Note: Descriptions are shown in the official language in which they were submitted.
CA 02495455 2005-02-15
WO 20041022057 PCTIEP20031008628
Aventis Pharma Deutschland GmbH DEAV2002/0064 Dr. TH
Description
Use of IKB-kinase inhibitors in pain therapy
The invention relates to the use of IKB-kinase inhibitors for treating pain.
Patent applications WO 01/00610, WO 01/30774 and WO 01/68648
describe compounds which are able to modulate NFKB.
NFKB is a heterodimeric transcription factor which is able to activate a large
number of genes which encode, inter alia, proinflammatory cytokines such
as IL-1, IL-2, TNFa or IL-6. NFKB is present in the cytosol of cells, where it
is complexed with its naturally occurring inhibitor I,cB. Stimulation of the
cells, for example by cytokines, leads to the IKB being phosphorylated and
subsequently broken down proteolytically. This proteolytic breakdown leads
to the activation of NFKB, which then migrates into the nucleus of the cell,
where it activates a large number of proinflammatory genes.
In diseases such as rheumatoid arthritis (in connection with inflammation),
osteoarthritis, asthma, cardiac infarction, Alzheimer's diseases or
atherosclerosis, NF,cB is activated to beyond the normal extent. The
inhibition of NFKB is also of value in the treatment of cancer since it is
used
in such treatment to augment the cytostatic therapy. It has been
demonstrated that pharmaceuticals such as glucocorticoids, salicylates or
gold salts, which are used in the therapy of rheumatism, inhibit the NFKB-
activating signal chain at various points or interfere directly with the
transcription of the genes.
The first step in said signal cascade is the breakdown of IKB. This
phospho.rylation is regulated by the specific IKB kinase.
Pharmaceuticals belonging to a large number of different substance groups
are employed in treating acute and chronic pain. Despite this, the therapy
of pain has still not been satisfactorily solved even today. This is due, in
particular, to the fact that the analgesics which are on the market do not
have a sufficiently powerful effect.
In an endeavor to obtain active compounds for treating pain, it has now
been found that it is possible to use I,~B-kinase inhibitors for this purpose.
CA 02495455 2005-02-15
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In particular, it has been possible to demonstrate, in the models employed,
a strength of effect which is clearly superior to that of classical
nonsteroidal
anti-inflammatory agents.
The invention relates, therefore, to the use of IKB-kinase inhibitors for
producing pharmaceuticals for treating pain.
The term "pain" is understood as meaning acute pains and chronic pains.
The following are examples of chronic pains:
chronic musculoskeletal diseases, such as back pains,
pains associated with menstrual bleeding,
pains associated with osteoarthritis or rheumatoid arthritis,
pains associated with intestinal inflammation,
pains associated with cardiac muscle inflammation,
pains associated with multiple sclerosis,
pains associated with neuritis,
pains associated with carcinomas and sarcomas,
pains associated with AIDS,
pains associated with chemotherapy,
amputation pain,
trigeminus neuralgia,
headaches, such as migraine cephalalgia, or
neuropathic pains, such as post-herpes zoster neuralgia.
The following are examples of acute pains:
pains following injuries,
post-operative pains,
pains in association with an acute attack of gout, or
acute pains following jaw-bone surgery interventions.
Examples of IKB-kinase inhibitors are indole derivatives or benzimidazole
derivatives as are described in the patent applications WO 01/00610 and
WO 01!30774.
