CN101099769B - Application of Fructus Forsythiae in preparing anti-tumor chemotherapeutic sensitivity intensifying attenuating medicine - Google Patents
Application of Fructus Forsythiae in preparing anti-tumor chemotherapeutic sensitivity intensifying attenuating medicine Download PDFInfo
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Abstract
The present invention relates to an application of Chinese medicinal material forsythia fruit in preparation of anti-tumor chemotherapeutic sensitivity-enhancing and attenuating medicine. It is characterized by that the invented medicine can be made up by using forsythia fruit alcohol extract and medicinal carrier and/or excipient through a certain preparation process.
Description
Technical field
The present invention relates to the purposes of Fructus Forsythiae, relate in particular to the purposes in pharmaceutical field.
Background technology
Add up the eighties in 20th century according to World Health Organization (WHO), annual about 7,000,000 people of whole world pathogenesis of cancer number, about 5,000,000 people of the annual number of cancer mortality.To the nineties, annual about 1,000 ten thousand people of whole world pathogenesis of cancer number, annual dead about 7,000,000 people.Cancer is on the rise in the harm in the whole world.In coming decade, it is first of the disease of serious harm human health that cancer can replace that the cardiovascular diseases becomes.At China's the mid-1970s, treatment and prevention of tumour research office in the Ministry of Public Health whole nation organizes national cancer mortality retrospective survey, at that time, and annual about 900,000 people of pathogenesis of cancer, annual about 700,000 people of cancer mortality.To the initial stage nineties, according to the national treatment and prevention of tumour research sampling survey result of office, annual about 1,600,000 people in the pathogenesis of cancer whole nation, dead annual about 1,300,000 people.At present, cancer has become first cause of death of China urban and rural residents.
Chemotherapy is an important means for the treatment of cancer at present, and also there is toxicity in chemotherapeutics to organism normal cell in kill cancer cell.In recent years, along with continuing to bring out of PTS, the rational Application of Combination chemotherapy, the curative effect of chemotherapy is enhanced, but these medicines have infringement or toxic action to normal histoorgan simultaneously, thereby reduce body's immunity, a series of toxicities such as bone marrow depression occur, influence the curative effect of chemotherapy, even force chemotherapy to stop.Clinical research shows, when chemotherapy,, be aided with treatment by Chinese herbs, can strengthen the effect of chemotherapy effectively according to the toxicity symptom of patient's body constitution, cancer staging and appearance, the toxicity of chemicotherapy be can avoid or alleviate again to a certain extent, survival time of patients and life quality prolonged.And Chinese medicine has special advantages as the advantageous resource of China aspect the chemotherapy of tumors efficacy enhancing and toxicity reducing, and it can promote hematopoietic function recovery; Human body immunity improving function again; Can also prevent and treat gastrointestinal reaction; Some medicine also has obvious antineoplastic, and therefore the medicine that screening has the chemotherapy attenuation synergistic from the traditional natural drug of China has crucial meaning.
Fructus Forsythiae is the dry fruit of Oleaceae forsythia Fructus Forsythiae, and bitter in the mouth is nontoxic, and cold nature has effects such as heat clearing away emesis, liver heat removing function of gallbladder promoting, dehumidifying jaundice eliminating, logical accent three warmers, fluent blood vessels, protecting liver and heart, and clinical practice is extensive., but share about Fructus Forsythiae and chemotherapeutic, increase the chemotherapeutic curative effect, reduce the toxic research of chemotherapeutic and there is no relevant report, the invention provides the Forsythinol extract as a kind of new chemotherapeutic sensitizer, be used for the antineoplastic auxiliary treatment.
Summary of the invention
The present invention provides the purposes of Fructus Forsythiae in preparation antineoplastic adjuvant therapy medicaments at the problems referred to above, with the medicine of Fructus Forsythiae preparation the antitumor action of chemotherapeutic is had obvious role in synergism, and can reduce side effect such as hemocyte decline that chemotherapeutic causes, liver, injury of kidney.
Technical scheme provided by the invention is: the application of Fructus Forsythiae in preparation anti-tumor chemotherapeutic enhanced sensitivity toxicity-reducing medicament.
Above-mentioned anti-tumor chemotherapeutic enhanced sensitivity toxicity-reducing medicament is the compositions of Forsythinol extract and the pharmaceutical carrier and/or the excipient of effective dose.
Above-mentioned anti-tumor chemotherapeutic enhanced sensitivity toxicity-reducing medicament is an oral formulations.
Above-mentioned anti-tumor chemotherapeutic enhanced sensitivity toxicity-reducing medicament is an injection.
