Adenosine deaminase inhibitor

The present invention relates to adenosine deaminase inhibitors containing at least one O-alkylated moiety and the pharmaceutically acceptable salts thereof. The pharmaceutical compositions of the present invention include adenosine deaminase inhibitors containing at least one of the compounds represented by Formula (I): ##STR1## wherein each of R.sub.1, R.sub.2, and R.sub.3 are the same or different and is hydrogen or alkyl; PA1 R is hydrogen, alkyl, alkenyl, alkynyl, hydroxyalkynyl, alkoxy, phenyl, hydroxy, amino, alkylamino, phenylamino or halogen; PA1 X is hydrogen, alkyl, alkynyl, allyl, methallyl, cycloalkyl, alkyl having one or more hydroxy groups, phenyl, substituted phenyl, alkyl having one or more phenyl groups, alkyl having one or more substituted phenyl groups, bicycloalkyl, naphthylalkyl, acenaphthylenylalkyl or a compound represented by Formula (II) or Formula (III) ##STR2## wherein Z is hydrogen, hydroxy or lower alkoxy; Q is hydrogen or hydroxy; PA1 A is --CH.sub.2 --, --O--, --S-- or a mere linkage; PA1 Y is (CH.sub.2).sub.n -- or a mere linkage; PA1 n is an integer from 1 to 3; and PA1 any of R.sub.1, R.sub.2, and R.sub.3 is R.sub.3 lower alkyl.

FIELD OF THE INVENTION 
The present invention relates to adenosine deaminase inhibitors containing 
at least one O-alkylated moiety and the pharmaceutically acceptable salts 
thereof. 
BACKGROUND OF THE INVENTION 
Adenosine deaminase is an enzyme producing inosine by deamination of 
adenosine in vivo and is prevalent in animals and microorganisms. When 
adenosine deaminase is inhibited, the adenosine concentration in tissues 
is increased while the inosine concentration is decreased whereupon 
endogenous inactivation of adenosine is inhibited. When the tissue is in 
an ischemic state, neutrophils produce activated oxygen and adenosine 
inhibits this oxygen production. In addition, adenosine directly 
eliminates the produced activated oxygen. Further, as a result of a 
decrease in the inosine concentration, the supply of hypoxanthine is 
decreased. Hypoxanthine is a substrate in the xanthine-xanthineoxidase 
system. The xanthine-xanthineoxidase system is one of the systems 
producing the activated oxygen. It has been known that adenosine deaminase 
inhibitors, which inhibit the production of such activated oxygen sources 
and also eliminate them, exhibit pharmacological actions such as 
improvement of coronary and cerebral blood vessel circulation, prevention 
and therapy of renal diseases, and antiinflammatory activity. 
It has been found that the O-alkylated adenosine derivatives of the instant 
invention exhibit excellent adenosine deaminase inhibiting action. 
SUMMARY OF THE INVENTION 
The present invention pertains to adenosine deaminase inhibitors containing 
at least one O-alkylated moiety and the pharmaceutically acceptable salts 
thereof. 
The pharmaceutical compositions of the present invention include adenosine 
deaminase inihibitors containing at least one of the compounds represented 
by Formula (I): 
##STR3## 
wherein each of R.sub.1, R.sub.2, and R.sub.3 are the same or different 
and is hydrogen or alkyl; 
R is hydrogen, alkyl, alkenyl, alkynyl, hydroxyalkynyl, alkoxy, phenyl, 
hydroxy, amino, alkylamino, phenylamino or halogen; 
X is hydrogen, alkyl, alkynyl, allyl, methallyl, cycloalkyl, alkyl having 
one or more hydroxy groups, phenyl, substituted phenyl, alkyl having one 
or more phenyl groups, alkyl having one or more substituted phenyl groups, 
bicycloalkyl, naphthylalkyl, acenaphthylenylalkyl or a compound 
represented by Formula (II) or Formula (III) 
##STR4## 
wherein Z is hydrogen, hydroxy or lower alkoxy; Q is hydrogen or hydroxy; 
A is --CH.sub.2 --, --O--, --S-- or a single bond forming a five-membered 
ring; 
Y is (CH.sub.2).sub.n -- or a single bond; 
n is an integer from 1 to 3; and at least one of R.sub.1, R.sub.2, and 
R.sub.3 is alkyl, such as a lower alkyl. 
The compounds represented by Formula (I) are present in the adenosine 
deaminase inhibitors in a pharmaceutically effective amount. 
The compounds of the present invention having adenosine deaminase 
inhibiting action are useful pharmaceutical compositions for the 
prevention and therapy of various kinds of diseases. Such diseases include 
ischemic heart diseases, diseases caused by cerebrovascular disorder, 
renal diseases and allergic diseases. Moreover, the compounds of the 
present invention are very useful pharmaceutical compositions for the 
prevention and therapy of post-operative complicated diseases because they 
inactivate activated oxygen which is generated in ischemic areas during 
the recirculation of blood after operations. 
The compounds of the instant invention may also be administered before or 
together with aniticancer drugs and/or antiviral drugs. 
DETAILED DESCRIPTION OF THE INVENTION 
The present invention relates to adenosine deaminase inhibitors containing 
a pharmaceutically effective amount of at least one of the compounds 
represented by the following general formula (I) or pharmaceutically 
acceptable salts thereof. 
##STR5## 
In Formula (I) each of R.sub.1, R.sub.2 and R.sub.3 may be the same or 
different and is hydrogen or alkyl; R is hydrogen, alkyl, alkenyl, 
alkynyl, hydroxyalkynyl, alkoxy, phenyl, hydroxy, amino, alkylamino, 
phenylamino or halogen; X is hydrogen, alkyl, alkynyl, allyl, methallyl, 
cycloalkyl, alkyl having one or more hydroxy groups, phenyl, substituted 
phenyl, alkyl having one or more phenyl groups, alkyl having one or more 
substituted phenyl groups, bicycloalkyl, naphthylalkyl, 
acenaphthylenylalkyl or a group represented by the following general 
formulae (II) or (III): 
##STR6## 
Z is hydrogen, hydroxy or lower alkoxy; Q is hydrogen or hydroxy; A is 
--CH.sub.2 --, --O--, --S-- or a single bond forming a five-membered ring; 
Y is --(CH.sub.2)n-- or a single bond; n is an integer from 1 to 3 and at 
least one of R.sub.1, R.sub.2 and R.sub.3 is an alkyl group. 
A substituted phenyl or a substituted phenyl group is defined, for the 
purposes of this invention, as including a phenyl which has been 
substituted with one or more halogen, lower alkyl, lower alkoxy and/or 
trifluoromethyl substituents. In Formula (II), a broken line symbolizes 
the presence of either a double bond or a single bond. 
In the above general formula (I), each of R.sub.1, R.sub.2 and R.sub.3 may 
be the same or different and is hydrogen or alkyl. Preferably, R.sub.1 is 
a hydrogen or a linear or branched alkyl having 1 to 10 carbons. Exemplary 
alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, isobutyl, 
sec-butyl, tert-butyl, pentyl, isopentyl, neopentyl, tert-pentyl, hexyl, 
dimethylbutyl, heptyl, octyl, nonyl and decyl. R.sub.2 and R.sub.3 are 
hydrogen or linear or branched alkyl having 1 to 4 carbons. Examples of 
such alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, 
isobutyl, sec-butyl and tert-butyl. In a preferred embodiment, R.sub.3 is 
hydrogen. At least one of R.sub.1, R.sub.2 and R.sub.3 must be an alkyl. 
R is hydrogen, alkyl, alkenyl, alkynyl, hydroxyalkynyl, alkoxy, phenyl, 
hydroxy, amino, alkylamino, phenylamino, or halogen. Preferred alkyl 
groups are linear or branched alkyls having 1 to 20 carbons. Exemplary 
alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, isobutyl, 
sec-butyl, tert-butyl, pentyl, isopentyl, neopentyl, tert-pentyl, hexyl, 
dimethylbutyl, heptyl, octyl, nonyl, decyl and stearyl. 