The invention furthermore relates to the use of compounds of the formula I
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3
R°
Ra
R~~ I \ (I)
~N / Rz
w Rs
R~
and/or a stereoisomeric form of the compound of the formula I and/or a
physiologically tolerated salt of the compound of the formula I,
for producing pharmaceuticals for treating pains, where
E is N atom or the radical -C(R'9)-,
where R'9 is hydrogen atom or the radical R9,
one of the substituents R', R2, R3 and R4 is a radical of the formula II,
N~
\Di RB I Z (11)
Rs
in which D is -C(O)-, -S(O)- or -S(O)2-,
Ra is hydrogen atom or -(C~-C4)-alkyl,
R9 is 1. characteristic radical of an amino acid,
2. aryl, in which aryl is unsubstituted or substituted,
3. heteroaryl having from 5 to 14 ring members, in which
heteroaryl is unsubstituted or substituted,
4. heterocycle having from 5 to 12 ring members, in
which heterocycle is unsubstituted or substituted,
5. -(C~-C6)-alkyl, in which alkyl is straight-chain or
branched and is unsubstituted or substituted, once,
twice or three times, independently of each other, by
5.1 aryl, in which aryl is unsubstituted or
substituted,
5.2 heteroaryl having from 5 to 14 ring members, in
which heteroaryl is unsubstituted or substituted,
5.3 heterocycle having from 5 to 12 ring members,
in which heterocycle is unsubstituted or
substituted,
5.4 -O-R",
5.5 =O,
5.6 halogen,
5.7 -CN,
5.8 -CF3,
5.9 -S(O)X R", in which x is the integer zero, 1 or 2,
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4
5.10 -C(O)-O-R",
5.11 -C(O)-N(R")Z,
5.12 -C(O)-R",
5.13 -N(R")Z,
5.14 -(C3-C6)-cycloalkyl,
5.15 radical of the
formula
Rii
Ri 1
R11
or
5.1 6 radical of the formula R11
R" is a) hydrogen atom,
b) (C~-C6)-alkyl, in which alkyl is unsubstituted
or
substituted once, twice or three times
1. aryl, in which aryl is unsubstituted
or
substituted,
2. heteroaryl having from 5 to 14 ring
members,
3. heterocycle having from 5 to 12 ring
members,
4. halogen,
5. -N-(C~-C6)~-alkyl, in which n is the
integer zero,
1 or 2 and alkyl is unsubstituted or
substituted
once, twice or three times, independently
of
each other, by halogen or by -C(O)-OH,
6. -O-(C~-C6)-alkyl or
7. -C(O)-OH,
c) aryl, in which aryl is unsubstituted
or substituted,
d) heteroaryl having from 5 to 14 ring members,
or
e) heterocycle having from 5 to 12 ring
members, and,
in the case of (R")2, R" has, independently
of each
other, the
meanings of
a) to e),
Z is 1. aryl, in which aryl is unsubstituted
or substituted,
2. heteroaryl having from 5 to 14 ring members,
in
which heteroaryl is unsubstituted or
substituted,
3. heterocycle having from 5 to 12 ring
members, in
which heterocycle is unsubstituted or
substituted,
4. -(C~-C6)-alkyl, in which alkyl is substituted
or
unsubstituted
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5. -C(O)-R",
6. -C(O)-O-R" or
7 -C(O)-N(R")2, or
R8 and R9 form, together with the nitrogen atom and carbon atom to which
5 they are in each case bonded, a heterocyclic ring of the formula Ila,
N
~D~ ( ~ (lla)
A B
X~Y
in which D and Z are defined as in formula II,
A is nitrogen atom or the radical -CH2-,
B is oxygen atom, sulfur atom, nitrogen atom or the radical
-CHz-,
X is oxygen atom, sulfur atom, nitrogen atom or the radical
-CHZ-,