The present invention can make Pharmaceutical composition with Forsythinol extract and pharmaceutical carrier and/or excipient.Prepared Pharmaceutical composition is used for the auxiliary treatment of anti-tumor chemotherapeutic, has good effect, advantages such as few side effects.Advise clinical consumption: one day twice oral, each 50-200mg Forsythinol extract.
Description of drawings
Fig. 1 is the HE stained figure of Forsythinol extract to the influence of rat liver tectology;
Fig. 2 is the HE stained figure of Forsythinol extract to the influence of rat kidney tectology.
The specific embodiment
The present invention is described further by following examples
Embodiment 1: compositions is restrained into 1000 by Forsythinol extract 1～100 gram, cellulose 400～499.Its preparation method is: Forsythinol extract and cellulose granulated, and tabletting or fill capsule then, every (sheet) contains Forsythinol extract 1-100mg.
Embodiment 2: the Forsythinol extractive composition adds injection water to 1 liter by 1～100 gram Forsythinol extract, is distributed into 1000 and makes.Its method for making is: the Forsythinol extract is dissolved in the water for injection, adds to 1 liter, sterilization, packing.Every contains 1-100mg Forsythinol extract.
The Forsythinol extract of the foregoing description can be obtained by conventional extracting method; As the alcohol reflux that Fructus Forsythiae added 3 times of volumes 70% 2 hours, repeat 3 times, merge extractive liquid,, refluxing to concentrate obtains the Forsythinol extract.
The external synergistic antitumor effect research of Fructus Forsythiae active component
Experimental technique
The experiment grouping
CY+DDP (cisplatin+cyclophosphamide) processed group
(1) normal control group (control); (2) model group (model, CY+DDP, 60 μ gml
-1+ 10 μ gml
-1); Each dosage group (CY+DDP+LQ) of Forsythinol extract (LQ) is respectively: (3) LQ1 (CY+DDP+LQ 100 μ gml
-1), (4) LQ2 (CY+DDP+LQ 50 μ gml
-1), (5) LQ3 (CY+DDP+LQ 10 μ gml
-1), (6) LQ4 (CY+DDP+LQ 2 μ gml
-1), (7) LQ5 (CY+DDP+LQ0.4 μ gml
-1); All drug level are final concentration.
Without the CY+DDP processed group
(1) normal control group (control); Each dosage group (LQ) of Forsythinol extract is respectively: (2) LQ1 (100 μ gml
-1), (3) LQ2 (50 μ gml
-1), (4) LQ3 (10 μ gml
-1), (5) LQ4 (2 μ gml
-1), (6) LQ5 (0.4 μ gml
-1); All drug level are final concentration.
Mtt assay
Collect the cell of exponential phase, plant in 96 well culture plates. every hole 1 * 10
4Individual cell, put 5%CO2 incubator 24h after, add medicine. behind 48h, remove culture fluid, every hole add 100 μ g MTT[3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazoliumbromide], 37 ℃ of incubation 4h, remove supernatant, add 150ul DMSO dissolving, vibration 10min mixing, measure the light absorption value (A) of wavelength 490nm, calculate suppression ratio.
Medication combined evaluation of effect
The computing formula of two medicine interaction indexes (coefficient of drug interaction.CDI) is: and CDI=AB/ (A * B).In vitro tests is to calculate according to viable count (light absorption value).The interior therapeutic test is heavily to calculate according to tumor.AB is the ratio of two medicine coupling groups and matched group light absorption value; A or B are the ratio of each medicine use group separately and matched group light absorption value.Two medicines have synergism when CDI＜1; When CDI＜0.7, the synergism highly significant; As CDI=1, then two medicine interaction properties are addition; As CDI＞1, then two medicine interaction properties are antagonism.
2.4
Data represent that with x ± S mean relatively adopts the t check, relatively adopt variance analysis between many groups, and data analysis adopts SPSS 10.0 statistical softwares.
Experimental result
1 Fructus Forsythiae active component and CY+DDP suppress the synergism of growth of tumour cell
Experimental result shows, use separately CY+DDP that the suppression ratio of gastric carcinoma cells SGC2 and Mus transplanted hepatoma cell H22 is respectively 68.91% and 77.33%, and during with Forsythinol extract synergism, suppression ratio obviously improve, CDI has synergism all less than 1; Phillyrin and CY+DDP share the inhibitory action of tumor cell obvious, and CDI is all less than 1, and suppression ratio increases with the increase with dosage.
