U.S. patent application number 10/557541 was filed with the patent office on 2007-02-22 for immunosuppresive effects of pteridine derivatives.
Invention is credited to Piet Andre Maurits Maria Herdewijn, Wolfgang Eugen Pfleiderer, Mark Jozef Albert Waer.
Application Number | 20070043000 10/557541 |
Document ID | / |
Family ID | 33098022 |
Filed Date | 2007-02-22 |
United States Patent
Application |
20070043000 |
Kind Code |
A1 |
Waer; Mark Jozef Albert ; et
al. |
February 22, 2007 |
Immunosuppresive effects of pteridine derivatives
Abstract
Novel poly-substituted pteridinediones (lumazines), and mono- or
polysubstituted 2-thiolumazines, 4-thiolumazines or
2,4-dithiolumazines, having disclosed substituents in positions 1,
3, 6 and 7 of the pteridine ring, and pharmaceutically acceptable
salts thereof, are useful as biologically active ingredients in
preparing pharmaceutical compositions especially for the treatment
or prevention of a CNS disorder, a cell proliferative disorder, a
viral infection, an immune or auto-immune disorder or a transplant
rejection. Combinations of the pteridine derivatives of the
invention with an immunosuppressant or immunomodulator drug, an
antineoplastic drug or an antiviral agent, providing potential
synergistic effects, are also disclosed.
Inventors: |
Waer; Mark Jozef Albert;
(Heverlee, BE) ; Herdewijn; Piet Andre Maurits Maria;
(Rotselaar/Wezemaal, BE) ; Pfleiderer; Wolfgang
Eugen; (Konstanz, DE) |
Correspondence
Address: |
CLARK & ELBING LLP
101 FEDERAL STREET
BOSTON
MA
02110
US
|
Family ID: |
33098022 |
Appl. No.: |
10/557541 |
Filed: |
May 21, 2004 |
PCT Filed: |
May 21, 2004 |
PCT NO: |
PCT/EP04/05501 |
371 Date: |
November 21, 2005 |
Current U.S.
Class: |
514/81 ;
514/234.5; 514/251; 544/114; 544/244; 544/257 |
Current CPC
Class: |
A61P 25/00 20180101;
A61K 31/505 20130101; A61K 31/52 20130101; A61K 31/52 20130101;
A61K 38/13 20130101; A61P 37/02 20180101; C07D 475/02 20130101;
A61K 31/57 20130101; A61K 31/535 20130101; A61K 31/57 20130101;
A61K 38/13 20130101; A61P 9/00 20180101; A61P 35/00 20180101; C07F
9/6561 20130101; A61K 31/505 20130101; A61K 2300/00 20130101; A61K
2300/00 20130101; A61P 37/06 20180101; A61K 2300/00 20130101; A61K
2300/00 20130101; A61K 2300/00 20130101; A61K 31/535 20130101 |
Class at
Publication: |
514/081 ;
514/251; 544/244; 544/257; 514/234.5; 544/114 |
International
Class: |
A61K 31/675 20060101
A61K031/675; A61K 31/5377 20070101 A61K031/5377; A61K 31/525
20070101 A61K031/525; C07D 475/04 20070101 C07D475/04; C07F 9/6512
20070101 C07F009/6512 |
Foreign Application Data
Date |
Code |
Application Number |
May 23, 2003 |
US |
10/444,158 |
Dec 24, 2003 |
EP |
03079183.4 |
Claims
1-17. (canceled)
18. A poly-substituted pteridinedione being represented by the
formula (I), ##STR6## wherein Y.sub.1 and Y.sub.2 are both oxygen,
and none of R.sub.3 and R.sub.4 is hydrogen, and: R.sub.2 is a
radical selected from the group consisting of C.sub.1-7 alkyl;
C.sub.2-7 alkenyl; aryl; alkylaryl; .omega.-hydroxy C.sub.1-7
alkyl; .omega.-epoxy C.sub.1-7 alkyl; .omega.-carboxy C.sub.1-7
alkyl (wherein the carboxy group may be acid, ester, thioester,
acid halide or amide); .omega.-cyano C.sub.1-7 alkyl; arylalkyl;
arylalkenyl; heterocyclic-substituted alkyl;
heterocyclic-substituted alkenyl; groups having the formula --S--R
(i.e. wherein a sulfur atom is attached to the nitrogen atom of the
pteridine ring) wherein R is a monovalent group selected from the
group consisting of C.sub.1-7 alkyl, aryl and C.sub.3-10 cycloalkyl
and wherein the said monovalent group is optionally substituted
with one or more substituents selected from the group consisting of
amino, amino-acid, alkylamino, arylamino, cycloalkylamino,
carboxylic acid, carboxylic ester, sulfonic acid and phosphonic
acid; and optionally substituted heterocyclic radicals; R.sub.1 is
a radical independently defined as R.sub.2, or is hydrogen; R.sub.3
and R.sub.4 are independently selected from halogen and aryl
substituted with one or more substituents selected from the group
consisting of halogen, C.sub.1-4 alkyl, C.sub.2-7 alkenyl,
C.sub.2-7 alkynyl, C.sub.1-4 haloalkyl, C.sub.1-4 alkoxy, hydroxyl,
sulfhydryl, amino, C.sub.3-10 cycloalkoxy, aryloxy, arylalkyloxy,
oxyheterocyclic, heterocyclic-substituted alkyloxy, thio C.sub.1-7
alkyl, thio C.sub.3-10 cycloalkyl, thioaryl, thiohetero-cyclic,
arylalkylthio, heterocyclic-substituted alkylthio, formyl,
hydroxylamino, cyano, carboxylic acid or esters or thioesters or
amides thereof, thiocarboxylic acid or esters or thioesters or
amides thereof, C.sub.1-7 alkylamino, cycloalkyl-amino,
alkenylamino, cycloalkenylamino, alkynylamino, arylamino,
arylalkyl-amino, hydroxyalkylamino, mercaptoalkylamino,
heterocyclic amino, hydra-zino, alkylhydrazino and phenylhydrazino;
or a pharmaceutically acceptable salt or an enantiomer thereof.
19. A poly-substituted pteridinedione according to claim 18,
wherein R.sub.1 and R.sub.2 are independently selected from the
group consisting of benzyl, phenyl, 2-phenylethyl, butyric acid,
butyric acid ester, butyronitrile, 2-hydroxyethyl,
2-morpholinoethyl, 2-piperidinoethyl, 2-pyrrolidinoethyl, ethyl
acetate, 4-butyramido, N-methyl-4-butyramido,
N-propyl-4-butyramido, ethyl and methyl.
20. A poly-substituted pteridinedione according to claim 18,
wherein R.sub.4 is chloro or bromo.
21. A poly-substituted pteridinedione according to claim 19,
wherein R.sub.4 is chloro or bromo.
22. A poly-substituted pteridinedione according to claim 18,
wherein R.sub.3 is selected from the group consisting of
4-fluorophenyl, 4-chlorophenyl, 3,4-dichlorophenyl,
2,6-diisopropyl-4-bromophenyl, pentafluorophenyl,
4-trifluoromethylphenyl, 4-cyanophenyl, 2,6-dichlorophenyl,
2-fluorophenyl, 3-methoxyphenyl, 3,5-dichlorophenyl,
3,4-dimethoxyphenyl, 4-methylphenyl, 2,6-dimethoxyphenyl,
2-chlorophenyl, 3-chlorophenyl and 4-hydroxyphenyl.
23. A poly-substituted pteridinedione according to claim 18,
wherein R.sub.1 is hydrogen and R.sub.2 is selected from the group
consisting of benzyl, methyl and ethyl.
24. A poly-substituted pteridinedione according to claim 18, being
selected from the group consisting of:
1-methyl-6-(4'-methoxyphenyl)-7-chloro-lumazine,
1-methyl-6-(4'-methylphenyl)-7-chloro-lumazine,
1-methyl-6-(3',4'-dimethoxyphenyl)-7-chloro-lumazine,
1-methyl-6-(4'-chlorophenyl)-7-chloro-lumazine,
7-chloro-6-(4-methoxyphenyl)-1-methyl-lumazine,
7-chloro-6-(3,4-dimethoxyphenyl)-1-methyl-lumazine,
7-chloro-6-(4-fluorophenyl)-1-methyl-lumazine,
7-chloro-6-(3-methoxyphenyl)-1-methyl-lumazine,
7-chloro-(2,6-dimethoxyphenyl)-1-methyl-lumazine,
7-chloro-(2-chlorophenyl)-1-methyl-lumazine,
7-chloro-3-(chlorophenyl)-1-methyl-lumazine,
7-chloro-6-(4-cyanophenyl)-1-methyl-lumazine,
7-bromo-6-(3,4-dimethoxyphenyl)-1-methyl-lumazine,
7-bromo-6-(4-methoxyphenyl)-1-methyl-lumazine,
7-chloro-1-methyl-6-(4-methylphenyl)-lumazine,
7-chloro-6-(4-chlorophenyl)-1-methyl-lumazine,
7-chloro-6-(3,4-dimethoxyphenyl)-3-(ethyl
butyrate)-1-methyl-lumazine,
7-chloro-6-(3,4-dimethoxyphenyl)-3-(2-hydroxyethyl)-1-methyllumazine,
7-chloro-3-(ethyl butyrate)-1-methyl-6-(4-methoxyphenyl)-lumazine,
7-chloro-1,3-dimethyl-6-(4-methoxyphenyl)-lumazine,
7-bromo-6-(3,4-dimethoxyphenyl)-3-(ethylbutyrate)-1-methyl-lumazine,
7-bromo-3-(ethylbutyrate)-1-methyl-6-(4-methoxyphenyl)-lumazine,
7-bromo-1-methyl-3-(2-morpholinoethyl)-6-(3,4dimethoxyphenyl)-lumazine,
3-(ethyl butyrate)-7-fluoro-1-methyl-6-(4-methoxyphenyl)-lumazine,
7-bromo-1,3-dimethyl-6-(4-hydroxyphenyl)-lumazine, and
7-bromo-6-(3,4-dimethoxyphenyl)-3-(isopropyl
butyrate)-1-methyl-lumazine.
25. A pharmaceutical composition comprising one or more
pharmaceutically acceptable carriers and a poly-substituted
pteridinedione being represented by the formula (I), ##STR7##
wherein Y.sub.1 and Y.sub.2 are both oxygen, and none of R.sub.3
and R.sub.4 is hydrogen, and: R.sub.2 is a radical selected from
the group consisting of C.sub.1-7 alkyl; C.sub.2-7 alkenyl; aryl;
alkylaryl; .omega.-hydroxy C.sub.1-7 alkyl; .omega.-epoxy C.sub.1-7
alkyl; .omega.-carboxy C.sub.1-7 alkyl (wherein the carboxy group
may be acid, ester, thioester, acid halide or amide); .omega.-cyano
C.sub.1-7 alkyl; arylalkyl; arylalkenyl; heterocyclic-substituted
alkyl; heterocyclic-substituted alkenyl; groups having the formula
--S--R (i.e. wherein a sulfur atom is attached to the nitrogen atom
of the pteridine ring) wherein R is a monovalent group selected
from the group consisting of C.sub.1-7 alkyl, aryl and C.sub.3-10
cycloalkyl and wherein the said monovalent group is optionally
substituted with one or more substituents selected from the group
consisting of amino, amino-acid, alkylamino, arylamino,
cycloalkylamino, carboxylic acid, carboxylic ester, sulfonic acid
and phosphonic acid; and optionally substituted heterocyclic
radicals; R.sub.1 is a radical independently defined as R.sub.2, or
is hydrogen; R.sub.3 and R.sub.4 are independently selected from
halogen and aryl substituted with one or more substituents selected
from the group consisting of halogen, C.sub.1-4 alkyl, C.sub.2-7
alkenyl, C.sub.2-7 alkynyl, C.sub.1-4 haloalkyl, C.sub.1-4 alkoxy,
hydroxyl, sulfhydryl, amino, C.sub.3-10 cycloalkoxy, aryloxy,
arylalkyloxy, oxyheterocyclic, heterocyclic-substituted alkyloxy,
thio C.sub.1-7 alkyl, thio C.sub.3-10 cycloalkyl, thioaryl,
thiohetero-cyclic, arylalkylthio, heterocyclic-substituted
alkylthio, formyl, hydroxylamino, cyano, carboxylic acid or esters
or thioesters or amides thereof, thiocarboxylic acid or esters or
thioesters or amides thereof, C.sub.1-7 alkylamino,
cycloalkyl-amino, alkenylamino, cycloalkenylamino, alkynylamino,
arylamino, arylalkyl-amino, hydroxyalkylamino, mercaptoalkylamino,
heterocyclic amino, hydra-zino, alkylhydrazino and phenylhydrazino;
or a pharmaceutically acceptable salt or an enantiomer thereof.
26. A pharmaceutical composition according to claim 25, further
comprising one or more biologically-active drugs selected from the
group consisting of immunosuppressant and/or immunomodulator drugs,
antineoplastic drugs, and antiviral agents.
27. A pharmaceutical composition according to claim 25, further
comprising an immunomodulator drug selected from the group
consisting of acemannan, amiprilose, bucillamine, ditiocarb sodium,
imiquimod, Inosine Pranobex, interferon-.beta., interferon-.gamma.,
lentinan, levamisole, pidotimod, romurtide, platonin, procodazole,
propagermanium, thymomodulin, thymopentin and ubenimex.
28. A pharmaceutical composition according to claim 25, further
comprising an immunosuppressant drug selected from the group
consisting of cyclosporin A, substituted xanthines, pentoxyfylline,
tacrolimus, rapamycin, leflunomide, malononitrilamides,
mycophenolic acid and salts thereof, adrenocortical steroids,
azathioprine, brequinar, gusperimus, 6-mercaptopurine, mizoribine,
chloroquine, hydroxychloroquine and monoclonal antibodies with
immunosuppressive properties.
29. A pharmaceutical composition according to claim 25, further
comprising an antineoplastic drug selected from the group
consisting of alkaloids, alkylating agents, alkyl sulfonates,
aziridines, ethylenimines, methylmelamines, nitrogen mustards,
nitrosoureas, antibiotics, antimetabolites, folic acid analogues,
purine analogues and pyrimidine analogues, enzymes, interferon and
platinum complexes.
30. A pharmaceutical composition according to claim 25, further
comprising an antiviral agent selected from the group consisting of
HIV-1 IN inhibitors, nucleoside reverse transcriptase inhibitors,
zidovudine, lamivudine, didanosine, stavudine, zalcitabine,
non-nucleoside reverse transcriptase inhibitors, nevirapine,
delavirdine, foscarnet sodium, HIV-1 protease inhibitors,
saquinavir, ritonavir, indinavir, nelfinavir, acyclovir, cidofovir,
cytarabine, edoxudine, famciclovir, floxuridine, ganciclovir,
idoxuridine, penciclovir, sorivudine, trifluridine, valaciclovir,
vidarabine, kethoxal, methisazone, moroxydine, podophyllotoxin,
ribavirine, rimantadine, stallimycine, statolon, tromantadine and
xenazoic acid.
31. A method of prevention or treatment of a pathologic condition
selected from the group consisting of: transplant rejections and
autoimmune disorders, cardiovascular disorders, disorders of the
central nervous system, and cell proliferative disorders, said
method comprising administering a therapeutically effective amount
of a poly-substituted pteridinedione being represented by the
formula (I) ##STR8## wherein Y.sub.1 and Y.sub.2 are both oxygen,
and none of R.sub.3 and R.sub.4 is hydrogen, and: R.sub.2 is a
radical selected from the group consisting of C.sub.1-7 alkyl;
C.sub.2-7 alkenyl; aryl; alkylaryl; .omega.-hydroxy C.sub.1-7
alkyl; .omega.-epoxy C.sub.1-7 alkyl; .omega.-carboxy C.sub.1-7
alkyl (wherein the carboxy group may be acid, ester, thioester,
acid halide or amide); .omega.-cyano C.sub.1-7 alkyl; arylalkyl;
arylalkenyl; heterocyclic-substituted alkyl;
heterocyclic-substituted alkenyl; groups having the formula --S--R
(i.e. wherein a sulfur atom is attached to the nitrogen atom of the
pteridine ring) wherein R is a monovalent group selected from the
group consisting of C.sub.1-7 alkyl, aryl and C.sub.3-10 cycloalkyl
and wherein the said monovalent group is optionally substituted
with one or more substituents selected from the group consisting of
amino, amino-acid, alkylamino, arylamino, cycloalkylamino,
carboxylic acid, carboxylic ester, sulfonic acid and phosphonic
acid; and optionally substituted heterocyclic radicals; R.sub.1 is
a radical independently defined as R.sub.2, or is hydrogen; R.sub.3
and R.sub.4 are independently selected from halogen and aryl
substituted with one or more substituents selected from the group
consisting of halogen, C.sub.1-4 alkyl, C.sub.2-7 alkenyl,
C.sub.2-7 alkynyl, C.sub.1-4 haloalkyl, C.sub.1-4 alkoxy, hydroxyl,
sulfhydryl, amino, C.sub.3-10 cycloalkoxy, aryloxy, arylalkyloxy,
oxyheterocyclic, heterocyclic-substituted alkyloxy, thio C.sub.1-7
alkyl, thio C.sub.3-10 cycloalkyl, thioaryl, thiohetero-cyclic,
arylalkylthio, heterocyclic-substituted alkylthio, formyl,
hydroxylamino, cyano, carboxylic acid or esters or thioesters or
amides thereof, thiocarboxylic acid or esters or thioesters or
amides thereof, C.sub.1-7 alkylamino, cycloalkyl-amino,
alkenylamino, cycloalkenylamino, alkynylamino, arylamino,
arylalkyl-amino, hydroxyalkylamino, mercaptoalkylamino,
heterocyclic amino, hydra-zino, alkylhydrazino and phenylhydrazino;
or a pharmaceutically acceptable salt or an enantiomer thereof.
32. The method of prevention or treatment according to claim 31,
wherein said poly-substituted pteridinedione is administered in
combination with one or more biologically-active drugs selected
from the group consisting of immunosuppressant and/or
immunomodulator drugs, antineoplastic drugs, and antiviral agents.
Description
[0001] The invention relates to a class of novel poly-substituted
pteridine-2,4-diones (lumazines), as well as novel mono- and
polysubstituted 2-thiolumazines, 4-thiolumazines and
2,4-dithiolumazines. The invention further relates to
pharmaceutical compositions including a broad class of
poly-substituted pteridine-2,4-diones (lumazines), as well as mono-
and polysubstituted 2-thiolumazines, 4-thiolumazines and
2,4-dithiolumazines especially for the prevention and/or the
treatment of pathologic conditions such as, but not limited to,
immune and autoimmune disorders, organ and cells transplant
rejections, cell proliferative disorders, cardiovascular disorders,
disorders of the central nervous system and viral diseases.
[0002] The invention further relates to combined pharmaceutical
preparations comprising one or more polysubstituted
pteridine-2,4-diones (lumazines), as well as mono- and
polysubstituted 2-thiolumazines, 4-thiolumazines and
2,4-dithiolumazines and one or more known immunosuppressant drugs
or antineoplastic drugs or anti-viral drugs.
[0003] This invention also relates to a method for the prevention
and/or treatment of pathologic conditions such as, but not limited
to, immune and autoimmune disorders, organ and cells transplant
rejections, cell proliferative disorders, cardiovascular disorders,
disorders of the central nervous system and viral diseases by the
administration of an effective amount of a polysubstituted
pteridine-2,4-dione (lumazine), or a mono- or polysubstituted
2-thiolumazine, 4-thiolumazine or 2,4-dithio-lumazine optionally
combined with one or more known immunosuppressant drugs or
antineoplastic drugs or anti-viral drugs. Finally the invention
relates to a method for selecting or classifying biologically
active polysubstituted pteridine-2,4-diones (lumazines), as well as
mono- and polysubstituted 2-thiolumazines, 4-thiolumazines and
2,4dithiolumazines based on the determination of two or more in
vitro tests such as TNF-.alpha. and IL-1 .beta. assays.
BACKGROUND OF THE INVENTION
[0004] 2,4-dioxo-1,2,3,4-tetrahydropteridine is well known in the
art under the name lumazine. Gabriel and Sonn first disclosed in
Ber. Deut. Chem. Ges. (1907) 40:4850 making lumazine from
pyrazin-bicarboxamide. Timmis in Nature (1949) 164:139 disclosed
the synthesis of 1,3-dimethyl-6-phenyllumazine and
1,3-dimethyl-7-phenyl-lumazine by condensing a
6-amino-5-nitroso-pyrimidine with benzaldehyde or
methylphenylketone respectively. Zondler et al. in J. Heterocyclic
Chem. (1967) 4:124 and Taylor et al. in Heterocycles (1978) 10:37
disclosed 1,3,6-trimethyllumazine and 1,3-dimethyl-6-ethyllumazine.
Yoneda and Higuchi in J. Chem. Soc. Perkin (1977) 1336 disclosed
the preparation of various 1,3-dimethyl-6-aryllumazines starting
from 6-amino-1,3-dimethyl-5-aryliden-aminouracil. Kang et al. in J.
Heterocycl. Chem. (1987) 24:597-601 disclosed reacting
5,6-diamino-1,3-dimethyluracil either with
propanetrione-1,3-dioxime followed by cyclization to form
1,3-dimethyllumazine-6-carboxaldoxime, or with oximinoacetone
followed by cyclization to form 1,3,6-trimethyllumazine, or else
with methylglyoxal to form 1,3,7-trimethyllumazine. Both latter
compounds may easily, through acid hydrolysis in the presence of
formaldehyde, be converted into the corresponding
1,3-dimethyllumazine-carboxaldehydes which, due to their high
carbonyl reactivity, may in turn be converted into other lumazine
derivatives. Blicke et al. in J.A.C.S (1954) 76:2798-2800 disclosed
1,3-dimethyl-7-aminolumazine, 1,3,6,7-tetramethyllumazine,
1,3-dimethyl6,7-dihydroxy-lumazine and
1,3-dimethyl-6,7-diphenylumazine; Pfleiderer in Chem. Ber. (1957)
90:2588 disclosed 1,3-dimethyl-6-hydroxylumazine and
1,3-dimethyl-7-hydroxylumazine; Pfleiderer et al. in Chem. Ber.
(1973) 106:3149-3174 disclosed
1,3-dimethyl6-hydroxy-7-phenyllumazine,
1,3-dimethyl-6-phenyl-7-hydroxy-lumazine and
1,3-dimethyl-6,7-diisopropylumazine; Hutzenlaub et al. in Chem.
Ber. (1973) 106:3203-3215 disclosed 1,3-dimethyl-7-methoxylumazine,
1,3,6-trimethyl-7-hydroxy-lumazine and
1,3,6-trimethyl-7-methoxylumazine; Steppan et al. in Liebigs Ann.
Chem. (1982) 2135-2145 disclosed 1,3dimethyl-6-aminolumazine,
1,3-dimethyl-6-chlorolumazine, 1,3-dimethyl-7-chlorolumazine and
1,3-dimethyl-7-methylaminolumazine; Kasimierczuk et al. in Chem.
Ber. (1979) 112:1499-1513 disclosed 1,3-dimethyl-7-mercaptolumazine
and 1,3-dimethyl-7-methylthio-lumazine as well as a few substituted
2- or 4-thiolumazines and 2,4-dithiolumazines, starting from
substituted 6-amino-2-thiouracil or 6-amino-2,4-dithiouracil;
Eisele et al. in Pteridines (1993) 4:178-186 disclosed
1,3,6-trimethyllumazine-7-carboxylic acid and its methyl and ethyl
esters. Perez-Rubalcaba et al. in Liebigs Ann. Chem. (1983) 852-859
disclosed substituted 3-methyllumazines wherein one of the 6- and
7-substituents is phenyl whereas the other is chloro. Finally,
Weisenfeldt (1987) disclosed a series of tetra-substituted
lumazines wherein the 1- and 3-substituents are methyl and one of
the 6- and 7-substituents is chloro. Further, Fink et al. in Chem.
Berichte (1963) 96:2950-2963, as well as Pfleiderer,
Perez-Rubalcaba and Eisele (all cited supra) disclosed bi- and
tri-substituted lumazines wherein only one of the 1- and 3-nitrogen
atoms is substituted. Interestingly, none of the above-cited
substituted lumazines, 2-thiolumazines and 2,4-dithiolumazines was
ever said to have any kind of biological activity.
[0005] A few other substituted pteridine-2,4-diones (lumazines) are
already known in the art as being useful in the preparation of
medicines. For instance, U.S. Pat. No. 3,071,587 teaches
cyanoethylpteridinediones having central nervous system
(hereinafter referred as CNS) activity and anti-depressant
properties. WO 94/06431 teaches a
1-methyl-3-(10,11-epoxyundecyl)pteridine-dione being able to
inhibit IL-1 receptors, decrease proliferation of tumor & other
cells, stimulate hematopoeisis, suppress T-cell activation,
secretion of antibodies by B-cells and activation of macrophage or
endothelial cells by endotoxins, tumor necrosis factor (hereinafter
TNF), IL-1 or GM-CSF and enhance resistance of mesenchymal cells to
TNF. WO 94/11001 teaches 1-methyl-3-(hydroxy- and
dihydroxy-C.sub.9-25 alkyl) pteridinediones being able to inhibit
lysophosphatidic acid transferase as well as immune or cellular
response to stimuli, and therefore can be used to treat tumor
progression or invasion, autoimmune diseases, acute allergic
reactions mediated by TNF or IL-1, rheumatoid arthritis,
osteoarthritis, multiple sclerosis, diabetes, atherosclerosis,
restenosis, stroke, HIV infection, inflammatory response, septic
shock, CNS and bone diseases. Cottam et al. in J. Med. Chem. (1996)
39:2-9 and WO 98/52948 both disclose a
1-methyl-3-n-hexyl-6-carboxymethyl-7-carboxymethyl pteridine-dione
which, although included in a biological evaluation study of
inhibitors of TNF-.alpha., was not tested for TNF-.alpha. activity.
WO 96/20710 teaches substituted pteridinediones which inhibit
cellular responses to ceramide metabolites of the sphingomyelin
signal transduction pathway, inhibit inflammatory response
associated with TNF-.alpha. and fibroblast proliferation or
UV-induced cutaneous immune suppression and therefore can be used
to treat cirrhosis, cell senescence and apoptosis.
[0006] WO 00/45800 discloses the immunosuppressive effects of
pharmaceutical compositions for the treatment of autoimmuno
disorders and/or for the treatment or prevention of transplant
rejections comprising a pteridine derivative of general formula:
##STR1## wherein:
[0007] R.sub.1 and R.sub.2 are independently hydrogen; aliphatic
saturated or unsaturated, straight or branched carbon chains with 1
to 7 carbon atoms; substituted or unsubstituted aryl or alkylaryl
substituents, whereby the carbon atoms may be oxidized represented
by alcohol or carbonyl function or carboxylic acids and their
esters;
[0008] R.sub.3 and R.sub.4 are independently hydrogen, hydroxyl,
halogen, alkyl, haloalkyl, alkoxy, wherein the alkyl group may be
branched or straight and contains one or four carbon atoms, formyl
and derivatives such as hydroxylamino conjugates and acetals,
cyano, carboxylic acids and carboxyl acid derivatives such as
esters and amides, sulfhydryl, amino, alkylamino, cycloalkylamino,
alkenylamino, alkynyl-amino, benzylamino, hydroxylalkylamino,
morpholinoalkylamino, phenylhydra-zino, morpholino, piperidino,
mercaptobenzyl, mercaptoalkyl, cysteinyl ester, styryl, aromatic
ring; aromatic or heterocyclic substituent substituted with an
aliphatic spacer between the pteridine ring and the aromatic
substituent of 1 to 4 carbon atoms, whereby said spacer may contain
an alcohol function, carbonyl function, halogen, ether, and may be
saturated or unsaturated; branched or straight, saturated or
unsaturated aliphatic chain of 1 to 7 carbon atoms which may
contain one or more functions chosen from the group comprising
carbonyl, alcohol, ether, carboxyester nitro, thioalkyl, halogen;
and
[0009] Y.sub.1 and Y.sub.2 are both oxygen,
or a pharmaceutical salt thereof, and a pharmaceutically acceptable
carrier.
[0010] WO 00/45800 also discloses a compound,
1,3-dimethyl-6-benzoyl-7-(4-methoxyphenyl)lumazine, which has poor
results in a Mixed Lymphocyte Reaction test as well as in CD3 and
CD 28 assays.
[0011] WO 03/067257 discloses making 3-methyl6-iminouracil in a
three-steps procedure starting from O-methylisourea hydrochloride
and methylcyanoacetate with a combined yield of 29%, followed by
conversion into 5,6-diamino-3-methyluracil. C. Muller et al. in J.
Med. Chem. (2002) 45:3440-3450 discloses 5,6-diamino-1-benzyluracil
and a procedure for making it.
[0012] Nevertheless, there still is a need in the art for specific
and highly therapeutically active compounds, such as, but not
limited to, drugs for treating immune and autoimmune disorders,
organ and cells transplant rejections, cell proliferative
disorders, cardiovascular disorders, disorders of the central
nervous system and viral diseases. In particular, there is a need
in the art to provide immunosuppressive compounds or antineoplastic
drugs or anti-viral drugs which are active in a minor dose in order
to replace existing drugs having significant side effects and to
decrease treatment costs.
[0013] Currently used immunosuppressive drugs include
antiproliferative agents, such as methotrexate, azathioprine, and
cyclophosphamide. Since these drugs affect mitosis and cell
division, they have severe toxic effects on normal cells with high
turn-over rate such as bone marrow cells and the gastrointestinal
tract lining. Accordingly, marrow depression and liver damage are
common side effects.
[0014] Anti-inflammatory compounds used to induce immunosuppression
include adrenocortical steroids such as dexamethasone and
prednisolone. The common side effects observed with the use of
these compounds are frequent infections, abnormal metabolism,
hypertension, and diabetes.
[0015] Other immunosuppressive compounds currently used to inhibit
lymphocyte activation and subsequent proliferation include
cyclosporine, tacrolimus and rapamycin. Cyclosporine and its
relatives are among the most commonly used immunosuppressant drugs.
Cyclosporine is typically used for preventing or treating organ
rejection in kidney, liver, heart, pancreas, bone marrow, and
heart-lung transplants, as well as for the treatment of autoimmune
and inflammatory diseases such as Crohn's disease, aplastic anemia,
multiple sclerosis, myasthenia gravis, uveitis, biliary cirrhosis,
etc. However, cyclosporines suffer from a small therapeutic dose
window and severe toxic effects including nephrotoxicity,
hepatotoxicity, hypertension, hirsutism, cancer, and neurotoxicity.
Another such drug, mitoxantrone, is known to induce heart and
kidney toxicity.
[0016] Additionally, monoclonal antibodies with immunosuppressant
properties, such as OKT3, have been used to prevent and/or treat
graft rejection. Introduction of such monoclonal antibodies into a
patient, as with many biological materials, induces several
side-effects, such as dyspnea. Within the context of many
life-threatening diseases, organ transplantation is considered a
standard treatment and, in many cases, the only alternative to
death. The immune response to foreign cell surface antigens on the
graft, encoded by the major histo-compatibility complex
(hereinafter referred as MHC) and present on all cells, generally
precludes successful transplantation of tissues and organs unless
the transplant tissues come from a compatible donor and the normal
immune response is suppressed. Other than identical twins, the best
compatibility and thus, long term rates of engraftment, are
achieved using MHC identical sibling donors or MHC identical
unrelated cadaver donors. However, such ideal matches are difficult
to achieve. Further, with the increasing need of donor organs an
increasing shortage of transplanted organs currently exists.
Accordingly, xenotransplantation has emerged as an area of
intensive study, but faces many hurdles with regard to rejection
within the recipient organism.
[0017] The host response to an organ allograft involves a complex
series of cellular interactions among T and B lymphocytes as well
as macrophages or dendritic cells that recognize and are activated
by foreign antigen. Co-stimulatory factors, primarily cytokines,
and specific cell-cell interactions, provided by activated
accessory cells such as macrophages or dendritic cells are
essential for T-cell proliferation. These macrophages and dendritic
cells either directly adhere to T-cells through specific adhesion
proteins or secrete cytokines that stimulate T-cells, such as IL-12
and IL-15. Accessory cell-derived co-stimulatory signals stimulate
activation of interleukin-2 (IL-2) gene transcription and
expression of high affinity IL-2 receptors in T-cells. IL-2 is
secreted by T lymphocytes upon antigen stimulation and is required
for normal immune responsiveness. IL-2 stimulates lymphoid cells to
proliferate and differentiate by binding to IL-2 specific cell
surface receptors (IL-2R). IL-2 also initiates helper T-cell
activation of cytotoxic T-cells and stimulates secretion of
interferon-.gamma. which in turn activates cytodestructive
properties of macrophages. Furthermore, IFN-.gamma. and IL-4 are
also important activators of MHC class II expression in the
transplanted organ, thereby further expanding the rejection cascade
by enhancing the immunogenicity of the grafted organ The current
model of a T-cell mediated response suggests that T-cells are
primed in the T-cell zone of secondary lymphoid organs, primarily
by dendritic cells. The initial interaction requires cell to cell
contact between antigen-loaded MHC molecules on antigen-presenting
cells (hereinafter referred as APC) and the T-cell receptor/CD3
complex on T-cells. Engagement of the TCR/CD3 complex induces CD154
expression predominantly on CD4 T-cells that in turn activate the
APC through CD40 engagement, leading to improved antigen
presentation. This is caused partly by upregulation of CD80 and
CD86 expression on the APC, both of which are ligands for the
important CD28 co-stimulatory molecule on T-cells. However,
engagement of CD40 also leads to prolonged surface expression of
MHC-antigen complexes, expression of ligands for 4-1BB and OX40
(potent co-stimulatory molecules expressed on activated T-cells).
Furthermore, CD40 engagement leads to secretion of various
cytokines (e.g., IL-12, IL-15, TNF-.alpha., IL-1, IL-6, and IL-8)
and chemokines, all of which have important effects on both APC and
T-cell activation and maturation. Similar mechanisms are involved
in the development of auto-immune disease, such as type I diabetes.
In humans and non-obese diabetic mice, insulin-dependent diabetes
mellitus results from a spontaneous T-cell dependent auto-immune
destruction of insulin-producing pancreatic .beta. cells that
intensifies with age. The process is preceded by infiltration of
the islets with mononuclear cells (insulitis), primarily composed
of T lymphocytes. A delicate balance between auto-aggressive
T-cells and suppressor-type immune phenomena determine whether
expression of auto-immunity is limited to insulitis or not.
