Pyrrolopyrimidine derivatives, their production and use

A compound of the formula ##STR1## wherein the ring .circle.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.sup.1 and --COOR.sup.2 are independently carboxyl groups which may be esterified and n is an integer of 2 to 4, and R may be different in each of the n repeating units, and salts thereof have excellent antitumor effects, and can be used as antitumor agents in mammals.

This invention relates to the novel pyrrolopyrimidine derivatives which are 
useful as anti-tumor agents, the production and utilization thereof. 
Folic acid is a carrier of a C1 unit in a living body, derived from formic 
acid or formaldehyde, acting as a coenzyme in various enzymatic reactions 
such as those in biosynthesis of nucleic acid, in metabolism of amino 
acids and peptides and in generation of methane. Particularly in 
biosynthesis of nucleic acid, folic acid is essential for formylation in 
the two pathways, i.e. the purine synthetic pathway and the thymidine 
synthetic pathway. Usually folic acid is required to be transformed into 
its activated coenzyme form by reduction in two steps before it becomes 
biologically active. Amethopterin (methotrexate: MTX) and the related 
compounds are known to inhibit the reduction from dihydrofolic acid into 
tetrahydrofolic acid by coupling strongly with the dominant enzyme in the 
second step (dihydrofolic acid reductase). These drugs have been developed 
as antitumor drugs because they may disturb the DNA synthesis and 
consequently cause cell death, and are currently regarded of major 
clinical important. On the other hand, a novel 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 of which consists in 
inhibition of glycinamide ribonucleotide transformylase required in the 
initial stage of purine biosynthesis [Journal of Medicinal Chemistry, 28, 
914 (1985)]. 
Various studies are now being conducted on therapy for cancer, and what is 
expected strongly is the development of drugs which are more effective and 
have toxicities highly specific to cancer cells based on some new 
mechanism. The antitumor agent MTX the action mechanism of which consists 
in antagonism against folic acid, is clinically used widely, though the 
therapeutic effect is still unsatisfactory because it has relatively 
strong toxicity with little effect on solid cancer. 
As the result of the inventors, researches under the circumstances 
described above, they have found out that novel pyrrolopyrimidine 
derivatives have toxicities highly specific to tumor cells and excellent 
antitumor effects, and completed this invention. 
This invention relates to 
(1) A compound of the formula (I) 
##STR2## 
wherein the ring .circle.A 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.sup.1 and --COOR.sup.2 are 
independently carboxyl groups which may be esterified and n is an integer 
of 2 to 4, and R may be different in each of the n repeating units, and 
salts thereof, (2) A method for production of the compounds (I) or salts 
thereof characterized in that a compound of the formula (II) 
##STR3## 
wherein the ring .circle.A 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, and n is an integer of 2 to 4, and R 
may be different in each of the n repeating units, a reactive derivative 
at the carboxyl group, or a salt thereof, and a compound of the formula 
(III) 
##STR4## 
wherein --COOR.sup.1 and --COOR.sup.2 are independently carboxyl groups 
which may be esterified, or a salt thereof, are allowed to react. 
(3) A compound of the formula (IV) 
##STR5## 
wherein the ring .circle.A a pyrrole or pyrroline ring, X is an amino 
group or a group, Y is a hydrogen atom, an amino group or a group, R is a 
hydrogen atom, a fluorine atom, an alkyl group, an alkenyl group or an 
alkynyl group, --COOR.sup.3 is a carboxyl group which may be esterified 
and n is an integer of 2 to 4, and R may be different in each of the n 
repeating units, and salts thereof. 
(4) Anti-tumor agents containing the compounds (I) or salts thereof. 
When X or Y in the formulas described above is a hydroxyl group, each of 
the compounds (I), (II) and (IV) may exist as an equilibrium mixture of 
the respective tautomers. The following partial structural formulas show 
the sites of the structure which are subject to tautomerism, and the 
equilibrium between the tautomers is illustrated in the following. 
##STR6## 
For the convenience of description, only the hydroxyl forms and the 
corresponding names are described throughout this specification, but the 
corresponding oxo forms are always included. 
There may be two or more asymmetric centers in the compounds (I) of this 
invention, and the absolute configuration at all of the asymmetric carbon 
atoms may be the S, R or S--R mixed form, except that the absolute 
configuration at the asymmetric carbon atom in the side chain derived from 
glutamic acid is always S(L). Therefore the compounds (I) may have two or 
more diastereomers which, if necessary, can easily be separated from each 
other by a routine method for separation and purification. All of the 
diastereomers which can be separated by such a method are included in this 
invention. 
Alkyl groups represented by R in the formulas described above include alkyl 
groups having 1 to 3 carbon atom(s) each (e.g. methyl, ethyl, propyl, 
isopropyl groups). Alkenyl groups represented by R in the formulas 
described above include alkenyl groups having 2 to 3 carbon atom(s) each 
(e.g. vinyl, 1-methylvinyl, 1-propenyl, allyl, allenyl groups. Alkynyl 
groups represented by R in the formulas described above include alkynyl 
groups having 2 to 3 carbon atom(s) each (e.g. ethynyl, 1-propynyl, 
propargyl groups). Carboxyl groups in the carboxyl groups which may be 
esterified, represented by --COOR.sup.1, --COOR.sup.2 and --COOR.sup.3 
include carboxyl groups which may be esterified by alkyl groups having 1 
to 5 carbon atom(s) each, benzyl groups which may be substituted or phenyl 
groups which may be substituted. 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, The benzyl groups 
which may be substituted include benzyl, nitrobenzyl, methoxybenzyl groups 
and so on. The phenyl groups which may be substituted include phenyl, 
nitrophenyl, methoxyphenyl groups and so on. 
In the following the method for production of the compounds (I) of this 
invention is explained. 
The compounds (I) can be obtained by acylation of glutamic acid derivatives 
shown by the formula (III) with carboxylic acids shown by the formula (II) 
or reactive derivatives thereof. The acylation may be performed, for 
example, by acylation of the compound (III) with the compound (II) in the 
presence of carbodiimide, dephenylphosphoryl azide or diethyl phosphoro 
cyanidate. Generally about 1 to 20 mole equivalent, preferably 1 to 5 mole 
equivalent of the compound (III) relative to the compound (II) is used. 
Generally about 1 to 25 mole equivalent, preferably about 1 to 5 mole 
equivalent of a carbodiimide relative to the compound (II) is used. As the 
carbodiimide, dicyclohexylcarbodiimide is preferable for practical use, 
but other carbodiimides such as diphenylcarbodiimide, 
di-o-tolylcarbodiimide, di-p-tolylcarbodiimide, di-tert-butylcarbodiimide, 
1-cyclohexyl-3-(2-morpholinoehtyl)carbodiimide, 
1-cyclohexyl-3-(4-diethylaminocyclohexyl)carbodiimide, 
1-ethyl-3-(2-diethylaminopropyl)carbodiimide and 
1-ethyl-3-(3-diethylaminopropyl)carbodiimide may be used. The acylation is 
preferably performed in the presence of a suitable solvent, and such 
solvents include water, alcohols (e.g. methanol, ethanol, etc.), ethers 
(e.g. dimethyl ether, diethyl ether, tetrahydrofuran, dioxane, monoglyme, 
diglyme, etc.), nitriles (e.g. acetonitrile, etc.), esters (e.g. ethyl 
acetate, etc.), halogenated hydrocarbons (e.g. dichloromethane, 
chloroform, carbon tetrachloride, etc.), aromatic hydrocarbons (e.g. 
benzene, toluene, xylene, etc.), acetone, nitromethane, pyridine, 
dimethylsulfoxide, dimethylformamide, hexamethylphospholamide, sulfolane, 
and the suitable mixtures of two or more of these solvents. The reaction 
is allowed to proceed generally at a pH ranging from 2 to 14, preferably 
at a pH ranging from about 6 to 9, at a temperature ranging from about 
-10.degree. C. to the boiling point of the solvent used (up to about 
100.degree. C.), preferably at a temperature ranging from about 0.degree. 
to 50.degree. C., for about 1 to 100 hours. The pH of the reaction mixture 
is adjusted, if necessary, by addition of an acid (e.g. hydrochloric acid, 
sulfuric acid, phosphoric acid, nitric acid, acetic acid, etc.), a base 
(e.g. sodium alcoholate such as sodium methylate and sodium ethylate, 
hydroxides of alkali metals or of alkali earth metals such as sodium 
hydroxide, potassium hydroxide, lithium hydroxide, barium hydroxide, 
carbonates or bicarbonates of alkali metals or of alkali earth metals such 
as sodium carbonate, potassium carbonate, barium carbonate, calcium 
carbonate and sodium biccarbonate, amines such as trimethylamine, 
triethylamine, triethanolamine and pyridine), or a buffer (e.g. phosphate 
buffer, borate buffer, acetate buffer, etc.). The reaction can proceed 
more advantageously in the presence of a catalyst which promotes 
acylation. Such catylysts include base catalysts and acid catalysts. The 
base catalysts include tertiary amines (e.g. aliphatic tertiary amines 
such as triethylamine; aromatic tertiary amines such as pyridine, 
.alpha.-, .beta.- or .gamma.-picoline, 2,6-lutidine, 
4-dimethylaminopyridine, 4-(1-pyrrolidinyl)pyridine, dimethylaniline and 
diethylaniline), and such acid catalysts include Lewis acids [e.g. 
anhydrous zinc chloride, anhydrous aluminum i5 chloride (AlCl.sub.3), 
anhydrous ferric chloride, titanium tetrachloride (TiCl.sub.4), tin 
tetrachloride (SnCl.sub.4), antimony pentachloride, cobalt chloride, 
cupric chloride, boron trifluoride ethyl ether complex, etc.]. Among the 
catalysts described above, 4-dimethylaminopyridine or 
4-(1-pyrrolidinyl)pyridine is preferable in many cases. The suitable 
amount of the catalyst is such that is enough to promote the acylation, 
being generally about 0.01 to 10 mole equivalent, preferably about 0.1 to 
1 mole equivalent relative to the compound (II). The reactive derivatives 
of carboxylic acids obtained by the reaction at the carboxyl group, used 
for the acylation include acid halides (e.g. fluoride, chloride, bromide, 
iodide), acid anhydrides (e.g. iodoacetic acid anhydride, isobutyric acid 
anhydride), mixed acid anhydrides with monoalkylcarbonic acid esters (e.g. 
mono-methylcarbonic acid ester, monoethylcarbonic acid ester, 
monopropylcarbonic ester, mono-iso-propylcarbonic acid ester, 
monobutylcarbonic acid ester, mono-iso-butylcarbonic acid ester, 
mono-sec-butylcarbonic acid ester, mono-tert-butylcarbonic acid ester), 
active esters (e.g. cyanomethyl ester, carboethoxymethyl ester, 
methoxymethyl ester, phenyl ester, o-nitrophenyl ester, p-nitrophenyl 
ester, p-carbomethoxyphenyl ester, p-cyanophenyl ester, thiophenyl ester), 
acid azides, mixed acid anhydrides with phosphoric acid diesters (e.g. 
dimethyl phosphate, diethyl phosphate, dibenzylphosphate, 
diphenylphosphate), and mixed acid anhydrides with phosphorous acid 
diesters (e.g. dimethyl phosphite, diethyl phosphite, dibenzyl phosphite, 
diphenyl phosphite), of the carboxylic acid (II). For acylation with such 
a reactive derivative, the solvent, the catalyst and the reaction 
temperature are the same as for acylation in the presence of the 
carbodiimide described above. 