Preferred alkenyl groups are linear or branched alkenyls having 2 to 4 
carbons. Exemplary alkenyl groups are ethenyl, 1-propenyl, 2-propenyl, 
1-butenyl, 2-butenyl and 3-butenyl. 
Preferably the alkynyl is a linear or branched alkynyl having 2 to 20 
carbons. Exemplary alkynyl groups include ethynyl, 1-propynyl, 2-propynyl, 
1-butynyl, 2-butynyl, 3-butynyl, sec-butynyl, pentynyl, isopentynyl, 
2-pentynyl, 3-pentynyl, 4-pentynyl, hexynyl, heptynyl, octynyl, nonynyl, 
decynyl and stearynyl. 
If R is a hydroxyalkynyl, it is preferred that the alkynyl is substituted 
with one or more hydroxy groups. 
Preferred alkoxy groups include linear or branched alkoxy having 1 to 4 
carbons. Examples of such groups include methoxy, ethoxy, propoxy, 
isopropoxy, butoxy, isobutoxy, sec-butoxy, and tert-butoxy. 
Preferred alkylamino groups include linear or branched alkylamino groups 
having 1 to 10 carbons. Examples of such groups include methylamino, 
dimethylamino, ethylamino, diethylamino, methylethylamino, propylamino, 
isopropylamino, butylamino, isobutylamino, sec-butylamino, 
tert-butylamino, pentylamino, isopentylamino, neopentylamino, 
tert-pentylamino, hexylamino, dimethylbutylamino, heptylamino, octylamino, 
nonylamino and decylamino. 
Exemplary halogen groups include fluoro, chloro, bromo, and iodo. 
X is hydrogen, alkyl, alkynyl, allyl, methallyl, cycloalkyl, alkyl having 
one or more hydroxy groups, phenyl, substituted phenyl, alkyl having one 
or more phenyl groups, alkyl having one or more substituted phenyl groups, 
bicycloalkyl, naphthylalkyl, acenaphthylenylalkyl or a compound 
represented by Formula (II) or Formula (III). The substituted phenyl 
groups include a phenyl which has been substituted with one or more 
halogen, lower alkyl, lower alkoxy and/or trifluoromethyl substituents. 
Preferred alkyl groups are linear or branched alkyls having 1 to 6 carbons. 
Exemplary alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, 
isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl, neopentyl, 
tert-pentyl, hexyl and dimethylbutyl. 
Preferred alkynyl groups are linear or branched alkynyls having 2 to 7 
carbons. Examples of such groups include ethynyl, 1-propynyl, 2-propynyl, 
1-butynyl, 2-butynyl, 3-butynyl, sec-butynyl, pentynyl, isopentynyl, 
2-pentynyl, 3-pentynyl, 4-pentynyl, hexynyl and heptynyl. 
Preferred cycloalkyl groups have 3 to 8 carbons. Examples of such groups 
include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and 
cyclooctyl. 
Preferred alkyl groups having one or more hydroxy substituents are linear 
or branched alkyls having 1 to 4 carbons. Examples of such groups include 
methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl and 
tert-butyl substituted with one or two hydroxy groups. 
Preferred alkyl groups having one or more phenyl or substituted phenyl 
substituents are phenylalkyl or diphenylalkyl in which one or two 
substituted phenyl as defined above is/are bonded to a linear or branched 
alkyl having 1 to 3 carbons. Examples of such alkyl groups are methyl, 
ethyl, propyl, and isopropyl. 
Preferred bicycloalkyl groups are endo- or exo-bicyclo 2,2,1!heptyl. 
Preferred naphthylalkyls are those in which the naphthyl group is bonded to 
an alkyl having 1 to 3 carbons. Examples of such alkyl groups are methyl, 
ethyl, propyl and isopropyl. 
Preferred acenaphthylenylalkyls are those in which acenaphthylenyl is 
bonded to an alkyl having 1 to 3 carbons. Examples of such alkyl groups 
are methyl, ethyl, propyl and isopropyl. It should be noted that the 
1,2-dihydro form of an acenaphthylenylalkyl may also be used within the 
context of the instant invention. X may also be a group represented by 
either Formula (II) or Formula (III). 
In a preferred embodiment, X is either hydrogen or an alkyl having 1 to 3 
carbons. 
In the general formulae (II) and (III), Z is hydrogen, hydroxy or lower 
alkoxy. Preferably Z is a linear or branched alkoxy having 1 to 3 carbons. 
Exemplary alkoxy groups are methoxy, ethoxy, propoxy and isopropoxy. Q is 
hydrogen or hydroxy. A is --CH.sub.2 --, --O--, --S-- or a single bond 
forming a five-membered ring; Y is --(CH.sub.2)n-- or a single bond; and n 
is an integer from 1 to 3. In Formula (II) a broken line symbolizes the 
presence of either a single bond or a double bond. 
When A is a single bond forming a five-membered ring the following groups 
result: 
##STR7## 
When Y is a single bond the following groups result: 
##STR8## 
The adenosine derivatives of the present invention include the following 
novel compounds: 
(1) Adenosine derivatives represented by the following general formula 
(Ia): 
##STR9## 
wherein each of R.sub.1 a, R.sub.2 a and Xa may be the same or different 
and is hydrogen or alkyl; 
Ra is an alkyl having more than 6 carbon atoms, alkenyl, alkynyl, 
hydroxyalkynyl, alkoxy, phenyl, hydroxy, alkylamino or phenylamino; and 
at least one of R.sub.1 a and R.sub.2 a is alkyl. 
(2) Adenosine derivatives represented by the following general formula 
(Ib): 
##STR10## 
wherein each of R.sub.1 b and R.sub.2 b may be the same or different and 
is alkyl; 
Rb and Xb is hydrogen or alkyl; and 
at least one of Rb and Xb is an alkyl. 
(3) Adenosine derivatives represented by the following general formula 
(Ic): 
##STR11## 
wherein each of R.sub.1 c, R.sub.2 c and Xc may be the same or different 
and is hydrogen or an alkyl; 
Rc is bromo or iodo; and 
at least one of R.sub.1 c and R.sub.2 c is an alkyl. 
(4) Adenosine derivatives represented by the following general formula 
(Id): 
##STR12## 
wherein R.sub.1 d is an alkyl having more than 2 carbons. 
Preferred substituents in the said adenosine derivatives represented by 
formulae (Ia) to (Id) are the same groups indicated in the formula (I). 