Therapeutic strategies that target T-cells have been successful in
preventing further progress of the auto-immune disease. These
include neonatal thymectomy, administration of cyclosporine, and
infusion of anti-pan T-cell, anti-CD4, or anti-CD25 (IL-2R)
monoclonal antibodies. The aim of all rejection prevention and
auto-immunity reversal strategies is to suppress the patient's
immune reactivity to the antigenic tissue or agent, with a minimum
of morbidity and mortality. Accordingly, a number of drugs are
currently being used or investigated for their immunosuppressive
properties. As discussed above, the most commonly used
immunosuppressant is cyclosporine, which however has numerous side
effects. Accordingly, in view of the relatively few choices for
agents effective at immunosuppression with low toxicity profiles
and manageable side effects, there exists a need in the art for
identification of alternative immunosuppressive agents and for
agents acting as complement to calcineurin inhibition.
[0018] There is also a need in the art to improve therapeutic
efficiency by providing pharmaceutical compositions or combined
preparations exhibiting a synergistic effect as a result of
combining two or more immunosuppressant drugs, or antineoplastic
drugs or anti-viral drugs.
SUMMARY OF THE INVENTION
[0019] In a first embodiment, the present invention relates to a
group of novel poly-substituted pteridine-2,4-diones (lumazines),
as well as novel mono- and poly-substituted 2-thiolumazines,
4-thiolumazines and 2,4-dithiolumazines and their pharmaceutically
acceptable salts and enantiomers. These compounds may be
represented by the general formula (I): ##STR2## wherein: [0020] a)
if Y.sub.1 and Y.sub.2 are both oxygen and R.sub.4 is hydrogen,
then: [0021] R.sub.1 is a radical selected from the group
consisting of hydrogen; C.sub.1-5 alkyl; C.sub.2-7 alkenyl; aryl;
alkylaryl; .omega.-hydroxy C.sub.1-7 alkyl; .omega.-epoxy C.sub.1-7
alkyl; .omega.-carboxy C.sub.1-7 alkyl (wherein the carboxy group
may be acid, ester, thioester, acid halide or amide); .omega.-cyano
C.sub.1-7 alkyl; arylalkyl; arylalkenyl; heterocyclic-substituted
alkyl; heterocyclic-substituted alkenyl; groups having the formula
--S--R (i.e. wherein a sulfur atom is attached to the nitrogen atom
of the pteridine ring) wherein R is a monovalent group selected
from the group consisting of C.sub.1-7 alkyl, aryl and C.sub.3-10
cycloalkyl and wherein the said monovalent group is optionally
substituted with one or more substituents selected from the group
consisting of amino, amino-acid, alkylamino, arylamino,
cycloalkylamino, carboxylic acid, carboxylic ester, sulfonic acid
and phosphonic acid; and optionally substituted heterocyclic
radicals preferably selected from the group consisting of
diazepinyl, oxadiazinyl, thiadiazinyl, dithiazinyl, triazolonyl,
diazepinonyl, triazepinyl, triazepinonyl, tetrazepinonyl,
benzoquinolinyl, benzothiazinyl, benzothiazinonyl, benzoxathiinyl,
benzodioxinyl, benzodithiinyl, benzoxazepinyl, benzothiazepinyl,
benzodiazepinyl, benzodioxepinyl, benzodithiepinyl, benzoxazocinyl,
benzothiazocinyl, benzodiazocinyl, benzoxathiocinyl,
benzodioxocinyl, benzotrioxepinyl, benzoxathiazepinyl,
benzoxadiazepinyl, benzothiadiazepinyl, benzotriazepinyl,
benzoxathiepinyl, benzotriazinonyl, benzoxazolinonyl, azetidinonyl,
azaspiroundecyl, dithiaspirodecyl, selenazinyl, selenazolyl,
selenophenyl, hypoxanthinyl, azahypoxanthinyl, bipyrazinyl,
bipyridinyl, oxazolidinyl, diselenopyrimidinyl, benzopyrenyl,
benzopyranonyl, benzophenazinyl, benzoquinolizinyl,
dibenzocarbazolyl, dibenzoacridinyl, dibenzophenazinyl,
dibenzothiepinyl, dibenzooxepinyl, dibenzopyranonyl,
dibenzoquinoxalinyl, dibenzothiazepinyl, dibenzoisoquinolinyl,
tetraazaadamantyl, thiatetraazaadamantyl, oxauracil, oxazinyl,
dibenzothiophenyl, dibenzofuranyl, oxazolinyl, oxazolonyl,
azaindolyl, azolonyl, thiazolinyl, thiazolonyl, thiazolidinyl,
thiazanyl, pyrimidonyl, thiopyrimidonyl, thiamorpholinyl,
azlactonyl, naphtindazolyl, naphtindolyl, naphtothiazolyl,
naphtothioxolyl, naphtoxindolyl, naphtotriazolyl, naphtopyranyl,
indolinyl, indolizidinyl, oxabicycloheptyl, azabenzimidazolyl,
azacycloheptyl, azacyclooctyl, azacyclononyl, azabicyclononyl,
tetrahydropyranyl, tetrahydropyronyl, tetrahydroquinoleinyl,
tetrahydrothienyl and dioxide thereof, dihydrothienyl dioxide,
dioxindolyl, dioxinyl, dioxenyl, dioxazinyl, thioxanyl, thioxolyl,
thiourazolyl, thiotriazolyl, thiopyranyl, thiopyronyl, coumarinyl,
quinoleinyl, oxyquinoleinyl, quinuclidinyl, xanthinyl,
dihydropyranyl, benzodihydrofuryl, benzothiopyronyl,
benzothiopyranyl, benzoxazinyl, benzoxazolyl, benzodioxolyl,
benzodioxanyl, benzothiadiazolyl, benzotriazinyl, benzothiazolyl,
benzoxazolyl, phenothioxinyl, phenothiazolyl, phenothienyl,
phenopyronyl, phenoxazolyl, pyridinyl, dihydropyridinyl,
tetrahydropyridinyl, piperidinyl, morpholinyl, thiomorpholinyl,
pyrazinyl, pyrimidinyl, pyridazinyl, triazinyl, tetrazinyl,
triazolyl, benzotriazolyl, tetrazolyl, imidazolyl, pyrazolyl,
thiazolyl, thiadiazolyl, isothiazolyl, oxazolyl, oxadiazolyl,
pyrrolyl, furyl, dihydrofuryl, furoyl, hydantoinyl, dioxolanyl,
dioxolyl, dithianyl, dithienyl, dithiinyl, thienyl, indolyl,
indazolyl, benzofuryl, benzothienyl, quinolyl, quinazolinyl,
quinoxalinyl, carbazolyl, phenoxazinyl, phenothiazinyl, xanthenyl,
purinyl, benzothienyl, naphtothienyl, thianthrenyl, pyranyl,
pyronyl, benzo-pyronyl, isobenzofuranyl, chromenyl, phenoxathiinyl,
indolizinyl, quinolizinyl, isoquinolyl, phthalazinyl,
naphthiridinyl, cinnolinyl, pteridinyl, carbolinyl, acridinyl,
perimidinyl, phenanthrolinyl, phenazinyl, phenothiazinyl,
imidazo-linyl, imidazolidinyl, benzimidazolyl, pyrazolinyl,
pyrazolidinyl, pyrrolinyl, pyrrolidinyl, piperazinyl, uridinyl,
thymidinyl, cytidinyl, azirinyl, aziridinyl, diazirinyl,
diaziridinyl, oxiranyl, oxaziridinyl, dioxiranyl, thiiranyl,
azetyl, dihydroazetyl, azetidinyl, oxetyl, oxetanyl, thietyl,
thietanyl, oxetanonyl, diaza-bicyclooctyl, diazetyl,
diaziridinonyl, diaziridinethionyl, chromanyl, chroma-nonyl,
thiochromanyl, thiochromanonyl, thiochromenyl, benzofuranyl,
benzisothiazolyl, benzocarbazolyl, benzochromonyl,
benzisoalloxazinyl, benzocoumarinyl, thiocoumarinyl,
phenometoxazinyl, phenoparoxazinyl, phentriazinyl, thiodiazinyl,
thiodiazolyl, indoxyl, thioindoxyl, benzodiazinyl (e.g.
phtalazinyl), phtalidyl, phtalimidinyl, phtalazonyl, alloxazinyl,
dibenzo-pyronyl (i.e. xanthonyl), xanthionyl, isatyl, isopyrazolyl,
isopyrazolonyl, urazolyl, urazinyl, uretinyl, uretidinyl, succinyl,
succinimido, benzylsultimyl, and benzylsultamyl; [0022] R.sub.2 is
a radical selected from the group consisting of hydrogen; C.sub.1-5
alkyl; C.sub.2-7 alkenyl; aryl; alkylaryl; .omega.-hydroxy
C.sub.1-7 alkyl; .omega.-epoxy C.sub.1-7 alkyl; .omega.-carboxy
C.sub.1-7 alkyl (wherein the carboxy group may be acid, ester,
thioester, acid halide or amide); .omega.-cyano C.sub.1-7 alkyl;
arylalkyl; arylalkenyl; heterocyclic-substituted alkyl;
heterocyclic-substituted alkenyl; groups having the formula --S--R
(i.e. wherein a sulfur atom is attached to the nitrogen atom of the
pteridine ring) wherein R is a monovalent group selected from the
group consisting of C.sub.1-7 alkyl, aryl and C.sub.3-10 cycloalkyl
and wherein the said monovalent group is optionally substituted
with one or more substituents selected from the group consisting of
amino, amino-acid, alkylamino, arylamino, cycloalkylamino,
carboxylic acid, carboxylic ester, sulfonic acid and phosphonic
acid; and optionally substituted heterocyclic radicals preferably
other than tetrahydrofuryl, i.e. preferably selected from the group
consisting of diazepinyl, oxadiazinyl, thiadiazinyl, dithiazinyl,
triazolonyl, diazepinonyl, triazepinyl, triazepinonyl,
tetrazepinonyl, benzoquinolinyl, benzothiazinyl, benzothiazinonyl,
benzoxathiinyl, benzodioxinyl, benzodithiinyl, benzoxazepinyl,
benzothiazepinyl, benzodiazepinyl, benzodioxepinyl,
benzodithiepinyl, benzoxazocinyl, benzothiazocinyl,
benzodiazocinyl, benzoxathiocinyl, benzodioxocinyl,
benzotrioxepinyl, benzoxathiazepinyl, benzoxadiazepinyl,
benzothiadiazepinyl, benzotriazepinyl, benzoxathiepinyl,
benzotriazinonyl, benzoxazolinonyl, azetidinonyl, azaspiroundecyl,
dithiaspirodecyl, selenazinyl, selenazolyl, selenophenyl,
hypoxanthinyl, azahypoxanthinyl, bipyrazinyl, bipyridinyl,
oxazolidinyl, diselenopyrimidinyl, benzodioxocinyl, benzopyrenyl,
benzopyranonyl, benzophenazinyl, benzoquinolizinyl,
dibenzocarbazolyl, dibenzoacridinyl, dibenzophenazinyl,
dibenzothiepinyl, dibenzooxepinyl, dibenzopyranonyl,
dibenzoquinoxalinyl, dibenzothiazepinyl, dibenzoisoquinolinyl,
tetraazaadamantyl, thiatetraazaadamantyl, oxauracil, oxazinyl,
dibenzothiophenyl, dibenzofuranyl, oxazolinyl, oxazolonyl,
azaindolyl, azolonyl, thiazolinyl, thiazolonyl, thiazolidinyl,
thiazanyl, pyrimidonyl, thiopyrimidonyl, thiamorpholinyl,
aziactonyl, naphtindazolyl, naphtindolyl, naphtothiazolyl,
naphtothioxolyl, naphtoxindolyl, naphtotriazolyl, naphtopyranyl,
indolinyl, indolizidinyl, oxabicycloheptyl, azabenzimidazolyl,
azacycloheptyl, azacyclooctyl, azacyclononyl, azabicyclononyl,
tetrahydropyranyl, tetrahydropyronyl, tetrahydroquinoleinyl,
tetrahydrothienyl and dioxide thereof, dihydrothienyl dioxide,
dioxindolyl, dioxinyl, dioxenyl, dioxazinyl, thioxanyl, thioxolyl,
thio-urazolyl, thiotriazolyl, thiopyranyl, thiopyronyl, coumarinyl,
quinoleinyl, oxyquinoleinyl, quinuclidinyl, xanthinyl,
dihydropyranyl, benzodihydrofuryl, benzothiopyronyl,
benzothiopyranyl, benzoxazinyl, benzoxazolyl, benzodioxolyl,
benzodioxanyl, benzothiadiazolyl, benzotriazinyl, benzothiazolyl,
benzoxazolyl, phenothioxinyl, phenothiazolyl, phenothienyl,
phenopyronyl, phenoxazolyl, pyridinyl, dihydropyridinyl,
tetrahydropyridinyl, piperidinyl, morpholinyl, thiomorpholinyl,
pyrazinyl, pyrimidinyl, pyridazinyl, triazinyl, tetrazinyl,
triazolyl, benzotriazolyl, tetrazolyl, imidazolyl, pyrazolyl,
thiazolyl, thiadiazolyl, isothiazolyl, oxazolyl, oxadiazolyl,
pyrrolyl, furyl, dihydrofuryl, furoyl, hydantoinyl, dioxolanyl,
dioxolyl, dithianyl, dithienyl, dithiinyl, thienyl, indolyl,
indazolyl, benzofuryl, benzothienyl, quinolyl, quinazolinyl,
quinoxalinyl, carbazolyl, phenoxazinyl, phenothiazinyl, xanthenyl,
purinyl, benzothienyl, naphtothienyl, thianthrenyl, pyranyl,
pyronyl, benzopyronyl, isobenzofuranyl, chromenyl, phenoxathiinyl,
indolizinyl, quinolizinyl, isoquinolyl, phthalazinyl,
naphthiridinyl, cinnolinyl, pteridinyl, carbolinyl, acridinyl,
perimidinyl, phenanthrolinyl, phenazinyl, phenothiazinyl,
imidazolinyl, imidazolidinyl, benzimidazolyl, pyrazolinyl,
pyrazolidinyl, pyrrolinyl, pyrrolidinyl, piperazinyl, uridinyl,
thymidinyl, cytidinyl, azirinyl, aziridinyl, diazirinyl,
diaziridinyl, oxiranyl, oxaziridinyl, dioxiranyl, thiiranyl,
azetyl, dihydroazetyl, azetidinyl, oxetyl, oxetanyl, thietyl,
thietanyl, oxetanonyl, diaza-bicyclooctyl, diazetyl,
diaziridinonyl, diaziridinethionyl, chromanyl, chroma-nonyl,
thiochromanyl, thiochromanonyl, thiochromenyl, benzofuranyl,
benzisothiazolyl, benzocarbazolyl, benzochromonyl,
benzisoalloxazinyl, benzocoumarinyl, thiocoumarinyl,
phenometoxazinyl, phenoparoxazinyl, phentriazinyl, thiodiazinyl,
thiodiazolyl, indoxyl, thioindoxyl, benzodiazinyl (e.g.
phtalazinyl), phtalidyl, phtalimidinyl, phtalazonyl, alloxazinyl,
dibenzo-pyronyl (i.e. xanthonyl), xanthionyl, isatyl, isopyrazolyl,
isopyrazolonyl, urazolyl, urazinyl, uretinyl, uretidinyl, succinyl,
succinimido, benzylsultimyl, and benzylsultamyl; [0023] at most one
of R.sub.1 and R.sub.2 is hydrogen; and [0024] R.sub.3 is an atom
or radical selected from the group consisting of fluorine; iodine;
C.sub.3-4 alkyl; C.sub.2-7 alkenyl; C.sub.2-7 alkynyl; C.sub.3-4
haloalkyl; C.sub.1-2 haloalkyl wherein halo is fluoro or chloro;
C.sub.1-4 alkoxy; C.sub.3-10 cycloalkoxy; aryloxy; arylalkyloxy;
oxyheterocyclic; heterocyclic-substituted alkyloxy; thio C.sub.1-7
alkyl; thio C.sub.3-10 cycloalkyl; thioaryl; thioheterocyclic;
arylalkylthio; heterocyclic-substituted alkylthio; hydroxylamino;
acetal; carboxylic acid esters, thioesters and amides;
thiocarboxylic acid; thiocarboxylic acid esters, thioesters and
amides; sulfhydryl; C.sub.2-7 alkylamino; cycloalkylamino;
alkenylamino; cycloalkenylamino; alkynylamino; arylamino;
arylalkylamino; hydroxyalkylamino; mercaptoalkyl-amino;
heterocyclic amino; heterocyclic-substituted alkylamino; oximino;
alkyloximino; hydrazino; alkylhydrazino; phenylhydrazino; cysteinyl
acid, esters or amides; aryl substituted with one or more
substituents selected from the group consisting of halogen,
C.sub.1-4 alkyl, C.sub.3-7 alkenyl, C.sub.2-7 alkynyl, C.sub.1-4
haloalkyl, C.sub.2-4 alkoxy, hydroxyl, sulfhydryl, amino,
C.sub.3-10 cycloalkoxy, aryloxy, arylalkyloxy, oxyheterocyclic,
heterocyclic-substituted alkyloxy, thio C.sub.1-7 alkyl, thio
C.sub.3-10 cycloalkyl, thioaryl, thioheterocyclic, arylalkylthio,
heterocyclic-substituted alkylthio, formyl, hydroxylamino, cyano,
carboxylic acid or esters or thioesters or amides thereof,
thiocarboxylic acid or esters or thioesters or amides thereof,
C.sub.1-7 alkylamino, cycloalkylamino, alkenylamino,
cycloalkenylamino, alkynylamino, arylamino, arylalkylamino,
hydroxylalkyl-amino, mercaptoalkylamino, heterocyclic amino,
hydrazino, alkylhydrazino and phenylhydrazino; optionally
substituted heterocyclic radicals; aryl or heterocyclic radicals
substituted with an aliphatic spacer (linking group) between the
pteridine ring and the aryl or heterocyclic radical, whereby said
aliphatic spacer is a branched or straight, saturated or
unsaturated aliphatic chain of 2 to 4 carbon atoms which may
contain one or more functions, atoms or radicals selected from the
group consisting of carbonyl (oxo), alcohol (hydroxyl), ether,
acetal, amino, imino, oximino, alkyloximino, amino-acid, cyano,
carboxylic acid or ester or thioester or amide, nitro, thio
C.sub.1-7 alkyl, thio C.sub.3-10 cycloalkyl, C.sub.1-7 alkylamino,
cycloalkylamino, alkenylamino, cycloalkenylamino, alkynylamino,
arylamino, arylalkylamino, hydroxyalkyl-amino, mercaptoalkylamino,
heterocyclic amino, hydrazino, alkylhydrazino, phenylhydrazino,
sulfonyl, sulfonamido and halogen, or whereby said aliphatic spacer
is a methylene group containing a function, atom or radical chosen
from the group consisting of carbonyl (oxo), alcohol (hydroxyl),
ether, acetal, amino, imino, oximino, alkyloximino, amino-acid,
cyano, carboxylic acid or ester or thioester or amide, nitro, thio
C.sub.1-7 alkyl, thio C.sub.3-10 cycloalkyl, C.sub.1-7 alkylamino,
cycloalkylamino, alkenylamino, cycloalkenylamino, alkynylamino,
arylamino, arylalkylamino, hydroxyalkylamino, mercaptoalkyl-amino,
heterocyclic amino, hydrazino, alkylhydrazino, phenylhydrazino,
sulfonyl, sulfonamido and halogen; branched or straight, saturated
or unsaturated aliphatic chain of 3 to 7 carbon atoms containing
one or more functions selected from the group consisting of
carbonyl (oxo), alcohol (hydroxyl), ether, acetal, amino, imino,
oximino, alkyloximino, amino-acid, cyano, carboxylic acid ester or
amide, nitro, thio C.sub.1-7 alkyl, thio C.sub.3-10 cycloalkyl,
C.sub.1-7 alkylamino, cycloalkylamino, alkenylamino,
cycloalkenylamino, alkynylamino, arylamino, arylalkylamino,
hydroxylalkylamino, mercaptoalkyl-amino, heterocyclic amino,
hydrazino, alkylhydrazino, phenylhydrazino, sulfonyl, sulfonamido
and halogen; hydroxyethyl; oximinoethyl; alkyl-oximinoethyl; and
methyl or ethyl or ethenyl containing one or more atoms, functions
or radicals selected from the group consisting of ether, acetal,
amino, imino, amino-acid, cyano, carboxylic acid or ester or
thioester or amide, nitro, thio C.sub.1-7 alkyl, thio C.sub.3-10
cycloalkyl, C.sub.1-7 alkylamino, cyclo-alkylamino, alkenylamino,
cycloalkenylamino, alkynylamino, arylalkylamino, hydroxyalkylamino,
mercaptoalkylamino, heterocyclic amino, hydrazino, alkylhydrazino,
phenylhydrazino, sulfonyl, sulfonamido, fluoro and chloro; [0025]
b) if Y.sub.1 and Y.sub.2 are both oxygen and R.sub.3 is hydrogen,
then: [0026] R.sub.1 is a radical selected from the group
consisting of hydrogen; C.sub.1-5 alkyl; C.sub.2-7 alkenyl; aryl;
alkylaryl; .omega.-hydroxy C.sub.1-5 alkyl; .omega.-epoxy C.sub.1-5
alkyl; .omega.-carboxy C.sub.1-5 alkyl (wherein the carboxy group
may be acid, ester, thioester, acid halide or amide);
.omega.-cyano C.sub.1-7 alkyl; arylalkyl; arylalkenyl;
heterocyclic-substituted alkyl; heterocyclic-substituted alkenyl;
groups having the formula --S--R (i.e. wherein a sulfur atom is
attached to the nitrogen atom of the pteridine ring) wherein R is a
monovalent group selected from the group consisting of C.sub.1-7
alkyl, aryl and C.sub.3-10 cycloalkyl and wherein the said
monovalent group is optionally substituted with one or more
substituents selected from the group consisting of amino,
amino-acid, alkylamino, arylamino, cycloalkylamino, carboxylic
acid, carboxylic ester, sulfonic acid and phosphonic acid; and
optionally substituted heterocyclic radicals preferably selected
from the group consisting of diazepinyl, oxadiazinyl, thiadiazinyl,
dithiazinyl, triazolonyl, diazepinonyl, triazepinyl, triazepinonyl,
tetrazepinonyl, benzoquinolinyl, benzothiazinyl, benzothiazinonyl,
benzoxathiinyl, benzodioxinyl, benzodithiinyl, benzoxazepinyl,
benzothiazepinyl, benzodiazepinyl, benzodioxepinyl,
benzodithiepinyl, benzoxazocinyl, benzothiazocinyl,
benzodiazocinyl, benzoxathiocinyl, benzodioxocinyl,
benzotrioxepinyl, benzoxathiazepinyl, benzoxadiazepinyl,
benzothiadiazepinyl, benzo-triazepinyl, benzoxathiepinyl,
benzotriazinonyl, benzoxazolinonyl, azetidinonyl, azaspiroundecyl,
dithiaspirodecyl, selenazinyl, selenazolyl, selenophenyl,
hypoxanthinyl, azahypoxanthinyl, bipyrazinyl, bipyridinyl,
oxazolidinyl, diselenopyrimidinyl, benzopyrenyl, benzopyranonyl,
benzophenazinyl, benzoquinolizinyl, dibenzocarbazolyl,
dibenzoacridinyl, dibenzophenazinyl, dibenzothiepinyl,
dibenzooxepinyl, dibenzopyranonyl, dibenzoquinoxalinyl,
dibenzothiazepinyl, dibenzoisoquinolinyl, tetraaza-adamantyl,
thiatetraazaadamantyl, oxauracil, oxazinyl, dibenzothiophenyl,
dibenzofuranyl, oxazolinyl, oxazolonyl, azaindolyl, azolonyl,
thiazolinyl, thiazolonyl, thiazolidinyl, thiazanyl, pyrimidonyl,
thiopyrimidonyl, thiamorpholinyl, azlactonyl, naphtindazolyl,
naphtindolyl, naphtothiazolyl, naphtothioxolyl, naphtoxindolyl,
naphtotriazolyl, naphtopyranyl, indolinyl, indolizidinyl,
oxabicycloheptyl, azabenzimidazolyl, azacycloheptyl, azacyclooctyl,
azacyclononyl, azabicyclononyl, tetrahydropyranyl,
tetrahydropyronyl, tetrahydroquinoleinyl, tetrahydrothienyl and
dioxide thereof, dihydrothienyl dioxide, dioxindolyl, dioxinyl,
dioxenyl, dioxazinyl, thioxanyl, thioxolyl, thio-urazolyl,
thiotriazolyl, thiopyranyl, thiopyronyl, coumarinyl, quinoleinyl,
oxyquinoleinyl, quinuclidinyl, xanthinyl, dihydropyranyl,
benzodihydrofuryl, benzothiopyronyl, benzothiopyranyl,
benzoxazinyl, benzoxazolyl, benzodioxolyl, benzodioxanyl,
benzothiadiazolyl, benzotriazinyl, benzothiazolyl, benzoxazolyl,
phenothioxinyl, phenothiazolyl, phenothienyl, phenopyronyl,
phenoxazolyl, pyridinyl, dihydropyridinyl, tetrahydropyridinyl,
piperidinyl, morpholinyl, thiomorpholinyl, pyrazinyl, pyrimidinyl,
pyridazinyl, triazinyl, tetrazinyl, triazolyl, benzotriazolyl,
tetrazolyl, imidazolyl, pyrazolyl, thiazolyl, thiadiazolyl,
isothiazolyl, oxazolyl, oxadiazolyl, pyrrolyl, furyl, dihydrofuryl,
furoyl, hydantoinyl, dioxolanyl, dioxolyl, dithianyl, dithienyl,
dithiinyl, thienyl, indolyl, indazolyl, benzofuryl, benzothienyl,
quinolyl, quinazolinyl, quinoxalinyl, carbazolyl, phenoxazinyl,
phenothiazinyl, xanthenyl, purinyl, benzothienyl, naphtothienyl,
thianthrenyl, pyranyl, pyronyl, benzopyronyl, isobenzofuranyl,
chromenyl, phenoxathiinyl, indolizinyl, quinolizinyl, isoquinolyl,
phthalazinyl, naphthiridinyl, cinnolinyl, pteridinyl, carbolinyl,
acridinyl, perimidinyl, phenanthrolinyl, phenazinyl,
phenothiazinyl, imidazolinyl, imidazolidinyl, benzimidazolyl,
pyrazolinyl, pyrazolidinyl, pyrrolinyl, pyrrolidinyl, piperazinyl,
uridinyl, thymidinyl,cytidinyl, azirinyl, aziridinyl, diazirinyl,
diaziridinyl, oxiranyl, oxaziridinyl, dioxiranyl, thiiranyl,
azetyl, dihydroazetyl, azetidinyl, oxetyl, oxetanyl, thietyl,
thietanyl, oxetanonyl, diaza-bicyclooctyl, diazetyl,
diaziridinonyl, diaziridinethionyl, chromanyl, chroma-nonyl,
thiochromanyl, thiochromanonyl, thiochromenyl, benzofuranyl,
benzisothiazolyl, benzocarbazolyl, benzochromonyl,
benzisoalloxazinyl, benzocoumarinyl, thiocoumarinyl,
phenometoxazinyl, phenoparoxazinyl, phentriazinyl, thiodiazinyl,
thiodiazolyl, indoxyl, thioindoxyl, benzodiazinyl (e.g.
phtalazinyl), phtalidyl, phtalimidinyl, phtalazonyl, alloxazinyl,
dibenzo-pyronyl (i.e. xanthonyl), xanthionyl, isatyl, isopyrazolyl,
isopyrazolonyl, urazolyl, urazinyl, uretinyl, uretidinyl, succinyl,
succinimido, benzylsultimyl, and benzylsultamyl; [0027] R.sub.2 is
a radical selected from the group consisting of hydrogen; C.sub.1-5
alkyl; C.sub.2-7 alkenyl; aryl; alkylaryl; .omega.-hydroxy
C.sub.1-7 alkyl; .omega.-epoxy C.sub.1-7 alkyl; .omega.-carboxy
C.sub.1-7 alkyl (wherein the carboxy group may be acid, ester,
thioester, acid halide or amide); .omega.-cyano C.sub.1-7 alkyl;
arylalkyl; arylalkenyl; heterocyclic-substituted alkyl;
heterocyclic-substituted alkenyl; groups having the formula --S--R
(i.e. wherein a sulfur atom is attached to the nitrogen atom of the
pteridine ring) wherein R is a monovalent group selected from the
group consisting of C.sub.1-7 alkyl, aryl and C.sub.3-10 cycloalkyl
and wherein the said monovalent group is optionally substituted
with one or more substituents selected from the group consisting of
amino, amino-acid, alkylamino, arylamino, cycloalkylamino,
carboxylic acid, carboxylic ester, sulfonic acid and phosphonic
acid; and optionally substituted heterocyclic radicals preferably
selected from the group consisting of diazepinyl, oxadiazinyl,
thiadiazinyl, dithiazinyl, triazolonyl, diazepinonyl, triazepinyl,
triazepinonyl, tetrazepinonyl, benzoquinolinyl, benzothiazinyl,
benzothiazinonyl, benzoxathiinyl, benzodioxinyl, benzodithiinyl,
benzoxazepinyl, benzothiazepinyl, benzodiazepinyl, benzodioxepinyl,
benzodithiepinyl, benzoxazocinyl, benzothiazocinyl,
benzodiazocinyl, benzoxathiocinyl, benzodioxocinyl,
benzotrioxepinyl, benzoxathiazepinyl, benzoxadiazepinyl,
benzothiadiazepinyl, benzotri-azepinyl, benzoxathiepinyl,
benzotriazinonyl, benzoxazolinonyl, azetidinonyl, azaspiroundecyl,
dithiaspirodecyl, selenazinyl, selenazolyl, selenophenyl,
hypoxanthinyl, azahypoxanthinyl, bipyrazinyl, bipyridinyl,
oxazolidinyl, diselenopyrimidinyl, benzopyrenyl, benzopyranonyl,
benzophenazinyl, benzoquinolizinyl, dibenzocarbazolyl,
dibenzoacridinyl, dibenzophenazinyl, dibenzothiepinyl,
dibenzooxepinyl, dibenzopyranonyl, dibenzoquinoxalinyl,
dibenzothiazepinyl, dibenzoisoquinolinyl, tetraazaadamantyl,
thiatetraazaadamantyl, oxauracil, oxazinyl, dibenzothiophenyl,
dibenzofuranyl, oxazolinyl, oxazolonyl, azaindolyl, azolonyl,
thiazolinyl, thiazolonyl, thiazolidinyl, thiazanyl, pyrimidonyl,
thiopyrimidonyl, thiamorpholinyl, azlactonyl, naphtindazolyl,
naphtindolyl, naphtothiazolyl, naphtothioxolyl, naphtoxindolyl,
naphtotriazolyl, naphtopyranyl, indolinyl, indolizidinyl,
oxabicycloheptyl, azabenzimidazolyl, azacycloheptyl, azacyclooctyl,
azacyclononyl, azabicyclononyl, tetrahydropyranyl,
tetrahydropyronyl, tetrahydroquinoleinyl, tetrahydrothienyl and
dioxide thereof, dihydrothienyl dioxide, dioxindolyl, dioxinyl,
dioxenyl, dioxazinyl, thioxanyl, thioxolyl, thio-urazolyl,
thiotriazolyl, thiopyranyl, thiopyronyl, coumarinyl, quinoleinyl,
oxyquinoleinyl, quinuclidinyl, xanthinyl, dihydropyranyl,
benzodihydrofuryl, benzothiopyronyl, benzothiopyranyl,
benzoxazinyl, benzoxazolyl, benzodioxolyl, benzodioxanyl,
benzothiadiazolyl, benzotriazinyl, benzothiazolyl, benzoxazolyl,
phenothioxinyl, phenothiazolyl, phenothienyl, phenopyronyl,
phenoxazolyl, pyridinyl, dihydropyridinyl, tetrahydropyridinyl,
piperidinyl, morpholinyl, thiomorpholinyl, pyrazinyl, pyrimidinyl,
pyridazinyl, triazinyl, tetrazinyl, triazolyl, benzotriazolyl,
tetrazolyl, imidazolyl, pyrazolyl, thiazolyl, thiadiazolyl,
isothiazolyl, oxazolyl, oxadiazolyl, pyrrolyl, furyl, dihydrofuryl,
furoyl, hydantoinyl, dioxolanyl, dioxolyl, dithianyl, dithienyl,
dithiinyl, thienyl, indolyl, indazolyl, benzofuryl, benzothienyl,
quinolyl, quinazolinyl, quinoxalinyl, carbazolyl, phenoxazinyl,
phenothiazinyl, xanthenyl, purinyl, benzothienyl, naphtothienyl,
thianthrenyl, pyranyl, pyronyl, benzopyronyl, isobenzofuranyl,
chromenyl, phenoxathiinyl, indolizinyl, quinolizinyl, isoquinolyl,
phthalazinyl, naphthiridinyl, cinnolinyl, pteridinyl, carbolinyl,
acridinyl, perimidinyl, phenanthrolinyl, phenazinyl,
phenothiazinyl, imidazolinyl, imidazolidinyl, benzimidazolyl,
pyrazolinyl, pyrazolidinyl, pyrrolinyl, pyrrolidinyl, piperazinyl,
uridinyl, thymidinyl,cytidinyl, azirinyl, aziridinyl, diazirinyl,
diaziridinyl, oxiranyl, oxaziridinyl, dioxiranyl, thiiranyl,
azetyl, dihydroazetyl, azetidinyl, oxetyl, oxetanyl, thietyl,
thietanyl, oxetanonyl, diazabicyclooctyl, diazetyl, diaziridinonyl,
diaziridinethionyl, chromanyl, chromanonyl, thiochromanyl,
thiochromanonyl, thiochromenyl, benzofuranyl, benzisothiazolyl,
benzocarbazolyl, benzochromonyl, benzisoalloxazinyl,
benzocoumarinyl, thiocoumarinyl, phenometoxazinyl,
phenoparoxazinyl, phentriazinyl, thiodiazinyl, thiodiazolyl,
indoxyl, thioindoxyl, benzodiazinyl (e.g. phtalazinyl), phtalidyl,
phtalimidinyl, phtalazonyl, alloxazinyl, dibenzo-pyronyl (i.e.
