CN1699369A - Podophyllotoxin compounds and their application and preparation process - Google Patents
Podophyllotoxin compounds and their application and preparation process Download PDFInfo
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Abstract
The invention relates to novel podophyllotoxins compound the use of the compound and the process for preparing the compound.
Description
Technical field
The present invention relates to the new podophyllotoxin analogue of a class, and the preparation method of the purposes of this compound and this compound.
Background technology
Podophyllotoxin (Podophyllotoxin) is that a class is mainly separated the remarkable Cytotoxic natural active matter that has that obtains from the Berberidaceae may apple, the medicinal plant that very long applicating history is arranged again, for a long time, the Podophyllum emodi var chinense lignans is widely used in blood circulation promoting and dispersing pathogen accumulation as medicinal ingredients always, the worm snakebite, pain furuncle poison, fall and beat, bones and muscles pain due to rheumatism, emetic, symptoms such as wart knurl and bronchitis. from nineteen forties, King and Shliivan reported first podophyllotoxin have effect (the Stahelin H.F. of similar colchicine, Eur.J.Cancer, 1970,6,303), but, once in for some time, limited its application clinically because of its toxic side effect.Afterwards, many chemists and medicine scholar modify its structure, in the hope of obtaining the podophyllotoxin medicine that anti-tumor activity is good and toxic side effect is little.Twentieth century six the seventies, mountain pass scholar company finds 4 in succession '-β-D-glucose-derivative VP-16 (etopside) and the VM-26 following products such as (teniposide) of Demethylepipodophyllotoxin fat element have the activity that intensive suppresses tumour cell,
Nineteen eighty-three VP-16 is by U.S. FDA official approval listing (New York:Academic Press Inc., 1984).
The main synthetic route of VP-16 can be referring to 1) Beijing Pharmaceutical Ind. Inst. one Room, Beijing medicine industry, 2:11,1984; 2) day disclosure special permission communique, 61-134396; 3) Chinese patent CN 1057054A; WO 93/02094; 4) Chinese patent CN 1337402A).Etopophos (Brit 1989:2207674) is 4 of VP-16 '-organic phosphate disodium salt, goes on the market in the U.S. in 1996.Also has NK-611 (day disclosure special permission 88-10789 in addition; Drug Fut 1991,16 (2), 113), (Drug Fut 1996,21,1136 for TOP-53; CancerRes 1996,56,2809), GL-331 several medicines such as (Mol Pharmacol 1996,49,721) have entered the clinical II phase or the III phase tests, will go on the market soon.Test shows, podophyllotoxin medicines such as VP-16 are to small cell lung cancer, the Fei Hejieshi disease, acute monocytic leukemia, the property satisfied monocytic leukemia, mammary cancer, bladder cancer, numerous tumour cells such as carcinoma of testis have better therapeutic effect, but it is strong to exist resistance, poorly water-soluble, shortcomings such as serious bone marrow depression and oral result difference, seek the new about thing of Podophyllum emodi var chinense class that anti-tumor activity is good and toxic side effect is little, remain a big focus of pharmacy circle.
Summary of the invention
The object of the present invention is to provide the novel Podophyllum emodi var chinense class medicine antitumor cell compound of a class, the compound expection that this class is new has good water solubility, and lower toxicity and higher antitumor action are arranged.The invention provides purposes of this class new compound and preparation method thereof simultaneously.
Compound molecule formula of the present invention is formula as follows:
R can be a hydrogen in the formula, or methyl, or sec.-propyl, or first sulphur methylene radical, or new butyl, or isobutyl-, or benzyl, or methylol, or the 2-hydroxyethyl, or to hydroxybenzyl.
