CN101891768A - 9-(diethoxy) phosphonomethyl purine, preparation method and application thereof in antineoplastic aspect - Google Patents
9-(diethoxy) phosphonomethyl purine, preparation method and application thereof in antineoplastic aspect Download PDFInfo
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- 238000002360 preparation method Methods 0.000 title claims abstract description 10
- FUXOPXSTMXZZMH-UHFFFAOYSA-N P(=O)(O)(O)CC1=NC=C2NC=NC2=N1 Chemical compound P(=O)(O)(O)CC1=NC=C2NC=NC2=N1 FUXOPXSTMXZZMH-UHFFFAOYSA-N 0.000 title abstract 3
- 230000000118 anti-neoplastic effect Effects 0.000 title abstract 2
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims abstract description 39
- -1 acyclic nucleoside phosphate compound Chemical class 0.000 claims abstract description 26
- 238000006243 chemical reaction Methods 0.000 claims abstract description 24
- 150000001875 compounds Chemical class 0.000 claims abstract description 20
- 239000002904 solvent Substances 0.000 claims abstract description 15
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims abstract description 14
- NZZFYRREKKOMAT-UHFFFAOYSA-N diiodomethane Chemical compound ICI NZZFYRREKKOMAT-UHFFFAOYSA-N 0.000 claims abstract description 11
- BDZBKCUKTQZUTL-UHFFFAOYSA-N triethyl phosphite Chemical compound CCOP(OCC)OCC BDZBKCUKTQZUTL-UHFFFAOYSA-N 0.000 claims abstract description 11
- BWHMMNNQKKPAPP-UHFFFAOYSA-L potassium carbonate Chemical compound [K+].[K+].[O-]C([O-])=O BWHMMNNQKKPAPP-UHFFFAOYSA-L 0.000 claims abstract description 10
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 7
- 239000003054 catalyst Substances 0.000 claims abstract description 7
- WQDUMFSSJAZKTM-UHFFFAOYSA-N Sodium methoxide Chemical compound [Na+].[O-]C WQDUMFSSJAZKTM-UHFFFAOYSA-N 0.000 claims abstract description 6
- 229910000027 potassium carbonate Inorganic materials 0.000 claims abstract description 5
- 239000011782 vitamin Substances 0.000 claims description 35
- 229940088594 vitamin Drugs 0.000 claims description 35
- 239000007788 liquid Substances 0.000 claims description 17
- LFGREXWGYUGZLY-UHFFFAOYSA-N phosphoryl Chemical group [P]=O LFGREXWGYUGZLY-UHFFFAOYSA-N 0.000 claims description 17
- 238000004821 distillation Methods 0.000 claims description 16
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 15
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 15
- UCQFCFPECQILOL-UHFFFAOYSA-N diethyl hydrogen phosphate Chemical compound CCOP(O)(=O)OCC UCQFCFPECQILOL-UHFFFAOYSA-N 0.000 claims description 15
- 229910052740 iodine Inorganic materials 0.000 claims description 15
- 239000011630 iodine Substances 0.000 claims description 15
- 229930003231 vitamin Natural products 0.000 claims description 13
- 235000013343 vitamin Nutrition 0.000 claims description 13
- 239000002253 acid Substances 0.000 claims description 12
- 239000000741 silica gel Substances 0.000 claims description 10
- 229910002027 silica gel Inorganic materials 0.000 claims description 10
- 230000008030 elimination Effects 0.000 claims description 7
- 238000003379 elimination reaction Methods 0.000 claims description 7
- 238000003756 stirring Methods 0.000 claims description 7
- 150000003722 vitamin derivatives Chemical class 0.000 claims description 7
- 230000001476 alcoholic effect Effects 0.000 claims description 6
- 239000012442 inert solvent Substances 0.000 claims description 6
- 238000004440 column chromatography Methods 0.000 claims description 5
- 239000007787 solid Substances 0.000 claims description 5
- 239000000725 suspension Substances 0.000 claims description 5
- 238000010189 synthetic method Methods 0.000 claims description 5
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 4
- 239000002246 antineoplastic agent Substances 0.000 claims description 3
- 229940041181 antineoplastic drug Drugs 0.000 claims description 3
- 230000006837 decompression Effects 0.000 claims description 2
- 239000000706 filtrate Substances 0.000 claims description 2
- 239000000463 material Substances 0.000 claims description 2
- 229910052757 nitrogen Inorganic materials 0.000 claims description 2
