JPH02225471A - Imidazole derivative and antitumor agent - Google Patents

Abstract

NEW MATERIAL:Imidazole derivatives expressed by formula I (A is CONH2 or CN; R<1> is H, lower alkyl, hydroxy-lower alkyl or phenyl) or salts thereof. EXAMPLE:5-Ethynylimidazole-4-carbonitrile. USE:An antitumor agent, having excellent antitumor action and a simple structure and capable of being readily synthesized. PREPARATION:A compound expressed by formula II (X is halogen) is reacted with a compound expressed by the formula HCidenticalCR<1>' (R<1>' is R<1> except H) to afford a compound expressed by formula III in the compounds expressed by formula I. Alternatively, the compound expressed by formula II is reacted with a compound expressed by the formula CHidenticalCY (Y is silyl) to introduce silylethynyl group into the 5-position of the imidazole ring. The silyl group is then removed to afford the exemplified compound expressed by formula I (A is CN; R<1> is H).

Description

Detailed Description of the Invention [Industrial Application Field] The present invention relates to an imidazole derivative and an antitumor agent containing it as an active ingredient. -Alkynyl-1-β-D-ribofuranosylimidazole-4-carboxamide was found to have excellent antitumor activity (Chem, Pha
rm, Bull, 36 (7), 2730 (1
988)), and 5-alkynyl-1-β-D-ribofuranosylimidazole-4-carbonitrile was also found to have antitumor effects (Japanese Patent Application No. 63-82).
No. 089).

[Problem to be solved by the invention]

The above-mentioned 5-alkynylimidazole nucleosides are:
Because they are nucleosides, they are not easy to synthesize, and some of them lack stability.

[Means to solve the problem]

The present inventors have conducted various studies on compounds that have excellent antitumor effects, have a simple structure, and can be easily synthesized. As a result, we succeeded in synthesizing imidazole derivatives represented by the following formula [1], and also found that these imidazole derivatives have excellent antitumor effects.

That is, the present invention provides the following formula: This relates to an imidazole derivative or a salt thereof represented by the following.

The present invention also relates to an antitumor agent containing the compound of the present invention represented by formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient.

Hereinafter, the compounds of the present invention will be explained in detail.

The compound of the present invention is represented by the above formula (1). - In formula CI), the lower alkyl group represented by R1 is a linear or branched alkyl group having any number of carbon atoms, preferably 10 or less, more preferably 4 or less. Can be mentioned. Furthermore, in view of the antitumor activity described below, preferable examples include straight-chain or branched alkyl groups having 1 to 3 carbon atoms, such as methyl, ethyl, propyl, and isopropyl. Further, the hydroxy lower alkyl group represented by R1 is preferably a linear or branched alkyl group having any number of carbon atoms of 10 or less, and any number of hydroxyl groups substituted at any position. and hydroxy lower alkyl groups. Furthermore, like the above-mentioned alkyl groups, hydroxymethyl, α-hydroxyethyl, β-hydroxyethyl, α,β-dihydroxyethyl, α-hydroxypropyl, β-hydroxypropyl,
γ-hydroxypropyl, α,β-dihydroxypropyl, α,γ-dihydroxypropyl, β,γ-dihydroxypropyl, α-hydroxymethylethyl, β-hydroxymethylethyl, α,β-dihydroxymethylethyl, α-hydroxybutyl , β-hydroxybutyl, γ
-Hydroxybutyl, δ-hydroxybutyl, α, β-
Dihydroxybutyl, α, γ-dihydroxybutyl, α
, δ-dihydroxybutyl, α, β, γ-trihydroxybutyl, β, γ, δ-trihydroxybutyl, and other hydroxy lower alkyl groups having 1 to 4 carbon atoms substituted with a hydroxyl group at any position. can be exemplified.

Specific examples of compounds of the present invention in which A is a carbamoyl group in General 2H) include 5-(phenylethyne-1-
yl) imidazole-4-carboxamide, 5-ethynylimidazole-4-carboxamide, 5-(1-propyn-1-yl)imidazole-4-carboxamide,
5-(1-butyn-?1-yl)imidazole-4-carboxamide, 5-(1-pentyn-1-yl)imidazole-4-carboxamide, 5-(1-hexyne-
1-yl)imidazole-4-carboxamide, 5-(
1-heptyn-1-yl)imidazole-4-carboxamide, 5-(1-octyn-1-yl)imidazole-4-carboxamide, 5-(1-nonyn-1-yl)
Imidazole-4-carboxamide, 5-(1-decyn-1-yl)imidazole-4-carboxamide, 5-
(1-undecin-1-yl)imidazole-4-carboxamide, 5-(1-dobcin-1-yl)imidazole-4-carboxamide, 5-(3-methyl-1-yl)
butyn-1-yl)imidazole-4-carboxamide, 5-(3,3-dimethyl-1-propyn-1-yl)imidazole-4-carboxamide, 5-(3-methyl-1-pentyn-1-yl) Imidazole-4-carboxamide.

