JPS63258818A - Carcinostatic agent - Google Patents

Abstract

PURPOSE:To obtain a carcinostatic agent comprising 2'-deoxy-2'-methylidene cytidine or an acid addition salt thereof as an active ingredient. CONSTITUTION:A carcinostatic agent containing a compound shown by the formula or an acid addition salt thereof (e.g. inorganic salt such as hydrochloride or hydrobromide and organic acid salt such as maleate) as an active ingredient. Powder, granule, tablet, sugar coated tablet, capsule, syrup, suppository, external preparation, injection or agent for drip may be cited as the dosage form. Oral administration is especially preferable. A dose is 10-400mg/kg/day, preferably 50-200mg/kg/day in the case of oral agent and 1-10mg/kg/day, preferably 1-5mg/kg/day in the case of injection. The number of times of administration is once-four times daily and can be properly selected. The carcinostatic agent can be used with another carcinostatic agent, immunoactivator or other drugs.

Description

[Detailed description of the invention] [Industrial application field] TECHNICAL FIELD The present invention relates to medicines, particularly anticancer agents.

[Conventional technology]

With cancer deaths increasing, various types of chemotherapy and immunotherapy are being used in conjunction with surgical therapy. In chemotherapy, cytosin arabinoside, 5-fluorouracil, and the like are clinically used as antimetabolite antineoplastic agents, as they are effective for acute leukemia and the like.

[Problems to be solved by the invention]

However, conventional anticancer drugs still have various problems, such as not providing a fully satisfactory healing effect and causing side effects, and there is a demand for the development of better anticancer drugs from various fields.

[Means for solving problems]

Therefore, from this point of view, the present inventors have conducted many years of research, and as a result, a new nucleondo compound has been discovered.
The present invention was completed based on the discovery that it has an antitumor effect on tumor cells such as leukemia or P388 leukemia.

The present invention relates to an anticancer agent containing 2°-deoxy-2°-methylidenecytidine or a pharmaceutically acceptable acid addition salt thereof as an active ingredient.

2°-deoxy-2′-methylidenecytidine is HO
The pharmaceutically acceptable acid addition salts thereof include inorganic acid salts such as hydrochloride, hydrobromide, sulfate, phosphate, maleate, fumarate, tartrate, Examples include organic acid salts such as succinate, citrate, p-)luenesulfonate, and pamoate.

A typical active ingredient compound of the present invention is 2°-deoxy-2°-methylidenecytidine hydrochloride 1/2 hydrate [
Melting point: 148-155°C (decomposition point). Hereinafter, it is also referred to as compound A].

[For production]

Next, the antitumor effect of the active ingredient compound of the present invention will be explained using pharmacological experiments.

Experimental Example 1 After dissolving Compound A in dimethyl sulfoxide, serial dilution was performed with distilled water for injection to a final concentration of 20 times.
1.0.0.5.0.125 and 0.0625μg/
The test solution had a concentration of ml. Add 10 μl of this test solution to 96
Place in a hole tissue culture plate (Fashion Na3072), add 190 μl of L1210 leukemia cell suspension containing I x 10 cells/ml, and incubate in a carbon dioxide gas incubator for 3
The cells were cultured at 7°C for 48 hours. Distilled water for injection as a control group
A similar culture using only Oμe was used.

After culturing, the number of living cells was counted using an erythrocyte counter using the trypan blue staining method, and the Ml tumor growth rate was determined.
50% tumor growth inhibitory concentration of Compound A (ICs μg/m
l) was 0.12 μg/ml.

The final concentration of dimethyl sulfoxide is 0.0.
In a control group diluted in the same manner with dimethyl sulfoxide alone at 1% or less, no effect on tumor cell growth was observed.

Experimental Example 2 Group 3 female CD F + mice (8 weeks old) were given 106
P388 leukemia (derived from the US National Cancer Institute) was intraperitoneally transplanted, and Compound A was continuously administered intraperitoneally (i, p,) once a day for 5 days starting from the day after the transplant. Median survival days (me
Based on this, the life extension rate (T/C, %) is calculated using the following formula.
The results are summarized in Table 1.

MST of control group Table 1 100 14.0 149 As is clear from the results shown in Table 1, the active ingredient compound of the present invention has no effect on T.
The /C value was 125% or more, indicating that the drug was effective and that the effect was dose-dependent.

The above experimental results show that the active ingredient compound of the present invention is useful as an anticancer agent for various mammals including humans. It also has low toxicity and can be safely administered.