Compounds process for production thereof of the present invention is with podophyllotoxin and N-(1-oxygen base-2; 2,6,6-tetramethyl--oxygen-carbonyl)-L-amino acid is dissolved in the exsiccant methylene dichloride; under nitrogen protection, stir; add dicyclohexylcarbodiimide (DCC) then, stirring reaction under nitrogen protection refilters after question response is finished and removes white precipitate; after removing solvent under reduced pressure; crude product through column chromatography purification with volume ratio is: 15: 1 methylene dichloride-acetone wash-out gets product.
Compound of the present invention is best preparation method be dissolved in podophyllotoxin 1mmol in the 20ml methylene dichloride; N-(the 1-oxygen base-2 that adds 1 normal 1mmol; 2; 6; 6-tetramethyl--oxygen-carbonyl)-and the L-amino-acid compound, with N, the N-lutidine is as catalyzer under nitrogen protection; and the dicyclohexylcarbodiimide (DCC) that adds equivalent is dewatering agent, is reflected at room temperature condition and carries out.
Compound of the present invention can be used for preparing antitumor drug.
By compound of the present invention as seen, it is 4 on the C of Podophyllinic Acid Lactone ring to be replaced with N-(1-oxygen base-2,2,6,6-tetramethyl--oxygen-carbonyl)-L-amino acids form the Podophyllum emodi var chinense ester.Because Podophyllum emodi var chinense class medicine has multiple mechanism of action.The tubulin combination of podophyllotoxin energy and purifying suppresses the polymerization of cell microtubule, causes the karyomit(e) after duplicating not divide, and cell fission stops; And people such as Emanuel once reported 2,2,6,6-tetramethyl--4-hydroxy piperidine nitroxyl free radical has antileukemie effect, it is incorporated into plays synergy in the cancer therapy drug probably, many scholars will introduce nitroxyl free radical in the different cancer therapy drug molecules, and the toxicity of parent compound is reduced, and antitumour activity keeps even strengthen, and in our previous work nitroxyl free radical is introduced and has also found this point in the Podophyllum emodi var chinense molecule.Simultaneously, nitroxyl free radical can impel the delivery medicine preferentially to pass the cancerous tumor cell film as the carrier of pharmaceutical group, arrive cell DNA, and itself does not have toxicity and mutagen.On the other hand, L-amino acid can initiatively transmit in vivo, can be used as kinetophore is connected on the pharmacophore, and tumour is very fast in the speed of certain etap synthetic protein, require amino acid in cancer cells, to concentrate fast, help absorbing and transhipment, therefore, the present invention is with N-(1-oxygen base-2,2,6,6-tetramethyl--oxygen-carbonyl)-the amino acid whose compound of L-is incorporated into the expection of Podophyllum emodi var chinense molecule the toxicity of parent compound will be reduced by forming the mode that forms carboxylicesters with the C-4 position, and antitumour activity keeps even improve; And if use proper, also may reduce or delay chemical sproof generation, this compounds is also for to provide a kind of new tool by the anticancer mechanism of spectrum (ESR) the technical study about thing of Podophyllum emodi var chinense class simultaneously, Podophyllum emodi var chinense analog derivative and nitroxyl free radical advantage separately in addition, the former has significant cytotoxic activity and lipotropy thereof, and the latter can regulate characteristics such as drug molecule toxicity and good water-solubility thereof as pharmaceutical carrier; Add L-amino acid and in human body, have special physiologically active, therefore Podophyllum emodi var chinense derivative and nitroxyl free radical are coupled to a new drug molecule by L-amino acid, can obtain high-efficiency low-toxicity, lipid is suitable, the medicine that can fix a point to discharge particularly.Drug molecule is relatively stable in the transportation in vivo like this, decomposes gradually, discharges in tumour cell target spot location at last, reaches better result of treatment.And the present invention also may to overcome the first pass metabolism of podophyllum kind compound remarkable, lipotropy is low, oral absorption is incomplete, it is poorer that rectum absorbs, the deficiency that bioavailability is low, and overcome the shortcomings such as nausea,vomiting,diarrhea and alopecia of taking medicine to a certain extent.Correlation test of the present invention proves this compounds to the P-388 mouse leukemia, and A-549 people's lung cancer growth of tumour cell has significant inhibitory effect, and antitumor action belongs to potent.Compare with existing antineoplastic compound, compound of the present invention contains water soluble amino-acid and nitroxyl free radical part, does not but contain the complex construction of glucose.In addition, the synthetic method of compound of the present invention is simple, and raw material is cheap and easy to get.