5-(4-methyl-1-pentyn-1-yl)imidazole-4-carboxamide, 5-(3-hydroxy-
1-propyn-1-yl)imidazole-4-carboxamide, 5-(3-hydroxy-1-butyn-1-yl)imidazole-4-carboxamide, 5-(4-hydroxy-1-butyn-1-yl) Imidazole-4-carboxamide, 5-(3,4-dihydroxy-1-butyn-1-yl)imidazole-4-carboxamide,
5-(3-hydroxy-1-pentyn-1-yl)imidazole-4-carboxamide, 5-(4-hydroxy-1-pentyn-1-yl)imidazole-4-carboxamide, 5-(5-hydroxy-1 -Pentyne-1-
yl) imidazole-4-carpoxamide, 5-(3
, 4-dihydroxy-1-pentyn-1-yl)imidazole-4-carboxamide, 5-(4,5-dihydroxy-1-pentyn-1-yl)imidazole-4-carboxamide, 5-(4-hydroxy-3 -methyl-1
-butyn-1-yl)imidazole-4-carpoxamide, 5-(6-hydroxy-1-hexyn-1-yl)
Imidazole-4-carboxamide, 5-(4-hydroxy-1-hexyn-1-yl)imidazole-4-carboxamide.

5-(3-methyl-5-hydroxy-1-pentyne-1
Examples include compounds such as -yl)imidazole-4-carboxamide.

- Specific examples of the compounds of the present invention in which A is a cyano group in formula [1] include 5-(phenylethyn-1-yl)imidazole-4-carbonitrile, 5-ethynylimidazole-4-carbonitrile, 5-( 1-propyn-1-yl)imidazole-4-carbonitrile, 5-
(1-Butyn-1-yl)imidazole-4-carbonitrile.

5-(1-pentyn-1-yl)imidazole-4-carbonitrile, 5-(1-hexyn-1-yl)imidazole-4-carbonitrile, 5-(1-heptyn-
1-yl)imidazole-4-carponitrile, 5-(
1-Octyn-1-yl)imidazole-4-carbonitrile, 5-(1-nonyn-1-yl)imidazole-4-carbonitrile, 5-(1-decyn-1-yl)
Imidazole-4-carbonitrile, 5-(1-undecyn-1-yl)imidazole-4-carbonitrile,
5-(1-dobcin-1-yl)imidazole-4-carbonitrile, 5-(3-methyl-1-butyn-1-
imidazole-4-carbonitrile, 5-(3
, 3-dimethyl-1-propyn-1-yl)imidazole-4-carbonitrile, 5-(3-methyl-1-pentyn-1-yl)imidazole-4-carbonitrile,
5-(4-methyl-1-pentyn-1-yl)imidazole-4-carbonitrile, 5-(3-hydroxy-1
-propyn-1-yl)imidazole-4-carbonitrile, 5-(3-hydroxy-1-butyn-1-yl)
Imidazole-4-carbonitrile, 5-(4-hydroxy-1-butyn-1-yl)imidazole-4-carbonitrile, 5-(3,4-dihydroxy-1-butyn-1-yl)imidazole-4- carbonitrile, 5
- (3-hydroxy-1-pentyn-1-yl)imidazole-4-carbonitrile, 5- (4-hydroxy-1-pentyn-1-yl)imidazole-4-carbonitrile, 5-(5-hydroxy- 1-pentyne-1
-yl)imidazole-4-carbonitrile, 5-(
3,4-dihydroxy-1-pentyn-1-yl)imidazole-4-carbonitrile, 5-(4,5-dihydroxy-1-pentyn-1-yl)imidazole-4
-Carbonitrile, 5-(4-hydroxy-3-methyl-1-butyn-1-yl)imidazole-4-carbonitrile, 5-(6-hydroxy-1-hexyne-1-
yl) imidazole-4-carbonitrile.

5-(4-hydroxy-1-hexyn-1-yl)imidazole-4-carbonitrile, 5-(3-methyl-
Examples include compounds such as 5-hydroxy-1-pentyn-1-yl)imidazole-4-carbonitrile.

The salt of the compound represented by formula (1) may be any salt of the compound with an ion having a charge capable of forming a salt, and pharmaceutically acceptable salts are particularly preferred. Examples of salts include salts of the compound of general formula (f) with cations such as sodium, potassium, and calcium, or inorganic acids (hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, etc.) or organic acids (p-toluenesulfonic acid, etc.). ) and other anion salts.

Hereinafter, the method for synthesizing the compound of the present invention will be explained in detail.

The compound of the present invention represented by general formula (1) can be easily synthesized using known compounds as raw materials.

- The compound of the present invention represented by the formula [■'] [wherein A has the same meaning as defined above, and R'' represents a lower alkyl group, a hydroxy lower alkyl group, or a phenyl group].

