NO178304B - Intermediate for the preparation of pyrrolopyrimidine derivatives - Google Patents

Abstract

wherein ring A is a pyrrole or pyrroline ring, X is an amino group or a hydroxyl group, Y is a hydrogen atom, an amino group or a hydroxyl group ,. R is a hydrogen atom, a fluorine atom, an alkyl group, an alkenyl group or an alkynyl group, -COOR is a carboxyl group which may be esterified and n is a number of 2 to 4, and R may be different 1 each of the n repeating or a salt thereof , is described.

Description

The present invention relates to intermediates for the production of new pyrrolopyrimidine derivatives.

Folic acid is a carrier of a Cl unit in the body, derived from formic acid or formaldehyde, acting as a coenzyme in various enzymatic reactions such as in the biosynthesis of nucleic acids, in the metabolism of amino acids and peptides and in the formation of methane. Particularly in the biosynthesis of nucleic acids, folic acid is essential for formulation in the two reaction pathways, i.e. the purine synthetic reaction pathway and the thymidine synthetic reaction pathway. Generally, folic acid is required to be converted to the activated coenzyme form by reduction in two steps before it becomes biologically active. Amethopterin (methotrexate: MTX) and related compounds are known to inhibit the reduction from dihydrofolic acid to tetrahydrofolic acid by strongly binding to the dominant enzyme in the second step (dihydrofolic acid reductase). These drugs have been developed as anti-tumor drugs because they can interfere with DNA synthesis and thus cause cell death, and are currently considered to be clinically important. In contrast, a new tetrahydroaminopterin antitumor agent (5,10-dideaza-5,6,7,8-tetrahydroaminopterin: DDATHF) has been reported which, unlike the drugs described above, does not inhibit dihydrofolic acid reductase and the main mechanism consists in the inhibition of glycinamide ribonucleotide transformylase which is necessary in the initial phase of purine biosynthesis (Journal of Medicinal Chemistry, 28, 914 (1985)).

Various studies on the therapy of cancer have now been carried out, and the development of drugs that are more effective and have toxicities with high specificity towards cancer cells based on a new mechanism is expected. The anti-tumour agent MTX, in which the mechanism of action consists in antagonism to folic acid, is widely used clinically, despite the fact that the therapeutic effect is still unsatisfactory, due to its relatively strong toxicity with little effect on solid cancer.

As a result of the inventors' research under the conditions described above, they have discovered that new pyrrolopyrimidine derivatives have toxicities that are very specific to tumor cells and with good anti-tumor effects.

The new therapeutically active pyrrolopyrimidine derivatives have formula (I)

wherein ring Å is a pyrrole or pyrroline ring, X is an amino group or a hydroxyl group, Y is a hydrogen atom or a hydroxyl group, B is a straight or branched alkyl of 2-4 C atoms and —COOR<1> and —COOR^ are carbonyl groups which can be esterified with a C1-C4 alkyl or benzyl group or salts thereof, The compounds of formula I or salts thereof are prepared by a compound of formula (II)

wherein ring Å is a pyrrole or pyrroline ring, X is an amino group or a hydroxyl group, Y is a hydrogen atom or a hydroxyl group, a reactive derivative at the carboxyl group or a salt thereof, and a compound of formula

(III)

in which —COOR<1> and -C00R<2> are carboxyl groups which can be esterified with a C1-C4 alkyl or benzyl group or salts thereof are reacted. The present invention therefore relates to an intermediate product with formula

characterized in that ring A is a pyrrole or pyrroline ring, X is an amino group or a hydroxyl group, Y is a hydrogen atom or a hydroxyl group, B is a straight-chain or branched alkylene with 2-4 C atoms and R<3> is a hydrogen atom or a linear or branched alkyl of 1-4 carbon atoms, or a salt thereof.

When X or Y in the formulas described above is a hydroxyl group, each of the compounds (I) and (II) may exist as an equilibrium mixture of the respective tautomers. The following partial structural formulas show the sites on the structure that are subject to tautomerism, and the equilibria between the tautomers are illustrated below.

For simplicity of description, only the hydroxyl forms and the corresponding names are described in this specification, but the corresponding oxo forms are always included.

Alkyl groups represented by R in the formulas described above include alkyl groups of 1 to 4 carbon atom(s) each (eg methyl, ethyl, propyl, isopropyl groups). The alkyl groups include methyl, ethyl, propyl, isopropyl, n-butyl, iso-butyl, sec-butyl, tert-butyl, n-pentyl, iso-pentyl, sec-pentyl, neo-pentyl and tert-pentyl.

In the following, the method for producing starting compound (II) is explained.

Compound (II) in which ring A is a pyrrole ring can, for example, be prepared by the following methods. In the reaction formula described above, X, Y and R<3> are the same as described before; R 1 , R<b> and R<c> are independently a hydrogen atom, a fluorine atom or an alkyl group (same as those representing R described above); R<4> is a cyano group or an esterified carboxyl group represented by formula —COOR<5>; A is a hydrogen atom or a halogen atom (eg, fluorine atom, chlorine atom, bromine atom, iodine atom); B is a halogen atom (eg chlorine atom, bromine atom, iodine atom) or an eliminable group which can be easily derived from the hydroxy group (eg methanesulfonyloxy group, benzenesulfonyloxy group, p-toluenesulfonyl group, trifluoromethanesulfonyloxy group); and m is 0, 1 or 2. R<5> in the esterified carboxyl group represented by formula —COOR<5> is exemplified by an alkyl group of 1 to 4 carbon atom(s) (e.g. methyl, ethyl, propyl, isopropyl , butyl, sec-butyl, tert-butyl, etc.), phenyl or substituted phenyl group (p-nitrophenyl, p-methoxyphenyl, etc.), and benzyl or substituted benzyl (e.g. p-nitrobenzyl, p-methoxy- benzyl, etc. ).

Compound (V) can be dehydrated at the possible position between the two adjacent carbons and form an unsaturated bond.

In the following, the reaction modes described above are explained in detail.

Procedure 1

Compound (V) and compound (VI) are subjected to condensation and the resulting product is subjected to reduction to give compound (VII).

For the condensation, a known reaction (e.g. aldol reaction, Reformatsky reaction, Wittig reaction, etc.) can be used, and for the reduction usually a catalytic reduction under a hydrogen atmosphere in the presence of a catalyst (e.g. nickel, palladium, platinum, rhodium) advantageously used.

