CN102526055B - Application of cyclohexanediamine hypocrelline B in photodynamic anti-tumor medicaments - Google Patents
Application of cyclohexanediamine hypocrelline B in photodynamic anti-tumor medicaments Download PDFInfo
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- CN102526055B CN102526055B CN201010606363.1A CN201010606363A CN102526055B CN 102526055 B CN102526055 B CN 102526055B CN 201010606363 A CN201010606363 A CN 201010606363A CN 102526055 B CN102526055 B CN 102526055B
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Abstract
The invention relates to an application of cyclohexanediamine hypocrelline B in photodynamic anti-tumor medicaments as a photosencitizer, and the photodynamic anti-tumor medicaments comprise the medicaments for treating cervical cancer, breast cancer, lung cancer, stomach cancer, skin cancer and other malignancies. The detection proves that the IC50 dark toxicity of the cyclohexanediamine hypocrelline B against HeLa, MCF7, MCF7/Adr, A549, A549/DDP, H1299, MGC803, A875 and other tumor cells when the cyclohexanediamine hypocrelline B is placed away from light is greater than 160 mu M; the IC50 phototoxicity of the cyclohexanediamine hypocrelline B against the detected tumor cells when the dose of red light is 12J/cm<2> for irradiation is less than 0.3 mu M; and a light enhancement factor (IC50 dark toxicity/IC50 phototoxicity) is greater than 500 and even greater than 1600. Mitochondria are further proven to be important action target points for photodynamic killing of the tumor cells through the cyclohexanediamine hypocrelline B, and the tumor cells mainly die in an apoptosis way.
Description
Technical field
The present invention relates to 2-demethoxy-2,3-ethylene-diamino hypocrellin B application in light power antitumor drug as photosensitizer.
Background technology
Optical dynamic therapy (photodynamic therapy, PDT) is associating light and photosensitizer, the emerging therapy that the body pathology tissue is treated.Optical dynamic therapy goes through and success in the U.S., Japan and other countries, for the clinical treatment of the malignant tumor such as the esophageal carcinoma, pulmonary carcinoma, oral cancer, hepatocarcinoma.Photosensitizer be a kind of can concentrate in pathogenic site and can the optical excitation at suitable wavelength under produce photochemical reaction and destroy the chemical substance of pathology target cell (or tissue).At present, development has the low and phototoxicity of dark toxicity to wait by force the novel photosensitive agent of premium properties is the key of field of photodynamic always.
Photofrin and hematoporphyrin derivative (Hematoporphyrin derivative, HpD) are first generation photosensitizer for photodynamic therapy medicines, are also the main optical dynamic therapy medicines of current clinical practice.The mixture that it is comprised of the porphyrin substance of different structure, have tissue selectivity and tumor cytotoxicity effect preferably.But their shortcoming is that chemical composition is not single, the skin phototoxicity time is long, requires patient to avoid illumination 4 to 6 weeks after injectable drug.In addition, it is at long wave direction absorption difference, and a little less than the penetrance of tissue, therefore not easy-to-use this photosensitizer of larger entity tumor is treated, and the laser of the 630nm used clinically can not be brought into play the maximum therapy effect.Therefore need the new photosensitizer of research and development to overcome these shortcomings.
Hypocrellin (Hypocrellin) is a kind of natural photosensitizer extracted in a kind of parasitical fungi Hypocrella bambusae (Bet Br). Sace (Hypocrellabambuas) from parasitizing the Yunnan Province of China Fargesia; belong in the awake analog derivative of perylene, be divided into hypocrellin (Hypocrellin A, HA) (see structural formula I) and HB Hypocrellin B (Hypocrellin B, HB) (seeing formula II).
