CN106008427A - Novel sesquiterpene lactones compound, and preparation method and medical application thereof - Google Patents

Abstract

The invention discloses a novel sesquiterpene lactones compound, and a preparation method and medical application thereof, and belongs to the technical field of medicine. The novel sesquiterpene lactones compound is reported for the first time, has a novel structure and can be obtained by extracting, separating and purifying dry rhizomes of valeriana jatamansi Jone. An in-vitro test shows that the novel sesquiterpene lactones compound can be used for inhibiting the proliferation of gastric cancer cells and promoting the apoptosis of cells, the growth inhibition function on the gastric cancer cells is in an enhancing trend along with the increment of the concentration, and the novel sesquiterpene lactones compound can be developed into medicine for treating gastric cancer.

Description

A kind of new sesquiterpene lactones compounds and preparation method thereof and medical usage

Technical field

The invention belongs to technical field of pharmaceuticals, be specifically related to a kind of sesquialter of isolated from the dry rhizome of Rhizoma valerianae latifoliae Terpene lactones compound, containing its pharmaceutical composition and its preparation method and application.

Background technology

Rhizoma valerianae latifoliae (Valeriana wallichii DC. or Valerianajatamansi Jone) has another name called Herba Asari, soil Herba Asari, Indian Rhizoma et radix valerianae etc., be Herba Patriniae section valerian, and plant high 20～70cm, rhizome is the thickest, block column, saves close, has dense Strong fragrance.Rhizoma valerianae latifoliae is herbaceos perennial, and happiness is longer than in the meadow, mountain top of below height above sea level 2500m, spinney or small stream limit, China is distributed mainly on the ground such as Shaanxi, Henan, Hubei, Sichuan, Guizhou, Yunnan.Rhizoma valerianae latifoliae rhizome has abundant medical value, Chinese Traditional Medicine thinks that it has the effects such as regulating QI to relieve pain, antiinflammatory antidiarrheal, expelling wind and removing dampness, tranquillizing and allaying excitement, can be used for treating gastral cavity abdomen The diseases such as distending pain, dyspepsia, diarrhoea, dysentery, rheumatic arthralgia, snake venom, insomnia, obesity.

Modern study shows, the main chemical compositions of Rhizoma valerianae latifoliae has volatile oil, iridoids and flavonoid etc..Rhizoma valerianae latifoliae Medical value greatly owing to the abundant volatile oil contained by its root and stem, its chemical composition is more complicated, mainly wraps Include monoterpene alkenes, sesquiterpene and containing oxygen derivative thereof etc..

Modern study shows that Rhizoma valerianae latifoliae has antitumor action, calmness, analgesia, angst resistance effect and anti-rotavirus Deng effect.

Summary of the invention

It is an object of the invention to provide a kind of a kind of Sesquiterpene lactones of isolated from the dry rhizome of Rhizoma valerianae latifoliae Compound, containing its pharmaceutical composition and its preparation method and application.

The above-mentioned purpose of the present invention is achieved by techniques below scheme:

There is the compound (I) of following structural formula:

The preparation method of described compound (I), comprises following operating procedure: the dry rhizome of Rhizoma valerianae latifoliae is pulverized by (a), With 70～80% alcohol heat reflux extract, united extraction liquid, be concentrated into without alcohol taste, full with petroleum ether, ethyl acetate and water successively The n-butanol extraction of sum, respectively obtains petroleum ether extract, acetic acid ethyl ester extract and n-butyl alcohol extract；In (b) step (a) Acetic acid ethyl ester extract macroporous resin remove impurity, first with 6 column volumes of 5% ethanol elution, then with 8 cylinders of 70% ethanol elution Long-pending, collect 70% eluent, concentrating under reduced pressure obtains 70% ethanol elution concentrate；70% ethanol elution concentrate in (c) step (b) Separate by purification on normal-phase silica gel, obtain 4 with the methylene chloride-methanol gradient elution that volume ratio is 50:1,30:1,15:1 and 5:1 successively Individual component；D in () step (c), component 4 separates further by purification on normal-phase silica gel, be the dichloro of 8:1,5:1 and 2:1 by volume ratio successively Methane-methanol gradient elution obtains 3 components；E reverse phase silica gel that in () step (d), component 2 is bonded by octadecylsilane divides From, with the methanol aqueous solution isocratic elution that concentration expressed in percentage by volume is 70%, collect 8～12 column volume eluents, eluent subtracts Pressure is concentrated to give pure compound (I).

