CN104069273A - A traditional Chinese medicine composition having anticancer effects and a preparing method and uses thereof - Google Patents
A traditional Chinese medicine composition having anticancer effects and a preparing method and uses thereof Download PDFInfo
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Abstract
The invention provides a traditional Chinese medicine composition having anticancer effects and a preparing method and uses thereof. The traditional Chinese medicine composition mainly comprises following raw material medicines: trogopterus dung and cattail pollen. The preparing method includes: weighing the raw material medicines according to a ratio, and extracting with water or an organic solvent mutually soluble with water; or weighing the raw material medicines according to the ratio and preparing by adoption of a water extraction and alcohol precipitation method, wherein the alcohol precipitation concentration is 50-90%. The uses of the composition relate to applications of the composition in preparation of medicines treating cancer especially medicines treating cervical cancer. The composition comprises the trogopterus dung and the cattail pollen in a weight ratio of 1:0.2-4. By compatibility of the trogopterus dung and the cattail pollen, the composition has coordinate repression effects for Hela cells and has good anticancer effects.
Description
Technical field
The present invention relates to a kind of Chinese medicine composition and its production and use, be specifically related to a kind of Chinese medicine composition with antitumaous effect and its production and use, belong to Chinese medicine technical field.
Background technology
Cancer is the major disease that jeopardizes human health, and its fatality rate is the trend rising year by year, and its generation and development are the complex lesions processes of multifactor participation, polygenes change and Multi stage development.Treatment for cancer at present adopts operative treatment in conjunction with Radiotherapy chemotherapy method mostly, but the method is large to patient's wound, dangerous high, and somewhat expensive, but curative effect is unsatisfactory.The weapon that Chinese medicine is prevented and cured diseases as China's tradition, is accompanied by recent years progressively deep research and discovery, in treatment of cancer, more and more comes into one's own.Research shows, in treatment and prevention of tumour process, Chinese medicine is mainly shift and the aspects such as attenuation synergistic of chemotherapy are played a role from improving body's immunity, inducing apoptosis of tumour cell, inhibition tumor cell.
Chinese medicine composition compatibility of the present invention is reasonable, can regulate negative and positive of qi and blood and function, and protection hemopoietic function of bone marrow, can start self-defense system again, improves immunity of organisms, very helpful for the auxiliary treatment of tumor.
Summary of the invention
First object of the present invention is to provide a kind of Chinese medicine composition with antitumaous effect.
Second object of the present invention is to provide the preparation method of said composition.
The 3rd object of the present invention is to provide the application of said composition in preparation treatment cancer drug.
The 4th object of the present invention is to provide the application of said composition in preparation treatment cervical cancer medicine.
The object of the invention is to be achieved through the following technical solutions:
A Chinese medicine composition with antitumaous effect, this Chinese medicine composition is comprised of the crude drug of following weight ratio:
Oletum Trogopterori: Pollen Typhae=1:(0.2-4).
The crude drug of described Chinese medicine composition is preferably:
Oletum Trogopterori: Pollen Typhae=1:(0.3-3.5).
The crude drug of described Chinese medicine composition is preferably:
Oletum Trogopterori: Pollen Typhae=1:(0.4-3).
The crude drug of described Chinese medicine composition is preferably:
Oletum Trogopterori: Pollen Typhae=1:(0.5-2.5).
The crude drug of described Chinese medicine composition is preferably:
Oletum Trogopterori: Pollen Typhae=1:(0.5-2.2).
The crude drug of described Chinese medicine composition is preferably:
Oletum Trogopterori: Pollen Typhae=1:(0.5-2).
The crude drug of described Chinese medicine composition is more preferably:
Oletum Trogopterori: Pollen Typhae=1:0.5;
Or, Oletum Trogopterori: Pollen Typhae=1:1;
Or, Oletum Trogopterori: Pollen Typhae=1:2.
Chinese medicine composition of the present invention can be prepared as follows:
Get in proportion crude drug, with water or the organic solvent extraction that dissolves each other with water;
Further, the described organic solvent dissolving each other with water is selected from one or more in methanol, ethanol, acetone; Further be preferably the ethanol of 50-90%; Further be preferably the ethanol of 60-80%; Further be preferably 70% ethanol;
Extracting method used comprises any one mode decocting in extraction, reflux, extract,, immersion extraction, supersound extraction or percolation extraction, or the combination of Different Extraction Method.
Chinese medicine composition of the present invention also can adopt: get in proportion crude drug, adopt decoction and alcohol sedimentation technique preparation; Wherein said alcohol precipitation concentration is 50-90%;
Further, described alcohol precipitation concentration is 60-80%; Further described alcohol precipitation concentration is 70%.