Examples of the compound which is contained as an effective component in 
the adenosine deaminase inhibitor in accordance with the present invention 
are as follows: 
(Compound 1) 2'-O-Methyladenosine; 
(Compound 2) 3'-O-Methyladenosine; 
(Compound 3) 2'-O-Ethyl adenosine; 
(Compound 4) 2'-O-n-Butyladenosine; 
(Compound 5) 2,2'-O-Dimethyl adenosine; 
(Compound 6) 2,3'-O-Dimethyl adenosine; 
(Compound 7) 2-Isopropyl-2'-O-methyladenosine; 
(Compound 8) 2-Isopropyl-3'-O-methyladsnosine; 
(Compound 9) 2-Methoxy-3'-O-methyladsnosine; 
(Compound 10) 2-Methyl-2'-O-ethyl adenosine; 
(Compound 11) 2-Methyl-2'-O-n-butyl adenosine; 
(Compound 12) 5'-O-Methyl adenosine; 
(Compound 13) 5'-O-n-Butyl adenosine; 
(Compound 14) 2',5'-O-Dimethyladenosine; 
(Compound 15) 3',5'-O-Dimethyl adenosine; 
(Compound 16) 2,5'-O-Dimethyl adenosine; 
(Compound 17) 2-Methyl-5'-O-n-butyladenosine; 
(Compound 18) N.sup.6,2'-O-Dimethiladenosine; 
(Compound 19) N.sup.6 -Ethyl-2'-O-methyladenosine; 
(Compound 20) N.sup.6 -n-Butyl-2'-O-methyladenosine; 
(Compound 21) N.sup.6 -Methyl-2'-O-ethyladenosine; 
(Compound 22) N.sup.6 -Methyl-2'-O-n-butyladenosine; 
(Compound 23) N.sup.6 -Methyl-2'-O-n-hexyladenosine; 
(Compound 24) N.sup.6 -Methyl-2'-O-n-octyladenosine; 
(Compound 25) N.sup.6, 5'-O-Dimethyladenosine; 
(Compound 26) N.sup.6 -n-Butyl-5'-O-methyladenosine; 
(Compound 27) 2, N.sup.6,2'-O-Trimethyladenosine; 
(Compound 28) 2, N.sup.6 -Dimethyl-2'-O-ethyladenosine; 
(Compound 29) N.sup.6 -n-Butyl-2,2'-O-dimethyladenosine; 
(Compound 30) N.sup.6,2'-O-Dimethyl-2-hexyladenosine; 
(Compound 31) N.sup.6,2'-O-Dimethyl-2-decyladenosine; 
(Compound 32) N.sup.6,2'-O-Dimethyl-2-(1-hexyn-1-yl)-adenosine; 
(Compound 33) N.sup.6,2'-O-Dimethyl-2-(1-dodecyn-1-yl)-adenosine; 
(Compound 34) 2,N.sup.6,5'-O-Trimethyladenosine; 
(Compound 35) N.sup.6 -n-Butyl-2,5'-O-dimethyladenosine; 
(Compound 36) 2,N.sup.6,3'-O-Trimethyladenosine; 
(Compound 37) 2-Phenyl-2'-O-methyladenosine; 
(Compound 38) 2-Phenyl-3'-O-methyladenosine; 
(Compound 39) 2-Hydroxy-2'-O-methyladenosine; 
(Compound 40) 2-Hydroxy-3'-O-methyladenosine; 
(Compound 41) 2-Chloro-2'-O-methyladenosine; 
(Compound 42) 2-Chloro-3'-O-methyladeonsine; 
(Compound 43) 2-Bromo-2'-O-methyladenosine; 
(Compound 44) 2-Bromo-3'-O-methyladeonsine; 
(Compound 45) 2-Bromo-N.sup.6,2'-O-dimethyladenosine; 
(Compound 46) 2-Bromo-N.sup.6,3'-O-dimethyladeonsine; 
(Compound 47) 2-Iodo-2'-O-methyladenosine; 
(Compound 48) 2-Iodo-3'-O-methyladeonsine; 
(Compound 49) 2-Fluoro-2'-O-methyladenosine; 
(Compound 50) 2-Amino-2'-O-methyladenosine; 
(Compound 51) 2-Amino-3'-O-methyladenosine; 
(Compound 52) 2-Pentylamino-2'-O-methyladenosine; 
(Compound 53) 2-Phenylamino-2'-O-methyladenosine; 
(Compound 54) 2-Phenylamino-3'-O-methyladenosine; 
(Compound 55) 2-Phenylamino-N.sup.6,2'-O-dimethyladenosine; 
(Compound 56) 2-Phenylamino-N.sup.6,3'-O-dimethyladenosine; 
(Compound 57) 2-(3-Hydroxy-1-propyn-1-yl)-2'-O-methyladenosine; 
(Compound 58) 2-(3-Hydroxy-1-propyn-1-yl)-3'-O-methyladenosine; 
(Compound 59) 2',3'-O-Dimethyladenosine; 
(Compound 60) N.sup.6,2',3'-O-Trimethyladenosine; 
(Compound 61) N.sup.6 -Methyl-2',3'-O-diethyladenosine; 
(Compound 62) N.sup.6 -n-Butyl-2',3'-O-dimethyladenosine; 
(Compound 63) 2,2',3'-O-Trimethyladenosine; 
(Compound 64) 2,N.sup.6,2',3'-O-Tetramethyladenosine; 
(Compound 65) N.sup.6 -Allyl-2'-O-methyladenosine; 
(Compound 66) N.sup.6 -Methallyl-2'-O-methyladenosine; 
(Compound 67) N.sup.6 -(2,3-Dihydroxypropyl)-2'-O-methyladenosine; 
(Compound 68) N.sup.6 -Cyclopropyl-2'-O-methyladenosine; 
(Compound 69) N.sup.6 -Cyclopentyl-2'-O-methyladenosine; 
(Compound 70) N.sup.6 -Cyclopentyl-2'-O-ethyladenosine; 
(Compound 71) N.sup.6 -Cyclopentyl-2,2'-dimethyladenosine; 
(Compound 72) N.sup.6 -Cyclopentyl-2-bromo-2'-O-methyladenosine; 
(Compound 73) N.sup.6 -Cyclohexyl-2'-O-methyladenosine; 
(Compound 74) N.sup.6 -Cyclohexyl-2,2'-O-dimethyladenosine; 
(Compound 75) N.sup.6 -Cycloheptyl-2'-O-methyladenosine; 
(Compound 76) N.sup.6 P-Methoxyphenyl-2'-O-methyladenosine; 
(Compound 77) N.sup.6 -P-Fluorophenyl-2'-O-methyladenosine; 
(Compound 78) N.sup.6 P-Chlorophenyl-2'-O-methyladenosine; 
(Compound 79) N.sup.6 Benzyl-2'-O-methyladenosine; 
(Compound 80) N.sup.6 -(R)-Phenyl-isopropyl-2'-O-methyladenosine; 
(Compound 81) N.sup.6 -(2,2-Diphenylethyl)-2'-O-methyladenosine; 
(Compound 82) N.sup.6 -(exo-Dicyclo2,2,1!heptyl)-2'-O-methyl-adenosine; 
(Compound 83) N.sup.6 -(endo-Dicyclo2,2,1!heptyl)-2'-O-methyladenosine; 
(Compound 84) N.sup.6 -(1-Naphthyl)methyl-2'-O-methyladenosine; 
(Compound 85) N.sup.6 -1-(Acenaphthylenyl)methyl-2'-O-methyladenosine; 
(Compound 86) N.sup.6 
-(1,2-Dihydro-1-acenaphthylenyl)methyl-2'-O-methyladenosine; 
(Compound 87) N.sup.6 -(2,3-Dihydro-1H-inden-1-yl)-2'-O-methyladenosine; 
(Compound 88) N.sup.6 -(2,3-Dihydro-1H-inden-2-yl)-2'-O-methyladenosine; 
(Compound 89) N.sup.6 
-(2,3-Dihydro-1H-inden-1-yl)methyl-2'-O-methyladenosine; 
(Compound 90) N.sup.6 -(3H-Inden-1-yl)methyl-2'-O-methyladenosine; 
(Compound 91) N.sup.6 
-(5-Methoxy-2,3-dihydro-1H-inden-2-yl)-2'-O-methyladenosine; 
(Compound 92) N.sup.6 -(1-Tertrahydronaphtyl)-2'-O-methyladenosine; 
(Compound 93) N.sup.6 -(2-Tertrahydronaphtyl)-2'-O-methyladenosine; 
(Compound 94) N.sup.6 -(3,4-Dihydro-1-naphthyl)methyl-2'-O-methyladenosine; 
(Compound 95) N.sup.6 
-(5-Hydroxy-1-tetrahydronaphthyl)-2'-O-methyladenosine; 
(Compound 96) N.sup.6 
-(1-Hydroxy-1-tetrahydronaphthyl)-methyl-2'-O-methyladenosine; 
(Compound 97) N.sup.6 
-(5-Methoxy-1-tetrahydronaphthyl)-2'-O-methyladenosine; 
(Compound 98) N.sup.6 
-(6-Methoxy-1-tetrahydronaphthyl)-2'-O-methyladenosine; 
(Compound 99) N.sup.6 
-(7-Methoxy-1-tetrahydronaphthyl)-2'-O-methyladenosine; 
(Compound 100) N.sup.6 -(4-Chromanyl)-2'-O-methyladenosine; 
(Compound 101) N.sup.6 -(4-Thiochromanyl)-2'-O-methyladenosine; 
(Compound 102) N.sup.6 -Fluorenyl-2'-O-methyladenosine; 
(Compound 103) N.sup.6 -(9-Fluorenyl)methyl-2'-O-methyladenosine; 
(Compound 104) N.sup.6 -(9-Hydroxy-9-fluorenyl)methyl-2'-O-methyladenosine; 
and 
(Compound 105) N.sup.6 -(9-Xanthenyl)methyl-2'-O-methyladenosine. 
