CN1442134A - Application of scutellaria root extract in preparation of anti esophageal cancer medicine - Google Patents
Application of scutellaria root extract in preparation of anti esophageal cancer medicine Download PDFInfo
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- CN1442134A CN1442134A CN 03109933 CN03109933A CN1442134A CN 1442134 A CN1442134 A CN 1442134A CN 03109933 CN03109933 CN 03109933 CN 03109933 A CN03109933 A CN 03109933A CN 1442134 A CN1442134 A CN 1442134A
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Abstract
An application of the scutellaria extract in preparing the medicine for esophagus cancer is disclosed, and it features that scutellarin, scutellaroside, or their mixture can be used to prepare the pharmacologically receptable sale for suppressing the splitting of cancer cell, inducing the withering of cancer cell and inhibiting the generation of the blood vessel inside esophagus cancer.
Description
Technical field
The present invention relates to the application in the anti-esophageal carcinoma medicine of preparation of the purposes of Radix Scutellariae extract, particularly Radix Scutellariae extract.
Technical background
Generally investigate from the long-term case of esophageal carcinoma, the evolution of esophageal cancer cell is to begin progressively to develop into the severe hypertrophy from slight hypertrophy, cancer in situ and infiltrating carcinoma, development is slower from the slight hypertrophy of mucous membrane of esophagus to cancer in situ, have and think about 5 to 10 years of this evolution process, in case form infiltrating carcinoma, its development speed is quite fast, block symptom when patient has obviously to swallow, cure rate around here is very low.The treatment of esophageal carcinoma is at present still based on operation and radiotherapy, because chemotherapy is bigger to the toxicity of human body, only as a kind of supplementary means.And recur again in a short time easily and spread, and there are a lot of patients to refuse thus to be treated surgically at present operative treatment.In radiotherapy, in case the patient has dyscrasia, perforation, CT to confirm have serious trachea or trunk to be invaded or serious chest and back pain, all radiotherapies again such as leukocyte rising, taboo relatively.
The radiotherapy of esophageal carcinoma is different from the oncotherapy at other position, since tumor all the time with esophagus in one, when the planning target area, tube chamber and pathological changes are put under together, if the same with the tumor at other position of irradiation, a high-dose irradiation will certainly cause the damage of mucous membrane of esophagus, even perforation, discovery being arranged when the directional emittance of lung tumors, esophagus is accepted the 24-30Gy amount, radiation injury can occur, later stage performance esophagus locality is narrow.Again since the esophageal carcinoma patient during with regard to disease the overwhelming majority belonged to late period, not only local patholoic change is extensively soaked into, and send out or lymphatic metastasis the distant place of existing subclinical focus, postmortem is found, manyly clinically think when local early stage case is dead, existing companion's metastasis more than half, there is widely lymph node be invaded more than 70%, the case less than 1/3 of real energy radical excision, 5 years survival rates of conventional radiotheraphy only are 13%, and local recurrence also is the main cause of esophageal carcinoma operation and radiotherapy failure, accounts for more than 80%, this shows that it is far from being enough that the treatment of esophageal carcinoma only depends on operation and radiotherapy.
Because in chemotherapy process, with side reaction greatly and complication, even by the radiotherapy combined chemotherapy, its 5 years survival rates also only about 13%.Existence person more than 5 years also was greatly affected for quality of life in later stage, can not fundamentally treat esophageal carcinoma effectively, more can not accomplish to prevent trouble before it happens.
Baikal skullcap root (Scutellaria baicalensis Georgi) head is stated from Shennong's Herbal, has another name called Huang Wen, no a kind of reed mentioned in ancient books etc.According to the report of existing document, the pharmacological effect bitter in the mouth of Radix Scutellariae, cold in nature, function is let out excess-fire, except that damp and hot, detoxifcation, hemostasis, antiabortive, various bacteria all there is inhibitory action, to influenza virus, dermatophytes also has inhibitory action, and heat extraction is arranged, detoxifcation, calmness, blood pressure lowering, function of gallbladder promoting, smooth muscle spasm is removed in diuresis, suppresses effects such as intestinal tube wriggling.The acetone extract of Radix Scutellariae also is expected to become the Natural antioxidant of preserving processed food.Root, the leaf extract of Radix Scutellariae have antiulcer activity.