Above-mentioned preparation method Raw medicine also can be purified, refining after extracting, as crossed macroporous resin column; And further preparation process is routinely made the acceptable any conventional dosage form of pharmaceutics, comprises granule, tablet, capsule, drop pill, oral liquid, suspension, emulsion, injection.
For above-mentioned dosage form can be realized, need when these dosage forms of preparation, add the acceptable adjuvant of pharmacy, such as: filler, disintegrating agent, lubricant, suspending agent, binding agent, sweeting agent, correctives, antiseptic, substrate etc.Filler comprises: starch, pregelatinized Starch, lactose, mannitol, chitin, microcrystalline Cellulose, sucrose etc.; Disintegrating agent comprises: starch, pregelatinized Starch, microcrystalline Cellulose, carboxymethyl starch sodium, crospolyvinylpyrrolidone, low-substituted hydroxypropyl cellulose, cross-linking sodium carboxymethyl cellulose etc.; Lubricant comprises: magnesium stearate, sodium lauryl sulphate, Pulvis Talci, silicon dioxide etc.; Suspending agent comprises: polyvinylpyrrolidone, microcrystalline Cellulose, sucrose, agar, hydroxypropyl emthylcellulose etc.; Binding agent comprises: starch slurry, polyvinylpyrrolidone, hydroxypropyl emthylcellulose etc.; Sweeting agent comprises: saccharin sodium, Aspartane, sucrose, cyclamate, enoxolone etc.; Correctives comprises: sweeting agent and various essence; Antiseptic comprises: parabens, benzoic acid, sodium benzoate, sorbic acid and its esters, benzalkonium bromide, acetic acid chloroethene are determined, Folium eucalypti globueli (Eucalyptus globulus Labill.) wet goods; Substrate comprises: PEG6000, PEG4000, insect wax etc.For making above-mentioned dosage form can realize pharmacy of Chinese materia medica, need add acceptable other adjuvant of pharmacy (the green booth < of model < pharmacy of Chinese materia medica > >, the adjuvant that in Shanghai Science Press December in 1997 the 1st edition, each dosage form is recorded) during these dosage forms in preparation.
Chinese medicine composition of the present invention is except the form feeding intake with Oletum Trogopterori, Pollen Typhae crude drug, can also adopt the form feeding intake with Oletum Trogopterori, Pollen Tyjphae extract (effective site), therefore the present invention further discloses a kind of Chinese medicine composition with antitumaous effect:
A Chinese medicine composition with antitumaous effect, this Chinese medicine composition is comprised of the raw material of following weight ratio:
Oletum Trogopterori extract: Pollen Tyjphae extract=1:(0.2-4).
The crude drug of described Chinese medicine composition is preferably:
Oletum Trogopterori extract: Pollen Tyjphae extract=1:(0.3-3.5).
The crude drug of described Chinese medicine composition is preferably:
Oletum Trogopterori extract: Pollen Tyjphae extract=1:(0.4-3).
The crude drug of described Chinese medicine composition is preferably:
Oletum Trogopterori extract: Pollen Tyjphae extract=1:(0.5-2.5).
The crude drug of described Chinese medicine composition is preferably:
Oletum Trogopterori extract: Pollen Tyjphae extract=1:(0.5-2.2).
The crude drug of described Chinese medicine composition is preferably:
Oletum Trogopterori extract: Pollen Tyjphae extract=1:(0.5-2).
The crude drug of described Chinese medicine composition is more preferably:
Oletum Trogopterori extract: Pollen Tyjphae extract=1:0.5;
Or, Oletum Trogopterori extract: Pollen Tyjphae extract=1:1;
Or, Oletum Trogopterori extract: Pollen Tyjphae extract=1:2.
Oletum Trogopterori extract of the present invention and Pollen Tyjphae extract are respectively the water extract of Oletum Trogopterori, Pollen Typhae or the extractive with organic solvent dissolving each other with water; Or, be respectively the extract that Oletum Trogopterori, Pollen Typhae obtain after water extract-alcohol precipitation is processed.
Further, the described organic solvent dissolving each other with water is selected from one or more in methanol, ethanol, acetone; Further be preferably the ethanol of 50-90%; Further be preferably the ethanol of 60-80%; Further be preferably 70% ethanol;
Extracting method used comprises any one mode decocting in extraction, reflux, extract,, immersion extraction, supersound extraction or percolation extraction, or the combination of Different Extraction Method.
