CN1088094A - 8-chlorine adenosine antineoplastic is used - Google Patents

Abstract

8-chlorine adenosine structural formula is as follows:

We at first adopt inside and outside medical science pharmacological experiment method to prove that 8-chlorine adenosine has the therapeutical effect of growth inhibited or the prolongation of host's life to various human and mammal malignant tumor, its anti-tumor activity is greater than 8-Cl-cAMP, toxicity can be made into multiple pharmaceutical dosage form (injection, tablet, capsule, syrup and other oral liquids, suppository etc.) and is used for oncotherapy far below known chemotherapeutic.

Description

8-chlorine adenosine antineoplastic is used

The invention belongs to the new medical application claim of known compound.

Background of invention: we are at research known compound 8-chlorine adenosine-3 ' 5 ' cycli phosphate, three fat (8-Cl-cAMP) (Free, C.A.and Paik, V.S.Endocrinology, 100: 1287-1293,1977) confirm in the antitumaous effect, the enzymatic hydrolysate 8-chlorine adenosine of 8-Cl-cAMP also has tangible anti-tumor activity and is better than 8-Cl-cAMP, we have at first synthesized 8-chlorine adenosine in China, and at first adopt in the body and external medical science pharmacological method, systematically studied the anti-tumor activity of 8-chlorine adenosine, proved our synthetic 8-chlorine adenosine leukaemia HL-60 and K the people ₅₆₂Cell line, mouse leukemia L ₁₂₁₀Cell (ascitic type), people's gastric cancer MGc80-3 cell and rat liver cancer solid tumor H ₂₂All reach the requirement of ministry of Health of China new drug approval rules etc. the therapeutic effect of multiple malignant tumor, proved the using value of 8-chlorine adenosine,, find the document relevant with this research novelty through Patent Office of the People's Republic of China's update search as I class anti-cancer agent.

8-chlorine adenosine feature is as follows:

1. molecular formula: C ₁₀H ₈N ₅O ₄Cl

2. molecular weight: 301.75

3. structural formula:

4. route of synthesis:

Adenosine NCS 8-Cl-Adenosine

5. anti-tumor activity:

(1) experiment in vitro:

Experiment material: cell strain: human leukemia HL-60, people's gastric cancer MGc80-3

Culture medium: RPMI 1640 culture medium contain the 10-15% calf serum.

Medicine and preparation: 8-chlorine adenosine is by the synthetic (lot number: 930401) by variable concentrations medicine is dissolved in normal saline, high temperature (100 ℃) sterilization or by the mixed fibre resin membrane filtration degerming of 0.22 μ of College of Pharmaceuticals, Beijing Medical University's natural drug and bionical medicine National Key Laboratory.

Experimental technique: cell suspension is pressed finite concentration (HL-60:6 * 10 ⁴/ ml, K562:1.5 * 10 ⁵/ ml, MGc80-3:1 * 10 ⁵/ ml) be inoculated in 24 orifice plates 2.0ml/ hole.Place CO ₂Incubator (37 ℃) press the variable concentrations dosing after cultivating 3h., in the different time counting of drawing materials, calculating inhibitory rate of cell growth and half-inhibition concentration (iC ₅₀).

Experimental result: (table 1-3)

Table 1.8-chlorine adenosine is to the growth inhibited effect of human leukemia cell HL-60

Drug level (μ M) growth inhibition ratio % (48h.)

6.0 68

3.6 63

2.2 58

1.3 48

0.8 32

0.5 18

iC ₅₀＝1.8μM

Table 2.8-chlorine adenosine is to human leukemia cell K ₅₆₂The growth inhibited effect

Drug level (μ M) growth inhibition ratio % (48h)

16.6 65.5

10.0 61.5

6.0 56.1

3.6 48.0

2.2 45.5

1.3 42.2

iC ₅₀＝4.2μM

Table 3.8-chlorine adenosine is to the growth inhibited effect of gastric carcinoma cells MGc80-3

Drug level (μ M) growth inhibition ratio % (48h)

8.0 82

4.0 64

2.0 55

1.0 41

0.5 32

0.25 12

iC ₅₀＝1.56μM

(2) experiment in the body:

Experiment material:

Tumor strain: mouse leukemia L ₁₂₁₀With rat liver cancer H ₂₂

Animal: DBA/2 mouse inbred lines (L ₁₂₁₀The host) and Kunming kind white mice (H ₂₂The host), body weight 18-22g, ♀, ♂ half and half.

Medicine and preparation: 8-chlorine adenosine (the same, lot number: 930721), medicine is dissolved in the normal saline high temperature (100 ℃) sterilization by variable concentrations.

Experimental technique:

Tumor strain inoculation: with L ₁₂₁₀Cell (1.5 * 10 ⁴/ 0.2ml/ is only) be inoculated in the DBA/2 mouse peritoneal; H ₂₂Cell (3 * 10 ⁶/ 0.2ml/ is only) to be inoculated in the Kunming right front axillary fossa of kind white mice subcutaneous.Inoculation back 24h weighs and random packet, presses variable concentrations administration (i.p. or i.v.), and 1 time/day, totally 7 times, H ₂₂Mice with tumor is peeled off tumor in the last administration next day, claims tumor heavy, calculates tumour inhibiting rate; L ₁₂₁₀Mice with tumor is observed survival time of mice (ending to 60 days) and is calculated increase in life span.