In the condensation of the aldol reaction, the useful basic catalysts include metal hydroxides such as sodium hydroxide, potassium hydroxide, lithium hydroxide and barium hydroxide, metal alkoxides such as sodium methoxide, sodium ethoxide and potassium tert-butoxide, metal amides such as sodium amide and lithium diisopropylamide, metal hydrides such as sodium hydride and potassium hydride, organic metal compounds such as phenyllithium and butyllithium and amines such as triethylamine , pyridine, α-, β- or β-picoline, 2,6-lutidine, 4-dimethylaminopyridine, 4-(1-pyrrolidinyl)pyridine, dimethylaniline and diethylaniline; useful acid catalysts include mineral acids such as hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid and boric acid, and organic acids such as oxalic acid, tartaric acid, acetic acid, trifluoroacetic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid and camphorsulfonic acid. Condensation can be carried out according to the known method [Ei-Ichi Negishi, Organometallics in Organic Synthesis, vol. 1, John Wiley & Sons, New York, Chichester, Brisbane, Toronto (1980)] which involves conversion of a ketone form to the silyl enol ether form, which is then subjected to condensation with an aldehyde or equivalent in the presence of a Lewis acid (e.g. eg anhydrous zinc chloride, anhydrous aluminum chloride (AICI3), anhydrous ferric chloride, titanium tetrachloride (TiC^), tin tetrachloride (SnC14), antimony pentachloride, cobalt chloride, cupric chloride, boron trifluoride ethyl ether complex, etc.), or conversion of a ketone form to the enolate by treating the ketone form with a metal triflate (e.g. dialkyl or boron or tin(II) triflate) in the presence of amines (e.g. triethylamine, pyridine, a-, e- or "y-picoline, 2,6-lutidine, 4-dimethylaminopyridine, 4-(1-pyrrolidinyl)pyridine, dimethylaniline, diethylaniline) followed by subjecting the enolate to condensation with an aldehyde or equivalent. The condensation is carried out in a suitable solvent at a temperature varying from -100°C to the boiling point of the solvent, insinuate varying from -78 to 100°C, for 1 minute to 3 days. Solvents that can be used in the reaction include water, liquid ammonia, alcohols (e.g. methanol, ethanol, propanol, isopropanol, butyl alcohol, sec-butyl alcohol, tert-butyl alcohol, ethylene glycol, methoxyethanol, ethoxyethanol), ether compounds (e.g. dimethyl ether , diethyl ether, tetrahydrofuran, dioxane, monoglyme, diglyme), halogenated hydrocarbons (e.g. dichloromethane, chloroform, carbon tetrachloride), aliphatic hydrocarbons (e.g. pentane, hexane, heptane), aromatic hydrocarbons (e.g. benzene, toluene , xylene), dimethylformamide, dimethylsulfoxide, hexamethylphospholamide, sulfolane, and suitable mixtures thereof. In the condensation of the Wittig reaction, useful reagents include metal hydroxides such as sodium hydroxide, potassium hydroxide, lithium hydroxide and barium hydroxide, metal alkoxides such as sodium methoxy, sodium ethoxide and potassium tert-butoxide, metal amides such as sodium amide and lithium diisopropylamide, metal hydrides such as sodium hydride and potassium hydride, organic metal compounds such as phenyllithium and butyllithium, and amines such as triethylamine, pyridine, a-, p- or "y-picoline, 2,6-lutidine, 4-dimethylaminopyridine, 4-(1-pyrrolidinyl)pyridine, dimethylaniline and diethylaniline. The reaction is carried out in a suitable solvent at a temperature varying from -20°C to the boiling point of the solvent being used, preferably varying from 0 to 150°C, for 1 minute to 10 days. Solvents which can be used in the reaction include liquid ammonia, alcohols (e.g. e.g. methanol, ethanol, propanol, isopropanol, butyl alcohol, sec-butyl alcohol, tert-butyl alcohol, ethylene glycol, met oxyethanol, ethoxyethanol), ether compounds (e.g. dimethyl ether, diethyl ether, tetrahydrofuran, dioxane, monoglyme, diglyme, aliphatic hydrocarbons (e.g. pentane, hexane, heptane), aromatic hydrocarbons (e.g. benzene, toluene, xylene), dimethylformamide, dimethylsulfoxide, hexamethylphospholamide, sulfolane and suitable mixtures thereof.

The condensation can also be carried out using a Reformatsky reaction. Reagents useful in the Reformat cloud reaction include zinc, magnesium, aluminum and tin, and the reaction is carried out in a suitable solvent at a temperature ranging from -20°C to the boiling point of the solvent being used, preferably ranging from 0 to 150°C , for 30 minutes to 3 days. The solvents which can be used in the reaction include ether compounds (eg dimethyl ether, diethyl ether, tetrahydrofuran, dioxane, monoglyme, diglyme), aliphatic hydrocarbons (eg pentane, hexane, heptane), aromatic hydrocarbons (eg benzene , toluene, xylene) and suitable mixtures thereof.

The reaction conditions for the catalytic reduction are the same as the esterification by —COOR<1> and —COOR<2> of compound (III) (method C).

The starting materials (V) and (VI) can easily be provided according to the known methods described in the literature.

(B. Neises et al., Angew. Chem. Int. Ed. Engl., 17, 522

(1978)).

Procedure 2

This is the method by which an eliminable functional group B is introduced into the active methyl (a-position of the carboxylic acid ester compound) in compound (VII) and can be easily carried out using known reagents according to a method known per se.

Procedure 3

Compound (VIII) obtained in method 2 is subjected to condensation with malononitrile or a cyanoacetic acid ester [NC-CH2COOR<5>; R<5> is as described above] under basic conditions, to give compound (IX). The applicable bases, solvents and reaction conditions are according to the known methods.

Procedure 4

Compound (IX), when treated with guanidine, can react at the cyano group or the ester residue followed by cyclization to form a pyrrolopyrimidine ring. The ring closure under basic conditions means that the reaction proceeds advantageously. Useful bases include metal alkoxides, such as sodium methoxide, sodium ethoxide or potassium tert-butoxide. Useful solvents for the reaction include methanol, ethanol, propanol, tert-butyl alcohol, dimethylsulfoxide and hexamethylphospholamide. The reaction temperature varies from 0 to 150°C, preferably from 20 to 100°C. The reaction time varies from 1 to 48 hours.

Procedure 5

Compound (IV-1: Y=NH2 or OH) obtained in method 4 can be converted to compound (II-1: Y=NE2 or OH) by subjecting the ester residue [—COOR<3>] to esterification which was used for preparation of compound (I-l).

Procedure 6

Compound (II-1: Y=OE) provided in 1 method 5 was subjected to reduction to give compound (II-2: Y=H). The conditions for the reduction are known per se, and reduction by means of a metal hydride (e.g. borane, alane or at-complex thereof) is advantageously used.

Method 5 and method 6 can be carried out in reverse order. That is, in method 7, compound (IV-1: Y=OH) is subjected to reduction as in method 6 to give compound (IV-2: Y=H), which is then subjected to esterification in method 8 on a similar manner as in method 5 to give compound (II-2: Y=E). Either the esterification or the reduction can be chosen to be carried out first relative to the other according to the nature of the substituents in compound (IV-1: Y=0E).

In methods 6 and 8, the mixture containing the compounds (II-2) and (II-2') or the compounds (IV-2) and (IV-2') can be separated, or each of the compounds (II-2) and ( II-2') or each of the compounds (IV-2) and (IV-2') are prepared mainly by selective reduction.

Among the compounds (II), those represented by formula (II-2: X=OH)

wherein R and n mean the same as described above, also be provided by the following methods In the methods described above, R, R<3>, Y and n mean as described before and Z means formula RCH2CO— in which R means the same as described above , formula in which L is phenyl, butyl or cyclohexyl, and R and n mean the same as described above, or formula

wherein M is ethyl or phenyl, and R and

n means the same as described above. It is preferred that Y is hydrogen.

In the following, these methods are explained.