Y is absent or is oxygen atom, sulfur atom, nitrogen atom or
the radical -CH2-, or
X and Y together form a phenyl, 1,2-diazine, 1,3-diazine, or 1,4-
diazine radical,
where the ring system which is formed by N, A, X, Y, B and carbon atom
does not contain more than one oxygen atom, X is not oxygen atom, sulfur
atom or nitrogen atom when A is nitrogen atom, does not contain more
than one sulfur atom, and contains 1, 2, 3 or 4 nitrogen atoms, and where
an oxygen atom and a sulfur atom are not present simultaneously,
where the ring system which is formed by N, A, X, Y, B and carbon atom is
unsubstituted or is substituted, once, twice or three times, independently of
each other, by (C~-Ca)-alkyl, in which alkyl is unsubstituted or substituted,
once or two times, by
1.1. -OH,
1.2. -(C~-Ca)-alkoxy,
1.3. halogen,
1.4. -N02,
1.5. -NH2,
1.6. -CF3,
1.7. methylenedioxyl,
1.8 -C(O),
1.9. -C(O)-CH3,
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6
1.10. -(C~-C4)-alkoxycarbonyl,
1.11. -CN,
1.12. -C(O)-OH,
1.13. -C(O)-NH2,
1.14. tetrazolyl,
1.15. phenyl,
1.16. phenoxy,
1.17. benzyl or
1.18. benzyloxy or
R9 and Z form, together with the carbon atoms to which they are in each
case bonded, a heterocyclic ring of the formula Ilc,
0
N R"
(Ilc)
Ra Tw/W
in which D, R8 and R~~ are defined as in formula II,
T is oxygen atom, sulfur atom, nitrogen atom or the radical
_CHz_~
W is oxygen atom, sulfur atom, nitrogen atom or the radical
-CH2-,
V is absent or is oxygen atom, sulfur atom, nitrogen atom or
the radical -CH2-, or
T and V or V and W together form a phenyl, 1,2-diazine, 1,3-
diazine or 1,4-diazine radical,
where the ring system which is formed by N, T, V, W and two carbon
atoms does not contain more than one oxygen atom, does not
contain more than one sulfur atom and contains 1, 2, 3 or 4 nitrogen
atoms, where an oxygen atom and a sulfur atom are not present
simultaneously, and where the ring system which is formed by N, T,
V, W and two carbon atoms is unsubstituted or is substituted, once,
twice or three times, independently of each other, by the
substituents which are defined above under 1.1. to 1.18., and
the other substituents R1, R2, R3 and R4 in each case are, independently of
each other,
1. hydrogen atom,
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w 7
2. halogen,
3. -(C~-C6)-alkyl,
4. heteroaryl having from 5 to 14 ring members, in
which heteroaryl
is unsubstituted or substituted,
5. heterocycle having from 5 to 12 ring members, in
which
heterocycle is unsubstituted or substituted,
6. -N 02,
7. -CN,
8. -O-(Co-C4)-alkylaryl,
9 -O-(C~-C4)-alkyl,
10 .-O-R",
11 .-N(R")2,
12 .-S(O)S-R", in which r is the integer zero, 1 or
2, or
13 . -CF3,
R5 is hydrogen atom,
1.
2. -OH or
3. =O, and
R6 is aryl, in which aryl is unsubstituted or substituted,
1.
2. phenyl which is substituted once or twice by
2.1 -CN,
2.2 -N02,
2.3 -O-(C~-G4)-alkyl,
2.4 -N(R")2,
2.5 -NH-C(O)-R",
2.6 -S(O)S R", in which s is the integer zero,
1 or 2,
2.7 -C(O)-R" or
2.8 -(C~-C4)-alkyl-NH2,
3. heteroaryl having from 5 to 14 ring members, is
unsubstituted or
is substituted once, twice or three times, or
4. heterocycle having from 5 to 12 ring members, is
unsubstituted
or is substituted once, twice or three times.
