CN104622887A

CN104622887A - Antitumor drug prepared through combination of purine deoxynucleoside and other nucleosides or basic groups as well as preparation method and applications thereof

Info

Publication number: CN104622887A
Application number: CN201410675196.4A
Authority: CN
Inventors: 张始状; 韩明; 高志琴; 程鑫; 赵荣荣
Original assignee: Individual
Current assignee: Individual
Priority date: 2014-11-24
Filing date: 2014-11-24
Publication date: 2015-05-20
Also published as: CN106074590A

Abstract

The invention discloses an antitumor drug prepared through the combination of purine deoxynucleoside and other nucleosides or basic groups as well as a preparation method and applications thereof. The purine deoxynucleoside comprises deoxyadenosine and deoxyguanosine; and the other nucleosides or basic groups include deoxycytidine, thymidine, adenosine, guanosine, cytidine, uridine, adenine, guanine, cytosine, uracil, and thymine. The antitumor drug disclosed by the invention has the advantages that the purine deoxynucleoside and the other nucleosides or basic groups are all normal nucleosides of human bodies, the combination of purine deoxynucleoside as a main drug and other nucleosides or basic groups as an adjuvant is used for preparing a drug for treating various common malignant tumors such as gastric carcinoma, lung carcinoma, hepatic carcinoma, colorectal carcinoma, esophageal carcinoma, pancreatic carcinoma, breast carcinoma, genital system tumor, and the like, and the antitumor drug disclosed by the invention has the characteristics of broad-spectrum antitumor effect, low toxicity and new mechanism and the like; toxic and side effects caused by combined medication are reduced in comparison with single medication; the multiplicity of the antitumor effect is increased; and the occurrence of tumor drug-resistance can be delayed.

Description

Antitumor drug that deoxy purine nucleoside is prepared with other nucleoside or base composition and its preparation method and application

Technical field

The present invention relates to human normal deoxyadenosine and nucleoside or base composition and prepare the application in tumour medicine, specifically one or more in one or both in deoxyadenosine, deoxyguanosine and nucleoside or base are combined into medicine and are used for the treatment of tumor, the most classical is that two kinds of deoxy purine nucleoside prepare tumour medicine with other nucleoside a kind of or base composition.Belong to tumor new drug development field.

Background technology

Nucleoside (nucleoside) is formed by connecting by base and pentose (ribose or deoxyribose), the i.e. compound that is formed by connecting by β glycosidic bond of the C-1 of the N-9 of purine or the N-1 of pyrimidine and ribose or deoxyribose, comprise ribonucleotide and dezyribonucleoside two class, ribonucleotide forms RNA, mainly contains adenosine, guanosine, cytidine and uridnine; Dezyribonucleoside (abbreviation deoxynucleoside) forms DNA, mainly contains deoxyadenosine, deoxyguanosine, deoxycytidine and thymidine.Base is the pith of composition nucleoside, mainly contains adenine, guanine, cytosine, uracil and thymus pyrimidine.

Ribonucleotide, the compound be combined into by the purine except thymus pyrimidine or pyrimidine and ribose molecule covalent, mainly contains adenosine, guanosine, cytidine and uridnine.Ribosidoadenine (adenosine) is called for short adenosine, chemical name: 9-β-D-RIBOSE base adenine, No. CAS: 58-61-7, molecular formula: C ₁₀h ₁₃n ₅o ₄, molecular weight: 267.24.Guanosine (Guanosine) guanosine, chemical name: 9-β-D-ribofuranoside bird is fast, No. CAS: 118-00-3, molecular formula: C ₁₀h ₁₃n ₅o ₅, molecular weight: 283.24.Cytidine (Cytidine) is called for short cytidine, chemical name: 1-β-D-RIBOSE base cytosine, No. CAS: 65-46-3, molecular formula: C ₉h ₁₃n ₃o ₅, molecular weight 243.22.Uridine is called for short uridnine, chemical name: 1-β-D-RIBOSE base uracil, No. CAS: 58-96-8, molecular formula: C ₉h ₁₂n ₂o ₆, molecular weight 244.20.

Dezyribonucleoside (deoxynucleoside), be the compound that the N-9 of purine base (adenine, guanine) or the N-1 of pyrimidine base (cytosine, thymus pyrimidine) are connected by β glycosidic bond with the C-1 of DRI, deoxynucleoside main in body has deoxyadenosine, deoxyguanosine, deoxycytidine, thymidine (abbreviation thymidine).Deoxyadenosine (deoxyadenosine) i.e. deoxyadenosine, chemical name: 9-β-D-2 '-deoxidation ribofuranoside adenine, being the N-9 of adenine to be connected formed compound by β glycosidic bond with the C-1 of DRI, and No. CAS: 958-09-8, molecular formula: C ₁₀h ₁₃n ₅o ₃, molecular weight: 251.24.Deoxyguanosine (deoxyguanosine) i.e. deoxyguanosine, chemical name: 9-β-2 '-furan deoxynucleoside guanine, being the N-9 of guanine to be connected formed compound by β glycosidic bond with the C-1 of DRI, and No. CAS: 961-07-9, molecular formula: C ₁₀h ₁₃n ₅o ₄, molecular weight: 267.24.Deoxycytidine (deoxycytidine) i.e. deoxycytidine, chemical name: 1-β-2 '-furan deoxynucleoside cytosine, being the N-1 of cytosine to be connected formed compound by β glycosidic bond with the C-1 of DRI, and No. CAS: 951-77-9, molecular formula: C ₉h ₁₃n ₃o ₄, molecular weight: 227.22.Thymidine is called for short thymidine (Thymidine) i.e. Thymidine, or beta-thymidine, chemical name: 9-β-D-ribofuranoside thymus pyrimidine, No. CAS: 50-89-5, molecular formula: C ₁₀h ₁₄n ₂o ₅, molecular weight is 242.023.

Base is the part that nucleoside or deoxynucleoside remove ribose, comprising: adenine, guanine, cytosine, uracil and thymus pyrimidine.Adenine (adenine): also known as adenine phosphate, No. CAS: 73-24-5, molecular formula: C ₅h ₅n ₅, molecular weight: 135.127; Guanine (guanine): No. CAS: 73-40-5, molecular formula: C ₅h ₅n ₅o, molecular weight: 151.13; Cytosine (Cytosine): No. CAS: 71-30-7, molecular formula: C ₄h ₅n ₃o, molecular weight 111.10; Uracil (Uracil): No. CAS: 66-22-8, molecular formula: C ₄h ₄n ₂o ₂, molecular weight 112.09; Thymus pyrimidine (Thymine): No. CAS: 65-71-4, molecular formula: C ₅h ₆n ₂o ₂, molecular weight: 126.1133.