Procedure 9

Compound (X) [T. Kondo et al., Chemistry Letters, 419

(1983) and a para-substituted benzoic acid ester derivative (XI) is subjected to condensation (aldol reaction, Wittig reaction) followed by catalytic reduction under a hydrogen atmosphere, which gives compound (XII). In the condensation, the same reaction conditions, reaction solvents, reaction temperatures and reagents are used as in method 1. For the catalytic reduction under a hydrogen atmosphere, the conditions used in the deesterification of —COOR<1> and —COOR<2> of compound (III) are applicable .

Procedure 10

Treatment of compound (XII) under acidic conditions can eliminate the protection of the isopropyloxymethyl group at the 3-position to give compound (XIII). The conditions, the solvents and the temperatures used in the removal of -COOR<1> and -COOR<2> of compound (III) (method B-1 and method B-2) applicable in the reaction.

Procedure 11

Compound (XIII) obtained in method 10 is subjected to dehydration by a method known per se, to be easily converted into compound (IV-3: Y=H).

Procedure 12

Compound (IV-3: Y=H) provided in method 11 can be converted to compound (II-3) by deesterification. The conditions, solvents, and temperatures described in detail for the removal of -COOR<1> and -COOR<2> of compound (III)

(methods A, B-1, B-2 and C) can be used in the reaction. The processes 10 and 12 can be carried out in any order with the formation of the respective products, and finally the desired compound (II-3) is provided. The order is suitably determined according to the nature of the substituents of the compounds (XII), (XIII) and (IV-3). Compound (II-3) thus obtained can be converted, if necessary, into compound (II-2) by means of a known substituent-converting reaction on the pyrimidine ring indicated in the literature. [Protein Nucleic acid Enzyme Extra Issue, Chemical synthesis of nucleic acids, Kyoritsu Shuppan (1968)].

Compounds other than compound (II-3) in which X is hydroxyl can also be converted into the corresponding compounds in which X is amino by means of the above-mentioned substituent-converting reaction.

The reactions, reagents and reaction conditions used in processes 1 to 12 and for the production of starting compounds (V) and (XIII) are known and explained in detail in the following literature.

[J.F.W. Mcomine, Protective Groups in Organic Chemistry, Plenum Press, London and New York (1973)], [Pine, Hendrikson, Hammond, Organic Chemistry (4th ed.) [I]-[II], Hirokawa Shoten (1982)], and [M. Fieser and L. Fieser, Reagents for

Organic Synthesis vol. 1-10, Wiley-Interscience, New York, London, Sydney and Toronto (1969-1982)].

The products of this invention can be isolated from the reaction mixtures using conventional methods of separation and purification, such as concentration, solvent extraction, chromatography and recrystallization.

The compounds (I) according to this invention can form salts. Such salts are prepared using the known methods, and are exemplified by the salts of the bases or acids and the quaternary salts. Salts of the bases include salts of alkali metals, alkaline earth metals, non-toxic metals, ammonia and ammonia preferably substituted with Cl_3 alkyl or ethanol, such as sodium, potassium, lithium, calcium, magnesium, aluminum, zinc, ammonium, trimethylammonium, triethylammonium, triethanolammonium nium, pyridinium and pyridinium preferably with C1-3 alkyl or ethanol. The salts of the acids include salts of mineral acids such as hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid and boric acid, and salts of organic acids such as oxalic acid, tartaric acid, acetic acid, trifluoroacetic acid, methanesulfonic acid and camphorsulfonic acid. Quaternary salts include salts of methyl bromide, methyl iodide, methyl methanesulfonate, methylbenzenesulfonate and methyl p-toluenesulfonate.

The compounds (I) and (II) can also form zwitterions.

The following reference examples and working examples will explain the present invention in more detail.

Reference example 1

Preparation of methyl 5-[4-(tert-butoxycarbonyl)-phenyl]pentanoate: Under an argon atmosphere, potassium (25 mg) was added to dried tert-butyl alcohol (820 ml), which was refluxed to complete dissolved. The solution was cooled to 20°C, whereupon ether (300 mL) was added followed by a solution of methyl crotonate (63.93 g) and tert-butyl 4-formylbenzoate (71.0 g) in tert-butyl alcohol ether ( 2:1, 300 ml) was slowly added while maintaining the internal temperature at 10°C. After stirring at the same temperature for 2 hours, 1 N potassium hydrogen sulfate in water (750 mL) was added with cooling to adjust the pH to 4. The solution was extracted with ether, washed with water and then with saturated sodium chloride solution and subjected to evaporation of the solvent under reduced pressure. The resulting residue was dissolved in ethyl acetate (100 ml), and 5#Pd-C (15 g: Engelhard Co. Ltd.) was added and extensively stirred under a hydrogen pressure of 4 kg/cm<2> at room temperature for 3 hours. The catalyst was filtered out, the solvent was evaporated under reduced pressure, and dried methanol (200 mL), 4-(N,N-dimethylamino)pyridine (30 mg) and dichloromethane (250 mL) were added to the residue and then a solution consisting of 1,3-dicyclohexylcarbodiimide (132 g) in dichloromethane (250 ml) slowly added dropwise at 0°C. After stirring at room temperature for 18 hours, the mixture was cooled to 0°C, and acetic acid (30 mL) was added and the mixture was stirred at 0°C for 30 minutes and then at room temperature for 30 minutes. The resulting precipitate was filtered off, the filtrate was concentrated to dryness under reduced pressure, and to the residue ethyl acetate (100 ml) was added and after standing at 0°C for 2 hours the resulting precipitate was again filtered out. The filtrate was concentrated under reduced pressure and the residue was purified by column chromatography (carrier ; silica gel, 100 g, eluent; ether: hexane = 1:15 1:5), to give the final compound (59.7 g).

Melting point (Bp) 145-1550 C/0.2-0.3 mmHg

IR (Neat): 2980, 2950, 1740, 1712, 1605 cm-<1>.

1-H-NMR (CDCl 3 ) S: 1.40-1.75 (4H,m), 1.55 (9H,s), 2.15-2.45 (2E,m), 2.50-2 .75 (2H,m), 3.62 (3E,s),

7.16 (2H,d,J=8Ez), 7.85 (E,d,J=8Ez).

Reference example 2

Preparation of methyl 5-[4-(tert-butoxycarbonyl)-phenyl]-2-iodopentanoate: Under an argon atmosphere, a solution of diisopropylamine (2.48 g) in tetrahydrofuran (100 ml) was added to a solution of butyllithium ( 24.5 mmol) in hexane (15.3 ml) at 0°C and stirred for 10 minutes, to which a solution of the compound (6.53 g) provided in reference example 1 in tetrahydrofuran (50 ml) was added dropwise at - 78°C within 30 minutes. After stirring for 30 minutes, a solution of iodine (5.66 g) in tetrahydrofuran (30 ml) was added and stirred for a further 20 minutes. The temperature of the solution was brought up to 0°C within 30 minutes, 1 N potassium hydrogen sulfate in water (30 ml) was added dropwise, and the solution was extracted with ether after adjusting to pH 4. The organic layer was washed with 1 N potassium carbonate in water and then with saturated sodium chloride solution, and dried with anhydrous magnesium sulfate.The residue obtained by evaporation of the solvent under reduced pressure was purified by column chromatography (ether-hexane, 1:9) to give the final compound (4.736 g).