- Formula (n) Above-1 In the formula C, A has the same meaning as above or represents a halogen atom, and - a 5-halogenoimidazole derivative represented by the formula [■] HCECR'' Cm) [in the formula ,
R'' has the same meaning as above.] (Hereinafter, the above method will be referred to as Synthesis Method I).

In the above-mentioned formula [■], examples of the halogen atom represented by X include iodine, bromine, fluorine, and chlorine.

In the compounds represented by the general formula [■] having a halogen atom, all 5-halogenoimidazole-4-carboxamides in which A is a carbamoyl group are known compounds. 5-iodoimidazole-4-carboxamide (CA (36062-66-5)), 5-bromoimidazole-4-carboxamide (CA, [36
062-67-63) and 5-chloroimidazole-
4-Carboxamide (CA [36062-68-7]
) for Cham, Phrm, Bull, , upper 9
(8).

1737 (1971). 5-Fluoroimidazole-4-carboxamide (CA(3330)
0-35-5)) for Life Sci.

↓7 (2), 187 (1975).

Furthermore, in the compound represented by formula [■], 5-halogenoimidazole-4-carbonitrile in which A is a cyano group is a known compound, 5-amizimidazole-4-carbonitrile (CA (5098-11 =33
(see JP-A-49-116063)) can be synthesized by halogenating the 5-position of the imidazole ring.

The above halogenation reaction is a halogenation reaction via a diazonium compound, for example, Na1r and Rich
Ardson's method (J, Org, Cham, 45
, 3969-3974. (1980)).

Specifically, as the halogenating agent, tribromomethane can be used for bromination, and short methane can be used for iodination, and these halogenating agents also act as reaction solvents. The halogenation reaction is carried out by dissolving the above raw material compound and alkyl nitrite (for example, isoamyl nitrite, butyl nitrite, etc.) in a halogenating agent in an amount of 2 to 30 times the mole of the compound, and refluxing the solvent at 50°C. It can be carried out by reacting at a temperature for 10 minutes to 30 hours.

The acetylene derivative represented by the above-mentioned formula [■] may be selected from those having R1' corresponding to the desired type of compound represented by the general formula [2].

When R'' is a lower alkyl group or a hydroxy lower alkyl group, specific examples thereof include those exemplified for R1.

The reaction for introducing an alkynyl group in Synthesis Method I is carried out in a reaction solvent, preferably in the presence of a palladium catalyst, -1 to 3 times the mole of the compound of general formula (III) per mole of the compound of formula [1], preferably 1 ~1 reaction temperature using ~2 times the mole ~50 ~
It can be carried out by reacting at 150°C, preferably 90 to 110°C, for 1 to 30 hours.

As a reaction solvent, basic solvents (triethylamine, tributylamine, trioctylamine, N, N, N'
, N'-tetramethyl-1,8-naphthalenediamine,
dimethylaniline, diethylaniline, pyridine, etc.)
Alone or with these basic solvents, acetonitrile, N,
N-dimethylformamide, dimethyl sulfoxide, N
A mixed solvent with a solvent such as , N-dimethylacetamide, tetrahydrofuran, or dioxane can be used.

Examples of the palladium catalyst include bis(acetonitrile)palladium dichloride, bis(triphenylphosphine)palladium dichloride, bis(benzonitrile)palladium dichloride, and tetrakis(triphenylphosphine)palladium.

Isolation and purification of the compound of general formula [1'] from the reaction solution should be performed by methods commonly used for the isolation and purification of nucleic acid bases (e.g., various chromatography methods such as adsorption or ion exchange, recrystallization methods, etc.) Can be done.

A compound in which A is a cyano group and R1 is a hydrogen atom in the general formula (1), that is, 5-ethynylimidazole-4
-Carbonitrile is a compound represented by the above-mentioned formula [n] and -formula [■']HC20Y (II
I') Synthesis by reacting a silylated acetylene derivative represented by [wherein Y represents a silyl group] to introduce a silylated ethynyl group into the 5-position of the imidazole ring, and then removing the silyl group. (Hereinafter, the above method will be referred to as synthesis method ■).

In the above formula [■'], the silyl group represented by Y is a silyl group that is commonly used in the synthesis of organic compounds such as trimethylsilyl, t-butyldimethylsilyl, methyldi-t-butylsilyl, tripropylsilyl, etc. I can give an example.

- The reaction of reacting the compound of formula (If) with the compound of general formula [■'] to introduce a silylated ethynyl group at the 5-position of the imidazole ring is carried out in the same reaction solvent as in Synthesis Method I, in the presence of the same catalyst, It can be carried out using similar reaction conditions.

Furthermore, the reaction for removing the silyl group may be carried out according to conventional methods, such as acidic hydrolysis using hydrochloric acid-tetrahydrofuran-water, alkaline hydrolysis using methanol/ammonia, treatment with ammonium fluoride, tetrabutylammonium fluoride, etc. It can be removed by etc.