With hematoporphyrin derivative, HpD compares, hypocrellin has that raw material is easy to get, photosensitizer triplet quantum yield and the creating singlet oxygen by using quantum yield is high, phototoxicity is high, dark toxicity is low, from the advantage such as the eliminating speed of normal structure is fast, its great advantage is easily to be fixed a point chemical modification, obtain highly purified monomer derived thing, it is a kind of second filial generation optical dynamic therapy medicine (Science Bulletin that has application prospect, 1990,35:1608-1616; Science Bulletin, 1990,35:1681-1691).But, as photosensitizer for photodynamic therapy, natural hypocrellin does not almost absorb at phototherapy window (600-900nm), has greatly limited its application clinically.In addition, natural hypocrellin is lipophilic compound, and water solublity is bad, is not easy to make medicament.Therefore, need to carry out structure of modification to hypocrellin, to obtain having at the phototherapy window photosensitizer for photodynamic therapy of strong absorption (being that red shift of wavelength is to coordinate the long wave laser therapy) and fat water compatible (being convenient to pharmaceutical preparation).
(English name is 2-demethoxy-2,3-ethylene-diamino hypocrellin B: 2-demethoxy-2,3-ethylene-diamino hypocrellin B, abbreviation EDAHB) (seeing formula II I) is a kind of derivant of HB Hypocrellin B, this derivant is that the prosposition at parent HB Hypocrellin B molecule adds hydrophilic group reacting ethylenediamine gained.
Chinese patent CN1600771A (open day: on March 30th, 2005) disclose preparation method and the application aspect optical function material and device thereof of 2-demethoxy-2,3-ethylene-diamino hypocrellin B; The people such as Xu Shangjie are at (Bioorganic& MedicinalChemistry Letters, 2004,14:1499-1501 and Photochemistry and Photobiology, 2004, reported optical physics and the spectrochemical property of 2-demethoxy-2,3-ethylene-diamino hypocrellin B in 80:112-114).But, using 2-demethoxy-2,3-ethylene-diamino hypocrellin B as photosensitizer, be applied at present there is not yet bibliographical information in light power antitumor drug.
Summary of the invention
The purpose of this invention is to provide 2-demethoxy-2,3-ethylene-diamino hypocrellin B application in light power antitumor drug as photosensitizer.
Tumor described in the present invention is breast carcinoma, cervical cancer, pulmonary carcinoma, gastric cancer or skin carcinoma.
The preferred adenocarcinoma of breast of breast carcinoma described in the present invention or doxorubicin resistant adenocarcinoma of breast.
The preferred adenocarcinoma of lung of pulmonary carcinoma described in the present invention, cisplatin resistance adenocarcinoma of lung or non-small cell adenocarcinoma of lung.
The preferred adenocarcinoma of stomach of gastric cancer described in the present invention.
The preferred melanoma of skin carcinoma described in the present invention.
The present invention has detected the light power antitumous effect of 2-demethoxy-2,3-ethylene-diamino hypocrellin B, and tumor cell used comprises the Several Kinds of Malignancy cell lines such as HeLa (cervical cancer tumer line), MCF7 (breast adenocarcinoma cell system), MCF7/Adr (amycin toleration breast adenocarcinoma cell system), A549 (lung adenocarcinoma cell system), A549/DDP (cisplatin toleration lung adenocarcinoma cell system), H1299 (non-small cell lung gland cell system), MGC803 (gastric adenocarcinoma cells system) and A875 (K-1735).Experimental result shows: 2-demethoxy-2,3-ethylene-diamino hypocrellin B when lucifuge to the IC50 of detected different tumor cells
dark toxicityall be greater than 160 μ M; And HONGGUANG dosage is 12J/cm
2during irradiation, to the IC50 of detected tumor cell
phototoxicityall be less than 0.3 μ M; Light enhancer (IC50
dark toxicity/ IC50
phototoxicity) all be greater than 500 and even be greater than 1600.In contrast, parent photosensitizer HB Hypocrellin B to the light enhancer of these tumor cells only between 10 to 30, far below the light enhancer of 2-demethoxy-2,3-ethylene-diamino hypocrellin B.