Further, described alcohol precipitation concentration is 50-90%; Further preferably alcohol precipitation concentration is 60-80%; Further described alcohol precipitation concentration is 70%.
Decoction and alcohol sedimentation technique of the present invention means in Chinese medicine water extracting liquid, adds ethanol to make to reach the alcohol amount (being alcohol precipitation concentration) that contains of regulation, and some composition dissolubility in alcoholic solution reduces separates out precipitation, filter, get alcoholic solution, reclaim ethanol and obtain extract, make water extraction liquid be able to refining method.
Experimental result shows, can suppress the growth of Hela cell when Oletum Trogopterori is alone, and suppression ratio is up to 39.63%.When Pollen Typhae is alone, to the propagation of Hela cell without obvious inhibitory action.Oletum Trogopterori and Pollen Typhae compatibility effect are when human cervical carcinoma Hela cell, show as significant inhibited proliferation, when especially Oletum Trogopterori and Pollen Typhae are in 1:1 ratio compatibility, inhibitory action is the most obvious, when the concentration of Oletum Trogopterori and Pollen Typhae is respectively 62.5～500 μ g/mL, suppression ratio >30%, can reach 69.41%.
Effect experimental example
The right research of experimental example 1 Oletum Trogopterori medicine
1 material
Human cervical carcinoma Hela cell is so kind as to give by institute of oncology, Tianjin central laboratory; RPMI-1640 culture medium, U.S. GIBCO company product;
The single medicine of Oletum Trogopterori, Pollen Typhae and Oletum Trogopterori, Pollen Typhae are all adopted preparation with the following method by the compositions of different proportion compatibility (be " medicine to "): get crude drug, soak 30min, add 8 times of water gagings decoctions three times, each 1.5 hours, filter merging filtrate, centrifugal, dry, pulverize into powder.By experiment, xeraphium need to be made into reagent the filtration sterilization of respective concentration with DMEM high glucose medium.
2 experimental techniques
The take the logarithm Hela cell of trophophase, trypsinization piping and druming are cell suspension.Centrifugal collecting cell, the cell concentration of adjusting suspension after counting is 5000/mL, is inoculated in 24 porocyte culture plates, every hole 1mL.24 orifice plates are moved into 37 ℃, 5%CO
2incubator in cultivate after 24h, change liquid dosing, establish alone group of Oletum Trogopterori, Pollen Typhae, Oletum Trogopterori and Pollen Typhae are in 1:1,2:1,1:2 ratio compatibility combination group, and negative control group.With dosage group In vitro culture in following formula conversion Oletum Trogopterori and Pollen Typhae, add drug level: C (μ g/mL)=5D (mg/Kg/d); C is experiment in vitro desired concn, and D is clinical application concentration.Take this concentration as intermediate concentration, make progress respectively, downward 2 times increase progressively and successively decrease, 6 Concentraton gradient are set continuously.In incubator, cultivate 96h, light is respectively organized cellular morphology through Microscopic observation and is changed, rear collecting cell, row trypan blue counting.
In utilization, imitate principle judgement associating drug effect: middle effect concentration (Dm), i.e. half-inhibition concentration (IC
50) calculating, according to trypan blue exclusion viable count, calculate medicine group cell fu(survival rate fraction ofunaffeeted cells) and fa(suppression ratio fraetion of affected cells)
The average viable count of the average viable count/matched group of fu=test group
fa=1-fu
3 statistical method
Application SPSS16.0 statistics analysis software package, experimental data is with mean ± standard deviation
represent, under the condition of testing data normal distribution and homogeneity of variance, between many groups, data relatively adopt variance analysis, and each organizes relatively checking with LSD-t of group difference.With P < 0.05 for there being statistical significance.
4 results
Trypan blue counting method detects different experiments group: when Oletum Trogopterori is alone, can suppress the growth of Hela cell, when the concentration of Oletum Trogopterori is 125～500 μ g/mL, suppression ratio >20%, is up to 39.63%.The trend that has reduction with its suppression ratio of reduction of Oletum Trogopterori concentration.When Pollen Typhae is alone, the propagation of Hela cell, without obvious inhibitory action, is shown as to the slight facilitation to Hela Growth of Cells when the μ g/mL of Pollen Typhae concentration≤250.Oletum Trogopterori and Pollen Typhae are pressed respectively 1:1,1:2 and 2:1 ratio are share while acting on human cervical carcinoma Hela cell, show as significant inhibited proliferation, when especially Oletum Trogopterori and Pollen Typhae are in 1:1 ratio compatibility, inhibitory action is the most obvious, and suppression ratio can reach 69.41%.Refer to table 1-table 2.