The target compound can be isolated and purified from the reaction solution by a method commonly used for the isolation and purification of nucleic acid bases.

A compound in which R'' is a hydrogen atom in the general formula [I], that is, 5-ethynyl imidazole-4-carboxamide or 5-ethynyl imidazole-4-carbonitrile, is represented by the above-mentioned formula (II) It can be synthesized by reacting the compound represented by the formula with an ethynyltin compound to introduce a silylated ethynyl group into the 5-position of the imidazole ring, and then removing the silyl group (hereinafter, the above method will be referred to as synthesis method Ⅰ). .

As the ethynyltin compound used in the above reaction, a compound represented by the formula (IVI Z, SnCmic y (IV) [wherein, Y has the same meaning as above, and 2 represents a lower alkyl group] is used. - In the formula (IV), the silyl group represented by Y can be exemplified by those exemplified in the synthesis method ①. Also, as the lower alkyl group for Z, methyl, ethyl, propyl, Examples include alkyl groups having 1 to 4 carbon atoms such as butyl.

Such ethynyltin compounds include 5tills and Si
mpson method (J, Am, Che+m, Soc, .
109, 2138-2152 (1987)).

In the reaction for introducing a silylated ethynyl group into the 5-position of the imidazole ring, the compound represented by the formula [■] and the ethynyltin compound are mixed in one reaction solvent (for example, N,N-dimethylformamide, N.

The reaction can be carried out in a solvent such as N-dimethylacetamide, dimethylsulfoxide, acetonitrile, tetrahydrofuran, dioxane, etc.).

The above reaction proceeds efficiently when carried out in the presence of a palladium catalyst. Examples of such palladium catalysts include the catalysts exemplified as those used in 2 Synthesis Method I.

The above reaction is carried out using an ethynyltin compound in an amount of 1 to 2 times the mole of the compound represented by formula [■], and preferably a catalytic amount of a palladium catalyst, at a reaction temperature of 0.
It can be carried out by reacting at ~200°C, preferably 50-150°C for 1-30 hours.

The compound obtained by the above reaction and having a silylated ethynyl group introduced at the 5-position is isolated and purified, if necessary, and subjected to a reaction to remove the primary silyl group.

The reaction for removing the silyl group can be carried out in the same manner as in Synthesis (2).

After the reaction, the obtained target compound can be isolated and purified by methods commonly used for isolating and purifying nucleic acid bases (eg, various chromatographic recrystallization methods such as adsorption or ion exchange).

A compound in which A is a carbamoyl group in the general formula (1) is obtained by subjecting a corresponding compound in which A is a cyano group in formula (17) to a hydrolysis reaction to form a compound at the 4-position of the imidazole ring of the compound. obtained by converting the cyano group of into a carbamoyl group (hereinafter, the above method is referred to as synthesis method ■)
.

- The raw material compound in which A is a cyano group in formula (1) can be synthesized from known compounds by the synthesis methods ① to ① above.

For the hydrolysis reaction of synthesis method (2), the hydrolysis reaction employed when preparing acid amide derivatives from nitrile derivatives can be applied. Specifically, any one of acidic hydrolysis reaction, neutral hydrolysis reaction, and alkaline hydrolysis reaction, preferably neutral hydrolysis reaction or alkaline hydrolysis reaction, more preferably alkaline hydrolysis reaction applies.

To explain the alkaline hydrolysis reaction more specifically, the raw material compound and hydrogen peroxide are mixed at 10 to 50°C, preferably in the presence of a base such as sodium hydroxide, potassium hydroxide, or ammonium hydroxide (or ammonia). The hydrolysis reaction can be carried out by reacting at room temperature for 1 minute to 10 hours.

In addition, the reaction solvent may be any solvent as long as it dissolves the raw material compound and the 'lR target compound and does not interfere with the above reaction. Specifically, alcoholic solvents such as methanol and ethanol can be used. .

The 5-alkynylimidazole-4-carboxamide thus obtained is isolated and purified by appropriately combining methods commonly used for isolating and purifying nucleic acid bases (for example, various chromatography methods such as adsorption or ion exchange, recrystallization method, etc.). can do.

Synthesis method ■ The compound of the present invention represented by the general formula [I] has the formula [V] [wherein A represents a carbamoyl group or a cyan group, and R
1'' represents a hydrogen atom, a silyl group, a lower alkyl group, a hydroxy lower alkyl group, or a phenyl group, and R2 represents a hydrogen atom, a protecting group for a hydroxyl group, or a phosphoric acid residue (-p○(OH)
), or >P○(OH), etc.) R3 and R
4 are simultaneously OR'', or when one is a hydrogen atom, the other is OR2] It can be synthesized by subjecting an imidazole nucleoside derivative represented by the following to a glycosidic bond cleavage reaction (hereinafter, The above method is called the synthesis method■).