The compounds of the present invention as given above may be prepared by a 
method disclosed, for example, in U.S. Pat. No. 4,843,066 and 
corresponding Japanese Laid Open (Kokai) No. 63/239,294, U.S. Pat. No. 
4,985,409 and corresponding Japanese Laid Open (Kokai) No. 02/184,696, and 
Great Britain Patent No. 2,226,027A and corresponding Japanese Laid Open 
(Kokai) No. 02/218,689. 
For example, the adenosine derivatives of the present invention can be 
prepared as follows: 
(1) Adenosine or adenosine derivatives having a lower alkyl group, an amino 
group or halogen at the 2-position may be alkylated at the 2'-O- or 
3'-O-position by an alkylating agent to give the compounds of the present 
invention. A diazoparaffin, such as diazomethane, diazoethane, 
diazopropane or diazobutane, can be used as the alkylating agent. The 
appropriate solvent which does not inhibit the reaction such as 
1,2-dimethoxyethane can be preferably used. This O-alkylating reaction can 
be carried out as follows: (i) The reaction mixture is reacted for several 
minutes to several hours at room temperature in the presence of a catalyst 
such as p-toluenesulfonic acid; (ii) The starting material is dissolved in 
about 80.degree. C. hot water and the alkylating agent such as 
diazoparaffin is added thereto, and the reaction mixture is reacted for 
several hours to a day. 
(2) Both 3'-O- and 5'-O-positions of the adenosine derivatives are 
protected by tetraisopropyldisiloxane (TIPDS) group to carry out 
O-alkylation selectively at the 2'-O-position. A 6-chloropurine-9-riboside 
and TIPDSCL.sub.2 (1,3-dichloro-1,1,3,3-tetraisopropyldisiloxane 
dichloride) are stirred for several hours at room temperature to protect 
the 3'-O- and 5'-O-positions, and then the 2'-O-position of the compound 
protected by TIPDS can be selectively alkylated by an alkylating agent 
such as methyl iodide, ethyl iodide, propyl iodide or butyl iodide in the 
presence of a catalyst such as silver oxide. After the 2'-O-alkylation, an 
amination or alkylamination at the 6-position can be carried out by 
reacting with ammonia or an alkylamine such as methylamine, ethylamine, 
propylamine or butylamine with heating. The protecting group, TIPDS, can 
be removed by a conventional method to give the compounds of the present 
invention. 
(3) In the similar manner, both 2'-O- and 3'-O-positions of the adenosine 
derivatives are protected by isopropylidene group to carry out 
O-alkylation selectively at the 5'-O-position. Namely, a 
6-chloropurine-9-riboside and 2,2-dimethoxypropane are reacted for several 
hours at room temperature in the presence of a catalyst such as 
p-toluenesulfonic acid to carry out isopropylidenation. After the 
5'-O-alkylation, an amination or alkylamination at 6-position can be 
carried out as mentioned above. The protecting group, isopropylidene 
group, can be removed by a conventional method, for example, treatment 
with formic acid, to give the compounds of the present invention. 
The resulting compounds of the present invention can be purified by known 
methods such as distillation chromatography and recrystallization. 
Identification is established through, inter alia, melting point, 
elemental analysis, IR, NMR, UV, mass spectrum, etc. 
Adenosine derivatives of the present invention include the pharmaceutically 
acceptable salts of the compounds represented by the general formula. 
Examples of such salts are acid addition salts such as salts of 
hydrochloric acid, sulfuric acid, nitric acid, hydrobromic acid, 
phosphoric acid, perchloric acid, thiocyanic acid, boric acid, formic 
acid, acetic acid, haloacetic acid, propionic acid, glycolic acid, citric 
acid, tartaric acid, succinic acid, gluconic acid, lactic acid, malonic 
acid, fumaric acid, anthranilic acid, benzoic acid, cinnamic acid, 
p-toluenesulfonic acid, naphthalenesulfonic acid and sulfanilic acid and 
salts with alkali metal (e.g. sodium and potassium), alkali earth metal 
(e.g. calcium and magnesium) and metal (e.g. aluminum). 
Adenosine derivatives of the present invention include metal complexes 
thereof such as, for example, complexes with zinc, nickel, cobalt, copper 
and iron. 
These salts and metal complexes may be manufactured from free adenosine 
derivatives of the present invention or may be transformed each other. 
When there are stereoisomers for the compounds of the present invention 
such as cis-trans isomers, optical isomers and conformational isomers, the 
present invention includes all of them.

The following descriptions serve to illustrative examples for preparation 
of the compounds of the present invention: 
EXAMPLE 1 
10 g of 6-chloro-9-(3,5-O-TIPDS)-.beta.-D-ribofuranosyl-9H-purine was 
dissolved in 50 ml of ethyl iodide, and silver oxide was added and stirred 
with heating. The reaction mixture was applied on silica gel column, 
washed with benzene and then eluted with ethyl acetate/hexane. The eluate 
was concentrated to dryness under reduced pressure. The residue was 
dissolved in benzene and a 40% (W/V) aqueous solution of monomethylamine 
was added thereto. After stirring overnight, the benzene layer was 
separated, washed with 1N HCl and brine, and a mixture of 1M 
tetra-n-butylammonium and tetrahydrofuran was added thereto. The reaction 
mixture was stirred for 30 minutes at room temperature concentration under 
reduced pressure, and then purified by silica gel column to give 2.1 g of 
N.sup.6 -methyl-2'-O-ethyladenosine (Compound 21). 
.sup.1 H-NMR (D.sub.2 O): 1.21(3H,t), 3.19(3H,s), 3.58(1H,m), 3.69(1H,m), 
3.85(1H,dd), 3.93(1H,dd), 4.32(1H,m), 4.55(1H,dd), 4.63(1H,dd), 
6.10(1H,dd), 8.25(1H,s), 8.30(1H,s) 
In the same manner as mentioned above, the following compounds were 
obtained. 