The collaborative influence to the SGC2 cell proliferation of table 1 Fructus Forsythiae active component and CY+DDP (x ± S, n=6)
**CDI＜0.7，0.7＜*CDI＜1
The collaborative influence to the H22 cell proliferation of table 2. Fructus Forsythiae active component and CY+DDP (x ± S, n=6)
**CDI＜0.7，0.7＜*CDI＜1
3. the Fructus Forsythiae active component is to the influence of SGC2, the growth of H22 cell
Compare with the blank group, the Forsythinol extract does not all make significant difference to SGC2, the growth of H22 cell, the results are shown in Table 3
Table 3 Fructus Forsythiae active component to the influence of SGC2, H22 cell proliferation (x ± S, n=6)
Experimental result
Experimental result shows, use separately CY+DDP that the suppression ratio of gastric carcinoma cells SGC2 and Mus transplanted hepatoma cell H22 is respectively 68.91% and 77.33%, and during with Forsythinol extract synergism, suppression ratio obviously improve, CDI has synergism all less than 1; Phillyrin and CY+DDP share the inhibitory action of tumor cell obvious, and CDI is all less than 1, and suppression ratio increases with the increase of dosage.
Fructus Forsythiae is to the effect of chemotherapeutic efficacy enhancing and toxicity reducing
Experimental technique
Animal grouping and processing
Healthy male SD rat y60, normal feedstuff is fed a week, as the laundering period, is divided into 6 groups at random, and 10 every group, normal diet feeding, 20g//day, totally 7 days.Modeling method: beginning modeling in first day, administration simultaneously, every rats by intraperitoneal injection cyclophosphamide (60mg/kg) and cisplatin (10mg/kg), continuous three days.Normal group mice then injecting normal saline is contrast.Grouping: normal group (ig. normal saline); Model group (CY+DDP, ig. normal saline); The Fructus Forsythiae high dose group (CY+DDP+LQ1, ig.400mg/kg, qd); Dosage group in the Fructus Forsythiae (CY+DDP+LQ2, ig.80mg/kg, qd); The Fructus Forsythiae low dose group (CY+DDP+LQ3, ig.16mg/kg, qd); Positive controls (CY+DDP+LTN, ig.5mg/kg, qd)
Put to death animal after 7 days, the eye socket blood sampling divides two pipes, and wherein a pipe heparin sodium anticoagulant is respectively applied for biochemical function and detects and whole blood cell analysis.Win immune organ spleen, thymus, fix in 4% paraformaldehyde, paraffin embedding, section, about 4 μ m are thick, HE dyeing, light microscopic procuratorial work; Win liver, nephridial tissue, as for fixing in 5% glutaraldehyde, do transmission electron microscope observing rapidly.
Influence to the rat complete blood cell
After the administration 7 days, the animal fasting be can't help using etherization behind the water 12h, and the eye socket blood sampling places 1.5ml EP pipe (adding the heparin sodium anticoagulant), carries out blood cell analysis immediately.
Influence to rats'liver function and hepatic pathology variation
Influence to the rats'liver function
The eye socket blood sampling places the 8mlEP pipe, and the centrifugal 10min of 3000rpm gets upper serum in the EP pipe, carries out the detection of serum biochemistry index.
Influence to the rat liver pathological change
1. perusal
Win whole liver behind the sacrifice of animal, perusal liver form, size, color and luster and quality.
2. mirror is observed down
Get same place lobe of the liver sheet and fix, make the hepatic tissue pathology section routinely, carry out HE dyeing respectively and under light microscopic, check with 10% paraformaldehyde.
Influence to kidney of rats function and kidney pathological change
Influence to the kidney of rats function
The eye socket blood sampling places the 5mlEP pipe, and the centrifugal 10min of 3000rpm gets upper serum in the EP pipe, carries out the detection of serum biochemistry index.
Influence to the rat kidney pathological change
1. perusal
Win the both sides kidney behind the sacrifice of animal, its form of perusal, size, color and luster and quality.
2. mirror is observed down
Get the same side kidney and fix, make the nephridial tissue pathological section routinely, carry out HE dyeing respectively and under light microscopic, check with 10% paraformaldehyde.
The foundation of ascites tumor model and animal are handled
Get this chamber 5～7d that goes down to posterity voluntarily, well-grown milky ascites is diluted to tumor cell suspension with an amount of physiological saline solution, and cell number is 5 * 10
6Individual, only inoculate with 0.2ml/.Mouse inoculation 24h is divided into 5 groups later at random, 8 every group.Be divided into: (1) blank group (control); The high group of administering drug combinations group (2) Fructus Forsythiae (CY+DDP+LQ1, ig.400mg/kg, qd); (3) organize in the Fructus Forsythiae (CY+DDP+LQ2, ig.80mg/kg, qd); (4) Fructus Forsythiae is low organizes (CY+DDP+LQ3, ig.16mg/kg, qd); Positive group (DDP+CY, ip.1mg/kg+10mg/kg, qd).Put to death animal after 8 days, pluck the eyeball blood sampling, divide two pipes, wherein a pipe heparin sodium anticoagulant is respectively applied for biochemical function and detects and whole blood cell analysis.The extraction mouse ascites is weighed, simultaneously the ascites cells counting.
