present invention can The patient for needing to treat is applied to by oral or injection form.For it is oral when, tablet, sustained release tablets, control can be made into Release piece, capsule, dripping pill, micropill, supensoid agent, emulsion, powder or granule, oral liquid etc.；During for injecting, sterilizing can be made into Water-based or oily solution, aseptic powder injection, liposome or emulsion etc..

Brief description of the drawings

Fig. 1 is compound (I) structural formula；

Fig. 2 is the theoretical ECD values of compound (I) compared with testing ECD values；

Fig. 3 is the change of U251 cells hole apoptosis rate after compound (I) processing.

Embodiment

The essentiality content of the present invention is further illustrated with reference to embodiment, but present invention protection model is not limited with this Enclose.Although being explained in detail with reference to preferred embodiment to the present invention, it will be understood by those within the art that, can be right Technical scheme is modified or equivalent substitution, without departing from the spirit and scope of technical solution of the present invention.

Embodiment 1：Compound (I) separation prepares and structural identification

Reagent source：Ethanol, petroleum ether, ethyl acetate, n-butanol, dichloromethane are pure to analyze, and peaking is insulted purchased from Shanghai Reagent Co., Ltd is learned, methanol, analysis is pure, purchased from Jiangsu Han Bang chemical reagent Co., Ltd.

Preparation method：(a) hickory nut dry mature kernal (10kg) crush, with 80% alcohol heat reflux extract (25L × 3 times), merge extract solution, no alcohol taste (3L) is concentrated into, successively with petroleum ether (3L × 3 time), ethyl acetate (3L × 3 time) and water N-butanol (3L × 3 time) extraction of saturation, respectively obtain petroleum ether extract, acetic acid ethyl ester extract (398g) and n-butanol extraction Take thing；(b) acetic acid ethyl ester extract is cleaned with AB-8 types macroreticular resin in step (a), first with 15% ethanol elution, 8 cylinders Product, then with 75% ethanol elution, 10 column volumes, 75% ethanol eluate is collected, be concentrated under reduced pressure to obtain 75% ethanol elution thing medicinal extract (163g)；(c) 75% ethanol elution medicinal extract is separated with 200-300 mesh purification on normal-phase silica gel in step (b), is successively 85 with volume ratio:1 (10 column volumes), 45:1 (9 column volumes), 20:1 (8 column volumes), 10:1 (8 column volumes) and 1:1 (5 column volumes) Methylene chloride-methanol gradient elution obtain 5 components；(d) component 3 (49g) 200-300 mesh purification on normal-phase silica gel in step (c) Further separation, it is successively 25 with volume ratio:1 (8 column volumes), 20:1 (10 column volumes) and 10:1 (5 column volumes) Methylene chloride-methanol gradient elution obtains 3 components；(e) in step (d) component 2 (31g) be bonded with octadecylsilane it is anti- Phase silica gel ODS-C18 is separated, and with the methanol aqueous solution isocratic elution that concentration expressed in percentage by volume is 75%, collects 8~10 column volumes Eluent, eluent are concentrated under reduced pressure to give pure compound (I) (244mg).