Experimental result: (table 4-5)

Table 4. 8-chlorine adenosine is to rat liver cancer solid tumor H ₂₂Curative effect

Heavy (g) suppression ratio % of dosed administration approach number of animals the weight of animals (g) tumor P value

(mg/kg/ day) and number of times begin to finish to begin to finish the treatment contrast

100 i.p.×7 10 10 21.0 28.7 0.68 1.56 56.4 <0.05

i.v.×7 8 8 19.6 22.7 0.42 1.08 61.1 <0.01

60 i.p.×7 10 10 20.1 28.7 0.81 1.56 48.1 <0.01

i.v.×7 8 8 19.4 24.3 0.73 1.08 32.4 <0.05

36 i.p.×7 10 10 21.0 29.5 1.03 1.56 34.0 <0.01

i.v.×7 8 8 20.2 25.5 0.82 1.08 24.1 >0.05

EC ₅₀=63.6mg/kg(i.p.) and 70.8mg/kg(i.v.)

Table 5.8-chlorine adenosine is to mouse leukemia L ₁₂₁₀Preface imitate

Dosed administration approach number of animals average life rate elongation P value

(mg/kg/ days) and number of times existence natural law

100 i.p.×7 6 41.2 122.7 <0.01

i.v.×7 6 40.0 119.8 <0.01

60 i.p.×7 6 41.0 121.6 <0.01

i.v.×7 6 23.8 30.8 <0.01

36 i.p.×7 6 38.0 105.4 <0.01

i.v.×7 6 21.5 18.1 >0.05

6. 8-chlorine adenosine is to the acute toxicity testing of mice

Experiment material:

Animal: Kunming kind white mice, body weight 18-22g, 10 every group (♂, ♀ half and half).

Medicine and preparation: experiment in the consubstantiality.

Experimental technique: with variable concentrations medicine (establishing 10 dosage groups altogether) once abdominal cavity injection, observe the animal acute reaction and write down the death time and the dead animal number, observed altogether 7 days, calculate mortality rate and half lethal dose (LD ₅₀).

Experimental result: (table 6)

Table 6. 8-chlorine adenosine is to acute toxicity and the LD of mice ₅₀

Dosage (mg/kg) number of animals (only) mortality rate %

2000 10 100

1800 10 100

1620 10 100

1458 10 95

1312 10 80

1181 10 65

1063 10 60

957 10 45

861 10 20

775 10 10

LD ₅₀＝1025mg/kg

The LD of table 7. 8-chlorine adenosine and known effective antitumor chemotherapeutic ₅₀Relatively

Medicine LD ₅₀(mg/kg) source of data

1025.0 data of 8-chlorine adenosine

MTX 13.3 Chinese Academy of Sciences

5-Fu 92.6 Shanghai medicines grind

CYT 540.0 studies carefully institute

TSPA 23.3 "

A-130 15.0 "

PCB 430.0 "

BUS 27.1 "

Carmustine 31.6 "

COLC 2.02 "

VLB 3.6 "

VCR 2.21 "

CPT 59.0 "

DACT 0.703 "

MC 10.38 "

7. 8-chlorine adenosine and 8-Cl-cAMP anti-tumor activity are relatively

Experiment material: cell: human leukemia HL-60

Medicine: 8-chlorine adenosine and 8-Cl-cAMP are the synthetic (lot number: 930401) of College of Pharmaceuticals, Beijing Medical University's natural drug and bionical medicine National Key Laboratory.

Culture medium: RPMI1640 contains 15% calf serum.

Experimental technique: add the 8-chlorine adenosine and the 8-Cl-cAMP of same concentrations after the cell routine inoculation respectively, draw materials day by day and calculate cell number and inhibitory rate of cell growth.

Experimental result:

Drug cell growth inhibition ratio %

8-chlorine adenosine (10 μ M) 76.

8-Cl-cAMP(10μM) 57.5

The result shows that 8-chlorine adenosine is bigger by 33.4% to leukemia HL-60 cell inhibitory rate than the 8-Cl-cAMP of same concentrations.

From above pharmacodynamics and toxicological experiment presentation of results, 8-chlorine adenosine has better curative effect to kinds of tumors, and its antitumor spectra is wide, and anti-tumor activity is higher than 8-ClcAMP, and toxicity is far below much being used for clinical chemotherapeutic.

The invention process method: 8-chlorine adenosine can combine with corresponding pharmaceutical carrier and be prepared into various dosage forms and be used for oncotherapy.Embodiment is sub as following table:

Dosage form specification adjuvant route of administration remarks

Injection 250mg water for injection or intramuscular injection comprise that injectable powder reaches

-1000mg other solvent intravenous injection or instillation, the injection of solution agent

And local injection

Tablet 100mg, medical starch or the coated tablet that comprises for oral administration,

250mg, other adjuvant Film coated tablets etc.

500mg.

Capsule 100mg, the capsule for medicine conventional capsule that comprises for oral administration

250mg, and microencapsulation

500mg.

Syrup and 50mg/ml syrup, pure water is for oral administration to comprise that other are oral

Solution liquid

Suppository 100mg, fatty, water-soluble anus comprises vaginal suppository

250mg, property and hydrophilic

500mg/ substrate

3g

Claims (2)

1, prepare antitumor drug with the following 8-chlorine adenosine of structural formula:

2, according to the method for claim 1., wherein said pharmaceutical dosage form is:

Injection (comprising injectable powder and injection of solution agent), tablet (comprising coated tablet, Film coated tablets etc.), capsule (comprising conventional capsule and microencapsulation), syrup and other oral liquids, suppository various dosage forms such as (comprising rectal suppository and vaginal suppository).