Deoxy-ribonucleoside monophosphate is the structure fragment forming DNA (deoxyribonucleic acid) DNA, and deoxynucleoside and derivant thereof have good physiologically active, are the important source material of genomic medicine and genetic engineering research.Ucleosides antitumor drug is a common class antineoplastic agent, only has purines 31 kinds in antimetabolitas, wherein goes on the market 10 kinds, and miazines 35 kinds wherein goes on the market 9 kinds.These medicines are parent with participating in the normal nucleotide class material of cellular metabolism, replaces normal element and group carries out structure of modification and modification with a kind of element or group, thus reach the object with anti-tumor activity.Such as: hypoxanthine, 5-fluorouracil etc.

The present invention and aforementioned antineoplastic agent essential distinction are: the various nucleoside related to and base are not abnormal substituent human normal nucleoside and base.Current people it is generally acknowledged that normal nucleotide can promote tumor growth, instead of Tumor suppression growth.Therefore, normal deoxy purine nucleoside and other nucleoside or base composition are prepared antitumor drug, there is not yet relevant report both at home and abroad.

Summary of the invention

The invention provides a kind of deoxy purine nucleoside and other nucleoside, base composition prepare tumour medicine, using a kind of or two kinds of deoxy purine glycosides with comprise adenosine, guanosine, cytidine, uridnine, deoxycytidine, thymidine, adenine, guanine, cytosine, uracil, thymus pyrimidine a kind of, two or more ingredients combined as treating tumor use; The most preferred combination being combined as two kinds of deoxy purine glycosides and other nucleoside a kind of or base, realizes following goal of the invention:

Deoxy purine nucleoside and other nucleoside or base are human normal nucleoside, with deoxy purine nucleoside for principal agent, the combination being accessory drugs with other nucleoside, base is for the preparation of the medicine of the multiple common cancers such as treatment gastric cancer, pulmonary carcinoma, hepatocarcinoma, colon cancer, the esophageal carcinoma, cancer of pancreas, breast carcinoma, genital system tumor, and therapeutic effect is remarkable.

For solving above technical problem, technical scheme of the present invention is:

The antitumor drug that deoxy purine nucleoside is prepared with other nucleoside or base composition, described medicine is combined by any one in deoxyadenosine and deoxyguanosine and other nucleoside and makes; Or combined by any one in deoxyadenosine and deoxyguanosine and base and make.

Below the further improvement to technique scheme:

Described nucleoside is guanosine.

Described nucleoside is guanine.

Described nucleoside is adenosine.

Described deoxyadenosine, guanosine, deoxyguanosine molar concentration rate are 1-3:0.5-2:1.

Described deoxyadenosine, guanosine, deoxyguanosine molar concentration rate are 1:0.5:1.

Described deoxyadenosine, guanosine, deoxyguanosine molar concentration rate are 3:2:1.

Described deoxyadenosine, guanosine, deoxyguanosine molar concentration rate are 1:1:1.

Described deoxyadenosine, guanine, deoxyguanosine molar concentration rate are: 1:1:1.

Described deoxyadenosine, guanine, deoxyguanosine molar concentration rate are: 1:0.5:1.

Described deoxyadenosine, adenosine, deoxyguanosine molar concentration rate are: 1:1:1.

Described deoxyadenosine, adenosine, deoxyguanosine molar concentration rate are: 1:0.5:1.

Described medicine makes injection or tablet or slow releasing agent.

An application for the antitumor drug that deoxy purine nucleoside is prepared with other nucleoside or base composition, described injection, it consists of:

Deoxyadenosine 14-16g

Guanosine 7.6-8.6g

Deoxyguanosine 15.9-16.9g

Injection soybean oil 190-210g

Injection lecithin 10-12g

Glycerol for injection 21-23g

Water for injection adds to 2000ml.

Described injection, it consists of:

Deoxyadenosine 15.5g

Guanosine 8.1g

Deoxyguanosine 16.4g

Injection soybean oil 200g

Injection lecithin 11g

Glycerol for injection 22g

Water for injection adds to 2000ml.

Described tablet, it consists of:

Deoxyadenosine 7.65-7.85g

Guanosine 3.95-4.15g

Deoxyguanosine 8.1-8.3g

Microcrystalline Cellulose 7.4-7.6g

Starch 5.9-6.1g

Magnesium stearate 1.4-1.6g

Described tablet is the amount of 100 tablets of tablets.

Described tablet, it consists of:

Deoxyadenosine 7.75g

Guanosine 4.05g

Deoxyguanosine 8.2g

Microcrystalline Cellulose 7.5g

Starch 6g

Magnesium stearate 1.5g

Described tablet is the amount of 100 tablets of tablets.

A preparation method for the antitumor drug that deoxy purine nucleoside is prepared with other nucleoside or base composition, is characterized in that: the preparation of described injection comprises the following steps:

The outfit of a, crude drug;

The preparation of b, oil phase;

The preparation of c, aqueous phase;

The preparation of d, colostrum;

E, tune pH;

F, breast are even, filtration;

J, Quan Jian, packaging, warehouse-in.

Described tune pH step, adjusts pH to 6-8.

Described breast is even, filtration step, Emulsion homogenate 20 times under 100 mPa pressure, and emulsion is with 0.45 μm of filtering with microporous membrane, and by Emulsion embedding in cleaned injection ampoule, logical nitrogen, sealing by fusing, puts pressure sterilizing 20 min under 121 DEG C of conditions.

Adopt described method, the obtained fat emulsion for injection of particle diameter between 100 ~ 500nm.

The preparation method of described tablet, comprises the following steps:

The outfit of a, crude drug;

B, raw material sieve respectively;

C, mix, sieve;

D, starch sieve;

E, add magnesium stearate;

F, tabletting, Quan Jian, packaging, warehouse-in.

Described raw material sieves respectively: take deoxyadenosine, guanosine, deoxyguanosine, starch, magnesium stearate and cross 100 mesh sieves respectively, microcrystalline Cellulose crosses 60 mesh sieves.

Described mixing, sifting step, mix deoxyadenosine, guanosine, deoxyguanosine with the equivalent method of progressively increasing with microcrystalline Cellulose, fully mix three times after 40 mesh sieves.

Described medicine, the IC50 value of the IC50 value to different tumor cell: Hela is the IC50 value of 0.6mmol/L, Bel-7402 is 0.9mmol/L; Suppression ratio to different tumor cell effect 48h: Hela suppression ratio is 88.43%; 7402 suppression ratio are 77.21%; HepG2 suppression ratio is 77.45%; SPC suppression ratio is 72.04%.