According to relevant studies show that; the damage of DNA may cause sudden change and canceration in the organism; physics often can bring out the very active free radical of generation with factor chemistry; carcinogens also can generate free radical C through cell activation; these free radicals can make target molecule DNA responsive in the cell be transformed into DNA; form the DNA-C adducts with C subsequently; if these adductss may be the key factors that the stage is brought out in canceration. non-carcinogenic thing N and carcinogens are competed and are generated the DNA-N adducts, just may stop canceration.Existing report DNA can form adducts with nitroxyl free radical.And in the compound of the present invention owing to there is the N-O free radical, make the DNA-N adducts become possibility, thereby may stop canceration; Moreover, some discover that nitroxyl free radical can be as the carrier of pharmaceutical group, impel medicine preferentially to pass the cancerous tumor cell film, arrive cell DNA, it also has good water-solubility simultaneously and itself does not have toxicity and mutagen, add L-amino acid and have special advantages such as physiologically active in human body, these are indicating that all compound of the present invention will have higher anti-cancer activity, and this point also by after the correlation test chatted prove.
The result shows through the anti tumor activity in vitro screening study, the compound of formula I is to the P-388 mouse leukemia, A-549 people's lung cancer growth of tumour cell has significant inhibitory effect, and antitumor action belongs to potent, and therefore compound of the present invention can be used for preparing anti-tumor drug.
Embodiment
The preparation method of compound of the present invention and the activity experiment result of corresponding product below are provided
Preparation method of the present invention is: with podophyllotoxin and N-(1-oxygen base-2; 2; 6; 6-tetramethyl--oxygen-carbonyl)-L-amino acid is dissolved in the exsiccant methylene dichloride; the N that adds catalytic amount; the N-lutidine; under nitrogen protection, stirred 5 minutes; add dicyclohexylcarbodiimide (DCC) then, stirring reaction is 2 hours under nitrogen protection, removes by filter white precipitate; after removing solvent under reduced pressure; crude product through column chromatography purification with volume ratio is: 15: 1 methylene dichloride-acetone wash-out gets formula I compound.
Relevant reaction is referring to reaction formula 1.
Reaction formula 1
R in the formula 1 can be a hydrogen, or methyl, or sec.-propyl, or first sulphur methylene radical, or new butyl, or isobutyl-, or benzyl, or methylol, or the 2-hydroxyethyl, or to hydroxybenzyl.