- 230000002829 reductive effect Effects 0.000 claims description 2
- 239000002002 slurry Substances 0.000 claims description 2
- 150000002500 ions Chemical class 0.000 claims 1
- 239000000126 substance Substances 0.000 claims 1
- 239000002777 nucleoside Substances 0.000 abstract description 10
- 229910019142 PO4 Inorganic materials 0.000 abstract description 9
- 239000010452 phosphate Substances 0.000 abstract description 9
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 abstract description 6
- 230000002401 inhibitory effect Effects 0.000 abstract description 6
- GFFGJBXGBJISGV-UHFFFAOYSA-N Adenine Chemical compound NC1=NC=NC2=C1N=CN2 GFFGJBXGBJISGV-UHFFFAOYSA-N 0.000 abstract 1
- 229930024421 Adenine Natural products 0.000 abstract 1
- 229960000643 adenine Drugs 0.000 abstract 1
- 206010073071 hepatocellular carcinoma Diseases 0.000 abstract 1
- 239000002994 raw material Substances 0.000 abstract 1
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- 201000007270 liver cancer Diseases 0.000 description 7
- 208000014018 liver neoplasm Diseases 0.000 description 7
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- 229940002612 prodrug Drugs 0.000 description 4
- 239000000651 prodrug Substances 0.000 description 4
- 230000000452 restraining effect Effects 0.000 description 4
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- HZVOZRGWRWCICA-UHFFFAOYSA-N methanediyl Chemical compound [CH2] HZVOZRGWRWCICA-UHFFFAOYSA-N 0.000 description 3
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- 239000008194 pharmaceutical composition Substances 0.000 description 3
- 239000000843 powder Substances 0.000 description 3
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- XYFCBTPGUUZFHI-UHFFFAOYSA-N Phosphine Chemical compound P XYFCBTPGUUZFHI-UHFFFAOYSA-N 0.000 description 2
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- 125000003835 nucleoside group Chemical group 0.000 description 2
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- HBOMLICNUCNMMY-UHFFFAOYSA-N 1-[4-azido-5-(hydroxymethyl)oxolan-2-yl]-5-methylpyrimidine-2,4-dione Chemical compound O=C1NC(=O)C(C)=CN1C1OC(CO)C(N=[N+]=[N-])C1 HBOMLICNUCNMMY-UHFFFAOYSA-N 0.000 description 1
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 1
- 238000005160 1H NMR spectroscopy Methods 0.000 description 1
- 238000004679 31P NMR spectroscopy Methods 0.000 description 1
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- HAWPXGHAZFHHAD-UHFFFAOYSA-N mechlorethamine Chemical compound ClCCN(C)CCCl HAWPXGHAZFHHAD-UHFFFAOYSA-N 0.000 description 1
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- 229940087004 mustargen Drugs 0.000 description 1
- 238000006053 organic reaction Methods 0.000 description 1
- 150000003014 phosphoric acid esters Chemical class 0.000 description 1
- 229910000073 phosphorus hydride Inorganic materials 0.000 description 1
- 238000006366 phosphorylation reaction Methods 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 238000009790 rate-determining step (RDS) Methods 0.000 description 1
- 239000003419 rna directed dna polymerase inhibitor Substances 0.000 description 1
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- HBOMLICNUCNMMY-XLPZGREQSA-N zidovudine Chemical compound O=C1NC(=O)C(C)=CN1[C@@H]1O[C@H](CO)[C@@H](N=[N+]=[N-])C1 HBOMLICNUCNMMY-XLPZGREQSA-N 0.000 description 1
- 229960002555 zidovudine Drugs 0.000 description 1
Landscapes
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses an acyclic nucleoside phosphate compound, i.e. 9-(diethoxy) phosphonomethyl purine, a preparation method and application thereof in the antineoplastic aspect. The structural formula of the compound is shown as the following specification. In the preparation method, the 9-(diethoxy) phosphonomethyl purine is prepared by two reactions by using methylene diiodide, triethyl-phosphite and adenine as raw materials, using N,N-dimethyl formamide, dimethyl sulfoxide, methanol and ethanol as solvents and using anhydrous potassium carbonate and sodium methoxide as catalysts. The compound has a certain inhibiting effect on the hepatoma cell BEL-7402.