Compared with prior art, its advantage is in the present invention:

1, all nucleoside that the present invention relates to, base are the koinomatter in animal body, are the necessary nutrient substance of human normal tissue cell's growth and breeding.The discovery of human normal deoxyadenosine and nucleoside or base composition anti-cancer agent fundamentally changes the research direction of anti-cancer agent, discloses the new rule of oncotherapy;

2, deoxy purine nucleoside and other nucleoside or base composition are for the preparation of the medicine for the treatment of the multiple common cancers such as gastric cancer, pulmonary carcinoma, hepatocarcinoma, colon cancer, breast carcinoma, the esophageal carcinoma, cancer of pancreas, genital system tumor, antitumous effect is obvious, has the feature of wide spectrum, low toxicity; The IC50 value of the IC50 value to different tumor cell: Hela is the IC50 value of 0.6mmol/L, Bel-7402 is 0.9mmol/L; Suppression ratio to different tumor cell effect 48h: Hela suppression ratio is 88.43%; 7402 suppression ratio are 77.21%; HepG2 suppression ratio is 77.45%; SPC suppression ratio is 72.04%;

3, this antitumor drug is to the acellular toxic action of normal cell, is the great advantage of this invention;

4, deoxy purine nucleoside and other nucleoside or base composition antitumor action multiplicity increase, and significantly increase druggability; To the pharmacodynamics test of CT-26 tumor-bearing mice, adenine low dose group tumour inhibiting rate=-25.85%, adenine high dose group tumour inhibiting rate=51.03%, combination low dose group tumour inhibiting rate=54.01%, combination high dose group tumour inhibiting rate=64.93%;

5, deoxynucleoside and other nucleoside, base composition medication can delay the generation of drug resistance of tumor; Tumor inhibition mice multidrug resistance gene detects, animals administer 16 days, and conventional chemotherapy group multidrug resistance gene obviously raises, and drug resistance appears in chemotherapy; Adopt this composition of medicine group Multidrug Resistance Gene Expression without significant change, do not have drug resistance to occur.

accompanying drawing illustrates:

In Fig. 1: combination anti-tumor experiment DNMT gene test, remove genomic DNA reaction schematic diagram;

In Fig. 2: combination anti-tumor experiment DNMT gene test, reverse transcription reaction schematic diagram;

Fig. 3: the change schematic diagram of tumor inhibition small mouse body weight;

Fig. 4: during tumor inhibition mice multidrug resistance gene detects, removes genomic DNA reaction schematic diagram;

Fig. 5: during tumor inhibition mice multidrug resistance gene detects, reverse transcription reaction schematic diagram.

Detailed description of the invention

Below the preferred embodiments of the present invention are described, should be appreciated that preferred embodiment described herein is only for instruction and explanation of the present invention, is not intended to limit the present invention.

Following various material or its combination represent with digital code respectively in an embodiment, table specific as follows:

embodiment 1

245 combinations, 25 combinations, 2145 combination inhibiting tumor assay, specific experiment method is as follows:

1, the inoculation of cell

Get the Hela cell, 7402 cells that are in exponential phase, the living cells suspension of (2.0 ~ 3.0) × 104/mL is diluted to trypsinization, be inoculated in 2 ~ 10 row in 96 well culture plates, often row establish 6 multiple holes, every hole 100 μ L, 11st row add the RPMI-1640 culture fluid of 100 μ L containing 10% hyclone, and periphery is filled with 100 μ L PBS, is placed in 37 DEG C, 5%CO ₂24h is cultivated in incubator.

2, dosing intervention

After cell attachment, discard culture fluid in hole, 245 combinations are added successively by order from big to small, 2, the mol ratio of 4,5 is 2:1:2, seven Concentraton gradient (C1 ~ C7) of composition of medicine, its total concentration is respectively 2.0 mmol/L, 1.0 mmol/L, 0.5 mmol/L, 0.25 mmol/L, 0.125 mmol/L, 0.0625 mmol/L, 0.03125mmol/L;

25 combinations, the mol ratio of 2 and 5 is 1:1; Seven Concentraton gradient (C1 ~ C7) are the same;

2145 combinations, the mol ratio of 2,1,4,5 is 2:1:1:2, and seven Concentraton gradient (C1 ~ C7) are the same;

With to make up a prescription containing the RPMI-1640 culture fluid of 10% hyclone and doubling dilution is to desired concn successively, every hole adds 200 μ L;

Blank group, negative control group add the RPMI-1640 culture fluid 200 μ L containing 10% hyclone respectively, be placed in 37 DEG C, the incubator of 5%CO2 cultivates.

3, colour generation

Cultivate after 48h, observe tumor cell form under drug effect, quantity change and take pictures, add the MTT 20 μ L of 5mg/mL, after continuing to cultivate 4h, careful suction abandons whole supernatant, and every hole adds the DMSO of 150 μ L, and slight concussion is dissolved completely to crystalline particle.

4, colorimetric

Each hole optical density value (OD) is measured with 570nm and 630nm place by microplate reader.

5, inhibitory rate of cell growth is calculated

Be calculated as follows the suppression ratio of drug on tumor cell: tumour inhibiting rate (%)=(1-medication group OD570 meansigma methods/matched group OD570 meansigma methods) × 100%.

6, IC50 is calculated

According to formulae discovery: IC50=lg-1 [Xm-i (Σ P-0.5)], repeat 4 times, average;

Wherein: Xm: the logarithm value of the Cmax of design;

I: the logarithm value of each concentration multiple proportions concentration;

Σ P: each group growth inhibition ratio sum;

0.5: empirical.

Table 1 25 combination, 245 combinations and IC50 (mmol/L) values of 2145 kinds of combinations to Hela, Bel-7402

Found out as seen by table 1,245 combinations after are preferably better than 25 combinations and 2145 kinds of combinations (illustrating: the less antitumor action of IC50 numerical value is stronger).

Embodiment 2

The Factorial Design experiment of wantonly three kinds of external tumor suppressions of nucleoside combinations, specific experiment method is as follows:

1, the inoculation of cell

See " inoculation of cell " step in embodiment 1

2, dosing intervention

After cell attachment, discard culture fluid in hole, add concentration from front to back successively and be the combination (123,124,125,126,134,135,136,145,146,156,234,235,236,245,246,256,345,346,356,456) that 1 (adenosine) of 3.7mmol/L, 2 (deoxyadenosines), 3 (adenine), 4 (guanosines), 5 (deoxyguanosines), 6 (guanine) six medicines and this six medicine mol ratios are 1:1:1;

Make up a prescription to desired concn with the RPMI-1640 culture fluid containing 10% hyclone, every hole adds 200 μ L;

Blank group, negative control group add the RPMI-1640 culture fluid 200 μ L containing 10% hyclone respectively, are placed in 37 DEG C, 5%CO ₂incubator in cultivate.

3, colour generation

4, colorimetric

5, inhibitory rate of cell growth is calculated

6, experimental result:

Application mtt assay measures the inhibiting tumor assay of six kinds of medicines to Hela cell, 7402 cells, HepG2 cell and SPC cell, specifically in table 2:

Table 2 six kinds of medicines and the suppression ratio (%) of combination to different tumor cell effect 48h thereof

As can be seen from Table 2:

1, the tumor-inhibiting action that the tumor-inhibiting action of drug regimen is more independent than medicine is more remarkable;

2, the most remarkable with the tumor-inhibiting action of deoxyadenosine-guanosine-deoxyguanosine combination (245 combination) in these 20 combinations, 88.4%, 74.3%, 77.5%, 72.0% is reached respectively to the tumor-inhibiting action of Hela cell, 7402 cells, HepG2 cell and SPC cell.