Further, in step (a), extract with 75% alcohol heat reflux, united extraction liquid.

Further, described macroporous resin is AB-8 type macroporous adsorbent resin.

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

The application in the medicine of preparation treatment gastric cancer of the described compound (I).

The application in the medicine of preparation treatment gastric cancer of the described pharmaceutical composition.

When the compounds of this invention is used as medicine, can directly use, or use with the form of pharmaceutical composition.

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

Described pharmaceutically suitable carrier or excipient are that one or more are selected from solid, semisolid and liquid diluent, filler And pharmaceutical preparation adjuvant.The pharmaceutical composition of the present invention is used with the form of per weight dose.Medicine of the present invention can It is applied to need the patient for the treatment of by oral or injection form.During for being administered orally, tablet, slow releasing tablet, control can be made into Release sheet, capsule, drop pill, micropill, suspensoid, Emulsion, powder or granule, oral liquid etc.；During for injecting, can be made into sterilizing Aqueous or oily solution, aseptic powder injection, liposome or Emulsion etc..

Accompanying drawing explanation

Fig. 1 is compound (I) structural formula；

Fig. 2 is that compound (I) calculates ECD and experiment ECD figure.

Detailed description of the invention

Further illustrate the essentiality content of the present invention below in conjunction with embodiment, but do not limit the present invention with this and protect model Enclose.Although the present invention being explained in detail with reference to preferred embodiment, it will be understood by those within the art that, can be right Technical scheme is modified or equivalent, without deviating from the spirit and scope of technical solution of the present invention.

Main material, reagent source and instrument type:

Ethanol, petroleum ether, ethyl acetate, n-butyl alcohol, dichloromethane are analytical pure, limited purchased from Shanghai Ling Feng chemical reagent Company, methanol, analytical pure, purchased from Jiangsu Han Bang chemical reagent company limited.

Embodiment 1: compound (I) separates preparation and structural identification

A the dry rhizome (8kg) of Rhizoma valerianae latifoliae is pulverized by (), extract (30L × 3 time) with 75% alcohol heat reflux, and merging carries Take liquid, be concentrated into without alcohol taste (6L), successively with petroleum ether (6L × 3 time), ethyl acetate (6L × 3 time) and water saturated n-butyl alcohol (6L × 3 time) extract, and respectively obtain petroleum ether extract, acetic acid ethyl ester extract (375g) and n-butyl alcohol extract；(b) step A acetic acid ethyl ester extract AB-8 macroporous resin remove impurity in (), first with 6 column volumes of 5% ethanol elution, then washes with 70% ethanol De-8 column volumes, collect 70% eluent, and concentrating under reduced pressure obtains 70% ethanol elution concentrate (129g)；In (c) step (b) 70% ethanol elution concentrate purification on normal-phase silica gel separates, and is 50:1 (8 column volumes), 30:1 (8 cylinders by volume ratio successively Long-pending), the methylene chloride-methanol gradient elution of 15:1 (8 column volumes) and 5:1 (10 column volumes) obtain 4 components；(d) step Suddenly in (c), component 4 (27g) separates further by purification on normal-phase silica gel, is 8:1 (8 column volumes), 5:1 (10 posts by volume ratio successively Volume) and the methylene chloride-methanol gradient elution of 2:1 (8 column volumes) obtain 3 components；Component 2 in (e) step (d) (15g) separate with the reverse phase silica gel of octadecylsilane bonding, wash with the methanol aqueous solution that concentration expressed in percentage by volume is 70% is isocratic De-, collect 8～12 column volume eluents, eluent is concentrated under reduced pressure to give pure compound (I) (138mg).