The invention furthermore relates to the use, according to the invention, of
the compound of the formula I, where
E is N atom or the radical -C(R'9)-,
where R'9 is hydrogen atom or the radical R9,
one of the substituents R', R2, R3 and R4 is a radical of the formula II, in
which
D is -C(O)-, -S(O)- or -S(O)2-,
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Ra is hydrogen atom or -(C~-C4)-alkyl,
R9 is 1. a characteristic radical of an amino acid which is derived from a
naturally occurring a-amino acid of the group alanine, valine,
leucine, isoleucine, phenylalanine, tyrosine, tryptophan, serine,
threonine, cysteine, methionine, asparagine, glutamine, lysine,
histidine, arginine, glutamic acid and aspartic acid,
2. a characteristic radical of an amino acid which is derived from an
amino acid which is not naturally occurring, such as 2-amino-
adipic acid, 2-aminobutyric acid, 2-aminoisobutyric acid,
2,4-diaminobutyric acid, 2,3-diaminopropionic acid,
1,2,3,4,-tetrahydroisoquinoline-1-carboxylic acid, 1,2,3,4-tetra-
hydroisoquinoline-3-carboxylic acid, 2-aminopimelic acid, 3-(2-
thienyl)alanine, 3-(3-thienyl)alanine, sarcosine, pipecolic acid,
2-aminoheptanoic acid, hydroxylysine, N-methylisoleucine, 6-N-
methyllysine, norleucine, N-methylvaline, norvaline, ornithine,
alto-isoleucine, 4-hydroxyproline, alto-hydroxylysine, allo-
threonine, 3-hydroxyproline, 3-(2-naphthyl)alanine, 3-(1-
naphthylalanine), homocysteine, homophenylalanine, homo-
cysteic acid, 2-amino-3-phenylaminoethylpropionic acid,
2-amino-3-phenylaminopropionic acid, homotryptophan, cysteic
acid, 3-(2-pyridyl)alanine, 3-(3-pyridyl)alanine, 3-(4-pyridyl)-
alanine, phosphinothricin, 4-fluorophenylalanine, 3-fluoro-
phenylalanine, 2-fluorophenylalanine, 4-chlorophenylalanine,
4-nitrophenylalanine, cyclohexylalanine, 4-aminophenylalanine,
citrulline, 5-fluorotryptophan, 5-methoxytryptophan, methionine
sulfone, methionine sulfoxide or -NH-NR"-CON(R~~)2, in which
R'1 is defined as below,
3. aryl, from the group anthryl, biphenylyl, 2-biphenylyl,
3-biphenylyl, 4-biphenylyl, fluorenyl, naphthyl, 1-naphthyl,
2-naphthyl or phenyl, in which aryl is unsubstituted or substituted
once, twice or three times by identical or different radicals from
the series -C(O)-(C~-C4)-alkyl, -C(O), =O, -NH-(C~-C4)-alkyl,
-NH-((C~-C4)-alkyl)2, -(C~-C$)-alkyl, -(C~-Ca)-alkoxy, halogen,
vitro, amino, trifluoromethyl, hydroxyl, -CF3, hydroxy-(C~-C4)-
alkyl, such as hydroxymethyl or 1-hydroxyethyl or 2-hydroxyethyl,
methylenedioxy, ethylenedioxy, formyl, acetyl, cyano,
hydroxycarbonyl, aminocarbonyl, -(C~-C4)-alkoxycarbonyl,
phenyl, phenoxy, benzyl, benzyloxy, -S(O)X R~~ in which x is the
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integer zero, 1 or 2, -O-(C~-C4)-alkyl, -C(O)-OH, -C(O)-O-(C~-C4)-
alkyl, -NH-C(O)-(C~-C4)-alkyl or tetrazolyl,
4. heteroaryl having from 5 to 14 ring members, in which heteroaryl
is unsubstituted or substituted and, as a radical, is derived from
the group azepine, azetidine, benzimidazole, benzodioxolane,
2-benzofuran, benzothiazole, benzothiophene, 2-benzo-
thiophene, 2-benzoxazole, [i-carboline, quinoxaline, quinazoline,
quinoline, 2-quinoxaline, cyclohepta[b]-5-pyrrole, diazepine,
dihydropyridine, 3-hydroxypyrro-2,4-dione, imidazole,