In the above formula (V), when R1' is a lower alkyl group or a hydroxy lower alkyl group, specific examples thereof include the same substituents as those exemplified for aL, and R1'' is a silyl group. In this case, specific examples thereof include the silyl group listed as an example of the silyl group represented by Y in the compound represented by the general formula [I■'] in Synthesis Method H above.

Specific examples of the imidazole nucleoside derivative represented by the general formula [■] include Chem and Pharm.

Examples include known compounds described in J. Bull., 36 (7), 2730 (1988).

The above reaction can basically be carried out by applying the method commonly used for the cleavage reaction of glycosidic bonds of nucleosides with appropriate modifications depending on the type of raw material compound and target compound. A conventional reaction method for a decomposition reaction may be followed.

More specifically, in the presence of an inorganic acid such as hydrochloric acid, sulfuric acid, or hydrobromic acid or an organic acid such as methanesulfonic acid or p-toluenesulfonic acid, in water or a water-containing solvent at a temperature of 50'C to reflux. Several tens of minutes to several tens of hours, preferably 1
This reaction can be carried out by reacting for ~5 hours.

In addition, considering the solubility of the raw material compound and the target compound,
In order to dissolve these compounds, an appropriate solvent such as alcohols such as methanol, ethanol and propatool, and ethers such as dioxane, tetrahydrofuran and diethyl ether may be added to the reaction solution.

In addition, in the above formula (V), when R'' is a silyl group, the silyl group is also removed by subjecting it to the above reaction, and -
A 5-ethynyl imidazole derivative in which R1 is a hydrogen atom in formula [1] is produced.

After the reaction, - the target compound represented by formula (1) is isolated and purified using an appropriate combination of methods commonly used for isolating and purifying nucleic acid bases (for example, various chromatography methods such as adsorption or ion exchange, recrystallization methods, etc.) can do.

The uses of the compound of the present invention will be explained below.

- The imidazole derivative represented by formula (1) has antitumor activity, and the drug of the present invention containing these as an active ingredient is used in tumor treatment.

The dosage of the compound represented by general formula (1), which is the active ingredient of the drug of the present invention, varies depending on the patient's severity of illness, tolerance to the drug, etc., and should be ultimately determined by the physician's judgment. However, it is usually 0.1 to Lo per day for adults.
g is suitable and can be administered once or in divided doses.

The administration method can be any method depending on the administration route.

The medicament of the present invention can be prepared for administration by any conventional method of formulation. Therefore, the drug of the present invention includes a pharmaceutical composition containing as an active ingredient an imidazole derivative represented by general formula (I) suitable for use as a human medicine.

Such compositions are subjected to administration in a conventional manner with any required pharmaceutical carriers or adjuvants.

For example, in the case of a composition preparation for oral administration, it is provided in a form suitable for absorption from the gastrointestinal tract, such as solid preparations such as tablets, capsules, powders, sugar-coated tablets, and granules, syrups,
In the case of solid preparations, syrup, gum arabic, gelatin,
Sorvit.

Binders such as tragacanth, polyvinylpyrrolidone, dextrin, excipients such as lactose, sugar, corn starch, calcium phosphate, sorbitol, glycine, lubricants such as magnesium stearate, talc, polyethylene glycol, silica, disintegration such as potato starch Adjuvants such as agents, wetting agents, stabilizers, and flavoring agents can be selected and used in formulations based on pharmaceutical considerations. For liquid formulations, sorbitol, syrup, methylcellulose, glucose/sugar syrup, gelatin,
Suspending agents such as hydroxyethyl cellulose, carboxymethyl cellulose, aluminum stearate gel, hydrogenated edible oils, emulsifying agents, preservatives such as methyl p-hydroxybenzoate, propyl p-hydroxybenzoate, sorbic acid, etc. can be used.

In addition, when preparing a composition formulation for injection administration, pH adjusters, buffers, stabilizers, preservatives, solubilizers, etc. are added to the compound of the present invention, which is the active ingredient of the present invention, as necessary. Inject it subcutaneously, intramuscularly, intravenously, etc. using conventional methods.

〔Effect of the invention〕

Hereinafter, a method for testing antitumor activity and results of measuring antitumor activity of the compound of the present invention represented by general formula (1) using the method will be explained.

In vitro cell proliferation inhibitory activity against cells derived from mouse tumor (test method 1) ■ Mouse leukemia cells LL210 were cultured in R, PM11640 medium (added with 10% tallow serum), and in the logarithmic growth phase
．． A diluted cell suspension is obtained by diluting with the above medium to 6×10′ cells/−.

■ Add 0.1 d of the test solution serially diluted with culture medium or phosphate buffered saline (PBS) to a test tube containing 0.9 d of the above cell core temperature dilution solution, and incubate at 37°C in a 2% carbon dioxide incubator. , and culture for 48 hours.