Test is discovery further, and 2-demethoxy-2,3-ethylene-diamino hypocrellin B is in optical dynamic therapy kill tumor cell processes, and the major way of death of neoplastic cells is apoptosis (apoptosis).Find that the mitochondrion of cell is subject to remarkable damage in 2-demethoxy-2,3-ethylene-diamino hypocrellin B light power inducing cancer cell death process simultaneously.This shows, mitochondrion is the important function target spot of 2-demethoxy-2,3-ethylene-diamino hypocrellin B light power inducing cancer cell death.
2-demethoxy-2,3-ethylene-diamino hypocrellin B of the present invention have composition single clear and definite, low to the dark toxicity of histiocyte while forming stable, lucifuge, light power anticancer effect is remarkable, anticancer spectrum is wide, action target spot is clear and definite and death of neoplastic cells be take apoptosis as the advantage such as main.The present invention is by detecting and find the external smooth power antitumous effect of ethylenediamine base HB Hypocrellin B, ethylenediamine base HB Hypocrellin B can be applied to optical dynamic treatment of tumor as photosensitizer, especially breast carcinoma, cervical cancer, pulmonary carcinoma, gastric cancer or skin carcinoma, and to the cancer of Chemoresistance as malignant tumor such as amycin toleration breast carcinoma, cisplatin toleration pulmonary carcinoma, thereby there is important application prospect.
2-demethoxy-2,3-ethylene-diamino hypocrellin B of the present invention, as active constituents of medicine, can be prepared into conventional pharmaceutical dosage form, for example, and injection etc.
Below by test example, further set forth antitumous effect of the present invention.
Test example 1:MTT method detects the action effect test of 2-demethoxy-2,3-ethylene-diamino hypocrellin B-optical dynamic therapy to tumor cell
(1) material
Cell line: HeLa, MCF7, MCF7/Adr, A549, A549/DDP, H1299, MGC803, A875 cell.
Reagent: RPMI 1640 culture medium and special top grade hyclone (U.S. GIBCO company); Dimethyl sulfoxide (DMSO), 3-(4,5-dimethylthiazole-2)-2,5-diphenyl tetrazole bromine salt (MTT) (U.S. Sigma company).
Equipment: KDH-150B red-light therapeutic instrument (Beijing Kedian Microwave Electronic Co., Ltd.); SPR-4001 spectroradiometer (Canadian Luzchem Research Inc.); Common inverted microscope (optical instrument factory, Chongqing); Biohazard Safety Equipment (U.S. BAKER company); Microplate reader (U.S. Bio Rad company).
(2) method
Cell culture: cell is at 37 ℃, saturated humidity, 5%CO
2cellar culture in incubator, culture fluid, for containing the two anti-RPMI-1640 of 10% calf serum, 1% penicillin and streptomycin, changes liquid and goes down to posterity in every 2 days.
Mtt assay is measured: by logarithmic (log) phase cell 0.25% trypsinization of cultivating, make single cell suspension, adjusting cell density is every milliliter 5 * 10
4individual, obtained cell suspension is inoculated on 96 well culture plates, and every hole 200 μ L, at 5%CO
237 ℃ of overnight incubation in incubator.The supernatant discarded culture fluid, add the photosensitizer working solution without the variable concentrations of phenol red RPMI-1640 dilution preparation with serum-free by experimental design under the condition of strict lucifuge.At 5%CO
2in incubator, 37 ℃ of lucifuges are hatched 1 hour.Supernatant discarded, phosphate buffer PBS washes twice.For dark toxicity test, every hole add 200 μ L containing 10% hyclone without phenol red RPMI-1640 culture medium, the continuation at 5%CO
2in incubator, 37 ℃ of lucifuges are cultivated 24 hours; For Phototoxicity experiment, every hole adds 200 μ L serum-frees without phenol red RPMI-1640, with the red light irradiation of various dose, recovers 10% hyclone then to the culture medium in every hole, continues at 5%CO
2in incubator, 37 ℃ of lucifuges are cultivated 24 hours.Discard subsequently culture fluid, every hole adds the MTT solution that the final concentration of the new preparation of 200 μ L is 0.5mg/mL, hatches 4 hours for 37 ℃.The careful suction abandoned liquid hole in, and every hole adds 150 μ L DMSO, vibrate 10min until blue crystallization dissolve fully.Select the 595nm wavelength, detect the absorbance value (A in each hole on microplate reader
595), do not add with parallel with experimental port the blank well zeroing that cell only adds MTT.