Used N-(1-oxygen base-2,2,6,6-tetramethyl--oxygen-the carbonyl)-L-amino acid of the present invention obtains by following method:
With 2,2,6,6-tetramethyl--4-hydroxy piperidine is dissolved in 20ml water and 20ml methyl alcohol, add then 0.30 the gram sodium wolframate and 0.20 the gram disodium EDTA be placed on the magnetic stirring apparatus, begin to stir, add 30% hydrogen peroxide after waiting to dissolve in batches, continue to stir after 8-10 hour, get orange solid 2,2,6,6-tetramethyl--4-hydroxy piperidine nitroxyl free radical. will be dissolved in 2,2,6 of dry ether, 6-tetramethyl--4-hydroxy piperidine nitroxyl free radical dropwise add stirring be dissolved in dry tetrahydrofuran N, N '-carbonyl dimidazoles stirring at room 3 hours, forms N-(1-oxygen base-2,2,6,6-tetramethyl--oxygen-carbonyl)-imidazoles, the N-that is generated (1-oxygen base-2,2,6,6-tetramethyl--oxygen-carbonyl)-imidazoles joins in the exsiccant acetone soln that is dissolved with a water-toluene sulfonic acid, formed active sulfonate, this sulfonate is joined immediately in the aqueous solution that is dissolved with sodiumazide of stirring, at room temperature stirred 15 minutes, and formed 1-oxygen base-2,2,6,6-tetramethyl--oxygen-carbonyl azide is dissolved in this compound in the diox, it is joined be dissolved with in the amino acid and the magnesian aqueous solution, temperature is controlled under 40 ℃, stirred 24 hours, and used the ethyl acetate extraction drying then, steaming desolventizes purifying and obtains N-(1-oxygen base-2,2,6,6-tetramethyl--oxygen-carbonyl)-L-amino acid. method therefor is referring to literature method (H.A.Staab et al, Chem.Ber.1962,95,1284 reach H.0.HANKOVSZKY et al, Synthesis.1979,530-531); Its reaction is referring to following formula
Reaction formula 2
R in its reaction formula can be a hydrogen, or methyl, or sec.-propyl, or first sulphur methylene radical, or new butyl, or isobutyl-, or benzyl, or methylol, or the 2-hydroxyethyl, or to hydroxybenzyl.
The used podophyllotoxin of the present invention is to separate and be used as starting raw material from Chinese medicine Chinese podophyllum root (Podophyllium emodi Wallvar.Chinesis Spraque).Method therefor is referring to (Peter H.Hofert; Rudolf Matusch.Helvetica Chimica Acta.1994,77,771-777.)
Best preparation method of the present invention is dissolved in podophyllotoxin 1mmol in the 20ml methylene dichloride; N-(the 1-oxygen base-2 that adds 1 normal 1mmol; 2; 6; 6-tetramethyl--oxygen-carbonyl)-and the L-amino-acid compound, with N, the N-lutidine is as catalyzer under nitrogen protection; and the dicyclohexylcarbodiimide (DCC) that adds equivalent is dewatering agent, is reflected at room temperature condition and carries out.
The embodiment of the invention below is provided
Embodiment 1
4 α-4-L-glycine-N-(formyl-2 ', 2 ', 6 ', 6 '-tetramethyl--4 '-the nitroxyl free radical ester group) Podophyllum emodi var chinense ester (Ia) synthetic
In the Erlenmeyer flask of 250ml, add 20ml water and 20ml methyl alcohol, 0.30 gram sodium wolframate and 0.20 gram disodium EDTA, vibration dissolving, add 10 grams 2 then, 2,6,6-tetramethyl--4-hydroxy piperidine, be placed on the magnetic stirring apparatus, begin to stir, add 30% hydrogen peroxide after waiting to dissolve in batches, continue to stir, solution is by the colourless yellow that becomes, deepen gradually subsequently, become orange at last, react about 8-10 hour after, remove methyl alcohol and most of water under reduced pressure, debris cooling back adds a small amount of yellow soda ash vibration, and adds sodium-chlor and saltout, with extracted with diethyl ether 3-4 time, combining extraction liquid Calcium Chloride Powder Anhydrous drying, ether is removed in decompression, and cooling solid gets orange solid 2,2,6,6-tetramethyl--4-hydroxy piperidine nitroxyl free radical 10.5 grams.This reactions steps is that people such as Zhang Ziyi invent in Lanzhou University's journal (natural science edition) 1980,2,112.
To be dissolved in 1.72 grams 2 of 20ml dry ether, 2,6,6-tetramethyl--4-hydroxy piperidine nitroxyl free radical dropwise adds the 1.62 gram N that are dissolved in the 30ml dry tetrahydrofuran of stirring, N '-carbonyl dimidazoles, stirring at room 3 hours, water and sodium chloride saturated solution extraction respectively then, extraction liquid is by anhydrous sodium sulfate drying, evaporated under reduced pressure, recrystallization obtains N-(1-oxygen base-2,2,6 in ether/normal hexane, 6-tetramethyl--oxygen-carbonyl)-and imidazoles 1.6 grams. this reactions steps and H.A.Staab et al, Chem.Ber.1962,95,1284 disclosed methods are identical.