For production of the compound (I-1) in which --COOR.sup.1 and --COOR.sup.2 
in the formula of the compound (I) are carboxyl groups, it is desirable 
that the compound in which --COOR.sup.1 and --COOR.sup.2 in the formula of 
the compound (III) are esterified carboxyl groups is allowed to react with 
the compound (II) followed by deesterification by per se known degradation 
or catalytic reduction. Such degradation can be performed by hydrolysis 
under basic conditions (method A), hydrolysis under acidic conditions 
(method B-1) or hydrolysis under acidic nonaqueous conditions (method 
B-2). Bases used in the method A include metal alkoxides such as sodium 
methoxide, sodium ethoxide, sodium butoxide and potassium butoxide, metal 
hydroxides such as sodium hydroxide, potassium hydroxide, lithium 
hydroxide and barium hydroxide, and amines such as ammonia, triethylamine 
and pyridine. Acids used in the method B-1 include mineral acids such as 
hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid and 
phosphoric acid, and organic acids such as trifluoroacetic acid, 
trichloroacetic acid, methanesulfonic acid, benzenesulfonic acid, 
p-toluenesulfonic acid and camphorsulfonic acid. Catalysts used in the 
method B-2 include mineral acids such as hydrogen chloride, hydrogen 
bromide, perchloric acid, sulfuric acid, nitric acid and phosphoric acid, 
organic acids such as trifluoroacetic acid, trichloroacetic acid, 
methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid and 
camphorsulfonic acid, and Lewis acids such as anhydrous zinc chloride, 
anhydrous aluminum chloride (AlCl.sub.3), anhydrous ferric chloride, 
titanium tetrachloride (TiCl.sub.4), tin tetrachloride (SnCl.sub.4), 
antimony pentachloride, cobalt chloride, cupric chloride and boron 
trifuluoride ethyl ether complex. Degradation is performed in a suitable 
solvent at a temperature ranging from 0.degree. C. to the boiling point of 
the solvent, preferably at 10.degree. to 80.degree. C., for 30 minutes to 
2 days. The solvent used for the reaction by the method A or by the method 
B-1 may be water, methanol, ethanol, propanol, butanol, ethyleneglycol, 
methoxyethanol, ethoxyethanol, tetrahydrofuran, dioxane, monoglyme, 
diglyme, pyridine, dimethylformamide, dimethylsulfoxide or sulfolane, or a 
suitable mixture of two or more of these solvents; the solvent used for 
the reaction by the method B-2 may be ethyl acetate, dimetyl ether, 
diethyl ether, tetrahydrofuan, dioxane, monoglyme, diglyme, 
dichloromethane, chloroform, carbon tetrachloride, acetonitrile, benzene, 
toluene, xylene, nitromethane or pyridine, or a suitable mixture of two or 
more of these solvents. The catalytic reduction (method C) is performed in 
a suitable solvent at a temperature ranging from about -40.degree. C. to 
the boiling point of the solvent used, preferably at about 0.degree. to 
50.degree. C.. The solvents used include water, alcohols (e.g. methanol, 
ethanol, propanol, iso-propanol, butylalcohol, sec-butylalcohol, 
tert-butylalcohol, ethyleneglycol, methoxyethanol, ethoxyethanol), acetic 
acid esters (e.g. methyl acetate, ethyl acetate), ethers (e.g. dimethyl 
ether, diethyl ether, tetrahydrofuran, dioxane, monoglyme, diglyme, 
aromatic hydrocarbons (e.g. benzene, toluene, xylene), pyridine, 
dimethylformamide and suitable mixtures of two or more of these solvents. 
Catalysts for the catalytic reaction include palladium, platinum, rhodium 
and Raney nickel. Addition of a trace amount of acetic acid, 
trifluoroacetic acid, hydrochloric acid or sulfuric acid can allow the 
reaction to proceed advantageously. 
The method for production of the compound (I-1) is selected according to 
the nature of --COOR.sup.1 and --COOR.sup.2 ; when --COOR.sup.1 and 
--COOR.sup.2 are carboxyl groups esterified with methyl, ethyl, propyl, 
butyl, sec-butyl, phenyl or substituted phenyl group, the method A or the 
method B-1 is applied advantageously; when --COOR.sup.1 and --COOR.sup.2 
are carboxyl groups esterified with iso-propyl or tert-butyl group, the 
method B-2 is applied advantageously; and when --COOR.sup.1 and 
--COOR.sup.2 are carboxyl groups esterified with benzyl or a substituted 
benzyl group, the method B-1 or the method C is applied advantageously. 
When --COOR.sup.1 and --COOR.sup.2 are different from each other, the 
methods A, B-1, B-2 and C may be combined appropriately. 
In the following the method for production of the starting compound (II) is 
explained. 
The compound (II) wherein the ring .circle.A a pyrrole ring, can be 
produced, for example, by the following processes. 
##STR7## 
In the reaction formulas described above, X, Y and R.sup.3 are the same as 
described before; R.sup.a, R.sup.b and R.sup.c are independently a 
hydrogen atom, a fluorine atom or an alkyl group (the same as those 
represented by R described before); R.sup.4 is a cyano group or an 
esterified carboxyl group represented by the formula --COOR.sup.5 ; A is a 
hydrogen atom or a halogen atom (e.g. fluorine atom, chlorine atom, 
bromine atom, iodine atom); B is a halogen atom (e.g. chlorine atom, 
bromine atom, iodine atom) or an eliminable group which may be easily 
derived from hydroxy group (e.g. methanesulfonyloxy group, 
benzenesulfonyloxy group, p-toluenesulfonyloxy group, 
trifluoromethanesulfonyloxy group); and m is 0, 1 or 2. R.sup.5 in the 
esterified carboxyl group represented by the formula --COOR.sup.5 is 
exemplified by an alkyl group having 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-methoxybenzyl, etc.). 
The compound (V) may be dehydrogenated on the possible position between the 
two adjacent carbons and form an unsaturated bond. 
In the following the reaction processes described above are explained in 
detail. 
Process 1 
The compound (V) and the compound (VI) are subjected to condensation and 
the resulting product is subjected to reduction to give the compound 
(VII). 
For the condensation, a known reaction (e.g. aldol reaction, Reformatsky 
reaction, Wittig reaction, etc.) is employable, and for the reduction, 
usually a catalytic reduction under hydrogen atmosphere in the presence of 
a catalyst (e.g. nickel, palladium, platinum, rhodium) is advantageously 
employed. 
In the condensation by aldol reaction, the employable base catalysts 
include metal hyrdoxides 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, 
.alpha.-, .beta.- or .gamma.-picoline, 2,6-lutidine, 
4-dimethylaminopyridine, 4-(1-pyrrolidinyl)pyridine, dimethylaniline and 
diethylaniline; the employable 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. The condensation can be 
conducted according to the known method [Ei-Ichi Negishi, Organometallics 
in Organic Synthesis, vol. 1, John Wiley & Sons, New York, Chichester, 
Brisbane, Toronto (1980)]which comprises converting a ketone form into the 
silylenolether form which is then subjected to condensation with an 
aldehyde or an equivalent in the presence of a Lewis acid [e.g. anhydrous 
zinc chloride, anhydrous aluminum chloride (AlCl.sub.3), anhydrous ferric 
chloride, titanium tetrachloride (TiCl.sub.4), tin tetrachloride 
(SnCl.sub.4), antimony pentachloride, cobalt chloride, cupric chloride, 
boron trifluoride ethyl ether complex, etc.], or converting a ketone form 
into the enolate by treating the ketone form with a metal triflate (e.g. 
dialkyl boron or tin (II) triflate) in the presence of amines (e.g. 
triethylamine, pyridine, .alpha.-, .beta.- or .gamma.-picoline, 
2,6-lutidine, 4-dimethylaminopyridine, 4-(1-pyrrolidinyl)pyridine, 
dimethylaniline, diethylaniline) followed by subjecting the enolate to 
condensation with an aldehyde or an equivalent. The condensation is 
conducted in a suitable solvent at a temperature ranging from -100.degree. 
C. to the boiling point of 
the solvent, preferably ranging from -78.degree. to 100.degree. C., for 1 
minute to 3 days. Solvents employable for 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), ethers (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 the suitable 
mixtures thereof. In the condensation by Wittig reaction, the employable 
reagents 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, .alpha.-, .beta.- or .gamma.-picoline, 2,6-lutidine, 
4-dimethylaminopyridine, 4-(1-pyrrolidinyl)pyridine, dimethylaniline and 
diethylaniline. The reaction is conducted in a suitable solvent at a 
temperature ranging from -20.degree. C. to the boiling point of the 
solvent used, preferably ranging from 0.degree. to 150.degree. C., for 1 
minute to 10 days. The solvents employable for the reaction include liquid 
ammonia, alcohols (e.g. methanol, ethanol, propanol, isopropanol, butyl 
alcohol, sec-butyl alcohol, tert-butyl alcohol, ethylene glycol, 
mehoxyethanol, ethoxyethanol), ethers (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, hexamethylphospholam:ide, 
sulfolane and the suitable mixtures thereof. 
The condensation can also be conducted by using a Reformatsky reaction. The 
reagents employable for the Reformatsky reaction include zinc, magnesium, 
aluminum and tin, and the reaction is conducted in a suitable solvent at a 
temperature ranging from -20.degree. C. to the boiling point of the 
solvent used, preferably ranging from 0.degree. to 150.degree. C., for 30 
minutes to 3 days. The solvents employable for the reaction include ethers 
(e.g. dimethyl ether, diethyl ether, tetrahydrofuran, dioxane, monoglyme, 
diglyme), aliphatic hydrocarbons (e.g. pentane, hexane, heptane), aromatic 
hydrocarbons (e.g. benzene, toluene, xylene) and the suitable mixtures 
thereof. 
The reaction conditions for the catalytic reduction are the same as those 
for the deesterification at the --COOR.sup.1 and --COOR.sup.2 of the 
compound (III) (method C). 
The starting materials (V) and (VI) can be obtained easily according to the 
known methods described in the literature. [B. Neises et al., Angew. Chem. 
Int. Ed. Engl., 17, 522 (1978)]. 
Process 2 
This is the process whereby an eliminable functional group B is introduced 
into the active methylene (the .alpha.-position of the carbonic acid 
ester) of the compound (VII): it can be conducted easily by using known 
reagents according to a per se known method. 
Process 3 
The compound (VIII) obtained in the Process 2 is subjected to condensation 
with malononitrile or a cyanoacetic acid ester [NC--CH:COOR.sup.5 ; 
R.sup.5 is the same as described above]under a basic condition, to give 
the compound (IX). The employable bases, solvents and reaction conditions 
are in accordance with the known methods. 
Process 4 
The compound (IX), when treated with guanidine, can react at the cyano 
group or the ester residue followed by ring closure to form newly a 
pyrrolopyrimidine ring. Ring closure under a basic condition allows the 
reaction to proceed advantageously. The employable bases include metal 
alkoxides such as sodium methoxide, sodium ethoxide and potassium 
tert-butoxide. The employable solvents for the reaction include methanol, 
ethanol, propanol, tert-butyl alcohol, dimethylsulfoxide and 
hexamethylphospholamide. The reaction temperature ranges from 0.degree. to 
150.degree. C., preferably from 20.degree. to 100.degree. C. The reaction 
time ranges from 1 to 48 hours. 
Process 5 
The compound (IV-1: Y.dbd.NH.sub.2 or OH) obtained in the and --COOR.sup.2 
of the compound (III) (method C). Process 4 can be converted into the 
compound (II-1: Y.dbd.NH.sub.2 or OH) by subjecting the ester residue 
[--COOR.sup.3 ] to the deesterification used in the preparation of the 
compound (I-1). 
Process 6 
The compound (II-1: Y.dbd.OH) obtained in the Process 5 is subjected to 
reduction to give the compound (II-2: Y.dbd.H). The conditions for the 
reduction are per se known, and reduction by a metal hydride (e.g. borane, 
alane or ate complexes thereof) is employed advantageously. 
The Process 5 and the Process 6 may be conducted in the reverse order. 
Namely, in the Process 7 the compound (IV-1: Y.dbd.OH) is subjected to 
reduction similar to that in the Process 6 to give the compound (IV-2: 
Y.dbd.H), which is then subjected to deesterification in the Process 8 in 
a similar manner as in the Process 5 to give the compound (II-2: Y.dbd.H). 
Either the deesterification or the reduction can be selected to be 
conducted in advance to the other according to the nature of the 
substituents in the compound (IV-1: Y.dbd.OH). 
In the above Processes 6 and 8, the mixture containing the compounds (II-2) 
and (II-2') or the compounds (IV-2) and (IV-2') may be separated, or each 
of the compounds (II-2) and (II-2') or each of the compounds (IV-2) and 
(IV-2') is synthesized predominantly by selective reduction. 
Among the compounds (II), those represented by the formula (II-3: X.dbd.OH) 
##STR8## 
wherein R and n mean the same as described before, can be obtained also by 
the following processes. 
##STR9## 
In the Processes described above, R, Ra, Y and n mean the same as described 
before and Z means the formula RCH.sub.2 CO- wherein R means the same as 
described above, the formula 
##STR10## 
wherein L is phenyl, butyl or cyclohexyl, and R and n mean the same as 
described above, or the formula 
##STR11## 
wherein M is ethyl or phenyl, and R and n mean the same as described 
above, It is preferable that Y is hydrogen. 