Baicalin in the Radix Scutellariae extract and baicalin, existing its pharmacological action of bibliographical information is antiinflammatory, anti-allergy action; The hepatic cholagogic effect; Blood pressure lowering, diuresis, calmness, refrigeration function; The aldose reductase inhibitory action.Clinical practice is mainly used in infectious hepatitis, acute biliary infection, lead poisoning, heat-clearing and toxic substances removing, vomiting and nausea uncomfortable in chest, cough due to lung-heat, frequent fetal movement and sore etc.With regard to baicalin, the in vitro study report that is confined at breast cancer cell is arranged also.But these researchs all also are in very early stage, and are also fuzzy for the antitumaous effect mechanism of Radix Scutellariae extract.
Summary of the invention
The objective of the invention is provides a kind of preparation to have efficiently for the deficiency that solves above-mentioned technology, uses Radix Scutellariae extract in the anti-esophageal carcinoma medicine that has no side effect, i.e. the application of Radix Scutellariae extract in the anti-esophageal carcinoma medicine of preparation.
In order to achieve the above object, the invention provides a kind of Radix Scutellariae extract, use the Radix Scutellariae extract drug prepared and be used for the treatment of or prevent esophageal carcinoma in the application of preparation in the anti-esophageal carcinoma medicine.Radix Scutellariae extract refers in particular to the extract with flavone precursor structure, can be baicalin, also can be baicalin, can also be both mixture, and both proportionings can be that any proportioning is mixed in the mixture.Described Radix Scutellariae extract can be the pharmaceutically acceptable salt that Radix Scutellariae extract is made, as with following acid: similar acid-addition salts such as maleic acid, fumaric acid, succinic acid, tartaric acid, acetic acid, hydrochloric acid, hydrobromic acid, sulphuric acid, phosphoric acid; Can add suitable solvent when needing, as analog such as water, methanol, ethanol, ether, oxolanes.Radix Scutellariae extract and pharmaceutically acceptable salt can oral, drug administration by injection, as powder, granule, tablet, piller, capsule, Emulsion, syrup, injection etc.Described preparation can prepare with conventional method.Described Radix Scutellariae extract with flavone precursor structure, its flavone precursor structure is as shown below:
Described baicalin is baicaligenin again, and flavone compound is one of main active of Radix Scutellariae.Molecular weight 270.25 can be obtained by the baicalin hydrolysis.
The structure of baicalin: on the flavone precursor structure 4 ', 5,6,7 four hydrogen base-H are replaced by hydroxyl-OH.
The structure of baicalin is: on the flavone precursor structure 4 ', 5,6 three hydrogen base-H are replaced 7 hydroxyl-OH and glucuronic acid condensation, i.e. baicalin-7-O-glucuronic acid by hydroxyl-OH.
The application of Radix Scutellariae extract of the present invention in preparation esophageal carcinoma medicine discloses the new purposes of Radix Scutellariae extract in pharmacy.Radix Scutellariae extract, particularly baicalin have tangible inhibitory or killing effect to esophageal cancer cell.
Apoptosis is to be prevalent in intravital a kind of spontaneous, the process of cell death initiatively of cell biological, be that organism is kept the metastable intrinsic mechanism of cell quantity, this mechanism is if having obstacle or take place unusual, just might cause tumor or other pathological changes, tumor is that a kind of apoptosis is crossed slow and bred too much disease, if can suppress the propagation of tumor cell and induce its apoptosis, just can control the generation and the development of tumor effectively.In addition, when diameter of tumor during in the 1-2 mm square, because tumor body central part anoxia, tumor cell can produce angiogenesis factor, like this, tumor can obtain required nutrition of its spontaneous growth and oxygen, by to this neovascularity inhibition of proliferation, help to suppress the continued growth of tumor cell, therefore, suppress the growth of tumor cell, need to seek and to suppress the propagation of tumor cell and induce its apoptosis and have the material of angiogenesis inhibiting activity.
