CN101623295A - Medicine for treating bladder neoplasms - Google Patents

Abstract

The invention relates to a medicine which is prepared by the compatibility and the compounding of active ingredients extracted from traditional Chinese medicines and is used for affusing the bladder and treating bladder neoplasms. The medicine is prepared by the following traditional Chinese medical monomers of panaxoside Rg3, meletin and curcumin in a mol ratio of 20:5:3, and the optimal medical composition comprises the three traditional Chinese medical monomers: 120-200 micromole/l of panaxoside Rg3, 18-30 micromole/l of meletin and 29-47 micromole/l of curcumin.

Description

The medicine that is used for the treatment of tumor of bladder

Technical field

The present invention relates to a kind of medicine, particularly a kind ofly be used in intravesical perfusion therapy tumor of bladder, form, be used for medicine in intravesical perfusion therapy tumor of bladder by the active ingredient compatibility compound recipe that from Chinese medicine, extracts.

Background technology

Tumor of bladder is the male genitourinary system tumor, and in western countries, its sickness rate is only second to carcinoma of prostate in male's genito-urinary system tumor, occupies second.And according to Epidemiological study, in the U.S., the sickness rate of this disease in the past 30 in the period of raise steadily.Tumor of bladder is also comparatively the most common in China.Because the age is one of main onset risk factor of this disease, can predict, along with increasing the weight of of aged tendency of population, the sickness rate of tumor of bladder will further raise.

Be the shallow tumor more than 80% in the tumor of bladder, present stage, main Therapeutic Method was an operative treatment, and postoperative is aided with the intravesical dabbling drug and attacks to the flesh layer to suppress tumor recurrence and to delay it.If postoperative does not carry out auxiliary treatment, the relapse rate in 1 year of postoperative is up to 80%.Since Morales in 1976 is applied to bacillus calmette-guerin vaccine (BCG) superficial bladder tumor postoperative perfusion prevention of recurrence first, the BCG irrigation of bladder has become a kind of important means of tumor of bladder auxiliary treatment, and BCG is the intravesical dabbling drug of the unique approval of U.S. FDA at present.But its definite antitumor mechanism it be unclear that so far, and BCG intravesical perfusion toxic and side effects is big, and the patient is difficult to adhere to finishing whole perfusion course of treatment, thereby affects the treatment.

Chinese invention patent discloses the multiple oral Chinese medicine preparation that is used for the treatment of bladder cancer, discloses a kind ofly to be made into by five kinds of medicines such as Pseudobulbus Cremastrae seu Pleiones, Eriocheir sinensis, Herba Patriniae, Semen Euphorbiae, Herba Agrimoniaes the Chinese patent medicine of treatment bladder cancer as 01131500.8.01106271.1 disclose a kind of Chinese medicine for the treatment of cancer of forming by full shortcake, the Radix Euphorbiae Pekinensis, Herba Hedyotidis Diffusae, Herba Scutellariae Barbatae, Radix Et Rhizoma Rhei, Rhizoma Coptidis raw material.200610057952.2 disclosing a kind of is the Chinese medicine preparation of the treatment bladder cancer of feedstock production with Folium Pyrrosiae, the Radix Astragali, Herba Patriniae, Herba Solani Nigri, Pollen Typhae, Rhizoma Smilacis Glabrae, succinum, Herba Lysimachiae, Fructus Foeniculi, the Radix Linderae, Semen Plantaginis, Spora Lygodii, Herba Dianthi, the Rhizoma Atractylodis Macrocephalae and Radix Notoginseng.But still be difficult the irrigation of bladder medicine that Chinese medicine or Chinese medicine active ingredient extract or its compatibility compound recipe are formed in the prior art.

Summary of the invention

The invention provides and a kind ofly constitute, be used for the medicine of intravesical perfusion therapy bladder cancer with Chinese medicine active ingredient extract compatibility compound recipe,

The present invention is used for the treatment of the medicine of tumor of bladder, it is characterized in that medicine is made up of Chinese medicine monomer panaxoside Rg 3, Quercetin and curcumin, and the mol ratio of each component is ginsenoside Rg3: Quercetin: curcumin=20: 5: 3.