N.sup.6 -Methyl-2'-O-n-butyladenosine (Compound 22) 
.sup.1 H-NMR (DMSO-d.sub.6): 0.74(3H,t), 1.17(2H,m), 1.37(2H,m), 
2.96(3H,s), 3.34(1H,m), 3.55(2H,m), 3.68(1H,m), 3.99(1H,m), 4.29(1H,m), 
4.47(1H,m), 5.15(1H,d), 5.42(1H,m), 5.99(1H,d), 7.81(1H,s), 8.23(1H,s), 
8.37(1H,s) 
N.sup.6 -Methyl-2'-O-n-hexyladenosine (Compound 23) 
.sup.1 H-NMR (CDCl.sub.3): 0.86(3H,t), 1.16(8H,m), 1.40(2H,m), 3.21(3H,s), 
3.34(1H,m), 3.48(1H,m), 3.75(1H,m), 3.97(1H,m), 4.36(1H,m), 4.53(1H,d), 
4.82(1H,dd), 5.82(1H,d), 5.87(1H,s), 6.87(1H,dd), 7.76(1H,s), 8.37(1H,s) 
N.sup.6 -Methyl-2'-O-n-octyladenosine (Compound 24) 
.sup.1 H-NMR (CDCl.sub.3): 0.86(3H,t), 1.16(8H,m), 1.24(2H,m), 1.41(2H,m), 
3.21(3H,s), 3.33(1H,m), 3.49(1H,m), 3.76(1H,m), 3.97(1H,m), 4.36(1H,m), 
4.53(1H,d), 4.82(1H ,dd), 5.82(1H,d), 6.02(1H,s), 6.87(1H,dd), 7.77(1H,s), 
8.37(1H,s) 
EXAMPLE 2 
5.34 g of adenosine was dissolved in 80 ml of dimethylformamide and 800 mg 
of 60% W/W) sodium hydride in mineral oil was added thereto. After 
stirring for 30 minutes in ice-cold water, 2.84 g of methyl iodide in 10 
ml of dimethylformamide was added dropwise. The reaction mixture was 
stirred under cooling for 2 hours and concentrated to dryness under 
reduced pressure. The residue was dissolved in water and applied on cation 
exchange column. The fraction was then eluted with 10% (V/V) aqueous 
solution of methanol collected, concentrated to dryness, and purified by 
silica gel column to give 1.36 g of 2',3'-O-dimethyladenosine. 
m.p.: 180.degree. C. 
.sup.1 H-NMR (DMSO-d.sub.6): 3.32(3H,s), 3.40(3H,s), 3.57(1H,m), 
3.70(1H,m), 4.09(2H,m), 4.54(1H,dd), 5.47(1H,dd), 6.00(1H,d), 
7.35(2H,s,D.sub.2 O-Disappearance), 8.14(1H,s), 8.38(1H,s) 
EXAMPLE 3 
2 g of the compound obtained in Example 2 and 2 ml of methyl iodide were 
dissolved in dimethylacetoamide, and stirred overnight at room 
temperature. The reaction mixture was concentrated to dryness under 
reduced pressure and 10 ml of 2N sodium hydroxide solution was added 
thereto. The solution was refluxed for one hour with heating. After 
cooling to room temperature, the solution was neutralized with 2N HCl and 
applied on Amberlite XAD-7 column. The column was washed with water and 
eluted with 50% (V/V) aqueous solution of methanol. The eluate was 
concentrated to dryness under reduced pressure and recrystallized from 
ethyl acetate to give 1.8 g of N.sup.6,2',3'-O-trimethyladenosine 
(Compound 60). 
m.p.: 165.degree. C. 
.sup.1 H-NMR (DMSO-d.sub.6): 2.96(3H,s), 3.31(3H,s), 3.40(3H,s), 
3.57(1H,m), 3.70(1H,m), 4.09(2H,m), 4.54(1H,dd), 5.49(1H,dd), 6.01(1H,d), 
7.83 (1H,s,D.sub.2 O-Disappearance), 8.24(1H,s), 8.38(1H,s) 
EXAMPLE 4 
14.3 g of 6-chloro-9-.beta.-D-ribofuranosyl-9H-purine and 15 g of 
triphenylchlorosilane were dissolved in 500 ml of pyridine and stirred for 
one hour at room temperature. Pyridine was distilled away and the residue 
was dissolved in benzene. The benzene layer was washed with 1N HCl and 
brine, and then dried over sodium sulfate anhydride. The solvent was 
distilled off and the residue was recrystallized from a mixture of hexane 
and ethyl acetate to give 21.5 g of 
6-chloro-9-15-O-triphenylsilyl-.beta.-D-ribofuranosyl)-9H-purine. 
5.45 g of the resulting product was dissolved in 50 ml of ethyl iodide and 
silver oxide was added thereto with heating. The reaction mixture was 
applied on silica gel column. The column was washed with benzene and 
eluted with a mixture of hexane and ethyl acetate. The eluate was 
concentrated to dryness under reduced pressure. The residue was dissolved 
in benzene and a 40% (W/V) aqueous solution of monomethylamine was added 
thereto. After stirring overnight, the benzene layer was collected, washed 
with 1N HCl and brine and purified by silica gel column to give 
6-amino-9-(5-O-triphenylsilyl-.beta.-D-ribofuranosyl)-9H-purine. The 
resulting product was dissolved in tetrahydrofuran and 
tetra-n-butylammonium floride in tetrahydrofurane was added thereto. After 
stirring for 30 minutes at room temperature, the solution was concentrated 
to dryness under reduced pressure and purified by silica gel column to 
give 600 mg of N.sup.6 -methyl-2',3'-O-diethyladenosine (Compound 61). 
m.p.: amorphous 
.sup.1 H-NMR (MeOH-d.sub.4): 1.07(3H,t), 1.22(3H,t), 3.08(3H,s), 
3.48(1H,m), 3.58(1H,m), 3.65(2H,m), 3.70(1H,dd), 3.87(2H,m), 4.20(1H,dd), 
6.00(1H,d), 8.20(1H,s), 8.23(1H,s) 
EXAMPLE 5 
2-Methyladenosine was dialkylated in the same manner as Example 2 to give 
2,2',3'-O-trimethyladenosine (Compound 63). 
m.p.: 156.degree. C. 
.sup.1 H-NMR (DMSO-d.sub.6): 2.39(3H,s), 3.28(3H,s), 3.40(3H,s), 
3.58(1H,m), 3.70(1H,m), 4.08(1H,m), 4.13(1H,m), 4.52(1H,m), 
5.80(1H,m,D.sub.2 O-Disappearance),5.95(1H,d), 7.27(2H,s,D.sub.2 
O-Disappearance), 8.28(1H,s) 
EXAMPLE 6 
Compound 63 was methylated by methyl iodide and then rearranged to give 
2,N.sup.6,2',3'-O-tetramethyladenosine (Compound 64). 
.sup.1 H-NMR (DMSO-d.sub.6): 2.43(3H,s), 2.94(3H,s), 3.28(3H,s), 
3.40(3H,s), 3.57(1H,m), 3.70(1H,m), 4.08(1H,m), 4.13(1H,m), 4.52(1H,m), 
5.79(1H,m,D.sub.2 O-Disappearance), 5.96(1H,d), 7.71(1H,s,D.sub.2 
O-Disappearance), 8.26(1H,s) 
EXAMPLE 7 
1.2 g of 2-iodoadenosine was suspended in 150 ml of 1 mmol tin chloride 
dihydrate/methanol. 50 ml of 0.4-0.5M diazomethane in 1,2-dimethoxyethane 
was added with stirring. After stirring for one hour at room temperature, 
the reaction mixture was concentrated to dryness under reduced pressure. 
The resulting product was applied on ODS column and eluted with 40% (V/V) 
methanol in 0.1% (V/V) aqueous solution of TFA. First, 
2-iodo-2'-O-methyladenosine (Compound 47) was eluted, and then 
2-iodo-3'-O-methyladenosine (Compound 48) was eluted. Both fractions were 
concentrated to dryness to give 135 mg of Compound 47 and 56 mg of 
Compound 48. 
2-iodo-2'-O-methyladenosine (Compound 47) 
.sup.1 H-NMR (D.sub.2 O): 3.52(3H,s), 3.92(1H,dd), 3.97(1H,dd), 4.27(1H,m), 
4.54(1H,dd), 4.61 (1H,dd), 6.02(1H,d), 7.98(1H,s) 
2-iodo-3'-O-methyladenosine (Compound 48) 
.sup.1 H-NMR (D.sub.2 O): 3.58(3H,s) 3.86(1H,dd) 3.94(1H,dd), 4.18(1H,dd) 
4.35(1H,m) 4.97(1H,dd) 5.93(1H,d), 7.94(1H,s) 
In the same manner the following compounds were obtained by using 
2-substitutedadenosine, N.sup.6 -substitutedadenosine or 2, N.sup.6 
-disubstitutedadenosine. 