The N-that is generated (1-oxygen base-2,2,6,6-tetramethyl--oxygen-carbonyl)-and imidazoles 1.4 gram is dissolved in and joins in the exsiccant acetone in the exsiccant acetone soln that is dissolved with a water-toluene sulfonic acid 1.85 grams, stirs 10 minutes, and add ether then and occur precipitating, suction filtration has obtained active N-(1-oxygen base-2,2,6,6-tetramethyl--oxygen-carbonyl)-imidazoles sulfonate 1.2 grams.
These sulfonate 4.38 grams are joined in the 10ml aqueous solution that is dissolved with 1.3 gram sodiumazide of stirring immediately, at room temperature stirred 15 minutes, use n-hexane extraction then, and the extraction liquid anhydrous sodium sulfate drying, removal of solvent under reduced pressure is cooled to debris-20 ℃, get red crystals 1-oxygen base-2,2,6,6-tetramethyl--oxygen-carbonyl azide 2.17 grams.
With 2.41 gram 1-oxygen bases-2,2,6,6-tetramethyl--oxygen-carbonyl azide is dissolved in the 20ml diox, with its join be dissolved with 0.01mol amino acid and 0.8 the gram the magnesian 10ml aqueous solution in, temperature is controlled under 40 ℃, stirs 24 hours, also uses anhydrous sodium sulfate drying with ethyl acetate extraction then, steaming desolventizes purifying and obtains N-(1-oxygen base-2,2,6,6-tetramethyl--oxygen-carbonyl)-L-amino acid.This reactions steps is with reference to H.O.HANKOVSZKY et al, Synthesis.1979,530-531 institute reported method.
Podophyllotoxin is to separate and be used as starting raw material from Chinese medicine Chinese podophyllum root (Podophyllium emodi Wallvar.Chinesis Spraque).Method therefor is referring to (Peter H.Hofert; Rudolf Matusch.Helvetica Chimica Acta.1994,77,771-777.)
N-(1-oxygen base-2 with 1mmol podophyllotoxin and 1mmol; 2; 6; 6-tetramethyl--oxygen-carbonyl)-the sweet amino acid of L-is dissolved in the 20ml exsiccant methylene dichloride; add 0.1 gram N of catalytic amount, the N-lutidine stirred 5 minutes under nitrogen protection; add 0.21 gram dicyclohexylcarbodiimide (DCC) then; stirring reaction is 2 hours under nitrogen protection, removes by filter white precipitate, remove solvent under reduced pressure after; crude product is through column chromatography purification; with volume ratio is methylene dichloride-acetone wash-out of 15: 1,4 α-4-L-glycine-N-(formyl-2 ', 2 '; 6 ', 6 '-tetramethyl--4 '-the nitroxyl free radical ester group) the Podophyllum emodi var chinense ester.