xanthonyl), xanthionyl, isatyl, isopyrazolyl, isopyrazolonyl,
urazolyl, urazinyl, uretinyl, uretidinyl, succinyl, succinimido,
benzylsultimyl, and benzylsultamyl; [0028] at most one of R.sub.1
and R.sub.2 is hydrogen; and [0029] R.sub.4 is an atom or radical
selected from the group consisting of fluoro; iodo; C.sub.3-4
alkyl; C.sub.2-7 alkenyl; C.sub.2-7 alkynyl; halo C.sub.3-4 alkyl;
halo C.sub.1-2 alkyl wherein halo is fluoro or chloro; C.sub.1-4
alkoxy; C.sub.3-10 cycloalkoxy; aryloxy; arylalkyloxy;
oxyheterocyclic; heterocyclic-substituted alkyloxy; thio C.sub.1-7
alkyl; thio C.sub.3-10 cycloalkyl; thioaryl; thioheterocyclic;
arylalkylthio; heterocyclic-substituted alkylthio; hydroxylamino;
acetal; carboxylic acid esters, thioesters and amides;
thiocarboxylic acid; thiocarboxylic acid esters, thioesters and
amides; sulfhydryl; C.sub.2-7 alkylamino; cycloalkylamino;
alkenylamino; cycloalkenylamino; alkynylamino; arylamino;
arylalkylamino; hydroxyalkylamino; mercaptoalkyl-amino;
heterocyclic amino; heterocyclic-substituted alkylamino; oximino;
alkyloximino; hydrazino; alkylhydrazino; phenylhydrazino; cysteinyl
acid, esters or amides; aryl substituted with one or more
substituents selected from the group consisting of halogen, nitro,
C.sub.1-7 alkyl, C.sub.2-7 alkenyl, C.sub.2-7 alkynyl, halo
C.sub.1-7 alkyl, methoxy, C.sub.2-7 alkoxy, hydroxyl, sulfhydryl,
amino, C.sub.3-10 cycloalkoxy, aryloxy, arylalkyloxy,
oxyheterocyclic, heterocyclic-substituted alkyloxy, thio C.sub.1-7
alkyl, thio C.sub.3-10 cycloalkyl, thioaryl, thioheterocyclic,
arylalkylthio, heterocyclic-substituted alkylthio, formyl,
hydroxylamino, cyano, carboxylic acid or esters or thioesters or
amides thereof, thiocarboxylic acid or esters or thioesters or
amides thereof, C.sub.1-7 alkylamino, cycloalkylamino,
alkenylamino, cycloalkenylamino, alkynylamino, arylamino,
arylalkylamino, hydroxylalkylamino, mercaptoalkylamino,
heterocyclic amino, hydrazino, alkylhydrazino and phenylhydrazino;
optionally substituted heterocyclic radicals preferably other than
morpholino and piperidino, i.e. preferably selected from the group
consisting of diazepinyl, oxadiazinyl, thiadiazinyl, dithiazinyl,
triazolonyl, diazepinonyl, triazepinyl, triazepinonyl,
tetrazepinonyl, benzoquinolinyl, benzothiazinyl, benzothiazinonyl,
benzoxathiinyl, benzodi-oxinyl, benzodithiinyl, benzoxazepinyl,
benzothiazepinyl, benzodiazepinyl, benzodioxepinyl,
benzodithiepinyl, benzoxazocinyl, benzothiazocinyl,
benzodiazocinyl, benzoxathiocinyl, benzodioxocinyl,
benzotrioxepinyl, benzoxathiazepinyl, benzoxadiazepinyl,
benzothiadiazepinyl, benzotri-azepinyl, benzoxathiepinyl,
benzotriazinonyl, benzoxazolinonyl, azetidinonyl, azaspiroundecyl,
dithiaspirodecyl, selenazinyl, selenazolyl, selenophenyl,
hypoxanthinyl, azahypoxanthinyl, bipyrazinyl, bipyridinyl,
oxazolidinyl, diselenopyrimidinyl, benzopyrenyl, benzopyranonyl,
benzophenazinyl, benzoquinolizinyl, dibenzocarbazolyl,
dibenzoacridinyl, dibenzophenazinyl, dibenzothiepinyl,
dibenzooxepinyl, dibenzopyranonyl, dibenzoquinoxalinyl,
dibenzothiazepinyl, dibenzoisoquinolinyl, tetraazaadamantyl,
thiatetraaza-adamantyl, oxauracil, oxazinyl, dibenzothiophenyl,
dibenzofuranyl, oxazolinyl, oxazolonyl, azaindolyl, azolonyl,
thiazolinyl, thiazolonyl, thiazolidinyl, thiazanyl, pyrimidonyl,
thiopyrimidonyl, thiamorpholinyl, azlactonyl, naphtindazolyl,
naphtindolyl, naphtothiazolyl, naphtothioxolyl, naphtoxindolyl,
naphtotriazolyl, naphtopyranyl, indolinyl, indolizidinyl,
oxabicycloheptyl, azabenzimidazolyl, azacycloheptyl, azacyclooctyl,
azacyclononyl, azabicyclononyl, tetrahydrofuryl, tetrahydropyranyl,
tetrahydropyronyl, tetrahydroquinoleinyl, tetrahydrothienyl and
dioxide thereof, dihydrothienyl dioxide, dioxindolyl, dioxinyl,
dioxenyl, dioxazinyl, thioxanyl, thioxolyl, thio-urazolyl,
thiotriazolyl, thiopyranyl, thiopyronyl, coumarinyl, quinoleinyl,
oxyquinoleinyl, quinuclidinyl, xanthinyl, dihydropyranyl,
benzodihydrofuryl, benzothiopyronyl, benzothiopyranyl,
benzoxazinyl, benzoxazolyl, benzo-dioxolyl, benzodioxanyl,
benzothiadiazolyl, benzotriazinyl, benzothiazolyl, benzoxazolyl,
phenothioxinyl, phenothiazolyl, phenothienyl, phenopyronyl,
phenoxazolyl, pyridinyl, dihydropyridinyl, tetrahydropyridinyl,
piperidinyl, morpholinyl, thiomorpholinyl, pyrazinyl, pyrimidinyl,
pyridazinyl, triazinyl, tetrazinyl, triazolyl, benzotriazolyl,
imidazolyl, pyrazolyl, thiazolyl, thiadiazolyl, isothiazolyl,
oxazolyl, oxadiazolyl, pyrrolyl, furyl, dihydrofuryl, furoyl,
dioxolyl, dithienyl, dithiinyl, thienyl, indolyl, indazolyl,
benzofuryl, benzothienyl, quinolyl, quinazolinyl, quinoxalinyl,
carbazolyl, phenoxazinyl, phenothiazinyl, xanthenyl, purinyl,
benzothienyl, naphtothienyl, thianthrenyl, pyranyl, pyronyl,
benzopyronyl, isobenzofuranyl, chromenyl, phenoxathiinyl,
indolizinyl, quinolizinyl, isoquinolyl, phthalazinyl,
naphthiridinyl, cinnolinyl, pteridinyl, carbolinyl, acridinyl,
perimidinyl, phenanthrolinyl, phenazinyl, phenothiazinyl,
imidazolinyl, benzimidazolyl, pyrazolinyl, pyrazolidinyl,
pyrrolinyl, pyrrolidinyl, piperazinyl, uridinyl, thymidinyl,
cytidinyl, azirinyl, aziridinyl, diazirinyl, diaziridinyl,
oxiranyl, oxaziridinyl, dioxiranyl, thiiranyl, azetyl,
dihydroazetyl, azetidinyl, oxetyl, oxetanyl, thietyl, thietanyl,
oxetanonyl, diazabicyclooctyl, diazetyl, diaziridinonyl,
diaziridinethionyl, chromanyl, chromanonyl, thiochromanyl,
thiochromanonyl, thiochromenyl, benzofuranyl, benzisothiazolyl,
benzocarbazolyl, benzochromonyl, benzisoalloxazinyl,
benzocoumarinyl, thiocoumarinyl, phenometoxazinyl,
phenoparoxazinyl, phentriazinyl, thiodiazinyl, thiodiazolyl,
indoxyl, thioindoxyl, benzodiazinyl (e.g. phtalazinyl), phtalidyl,
phtalimidinyl, phtalazonyl, alloxazinyl, dibenzopyronyl (i.e.
xanthonyl), xanthionyl, isatyl, isopyrazolyl, isopyrazolonyl,
urazolyl, urazinyl, uretinyl, uretidinyl, succinyl, succinimido,
benzylsultimyl, and benzylsultamyl; aryl or heterocyclic radicals
substituted with an aliphatic spacer between the pteridine ring and
the aryl or heterocyclic radical, whereby said aliphatic spacer is
a branched or straight, saturated or unsaturated aliphatic chain of
2 to 4 carbon atoms which may contain one or more functions, atoms
or radicals selected from the group consisting of carbonyl (oxo),
alcohol (hydroxyl), ether, acetal, amino, imino, oximino,
alkyloximino, amino-acid, cyano, carboxylic acid or ester or
thioester or amide, nitro, thio C
.sub.1-7 alkyl, thio C.sub.3-10 cycloalkyl, C.sub.1-7 alkylamino,
cycloalkylamino, alkenylamino, cycloalkenylamino, alkynylamino,
arylamino, arylalkylamino, hydroxyalkylamino, mercaptoalkylamino,
heterocyclic amino, hydrazino, alkylhydrazino, phenylhydrazino,
sulfonyl, sulfonamido and halogen, or whereby said aliphatic spacer
is a methylene group containing a function, atom or radical chosen
from the group consisting of carbonyl (oxo), alcohol (hydroxyl),
ether, acetal, amino, imino, oximino, alkyloximino, amino-acid,
cyano, carboxylic acid or ester or thioester or amide, nitro, thio
C.sub.1-7 alkyl, thio C.sub.3-10 cycloalkyl, C.sub.1-7 alkylamino,
cycloalkylamino, alkenylamino, cycloalkenylamino, alkynylamino,
arylamino, arylalkylamino, hydroxyalkylamino, mercaptoalkylamino,
heterocyclic amino, hydrazino, alkylhydrazino, phenylhydrazino,
sulfonyl, sulfonamido, fluoro and chloro; branched or straight,
saturated or unsaturated aliphatic chain of 3 to 7 carbon atoms
containing one or more functions selected from the group consisting
of carbonyl (oxo), alcohol (hydroxyl), ether, acetal, amino, imino,
oximino, alkyloximino, aminoacid, cyano, carboxylic acid ester or
amide, nitro, thio C.sub.1-7 alkyl, thio C.sub.3-10 cycloalkyl,
C.sub.1-7 alkylamino, cycloalkylamino, alkenylamino,
cycloalkenylamino, alkynylamino, arylamino, arylalkylamino,
hydroxyalkylamino, mercaptoalkylamino, heterocyclic amino,
hydrazino, alkylhydrazino, phenylhydrazino, sulfonyl, sulfonamido
and halogen; hydroxyethyl; oximinoethyl; alkyloximinoethyl; and
methyl or ethyl or ethenyl containing one or more functions, atoms
or radicals selected from the group consisting of ether, acetal,
amino, imino, amino-acid, cyano, carboxylic acid or ester or
thioester or amide, nitro, thio C.sub.1-7 alkyl, thio C.sub.3-10
cycloalkyl, C.sub.1-7 alkylamino, cycloalkylamino, alkenylamino,
cycloalkenyl-amino, alkynylamino, arylalkylamino,
hydroxyalkylamino, mercaptoalkylamino, heterocyclic amino,
hydrazino, alkylhydrazino, phenylhydrazino, sulfonyl, sulfonamido,
fluoro and chloro; [0030] c) if one or more of Y.sub.1 and Y.sub.2
is sulfur and at most one of Y.sub.1 and Y.sub.2 is oxygen, then:
[0031] each of R.sub.1 and R.sub.2 is a radical independently
selected from the group consisting of hydrogen; C.sub.1-7 alkyl;
C.sub.2-7 alkenyl; aryl; alkylaryl; .omega.-hydroxy C.sub.1-7
alkyl; .omega.-epoxy C.sub.1-7 alkyl; .omega.-carboxy C.sub.1-7
alkyl (wherein the carboxy group may be acid, ester, thioester,
acid halide or amide); .omega.-cyano C.sub.1-7 alkyl; arylalkyl;
arylalkenyl; heterocyclic-substituted alkyl;
heterocyclic-substituted alkenyl; groups having the formula --S--R
(i.e. wherein a sulfur atom is attached to the nitrogen atom of the
pteridine ring) wherein R is a monovalent group selected from the
group consisting of C.sub.1-7 alkyl, aryl and C.sub.3-10 cycloalkyl
and wherein the said monovalent group is optionally substituted
with one or more substituents selected from the group consisting of
amino, amino-acid, alkylamino, arylamino, cycloalkylamino,
carboxylic acid, carboxylic ester, sulfonic acid and phosphonic
acid; and optionally substituted heterocyclic radicals; [0032] each
of R.sub.3 and R.sub.4 is an atom or radical independently selected
from the group consisting of hydrogen; halogen; C.sub.2-4 alkyl;
C.sub.2-7 alkenyl; C.sub.2-7 alkynyl; halo C.sub.1-4 alkyl;
C.sub.2-4 alkoxy; C.sub.3-10 cycloalkoxy; aryloxy; arylalkyloxy;
oxyheterocyclic; heterocyclic-substituted alkyloxy; thio C.sub.2-7
alkyl; thio C.sub.3-10 cycloalkyl; thioaryl; thioheterocyclic;
arylalkylthio; heterocyclic-substituted alkylthio; hydroxylamino;
acetal; formyl; cyano; carboxylic acid; carboxylic acid esters,
thioesters and amides; thiocarboxylic acid; thiocarboxylic acid
esters, thioesters and amides; amino; alkylamino; cycloalkylamino;
alkenylamino; cycloalkenylamino; alkynylamino; arylamino;
arylalkylamino; hydroxyalkylamino; mercaptoalkylamino; heterocyclic
amino; heterocyclic-substituted alkylamino; oximino; alkyloximino;
hydrazino; alkylhydrazino; phenylhydrazino; cysteinyl acid, esters
or amides; aryl substituted with one or more substituents selected
from the group consisting of halogen, C.sub.1-4 alkyl, C.sub.2-7
alkenyl, C.sub.2-7 alkynyl, halo C.sub.1-4 alkyl, C.sub.1-4 alkoxy,
hydroxyl, sulfhydryl, amino, C.sub.3-10 cycloalkoxy, aryloxy,
arylalkyloxy, oxyheterocyclic, heterocyclic-substituted alkyloxy,
thio C.sub.1-7 alkyl, thio C.sub.3-10 cycloalkyl, thioaryl,
thioheterocyclic, arylalkylthio, heterocyclic-substituted
alkylthio, formyl, hydroxylamino, cyano, carboxylic acid or esters
or thioesters or amides thereof, thiocarboxylic acid or esters or
thioesters or amides thereof, C.sub.1-7 alkylamino,
cycloalkylamino, alkenylamino, cycloalkenylamino, alkynylamino,
arylamino, arylalkylamino, hydroxylalkylamino, mercaptoalkylamino,
heterocyclic amino, hydrazino, alkylhydrazino and phenylhydrazino;
optionally substituted heterocyclic radicals; aryl or heterocyclic
radicals substituted with an aliphatic spacer (linking group)
between the pteridine ring and the aryl or heterocyclic radical,
whereby said aliphatic spacer is a branched or straight, saturated
or unsaturated aliphatic chain of 1 to 4 carbon atoms (such as a
C.sub.1-4 alkylene) which may contain one or more functions, atoms
or radicals selected from the group consisting of carbonyl (oxo),
alcohol (hydroxyl), ether, acetal, amino, imino, oximino,
alkyloximino, amino-acid, cyano, carboxylic acid or ester or
thioester or amide, nitro, thio C.sub.1-7 alkyl, thio C.sub.3-10
cycloalkyl, C.sub.1-7 alkylamino, cycloalkylamino, alkenylamino,
cycloalkenylamino, alkynylamino, arylamino, arylalkylamino,
hydroxyalkylamino, mercaptoalkyl-amino, heterocyclic amino,
hydrazino, alkylhydrazino, phenylhydrazino, sulfonyl, sulfonamido
and halogen; branched or straight, saturated or unsaturated
aliphatic chain of 1 to 7 carbon atoms containing one or more
functions selected from the group consisting of carbonyl (oxo),
alcohol (hydroxyl), ether, acetal, amino, imino, oximino,
alkyloximino, amino-acid, cyano, carboxylic acid ester or amide,
nitro, thio C.sub.1-7 alkyl, thio C.sub.3-10 cycloalkyl, C.sub.1-7
alkylamino, cycloalkylamino, alkenylamino, cycloalkenylamino,
alkynylamino, arylamino, arylalkylamino, hydroxylalkylamino,
mercaptoalkyl-amino, heterocyclic amino, hydrazino, alkylhydrazino,
phenylhydrazino, sulfonyl, sulfonamido and halogen; or R.sub.4 and
R.sub.3 together form an aryl radical being optionally substituted
with one or more substituents R.sub.a each independently selected
from the group consisting of amino, alkylamino, cycloalkylamino,
alkenylamino, cycloalkenylamino, alkynylamino, arylamino,
arylalkylamino, hydroxyalkylamino, mercaptoalkylamino, heterocyclic
amino and heterocyclic-substituted alkylamino, wherein each
substituent R.sub.a may further comprise one or more functions
selected from the group consisting of carbonyl, amino and carboxyl,
and wherein two adjacent substituents R.sub.a may together form an
heterocyclic radical; and [0033] at most one of R.sub.1, R.sub.2,
R.sub.3 and R.sub.4 is hydrogen; [0034] d) if Y.sub.1 and Y.sub.2
are both oxygen and none of R.sub.3 and R.sub.4 is hydrogen, then:
[0035] R.sub.2 is a radical selected from the group consisting of
C.sub.1-7 alkyl; C.sub.2-7 alkenyl; aryl; alkylaryl;
.omega.-hydroxy C.sub.1-7 alkyl; .omega.-epoxy C.sub.1-7 alkyl;
.omega.-carboxy C.sub.1-7 alkyl (wherein the carboxy group may be
acid, ester, thioester, acid halide or amide); .omega.-cyano
C.sub.1-7 alkyl; arylalkyl; arylalkenyl; heterocyclic-substituted
alkyl; heterocyclic-substituted alkenyl; groups having the formula
--S--R (i.e. wherein a sulfur atom is attached to the nitrogen atom
of the pteridine ring) wherein R is a monovalent group selected
from the group consisting of C.sub.1-7 alkyl, aryl and C.sub.3-10
cycloalkyl and wherein the said monovalent group is optionally
substituted with one or more substituents selected from the group
consisting of amino, amino-acid, alkylamino, arylamino,
cycloalkylamino, carboxylic acid, carboxylic ester, sulfonic acid
and phosphonic acid; and optionally substituted heterocyclic
radicals; [0036] R.sub.1 is an atom or radical independently
defined as R.sub.2, or is hydrogen; [0037] R.sub.4 is an atom or
radical selected from the group consisting of halogen (preferably
chloro); C.sub.2-7 alkenyl; C.sub.2-7 alkynyl; C.sub.2-7 haloalkyl;
fluoromethyl; C.sub.2-4 alkoxy; C.sub.3-10 cycloalkoxy; aryloxy;
arylalkyloxy; oxyheterocyclic; heterocyclic-substituted alkyloxy;
thio C.sub.1-7 alkyl; thio C.sub.3-10 cycloalkyl; thioaryl;
thioheterocyclic; arylalkylthio; heterocyclic-substituted
alkylthio; hydroxyl-amino; acetal; carboxylic acid thioesters and
amides; thiocarboxylic acid; thiocarboxylic acid esters, thioesters
and amides; sulfhydryl; C.sub.2-7 alkylamino; cycloalkylamino;
alkenylamino; cycloalkenylamino; alkynylamino; arylamino;
arylalkylamino; hydroxyalkylamino; mercaptoalkylamino; heterocyclic
amino; heterocyclic-substituted alkylamino; hydrazino;
alkylhydrazino; phenylhydrazino; cysteinyl acid, esters or amides;
aryl optionally substituted with one or more substituents selected
from the group consisting of halogen, C.sub.1-4 alkyl, C.sub.2-7
alkenyl, C.sub.2-7 alkynyl, halo C.sub.1-7 alkyl, C.sub.1-7 alkoxy,
hydroxyl, sulfhydryl, amino, C.sub.3-10 cycloalkoxy, aryloxy,
arylalkyloxy, oxyheterocyclic, heterocyclic-substituted alkyloxy,
thio C.sub.1-7 alkyl, thio C.sub.3-10 cycloalkyl, thioaryl,
thioheterocyclic, arylalkylthio, heterocyclic-substituted
alkylthio, formyl, hydroxylamino, cyano, carboxylic acid or esters
or thioesters or amides thereof, thiocarboxylic acid or esters or
thioesters or amides thereof, C.sub.1-7 alkylamino,
cycloalkylamino, alkenylamino, cycloalkenylamino, alkynylamino,
arylamino, arylalkylamino, hydroxylalkylamino, mercaptoalkylamino,
heterocyclic amino, hydrazino, alkylhydrazino and phenylhydrazino;
optionally substituted heterocyclic radicals preferably selected
from the group consisting of diazepinyl, oxadiazinyl, thiadiazinyl,
dithiazinyl, triazolonyl, diazepinonyl, triazepinyl, triazepinonyl,
tetrazepinonyl, benzoquinolinyl, benzothiazinyl, benzothiazinonyl,
benzoxathiinyl, benzodioxinyl, benzodithiinyl, benzoxaze-pinyl,
benzothiazepinyl, benzodiazepinyl, benzodioxepinyl,
benzodithiepinyl, benzoxazocinyl, benzothiazocinyl,
benzodiazocinyl, benzoxathiocinyl, benzodioxocinyl,
benzotrioxepinyl, benzoxathiazepinyl, benzoxadiazepinyl,
benzothiadiazepinyl, benzotriazepinyl, benzoxathiepinyl,
benzotriazinonyl, benzoxazolinonyl, azetidinonyl, azaspiroundecyl,
dithiaspirodecyl, selenazinyl, selenazolyl, selenophenyl,
hypoxanthinyl, azahypoxanthinyl, bipyrazinyl, bipyridinyl,
oxazolidinyl, diselenopyrimidinyl, benzopyrenyl, benzopyranonyl,
benzophenazinyl, benzoquinolizinyl, dibenzocarbazolyl,
dibenzoacridinyl, dibenzophenazinyl, dibenzothiepinyl,
dibenzooxepinyl, dibenzopyranonyl, dibenzoquinoxalinyl,
dibenzothiazepinyl, dibenzoiso-quinolinyl, tetraazaadamantyl,
thiatetraazaadamantyl, oxauracil, oxazinyl, dibenzothiophenyl,
dibenzofuranyl, oxazolinyl, oxazolonyl, azaindolyl, azolonyl,
thiazolinyl, thiazolonyl, thiazolidinyl, thiazanyl, pyrimidonyl,
thiopyrimidonyl, thiamorpholinyl, azlactonyl, naphtindazolyl,
naphtindolyl, naphtothiazolyl, naphtothioxolyl, naphtoxindolyl,
naphtotriazolyl, naphtopyranyl, indolinyl, indolizidinyl,
tetrahydrofuryl, oxabicycloheptyl, azabenzimidazolyl,
azacycloheptyl, azacyclooctyl, azacyclononyl, azabicyclononyl,
tetrahydropyranyl, tetrahydropyronyl, tetrahydroquinoleinyl,
tetrahydrothienyl and dioxide thereof, dihydrothienyl dioxide,
dioxindolyl, dioxinyl, dioxenyl, dioxazinyl, thioxanyl, thioxolyl,
thio-urazolyl, thiotriazolyl, thiopyranyl, thiopyronyl, coumarinyl,
quinoleinyl, oxyquinoleinyl, quinuclidinyl, xanthinyl,
dihydropyranyl, benzodihydrofuryl, benzothiopyronyl,
benzothiopyranyl, benzoxazinyl, benzoxazolyl, benzodioxolyl,
benzodioxanyl, benzothiadiazolyl, benzotriazinyl, benzothiazolyl,
benzoxazolyl, phenothioxinyl, phenothiazolyl, phenothienyl,
phenopyronyl, phenoxazolyl, pyridinyl, dihydropyridinyl,
tetrahydropyridinyl, piperidinyl, morpholinyl, thiomorpholinyl,
pyrazinyl, pyrimidinyl, pyridazinyl, triazinyl, tetrazinyl,
triazolyl, benzotriazolyl, tetrazolyl, imidazolyl, pyrazolyl,
thiazolyl, thiadiazolyl, isothiazolyl, oxazolyl, oxadiazolyl,
pyrrolyl, furyl, dihydrofuryl, furoyl, hydantoinyl, dioxolyl,
dithianyl, dithienyl, dithiinyl, thienyl, indolyl, indazolyl,
benzofuryl, benzothienyl, quinolyl, quinazolinyl, quinoxalinyl,
carbazolyl, phenoxazinyl, phenothiazinyl, xanthenyl, purinyl,
benzothienyl, naphtothienyl, thianthrenyl, pyranyl, pyronyl,
benzopyronyl, isobenzofuranyl, chromenyl, phenoxathiinyl,
indolizinyl, quinolizinyl, isoquinolyl, phthalazinyl,
naphthiridinyl, cinnolinyl, pteridinyl, carbolinyl, acridinyl,
perimidinyl, phenanthrolinyl, phenazinyl, phenothiazinyl,
imidazolinyl, imidazolidinyl, benzimidazolyl, pyrazolinyl,
pyrazolidinyl, pyrrolinyl, pyrrolidinyl, piperazinyl, uridinyl,
thymidinyl,cytidinyl, azirinyl, aziridinyl, diazirinyl,
diaziridinyl, oxiranyl, oxaziridinyl, dioxiranyl, thiiranyl,
azetyl, dihydroazetyl, azetidinyl, oxetyl, oxetanyl, thietyl,
thietanyl, oxetanonyl, diazabicyclooctyl, diazetyl, diaziridinonyl,
diaziridinethionyl, chromanyl, chromanonyl, thiochromanyl,
thiochromanonyl, thiochromenyl, benzofuranyl, benzisothiazolyl,
benzocarbazolyl, benzochromonyl, benzisoalloxazinyl,
benzocoumarinyl, thiocoumarinyl, phenometoxazinyl,
phenoparoxazinyl, phentriazinyl, thiodiazinyl, thiodiazolyl,
indoxyl, thioindoxyl, benzodiazinyl (e.g. phtalazinyl), phtalidyl,
phtalimidinyl, phtalazonyl, alloxazinyl, dibenzopyronyl (i.e.
xanthonyl), xanthionyl, isatyl, isopyrazolyl, isopyrazolonyl,
urazolyl, urazinyl, uretinyl, uretidinyl, succinyl, succinimido,
benzylsultimyl, and benzylsultamyl; aryl or heterocyclic radicals
substituted with an aliphatic spacer between the pteridine ring and
the aryl or heterocyclic radical, whereby said aliphatic spacer is
a branched or straight, saturated or unsaturated aliphatic chain of
1 to 4 carbon atoms which may contain one or more functions, atoms
or radicals selected from the group consisting of carbonyl (oxo),
alcohol (hydroxyl), ether, acetal, amino, imino, oximino,
alkyloximino, amino-acid, cyano, carboxylic acid or ester or
thioester or amide, nitro, thio C.sub.1-7 alkyl, thio C.sub.3-10
cycloalkyl, C.sub.1-7 alkylamino, cycloalkylamino, alkenylamino,
cycloalkenylamino, alkynylamino, arylamino, arylalkylamino,
hydroxyalkylamino, mercaptoalkylamino, heterocyclic amino,
hydrazino, alkylhydrazino, phenylhydrazino, sulfonyl, sulfonamido
and halogen; branched or straight, saturated or unsaturated
aliphatic chain of 2 to 7 carbon atoms containing one or more
atoms, functions or radicals selected from the group consisting of
carbonyl (oxo), alcohol (hydroxyl), ether, acetal, amino, imino,
oximino, alkyloximino, amino-acid, cyano, carboxylic acid ester or
amide, nitro, thio C
.sub.1-7 alkyl, thio C.sub.3-10 cycloalkyl, C.sub.1-7 alkylamino,
cycloalkylamino, alkenylamino, cycloalkenylamino, alkynylamino,
arylamino, arylalkylamino, hydroxylalkylamino, mercaptoalkylamino,
heterocyclic amino, hydrazino, alkylhydrazino, phenylhydrazino,
sulfonyl, sulfonamido and halogen; hydroxyethyl; and [0038] R.sub.3
is an atom or radical independently defined as R.sub.4, or is amino
or methoxy, or R.sub.4 and R.sub.3 together form an aryl radical
being optionally substituted with one or more substituents R.sub.a
each independently selected from the group consisting of amino,
alkylamino, cycloalkylamino, alkenylamino, cycloalkenylamino,
alkynylamino, arylamino, arylalkylamino, hydroxyalkylamino,
mercaptoalkylamino, heterocyclic amino and heterocyclic-substituted
alkylamino, wherein each substituent R.sub.a may further comprise
one or more functions selected from the group consisting of
carbonyl, amino and carboxyl, and wherein two adjacent substituents
R.sub.a may together form an heterocyclic radical; [0039] e) if
Y.sub.1 and Y.sub.2 are both oxygen and none of R.sub.3 and R.sub.4
is hydrogen, then: [0040] R.sub.1 is a radical selected from the
group consisting of C.sub.2-7 alkyl; C.sub.2-7 alkenyl; aryl;
alkylaryl; .omega.-hydroxy C.sub.1-7 alkyl; .omega.-epoxy C.sub.1-7
alkyl; .omega.-carboxy C.sub.1-7 alkyl (wherein the carboxy group
may be acid, ester, thioester, acid halide or amide); .omega.-cyano
C.sub.1-7 alkyl; arylalkyl; arylalkenyl; heterocyclic-substituted
alkyl; heterocyclic-substituted alkenyl; groups having the formula
--S--R (i.e. wherein a sulfur atom is attached to the nitrogen atom
of the pteridine ring) wherein R is a monovalent group selected
from the group consisting of C.sub.1-7 alkyl, aryl and C.sub.3-10
cycloalkyl and wherein the said monovalent group is optionally
substituted with one or more substituents selected from the group
consisting of amino, amino-acid, alkylamino, arylamino,
cycloalkylamino, carboxylic acid, carboxylic ester, sulfonic acid
and phosphonic acid; and optionally substituted heterocyclic
radicals; [0041] R.sub.2 is hydrogen; [0042] R.sub.3 is an atom or
radical selected from the group consisting of chloro, fluoro;
bromo; iodo; C.sub.2-7 alkyl; C.sub.2-7 alkenyl; C.sub.2-7 alkynyl;
C.sub.2-7 haloalkyl; C.sub.1-4 alkoxy; C.sub.3-10 cycloalkoxy;
aryloxy; arylalkyloxy; oxyheterocyclic; heterocyclic-substituted
alkyloxy; thio C.sub.1-7 alkyl; thio C.sub.3-10 cycloalkyl;
thioaryl; thioheterocyclic; arylalkylthio; heterocyclic-substituted
alkylthio; hydroxyl-amino; acetal; carboxylic acid esters,
thioesters and amides; thiocarboxylic acid; thiocarboxylic acid
esters, thioesters and amides; sulfhydryl; amino; alkylamino;
cycloalkylamino; alkenylamino; cycloalkenyl-amino; alkynylamino;
arylamino; arylalkylamino; hydroxyalkylamino; mercaptoalkylamino;
heterocyclic amino; heterocyclic-substituted alkylamino; hydrazino;
alkylhydrazino; phenylhydrazino; cysteinyl acid, esters or amides;
aryl substituted with one or more substituents selected from the
group consisting of halogen, C.sub.1-4 alkyl, C.sub.2-7 alkenyl,
C.sub.2-7 alkynyl, C.sub.1-4 haloalkyl, C.sub.1-4 alkoxy, hydroxyl,
sulfhydryl, amino, C.sub.3-10 cycloalkoxy, aryloxy, arylalkyloxy,
oxyheterocyclic, heterocyclic-substituted alkyloxy, thio C.sub.1-7
alkyl, thio C.sub.3-10 cycloalkyl, thioaryl, thioheterocyclic,
arylalkylthio, heterocyclic-substituted alkylthio, formyl,
hydroxylamino, cyano, carboxylic acid or esters or thioesters or
amides thereof, thiocarboxylic acid or esters or thioesters or
amides thereof, C.sub.1-7 alkylamino, cycloalkylamino,
alkenylamino, cycloalkenylamino, alkynylamino, arylamino,
arylalkylamino, hydroxyalkylamino, mercaptoalkylamino, heterocyclic
amino, hydrazino, alkylhydrazino and phenylhydrazino; optionally
substituted heterocyclic radicals preferably selected from the
group consisting of diazepinyl, oxadiazinyl, thiadiazinyl,
dithiazinyl, triazolonyl, diazepinonyl, triazepinyl, triazepinonyl,
tetrazepinonyl, benzoquinolinyl, benzothiazinyl, benzothiazinonyl,
benzoxathiinyl, benzodioxinyl, benzodithiinyl, benzoxazepinyl,
benzothiazepinyl, benzodiazepinyl, benzodioxepinyl,
benzodithiepinyl, benzoxazocinyl, benzothiazocinyl,
benzodiazocinyl, benzoxathiocinyl, benzodioxocinyl,
benzotrioxepinyl, benzoxathiazepinyl, benzoxadiazepinyl,
benzothiadiazepinyl, benzotriazepinyl, benzoxathiepinyl,
benzotriazinonyl, benzoxazolinonyl, azetidinonyl, azaspiroundecyl,
dithiaspirodecyl, selenazinyl, selenazolyl, selenophenyl,
hypoxanthinyl, azahypoxanthinyl, bipyrazinyl, bipyridinyl,
oxazolidinyl, diselenopyrimidinyl, benzopyrenyl, benzopyranonyl,
benzophenazinyl, benzoquinolizinyl, dibenzocarbazolyl,
dibenzoacridinyl, dibenzophenazinyl, dibenzothiepinyl,
dibenzooxepinyl, dibenzopyranonyl, dibenzoquinoxalinyl,
dibenzothiazepinyl, dibenzoisoquinolinyl, tetraazaadamantyl,
thiatetraazaadamantyl, oxauracil, oxazinyl, dibenzothiophenyl,
dibenzofuranyl, oxazolinyl, oxazolonyl, azaindolyl, azolonyl,
thiazolinyl, thiazolonyl, thiazolidinyl, thiazanyl, pyrimidonyl,
thiopyrimidonyl, thiamorpholinyl, azlactonyl, naphtindazolyl,
naphtindolyl, naphtothiazolyl, naphtothioxolyl, naphtoxindolyl,
naphtotriazolyl, naphtopyranyl, indolinyl, indolizidinyl,
tetrahydrofuryl, oxabicycloheptyl, azabenzimidazolyl,
azacycloheptyl, azacyclooctyl, azacyclononyl, azabicyclononyl,
tetrahydropyranyl, tetrahydropyronyl, tetrahydroquinoleinyl,
tetrahydrothienyl and dioxide thereof, dihydrothienyl dioxide,
dioxindolyl, dioxinyl, dioxenyl, dioxazinyl, thioxanyl, thioxolyl,
thio-urazolyl, thiotriazolyl, thiopyranyl, thiopyronyl, coumarinyl,
quinoleinyl, oxyquinoleinyl, quinuclidinyl, xanthinyl,
dihydropyranyl, benzodihydrofuryl, benzothiopyronyl,
benzothiopyranyl, benzoxazinyl, benzoxazolyl, benzodioxolyl,
benzodioxanyl, benzothiadiazolyl, benzotriazinyl, benzothiazolyl,
benzoxazolyl, phenothioxinyl, phenothiazolyl, phenothienyl,
phenopyronyl, phenoxazolyl, pyridinyl, dihydropyridinyl,
tetrahydropyridinyl, piperidinyl, morpholinyl, thiomorpholinyl,
pyrazinyl, pyrimidinyl, pyridazinyl, triazinyl, tetrazinyl,
triazolyl, benzotriazolyl, tetrazolyl, imidazolyl, pyrazolyl,
thiazolyl, thiadiazolyl, isothiazolyl, oxazolyl, oxadiazolyl,
pyrrolyl, furyl, dihydrofuryl, furoyl, hydantoinyl, dioxolyl,
dithianyl, dithienyl, dithiinyl, thienyl, indolyl, indazolyl,
benzofuryl, benzothienyl, quinolyl, quinazolinyl, quinoxalinyl,
carbazolyl, phenoxazinyl, phenothiazinyl, xanthenyl, purinyl,
benzothienyl, naphtothienyl, thianthrenyl, pyranyl, pyronyl,
benzopyronyl, isobenzofuranyl, chromenyl, phenoxathiinyl,
indolizinyl, quinolizinyl, isoquinolyl, phthalazinyl,
naphthiridinyl, cinnolinyl, pteridinyl, carbolinyl, acridinyl,
perimidinyl, phenanthrolinyl, phenazinyl, phenothiazinyl,
imidazolinyl, imidazolidinyl, benzimidazolyl, pyrazolinyl,
pyrazolidinyl, pyrrolinyl, pyrrolidinyl, piperazinyl, uridinyl,
thymidinyl,cytidinyl, azirinyl, aziridinyl, diazirinyl,
diaziridinyl, oxiranyl, oxaziridinyl, dioxiranyl, thiiranyl,
azetyl, dihydroazetyl, azetidinyl, oxetyl, oxetanyl, thietyl,
thietanyl, oxetanonyl, diazabicyclooctyl, diazetyl, diaziridinonyl,
diaziridinethionyl, chromanyl, chroma-nonyl, thiochromanyl,
thiochromanonyl, thiochromenyl, benzofuranyl, benzisothiazolyl,
benzocarbazolyl, benzochromonyl, benzisoalloxazinyl,
benzocoumarinyl, thiocoumarinyl, phenometoxazinyl,
phenoparoxazinyl, phentriazinyl, thiodiazinyl, thiodiazolyl,
indoxyl, thioindoxyl, benzodiazinyl (e.g. phtalazinyl), phtalidyl,
phtalimidinyl, phtalazonyl, alloxazinyl, dibenzopyronyl (i.e.