Table 1 variable concentrations Oletum Trogopterori and Pollen Typhae are alone to the cytostatic effect Fa of Hela
Table 2 Oletum Trogopterori and Pollen Typhae are share the cytostatic effect Fa to Hela
The research that experimental example 2 Radix Paeoniae Alba are right
1 material
Human cervical carcinoma Hela cell is so kind as to give by institute of oncology, Tianjin central laboratory; RPMI-1640 culture medium, U.S. GIBCO company product;
The Radix Paeoniae Alba, Rhizoma Chuanxiong, each single medicine of Radix Rehmanniae Preparata and the Radix Paeoniae Alba are all adopted preparation with the following method with Rhizoma Chuanxiong, Radix Rehmanniae Preparata by the compositions of different proportion compatibility respectively: get crude drug, soak 30min, add 8 times of water gagings to decoct three times, each 1.5 hours, filter merging filtrate, centrifugal, dry, pulverize into powder.By experiment, xeraphium need to be made into reagent the filtration sterilization of respective concentration with DMEM high glucose medium.
2 experimental techniques
The take the logarithm Hela cell of trophophase, trypsinization piping and druming are cell suspension.Centrifugal collecting cell, the cell concentration of adjusting suspension after counting is 1 * 104/mL, be inoculated in 24 porocyte culture plates, every hole 1mL chooses Hela cell proliferation is had to the obvious inhibiting Radix Paeoniae Alba according to 2.1 experimental results, itself and other drug are used and act on Hela cell by different proportion variable concentrations compatibility, be equipped with the Radix Paeoniae Alba and Radix Rehmanniae Preparata in 1:1,2:1,1:2 ratio compatibility combination group, the Radix Paeoniae Alba and Rhizoma Chuanxiong in 1:1,2:1,1:2 ratio compatibility combination group, and negative control group.With dosage group In vitro culture in the following formula conversion Radix Paeoniae Alba and different pharmaceutical compatibility, add drug level: C (μ g/mL)=5D (mg/Kg/d); C is experiment in vitro desired concn, and D is clinical application concentration.Take this concentration as intermediate concentration, make progress respectively, downward 2 times increase progressively and successively decrease, 6 Concentraton gradient are set continuously.In incubator, cultivate 96h, light is respectively organized cellular morphology through Microscopic observation and is changed, rear collecting cell, row trypan blue counting.
In utilization, imitate principle judgement associating drug effect: middle effect concentration (Dm), i.e. half-inhibition concentration (IC
50) calculating, according to trypan blue exclusion viable count, calculate medicine group cell fu(survival rate fraction of unaffeeted cells) and fa(suppression ratio fraetion of affected cells)
The average viable count of the average viable count/matched group of fu=test group
fa=1-fu
Being taken the logarithm in middle efficacious prescriptions journey fa/fu=(D/Dm) m both sides, (D is drug level; Dm is middle effect concentration) equation:
log(fa/fu)=mLogD-mLogDm
Thereby Dx=Dm * [fa/ (l-fa)]
l/m
If Y=log (fa/fu), X=logD, obtains linear equation:
The slope of Y=mX+a(m equation, a is equation intercept)
During Fa=0.5, X=logDm=-a/m, the Dm value while calculating the alone and use in conjunction of medicine
Applying this formula Dx=Dm * [fa/ (1-fa)] 1//m calculates and reaches various effects (fa=0.1,0.2,0.3 ... 0.9) the alone and needed drug level of use in conjunction (D) of two kinds of medicines time
CI (The Combination index) value when calculating two medicines and share according to the amount-effect formula in week-Te Shi association index method
Clx=D1/Dxl+D2/Dx2
Attention: Dl in formula, D2 when share, the shared concentration of two medicines when effect reaches X, Dxl, Dx2 was two prescription used times, concentration separately when effect reaches X.In factor alpha basis, effect mapping is determined, when two medicines mappings are parallel to each other, α value is 0, otherwise is 1.
Result is judged: CI<l represents that when two medicines share, producing collaborative (synergism) acts on; CI=1 represents that when two medicines share, producing addition (additivity) acts on; CI>l represents that two medicines produce antagonism (antagonism) effect while share.
Middle effect concentration and associating drug effect are by CalcuSyn computed in software matching chart.
3 statistical method
Application SPSS16.0 statistics analysis software package, experimental data is with mean ± standard deviation
represent, under the condition of testing data normal distribution and homogeneity of variance, between many groups, data relatively adopt variance analysis, and each organizes relatively checking with LSD-t of group difference.With P < 0.05 for there being statistical significance.