Structural identification：Yellow jelly；HR-ESIMS is shown [M+Na]⁺, can with reference to nuclear-magnetism feature for m/z 539.2822 It is C to obtain molecular formula₃₃H₄₀O₅, degree of unsaturation 14.Hydrogen nuclear magnetic resonance modal data δ_H(ppm, DMSO-d₆, 400MHz)：H-7 (2.53, t, J=13.0), H-7 (2.45, m), H-8 (2.31, m), H-10 (1.78, dd, J=12.1,7.2), H-10 (1.32, M), H-11 (2.04, m), H-11 (1.56, m), H-12 (1.89, m), H-14 (2.40, d, J=14.2), H-14 (2.22, d, J =14.2), H-15 (1.28, s), H-16 (1.37, s), H-17 (2.99, dd, J=13.2,9.9), H-17 (2.93, dd, J= 13.2,7.7), H-18 (5.38, t, J=7.3), H-20 (1.67, s), H-21 (1.59, s), H-22 (2.81, dd, J=12.7, 8.8), H-22 (2.69, dd, J=12.7,6.8), H-23 (5.09, t, J=7.3), H-25 (1.46, s), H-26 (1.48, s), H-29 (7.87, br, d, J=5.2), H-30 (7.36, t, J=7.2), H-31 (7.39, t, J=6.9), H-32 (7.36, t, J =7.2), H-33 (7.87, br, d, J=5.2)；Carbon-13 nmr spectra data δ_C(ppm, DMSO-d₆, 100MHz)：192.1 (C, 1-C), 112.7 (C, 2-C), 194.2 (C, 3-C), 60.8 (C, 4-C), 215.1 (C, 5-C), 60.3 (C, 6-C), 29.4 (CH₂, 7-C), 43.1 (CH, 8-C), 78.2 (C, 9-C), 40.5 (CH₂, 10-C), 22.7 (CH₂, 11-C), 44.2 (CH, 12-C), 70.3 (C, 13-C), 38.7 (CH₂, 14-C), 27.9 (CH₃, 15-C), 23.6 (CH₃, 16-C), 35.0 (CH₂, 17-C), 117.9 (CH, 18-C), 136.4 (C, 19-C), 25.6 (CH₃, 20-C), 17.5 (CH₃, 21-C), 39.7 (CH₂, 22-C), 116.2 (CH, 23- C), 137.6 (C, 24-C), 25.4 (CH₃, 25-C), 17.3 (CH₃, 26-C), 198.7 (C, 27-C), 137.8 (C, 28-C), 127.5 (CH, 29-C), 126.8 (CH, 30-C), 131.7 (CH, 31-C), 126.9 (CH, 32-C), 127.6 (CH, 33-C)；Carbon Atomic tag is referring to Fig. 1.¹H H NMR spectroscopies show six methyl signals (δ H1.28,1.37,1.46,1.48,1.59,1.67), two Individual olefinic protons (δ H5.09 and 5.38), five aromatic protons are by (δ H7.36,7.36,7.39,7.87,7.87).¹³C H NMR spectroscopies 33 carbon signals of display, including six methyl, six methylene, nine methines (seven olefinic methines), and 12 Quaternary carbon (three ketone carbonyls, four olefinic quaternary carbons, two oxygen-containing quaternary carbons, and an oxygen-containing quaternary carbon of olefinic).H in HMBC spectrums₂-7 And H₂- 14 and C-3, C-4 and C-5, Me-15 and C-12, C-13 and C-14, Me-16 and C-8, C-9 and C-10, and H₂- 17 and H₂- 22 and C-1, C-5 and C-6 correlation show that the compound is phloroglucinol derivatives compound.In HMBC spectrums, H-17 and H-22 With C-1, C-5 and C-6, Me-20 and Me-21 and C-19 and C-18, and Me-25 and Me-26 and C-24 and C-23 correlation Show that C-6 positions are connected with two 2- methyl-2-butene groups.According to two oxygen-containing carbon signal [δ C70.3 (C-13) and 78.2 (C- 9)], and nuclear magnetic data and degree of unsaturation, it is known that an oxygen bridge between C-9 and C-13 be present.According to nuclear magnetic data, and H-29 (δ H7.87, br, d, J=5.2Hz) and carbonyl carbon signals C-27 (δ C198.7) correlation shows monosubstituted in HMBC spectrums Phenyl ring is connected with C-27.Me-15 and H-8 correlation shows that Me-15 and H-8 is beta comfiguration in NOESY spectrums.In addition H-12 and H-8 And Me-16, Me-15 and H-14 β, H-8 and H-7 β correlation show that ring B and C are cis fusion, and H-12 and Me-16 is β structures Type.H-18 and H-7 β, H-23 and H-14 β, and H-29 and H-14 α correlation show the sequence of C-5 and C-6 in A rings in spiral shell The upside of carbon (C-4).Comprehensive hydrogen spectrum, carbon spectrum, HMBC spectrums and NOESY spectrums, and document is on correlation type nuclear magnetic data, can base For this determination compound as shown in figure 1, spatial configuration further tests determination by ECD, theoretical value is basically identical with experiment value (Fig. 2).