4-imidazole, imidazolidine, imidazoline, indazole, indole,
isoquinoline, isoindole, isothiazole, isothiazolidine, isoxazole,
2-isoxazolidine, isoxazolidine, isoxazolone, methylimidazole,
3-(N-methylpyrrolidine), morpholine, oxazole, 1,3,4-oxadiazole,
oxadiazolidinedione, oxadiazolone, 5-oxo-4,5-dihydro-
[1,3,4]oxadiazole, 5-oxo-1,2,4-thiadiazole, 1,2,3,5-oxathia-
diazole-2-oxide, 1-oxo-1,2-dihydro-3-isoquinol, phenylpyrrole,
5-phenyl-2-pyrrole, phthalazine, piperazine, piperidine, pyrazine,
pyrazole, pyrazoline, pyrazolidine, pyrazoline, pyridazine,
pyrimidine, pyridine, pyridyl-N-oxide, 2-pyrrole, 3-pyrrole,
pyrrolidine, pyrroline, 4,5,6,7-tetrahydro-2-indole, tetrahydro-
thienyl, tetrazole, thiadiazole, thiazole, thiomorpholine,
thiophene, triazole, triazolone or triazole,
in which heteroaryl is unsubstituted or substituted once, twice or
three times by identical or different radicals which are derived
from the series -C(O)-(C~-C4)-alkyl, -C(O), =O, -NH-(C~-C4)
alkyl, -NH-((C~-C4)-alkyl)2, -(C~-C$)-alkyl, -(C~-C8)-alkoxy,
halogen, nitro, amino, trifluoromethyl, hydroxyl, -CF3, hydroxy-
(C~-C4)-alkyl such as hydroxymethyl or 1-hydroxyethyl or
2-hydroxyethyl, methylenedioxy, ethylenedioxy, formyl, acetyl,
cyano, hydroxycarbonyl, aminocarbonyl, -(C~-C4)-alkoxycarbonyl,
phenyl, phenoxy, benzyl, benzyloxy, -S(O)X R", in which x is the
integer zero, 1 or 2, -O-(C~-C4)-alkyl, -C(O)-OH, -C(O)-O-(C~-C4)-
alkyl, -NH-C(O)-(C1-C4)-alkyl or tetrazolyl,
5. -(C~-C6)-alkyl in which alkyl is straight-chain or branched and is
unsubstituted or substituted once, twice or three times,
independently of each other, by
5.1 aryl, in which aryl is defined as above,
5.2 heteroaryl having from 5 to 14 ring members, in which
heteroaryl is defined as above,
CA 02495455 2005-02-15
5.3 -(C3-C6)-cycloalkyl,
5.4 -O-R" ,
5.5 =O,
5.6 halogen,
5 5.7 -CN,
5.8 -CF3,
5.9 -S(O)XR", in which x is the integer zero, 1 or 2,
5.10 -C(O)-O-R",
5.11 -C(O)-N(R")2,
10 5.12 -C(O)-R",
5.13 -N (R" )2,
5.14 a radical of the formula
R"
R"
R"
or
5.15 a radical of the formula R11
in which
R" is a) hydrogen atom,
b) (C~-C6)-alkyl in which alkyl is unsubstituted or
substituted once, twice or three times by
1. aryl, in which aryl is defined as above,
2. heteroaryl having 5 to 14 ring members, in
which heteroaryl is defined as above,
3. halogen,
4. -N-(C~-C6)~-alkyl, in which n is the integer zero,
1 or 2 and alkyl is unsubstituted or substituted
once, twice or three times, independently of
each other, by halogen or by -C(O)-OH,
5. -O-(C~-C6)-alkyl or
6. -C(O)-OH,
c) aryl, in which aryl is defined as above, or
d) heteroaryl having from 5 to 14 ring members, in
which heteroaryl is defined as above, and
in the case of (R")2, the radical R" has, independent of
each other, the meaning of a) to d),
Z is 1. aryl in which aryl is defined as above,
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2. heteroaryl having from 5 to 14 ring members, in which heteroaryl
is defined as above,
3. -(C~-C6)-alkyl, in which alkyl is straight-chain or branched and is
substituted once or twice by phenyl or -OH,
4. -C(O)-O-R~~, or
5. -C(O)-N(R")2, and
the other substituents R', R2, R3 and R4 in each case are, independently of
each other,
1. hydrogen atom,
2. halogen,
3. -(C~-C4)-alkyl,