4 experimental results
4.1 Radix Paeoniae Albas and the impact of Radix Rehmanniae Preparata compatibility on Hela cell proliferation
Trypan blue counting method detects different experiments group: the Radix Paeoniae Alba and Radix Rehmanniae Preparata are pressed 1:1, and 1:2 and 2:1 compatibility share the viable count after human cervical carcinoma Hela cell's strain 96h, calculate the cell inhibitory rate under each concentration of each experimental group.Application Excel and the CalcuSyn computed in software Radix Paeoniae Alba and Radix Rehmanniae Preparata share when different effect association index value (CI) statistical analysis of Hela cell are shown, the Radix Paeoniae Alba and Radix Rehmanniae Preparata are pressed different proportion compatibility, when 0.1<Fa<0.9, the propagation of Hela cell have been suppressed to antagonism (CI>1), wherein only when the Radix Paeoniae Alba 125 μ g/mL+ Radix Rehmanniae Preparata 62.5 μ g/mL, play slight synergism CI=0.983 (0.1<Fa<0.9) (referring to table 3-table 5).
The table 3 variable concentrations Radix Paeoniae Alba and Radix Rehmanniae Preparata are alone to the cytostatic effect Fa of Hela
Table 4 Radix Paeoniae Alba and Radix Rehmanniae Preparata share the cytostatic effect Fa to Hela
Table 5 Radix Paeoniae Alba and the CI index of Radix Rehmanniae Preparata different proportion compatibility to Hela Growth of Cells
4.2 Radix Paeoniae Albas and the impact of Rhizoma Chuanxiong compatibility on Hela cell proliferation
Trypan blue exclusion method detects different experiments group and acts on the viable count after Hela cell 96h, calculates cell inhibitory rate.Each combination group of application CalcuSyn computed in software CI shows: the Radix Paeoniae Alba and Rhizoma Chuanxiong are pressed 1:1, during 2:1 ratio compatibility, Hela cell are played to antagonism inhibitory action (CI>1).When the Radix Paeoniae Alba and Rhizoma Chuanxiong 1:2 compatibility, slight synergism when the Radix Paeoniae Alba 125 μ g/mL+ Rhizoma Chuanxiong 250 μ g/mL only, CI=0.96 (0.1<Fa<0.9), more than be antagonistic effect (CI>1) (refer to table 6-8).
The table 6 variable concentrations Radix Paeoniae Alba and Rhizoma Chuanxiong are alone to the cytostatic effect Fa of Hela
Table 7 Radix Paeoniae Alba and Rhizoma Chuanxiong share the cytostatic effect Fa to Hela
Table 8 Radix Paeoniae Alba and the CI index of Rhizoma Chuanxiong different proportion compatibility to Hela Growth of Cells
The research that experimental example 3 Elsholtzias are right
1 material
Human cervical carcinoma Hela cell is so kind as to give by institute of oncology, Tianjin central laboratory; RPMI-1640 culture medium, U.S. GIBCO company product;
Radix Et Rhizoma Rhei, Semen Persicae, each single medicine of Hirudo and Radix Et Rhizoma Rhei are all adopted preparation with the following method with Semen Persicae, Hirudo by the compositions of different proportion compatibility respectively: get crude drug, soak 30min, add 8 times of water gagings to decoct three times, each 1.5 hours, filter merging filtrate, centrifugal, dry, pulverize into powder.By experiment, xeraphium need to be made into reagent the filtration sterilization of respective concentration with DMEM high glucose medium.
2 experimental techniques
Hela cell is cultivated in 5%CO2 incubator, and culture medium is the RPMI-l640 containing l0% deactivation calf serum, containing 1% dual anti-(penicillin and streptomycin).Within every 2～3 days, go down to posterity once, the trophophase cell of taking the logarithm is for experiment.5000/m1 Hela cell of the every hole inoculation of 24 orifice plate, inoculating dosing in latter 24 hours cultivates, choose Hela cell proliferation is had to obvious inhibiting Radix Et Rhizoma Rhei, itself and other drug are used and act on Hela cell by different proportion variable concentrations compatibility, be equipped with alone group of Radix Et Rhizoma Rhei, Semen Persicae, Hirudo, Radix Et Rhizoma Rhei and Semen Persicae in 1:1,2:1,1:2 ratio compatibility combination group, Radix Et Rhizoma Rhei and Hirudo in 1:1,2:1,1:2 ratio compatibility combination group, and negative control group.With dosage group In vitro culture in following formula conversion Radix Et Rhizoma Rhei and different pharmaceutical compatibility, add drug level: C (μ g/mL)=5D (mg/Kg/d); C is experiment in vitro desired concn, and D is clinical application concentration.Take this concentration as intermediate concentration, make progress respectively, downward 2 times increase progressively and successively decrease, 6 Concentraton gradient are set continuously.In incubator, cultivate 96h, light is respectively organized cellular morphology through Microscopic observation and is changed, rear collecting cell, row trypan blue counting.