Cell survival rate=(test group A
595/ control group A
595) * 100%, wherein but matched group refers to and has absorbed photosensitizer the groups of cells of illumination not.
Light enhancer=IC50
dark toxicity/ IC50
phototoxicity, IC50 wherein
dark toxicityfor in dark toxicity test, (red light irradiation dosage is 0J/cm
2) cell survival rate corresponding photosensitizer concentration while being 50%; IC50
phototoxicityfor in Phototoxicity experiment, (HONGGUANG is 12J/cm according to dosage
2) cell survival rate corresponding photosensitizer concentration while being 50%.
Statistical method: adopt SPSS11.0 statistics software to carry out result treatment, continuous data adopts mean ± standard deviation
mean.
(3) result
This test example is to using parent photosensitizer HB Hypocrellin B as reference, has compared dark toxicity, phototoxicity and the light enhancer of 2-demethoxy-2,3-ethylene-diamino hypocrellin B and HB Hypocrellin B.
As shown in Figure 1, during not illumination, along with 2-demethoxy-2,3-ethylene-diamino hypocrellin B or HB Hypocrellin B are hatched the increase of concentration, the survival rate of HeLa cell all reduces gradually, shows that two kinds of photosensitizer have certain dark toxicity to cell.But, with HB Hypocrellin B, compare, the trend that 2-demethoxy-2,3-ethylene-diamino hypocrellin B causes cell survival rate to reduce is slower, for example, when 2-demethoxy-2,3-ethylene-diamino hypocrellin B concentration is increased to 160 μ M, cell survival rate still is about 80%, and HB Hypocrellin B concentration makes the HeLa cell survival rate just be down to 50% while being increased to 120 μ M.Table 1 and table 2 have further been listed respectively 2-demethoxy-2,3-ethylene-diamino hypocrellin B and the HB Hypocrellin B dark toxicity test result to the variety classes tumor cell.As shown in table 1, when 2-demethoxy-2,3-ethylene-diamino hypocrellin B concentration is increased to 160 μ M, the survival rate of all detected cells is about 80% left and right all, and this shows, for detected all tumor cell lines, the IC50 of 2-demethoxy-2,3-ethylene-diamino hypocrellin B
dark toxicityall be greater than 160 μ M; And as shown in table 2, for detected all tumor cell lines, the IC50 of HB Hypocrellin B concentration
dark toxicitybe about respectively: 120 μ M (HeLa), 45 μ M (MCF7), 60 μ M (MCF7/Adr), 55 μ M (A549), 110 μ M (549/DDP), 40 μ M (H1299), 60 μ M (MGC803), 90 μ M (A875).Therefore, the dark toxicity to tumor cell of 2-demethoxy-2,3-ethylene-diamino hypocrellin B is lower than parent photosensitizer HB Hypocrellin B.