Experiment conclusion:

1, combine antitumor action under same concentration conditions and be better than monomer;

2, deoxyadenosine-guanosine-deoxyguanosine combination antitumor action is the strongest; What antitumor action took second place is deoxyadenosine-deoxyguanosine-guanine, adenosine-deoxyadenosine-deoxyguanosine.

Embodiment 3

The orthogonal of deoxyadenosine-guanosine-deoxyguanosine (245) combined concentration proportioning, specific experiment step is as follows:

1, the inoculation of cell

2, dosing intervention

After cell attachment, discard culture fluid in hole, dosing regimen is as follows: 2 (deoxyadenosines), 4 (guanosines), 5 (deoxyguanosine) concentration are 3mmol/L, designs obtain 9 groups and is respectively: the mol ratio that 2 (deoxyadenosines), 4 (guanosines), 5 (deoxyguanosines) add is respectively: (1) 1:1:1; (2) 2:1:2; (3) 3.25:1:3.25; (4) 1:2:2; (5) 1:1:1.46(6) 3:2:1; (7) 1:3.25:3.25; (8) 2:3:1; (9) 1.55:1.55:1;

The step of concrete dosing:

(1) 2,4,5 volumes added respectively are 67 μ L, 67 μ L, 67 μ L;

(2) 2,4,5 volumes added respectively are 80.4 μ L, 40.2 μ L, 80.4 μ L;

(3) 2,4,5 volumes added respectively are 87.1 μ L, 26.8 μ L, 87.1 μ L;

(4) 2,4,5 volumes added respectively are 40.2 μ L, 80.4 μ L, 80.4 μ L;

(5) 2,4,5 volumes added respectively are 58.1 μ L, 58.1 μ L, 84.8 μ L;

(6) 2,4,5 volumes added respectively are 100.5 μ L, 67 μ L, 33.5 μ L;

(7) 2,4,5 volumes added respectively are 26.8 μ L, 87.1 μ L, 87.1 μ L;

(8) 2,4,5 volumes added respectively are 67 μ L, 100.5 μ L, 33.5 μ L;

(9) 2,4,5 volumes added respectively are 75.9 μ L, 75.9 μ L, 49.2 μ L;

Make up a prescription to desired concn with the RPMI-1640 culture fluid containing 10% hyclone, every hole total amount is 200 μ L;

3, colour generation

4, colorimetric

5, inhibitory rate of cell growth is calculated

Experimental result is in table 3:

The tumour inhibiting rate (%) of table 3 245 proportioning combination to different tumor cell effect 48h

As seen from Table 3:

1, when deoxyadenosine-guanosine-deoxyguanosine molar concentration rate is 2:1:2, antitumor action is best, and taking second place is deoxyadenosine-guanosine-deoxyguanosine 3:2:1, again deoxyadenosine-guanosine-deoxyguanosine 1:1:1.

Experiment conclusion:

Application mtt assay measures medicine and obtains Hela cell, 7402 cell inhibiting tumor assay: when 2,4, No. 5 drug ratios are 2:1:2, tumor-inhibiting action is the most remarkable.

Embodiment 4:

The outer metabolism of deoxyadenosine-guanosine-deoxyguanosine assembly, specific experiment method is as follows:

1, the inoculation of cell

Get the Hela cell, 7402 cells that are in exponential phase, be diluted to the living cells suspension of (1.0 ~ 2.0) × 105/mL with trypsinization, be inoculated in 6 well culture plates, every hole 2.5mL.Be placed in 37 DEG C, 5%CO2 incubator cultivates 24h.

2, dosing intervention

After cell attachment, discard culture fluid in hole, each cell adds molar concentration successively, and to be 2 (deoxyadenosines) of 3.7mmol/L, 4 (guanosines), 5 (deoxyguanosines) and 245 combination matchings be mol ratio 2:1:2, every hole 2.5mL, is placed in 37 DEG C, the incubator of 5%CO2 cultivates.

3, detect

After cultivation 24h, 48h, get cell conditioned medium liquid, 0.22 μm of frit, efficient Liquid Detection.

4, testing conditions:

Equipment: U.S. Dai An company U3000 high-efficient liquid instrument

Liquid-phase condition

Chromatographic column: C18AQ, 250 × 4.6mm × 5 μm

Column temperature: 30 DEG C

Mobile phase: water (phosphoric acid adjusts PH to be 6.6): methanol=90:10

Flow velocity: 1ml/min

Wavelength: 254nm.

Specific experiment the results are shown in Table 4, table 5:

Table 4 deoxyadenosine-guanosine-deoxyguanosine is combined in the metabolic condition (under HPLC peak area) in Hela

	0	24 hours	48 hours
				Deoxyadenosine	655	518	371
Guanosine	294	287	210
				Deoxyguanosine	300	268	259

Table 5 deoxyadenosine-guanosine-deoxyguanosine is combined in the metabolic condition (under HPLC peak area) in Bel-7402

	0	24 hours	48 hours
				Deoxyadenosine	652	564	414
Guanosine	291	286	233
				Deoxyguanosine	298	295	292

From in table 4, table 5: nucleoside combinations was through 24,48 hours and tumor cell culture, and in culture medium, concentration is basicly stable.

Experiment prompting: experiment in vitro data are caused by composition.In addition, prompting preparation tumor new drug has feasibility.

Embodiment 5:

Deoxyadenosine-guanosine-deoxyguanosine combination preparation fat emulsion for injection and preparation method thereof:

Deoxyadenosine-guanosine-deoxyguanosine combination preparation fat emulsion for injection, it consists of:

Deoxyadenosine 15.5g

Guanosine 8.1g

Deoxyguanosine 16.4g

Injection soybean oil 200g

Injection lecithin 11g

Glycerol for injection 22g

Water for injection adds to 2000ml

The preparation method of deoxyadenosine-guanosine-deoxyguanosine combination preparation fat emulsion for injection, comprises the following steps:

The outfit of a, crude drug

Various crude drug is equipped with according to following ratio:

Deoxyadenosine 15.5g

Guanosine 8.1g

Deoxyguanosine 16.4g

Injection soybean oil 200g

Injection lecithin 11g

Glycerol for injection 22g

The preparation of b, oil phase

Recipe quantity injection soybean oil is put in container, stirs, make oil phase;

The preparation of c, aqueous phase

Recipe quantity deoxyadenosine, guanosine, deoxyguanosine, injection lecithin, glycerol for injection and appropriate water for injection are joined in container, stirs, make aqueous phase;

The preparation of d, colostrum

The oil phase obtained by step b under agitation adds in the aqueous phase that step c obtains, and high speed shear makes colostrum;

E, tune pH

Adjust pH to 6-8;

F, breast are even, filtration

Emulsion homogenate 20 times under 100 mPa pressure, emulsion with 0.45 μm of filtering with microporous membrane, by Emulsion embedding in cleaned injection ampoule, logical nitrogen, sealing by fusing, puts pressure sterilizing 20 min under 121 DEG C of conditions, obtains the fat emulsion for injection of particle diameter between 100 ~ 500nm;

J, Quan Jian, packaging, warehouse-in.