In utilization, imitate principle judgement associating drug effect: middle effect concentration (Dm), be the calculating of half-inhibition concentration (IC50), according to trypan blue exclusion viable count, calculate medicine group cell fu(survival rate fraction of unaffeeted cells) and fa(suppression ratio fraetion of affected cells)
The average viable count of the average viable count/matched group of fu=test group
fa=1-fu
Being taken the logarithm in middle efficacious prescriptions journey fa/fu=(D/Dm) m both sides, (D is drug level; Dm is middle effect concentration) equation:
log(fa/fu)=mLogD-mLogDm
Thereby Dx=Dm * [fa/ (l-fa)] l/m
If Y=log (fa/fu), X=logD, obtains linear equation:
The slope of Y=mX+a(m equation, a is equation intercept)
During Fa=0.5, X=logDm=-a/m, the Dm value while calculating the alone and use in conjunction of medicine
Applying this formula Dx=Dm * [fa/ (1-fa)] 1//m calculates and reaches various effects (fa=0.1,0.2,0.3 ... 0.9) the alone and needed drug level of use in conjunction (D) of two kinds of medicines time
CI (The Combination index) value when calculating two medicines and share according to the amount-effect formula in week-Te Shi association index method
Clx=D1/Dxl+D2/Dx2
Attention: Dl in formula, D2 when share, the shared concentration of two medicines when effect reaches X, Dxl, Dx2 was two prescription used times, concentration separately when effect reaches X.In factor alpha basis, effect mapping is determined, when two medicines mappings are parallel to each other, α value is 0, otherwise is 1.
Result is judged: CI<l represents that when two medicines share, producing collaborative (synergism) acts on; CI=1 represents that when two medicines share, producing addition (additivity) acts on; CI>l represents that two medicines produce antagonism (antagonism) effect while share.
Middle effect concentration and associating drug effect are by CalcuSyn computed in software matching chart.
3 statistical method
Application SPSS16.0 statistics analysis software package, experimental data is with mean ± standard deviation
represent, under the condition of testing data normal distribution and homogeneity of variance, between many groups, data relatively adopt variance analysis, and each organizes relatively checking with LSD-t of group difference.With P < 0.05 for there being statistical significance.
4 experimental results
4.1 Radix Et Rhizoma Rhei and the impact of Semen Persicae compatibility on Hela cell proliferation
Radix Et Rhizoma Rhei and Semen Persicae are alone and share (Radix Et Rhizoma Rhei: Semen Persicae=1:1,1:2 and 2:1) viable count after human cervical carcinoma Hela cell's strain 96h, calculate the cell inhibitory rate under each concentration of each experimental group.Application Excel and CalcuSyn computed in software Radix Et Rhizoma Rhei and Semen Persicae share when different effect association index value (CI) statistical analysis of Hela cell are shown, Radix Et Rhizoma Rhei and Semen Persicae have suppressed antagonism (CI>1) (refer to show 10-table 12) to the propagation of Hela cell by different proportion compatibility when the 0.1<Fa<0.9.
Table 10 variable concentrations Radix Et Rhizoma Rhei and Semen Persicae are alone to the cytostatic effect Fa of Hela
Table 11 Radix Et Rhizoma Rhei and Semen Persicae share the cytostatic effect Fa to Hela
Table 12 Radix Et Rhizoma Rhei and the CI index of Semen Persicae different proportion compatibility to Hela Growth of Cells
4.2 Radix Et Rhizoma Rhei and the impact of Hirudo compatibility on Hela cell proliferation
Each combination group of application CalcuSyn computed in software CI shows: Radix Et Rhizoma Rhei and Hirudo are pressed 1:1, and 2:1 plays antagonism inhibitory action (CI>1) (referring to table 13-table 15) to Hela cell during 1:2 ratio compatibility.