In the following these Processes are explained. Process 9 
The compound (X) [T. Kondo et al., Chemistry Letters, 19 (1983)]and a 
para-substituted benzoic acid ester derivative (XI) are subjected to 
condensation (aldol reaction, Wittig reaction) followed by catalytic 
reduction under hydrogen atmosphere, to give the compound (XII). For the 
condensation are applicable the reaction conditions, the reaction 
solvents, the reaction temperatures and the reagents used in the Process 
1. For the catalytic reduction under hydrogen atmosphere are applicable 
the conditions used in the deesterification of --COOR.sup.2 and 
--COOR.sup.2 of the compound (III). Process 10 
Treatment of the compound (XII) under acidic conditions can eliminate the 
protection of the isopropyloxymethyl group at the 3-position to give the 
compound (XIII). The conditions, solvents and temperatures used in 
deesterification of --COOR.sup.1 and --COOR.sup.2 of the compound (III) 
(the method B-1 and the method B-2) are employable for the reaction. 
Process 11 
The compound (XIII) obtained in the Process 10 is subjected to 
dehydrogenation by a per se known method, to be easily converted into the 
compound (IV-3: Y.dbd.H). 
Process 12 
The compound (IV-3: Y.dbd.H) obtained in the Process 11 can be converted 
into the compound (II-3) by deesterification. The conditions, solvents and 
temperatures described in detail for the deesterification of --COOR.sup.1 
and --COOR.sup.2 of the compound (III) (the methods A, B-1, B-2 and C) are 
employable for the reaction. The processes 10 to 12 may be conducted in 
any order with the formation of the respective products, and finally the 
desired compound (II-3) is obtained. The order is determined suitably 
according to the nature of the substituents of the compounds (XII), (XIII) 
and (IV-3). The compound (II-3) thus obtained can be converted, if 
necessary, into the compound (II-2) by a known substituent-converting 
reaction on the pyrimidine ring reported in the literature. [Protein 
Nucleic acid Enzyme Extra Issue, Chemical synthesis of nucleic acids, 
Kyoritsu Shuppan (1968)] 
The compounds other than the compound (II-3), wherein X is hydroxyl can be 
also converted into the corresponding compounds wherein X is amino by the 
above-mentioned substituent-converting reaction. 
The reactions, reagents and reaction conditions used in the Processes 1 to 
12 and in the production of the 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 
edition) [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 intermediates of the 
compounds of this invention and the compounds (I) of this invention can be 
isolated from the reaction mixtures by the conventional means for 
separation and purification, such as concentration, extraction with 
solvent, chromatography and recrystallization. The compounds (I), (II) and 
(IV) of this invention may form salts. Such salts are produced by the 
known methods, and exemplified by the salts of pharmaceutically acceptable 
bases or acids and quaternary salts. Salts of bases include salts of 
alkali metals, alkali earth metals, non-toxic metals, ammonium and 
substituted ammonium, such as sodium, potassium, lithium, calcium, 
magnesium, aluminum, zinc, ammonium, trimethylammonium, triethylammonium, 
triethanolammonium, pyridinium and substituted pyridinium. Salts of 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, methyl 
benzensulfonate and methyl p-toluenesulfonate. Also, the compounds (I), 
(II) and (IV) may form zwitterion. 
As the compounds (I) of this invention, the following compounds are 
exemplified: 
Diethyl 
N-[4-[3-(2,4-diamino-7H-pyrrolo[2,3-d]-pyrimidin-5-yl)propyl]benzoyl]-L-gl 
utamate, 
N-[4-[3-(2,4-diamino-7H-pyrrolo[2,3-d]pyrimidin-5-yl)propyl]benzoyl]-L-glut 
amic acid, 
Diethyl 
N-[4-[3-(2,4-diamino-6-hydroxy-7H-pyrrolo-[2,3-d]pyrimidin-5yl)propyl]benz 
oyl]-L-glutamate, 
N-[4-[3-(2,4-diamino-6-hydroxy-7H-pyrrolo[2,3-d]-pyrimidin-5-yl)propyl]benz 
oyl]-L-glutamic acid, 
Diethyl 
N-[4-[3-(2-amino-4-hydroxy-7H-pyrrolo[2,3-d]-pyrimidin-5yl)propyl]benzoyl] 
-L-glutamate, 
N-[4-[3-(2-amino-4-hydroxy-7H-pyrrolo[2,3-d]-pyrimidin-5-yl)propyl]benzoyl] 
-L-glutamic acid, 
Diethyl 
N-[4-[3-(2-amino-4-hydroxy-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)propyl] 
benzoyl]-L-glutamate, 
Diethyl 
N-[4-[2-(2-amino-4-hydroxy-5,6-dihydropyrrolo-[2,3-d]pyrimidin-5-yl)ethyl] 
benzoyl]-L-glutamate, 
Diethyl 
N-[4-[3-(2,4-diamino-5,6-dihydropyrrolo-[2,3-d]pyrimidin-5-yl)propyl]benzo 
yl]-L-glutamate, 
N-[4-[3-(2-amino-4-hydroxy-5,6-dihydropyrrolo[2,3-d]-pyrimidin-5-yl)propyl] 
benzoyl]-L-glutamic acid, 
N-[4-[2-(2-amino-4-hydroxy-5,6-dihydropyrrolo[2,3-d]-pyrimidin-5-yl)ethyl]b 
enzoyl]-L-glutamic acid, 
N-[4-[3-(2,4-diamino-5,6-dihydropyrrolo[2,3-d]-pyrimidin-5-yl)propyl]benzoy 
l]-L-glutamic acid, 
Diethyl 
N-[4-[3-(2,4-diamino-6,7-dihydro-5H-pyrrolo-[2,3-d]pyrimidin-5-yl)-1-methy 
lpropyl]benzoyl]-L-glutamate, 
N-[4-[3-(2,4-diamino-6,7-dihydro-5H-pyrrolo[2,3-d]- 
pyrimidin-5-yl)-1-methylpropyl]benzoyl]-L-glutamic acid, 
Diethyl 
N-[4-[3-(2,4-diamino-7H-pyrrolo[2,3-d]-pyrimidin-5-yl)-1-methylpropyl]benz 
oyl]-L-glutamic acid, 
N-[4-[3-(2,4-diamino-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-1-methylpropyl]benzoy 
l]-L-glutamic acid, 
N-[4-[3-(2,4-diamino-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-methyl-propyl]benzo 
yl]-L-glutamic acid, 
N-[4-[3-( 3-d]pyrimidin-5-yl)-1-ethylpropyl]benzoyl]-L-glutamic acid, 
N-[4-[3-(2,4-diamino-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-1-ethylpropyl]benzoyl 
]-L-glutamic acid, 
N-[4-[3-(2,4-diamino-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-1-vinylpropyl]benzoyl 
]-L-glutamic acid, 
N-[4-[3-(2,4-diamino-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-vinylpropyl]benzoyl 
]-L-glutamic acid, 
N-[4-[3-(2,4-diamino-7H-pyrollo[2,3-d]pyrimidin-5-yl)-1-allylpropyl]benzoyl 
]-L-glutamic acid, 
N-[4-[3-(2,4-diamino-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-allylpropyl]benzoyl 
]-L-glutamic acid, 
N-[4-[3-(2,4-diamino-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-1-(propen-1-yl)propyl 
]benzoyl]-L-glutamic acid, 
N-[4-[3-(2,4-diamino-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-(propen-1-yl)-propy 
l]benzoyl]-L-glutamic acid, 
N-[4-[3-(2,4-diamino-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-1-ethynylpropyl]benzo 
yl]-L-glutamic acid, 
N-[4-[3-(2,4-diamino-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-ethynylpropyl]benzo 
yl]-L-glutamic acid, 
N-[4-[3-(2,4-diamino-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-1-propargylpropyl]ben 
zoyl]-L-glutamic acid, 
N-[4-[3-(2,4-diamino-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-propargylpropyl]ben 
zoyl]-L-glutamic acid, 
N-[4-[3-(2,4-diamino-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-1-(propyn-1-propyl]be 
nzoyl]-L-glutamic acid, 
N-[4-[3-(2,4-diamino-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-1-(propyn-1-yl)propyl 
]benzoyl]-L-glutamic acid, 
N-[4-[3-(2,4-diamino-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-(propyn-1-yl)-propy 
l]benzoyl]-L-glutamic acid, 
N-[4-[3-(2,4-diamino-6,7-dihydro-5H-pyrrolo[2,3-d]-pyrimidin-5-yl)-2-methyl 
propyl]benzoyl]-L-glutamic acid, 
N-[4-[3-(2,4-diamino-6,7-dihydro-5H-pyrrolo[2,3-d]-pyrimidin-5-yl)-2-methyl 
propyl]benzoyl]-L-glutamic acid, 
N-[4-[3-(2,4-diamino-6,7-dihyiro-5H-pyrrolo[2,3-d]-pyrimidin-5-yl)-2-ethylp 
ropyl]benzoyl]-L-glutamic acid, 
N-[4-[3-(2,4-diamino-6,7-dihydro-5H-pyrrolo[2,3-d]-pyrimidin-5-yl)-1-vinylp 
ropyl]benzoyl]-L-glutamic, acid, 
N-[4-[3-(2,4-diamino-6,7-dihydro-5H-pyrrolo[2,3-d]-pyrimidin-5-yl)-2-vinylp 
ropyl]benzoyl]-L-glutamic, acid, 
N-[4-[3-(2,4-diamino-6,7-dihydro-5H-pyrrolo[2,3-d]-pyrimidin-5-yl)-1-allylp 
ropyl]benzoyl]-L-gluta,mic acid, 
N-[4-[3-(2,4-diamino-6,7-dihydro-5H-pyrrolo[2,3-d]-pyrimidin-5-yl)-2-allylp 
ropyl]benzoyl]-L-gluta,mic acid, 
N-[4-[3-(2,4-diamino-6,7-dihydro-5H-pyrrolo[2,3-d]-pyrimidin-5-yl)-1-(prope 
n-1-yl]benzoyl]-L-glutamic acid, 
N-[4-[3-(2,4-diamino-6,7-dihydro-5H-pyrrolo.differential.2,3-d]-pyrimidin-5 
-yl)-2-(propen-1-yl]benzoyl]-L-glutamic acid, 
N-[4-[3-(2,4-diamino-6,7-dihydro-5H-pyrrolo[2,3-d]-pyrimidin-5-yl)-1-ethylp 
ropyl]benzoyl]-L-glutamic acid, 
N-[4-[3-(2,4-diamino-6,7-dihydro-5H-pyrrolo[2,3-d]-pyrimidin-5-yl)-2-ethylp 
ropyl]benzoyl]-L-glutamic, acid, 
N-[4-[3-(2,4-diamino-6,7-dihydro-5H-pyrrolo[2,3-d]-pyrimidin-5-yl)-1-propar 
gylpropyl]benzoyl]-L-glutamic acid, 
N-[4-[3-(2,4-diamino-6,7-dihydro-5H-pyrrolo[2,3-d]-pyrimidin-5-yl)-2-propag 
ylpropyl]benzoyl]-L-glutamic acid, 
N-[4-[3-(2,4-diamino-6,7-dihydro-5H-pyrrolo[2,3-d]-pyrimidin- 
5-yl)-1-(propyn-1-yl)propyl]benzoyl]-L-glutamic acid, 
N-[4-[3-(2,4-diamino-6,7-dihydro-5H-pyrrolo[2,3-d]-pyrimidin-5-yl)-2-(propy 
n-1-yl)propyl]benzoyl]-L-glutamic acid. 
Effects 
The compounds (I) of this invention show excellent antitumor effects in 
mouse tumor cell strains (P388, L1210, L5178Y, B16 melanoma, MethA, Lewis 
Lung Carcinoma, S180 sarcoma, Ehrlich Carcinoma, Colon38) and human tumor 
cell strains (HL60, KB, Lu65), decrease the tumors carried by warm-blooded 
animals [e.g. melanoma, sarcoma, mastocytoma, carcinoma, neoplasia, 
etc.]and prolong the life-span of tumor-carrying warm-blooded animals. 
In the following are described the results indicating the pharmaceutical 
effects of the compounds (I) of this invention. 
The cell growth inhibiting effect (IC.sub.50) of the compounds obtained in 
the Working Examples described below in KB cells was determined by the 
following method. 
Human nasopharyngeal cancer KB cells (1.times.10.sup.4 cells/ml) prepared 
according to a conventional method were inoculated into each well of the 
96-microwell plate (0.1 ml in a well) and subjected to standing culture at 
37.degree. C. under 5% CO.sub.2 for 24 hours. To this was added a solution 
of one of the compounds obtained in the Working Examples in 10% MEM 
(Nissui Pharmaceutical Co. Ltd.), and subjected again to standing culture 
at 37.degree. C. under 5% CO.sub.2 for 72 hours. Then the culture was 
pipetted out, and another 0.1 ml of the solution of MTT (Dojindo 
Laboratories) in 10% MEM (1.0 mg/ml) was added and incubated at 37.degree. 