Description
Technical field:
The present invention relates to a kind of 9-(diethoxy) phosphonomethyl adenine compound; The invention still further relates to the synthetic method of described compound and in the application of anti-tumor aspect.
Background technology:
Tumour is a kind of disease of serious threat human health, do not find the radical cure anti-tumor drug so far as yet, but after nineteen forty-three, mustargen was used for the treatment of malignant lymphoma, anticancer chemotherapy has been obtained sizable progress in decades.In recent years, nucleoside medicine has been subjected to extensive concern as the main curative of virus type disease.Being used for the treatment of the nucleoside medicine that human immunodeficiency virus (HIV) and hsv (HSV) catch generally all is the inhibitor of HIV viral reverse transcriptase inhibitor and HSV vDNA (DNA) polysaccharase.Therefore, nucleoside medicine has good therapeutic action to many virus diseases.Though nucleosides and analogue thereof are widely used as antitumor and antiviral clinically, many medicines have presented toxic side effect in various degree, and its application is restricted.Discover and in drug molecule, introduce heteroatom group, be expected to reach the purpose that improves curative effect by producing synergy.Phosphide is the effective carrier of medicine, tumor tissues had certain targeting, people attempt nucleoside medicine is carried out structural modification, introducing phosphoric acid ester, will to show that it is converted into phosphatide be the prodrug of carrier, avoid the first step phosphorylation reaction of rate-limiting step, simultaneously the sugar ring is opened, keep compound conformation flexibility, developed a series of open loop nucleosides phosphine things thus, this has become the focus of antiviral research in recent years.
At present some have been carried out about the research of drug compound before the nucleoside phosphoric acid ester, also developed HIV has been had this type of prodrug [Bis (POM)-(PMEA)] and [Bis (POM)-(PMPA)] that efficient inhibition is done, but on the whole, the research of this respect also is in the starting stage, need more research to seek the nucleoside phosphoric acid ester prodrug of superior activity, the content of at present relevant nucleoside phosphoric acid ester prodrug 9-(diethoxy) phosphonomethyl VITAMIN B4 is not seen bibliographical information.Given this, the present invention reports to this compound and preparation method thereof with in the application of anti-tumor aspect.
Summary of the invention:
At the deficiencies in the prior art and this area research and demands of applications, the purpose of this invention is to provide a kind of acyclic nucleotide phosphate compound 9-(diethoxy) phosphonomethyl VITAMIN B4; The present invention simultaneously also provides the synthetic method of described all compounds and in the application of anti-tumor aspect.
The present invention is subjected to the inspiration of lead compound [Bis (POM)-(PMEA)] and [Bis (POM)-(PMPA)] structure, has synthesized acyclic nucleotide phosphate compound 9-(diethoxy) phosphonomethyl VITAMIN B4 by the two-step reaction design; And examined or check this compound to the tumour cell restraining effect.