The detection data of product are as follows:
Productive rate: 92%; M.p.135-137; [α]
D 25 ℃=-75 ° of (c=0.5, CH
2Cl
2) IR (KBr) υ cm
1: 3373,1780,1723,1485,1507,1589,930,1125,1179,1240,1371; MS (FAB) m/z:669 (M, 38), 397 (100); HRMS (ESI) C
34H
41N
2O
12: theoretical value (M+2H), 671.2811, measured value, 671.2802.ESR:g
0=2.0058, (3) Δ H
PP=35.309Gs, A
N(three peaks are 1 * 100 for=15.81Gs
-4M, CH
2Cl
2)
Embodiment 2
4 α-4-L-L-Ala-N-(formyl-2 ', 2 ', 6 ', 6 '-tetramethyl--4 '-the nitroxyl free radical ester group) Podophyllum emodi var chinense ester (Ib) synthetic
Experimental procedure and embodiment 1 only replace glycine with L-Ala together.It is as follows that the reaction products therefrom detects data:
Productive rate: 90%; M.p.138-140 ℃; [α]
D 25 ℃=-68 ° of (c=0.5, CH
2Cl
2) IR (KBr) υ cm
1: 3344,17811716,1485,1507,1589,930,1126,1175,1239,1365; MS (FAB) m/z:683 (M, 30), 397 (100); HRMS (ESl) C
35H
43N
2O
12Theoretical value (M+2H), 685.2967, measured value, 685.2962.ESR:g
0=2.0058, (3) Δ H
PP=44.268Gs, A
N(three peaks are 1 * 10 for=15.81Gs
4M, CH
2Cl
2)
Embodiment 3
4 α-4-L-methionine(Met)-N-(formyl-2 ', 2 ', 6 ', 6 '-tetramethyl--4 '-the nitroxyl free radical ester group) Podophyllum emodi var chinense ester (Ic) synthetic
Experimental procedure and embodiment 1 only replace the glycine of embodiment 1 together with methionine(Met).It is as follows that the reaction products therefrom detects data:
Productive rate: 91%; M.p.115-117; [α]
D 25 ℃=-65 ° of (c=0.5, CH
2Cl
2) IR (KBr) υ cm
1: 3337,1781,1719,1485,1507,1589,930,1126,1175,1239,1365; MS (FAB) m/z:743 (M
+, 68), 397 (100); HRMS (ESI) C
37H
47N
2O
12S: theoretical value (M+2H), 745.3001, measured value, 745.3001.ESR:g
0=2.0058, (3) Δ H
PP=34.255Gs, A
N(three peaks are 1 * 10 for=15.81Gs
-4M, CH
2Cl
2)
Embodiment 4
4 α-4-L-Isoleucine-N-(formyl-2 ', 2 ', 6 ', 6 '-tetramethyl--4 '-the nitroxyl free radical ester group) Podophyllum emodi var chinense ester (Id) synthetic
Experimental procedure and embodiment 1 only replace the glycine of embodiment 1 together with Isoleucine.It is as follows that the reaction products therefrom detects data:
Productive rate: 83%; M.p.117-118 ℃; [α]
D 25 ℃=-58 ° of (c=0.5, CH
2Cl
2) IR (KBr) υ cm
1: 3349,1782,1718,1485,1507,1589,930,1126,1178,1240,1366.MS (FAB) m/z:725 (M, 48), 397 (100); HRMS (ESI) C
38H
49N
2O
12: theoretical value (M+2H), 727.3437, measured value, 727.3437.ESR:g
0=2.0058, (3) Δ H
PP=32.674Gs, A
N(three peaks are 1 * 10 for=15.81Gs
-4M, CH
2C
L2)
Embodiment 5
4 α-4-L-phenylalanine-N-(formyl-2 ', 2 ', 6 ', 6 '-tetramethyl--4 '-the nitroxyl free radical ester group) Podophyllum emodi var chinense ester (Ie) synthetic
Experimental procedure and embodiment 1 only replace the glycine of embodiment 1 together with phenylalanine.It is as follows that the reaction products therefrom detects data:
Productive rate: 88%; M.p.120-121 ℃; [α]
D 25 ℃=-53 ° of (c=0.5, CH
2Cl
2) IR (KBr) υ cm
-1: 3334,1780,1719,1485,1506,1589,929,1125,1177,1239,1365.MS (FAB) m/z:759 (M, 27), 397 (100); HRMS (ESI) C
11H
17N
2O
12, theoretical value (M+2H), 761.3280, measured value, 761.3284.ESR:g
0=2.0058, (3) Δ H
PP=27.404Gs, A
N(three peaks are 1 * 10 for=15.81Gs
-4M, CH
2Cl
2)
Relevant pharmacological experimental method and result
Tetrazolium reduction method (MTT analytical method) is adopted in reason approximately of the present invention experiment, and (4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) is the basis with metabolism reduction 3-.Have the desaturase relevant with NADP in the plastosome of viable cell in experiment, xanchromatic MTT can be reduced to insoluble hepatic (Formazan), this enzyme of dead cell disappears, and is not reduced.Detect absorbancy at 550nm wavelength place with available microplate reader behind the DMSO dissolving Formazan.