This product preparation process advanced person is feasible, and drug loading is comparatively large, also has slow release, targeting simultaneously, provides the effects such as energy for body.

Embodiment 6:

Deoxyadenosine-guanosine-deoxyguanosine combination preparation tablet and preparation method thereof:

Deoxyadenosine-guanosine-deoxyguanosine combination preparation tablet, it consists of:

Deoxyadenosine 7.75g

Guanosine 4.05g

Deoxyguanosine 8.2g

Microcrystalline Cellulose 7.5g

Starch 6g

Magnesium stearate 1.5g

Make 100

Deoxyadenosine-guanosine-deoxyguanosine combination preparation method for preparing tablet thereof, comprises the following steps:

The outfit of a, crude drug:

Various crude drug is equipped with according to following ratio:

Deoxyadenosine 7.75g

Guanosine 4.05g

Deoxyguanosine 8.2g

Microcrystalline Cellulose 7.5g

Starch 6g

Magnesium stearate 1.5g

B, raw material sieve respectively:

Take recipe quantity deoxyadenosine, guanosine, deoxyguanosine, starch, magnesium stearate and cross 100 mesh sieves respectively, it is for subsequent use that microcrystalline Cellulose crosses 60 mesh sieves;

C, mix, sieve:

Deoxyadenosine, guanosine, deoxyguanosine are mixed with the equivalent method of progressively increasing with microcrystalline Cellulose, fully mixes three times after 40 mesh sieves;

D, starch sieve

Material in starch and step c is crossed 40 mesh sieves and fully mixes three times;

E, add magnesium stearate

Above-mentioned material is added recipe quantity magnesium stearate mix homogeneously, cross 20 mesh sieves;

F, tabletting, Quan Jian, packaging, warehouse-in.

Illustrate:

1, above concrete dosage is median, can have floating of upper and lower 50-100%;

2, the adjuvant (soybean oil, cellulose, starch etc.) of medicine according to practical situation and can require adjustment.

Embodiment 7

Combination anti-tumor experiment DNMT gene test:

One, experiment material

1, reagent

RNAiso extracts test kit; Chloroform; Isopropyl alcohol; DEPC water; PrimeScriptTMRT reagent Kit with gDNA Eraser(Perfect Real Time); SYBR Premix Ex TaqTM(Tli RNaseH Plus).

2, instrument:

Quantitative PCR apparatus (eppendorf company); SW-CJ-2F type clean work station (Purifying Equipment Co., Ltd., Suzhou); Anke TGL-16B centrifuge (Anting Scientific Instrument Factory, Shanghai); Constant-temperature metal bath (Hangzhou BIOER Technology Co., Ltd); NV3000 Micro-Spectrophotometer(vastech); Pipettor (scope 100-1000 microlitre, 10-100 microlitre, 0.5-10 microlitre) (eppendorf company).

Two, primer sequence

House-keeping gene (GAPDH) primer sequence

H-GAPDH-F：5'-TGGGTGTGAACCATGAGAAGT-3' 57.4

H-GAPDH-R：5'-TGAGTCCTTCCACGATACCAA-3' 57.9

Genes of interest primer sequence

H-DNMT1-F: 5'-CCCTGAGCCCTACCGAATTG-3' 57.7

H-DNMT1-R: 5'-GTAGCTCGCTGGAGTGGACTTG-3' 59.9

Three, sample treatment

1, the thawing and cultivation of cell

From liquid nitrogen container, take out human cervical carcinoma Hela cell, people's liver cancer cell line 7402,37 DEG C of water-baths thaw rapidly.After about 1min, frozen liquid in pipe dissolves completely, put into superclean bench again, with suction pipe sucking-off cell suspension, to inject in culture bottle and to add 10 times of RPMI-1640 culture fluid (containing 10% hyclone, penicillin 100U/mL and streptomycin 100 μ g/mL), being placed in 37 DEG C, 5%CO ₂cultivate in incubator, change a culture fluid next day, cell is monolayer adherence growth, within every 2 ~ 3 days, goes down to posterity 1 time.

2, dosing intervention

(1) inoculation of cell

Get the Hela cell, 7402 cells that are in exponential phase, be diluted to the living cells suspension of (4.0 ~ 6.0) × 104/mL with trypsinization, be inoculated in 25cm ²culture bottle in, every bottle of 5 mL, 2 bottles, each cell.Be placed in 37 DEG C, 5%CO ₂24h is cultivated in incubator.

(2) dosing intervention

After cell attachment, discard culture in glassware liquid, Hela cell, 7402 cells are divided into three groups, and one group adds 245 combination 2mmol/L, 10 mL, and one group adds 245 combination 1mmol/L, 10 mL, and another group adds the RPMI-1640 culture fluid 10mL containing 10% hyclone.Administration time 48h.

Four, RNA extracting

1, cracking collecting cell

(1) pour out culture fluid, clean once with 1*PBS.

(2) add the RNAiso Plus of 1mL in every bottle of cell, horizontal positioned for a moment, makes lysate be uniformly distributed in cell surface and cell lysis, then uses liquid-transfering gun piping and druming cell to make it come off.

(3) interior celliferous lysate is transferred in centrifuge tube, with liquid-transfering gun repeatedly pressure-vaccum until without obvious sediment in lysate.

(4) room temperature leaves standstill 5min.

2, in above-mentioned homogenate lysate, add chloroform (1/5 volume of RNAiso Plus), cover tightly centrifuge tube lid, low, volatile with hands concuss 15s(chloroform boiling point, during vibration, care should be used to centrifuge tube lid flicks suddenly).After solution fully emulsified (without noted phase separation phenomena), then room temperature leaves standstill 5min.

3,12000g 4 DEG C of centrifugal 15min.

4, from centrifuge, carefully take out centrifuge tube, now homogenate is divided into three layers, that is: colourless supernatant, middle white egg lamina albae and be with coloured lower floor organic facies.Aspirate supernatant is transferred in new centrifuge tube and (must guard against sucking-off white intermediate layer).

5, in supernatant, add isopyknic isopropyl alcohol, after the centrifuge tube that turns upside down fully mixes, at 15 ~ 30 DEG C, leave standstill 10min.

6,12000g 4 DEG C of centrifugal 10min.Generally after centrifugation, there will be precipitation bottom test tube.

7, the cleaning of RNA precipitation

Careful supernatant discarded, add 75% room temperature ethanol 1mL(along centrifugal tube wall to be lentamente sure not to touch precipitation), turn upside down washing centrifuge tube tube wall gently, carefully discards ethanol (in order to the salt ion content in control RNA better, Ex-all ethanol of should trying one's best) after 12000g 4 DEG C of centrifugal 5min.