(1) a kind of acyclic nucleotide phosphate compound 9-provided by the invention (diethoxy) phosphonomethyl VITAMIN B4, its structure is as follows:
(2) acyclic nucleotide phosphate compound 9-of the present invention (diethoxy) phosphonomethyl VITAMIN B4 is synthetic by two step organic reactions, and the main points of synthetic method are as follows:
The first step: alpha-iodine methyl acid phosphate diethyl ester synthetic
The methylene iodide adding is equipped with in the there-necked flask of prolong, behind heat temperature raising to 80～110 ℃, beginning drips triethyl-phosphite with the speed of 1～5 of per second in reaction flask, when being raised to 140 ℃, temperature removes thermal source rapidly, strict controlled temperature, make it not surpass 150 ℃, after 3～6 times of triethyl-phosphites to the methylene iodide volume are dropwised, termination reaction when temperature drops to 40～80 ℃; The faint yellow reaction solution of gained distills under the 13mmHg reduced pressure, when temperature has two kinds of cuts to be steamed 60-90 ℃ of priority, adjust vacuum tightness then, underpressure distillation under 8mmHg, when temperature rises to 106 ℃, the third cut is steamed, and remaining almost colourless liquid is alpha-iodine methyl acid phosphate diethyl ester;
Second step: 9-(diethoxy) phosphoryl methyl-VITAMIN B4 synthetic
VITAMIN B4 and the adding of anhydrous basic catalyst are equipped with in the there-necked flask of anhydrous inert solvent, the suspension liquid that forms under nitrogen protection in 70～150 ℃ stir 10～50 minutes after, with syringe 1～3 times of alpha-iodine methyl acid phosphate diethyl ester to the VITAMIN B4 molar weight is added drop-wise in the reaction solution, stirring reaction is 12～48 hours under 10～50 ℃ of conditions, the elimination insolubles, the solvent in the filtrate is removed in decompression, get yellow thickness slurry like material, add the alcoholic solvent dissolving, by diatomite elimination insolubles, add proper silica gel, underpressure distillation, make it be adsorbed in silica gel fully,, get 9-(diethoxy) phosphoryl methyl-VITAMIN B4 white waxy solid with ethyl acetate/methanol=8: 1 rapid column chromatographies;
Wherein, the anhydrous basic catalyst in the reaction of second step is Anhydrous potassium carbonate or sodium methylate; Anhydrous inert solvent is N, dinethylformamide or dimethyl sulfoxide (DMSO); Alcoholic solvent is methyl alcohol or ethanol.
In the preparation method of above-mentioned 9-(diethoxy) phosphoryl methyl-VITAMIN B4, the anhydrous basic catalyst in second step reaction in the reaction of second step is an Anhydrous potassium carbonate; Anhydrous inert solvent is N, dinethylformamide; Alcoholic solvent is a methyl alcohol.
In the building-up process of above-mentioned acyclic nucleotide phosphate compound 9-(diethoxy) phosphoryl methyl-VITAMIN B4, the reaction equation that relates to is as follows:
(3) structural characterization of 9-(diethoxy) phosphoryl methyl-VITAMIN B4:
Nuclear magnetic data:
1H-NMR (500MHz, DMSO-d
6): δ 1.14-1.17 (6H, t, J=7.5Hz, 2CH
3), 4.02-4.06 (4H, m, 2CH
2), 4.69-4.72 (2H, d, J=15.0Hz, POCH
2), 7.29 (2H, s, NH
2), 8.04 (1H, s, H-8), 8.17 (1H, s, H-2).
13C-NMR (500MHz, DMSO-d
6): δ 16.53 (2CH
3), 34.78-38.69 (d, J=605Hz, POCH
2), 62.92 (2CH
2), 118.46 (C-5), 141.06 (C-8), 149.86 (C-4), 153.09 (C-2), 156.42 (C-6).
31P-NMR (500MHz, DMSO-d
6): δ 18.95.
Ultimate analysis data: Anal.Calcd for C
12H
16N
5O
3P:C, 46.60; H, 5.21; N, 22.65, P, 10.02.Found:C, 46.72; H, 5.17; N, 22.74, P, 9.98.