Statistical disposition
(x ± S) expression, relatively checking with t of two sample means relatively adopted variance analysis between many groups to experimental data mean scholar standard deviation, data analysis employing SPSS 10.0 statistical softwares.
Influence to the rat complete blood cell
By table 4 as seen, model group whole blood WBC (leukocyte), PLT (platelet), RTC% (reticulocyte ratio) level are compared obvious reduction with matched group, leucocyte level is compared obvious rising in Fructus Forsythiae height, middle dosage group, the positive group whole blood with model group, and each administration group RBC (erythrocyte), HGB (hemoglobin) compare no significant difference with model group.
The influence of table 4 pair rat complete blood cell (x ± S, n=10)
Compare * P＜0.05, * * P＜0.01 with model group; Compare with normal group,
▲P＜0.05,
▲ ▲P＜0.01. is to the influence of rats'liver, renal function
By table 5 as seen, model group plasma A LT (glutamate pyruvate transaminase), AST (glutamic oxaloacetic transaminase, GOT), BUN (blood urea nitrogen), CREA (inosine clearance rate) level are compared remarkable rising (P＜0.01) with matched group; Compare with model group, Fructus Forsythiae height, middle dosage group and positive group ALT, BUN level obviously reduce (P＜0.05), and wherein dosage group serum AST level is compared obvious reduction (P＜0.05) than model group in the Fructus Forsythiae; Fructus Forsythiae height, middle dosage group are compared with model group, and change of serum C REA level obviously reduces (P＜0.05).
The influence of table 5 pair rats'liver, renal function (x ± S, n=10)
Compare * P＜0.05 with model group, * * P＜0.01; Compare with normal group
▲P＜0.05,
▲ ▲P＜0.01.
Influence to ascites tumor mouse ascites growing amount and oncocyte number
By table 6 as seen, compare with matched group, each dosage group of Fructus Forsythiae all is remarkable decline (P＜0.01) with positive group ascites growing amount and oncocyte quantity, wherein in the Fructus Forsythiae, that low dosage drug combination group is counted the minimizing effect to ascites tumor mouse ascites growing amount and oncocyte is obvious.
Table 6 Forsythinol extract to the influence of ascites tumor mouse ascites growing amount and oncocyte number (x ± S, n=8)
Compare with matched group,
▲P＜0.05,
▲ ▲P＜0.01.
Influence to ascites tumor mice complete blood cell
By table 7 as seen, compare with matched group, each dosage group of Fructus Forsythiae and DDP+CY group whole blood WBC (leukocyte), LYM (lymphocyte), PLT (platelet) level all descend to some extent, WBC and DDP+CY group is compared on the risely in each dosage group complete blood cell of Fructus Forsythiae, and each administration group RBC (erythrocyte), HGB (hemoglobin) compare no significant difference with model group
Table 7 Forsythinol extract to the influence of ascites tumor mice complete blood cell (x ± S, n=8)
Compare with matched group
▲P＜0.05,
▲ ▲P＜0.01; Compare * P＜0.05 with DDP+CY (cisplatin+cyclophosphamide) group, * * P＜0.01. is to the influence of ascites tumor Mouse Liver, renal function
By table 8 as seen, plasma A LT, AST, BUN, ALP, the CREA level of comparing each dosage group of Fructus Forsythiae with the DDP+CY group have to a certain degree reduction, and compare each dosage treated animal plasma A LT of Fructus Forsythiae, AST, BUN, the horizontal no significant difference of CREA with the blank group.
Table 8 Forsythinol extract to the influence of ascites tumor Mouse Liver, renal function (x ± S, n=8)
Compare with matched group,
▲P＜0.05,
▲ ▲P＜0.01; With DDP+CY (cisplatin+cyclophosphamide) group * P＜0.05, * * P＜0.01.