And of the present invention discovering, baicalin in the Radix Scutellariae extract, its special mechanism of action are directly to suppress the division and the hypertrophy of esophageal cancer cell, and have the effect of induced strong cancer cell-apoptosis, and normal cell growth cycle and apoptosis are not had obvious effect; Another mainly acts on is to suppress the esophageal carcinoma internal blood vessel to generate, particularly under anaerobic environment revascularization, thereby the blood supply of cutting off tumor effectively makes its degeneration necrosis.Particularly in the isolated cells culture experiment, show as the inhibition of esophageal cancer cell release angiogenesis factor and the apoptosis of induction of vascular endothelial cell.In addition, in living animal (nude mice) experiment, discovery can press down the growth of killing transplantation tumor significantly, the highlyest effectively reaches 95%, and without any side effects.
Radix Scutellariae extract has been excavated in the application of Radix Scutellariae extract of the present invention in pharmacy, particularly baicalin and the baicalin new purposes in the production field of esophageal carcinoma treatment and prophylactic agent.The drug toxicity of Radix Scutellariae extract of the present invention, particularly baicalin, baicalin preparation is low, and human body is had no side effect.Radix Scutellariae extract of the present invention, particularly baicalin, baicalin raw material sources are extensive, and cost is lower.The present invention is for the esophageal carcinoma treatment and prevent capturing early of this difficult problem to have great realistic significance.
In order to understand essence of the present invention better, the present invention is further described below in conjunction with contrast pharmacological evaluation and result.Experimental result 1:
As shown in Figure 1, this picture is baicalin (baicalein) the inducing cancer cell karyorhexis in the Radix Scutellariae extract, and apoptosis starts from nuclear cracking.Show among the figure that tangible nucleus cracking (arrow indication among the right figure) appears in esophageal cancer cell after baicalin is handled 2 days.Baicalin does not have this effect (left figure) to normal cell.DAPI immunostaining (special cells nuclear staining method) is adopted in this experiment.Control is undressed cellular control unit.Experimental result 2:
As shown in Figure 2, this photo is the apoptosis situation that baicalin is induced esophageal cancer cell.Special immunohistochemical staining method (tunel Stain) is adopted in this experiment.The method is the optimum cell apoptosis identification of means of generally acknowledging in the present world wide, apoptosis or just after apoptotic cells is dyed, be displaing yellow or crocus, and normal cell then manifests redness.Photo demonstration esophageal cancer cell has been induced the apoptosis of these cells after baicalin (baicalein) is handled 3 days, then unaffected through the same normal cell of handling.Experimental result 3
As shown in Figure 3, this photo is the apoptosis situation of baicalin induction of vascular endothelial cell.Special immunohistochemical staining method (tunel stain) is adopted in this experiment. and apoptosis or just presenting yellow after apoptotic cells is dyed, normal cell then presents redness.Shown two kinds of blood vessel endotheliums in the picture, promptly criticized chrotoplast (HAEC) in Human umbilical vein endothelial cells (HUVEC) and the human artery, induced the apoptosis of these cells after baicalin (baicalein) is handled 3 days, control is undressed cellular control unit.Experimental result 4
As shown in Figure 4, this table is that baicalin suppresses the experimental status that esophageal cancer cell discharges angiogenesis factor.
Experimental technique: 1 * 10
5Esophageal cancer cell is planted on the culture plate that diameter is 15mm, and culture fluid contains 10% calf serum.After treating cell attachment in 24 hours, change culture fluid into serum-free medium.Change beginning dosing in 24 hours behind the liquid.After different time is handled, culture fluid is collected, with the content of ELISA mensuration culture fluid medium vessels somatomedin, measured content needs to proofread and correct through actual cell number.Esophageal cancer cell is after baicalin is handled 2 days, and angiogenesis factor discharges obviously and reduces.(1%O in anoxic environment
2), the baicalin inhibitory action is especially remarkable.Experimental result 5
As shown in Figure 5, this photo shows that baicalin suppresses nude mice and goes up the angiogenesis situation of transplanting in the esophageal carcinoma.