C. for 4 hours. Then 0.1 ml of the 10% SDS solution (Wako Pure Chemicals) 
was added and incubated at 37.degree. C. for further 24 hours. The 
absorbance at 590 nm was measured and the IC.sub.50 value of the compound 
was defined as the concentration of the compound required to decrease the 
number of cells in the untreated control group by 50%. The results 
obtained are shown in Table 1. Table 1 
TABLE 1 
______________________________________ 
Test compound IC.sub.50 (.mu.g/ml) 
______________________________________ 
Compound of Working Example 4 
0.0003 
Compound of Working Example 6 
0.08 
Compound of Working Example 16 
0.0006 
______________________________________ 
In addition, the following are described the results indicating the 
pharmaceutical effects of the compounds (I) of this invention. 
The cell growth inhibiting effect (IC.sub.50) of the compound obtained in 
the Working Example 14 described below in HL-60 and HEL cells was 
determined by the following method. 
(1) Human Leukemia cells HL-60 (2.times.10.sup.5 cells/ml) were suspended 
in the GIT culture medium (Wako Pure Chemicals) containing the compound of 
this invention and 0.2 ml of the suspension was inoculated into each well 
of the 96-microwell plate. After standing culture at 37.degree. C. under 
5% CO.sub.2 for 68 hours, 1 .mu.Ci of [.sup.3 H]-thymidine (5 Ci/mmol) was 
added and the mixture was incubated for 4 further hours. For measurement 
of the incorporation of thymidine into the cells, the acid-insoluble 
fraction was collected on a glass filter, and the radioactivity of the 
fraction was measured by a liquid scintillation counter. The IC.sub.50 
value of the compound was defined as the concentration of the compound 
required to decrease the radioactivity incorporated into the cells in the 
untreated control group by 50%. 
(2) Human fetal normal lung fibroblasts HEL (1.times.10.sup.4 cells/ml) 
were suspended in the MEM culture medium (Nippon Flow Laboratories) and 
0.1 ml of the suspension was inoculated into each well of the 96-microwell 
plate. After standing culture at 37.degree. C. under 5% CO.sub.2 for 24 
hours, MEM culture medium containing the compound of this invention was 
added and the mixture was incubated for further 72 hours. The medium was 
replaced by the medium containing 1 .mu.g/ml of MTT (Dojindo 
Laboratories), to which was added 10% SDS (Wako Pure Chemicals), and 
incubated overnight. Absorbance at 590 nm was measured by he Multiscan 
(Titertec Co.). The IC.sub.50 value was determined by comparing the 
absorbance in the untreated control group. The results obtained are shown 
in Table 1. 
TABLE 1 
______________________________________ 
Test compound HL-60 (.mu.g/ml) 
HEL (.mu.g/ml) 
______________________________________ 
Compound of Working 
0.04 &gt;20.0 
Example 14 
______________________________________ 
As shown by the above-mentioned compounds (I) are excellent in inhibition 
of cell growth of KB and HL-60, while they do not exert a toxicity against 
HEL. The compounds (I) of this invention and the salts thereof are of 
low-toxicity, having remarkable antitumor effect. Therefore, the 
preparations containing the compound (I) or salts thereof can be employed 
as antitumor agents for the treatment of tumors in warm-blooded animals, 
particularly mammals (e.g. mouse, rat, cat, dog, rabbit, etc.). 
The compounds (I) and salts thereof, when used as antitumor agents, can be 
administered orally and parenterally as they are or in the forms of 
powders, granules, tablets, capsules, suppositories and injections, which 
are prepared according to the conventional methods using pharmaceutically 
acceptable excipients, vehicles, and diluents. The dose varies according 
to the animals, diseases, symptoms, compounds and administration routes; 
for example, the daily dose is about 2.0 to 100 mg of a compound of this 
invention per kg of body weight of a warm-blooded animal described above 
for oral administration, and about 1.0 to 50 mg/kg for parenteral 
administration. Injections may be administered intramuscularly, 
intraperitoneally, subsutaneously or intravenously. 
The preparations are produced by the per se known processes. For the 
above-mentioned oral preparations, for example, tablets are produced by 
suitable combination with a binder (e.g. hydroxypropylcellulose, 
hydroxypropylmethylcellulose, macrogol, etc.), a disintegrator (e.g. 
starch, calcium carboxylmethylcellulose, etc.) and a lubricant (e.g. 
magnesium stearate, talc, etc.). 
As parenteral preparations, for example, injections are produced by 
suitable combination with an agent to provide isotonicity (e.g. glucose, 
D-sorbitol, D-mannitol, sodium chloride, etc.), an antiseptic (e.g. benzyl 
alcohol, chlorobutanol, methyl p-hyrdoxybenzoate, propyl 
p-hydroxybenzoate, etc.) and a buffer (e.g. phosphate buffer, sodium 
acetate buffer, etc.). 
An example process for production of tablets comprises mixing about 1.0 to 
25 mg of the compound of this invention, 100 to 500 mg of lactose, about 
50 to 100 mg of corn starch and about 5 to 20 mg of hydroxypropylcellulose 
for preparation of a tablet by a conventional means, granulating, mixing 
with corn starch and magnesium stearate and tabletting, so that tablets 
each weighing about 100 to 500 mg with the diameter of about 3 to 10 mm 
are obtained. The tablets may be coated with a mixture of acetone and 
ethanol, the mixture containing hydroxypropylmethylcellulose phthalate 
(about 10 to 20 mg per tablet) and castor oil (0.5 to 2 mg) at a 
concentration of about 5 to 10%, to give enteric coated tablets. 
An example process for injectable preparation comprises dissolving about 
2.0 to 50 mg of a sodium salt of the compound of this invention in about 2 
ml of physiological saline for preparation of an ampoule, sealing the 
resultant solution in an ampoule and sterilizing the ampoule at 
110.degree. C. for about 30 minutes or adding about 2 ml of sterile 
distilled water containing about 10 to 40 mg of mannitol or sorbitol into 
the ampoule, freeze-drying and sealing the ampoule. For use of the 
freeze-dried compound for subcutaneous, intravneous or intramuscular 
injection, the ampoule is opened and the content is dissolved in, for 
example, physiological saline so that the concentration of the compound 
may be about 1.0 to 25 mg/ml.

The following Reference Examples and Working Examples will explain the 
present invention more concretely. 
REFERENCE EXAMPLE 1 
Production of methyl 5-[4-(tert-butoxycarbonyl)phenyl]pentanoate: 
Under an atmosphere of argon, potassium (25 g) was added to dried 
tert-butyl alcohol (820 ml), which was refluxed by heating to be dissolved 
completely. The solution was cooled to 20.degree. C., to which ether (300 
ml) was added and then 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 added slowly while the inner temperature was kept at 10.degree. 
C.. After stirring at the same temperature for 2 hours, 1 N potassium 
hydrogen sulfate in water (750 ml) was added with cooling so that the pH 
was adjusted 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 resultant residue 
was dissolved in ethyl acetate (100 ml), to which 5%Pd-C (15 g: Engelhard 
Co. Ltd.) was added and stirred vigorously under hydrogen pressure of 4 
kg/cm.sup.2 at room temperature for 3 hours. The catalyst was filtrated 
off, the solvent was evaporated under reduced pressure, to the residue 
were added dried methanol (200 ml), 4-(N,N-dimethylamino)pyridine (30 mg) 
and dichloromethane (250 ml), and then a solution of 
1,3-dicyclohexylcarbodiim:ide (132 g) in dichloromethane (250 ml) was 
slowly added dropwise at 0.degree. C.. After stirring at room temperature 
for 18 hours, the mixture was cooled to 0.degree. C.; which acetic acid 
(30 ml) was added and the mixture was stirred at 0.degree. C. for 30 
minutes and then at room temperature for 30 minutes. The resultant 
precipitate was filtrated off, the filtrate was concentrated to dryness 
under reduced pressure, to the residue was added ethyl acetate (100 ml) 
and after keeping at 0.degree. C. for 2 hours, the resultant precipitate 
was again filtrated off. The filtrate was concentrated under reduced 
pressure and the residue was purified by column chromatography (carrier; 
silica gel, 100 g, developing solvent; ether: hexane =1:15.fwdarw.1:5), to 
give the object compound (59.7 g). melting point (Bp) 
145.degree.-155.degree. C./0.2-0.3 mmHg 
IR (Neat): 2980, 2950, 1740, 1712, 1605 cm.sup.-1. 
.sup.1 H-NMR (CDCl.sub.3) .delta.: 1.40-1.75 (4H,m), 1.55 (9H,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 2 
Production of methyl 5-[4-(tert-butoxycarbonyl)-phenyl]-2-iodopentanoate: 
Under an atmosphere of argon, to a solution of diisopropylamine (2.48 g) in 
tetrahydrofuran (100 ml) was added a solution of butyllithium (24.5 mmol) 
in hexane (15.3 ml) at 0.degree. C. and stirred for 10 minutes, to this a 
solution of the compound (6.53 g) obtained in the Reference Example 1 in 
tetrahydrofuran (50 ml) was added dropwise at -78.degree. C. over 30 
minutes. After stirring for 30 minutes, a solution of iodine (5.66 g) in 
tetrahydrofuran (30 ml) was added and stirred for further 20 minutes. The 
temperature of the solution was brought up to 0.degree. C. over 30 
minutes, 1 N potassium hydrogen sulfate in water (30 ml) was added 
dropwise, and the solution was extracted with ether after adjustment 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 object compound (4.736 g). 
IR (Neat): 2990, 2905, 1744, 1718, 1612 cm.sup.-1. 
.sup.1 H--NMR (CDCl.sub.3).delta.: 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 (1H,t,J=7Hz), 7.20 
(2H,d,J=8Hz), 7.90 (2H,d,J=8Hz). 
REFERENCE EXAMPLE 3 
Preparation of methyl 
5-[4-(tert-butoxycarbonyl)-phenyl]-2-(dicyanomethyl)pentanoate: 
To a suspension of sodium hydride (1.356 g) in dimethylsulfoxide (8 ml) was 
added a solution of malononitrile (3.37 g) in dimethylsulfoxide (8 ml) 
under cooling with water, and stirred for 15 minutes. To this solution was 
added dropwise a solution of the compound (4.736 g) obtained in the 
Reference Example 2 in dimethylsulfoxide (12 ml) and stirred at room 
temperature for 1 hour, to this 45 ml of 1N potassium hydrogen sulfate in 
water was added at 0.degree. C., followed by extraction 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 (carrier; silica gel, 
200 g, developing solvent; ethyl acetate: hexane=1:5), to give the object 
compound (3.33 g). 
IR (Neat): 2970, 2930, 2252 1740, 1713, 1608 cm.sup.-1 
.sup.1 H--NMR (CDCl.sub.3) .delta.: 1.60-2.05 (4H,m), 1.48 (9H,s), 2.70 
(2H,brt,J=7Hz), 2.90-3.15 (1H,m), 3.82 (3H,s), 4.04 (1H,d,J=7Hz), 7.20 
(2H, d,J=8Hz), 7.92 (2H,d,J=8Hz). 
REFERENCE EXAMPLE 4 
Production of methyl 
4-[3-(2-amino-7-benzyl-3-isopropyloxymethyl-4(3H)-oxopyrrolo[2,3-d]pyrimid 
in-5-yl)-1-oxo-2-propenyl]benzoate: 
2-Amino-7-benzyl-3-isopropyloxymethyl-4(3H)-oxopyrrolo[2,3-d]pyrimidine-5-c 
arbaldehyde (1.7 g) was suspended in a methanol-tetrahydrofuran mixture 
(10:1, 33 ml), to 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 
object compound (2.02 g) as yellow needles. 
IR (KBr): 3480, 3350, 1710, 1680, 1620, 1550, 1375, 1280, 1210, 1110, 1060, 
775. cm.sup.-1. 
.sup.1 H--NMR (CDCL.sub.3) .delta.: 1.23 (6H,d,J=6Hz), 3.93 (3H,s), 
3.80-4.07 (1H,m), 5.15 (2H,s), 5.63 (2H,s), 6.92 (1H,s), 7.10-7.40 (5H, 
m), 7.73 (1H,d,J=15Hz), 8.13 (4H,s), 8.60 (1H,d,J-15Hz). 