xanthonyl), xanthionyl, isatyl, isopyrazolyl, isopyrazolonyl,
urazolyl, urazinyl, uretinyl, uretidinyl, succinyl, succinimido,
benzylsultimyl, and benzylsultamyl; aryl or heterocyclic radicals
substituted with an aliphatic spacer between the pteridine ring and
the aryl or heterocyclic radical, whereby said aliphatic spacer is
a branched or straight, saturated or unsaturated aliphatic chain of
1 to 4 carbon atoms (e.g. C.sub.1-4 alkylene) which may contain one
or more functions, atoms or radicals selected from the group
consisting of carbonyl (oxo), alcohol (hydroxyl), ether, acetal,
amino, imino, oximino, alkyloximino, amino-acid, cyano, carboxylic
acid or ester or thioester or amide, nitro, thio C.sub.1-7 alkyl,
thio C.sub.3-10 cycloalkyl, C.sub.1-7 alkylamino, cycloalkylamino,
alkenylamino, cycloalkenylamino, alkynylamino, arylamino,
arylalkylamino, hydroxyalkylamino, mercaptoalkylamino, heterocyclic
amino, hydrazino, alkylhydrazino, phenylhydrazino, sulfonyl,
sulfonamido and halogen; branched or straight, saturated or
unsaturated aliphatic chain of 1 to 7 carbon atoms containing one
or more atoms, functions or radicals selected from the group
consisting of carbonyl (oxo), alcohol (hydroxyl), ether, acetal,
amino, imino, oximino, alkyloximino, amino-acid, cyano, carboxylic
acid ester or amide, nitro, thio C.sub.1-7 alkyl, thio C.sub.3-10
cycloalkyl, C.sub.1-7 alkylamino, cycloalkylamino, alkenylamino,
cycloalkenylamino, alkynylamino, arylamino, arylalkylamino,
hydroxylalkylamino, mercaptoalkyl-amino, heterocyclic amino,
hydrazino, alkylhydrazino, phenylhydrazino, sulfonyl, sulfonamido
and halogen; hydroxyethyl; and [0043] R.sub.4 is an atom or radical
independently defined as R.sub.3, or is chloro, or R.sub.4 and
R.sub.3 together form an aryl radical being optionally substituted
with one or more substituents R.sub.a each independently selected
from the group consisting of amino, alkylamino, cycloalkylamino,
alkenylamino, cycloalkenylamino, alkynylamino, arylamino,
arylalkylamino, hydroxyalkylamino, mercaptoalkylamino, heterocyclic
amino and heterocyclic-substituted alkylamino, wherein each
substituent R.sub.a may further comprise one or more functions
selected from the group consisting of carbonyl, amino and carboxyl,
and wherein two adjacent substituents R.sub.a may together form an
heterocyclic radical; [0044] f) if Y.sub.1 and Y.sub.2 are both
oxygen and R.sub.2 and R.sub.3 are both hydrogen, then: [0045]
R.sub.1 is a radical selected from the group consisting of
C.sub.1-7 alkyl; C.sub.2-7 alkenyl; aryl; alkylaryl;
.omega.-hydroxy C.sub.1-7 alkyl; .omega.-epoxy C.sub.1-7 alkyl;
.omega.-carboxy C.sub.1-7 alkyl (wherein the carboxy group may be
acid, ester, thioester, acid halide or amide); .omega.-cyano
C.sub.1-7 alkyl; arylalkyl; arylalkenyl; heterocyclic-substituted
alkyl; heterocyclic-substituted alkenyl; groups having the formula
--S--R (i.e. wherein a sulfur atom is attached to the nitrogen atom
of the pteridine ring) wherein R is a monovalent group selected
from the group consisting of C.sub.1-7 alkyl, aryl and C.sub.3-10
cycloalkyl and wherein the said monovalent group is optionally
substituted with one or more substituents selected from the group
consisting of amino, amino-acid, alkylamino, arylamino,
cycloalkylamino, carboxylic acid, carboxylic ester, sulfonic acid
and phosphonic acid; and optionally substituted heterocyclic
radicals; [0046] R.sub.4 is an atom or a radical selected from the
group consisting of halogen (preferably chloro), cyano, amino,
alkylamino, cycloalkylamino, alkenylamino, cycloalkenylamino,
alkynylamino, arylamino, arylalkylamino, hydroxyalkylamino,
mercaptoalkylamino, heterocyclic amino and heterocyclic-substituted
alkylamino; or a pharmaceutically acceptable salt or an enantiomer
thereof.
[0047] Within the scope of this invention, the term "substituted
heterocyclic radicals" include, but are not limited to, such groups
represented by the following general formula (II): ##STR3##
wherein: ##STR4## schematically represents a saturated or partly
unsaturated heterocyclic ring with at least two nitrogen atoms in
the said heterocyclic ring and with a total of 5 to 7 atoms in the
said heterocyclic ring, and optionally with one or more other
heteroatoms (e.g. oxygen or sulfur) in the said heterocyclic ring
or attached to one or more carbon atoms of said heterocyclic ring
(for instance in the form of a carbonyl or thiocarbonyl group),
wherein one of said at least two nitrogen atoms in the heterocyclic
ring is attached to a carbon atom of the pteridine ring at any of
positions 1, 3, 6 or 7 of the pteridine ring, wherein the said
heterocyclic ring may be fused to one or more aromatic hydrocarbon
rings, and wherein: [0048] each substituent R.sub.0 of the
heterocyclic ring (III) is a group independently selected from the
group consisting of halogen, nitro, C.sub.1-7 alkyl (optionally
containing one or more functions or radicals selected from the
group consisting of halogen, carbonyl, thiocarbonyl, hydroxyl,
sulfhydryl, C.sub.1-7 alkoxy, thio C.sub.1-7 alkyl, thio C.sub.3-10
cycloalkyl, acetal, thioacetal, imino, oximino, alkyloximino,
amino-acid, cyano, (thio)carboxylic acid, (thio)carboxylic acid
ester or amide, nitro, amino, C.sub.1-7 alkylamino,
cycloalkylamino, alkenylamino, cycloalkenylamino, alkynyl-amino,
arylamino, arylalkylamino, hydroxyalkylamino, mercaptoalkylamino,
heterocyclic amino, hydrazino, alkylhydrazino, phenylhydrazino,
sulfonyl and sulfonamido), C.sub.2-7 alkenyl, C.sub.2-7 alkynyl,
halo C.sub.1-7 alkyl, C.sub.3-10 cycloalkyl, aryl, arylalkyl,
alkylaryl, alkylacyl, arylacyl, hydroxyl, sulfhydryl, amino,
C.sub.1-7 alkylamino, cycloalkylamino, alkenylamino,
cycloalkenylamino, alkynylamino, arylamino, arylalkylamino,
hydroxyalkylamino, mercaptoalkylamino, heterocyclic-amino,
hydrazino, alkyl-hydrazino, phenylhydrazino, C.sub.1-7 alkoxy,
C.sub.3-10 cycloalkoxy, aryloxy, arylalkyloxy, oxyheterocyclic,
heterocyclic-substituted alkyloxy, thio C.sub.1-7 alkyl, thio
C.sub.3-10 cycloalkyl, thioaryl, thioheterocyclic, arylalkylthio,
hetero-cyclic-substituted alkylthio, formyl, hydroxylamino, cyano,
(thio)carboxylic acid or esters or thioesters or amides or
thioamides thereof; [0049] n is an integer from 0 to 6; and [0050]
R.sup.1 is a group selected from the group consisting of formyl,
acyl, thio-acyl, amide, thioamide, sulfonyl, sulfinyl, carboxylate,
thiocarboxylate, amino-substituted acyl, alkoxyalkyl, C.sub.3-10
cycloalkyl-alkyl, C.sub.3-10 cycloalkyl, dialkylaminoalkyl,
heterocyclic-substituted alkyl, acyl-substituted alkyl,
thioacylsubstituted alkyl, amido-substituted alkyl,
thioamido-substituted alkyl, carboxylato-substituted alkyl,
thiocarboxylato-substituted alkyl, (amino-substituted acyl)alkyl,
heterocyclic, carboxylic acid ester, .omega.-cyanoalkyl,
.omega.-carboxylic ester-alkyl, halo C.sub.1-7 alkyl, C.sub.2-7
alkenyl, C.sub.2-7 alkynyl, arylalkenyl, aryloxyalkyl, arylalkyl
and aryl, wherein the aryl moiety of each of said arylalkenyl,
aryloxyalkyl, arylalkyl and aryl radicals is optionally substituted
with one or more substituents independently selected from the group
consisting of halogen, C.sub.1-7 alkyl, C.sub.2-7alkenyl, C.sub.2-7
alkynyl, halo C.sub.1-7 alkyl, nitro, hydroxyl, sulfhydryl, amino,
C.sub.1-7 alkoxy, C.sub.3-10 cycloalkoxy, aryloxy, arylalkyloxy,
oxyheterocyclic, heterocyclic-substituted alkyloxy, thio C.sub.1-7
alkyl, thio C.sub.3-10 cycloalkyl, thioaryl, thioheterocyclic,
arylalkylthio, heterocyclic-substituted alkylthio, formyl,
carbamoyl, thiocarbamoyl, ureido, thioureido, sulfonamido,
hydroxylamino, alkoxyamino, mercapto-amino, thioalkylamino,
acylamino, thioacylamino, cyano, carboxylic acid or esters or
thioesters or halides or anhydrides or amides thereof,
thiocarboxylic acid or esters or thioesters or halides or
anhydrides or amides thereof, alkylamino, cycloalkylamino,
alkenylamino, cycloalkenyl-amino, alkynylamino, arylamino,
arylalkylamino, hydroxyalkylamino, mercaptoalkylamino, heterocyclic
amino, hydrazino, alkylhydrazino and phenylhydrazino.
[0051] For easiness of understanding the set of compounds of
general formula (I) may be sub-divided into six sub-sets of
compounds, depending upon (i) the nature of Y.sub.1 and Y.sub.2,
and (ii) when Y.sub.1 and Y.sub.2 are both oxygen, upon the number
and location of substituents on the pteridine ring. More
specifically, these six sub-sets of compounds of general formula
(I) are as follows: [0052] a) trisubstituted lumazines wherein
substitutions are in positions 1, 3 and 6 on the pteridine ring,
[0053] b) trisubstituted lumazines wherein substitutions are in
positions 1, 3 and 7 on the pteridine ring, [0054] c) mono- and
poly-substituted 2-thiolumazines, 4-thiolumazines and
2,4-dithiolumazines, [0055] d) tetrasubstituted lumazines, [0056]
e) trisubstituted lumazines wherein substitutions are in positions
3, 6 and 7 on the pteridine ring, and [0057] f) disubstituted
lumazines wherein substitutions are in positions 1 and 7 on the
pteridine ring.
[0058] It should be noted, however, that these sub-sets of novel
compounds have in common the structural features present in the
general formula (I). They also have a potential specific biological
activity profile and consequent usefulness in medicinal
chemistry.
[0059] In a second embodiment, the present invention relates to the
unexpected finding that at least one desirable biological property
such as, but not limited to, the ability to decrease the
proliferation of lymphocytes, or to decrease T-cell activation, or
to decrease B-cell or monocytes or macrophages activation, or to
inhibit TNF-.alpha. and IL-1 release, is a common feature which is
not only present in the group of novel compounds defined in the
general formula (I), but also in a group of polysubstituted
pteridine-2,4-diones (lumazines), as well as mono- and
poly-substituted 2-thiolumazines, 4-thiolumazines and
2,4-dithiolumazines which is broader than the said group of novel
compounds, specifically in a group of compounds represented by the
general formula (IV): ##STR5## wherein: [0060] each of Y.sub.1 and
Y.sub.2 is independently selected from sulfur and oxygen; [0061]
each of R'.sub.1 and R'.sub.2 is a radical independently selected
from the group consisting of hydrogen; C.sub.1-7 alkyl; C.sub.2-7
alkenyl; aryl; alkylaryl; .omega.-hydroxy C.sub.1-7 alkyl;
.omega.-epoxy C.sub.1-7 alkyl; .omega.-carboxy C.sub.1-7 alkyl
(wherein the carboxy group may be acid, ester, thioester, acid
halide or amide); .omega.-cyano C.sub.1-7 alkyl; arylalkyl;
arylalkenyl; heterocyclic-substituted alkyl;
heterocyclic-substituted alkenyl; groups having the formula --S--R
(i.e. wherein a sulfur atom is attached to the nitrogen atom of the
pteridine ring) wherein R is a monovalent group selected from the
group consisting of C.sub.1-7 alkyl, aryl and C.sub.3-10 cycloalkyl
and wherein the said monovalent group is optionally substituted
with one or more substituents selected from the group consisting of
amino, amino-acid, alkylamino, arylamino, cycloalkylamino,
carboxylic acid, carboxylic ester, sulfonic acid and phosphonic
acid; and optionally substituted heterocyclic radicals; [0062] at
most one of R'.sub.1, R'.sub.2, R'.sub.3 and R'.sub.4 is hydrogen
when one or more of Y.sub.1 and Y.sub.2 is sulfur; [0063] at most
one of R'.sub.1 and R'.sub.2 is hydrogen when both Y.sub.1 and
Y.sub.2 are oxygen; [0064] each of R'.sub.3 and R'.sub.4 is an atom
or radical independently selected from the group consisting of
hydrogen; halogen; C.sub.1-4 alkyl; C.sub.2-7 alkenyl; C.sub.2-7
alkynyl; halo C.sub.1-4 alkyl; C.sub.1-4 alkoxy; C.sub.3-10
cycloalkoxy; aryloxy; arylalkyloxy; oxyheterocyclic;
heterocyclic-substituted alkyloxy; thio C.sub.1-7alkyl; thio
C.sub.3-10 cycloalkyl; thioaryl; thioheterocyclic; arylalkylthio;
heterocyclic-substituted alkylthio; hydroxylamino; acetal; formyl;
cyano; carboxylic acid; carboxylic acid esters, thioesters and
amides; thiocarboxylic acid; thiocarboxylic acid esters, thioesters
and amides; amino; C.sub.2-7 alkylamino; cycloalkylamino;
alkenylamino; cycloalkenylamino; alkynylamino; arylamino;
arylalkylamino; hydroxyalkylamino; mercaptoalkylamino; heterocyclic
amino; heterocyclic-substituted alkylamino; oximino; alkyloximino;
hydrazino; alkylhydrazino; phenylhydrazino; cysteinyl acid, esters
or amides; aryl optionally substituted with one or more
substituents selected from the group consisting of halogen,
C.sub.1-7 alkyl, C.sub.2-7 alkenyl, C.sub.2-7 alkynyl, halo
C.sub.1-7 alkyl, C.sub.1-7 alkoxy, hydroxyl, sulfhydryl, amino,
C.sub.3-10 cycloalkoxy, aryloxy, arylalkyloxy, oxyheterocyclic,
heterocyclic-substituted alkyloxy, thio C.sub.1-7 alkyl, thio
C.sub.3-10 cycloalkyl, thioaryl, thioheterocyclic, arylalkylthio,
heterocyclic-substituted alkylthio, formyl, hydroxylamino, cyano,
carboxylic acid or esters or thioesters or amides thereof,
thiocarboxylic acid or esters or thioesters or amides thereof,
C.sub.1-7 alkylamino, cycloalkylamino, alkenylamino,
cycloalkenylamino, alkynylamino, arylamino, arylalkylamino,
hydroxyalkylamino, mercaptoalkylamino, heterocyclic amino,
hydrazino, alkylhydrazino and phenylhydrazino; optionally
substituted heterocyclic radicals; aryl or heterocyclic radicals
substituted with an aliphatic spacer between the pteridine ring and
the aryl or heterocyclic radical, whereby said aliphatic spacer
(linking group) is a branched or straight, saturated or unsaturated
aliphatic chain of 1 to 4 carbon atoms (e.g. a C.sub.1-4 alkylene)
which may contain one or more functions, atoms or radicals selected
from the group consisting of carbonyl (oxo), alcohol (hydroxyl),
ether, acetal, amino, imino, oximino, alkyloximino, amino-acid,
cyano, carboxylic acid or ester or thioester or amide, nitro, thio
C.sub.1-7 alkyl, thio C.sub.3-10 cycloalkyl, C.sub.1-7 alkylamino,
cycloalkylamino, alkenylamino, cycloalkenylamino, alkynylamino,
arylamino, arylalkylamino, hydroxyalkylamino, mercaptoalkylamino,
heterocyclic amino, hydrazino, alkylhydrazino, phenylhydrazino,
sulfonyl, sulfonamido and halogen; branched or straight, saturated
or unsaturated aliphatic chain of 1 to 7 carbon atoms containing
one or more atoms, functions or radicals selected from the group
consisting of carbonyl (oxo), alcohol (hydroxyl), ether, acetal,
amino, imino, oximino, alkyloximino, amino-acid, cyano, carboxylic
acid ester or amide, nitro, thio C.sub.1-7 alkyl, thio C.sub.3-10
cycloalkyl, C.sub.1-7 alkylamino, cycloalkylamino, alkenylamino,
cycloalkenylamino, alkynylamino, arylamino, arylalkylamino,
hydroxyalkylamino, mercaptoalkylamino, heterocyclic amino,
hydrazino, alkylhydrazino, phenylhydrazino, sulfonyl, sulfonamido
and halogen, and [0065] R'.sub.4 and R'.sub.3 may together form an
aryl radical being optionally substituted with one or more
substituents R.sub.a each independently selected from the group
consisting of amino, alkylamino, cycloalkylamino, alkenylamino,
cycloalkenylamino, alkynylamino, arylamino, arylalkylamino,
hydroxyalkylamino, mercaptoalkylamino, heterocyclic amino and
heterocyclic-substituted alkylamino, wherein each substituent
R.sub.a may further comprise one or more functions independently
selected from the group consisting of carbonyl, amino and carboxyl,
and wherein two adjacent substituents R.sub.a may together form an
heterocyclic radical.
[0066] Compounds of formula (IV) are highly active
immunosuppressive agents, antineoplastic agents or anti-viral
agents which, together with one or more pharmaceutically acceptable
carriers, may be formulated into pharmaceutical compositions for
the prevention or treatment of pathologic conditions such as, but
not limited to, immune and autoimmune disorders, organ and cells
transplant rejections, cell proliferative disorders, cardiovascular
disorders, disorders of the central nervous system and viral
diseases.
[0067] In a further embodiment, the present invention relates to
combined preparations containing at least one compound of formula
(IV) and one or more drugs such as immunosuppressant and/or
immunomodulator drugs, antineoplastic drugs, or antiviral agents.
In a further embodiment, the present invention relates to the
prevention or treatment of the above-cited pathologic conditions by
administering to the patient in need thereof an effective amount of
a compound of general formula (IV), optionally in the form of a
pharmaceutical composition or combined preparation with another
drug.
[0068] In a still further embodiment, the present invention relates
to processes and methods for making the novel poly-substituted
pteridine-2,4-diones (lumazines), as well as the novel mono- and
poly-substituted 2-thiolumazines, 4-thiolumazines and
2,4-dithiolumazines defined in general formula (I) and their
pharmaceutically acceptable salts and enantiomers.
BRIEF DESCRIPTION OF THE DRAWINGS
[0069] FIG. 1 represents a scheme for replacing a methyl
substituent R.sub.3 with an unsaturated partly aliphatic chain or
spacer in the 6-position of the pteridine ring of a substituted
lumazine.
[0070] FIG. 2 represents a scheme for replacing a carboxylic
substituent R.sub.3 in a substituted lumazine.
[0071] FIG. 3 represents a scheme for replacing a hydroxyl group
with an aryl substituent R.sub.4 in the 7-position of the pteridine
ring of a substituted lumazine.
[0072] FIG. 4 represents a scheme for performing various
alterations of a substituent R.sub.4 on a substituted lumazine
wherein the said R.sub.4 is either methyl or formyl.
[0073] FIG. 5 represents a scheme for performing various
alterations of a substituent R.sub.3 starting from a substituted
6-formyllumazine.
[0074] FIG. 6 represents a scheme for preparing a polysubstituted
lumazine wherein the substituent R.sub.4 is either alkylaryl or
hydroxyl and then performing various alterations thereon.
[0075] FIG. 7 represents a scheme for successively introducing
substituent R.sub.2 and R.sub.1 substituents into a
poly-substituted lumazine.
[0076] FIG. 8 represents a scheme for preparing
1-substituted-5,6-diaminouracil derivatives.
[0077] FIG. 9 represents a scheme for preparing a
1,3,6-trisubstituted lumazine with a halogen at position 7 of the
pteridine ring.
[0078] FIG. 10 represents a scheme for introducing a substituent
R.sub.2 or R.sub.1 onto a polysubstituted lumazine.
[0079] FIG. 11 represents a scheme for altering the side chain at
position 3 of the pteridine ring of a polysubstituted lumazine.
DEFINITIONS
[0080] As used herein, the terms "lumazine" and
"pteridine-2,4-dione" are interchangeable and designate any of the
tautomeric forms of 2,4-dioxopteridine or 2,4-dihydroxypteridine;
the term "2-thiolumazine" designate any of the tautomeric forms of
2-thioxo-4-oxo-pteridine; the term "4-thiolumazine" designate any
of the tautomeric forms of 2-oxo4-thioxo-pteridine; the term
"2,4-dithiolumazine" designate any of the tautomeric forms of
2,4-dithioxo-pteridine; the term "(thio)lumazine" is a generic
abbreviation for all sub-sets of compounds designated herein-above.
Unless otherwise stated herein, the term "polysubstituted" means
that two, three or four of the carbon atoms (thus respectively
"disubstituted", "trisubstituted or "tetrasubstituted") being in
any of positions 1, 3, 6 and 7 (according to standard atom
numbering for the pteridine ring) are substituted with an atom or
group other than hydrogen. The term "monosubstituted" means that
only one of the carbon atoms being in positions 1, 3, 6 or 7 is
substituted with an atom or group other than hydrogen.
[0081] As used herein with respect to a substituting group and
unless otherwise stated, the term "C.sub.1-7 alkyl" means straight
and branched chain saturated acyclic hydrocarbon monovalent
radicals or groups having from 1 to 7 carbon atoms such as, for
example, methyl, ethyl, propyl, n-butyl, 1-methylethyl (isopropyl),
2-methylpropyl (isobutyl), 1,1-dimethylethyl (ter-butyl),
2-methylbutyl, n-pentyl, dimethylpropyl, n-hexyl, 2-methylpentyl,
3-methylpentyl, n-heptyl and the like; optionally the carbon chain
length of such group may be extended to 20 carbon atoms; the term
"C.sub.1-4 alkyl" designate the corresponding radicals or groups
with only 1 to 4 carbon atoms, and so on.
[0082] As used herein with respect to a substituting radical, and
unless otherwise stated, the term "acyl" broadly refers to a
carbonyl (oxo) group adjacent to a C.sub.1-7 alkyl radical, a
C.sub.3-10 cycloalkyl radical, an aryl radical, an arylalkyl
radical or a heterocyclic radical, all of them being such as herein
defined; representative examples include acetyl, benzoyl, naphthoyl
and the like; similarly, the term "thioacyl" refers to a C.dbd.S
(thioxo) group adjacent to one of the said radicals.
[0083] As used herein with respect to a linking group and unless
otherwise stated, the term "C.sub.1-7 alkylene" means the divalent
hydrocarbon radical corresponding to the above defined C.sub.1-7
alkyl, such as methylene, bis(methylene), tris(methylene),
tetramethylene, hexamethylene and the like.
[0084] As used herein with respect to a substituting group and
unless otherwise stated, the term "C.sub.3-10 cycloalkyl" means a
mono- or polycyclic saturated hydrocarbon monovalent radical having
from 3 to 10 carbon atoms, such as for instance cyclopropyl,
cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl and
the like, or a C.sub.7-10 polycyclic saturated hydrocarbon
monovalent radical having from 7 to 10 carbon atoms such as, for
instance, norbornyl, fenchyl, trimethyltricycloheptyl or adamantyl;
the term "C.sub.5-8 cycloalkyl" designate the corresponding
radicals or groups with 5 to 8 carbon atoms, and so on.
[0085] As used herein with respect to a linking group and unless
otherwise stated, the term "C.sub.3-10 cycloalkylene" means the
divalent hydrocarbon radical corresponding to the above defined
C.sub.3-10 cycloalkyl, such as 1,2-cyclohexylene and
1,4-cyclohexylene.
[0086] As used herein with respect to a substituting group and
unless otherwise stated, the term "aryl" designate any mono- or
polyaromatic monovalent hydrocarbon radical having from 6 up to 30
carbon atoms such as but not limited to phenyl, naphthyl,
anthracenyl, phenantracyl, fluoranthenyl, chrysenyl, pyrenyl,
biphenylyl, terphenyl, picenyl, indenyl, indacenyl,
benzocyclobutenyl, benzocyclooctenyl and the like, including fused
benzo --C.sub.5-8 cycloalkyl radicals (the latter being as defined
above) such as, for instance, indanyl, tetrahydronaphtyl, fluorenyl
and the like, all of the said radicals being optionally substituted
with one or more substituents independently selected from the group
consisting of halogen, amino, halo C.sub.1-7 alkyl, hydroxyl,
sulfhydryl and nitro, such as for instance 4-fluorophenyl,
4-chlorophenyl, 3,4-dichlorophenyl, 2,6-diisopropyl-4-bromophenyl,
pentafluorophenyl, 4-trifluoromethylphenyl, 4-cyanophenyl,
2,6-dichlorophenyl, 2-fluorophenyl, 3-chlorophenyl,
3,5-dichlorophenyl and the like.
[0087] As used herein with respect to a linking group, and unless
otherwise stated, the term "arylene" means the divalent hydrocarbon
radical corresponding to the above defined aryl, such as phenylene,
naphtylene and the like.
[0088] As used herein with respect to a substituting radical
(including a combination of substituents in positions 6 and 7 of
the pteridine ring together with the carbon atoms in positions 6
and 7 of the pteridine ring), and unless otherwise stated, the term
"heterocyclic" means a mono- or polycyclic, saturated or
mono-unsaturated or polyunsaturated monovalent hydrocarbon radical
having from 2 up to 15 carbon atoms and including one or more
heteroatoms in one or more heterocyclic rings, each of said rings
having from 3 to 10 atoms (and optionally further including one or
more heteroatoms attached to one or more carbon atoms of said ring,
for instance in the form of a carbonyl or thiocarbonyl or
selenocarbonyl group and/or to one or more heteroatoms of said
ring, for instance in the form of a sulfone, sulfoxide, N-oxide,
phosphate, phosphonate or selenium oxide group), each of said
heteroatoms being independently selected from the group consisting
of nitrogen, oxygen, sulfur, selenium and phosphorus, also
including radicals wherein a heterocyclic ring is fused to one or
more aromatic hydrocarbon rings for instance in the form of
benzo-fused, dibenzo-fused and naphto-fused heterocyclic radicals;
within this definition are included heterocyclic radicals such as,
but not limited to, diazepinyl, oxadiazinyl, thiadiazinyl,
dithiazinyl, triazolonyl, diazepinonyl, triazepinyl, triazepinonyl,
tetrazepinonyl, benzoquinolinyl, benzothiazinyl, benzothiazinonyl,
benzoxathiinyl, benzodioxinyl, benzodithiinyl, benzoxazepinyl,
benzothiazepinyl, benzo-diazepinyl, benzodioxepinyl,
benzodithiepinyl, benzoxazocinyl, benzothiazocinyl,
benzodiazocinyl, benzoxathiocinyl, benzodioxocinyl,
benzotrioxepinyl, benzoxathiazepinyl, benzoxadiazepinyl,
benzothiadiazepinyl, benzotriazepinyl, benzoxathiepinyl,
benzotriazinonyl, benzoxazolinonyl, azetidinonyl, azaspiro-undecyl,
dithiaspirodecyl, selenazinyl, selenazolyl, selenophenyl,
hypoxanthinyl, azahypoxanthinyl, bipyrazinyl, bipyridinyl,
oxazolidinyl, diselenopyrimidinyl, benzopyrenyl, benzopyranonyl,
benzophenazinyl, benzoquinolizinyl, dibenzocarbazolyl,
dibenzoacridinyl, dibenzophenazinyl, dibenzothiepinyl,
dibenzo-oxepinyl, dibenzopyranonyl, dibenzoquinoxalinyl,
dibenzothiazepinyl, dibenzoisoquinolinyl, tetraazaadamantyl,
thiatetraazaadamantyl, oxauracil, oxazinyl, dibenzothiophenyl,
dibenzofuranyl, oxazolinyl, oxazolonyl, azaindolyl, azolonyl,
thiazolinyl, thiazolonyl, thiazolidinyl, thiazanyl, pyrimidonyl,
thiopyrimidonyl, thiamorpholinyl, azlactonyl, naphtindazolyl,
naphtindolyl, naphtothiazolyl, naphtothioxolyl, naphtoxindolyl,
naphtotriazolyl, naphtopyranyl, indolinyl, indolizidinyl,
oxabicycloheptyl, azabenzimidazolyl, azacycloheptyl, azacyclooctyl,
azacyclononyl, azabicyclononyl, tetrahydrofuryl, tetrahydropyranyl,
tetrahydropyronyl, tetrahydroquinoleinyl, tetrahydrothienyl and
dioxide thereof, dihydrothienyl dioxide, dioxindolyl, dioxinyl,
dioxenyl, dioxazinyl, thioxanyl, thioxolyl, thio-urazolyl,
thiotriazolyl, thiopyranyl, thiopyronyl, coumarinyl, quinoleinyl,
oxyquinoleinyl, quinuclidinyl, xanthinyl, dihydropyranyl,
benzodihydro-furyl, benzothiopyronyl, benzothiopyranyl,
benzoxazinyl, benzoxazolyl, benzodioxolyl, benzodioxanyl,
benzothiadiazolyl, benzotriazinyl, benzothiazolyl, benzoxazolyl,
phenothioxinyl, phenothiazolyl, phenothienyl, phenopyronyl,
phenoxazolyl, pyridinyl, dihydropyridinyl, tetrahydropyridinyl,
piperidinyl, morpholinyl, thiomorpholinyl, pyrazinyl, pyrimidinyl,
pyridazinyl, triazinyl, tetrazinyl, triazolyl, benzotriazolyl,
tetrazolyl, imidazolyl, pyrazolyl, thiazolyl, thiadiazolyl,
isothiazolyl, oxazolyl, oxadiazolyl, pyrrolyl, furyl, dihydrofuryl,
furoyl, hydantoinyl, dioxolanyl, dioxolyl, dithianyl, dithienyl,
dithiinyl, thienyl, indolyl, indazolyl, benzofuryl, quinolyl,
quinazolinyl, quinoxalinyl, carbazolyl, phenoxazinyl,
phenothiazinyl, xanthenyl, purinyl, benzothienyl, naphtothienyl,
thianthrenyl, pyranyl, pyronyl, benzopyronyl, isobenzofuranyl,
chromenyl, phenoxathiinyl, indolizinyl, quinolizinyl, isoquinolyl,
phthalazinyl, naphthiridinyl, cinnolinyl, pteridinyl, carbolinyl,
acridinyl, perimidinyl, phenanthrolinyl, phenazinyl,
phenothiazinyl, imidazolinyl, imidazolidinyl, benzimidazolyl,
pyrazolinyl, pyrazolidinyl, pyrrolinyl, pyrrolidinyl, piperazinyl,
uridinyl, thymidinyl, cytidinyl, azirinyl, aziridinyl, diazirinyl,
diaziridinyl, oxiranyl, oxaziridinyl, dioxiranyl, thiiranyl,
azetyl, dihydroazetyl, azetidinyl, oxetyl, oxetanyl, thietyl,
thietanyl, oxetanonyl, diazabicyclooctyl, diazetyl, diaziridinonyl,
diaziridinethionyl, chromanyl, chroma-nonyl, thiochromanyl,
thiochromanonyl, thiochromenyl, benzofuranyl, benzisothiazolyl,
benzocarbazolyl, benzochromonyl, benzisoalloxazinyl,
benzocoumarinyl, thiocoumarinyl, phenometoxazinyl,
phenoparoxazinyl, phentriazinyl, thiodiazinyl, thiodiazolyl,
indoxyl, thioindoxyl, benzodiazinyl (e.g. phtalazinyl), phtalidyl,
phtalimidinyl, phtalazonyl, alloxazinyl, dibenzopyronyl (i.e.
xanthonyl), xanthionyl, isatyl, isopyrazolyl, isopyrazolonyl,
urazolyl, urazinyl, uretinyl, uretidinyl, succinyl, succinimido,
benzylsultimyl, and benzylsultamyl and the like, including all
possible isomeric forms thereof, wherein each carbon atom of the
said heterocyclic ring may be independently substituted with a
substituent selected from the group consisting of halogen, nitro,
C.sub.1-7 alkyl (optionally containing one or more functions or
radicals selected from the group consisting of carbonyl (oxo),
alcohol (hydroxyl), ether (alkoxy), acetal, amino, imino, oximino,
alkyloximino, amino-acid, cyano, carboxylic acid ester or amide,
nitro, thio C.sub.1-7 alkyl, thio C.sub.3-10 cycloalkyl, C.sub.1-7
alkylamino, cycloalkylamino, alkenylamino, cycloalkenylamino,
alkynylamino, arylamino, arylalkylamino, hydroxylalkylamino,
mercaptoalkylamino, heterocyclic amino, hydrazino, alkylhydrazino,
phenyl-hydrazino, sulfonyl, sulfonamido and halogen), C.sub.3-7
alkenyl, C.sub.2-7 alkynyl, halo C.sub.1-7 alkyl, C.sub.3-10
cycloalkyl, aryl, arylalkyl, alkylaryl, alkylacyl, arylacyl,
hydroxyl, amino, C.sub.1-7 alkylamino, cycloalkylamino,
alkenylamino, cycloalkenylamino, alkynylamino, arylamino,
arylalkylamino, hydroxyalkylamino, mercaptoalkylamino, heterocyclic
amino, hydrazino, alkylhydrazino, phenylhydrazino, sulfhydryl,
C.sub.1-7 alkoxy, C.sub.3-10 cycloalkoxy, aryloxy, arylalkyloxy,
oxyheterocyclic, heterocyclic-substituted alkyloxy, thio C.sub.1-7
alkyl, thio C.sub.3-10 cycloalkyl, thioaryl, thioheterocyclic,
arylalkylthio, heterocyclic-substituted alkylthio, formyl,
hydroxylamino, cyano, carboxylic acid or esters or thioesters or
amides thereof, thiocarboxylic acid or esters or thioesters or
amides thereof; depending upon the number of unsaturations in the
heterocyclic ring having 3 to 10 atoms, heterocyclic radicals may
be sub-divided into heteroaromatic (or "heteroaryl") radicals and
non-aromatic heterocyclic radicals; when a heteroatom of the said
non-aromatic heterocyclic radical is nitrogen, the latter may be
substituted with a substituent selected from the group consisting
of C.sub.1-7 alkyl, C.sub.3-10 cycloalkyl, aryl, arylalkyl,
alkylaryl, alkylacyl and arylacyl.