Structural identification: white crystal；HR-ESIMS shows [M+Na]⁺For m/z 469.1804, can obtain in conjunction with nuclear-magnetism feature Molecular formula is C₂₄H₃₀O₈, degree of unsaturation is 10.Hydrogen nuclear magnetic resonance modal data δ_H(ppm, DMSO-d₆, 400MHz): H-1 (4.71, Dd, J=9.8,6.8), and H-2 (2.37, m), H-2 (1.82, m), H-3 (5.25, br, s), H-5 (2.40, br, d, J=12.5), H-6 (3.89, t, J=13.4), H-7 (2.73, tt, J=10.9,2.9), H-8 (5.14, td, J=10.8,4.5), H-9 (1.06, br, t, J=9.2), H-9 (2.04, dd, J=12.6,4.5), H-13 (6.01, d, J=2.8), H-13 (5.35, d, J =2.8), and H-14 (0.96, s), H-15 (1.72, br, s), H-18 (5.47, q, J=6.2), H-19 (6.62, br, s), H-19 (6.04, br, s), H-20 (1.46, d, J=6.2), H-22 (2.11, s), H-24 (2.01, s)；Carbon-13 nmr spectra data δ_C (ppm, DMSO-d₆, 100MHz): 75.5 (CH, 1-C), 28.6 (CH₂, 2-C), 121.1 (CH, 3-C), 131.5 (C, 4-C), 49.3 (CH, 5-C), 77.8 (CH, 6-C), 52.7 (CH, 7-C), 70.3 (CH, 8-C), 39.4 (CH₂, 9-C), 38.4 (C, 10- C), 135.5 (C, 11-C), 168.6 (C, 12-C), 118.4 (CH₂, 13-C), 13.1 (CH₃, 14-C), 22.7 (CH₃, 15-C), 165.8 (C, 16-C), 139.6 (C, 17-C), 65.2 (CH, 18-C), 126.7 (CH₂, 19-C), 19.1 (CH₃, 20-C), 169.0 (C, 21-C), 20.4 (CH₃, 22-C), 169.8 (C, 23-C), 20.7 (CH₃, 24-C)；Carbon atom labelling sees Fig. 1.IR spectrum Show that this compound contains double bond (1643cm^-1) and ester group (1782 and 1736cm^-1) functional group.¹H and¹³C H NMR spectroscopy shows four Individual oxygen-containing methine signals [δ H4.71 (1H, dd, J=9.8,6.8Hz, H-1), 3.89 (1H, t, J=13.4Hz, H-6), 5.14 (1H, td, J=10.8,4.5HZ, H-8) and 5.47 (1H, q, J=6.2Hz, H-18)], a bimodal methine resonance signal [δ H2.40 (1H, br, d, J=12.5Hz, H-5)], an olefinic methine signals [δ H5.25 (1H, br, s, H-3)], two rings Outer methene proton signal [δ H6.01 (1H, d, J=2.8Hz, H-13a) and 5.35 (1H, d, J=2.8Hz, H-13b), 6.62 (1H, br, s, H-19a) and 6.04 (1H, br, s, H-19b)], three methyl signals [δ H0.96 (3H, s, H-14), 1.72 (3H, Br, s, H-15) and 1.46 (3H, d, J=6.2Hz, H-20)], two acetoxyl groups [δ H2.11 (3H, s, H-22), 2.01 (3H, S, H-24)；δ C169.0 (C-21), 169.8 (C-23)].In HMBC spectrum, H-1 Yu C-23 (δ C169.8), Me-24 Yu C-1 (δ And the dependency of H-18 Yu C-21 (δ C169.0) shows that C-1 and C-18 position is respectively connected with an acetoxyl group C75.5),.Additionally, Owing to H-18 is quartet, Me-20 is doublet, shows that C-18 position is also connected with a methyl.In HMBC spectrum, H-8 Yu C-16 (δ C165.8) dependency shows that C-8 position is connected with an ester group.H in being composed by HMBC₂The dependency of-19 and C-17 (δ C139.6) Understand and there is double bond between C-17 and C-19.H-8 and H-6 in NOESY spectrum, and the dependency of H-6 Yu Me-14 shows these matter Son is beta comfiguration.Comprehensive hydrogen spectrum, carbon spectrum, HMBC spectrum and NOESY spectrum, and document is about correlation type nuclear magnetic data, can be the most true This compound fixed is as it is shown in figure 1, spatial configuration is determined by ECD test further, and theoretical value is basically identical with experiment value (to be schemed 2)。