Sulphonyl rhodamine B (sulforhodamineB, SRB) protein staining method SRB is a kind of protein binding dyestuff, can combine with the basic aminoacids in the biomacromolecule, it is good linear relationship at the OD of 515nm reading and cell count, so can be used as the quantitative of cell count.
Experimental technique is: (1) is got and was cultivated 4-5 days, is in one bottle of the cell cultures of exponential phase of growth, adds an amount of Trypsin-EDTA liquid, and attached cell is come off, and the RPMI1640 nutrient solution that contains 5% foetal calf serum with 10ml is made into suspension.(2) on blood cell counting plate, do the cell numeration with trypan blue dyeing back.(3) with cell culture medium diluting cells suspension, be made into every 100ml and contain 10000 or 20000 cells.(4) get 96 hole flat boards, every hole adds cell suspension 100ul.And flat board put 37 ℃ of CO
2(5%) incubator is 24 hours.(5) test-compound is made 5 extent of dilution, is followed successively by 10
1, 10
-5, 10
-6, 10
7, 10
8Mol/L.(6) with flat board at 37 ℃, contain 5%CO
2Hatched in the incubator of air and 100% humidity 2-3 days.(7) MTT is made into 1mg/ml solution with serum-free RPMI1640 nutrient solution, and every hole adds 50ul, and 37 incubations 4 hours make MTT be reduced to first .(8) draw supernatant liquor, add 150ulDMSO and make first dissolving, shake up with dull and stereotyped shaking table.(9) with the absorbancy of the dull and stereotyped reader of automatization spectrophotometric at 550nm place each aperture of mensuration.(10)
External The pharmacological results sees Table 1 The compounds of this invention to the influence to the inhibiting rate of A-549 people's lung cancer growth of tumour cell of the influence of the inhibiting rate of P-388 mouse leukemia growth of tumour cell and table 2 The compounds of this invention.
The inhibiting rate % of table 1 pair P-388 mouse leukemia growth of tumour cell
| The concentration sample number into spectrum | ????10 4 | ????10 5 | ????10 6 | ????10 7 | ????10 -8 | Estimate |
| ??Ia | ????96.3 | ????91.0 | ????89.1 | ????87.8 | ????77.5 | Potent |
| ??Ib | ????99.9 | ????89.2 | ????88.3 | ????81.1 | ????70.8 | Potent |
| ??Ic | ????94.9 | ????88.8 | ????86.3 | ????73.8 | ????43.0 | Potent |
| ??Id | ????95.8 | ????94.3 | ????73.9 | ????14.0 | ????0 | The weak effect |
| ??Ie | ????93.5 | ????92.6 | ????80.7 | ????68.6 | ????52.3 | Potent |
| ??VP-16 | ????99.2 | ????70.6 | ????25.5 | ????6.0 | ????1.8 | - |
Annotate: (1) screening method: tetrazolium reduction method; (2) action time: 72 hours; (3) sample number into spectrum Ia to Ie is respectively previous embodiment 1 to embodiment 5 products therefrom.