4. heteroaryl having from 5 to 14 ring members, in which heteroaryl
is as defined above,
5. -(C~-C6)-alkyl,
6. -N02,
7. -CN,
8. -O-(Co-C4)-alkyl-aryl, in which aryl is defined as above,
9. -O-(C~-C4)-alkyl,
10.-OR's,
11. -N(R~ ~ )2,
12.-S(O)X R~~, in which x is the integer zero, 1 or 2, or
13. -CF3,
R5 is 1. hydrogen atom,
2. -OH, or
3. =O, and
R6 is 1. aryl, from the group naphthyl, 1-naphthyl, 2-naphthyl, phenyl,
biphenylyl, 2-biphenylyl, 3-biphenylyl, 4-biphenylyl, anthryl or
fluorenyl,
in which aryl is unsubstituted or substituted, once, twice or three
times, by identical or different radicals from the series -C(O)-(C~-Ca)
alkyl, -C(O), =O, -NH-(C~-C4)-alkyl, -NH-((C~-C4)-alkyl)2, -(C~-Cg)
alkyl, -(C~-C$)-alkoxy, halogen, nitro, amino, trifluoromethyl,
hydroxyl, -CF3, hydroxy-(C~-C4)-alkyl such as hydroxymethyl or
1-hydroxyethyl or 2-hydroxyethyl, methylenedioxy, ethylenedioxy,
formyl, acetyl, cyano, hydroxycarbonyl, aminocarbonyl, -(C~-C4)-
alkoxycarbonyl, phenyl, phenoxy, benzyl, benzyloxy, -S(O)X R~~, in
which x is the integer zero, 1 or 2, -O-(C~-C4)-alkyl, -C(O)-OH,
-C(O)-O-(C~-C4)-alkyl, -NH-C(O)-(C~-C4)-alkyl or tetrazolyl, or
CA 02495455 2005-02-15
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2. heteroaryl having from 5 to 14 ring members, in which heteroaryl
is defined as above and
in which heteroaryl is unsubstituted or substituted, once, twice or
three times, by identical or different radicals from the series
-C(O)-(C~-C4)-alkyl, -C(O), =O, -NH-(C~-C4)-alkyl, -NH-((C~-C4)-
alkyl)2, -(C~-C$)-alkyl, -(C~-C8)-alkoxy, halogen, nitro, amino,
trifluoromethyl, hydroxyl, -CF3, hydroxy-(C~-C4)-alkyl such as
hydroxymethyl or 1-hydroxyethyl or 2-hydroxyethyl,
methylenedioxy, ethylenedioxy, formyl, acetyl, cyano,
hydroxycarbonyl, aminocarbonyl, -(C~-C4)-alkoxycarbonyl,
phenyl, phenoxy, benzyl, benzyloxy, -S(O)X R", in which x is the
integer zero, 1 or 2, -O-(C,-C4)-alkyl, -C(O)-OH,
-C(O)-O-(C~-C4)-alkyl, -NH-C(O)-(C~-C4)-alkyl or tetrazolyl.
The invention furthermore relates to the use, according to the invention, of
the compound of the formula I, where
E is N atom or the radical -C(R'9)-,
in which R'9 is hydrogen atom,
one of the substituents R', R2, R3 and R4 is a radical of the formula II, in
which
R8 is hydrogen atom,
R9 is 1. a characteristic radical of an amino acid from the group histidine,
serine, tryptophan, threonine, cysteine, methionine, asparagine,
glutamine, lysine, arginine, glutamic acid and aspartic acid, or
2. -(C~-C6)-alkyl, in which alkyl is straight-chain or branched and is
unsubstituted or substituted, once or twice, by
a) phenyl,
b) a radical from the group azepine, azetidine, benzimidazole,
benzothiazole, benzothiophene, benzoxazole, diazepine,
imidazole, indole, isothiazole, isoxazole, morpholine, 1,3,4-oxa
diazole, 5-oxo-4,5-dihydro-[1,3,4]oxadiazole, oxazole piperidine,
pyrazine, pyrazole, pyridine, pyrimidine, pyrrole, pyrrolidine,
pyrroline, thiazole, thiomorpholine, thiophene or triazole,
c) -NH(R"),
d) -C(O)-R'2, in which
R'2 is naphthyl, phenyl, morpholinyl or pyrimidinyl,
e) -O-R",
f) -N(R'2)-phenyl, in which R'2 is defined as above,
g) -S(O)X R'2, in which x is zero, 1 or 2, and
CA 02495455 2005-02-15
13
h) -CN, or
i) -(C3-C6)-cycloalkyl,