Influence to ascites tumor Mouse Liver, the variation of nephridial tissue pathology
The rat liver liver histological is checked: compare with normal liver tissue, the visible hepatocyte cloudy swelling of model group, vacuolar degeneration are arranged more loosely, are dispersed in a large amount of fat in the kytoplasm and drip, the Forsythinol extract can obviously reduce the extent of damage of hepatic tissue, the basic normal (see figure 1) of visible lobules of liver structure.Among Fig. 1, A: matched group; B:CY+DDP (cisplatin+cyclophosphamide) group; C:CY+DDP+LQ1 (cisplatin+cyclophosphamide+Fructus Forsythiae low dosage) group; D:CY+DDP+LQ2 (dosage in cisplatin+cyclophosphamide+Fructus Forsythiae) group; E:CY+DDP+LQ3 (cisplatin+cyclophosphamide+Fructus Forsythiae high dose) group; F:CY+DDP+LTN (cisplatin+cyclophosphamide+tiopronin) group;
The rat kidney histological examination: the om observation model group is compared with normal group, and renal cells has the cloudy swelling pathological changes, and brush border has disappearance, the cellular swelling. (see figure 2).Fructus Forsythiae coupling group pathological changes is not obvious, glomerule and renal tubules structure complete substantially (seeing figure C, D, E).Among Fig. 2, A: matched group; B:CY+DDP (cisplatin+cyclophosphamide) group; C:CY+DDP+LQ1 (cisplatin+cyclophosphamide+Fructus Forsythiae low dosage) group; D:CY+DDP+LQ2 (dosage in cisplatin+cyclophosphamide+Fructus Forsythiae) group; E:CY+DDP+LQ3 (cisplatin+cyclophosphamide+Fructus Forsythiae high dose) group; F:CY+DDP+LTN (cisplatin+cyclophosphamide+tiopronin) group;
Discuss
Fructus Forsythiae active component synergistic antitumor function analysis
Fructus Forsythiae has the heat clearing away thermal detoxification, multiple pharmacological effect such as antibiotic, antiviral, antiinflammatory, analgesia, and its active ingredient phenethyl alcohol glycosides, terpenoid etc. have effects such as antitumor, antiviral.Experiment in vitro is the result show, the Forsythinol extract has synergism with chemotherapeutics cyclophosphamide and cisplatin combined use the time, and drug interaction index (CDI) is between 0.7-1.Experimental result shows that Forsythinol extract and chemotherapeutics share in the body, and the inhibition of Mus transplanted hepatoma H22 ascites tumor mouse ascites growing amount and oncocyte number is had synergism.Prompting Fructus Forsythiae and chemotherapeutics share has the synergistic antitumor effect.
The Forsythinol extract is to the protective effect analysis of liver, injury of kidney due to the chemotherapy
This experiment is further studied the effect of Forsythinol extract aspect the chemotherapy auxiliary treatment from aspects such as biochemistry, blood, tectology and Ultrastructural variations more comprehensively.Experimental result shows that the quantity of leucocyte that the Forsythinol extract can promote due to the chemotherapy reduces, and the hepatic and renal function that the protection chemotherapeutics causes damages.Point out it to have potential chemotherapeutics auxiliary treatment medicinal efficacy.
In sum; by to Forsythinol extract and active component thereof in vivo, external research; we find aspect the immunocompromised and hepatic and renal function damage of Forsythinol extract due to the protection chemotherapy; have some improvement; find also that simultaneously Fructus Forsythiae and chemotherapeutics share and have the synergistic antitumor effect, the prompting Fructus Forsythiae has the effect of the attenuation synergistic in the chemotherapeutic treatment process.
Claims (4)
1. the application of Forsythinol extract in preparation anti-tumor chemotherapeutic enhanced sensitivity toxicity-reducing medicament.
2. application according to claim 1, it is characterized in that: described anti-tumor chemotherapeutic enhanced sensitivity toxicity-reducing medicament is the compositions of Forsythinol extract, pharmaceutical carrier and the excipient of effective dose, the perhaps compositions of Forsythinol extract and excipient, the perhaps compositions of Forsythinol extract and pharmaceutical carrier.
3. application according to claim 2 is characterized in that: described anti-tumor chemotherapeutic enhanced sensitivity toxicity-reducing medicament is an oral formulations.
4. application according to claim 2 is characterized in that: described anti-tumor chemotherapeutic enhanced sensitivity toxicity-reducing medicament is an injection.
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| CN1947747A (en) * | 2005-10-10 | 2007-04-18 | 黄振华 | Traditional Chinese medicine composition contg. luteolin and capsule of sweeping forsythia and its prepn. method and use |
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| CN1879720A (en) * | 2006-05-06 | 2006-12-20 | 安徽科创中药天然药物研究所有限责任公司 | Blood platelet-increasing tablet, its preparation process and quality control method |
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