REFERENCE EXAMPLE 5 
Production of methyl 
4-[3-(2-amino-3-isopropyloxymethyl-4(3H)-oxo-5,6-dihydropyrrolo-[2,3-d]pyr 
imidin-5-yl)propyl]benzoate: 
The compound (2.01 g) obtained in Reference Example 4 was dissolved in a 
methanol-tetrahydrofuran mixture (3:4, 350 ml), to which 1 N hydrochloric 
acid (8 ml) and 10% Pd-C (4 g, manufactured by Engelhard Co. Ltd.) were 
added, and subjected to catalytic reduction under an atmosphere of 
hydrogen for 48 hours. The catalyst was filtrated off, the filtrate was 
neutralized, the solvent was evaporated off under reduced pressure and the 
residue was isolated and purified by column chromatography on silica gel 
(carrier; 100 g, developing solvent; chloroform containing 2-4% of 
ethanol), to give the object compound (0.68 g) as a colorless powder. 
IR (KBr): 3210, 2980, 1725, 1625, 1580, 1510, 1435, 1275, 1175, 1100, 1060 
cm.sup.-1. 
.sup.1 H--NMR (CDCl.sub.3) .delta.: 1.17(3H,d,J=6Hz), 1.19 (3H,d,J=6Hz), 
1.50-2.13 (4H,m), 2.70 (2H,t,J=7.5Hz), 3.07-3.77 (3H,m), 3.80-4.60 (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 6 
Production of methyl 4-[3-(2-amino-4-hydroxy-5, 
6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)propyl]benzoate: 
The compound (0.66 g) obtained in the Reference Example 5 was dissolved in 
dried tetrahydrofuran (31.5 ml), to which 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 was added and the resultant precipitate 
was collected by filtration, to give the dihydrobomide of the object 
compound (0.59 g) as a colorless powder. 
IR (KBr): 3290, 3030, 2950, 1720, 1690, 1680 1620, 1480, 1350, 1275, 1100, 
1035, 760 cm.sup.-1. 
.sup.1 H--NMR (DMSO-d.sub.6) .delta.: 1.40-1.83 (4H,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=7.5Hz), 7 86 (2H,d,J=7.5Hz). 
REFERENCE EXAMPLE 7 
Production of diethyl 
N-[4-[3-(2-amino-4-hydroxy-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)propyl] 
benzoyl]-L-g-lutamate: 
The compound (1.47 g) obtained in the Reference Example 6 was suspended in 
tetrahydrofuran (60 ml), to which 0.1 N sodium hydroxide in water (120 ml) 
was added and stirred at room temperature for 21 hours. Then the solution 
was 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), to which diethyl L-glutamate hydrochloride 
(2.88 g), diphenylphosphoryl azide (1.295 ml) and triethylamide (2.52 ml) 
were added, brought back to the room temperature and stirring was 
continued for 63 hours. The resulting precipitate was filtrated off, and 
the filtrate was concentrated to dryness under reduced pressure. The 
residue was subjected to separation-purification with column 
chromatography on silica gel (carrier; 100 g, developing solvent; 
chloroform containing 6.9% ammonia-containing ethanol, 1:20.fwdarw.1:10), 
to give the object compound (1.12 g) as a colorless powder. 
IR (KBr): 3330, 2930, 1740, 1670, 1640, 1570, 1540, 1440, 1375, 1300, 1200, 
1095, 1020 cm.sup.-1. 
.sup.1 H--NMR (CDCl.sub.3 --CD.sub.3 OD) .delta.: 1.20 (3H,t,J=7.5Hz), 1.27 
(3H,t,J=7.5Hz), 1.47-1.83 (4H,m), 2.0-2.36 (2H,m), 2.37-2.50 (2H,m), 2.67 
(2H,t,J=7.5Hz), 3.10-3.37 (2H,m), 3.53-3.80 (1H,m], 3.96-4.33 (4H,q x 
2,J=7.5Hz), 4.60-4.87 (1H,m), 7.25 (2H,d,J=9Hz), 7.75 (2H,d,J=9Hz). 
REFERENCE EXAMPLE 8 
Production of methyl 
4-[3-(2-amino-7-benzyl-3-isopropyloxymethyl-4(3H)-oxopyrrolo[2,3-d]pyrimid 
in-5-yl)-1-oxo-2-propenyl]benzoate: 
2-Amino-7-benzyl-3-isopropyloxymethyl-4(3H)-oxopyrrolo[2,3-d]pyrimidine-5-c 
arbaldehyde (1.7 g) was suspended in a methanol-tetrahydrofuran mixture 
(10:1, 33 ml), to 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 s small amount of methanol and ether, and dried, to give the 
object compound (2.02 g) as yellow needles. 
IR (KBr): 3480, 3350, 1710, 1680, 1620, 1550, 1375, 1280, 1210, 1110, 1060, 
775 cm.sup.-1, 
.sup.1 H--NMR (CDCl.sub.3) .delta.: 1.23 (6H,d,J=6Hz), 3.93 (3H,s), 
3.80-4.07 (1H,m), 5.15 (2H,s), 5.63 (2H,s), 6.92 (1H,s), 7.10-7.40 (5H,m), 
7.73 (1H,d,J=15Hz), 8.13 (4H,s), 8.60 (1H,d,J=15Hz). 
REFERENCE EXAMPLE 9 
Production of methyl 
4-[3-(2-amino-3-isopropyloxymethyl-4(3H)-oxo-5,6-dihydropyrrolo[2,3-d]pyri 
midin-5-yl)propyl]benzoate: 
The compound (2.01 g) obtained in Reference Example 8 was dissolved in a 
methanol-tetrahydrofuran mixture (3:4, 350 ml), to which 1 N hydrochloric 
acid (8ml) and 10% Pd-C (4 g, manufactured by Engelhard Co. Ltd.) were 
added, and subjected to catalytic reduction under an atmosphere of 
hydrogen for 48 hours. The catalyst was filtrated off, the filtrate was 
neutralized, the solvent was evaporated off under reduced pressure, and 
the residue was isolated and purified by column chromatography on silica 
gel (carrier; 100 g) (developing solvent: chloroform containing 2-4% of 
ethanol), to give the object compound (0.68 g) as a colorless powder. 
IR (KBr): 3210, 2980, 1725, 1625, 1580, 1510, 1435, 1275, 1175, 1100, 1060 
cm.sup.-1. 
.sup.1 H--NMR (CDCl.sub.3) .delta.: 1.17 (3H,d,J=6Hz), 1.19(3H,d,J=6Hz), 
1.50-2.13 (4H,m), 2.70 (2H,t,J=7.5Hz), 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 
Production of methyl 
4-[3-(2-amino-4-hydroxy-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)propyl]ben 
zoate: 
The compound (0.66 g) obtained in the Reference Example 9 was dissolved in 
dried tetrahydrofuran (31.5 ml}, to which 0.21 N hydrogen bromide acid in 
dichloromethane (78.3 ml) was added, and stirred at room temperature for 
20 hours. Then 3 volumes of n-hexane was added and the resultant 
precipitate was collected by filtration, to give the dihydrobromide of the 
object compound (0.59 g) as a colorless powder. 
IR (KBr): 3290, 3030, 2950, 1720, 1690, 1680, 1620, 1480, 1350, 1275, 1100, 
1035, 760 cm.sup.-1. 
.sup.1 H--NMR (DMSO-d.sub.6) .delta.: 1.40-1.83 (4H, 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=7.5Hz), 7.86 (2H,d,J=7.5Hz). 
REFERENCE EXAMPLE 11 
Production of methyl 
4-[2-(2-amino-7-benzyl-3-isopropyloxymethyl-4(3H)-oxopyrrolo[2,3-d]pyrimid 
in-5-yl)-ethenyl]benzoate: 
To the suspension of 
2-amino-7-benzyl-3-isopropyloxymethyl-4(3H)-oxopyrrolo[2,3-d]pyrimidine-5- 
carbaldehyde (2.04 g) in dried methanol (84 ml) was added 
p-methoxycarbonylbenzyltriphenylphosphonium bromide (3.24 g) and stirred. 
Then a solution of sodium methylate in methanol (equivalent to 6.6 mM on 
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 object product 
(cis-form, 1.49 g). The mother liquor was purified by column 
chromatography on silica gel (carrier: 100 g) (developing solvent: ethyl 
acetate-hexane, 1:4.fwdarw.1:3), to give the cis-trans mixture of the 
object compound (0.9 g) as yellow powders. cis-form; 
IR (KBr): 3340, 3220, 2980, 1715, 1690, 1625, 1600, 1530, 1430, 1280, 1175, 
1105, 1060, 995 cm.sup.-1. 
.sup.1 H--NMR (CDCL.sub.3) .delta.: 1.20 (6H,d,J=6Hz), 3.87 (3H,s), 
3.80-4.07 (1H,m), 5.14 (2H,s), 5.32 (2H,s), 5.60 (2H,s), 6.77 (1H,s), 
7.10-7.37 (5H,m), 7.43 (2H,s), 7.50 (2H,d,J=9Hz), 7.95 (2H,d,J=9Hz). 
REFERENCE EXAMPLE 12 
Production of methyl 4-[2-(2-amino-3-isopropyloxy 
methyl-4(3H)-oxo-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)ethyl]benzoate: 
The compound (1.6 g) obtained in the Reference Example 11 was subjected to 
the same reaction to that in the Reference Example 9, to give the object 
compound (0.62 g). 
.sup.1 H--NMR (CDCl.sub.3) .delta.: 1.17 (3H,d,J=6Hz), 1.20 (3H,d,J=6Hz), 
1.47-2.0 (1H,m), 2.10-2.43 (1H,m), 2.65 (2H,t,J=9Hz), 2.97-3.60 (3H,m), 
3.73-4.07 (1H,m), 3.90 (3H,s), 4.47 (2H,s), 5.30 (1H,d,J=12Hz), 5.60 
(1H,d,J=12Hz), 7.13-7.50 (7H,m), 7.92 (1H,d,J=9Hz). 
REFERENCE EXAMPLE 13 
Production of methyl 
4-[2-(2-amino-4-hydroxy-7H-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)ethyl]b 
enzoate: 
The compound (1.25 g) obtained in the Reference Example 12 was subjected to 
the same reaction to that in the Reference Example 10, to give the object 
compound (0.51 g). 
IR (KBr): 3400, 3300, 2920, 1740, 1710, 1680, 1640, 1600, 1570, 1435, 1310, 
1280, 1110, 1020 cm.sup.-1. 
.sup.1 H--NMR (DMSO-d.sub.6 /D.sub.2 O) .delta.: 1.53-2.27 (2H,m), 2.70 
(2H,t,J=9Hz), 3.00-3.26 (2H,m) 3.47-3.63 (1H,m), 3.83 (3H,s), 7.35 
(2H,d,J=9Hz), 7.85 (2H,d,J=9Hz) 
REFERENCE EXAMPLE 14 
Production of methyl 5-[4-(tert-butoxycarbonyl)-phenyl]pentanoate: 
Under an atmosphere of argon, potassium (25 g) was dissolved completely in 
dried tert-butyl alcohol (820 ml) by refluxing by heating for 3 hours. The 
solution was cooled to 20.degree. C., to which was added ether (300 ml) 
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 keeping the inner temperature at 10.degree. 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 off under reduced 
pressure. The residue was dissolved in ethyl acetate (100 ml), to which 
was added 5% Pd-C (15 g: manufactured by Engelhard Co. Ltd.), and stirred 
vigorously at room temperature under hydrogen pressure of 4 kg/cm.sup.2 
for 3 hours. The catalyst was filtered off, the solvent was evaporated off 
under reduced pressure, and dried methanol (200 ml), 
4-(N,N-dimethylamino)pyridine (30 mg) and dichloromethane (250 ml) were 
added, to which a solution of 1,3-dicyclohexylcarbodiimide (132 g) in 
dichloromethane (250 ml) was added slowly dropwise at 0.degree. C. After 
stirring at room temperature for 18 hours, the mixture was cooled to 
0.degree. C. Acetic acid (30 ml) was added and the mixture stirred at 
0.degree. 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 was added ethyl 
acetate (100 ml), which was left standing at 0.degree. C. for 2 hours, and 
the resulting precipitate was again filtered off. The filtrate was 
concentrated under reduced pressure, and the residue was purified by 
column chromatography (carrier: silica gel, 500 g, ether-hexane, 
1:15.fwdarw.1:5), to give the object compound (59.7 g). 
Bp. 145.degree.-155.degree. C./0.2-0.3 mmHg. 
IR (Neat): 2980, 2950, 1740, 1712, 1605 cm.sup.-1. 