8, the dissolving of RNA

Drying at room temperature precipitation 2 ~ 5min(cannot centrifugal or heat drying, otherwise RNA will be difficult to dissolve), add appropriate RNase-free water dissolution precipitation, precipitation can be blown and beaten gently if desired, with NV3000 Micro-Spectrophotometer detection and in-80 DEG C of preservations after RNA precipitation is dissolved completely with liquid-transfering gun.

9, RNA testing result

	260/230	260/280	ng/μl
				Hela-matched group	2.2215	1.8423	1385.5
Hela-1mmol/L	2.1203	1.8351	2532.4
				Hela-2mmol/L	2.1567	1.8667	2783.6
7402-matched group	2.1651	1.8775	1326.2
				7402-1mmol/L	2.1871	1.856	1702.4
7402-2mmol/L	2.1952	1.8565	1796.7

Five, reverse transcription

1, genomic DNA reaction is removed

As Fig. 1, by following composition in preparing reaction mixture on ice, in order to ensure the accuracy that reactant liquor is prepared, when carrying out every reaction, first should press the amount preparation Master Mix of stoichiometric number+2, and then being dispensed in each reaction tube, finally adding RNA sample.

2, reverse transcription reaction

As Fig. 2, reactant liquor preparation is please carried out on ice.In order to ensure the accuracy that reactant liquor is prepared, when carrying out every reaction, first should press the amount preparation Master Mix of stoichiometric number+2, and then subpackage 10 μ l is in each reaction tube.Reverse transcription reaction is carried out immediately after soft mixing.

Six, fluorescence quantitative PCR detection

1, experiment sample

Using cDNA diluted sample 10 doubly as machine testing in template.

2, PCR reactions steps

(1) reaction mixture is prepared

(2) PCR cycling condition

(3) operation of instrument

After completing above-mentioned steps, 8 unions having added sample are placed in quantitative PCR apparatus (eppendorf company) and react.

Seven, relative quantification result

DNMT1 gene relative quantification result

Experiment proves: nucleoside combinations, by raising DNMT1, makes oncogene be in hyper-methylation state, thus Tumor suppression gene expression, thus Tumor suppression growth.

Embodiment 8

The pharmacodynamic study of combination to CT-26 tumor-bearing mice

1. experiment material

Tumor strain systematic name: CT26.WT Chinese: mouse colonic cell

Half and half, 6 week age of laboratory animal BALB/c mouse 72 male and female

2. the preparation of test sample, reference substance and medication thereof

(1) preparation of test sample

Test sample administration preparation on the same day in an aseptic environment, test sample tag identifier after preparation, adenine low dose group (AL) adopts blue label, adenine high dose group (AH), combination low dose group (ZL), combination high dose group (ZH) to adopt red-ticket, fills in the information such as test sample numbering, preparation date, compound concentration and preparation people on label simultaneously.Compound method according to the form below carries out.

Table 6, medicine ordinance method abridged table

Points for attention: after test sample boils dissolving, need be cooled to room temperature (about 25 DEG C) standardize solution, filtration, high pressure afterwards, room temperature preservation; Test sample preparation is rear and daily terminate rear shading stored refrigerated.

(2) preparation of positive reference substance

Get 1 5-FU injection, with normal saline mixing, the ultimate density of 5-FU injection is 7.5mg/ml, and test sample preparation is rear and daily terminate rear shading stored refrigerated.

(3) route of administration: all adopt intraperitoneal administration

(4) dosage: medicine low dose group: 500mg/kg/d; Medicine high dose group: 1000 mg/kg/d; Positive controls: 15mg/kg/d

(5) administration frequency and the cycle

Administration group and normal saline group every day at upper and lower noon each administration 1 time, positive controls once a day.Each group of equal successive administration 14 days.

3, experimental technique

(1) cell recovery, cultivation

Conventional recovery mouse junction cancer CT-26 cell strain, being cultured to cell, to be in the multiple rise period for subsequent use.

(2) divide into groups, set up mice with tumor model

Be divided into 6 groups at random, adenine low dose group, adenine high dose group by all mices of the large young pathbreaker of body weight, combination low dose group, combination high dose group, normal saline group and positive controls, often organize 12.It is 5 × 106/ml that cell good for collection growth conditions is adjusted cell concentration, implants in right side of mice armpit subcutaneous tissue with syringe, every only 0.2 ml.

(3) animals administer

Each group all adopts intraperitoneal administration, and administration group upper and lower noon each administration 1 time, normal saline group gives equal-volume sodium chloride injection.Positive controls gives 5-FU, normal saline group and positive controls administration in evening every day 1 time.Each group of equal successive administration 14 days.

4, the observation of index

(1) tumor is observed: during off-test, puts to death animal and dissects, and observes the situation of tumor at when dissected.

(2) measurement of tumor: after complete for tumor tissues taking-up, dry bloodstain, weigh.

(3) tumour inhibiting rate is calculated

Tumour inhibiting rate=(1-experimental group tumor weight/matched group tumor weight) × 100%

5, date processing

(1) adopt Relative tumor appreciation rate T/C (%) as test evaluation index

The computing formula of gross tumor volume (TV):

V=1/2×a×b2，

Wherein a, b represent the length of tumor body and wide respectively;

The computing formula of relative tumour volume (RTV): RTV=Vt/V0,

Wherein V0 is that when dividing cage administration, (d0) measures gained gross tumor volume,

Vt is gross tumor volume when measuring each time;

Relative tumor appreciation rate T/C(%) computing formula: T/C%=TRTV/CRTV × 100%

Wherein: TRTV represents treatment group RTV,

CRTV represents negative control group RTV;

In principle, evaluation criterion is: T/C(%) > 40% is for invalid; T/C(%)≤40%, and be effective through statistical procedures P < 0.05.

(2) curative effect of test sample is done in order to tumor-like hyperplasia as evaluation index

The computing formula of tumor-like hyperplasia: tumor-like hyperplasia %=(1-T/C) × 100%.

(3) according to by EXCEL computed in software, average, standard deviation are gone out to mouse tumor tuple, represent with (mean value ± SD) in report; Carry out pairing T-and check (test sample group and matched group compare).

6. experimental result

(1) tumor is observed

5-Fu group, adenine high dose group and combination high dose group all occur that tumor is less, and peplos is complete, demarcates obviously with surrounding tissue.Normal saline group tumor is comparatively large, and Partial tumors tissue has necrosis, has adhesion with surrounding tissue.

(2) tumor weight

The tumor weight after medication respectively organized by table 7

Grouping	Number of cases	Tumor heavy (g)
			NS group	12	3.288±0.943
AL group	12	4.138±1.001
			AH group	11	1.610±0.775
ZL group	12	1.512±1.021
			ZH group	12	1.153±0.9257
5-Fu group	11	1.497±0.9868

When 5-FU group, high dose group administration the 13rd day, 1 mice occurs dead.

(3) tumour inhibiting rate

Adenine low dose group tumour inhibiting rate=-25.85%

Adenine high dose group tumour inhibiting rate=51.03%

Combination low dose group tumour inhibiting rate=54.01%

Combination high dose group tumour inhibiting rate=64.93%

Tumor inhibition proves: when total concentration is identical, and " combination " tumor suppression obviously strengthens; Now, single concentration declines 3/5 and 4/5 respectively; With regard to single medicine, external tumor-inhibiting action obviously increases.