IR (Neat): 2990, 2905, 1744, 1718, 1612 cm-<1>.

1-H-NMR (CDCl 3 ) S: 1.45-1.80 (2H,m), 1.58 (9H,s), 1.80-2.16 (2H,m), 2.69 (2H ,t,J=7Hz), 3.72 (2H,s),

4.30 (lH,t,J=7Hz), 7.20 (2H,d,J=8Hz),

7.90 (2H,d,J=8Hz).

Reference example 3

Preparation of methyl 5-[4-(tert-butoxy-carbonyl)phenyl]-2-(dicyanomethyl)pentanoate: To a suspension of sodium hydride (1.356 g) in dimethyl sulfoxide (8 ml) was added a solution of malonitrile (3, 37 g) in dimethylsulfoxide (8 ml) while cooling with water, and stirred for 15 minutes. To this solution, a solution of the compound (4.736 g) provided in reference example 2 in dimethyl sulfoxide (12 ml) was added dropwise and stirred at room temperature for 1 hour, to this solution 45 ml of 1N potassium hydrogen sulfate in water was added at 0 °C followed by distraction with ether. The ether layer was washed with water and dried with anhydrous magnesium sulfate, followed by evaporation of the solvent under reduced pressure. The residue was purified by column chromatography (support; silica gel, 200 g, eluent; ethyl acetate; hexane = 1:5), to give the final compound (3.33 g).

IR (Neat): 2970, 2930, 2252 1740, 1713, 1608 cm-<1>

1H-NMR (CDCl 3 ) S: 1.60-2.05 (4H,m), 1.48 (9H,s),

2.70 (2H,brt,J=7Hz), 2.90-3.15 (lH,m), 3.82 (3H,s),

4.04 (lE,d,J=7Hz), 7.20 (2H,d,J=8Hz),

7.92 (2E,d,J=8Hz ).

Reference example 4

Preparation of methyl 4-[3-(2-amino-7-benzyl-3-isopropyloxy-methyl-4(3H)-oxopyrrolo[2,3-d]pyrimidin-5-yl)-1-oxo-2-propenyl ]benzoate: 2-Amino-7-benzyl-3-isopropyloxymethyl-4(3H)-oxo-pyrrolo[2,3-d]pyrimidine-5-carbaldehyde (1.7 g) was suspended in a methanol-tetrahydrofuran mixture (10 :1, 33 mL), whereupon a solution of sodium methylate in methanol (equivalent to 6.25 mM, 3.75 mL) was added to dissolve.

Then 4-methoxycarbonylacetophenone (2.23 g) was added and stirred at room temperature for 15 hours. The precipitate was collected by filtration, washed with a small amount of methanol and ether, and dried to give the final compound (2.02 g) as yellow needles.

IR (KBr): 3480, 3350, 1710, 1680, 1620, 1550,

1375, 1280, 1210, 1110, 1060, 775. cm-<1>.

<1>H-NMR (CDCl3) S: 1.23 (6H,d,J=6Hz), 3.93 (3E,s), 3.80-4.07 (1H,m), 5.15 ( 2H,s), 5.63 (2H,s),

6.92 (lH.s), 7.10-7.40 (5H, m), 7.73 (1H,d,J=15Hz),

8.13 (4H,s), 8.60 (1E,d,J=15Hz).

Reference example 5

Preparation of methyl 4-[3-(2-amino-3-isopropyloxymethyl-4(3H )-oxo-5,6-dihydropyrrolo-[2,3-d]pyrimidin-5-yl)propyl]-benzoate: The compound ( 2.01 g) provided in Reference Example 4 was dissolved in a methanol-tetrahydrofuran mixture (3:4, 350 ml), after which 1 N hydrochloric acid (8 ml) and 10% Pd-C (4 g, prepared by Engelhard Co. Ltd .) was added, and subjected to catalytic reduction under a hydrogen atmosphere for 48 hours. The catalyst was filtered out, the filtrate was neutralized, and the solvent was evaporated under reduced pressure and the residue was isolated and purified by column chromatography on silica gel, (carrier; 100 g, eluent; chloroform containing 2-4$ ethanol), to give final the parent compound (0.68 g) as a colorless powder.

IR (KBr): 3210, 2980, 1725, 1625, 1580, 1510,

1435, 1275, 1175, 1100, 1060 cm-<1>.

1-H-NMR (CDCl 3 ) S: 1.17 ( 3H , d , J=6Hz ), 1.19

(3H,d,J=6Ez ) , 1.50-2.13 (4H,m), 2.70 (2H,t,J=7.5Hz), 3.07-3.77 (3H,m) , 3.80-4.60 (lH,m), 3.87 (3H,s), 5.03 and 5.57 (2E,ABq), 7.21 (2E,d,J=7.5Ez) , 7.91

(2E,d,J=7.5Hz).

Reference example 6

Preparation of methyl 4-[3-(2-amino-4-hydroxy-5,6-dihydro-pyrrolo[2,3-d]pyrimidin-5-yl)propyl]benzoate: The compound (0.66 g) provided in reference example 5 was dissolved in dried tetrahydrofuran (31.5 ml), whereupon 0.21 N hydrogen bromide in dichloromethane (78.3 ml) was added and stirred at room temperature for 20 hours. Then 3 volumes of n-hexane were added and the resulting precipitate was collected by filtration to give the dihydrobromide of the final compound (0.59 g) as a colorless powder.

IR (KBr): 3290, 3030, 2950, 1720, 1690, 1680

1620, 1480, 1350, 1275, 1100, 1035, 760 cm-<1>.

iE-NMR (DMSO-d6) S: 1.40-1.83 (4E,broad),

2.65 (2H,t,J=7.5Hz) , 3.07-3.37 (2H,m), 3.50-3.77

(1H),m), 3.82 (3H,s), 7.33 (2H,d,J=7.5Ez),

7.86 (2H,d,J=7.5Hz).

Reference example 7

Preparation of diethyl N-[4-[3-(2-amino-4-hydroxy-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)propyl]benzoyl]-L-glutamate: The compound (1 .47 g) provided in reference example 6 was suspended in tetrahydrofuran (60 ml), whereupon 0.1 N sodium hydroxide in water (120 ml) was added and stirred at room temperature for 21 hours. The solution was then neutralized with 0.1 N hydrochloric acid (60 ml) and concentrated to dryness under reduced pressure. The residue was suspended in dried dimethylformamide (112.5 ml), whereupon diethyl L-glutamate hydrochloride (2.88 g), diphenylphosphoryl azide (1.295 ml) and triethylamide (2.52 ml) were added, returned to room temperature and stirring was continued for 63 hours. The resulting precipitate was filtered off, and the filtrate was concentrated to dryness under reduced pressure. The residue was subjected to separation purification by column chromatography on silica gel (support; 100 g, eluent; chloroform containing 6.9% ammonia-containing ethanol, 1:20 1:10), to give the final compound (1.12 g) as a colorless powder.

IR (KBr): 3330, 2930, 1740, 1670, 1640, 1570,

1540, 1440, 1375, 1300, 1200, 1095, 1020 cm-<1>.