Table 13 variable concentrations Radix Et Rhizoma Rhei and Hirudo are alone to the cytostatic effect Fa of Hela
Table 14 Radix Et Rhizoma Rhei and Hirudo are share the cytostatic effect Fa to Hela
Table 15 Radix Et Rhizoma Rhei and the CI index of Hirudo different proportion compatibility to Hela Growth of Cells
The specific embodiment
Embodiment 1 oral liquid
Prescription crude drug forms: Oletum Trogopterori 30g, Pollen Typhae 30g;
Get the crude drug of recipe quantity, decoct with water 2 times, 2 hours for the first time, 1.5 hours for the second time, collecting decoction, filters, and filtrate is concentrated, concentrated solution adds ethanol to containing alcohol amount to 70%, standing, filter, filtrate recycling ethanol is extremely without alcohol taste, and to be concentrated into relative density be 1.15(60 ℃), add conventional adjuvant to make oral liquid.
Embodiment 2 tablets
Prescription crude drug forms: Oletum Trogopterori 30g, Pollen Typhae 15g;
Get the crude drug of recipe quantity, add 5 times of amount 70% alcohol reflux 2 times, each 1.5 hours; Merge extractive liquid,, filters, and reclaims ethanol concentrated; Concentrated solution drying under reduced pressure, is ground into fine powder, adds conventional adjuvant, mixes; Dry-pressing is granulated, and tabletting, obtains.
Embodiment 3 capsules
Prescription crude drug forms: Oletum Trogopterori 30g, Pollen Typhae 60g;
Get the crude drug of recipe quantity, add 10 times of amount methanol supersound extraction 3 times, 40min for the first time, the 2nd, 3 each 20min, merge extractive liquid,, filters concentrating under reduced pressure; Concentrated solution drying under reduced pressure, is ground into fine powder, adds conventional adjuvant, mixes; Dry-pressing is granulated, and incapsulates, and obtains.
Embodiment 4 granules
Prescription crude drug forms: Oletum Trogopterori 30g, Pollen Typhae 120g;
Get the crude drug of recipe quantity, soak 30min, add 8 times of water gagings to decoct three times, each 1.5 hours, filter, merging filtrate, concentrated; Concentrated solution, by macroporous resin, is first used 5 times of water gaging eluting, and eluent is abandoned or adopted, then with adding 70% ethanol elution, collects eluent, and decompression recycling ethanol is extremely without alcohol taste; 80 ℃ of following drying under reduced pressure, are ground into fine powder, add adjuvant, mix; Dry-pressing is granulated, and obtains.
Embodiment 5 oral liquids
Prescription crude drug forms: Oletum Trogopterori 30g, Pollen Typhae 90g;
Get recipe quantity crude drug, decoct with water 2 times, each 1.5 hours, collecting decoction, filtered, filtrate is concentrated, and concentrated solution adds ethanol to containing alcohol amount to 80%, standing, filters, filtrate recycling ethanol is to without alcohol taste, and to be concentrated into relative density be 1.15(60 ℃), add adjuvant, make oral liquid.
Embodiment 6 drop pill
Prescription crude drug forms: Oletum Trogopterori 30g, Pollen Typhae 9g;
Get the crude drug of recipe quantity, add 8 times of water gaging dipping 6h, percolation 24h, flow velocity 2L/h.Collect percolate, centrifugal, upper macroporous adsorptive resins, first washes with water, then uses 60% ethanol elution, collects ethanol elution, and filtrate is concentrated, then adds acceptable adjuvant in pharmacy to make drop pill according to the routine techniques of this area preparation.
Embodiment 7 tablets
Prescription crude drug forms: Oletum Trogopterori 30g, Pollen Typhae 6g;
Get the crude drug of recipe quantity, add 5 times of amount 60% alcohol reflux 2 times, each 1.5 hours; Merge extractive liquid,, filters, and reclaims ethanol concentrated; Concentrated solution drying under reduced pressure, is ground into fine powder, adds conventional adjuvant, mixes; Dry-pressing is granulated, and tabletting, obtains.
Embodiment 8
Prescription crude drug forms: Oletum Trogopterori extract 30g, Pollen Tyjphae extract 30g;
Described Oletum Trogopterori extract, Pollen Tyjphae extract are respectively the extract that Oletum Trogopterori, Pollen Typhae prepare through 60% alcohol reflux.By said extracted thing drying under reduced pressure, pulverize as fine powder, mix, add the agent of conventional adjuvant granulation.
Embodiment 9
Prescription raw material forms: Oletum Trogopterori extract 30g, Pollen Tyjphae extract 15g;
Described Oletum Trogopterori extract, Pollen Tyjphae extract are respectively Oletum Trogopterori, Pollen Typhae and through decocting, boil the extract preparing.By said extracted thing drying under reduced pressure, pulverize as fine powder, mix, add conventional adjuvant to make tablet.