Embodiment 2: compound (I) pharmacological action is tested

One, material and instrument

Human stomach cancer cell line SGC-7901 is purchased from Sai Er Reagent Company.Compound (I) is made by oneself, and HPLC normalization purity is more than 98%.PDTC, trypsin, MTT, dimethyl sulfoxide (DMSO), agarose, glacial acetic acid are purchased from Sigma Co., USA.RPMI- 1640 culture medium, PBS phosphate buffered solution are purchased from GIBCO company of the U.S..Hyclone is purchased from Shandong Yin Xiang great achievement company.Platform Expect indigo plant (Beijing Bioisystech Co., Ltd of Zhong Shan Golden Bridge), TUNEL test kit (Kai Ji biotechnology Development Co., Ltd), DAB Colour reagent box (Beijing biotech firm of Zhong Shan Golden Bridge), formaldehyde (Fisher company of the U.S.), (sky, Tianjin is the most smart for catalase Refinement work development centre).

WD-9403C uv analyzer (Biometra company of Germany), grads PCR instrument (Biometra company of Germany), gel Image analysis system (Shanghai Tian Neng company), electrophresis apparatus (Beijing 6 1), electronic balance (the limited public affairs of Shanghai precision instrumentation Department), RKI-1002 type CO2 gas incubator (Ikemoto company of Japan), (it is limited that Suzhou purifies experimental facilities to superclean bench Company), supercentrifuge (Beijing medical apparatus and instruments factory), low speed centrifuge (Beijing medical apparatus and instruments factory), Wilovert S type is inverted Microscope (Olympus company of Japan), vertical pressure steam sterilizer (Shanghai Bo Xun Industrial Co., Ltd.), cell counting count board (Shanghai precision instrument company), ultra-pure water demineralizer (Britain PURELAB ulTRA GENETIC).

Two, test method

1, the cultivation of stomach cancer cell SGC-7901

1.1 cell recovery

(1) from liquid nitrogen container, take out cell cryopreservation tube, be immediately placed in 37 DEG C～42 DEG C, in 75% ethanol, then move to same In warm water bath；(2) rock l～3min cryopreservation tube gently, make frozen stock solution melt, move to rapidly in super-clean bench；(3) will be containing thin The frozen stock solution of born of the same parents sucks sterile centrifugation tube, adds appropriate RPIM-1640 culture medium；(4) low speed centrifuge is put into after piping and druming mixing, 800rpm/min is centrifuged 3 minutes, removes supernatant；(5) add appropriate culture medium and blow even, cell suspension is inoculated into according to concentration In 10mL culture dish；(6) be placed in containing 5% carbon dioxide, 37 DEG C of saturated humidities incubator in cultivate.

1.2 passage

(1) during the cell attachment in culture dish about 80%, with the culture medium piping and druming that pipette, extract is old in super-clean bench Cell in culture dish for several times, to blow dead cell off；(2) suck old culture medium with pipet, add appropriate 0.25% pancreatin and disappear Change cell, be placed in 30 DEG C of incubators digestion；(3) after about 3min, culture dish is placed under inverted microscope observation, treats cell week Limit is shinny, retraction then illustrates after becoming round that cell departs from from wall；(4) in super-clean bench, pancreatin is sucked rapidly.Add appropriate Culture medium blows and beats cell repeatedly, makes cell completely disengage from culture dish；(5) cell suspension is seeded to respectively different trainings by concentration Support in ware, supply culture medium；(6) it is replaced in containing 5%CO₂, 37 DEG C of saturated humidities incubator in cultivate.