1-H-NMR (CDCl 3 -CD 3 OD) S: 1.20 ( 3H, t, J=7 , 5Hz ),

1.27 (3E,t,J=7.5Hz), 1.47-1.83 (4H,m),

2.0-2.36 (2E,m), 2.37-2.50 (2H,m), 2.67

(2H,t,J=7.5Hz), 3.10-3.37 (2H,m), 3.53-3.80 (lH,m), 3.96-4.33 (4H,q x 2 ,J=7.5Hz), 4.60-4.87 (lH,m),

7.25 (2H,d,J=9Hz), 7.75 (2H,d,J=9Hz).

Reference example 8

Preparation of methyl 4-[3-(2-amino-7-benzyl-3-isopropyloxy-methyl-4(3H)-oxopyrrolo[2,3-d]pyrimidin-5-yl)-1-oxo-2- propenyl]benzoate: 2-Amino-7-benzyl-3-isopropyloxymethyl-4(3H)-oxo-pyrrolo[2,3-d]pyrimidine-5-carbaldehyde (1.7 g) was suspended in a methanol-tetrahydrofuran mixture ( 10:1, 33 mL), after which a solution of sodium methylate in methanol (equivalent to 6.25 mM, 3.75 mL) was added to dissolve. Then 4-methoxycarbonylacetophenone (2.23 g) was added and stirred at room temperature for 15 hours. The precipitate was collected by filtration, washed with a small amount of methanol and ether, and dried to give the final compound (2.02 g) as yellow needles.

IR (KBr): 3480, 3350, 1710, 1680, 1620, 1550, 1376,

1280, 1210, 1110, 1060, 775 cm-<1>

<1->H-NMR (CDCl 3 ) S: 1.23 (6H, d, J=6Hz ), 3.93 (3H,s), 3.80-4.07 (1E,m), 5.15 (2H,s), 5.63 (2E,s), 6.92

(lE,s), 7.10-7.40 (5H,m), 7.73 (1H,d,J=15Hz), 8.13 (4H,s), 8.60 (lE,d,J =15Hz).

Reference example 9

Preparation of methyl 4-[3-(2-amino-3-isopropyloxy-methyl-4(3H)-oxo-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)propyl]-benzoate: The compound (2.01 g) provided in Reference Example 8 was dissolved in a methanol-tetrahydrofuran mixture (3:4, 350 ml), after which 1 N hydrochloric acid (8 ml) and 10% Pd-C (4 g, prepared by Engelhard Co. Ltd.) was added, and subjected to catalytic reduction under a hydrogen atmosphere for 48 hours. The catalyst was filtered out, the filtrate was neutralized, the solvent was evaporated under reduced pressure, and the residue was isolated and purified by column chromatography on silica gel (support; 100 g) (eluent: chloroform containing 2-4% ethanol), to give the final compound (0.68 g) as a colorless powder.

IR (KBr): 3210, 2980, 1725, 1625, 1580, 1510, 1435,

1275, 1175, 1100, 1060 cm-<1>.

%-NMR (CDCl 3 ) S: 1.17 (3H, d , J=6Hz ),

1.19(3H,d,J=6Hz), 1.50-2.13 (4H,m), 2.70

(2H,t,J=7.5Ez) , 3.07-3.77 (3H,m), 3.80-4.06 (1H,m),

3.87 (3H,s), 5.03 and 5.57 (2H,ABq), 7.21

(2H,d,J=7.5Hz), 7.91 (2H,d,J=7.5Hz).

Reference example 10

Preparation of methyl 4-[3-(2-amino-4-hydroxy-5,6-dihydro-pyrrolo[2,3-d]pyrimidin-5-yl)propyl]benzoate: The compound (0.66 g) provided in reference example 9 was dissolved in dried tetrahydrofuran (31.5 ml), whereupon 0.21 N hydrobromic acid in dichloromethane (78.3 ml) was added, and stirred at room temperature for 20 hours. Then 3 volumes of n-hexane were added and the resulting precipitate was collected by filtration to give the dihydrobromide of the final compound (0.59 g) as a colorless powder.

IR (KBr): 3290, 3030, 2950, 1720, 1690, 1680,

1620, 1480, 1350, 1275, 1100, 1035, 760 cm-<1>. <i>H-NMR (DMS0-d6) §: 1.40-1.83 (4H, broad), 2.65 (2H,t,J=7.5Hz), 3.07-3.37 (2H ,m), 3.50-3.77 (lH.m),

3.82 (3H,s), 7.33 (2H,d,J=7.5Hz ) , 7.86

(2H,d,J=7.5Hz).

Reference example 11

Preparation of methyl 4-[2-(2-amino-7-benzyl-3-isopropyloxy-methyl-4(3H)-oxopyrrolo[2,3-d]pyrimidin-5-yl)-ethenyl]benzoate: For a suspension consisting of 2-amino-7-benzyl-3-isopropyl-oxy-methyl-4(3H)-oxopyrrolo[2,3-d]pyrimidine-5-carbaldehyde (2.04 g) in dried methanol (84 ml) was p-Methoxycarbonyl-benzyltriphenylphosphonium bromide (3.24 g) added and stirred. Then a solution of sodium methylate in methanol (equivalent to 6.6 mM on a sodium basis) was added and stirred at room temperature for 1.5 hours to give yellow needles. The needles were collected by filtration, washed with methanol and then with ether, and dried to give the final product (cis form, 1.49 g). The mother liquor was purified by column chromatography on silica gel (support: 100 g) (eluent: ethyl acetate-hexane, 1:4-»1:3), to give the cis-trans mixture of the final compound (0.9 g) as a yellow powder, cis- shape;

IR (KBr): 3340, 3220, 2980, 1715, 1690, 1625,

1600, 1530, 1430, 1280, 1175, 1105,

1060, 995 cm-<1>.

3H-NMR (CDCl3) δ: 1.20 (6H,d,J=6Hz), 3.87 (3H,s), 3.80-4.07 (1H,m), 5.14 (2H, s), 5.32 (2E,s), 5.60

(2H,s), 6.77 (1H,s), 7.10-7.37 (5H,m), 7.43 (2H,s),

7.50 (2H,d,J=9Ez), 7.95 (2H,d,J=9Hz).

Reference example 12

Preparation of methyl 4-[2-(2-amino-3-isopropyloxymethyl-4(3H )-oxo-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)ethyl]benzoate: The compound (1, 6 g) provided in Reference Example 11 was subjected to the same reaction as that in Reference Example 9 to give the final compound (0.62 g).

<1->H-NMR (CDCl3) S. 1.17 ( 3E, d , J=6Hz ) , 1.20

(3H,d,J=6Hz), 1.47-2.0 (lH,m), 2.10-2.43 (lH,m), 2.65 (2H,t,J=9Hz), 2 .97-3.60 (3H,m), 3.73-4.07 (lE.m),

3.90 (3H,s), 4.47 (2H,s), 5.30 (1E,d,J=12Hz), 5.60 (lH,d,J=12Hz), 7.13-7, 50 (7H,m), 7.92 (1H,d,J=9Ez).