The active ingredient compound of the present invention is used as an anticancer agent. It can be mixed with swelling agents, diluents, etc. to form powders, granules, tablets, sugar-coated tablets, capsules, syrups, physiological preparations, external preparations, injections, and infusion preparations. Possible.

The anticancer agent of the present invention is administered orally or parenterally to mammals including humans, with oral administration being particularly preferred. The dose may vary as long as it effectively inhibits the target cancer, and may vary depending on the animal to be treated, the type of cancer, the route of administration, the dosage form, etc., but generally in the case of oral preparations, the dose is 10 to 100 liters per day.
400 mg/kg body weight, preferably 50-200 akebono/k
g body weight, and for injections it is 1 to 10 ■/kg body weight per day,
Preferably it is 1 to 5 .mu./kg body weight. The frequency of administration can be appropriately selected within the range of 1 to 4 times a day.

The anticancer agent of the present invention can be used in combination with other anticancer agents, immunostimulants, or other acceptable drugs.

〔Example〕

The present invention will be explained below by showing production examples of Compound A and formulation examples of the drug of the present invention, but the present invention is not limited by these in any way.

Production example 2'-deoxy-2'-methylidenecytidine hydrochloride production +11 1-β-D-ribofuranosyl-4-ethoxy-
Synthesis of 2-pyrimidone 2',3',5°-tri0-acetyluricif3.
Dissolve 35g in 50IIll of chloroform, add 8.1ml of thionyl chloride and 0.5ml of dimethylformamide.
After refluxing for 6 hours and 30 minutes, the mixture was dried under slight pressure. The residual aroma was dissolved in 20 Ill of ethanol, 30 ml of 1N sodium ethoxide was added, and after refluxing for 2 hours, it was neutralized with 1N hydrochloric acid, the precipitated salt was filtered off, and the solution was concentrated to dryness. Add this to a silica gel column (4X31c+
a), and the target compound-containing fraction was extracted with 16% ethanol-chloroform, and the solvent was distilled off to obtain crude crystals of the target compound.

This was recrystallized from ethanol to obtain 2.08 g (yield: 84.2%) of the desired product (as a 1/3 hydrate).

Melting point: 136-137.5°C Elemental analysis value calculated as CzH+JzOs·1/3 ozo C: 46.97%, H: 6.09%.

N: 9.96%, 0: 36.98% actual value C:
46.91%, H: 6.02%.

N: 9.98%, O17,09%f211-(2
-oxo-3,5-0-tetraisopropyldisiloxanyl-β-D-ribofuranosyl)-4-ethoxy-2-
Synthesis of pyrimidone Dissolve 7.04 g of 1-β-D-ribofuranosyl-4-ethoxy-2-pyrrolidone in 80 ml of pyridine, cool with water, and then prepare 1,3-dichloro-1,1,3°3-tetraisopropyldisiloxane. 9.57 g was added, and the mixture was stirred and reacted at room temperature for 4 hours and 30 minutes. Ice water was added, the solvent was distilled off, the residual aroma was distributed between chloroform and water, the chloroform layer was dried, the solvent was distilled off, and the residual aroma was transferred to a silica gel column (1
The eluted portion was collected and concentrated with 40% ethyl acetate-hexane.
12.3 g of 0-tetraisopropyldisiloxane was obtained.

Next, add 2.7 ml of oxalyl chloride to 40 ml of methylene chloride.
1 and cooled to -70°C. Under an argon stream, 4.81 parts of dimethyl sulfoxide dissolved in 20 parts of methylene chloride was added dropwise over 20 minutes, followed by stirring for 30 minutes. To this, 12.3 g of the above 3',5'-〇-tetraisopropyl disiloxane dissolved in 50 ml of methylene chloride was added dropwise, and after stirring at -70°C for 2 hours, 20 ml of triethylamine was added and the mixture was further stirred for 1 hour. did. This reaction solution was returned to room temperature, water was added and distributed,
The methylene chloride layer was separated, the solvent was distilled off, the residual aroma was dissolved in ethyl acetate, and the solution was partitioned with water. The ethyl acetate layer was concentrated to dryness, adsorbed on a silica gel column (5X2BC11), and both fractions containing the target substance eluted with 20% ethyl acetate-hexane were collected. After the solvent was distilled off, the target substance 10 was crystallized from hexane. .2 g (yield 72.1%) was obtained.

Melting point: 157.5-159°C Elemental analysis value: Calculated value as CtslhJzOtSi2 C: 53.87%, H near, 86%.