The inhibiting rate % of table 2 pair A-549 people's lung cancer growth of tumour cell
| The concentration sample number into spectrum | ????10 4 | ????10 5 | ????10 6 | ????10 7 | ????10 8 | Estimate |
| ??Ia | ????87.9 | ????87.3 | ????82.8 | ????80.1 | ????75.5 | Potent |
| ??Ib | ????94.7 | ????86.1 | ????83.5 | ????81.6 | ????50.6 | Potent |
| ??Ic | ????89.5 | ????83.5 | ????83.3 | ????73.2 | ????45.9 | Potent |
| ??Id | ????87.0 | ????83.6 | ????72.8 | ????39.0 | ????10.7 | The weak effect |
| ??Ie | ????87.9 | ????87.0 | ????81.9 | ????79.9 | ????60.0 | Potent |
| ??VP-16 | ????97.5 | ????50.7 | ????45.0 | ????0 | ????0 | - |
Annotate: (1) screening method: sulphonyl rhodamine B protein staining method; (2) action time: 72 hours; (3) sample number into spectrum Ia to Ie is respectively previous embodiment 1 to embodiment 5 products therefrom.
Experiment in vitro proves, in the compound wherein of the present invention four to leukemia cell line (P-388), people's lung cancer (A-549) has potent, its inhibiting rate is more much better than than VP-16, have only a compound to show as weak effect, its pharmacologically active is remarkable, and with the VP-16 contrast, compound of the present invention has more significant effect to leukemia cell line (P-388) and lung cancer cell line (A-549).The activity that can infer this compounds from this compounds is relevant with the amino acid whose type of L-probably, so be necessary to do in this respect further exploration.In addition from previous embodiment as seen, the synthetic method of such compound is simple, raw material is cheap and easy to get.
Claims (4)
1, as shown in the formula compound:
Wherein R can be a hydrogen, or methyl, or sec.-propyl, or first sulphur methylene radical, or new butyl, or isobutyl-, or benzyl, or methylol, or the 2-hydroxyethyl, or to hydroxybenzyl.
2, the preparation method of compound according to claim 1; it is characterized in that podophyllotoxin and N-(1-oxygen base-2; 2; 6; 6-tetramethyl--oxygen-carbonyl)-L-amino acid is dissolved in the exsiccant methylene dichloride; under nitrogen protection, stir; add dicyclohexylcarbodiimide (DCC) then; stirring reaction under nitrogen protection; reaction refilters after finishing and removes white precipitate, remove solvent under reduced pressure after, crude product is through column chromatography purification; with volume ratio is methylene dichloride-acetone wash-out of 15: 1, product.
3, method according to claim 2; it is characterized in that podophyllotoxin 1mmol is dissolved in the 20ml methylene dichloride; N-(the 1-oxygen base-2 that adds 1 normal 1mmol; 2,6,6-tetramethyl--oxygen-carbonyl)-the L-amino-acid compound; under nitrogen protection with N; the N-lutidine is as catalyzer, and the dicyclohexylcarbodiimide (DCC) that adds equivalent is dewatering agent, is reflected at room temperature condition and carries out.
4, the described compound of claim 1 is used to prepare antitumor drug.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2005100426486A CN100503612C (en) | 2005-04-28 | 2005-04-28 | Podophyllotoxin compounds and their application and preparation process |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104945409A (en) * | 2015-05-15 | 2015-09-30 | 东华大学 | Prodrug of compound podophyllotoxin PPT with anti-tumour activity and preparation method thereof |
| CN110294764A (en) * | 2019-07-15 | 2019-10-01 | 中国科学院兰州化学物理研究所 | A kind of podophyllotoxin derivative and preparation method thereof of azo key connection |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104945409A (en) * | 2015-05-15 | 2015-09-30 | 东华大学 | Prodrug of compound podophyllotoxin PPT with anti-tumour activity and preparation method thereof |
| CN110294764A (en) * | 2019-07-15 | 2019-10-01 | 中国科学院兰州化学物理研究所 | A kind of podophyllotoxin derivative and preparation method thereof of azo key connection |
| CN110294764B (en) * | 2019-07-15 | 2021-04-20 | 中国科学院兰州化学物理研究所 | Azo bond-connected podophyllotoxin derivative and preparation method thereof |
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