Reference example 13

Preparation of methyl 4-[2-(2-amino-4-hydroxy-7H-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)ethyl]benzoate: The compound (1.25 g) provided in Reference Example 12 was subjected to the same reaction as Reference Example 10 to give the final compound (0.51 g).

IR (KBr): 3400, 3300, 2920, 1740, 1710, 1680, 1640,

1600, 1570, 1435, 1310, 1280, 1110, 1020

cm-1.

<i>H-NMR (DMS0-d6/D20) S: 1.53-2.27 (2H,m), 2.70 (2H,t,J=9Hz) , 3.00-3.26 (2H ,m), 3.47-3.63 (lH,m), 3.83 (3H,s), 7.35 (2H,d,J=9Hz), 7.85 (2H,d,J=9Hz ).

Reference example 14

Preparation of methyl 5-[4-tert-butoxycarbonyl)-phenyl]penta-noate: Under an argon atmosphere, potassium (25 g) was completely dissolved in dried tert-butyl alcohol (820 ml) by heating under reflux for 3 hours. The solution was cooled to 20°C, whereupon ether (300 mL) was added and then slowly a solution of methyl crotonate (63.93 g) and tert-butyl 4-formylbenzoate (71.0 g) in tert-butyl alcohol-ether mixture ( 2:1, 300 ml) while the internal temperature was maintained at 10°C.

The mixture was stirred at the same temperature for 2 hours, and 1 N potassium hydrogen sulfate in water (750 mL) was added with cooling to adjust the pH to 4. After extraction with ether, the ether layer was washed with saturated sodium chloride solution and the solvent was evaporated under reduced pressure . The residue was dissolved in ethyl acetate (100 ml), whereupon 5% Pd-C (15 g: manufactured by Engelhard Co. Ltd.) was added, and extensively stirred at room temperature under a hydrogen pressure of 4 kg/cm<2> in 3 hours. The catalyst was filtered out, the solvent was evaporated under reduced pressure, and dried methanol (200 ml), 4-(N,N-dimethylamino)pyridine (30 mg) and dichloromethane (250 ml) were added, after which a solution of 1 ,3-Dicyclohexylcarbodiimide (132 g) in dichloromethane (250 mL) was slowly added dropwise at 0°C. After stirring at room temperature for 18 hours, the mixture was cooled to 0°C. Acetic acid (30 mL) was added and the mixture was stirred at 0°C for 30 minutes and then at room temperature for 30 minutes. The resulting precipitate was filtered out, the filtrate was concentrated to dryness under reduced pressure, and ethyl acetate (100 mL) was added to the residue, which was left at 0°C for 2 hours, and the resulting precipitate was again filtered out. The filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography (support: silica gel, 500 g, ether-hexane, 1:15-+1:5), to give the final compound (59.7 g).

Bp. 145-155°C/0.2-0.3 mmHg.

IR (Neat): 2980, 2950, 1740, 1712, 1605 cm-<1>.

^E-NMR (CDCl 3 ) S: 1.40-1.75 (4H,m), 1.55 (9E,s ), 2.15-2.45 (2H,m), 2.50-2, 75 (2H,m), 3.62 (3H,s), 7.16 (2H,d,J=8Hz), 7.85 (H,d,J=8Hz).

Reference example 15

Preparation of methyl 5-[4-(tert-butoxycarbonyl)-phenyl]-2-iodopentanoate: Under an argon atmosphere, a solution of butyllithium (24.5 mmol) in hexane (15.3 mL) was added to a solution of diisopropylamine (2 .48 g) in tetrahydrofuran (100 ml) at 0"C and stirred for 10 minutes. To the resulting solution, a solution of the compound (6.53 g) provided in Reference Example 14 in tetrahydrofuran (50 ml) was added dropwise at -78 °C over 30 min. After stirring for 30 min, a solution of iodine (5.66 g) in tetrahydrofuran (30 mL) was added and stirred for an additional 20 min. The temperature was raised to 0 °C over 30 min , and 30 mL of 1N aqueous potassium hydrogen sulfate was added dropwise, and the pH was adjusted to 4, followed by extraction with ether. The organic layer was washed with 1 N aqueous potassium carbonate and then with saturated sodium chloride solution, and dried with anhydrous magnesium sulfate. The solvent was evaporated under reduced pressure, and the resulting r the ester was purified by column chromatography (support: silica gel, 100 g, ether-hexane, 1:9), to give the final compound (4.736 g).

IR (Neat): 2990, 2905, 1744, 1718, 1612 cm-<1>.

<1->H-NMR (CDCl 3 ) δ: 1.45-1.80 (2H,m), 1.58 (9H,s), 1.80-2.16 (2H,m), 2.69 (2H,t,J=7Hz), 3.72 (2H,s),

4.30 (lH,t,J=7Hz), 7.20 (2E,d,J=8Hz),

7.90 (2H,d,J=8Hz).

Reference example 16

Preparation of methyl 5-[4-(tert-butoxycarbonyl)-phenyl]-2-(dicyanomethyl)pentanoate: To a suspension of sodium hydride (1.356 g) in dimethyl sulfoxide (8 ml) was added a solution of malonitrile (3.37 g) in dimethyl sulfoxide (8 mL) added with ice-cooling and stirred for 15 min. To the solution, a solution of the compound (4.736 g) provided in Reference Example 15 in dimethyl sulfoxide (12 ml) was added dropwise, and the mixture was stirred at room temperature for 1 hour. Then 45 ml of 1 N potassium hydrogen sulfate in water was added at (<T>C, followed by extraction with ether. The ether layer was washed with water and dried with anhydrous magnesium sulfate. The residue obtained by evaporation of the solvent under reduced pressure was purified by column chromatography (support: silica gel, 200 g, ethyl acetate-hexane, 1:5), to give the final compound (3.33

g)<.>

IR (Neat): 2970, 2930, 2252, 1740, 1713 (1608 cm-<1>.

<i>H-NMR (CDCl 3 ) S: 1.60-2.05 (4H,m), 1.48 (9H,s),

2.70 (2H,brt,J=7Ez), 2.90-3.15 (lH.m), 3.82(3H,s), 4.04(lH,d,J=7Ez), 7, 20 (2H,d,J=8Ez), 7.92 (2H,d,J=8Hz).

Reference example 17

Preparation of ethyl 5-[4-(tert-butoxycarbonyl)-phenyl]hexanoate: In a solution of tert-butyl 4-acetylbenzoate (19.90 g) in a benzene-ether-tetrahydrofuran mixture (3:3:2, 200 ml) zinc (11.81 g) was suspended, after which ethyl 4-bromocrotonate (17.44 g) was slowly added with heating and stirring, and then iodine (about 20 mg) was added. The resulting mixture was refluxed in an oil bath (60-70°C) for 1 hour, then ethyl 4-bromocrotonate (3.00 g) was added, and the mixture was further refluxed for 15 minutes. After cooling at room temperature, the reaction mixture was added to water (500 ml), adjusted to pH 4.9 by the addition of acetic acid and extracted with ether. The extract was washed with 5% aqueous ammonia and dried with anhydrous magnesium sulfate.

The residue obtained by evaporation of the solvent under reduced pressure was purified by column chromatography (support; silica gel, 300 g, eluent; ethyl acetate-hexane=1:5), to give the final compound.