[0089] As used herein with respect to a substituting group and
unless otherwise stated, the terms "C.sub.1-7 alkoxy", "C.sub.2-7
alkenyloxy", "C.sub.2-7 alkynyloxy", "C.sub.3-10 cycloalkoxy",
"aryloxy", "arylalkyloxy", "oxyheterocyclic", "thio C.sub.1-7
alkyl", "thio C.sub.3-10 cycloalkyl", "arylthio", "arylalkylthio"
and "thioheterocyclic" refer to substituents wherein a C.sub.1-7
alkyl, C.sub.2-7 alkenyl or C.sub.2-7 alkynyl (optionally the
carbon chain length of such group may be extended to 20 carbon
atoms), respectively a C.sub.3-10 cycloalkyl, aryl, arylalkyl or
heterocyclic radical (each of them such as defined herein), are
attached to an oxygen atom or a divalent sulfur atom through a
single bond, such as but not limited to methoxy, ethoxy, propoxy,
butoxy, pentoxy, isopropoxy, sec-butoxy, tert-butoxy, isopentoxy,
cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, thiomethyl,
thioethyl, thiopropyl, thiobutyl, thiopentyl, thiocyclopropyl,
thiocyclobutyl, thiocyclopentyl, thiophenyl, cresoxy, phenyloxy,
benzyloxy, mercaptobenzyl and the like.
[0090] As used herein with respect to a substituting atom and
unless otherwise stated, the term halogen means any atom selected
from the group consisting of fluorine, chlorine, bromine and
iodine.
[0091] As used herein with respect to a substituting group and
unless otherwise stated, the term "halo C.sub.1-7 alkyl" means a
C.sub.1-7 alkyl radical (such as above defined, i.e. optionally the
carbon chain length of such group may be extended to 20 carbon
atoms) in which one or more hydrogen atoms are independently
replaced by one or more halogens (preferably fluorine, chlorine or
bromine), such as but not limited to difluoromethyl,
trifluoromethyl, trifluoroethyl, octafluoropentyl,
dodecafluoroheptyl, dichloromethyl and the like; the term "halo
C.sub.1-4 alkyl" designate the corresponding radical with only 1 to
4 carbon atoms, and so on.
[0092] As used herein with respect to a substituting group and
unless otherwise stated, the terms "C.sub.2-7 alkenyl" and
"aliphatic unsaturated hydrocarbon radical with 2 to 7 carbon
atoms" are interchangeable and designate a straight or branched
acyclic hydrocarbon monovalent radical having one or more
ethylenical unsaturations and having from 2 to 7 carbon atoms such
as, for example, vinyl, 2-propenyl, 3-butenyl, 2-butenyl,
2-pentenyl, 3-pentenyl, 3-methyl-2-butenyl, 3-hexenyl, 2-hexenyl,
2-heptenyl, butadienyl, pentadienyl, hexadienyl, heptadienyl,
heptatrienyl and the like, including all possible isomers thereof;
optionally the carbon chain length of such group may be extended to
20 carbon atoms; the term "C.sub.3-7 alkenyl" designate the
corresponding radical with only 3 to 7 carbon atoms, and so on.
[0093] As used herein with respect to a substituting group and
unless otherwise stated, the terms "C.sub.3-10 cycloalkenyl" means
a monocyclic mono- or polyunsaturated hydrocarbon monovalent
radical having from 3 to 8 carbon atoms, such as for instance
cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclopentadienyl,
cyclohexenyl, cyclohexadienyl, cycloheptenyl, cycloheptadienyl,
cycloheptatrienyl, cyclooctenyl, cyclooctadienyl, cyclooctatrienyl,
1,3,5,7-cyclooctatetraenyl and the like, or a C.sub.7-10 polycyclic
mono- or polyunsaturated hydrocarbon monovalent radical having from
7 to 10 carbon atoms such as dicyclopentadienyl, fenchenyl
(including all isomers thereof, such as .alpha.-pinolenyl),
bicyclo[2.2.1]hept-2-enyl, bicyclo[2.2.1]hepta-2,5-dienyl,
cyclofenchenyl and the like.
[0094] As used herein with respect to a substituting group and
unless otherwise stated, the term "C.sub.2-7 alkynyl" defines
straight and branched chain hydrocarbon radicals containing one or
more triple bonds and having from 2 to 7 carbon atoms such as, for
example, acetylenyl, 2-propynyl, 3-butynyl, 2-butynyl, 2-pentynyl,
3-pentynyl, 3-methyl-2-butynyl, 3-hexynyl, 2-hexynyl and the like
and all possible isomers thereof; optionally the carbon chain
length of such group may be extended to 20 carbon atoms.
[0095] As used herein with respect to a substituting group and
unless otherwise stated, the terms "arylalkyl", "arylalkenyl",
"heterocyclic-substituted alkyl" and "heterocyclic-substituted
alkenyl" refer to an aliphatic saturated or ethylenically
unsaturated hydrocarbon monovalent radical (preferably a C.sub.1-7
alkyl or C.sub.3-10 cycloalkyl or C.sub.2-7 alkenyl or C.sub.3-10
cycloalkenyl such as defined above, i.e. optionally the carbon
chain length of such group may be extended to 20 carbon atoms),
onto which an aryl radical or respectively a heterocyclic radical
(such as defined above) is already bonded, and wherein the said
aliphatic radical and/or the said aryl or heterocyclic radical may
be optionally substituted with one or more substituents
independently selected from the group consisting of C.sub.1-4
alkyl, trifluoromethyl, halogen, amino, nitro, hydroxyl, sulfhydryl
and nitro, such as but not limited to benzyl, 4-chlorobenzyl,
2-fluorobenzyl, 4-fluorobenzyl, 3,4-dichlorobenzyl,
2,6-dichlorobenzyl, 4-ter-butylbenzyl, 3-methylbenzyl,
4-methylbenzyl, phenylpropyl, 1-naphtylmethyl, phenylethyl,
1-amino-2-phenylethyl, 1-amino-2-[4-hydroxy-phenyl]ethyl,
1-amino-2-[indol-2-yl]ethyl, styryl, pyridylmethyl (including all
isomers thereof), pyridylethyl, 2-(2-pyridyl)isopropyl,
oxazolylbutyl, 2-thienylmethyl, pyrrolylethyl, morpholinyl-ethyl,
imidazol-1-yl-ethyl, benzodioxolylmethyl and 2-furylmethyl.
[0096] As used herein with respect to a substituting group and
unless otherwise stated, the terms "alkylcycloalkyl",
"alkenyl(hetero)aryl", "alkyl(hetero)aryl", and "alkyl-substituted
heterocyclic" refer respectively to an aryl, heteroaryl, cycloalkyl
or heterocyclic radical (such as defined above) onto which are
already bonded one or more aliphatic saturated or unsaturated
hydrocarbon monovalent radicals, preferably one or more C.sub.1-7
alkyl, C.sub.2-7 alkenyl or C.sub.3-10 cycloalkyl radicals as
defined above, such as, but not limited to, o-toluyl, m-toluyl,
p-toluyl, 2,3-xylyl, 2,4-xylyl, 3,4-xylyl, o-cumenyl, m-cumenyl,
p-cumenyl, o-cymenyl, m-cymenyl, p-cymenyl, mesityl,
ter-butylphenyl, lutidinyl (i.e. dimethylpyridyl),
2-methylaziridinyl, methylbenzimidazolyl, methylbenzofuranyl,
methylbenzothiazolyl, methylbenzotriazolyl, methylbenzoxazolyl,
methylcyclohexyl and menthyl.
[0097] As used herein with respect to a substituting radical, and
unless otherwise stated, the term "alkoxyaryl" refers to an aryl
radical (such as defined above) onto which is (are) bonded one or
more C.sub.1-7 alkoxy radicals as defined above, preferably one or
more methoxy radicals, such as, but not limited to,
2-methoxyphenyl, 3-methoxyphenyl, 4-methoxyphenyl,
3,4-dimethoxyphenyl, 2,4,6-trimethoxyphenyl, methoxynaphtyl and the
like.
[0098] As used herein with respect to a substituting group and
unless otherwise stated, the terms "alkylamino", "cycloalkylamino",
"alkenylamino", "cycloalkenylamino", "arylamino", "arylalkylamino",
"heterocyclic amino", "hydroxyalkylamino", "mercaptoalkylamino" and
"alkynylamino" mean that one or even two C.sub.1-7 alkyl,
C.sub.3-10 cycloalkyl, C.sub.2-7 alkenyl, C.sub.3-10 cycloalkenyl,
aryl, arylalkyl, heterocyclic, hydroxy C.sub.1-7 alkyl, mercapto
C.sub.1-7 alkyl or C.sub.2-7 alkynyl radicals (each of them as
defined herein, respectively) are attached to a nitrogen atom
through a single bond or, in the case of heterocyclic, include a
nitrogen atom, such as but not limited to, anilino, benzylamino,
methylamino, dimethylamino, ethylamino, isopropylamino,
propenylamino, n-butylamino, ter-butylamino, dibutylamino,
morpholinoalkylamino, morpholinyl, piperidinyl, piperazinyl,
hydroxymethylamino and ethynylamino; this definition also includes
mixed amino radicals wherein the nitrogen atom is attached to two
such radicals belonging to two different sub-set of radicals, e.g.
an alkyl radical and an alkenyl radical.
[0099] As used herein with respect to a substituting group and
unless otherwise stated, the terms "(thio)carboxylic acid ester",
"(thio)carboxylic acid thioester" and "(thio)carboxylic acid amide"
refer to radicals wherein the carboxyl or thiocarboxyl group is
directly attached to the pteridine ring (e.g. in the 6- and/or
7-position) and wherein said carboxyl or thiocarboxyl group is
bonded to the hydrocarbonyl residue of an alcohol, a thiol, a
polyol, a phenol, a thiophenol, a primary or secondary amine, a
polyamine, an amino-alcohol or ammonia, the said hydrocarbonyl
residue being selected from the group consisting of alkyl, alkenyl,
alkynyl, cycloalkyl, cycloalkenyl, aryl, arylalkyl, alkylaryl,
alkylamino, cycloalkylamino, alkenylamino, cycloalkenylamino,
arylamino, arylalkylamino, heterocyclic amino, hydroxyalkylamino,
mercaptoalkylamino or alkynylamino (such as above defined,
respectively).
[0100] As used herein with respect to a substituting group and
unless otherwise stated, the term "amino-acid" refers to a radical
derived from a molecule having the chemical formula
H.sub.2N--CHR--COOH, wherein R is the side group of atoms
characterizing the amino-acid type; said molecule may be one of the
20 naturally-occurring amino-acids or any non naturally-occurring
amino-acid.
[0101] As used herein and unless otherwise stated, the term
"enantiomer" means each individual optically active form of a
compound of the invention, having an optical purity or enantiomeric
excess (as determined by methods standard in the art) of at least
80% (i.e. at least 90% of one enantiomer and at most 10% of the
other enantiomer), preferably at least 90% and more preferably at
least 98%.
DETAILED DESCRIPTION OF THE INVENTION
[0102] In one aspect, the present invention requires a
3-alkyl-6-aminouracil or a 3-alkenyl-6-uracil as a starting
material. Since the procedure of WO 03/067257 achieves
3-methyl-6-aminouracil only with a poor yield, one aspect of the
invention comprises a method of making a 3-alkyl-6-aminouracil or a
3-alkenyl-6-uracil in high yield by reacting, in the presence of a
catalytic amount of ammonium sulfate, 6-aminouracil with a molar
excess of an alkyl halide or alkenyl halide (wherein the halide is
preferably iodine, and the alkyl or alkenyl group may have from 1
to 7 carbon atoms) at reflux temperature in the presence of
1,1,1,3,3,3-hexamethyldisilazane as a solvent, and then separating
the precipitate formed after quenching the reaction e.g. with
sodium bicarbonate.
[0103] In the first embodiment of the invention, the novel
poly-substituted pteridine-2,4-diones (lumazines), as well as the
novel mono- and poly-substituted 2-thiolumazines, 4-thiolumazines
and 2,4-dithiolumazines are as defined in the general formula (I),
wherein each of the substituents Y.sub.1, Y.sub.2, R.sub.1,
R.sub.2, R.sub.3 and R.sub.4, and optionally any of n, R.sub.0 and
R.sup.1, may correspond to any of the definitions given herein, in
particular with any of the individual meanings (such as illustrated
above) of generic terms such as but not limited to "C.sub.1-7
alkyl", "C.sub.2-7 alkenyl", "C.sub.2-7 alkynyl", "aryl",
"heterocyclic", "alkylaryl", "arylalkyl", "alkylamino",
"cycloalkylamino", "alkenylamino", "alkynylamino", "arylamino",
"aryl-alkylamino", C.sub.1-7 alkoxy", "C.sub.3-10 cycloalkoxy",
"thio C.sub.1-7 alkyl", "thio C.sub.3-10 cycloalkyl", "halo
C.sub.1-7 alkyl", "amino-acid" and the like.
[0104] When a mixture of enantiomers of the pteridinediones
(lumazines), 2-thiolumazines, 4-thiolumazines or
2,4-dithiolumazines having the general formula (I) according to the
invention is obtained during their synthesis, the said mixture may
be separated by means and methods standard in the art, e.g. liquid
chromatography using one or more suitable chiral stationary phases.
The latter include, for example, polysaccharides, in particular
cellulose or amylose derivatives. Commercially available
polysaccharide based chiral stationary phases are ChiralCel.TM. CA,
OA, OB, OC, OD, OF, OG, OJ and OK, and Chiralpak.TM. AD, AS, OP(+)
and OT(+). Appropriate eluents or mobile phases for use in
combination with said polysaccharide chiral stationary phases are
hydrocarbons such as hexane and the like, optionally admixed with
an alcohol such as ethanol, isopropanol and the like. The above
mixture of enantiomers may alternatively be separated by forming
diastereoisomers, followed by separation of the diastereoisomers,
e.g. by differential crystallization or chromatography. The
resolving agent may be cleaved from the separated diastereoisomers,
e.g. by treatment with acids or bases, to generate the pure
enantiomers of the compounds of the invention.
[0105] Some preferred polysubstituted pteridine-2,4-diones
(lumazines), as well as mono- and polysubstituted 2-thiolumazines,
4-thiolumazines and 2,4-dithiolumazines having the general formula
(I) according to the invention are more specifically illustrated in
the following examples and defined in the following claims. For
instance, useful species include those wherein: [0106] R.sub.1 and
R.sub.2 are independently benzyl, phenyl, 2-phenylethyl, butyric
acid or ester, butyronitrile, 2-hydroxyethyl, ethyl,methyl, and/or
[0107] R.sub.4 is chloro, hydroxyl, phenoxy, and/or [0108] R.sub.3
is substituted phenyl.
[0109] The present invention further provides processes and methods
for making the novel polysubstituted pteridine-2,4-diones
(lumazines), as well as the novel mono- and polysubstituted
2-thiolumazines, 4-thiolumazines and 2,4-dithiolumazines having the
general formula (I). As a general rule, the preparation of these
(thio)lumazines is based on the principle that, starting from a
lumazine or (di)thiolumazine or a lumazine or (di)thiolumazine
precursor, e.g. an adequately substituted uracil or thiouracil,
each of the substituents R.sub.1, R.sub.2, R.sub.3 and R.sub.4 may
be introduced separately without adversely influencing the presence
of a substituent already introduced or the capacity to introduce
further substituents later on. Therefore a process for making a
(thio)lumazine having the general formula (I) usually consists of
one (e.g. in the case of mono-substituted thiolumazines) or more
(in the case of polysubstituted lumazines and thiolumazines)
reaction steps for successively introducing one or more of the
substituents R.sub.1, R.sub.2, R.sub.3 and R.sub.4 into a compound
selected from the group consisting of lumazine, 2-thiolumazine,
4-thiolumazine, 2,4-dithiolumazine and known substituted
(thio)lumazines, or starting from suitable (thio)lumazine
precursors (such as uracils and thiouracils) already bearing the
desired substituents, each reaction step being optionally, if
needed, sub-divided into one or more sub-steps involving
intermediate substituted (thio)lumazines.
[0110] A limited number of methods are already known in the art for
introducing a substituent R.sub.1, R.sub.2, R.sub.3 or R.sub.4 to
form a (substituted) lumazine or a substituted (thio)lumazine
precursor, and a still more limited number of methods are known in
the art for introducing a substituent R.sub.1, R.sub.2, R.sub.3 or
R.sub.4 into a 2-thiolumazine, 4-thiolumazine or
2,4-dithiolumazine, all of these methods being disclosed in the
prior documents referred to in the background of the invention.
These methods may be applied successfully to the preparation of
compounds having the general formula (I). Other methods have been
developed by the present inventors, which may be used alternatively
or may be combined with the former methods (depending upon the
targeted final compound) and will now be explained by reference to
the appended FIGS. 1 to 7 relating to substituted lumazines
wherein, unless otherwise stated hereinafter, R.sub.1, R.sub.2,
R.sub.3 or R.sub.4 are as defined in the summary of the invention.
The same methods may be applied starting from the few substituted
thiolumazines or dithiolumazines which are already known in the
art. In the description of the reaction steps involved in each
figure, reference may be made to the use of certain catalysts
and/or certain types of solvents. It should be understood that each
catalyst mentioned should be used in a catalytic amount well known
to the skilled person with respect to the type of reaction
involved. Solvents that may be used in the following reaction steps
include various kinds of organic solvents such as protic solvents,
polar aprotic solvents and non-polar solvents as well as aqueous
solvents which are inert under the relevant reaction conditions.
More specific examples include aromatic hydrocarbons, chlorinated
hydrocarbons, ethers, aliphatic hydrocarbons, alcohols, esters,
ketones, amides, water or mixtures thereof, as well as
supercritical solvents such as carbon dioxide (while performing the
reaction under supercritical conditions). The suitable reaction
temperature and pressure conditions applicable to each kind of
reaction step will not be detailed herein but do not depart from
the relevant conditions already known to the skilled person with
respect to the type of reaction involved.
[0111] FIG. 1 represents a scheme for (i) replacing a methyl
substituent R.sub.3 with an unsaturated partly aliphatic chain or
spacer in the 6-position of the pteridine ring and optionally
further (ii) inserting an oxygen atom between the R.sub.4
substituent and the pteridine ring in the 7-position thereof.
The R.sub.3 substituent replacement occurs via a succession of
reaction steps as follows. In reaction step (a), a 6-methyl
substituted lumazine is reacted:
[0112] either with an halogen, preferably chlorine or bromine, in
the presence of a protic solvent, or [0113] with an
N-halosuccinimide, wherein the halogen is preferably chlorine or
bromine, in the presence of a protic or aprotic solvent. Then in
reaction step (b), the reaction product of step (a) is reacted, in
the presence of a non-polar solvent, preferably toluene, xylene or
nitro-methane, with a phosphine selected from the group consisting
of trialkylphosphines (alkyl.sub.3P), triarylphosphines
(aryl.sub.3P), tricycloalkylphosphines (cycloalkyl.sub.3P) and
trialkylphosphites (alkoxy.sub.3P), or the corresponding arsines,
thus resulting in an intermediate compound wherein R.sub.5 is
selected from the group consisting of (aryl).sub.3 P.sup.+X.sup.-,
(alkyl).sub.3 P.sup.+X.sup.-, (cycloalkyl).sub.3 P.sup.+X.sup.-,
(alkyloxy).sub.2PO, (aryl).sub.3 As.sup.+X.sup.-, (alkyl).sub.3
As.sup.+X.sup.- and (cycloalkyl).sub.3 As.sup.+X.sup.-.
[0114] Then in the combined reaction steps (c) and (d), this
intermediate compound is reacted, in the presence of a catalyst,
with alkyl- or aryl- or alkylaryl- or heterocyclic- or
alkoxycarbonyl-aldehyde or ketone. Suitable 10 aliphatic, aromatic
or heteroaromatic aldehydes and ketones include (in the following
non-exhaustive list, use of the plural is meant to include all
possible isomers) benzaldehyde, mono- and polyhalogenated
benzaldehydes, cyanobenzaldehydes, substituted or non-substituted
amino-benzaldehydes, mono- and dinitrobenzaldehydes, mono- and
polyalkoxybenzaldehydes, mono- and polyalkylated benzaldehydes,
carboxylated (esters and amides) benzaldehydes,
aryloxybenzaldehydes, 2-fluorene-carboxaldehyde, naphthaldehydes,
alkoxynaphthaldehydes, N-ethyl-3-carbazole-carboxaldehyde,
4-formylcinnamic acid, alkylthiobenzaldehydes,
2-formylbenzenesulfonic acid, methylformyl-benzoate,
acetaminobenzaldehyde, aryloxyalkylbenzaldehydes,
acetamido-benzaldehyde, alkylsulfonylbenzaldehyde, propionaldehyde,
butyraldehyde and the corresponding ketones. The catalyst used in
the combined reaction steps (c) and (d) may be selected from the
group consisting of alkoxy-alkaline metals (e.g. wherein the metal
is Li, Na or K), DBU (1,8-diazabicyclo[5.4.0]undec-7-ene), DBN
(1,5-diazabicyclo[4.3.0]non-5-ene), guanidine and strong bases such
as butyllithium. The combined reaction steps (c) and (d) result in
a compound wherein R.sub.6 is selected from the group consisting of
alkyl, aryl, alkylaryl, heterocyclic and alkoxycarbonyl and
wherein, depending on whether an aldehyde or a ketone was reacted,
R.sub.7 is selected from the group consisting of hydrogen, alkyl
and aryl.
[0115] Then in reaction step (e), this compound is reacted with an
halogen, preferably chlorine or bromine, in the presence of a
chlorinated solvent such as carbon tetrachloride or chloroform.
Finally in reaction step (f), the replacement of the R.sub.4
substituent is performed by reacting the product from step (e) with
an alkoxy-alkaline metal (e.g. wherein the metal is Li, Na or K),
thus resulting in a compound wherein R.sub.8 is alkyl.
[0116] FIG. 2 represents a scheme for replacing a carboxylic
substituent R.sub.3 in a substituted lumazine with a group having
the formula COCHZ.sub.1Z.sub.2, wherein each of Z.sub.1 and Z.sub.2
may be independently selected from the group consisting of
alkylcarbonyl, arylcarbonyl, alkoxycarbonyl, aryloxycarbonyl and
cyano. In a first reaction step (a), a 6-carboxylic
acid-substituted lumazine is reacted with: [0117] either a compound
selected from the group consisting of SOX.sub.2, (COX).sub.2,
PX.sub.3 and PX.sub.5 (preferably wherein X is Cl or Br) in the
presence of a solvent, [0118] or triphenylphosphine in the presence
of carbon tetrachloride. In a second reaction step (b), the
halocarbonyl product from step (a) is reacted with a methylene
Z.sub.1Z.sub.2 ethoxy-magnesium salt or alkaline metal salt
(preferably wherein the metal is Na, K or Li) in the presence of an
aprotic solvent.
[0119] FIG. 3 represents a scheme for replacing a hydroxyl group
with an aryl substituent R.sub.4 in the 7-position of the pteridine
ring of a substituted lumazine. In a first reaction step (a), a
7-hydroxyl, 6-carboxyaryl substituted lumazine is reacted with an
aromatic hydrocarbon in the presence of a catalyst such as aluminum
trichloride and a chlorinated solvent, then in a second step (b)
the product of step (a) is reacted with KMnO.sub.4 in the presence
of a solvent such as dioxane.
[0120] FIG. 4 represents a scheme for performing various
alterations of a substituent R.sub.4 starting from a known
substituted lumazine wherein the said R.sub.4 is either methyl or
formyl. In reaction step (a), a 7-methyl substituted lumazine is
reacted: [0121] either with an halogen, preferably chlorine or
bromine, in the presence of a protic solvent, or [0122] with an
N-halosuccinimide, wherein the halogen is preferably chlorine or
bromine, in the presence of a protic or aprotic solvent. Then in
reaction step (b), the reaction product of step (a) is reacted, in
the presence of a non-polar solvent, preferably toluene, xylene or
nitromethane, with a phosphine selected from the group consisting
of trialkylphosphines (alkyl.sub.3P), triarylphosphines
(aryl.sub.3P), tricycloalkylphosphines (cycloalkyl.sub.3P) and
trialkoxyphosphines (alkoxy.sub.3P), or the corresponding arsines,
thus resulting in an intermediate compound wherein R.sub.5 is
selected from the group consisting of (aryl).sub.3 P.sup.+X.sup.-,
(alkyl).sub.3 P.sup.+X.sup.-, (cycloalkyl).sub.3 P.sup.+X.sup.-,
(alkyloxy).sub.2PO, (aryl).sub.3 As.sup.+X.sup.-, (alkyl).sub.3
As.sup.+X.sup.- and (cycloalkyl).sub.3 As.sup.+X.sup.-.
[0123] Then in the combined reaction steps (c) and (d), this
intermediate compound is reacted, in the presence of a catalyst,
with an alkyl- or aryl- or alkylaryl- or heterocyclic- or
alkoxycarbonyl-aldehyde or ketone which, alike the catalyst, may be
as defined herein-above with reference to the description of FIG.
1. The combined reaction steps (c) and (d) result in a compound
wherein R.sub.6 is selected from the group consisting of alkyl,
aryl, alkylaryl, heterocyclic and alkoxycarbonyl and wherein,
depending on whether an aldehyde or a ketone was reacted, R.sub.7
is selected from the group consisting of hydrogen, alkyl and aryl.
This same compound may also be prepared from a 7-formyl substituted
lumazine through the combined reaction steps (g) and (h), using
first an alkyl-, alkylaryl-, alkylheterocyclic-,
alkoxycarbonylalkyl-, aryloxycarbonylalkyl-R.sub.9 wherein R.sub.9
is selected from the group consisting of triphenylphosphonium
halides, trialkylphosphonium halides, tricycloalkylphosphonium
halides or alkyl-phosphonates, and secondly a catalyst which may be
selected from the group consisting of alkoxy-alkaline metals (e.g.
wherein the metal is Li, Na or K), DBU, DBN and guanidine.
[0124] Then in reaction step (e), this compound is reacted with an
halogen, preferably chlorine or bromine, in the presence of a
chlorinated solvent such as carbon tetrachloride or chloroform.
Finally in reaction step (f), the product of step (e) is reacted
with a catalyst which may be selected from the group consisting of
alkoxy-alkaline metals (e.g. wherein the metal is Li, Na or K),
DBU, DBN and guanidine.
[0125] FIG. 5 represents a scheme for performing various
alterations of a substituent R.sub.3 starting from a known
substituted lumazine wherein the said R.sub.3 is either formyl. In
reaction step (a), a 6-formyl substituted lumazine is reacted with
a compound having the formula R.sub.5CH.sub.2NO.sub.2 in the
presence of a base (such as a tertiary amine) and a protic or
aprotic solvent. Then in step (b), the product of step (a) may be
reacted with acetic anhydride in the presence of pyridine and a
base in an aprotic solvent. Then in step (e), the product of step
(b) may be reacted with hydrogen sulfide or a thiol R.sub.6SH
wherein R.sub.6 may be alkyl or aryl in a protic solvent.
Alternatively in step (c), the starting 6-formyl substituted
lumazine is reacted with a compound selected from the group
consisting of R.sub.7-oxycarbonyl-methyltriphenylphosphonium
halides, R.sub.7-oxycarbonyl-methyltrialkylphosphonium halides,
R.sub.7oxycarbonylmethylalkyl-phosphonates and the corresponding
arsenium compounds in the presence of a base. Then in step (d), the
product of step (c) may be reacted with an halogen, preferably
chlorine or bromine, or an N-halosuccinimide in the presence of a
protic solvent. Then in step (f), the product of step (d) may be
reacted with a catalyst such as DBU, DBN or guanidine in an aprotic
solvent in order to produce one or more halogenated isomers wherein
the halogen atom X may be on one or the other side of the double
bond. Then in step (g), the product of step (f) may be reacted with
a compound having the formula R.sub.8OM (wherein M is a metal such
as Na, K or Li and R.sub.8 may be an alkoxy, alkoxylaryl or
alkoxyheterocyclic radical).
[0126] FIG. 6 represents a scheme for preparing a substituted
lumazine wherein the said R.sub.4 is either arylalkyl or hydroxyl
and performing various alterations of a substituent R.sub.4
starting from the said 7-hydroxyl substituted lumazine. In step
(a), a 6-amino-5-nitroso-pyrimidin-2,4-dione being optionally
substituted in the 1-position (with substituent R.sub.2) and/or in
the 3-position (with substituent R.sub.1) is hydrogenated via
reaction with: [0127] either hydrogen in the presence of a
hydrogenation catalyst (such as PtO.sub.2) in the presence of a
protic solvent, [0128] or sodium dithionite in the presence of
water, in order to obtain an optionally substituted
5,6-diamino-uracil.
[0129] In step (b), the said optionally substituted
5,6-diamino-uracil is reacted with an arylmethylglyoxal in the
presence of a protic solvent in order to obtain a 7-arylmethyl
lumazine being optionally further substituted in position 1 and/or
in position 3 of the pteridine ring. In step (c), the latter is
further modified into the corresponding (optionally substituted)
7-acyl lumazine (e.g. a 7-benzoyllumazine) through an oxidation
reaction for instance by means of KMnO.sub.4 in the presence of
water.
[0130] Alternatively, the optionally substituted 5,6-diamino-uracil
from step (a) is transformed into a 7-hydroxy lumazine being
substituted in the 6-position with substituent R.sub.3 through the
combined steps (d) and (e) of: [0131] reaction with an alkyl
arylglyoxylate or alkyl alkylglyoxylate or alkyl heterocyclic
glyoxylate or alkyl arylalkylglyoxylate or alkyl heterocyclic
alkylglyoxylate in the presence of a protic solvent, and [0132]
acidification by means of an acid (e.g. nitric, sulfuric,
chlorhydric acids and the like) in the presence of water.
[0133] The hydroxyl substituent of the R.sub.3-substituted lumazine
from step (e) may then be replaced by a thiol, mercapto, alkoxy,
chloro or amino substituent by any of steps (f), (g), (h) and (l)
or any combination of such steps. For instance, in step (f) the
R.sub.3-substituted lumazine from step (e) is reacted with
P.sub.4S.sub.10 in the presence of pyridine. In step (f) the
R.sub.3-substituted lumazine from step (e) or thiol analogue from
step (f) is reacted with either a dialkylsulfate or a compound
having the formula XR.sub.5 wherein X may be halogen, tosyl, mesyl
and the like, and R.sub.5 may be alkyl, arylalkyl,
heterocycloalkyl, alkoxycarbonylalkyl or aryloxycarbonyl-alkyl. In
step (h), the R.sub.3-substituted lumazine from step (e) is reacted
with POCl.sub.3 in the presence of ammonium chloride. In step (i)
the 7-chloro lumazine from step (h) is reacted, in the presence of
an aprotic solvent, with an amine, alcohol, phenol, thiol or
thiophenol having the formula HR.sub.6 (wherein R.sub.6 may be
selected from the group consisting of amino, alkylamino,
cycloalkylamino, arylamino, heterocyclo-alkylamino, alkoxy,
aryloxy, alkylaryloxy, arylalkyloxy, heterocycloalkyloxy, thio,
alkylthio, arylthio, arylalkylthio, alkylarylthio,
heterocycloalkylthio) in the further presence of CsF and a catalyst
such as a crown ether (e.g. 18-crown-6).