.sup.1 H--NMR (CDCL.sub.3) .delta.: 1.40-1.75 (4H,m), 1.55 (9H,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 
Production of methyl 5-[4-(tert-butoxycarbnyl)-phenyl]-2-iodopentanoate: 
Under an atmosphere of argon, 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.degree. C. and stirred for 10 minutes. To 
the resultant solution was added a solution of the compound (6.53 g) 
obtained in the Reference Example 14 in tetrahydrofuran (50 ml) at 
-78.degree. C. dropwise over 30 minutes. After stirring for 30 minutes a 
solution of iodine (5.66 g) in tetrahydrofuran (30 ml) was added and 
stirred for further 20 minutes. The temperature was increased to 0.degree. 
C. over 30 minutes, 30 ml of 1 N potassium hydrogen sulfate in water was 
added dropwise, and the pH was adjusted to 4, followed by extraction with 
ether. 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 solvent was evaporated off under reduced pressure, 
and the resultant residue was purified by column chromatography (carrier: 
silica gel, 100 g, ether-hexane, 1:9), to give the object compound (4.736 
g). 
IR (Neat): 2990, 2905, 1744, 1718, 1612 cm.sup.-1. 
.sup.1 H--NMR (CDCl.sub.3) .delta.: 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 (1H,t,J=7Hz), 
7.20(2H,d,J=8Hz), 7.90(2H,d,J=8Hz). 
REFERENCE EXAMPLE 16 
Production of methyl 
5-[4-(tert-butoxycarbonyl)-phenyl]-2-(dicyanomethyl)pentanoate: 
To a suspension of sodium hydride (1.356 g) in dimethylsulfoxide (8 ml) was 
added a solution of malononitrile (3.37 g) in dimethylsulfoxide (8 ml) 
with ice-cooling and stirred for 15 minutes. To the solution was added a 
solution of the compound (4.736 g) obtained in the Reference Example 15 in 
dimethylsulfoxide (12 ml) 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 0.degree. 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 (carrier: silica gel, 200 
g, ethyl acetate-hexane, 1:5), to give the object compound (3.33 g). 
IR (Neat): 2970, 2930, 2252, 1740, 1713, 1608cm.sup.-1. 
.sup.1 H--NMR (CDCl.sub.3) .delta.: 1.60-2.05 (4H,m), 1.48 (9H,s), 2.70 
(2H,brt,J=7Hz), 2.90-3.15 (1H,m), 3.82(3H,s), 4.04(1H,d,J=7Hz), 
7.20(2H,d,J=8Hz), 7.92(2H,d,J=8Hz). 
REFERENCE EXAMPLE 17 
Production 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) was suspended zinc 
(11.81 g), to which ethyl 4-bromocrotonate (17.44 g) was added slowly 
while heating and stirring, and then iodine (about 20 mg) was added. The 
resulting mixture was refluxed by heating on an oil bath 
(60.degree.-70.degree. C.) for 1 hour, then ethyl 4-bromocrotonate (3.00 
g) was added, and the mixture was further refluxed by heating for 15 
minutes. After cooling to room temperature, the reaction mixture was added 
to water (500 ml), adjusted to pH 4.9 by 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 (carrier; silica gel, 300 g, 
developing solvent; ethyl acetate-hexane=1:5), to give the object 
compound. 
IR (Neat): 3480, 2975, 1720, 1700, 1650, 1605 cm.sup.-1. 
.sup.1 H--NMR (CDCl.sub.3) .delta.: 1.20 (3H,t,J=7Hz), 1.53 (12H,s), 2.64 
(2H,d,J=7Hz), 2.67 (1H,brs),3.63 (3H,s), 4.08 (2H,q,J=7Hz), 5.80 
(1H,d,J=15Hz), 6.80 (1H,dt,J=15Hz,7Hz), 7.45 (2H,d,J=8Hz), 7.90 
(2H,d,J=8Hz). 
The entire amount of ethyl hexenate derivative (22.3 g) was dissolved in an 
ethanol-acetic acid mixture (20:1, ml), to which 5% Pd-C (5.0 g) was 
added, then was stirred vigorously for 115 hours. After filtration of Pd-C 
using celite and evaporation of solvent under reduced pressure, the 
residue was subjected to evaporation under reduced pressure, to give the 
object compound (15.66 g) as a colorless oil. 
Bp. 162.degree.-165.degree. C./0.3mmHg 
IR (Neat): 2980, 2940, 1735, 1710, 1607, 848 cm.sup.-1. 
.sup.1 H--NMR (CDCL.sub.3) .delta.: 1.20 (3H,t,J=7Hz), 1.23 (3H,d,J=6Hz), 
1.30-1.80 (4H,m), 1.58 (9H,s), 2.24 (2H,brt,J=6Hz), 2.77 (1H,dq,J=6Hz, 
6Hz), 4.08 (2H,q,J=7Hz), 7.20 (2H,d,J=8Hz), 7.90 (2H,d,J=8Hz). 
REFERENCE EXAMPLE 18 
Production of ethyl 5-[4-(tert-butoxycarbonyl)phenyl]-2-iodohexanate: 
The compound (6.41 g) obtained in the Reference Example 17 was subjected to 
the same reaction as that in the Reference Example 2 to give the object 
compound (3.90 g). 
IR (Neat): 2980, 2940, 1738, 1715, 1610, 850 cm.sup.-1. 
.sup.1 H--NMR (CDCL.sub.3) .delta.: 1.23 (3H, t,J=7Hz), 1.23 (2H,d,J=7Hz), 
1.40-1.95 (4H,m), 1.60 (9H,s), 2.75 (1H,dq,J=6Hz,6Hz), 4.15 (2H,q,J=7Hz), 
4.18 (1H,t,J=7Hz), 7.20 (2H,d,J=8Hz), 7.90 (2H,d,J=8Hz). 
REFERENCE EXAMPLE 19 
Production of ethyl 
5-[4-(tert-butoxycarbonyl)-phenyl]-2-(dicianomethyl)-hexanoate: 
The compound (3.90 g) obtained in the Reference Example 18 was subjected to 
the same reaction as that in the Reference Example 3 to give the object 
compound (3.19 g). 
IR (Neat): 2980, 2930, 2250, 1735, 1710, 1605, 847 cm.sup.-1. 
.sup.1 H--NMR (CDCl.sub.3) .delta.: 1.26 (1.5H,t,J=7Hz), 1.26 (3H,d,J=7Hz), 
1.27 (1.5H,t,J=7Hz), 1.35-1.80 (4H,m), 1.58 (9H,s), 2.50-3.08 (2H,m), 4.00 
(1H,dd,J=8Hz,4Hz), 4.22 (1H,q,J=7Hz), 4.23 (1H,q,J=7Hz), 7.18 
(2H,d,J=8Hz), 7.92 (2H,d,J=8Hz). 
WORKING EXAMPLE 1 
Production of tert-butyl 
4-[3-(2,4-diamino-6-hydroxy-7H-pyrrolo[2,3-d]pyrimidin-5-yl)propyl]benzoat 
e: 
To a solution of potassium tert-butoxide (2.35 g) and guanidine 
hydrochloride (1.07 g) in tert-butylalcohol (10 ml) was added a solution 
of the compound (3.33 g) obtained in the Reference Example 3 in tert-butyl 
alcohol (30 ml) under an atmosphere of argon, and refluxed by heating for 
20 hours. To the reaction mixture were added further potassium 
tert-butoxide (434 mg) and guanidine hydrochloride (370 mg) and the 
mixture was refluxed by heating for 8 further hours. The reaction mixture 
was cooled, added to 1 N potassium hydrogen sulfate in water (about 10 ml) 
and adjusted to pH 9. After extraction with a tetrahydrofuran-chloroform 
mixture, the solvent was evaporated off under reduced pressure and the 
resultant residue was purified by column chromatography (carrier; silica 
gel, 100 g, developing solvent; dichloromethane: ethanol 
=15:1.fwdarw.dichloromethane after mixing with concentrated aqueous 
ammonia in a separatory funnel: ethanol =15:1), to give the object 
compound (1.90 g). 
IR (KBr): 3430, 3360, 1710, 1627, 1583, 1432 cm.sup.-1. 
.sup.1 H--NMR (CDCl.sub.3 -Me.sub.2 SO-d.sub.6) .delta.: 1.15-1.73 (2H,m), 
1.55 (9H,s), 1.73-2.10 (2H,m), 2.61 (2H,t,J=7Hz), 3.35 (1H,t,J=6Hz), 5.40 
(2H,brs), 5.51 (2H,brs), 6.30 (1H,brs), 7.12 (2H,d,J=8Hz), 7.29 
(2H,d,J=8Hz). 
WORKING EXAMPLE 2 
Production of tert-butyl 
4-[3-(2,4-diamino-7H-pyrrolo[2,3-d]pyrimidin-5-yl)propyl]benzoate: 
To a solution of the compound (575 mg) obtained in the Working Example 1 in 
tetrahydrofuran (6 ml) was added a solution of borane-tetrahydrofuran 
complex (7.5 mmol) in tetrahydrofuran (7.5 ml) at 0.degree. 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 off under reduced pressure and 
the residue was purified by column chromatography (carrier; silica gel, 30 
g, developing solvent; dichloromethane: ethanol =100:6 
100:7.fwdarw.100:8.fwdarw.10:1.fwdarw.8:1), to give the object compound 
(263 mg). 
IR (KBr): 3335, 3180, 2975, 2935, 1710, 1607, 1287, 1163, 1110 cm.sup.-1. 
.sup.1 H--NMR (Me.sub.2 SO-d.sub.6) .delta.: 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 
(1H,s), 7.33 (2H,d,J=8Hz), 7.82 (2H,d,J=8Hz), 10.51 (1H,s). 
WORKING EXAMPLE 3 
Production of diethyl 
N-[4-[3-(2,4-diamino-7H-pyrrolo[2,3-d]pyrimidin-5-yl)propyl]benzoyl]-L-glu 
tamate: 
To the compound (381 mg) obtained in the Working Example 2 was added 
trifluoroacetic acid (3 ml) and the mixture was stirred at room 
temperature for 3 hours. Trifluoroacetic acid was evaporated off under 
reduced pressure, and to the residue obtained by drying at 70.degree. C. 
under reduced pressure and a solution of diethyl L-glutamate hydrochloride 
(748 mg) in dimethylformamide (4 ml) was added a solution of 
diphenylphosphoryl azide (858 mg) in dimethylformamide (4 ml) at 0.degree. 
C., and then a solution of triethylamine (631 mg) in dimethylformamide (4 
ml) dropwise at the same temperature. After stirring at 0.degree. C. for 
30 minutes and then at room temperature for 63 hours, the precipitate was 
filtered off. The solvent was evaporated off under reduced pressure and 
the resultant residue was purified by column chromatography (carrier; 
silica gel, 20 g, developing solvent; dichloromethane after mixing with 
concentrated aqueous ammonia in a separatory funnel.fwdarw.dichloromethane 
after mixing with concentrated aqueous ammonia: ethanol 
=40:1.fwdarw.30:1), to give the object compound (260 mg). 
IR (KBr): 3330, 3160, 1735, 1632, 1575, 1540, 1500, 1200 cm.sup.-1. 
.sup.1 H--NMR (Me.sub.2 SO-d.sub.6) .delta.: 1.17(3H,t,J=7Hz), 1.20 
(3H,t,J=7Hz), 1.80-2.20 (4H,m), 2.44 (2H,t,J=7Hz), 2.68 (2H,t,J=7Hz), 2.72 
(2H,t,J=7Hz), 4.05 (2H,q,J=7Hz), 4.11 (2H,q,J=7Hz), 4.35-4.50 (1H,m), 5.34 
(2H,s), 5.91 (2H,s), 6.42 (1H,s), 7.31 (2H,d,J=8Hz), 7.80 (2H,d,J=8Hz), 
8.66 (1H,d,J=8Hz), 10.51 (1H,s). 
WORKING EXAMPLE 4 
Production of N-[4-[3-(2,4-diamino-7H-pyrrolo 
[2,3-d]pyrimidin-5-yl)propyl]benzoyl]-L-glutamic acid: 
The compound (250 mg) obtained in the Working Example 3 was dissolved in 
tetrahydrofuran-water mixture (1:1, 7 ml), to which 1 N sodium hydroxide 
in water (2.52 ml) was added and the mixture was stirred at room 
temperature for 1.5 hours. The solution was concentrated to 3 ml under 
reduced pressure, and the resultant insoluble matter was filtrated off 
through a millipore filter. To the filtrate, cooled to 0.degree. C., was 
added acetic acid (0.5 ml) and the resultant crystals were collected by 
filtration and washed throughly with ice water. The crystals were dried at 
70.degree. C. under reduced pressure, to give the object compound (201 mg) 
as white crystals. 