2, cell counting

(1) getting out cell counting count board and coverslip, both is clean by alcohol wipe, covers coverslip at counting chamber On；(2) after ethanol volatilizees, the appropriate cell suspension of sucking-off, dropping, at coverslip edge, makes suspension be full of coverslip and counting chamber Between, it is careful not to overflow coverslip and the glass guide channel of both sides, counts after standing；(3) 4 big lattice are found under the microscope, often Individual big lattice are divided into again 16 little lattice, and the cell number in 4 big lattice of counting, averages respectively.On the left of line ball cytometer and top , disregard right side and lower section；(4) average cell number × 10000 of cell concentration (individual/mL)=each grid；(5) by cell Suspension is diluted to test desired concn.

3, cell viability measures

(1) SGC-7901 stomach cancer cell suspension 0.9mL to be determined is taken；(2) in cell suspension, trypan blue piping and druming is added Mixing；(3) use cell counting count board blind at least 200 cells of counting, observe under inverted microscope；(4) Microscopic observation cell Staining conditions, intracellular dyes light blue person for dead cell, and the person of being unstained is living cells, with hundred of total cellular score shared by living cells The vigor (%) of proportion by subtraction reflection cell.

4, MTT detects inhibitory rate of cell growth

Experiment packet: 1. negative control group；2. compound (I) group 1, compound (I) (50 μm ol/L)；3. compound (I) Group 2, compound (I) (100 μm ol/L)；4. PDTC group 1, PDTC (50 μm ol/L)；5. PDTC group 2, PDTC (100 μm ol/L)； 6. drug combination group 1, compound (I) (25 μm ol/L)+PDTC (25 μm ol/L)；7. drug combination group 2, compound (I) (50 μ mol/L)+PDTC(50μmol/L)。

(1) taking the logarithm the SGC-7901 cell of trophophase, count with cell counting count board, regulation cell concentration is 5 × l0⁵/ mL；(2) sample injector is used to be inoculated in 96 orifice plates with every hole 100 μ L.Note only being inoculated into during inoculation 60 holes of centre, periphery 36 Hole is filled with PBS, simultaneously 3 96 orifice plates of paving；(3) 96 orifice plates completed are placed in 37 DEG C, 5%CO₂In cell culture incubator, treat Take out after 24h cell attachment；(4) 96 orifice plates are divided into compound (I) group, (0,50,100 μm ol/L), PDTC group (0,50,100 μm ol/L), group (0/0,25/25,50/50 μm ol/L) combined by two medicines, sets up the most celliferous blank group (only to add training simultaneously Support base) give different disposal respectively；(5) each concentration of each medicine sets 5 multiple holes, cultivates 24,48 and 72h respectively；(6) exist respectively Specific end time point takes out 96 orifice plates, and every hole adds MTT (5g/L) solution 20 μ L, puts back to incubator and continues to cultivate 4h, eventually Only cultivate.(7) carefully sucking supernatant in hole, every hole adds 150 μ L DMSO, plate shaker vibration 10min makes crystallization the most molten Solving, basis of microscopic observation granule disappears.(8) at microplate reader 490nm wavelength, optical density (OD) value in each hole is measured, when calculating each Between point inhibitory rate of cell growth.Suppression ratio=[1-(dosing group OD value-blank group OD value)/(negative control group OD value- Blank group OD value)] × 100%.