Optimal drug prescription of the present invention is three kinds of Chinese medicine monomer panaxoside Rgs, 3 160 ± 40 μ mol/L, curcumin 24 ± 6 μ mol/L, Quercetin 38 ± 9 μ mol/L.

The sweet temperature of Chinese medicine Radix Ginseng is gone into the heart, lung, spleen channel, strongly invigorating primordial QI, the mind calming and mentality promoting, invigorating the spleen and benefiting QI, nourishes heart and promotes the production of body fluid.The active component of Radix Ginseng antitumor action is the ginsenoside, studies show that the antitumor action of panaxoside Rg 3 is the most obvious, and the main mechanism of its antineoplastic is as follows: (1) promotes the apoptosis of tumor cell; (2) propagation of inhibition tumor cell; (3) suppressing tumor neogenetic blood vessels forms; (4) improve body's immunological function.

Quercetin is a kind of flavone compound, and the Chinese herbal medicine Flos Sophorae Immaturus, Cacumen Platycladi, Rhizoma Alpiniae Officinarum, Radix Notoginseng, Semen Ginkgo etc. all contain Quercetin.Previously studies confirm that Quercetin can suppress the growth of malignant tumor, numerous experimental studies results shows: Quercetin can suppress in tumor cell proliferation and the signal transduction pathway plurality of enzymes, remove effect such as oxygen-derived free radicals.Our Quercetin that studies confirm that can suppress transitional cell bladder carcinoma cell line propagation at present, and the apoptosis induction effect is arranged, and these effects may be worked by P53 and survivin albumen.Quercetin can reduce the methyl level of growth of bladder cancer cells suppressor gene Erb, P16 and PASSF1A simultaneously, referring to LiMa, JeanMagloire Feuguang, Jianzhong Lu, Shengiun Fu, Baoliang Ma, 1 BinqiangTian, Changping Zou ang Zhiping Wang.Growth inhibitory sffects of quercetin onbladder cancer cell.Frontiers in Bioscience 2006, September 1,11,2275-2285.

Rhizoma Curcumae Longae is the yellow natural plants that belongs to of Zingiberaceae, has stagnant, the diffusing wind of circulation of qi promoting to invigorate blood circulation and the pain relieving effect.Be not put into American Pharmacopeia in 2000.Present present the following aspects of anticancer mechanism table: (1) antioxidation to curcumin; (2) suppress the carcinogen activity, induce and separate toxenzyme; (3) formation of inhibition arachidonic acid metabolite; (4) suppress oncogene expression, apoptosis-induced, regulate intracellular signal transduction pathway.Curcumin can be induced the kinds of tumor cells apoptosis, can cause human liver cancer cell P53 and the significant change of C-myc albumen, referring to Jiang, M.C., H.f.Yang-Yen, J.K.Lin and J.J.Yen.Differential regulation ofP53, c-Myc, Bcl-2and Bax protein expression during apoptosis induced by widelydivergent stimuli in human hepatoblastoma cells.Oncogene, 1996,13 (3): 609-16..The inventor early-stage Study show that curcumin is significantly induced three kinds of tumor of bladder cell T24, EJ and UMU2 apoptosis, the retardance tumor cell in the G2/M phase.The down-regulated expression of two kinds of anti-apoptotic proteins Bcl-2 and Survivin, Bax and p53 up-regulated simultaneously.The clone forms test and confirms that curcumin heavy dose, shortterm effect have fatal effect in transitional cell bladder carcinoma cell line.The confirmation of in vivo test simultaneously curcumin is induced original position transitional cell bladder carcinoma cell line apoptosis, suppress and slow down the formation and development of bladder cancer, referring to Binqiang Tian, Zhiping Wang, YingmeiZhao, Degui Wang, Yonggang Li, Li Ma, Xiaoming Lia, Jing Lia, NanXIaoa, Yiaoa, Yichen Wanga, Ronald Rodriguezc.Effects of curcumin on bladdercancer cells and development of urothelia tumors in a rat bladder carcinogenesismodel.Canncer letters, 2008,8; 264 (2); 299-308..