IR (KBr): 3340, 3200, 2940, 1660-1630, 1540, 1500, 1397 cm.sup.-1. 
.sup.1 H--NMR (Me.sub.2 SO-d.sub.6) .delta.: 1.75-2.20 (4H,m), 2.35 
(2H,t,J=7Hz), 2.68 (2H,t,J=7Hz), 2.71 (2H,t,J=7Hz), 4.30-4.47 (1H,m), 5.53 
(2H,brs), 6.15 (2H,s), 6.46 (1H,s), 7.31 (2H,d,J=8Hz), 7.81 (2H,d,J=8Hz), 
8.48 (1H,d,J=8Hz), 10.51 (1H,s). 
WORKING EXAMPLE 5 
Production of diethyl 
N-[4-[3-(2,4-diamino-6-hydroxy-7H-pyrrolo[2,3-d]pyrimidin-5-yl) 
propyl]benzoyl]-L-glutamate: 
The compound (200 mg) obtained in the Working Example was subjected to the 
same reaction as that in the Working Example 3, to give the object 
compound (164 mg). 
IR (KBr): 3355, 3230, 2994, 2990, 1740, 1638, 1595, 1590 cm.sup.-1. 
.sup.1 H--NMR (CDCl.sub.3 /Me.sub.2 SO-d.sub.6) .delta.: 1.20 (3H,t,J=7Hz), 
1.25 (3H,t,J=7Hz), 1.25-2.70 (11H,m), 3.25-3.45 (1H,m), 4.05 (2H,q,J=7Hz), 
4.15 (2H,q,J=7Hz), 4.38-4.68 (1H,m), 5.63 (2H,brs), 5.66 (2H,brs), 7.16 
(2H,d,J=8Hz), 7.76 (2H,d,J=8Hz), 8.39 (1H,d,J=8Hz), 10.50 (1H,s). 
WORKING EXAMPLE 6 
Production of 
N-[4-[3-(2,4-diamino-6-hydroxy-7H-pyrrolo[2,3-d]pyrimidin-5-yl)propyl]benz 
oyl]-L-glutamic acid: 
The compound (112 mg) obtained in the Working Example was subjected to the 
same reaction as that in the Working Example 4, to give the object 
compound (60 mg). 
IR (KBr): 3350, 3210, 2950, 1730, 1660, 1630 cm.sup.-1. 
.sup.1 H--NMR (Me.sub.2 SO-d.sub.6) .delta.: 1.20-1.56 (2H,m], 
1.65-2.20(4H,m), 2.35 (2H,t,J=7Hz), 2.50-2.65 (2H,m), 3.25-3.35 (1H,m), 
4.32-4.46 (1H,m), 5.90 (2H,brs), 6.00 (2H,brs). 7.22 (2H,d,J=8Hz), 7.78 
(2H,d,J=8Hz), 8.52 (1H,d,J=8Hz), 10.45 (1H,s). 
WORKING EXAMPLE 7 
Production of diethyl 
N-[4-[3-(2-amino-4-hydroxy-7H-pyrrolo[2,3-d]pyrimidin-5-yl)propyl]benzoyl] 
-L-glutamate: 
The compound (150 mg) obtained in the Reference Example 7 was dissolved in 
ethanol (22.5 ml), to which were added 10% Pd-C (450 mg: manufactured by 
Engelhand Co. Ltd.) and 2 drops of acetic acid and stirred vigorously at 
room temperature for 62.5 hours. The catalyst was filtered off, the 
filtrate was concentrated to dryness and the residue was purified by 
column chromatography (carrier; silica gel, 10 g, developing solvent; 
chloroform containing 5% ethanol), to give the object compound (33 mg). 
IR (KBr): 3340, 2940, 1740, 1680, 1670, 1630, 1540, 1440, 1380, 1340, 1210, 
1100, 1020, 860 cm.sup.-1. 
.sup.1 H--NMR (CDCl.sub.3 /CD.sub.3 OD) .delta.: 1.20 (3H,t,J=6Hz), 1.28 
(3H,t,J=6Hz), 1.87-2.36 (4H,m), 2.40-2.57 (2H,m), 2.60-2.87 (4H,m), 
3.96-4.37 (4H,q x 2,J=6Hz), 4.56-4.90 (1H,m), 6.37 (1H,s), 7.23 
(2H,d,J=7.5Hz), 7.71 (2H,d,J=7.5Hz). 
WORKING EXAMPLE 8 
Production of 
N-[4-[3-(2-amino-4-hydroxy-7H-pyrrolo[2,3-d]pyrimidin-5-yl)propyl]benzoyl] 
-L-glutamic acid: 
The compound (27 mg) obtained in the Working Example 7 was dissolved in a 
tetrahydrofuran-water mixture (1:1, 2.16 ml), to which was added 1 N 
sodium hydroxide in water (0.189 ml) and stirred at room temperature for 
2.5 hours. A large portion of tetrahydrofuran was evaporated off, acetic 
acid (0.189 ml) was added by ice-cooling and stirred. The resulting 
precipitate was collected by filtration and dried, to give the object 
compound (19 mg). 
IR (KBr): 3400, 3300, 2950, 1700, 1650, 1540, 1510, 1400, 1340, 1240, 1080, 
1020 cm.sup.-1 
.sup.1 H--NMR (Me.sub.2 SO-d.sub.6) .delta.: 1.80-2.17 (4H,m), 2.23-2.40 
(2H,m), 2.53-2.83 (4H,m), 4.27-4.56 (1H,m), 6.33 (1H,s), 7.27 
(2H,d,J=7.5Hz), 7.78 (2H,d,J=7.5Hz). 
WORKING EXAMPLE 9 
Production of tert-butyl 
4-[3-(2,4-diamino-6-oxo-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)propyl]ben 
zoate: 
To a solution of potassium tert-butoxide (2.35 g) and guanidine 
hyrochloride (1.07 g) in tert-butyl alcohol (10 ml) was added a solution 
of the compound (3.33 g) obtained in the Reference Example 16 in 
tert-butyl alcohol (30 ml) under an atmosphere of argon, and refluxed by 
heating for 20 hours. To the reaction mixture were added further potassium 
tert-butoxide (434 mg) and guanidine hydrochloride (370 mg), and refluxed 
by heating for further 8 hours. The reaction mixture was cooled, added to 
1 N potassium hydrogen sulfate in water (about 10 ml), to be adjusted to 
pH 9. After extraction with a tetrahydrofuran-chloroform mixture, the 
solvent was evaporated off under reduced pressure, and the resultant 
residue was purified by column chromatography (carrier; 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 object compound (1.90 g). 
IR (KBr): 3430, 3360, 1710, 1627, 1583, 1432cm.sup.-1. 
.sup.1 H--NMR (CDCl.sub.3 /Me.sub.2 SO-d.sub.6) .delta.: 1.15-1.73 (2H,m), 
1.55 (9H,s), 1.73-2.10 (2H,m), 2.61 (2H,t,J=7Hz), 3.35 (1H,t,J=6Hz), 5.40 
(2H,brs), 5.51 (2H,brs), 6.30 (1H,brs), 7.12 (2H,d,J=8Hz), 
7.29(2H,d,J=8Hz). 
WORKING EXAMPLE 10 
Production 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) obtained in the Working Example 9 in 
tetrahydrofuran (10 ml) was added a solution of borane-tetrahydrofuran 
complex (16.8 mmol) in tetrahydrofuran (10 ml) and the mixture was 
refluxed by heating for 4 hours. After cooling, the reaction mixture was 
added to ice water and stirred vigorously at pH 2 (adjusted by addition of 
1N hydrochloric acid) for 3 minutes and then 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 off under reduced pressure, and the residue was purified by 
column chromatography (carrier; silica gel, 20 g, dichloromethane-ethanol, 
30:1.fwdarw.15:1 dichloromethane after mixing with concentrated aqueous 
ammonia in a separatory funnel-ethanol, 20:1), to give the object compound 
(114 mg). IR (KBr): 3375, 3325, 3190, 2970, 2930, 1712, 1603 cm.sup.-1. 
.sup.1 H--NMR (CDCl.sub.3 /Me.sub.2 SO-d.sub.6) .delta.: 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 (1H,m), 4.85 
(2H,brs), 4.90 (2H,brs), 5.30 (1H,brs), 7.19 (2H,d,J=8Hz), 7.80 
(2H,d,J=8Hz). 
WORKING EXAMPLE 11 
Production of diethyl 
N-[4-[3-(2-amino-4-hydroxy5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)propyl]b 
enzoyl]-L-glutamate: 
The compound (1.47 g) obtained in the Reference Example 10 was suspended in 
tetrahydrofuran (60 ml), to which 0.1 N sodium hydroxide in water (120 ml) 
was added, and the mixture was stirred at room temperature for 21 hours. 
Then the solution was 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), to which diethyl L-glutamate 
hydrochloride (2.88 g), diphenylphosphor-yl azide (1.295 ml) and 
triethylamine (2.52 ml) were added. The mixture was brought back to the 
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 with column chromatography on silica gel (carrier; 
100 g) (ethanol containing 6.9% ammonia-chloroform containing 6.9% 
ammonia, 1:20.fwdarw.1:10), to give the object compound (1.12 g) as a 
colorless powder. 
IR (KBr): 3330, 2930, 1740, 1670, 1640, 1570, 1540, 1440, 1375, 1300, 1200, 
1095, 1020 cm.sup.-1. 
.sup.1 H--NMR (CDCl.sub.3 /CD.sub.3 OD).delta.: 1.20 (3H,t,J=7.5Hz), 1.27 
(3H,t,J=7.5Hz), 1.47-1.83 (4H,m), 2.00-2.36 (2H,m), 2.37-2.50 (2H,m), 2.67 
(2H,t,J=7.5Hz), 3.10-3.37 (2H,m), 3.53-3.80 (1H,m), 3.96-4.33 
(4H,qx2,J=7.5Hz), 4.60-4.87 (1H,m), 7.25 (2H,d,J=9Hz), 7.75 (2H,d,J=9Hz). 
WORKING EXAMPLE 12 
Production of diethyl 
N-[4-[2-(2-amino-4-hydroxy5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)ethyl]be 
nzoyl]-L-glutamate: 
The compound (0.315 g) obtained in the Reference Example 13 was subjected 
to the same reaction as that in the Working Example 11, to give the object 
compound (0.247 g). 
IR (KBr): 3310, 2990, 1740, 1730, 1690, 1640, 1570, 1530, 1440, 1375, 1330, 
1300, 1240, 1200, 1090, 1010, 850cm.sup.-1. 
.sup.1 H--NMR (CDCl.sub.3 /CD.sub.3 OD).delta.: 1.22 (3H,t,J=7.5Hz), 1.30 
(3H,t,J=7.5Hz), 1.53-2.87 (8H,m), 3.13-3.90 (3H,m), 4.00-4.43 
(4H,qx2,J=7.5Hz), 4.57-4.90 (1H,m), 7.25 (2H,d,J=9Hz), 7.72 (2H,d,J=9Hz). 
WORKING EXAMPLE 13 
Production of diethyl 
N-[4-[3-(2,4-diamino-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)propyl]benzoy 
l]-L-glutamate: 
The compound (94 mg) obtained in the Working Example 10 was dissolved in 1 
ml of trifluoroacetic acid and stirred at room temperature for 3 hours. 
The solvent was evaporated off under reduced pressure, and to the mixture 
of the residue obtained by drying at 70.degree. C. under reduced pressure 
and diethyl L-glutamate (304 mg) in dimethylformamide (2 ml), was added a 
solution of diphenylphosphoryl azide (350 mg) in dimethylformamide (1.5 
ml) at 0.degree. C., and then a solution of triethylamine (180 mg) in 
dimethylformamide (1.5 ml) dropwise at the same temperature. After 
stirring at 0.degree. C. for 30 minutes and then at room temperature for 
78 hours, the solvent was evaporated off under reduced pressure. The 
resultant residue was purified by column chromatography (carrier; silica 
gel, 20 g, dichloromethane after mixing with concentrated aqueous ammonia 
in a separatory funnel-dichloromethane after mixing with concentrated 
aqueous ammonia -ethanol 40:1.fwdarw.30:1), to give the object compound 
(88 mg). 
IR (KBr): 3350, 2990, 2945, 1740, 1610, 1540, 1508, 1438cm.sup.-1. 