5, TUNEL method detection natural death of cerebral cells index

The preparation of 5.1 cell climbing sheets

(1) process of coverslip: coverslip is placed in soaked overnight in concentrated sulphuric acid, first rinses for several times with tap water next day, It is placed in dehydrated alcohol immersion 4h again, then rinses well with deionized water, be placed on for dry case is dried the sterilization of laggard horizontal high voltage, Taking-up is directly placed in super-clean bench standby.In 6 orifice plates are placed in super-clean bench, ultraviolet irradiates 30min；(2) coverslip is placed: six Every hole of orifice plate prepares to put the position of coverslip instill after a small amount of culture medium and place coverslip again so that coverslip is examined with orifice plate The tension force of culture medium is bonded together, and when preventing from adding cell suspension, coverslip hikes up, and causes double-layer cell adherent；(3) take the logarithm The SGC-7901 cell of trophophase, cell suspension is blown and beaten in digestion, is l × 10 with cell counting count board adjustment cell concentration⁶/mL。 (4) it is separately added into 1mL cell suspension with sample injector in the every hole being placed with coverslip, spreads 3 plates simultaneously, be placed in 37 DEG C, 5%CO₂Training Support cultivation 24h in case and treat cell attachment；In super-clean bench, 6 orifice plates are divided into after (5) 24 hour cells are adherent negative control group and Experimental group gives different disposal respectively, and negative control group adds 1mL culture medium, experimental group be further divided into compound (I) group, PDTC group and 3 subgroups of drug combination group, every hole adds 1mL medicine.Compound (I) organizes final concentration of 100 μm ol/L, and PDTC group is final concentration of 100 μm ol/L, the final concentration of two kinds of medicines of group combined by two medicines is respectively 50 μm ol/L.Often group sets 2 multiple holes.(6) 37 DEG C it are placed in, 5%CO₂Cell culture incubator in continue cultivate.

5.2TUNEL method operating procedure

(1) cell climbing sheet dosing takes out 6 orifice plates when cultivating 24h, carries out following steps successively according to TUNEL description；(2) Careful suction abandons the supernatant in every hole；(3) each hole adds PBS (4 DEG C) 2mL, and plate shaker rinses 5min × 3 time；(4) every hole Adding 1mL pre-cooling cell fixative, 30min fixed by 4 DEG C of refrigerators.(5) fixative is abandoned in suction, and each hole adds PBS (4 DEG C) 2mL, flat Plate shaking table rinses 5min × 3 time.(6) immersing in confining liquid, room temperature (15 DEG C～25 DEG C) closes 10min.(7) each hole adds PBS (4 DEG C) 2mL, plate shaker rinses 5min × 3 time.Blot with absorbent paper around sample.(8) each sample drips 50 μ L TdT enzyme reaction solution, 37 DEG C, lucifuge moistening reaction 60min.(9) each hole adds PBS (4 DEG C) 2mL, and plate shaker rinses 5min × 3 times.Blot with absorbent paper around sample.(10) dripping 50 μ L Streptavidin-HRP working solutions, 37 DEG C, lucifuge moistens Reaction 30min.(11) each hole adds PBS (4 DEG C) 2mL, and plate shaker rinses 5min × 3 time.(12) 100 μ L DAB works are dripped Make liquid, color development at room temperature reaction 10min.(13) each hole adds PBS (4 DEG C) 2mL, and plate shaker rinses 5min × 3 time.(14) light Basis of microscopic observation is taken pictures: randomly selects 10 high power (× 100) visuals field, each visual field 100 cells of counting, calculates tune and die The meansigma methods of index.Apoptotic index (AI)=(positive cell number/total cell number × 100%).The nucleus of positive cell is palm fibre Brown.

6, statistical analysis

Use SPSS 11.5 be analyzed, meet JH state distribution measurement data to represent, compare between group and use Oneway- ANOVA method is analyzed, and compares employing LSD-t method two-by-two, and P < 0.05 is that difference is statistically significant.