Experimental technique: it is subcutaneous earlier the esophageal cancer cell strain to be seeded in nude mice.From the 2nd week beginning, baicalin is through subcutaneous administration, 3 times weekly.In 12 weeks of Therapy lasted to the, measure the surface area of a tumor weekly.Experiment separates tumor after finishing on nude mice, after chemical fixation and frozen section, carry out immunohistochemical staining with eight factors as antibody.Brown fragment is the tumor vessel section in the picture.Left figure sample is taken from not medication group, and right figure sample is taken from the medication group, dosage 1mg/kg body weight, and visible vessels quantity obviously reduces.Experimental result 6:
As shown in Figure 6, this table is the DNA division situation that baicalin (baicalein) suppresses esophageal cancer cell.Visible esophageal cancer cell is after baicalin is handled among the figure, and the cell number that is in DNA synthesis stage (S phase) obviously reduces, the presynthetic phase (G0/G1) cell number then obviously increase.Since these cells the presynthetic phase G0/G1 accumulation, thereby suppressed this proliferation of cells effectively.Experimental technique: esophageal cancer cell is after baicalin is handled, again through fluorescent labeling, with the cell cycle analysis method cell number that is in each different division stages is carried out quantitative assay at last, the ratio that measured numerical value is accounted for total cell number compares and obtains chart.Experimental result 7
As shown in Figure 7, this chart is that baicalin (baicalein), baicalin (baicalin) and matched group usefulness Semen Coicis extract (wogonin) press down the curative effect comparison of killing esophageal cancer cell.In stripped sex culture experiment, but baicalin, baicalin and Semen Coicis extract all have the effect of esophageal cancer cell extremely, but action intensity is slightly variant.Relative intensity: baicalin＞baicalin＞Semen Coicis extract.The result tests 8
As Fig. 8, shown in Figure 9, chart is that baicalin (baicalein) has suppressed the expression (Fig. 8) of Cyclooxygenase (COX-2) and synthetic (Fig. 9) of prostaglandin.COX-2 is the enzyme that a kind of catalysis prostaglandin generates, and this enzyme and its product prostaglandin have played important effect in the generation of tumor and development.One of baicalin main mechanism is to play the effect of cancer inhibitting and killing cell by its these inhibitory action.Experimental technique: Fig. 8 adopts West Blot Analysia method to measure COX-2 protein expression level, and the right numeral is pointed out the proteinic molecular weight of COX-2.Fig. 9 adopts the ELISA method to measure by esophageal cancer cell and is released into content of prostaglandin in the culture fluid, and does to take statistics after the alignment with actual cell number and learn processing.Experimental result 9:
As shown in figure 10, this table is that baicalin presses down the dose curve that kills esophagus growth of cancer cells in the cell culture experiments in vitro.Curve 1 expression normal cell, curve 2 expression breast cancer cells, curve 3 expression esophageal cancer cells.
Experimental technique: identical condition of culture is all adopted in three kinds of experimental cell strains.1 * 10
5Cell is planted on the culture plate that diameter is 15mm, and culture fluid contains 10% calf serum.After treating cell attachment in 24 hours, change culture fluid the influence of serum-free medium into the somatomedin cell growth avoiding existing in the serum.Change beginning dosing in 24 hours behind the liquid.Dosing was made cell counting after 5 days.Before the counting earlier with Tai Panlan with cell dyeing to get rid of the influence of cell of dying in heaven to experimental result.Every bit is the meansigma methods of 3 experiments in the curve.
As seen, be 0.1 to have begun effect (20% suppress) with concentration from curve, when concentration was 0.5 μ g/ml, the inhibition of esophageal cancer cell growth was 50%.And when concentration reached 10 μ g/ml, inhibition can reach 95%.Baicalin does not have obvious effect to normal cell.As seen baicalin can press down the growth of the esophageal cancer cell that kills In vitro culture effectively.Experimental result 10
As Figure 11, shown in Figure 12, be that baicalin presses down the growth of killing transplanting esophageal carcinoma on the living animal.Experimental technique: it is subcutaneous earlier the esophageal cancer cell strain to be seeded in nude mice, through 1-2 after week tumor begin growth.The baicalin subcutaneous injection is from the beginning of second week, 3 times weekly.Laboratory animal is divided into matched group of 3 test dose combinations, every group of 10 nude mices at random.In 12 weeks of Therapy lasted to the, measure the surface area of an esophageal carcinoma tumor weekly.Experiment separates the esophageal carcinoma tumor after finishing on nude mice, and does to preserve to handle and prepare against as pharmacology, SABC and biochemical analysis.Figure 11 shows: baicalin presses down the metering curve that kills transplantation tumor.Figure 12 shows: but baicalin has obviously transplantation tumor kills effect.Experimental result 11:
When following experiment is independent medication of baicalin (baicalein) and baicalin (baicalin) or mixed reagent the esophageal cancer cell of isolated culture and the effect of breast cancer cell are compared.