Embodiment 2：Compound (I) pharmacological action is tested

First, material and instrument

This experiment uses human brain malignant glioblastoma U251 cell lines, purchased from American Type Culture Collection center. DMEM high glucose mediums are purchased from Hyclone companies of the U.S.；Sodium chloride, sodium hydroxide, potassium chloride, chlorination oxygen, sodium hydroxide, phosphoric acid Disodium hydrogen, sodium dihydrogen phosphate, potassium dihydrogen phosphate, methanol are purchased from Nanjing chemical plant.Compound (I) is made by oneself, HPLC normalization purity More than 98%.Glycine, Tris, Tween20, dimethyl sulfoxide (DMSO) (DMEM) are purchased from Sigma Co., USA；Aimexin-V/PI is thin Bao Coverlet die kit and are purchased from Invitrogen companies of the U.S.；Cell pyrolysis liquid, 0.25% trypsase, Cell Counting Kit-8 kits, penicillin, streptomysin, Hoechst33258, nucleus dyestuff DAPI, PMSF, Western blot gel are matched somebody with somebody Kit processed, Nuclear extract extracts kit, coomassie brilliant blue staining liquid, standard protein, 5X protein denaturation buffer solutions box, The super quick luminescent solutions of ECL, developing fixing kit, X-ray film, nitrocellulose filter, anti-fluorescent quenching mounting liquid are green purchased from Jiangsu Skies biology grinds the institute that makes internal disorder or usurp.Hyclone is purchased from Hangzhou Chinese holly biotechnology company；Rabbit-anti people P65 antibody is purchased from U.S. NOVUS Company；Rabbit-anti people HistonH3 antibody is purchased from CST companies of the U.S.；Goat antirabbit HRP secondary antibodies are purchased from SantaCruz companies of the U.S.； Goat antirabbit fluorescence secondary antibody is purchased from EarthOx companies of the U.S..

- 20 DEG C of low temperature refrigerators (Haier Qingdao), -80 DEG C of low temperature refrigerators (Haier Qingdao), low temperature refrigerator (Haier Qingdao) are high Fast centrifuge (flying crane Shanghai), culture dish/plate (the Coming U.S.), centrifuge tube (the Coming U.S.), cell culture incubator (Thermo The U.S.), inverted microscope (Olympus Japan), fluorescence inverted phase contrast microscope (ZEISS Germany), multipurpose thermostatic water bath (leap Shanghai), electronic balance instrument (Shanghai secret scientific instrument), superclean bench (ARITECH Japan), micropipettor (Dragon Finland), ELIASA (Bio-RAD Germany), flow cytometer (BDAccuriC6

The U.S.).

2nd, test method

1st, cell culture and passage

People's glioblastoma cell line U251 culture is containing 10% hyclone, 1% penicillin, 1% streptomysin In DMEM high glucose mediums, in 37 DEG C, 5%CO₂Cellar culture in incubator, replacing fresh culture was given every 2 days.Treat cell After being fused to 80% or so, culture medium is absorbed, and is washed 3 times with the Sterile phosphate phthalate buffer of preheating, then into culture dish Add the cell dissociation buffer 1mL containing 0.25% race's protease and 0.02%EDTA to digest about 2 minutes, light Microscopic observation cell can See that gradually retraction is rounded cell, intercellular spaces broaden, and now remove cell dissociation buffer, add complete medium 4mL and terminate digestion, Softly cell suspension is made in piping and druming to 1mL pipette tips, and low speed centrifuge 1000rpm is centrifuged 5 minutes, absorbs supernatant, adds training completely Support base to be resuspended, after soft piping and druming uniformly, rule of thumb by 1:3~1:5 passages, the cell after passage, which is placed in incubator, to be continued Training, is about passaged twice a week, to keep cell to be in exponential phase.Method for cell count：Cell counting count board wiped clean, and Cover glass is covered on tally；Cell suspension is slowly dropped into tally along cover glass edge, light Microscopic observation, cell is saturating Bright, refractivity is living cells well, counts the viable count in 4 block plaids.Viable count is put down in every cell number=each grid Average × extension rate × 10⁴。