.sup.1 H--NMR (CDCl.sub.3) .delta.: 1.23 (H,q,J=7Hz), 1.43-1.80 (3H,m), 
1.85-2.77 (7H,m), 2.95-3.30 (2H,m), 3.58 (1H,t,J=11Hz), 4.07 (2H,q,J=7Hz), 
4.20 (2H,q,J=7Hz), 4.25 (1H,brs), 4.63-4.83 (1H,m), 4.68 (1H,brs), 
7.00-7.23 (1H,m), 7.13 (2H,d,J=8Hz), 7.67 (2H,d,J=8Hz). 
WORKING EXAMPLE 14 
Production of 
N-[4-[3-(2-amino-4-hydroxy-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)propyl] 
benzoyl]-L-glutamic acid: 
The compound (1.05 g) obtained in the Working Example 11 was dissolved in 
tetrahydrofuran-water mixture (2:1, 63 ml), to which 1 N sodium hydroxide 
in water (7.35 ml) was added, and stirred at room temperature for 2.5 
hours. Tetrahydrofuran was evaporated off, a small amount of insoluble 
matter was filtrated off, acetic acid (7.35 ml) was added to the filtrate, 
and the resulting precipitate was collected by filtration. The precipitate 
was washed with water and dried, to give the object compound (0.85 g) as a 
colorless powder. 
IR (KBr): 3340, 2930, 1690, 1630, 1540, 1440, 1300, 1080, 850 cm.sup.-1. 
.sup.1 H--NMR (DMSO-d.sub.6) .delta.: 1.20-1.80 (4H,m), 1.87-2.17 (2H,m), 
2.23-2.40 (2H,m), 2.50-2.77 (2H,m), 2.83-3.20 (2H,m), 3.30-3.63 (1H,m), 
4.23-4.53 (1H,m), 7.26 (2H,d,J=9Hz), 7.77 (2H,d,J=9Hz). 
WORKING EXAMPLE 15 
Production of 
N-[4-[2-(2-amino-4-hydroxy-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)ethyl]b 
enzoyl]-L-glutamic acid: 
The compound (0.195 g) obtained in the Working Example 12 was subjected to 
the same reaction as that in the Working Example 14, to give the object 
compound (0.153 g). 
IR (KBr): 3250, 2900, 1650, 1580, 1440, 1300, 1090cm.sup.-1. 
.sup.1 H--NMR (DMSO-d.sub.6) .delta.: 1.43-1.76 (1H,m), 1.98 
(2H,t,J=7.5Hz), 1.80-2.10 (1H,m), 2.13-2.40 (2H,m), 2.67 (2H,t,J=9Hz), 
2.90-3.23 (2H,m), 3.33-3.60 (1H,m), 4.10-4.43 (1H,m), 7.28 (2H,d,J=9Hz), 
7.75 (2H,d,J=9Hz). 
WORKING EXAMPLE 16 
Production of 
N-[4-[3-(2,4-diamino-5,6-dihydropyrrolo[2,3-d]pyrimidin-5-yl)propyl]benzoy 
l]-L-glutamic acid: 
The compound (41 mg) obtained in the Working Example was subjected to the 
same reaction as that in the 
WORKING EXAMPLE 14, to give the object compound (32 mg). 
IR (KBr): 3700-2350, 3215, 1690-1620, 1540cm.sup.-1. 
.sup.1 H--NMR (Me.sub.2 SO-d.sub.6) .delta.: 1.02-1.85 (4H,m), 1.85-2.83 
(6H,m), 2.90-3.30 (2H,m), 3.55 (1H,t,J=11Hz), 4.15-4.45 (1H,m), 6.38 
(2H,brs), 6.77 (2H,brs), 6.90 (1H,brs), 7.22 (2H,d,J=8Hz), 7.74 
(2H,d,J=8Hz), 8.22 (1H,d,J=7Hz). 
WORKING EXAMPLE 17 
Production of tert-butyl 
4-[3-(2,4-diamino-6-hydroxy-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-1-methylpropy 
l]benzoate: 
The compound (3.18 g) obtained in the Reference Example 19 was subjected to 
the same reaction as that in the Working Example 1, to give the object 
compound (2.61 
IR (KBr): 3360, 3235, 2975, 2700, 1715, 1625, 1584, 1438, 1290, 1163, 1118, 
848 cm.sup.-1. 
.sup.1 H--NMR (Me.sub.2 SO-d.sub.6) .delta.: 1.14 (3H,d,J=7Hz), 1.20-1.50 
(2H,m), 1.54 (9H,s), 1.55-1.80 (1H,m), 1.80-2.05 (1H,m), 2.60-2.78 (1H,m), 
3.20-3.30 (1H,m), 5.86 (2H,brs), 5.96 (2H,brs), 7.25 (2H,d,J=8Hz), 7.81 
(2H,d,J=8Hz), 10.42 (1H,s). 
WORKING EXAMPLE 18 
Production of tert-butyl 
4-[3-(2,4-diamino-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-5-yl)-1-methylpro 
pyl]-benzoate (A) and 
tert-butyl-4-[3-(2,4-diamino-7H-pyrrolo-[2,3-d]pyrimidin-5-yl)-1-methyl-pr 
opyl]benzoate (B): 
To a suspension of the compound (2.00 g) obtained in the Working Example 17 
in tetrahydrofuran (25 ml) was 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 solvent under reduced pressure, the 
resulting residue was purified by column chromatography (carrier: silica 
gel, 100 g, developing solvent; dichloromethane-ethanol=20:1.fwdarw.25:2, 
then dichloromethane-ethanol containing ammonia (6%)), to give the object 
compound (A) (579 mg) and (B) (1.214 g). 
(A) IR (KBr): 3350, 3200, 2980, 2940m, 1714, 1650, 1608, 1290, 1163, 1115, 
848 cm.sup.-1. 
.sup.1 H--NMR (CDCl.sub.3) .delta.: 1.31 (3H,d,J=7Hz), 1.60 (9H,s), 1.94 
(2H,dt,J=8Hz,8Hz), 2.40-2.60 (2H,m), 2.85 (1H,tq,J=7Hz,7Hz), 4,50-5.50 
(4H,br), 6.46 (1H,s), 7.27 (2H,d,J=8Hz), 7.96 (2H,d,J=8Hz), 9.20 (1H,brs). 
(B) IR (KBr): 3340, 3195, 2980, 2936, 1715, 1607, 1430, 1295, 1163, 1115, 
847 cm.sup.-1. 
.sup.1 H--NMR (Me.sub.2 SO-d.sub.6) .delta.: 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 (9H,s), 
2.64-2.70 (1H,m), 2,90-3.08 (2H,m), 3.30-3.50 (1H,m), 5.43 (4H,s), 5.95 
(0.25H,s), 6.00 (0.75H,s), 7.32 (2H,d,J=8Hz), 7.82 (2H,d,J=8Hz). 
WORKING EXAMPLE 19 
Production of diethy 
N-[4-[3-(2,4-diamino-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-5-yl)-1-methyl 
propyl]-benzoyl]-L-glutamate 
The compound (A) (581 mg) obtained in the Working Example 18 was subjected 
to the same reaction as that in the Working Example 3 to give the object 
compound (640 mg). 
IR (KBr): 3375, 3200, 2975, 2930, 1738, 1608, 1430, 1200, 1008, 853 
cm.sup.-1 
.sup.1 H--NMR (Me.sub.2 SO-d.sub.6) .delta.: 1.17 (3H,t,J=7Hz), 1.18 
(3H,d,J=7Hz), 1.19 (3H,t,J=7Hz), 1.26-1.44 (2H,m), 1.44-1.63 (2H,m), 
1.90-2.20 (2H,m), 2.44 (2H,t,J=7Hz), 2.63-2.80 (1H,m), 2.90-3.08 (2H,m), 
3.30-3.50 (1H,m), 4.05 (2H,q,J=7Hz), 4.11 (2H,q,J=7Hz), 4,37-4.50 (1H,m), 
5.36 (2H, s), 5.37 (2H,s), 5.87 (0.25H,s), 5.91 (0.75H,s), 7.30 
(2H,d,J=8Hz,), 7.80 (2H,d,J=8Hz), 8.66 (1H,d,J=8Hz). 
WORKING EXAMPLE 20 
Production of 
N-[4-[3-(2,4-diamino-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-5-yl)-1-methyl 
propyl]benzoyl]-L-glutamic acid: 
The compound (600 mg) obtained in the Working Example 19 was subjected the 
same reaction as that in the Working Example 4, to give the object 
compound (508 mg). 
IR (KBr): 3350, 3200, 1690, 1680-1610, 1635, 1530, 1400, 1300, 853 
cm.sup.-1. 
.sup.1 H--NMR (Me.sub.2 SO-d.sub.6) .delta.: 1.20 (3H,d,J=7Hz), 1.25-1.65 
(4H,m), 1.87-2.20 (2H,m), 2.30 (2H,t,J=7Hz), 2.60-2.80 (1H,m), 3.00-3.20 
(2H,m), 3.42-3.60 (1H,m), 4.22-4.40 (1H,m), 6.20-4.08 (5H,m), 7.28 
(2H,d,J=8Hz), 7.78 (2H,d,J=8Hz), 8.28-8.36 (1H,m). 
WORKING EXAMPLE 21 
Production of diethyl 
N-[4-[3-(2,4-diamino-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-1-methylpropyl]benzo 
yl]-L-glutamic acid: 
The compound (B) (540 mg) obtained in the Working Example 18 was subjected 
to the same reaction as that in the Working Example 3, to give the object 
compound (556 mg). 
IR (KBr): 3340, 3180, 2935, 1735, 1640, 1610, 1580, 1200, 1095, 1018, 850 
cm.sup.-1. 
.sup.1 H--NMR (CDCl.sub.3) .delta.: 1.23 (3H,t,J=7Hz), 1.30 (3H,d,J=7Hz), 
1.31 (3H,t,J=7Hz), 1.80-2.05 (4H,m), 2.15-2.57 (4H,m), 2.83 
(1H,tq,J=7Hz,7Hz), 4.12 (2H,q,J=7Hz), 4.25 (2H,q,J=7Hz), 4.68 (2H,brs), 
4.75-4.87 (1H,m), 4.92 (2H,brs), 6.43 (1H,s), 7.26 (2H,d,J=8Hz), 7.37 
(1H,dd,J=3Hz), 7.77 (2H,d,J=8Hz), 8.81 (1H,brs). 
WORKING EXAMPLE 22 
Production of 
N-[4-[3-(2,4-diamino-7H-pyrollo[2,3-d]-pyrimidin-5-yl)-1-methylpropyl]benz 
oyl]-L-glutamic acid: 
The compound (533 mg) obtained in the Working Example 21 was subjected to 
the same reaction as that in the Working Example 4, to give the object 
compound (436 mg). 
IR (KBr): 3350, 3205, 1650, 1640, 1540, 1400, 840 cm.sup.-1. 
.sup.1 H--NMR (Me.sub.2 SO-d.sub.6) .delta.: 1.25 (3H,d,J=7Hz), 1.73-2.20 
(4H,m), 2.35 (3H,t,J=8Hz), 2.40-2.68 (2H,m), 2.85 (1H,tq,J=7Hz,7Hz), 
4.32-4.45 (1H,m), 5.54 (2H,brs), 6.06 (2H,brs), 6.38 (1H,s), 7.33 
(2H,d,J=8Hz), 7.83 (2H,d,J=8Hz), 8.49 (1H,d,J=8Hz), 10.45 (1H,s). 
WORKING EXAMPLE 23 
The compound (50 mg per tablet) obtained in the Working Example 14, lactose 
(250 mg per tablet), corn starch (51 mg per tablet) and 
hydroxypropylcellulose L (9 mg per tablet) were mixed according to the 
conventional method and granulated. The granules, corn starch (8 mg per 
tablet) and magnesium stearate (2 mg per tablet) were mixed and tabletted 
according to the conventional method, to give the tablets (370 mg per 
tablet). 
WORKING EXAMPLE 24 
Ten grams of the sodium salt of the compound obtained in the Working 
Example 14 was dissolved in 1l of physiologial saline. The solution was 
filtered through a microfilter and dispensed in 2.2 ml aliquots in 
ampoules, sterilized at 110.degree. C. for 30 minutes and the ampoules may 
be used for subcutaneous, intravenous or intramuscular injections. 
WORKING EXAMPLE 25 
Five grams of the hydrochloride of the compound obtained in the Working 
Example 14 and 10 g of mannitol were dissolved in 1 l of distilled water, 
and the solution was dispensed in 2 ml aliquots into ampules after 
filtration through a bacterial filter. The ampoules were dried in a 
freeze-drier and sealed, and thus the ampoules of which content is 
dissolved before use were obtained. Before use for injection, the ampoules 
are opened and the content is dissolved in, for example, 2 ml of 
physiological saline.