Three, result and conclusion

1, MTT detects the medicine growth inhibition ratio to SGC-7901 stomach cancer cell

Repeating MTT through 3 times, testing result shows, after single medicine acts on stomach cancer cell 72h, and the medicine of 100 μm ol/L Group is the highest to the growth inhibition ratio of stomach cancer cell compared with the medicine group of 50 μm ol/L, difference statistically significant (P < 0.05).50μ The growth inhibited effect of stomach cancer cell is anticipated by the compound (I) of mol/L with drug combination 25/25 μm ol/L group no statistical difference Justice (P > 0.05).The PDTC of 50 μm ol/L to the growth inhibited effect of stomach cancer cell compared with drug combination 25/25 μm ol/L group, Difference statistically significant (P < 0.05).Drug combination 50/50 μm ol/L group is higher than each to the growth inhibition ratio of stomach cancer cell High concentration list medicine group (the 100 μm ol/L) growth inhibition ratio to stomach cancer cell, difference statistically significant (P < 0.001), it is shown in Table 1 (* P < 0.05, * * P < 0.01).

2, each group of apoptotic index of TUNEL method detection

Compound (I), PDTC and drug combination group all have the effect of Developing restraint to stomach cancer cell SGC-7901, each group with Negative control group comparing difference the most statistically significant (P < 0.001), negative control group has the karyon of a small amount of cell to be dyed to palm fibre Brown, each single medicine group all has increase compared with drug combination group apoptotic cell relatively negative control group, drug combination group cell Apoptotic index is the highest, more notable to the inhibition of stomach cancer cell.It is shown in Table 2 (* * P < 0.01).

Conclusion, PDTC and compound (I) two medicine act solely on stomach cancer cell and all can promote with the propagation of anticancer The apoptosis of cell, along with the increase of concentration, to the Developing restraint effect of stomach cancer cell in strengthening trend, two kinds of Drug combinations The apoptosis effect of rear promotion stomach cancer cell is more notable, and the growth inhibition ratio of cell and apoptotic index the most independent medication group raise. Research finds that PDTC and (I) two kind of Drug combination of compound are more notable to the inhibitory action of stomach cancer cell, and two kinds of medicines Being the chemotherapeutics of non-traditional meaning, the infringement to body is little, and avoids resistance to classic chemotherapy medicine of tumor cell The property of medicine, can be that the clinical treatment of gastric cancer from now on provides reference.

Table 1 each medicine group different time points to the growth inhibition ratio of stomach cancer cell (n=5,)

Group	n	24h	48h	72h
					Compound (I) 50 μm ol/L (1)	5	14.87±5.19	16.29±4.53	55.70±6.67
Compound (I) 50 μm ol/L (2)	5	31.00±6.36	59.42±7.74	74.32±5.84
					PDTC50μmol/L(3)	5	24.87±9.04	36.83±5.21	40.58±7.15
PDTC100μmol/L(4)	5	42.86±6.28	44.21±8.44	50.31±4.63
					Drug combination 25/25 μm ol/L (5)	5	36.43±8.13	38.76±10.87	58.55±9.53
Drug combination 50/50 μm ol/L (6)	5	49.38±2.24	74.56±10.59	96.12±2.25
					F		17.918**	29.646**	47.625**
P(1):(2)		0.001	<0.001	<0.001
					(3):(4)		<0.001	0.169	0.025
(5):(6)		0.005	<0.001	<0.001
					(1):(5)		<0.001	<0.001	0.49
(3):(5)		0.01	0.713	<0.001
					(2):(6)		<0.001	0.008	<0.001
(4):(6)		0.13	<0.001	<0.001

After table 2 medicine effect 24h each group cell apoptotic index (n=3,)

Embodiment 3

The preparation of tablet: first prepare compound (I) by embodiment 1 method, and utilize organic acid such as tartaric acid or Fructus Citri Limoniae The salt that acid or formic acid or ethanedioic acid etc., mineral acid example hydrochloric acid or sulphuric acid or phosphoric acid are made, by its with excipient weight ratio for 1:7's Ratio adds excipient, pelletizing press sheet.