The generation development of generally acknowledging at present tumor is the cumulative result of the rapid fellowship of polygenes, multistep, and this integral body sight with the traditional Chinese medical science has similarity.The present invention also follows this theoretical point view, see from integral body, adhere under Chinese medical theory instructs, with disease in conjunction with, side card is corresponding, principle-method-recipe-medicines is consistent is principle, carry out the effective monomer component compound compatibility, excavate the dabbling advantage Chinese medicine of tumor of bladder, from illustrating the anti-bladder cancer effect of compatibility compound recipe with tumor generation cells involved apoptosis.Ancient books " element is asked the opinion piece of writing of losing consciousness " is said: " the uterus transferring pathogenic heat to the urinary bladder, then infirmity hematuria ".The sick position of tumor of bladder is at bladder, and closely related with the heart, spleen, kidney, especially the most patients in operation back demonstrate,prove the marquis and show simulataneous insufficiency and excessive, and how based on blood stasis due to qi deficiency, so the Therapeutic Principle is when with benefiting QI for activating blood circulation, and warming and recuperating the spleen and kidney is advisable.Under these principles, determined the concrete medicine of prescription of the present invention, considered that simultaneously the present invention is a kind of medicine that is used for irrigation of bladder, so the present invention adopts by the active ingredient that extracts in the medicine.

Correlational study shows, the present invention acts on apoptosis and dead other groupings that obviously are better than comparing behind human bladder cancer's tumor cell, have antitumor, suppress the curative effect of tumor recurrence, enhancing immunity, and untoward reaction and the less advantage of side effect.

Description of drawings

Fig. 1 is the scatterplot of bivariate flow cytometer, is used for comparison the present invention and other grouping, acts on apoptosis and dead situation behind human bladder cancer's tumor cell.Fig. 2 acts on fluorescence microscope photo behind the tumor of bladder cell 24h for the present invention.

The specific embodiment

Most preferred embodiment of the present invention below is provided:

Chinese medicine active component compatibility complexity is loaded down with trivial details, and traditional research method is wasted time and energy, and also is not easy to find its inherent law.The orthogonal experiment design has crucial meaning as a kind of scientific method of multifactor multilevel experimental design in the research of medical science Chinese medicinal formulae.Most preferred embodiment of the present invention promptly adopts orthogonal experiment, draws best proportion compatibility, and concrete way is as follows:

1, cultivation and the cytotoxicity test of the external antitumor of three kinds of Chinese medicines test by the transitional cell bladder carcinoma cell line half-inhibition concentration (IC50) that calculates three kinds of Chinese medicine monomer.

" the half suppression ratio (IC50) of the three kinds of external anti-bladder cancer cell strain of monomer component T24 " obtain half suppression ratio (24h/IC50) result that three kinds of Chinese medicine monomer act on bladder cancer cell line T24 in 24 hours by experiment: panaxoside Rg 3 160 μ mol/L; Curcumin 24 μ mol/L; Quercetin 158 μ mol/L.

2, according to the Therapeutic Principle according to, be monarch drug with panaxoside Rg 3, Quercetin, curcumin are that the compound recipe ministerial drug is formed compound recipe.