Embodiment 9: tumor inhibition mice animation contrasts

1, the observation, date processing etc. of experiment material, compounding medicine and medication thereof, experimental technique, index, with embodiment 8.

2, observation index

(1) body weight of animal.

All animals were weighed before lotus tumor, all weighed before each administration.

(2) animal feed drinking-water situation.

Observe and record animal feed and drinking-water situation.

(3) animal general state.

Observation experiment animal spirit, active state etc.

3, experimental result

(1) change of body weight

Medication the 5th day, it is slack-off that 5-Fu treated animal starts to occur that body weight increases, and the 8th day body is reppeared and now fallen.There is weight loss on 10th day in adenine high dose group animal, the decline of body weight appears on the 10th day in combination high dose group in medication.All the other each treated animals all present the rising of body weight.At the end of medication, each treated animal weight ratio is comparatively, 5-Fu group, adenine high dose group, combination high dose group, comparatively matched group all has obvious decline (P<0.05), and all the other are respectively organized comparatively matched group and compare, body weight without significant change, as shown in Figure 3.

(2) feed, drinking-water situation

Medication the 3rd day, the feed of 5-Fu treated animal started to reduce, and amount of drinking water, at the 6th day, starts to occur reducing.Adenine high dose group and combination high dose group all occur that at the 8th day feed and amount of drinking water reduce.All the other each group started at the 12nd day, and food-intake starts to increase.

(3) animal general state

Medication the 8th day, 5-Fu treated animal started appearance activity and reduces, and trichosis setosa, becomes thin.Adenine high dose group and combination high dose group were at the 10th day, and appearance activity reduces, and becomes thin.All the other each treated animals are without exception.Illustrate that the comprehensive toxicity of " combination " is lower than 5-Fu and adenine list medicine.

Embodiment 10:

Tumor inhibition mice multidrug resistance gene detects:

One, the foundation of mouse-borne tumor model and dosing intervention

1, cell recovery, cultivation

2, divide into groups, set up mice with tumor model

Be divided into 3 groups at random, drug regimen group, normal saline group and positive controls by all mices of the large young pathbreaker of body weight, often organize 8.It is 5 × 106/ml that cell good for collection growth conditions is adjusted cell concentration, implants in right side of mice armpit subcutaneous tissue with syringe, every only 0.2 ml.

3, animals administer

Each group all adopts intraperitoneal administration, and drug regimen group dosage is 1000mg/kg/d, each administration of upper and lower noon 1 time.Normal saline group gives equal-volume sodium chloride injection, administration in evening every day 1 time.Positive controls gives cisplatin 3mg/kg, 1 time weekly; Cyclophosphamide and each 3mg/kg of 5-FU, once a day.Each group of equal successive administration 16 days.

4, frozen tumor tissues

After medication terminates, take out tumor tissues, and put into liquid nitrogen freezing rapidly.

Two, RNA extracting

1, the RNA freezed by ultralow temperature is transferred to rapidly in the mortar with Liquid nitrogen precooler after extracting sample weighing, use Yan Zha tissue abrasion, constantly add liquid nitrogen therebetween, until be ground into powder (without obvious visible particle, if do not have to grind yield and the quality that thoroughly can affect RNA).

2, then in mortar, add appropriate RNAiso Plus, covered completely by the sample be ground into powder, then room temperature leaves standstill, until sample melts completely, then continues to be ground to the transparent shape of lysate with grinding toothed oak.

3, be transferred in centrifuge tube by homogenate, room temperature leaves standstill 5 minutes.

4,12,000 g 4 DEG C is centrifugal 5 minutes.

5, careful Aspirate supernatant, moves in new centrifuge tube and (is sure not to draw precipitation).

6, in above-mentioned even supernatant, add chloroform (1/5 volume of RNAiso Plus), cover tightly centrifuge tube lid, low, volatile with hands concuss 15s(chloroform boiling point, during vibration, care should be used to centrifuge tube lid flicks suddenly).After solution fully emulsified (without noted phase separation phenomena), then room temperature leaves standstill 5min.

7,12000g 4 DEG C of centrifugal 15min.

8, from centrifuge, carefully take out centrifuge tube, now homogenate is divided into three layers, that is: colourless supernatant, middle white egg lamina albae and be with coloured lower floor organic facies.Aspirate supernatant is transferred in new centrifuge tube and (must guard against sucking-off white intermediate layer).

9, in supernatant, add isopyknic isopropyl alcohol, after the centrifuge tube that turns upside down fully mixes, at 15 ~ 30 DEG C, leave standstill 10min.

10,12000g 4 DEG C of centrifugal 10min.Generally after centrifugation, there will be precipitation bottom test tube.

11, the cleaning of RNA precipitation

12, the dissolving of RNA

Three, reverse transcription

1, genomic DNA reaction is removed

As Fig. 4, by following composition in preparing reaction mixture on ice, in order to ensure the accuracy that reactant liquor is prepared, when carrying out every reaction, first should press the amount preparation Master Mix of stoichiometric number+2, and then being dispensed in each reaction tube, finally adding RNA sample.

2, reverse transcription reaction

Reactant liquor preparation is please carried out on ice.In order to ensure the accuracy that reactant liquor is prepared, when carrying out every reaction, first should press the amount preparation Master Mix of stoichiometric number+2, and then subpackage 10 μ l is in each reaction tube.Reverse transcription reaction is carried out immediately, as Fig. 5 after soft mixing.

four, fluorescence quantitative PCR detection

1, experiment sample

Using cDNA diluted sample 10 doubly as machine testing in template.

, PCR reactions steps

(1) reaction mixture is prepared

(2) PCR cycling condition

(3) operation of instrument

five, MDR1 relative quantification result

Experiment proves: animals administer 16 days, and conventional chemotherapy group multidrug resistance gene obviously raises, and drug resistance appears in chemotherapy; " combination " group Multidrug Resistance Gene Expression, without significant change, does not have drug resistance to occur.

Claims

1. the antitumor drug prepared with other nucleoside or base composition of deoxy purine nucleoside, is characterized in that: described medicine is combined by any one in deoxyadenosine and deoxyguanosine and other nucleoside and makes; Or combined by any one in deoxyadenosine and deoxyguanosine and base and make.

2. the antitumor drug prepared with other nucleoside or base composition of a kind of deoxy purine nucleoside as claimed in claim 1, is characterized in that: described nucleoside is guanosine.

3. the antitumor drug prepared with other nucleoside or base composition of a kind of deoxy purine nucleoside as claimed in claim 1, is characterized in that: described nucleoside is guanine.

4. the antitumor drug prepared with other nucleoside or base composition of a kind of deoxy purine nucleoside as claimed in claim 1, is characterized in that: described nucleoside is adenosine.