Embodiment 4

The preparation of oral liquid: first prepare compound (I) by embodiment 1 method, and utilize organic acid such as tartaric acid or lemon The salt that lemon acid or formic acid or ethanedioic acid etc., mineral acid example hydrochloric acid or sulphuric acid or phosphoric acid are made, oral liquid preparation method makes mouth routinely Take liquid.

Embodiment 5

Capsule or the preparation of granule: first prepare compound (I) by embodiment 1 method, and utilize organic acid such as wine The salt that stone acid or citric acid or formic acid or ethanedioic acid etc., mineral acid example hydrochloric acid or sulphuric acid or phosphoric acid are made, by itself and excipient weight Amount adds excipient than the ratio for 1:9, makes capsule or granule.

Embodiment 6

The preparation of injection: first prepare compound (I) by embodiment 1 method, and utilize organic acid such as tartaric acid or lemon The salt that lemon acid or formic acid or ethanedioic acid etc., mineral acid example hydrochloric acid or sulphuric acid or phosphoric acid are made, injects routinely and uses water, fine straining, Injection is made in embedding sterilizing.

Embodiment 7

The preparation of aseptic powder injection: first prepare compound (I) by embodiment 1 method, and utilize organic acid such as tartaric acid or The salt that citric acid or formic acid or ethanedioic acid etc., mineral acid example hydrochloric acid or sulphuric acid or phosphoric acid are made, is dissolved in sterile water for injection In, stirring makes molten, filters with aseptic suction funnel, more aseptic fine straining, is sub-packed in ampoule, aseptic sealing by fusing after frozen drying Obtain injectable powder.

The effect of above-described embodiment indicates that the essentiality content of the present invention, but does not limit the protection of the present invention with this Scope.It will be understood by those within the art that, technical scheme can be modified or equivalent, Essence and protection domain without deviating from technical solution of the present invention.

Claims (7)

1. there is the compound (I) of following structural formula:

。

2. the preparation method of the compound (I) described in claim 1, it is characterised in that comprise following operating procedure: (a) is by Aranea Fragrant dry rhizome is pulverized, and extracts with 70 ~ 80% alcohol heat reflux, united extraction liquid, is concentrated into without alcohol taste, use successively petroleum ether, Ethyl acetate and water saturated n-butanol extraction, respectively obtain petroleum ether extract, acetic acid ethyl ester extract and n-butanol extraction Thing；B in () step (a), n-butyl alcohol takes thing macroporous resin remove impurity, first with 6 column volumes of 5% ethanol elution, then wash with 70% ethanol De-8 column volumes, collect 70% eluent, and concentrating under reduced pressure obtains 70% ethanol elution concentrate；70% ethanol elution in (c) step (b) Concentrate purification on normal-phase silica gel separates, successively with the methylene chloride-methanol gradient elution that volume ratio is 50:1,30:1,15:1 and 5:1 Obtain 4 components；D in () step (c), component 4 separates further by purification on normal-phase silica gel, be 8:1,5:1 and 2:1 by volume ratio successively Methylene chloride-methanol gradient elution obtain 3 components；In (e) step (d) component 2 with octadecylsilane be bonded anti-phase Silica gel separates, and with the methanol aqueous solution isocratic elution that concentration expressed in percentage by volume is 70%, collects 8 ~ 12 column volume eluents, eluting Liquid is concentrated under reduced pressure to give pure compound (I).

The preparation method of compound the most according to claim 2 (I), it is characterised in that: in step (a), by 75% ethanol heat Reflux, extract, united extraction liquid.

The preparation method of compound the most according to claim 2 (I), it is characterised in that: described macroporous resin is AB-8 type Macroporous adsorbent resin.

5. a pharmaceutical composition, it is characterised in that: wherein contain the compound (I) described in the claim 1 of therapeutically effective amount With pharmaceutically acceptable carrier.

6. the application in the medicine of preparation treatment gastric cancer of the compound (I) described in claim 1.

7. the application in the medicine of preparation treatment gastric cancer of the pharmaceutical composition described in claim 5.