[0134] FIG. 7 represents a general scheme for preparing first a
poly-substituted lumazine bearing either a multiplet of
substituents (R.sub.1, R.sub.3, R.sub.4) or a multiplet of
substituents (R.sub.2, R.sub.3, R.sub.4) and then introducing a
further substituent (R.sub.2 or R.sub.1 respectively) into the
compound of the first step. The first step consists of two
sub-steps (a, b) which may be performed either simultaneously or
subsequently. In the first sub-step (a), an uracil bearing a
substituent R.sub.2 or R.sub.1, wherein R.sub.1 and R.sub.2 may be
selected from the group consisting of hydrogen, alkyl, cycloalkyl,
arylalkyl and heterocycloalkyl, is reacted in a protic solvent with
a reactant bearing atoms or groups R.sub.3 and R.sub.4, wherein
R.sub.3, R.sub.4 may be selected from the group consisting of
hydrogen, hydroxyl, alkyl, cycloalkyl, aryl, arylalkyl, alkylaryl
and heterocyclic, and wherein the said reactant may be selected
from the group consisting of: [0135] alkyl arylglyoxylates, alkyl
alkylglyoxylates, alkyl heterocyclic glyoxylates, alkyl
arylalkylglyoxylates, and alkyl heterocyclic alkylglyoxylates, in
which case R.sub.4 is hydroxyl; [0136] alkylglyoxals, arylglyoxals,
alkylarylglyoxals, arylalkylglyoxals, heterocyclic glyoxals, and
the monoximes corresponding to the said glyoxals, in which case one
of R.sub.3 and R.sub.4 is hydrogen; and [0137]
dialkylethylenediones, alkyl arylethylenediones,
diarylethylenediones, alkyl heterocyclic ethylenediones and
diheterocyclic ethylenediones, in which case R.sub.3 and R.sub.4
are both different from hydrogen and hydroxyl. In the second
sub-step (b), reaction is completed by acidification of the
reaction mixture.
[0138] After the first step (a, b) is completed, the resulting
poly-substituted lumazine bearing either a multiplet of
substituents (R.sub.1, R.sub.3, R.sub.4) or a multiplet of
substituents (R.sub.2, R.sub.3, R.sub.4) is reacted, through step
(c) or step (d) respectively, in the presence of a base and a polar
aprotic solvent or a protic solvent, with a reactant R.sub.2X or
R.sub.1X respectively (wherein R.sub.1 and R.sub.2 may each be
selected from the group consisting of alkyl, cycloalkyl, arylalkyl
and heterocycloalkyl, and X may be selected from the group
consisting of chloro, bromo, iodo, tosylate, mesylate and the
like).
[0139] When R.sub.4 and R.sub.3 together form an optionally
substituted aryl radical, this is achieved by aromatic cyclization
techniques well known in the art.
[0140] Further methods of preparation are shown in FIGS. 8 to 11
and are further explained and illustrated in the following
exmples.
[0141] When applicable, and depending upon the specific
substituents being present, not only the novel poly-substituted
pteridine-2,4-diones (lumazines), and mono- and polysubstituted
2-thiolumazines, 4-thiolumazines and 2,4-dithiolumazines having the
general formula (I) but also the (thio)lumazines previously known
in the art without any indication of biological activity, i.e. all
of the (thio)lumazines having the general formula (IV) according to
this invention, may be in the form of a pharmaceutically acceptable
salt. The latter include any therapeutically active non-toxic
addition salts which compounds having the general formula (I) are
able to form with a salt-forming agent. Such addition salts may
conveniently be obtained by treating any of the (thio)lumazine
compounds of the invention having the general formula (I), or the
general formula (IV), with an appropriate salt-forming acid or
base. Examples of such appropriate salt-forming acids include, for
instance, inorganic acids resulting in forming salts such as but
not limited to the hydrochloride, hydrobromide, sulfate, nitrate,
phosphate, diphosphate, bicarbonate, carbonate, and the like, of
(thio)lumazine compounds having the general formula (I), or the
general formula (IV); or organic monocarboxylic or dicarboxylic
acids resulting in forming salts such as, for example, the acetate,
propanoate, hydroxyacetate, 2-hydroxypropanoate, 2-oxopropanoate,
lactate, pyruvate, oxalate, malonate, succinate, maleate, fumarate,
malate, tartrate, citrate, methanesulfonate, ethanesulfonate,
benzoate, benzenesulfonate, p-oluene-sulfonate, salicylate,
p-aminosalicylate, pamoate, bitartrate, camphorsulfonate, edetate,
1,2-ethanedisulfonate, fumarate, glucoheptonate, gluconate,
glutamate, hexylresorcinate, hydroxynaphtoate,
hydroxyethanesulfonate, lactate, mandelate, methylsulfate,
pantothenate, stearate and the like, of (thio)lumazine compounds
having the general formula (I), or the general formula (IV).
Examples of appropriate salt-forming bases include, for instance,
inorganic bases like metallic hydroxides such as but not limited to
those of calcium, lithium, magnesium, potassium, sodium and zinc,
resulting in the corresponding metal salt of the (thio)lumazine
compounds having the general formula (I), or the general formula
(IV); organic bases such as but not limited to
N,N'-dibenzylethylenediamine, chloroprocaine, choline,
diethanolamine, ethylene-diamine, N-methylglucamine, procaine and
the like.
[0142] Reaction conditions for treating the (thio)lumazine
compounds of this invention having the general formula (I), or the
general formula (IV), with an appropriate salt-forming acid or base
are similar to standard conditions involving the same acid or base
but different compounds to be submitted to salification. Preferably
the pharmaceutically acceptable salt will be selected so as to
impart greater water-solubility, lower toxicity, greater stability
and/or slower dissolution rate to the compounds of this invention.
The term "pharmaceutically acceptable salt" as used herein also
includes any solvate which may be formed with a suitable inorganic
solvent (e.g. hydrates) or organic solvent, such as but not limited
to alcohols, ketones, esters and the like.
[0143] The present invention further provides the use of a
poly-substituted pteridinedione (lumazine), as well as a mono- or
poly-substituted 2-thiolumazine, 4-thiolumazine or
2,4-dithiolumazine represented by the general formula (I), or the
general formula (IV), or a pharmaceutically acceptable salt
thereof, as a biologically-active ingredient, especially as a
medicine or a diagnostic agent or for the manufacture of a
medicament or a diagnostic kit. In particular the said medicament
may be for the prevention or treatment of a pathologic condition
selected from the group consisting of: [0144] immune disorders, in
particular organ and cells transplant rejections, and autoimmune
disorders, [0145] cardiovascular disorders, [0146] disorders of the
central nervous system, and [0147] cell proliferative
disorders.
[0148] The pathologic conditions and disorders concerned by the
said use, and the corresponding methods of prevention or treatment,
are detailed hereinbelow. Any of the uses mentioned with respect to
the present invention may be restricted to a non-medical use (e.g.
in a cosmetic composition), a non-therapeutic use, a non-diagnostic
use, a non-human use (e.g. in a veterinary composition), or
exclusively an in-vitro use, or a use with cells remote from an
animal.
[0149] The invention further relates to a pharmaceutical
composition comprising: [0150] (a) one or more tri- or
tetra-substituted pteridinediones (lumazines), 2-thiolumazines,
4-thiolumazines or 2,4-dithiolumazines represented by the general
formula (1), or the general formula (IV), and [0151] (b) one or
more pharmaceutically acceptable carriers.
[0152] In a third embodiment, this invention provides combinations,
preferably synergistic combinations, of one or more tri- or
tetra-substituted pteridinediones (lumazines), 2-thiolumazines,
4-thiolumazines or 2,4-dithiolumazines represented by the general
formula (I), or the general formula (IV), with one or more
biologically-active drugs being preferably selected from the group
consisting of immunosuppressant and/or immunomodulator drugs,
antineoplastic drugs, and antiviral agents. As is conventional in
the art, the evaluation of a synergistic effect in a drug
combination may be made by analyzing the quantification of the
interactions between individual drugs, using the median effect
principle described by Chou et al. in Adv. Enzyme Reg. (1984)
22:27. Briefly, this principle states that interactions (synergism,
additivity, antagonism) between two drugs can be quantified using
the combination index (hereinafter referred as CI) defined by the
following equation: CI x = ED x 1 .times. c ED x 1 .times. a + ED x
2 .times. c ED x 2 .times. a ##EQU1## wherein ED.sub.x is the dose
of the first or respectively second drug used alone (1a, 2a), or in
combination with the second or respectively first drug (1c, 2c),
which is needed to produce a given effect. The said first and
second drug have synergistic or additive or antagonistic effects
depending upon CI<1, CI=1, or CI>1, respectively. As will be
explained in more detail herein-below, this principle may be
applied to a number of desirable effects such as, but not limited
to, an activity against transplant rejection, an activity against
immunosuppression or immunomodulation, or an activity against cell
proliferation.
[0153] For instance the present invention relates to a
pharmaceutical composition or combined preparation having
synergistic effects against immunosuppression or immunomodulation
and containing: [0154] (a) one or more immunosuppressant and/or
immunomodulator drugs, and [0155] (b) at least one poly-substituted
pteridinedione (lumazine), or mono- or poly-substituted
2-thiolumazine, 4-thiolumazine or 2,4-dithiolumazine represented by
the general formula (1), or the general formula (IV), and [0156]
(c) optionally one or more pharmaceutical excipients or
pharmaceutically acceptable carriers, for simultaneous, separate or
sequential use in the treatment or prevention of autoimmune
disorders and/or in transplant-rejections.
[0157] The advantages to associate a (thio)lumazine compound
represented by the general formula (I), or the general formula
(IV), with one or more other immunosuppressants are that: [0158]
the therapeutic spectrum of action of the individual components is
quantitatively and qualitatively broadened, and [0159] it allows,
by means of a dose reduction without reduced efficacy but with
increased safety, the treatment of immune disorders which hitherto
had no indication for immunosuppressive therapy as a result of side
effects. At the same time, the therapy costs can be decreased to an
appreciable extent.
[0160] Suitable immunosuppressant drugs for inclusion in the
synergistic compositions or combined preparations of this invention
are preferably selected from the group consisting of cyclosporin A,
substituted xanthines (e.g. methylxanthines such as
pentoxyfylline), tacrolimus, rapamycin (and derivatives thereof),
leflunomide (or its main active metabolite A771726, or analogs
thereof called malononitrilamides), mycophenolic acid and salts
thereof (including the sodium salt marketed under the trade name
Mofetil.RTM.), adrenocortical steroids, azathioprine, brequinar,
gusperimus, 6-mercaptopurine, mizoribine, chloroquine,
hydroxychloroquine and monoclonal antibodies with immunosuppressive
properties. Adrenocortical steroids within the meaning of this
invention mainly include glucocorticoids such as but not limited to
dexamethasone, methylprednisolone, methotrexate, prednisone,
prednisolone, triamcinolone and pharmaceutically acceptable salts
thereof. Rapamycin derivatives as referred herein include
O-alkylated derivatives, particularly 9-deoxorapamycins,
26-dihydrorapamycins, 40-O-substituted rapamycins and
28,40-O,O-disubstituted rapamycins (as disclosed in U.S. Pat. No.
5,665,772) such as 40-O-(2-hydroxy)ethyl rapamycin--also known as
SDZ-RAD-, pegylated rapamycin (as disclosed in U.S. Pat. No.
5,780,462), ethers of 7-desmethylrapamycin (as disclosed in U.S.
Pat. No. 6,440,991) and polyethylene glycol esters of SDZ-RAD (as
disclosed in U.S. Pat. No. 6,331,547).
[0161] Suitable immunomodulator drugs for inclusion into the
synergistic immunomodulating pharmaceutical compositions or
combined preparations of this invention are preferably selected
from the group consisting of acemannan, amiprilose, bucillamine,
ditiocarb sodium, imiquimod, Inosine Pranobex, interferon-.beta.,
interferon-.gamma., lentinan, levamisole, pidotimod, romurtide,
platonin, procodazole, propagermanium, thymomodulin, thymopentin
and ubenimex.
[0162] Synergistic activity of the pharmaceutical compositions or
combined preparations of this invention against immunosuppression
or immunomodulation may be readily determined by means of one or
more tests such as, but not limited to, the MLR (abbreviation
standing for "mixed lymphocyte reaction") test, or a test wherein
TNF-.alpha. or IL-1.beta. inhibition and/or a test wherein the
activation of a cluster of differentiation (hereinafter referred as
CD) is quantified. The synergistic effect may be evaluated by the
median effect analysis method described herein-before. Such tests
may for instance, according to standard practice in the art,
involve the use of equiment, such as flow cytometer, being able to
separate and sort a number of cell subcategories at the end of the
analysis, before these purified batches can be analyzed
further.
[0163] Synergistic activity of the pharmaceutical compositions of
this invention in the treatment of transplant rejection may be
readily determined by means of one or more tests such as but not
limited to the Whole Blood Assay (hereinafter referred as WBA) as
described for instance by Lin et al. in Transplantation (1997)
63:1734-1738. WBA is a lymphoproliferation assay performed in vitro
using lymphocytes present in the whie blood, taken from animals
that were previously given test substances in vivo. Hence it
reflects the in vivo effect of substances as assessed by an in
vitro read-out assay. The synergistic effect is evaluated by the
median effect analysis method described herein-before.
[0164] The pharmaceutical composition or combined preparation with
synergistic activity against immunosuppression or immunomodulation
according to this invention may contain the compound of general
formula (I), or the general formula (IV), over a broad content
range depending on the contemplated use and the expected effect of
the preparation. Generally, the (thio)lumazine content of the
combined preparation is within the range of 0.1 to 99.9% by weight,
preferably from 1 to 99% by weight, more preferably from 5 to 95%
by weight.
[0165] The invention further relates to a composition or combined
preparation having synergistic effects against cell proliferation
and containing: [0166] (a) one or more antineoplastic drugs, and
[0167] (b) at least one poly-substituted pteridinedione (lumazine),
or a mono- or poly-substituted 2-thiolumazine, 4-thiolumazine or
2,4-dithiolumazine represented by the general formula (I), or the
general formula (IV), and [0168] (c) optionally one or more
pharmaceutical excipients or pharmaceutically acceptable carriers,
for simultaneous, separate or sequential use in the treatment or
prevention of cell proliferative disorders.
[0169] Suitable antineoplastic drugs for inclusion into the
synergistic antiproliferative pharmaceutical compositions or
combined preparations of this invention are preferably selected
from the group consisting of alkaloids, alkylating agents
(including but not limited to alkyl sulfonates, aziridines,
ethylenimines, methylmelamines, nitrogen mustards and
nitrosoureas), antibiotics, antimetabolites (including but not
limited to folic acid analogs, purine analogs and pyrimidine
analogs), enzymes, interferon and platinum complexes.
[0170] Synergistic activity of the pharmaceutical compositions or
combined preparations of this invention against cell proliferation
may be readily determined by means of one or more tests such as,
but not limited to, the measurement of the radioactivity resulting
from the incorporation of .sup.3H-thymidine in culture of tumor
cell lines. For instance, different tumor cell lines are selected
in order to evaluate the anti-tumor effects of the test compounds,
such as but not limited to: [0171] RPMI1788: human Peripheral Blood
Leucocytes (PBL) Caucasian tumor line, [0172] Jurkat: human acute T
cell leukemia, [0173] EL4: C57BI/6 mouse lymphoma, or [0174] THP-1:
human monocyte tumor line. Depending on the selected tumor cell
line, different culture media may be used, such as for example:
[0175] for RPMI1788 and THP-1: RPMI-1640+10% FCS+1% NEM+1% sodium
pyruvate+5.times.10.sup.-5 mercapto-ethanol+antibiotics (G-418 0.45
.mu.g/ml). [0176] for Jurkat and EL4: RPMI-1640+10% FCS+antibiotics
(G-418 0.45 .mu.g/ml).
[0177] In a specific embodiment of the synergy determination test,
the tumor cell lines are harvested and a suspension of
0.27.times.10.sup.6 cells/ml in whole medium is prepared. The
suspensions (150 .mu.l) are added to a microtiter plate in
triplicate. Either complete medium (controls) or the test compounds
at the test concentrations (50 .mu.l) are added to the cell
suspension in the microtiter plate. The cells are incubated at
37.degree. C. under 5% CO.sub.2 for about 16 hours.
.sup.3H-thymidine is added, and the cells incubated for another 8
hours. The cells are harvested and radioactivity is measured in
counts per minute (CPM) in a .beta.-counter. The .sup.3H-thymidine
cell content, and thus the measured radioactivity, is proportional
to the proliferation of the cell lines. The synergistic effect is
evaluated by the median effect analysis method as disclosed
herein-before.
[0178] The pharmaceutical composition or combined preparation with
synergistic activity against cell proliferation according to this
invention may contain the (thio)lumazine compound having the
general formula (I), or the general formula (IV), over a broad
content range depending on the contemplated use and the expected
effect of the preparation. Generally, the (thio)lumazine content of
the combined preparation is within the range of 0.1 to 99.9% by
weight, preferably from 1 to 99% by weight, more preferably from 5
to 95% by weight.
[0179] The invention further relates to a pharmaceutical
composition or combined preparation having synergistic effects
against a viral infection and containing: [0180] (a) one or more
anti-viral agents, and [0181] (b) at least one poly-substituted
pteridinedione (lumazine), or mono- or poly-substituted
2-thiolumazine, 4-thiolumazine or 2,4dithiolumazine represented by
the general formula (I), or the general formula (IV), and [0182]
(c) optionally one or more pharmaceutical excipients or
pharmaceutically acceptable carriers, for simultaneous, separate or
sequential use in the treatment or prevention of a viral
infection.
[0183] Suitable anti-viral agents for inclusion into the
synergistic antiviral compositions or combined preparations of this
invention include, for instance, retroviral enzyme inhibitors
belonging to categories well known in the art, such as HIV-1 IN
inhibitors, nucleoside reverse transcriptase inhibitors (e.g.
zidovudine, lamivudine, didanosine, stavudine, zalcitabine and the
like), non-nucleoside reverse transcriptase inhibitors (e.g.
nevirapine, delavirdine and the like), other reverse transcriptase
inhibitors (e.g. foscarnet sodium and the like), and HIV-1 protease
inhibitors (e.g. saquinavir, ritonavir, indinavir, nelfinavir and
the like). Other suitable antiviral agents include for instance
acyclovir, cidofovir, cytarabine, edoxudine, famciclovir,
floxuridine, ganciclovir, idoxuridine, penciclovir, sorivudine,
trifluridine, valaciclovir, vidarabine, kethoxal, methisazone,
moroxydine, podophyllotoxin, ribavirine, rimantadine, stallimycine,
statolon, tromantadine and xenazoic acid.
[0184] Especially relevant to this aspect of the invention is the
inhibition of the replication of viruses selected from the group
consisting of picoma-, toga-, bunya-, orthomyxo-, paramyxo-,
rhabdo-, retro-, arena-, hepatitis B-, hepatitis C-, hepatitis D-,
adeno-, vaccinia-, papilloma-, herpes-, corona-, varicella- and
zoster-virus, in particular human immunodeficiency virus (HIV).
Synergistic activity of the pharmaceutical compositions or combined
preparations of this invention against viral infection may be
readily determined by means of one or more tests such as, but not
limited to, the isobologram method, as previously described by
Elion et al. in J. Biol. Chem. (1954) 208:477-488 and by Baba et
al. in Antimicrob. Agents Chemother. (1984) 25:515-517, using
EC.sub.50 for calculating the fractional inhibitory concentration
(hereinafter referred as FIC). When the minimum FIC index
corresponding to the FIC of combined compounds (e.g.,
FIC.sub.x+FIC.sub.y) is equal to 1.0, the combination is said to be
additive; when it is beween 1.0 and 0.5, the combination is defined
as subsynergistic, and when it is lower than 0.5, the combination
is by defined as synergistic. When the minimum FIC index is between
1.0 and 2.0, the combination is defined as subantagonistic and,
when it is higher than 2.0, the combination is defined as
antagonistic.
[0185] The pharmaceutical composition or combined preparation with
synergistic activity against viral infection according to this
invention may contain the (thio)lumazine compound having the
general formula (I), or the general formula (IV), over a broad
content range depending on the contemplated use and the expected
effect of the preparation. Generally, the (thio)lumazine content of
the combined preparation is within the range of 0.1 to 99.9% by
weight, preferably from 1 to 99% by weight, more preferably from 5
to 95% by weight.
[0186] The pharmaceutical compositions and combined preparations
according to this invention may be administered orally or in any
other suitable fashion. Oral administration is preferred and the
preparation may have the form of a tablet, aqueous dispersion,
dispersable powder or granule, emulsion, hard or soft capsule,
syrup, elixir or gel. The dosing forms may be prepared using any
method known in the art for manufacturing these pharmaceutical
compositions and may comprise as additives sweeteners, flavoring
agents, coloring agents, preservatives and the like. Carrier
materials and excipients are detailed hereinbelow and may include,
inter alia, calcium carbonate, sodium carbonate, lactose, calcium
phosphate or sodium phosphate; granulating and disintegrating
agents, binding agents and the like. The pharmaceutical composition
or combined preparation may be included in a gelatin capsule mixed
with any inert solid diluent or carrier material, or has the form
of a soft gelatin capsule, in which the ingredient is mixed with a
water or oil medium. Aqueous dispersions may comprise the
biologically active composition or combined preparation in
combination with a suspending agent, dispersing agent or wetting
agent. Oil dispersions may comprise suspending agents such as a
vegetable oil. Rectal administration is also applicable, for
instance in the form of suppositories or gels. Injection is also
applicable as a mode of administration, for instance in the form of
injectable solutions or dispersions.
[0187] Auto-immune disorders to be prevented or treated by the
pharmaceutical compositions or combined preparations of this
invention include both systemic auto-immune diseases such as but
not limited to lupus erythematosus, psoriasis, vasculitis,
polymyositis, scleroderma, multiple sclerosis, ankylosing
spondilytis, rheumatoid arthritis and Sjogren syndrome; auto-immune
endocrine disorders such as thyroiditis; and organ-specific
auto-immune diseases such as but not limited to Addison disease,
hemolytic or pernicious anemia, Goodpasture syndrome, Graves
disease, idiopathic thrombocytopenic purpura, insulin-dependent
diabetes mellitus, juvenile diabetes, uveitis, Crohn's disease,
ulcerative colitis, pemphigus, atopic dermatitis, autoimmune
hepatitis, primary biliary cirrhosis, autoimmune pneumonitis,
auto-immune carditis, myasthenia gravis, glomerulonephritis and
spontaneous infertility.
[0188] Transplant rejections to be prevented or treated by the
pharmaceutical compositions or combined preparations of this
invention include the rejection of transplanted or grafted organs
or cells (both allografts and xenografts), such as but not limited
to host versus graft reaction and, especially after bone marrow
transplantation, graft versus host reaction or disease. "Organ"
herein means all organs or parts of organs in mammals, in
particular humans, such as but not limited to kidney, lung, bone
marrow, hair, cornea, eye (vitreous), heart, heart valve, liver,
pancreas, blood vessel, skin, muscle, bone, intestine or stomach.
"Rejection" as used herein mean all reactions of the recipient body
or of the transplanted organ which in the end lead to cell or
tissue death in the transplanted organ or adversely affect the
functional ability and viability of the transplanted organ or the
recipient. In particular, this means acute and chronic rejection
reactions. Also included in this invention is preventing or
treating the rejection of cell transplants and xenotransplantation.
The major hurdle for xenotransplantation is that even before the T
lymphocytes, responsible for the rejection of allografts, are
activated, the innate immune system, especially T-independent B
lymphocytes and macrophages are activated. This provokes two types
of severe and early acute rejection called hyper-acute rejection
and vascular rejection, respectively. The present invention
addresses the problem that conventional immunosuppressant drugs
like cyclosporin A are ineffective in xenotransplantation. The
ability of the compounds of this invention to suppress
T-independent xeno-antibody production as well as macrophage
activation may be evaluated in the ability to prevent xenograft
rejection in athymic, T-deficient mice receiving xenogenic
hamster-heart grafts.
[0189] Cell proliferative disorders to be prevented or treated by
the pharmaceutical compositions or combined preparations of this
invention include not only tumor progression or invasion or
metastasis inhibition of a cancer such as, but not limited to, a
cancer selected from the group consisting of lung cancer,
leukaemia, ovarian cancer, sarcoma, Kaposi's sarcoma, meningioma,
colon cancer, lymp node tumor, glioblastoma multiforme, prostate
cancer or skin carcinose, but also side effects associated with
current cancer therapies, including chemotherapy or radiation
therapy, such as gastrointestinal mucosal damage or
radiation-induced mucositis, the treatment being based on enhancing
resistance of mesenchymal cells to TNF.
[0190] CNS disorders to be prevented or treated by the
pharmaceutical compositions of this invention include cognitive
pathologies such as dementia, cerebral ischemia, trauma, epilepsy,
schizophrenia, chronic pain and neurologic disorders such as but
not limited to depression, social phobia and obsessive compulsive
disorders.
[0191] Cardiovascular disorders to be prevented or treated by the
pharmaceutical compositions of this invention include ischemic
disorders, infarct or reperfusion damage, atherosclerosis and
stroke.
[0192] The term "pharmaceutically acceptable carrier or excipient"
as used herein in relation to pharmaceutical compositions and
combined preparations means any material or substance with which
the biologically-active ingredient(s), i.e. the (thio)lumazine
having the general formula (1), or the general formula (IV), and
optionally the immunosuppressant or immunomodulator or
antineoplastic drug or antiviral agent, may be formulated in order
to facilitate its application or dissemination to the locus to be
treated, for instance by dissolving, dispersing or diffusing the
said composition, and/or to facilitate its storage, transport or
handling without impairing its effectiveness. The pharmaceutically
acceptable carrier may be a solid or a liquid or a gas which has
been compressed to form a liquid, i.e. the compositions of this
invention can suitably be used as concentrates, emulsions,
solutions, granulates, dusts, sprays, aerosols, pellets or
powders.
[0193] Suitable pharmaceutical carriers for use in the said
pharmaceutical compositions and their formulation are well known to
those skilled in the art. There is no particular restriction to
their selection within the present invention although, due to the
usually low or very low water-solubility of the poly-substituted
pteridinediones (lumazines), or mono- or poly-substituted
2-thiolumazines, 4-thiolumazines and 2,4-dithiolumazines of this
invention, special attention will be paid to the selection of
suitable carrier combinations that can assist in properly
formulating them in view of the expected time release profile.
Suitable pharmaceutical carriers include additives such as wetting
agents, dispersing agents, stickers, adhesives, emulsifying or
surface-active agents, thickening agents, complexing agents,
gelling agents, solvents, coatings, antibacterial and antifungal
agents (for example phenol, sorbic acid, chlorobutanol), isotonic
agents (such as sugars or sodium chloride) and the like, provided
the same are consistent with pharmaceutical practice, i.e. carriers
and additives which do not create permanent damage to mammals. The
pharmaceutical compositions of the present invention may be
prepared in any known manner, for instance by homogeneously mixing,
dissolving, spray-drying, coating and/or grinding the active
ingredients, in a one-step or a multi-steps procedure, with the
selected carrier material and, where appropriate, the other
additives such as surface-active agents may also be prepared by
micronisation, for instance in view to obtain them in the form of
microspheres usually having a diameter of about 1 to 10 .mu.m,
namely for the manufacture of microcapsules for controlled or
sustained release of the biologically active ingredient(s).
[0194] Suitable surface-active agents to be used in the
pharmaceutical compositions of the present invention are non-ionic,
cationic and/or anionic materials having good emulsifying,
dispersing and/or wetting properties. Suitable anionic surfactants
include both water-soluble soaps and water-soluble synthetic
surface-active agents. Suitable soaps are alkaline or
alkaline-earth metal salts, unsubstituted or substituted ammonium
salts of higher fatty acids (C.sub.10-C.sub.22), e.g. the sodium or
potassium salts of oleic or stearic acid, or of natural fatty acid
mixtures obtainable form coconut oil or tallow oil. Synthetic
surfactants include sodium or calcium salts of polyacrylic acids;
fatty sulphonates and sulphates; sulphonated benzimidazole
derivatives and alkylarylsulphonates. Fatty sulphonates or
sulphates are usually in the form of alkaline or alkaline-earth
metal salts, unsubstituted ammonium salts or ammonium salts
substituted with an alkyl or acyl radical having from 8 to 22
carbon atoms, e.g. the sodium or calcium salt of lignosulphonic
acid or dodecylsulphonic acid or a mixture of fatty alcohol
sulphates obtained from natural fatty acids, alkaline or
alkaline-earth metal salts of sulphuric or sulphonic acid esters
(such as sodium lauryl sulphate) and sulphonic acids of fatty
alcohol/ethylene oxide adducts. Suitable sulphonated benzimidazole
derivatives preferably contain 8 to 22 carbon atoms. Examples of
alkylarylsulphonates are the sodium, calcium or alcanolamine salts
of dodecylbenzene sulphonic acid or dibutyl-naphtalenesulphonic
acid or a naphtalene-sulphonic acid/formaldehyde condensation
product. Also suitable are the corresponding phosphates, e.g. salts
of phosphoric acid ester and an adduct of p-nonylphenol with
ethylene and/or propylene oxide, or phospholipids. Suitable
phospholipids for this purpose are the natural (originating from
animal or plant cells) or synthetic phospholipids of the cephalin
or lecithin type such as e.g. phosphatidylethanolamine,
phosphatidylserine, phosphatidylglycerine, lysolecithin,
cardiolipin, dioctanylphosphatidylcholine,
dipalmitoylphoshatidylcholine and their mixtures.
[0195] Suitable non-ionic surfactants include polyethoxylated and
polypropoxylated derivatives of alkylphenols, fatty alcohols, fatty
acids, aliphatic amines or amides containing at least 12 carbon
atoms in the molecule, alkylarenesulphonates and
dialkylsulphosuccinates, such as polyglycol ether derivatives of
aliphatic and cycloaliphatic alcohols, saturated and unsaturated
fatty acids and alkylphenols, said derivatives preferably
containing 3 to 10 glycol ether groups and 8 to 20 carbon atoms in
the (aliphatic) hydrocarbon moiety and 6 to 18 carbon atoms in the
alkyl moiety of the alkylphenol. Further suitable non-ionic
surfactants are water-soluble adducts of polyethylene oxide with
poylypropylene glycol, ethylenediaminopolypropylene glycol
containing 1 to 10 carbon atoms in the alkyl chain, which adducts
contain 20 to 250 ethyleneglycol ether groups and/or 10 to 100
propyleneglycol ether groups. Such compounds usually contain from 1
to 5 ethyleneglycol units per propyleneglycol unit. Representative
examples of non-ionic surfactants are nonylphenolpolyethoxyethanol,
castor oil polyglycolic ethers, polypropylene/polyethylene oxide
adducts, tributylphenoxypolyethoxyethanol, polyethyleneglycol and
octylphenoxypolyethoxyethanol. Fatty acid esters of polyethylene
sorbitan (such as polyoxyethylene sorbitan trioleate), glycerol,
sorbitan, sucrose and pentaerythritol are also suitable non-ionic
surfactants.
[0196] Suitable cationic surfactants include quaternary ammonium
salts, preferably halides, having 4 hydrocarbon radicals optionally
substituted with halo, phenyl, substituted phenyl or hydroxy; for
instance quaternary ammonium salts containing as N-substituent at
least one C.sub.8-C.sub.22 alkyl radical (e.g. cetyl, lauryl,
palmityl, myristyl, oleyl and the like) and, as further
substituents, unsubstituted or halogenated lower alkyl, benzyl
and/or hydroxy-lower alkyl radicals.
[0197] A more detailed description of surface-active agents
suitable for this purpose may be found for instance in
"McCutcheon's Detergents and Emulsifiers Annual" (MC Publishing
Crop., Ridgewood, N.J., 1981), "Tensid-Taschenbuch", 2.sup.nd ed.
(Hanser Verlag, Vienna, 1981) and "Encyclopaedia of Surfactants
(Chemical Publishing Co., New York, 1981).
[0198] Structure-forming, thickening or gel-forming agents may be
included into the pharmaceutical compositions and combined
preparations of the invention. Suitable such agents are in
particular highly dispersed silicic acid, such as the product
commercially available under the trade name Aerosil; bentonites;
tetraalkyl ammonium salts of montmorillonites (e.g., products
commercially available under the trade name Bentone), wherein each
of the alkyl groups may contain from 1 to 20 carbon atoms;
cetostearyl alcohol and modified castor oil products (e.g. the
product commercially available under the trade name
Antisettle).
[0199] Gelling agents which may be included into the pharmaceutical
compositions and combined preparations of the present invention
include, but are not limited to, cellulose derivatives such as
carboxymethylcellulose, cellulose acetate and the like; natural
gums such as arabic gum, xanthum gum, tragacanth gum, guar gum and
the like; gelatin; silicon dioxide; synthetic polymers such as
carbomers, and mixtures thereof. Gelatin and modified celluloses
represent a preferred class of gelling agents.
[0200] Other optional excipients which may be included in the
pharmaceutical compositions and combined preparations of the
present invention include additives such as magnesium oxide; azo
dyes; organic and inorganic pigments such as titanium dioxide;
UV-absorbers; stabilisers; odor masking agents; viscosity
enhancers; antioxidants such as, for example, ascorbyl palmitate,
sodium bisulfite, sodium metabisulfite and the like, and mixtures
thereof; preservatives such as, for example, potassium sorbate,
sodium benzoate, sorbic acid, propyl gallate, benzylalcohol, methyl
paraben, propyl paraben and the like; sequestering agents such as
ethylene-diamine tetraacetic acid; flavoring agents such as natural
vanillin; buffers such as citric acid and acetic acid; extenders or
bulking agents such as silicates, diatomaceous earth, magnesium
oxide or aluminum oxide; densification agents such as magnesium
salts; and mixtures thereof.
[0201] Additional ingredients may be included in order to control
the duration of action of the biologically-active ingredient in the
compositions and combined preparations of the invention. Control
release compositions may thus be achieved by selecting appropriate
polymer carriers such as for example polyesters, polyamino-acids,
polyvinyl-pyrrolidone, ethylene-vinyl acetate copolymers,
methylcellulose, carboxymethylcellulose, protamine sulfate and the
like. The rate of drug release and duration of action may also be
controlled by incorporating the active ingredient into particles,
e.g. microcapsules, of a polymeric substance such as hydrogels,
polylactic acid, hydroxymethylcellulose, polymethyl methacrylate
and the other above-described polymers. Such methods include
colloid drug delivery systems like liposomes, microspheres,
microemulsions, nanoparticles, nanocapsules and so on. Depending on
the route of administration, the pharmaceutical composition or
combined preparation of the invention may also require protective
coatings.
[0202] Pharmaceutical forms suitable for injectable use include
sterile aqueous solutions or dispersions and sterile powders for
the extemporaneous preparation thereof. Typical carriers for this
purpose therefore include biocompatible aqueous buffers, ethanol,
glycerol, propylene glycol, polyethylene glycol, complexing agents
such as cyclodextrins and the like, and mixtures thereof.
[0203] Since, in the case of combined preparations including a
(thio)lumazine of this invention having the general formula (I), or
the general formula (IV), and another biologically-active drug such
as, but not limited to, an immunosuppressant or immunomodulator or
antineoplastic drug or antiviral agent, both active ingredients do
not necessarily bring out their synergistic therapeutic effect
directly at the same time in the patient to be treated, the said
combined preparation may be in the form of a medical kit or package
containing the two ingredients in separate but adjacent form. In
the latter context, each ingredient may therefore be formulated in
a way suitable for an administration route different from that of
the other ingredient, e.g. one of them may be in the form of an
oral or parenteral formulation whereas the other is in the form of
an ampoule for intravenous injection or an aerosol.