Experimental technique: 1 * 10
5Esophageal cancer cell is planted on the culture plate that diameter is 15mm, and culture fluid contains 10% calf serum.Treat (about 24 hours) behind the cell attachment, change culture fluid the influence of serum-free medium into the somatomedin cell growth avoiding existing in the serum.Change beginning dosing in 24 hours behind the liquid.Dosing was made cell counting after 5 days.Before the counting earlier with Tai Panlan with cell dyeing to get rid of the influence of cell of dying in heaven to experimental result.As every bit among Figure 13 is the meansigma methods of three experiments.Experimental result:
As Figure 13, shown in Figure 14, separately during medication, the action intensity of baicalin is higher than baicalin.When baicalin and baicalin mixed reagent (1: 1), it has slight increase to the inhibitory action of esophageal cancer cell growth when the low dose, but after dosage increases, and its effect is greater than baicalin (medication separately), but less than baicalin (medication separately).Matched group has increased the growth of the esophageal cancer cell of isolated culture, does not have any inhibitory action.Show as Figure 14, when baicalin (baicalein) and baicalin (baicalin) and mixed reagent thereof to the effect of the esophageal cancer cell and the breast cancer cell of isolated culture.
Above-mentioned experimental result Radix Scutellariae extract as can be seen is used to prepare anti-esophageal carcinoma medicine, can obviously suppress the division of esophageal cancer cell, and has the effect of induced strong cancer cell-apoptosis, and normal cell growth cycle and apoptosis are not had obvious effect; And tool inhibition esophageal carcinoma internal blood vessel generation, the particularly revascularization under anaerobic environment, thereby the blood supply of cutting off tumor effectively makes its degeneration necrosis.Particularly the baicalin medicine that is used to prepare treatment or prevention esophageal carcinoma has very active operation significance.
Description of drawings
Fig. 1 is that baicalin (baicalein) is induced esophageal cancer cell karyorhexis experiment photo;
Fig. 2 is the apoptosis situation experiment photo that baicalin is induced esophageal cancer cell;
Fig. 3 is the apoptosis situation experiment photo of baicalin induction of vascular endothelial cell;
Fig. 4 is that baicalin suppresses the experimental status photo that esophageal cancer cell discharges angiogenesis factor;
Fig. 5 is that baicalin suppresses the upward interior angiogenesis situation of transplanting esophageal carcinoma of nude mice;
Fig. 6 is the DNA division situation that baicalin (baicalein) suppresses esophageal cancer cell;
Fig. 7 presses down the curative effect of killing esophageal cancer cell to compare;
Fig. 8 is that baicalin (baicalein) has suppressed the expression of Cyclooxygenase (COX-2);
Fig. 9 is the synthetic situation that baicalin (baicalein) has suppressed prostaglandin;
Figure 10 is that baicalin presses down the growth of killing transplanting esophageal carcinoma on the living animal;
Figure 11 is that baicalin presses down the metering curve that kills transplantation tumor;
Figure 12 is that baicalin kills effect to pressing down of transplantation tumor;
Figure 13 be baicalin (baicalein) and baicalin (baicalin) separately when medication or mixed reagent to the effect of the esophageal cancer cell of isolated culture relatively;
Figure 14 is that Radix Scutellariae extract compares esophageal cancer cell and mastocarcinoma cytosis.
The specific embodiment
Present embodiment provides the extracting method of Radix Scutellariae extract, and at first decocting boils Radix Scutellariae coarse powder twice, adds acid and transfers to PH1-2, and the 70-80 insulation was left standstill 12-24 hour, filtered.Precipitate adds the water stirring makes into suspension, transfers PH7-7.5 to dissolving with alkali, adds ethanol and stirs, filtration.Filtrate is the Radix Scutellariae extracting solution with effective Radix Scutellariae extract composition.
Embodiment 2:
Present embodiment provides the medicine that utilizes the existing goods baicalin to be used to prepare anti-esophageal carcinoma, takes by weighing baicalin (commercially available) 300 grams, mannitol 75 grams, sodium citrate 250 grams, powder pin preparation technology with routine makes 1000 powder pins, and every contains baicalin 300mg, as anti-esophageal carcinoma medicine.