(2) After completion of the culture, count the number of cells in each tube using a Coulter Counter (manufactured by Coulter Electronics), and determine the inhibition rate (%) of cell growth at each concentration of the test compound using the formula below.

Cx-C.

Tx: Number of cells at the end of culture in a test tube containing the test compound (cells/-) Cx: Number of cells at the end of culture in a test tube not containing the test compound (
cells/-) Co: Number of cells at the start of culture in test tubes containing no test compound (
/-) ② From the graph of growth inhibition rate and test compound concentration, determine the test compound concentration corresponding to 50% growth inhibition rate and define it as the 50% cell growth inhibition concentration (IC, .).

Test results ■IC, similar to test method 1. seek.

Test results (test method 2) ■ Mouse lymph lung-derived cells P388 or mouse sarcoma-derived cells Sarcoma 180 were suspended in PRM11640 medium (added with 10% tallow serum and 2 μM 2-hydroxyethyl disulfide) at a density of 10 cells/mQ. do. Test compounds are appropriately serially diluted in the same medium.

■ Cell suspension 0.5- and medium or test compound solution 0.5-.
5mQ and cultured in a 5% carbon dioxide incubator at 37°C for 48 hours. The culture is performed in a glass culture test tube.

(2) After completion of the culture, count the number of cells in each culture test tube using a Coulter counter, and determine the growth inhibition rate (%) using the same formula as in Test Method 1.

〔Example〕

Hereinafter, the present invention will be specifically explained with reference to Reference Examples and Examples.

Reference Example 1 Synthesis of 5-iodo-1-(2,3,5-tri〇-acetyl-β-D-ribofuranosyl)imidazole-4-carbonitrile 5-amino-1-(2,3,5-tri〇- Acetyl-β-D-ribofuranosyl)imidazole-4-carbonitrile 10.88g (29.7mm01) was heated at 100°C.
After the reaction, the reaction solution was dissolved in 150 mA of short methane in an oil bath, and 15-isoamyl nitrite was added and reacted for 30 minutes.
The fraction containing the target compound was obtained by elution with 1-4% ethanol-chloroform, and this was recrystallized from ethanol to obtain 9.86 g of the target compound (yield 69.6%).
I got it.

Ultraviolet absorption spectrum (UV)i (in methanol)
: 235 nm (neutral) Elemental analysis: Calculated value as Cx, s Ho, N, O, engineering C, 37,74%; H, 3,35%; N, 8.8
1% actual value C, 37,76%; H, 3,18%; N,
8.74% Melting point (m, p,) = 139-141°C
Reference example 2 (1) 5-trimethylsilylethynyl-1-(2°3.
Synthesis of 5-tri-O-acetyl-β-D-ribofuranosyl)imidazole-4-carbonitrile 5-iodo-1-(2,3,5-tri-O-acetyl-β-D-ribofuranosyl)imidazole obtained in Reference Example 1 477 cm of -4-carbonitrile and 18 cm of bis(benzonitrile)palladium dichloride (5 mo 1%) were placed in a sealed tube, argon gas was bubbled through the tube, and the tube was dissolved in acetonitrile. This was added with triethylamine 0.16rnQ (1,
2 mmol) and 1.2+ mol of trimethylsilylacetylene (trimethylsilylethine) were added and reacted for 16 hours in an oil bath at 100°C. After the reaction, the first reaction solution was filtered through Celite, washed with ethanol, and purified with a silica gel column. The target compound (yield 76.4%) was obtained.

Mass spectrum (MS): 447 (molecular ion beak (M+)) 1H-nuclear magnetic resonance spectrum (NMR) (CDCI
,)2.12 (s,3HX3.acetyl)0.30
(s, 9H, trimethylsilyl) (2) 5-ethynyl-
Synthesis of 1-β-D-ribofuranosylimidazole-4-carbonitrile The compound obtained in (1) above was mixed with ammonia-
After one reaction in which it was dissolved in a mixed solvent of methanol and reacted at room temperature for 6 hours, the solvent was distilled off, and the resulting residue was purified with a silica gel column to obtain the target compound almost quantitatively.

MS: 249 (M+)”H-NMR (DMSO-d,): 5.35 (s
, LH, ethynyl) Reference Example 3 Synthesis of 5-alkynyl-1-β-D-ribofuranosylimidazole-4-carbonitrile Using the compound obtained in Reference Example 1 as a raw material, acetylene derivatives (phenylethyne, 3-
Hydroxy-1-propyne, 1-hexyne) in Reference Example 2
The target compound corresponding to the acetylene derivative was obtained by the same reaction. Identification data of the obtained compounds are shown in Table 1.

In Table 1, the yield indicates the yield from the triacetyl compound obtained in the intermediate stage.