[0204] The present invention is useful for preventing or treating a
disease selected from the group consisting of CNS disorders, cell
proliferative disorders, viral infections, immune and auto-immune
disorders and transplant rejections in a subject or patient by
administering to the patient in need thereof an effective amount of
a (thio)lumazine compound having the general formula (I), or having
the general formula (I), or the general formula (IV), optionally
together with an effective amount of another biologically-active
drug such as, but not limited to, an immunosuppressant or
immunomodulator or antineoplastic drug or antiviral agent, or a
pharmaceutical composition such as disclosed above in extensive
details. The effective amount is usually in the range of 0.01 mg to
20 mg, preferably 0.1 mg to 5 mg, per day per kg bodyweight for
humans. Depending upon the pathologic condition to be treated and
the patient's condition, the said effective amount may be divided
into several sub-units per day or may be administered at more than
one day intervals.
[0205] The present invention further relates to the use of a
composition comprising: [0206] (a) one or more immunosuppressant
and/or immunomodulator drugs, and [0207] (b) at least one
(thio)lumazine represented by the general formula (I), or the
general formula (IV), in respective proportions such as to provide
a synergistic effect against immunosuppression or immunomodulation
in a human being. Similarly the invention relates to the use of a
composition comprising: [0208] (a) one or more immunosuppressant
and/or immunomodulator drugs, and [0209] (b) at least one
(thio)lumazine represented by the general formula (I), or the
general formula (IV), for the manufacture of a medicine for the
treatment of an immune or autoimmune disorder in a human being, in
respective proportions such as to provide a synergistic effect in
the said treatment.
[0210] The present invention further relates to a method for
selecting or classifying potent immunosuppressive agents,
particularly agents or drugs selected from the family of
(thio)lumazines represented by the general formula (I), or the
general formula (IV). Various models may be used for testing an
immunosuppressive effect. In vivo, for example, different
transplantation models are available. They are strongly influenced
by different immunogenicities, depending on the donor and recipient
species used and depending on the nature of the transplanted organ.
The survival time of transplanted organs can thus be used to
measure the suppression of the immune response.
[0211] In vitro, the most used models are lymphocyte activation
tests. Usually activation is measured via lymphocyte proliferation.
Inhibition of proliferation thus always means immunosuppression
under the experimental conditions applied. There exist different
stimuli for lymphocyte activation, in particular co-culture of
lymphocytes of different species (mixed lymphocyte reaction,
hereinafter referred as MLR) in a so-called mixed lymphocyte
culture test: lymphocytes expressing different minor and major
antigens of the HLA-DR type (i.e. alloantigens) activate each other
non-specifically.
[0212] The selection or classification method of this invenion is
based on the determination of a set of at least two lymphocyte
activation in vitro tests, wherein the said set includes at least
the MLR test and at least one other test selected from the group
consisting of a TNF-.alpha. assay and an IL-1 assay. Optionally the
selection or classification of agents or drugs of this invention
may be performed on the basis of more than two lymphocyte
activation in vitro tests, for instance on the basis of a set
further including an IL-6 assay or an IL-10 assay or an IL-12 assay
or an assay for a cluster of differentiation belonging to type I
transmembrane proteins such as but not limited to CD3 or CD28, or a
cluster belonging to type II transmembrane proteins such as, but
not limited to, CD69, CD71 or CD134.
[0213] Tables 2 and 7 below summarize some of the compounds of the
present invention that were made and tested for biological
activity, in particular lymphocyte activation, according to the
above-mentioned assays and test methods.
[0214] The following examples are provided only for illustration of
the invention and should in no way be understood as limiting its
scope.
EXAMPLES 1 TO 5
Preparation of Tri- and Tetra-Substituted
1,3-dimethyl-lumazines
[0215] The following tri- and tetra-substituted
1,3-dimethyl-lumazines were prepared according to methods and
procedures described hereinbefore by reference to FIGS. 6 and 7:
[0216] 6-[2-(p-trifluoromethylphenyl)ethenyl]-1,3-dimethyl-lumazine
(example 1), [0217]
6-[2-(p-trifluoromethylphenyl)ethenyl]-1,3-dimethyl-lumazine
(example 2), [0218] 6-[2-phenylethenyl]-1,3-dimethyl-lumazine
(example 3), [0219]
6-[2-(p-trifluoromethoxyphenyl)ethenyl]-1,3-dimethyl-lumazine
(example 4), and [0220]
6-cyano-7-ethylmercaptoacetate-1,3-dimethyl-lumazine (example
5).
EXAMPLE 6
Preparation of 3-methyl-6-phenyl-7-chloro-lumazine
[0221] 3-methyl-6-phenyl-7-chloro-lumazine was prepared according
to the general methods described hereinbefore by reference to FIG.
6.
EXAMPLES 7 TO 20
Preparation of 1,6-Disubstituted 7-hydroxylumazines
[0222] In a first step, trisubstituted lumazines having the general
formula (I) wherein R.sub.1 is hydrogen, R.sub.4 is hydroxyl, and
R.sub.2 and R.sub.3 are as indicated in table 1 below for each of
examples 7-13, were prepared according to the general methods
described hereinbefore by reference to FIG. 6, in particular steps
(d) and (e) thereof, and more specifically according to the
following procedure:
[0223] To a suspension of a 5,6-diamino-1-substituted uracil
hydrate (10 mmole) in 200 ml water, ethyl substituted
benzoylformate (12.5 mmole) was added. The resulting mixture was
heated under reflux for 40 minutes. After cooling to room
temperature, the precipitate was collected to yield a crude product
which was re-crystallized from a MeOH/water (1/1) mixture to yield
yellowish crystals or powder of the desired 7-hydroxyl
tri-substituted lumazine. TABLE-US-00001 TABLE 1 R.sub.2 R.sub.3
Example methyl phenyl 7 methyl p-methoxyphenyl 8 methyl p-toluyl 9
phenyl phenyl 10 benzyl phenyl 11 methyl m,p-dimethoxyphenyl 12
methyl p-chlorophenyl 13
In a second step, the following 7-chloro-1,6-disubstituted
lumazines: [0224] 1-methyl-6-phenyl-7-chloro-lumazine (example 14),
[0225] 1-methyl-6-(4'-methoxyphenyl)-7-chloro-lumazine (example
15), [0226] 1-methyl-6-(4'-methylphenyl)-7-chloro-lumazine (example
16), [0227] 1,6-diphenyl-7-chloro-lumazine (example 17), [0228]
1-benzyl-6-phenyl-7-chloro-lumazine (example 18), [0229]
1-methyl-6-(3',4'-dimethoxyphenyl)-7-chloro-lumazine (example 19),
and [0230] 1-methyl-6-(4'-chlorophenyl)-7-chloro-lumazine (example
20) were prepared according to the general methods described
hereinbefore by reference to FIG. 6, in particular step (h)
thereof, and more specifically according to the following
procedure:
[0231] To a suspension of 0.5 g NH.sub.4Cl in 20 ml POCl.sub.3, a
7-hydroxy tri-substituted lumazine of examples 7 to 13 (5 mmole)
was added. The resulted mixture was heated at 90.degree. C. till
the starting material completely disappeared. The reaction mixture
was concentrated under reduced pressure to a syrup and then 30 g
ice was added. After 30 minutes stirring at room temperature, the
precipitate was collected, washed with water and dried to yield a
crude product. The latter was then purified by chromatography on
silica gel (using MeOH/CH.sub.2Cl.sub.2 mixtures 1/100 to 1/20) to
yield the desired 7-chloro-1-substituted-6-substituted lumazine.
Crystals were obtained by re-crystallization from methanol.
[0232] Each compound of examples 14 to 20 was obtained in the
following yield and characterized by the following UV
(MeOH/H.sub.2O) and .sup.1H NMR (200 MHz, DMSO-d6) spectra:
[0233] Example 14: yield 54%; UV: 238.1, 274.7, 352.1 nm; .sup.1H
NMR: 12.1 (s, 1H), 7.74 (m, 2H), 7.56 (m, 3H) and 3.46 (s, 3H)
ppm.
[0234] Example 15: yield 40%; UV: 288.9, 356.9 nm; .sup.1H NMR:
12.1 (s, 1H), 8.06 (d, 2H), 7.04 (d, 2H), 3.82 (s, 3H) and 3.43 (s,
3H) ppm.
[0235] Example 16: yield 58%; UV: 279.5, 355.7 nm; .sup.1H NMR:
12.19 (s, 1H), 7.75 (d, 2H), 7.46 (d, 2H), 3.55 (s, 3H) and 2.50
(s, 3H) ppm.
[0236] Example 17: yield 60%; UV: 274.7, 349.7 nm; .sup.1H NMR:
12.25 (s, 1H), 7.73 (m, 2H), 7.54 (m, 5H), 7.43 (m, 3H) and 3.36
(s, 3H) ppm.
[0237] Example 18: yield 48%; UV: 274.7, 350.9 nm; .sup.1H NMR:
12.20 (s, 1H), 7.74 (m, 2H), 7.55 (m, 3H), 7.35 (m, 5H), 5.28 (s,
2H) and 3.45 (s, 3H) ppm.
[0238] Example 19: yield 62%; UV: 298.4, 365.0 nm; .sup.1H NMR:
12.07 (s, 1H), 7.35 (d, 1H), 7.30 (s, 1H), 7.12 (d, H), 3.83 (s,
3H), 3.80 (s, 3H) and 3.44 (s, 3H) ppm.
[0239] Example 20: yield 71%; UV: 238.1, 278.3 and 352.1 nm;
.sup.1H NMR: 12.13 (s, 1H), 7.77 (d, 2H), 7.64 (d, 2H) and 3.45 (s,
3H) ppm.
EXAMPLES 21 TO 25
Preparation of Tetra-Substituted 1,3-dimethyl-lumazines
[0240] The following tetra-substituted 1,3-dimethyl-lumazines:
[0241] 1,3-dimethyl-6-phenyl-7-phenoxy lumazine (example 21),
[0242] 1,3-dimethyl-6-phenyl-7-piperidino lumazine (example 22),
[0243] 1,3-dimethyl-6-phenyl-7-morpholino lumazine (example 23),
[0244] 1,3-dimethyl-6-phenyl-7-(4'-N-acetyl)piperazino lumazine
(example 24), and [0245]
1,3-dimethyl-6-phenyl-7-isopropylaminolumazine (example 25) were
prepared according to the general methods described hereinbefore by
reference to FIG. 6, in particular step (i) thereof, and more
specifically according to the following procedure:
[0246] To a mixture of 1.0 g of a 4 Angstrom molecular sieve, 200
mg CsF (1.2 mmole) in 4 ml THF, 40 mg 18-crown-6 (0.15 mmole) was
added. The resulting mixture was stirred at room temperature for
one hour. Then, 0.5 mmole of 6-phenyl-7-chloro-1,3-dimethyllumazine
and 0.6 mmole of a reactant HR.sub.6 (as defined in FIG. 6) were
added respectively (this reactant is phenol in example 21,
piperidine in example 22, morpholine in example 23,
4'-N-acetylpiperazine in example 24 and isopropylamine in example
25). The mixture was stirred at room temperature for another one
hour, and then filtrated through a pad of Celite.RTM.) (a filter
agent) and rinsed with CH.sub.2Cl.sub.2. After concentration under
reduced pressure, the residue was purified by chromatography on
silica gel (1-5% MeOH in CH.sub.2Cl.sub.2) to yield the desired
compound as a white or yellow powder.
[0247] Each compound of examples 21 to 25 was obtained in the
following yield and characterized by the following UV
(MeOH/H.sub.2O) and .sup.1H NMR (200 MHz, DMSO-d6 for examples
21-23, CDCl.sub.3 for examples 24 and 25) spectra:
[0248] Example 21: yield: 83%; UV: 283.0 and 348.5 nm; .sup.1H NMR:
8.11 (m, 2H), 7.53 (m, 5H), 7.38 (m, 3H) and 3.33 (s, 6H) ppm.
[0249] Example 22: yield: 99%; UV: 236.9, 308.0, and 374.6 nm;
.sup.1H NMR: 7.68 (m, 2H), 7.48 (m, 3H), 3.33 (s, 6H), 3.33 (m, 4H)
and 1.52 (m, 6H) ppm.
[0250] Example 23: yield: 99%; UV: 235.7, 304.4 and 367.4 nm;
.sup.1H NMR: 7.72 (m, 2H), 7.49 (m, 3H), 3.60 (m, 4H), 3.36 (m, 4H)
and 3.33 (s, 6H) ppm.
[0251] Example 24: yield: 98%; UV: 234.5, 303.2, 367.4 nm; .sup.1H
NMR: 7.77 (m, 2H), 7.45 (m, 3H), 3.66 (s, 1H), 3.51 (s, 3H),
3.40-3.70 (m, 8H) and 2.09 (s, 3H),
[0252] Example 25: yield: 99%; UV: 226.3, 293.7 and 354.5 nm;
.sup.1H NMR: 7.63 (m, 2H), 7.52 (m, 3H), 5.46 (br., 1 H), 4.32 (m,
1 H), 3.67 (s, 3H), 3.50 (s, 3H), 2.17 (s, 3H) and 1.28 (d,
6H).
EXAMPLE 26
In vitro Lymphocyte Activation Tests
[0253] All reagents were dissolved in 0.5 ml dimethylsulfoxide
(hereinafter referred as DMSO) and further diluted in culture
medium before use for the following in vitro experiments. The
commercially available culture medium consisted of RPMI-1640+10%
foetal calf serum (FCS).
[0254] Compounds described in some of the present examples were
tested in the following lymphocyte activation tests:
Mixed Lymphocyte Reaction
[0255] Peripheral blood mononuclear cells (hereinafter referred as
PBMC) were isolated from heparinized peripheral blood by density
gradient centrifugation over Lymphoprep (Nycomed, Maorstua,
Norway). Allogeneic PBMC or Eppstein-Barr Virus-transformed human B
cells [commercially available under the trade name RPMI1788 (ATCC
name CCL156)] which strongly express B7-1 and B7-2 antigens were
used as stimulator cells after irradiation with 30 Gy. MLR was
performed in triplicate wells. After 5 days incubation at
37.degree. C., 1 .mu.Ci [.sup.3H]-thymidine was added to each cup.
After a further 16 hours incubation, cells were harvested and
counted in a .beta.-counter. Inhibition of proliferation by a
compound (drug) described in some of the previous examples was
counted using the formula: Percent .times. .times. .times.
inhibition = ( cpm + drugs ) - cpm .times. .times. Cult . .times.
Med ( cpm - drugs ) - cpm .times. .times. Cult . .times. Med 100
##EQU2## wherein cpm is the thymidine count per minute. TNF-Alpha
and IL-1 Beta Assays
[0256] Peripheral blood mononuclear cells (herein referred as
PBMC), in response to stimulation by lipopolysaccharide (LPS), a
gram-negative bacterial endotoxin, produce various chemokines, in
particular human TNF-alpha and II-1 beta. The inhibition of the
activation of PBMC can be measured by the level of suppression of
the production of TNF-alpha or IL-1 beta by PBMC in response to
stimulation by LPS.
[0257] Such inhibition measurement was performed as follows: PBMC
were isolated from heparinized peripheral blood (Buffy coat) by
density gradient centrifugation. LPS is then added to the PMBC
suspension in complete medium (10.sup.6 cells/ml) at a final
concentration of 1 .mu.g/ml. The compound to be tested was added at
different dilution levels, and the cells were incubated at
37.degree. C. for 72 hours. The supernatants were collected, and
TNF-alpha or II-1 beta concentrations were measured with
respectively an anti-TNF antibody or an anti-IL-1 beta antibody in
a sandwich ELISA.
[0258] The percent inhibition was calculated as: %
inhibition=(pg/ml in sample-pg/ml min.)/(pg/ml max.-pg/ml min.)-1
wherein: min.: pg/ml in culture medium without test compound, and
[0259] max.: pg/ml in culture medium+LPS without test compound.
[0260] Table 2 below shows the IC.sub.50 values (expressed in
.mu.M) of the tested compounds in the MLR test and in the TNF and
IL-1 assays (ND: not determined) TABLE-US-00002 TABLE 2 Ex.
n.degree. R.sub.3 R.sub.4 MLR TNF IL-1 6 phenyl chloro 4.2 3.0 0.4
14 phenyl chloro 4.1 0.5 0.35 15 p-methoxyphenyl chloro 3.0 0.8 0.4
16 p-toluyl chloro 3.6 1.1 0.5 17 phenyl chloro 3.0 0.8 0.2 18
phenyl chloro 4.4 4.5 ND 19 m,p-dimethoxyphenyl chloro 5.1 1.1 ND
20 p-chlorophenyl chloro 3.9 3.8 0.4
EXAMPLE 27
Preparation of 3-methyl-6-aminouracil and 3-allyl-6-aminouracil
[0261] A suspension of 6-aminouracil (5 g, 39.3 mmole) in
1,1,1,3,3-hexamethyldisilazane (25 ml, 118 mmole) with a catalytic
amount of ammonium sulfate (20 mg) was refluxed for 2 hours until a
clear solution was obtained. Methyl iodide or allyl iodide (1.5
equivalent) was added and refluxed. Reaction was quenched with a
saturated sodium bicarbonate solution and the precipate formed was
filtered off, yielding the pure 3-methyl-6-amino-uracil with a 98%
yield, or the pure 3-allyl-6-amino-uracil with a 83% yield.
EXAMPLE 28
Preparation of 5,6-diamino-1-substituted uracils and
5,6-diamino-3-substituted uracils
[0262] 5,6-diamino-1-substituted uracils and
5,6-diamino-3-substituted uracils were prepared according to the
following general method described herein by reference to FIG. 8.
The monosubstituted urea (wherein R.sub.2 may be for instance
selected from the group consisting of alkyl, cycloalkyl, alkyl,
heterocycloalkyl, alkylaryl, arylalkyl and heterocycloalkyl) is
reacted with cyanoacetic acid, followed by refluxing in an aqueous
alkaline solution to yield a 6-amino-uracil derivative. In step
(b), a nitroso group is introduced at position 5 of the uracil ring
by reaction with sodium nitrite under aqueous acidic conditions. In
step (c), the reduction of the nitrosogroup is effected either
catalytically (e.g. Pt/H2) or chemically (e.g. sodium dithionite in
water or ammonium sulfide in water), yielding the
5,6-diamino-uracil derivative. The 1-substituted-6-aminouracil is
reacted in step (d) with the reactant R.sub.1X (wherein R.sub.1 may
be for instance selected from the group consisting of alkyl,
cycloalkyl and allyl, and X may be selected from the group
consisting of chloro, bromo and iodo) in the presence of a base
(such as potassium,carbonate) and a polar aprotic solvent.
[0263] More specifically the following procedure was performed: the
monosubstituted urea (40 mmole) and cyanoacetic acid (40 mmole)
were dissolved in 5 ml acetic anhydride, and the resulting mixture
was stirred at 60.degree. C. for 3 hours. After cooling to room
temperature, 30 ml ice water was added. The suspension was filtered
and the precipitate was washed with water to yield a crude product.
Recrystallization from methanol quantitatively yielded the pure
intermediate as a white solid. This intermediate (40 mmole) was
dissolved in an aqueous NaOH solution (33%, 50 ml) and stirred at
80.degree. C. for 10 minutes. After cooling to 40.degree. C., the
mixture was neutralized with acetic acid to pH 7, diluted with
water (50 ml) and stirred for another 30 minutes. The precipitate
was collected by filtration to yield the 6-amino-uracil derivative
as a white powder.
[0264] In the following step, the 6-amino-uracil derivative (20
mmole) was suspended in water (60 ml), then NaNO.sub.2 (24 mmole)
and acetic acid (1 ml) were added dropwise. The resulting mixture
was stirred at room temperature for 1 hour. A pink precipitate was
formed, which was filtered off and washed with water to yield the
corresponding 5-nitroso-6-amino-uracil derivative.
[0265] Then, Na.sub.2S.sub.2O.sub.4 (45 mmole) was added
portionwise to a suspension of the 5-nitroso-6-amino-uracil
derivative (15 mmole) in water (50 ml). The resulting mixture was
stirred at room temperature for 1 hour. The precipitate was
collected and washed with water to yield the corresponding
5,6-diamino-uracil as a grey powder.
[0266] The following compounds were synthesized in this way:
[0267] 5,6-diamino-1-methyluracil was obtained in a 90% yield from
commercially available 1-methyl6-amino-uracil;
[0268] 5,6-diamino-3-methyluracil was obtained in a 88% yield from
the 3-methyl-6-amino-uracil of example 27;
[0269] 5,6diamino-1-phenyluracil was obtained from phenylurea in a
62% yield;
[0270] 5,6-diamino-1-ethyluracil was obtained from ethylurea in a
47% yield;
[0271] 5,6-diamino-1-butyluracil was obtained from n-butylurea in a
45% yield;
[0272] 5,6-diamino-1-benzyluracil was obtained from benzylurea in a
52% yield;
[0273] 5,6-diamino-3-allyluracil was obtained in a 58% yield from
the 3-allyl-6-aminouracil of example 27.
EXAMPLES 29 TO 39
Preparation of 1,3,6-trisubstituted-7-hydroxy-lumazines
[0274] The 1,3,6-trisubstituted-7-hydroxy-lumazines (otherwise
named 1,3,6-trisubstituted-2,4,7(1H, 3H, 8H)-pteridinetriones) of
examples 29 to 39 were prepared from the relevant
5,6-diamino-1-substituted uracils and 5,6-diamino-3-substituted
uracils of example 28 in accordance with the general method
described herein with reference to FIG. 9. In step (a) of FIG. 9,
an uracil bearing a substituent R.sub.1 and/or R.sub.2 wherein
R.sub.1 and R.sub.2 may for instance be selected from the group
consisting of hydrogen, alkyl, cycloalkyl, arylalkyl and
heterocyclic is reacted in a polar protic solvent with a reactant
selected from the group consisting of alkyl arylglyoxylates, alkyl
alkylglyoxylates, alkyl heterocyclic glyoxylates, alkyl
arylalkylglyoxylates and alkyl heterocyclic alkylglyoxylates.
[0275] More specifically the following procedure was performed: to
a suspension of the appropriate 5,6-diamino-1,3-substituted uracil
made in example 28 (10 mmole) in water (200 ml), the appropriate
alkyl arylglyoxylate, alkyl alkylglyoxylate, alkyl heterocyclic
glyoxylate, alkyl arylalkylglyoxylate or alkyl heterocyclic
alkylglyoxylate (12.5 mmole) was added. The resulting mixture was
heated under reflux for 40 minutes. After cooling to room
temperature, the precipitate was collected. The crude product was
recrystallized from a mixture (1/1) of methanol and water to yield
the desired 1,3,6-trisubstituted-7-hydroxy-lumazine as yellowish
crystals or powder. The following table 3 provides the meaning of
substituents R.sub.1, R.sub.2 and R.sub.3 for each of the 7-hydroxy
lumazines of examples 29 to 39.
[0276] The synthesis of each individual compound is now briefly
described by indicating the starting uracil and glyoxylate, the
yield and physical form, and characterized by nuclear magnetic
resonance data (200 MHz, in DMSO-d6).
[0277] 6-(4-methoxyphenyl)-1-methyl-7-hydroxy-lumazine (example 29)
was obtained from 5,6-diamino-1-methyluracil and ethyl
4-methoxybenzoylformate in 87% yield as yellow crystals. .sup.1H
NMR (ppm): 11.65 (s, 1H), 8.07 (d, J=8.8 Hz, 2H), 7.04 (d, J=8.8
Hz, 2H), 3.82 (s, 3H), 3.43 (s, 3H).
[0278] 6-(3,4dimethoxyphenyl)-1-methyl-7-hydroxy-lumazine (example
30) was obtained from 5,6-diamino-1-methyluracil and
ethyl-3,4-dimethoxybenzoylformate in 88% yield as yellowish powder.
.sup.1H NMR (ppm): 12.07 (s, 1H), 7.33 (d, 1H), 7.30 (s, 1H), 7.11
(d, 1H), 3.83 (s, 3H), 3.80 (s, 3.44 (s, 3H).
[0279] 6-(4-fluorophenyl)-1-methyl-7-hydroxy-lumazine (example 31)
was obtained from 5,6-diamino-1-methyluracil and
ethyl-4-fluorobenzoylformate in 60% yield as a yellow powder.
.sup.1H NMR (ppm): 11.68 (s, 1H), 8.13 (m, 2H), 7.32 (t, 8.8Hz,
2H), 3.43 (s, 3H).
[0280] 6-(3-methoxyphenyl)-1-methyl-7-hydroxy-lumazine (example 32)
was obtained from 5,6-diamino-1-methyluracil and
ethyl-3-methoxybenzoylformate in 84% yield as a yellowish powder.
.sup.1H NMR (ppm): 11.68 (s, 1H), 7.65 (m, 1H), 7.40 (t, 1H), 7.02
(dd, 1H), 3.80 (s, 3H), 3.43 (s, 3H).
[0281] 6-(2,6-dimethoxyphenyl)-1-methyl-7-hydroxy-lumazine (example
33) was obtained from 5,6-diamino-1-methyluracil and
ethyl-3,4-dimethoxybenzoylformate in 92% yield as a yellow powder.
.sup.1H NMR (ppm): 13.2 (br., 1H), 11.64 (s, 1H), 7.05 (m, 3H),
7.30 (s, 1H), 3.73 (s, 3H), 3.68 (s, 1H), 3.44 (s, 3H).
[0282] 6-(2-chlorophenyl)-1-methyl-7-hydroxy-lumazine (example 34)
was obtained from 5,6-diamino-1-methyluracil and
ethyl-2-chlorobenzoylformate in 92% yield as a white powder.
.sup.1H NMR (ppm): 11.70 (s, 1H), 7.46 (m, 3H), 7.33 (m, 1H), 3.45
(s, 3H).
[0283] 6-(3-chlorophenyl)-1-methyl-7-hydroxy-lumazine (example 35)
was obtained from 5,6-diamino-1-methyluracil and
ethyl-3-chlorobenzoylformate in 56% yield as a yellow powder.
.sup.1H NMR (ppm): 11.74 (s, 1H), 8.11 (m, 2H), 7.52 (m, 2H), 3.44
(s, 1H).
[0284] 6-(4-cyanophenyl)-1-methyl-7-hydroxy-lumazine (example 36)
was obtained from 5,6-diamino-1-methyluracil and ethyl
4-cyanobenzoylformate in 88% yield as yellow crystals. .sup.1H NMR
(ppm): 11.75 (s, 1H), 8.28 (d, 2H), 7.95 (d, 2H), 3.44 (s, 3H).
[0285] 1-methyl-6-(4-methylphenyl)-7-hydroxy-lumazine (example 37)
was obtained from 5,6-diamino-1-methyluracil and ethyl
4-methylbenzoylformate in 74% yield as yellowish crystals. .sup.1H
NMR (ppm): 11.66 (s, 1H), 7.98 (d, 2H), 7.29 (d, 2H), 3.43 (s, 3H),
2.36 (s, 3H).
[0286] 1-benzyl-6-phenyl-7-hydroxy-lumazine (example 38) was
obtained from 1-benzyl-5,6-diamino-uracil and ethyl benzoylformate
in 90% yield as yellowish crystals.
[0287] 6-(4-chlorophenyl)-1-methyl-7-hydroxy-lumazine (example 39)
was obtained from 5,6-diamino-1-methyluracil and ethyl
4-chlorobenzoylformate in 40% as a white powder. TABLE-US-00003
TABLE 3 Example R.sub.2 R.sub.1 R.sub.3 29 methyl H 4-methoxyphenyl
30 methyl H 3,4-dimethoxyphenyl 31 methyl H 4-fluorophenyl 32
methyl H 3-methoxyphenyl 33 methyl H 2,6-dimethoxyphenyl 34 methyl
H 2-chlorophenyl 35 methyl H 3-chlorophenyl 36 methyl H
4-cyanophenyl 37 methyl H 4-methylphenyl 38 benzyl H phenyl 39
methyl H 4-chlorophenyl (end of Table 3)
EXAMPLES 40 TO 54
Preparation of 1,3,6-trisubstituted-7-halo-lumazines
[0288] The 1,3,6-trisubstituted-7-halo-lumazines of examples 40 to
54 were prepared from the corresponding
1,3,6-trisubstituted-7-hydroxy-lumazines such as those examples 29
to 39 in accordance with the method described herein with reference
to step (b) of FIG. 9. The relevant
1,3,6-trisubstituted-7-hydroxy-lumazine (5 mmole) was added to a
suspension of 0.5 g NaX (wherein X is chloro or bromo) in 20 ml
POX.sub.3 (wherein X is chloro or bromo). The resulting mixture was
heated at 90.degree. C. until the starting material completely
disappeared. The reaction mixture was concentrated under reduced
pressure to a syrup and then 30 g ice was added. After 30 minutes
stirring at room temperature, the precipitate was collected, washed
with water, dried naturally to yield a crude product.
Chromatography on silica gel (using a mixture MeOH/CH.sub.2Cl.sub.2
in a ratio ranging from 1/100 to 1/20) yielded the desired
7-halo-1,3,6-trisubstituted lumazine. Crystals could be obtained by
recrystallization from MeOH.
[0289] The synthesis of each individual compound is now briefly
described by indicating the starting compound, the yield, purity
(determined by high performance liquid chrmoatography) and physical
form, and characterized by nuclear magnetic resonance data (200
MHz, in DMSO-d6).
[0290] 7-chloro-6-(4-methoxyphenyl)-1-methyl-lumazine (example 40)
was obtained in 40% yield from the compound of example 29 as
yellowish crystals. Purity: 93.1%. .sup.1H NMR (ppm): 12.07 (s,
1H), 7.73 (d, 2H), 7.10 (d, 2H), 3.85 (s, 3H), 3.45 (s, 3H).
[0291] 7-chloro-6-(3,4-dimethoxyphenyl)-1-methyl-lumazine (example
41) was obtained from the compound of example 30 in 62% yield as
white crystals. Purity: 98.8%. .sup.1H NMR (ppm): 12.07 (s, 1H),
7.35 (d, 1H), 7.30 (s, 1H), 7.12 (d, H), 3.83 (s, 3H), 3.80 (s,
3H), 3.44 (s, 3H).
[0292] 7-chloro-6-(4-fluorophenyl)-1-methyl-lumazine (example 42)
was obtained from the compound of example 31 in 48% yield as
yellowish crystals. Purity (HPLC): 95.2%. .sup.1H NMR (ppm): 12.12
(s, 1H), 7.81 (m, 2H), 7.40 (m, 2H), 3.45 (s, 3H).
[0293] 7-chloro-6-(3-methoxyphenyl)-1-methyl-lumazine (example 43)
was obtained from the compound of example 32 in 64% yield as white
crystals. Purity: 94.1%; .sup.1H NMR (ppm): 12.11 (s, 1H), 7.48 (t,
1H), 7.26 (m, 2H), 7.11 (m, 1H), 3.82 (s, 3H), 3.45 (s, 3H).
[0294] 7-chloro-(2,6-dimethoxyphenyl)-1-methyl-lumazine (example
44) was obtained from the compound of example 33 in 75% yield as
white crystals. Purity: 98.3%. .sup.1H NMR (ppm): 12.06 (s, 1H),
7.10 (m, 2H), 6.88 (m, 1H), 3.75 (s, 3H), 3.72 (s, 3H), 3.44 (s,
3H).
[0295] 7-chloro-(2-chlorophenyl)-1-methyl-lumazine (example 45) was
obtained from the compound of example 34 in 54% yield as yellowish
crystals. Purity: 98.4%. .sup.1H NMR (ppm): 12.13 (s, 1H), 7.60 (m,
4H), 3.46 (s, 3H).
[0296] 7-chloro-3-(chlorophenyl)-1-methyl-lumazine (example 46) was
obtained from the compound of example 35 in 12% yield as yellowish
crystals. Purity: 97.0%.H NMR (ppm): 12.15 (s, 1H), 7.70 (m, 2H),
7.60 (m, 2H), 3.46 (s, 3H).
[0297] 7-chloro-1-ethyl-6-phenyl-lumazine (example 47) was obtained
from 7-hydroxy-1-ethyl-6-phenyl-lumazine (itself available by
reacting 5,6-diamino-1-ethyluracil and ethyl benzoylformate) in 79%
yield as white crystals. Purity: 97.5%. .sup.1H NMR (ppm): 12.08
(s, 1H), 7.74 (m, 2H), 7.54 (m, 3H), 4.12 (q, 2H), 1.23 (t,
3H).
[0298] 7-chloro6-(4-cyanophenyl)-1-methyl-lumazine (example 48) was
obtained from the compound of example 36 in 77% yield as yellowish
crystals. Purity: 98.5%. .sup.1H NMR (ppm): 12.18 (s, 1H), 8.05 (d,
2H), 7.95 (d, 2H), 3.46 (s, 3H).
[0299] 7-bromo-1-methyl-6-phenyl-lumazine (example 49) was obtained
from 7-hydroxy-1-methyl-6-phenyl-lumazine (itself available by
reacting 5,6-diamino-1-methyluracil and ethyl benzoylformate) in
70% yield as yellowish crystals. Purity: 99.2%. .sup.1H NMR (ppm):
8.90 (br.s, 1H), 7.76 (m, 2H), 7.48 (m, 2H), 3.70 (s, 3H).
[0300] 7-bromo-6-(3,4-dimethoxyphenyl)-1-methyl-lumazine (example
50) was obtained from the compound of example 30 in 50% yield as
yellow crystals. Purity: 100%. .sup.1H NMR (ppm): 8.58 (br.s, 1H),
7.40 (dd, 1H), 7.30 (d, 1H), 6.97 (d, 1H), 3.96 (s, 3H), 3.95 (s,
3H), 3.69 (s, 3H).
[0301] 7-bromo-6-(4-methoxyphenyl)-1-methyl-lumazine (example 51)
was obtained from the compound of example 29 in 48% yield as yellow
crystals. Purity: 99.3%. .sup.1H NMR (ppm): 8.73 (br.s, 1H), 7.80
(m, 1H), 7.03 (m, 2H), 3.91 (s, 3H), 3.71 (s, 3H), 3.69 (s,
3H).
[0302] 7-chloro-1-methyl-6-(4-methylphenyl)-lumazine (example 52)
was obtained from the compound of example 37 in 58% yield as white
crystals. Purity: 97.7%. .sup.1H NMR (ppm): 12.19 (s, 1H), 7.75 (d,
2H), 7.46 (d, 2H), 3.55 (s, 3H), 2.50 (s, 3H).
[0303] 7-chloro-1-benzyl-6-phenyl-lumazine (example 53) was
obtained from the compound of example 38 in 48% yield as yellowish
crystals. Purity: 97.4%. .sup.1NMR (ppm): 12.20 (s, 1H), 7.74 (m,
2H), 7.55 (m, 3H), 7.35 (m, 5H), 5.28 (s, 2H), 3.45 (s, 3H).