2-Methoxy-3'-O-methyladenosine (Compound 9) 
.sup.1 H-NMR (D.sub.2 O): 3.56(3H,s) 3.84(1H,dd), 3.94(1H,dd), 3.96(3H,s) 
4.22(1H,dd) 4.33(1H,m) 5.06(1H,dd), 5.98(1H,d) 8.10(1H,s,8-H) 
N.sup.6,2'-O-Dimethyl-2-hexyladenosine (Compound 30) 
.sup.1 H-NMR (MeOH-d.sub.4): 0.88(3H,t) 1.33(6H,m), 1.78(2H,quintet), 
2.74(2H,t), 3.12(3H,brs), 3.37(3H,s), 3.73(1H,dd), 3.88(1H,dd), 
4.18(1H,m), 4.49(2H,m), 5.98(1H,d), 8.10(1H,s,8-H) 
N.sup.6,2'-O-Dimethyl-2-decyladenosine (Compound 31) 
.sup.1 H-NMR (MeOH-d.sub.4): 0.88(3H,t), 1.27-1.33(14H,m), 
1.78(2H,quintet), 2.75(2H,t), 3.13(3H,brs), 3.37(3H,s), 3.73(1H,dd), 
3.89(1H,dd), 4.18(1H,m), 4.49(2H,m) 
N.sup.6,2'-O-Dimethyl-2-(1-hexyn-1-yl)adenosine (Compound 32) 
.sup.1 H-NMR (MeOH-d.sub.4): 0.97(3H,t), 1.52(2H,m), 1.62(2H,m), 
2.54(2H,t), 3.09(3H,brs), 3.40(3H,s), 3.74(1H,dd), 3.88(1H,dd), 
4.15(1H,m), 4.42(1H,dd), 4.48(1H,dd), 6.00(1H,d), 8.22(1H,s) 
N.sup.6,2'-O-Dimethyl-2-(1-dodecyl-1-yl)adenosine (Compound 33) 
.sup.1 H-NMR (MeOH-d.sub.4): 0.89(3H,t), 1.30(14H,m), 1.49(2H,m), 
1.63(2H,quintet), 2.43(2H,t), 3.41(3H,s), 3.74(1H,dd), 3.88(1H,dd), 
4.15(1H,m), 4.41(1H,dd), 4.48(1H,dd), 6.00(1H,d), 8.30(1H,s) 
2-Phenyl-2'-O-methyladenosine (Compound 37) 
.sup.1 H-NMR (MeOH-d.sub.4): 3.53(3H,s), 3.77(1H,dd), 3.91(1H,dd), 
4.12(1H,m), 4.50(1H,dd), 4.54(1H,dd), 6.20(1H,d), 7.43(3H,m), 8.32(2H,m), 
8.36(1H,s) 
2-Phenyl-3'-O-methyladenosine (Compound 38) 
.sup.1 H-NMR (MeOH-d.sub.4): 3.53(3H,s), 3.74(1H,dd), 3.88(1H,dd), 
4.10(1H,dd), 4.22(1H,m), 5.00(1H,dd), 6.07(1H,d), 7.42(3H,m), 8.32(3H,m) 
2-Bromo-2'-O-methyladenosine (Compound 43) 
.sup.1 H-NMR (D.sub.2 O): 3.50(3H,s), 3.91(1H,dd), 3.99(1H,dd), 4.37(1H,m), 
4.57(1H,dd), 4.67(1H,dd), 6.13(1H,d), 8.37(1H,s) 
2-Bromo-3'-O-methyladenosine (Compound 44) 
.sup.1 H-NMR (D.sub.2 O): 3.60(3H,s), 3.90(1H,dd), 4.01(1H,dd), 
4.19(1H,dd), 4.44(1H,m), 4.95(1H,dd), 6.07(1H,d), 8.37(1H,s) 
2-Bromo-N.sup.6,2'-O-dimethyladenosine (Compound 45) 
.sup.1 H-NMR (D.sub.2 O): 3.22(3H,s), 3.57(3H,s), 3.90(1H,dd), 3.98(1H,dd), 
4.29(1H,m), 4.59(1H,dd), 4.65(1H,dd), 6.12(1H,d), 8.16(1H,s) 
2-Bromo-N.sup.6,3'-O-dimethyladenosine (Compound 46) 
.sup.1 H-NMR (D.sub.2 O): 3.07(3H,s), 3.60(3H,s), 3.93(1H,dd), 3.99(1H,dd), 
4.23(1H,dd), 4.36(1H,m), 5.03(1H,dd), 6.03(1H,d), 8.14(1H,s) 
2-Pentylamino-2'-O-methyladenosine (Compound 52) 
.sup.1 H-NMR (MeOH-d.sub.4): 0.94(3H,t), 1.38(4H,m), 1.65(2H,m), 
3.42(2H,t), 3.49(3H,s), 3.76(1H,dd), 3.87(1H,dd), 4.08(1H,m), 4.25(1H,t), 
4.43(1H,t), 5.99(1H,d), 8.22(1H,s) 
2-Phenylamino-2'-O-methyladenosine (Compound 53) 
.sup.1 H-NMR (MeOH-d.sub.4): 3.46(3H,s), 3.74(1H,dd), 3.85(1H,dd), 
4.07(1H,m), 4.23(1H,dd), 4.39(1H,dd), 6.03(1H,d), 7.11(1H,t), 7.35(2H,t), 
7.60(2H,d), 8.35(1H,s) 
2-Phenylamino-3'-O-methyladenosine (Compound 54) 
.sup.1 H-NMR (MeOH-d.sub.4): 3.46(3H,s), 3.69(1H,dd), 3.79(1H,dd), 
3.94(1H,dd), 4.16(1H,m), 4.74(1H,dd), 5.91(1H,d), 7.13(1H,t), 7.36(2H,t), 
7.60(2H,d), 8.28(1H,s) 
2-Phenylamino-N.sup.6,2'-O-dimethyladenosine (Compound 55) 
.sup.1 H-NMR (MeOH-d.sub.4): 3.16(3H,s), 3.47(3H,s), 3.77(1H,dd), 
3.88(1H,dd), 4.11(1H,m), 4.20(1H,dd), 4.40(1H,dd), 6.06(1H,d), 7.12(1H,t), 
7.35(2H,t), 7.63(2H,d), 8.42(1H,s) 
2-Phenylamino-N.sup.6,3'-O-dimethyladenosine (Compound 56) 
.sup.1 H-NMR (MeOH-d.sub.4): 3.10(3H,s), 3.49(3H,s), 3.72(1H,dd), 
3.85(1H,dd), 3.98(1H,dd), 4.17(1H,m), 4.85(1H,dd), 5.88(1H,d), 6.94(1H,t), 
7.26(2H,t), 7.69(2H,d), 7.99(1H,s) 
2-(3-Hydroxy-1-propyn-1-yl)-2'-O-methyladenosine (Compound 57) 
.sup.1 H-NMR (D.sub.2 O): 3.45(3H,s), 3.86(1H,dd), 3.95(1H,dd), 4.32(1H,m), 
4.51 (1H,dd), 4.52(2H,s), 4.62(1H,dd), 6.08(1H,d), 8.30(1H,s) 
2-(3-Hydroxy-1-propyn-1-yl)-3'-O-methyladenosine (Compound 58) 
.sup.1 H-NMR (D.sub.2 O+MeOH-d.sub.4): 3.51(3H,s), 3.80(1H,dd), 
3.92(1H,dd), 4.09(1H,m), 4.36(1H,m), 4.47(2H,s), 4.83(1H,dd), 5.96(1H,d), 
8.30(1H,s) 
EXAMPLE 8 
2.2 g of AICA-2'-O-methylriboside was dissolved in 20 ml of 
dimethylformamide and 1.7 g of benzoylisothiocyanate was added thereto. 