As shown in Figures 2 and 3, during red light irradiation, the photosensitizer of low concentration just can make cell survival rate significantly reduce, and shows that two kinds of photosensitizer have higher phototoxicity to cell.But, with HB Hypocrellin B, to compare, more hurry up, for example, after HeLa cell and 0.2 μ M 2-demethoxy-2,3-ethylene-diamino hypocrellin B are hatched, give dosage is 12J/cm to the trend that 2-demethoxy-2,3-ethylene-diamino hypocrellin B causes cell survival rate to reduce
2red light irradiation, just can make cell survival rate be down to 50% (seeing Fig. 2); And, after HeLa cell and 5 μ M HB Hypocrellin B hatch, giving dosage is 12J/cm
2red light irradiation, make cell survival rate be down to 50% (seeing Fig. 3).Table 1 and table 2 have further been listed respectively 2-demethoxy-2,3-ethylene-diamino hypocrellin B and the HB Hypocrellin B phototoxicity measurement result to the variety classes tumor cell.As shown in table 1, all tumor cells for detected, give 12J/cm
2red light irradiation the time, the IC50 of 2-demethoxy-2,3-ethylene-diamino hypocrellin B
phototoxicitybe about respectively 0.2 μ M (HeLa), 0.1 μ M (MCF7), 0.1 μ M (MCF7/Adr), 0.2 μ M (A549), 0.2 μ M (549/DDP), 0.2 μ M (H1299), 0.2 μ M (MGC803), 0.3 μ M (A875); And as shown in table 2, all tumor cells for detected, give 12J/cm
2red light irradiation the time, the IC50 of HB Hypocrellin B
phototoxicitybe about respectively: 5 μ M (HeLa), 4 μ M (MCF7), 4 μ M (MCF7/Adr), 4 μ M (A549), 4 μ M (A549/DDP), 3 μ M (H1299), 3 μ M (MGC803), 5 μ M (A875).Therefore, the phototoxicity to tumor cell of 2-demethoxy-2,3-ethylene-diamino hypocrellin B is higher than parent photosensitizer HB Hypocrellin B.
IC50 according to photosensitizer to cell
dark toxicityand IC50
dark toxicityas a result, can calculate the light enhancer of two kinds of photosensitizer to different tumor cells.As shown in table 1, at HONGGUANG dosage, be 12J/cm
2during irradiation, 2-demethoxy-2,3-ethylene-diamino hypocrellin B all is greater than 500 to the light enhancer of tumor cell; And as shown in table 2, at HONGGUANG dosage, be 12J/cm
2during irradiation, HB Hypocrellin B is about respectively the light enhancer of tumor cell: 24 (HeLa), 11 (MCF7), 15 (MCF7/Adr), 14 (A549), 27.5 (A549/DDP), 13 (H1299), 20 (MGC803), 18 (A875).Therefore, the enhancer of the light to tumor cell of 2-demethoxy-2,3-ethylene-diamino hypocrellin B is higher than parent photosensitizer HB Hypocrellin B.
Dark toxicity, phototoxicity and the light enhancer measurement result of table 1 2-demethoxy-2,3-ethylene-diamino hypocrellin B to different tumor cells
Dark toxicity, phototoxicity and the light enhancer measurement result of table 2 HB Hypocrellin B to different tumor cells
The test of test example 2:DNA ladder band electrophoresis
(1) material
Cell line: HeLa and MCF7 cell.
Reagent: RPMI 1640 culture medium and special top grade hyclone (U.S. GIBCO company); DNA extraction agent box (Puli's lema gene technology company limited); Agarose (U.S. sigma company).
Equipment: KDH-150B red-light therapeutic instrument (Beijing Kedian Microwave Electronic Co., Ltd.); SPR-4001 spectroradiometer (Canadian Luzchem Research Inc); Common inverted microscope (optical instrument factory, Chongqing); Table-type high-speed refrigerated centrifuge (German HERAEMS company); Biohazard Safety Equipment (U.S. BAKER company).
(2) method
Cell culture: HeLa and MCF7 cell are at 37 ℃, saturated humidity, 5%CO
2cellar culture in incubator, culture fluid, for containing the two anti-RPMI-1640 of 10% calf serum, 1% penicillin and streptomycin, changes liquid and goes down to posterity in every 2 days.