[0304] 7-chloro-6-(4-chlorophenyl)-1-methyl-lumazine (example 54)
was obtained from the compound of example 39 in 71% yield as
yellowish crystals. Purity: 99.7%. .sup.1H NMR (ppm): 12.13 (s,
1H), 7.77 (d, 2H), 7.64 (d, 2H), 3.45 (s, 3H).
[0305] The following table 4 summarizes the meanings of
substituents R.sub.1, R.sub.2, R.sub.3 and R.sub.4 for each of the
1,6-disubstituted 7-halo-lumazines of examples 40 to 54.
TABLE-US-00004 TABLE 4 Example R.sub.2 R.sub.1 R.sub.3 R4 40 methyl
H 4-methoxyphenyl Cl 41 methyl H 3,4-dimethoxyphenyl Cl 42 methyl H
4-fluorophenyl Cl 43 methyl H 3-methoxyphenyl Cl 44 methyl H
2,6-dimethoxyphenyl Cl 45 methyl H 2-chlorophenyl Cl 46 methyl H
3-chlorophenyl Cl 47 ethyl H phenyl Cl 48 methyl H 4-cyanophenyl Cl
49 methyl H phenyl Br 50 methyl H 3,4-dimethoxyphenyl Br 51 methyl
H 4-methoxyphenyl Br 52 methyl H 4-methylphenyl Cl 53 benzyl H
phenyl Cl 54 methyl H 4-chlorophenyl Cl (end of table 4)
EXAMPLES 55 TO 82
Preparation of 1.3,6-trisubstituted-7-halo-lumazines
[0306] The 7-halo-1,3,6-trisubstituted lumazines of examples 55 to
82 were prepared from the 7-halo-1,6-disubstituted lumazines of
examples 40 to 54 in accordance with the method described in FIG.
10 for introducing a further substituent R.sub.1 or R.sub.2. The
starting lumazine is reacted in step (a) with a reactant R.sub.2X
or in step (b) with a reactant R.sub.1X, wherein R.sub.1 and
R.sub.2 may each be for instance selected from the group consisting
of alkyl, cycloalkyl, alkyl carboxylic acid esters, thioesters and
amides, thiocarboxylic acid esters, thioesters and amides,
heterocycloalkyl, and wherein X may be for instance selected from
the group consisting of chloro, bromo, iodo, tosylate and mesylate,
in the presence of a base (such as, but not limited to, potassium
carbonate or sodium hydride) and a polar aprotic solvent.
[0307] More specifically, the following procedure was performed:
0.38 mmole of an alkyl halide (in which the halide is bromo or
iodo) was added to a mixture of a 7-halo-1,6-disubstituted
lumazine--step (a)--or a 7-halo-3,6-disubstituted lumazine--step
(b)--(0.30 mmole) and K.sub.2CO.sub.3 (62 mg, 0.45 mmole) in 4 ml
DMF. The resulting mixture was stirred at room temperature for 4
hours. The mixture was diluted with 50 ml CH.sub.2Cl.sub.2 and
washed with 40 ml brine. After drying over MgSO.sub.4, filtration
and concentration, the residue was purified by silica gel
chromatography to yield the desired 7-halo-1,3,6-trisubstituted
lumazine noted as 4 in FIG. 10.
[0308] The synthesis of each individual compound is now briefly
described by indicating the starting compound, the yield, purity
(determined by high performance liquid chromatography) and physical
form, and characterized by nuclear magnetic resonance (200 MHz, in
CDCl.sub.3).
[0309] 7-chloro-3-(ethyl butyrate)-1-methyl-6-phenyl-lumazine
(example 55) was obtained from 7-chloro-1-methyl-6-phenyl-lumazine
(which may itself be obtained from
7-hydroxy-1-methyl-6-phenyl-lumazine) in 79% yield as white
crystals. Purity: 99.9%. .sup.1H NMR (ppm): 7.82 (m, 2H), 7.50 (m,
3H), 4.21 (t, 2H), 4.11 (q, 2H), 3.72 (s, 3H), 2.43 (t, 2H), 2.07
(m, 2H), 1.24 (t, 3H).
[0310] 7-chloro-1-(ethyl butyrate)-3-methyl-6-phenyl-lumazine
(example 56) was obtained from 7-chloro-3-methyl-6-phenyl-lumazine
(which may itself be obtained from
7-hydroxy-3-methyl-6-phenyl-lumazine) in 62% yield as white
crystals. Purity: 98.4%. .sup.1H NMR (ppm): 7.82 (m, 2H), 7.50 (m,
3H), 4.41 (t, 2H), 4.12 (q, 2H), 3.55 (s, 3H), 2.46 (t, 2H), 2.14
(m, 2H), 1.25 (t, 3H).
[0311] 7-chloro-3-(2-hydroxyethyl)-1-methyl-6-phenyl-lumazine
(example 57) was obtained from 7-chloro-1-methyl-6-phenyl-lumazine
in 65% yield as white crystals. Purity: 98.8%. .sup.1H NMR (ppm):
7.82 (m, 2H), 7.50 (m, 3H), 4.41 (t, 2H), 3.97 (m, 2H), 3.74 (s,
3H).
[0312] 7-chloro-3-(2-phenylethyl)-1-methyl-6-phenyl-lumazine
(example 58) was obtained from 7-chloro-1-methyl-6-phenyl-lumazine
in 65% yield as white crystals. Purity: 98.6%. .sup.1H NMR (ppm):
7.82 (m, 2H), 7.50 (m, 3H), 7.32 (m, 5H), 4.35 (m, 2H), 3.72 (s,
3H), 3.01 (m, 2H).
[0313] 7-chloro-3-benzyl-1-methyl-6-phenyl-lumazine (example 59)
was obtained from 7chloro-1-methyl-6-phenyl-lumazine in 64% yield
as white crystals. Purity: 99.3%. .sup.1H NMR (ppm): 7.82 (m, 2H),
7.58 (m, 2H), 7.50 (m, 3H), 7.28 (m, 3H), 5.32 (s, 2H), 3.71 (s,
3H).
[0314] 7-chloro-1-(2-phenylethyl)-3-methyl-6-phenyl-lumazine
(example 60) was obtained from 7-chloro-3-methyl-6-phenyl-lumazine
in 34% yield as white crystals. Purity: 97.6%. .sup.1H NMR (ppm):
7.80 (m, 2H), 7.50 (m, 3H), 7.30 (m, 5H), 4.55 (m, 2H), 3.56 (s,
3H), 3.07 (m, 2H).
[0315] 1-benzyl-7-chloro-3-methyl-6-phenyl-lumazine (example 61)
was obtained from 7-chloro-3-methyl-6-phenyl-lumazine in 46% yield
as white crystals. Purity: 98.7%. .sup.1H NMR (ppm): 7.82 (m, 2H),
7.58 (m, 2H), 7.50 (m, 3H), 7.32 (m, 3H), 5.51 (s, 2H), 3.56 (s,
3H).
[0316] 7-chloro-1-(2-hydroxyethyl)-3-methyl-6-phenyl-lumazine
(example 62) was obtained from 7-chloro-3-methyl-6-phenyl-lumazine
in 42% as white crystals. Purity: 97.5%. .sup.1H NMR (ppm): 7.82
(m, 2H), 7.50 (m, 3H), 4.59 (t, 2H), 4.04 (t, 2H), 3.56 (s,
3H).
[0317] 7-chloro-6-(3,4-dimethoxyphenyl)-3-(ethyl
butyrate)-1-methyl-lumazine (example 63) was obtained from the
compound of example 41 and ethyl 4-bromobutyrate in 56% yield as a
yellow powder. Purity: 93.7%. .sup.1H NMR (ppm): 7.48 (dd, 1H),
7.38 (d, 1H), 6.97 (d, 1H), 4.22 (t, 2H), 4.12 (q, 2H), 3.96 (s,
6H), 3.72 (s, 3H), 2.43 (t, 2H), 2.08 (m, 2H), 1.24 (t, 3H).
[0318]
7-chloro-6-(3,4-dimethoxyphenyl)-3-(2-hydroxyethyl)-1-methyl-lumaz-
ine (example 64) was obtained from the compound of example 41 and
2-bromoethanol in 70% yield as a white yellow powder. Purity:
98.4%. .sup.1H NMR (ppm): 7.47 (dd, 1H), 7.37 (d, 1H), 6.97 (d,
1H), 4.41 (m, 2H), 3.98 (m, 2H), 3.96 (s, 6H), 3.74 (s, 3H).
[0319] 7-chloro-1,6-diphenyl-3-(ethyl butyrate)-lumazine (example
65) was obtained from 7-chloro-1,6-diphenyl-lumazine and ethyl
4-bromobutyrate in 67% yield as white crystals. Purity: 99.8%.
.sup.1H NMR (ppm): 7.79 (m, 2H), 7.57 (m, 3H), 7.49 (m, 3H), 7.32
(m, 2H), 4.25 (t, 2H), 4.10 (q, 2H), 2.45 (t, 2H), 2.12 (m, 2H),
1.26 (t, 3H).
[0320] 7-chloro-1,6-diphenyl-3-(2-hydroxyethyl)-lumazine (example
66) was obtained from 7chloro-1,6diphenyl-lumazine and
2-bromoethanol in 78% yield as white crystals. Purity: 91%. .sup.1H
NMR (ppm): 7.79 (m, 2H), 7.57 (m, 3H), 7.49 (m, 3H), 7.32 (m, 2H),
4.35 (t, 2H), 3.42 (t, 2H).
[0321] 3-butyronitrile-7-chloro-1-methyl-6-phenyl-lumazine (example
67) was obtained from 7-chloro-1-methyl-6-phenyl-lumazine and
4-bromobutyronitrile in 89% yield as white crystals. Purity: 99.3%.
.sup.1H NMR (ppm): 7.82 (m, 2H), 7.49 (m, 3H), 4.31 (t, 2H), 3.73
(s, 3H), 2.49 (t, 2H), 2.15 (m, 2H).
[0322] 7-chloro-1-ethyl-3-(ethyl butyrate)-6-phenyl-lumazine
(example 68) was obtained from the compound of example 47 and ethyl
4-bromobutyrate in 95% yield as white crystals. Purity: 99.2%.
.sup.1H NMR (ppm): 7.82 (m, 2H), 7.50 (m, 3H), 4.38 (q, 2H), 4.21
(t, 2H), 4.10 (q, 2H), 2.43 (t, 2H), 2.07 (m, 2H), 1.35 (t, 3H),
1.24 (t, 3H).
[0323] 3-acetamido-7-chloro-1-methyl-6-phenyl-lumazine (example 69)
was obtained from 7-chloro-1-methyl-6-phenyl-lumazine and
2-bromoacetamide in 95% yield as white crystals. Purity: 98.8%.
.sup.1H NMR (ppm): 7.82 (m, 2H), 7.50 (m, 3H), 4.82 (s, 2H), 3.74
(s, 3H).
[0324] 7-bromo-3-(ethyl butyrate)-1-methyl-6-phenyl-lumazine
(example 70) was obtained from the compound of example 49 and ethyl
4-bromobutyrate in 69% yield as yellowish crystals. Purity: 98.2%.
.sup.1H NMR (ppm): 7.74 (m, 2H), 7.49 (m, 3H), 4.21 (t, 2H), 4.10
(q, 2H), 3.73 (s, 3H), 2.43 (t, 2H), 2.07 (m, 2H), 1.24 (t,
3H).
[0325] 7-chloro-3-(ethyl acetate)-1-methyl-6-phenyl-lumazine
(example 71) was obtained from 7-chloro-1-methyl-6-phenyl-lumazine
and ethyl 2-bromoacetate in 97% yield as a white powder. Purity:
99.2%. .sup.1H NMR (ppm): 7.82 (m, 2H), 7.50 (m, 3H), 4.88 (s, 3H),
4.25 (q, 2H), 3.74 (s, 3H), 1.31 (t, 3H).
[0326] 7-chloro-3-(ethyl pentanoate)-1-methyl-6-phenyl-lumazine
(example 72) was obtained from 7-chloro-1-methyl-6-phenyl-lumazine
and ethyl 5-bromopentanoate in 65% yield as a white powder. Purity:
99.7%. .sup.1H NMR (ppm): 7.82 (m, 2H), 7.50 (m, 3H), 4.13 (m, 4H),
3.72 (s, 3H), 2.37 (t, 2H), 1.75 (m, 4H), 1.25 (t, 3H).
[0327] 7-chloro-3-(ethyl
butyrate)-1-methyl-6-(4-methoxyphenyl)-lumazine (example 73) was
obtained from the compound of example 40 and ethyl-4-bromobutyrate
in 55% yield as a yellow solid. Purity: 97.0%. .sup.1H NMR (ppm):
7.82 (d, 2H), 7.01 (d, 2H), 4.21 (t, 2H), 4.11 (q, 2H), 3.88 (s,
3H), 3.71 (s, 3H), 2.43 (t, 2H), 2.08 (m, 2H), 1.24 (t, 3H).
[0328] 7-chloro-1-methyl-3-(2-morpholinoethyl)-6-phenyl-lumazine
(example 74) was obtained from 7-chloro-1-methyl-6-phenyl-lumazine
and 4-(2-iodoethyl)morpholine in 88% yield as white crystals.
Purity: 91.2%. .sup.1H NMR (ppm): 7.82 (m, 2H), 7.50 (m, 3H), 4.31
(t, 2H), 3.73 (s, 3H), 3.67 (m, 4H), 2.60 (m, 4H).
[0329] 3-(2-butyloxycarbonyl-aminoethyl)-7-chloro-1-methyl
-6-phenyl-lumazine (example 75) was obtained from
7-chloro-1-methyl-6-phenyl-lumazine and
2-(butyloxycarbonylamino)ethyl bromide in 61% as white crystals.
Purity: 99.5%. .sup.1H NMR (ppm): 7.82 (m, 2H), 7.50 (m, 3H), 4.90
(br., 1H), 4.32 (t, 2H), 3.73 (s, 3H), 3.52 (m, 2H), 1.32 (s,
9H).
[0330] 7-chloro-1,3-dimethyl-6-(4-methoxyphenyl)-lumazine (example
76) was obtained from the compound of example 40 and iodomethane in
57% yield as a yellow solid. Purity: 99.9%. .sup.1H NMR (ppm): 7.82
(d, 2H), 7.01 (d,2H),13.88 (s, 3H), 3.73 (s, 3H), 3.56 (s, 3H).
[0331] 7-chloro-1-methyl-6-phenyl-3-(2-piperidinoethyl)-lumazine
(example 77) was obtained from 7-chloro-1-methyl-6-phenyl-lumazine
and 1-(2-iodoethyl)piperidine in 54% yield as a yellow solid.
Purity: 91.7%. .sup.1H NMR (ppm): 7.82 (d, 2H), 7.01 (d, 2H), 4.38
(m, 2H), 3.66 (s, 3H), 2.90 (m, 4H), 1.60(m, 6H).
[0332] 7-chloro-1-methyl-6-phenyl-3-(2-pyrrolidinoethyl)-lumazine
(example 78) was obtained from 7-chloro-1-methyl-6-phenyl-lumazine
and 1-(2-iodoethyl)pyrrolidine in 66% yield as yellow solid.
Purity: 91.9%. .sup.1H NMR (ppm): 7.82 (d, 2H), 7.01 (d, 2H), 4.50
(m, 2H), 3.70 (s, 3H), 3.56 (m, 2H), 2.12 (m, 4H), 1.30 (m,
2H).
[0333]
7-bromo-6-(3,4-dimethoxyphenyl)-3-(ethylbutyrate)-1-methyl-lumazin-
e (example 79) was obtained from the compound of example 50 and
ethyl 4-bromobutyrate in 54% yield as a yellow solid. Purity:
99.7%; .sup.1H NMR (ppm): 7.42 (dd, 1H), 7.31 (d, 1H), 6.96 (d,
1H), 4.21 (t, 2H), 4.11 (q, 2H), 3.95 (s, 6H), 3.72 (s, 3H), 2.43
(t, 2H), 2.07 (m, 2H), 1.24 (t, 3H).
[0334]
7-bromo-3-(ethylbutyrate)-1-methyl-6-(4-methoxyphenyl)-lumazine
(example 80) was obtained from the compound of example 51 and ethyl
4-bromobutyrate in 55% yield as yellow solid. Purity: 99.4%.
.sup.1H NMR (ppm): 7.79 (d, 2H), 7.03 (d, 2H), 4.23 (t, 2H), 4.14
(q, 2H), 3.91 (s, 3H), 3.74 (s, 3H), 2.45 (t, 2H), 2.10 (m, 2H),
1.27 (t, 3H).
[0335] 7-bromo-1-methyl-3-(2-morpholinoethyl)-6-phenyl-lumazine
(example 81) was obtained from the compound of example 49 and
4-(2-iodoethyl)morpholine in 68% yield as a yellowish powder.
Purity: 95.5%. .sup.1H NMR (ppm): 7.77 (m, 2H), 7.50 (m, 3H), 4.31
(t, 2H), 3.74 (s, 3H), 3.69 (m, 4H), 2.65 (m, 4H).
[0336]
7-bromo-1-methyl-3-(2-morpholinoethyl)-6-(3,4dimethoxyphenyl)-luma-
zine (example 82) was obtained from the compound of example 50 and
4-(2-iodoethyl)morpholine in 76% yield as a yellow powder. Purity:
90.3%. .sup.1H NMR (ppm): 7.42 (dd, 1H), 7.32 (d, 1H), 6.98 (d,
1H), 4.32 (t, 2H), 3.96 (s, 6H), 3.73 (s, 3H), 3.72 (m, 4H), 2.70
(m, 4H).
[0337] The following table 5 summarizes the meanings of
substituents R.sub.1, R.sub.2, R.sub.3 and R.sub.4 for each of the
1,3,6-trisubstituted 7-halolumazines of examples 55 to 82.
TABLE-US-00005 TABLE 5 Example R.sub.2 R.sub.1 R.sub.3 R.sub.4 55
methyl ethyl butyrate phenyl Cl 56 ethyl butyrate Me phenyl Cl 57
methyl 2-hydroxyethyl phenyl Cl 58 methyl 2-phenylethyl phenyl Cl
59 methyl benzyl phenyl Cl 60 2-phenylethyl methyl phenyl Cl 61
benzyl methyl phenyl Cl 62 2-hydroxyethyl methyl phenyl Cl 63
methyl ethyl butyrate 3,4- Cl dimethoxyphenyl 64 methyl
2-hydroxyethyl 3,4- Cl dimethoxyphenyl 65 phenyl ethyl butyrate
phenyl Cl 66 phenyl 2-hydroxyethyl phenyl Cl 67 methyl
4-butyronitrile phenyl Cl 68 ethyl ethyl butyrate phenyl Cl 69
methyl 2-acetamido phenyl Cl 70 methyl ethyl butyrate phenyl Br 71
methyl ethyl acetate phenyl Cl 72 methyl ethyl pentanoate phenyl Cl
73 methyl ethyl butyrate 4-methoxyphenyl Cl 74 methyl
2-morpholinoethyl phenyl Cl 75 methyl 2-butyloxycarbonyl phenyl Cl
aminoethyl 76 methyl methyl 4-methoxyphenyl Cl 77 methyl
2-piperidinoethyl phenyl Cl 78 methyl 2-pyrrolidinoethyl phenyl Cl
79 methyl ethyl butyrate 3,4- Br dimethoxyphenyl 80 methyl ethyl
butyrate 4-methoxyphenyl Br 81 methyl 2-morpholinoethyl phenyl Br
82 methyl 2-morpholinoethyl 3,4- Br dimethoxyphenyl 83 methyl ethyl
butyrate 4-methoxyphenyl F 84 methyl Me 4-hydroxyphenyl Br
EXAMPLE 83
Preparation of 3-(ethyl
butyrate)-7-fluoro-1-methyl-6-(4-methoxyphenyl)-lumazine
[0338] CsF (1.2 mmole, 200 mg) and 18-crown-6 (0.15 mmole, 40 mg)
were added to a mixture of 4A' molecular sieves (1.0 g) in THF (4
ml). The resulting mixture was stirred at room temperature for 1
hour. Then the compound of example 73 (0.5 mmole) was added. The
mixture was stirred at room temperature for 3 hours, and then
filtered through a pad of Celite and rinsed with CH.sub.2Cl.sub.2.
After concentration under reduced pressure, the residue was
purified by chromatography on silica gel (2% acetone in
CH.sub.2Cl.sub.2) to yield the desired
3-(ethylbutyrate)-7-fluoro-1-methyl-6-(4-methoxyphenyl)-lumazine in
81% yield as a yellowish solid. .sup.1H NMR (ppm): 8.12 (m, 2H),
7.02 (m, 3H), 4.23 (t, 2H), 4.13 (q, 2H), 3.90 (s, 3H), 3.69 (s,
3H), 2.45 (t, 2H), 2.10 (m, 2H), 1.26 (t, 3H).
EXAMPLE 84
Preparation of
7-bromo-1,3-dimethyl-6-(4-hydroxyphenyl)-lumazine
[0339] Boron tribromide (1.2 mmole) was added to a solution of the
compound of example 76 (0.4 mmole) in 4 ml CH.sub.2Cl.sub.2. The
resulting mixture was stirred at room temperature for 24 hours.
Reaction was then quenched with water, and the mixture extracted
with CH.sub.2Cl.sub.2. After concentration under reduced pressure,
the residue was purified by chromatography on silica gel (1/30
acetone/CH.sub.2Cl.sub.2) to yield
7-bromo-1,3-dimethyl-6-(4-hydroxyphenyl)-lumazine in 90% yield as a
yellow solid. .sup.1H NMR (ppm): 7.63 (m, 2H), 6.96 (m, 2H), 6.21
(s, 1H), 3.77 (s, 3H), 3.60 (s, 3H). Interestingly, this procedure
achieves simultaneous introduction of a bromo substituent at
position 7 of the pteridine ring and conversion of a methoxyphenyl
to a hydroxyphenyl substituent at position 6 of the pteridine
ring.
EXAMPLES 85 TO 92
Preparation of 3-carboxy-7-halo-1,6-substituted lumazines
[0340] A 7-halo-1,6-disubstituted-3-(ethyl butyrate) lumazine is
converted into the corresponding
7-halo-1,6-disubstituted-3-(n-butyric acid) lumazine (noted 5 in
FIG. 11) in accordance with the method shown in FIG. 11. For
reasons of clarity, the synthesis is only shown for the ethyl ester
of n-butyric acid, although the synthetic procedure outlined
hereunder is similarly applicable to other esters of other
monocarboxylic acids. In step (a), the ethyl ester group of a
tetra-substituted lumazine is converted into the corresponding free
carboxylic acid by basic or acidic hydrolysis. Then in step (b),
the 7-halo-1,6-disubstituted-3-(carboxylic acid) lumazine may be
reacted with a nucleophile (e.g. an amine, alcohol, phenol, thiol
or thiophenol) having the general formula HR.sub.5, wherein R.sub.5
may be for instance selected from the group consisting of amino,
alkylamino, cycloalkylamino, arylamino, heterocyclic alkylamino,
alkoxy, aryloxy, alkylaryloxy, arylalkoxy, heterocyclic alkoxy,
thioalkyl, arylthio, arylalkylthio alkylarylthio and heterocyclic
alkylthio) in a protic solvent or aprotic solvent in the presence
of an acid catalyst. Suitable examples of such acid catalysts
include, but are not limited to, acetyl chloride and
trifluoroacetic anhydride. Alternatively, the monocarboxylic acid
can first be converted into the corresponding monocarboxylic acid
halide (e.g. by reaction with thionyl chloride or oxalyl chloride),
followed by reaction with the said nucleophile HR.sub.5.
[0341] More specifically, the following procedure was performed for
step (a): a solution of a 7-halo-1,6-disubstituted-3-(ethyl
butyrate) lumazine (0.5 mmole) in 10 ml dioxane and 10 ml 5% HCl
was stirred at room temperature for 24 hours. The solvents were
removed under reduced pressure to yield the corresponding
7-halo-1,6-disubstituted-3-n-butyric acid) lumazine which was
analyzed by high performance liquid chromatography (purity) and
nuclear magnetic resonance (200 MHz, in CDCl.sub.3).
[0342] 7-chloro-3-(n-butyric acid)-1-methyl-6-phenyl-lumazine
(example 85) is thus obtained from the compound of example 55 in
99% yield as white crystals. Purity: 97.0%. .sup.1H NMR (ppm): 7.80
(m, 2H), 7.49 (m, 3H), 4.23 (t, 2H), 3.72 (s, 3H), 2.47 (t, 2H),
2.08 (m, 2H).
[0343] 7-chloro-3-(n-butyric acid)-1-ethyl-6-phenyl-lumazine
(example 86) is thus obtained from the compound of example 68 in
99% yield as white crystals. Purity: 90%. .sup.1H NMR (ppm): 7.82
(m, 2H), 7.49 (m, 3H), 4.38 (q, 2H), 4.23 (t, 2H), 2.47 (t, 2H),
2.08 (m, 2H), 1.38 (t, 3H).
[0344] The following procedure was performed for making isopropyl
esters (noted 6 in FIG. 11) according to step (b): acetylchloride
(200 .mu.l) was added to a solution of the compound of example 85
(0.4 mmole) in isopropyl alcohol (10 ml). The resulting mixture was
stirred at room temperature for 24 hours and then neutralized with
NaHCO.sub.3 to pH 7. After filtration and concentration, the
residue was purified by chromatography on silica gel to obtain
7-chloro-3-(isopropyl 4-butyrate)-1-methyl-6-phenyl-lumazine
(example 87) in 92% yield as a white powder. Purity: 97.5%. .sup.1H
NMR (ppm): 7.80 (m, 2H), 7.49 (m, 3H), 4.99 (m, 1H), 4.21 (t, 2H),
3.72 (s, 3H), 2.40 (t, 2H), 2.06 (m, 2H), 1.22 (d, 6H).
[0345] Following the above procedures for steps (a) and (b),
7-bromo-6-(3,4-dimethoxyphenyl)-3-(isopropyl
butyrate)-1-methyl-lumazine (example 88) was obtained from the
compound of example 79 in 54% yield as a yellow powder. Purity:
92.6%. .sup.1H NMR (ppm): 7.42 (dd, 1H), 7.31 (d, 1H), 6.96 (d,
1H), 5.01 (m, 1H), 4.26 (t, 2H), 3.98 (s, 6H), 3.75 (s, 3H), 2.42
(t, 2H), 2.08 (m, 2H), 1.24 (d, 6H).
[0346] The following procedure was performed for making
7-chloro-3-(t-butyl butyrate)-1-methyl-6-phenyl-lumazine (example
89) according to step (b): tert-butanol (95 .mu.l) and
trifluoroacetic anhydride (56 .mu.l) were added to a solution of
the compound of example 85 (0.2 mmole) in CH.sub.2Cl.sub.2 (4 ml).
The mixture was stirred at room temperature for 4 hours. After
removing the solvents under reduced pressure, the residue was
purified by chromatography on silica gel to yield the desired
t-butyl ester (76%) as a white powder. Purity: 98.9%. .sup.1H NMR
(ppm): 7.80 (m, 2H), 7.49 (m, 3H), 4.19 (t, 2H), 3.72 (s, 3H), 2.40
(t, 2H), 2.02 (m, 2H), 1.42 (s, 9H).
[0347] The following procedure was performed for making carboxylic
amides (noted 6 in FIG. 11) according to step (b): a solution of
the compound of example 85 (0.2 mmole) in thionyl chloride (2 ml)
was refluxed for 1 hour. The excess thionyl chloride was removed
under reduced pressure. The residue was dissolved in
CH.sub.2Cl.sub.2 (4 ml) and cooled to -78.degree. C., then a
suitable amine (or a salt thereof) (0.3 mmole) was added and pH was
adjusted above 9 by means of triethylamine. The mixture was stirred
at -78.degree. C. for 20 minutes, warmed to -20.degree. C., then
quenched with 5% HCl and extracted with CH.sub.2Cl.sub.2. After
drying and filtration, the residue was purified by chromatography
on silica gel to yield the desired carboxylic amide. In this way,
3-(4-butyramido)-7-chloro-1-methyl6-phenyl-lumazine (example 90)
was obtained from ammonia (33% aqueous solution) in 63% yield as a
yellowish solid. Purity: 98.0%. .sup.1H NMR (ppm): 7.80 (m, 2H),
7.49 (m, 3H), 6.10 (br., 1H), 5.68 (br., 1H), 4.23 (t, 2H), 3.73
(s, 3H), 2.34 (t, 2H), 2.10 (m, 2H).
[0348] Similarly, 7-chloro-3-(N-methyl
4-butyramido)-1-methyl-6-phenyl-lumazine (example 91) was obtained
from methylamine hydrochloride in 62% yield as a yellowish solid.
Purity: 92.0%. .sup.1H NMR (ppm): 7.79 (m, 2H), 7.49 (m, 3H), 6.15
(br., 1H), 4.20 (t, 2H), 3.73 (s, 3H), 2.78 (d, 3H), 2.28 (t, 2H),
2.10 (m, 2H). 7-chloro-1-methyl-6-phenyl-3-(N-propyl
4-butyramido)-lumazine (example 92) was obtained from propylamine
in 51% yield as a white solid. Purity: 98.9%. .sup.1H NMR (ppm):
7.80 (m, 2H), 7.50 (m, 3H), 6.05 (br., 1H), 4.20 (t, 2H), 3.73 (s,
3H), 3.18 (q, 2H), 2.27 (t, 2H), 1.52 (m, 2H), 0.91 (t, 3H).
[0349] Table 6 summarizes the meanings of substituents R.sub.2,
R.sub.5, R.sub.3 and R.sub.4 for each of the
3carboxy-1,6-disubstituted-7-halo-lumazines of examples 85 to 92.
TABLE-US-00006 TABLE 6 Example R.sub.2 R.sub.5 R.sub.3 R.sub.4 85
methyl OH phenyl Cl 86 ethyl OH phenyl Cl 87 methyl
OCH(CH.sub.3).sub.2 phenyl Cl 88 methyl OCH(CH.sub.3).sub.2
3,4-dimethoxyphenyl Br 89 methyl OC(CH.sub.3).sub.3 phenyl Cl 90
methyl NH.sub.2 phenyl Cl 91 methyl NHCH.sub.3 phenyl Cl 92 methyl
NH-n-propyl phenyl Cl
EXAMPLE 93
In vitro Biological Evaluation of Polysubstituted Lumazines
[0350] Table 7 below shows the IC.sub.50 values (expressed in
.mu.M) of some of the compounds of examples 40 to 92 in the MLR
test and in the TNF-.alpha. and IL-1 assays performed under the
same experimental conditions as in example 26. TABLE-US-00007 TABLE
7 MLR TNF.sub..alpha. IL.sub.1.beta. Example IC.sub.50(.mu.M)
IC.sub.50(.mu.M) IC.sub.50(.mu.M) 40 3.3 0.8 0.6 41 5.1 1.7 0.7 42
4.8 4.7 0.7 43 4.9 3.7 2.0 44 4.9 5.9 0.5 45 3.9 2.3 0.3 46 5.7 4.7
0.9 48 4.3 2.5 0.4 50 4.0 1.0 0.6 51 3.6 4.6 5.7 52 3.5 1.1 0.5 53
4.4 4.5 1.2 54 3.9 3.8 0.5 55 4.9 0.5 0.6 56 4.9 0.5 0.6 57 5.0 0.4
0.7 58 5.6 5.5 0.5 59 5.0 5.3 0.5 60 3.5 5.8 4.1 61 3.7 0.7 0.5 62
2.5 0.6 0.6 63 5.7 0.6 0.9 64 5.7 0.7 0.5 65 3.8 0.8 0.5 67 4.8 0.7
0.6 68 6.8 3.1 0.6 69 4.5 5.8 3.4 70 5.0 5.5 3.5 71 2.8 3.4 0.6 72
3.5 3.4 0.6 73 4.9 0.6 0.5 74 4.2 2.6 0.7 75 5.1 3.5 0.7 76 4.5 0.6
0.6 77 3.4 3.7 0.7 78 4.4 7.4 0.8 79 4.7 0.7 0.2 80 5.6 2.1 6.4 81
4.7 1.7 5.4 82 4.6 10.0 5.7 83 4.5 4.9 6.4 84 4.1 0.9 3.9 85 5.0
6.1 3.9 86 4.6 8.4 2.5 87 5.4 6.1 2.8 88 4.7 0.7 7.5 89 4.2 5.6 2.1
90 3.9 4.9 1.6 92 3.4 5.4 1.1 Table 7 (end)
EXAMPLE 94
Model of Rheumatoid Arthritis
[0351] Collagen type II (hereinafter referred as CII) induced
experimental model of rheumatoid arthritis (hereinafter referred as
RA) in DBA mice is widely accepted as the most relevant and
predictive preclinical model for RA. In this model, DBA mice are
immunized with CII, the collagen type mainly present in the joint
structures, together with complete Freund Adjuvant in their tail. 2
to 3 weeks later, several of the immunized mice start to develop
arthritis in the four footpaths. In order to further worsen the
disease, mice are given a second CII boost at three weeks after the
first immunisation, this time however in a footpath. Because the
immune system is already immunised in these mice, this rapidly
provokes a severe swelling of the injected footpath (named Delayed
Type Hypersensitivity or DTH) which can be used as a measurement
for T-cell activation. Within a few days after the booster, almost
all untreated animals start developing symptoms of arthritis. RA
development is scored from 0 to 16 (16 being severe clinical
arthritis in all four footpaths). At the end of the study (3 weeks
after the CII boost) antibody formation was determined against CII
and histology performed on the footpaths.
[0352] The efficiency of the polysubstituted lumazine of example
41, administered in an amount of 20 mg/kg/day, started one day
before the CII boost) was explored in this CII model. All such
treated animals developed significanly less severe rheumatoid
arthritis (clinical scores ranging from 0 to 5) as compared to
untreated control mice (clinical scores from 6 to 12) and also
compared to mice treated with methotrexate (clinical scores ranging
from 2 to 7), the most effective compound for the treatment of
rheumatoid arthritis to date.
EXAMPLE 95
Model of Multiple Sclerosis
[0353] The polysubstituted lumazine of example 42 showed
significant protection in a murine multiple sclerosis standard
model (experimental allergic encephalomyelitis) such as described
by Kuschnaroff et al. in J. Neuroimmunol. (1999) 99:157-168.
Animals that were treated with this lumazine (administered in an
amount of 20 mg/kg/day) developed less severe multiple sclerosis
(clinical scores from 0 to 1) when compared to untreated control
animals (clinical scores ranging from 3 to 4). Animals treated with
mitoxantrone (the only drug approved for multiple sclerosis) gave
similar results as the polysubstituted lumazine of example 42.
However, mitoxantrone induces heart and kidney toxicity, whereas
our lumazine did not show any sign of toxicity.
* * * * *