After stirring for 3 hours at room temperature, 3.3 g of 
dicyclohexylcarbodiimide was added and reacted for 20 hours at room 
temperature. The reaction mixture was concentrated to an oily residue. To 
the residue, 50 ml of ethanol and 50 ml of aqua ammonia were added, and 
stirred for 18 hours at room temperature. 
The resulting precipitate was collected by filtration to give 1.7 g of 
2-hydroxy-2'-O-methyladenosine (Compound 39). 
.sup.1 H-NMR (D.sub.2 O): 3.46(3H,s), 3.82(1H,dd), 3.90(1H,dd), 4.27(1H,m), 
4.50(1H,dd), 4.59(1H,dd), 5.95(1H,d), 8.03(1H,s) 
In the same manner, AICA-3'-O-methylriboside was used as a starting 
material to give 2-hydroxy-3'-O-methyladenosine (Compound 40). 
.sup.1 H-NMR (D.sub.2 O): 3.54(3H,s), 3.82(1H,dd), 3.94(1H,dd), 
4.10(1H,dd), 4.36(1H,m), 4.86(1H,dd), 5.89(1H,dd), 8.03(1H,s) 
The following descriptions serve to illustrate pharmaceutical studies of 
the compounds of the present invention. 
1. Adenosine Deaminase Inhibiting Action 
The enzymatic reaction was conducted at 25.degree. C. in a 0.05M phosphate 
buffer (pH=7.5). Thus, a reaction solution comprising 800 .mu.l of 
substrate solution (adenosine), 100 .mu.l of a solution to be tested and 
100 .mu.l of enzyme solution (adenosine deaminase of type VII; mucous 
membrane of intestinal tract of calf) was reacted for 5 minutes and the 
reaction was stopped by adding 100 .mu.l of acetic acid. Then the amount 
of inosine produced was determined by means of an HPLC to measure the 
inhibiting activity. The test was conducted for various substrate 
concentrations and Ki values were measured by Lineweaver-Burk plots. 
Examples of the results are given in Table 1. 
TABLE 1 
______________________________________ 
Compound Ki Value 
Tested (M) 
______________________________________ 
1 1.31 .times. 10.sup.-5 
2 4.93 .times. 10.sup.-4 
4 3.47 .times. 10.sup.-6 
5 2.46 .times. 10.sup.-5 
6 1.79 .times. 10.sup.-4 
18 1.24 .times. 10.sup.-6 
19 7.78 .times. 10.sup.-5 
20 2.69 .times. 10.sup.-4 
21 3.30 .times. 10.sup.-7 
22 5.58 .times. 10.sup.-8 
23 8.90 .times. 10.sup.-7 
24 1.10 .times. 10.sup.-7 
25 3.59 .times. 10.sup.-4 
27 8.59 .times. 10.sup.-7 
30 9.00 .times. 10.sup.-8 
31 2.80 .times. 10.sup.-7 
32 1.20 .times. 10.sup.-7 
33 1.50 .times. 10.sup.-5 
34 3.59 .times. 10.sup.-4 
37 6.60 .times. 10.sup.-7 
38 7.90 .times. 10.sup.-6 
39 1.90 .times. 10.sup.-5 
43 3.20 .times. 10.sup.-6 
44 2.20 .times. 10.sup.-5 
45 1.20 .times. 10.sup.-7 
46 3.70 .times. 10.sup.-6 
47 2.30 .times. 10.sup.-7 
48 4.40 .times. 10.sup.-6 
50 2.80 .times. 10.sup.-5 
51 5.00 .times. 10.sup.-6 
52 6.00 .times. 10.sup.-7 
53 1.70 .times. 10.sup.-7 
54 2.20 .times. 10.sup.-6 
55 1.30 .times. 10.sup.-8 
56 1.10 .times. 10.sup.-7 
57 3.00 .times. 10.sup.-5 
59 7.50 .times. 10.sup.-6 
60 7.50 .times. 10.sup.-7 
61 1.50 .times. 10.sup.-7 
63 1.30 .times. 10.sup.-6 
64 8.80 .times. 10.sup.-7 
73 1.03 .times. 10.sup.-3 
______________________________________ 
2. Therapeutic Action to Nephritis. 
When puromycin aminonucleoside is administered to rats, symptoms similar to 
protein-rich urine, hypoproteinemia, hyperlipemia, nephrotic syndrome, 
etc, result and, therefore, rats which are administered with puromycin 
aminonucleoside have been used as pathological model animals for 
nephritis. The chemical name for puromycin aminonucleoside is 
3'-amino-3'-deoxy-N,N-dimethyladenosine. A method by Endo, et al. Sogo 
Rinsho, vol. 38, no. 5, page 821 (1989)! was somewhat modified and used as 
a test method here. Thus, a solution of puromycin aminonucleoside was 
dissolved in a physiological saline liquid and administered just once to a 
tail vein of a male rat (SD strain) of about 200 g body weight at a dose 
of 100 mg/kg (the initial of zero-th day). 
The compound to be tested was dissolved in a physiological saline liquid 
and was given orally for five consecutive days from the zero-th day at a 
dose of 50 mg/kg each. After 24 hours, the accumulated urine was collected 
and the amount of urine and the amount of protein in the urine was 
measured. Blood was collected on the tenth day and the total protein in 
serum, creatinine in serum and urea nitrogen were measured. 
Examples of the results are given in Tables 2 and where the control is 
those animals injected only with the puromycin aminonucleoside: 
TABLE 2 
______________________________________ 
Total Protein 
Serum 
Compound to be 
Urea Nitrogen 
in Serum Creatinine 
Tested (mg/dl) (g/dl) (mg/dl) 
______________________________________ 
Normal 17.6 7.90 0.38 
Control 40.0 7.08 0.85 
Compound 27 31.9 7.44 0.71 
______________________________________ 
TABLE 3 
______________________________________ 
Amount of Protein in Urine 
(mg/day) 
Days Control Compound 27 
______________________________________ 
1st Day 0 0 
2nd 32.0 29.2 
3rd 44.7 33.8 
4th 173.4 122.5 
5th 522.5 399.8 
6th 704.1 443.7 
7th 826.5 558.7 
8th 811.6 595.6 
9th 814.0 593.5 
10th 684.0 528.6 
______________________________________ 
3. Inhibitory Action against Activated Oxygen Generation 
Human peripheral polymorphonuclear leukocyte (PML, 2.times.10.sup.6 cells) 
prepared in a conventional method, bovine heart cytochrome C type-III (75 
nmol), cytocharasin (5 .mu.g) and tested drug were mixed with 
HEPES-buffered saline solution (final 1 ml) and incubated for 5 minutes at 
37.degree. C. N-Formyl-methionyl-leucyl-phenylalanine (FMLP) was added 
(final 10.sup.-7 M) and incubated for 5 minutes. Immediately after the 
incubation, the reaction mixture was centrifuged at 4.degree. C., and then 
the absorbance at 550 nm of the supernatant was measured with a 
spectrophotometer. 
An excess amount of bovine liver superoxide dismutase (SOD) was added to 
the reaction mixture and the absorbance of the supernatant was also 
measured in the same manner as above as a blank value. 
A portion of adenosine deaminase (ADA) may be removed during the 
preparation of human peripheral PML. Therefore, the inhibitory action 
against the generation of superoxide of the tested drug was measured in 
the same manner as mentioned above, adding 0.02 units of bovine liver ADA 
with human peripheral PML. 
The inhibition against superoxide generation was calculated by the 
following equation, and examples of the results are given in Table 4. 
##EQU1## 
TABLE 4 
______________________________________ 
Without ADA With ADA 
Compound 27 (M) 
Inhibition Compound 27 (M) 
Inhibition 
______________________________________ 
1 .times. 10.sup.-3 
100% 1 .times. 10.sup.-4 
88% 
1 .times. 10.sup.-4 
79% 1 .times. 10.sup.-5 
77% 
1 .times. 10.sup.-5 
22% 1 .times. 10.sup.-6 
34% 
1 .times. 10.sup.-6 
2% 1 .times. 10.sup.-7 
10% 
______________________________________ 
4. Suppressive Action against Ischemic Edema 
The right hind paw of ICR-strain male mice (11 weeks of age) were fastened 
with a rubber band to stop the blood stream for 20 minutes, and then the 
rubber band was removed to recover the blood stream. The tested drug was 
administered intravenously before the treatment. Each of the right and 
left hind paws were weighed and the suppressive action was measured 
according to the weight difference between the treated and untreated paws. 