Reference Example 4 Synthesis of 5-ethynyl-1-β-D-ribofuranosylimidazole-4-carboxamide (1) 5-(2-trimethylsilyl-1-ethyne-1
Synthesis of -1-(2,3,5-tri〇-acetyl-β-D-ribofuranosyl)imidazole-4-carboxamide 5-iodo-1-(2,3,5-doly) obtained in Reference Example 1 〇-acetyl-β-D-ribofuranosyl)imidazole-4-carboxamide 990■ (2 mmol) and bis(
benzonitrile) palladium dichloride 36Hg (0
, 1 mmol; 5 mo 1%) was placed in a sealed tube, argon gas was bubbled through it, and it was dissolved in 7 mQ of acetonitrile. To this was further added 850 μm (2.2 mmol) of trimethyl[(tributylstannyl)ethynylcosilane, and
After 17 hours of reaction in an oil bath at 0.degree. C., the reaction solution was filtered through Celite, washed with ethanol, and the solvent was distilled off. The obtained residual liquid was passed through a silica gel column (2.7X18cs
+) and eluted with hexane-ethyl acetate to obtain 668 ml of a candy-like substance (yield 71.8%).

UV (in methanol); λwax 270nm, 269nm (acidic), 2
61 nm (basic) MS: 465 (M") "H-NMR (CDCI, ): 2.17, 2.11, 2.10 (s, each 3H,
acetyl), 0.30 (s, 9H, trimethylsilyl) (2) Synthesis of 5-ethynyl-1-β-D-ribofuranosylimidazole-4-carboxamide 400 μ of the candy-like substance obtained in (1) above was treated with ammonia. - Dissolved in a mixed solvent of methanol and left overnight, then distilled off the solvent, adsorbed the resulting residue on a silica gel column, eluted with a 5-10% ethanol-chloroform mixed solvent, and crystallized from ethanol to form white crystals. 200 mg (yield 87.1%) was obtained.

m, p,: 182-185℃ Elemental analysis: Calculated value as C□, H□, N□O6 C, 4
9,44%; H, 4,90%; N, 15.72%'M 噸 C, 49,45%: H, 4,95%; N, 15.62
%MS 8 267 (M+) "H-NMR (DMSO-d,): 4.86 (s, LHl-C=CH) Reference Example 5 Synthesis of 5-iodoimidazole-4-carbonitrile 5-aminoimidazole-4 -Carbonitrile 3, 24
．． g (30.0 mmol) was dissolved in short methane 150 mA in a 100°C oil bath, and further, isoamyl nitrite 15- was added and reacted for 30 minutes.

After the reaction, the reaction solution was adsorbed on a silica gel column, eluted with ethanol-chloroform, and the fraction containing the target compound was concentrated to obtain 4.50 g of powder of the target compound (yield 70.0%).
I got it.

Elemental analysis: C, H, N, I as T1 value C, 21,94%; H, 0,92%;
N, 19. 19% 3, ti ma 41 f direct C, 2
2.15% iH, 0.77%; N, 19. 08
% Example 1 Synthesis of 5-ethynylimidazole-4-carbonitrile [-Formula [1]: A=CN, R1=Hydrogen atom] 5-trimethylsilylethynyl obtained in Reference Example 2 (1)
1-(2,3,5-tri〇-acetyl-β-D-ribofuranosyl)imidazole-4-carbonitrile 477
mg (1 mmol) IN hydrochloric acid-methanol (lo
After the reaction, the mixture was dissolved in a mixed solvent of d-5mM) and heated under reflux at 100° C. for 4 hours. The reaction solution was neutralized with IN sodium hydroxide solution, silica gel powder was added, and the solvent was distilled off. The resulting residue was filtered through a silica gel column (2.7x13
ao), eluted with a 0-8% ethanol-chloroform mixed solvent, and crystallized from aqueous methanol to obtain 5-
112 square meters (yield: 95.7%) of pale yellow crystals of ethynyl imidazole-4-carbonitrile were obtained.

m, p,: 169°C Elemental analysis: Calculated values as C, H, N3 C, 61,53%; H, 2,58%; N, 35
．． 88% Kenshin C, 61,50%; H, 2,52%; N,
35.78% UV (H, peak: λwax 254nm (neutral) 254nn+ (acidic) 254nm (basic) MS: 117 (M”) J (-NMR (DMSO-d, ): 13.7 (bs, LH , H-1), 7.99 (s,
IH, H-2), 5.02 (s, LH, CECH) Example 2 Synthesis reference example 4 of 5-ethynyl imidazole-4-carboxamide [general formula (I): A=CONH2, R''=hydrogen atom] 200 mg of 5-ethynyl-1-β-D-ribofuranosylimidazole-4-carboxamide (
0.75 mmol) was dissolved in IN hydrochloric acid solution 5-
The mixture was heated under reflux at 00°C for 4 hours. After reaction, 5 reaction solutions were added
The mixture was neutralized with sodium hydroxide solution, silica gel powder was added, and the solvent was distilled off.