2nd, CCK-8 detects cells survival rate

Take the logarithm the U251 cells in growth period, single cell suspension is made with the 0.25% pancreatin digestion piping and druming cell containing EDTA And count, adjust cell concentration about 2 × l0 with complete medium⁴Individual/mL, cell is inoculated in 96 well culture plates, per hole 100 μ L, it is placed in 37 DEG C, 5%CO₂Cultivated 12 hours in incubator, absorb former culture medium, different disposal is given by packet：Blank control Group (equivalent complete medium), compound (I) group sets 1,2,4mmol/L, every group sets three repeating holes, sterile PBS closed perimeters Each hole, it is again placed in 37 DEG C, 5%CO₂In incubator, 96 orifice plates are taken out after cultivating 24,48,72 hours respectively, it is former to absorb each group There is culture medium, be replaced by 100 μ L DMEM nutrient solutions, background group is acellular to only have DMEM nutrient solutions.10 μ L CCK-8 are added per hole Solution, it is placed in incubator and continues culture 2 hours.Determined with full-automatic ELIASA per hole absorbance (OD values), wavelength setting At 450mn, the average of every group of multiple holes is taken.Above-mentioned experimental procedure is repeated 3 times, and calculates average value.Cells survival rate calculation formula It is as follows:Cells survival rate=(experimental group OD values-background group OD values)/(control group OD values-background group OD values) × 100%, draw Growth inhibition curve map, and the credit analysis that takes statistics.

3rd, Hoechst 33258 is dyed

Experiment uses green skies company Hoechst33258 dyeing liquors, during apoptosis, can be appreciated that apoptosis Nucleus is in fine and close dense dye, or in the fine and close dense dye of chunky shape.According to cells survival rate result, choose 2mmol/L compounds (I) and make The condition dyed by the use of 48 hours as Hoechst 33258.By cell dissociation, centrifugation, counting, planted after adjusting cell concentration into 24 In orifice plate, 37 DEG C, 5%CO are placed in₂Cultivated 12 hours in incubator, absorb original culture medium, different disposal is given by packet:It is empty White control group (equivalent complete medium), compound (I) group sets 1,2,4mmol/L, sterile each hole of PBS closed perimeters, put again In 37 DEG C, 5%CO₂In incubator, 24 orifice plates are taken out after continuing culture 48 hours, absorb the original culture medium of each group, sterile PBS is washed 3 times, 0.5mL polies formic acid is added per hole and fixes cell 15 minutes, PBS is washed 3 times, and each hole adds 100 μ L Hoechst 33258 Dyeing liquor, 10min being dyed under room temperature dark surrounds, removing dyeing liquor, PBS shaking tables rock flushing 3 times, 5 minutes every time, wash most liquid Body, the anti-fluorescent quenching liquid of drop is added dropwise per hole, being taken pictures under the fluorescence microscope of 340nm wavelength and observing Cell Image Analyzer becomes Change.

3rd, Flow cytometry cell, which follows, dies

According to cells survival rate result, 48 hours particular point in times died as detection cell pillbox are chosen.Take and be in logarithm Growth period and U251 cells in good condition, routinely had digestive transfer culture, is inoculated in 37 DEG C, 5%CO in 6 orifice plates₂Incubated in incubator 12h is educated, different disposal is given by packet：Blank control group (equivalent complete medium), compound (I) group sets 1,2,4mmol/L, It is placed in 37 DEG C, 5%CO₂After continuing culture in incubator 48 hours, original nutrient solution is washed till in 5mL centrifuge tubes, PBS washing patches Parietal cell 2 times, add the appropriate trypsin digestion cell containing EDTA.Incubation at room temperature to gently piping and druming can blow and beat attached cell When getting off, pancreatin cell dissociation buffer is exhausted as far as possible.The original cell culture fluid collected is added, slightly mixes, is transferred to 5mL centrifuge tubes Interior, 1000g is centrifuged 5 minutes, abandons supernatant, collects cell, cell is gently resuspended with PBS and counts.The cell of 50,000 resuspensions is taken, 1000g is centrifuged 5 minutes, abandons supernatant, adds 100 μ LAimexinV-FITC combination liquid and cell is gently resuspended.Add 2mLAnnexinV- FITC, gently mix and hook.Room temperature lucifuge brightness is educated 10 minutes.1000g is centrifuged 5 minutes, abandons supernatant, is added 2 μ L iodate third and is instigateed dyeing liquor, Gently mix and hook, ice bath avoid light place 15min.Flow cytomery is carried out immediately, and AmexinV-FITC is green fluorescence, and PI is Red fluorescence.AnnexinV-FITC+PI+ and AnnexinV-FITC+PI- is recorded as apoptotic cell, counts 10000 every time Cell, calculate each group apoptosis rate.