Mass-spectrometric data: TOF-MS (ES+1.76e) m/z 286.1[M+H]
+.
(4) the present invention also provides the application of described acyclic nucleotide phosphate compound 9-(diethoxy) phosphoryl methyl-VITAMIN B4 in the preparation antitumor drug.
The present invention also provides and has been used for antitumor medicine composition, wherein contains one or more carriers of acyclic nucleotide phosphate compound 9-of the present invention (diethoxy) phosphoryl methyl-VITAMIN B4 for the treatment of significant quantity.
Suppressing tumour with acyclic nucleotide phosphate compound 9-of the present invention (diethoxy) phosphoryl methyl-VITAMIN B4 below, especially the inhibition experiment of human liver cancer cell BEL-7402 is further being set forth.
Adopt mtt assay, measure The compounds of this invention in external restraining effect to human liver cancer cell BEL-7402.In the anti-tumor activity research, be target cell with human liver cancer cell BEL-7402, cell culture medium is 1640 substratum that contain 10%FCS, the logarithmic phase cell with trysinization after, regulating cell density is 3～5 * 10
3Individual/hole, be seeded in 96 well culture plates, every hole 100 μ l, putting into temperature is 37 ℃, CO
2Content is to cultivate 24 hours in 0.5% the incubator.The sample of the different concns for preparing is added 100 μ l/ holes in 96 well culture plates, (put into temperature is 37 ℃ to the positive control drug zidovudine of different weaker concns, CO for Zidovudine, AZT) the positive contrast of soup
2Content is to continue in 0.5% the incubator to cultivate 48 hours.Take out 96 well culture plates, every hole adds the MTT of the 5mg/mL of 20 μ l, continues to cultivate 4 hours.Take out culture plate, pour out substratum, every hole adds 150 μ l DMSO, and 37 ℃ vibrated 6 minutes, and the 570nm place measures the optical density value in each hole.Calculate inhibiting rate: (OD
Negative control-OD
Sample)/OD
Negative control* 100%.
Compound of the present invention and pharmaceutical composition can be used for preparing the medicine of tumour.
The various formulations of pharmaceutical composition of the present invention can for example make activeconstituents mix with one or more carriers according to the conventional production method preparation of pharmaceutical field, are made into required formulation then.
Pharmaceutical composition of the present invention preferably contains the activeconstituents that mass ratio is 0.1%-99.5%.
According to synthetic route of the present invention and method, can stablize, repeatable the The compounds of this invention for preparing; Adopt method of the present invention to prepare described compound, easy and simple to handle, practical; The anti tumor activity in vitro evidence, The compounds of this invention has certain restraining effect to human liver cancer cell BEL-7402.
Embodiment:
Following examples can make those skilled in the art more fully understand the present invention, but do not limit the present invention in any way.Specify the preparation method of 9-of the present invention (diethoxy) phosphoryl methyl-VITAMIN B4 below by experiment, and this compound is in the application of anti-tumor method.
Embodiment 1:
(1) alpha-iodine methyl acid phosphate diethyl ester is synthetic
(81mL 1.00mol), is heated to 110 ℃ to add methylene iodide in the 500mL there-necked flask of condensation side pipe is housed, the color of methylene iodide begins to deepen, and drip triethyl-phosphite this moment slowly, and temperature rises rapidly in the dropping process, the reaction beginning is described, remove thermal source when temperature is raised to 140 ℃ rapidly, strict controlled temperature makes them can not be above 150 ℃, drip triethyl-phosphite (180mL, 1.05mol), begin to descend until temperature of reaction, illustrate to react and finish.After the cooling, get weak yellow liquid fully.With its underpressure distillation under 13mmHg, when temperature has two kinds of cuts to be steamed 70-90 ℃ of priority, adjust vacuum tightness then, underpressure distillation under 8mmHg, temperature rises to 106 ℃, the third cut is steamed, the product alpha-iodine methyl acid phosphate diethyl ester 239.08g that then remaining almost colourless liquid is gained, and productive rate is 86%.