The obtained residue was applied to a silica gel column (2,7X11a1).
The mixture was adsorbed onto the solution, eluted with a 0-16% ethanol-chloroform mixed solvent, and crystallized from an ethanol-hexane mixed solvent to give 38 cm of pale yellow crystals of 5-ethynyl imidazole-4-carboxamide (yield 37.6%). Obtained.

m, P,: 208-209°C (decomposition) Elemental analysis: Calculated values as C, HsN, O C, 53,33%; H, 3,73%; N, 31
．． 10% real o ■ direct C, 53, 18%-; H, 3,66
%; N, 31.15% UV (H, output: λll1ax 262 nm (neutral) 249 nm (acidic) 279 nm (basic) MS: 135 (M+) "H-NMR (DMSO-dG): 13,0 (bs , LH,H-1) , 7.71(
s, IH, H-2), 7.25, 7.22 (bs, 2
H, Nl-l2), 4.52 (s, LH, C
=CH) Example 3 5-Alkynylimidazole [General formula (1): A=CN, R1=phenyl group.

Synthesis of hydroxymethyl group, butyl group] The compound obtained in Reference Example 3 was treated in the same manner as in Example 1 to obtain a 5-alkynylimidazole having an alkynyl group corresponding to the starting compound.
4-Carbonitrile was obtained. Identification data of the obtained compounds are shown in Table 2.

Example 4 5-alkynylimidazole-4-carboxamide [-
Synthesis of formula (1): A=CONH2,R"=phenyl group, hydroxymethyl group, butyl group] The compound (0.03 mol) obtained in Example 3 was mixed with ammonia water-methanol (1
: 1) Dissolved in 30 ml of mixed solvent, added 3 mfi of hydrogen peroxide solution, and reacted at room temperature for 45 minutes. After the reaction, 5-alkynylimidazole-4-carboxamide having an alkynyl group corresponding to the raw material compound was obtained by purification with a silica gel column.

Example 5 Synthesis of 5-ethynylimidazole-4-carbonitrile (1) 5-iodoimidazole-4- obtained in Reference Example 5
220 mg of carbonitrile and 18 mm of bis(benzonitrile) palladium dichloride (5 mo 1%) were placed in a sealed tube, argon gas was bubbled through the tube, and the tube was dissolved in acetonitrile. To this, triethylamine 0,16mQ (1,2mmo
1) and 1.2 m+++ol of trimethylsilylacetylene (trimethylsilylethyne) were added, and the mixture was reacted in an oil bath at 100°C. After the reaction, the reaction solution was filtered through Celite, washed with ethanol, and purified with a silica gel column.
5-(2-trimethylsilyl-1-ethyn-1-yl)
Imidazole-4-carbonitrile was obtained.

(2) The compound obtained in (1) above was directly dissolved in a mixed solvent of ammonia-methanol, and reacted at room temperature for 6 hours. After the reaction, the solvent was distilled off, and the resulting residue was purified using a silica gel column and crystallized from aqueous methanol to obtain 5-ethynylimidazole-4-carbonitrile 12
2■ (yield: 65%) of crystals were obtained.

Example 6 Synthesis of 5-ethynylimidazole-4-carboxamide 5-ethynylimidazole-4-carbonitrile 117
(1.0 mmol) was dissolved in IN sodium hydroxide-methanol (1:1) mixed solvent 6III base.

0.1 mQ of hydrogen peroxide solution was added to this, and the mixture was stirred and reacted at room temperature for 45 minutes. After the reaction, the solvent was distilled off, the residual liquid was adsorbed on a silica gel column, eluted with a 0 to 16% ethanol-chloroform mixed solvent, and crystallized from an ethanol-n-hexane mixed solvent to obtain 5-ethynyl imidazole-4.
-Crystals of carboxamide were obtained.

Example 7 Tablet 5-ethynylimidazole-4-carbonitrile 10g Cornstarch 65g Carboxymethylcellulose 20g Polyvinylpyrrolidone
3g calcium stearate
2g total amount
100 g Prepare 100 square tablets using a conventional method. One tablet contains 10 ml of 5-ethynylimidazole-4-carbonitrile.

Example 8 Powder, capsule 5-ethynylimidazole-4-carboxamide 20g crystalline cellulose sog total amount
Mix 100g of both powders to make a powder. Further, 100 μm of the powder was filled into No. 5 hard capsules to prepare capsules.

Patent applicant (677) Yamasa Soy Sauce Co., Ltd. Sumitomo Pharmaceutical Co., Ltd.

Claims (1)

[Claims] 1) General formula [I] ▲There are mathematical formulas, chemical formulas, tables, etc.▼[I] [In the formula, A represents a carbamoyl group or a cyano group, and R
^1 represents a hydrogen atom, a lower alkyl group, a hydroxy lower alkyl group, or a phenyl group. ] An imidazole derivative or a salt thereof. 2) An antitumor agent comprising the imidazole derivative according to claim 1 or a pharmaceutically acceptable salt thereof as an active ingredient.