Embodiment 3:
Present embodiment provides and will obtain baicalin after the baicalin hydrolysis, takes by weighing 100 gram baicalin raw materials,
Add lactose 66g, pregelatinized Starch 64g, microcrystalline Cellulose 66g, magnesium stearate 3g, film-making technology routinely makes 1000 in tablet, is equivalent to every and contains 100 milligrams of baicalins, as anti-esophageal carcinoma medicine.
Embodiment 4:
Present embodiment provides baicalin and baicalin 1: 1 mixing by weight.Take by weighing 100 gram mixed materials, add lactose 66g, pregelatinized Starch 64g, microcrystalline Cellulose 66g, magnesium stearate 3g makes 1000 capsules with the capsules preparation technique of routine, and every capsules contains baicalin and baicalin mixture 100mg, as anti-esophageal carcinoma medicine.
Claims (6)
1, the application of Radix Scutellariae extract in the anti-esophageal carcinoma medicine of preparation.
2, the application of Radix Scutellariae extract according to claim 1 in the anti-esophageal carcinoma medicine of preparation is characterized in that described Radix Scutellariae extract has the flavonoid precursor structure, and described precursor structure is as follows:
3, the application of Radix Scutellariae extract according to claim 1 in the anti-esophageal carcinoma medicine of preparation is characterized in that described Radix Scutellariae extract is a baicalin.
4, the application of Radix Scutellariae extract according to claim 1 in the anti-esophageal carcinoma medicine of preparation is characterized in that described Radix Scutellariae extract is a baicalin.
5, the application of Radix Scutellariae extract according to claim 1 in the anti-esophageal carcinoma medicine of preparation is characterized in that described Radix Scutellariae extract is the mixture of baicalin and baicalin.
6, the application in the anti-esophageal carcinoma medicine of preparation according to claim 1 or 2 or 3 or 4 or 5 described Radix Scutellariae extracts is characterized in that described Radix Scutellariae extract is the pharmaceutically acceptable salt that Radix Scutellariae extract is made.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2006081740A1 (en) * | 2005-02-01 | 2006-08-10 | Shanghai Gloriayx Biopharmaceuticals Co., Ltd | The synergistically pharmaceutical composition of baicalein and baicalin for inhibiting tumor |
| CN1879615B (en) * | 2005-01-20 | 2010-05-26 | 上海格鲁奥丽生物医药技术有限公司 | Anti-tumor synergistic pharmaceutical composition |
| WO2021000558A1 (en) * | 2019-07-04 | 2021-01-07 | 兰州大学 | Activating agent for simultaneously activating oxidative phosphorylation pathway and inhibiting glycolytic pathway and application thereof |
-
2003
- 2003-04-08 CN CNB031099335A patent/CN100446761C/en not_active Expired - Fee Related
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1879615B (en) * | 2005-01-20 | 2010-05-26 | 上海格鲁奥丽生物医药技术有限公司 | Anti-tumor synergistic pharmaceutical composition |
| WO2006081740A1 (en) * | 2005-02-01 | 2006-08-10 | Shanghai Gloriayx Biopharmaceuticals Co., Ltd | The synergistically pharmaceutical composition of baicalein and baicalin for inhibiting tumor |
| JP2008528640A (en) * | 2005-02-01 | 2008-07-31 | シャンハイ グルアォリ バイオファーマシューティカルズ カンパニー リミテッド | Antitumor synergistic pharmaceutical composition of baicalein and baicalin |
| WO2021000558A1 (en) * | 2019-07-04 | 2021-01-07 | 兰州大学 | Activating agent for simultaneously activating oxidative phosphorylation pathway and inhibiting glycolytic pathway and application thereof |
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Assignee: Hangzhou Zhuyangxin Pharmaceutical Co., Ltd. Assignor: Hangzhou Huadong Medicine Group Biological Engineering Research Institute Co., Ltd. Contract record no.: 2011330000095 Denomination of invention: Application of scutellaria root extract in preparation of anti esophageal cancer medicine Granted publication date: 20081231 License type: Exclusive License Open date: 20030917 Record date: 20110314 |
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