(2) 9-(diethoxy) phosphoryl methyl-VITAMIN B4 is synthetic
In the there-necked flask of 50mL, add VITAMIN B4 (2.70g; 0.02mol); being suspended in 12mL heavily steams among the DMF; form the oyster white suspension liquid; the powder NaH of adding fresh dried (6.1g, 0.15mol), 80 ℃ of Ar gas shileds were stirred 30 minutes down; with syringe slowly drip alpha-iodine methylene radical diethyl phosphoric acid (5.56g, 0.02mol).Stirring at room 24 hours.The elimination insolubles removes neat solvent with clear liquid underpressure distillation under high vacuum, yellow utmost point thickness soup compound, add dissolve with methanol, by the siliceous earth column filter, go out insolubles.Add proper silica gel again, underpressure distillation makes it be adsorbed in silica gel fully.Sample on the solid adopts careful second component of collecting of ethyl acetate/methanol (8: 1) elutriant column chromatography, is spin-dried for solvent fast and gets white waxy solid product 9-(diethoxy) phosphoryl methyl-VITAMIN B4 3.19g, and productive rate is 56%.
Embodiment 2:
(1) alpha-iodine methyl acid phosphate diethyl ester is synthetic
(81mL 1.00mol), is heated to 110 ℃ to add methylene iodide in the 500mL there-necked flask of condensation side pipe is housed, the color of methylene iodide begins to deepen, and drip triethyl-phosphite this moment slowly, and temperature rises rapidly in the dropping process, the reaction beginning is described, remove thermal source when temperature is raised to 120 ℃ rapidly, strict controlled temperature makes them can not be above 130 ℃, drip triethyl-phosphite (180mL, 1.05mol), begin to descend until temperature of reaction, illustrate to react and finish.After the cooling, get weak yellow liquid fully.With its underpressure distillation under 13mmHg, when temperature has two kinds of cuts to be steamed 70-90 ℃ of priority, adjust vacuum tightness then, underpressure distillation under 8mmHg, temperature rises to 106 ℃, the third cut is steamed, the product alpha-iodine methyl acid phosphate diethyl ester 216.86g that then remaining almost colourless liquid is gained, and productive rate is 78%.
(2) 9-(diethoxy) phosphoryl methyl-VITAMIN B4 is synthetic
In the there-necked flask of 50mL, add VITAMIN B4 (2.70g; 0.02mol); being suspended in 12mL heavily steams among the DMF; form the oyster white suspension liquid; the powder NaH of adding fresh dried (6.1g, 0.15mol), 60 ℃ of Ar gas shileds were stirred 30 minutes down; with syringe slowly drip alpha-iodine methylene radical diethyl phosphoric acid (5.56g, 0.02mol).Stirring at room 24 hours.The elimination insolubles removes neat solvent with clear liquid underpressure distillation under high vacuum, yellow utmost point thickness soup compound, add dissolve with methanol, by the siliceous earth column filter, go out insolubles.Add proper silica gel again, underpressure distillation makes it be adsorbed in silica gel fully.Sample on the solid adopts careful second component of collecting of ethyl acetate/methanol (8: 1) elutriant column chromatography, is spin-dried for solvent fast and gets white waxy solid product 9-(diethoxy) phosphoryl methyl-VITAMIN B4 3.15g, and productive rate is 51%.