N: 5.46% Actual value C: 53.73%, H near, 87%.

N: 5.51% (311-(2-deoxy-2-methylidene-β-D-
Synthesis of (ribofuranosyl)-4-ethoxy-2-pyrimidone 232N of potassium hydride was added to 2.4 ml of dimethyl sulfoxide under an argon stream, and the mixture was stirred at room temperature for 40 minutes.

2.2 g of methyltriphenylphosphonium bromide was dissolved in 8 ml of dimethyl sulfoxide, and the above mixture of potassium hydride and dimethyl sulfoxide was added dropwise thereto under water cooling in an argon stream, followed by stirring for 10 minutes. Add to this 1 above
1.02 g of crystals of -(2-oxo-3,5-0-tetraisopropyldisiloxanyl-β-D-ribofuranosyl)-4-ethoxy-2-pyrimidone was dissolved in 10 ml of dimethyl sulfoxide and the solution was heated under an argon atmosphere. The mixture was added dropwise at the bottom and stirred for 2 hours under water cooling. Add 10ml of 1N ammonium chloride aqueous solution to this, and add 50ml of ethyl acetate.
1 and 40 ml of water were added and distributed. The organic layer was concentrated under reduced pressure and adsorbed on a silica gel column (2,4X30c+a),
Elution was performed with a hexane-ethyl acetate mixed solvent to obtain a 2°-methylidened compound.

320 ■ of the compound obtained above was added to 1 of tetrahydrofuran.
0+++1, add 1 ml of tetra n-butylammonium fluoride, and leave in a room. Stirred for mlO minutes.

After neutralization with acetic acid, silica gel column (2,4X12(J)
The product was adsorbed onto the solution, eluted with chloroform-ethanol, and the eluted fractions containing the target compound were collected to obtain 155 ml of deprotected 2°-methylidene-4-〇-ethyl compound (yield: 30%).

Melting point: 157.5-159°C Elemental analysis value: Calculated value as C+J+JhOs C:5
3.12%, H: 6.01%.

N: 10.44% Actual value C: 53.80%, I (j 5.99%.

N: 10.37% (4) Synthesis of 2°-deoxy-2°-methylidenecytidine hydrochloride/half hydrate 150 2°-methylidene-4-〇-ethyl compound was added to ammonia-saturated methanol under water cooling. Dissolve in 10ml of solution,
It was placed in a sealed tube and heated at 100''c for 2 days.

After cooling, 2 ml of 2N hydrochloric acid was added, concentrated, and crystallized from ethanol-water to give the title compound, 125 ml (yield: 81.
7%).

Melting point: 148-155°C (decomposition point) Elemental analysis value: C+all+JiOn ・HCI
- Calculated values as 1/2 HzO: C: 42.14%, H: 5.31%.

N: 14.74% Actual values: C: 42.25%, H: 5.26%.

N: 14.69% Formulation Example 1 Compound A 50.0 ■ Finely powdered cellulose 25.0 ■ Milk $1!
49. s■ Starch 40. Owt Talc 5. Tablet consisting of θ■magnesium stearate 0.5■.

Sugar-coated tablets or film-coated tablets can be obtained by sugar-coating or film-coating, if desired.

Formulation Example 2 Compound A 50.0 ■Milk*
S O, O ■ Starch 15.0 ■ Talc 5. Capsules made by filling capsules with the composition θ■.

Formulation example 3 Compound A 10% milk $7
! Fine granules consisting of 80% starch and 10%.

Formulation Example 4 Compound A 10% Milk W
Granules consisting of 55% finely powdered cellulose 20% starch 15%.

Formulation Example 5 Compound A 50.0 ■Glucose 100.0 Akebono is dissolved in purified water to prepare an injection solution with a total volume of 2s+1.

Formulation Example 6 Compound A 1100n Witepsol ■ H2S 950 Akebono Witepsol (l H7S 950g) However, Witepsol is manufactured by Wuitten (West Germany)
It is a registered trademark owned by the company.

Formulation Example 7 Compound A 2g Ethyl parahydroxybenzoate 0.025g Propyl parahydroxybenzoate 0.015g Sodium lauryl sulfate 1.5g Propylene glycol
12. Og stearyl alcohol
22.0g white petrolatum
Dissolve 25.0 g of the composition in purified water to make a total amount of 10
0.0g of hydrophilic ointment.

Claims (1)

[Claims] An anticancer agent containing 2'-deoxy-2'-methylidenecytidine or a pharmaceutically acceptable acid addition salt thereof as an active ingredient.