IR (Neat): 3480, 2975, 1720, 1650, 1605 cm-<1>.

3H-NMR (CDCl3) S: 1.20 (3H,t,J=7Hz), 1.53 (12H,s),

2.64 (2H,d,J=7Hz), 2.67 (lH.brs), 3.63 (3H,s), 4.08 (2H,q,J=7Hz), 5.80 (1H, d,J=15Hz), 6.80

(lH,dt,J=15Hz,7Hz), 7.45 (2H,d,J=7Hz), 7.90

(2E,d,J=8Ez).

The entire amount of ethyl hexenate derivative (22.3 g) was dissolved in an ethanol-acetic acid mixture (20:1, 200 ml), after which 5% Pd-C (5.0 g) was added, and then stirred extensively for 115 hours. After filtering the Pd-C using celite and evaporating the solvent under reduced pressure, the residue was subjected to evaporation under reduced pressure to give the final compound (15.66 g) as a colorless oil.

K.pt. 162-165°C/0.3mmHg

IR (Neat): 2980, 2940, 1735, 1710, 1607, 848 cm-<1>.

<i>H-NMR (CDCl 3 ) δ: 1.20 (3E,t,J=7Hz), 1.23

(3H,d,J=6Hz), 1.30-1.80 (4H,m), 1.58 (9H,s), 2.24 (2E,brt,J=6Ez), 2.77 (1H ,dq,J=6Hz, 6Hz), 4.08 (2H,q,J = 7Ez ), 7.20 (2H,d,J=8Hz), 7.90 (2H,d,J=8Hz).

Reference example 18

Preparation of ethyl 5-[4-(tert-butoxycarbonyl)phenyl]-2-iodohexanate: The compound (6.41 g) provided in Reference Example 17 was subjected to the same reaction as in Reference Example 2 to give the final compound (3.90 g).

IR (Neat): 2980, 2940, 1738, 1715, 1610, 850 cm-<1.>

<i>H-NMR (CDCl3) S: 1.23 (3H,t,J=7Hz), 1.23

(2H,d,J=7Hz), 1.40-1.95 (4H,m), 1.60 (9H,s), 2.75 (lE.dq,J=6Hz,6Hz), 4.15 (2H,q,J=7Ez), 4.18

(lE,t,J=7Hz), 7.20 (2E,d,J=8Ez), 7.90 (2E,d,J=8Ez).

Reference example 19

Preparation of ethyl 5-[4-(tert-butoxycarbonyl)-phenyl]-2-(dicyanomethyl)-hexanoate: The compound (3.90 g) provided in Reference Example 18 was subjected to the same reaction as that in Reference Example 3 to give the final compound 83.19 g).

IR (Neat): 2980, 2930, 2250, 1735, 1710, 1605, 857 cm-<1>.

<1->H-NMR (CDCl3) S: 1.26 (1.5H, t, J=7Hz ), 1.26

(3E,d,J=7Hz), 1.27 (1.5H,t,J=7Hz), 1.35-1.80 (4H,m),

1.58 (9E,s), 2.50-3.08 (2E,m), 4.00

(lH,dd,J=8Hz,4Ez), 4.22 (1E,q,J=7Ez), 4.23

(lE.q,J=7Ez), 7.18 (2E,d,J=8Ez), 7.92 (2E,d,J=8Ez).

Working example 1

Preparation of tert-butyl 4-[3-(2,4-diamino-6-hydroxy-7E-pyrrolo[2,3-d]pyrimidin-5-yl)propyl]benzoate: To a solution of potassium tert-butoxide (2 .35 g) and guanidine hydrochloride (1.07 g) in tert-butyl alcohol (10 ml), a solution of the compound (3.33 g) provided in Reference Example 3 in tert-butyl alcohol (30 ml) under an argon atmosphere, and heated at reflux for 20 hours. To the reaction mixture was added additional potassium tert-butoxide (434 mg) and guanidine hydrochloride (370 mg) and the mixture was refluxed for an additional 8 hours. The reaction mixture was cooled, added to 1 N potassium hydrogen sulfate in water (about 10 mL) and adjusted to pE 9. After extraction with a tetrahydrofuran-chloroform mixture, the solvent was evaporated under reduced pressure and the resulting residue was purified by column chromatography (support; silica gel, 100 g, eluent; dichloromethane: ethanol = 15:1 -> dichloromethane after mixing with concentrated aqueous ammonia in a separatory funnel: ethanol = 15:1), to give the final compound (1.90 g).

IR (KBr): 3430, 3360, 1710, 1627, 1583, 1432 cm-<1>.

1-E-NMR (CDCl3-Me2SO-d6) S: 1.15-1.73 (2E,m),

1.55 (9E,s), 1.73-2.10 (2E,m), 2.61 (2E,t,J=7Ez),

3.35 (lE.t,J=6Ez), 5.40 (2E,brs), 5.51 (2E,brs),

6.30 (lE.brs), 7.12 (2E,d,J=8Ez), 7.29 (2E,d,J=8Ez ).

Work example 2

Preparation of tert-butyl 4-[3-(2,4-diamino-7E-pyrrolo[2,3-d]pyrimidin-5-yl)propyl]benzoate: To a solution of the compound (575 mg) provided in working example 1 in tetrahydrofuran (6 ml) was a solution of borane-tetrahydrofuran complex (7.5 mmol) in tetrahydrofuran (7.5 ml) at 0°C and stirred for 4.5 hours. To the reaction mixture was added acetic acid-methanol (1:1, 6 ml) and the mixture was stirred at room temperature for 15 hours. The solvent was evaporated under reduced pressure and the residue was purified by column chromatography (support; silica gel, 30 g, eluent; dichloromethane: ethanol = 100:6 -» 100:7 -» 100:8 10:1 -» 8:1), for to give the final compound (263 mg).

IR (KBr): 3335, 3180, 2975, 2935, 1710, 1607,

1207, 1163, 1110 cm-<1>.

<1>H-NMR (Me2S0-d6) S: 1.54 (9H,s), 1.77-1.90 (2H,m), 2.68(2H,t,J=8Hz), 2, 72 (2H,t,J=8Hz), 5.54 (2H,brs),

6.11 (2H,brs), 6.45 (lH,s), 7.33 (2H,d,J=8Hz),

7.82 (2H,d,J=8Hz), 10.51 (1H,s).

Work example 3

Preparation of diethyl N-[4-[3-(2,4-diamino-7H-pyrrolo[2,3-d]pyrimidin-5-yl)propyl]benzoyl]-L-glutamate: To the compound (381 mg) provided in working example 2, trifluoroacetic acid (3 ml) was added and the mixture was stirred at room temperature for 3 hours. Trifluoroacetic acid was evaporated under reduced pressure.