Embodiment 3:
(1) alpha-iodine methyl acid phosphate diethyl ester is synthetic
(81mL 1.00mol), is heated to 100 ℃ to add methylene iodide in the 500mL there-necked flask of condensation side pipe is housed, the color of methylene iodide begins to deepen, and drip triethyl-phosphite this moment slowly, and temperature rises rapidly in the dropping process, the reaction beginning is described, remove thermal source when temperature is raised to 120 ℃ rapidly, strict controlled temperature makes them can not be above 130 ℃, drip triethyl-phosphite (180mL, 1.05mol), begin to descend until temperature of reaction, illustrate to react and finish.After the cooling, get weak yellow liquid fully.With its underpressure distillation under 13mmHg, when temperature has two kinds of cuts to be steamed 70-90 ℃ of priority, adjust vacuum tightness then, underpressure distillation under 8mmHg, temperature rises to 106 ℃, the third cut is steamed, the product alpha-iodine methyl acid phosphate diethyl ester 197.40g that then remaining almost colourless liquid is gained, and productive rate is 71%.
(2) 9-(diethoxy) phosphoryl methyl-VITAMIN B4 is synthetic
In the there-necked flask of 50mL, add VITAMIN B4 (2.70g; 0.02mol); being suspended in 12mL heavily steams among the DMF; form the oyster white suspension liquid; the powder NaH of adding fresh dried (6.1g, 0.15mol), 80 ℃ of Ar gas shileds were stirred 30 minutes down; with syringe slowly drip alpha-iodine methylene radical diethyl phosphoric acid (5.56g, 0.02mol).Stirring at room 12 hours.The elimination insolubles removes neat solvent with clear liquid underpressure distillation under high vacuum, yellow utmost point thickness soup compound, add dissolve with methanol, by the siliceous earth column filter, go out insolubles.Add proper silica gel again, underpressure distillation makes it be adsorbed in silica gel fully.Sample on the solid adopts careful second component of collecting of ethyl acetate/methanol (8: 1) elutriant column chromatography, is spin-dried for solvent fast and gets white waxy solid product 9-(diethoxy) phosphoryl methyl-VITAMIN B4 2.97g, and productive rate is 48%.
Embodiment 4:
With human liver cancer cell BEL-7402 is target cell, and cell culture medium is 1640 substratum that contain 10%FCS, the logarithmic phase cell with trysinization after, regulating cell density is 3～5 * 10
3Individual/hole, be seeded in 96 well culture plates, every hole 100 μ l, putting into temperature is 37 ℃, CO
2Content is to cultivate 24 hours in 0.5% the incubator.The sample that the concentration for preparing is respectively 0.4mg/mL, 0.8mg/mL, 1.2mg/mL, 1.6mg/mL and 2.0mg/mL adds 100 μ l/ holes in 96 well culture plates, putting into temperature is 37 ℃, CO
2Content is to continue respectively in 0.5% the incubator to cultivate 12,24,36,48 hours.Take out 96 well culture plates, every hole adds the MTT of the 5mg/mL of 20 μ l, continues to cultivate 4 hours.Take out culture plate, pour out substratum, every hole adds 150 μ l DMSO, and 37 ℃ vibrated 6 minutes, and the 570nm place measures the optical density value in each hole.Calculate inhibiting rate: (OD
Negative control-OD
Sample)/OD
Negative control* 100%.
Table 1:9-(diethoxy) phosphoryl methyl-VITAMIN B4 is to the inhibiting rate of liver cancer cell BEL-7402
The result shows: compound 9-(diethoxy) phosphoryl methyl-VITAMIN B4 that the present invention tested has the obvious suppression effect to liver cancer cell BEL-7402.Its restraining effect presents certain time-concentration relationship, and promptly along with administration concentration is increased to 2.0mg/mL from 0.4mg/mL, its inhibiting rate to tumour cell has tangible increase; And along with the tumor cell culture time after the administration extended to 48 hours from 12 hours, with the concentration medicine inhibiting rate of tumour cell also there is tangible increase, shows that this compound has the relevant dependence of tangible time-concentration-drug effect aspect cytotoxicity.