Embodiment 10
Prescription raw material forms: Oletum Trogopterori extract 30g, Pollen Tyjphae extract 60g;
Described Oletum Trogopterori extract, Pollen Tyjphae extract are respectively the extract that Oletum Trogopterori, Pollen Typhae prepare through acetone reflux, extract.By said extracted thing drying under reduced pressure, pulverize as fine powder, mix, add conventional adjuvant to make capsule.
Embodiment 11
Prescription raw material forms: Oletum Trogopterori extract 30g, Pollen Tyjphae extract 90g;
Described Oletum Trogopterori extract, Pollen Tyjphae extract are respectively the extract that Oletum Trogopterori, Pollen Typhae prepare through aqueous extraction-alcohol precipitation technology, and its alcohol precipitation concentration is that 70% alcoholic solution percolation extracts the extract obtaining; By said extracted thing drying under reduced pressure, be ground into fine powder, add conventional adjuvant, mix; Dry-pressing is granulated, and tabletting, obtains.
Claims (10)
1. a Chinese medicine composition with antitumaous effect, is characterized in that, the crude drug of this Chinese medicine composition is mainly by Oletum Trogopterori, Pollen Typhae.
2. Chinese medicine composition as claimed in claim 1, is characterized in that, this Chinese medicine composition is comprised of the crude drug of following weight ratio:
Oletum Trogopterori: Pollen Typhae=1:(0.2-4);
Or, Oletum Trogopterori: Pollen Typhae=1:(0.3-3.5);
Or, Oletum Trogopterori: Pollen Typhae=1:(0.4-3);
Or, Oletum Trogopterori: Pollen Typhae=1:(0.5-2.5);
Or, Oletum Trogopterori: Pollen Typhae=1:(0.5-2.2).
3. Chinese medicine composition as claimed in claim 2, is characterized in that, this Chinese medicine composition is comprised of the crude drug of following weight ratio:
Oletum Trogopterori: Pollen Typhae=1:0.5;
Or, Oletum Trogopterori: Pollen Typhae=1:1;
Or, Oletum Trogopterori: Pollen Typhae=1:2.
4. as described in as arbitrary in claim 1-3, the preparation method of Chinese medicine composition, is characterized in that, the method comprises:
Get in proportion crude drug, with water or the organic solvent extraction that dissolves each other with water;
Or, get in proportion crude drug, adopt decoction and alcohol sedimentation technique preparation; Wherein said alcohol precipitation concentration is 50-90%.
5. the preparation method of Chinese medicine composition as claimed in claim 4, is characterized in that the described organic solvent dissolving each other with water is selected from one or more in methanol, ethanol, acetone; Described extracting method comprises any one mode decocting in extraction, reflux, extract,, immersion extraction, supersound extraction or percolation extraction, or the combination of Different Extraction Method.
6. a Chinese medicine composition with antitumaous effect, is characterized in that, this Chinese medicine composition is comprised of the raw material of following weight ratio:
Oletum Trogopterori extract: Pollen Tyjphae extract=1:(0.2-4).
7. Chinese medicine composition as claimed in claim 6, is characterized in that, this Chinese medicine composition is comprised of the raw material of following weight ratio:
Oletum Trogopterori extract: Pollen Tyjphae extract=1:(0.3-3.5);
Or, Oletum Trogopterori extract: Pollen Tyjphae extract=1:(0.4-3);
Or, Oletum Trogopterori extract: Pollen Tyjphae extract=1:(0.5-2.5);
Or, Oletum Trogopterori extract: Pollen Tyjphae extract=1:(0.5-2.2).
8. Chinese medicine composition as claimed in claim 7, is characterized in that, this Chinese medicine composition is comprised of the raw material of following weight ratio:
Oletum Trogopterori extract: Pollen Tyjphae extract=1:0.5;
Or, Oletum Trogopterori extract: Pollen Tyjphae extract=1:1;
Or, Oletum Trogopterori extract: Pollen Tyjphae extract=1:2.
9. the application of the Chinese medicine composition as described in as arbitrary in claim 1-3,6-8 in preparation treatment cancer drug.
10. the application of the Chinese medicine composition as described in as arbitrary in claim 1-3,6-8 in preparation treatment cervical cancer medicine.
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Non-Patent Citations (3)
| Title |
|---|
| 王付等主编: "《方剂学（第二版）》", 31 August 2012, 中国中医药出版社 * |
| 程昭寰主编: "《方剂气味配伍理论及应用》", 31 January 2006, 中国中医药出版社 * |
| 郑大坤主编: "《中医药临床速查手册》", 31 January 2007, 江苏科学技术出版社 * |
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