The test procedure of DNA ladder band electrophoresis: will be in the HeLa of exponential phase cell and MCF7 cell, be divided into respectively three groups, under the condition of strict lucifuge, by experimental design, add respectively the photosensitizer working solution without 1 μ M of phenol red RPMI-1640 dilution preparation with serum-free.At 5%CO
2in incubator, 37 ℃ of lucifuges are hatched 1 hour.Then supernatant discarded, PBS washes twice, after adding serum-free without phenol red RPMI-1640, to three groups of every kind of cell, gives respectively 0J/cm
2, 12J/cm
2perhaps 18J/cm
2the red light irradiation of dosage, recover 10% hyclone in culture medium.Cell continues lucifuge and cultivates 24 hours.Collecting cell (2 * 10
6/ mL), PBS washes cell once.Extract the operating procedure of test kit according to apoptosis DNA Ladder and extract cell DNA.The DNA sample is at 1.5% agarose gel electrophoresis, and 12.5V/cm electrophoresis 1 hour, observe under uviol lamp and take pictures.
(3) result
After 2-demethoxy-2,3-ethylene-diamino hypocrellin B-optical dynamic therapy is processed cell, extract cell DNA, carry out agarose gel electrophoresis imaging, the result demonstration, the tumor cell that 2-demethoxy-2,3-ethylene-diamino hypocrellin B-optical dynamic therapy is processed has obvious DNA ladder zoning (seeing Fig. 4).Because the DNA ladder zoning is apoptotic characteristic indication, therefore, this result of the test shows that 2-demethoxy-2,3-ethylene-diamino hypocrellin B-optical dynamic therapy causes apoptosis of tumor cells.
Test example 3: mitochondrial membrane damage check test
(1) material
Cell line: HeLa cell.
Reagent: RPMI 1640 culture medium and special top grade hyclone (U.S. GIBCO company); Mitochondrion apoptosis detection kit (mitoCapt μ re
tMapoptosis detection kit) (U.S. Biovision company).
Equipment: KDH-150B red-light therapeutic instrument (Beijing Kedian Microwave Electronic Co., Ltd.); SPR-4001 spectroradiometer (Canadian Luzchem Research Inc); Common inverted microscope (optical instrument factory, Chongqing); Biohazard Safety Equipment (U.S. BAKER company); Laser confocal microscope (German Leica company).
(2) method
Cell culture adds in the special 35mm culture dish that posts coverslip in bottom, overnight incubation, during in exponential phase, hatch the 2-demethoxy-2,3-ethylene-diamino hypocrellin B working solution of cell and 1 μ M at cell 1 hour, after PBS washes three times, refusing illumination or giving dosage is 12J/cm
2red light irradiation, recover complete medium and continue lucifuge and cultivate 6 hours.Recycling test kit MitoCapt μ re
tMapoptosis Detection Kit labeled mitochondria, confocal laser scanning microscope.
(3) result
Cellular uptake 2-demethoxy-2,3-ethylene-diamino hypocrellin B, but not illumination, detect the cell mitochondrial of launching red fluorescence after 6 hours, its fluorescence is regular area distribution, and without diffusing phenomenon, the interior green fluorescence signal of Cytoplasm is weak (being shown in Fig. 5) very; After the cellular uptake 2-demethoxy-2,3-ethylene-diamino hypocrellin B, give 12J/cm
2red light irradiation, the cell mitochondrial of emission red fluorescence detected after 6 hours, but its fluorescence is dispersivity, distribute, the green fluorescence (seeing Fig. 6) that strong disperse shape distributes also detected in Cytoplasm simultaneously.Exist because dyestuff in test kit is easy to enter in Normocellular mitochondrion and with polymeric form, send bright red fluorescence; And, in apoptotic cell, because mitochondrial membrane potential is lost, mitochondrial membrane damages and makes permeability become large, dyestuff can't accumulate in mitochondrion, can remain in Cytoplasm with monomeric form, now sends green fluorescence.Therefore, can distinguish easily the cell mitochondrial of normal and damaged according to red green fluorescence.This result of the test shows, 2-demethoxy-2,3-ethylene-diamino hypocrellin B-optical dynamic therapy enlarges markedly the cell mitochondrial membrane permeability, and mitochondrial membrane is damaged.That is to say, mitochondrion is 2-demethoxy-2,3-ethylene-diamino hypocrellin B-important target spot of light power antineoplastic.