Claims (4)
1. a 9-(diethoxy) phosphoryl methyl-adenine compound is characterized in that this ion liquid chemical structural formula is as follows:
2. the synthetic method of 9-described in the claim 1 (diethoxy) phosphoryl methyl-adenine compound, its feature target compound is synthetic through following steps:
The first step: alpha-iodine methyl acid phosphate diethyl ester synthetic
The methylene iodide adding is equipped with in the there-necked flask of prolong, behind heat temperature raising to 80～110 ℃, beginning drips triethyl-phosphite with the speed of 1～5 of per second in reaction flask, when being raised to 140 ℃, temperature removes thermal source rapidly, strict controlled temperature, make it not surpass 150 ℃, after 3～6 times of triethyl-phosphites to the methylene iodide volume are dropwised, termination reaction when temperature drops to 40～80 ℃; The faint yellow reaction solution of gained distills under the 13mmHg reduced pressure, when temperature has two kinds of cuts to be steamed 60-90 ℃ of priority, adjust vacuum tightness then, underpressure distillation under 8mmHg, when temperature rises to 106 ℃, the third cut is steamed, and remaining almost colourless liquid is alpha-iodine methyl acid phosphate diethyl ester;
Second step: 9-(diethoxy) phosphoryl methyl-VITAMIN B4 synthetic
VITAMIN B4 and the adding of anhydrous basic catalyst are equipped with in the there-necked flask of anhydrous inert solvent, the suspension liquid that forms under nitrogen protection in 70～150 ℃ stir 10～50 minutes after, with syringe 1～3 times of alpha-iodine methyl acid phosphate diethyl ester to the VITAMIN B4 molar weight is added drop-wise in the reaction solution, stirring reaction is 12～48 hours under 10～50 ℃ of conditions, the elimination insolubles, the solvent in the filtrate is removed in decompression, get yellow thickness slurry like material, add the alcoholic solvent dissolving, by diatomite elimination insolubles, add proper silica gel, underpressure distillation, make it be adsorbed in silica gel fully,, get 9-(diethoxy) phosphoryl methyl-VITAMIN B4 white waxy solid with ethyl acetate/methanol=8: 1 rapid column chromatographies;
In the synthetic method of above-mentioned 9-(diethoxy) phosphoryl methyl-VITAMIN B4, the anhydrous basic catalyst in the reaction of second step is Anhydrous potassium carbonate or sodium methylate; Anhydrous inert solvent is N, dinethylformamide or dimethyl sulfoxide (DMSO); Alcoholic solvent is methyl alcohol or ethanol.
3. the preparation method of 9-described in the claim 2 (diethoxy) phosphoryl methyl-VITAMIN B4 is characterized in that, the anhydrous basic catalyst in second step reaction in the reaction of second step is an Anhydrous potassium carbonate; Anhydrous inert solvent is N, dinethylformamide; Alcoholic solvent is a methyl alcohol.
4. 9-described in the claim 1 (diethoxy) phosphoryl methyl-VITAMIN B4 application of being used to prepare antitumor drug.
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| CN106831872A (en) * | 2017-01-10 | 2017-06-13 | 黄石福尔泰医药科技有限公司 | A kind of impurity of tenofovir or tenofovir disoproxil and preparation method thereof |
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| CN1938307A (en) * | 2004-03-26 | 2007-03-28 | 大日本住友制药株式会社 | 9-substituted 8-oxoadenine compound |
| CN101723827A (en) * | 2009-12-15 | 2010-06-09 | 上虞新和成生物化工有限公司 | Preparation method of 4-acetoxyl-2-methyl-2-butylenoic aldehyde |
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| CN1938307A (en) * | 2004-03-26 | 2007-03-28 | 大日本住友制药株式会社 | 9-substituted 8-oxoadenine compound |
| CN101723827A (en) * | 2009-12-15 | 2010-06-09 | 上虞新和成生物化工有限公司 | Preparation method of 4-acetoxyl-2-methyl-2-butylenoic aldehyde |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106831872A (en) * | 2017-01-10 | 2017-06-13 | 黄石福尔泰医药科技有限公司 | A kind of impurity of tenofovir or tenofovir disoproxil and preparation method thereof |
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