Examples of the results are given in Table 5. 
TABLE 5 
______________________________________ 
Tested Drug Paw Weight (mg) Inhibition 
(10 mg/kg) Treated Untreated (%) 
______________________________________ 
Control 314.8 .+-. 6.8 
219.9 .+-. 6.2 
-- 
Compound 27 
254.4 .+-. 6.6 
223.86.2 67.7 
Allopurinol 
283.2 .+-. 6.3 
222.9 .+-. 6.9 
36.4 
______________________________________ 
5. Effect on the Concentrations of Adenosine and Inosine 
Human peripheral polymorphonuclear leukocyte (2.times.10.sup.6 cells), 
cytochalasin (5 .mu.g) and the tested drug were mixed with HEPES-buffered 
saline solution (final 0.5 ml) and incubated for 5 minutes. FMLP was added 
(final 10.sup.-7 M) and incubated for 10 minutes. As a result of HPLC 
analyses of the reaction mixture, in FMLP-treated group, the inosine peak 
increased compared to the control group. The compound of the present 
invention was added to the FMLP-treated group, which showed a decrease in 
the inosine peak and a significant increase in the adenosine peak compared 
with the group when the compound of the present invention was not added. 
As shown in Table 1, the compounds of the present invention exhibit 
excellent adenosine deaminase inhibiting action. Adenosine deaminase which 
is a metabolic enzyme for adenosine is inhibited whereby the adenosine 
concentration in tissues increases. Neutrophils produce activated oxygen 
when the tissue is in an ischemic state. Adenosine inhibits the production 
of activated oxygen and, in addition, adenosine directly eliminates the 
produced activated oxygen. Further, adenosine lowers the inosine 
concentration whereby it decreases the supply of hypoxanthine. 
Hypoxanthine is a substrate of xanthine-xanthineoxidase system. The 
xanthine-xanthineoxidase system is one of the systems producing the 
activated oxygen. Adenosine deaminase inhibiting substance having a 
production inhibiting action and an eliminating action for activated 
oxygen source as such shows pharmacological actions such as improvement of 
coronary and cerebral blood vessel circulation, prevention and therapy of 
renal diseases, antiinflammatory activity, etc. 
Further, as shown in Tables 2 and 3, the compounds of the present invention 
having adenosine deaminase inhibiting action were evaluated by means of 
pharmacological experiments. Rats which had been administered with 
puromycin aminonucleoside were used as pathological model animals for 
nephritis. Indexes such as total protein in serum, creatinine in serum and 
urea nitrogen concentrations were used to evaluate the therapeutic effects 
of the instant compounds. 
Consequently, the compounds of the present invention having adenosine 
deaminase inhibiting action are useful as pharmaceuticals for the 
prevention and therapy of various kinds of diseases such as ischemic heart 
diseases, diseases caused by cerebrovascular disorder, renal diseases, and 
allergic diseases. Examples of ischemic heart diseases which may be 
treated include angina pectoris, myocardial infarction and arrhythmia. 
Exemplary of diseases caused by cerebrovascular disorder which may be 
treated are cerebral hemorrhage, cerebral infarction, cerebral apoplexy 
and cerebral arteriosclerosis. Nephritis and renal failure are examples of 
renal diseases which may be treated and examples of allergic diseases 
which may be treated include asthma, allergic rhinitis, allergic 
conjunctivitis, urticaris and rheumatism. Moreover, the compounds of the 
present invention are very useful as pharmaceuticals for the prevention 
and therapy of post-operative complicated diseases because they inactivate 
activated oxygen which is generated in ischemic areas during the 
recirculation of blood after operations. 
Adenosine analogs such as 3'-deoxyadenosine and xylosyladenine (anticancer 
drugs) and arabinosyladenine (exhibiting antiherpes activity) are easily 
deaminated by adenosine deaminase in vivo and are inactivated. 
Accordingly, when the compounds of the present invention having adenosine 
deaminase inhibiting action are administered before or together with 
administration of the above-mentioned anticancer drugs or antiviral drugs, 
an effect of inhibiting the decrease in action of such adenosine analogous 
anticancer and antiviral drugs can be expected as well. For the purposes 
of this invention, an adenosine analogous drug is defined as a drug which 
is metabolized or deaminated by adenosine deaminase. 
Adenosine has many pharmacological activities such as cardiovasodilating or 
platelet-aggregation inhibiting activity, so adenosine is used to improve 
blood circulation and treat heart failure, myocardial infarction and other 
such conditions. Adenosine is metabolized by adenosine deaminase and is 
consequently inactive. Accordingly, when the compounds of the present 
invention are administered before or together with the administration of 
adenosine, the instant compounds may inhibit the decrease in such action 
of adenosine. 
The compounds of the present invention can be made into pharmaceuticals by 
combining them with suitable carriers or diluents. The compounds of the 
present invention can also be made into pharmaceutical preparations by any 
of the conventional methods giving solid, semisolid, liquid or gaseous 
forms for oral or parenteral administration. 
In manufacturing such preparations, the compounds of the present invention 
may be used in the form of their pharmaceutically acceptable salts. The 
compounds of the present invention may be used either solely or jointly in 
the form of a suitable combination. Alternatively, the compounds may be 
compounded with other pharmaceutically active components. 
In the case of oral preparations, the compounds of the present invention 
may be used alone or combined with appropriate additives to make tablets, 
diluted powders, granules or capsules. The compounds may be combined with 
conventional fillers such as lactose, mannitol, corn starch, and potato 
starch; binders such as crystalline cellulose, cellulose derivatives, gum 
arabic, corn starch and gelatin; lubricants such as talc and magnesium 
stearate; disintegrators such as corn starch, potato starch or sodium 
carboxymethylcellulose; and if desired with diluents, buffering agents, 
extenders, moisturizers, preservatives, flavoring agents and perfumes. 
Alternatively, the compounds of the present invention may be made into a 
suppository by mixing with a variety of bases. Exemplary bases include 
fatty and oil bases such as cocoa butter, emulsifying bases, water-soluble 
bases such as Macrogol and hydrophilic bases. 
In the case of injections, the compounds may be dissolved, suspended or 
emulsified in aqueous solvents or nonaqueous solvents. Examples of aqueous 
and nonaqueous solvents include distilled water, physiological saline 
liquid, Ringer solution and solutions containing plant oil, synthetic 
fatty acid glycerides, vegetable oil, higher fatty acid esters and 
propylene glycol. 
Further, depending upon the state of the patient, or the type of the 
disease, the compounds may be made into other preparation forms which are 
most suitable for the therapy such as inhalants, aerosols, ointments, 
poultices and eye drops. In the case of inhalations or aerosol 
preparations, the compounds of the invention in the form of a liquid or a 
minute powder can be filled up in an aerosol container with gas or a 
liquid spraying agent, and if desired, with conventional adjuvants such as 
humidifying agents or dispersing agents. 
Cataplasms can be prepared by mixing the compounds with mentha oil, 
concentrated glycerin, kaolin or other such additives. 
The desired doses of the compounds of the present invention vary depending 
upon the patient to be treated, the preparation form, the method of 
administration, and the period of administration. In general, 0.1 to 5,000 
mg or, preferably, 0.2 to 3,000 mg per day may be given to an adult by 
oral route for achieving the desired effect. 
In the case of parenteral administrations such as injections, doses of the 
compounds on the order of one third to one tenth of the above dose are 
preferable as daily doses.