The accompanying drawing explanation
Fig. 1: 2-demethoxy-2,3-ethylene-diamino hypocrellin B and the HB Hypocrellin B dark toxicity figure to cell, wherein the dark toxicity icon of 2-demethoxy-2,3-ethylene-diamino hypocrellin B is ●, the dark toxicity icon of HB Hypocrellin B is ■;
Fig. 2: the phototoxicity figure of 2-demethoxy-2,3-ethylene-diamino hypocrellin B to cell;
Fig. 3: the phototoxicity figure of HB Hypocrellin B to cell;
Fig. 4: DNA ladder band electrophoretogram, wherein electrophoresis road 1 and 8 is the DNA molecular standard reference, the DNA that electrophoresis road 2,3 and 4 is the HeLa cell, the DNA that electrophoresis road 5,6 and 7 is the MCF7 cell, electrophoresis road 2 and 5 illumination dose are 0J/cm
2, electrophoresis road 3 and 6 illumination dose be 12J/cm
2, electrophoresis road 4 and 7 illumination dose be 18J/cm
2;
Fig. 5: the cell mitochondrial membrane permeability detects figure, and wherein 1 is Cytoplasm (very weak green fluorescence), and the 2nd, mitochondrion (red fluorescence);
Fig. 6: the cell mitochondrial membrane permeability detects figure, and wherein 1 is Cytoplasm (green fluorescence), and the 2nd, mitochondrion (red fluorescence).
The specific embodiment
Embodiment 1: the preparation of 2-demethoxy-2,3-ethylene-diamino hypocrellin B and purification
According to the disclosed method preparation of CN1600771A.Take raw material HB Hypocrellin B 200 grams, be dissolved in 200ml and newly steam in oxolane; Add 20 milliliters of ethylenediamines; 55 degrees centigrade of lucifuges stir 12 hours, and distilling under reduced pressure is except desolventizing; With 200 milliliters of chloroform dissolution precipitations, extremely neutral with 1% dilute hydrochloric acid adjust pH; Boil off chloroform; By 200 gram re-crystallizing in ethyl acetate; Then purify residue by thin layer chromatography, obtain 2-demethoxy-2,3-ethylene-diamino hypocrellin B, productive rate is 55%.Measure the purity of 2-demethoxy-2,3-ethylene-diamino hypocrellin B with high speed liquid chromatography, purity is higher than 95%.
2-demethoxy-2,3-ethylene-diamino hypocrellin B obtained above is preserved with lyophilized form, during use, with DMSO, dissolved and be made into storage liquid, by PBS or culture medium, be diluted to the desired concn working solution.
Embodiment 2: the preparation of injection
Get the lyophilized powder 1g of the 2-demethoxy-2,3-ethylene-diamino hypocrellin B of embodiment 1 preparation, adopt liposome, add pH adjusting agent, osmotic pressure regulator, be prepared into injection.
Claims (2)
1. 2-demethoxy-2,3-ethylene-diamino hypocrellin B application in preparing light power antitumor drug as photosensitizer, it is characterized in that: described tumor is breast carcinoma or pulmonary carcinoma.
2. application according to claim 1 is characterized in that: described breast carcinoma is adenocarcinoma of breast or doxorubicin resistant adenocarcinoma of breast.
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| Title |
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| 竹红菌素及其衍生物抗肿瘤活性的研究进展;肖彩霞等;《食品与药品》;20081231;第10卷(第7期);55-59 * |
| 肖彩霞等.竹红菌素及其衍生物抗肿瘤活性的研究进展.《食品与药品》.2008,第10卷(第7期),55-59. |
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