Work example 4

Preparation of tert-butyl 4[3-(2,4-diamino-6-oxo-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)propyl]benzoate: To a solution of potassium tert-butoxide (2.35 g) and guanidine hydrochloride (1.07 g) in tert-butyl alcohol (10 ml), a solution of the compound (3.33 g) provided in Reference Example 16 in tert-butyl alcohol (30 ml) was added under an argon atmosphere, and heated under reflux for 20 hours. To the reaction mixture was added additional potassium tert-butoxide (434 mg) and guanidine hydrochloride (370 mg), and heated at reflux for an additional 8 hours. The reaction mixture was cooled, added to 1N potassium hydrogen sulfate in water (about 10 mL), adjusted to pB 9. After extraction with a tetrahydrofuran-chloroform mixture, the solvent was evaporated under reduced pressure, and the resulting residue was purified by column chromatography (support; silica gel, 100 g, dichloromethane-ethanol; 15:1 dichloromethane after mixing with concentrated aqueous ammonia in a separatory funnel-ethanol, 15:1), to give the final compound (1.90 g).

IR (KBr): 3430, 3360, 1710, 1627, 1583, 1432 cm-<1>.

1-H-NMR (CDCl3/Me2SO-d6) S: 1.15-1.73 (2H,m), 1.55

(9H,s), 1.73-2.10 (2H,m), 2.61 (2H,t,J=7Hz ), 3.35 (lH,t,J=6Hz), 5.40 (2H ,brs), 5.51 (2H,brs), 6.30 (IH.brs), 7.12 (2H,d,J=8Hz), 7.29 (2H,d,J=8Hz).

Work example 5

Preparation of tert-butyl 4-[3-(2,4-diamino-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)propyl]benzoate: To a solution of the compound (430 mg) provided in working example 9 in tetrahydrofuran (10 ml), a solution of the borane-tetrahydrofuran complex (16.8 mmol) in tetrahydrofuran (10 ml) was added and the mixture was refluxed for 4 hours. After cooling, the reaction mixture was added to ice water and thoroughly stirred at pH 2 (adjusted by addition of 1N hydrochloric acid) for 3 minutes at pH 10.5 (adjusted by addition of 2 N potassium carbonate in water) for 5 minutes. The reaction mixture was extracted with tetrahydrofuran-chloroform mixture. The solvent was evaporated under reduced pressure, and the residue was purified by column chromatography (support; silica gel, 20 g, dichloromethane-ethanol, 30:1-»15:1 dichloromethane after mixing with concentrated aqueous ammonia in a separatory funnel-ethanol, 20:1 ), to give the final compound (114 mg).

IR (KBr): 3375, 3325, 3190, 2970, 2930, 1712,

1603 cm-<1>.

1 H-NMR (CDCl 3 /Me 2 SO-d 6 ) S: 1.45-2.15 (4H,m), 1.57

(9H,s), 2.65 (2H,t,J=7Hz), 3.00-3.28 (2H,m),

3.44-3.70 (lH,m), 4.85 (2H,brs), 4.90 (2H,brs), 5.30 (lE.brs), 7.19 (2E,d,J= 8Ez), 7.80 (2H,d,J=8Hz).

Work example 6

Preparation of diethyl N-[4-[3-(2-amino-4-hydroxy-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)propyl]benzoyl]-L-glutamate: The compound (1 .47 g) provided in Reference Example 10 was suspended in tetrahydrofuran (60 ml), whereupon 0.1 N sodium hydroxide in water (120 ml) was added, and the mixture was stirred at room temperature for 21 hours. The solution was then neutralized with 0.1 N hydrochloric acid (60 ml) and concentrated to dryness under reduced pressure.

Work example 7

Preparation of diethyl N-[4-[3-(2,4-diamino-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)propyl]benzoyl]-L-glutamate: The compound (94 mg) provided in working example 10 was dissolved in 1 ml of trifluoroacetic acid and stirred at room temperature for 3 hours. The solvent was evaporated under reduced pressure.

Work example 8

Preparation of tert-butyl 4-[3-(2,4-diamino-6-hydroxy-7E-pyrrolo[2,3-d]pyrimidin-5-yl)-1-methylpropyl]benzoate: The compound (3.18 g ) provided in Reference Example 19 was subjected to the same reaction as that in Working Example 1, to give the final compound (2.61 g).

IR (KBr): 3360, 3235, 2975, 2700, 1715, 1625, 1584,

1438, 1290, 1163, 1118, 848 cm-<1>.

^E-NMR (Me2S0-d6) S: 1.14 (3H,d,J=7Hz), 1.20-1.50

(2E,m), 1.54 (9E,s), 1.55-1.80 (lH,m), 1.80-2.05

(lH,m), 2.60-2.78 (lE.m), 3.20-3.30 (lE,m), 5.86 (2E,brs), 5.96 (2E,brs), 7.25 (2E,d,J=8Hz), 7.81

(2H,d,J=8Hz), 10.42 (1H,s).

Work example 9

Preparation of tert-butyl 4-[3-(2,4-diamino-6,7-dihydro-5E-pyrrolo[2,3-d]pyrimidin-5-yl)-1-methylpropyl]-benzoate (A) and tert-butyl 4-[3-(2,4-diamino-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-1-methyl-propyl]benzoate (B): For a suspension of the compound (2.00 g) provided in working example 17 in tetrahydrofuran (25 ml) became a solution of bolane-tetrahydrofuran complex (40.3 mmol) in tetrahydrofuran (40.3 ml). After stirring for 10 minutes, the mixture was cooled to room temperature and further stirred for 5 hours. To the reaction mixture was added an acetic acid-methanol mixture (1:1, 40 ml) and the mixture was stirred for 18 hours at room temperature. After evaporation of the solvent under reduced pressure, the resulting residue was purified by column chromatography (carrier: silica gel, 100 g, eluent; dichloromethane-ethanol=20 : 1-*25 :2 , then dichloromethane-ethanol containing ammonia (6$) ), to give the final compound (A) (579 mg) and (B) (1.214 g). (A) IR (KBr): 3350, 3200, 2980, 2940m, 1714, 1650, 1608, 1290, 1163, 1115, 848 cm-<1>.

<1->H-NMR (CDCl 3 ) S: 1.31 (3E,d, J=7Ez ), 1.60 (9H,s),

1.94 (2H,dt,J=8Hz), 2.40-2.60 (2H,m), 2.85 (lH,tq,J=7Hz,7Hz), 4.50-5.50 (4H ,br), 6.46 (lH,s),

7.27 (2H,d,J=8Hz), 7.96 (2H,d,J=8Hz), 9.20 (1H.brs).

(B) IR (KBr): 3340, 3195, 2980, 2935, 1715, 1607, 1430,

1295, 1163, 1115, 847 cm-<1>.

1-E-NMR (Me2S0-d6) S: 1.18 (2.25H,d,J=7Hz ), 1.19 (0.75H,d,J=7Hz), 1.23-1.42 ( 2H,m), 1.45-1.65 (2H,m),

1.54 (9E,s,), 2.64-2.70 (lE,m), 2.90-3.08 (2E,m), 3.30-3.50 (lE,m), 5 .43 (4E,s), 5.95 (0.25E,s), 6.00 (0.75E,s), 7.32 (2E,d,J=8Hz), 7.82 (2E,d ,J=8Ez).

Claims (1)

Intermediate with formula: characterized in that the ring A is a pyrrole or pyrroline ring, X is an amino group or a hydroxyl group, Y is a hydrogen atom or a hydroxyl group, B is a straight-chain or branched alkylene with 2-4 C atoms and R<3> is a hydrogen atom or a linear or branched alkyl of 1-4 carbon atoms, or a salt thereof.