3, use the mathematical statistics method, calculate the dose compatibility of the optimum medicine efficacy of three kinds of Chinese medicine monomer by the computer orthogonal design.L9 (34) orthogonal table is selected in this experiment for use, and the designed factor level table of this experiment is as follows:

Optimize the external anti-tumor of bladder test of compatibility compound recipe:

(1) Annexin V-FITC/PI detects apoptosis

The distribution of normal cell membrane phospholipid is asymmetric, and the film inner surface contains the phospholipid of negative electricity, and (as the acid of phosphatidyl silk, PS), and film outer surface contains and accounts for most neutral phospholipid.Apoptotic early stage, PS can be turned to the surface of cell membrane from the inboard of cell membrane, is exposed in the extracellular environment.Annexin V is that a kind of molecular weight is the Ca2+ dependency phospholipids incorporate albumen of 35～36kD, can combine with PS high-affinity specificity.Annexin V is carried out fluorescein FITC labelling can detect early apoptosis of cells.Propidium iodide (propidine io2dide, PI) be a kind of nucleic acid dye, it can not see through complete cell membrane, and the cell membrane of viable apoptotic cell is intact PI to be had achromatophilia, but at the cell and the dead cell of apoptosis middle and advanced stage, PI can permeate through cell membranes and is made that nucleus is red to be dyed.Therefore Annexin V is used with the PI coupling, just the cell and the dead cell in apoptosis late period morning can be made a distinction.Concrete experimental procedure slightly.

(2) cellulotoxic experiment obtains following orthogonal experiment tables of data:

Obtaining best compound compatibility dosage is: panaxoside Rg 3 160 μ mol/L, Quercetin 38 μ mol/L, curcumin 24 μ mol/L.

(3) Chinese medicine compound and monomer Chinese medicine and hydroxy camptothecin are to the contrast of human bladder cancer cell's strain T24 apoptotic effect.

Detect apoptosis through Annexin V-FITC/PI.Obtain the scatterplot of bivariate flow cytometer, see Fig. 1; The fluorescence microscope of experimental cell is seen Fig. 2 and Fig. 3.

Fig. 1 provides the scatterplot of bivariate flow cytometer, has compared compound Chinese medicinal preparation and other grouping, acts on apoptosis and dead situation behind human bladder cancer's tumor cell.Be the flow cytometry analysis result of Annexin V-FITC/PI detection apoptosis.Flow cytometer excitation wavelength 488nm is the passband filter detection FITC fluorescence of 515nm with a wavelength, and another wavelength detects PI greater than the filter of 560nm.The result judges: dyestuff such as PI that apoptotic cell is identified being useful on cytoactive have anti-metachromia, and non-viable non-apoptotic cell then can not.Cell membrane has the DNA of the cell of damage by PI to dye the generation red fluorescence, and the cell that cell membrane remains intact does not then have red fluorescence and produces.Therefore, apoptotic early stage PI not can dye and do not have the red fluorescence signal.Normal living cells similarly.On the scatterplot of bivariate flow cytometer, left lower quadrant shows living cells, is (FITC-/PI-); Right upper quadrant is non-living cells, and promptly non-viable non-apoptotic cell is (FITC+/PI+); And right lower quadrant is an apoptotic cell, manifests (FITC+/PI-).Cell after prescription effect of the present invention mainly is positioned at right upper quadrant and right lower quadrant, and the human bladder cancer's tumor cell after its effect is based on apoptosis and death, and living cells does not have substantially.Be better than other groupings by this experiment drug effect of the present invention as can be known.

The fluorescence microscope of experimental cell the results are shown in Figure 2 and Fig. 3.As seen from Figure 2, under fluorescence microscope, dichroic filter (FITC and rhodamine) is observed, and the bladder cancer tumor cell begins apoptosis after drug treating of the present invention, the bladder cancer tumor cell necrosis of having broken as seen from Figure 3.

Claims (3)

1, the medicine that is used for the treatment of tumor of bladder is characterized in that medicine is formed to be made of ginsenoside, Quercetin and curcumin, and the mol ratio of ginsenoside, Quercetin, three kinds of Chinese medicine monomer of curcumin is about 20: 5: 3.。

2,, it is characterized in that the dosage range of its best proportion compatibility is the ginsenoside: Quercetin: curcumin=160 ± 40 μ mol/L:24 ± 6 μ mol/L:38 ± 9 μ mol/L according to the medicine of the described treatment tumor of bladder of claim 1.

3, claim 1 or the 2 described medicines application in preparation treatment tumor of bladder medicine.

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