5. the antitumor drug prepared with other nucleoside or base composition of a kind of deoxy purine nucleoside as claimed in claim 1, is characterized in that: described deoxyadenosine, guanosine, deoxyguanosine molar concentration rate are 1-3:0.5-2:1.

6. the antitumor drug prepared with other nucleoside or base composition of a kind of deoxy purine nucleoside as claimed in claim 5, is characterized in that: described deoxyadenosine, guanosine, deoxyguanosine molar concentration rate are 1:0.5:1.

7. the antitumor drug prepared with other nucleoside or base composition of a kind of deoxy purine nucleoside as claimed in claim 5, is characterized in that: described deoxyadenosine, guanosine, deoxyguanosine molar concentration rate are 3:2:1.

8. the antitumor drug prepared with other nucleoside or base composition of a kind of deoxy purine nucleoside as claimed in claim 5, is characterized in that: described deoxyadenosine, guanosine, deoxyguanosine molar concentration rate are 1:1:1.

9. the antitumor drug prepared with other nucleoside or base composition of a kind of deoxy purine nucleoside as claimed in claim 1, is characterized in that: described deoxyadenosine, guanine, deoxyguanosine molar concentration rate are: 1:1:1.

10. the antitumor drug prepared with other nucleoside or base composition of a kind of deoxy purine nucleoside as claimed in claim 1, is characterized in that: described deoxyadenosine, guanine, deoxyguanosine molar concentration rate are: 1:0.5:1.

The antitumor drug that 11. a kind of deoxy purine nucleoside as claimed in claim 1 are prepared with other nucleoside or base composition, is characterized in that: described deoxyadenosine, adenosine, deoxyguanosine molar concentration rate are: 1:1:1.

The antitumor drug that 12. a kind of deoxy purine nucleoside as claimed in claim 1 are prepared with other nucleoside or base composition, is characterized in that: described deoxyadenosine, adenosine, deoxyguanosine molar concentration rate are: 1:0.5:1.

The application of the antitumor drug that 13. a kind of deoxy purine nucleoside as claimed in claim 1 are prepared with other nucleoside or base composition, is characterized in that: described medicine makes injection or tablet or slow releasing agent.

The application of the antitumor drug that 14. a kind of deoxy purine nucleoside as claimed in claim 13 are prepared with other nucleoside or base composition, it is characterized in that: described injection, it consists of:

Deoxyadenosine 14-16g

Guanosine 7.6-8.6g

Deoxyguanosine 15.9-16.9g

Injection soybean oil 190-210g

Injection lecithin 10-12g

Glycerol for injection 21-23g

Water for injection adds to 2000ml.

The application of the antitumor drug that 15. a kind of deoxy purine nucleoside as claimed in claim 13 are prepared with other nucleoside or base composition, it is characterized in that: described injection, it consists of:

Deoxyadenosine 15.5g

Guanosine 8.1g

Deoxyguanosine 16.4g

Injection soybean oil 200g

Injection lecithin 11g

Glycerol for injection 22g

Water for injection adds to 2000ml.

The application of the antitumor drug that 16. a kind of deoxy purine nucleoside as claimed in claim 13 are prepared with other nucleoside or base composition, it is characterized in that: described tablet, it consists of:

Deoxyadenosine 7.65-7.85g

Guanosine 3.95-4.15g

Deoxyguanosine 8.1-8.3g

Microcrystalline Cellulose 7.4-7.6g

Starch 5.9-6.1g

Magnesium stearate 1.4-1.6g

Described tablet is the amount of 100 tablets of tablets.

The application of the antitumor drug that 17. a kind of deoxy purine nucleoside as claimed in claim 13 are prepared with other nucleoside or base composition, it is characterized in that: described tablet, it consists of:

Deoxyadenosine 7.75g

Guanosine 4.05g

Deoxyguanosine 8.2g

Microcrystalline Cellulose 7.5g

Starch 6g

Magnesium stearate 1.5g

Described tablet is the amount of 100 tablets of tablets.

The preparation method of the antitumor drug that 18. a kind of deoxy purine nucleoside as claimed in claim 1 are prepared with other nucleoside or base composition, is characterized in that: the preparation of described injection comprises the following steps:

The outfit of a, crude drug;

The preparation of b, oil phase;

The preparation of c, aqueous phase;

The preparation of d, colostrum;

E, tune pH;

F, breast are even, filtration;

J, Quan Jian, packaging, warehouse-in.

The preparation method of the antitumor drug that 19. a kind of deoxy purine nucleoside as claimed in claim 17 are prepared with other nucleoside or base composition, is characterized in that: described tune pH step, adjusts pH to 6-8.

The preparation method of the antitumor drug that 20. a kind of deoxy purine nucleoside as claimed in claim 17 are prepared with other nucleoside or base composition, it is characterized in that: described breast is even, filtration step, Emulsion homogenate 20 times under 100 mPa pressure, emulsion is with 0.45 μm of filtering with microporous membrane, by Emulsion embedding in cleaned injection ampoule, logical nitrogen, sealing by fusing, puts pressure sterilizing 20 min under 121 DEG C of conditions.

The preparation method of the antitumor drug that 21. a kind of deoxy purine nucleoside as claimed in claim 17 are prepared with other nucleoside or base composition, is characterized in that: adopt described method, the obtained fat emulsion for injection of particle diameter between 100 ~ 500nm.

The preparation method of the antitumor drug that 22. a kind of deoxy purine nucleoside as claimed in claim 17 are prepared with other nucleoside or base composition, is characterized in that: the preparation method of described tablet, comprises the following steps:

The outfit of a, crude drug;

B, raw material sieve respectively;

C, mix, sieve;

D, starch sieve;

E, add magnesium stearate;

F, tabletting, Quan Jian, packaging, warehouse-in.

The preparation method of the antitumor drug that 23. a kind of deoxy purine nucleoside as claimed in claim 21 are prepared with other nucleoside or base composition, it is characterized in that: described raw material sieves respectively: take deoxyadenosine, guanosine, deoxyguanosine, starch, magnesium stearate and cross 100 mesh sieves respectively, microcrystalline Cellulose crosses 60 mesh sieves.

The preparation method of the antitumor drug that 24. a kind of deoxy purine nucleoside as claimed in claim 21 are prepared with other nucleoside or base composition, it is characterized in that: described mixing, sifting step, deoxyadenosine, guanosine, deoxyguanosine are mixed with the equivalent method of progressively increasing with microcrystalline Cellulose, fully mixes three times after 40 mesh sieves.

The antitumor drug that 25. a kind of deoxy purine nucleoside as claimed in claim 1 are prepared with other nucleoside or base composition, it is characterized in that: described medicine, the IC50 value of the IC50 value to different tumor cell: Hela is the IC50 value of 0.6mmol/L, Bel-7402 is 0.9mmol/L; Suppression ratio to different tumor cell effect 48h: Hela suppression ratio is 88.43%; 7402 suppression ratio are 77.21%; HepG2 suppression ratio is 77.45%; SPC suppression ratio is 72.04%.