4th, statistical procedures

Statistical analysis is carried out using SPSS 16.0, the result that all data represent 3 repetition experiments is tested, with equal Number scholar standard deviation represents.Compare the variance analysis using Factorial Design data between the group of survival rate, compare between the group of apoptosis rate Compared with using one-way analysis of variance, inspection level a=0.05, P<0.05 is to have significant difference, P<0.01 is that pole has significantly Sex differernce.

3rd, result and conclusion

1st, influence of the compound (I) to U251 cells survival rates

Compound (I) suppresses U251 cell survivals growth CCK-8 experimental results and shown, various concentrations compound (I) effect After U251 cells, as activity increases, action time is lengthened, and U251 cells survival rates are decreased obviously, and variance analysis is shown Main effect F=945.8 (the P of compound (I) concentration factor<0.01)；Main effect F=302.67 (the P of action time factor< 0.01), the reciprocation F=45.7 (P of concentration, time factor<0.01).With reference to table 1, it is possible to find compound (I) is thin to U251 The inhibitory action of born of the same parents' survival rate is in time, dose dependent, i.e. compound (I) concentration is higher, and action time is longer, and it suppresses to make With stronger.24h, 48h, 72h IC are acted on by the way that compound (I) is calculated₅₀Value is respectively 3.67mmol/L, 1.37mmol/ L、1.09mmol/L。

2nd, compound (I) dies morphologic influence to U251 cell pillboxes

Normal group and compound (I) treatment group cell dye through Hochest33258, and fluorescence microscopy Microscopic observation is visible right According to a group karyomorphism rule, and it is in light blue uniform coloring；And after 2mmol/L compounds (I) are handled 48 hours, with compareing Group is compared, and different degrees of pyknosis occurs in nucleus, and color is more bright, and the dense dye of stained dense, karyorrhexis is obvious, and visible allusion quotation The apoptotic body of type.The change prompting compound (I) of above Cell Image Analyzer has induction U251 cells apoptosis.

3rd, influence of the compound (I) to U251 apoptosis rates

Control group and each treatment group the row Flow cytometry after Annexin V-FITC/PI dyeing, as a result show：It is right 48 hours group natural death of cerebral cells rates are handled according to group, 1mmol/L compounds (I), 2mmol/L compounds (I) and 4mol/L compounds (I) Respectively 3.30 ± 0.98,14.97 ± 2.67,27.53 ± 5.51 and 55.90 ± 5.1%.Compared with control group, each treatment group The apoptosis rate of cell dramatically increases (P<0.01).As a result Fig. 3 is seen.

Conclusion：Compound (I) is played to human brain glioblastoma mother cell by suppressing cell propagation and inducing cell apoptosis The inhibitory action of knurl U251 cells.These clinical drug research for being found to be compound (I) treatment glioma provide foundation.

Table 1 various concentrations compound (I) processing acts on U251 cells survivals rate (mean ± standard deviation) after 24~72h

Embodiment 3

The preparation of tablet：Compound (I) is first made by the method for embodiment 1, and utilizes organic acid such as tartaric acid or lemon Acid or formic acid or ethanedioic acid etc., inorganic acid salt as made of hydrochloric acid or sulfuric acid or phosphoric acid, it is 1 by itself and excipient weight ratio:9 Ratio adds excipient, pelletizing press sheet.

Embodiment 4

The preparation of oral liquid：Compound (I) is first made by the method for embodiment 1, and utilizes organic acid such as tartaric acid or lemon Lemon acid or formic acid or ethanedioic acid etc., inorganic acid salt as made of hydrochloric acid or sulfuric acid or phosphoric acid, routinely oral liquid preparation method mouth is made Take liquid.

Embodiment 5

The preparation of capsule or granule：Compound (I) is first made by the method for embodiment 1, and utilizes organic acid such as wine Stone acid or citric acid or formic acid or ethanedioic acid etc., the inorganic acid salt as made of hydrochloric acid or sulfuric acid or phosphoric acid, by itself and excipient weight Amount is than being 1:9 ratio adds excipient, and capsule or granule is made.

Embodiment 6

The preparation of parenteral solution：Compound (I) is first made by the method for embodiment 1, and utilizes organic acid such as tartaric acid or lemon Lemon acid or formic acid or ethanedioic acid etc., inorganic acid salt as made of hydrochloric acid or sulfuric acid or phosphoric acid, routinely plus water for injection, refined filtration, Parenteral solution is made in embedding sterilizing.

Embodiment 7

The preparation of aseptic powder injection：By the method for embodiment 1 first be made compound (I), and using organic acid for example tartaric acid or Citric acid or formic acid or ethanedioic acid etc., the inorganic acid salt as made of hydrochloric acid or sulfuric acid or phosphoric acid, are dissolved in sterile water for injection In, stirring makes molten, is filtered with sterile suction funnel, then sterile refined filtration is sub-packed in ampoule, sterile sealing after frozen drying Obtain powder-injection.

The effect of above-described embodiment indicates that the essentiality content of the present invention, but the protection of the present invention is not limited with this Scope.It will be understood by those within the art that technical scheme can be modified or equivalent substitution, Without departing from the essence and protection domain of technical solution of the present invention.

Claims (7)

1. the compound (I) with following structural formula,

2. the preparation method of the compound (I) described in claim 1, it is characterised in that include following operating procedure：(a) by mountain core The dry mature kernal of peach crushes, and is extracted with 75~85% alcohol heat reflux, merges extract solution, is concentrated into no alcohol taste, uses successively Petroleum ether, ethyl acetate and water saturated extracting n-butyl alcohol, respectively obtain petroleum ether extract, acetic acid ethyl ester extract and positive fourth Alcohol extract；(b) acetic acid ethyl ester extract is cleaned with macroreticular resin in step (a), first with 15% ethanol elution, 8 column volumes, Again with 75% ethanol elution, 10 column volumes, 75% ethanol eluate is collected, be concentrated under reduced pressure to obtain 75% ethanol elution thing medicinal extract； (c) 75% ethanol elution thing medicinal extract is separated with purification on normal-phase silica gel in step (b), is successively 85 with volume ratio:1、45:1、20:1、10: 1 and 1:1 methylene chloride-methanol gradient elution obtains 5 components；(d) component 3 is further divided with purification on normal-phase silica gel in step (c) From, successively with volume ratio be 25:1、20:1 and 10:1 methylene chloride-methanol gradient elution obtains 3 components；(e) step (d) The reverse phase silica gel that middle component 2 is bonded with octadecylsilane separates, isocratic with the methanol aqueous solution that concentration expressed in percentage by volume is 75% Elution, collects 8~10 column volume eluents, and eluent is concentrated under reduced pressure to give pure compound (I).

3. the preparation method of compound (I) according to claim 2, it is characterised in that：In step (a), with 80% ethanol Circumfluence distillation, merge extract solution.

4. the preparation method of compound (I) according to claim 2, it is characterised in that：The macroreticular resin is AB-8 types Macroporous absorbent resin.

A kind of 5. pharmaceutical composition, it is characterised in that：Compound (I) described in claim 1 wherein containing therapeutically effective amount And pharmaceutically acceptable carrier.

6. application of the compound (I) in the medicine for preparing treatment glioma described in claim 1.

7. application of the pharmaceutical composition in the medicine for preparing treatment glioma described in claim 5.