and the radicals defined above by a), b), d) and i) and R'2 are
unsubstituted or are substituted, once or twice, by -OH, -(C~-C4)
alkyl, -CF3, halogen, -O-(C~-C4)-alkyl, -COOH, -C(O)-O-(C~-C4)
alkyl, -NH2 or -NH-C(O)-(C~-C4)-alkyl,
Z is 1. a heteroaryl radical from the group 3-hydroxypyrro-2,4-dione,
imidazole, imidazolidine, imidazoline, indazole, isothiazole,
isothiazolidine, isoxazole, isoxazolidine, 2-isoxazolidine,
isoxazolone, morpholine, 1,3,4-oxadiazole, oxadiazolidinedione,
oxadiazolone, 1,2,3,5-oxathiadiazole-2-oxide, oxazole, 5-oxo-4,5-
dihydro-[1,3,4]oxadiazole, 5-oxo-1,2,4-thiadiazole, piperazine,
pyrazine, pyrazole, pyrazolidine, pyrazoline, pyridazine, pyrimidine,
tetrazole, thiadiazole, thiazole, thiomorpholine, triazole or triazolone,
and
the heteroaryl radical is unsubstituted or substituted, once, twice
or three times, independently of each other, by
1.1 -C(O)-R'S, in which R'S is hydrogen atom or -(C~-C4)-alkyl,
1.2 -(C~-C4)-alkyl,
1.3 -O-R'S, in which R'5 is hydrogen atom or -(C~-C4)-alkyl,
1.4 -N(R'S)-R'6, in which R'S and R'6 are, independently of
each other, hydrogen atom or -(C~-C4)-alkyl,
1.5 halogen, or
1.6 keto radical,
2. -C(O)-R'5, in which R'S is hydrogen atom or -(C~-C4)-alkyl,
3. -C(O)-R'S, in which R'S is hydrogen atom or -(C~-C4)-alkyl, or
4. -C(O)-N(R'S)-R's, in which R'5 and R'6 are, independently of
each other, hydrogen atom or -(C~-C4)-alkyl,
R" is 1. -(C~-C4)-alkyl,
2. R'3 or,
3. -N(R'3)2,
in which R'3 is, independently of each other,
a) hydrogen atom,
b) -(C~-C6)-alkyl,
c) -(C~-C4)-alkyl-O-(C~-C4)-alkyl,
d) -(C~-C6)-alkyl-N(R'S)2, in which R'5 is defined as above, or
e) -(Co-C4)-alkyl which is substituted, once or twice, by
imidazolyl, morpholinyl or phenyl, or
CA 02495455 2005-02-15
14
Rs and R9 form, together with the nitrogen atom and carbon atom to which
they are in each case bonded, a ring of the formula Ila from the
group pyrrole, pyrroline, pyrrolidine, pyridine, piperidine, piperylene,
pyridazine, pyrimidine, pyrazine, piperazine, pyrazole, imidazole,
pyrazoline, imidazoline, pyrazolidine, imidazolidine, oxazole,
isoxazole; 2-isoxazolidine, isoxazolidine, morpholine, isothiazole,
thiazole, tetrazole, 1,2,3,5-oxathiadiazole-2-oxides, oxadiazolone,
isoxazolone, triazolones, oxadiazolidinedione, triazole, which are
substituted by F, CN, CF3 or COO-(C~-C4)-alkyl, 3-hydroxypyrro-2,4-
diones, 5-oxo-1,2,4-thiadiazoles, 1,3,4-oxadiazole, isothiazolidine,
thiomorpholine, indazole, thiadiazole, benzimidazole, quinoline,
triazole, phthalazine, quinazoline, quinoxaline, purine, pteridine,
indole, tetrahydroquinoline, tetrahydroisoquinoline and isoquinoline,
or
R9 and Z form, together with the carbon atoms to which they are in each
case bonded, a ring of the formula Ilc from the group pyrrole,
pyrroline, pyrrolidine, pyridine, piperidine, piperylene, pyridazine,
pyrimidine, pyrazine, piperazine, pyrazole, imidazole, pyrazoline,
imidazoline, pyrazolidine, imidazolidine, oxazole, isoxazole,
2-isoxazolidine, isoxazolidine, morpholine, isothiazole, thiazole,
isothiazolidine, thiomorpholine, indazole, thiadiazole, benzimidazole,
quinoline, triazole, phthalazine, quinazoline, quinoxaline, purine,
pteridine, indole, tetrahydroquinoline, tetrahydroisoquinoline,
isoquinoline, tetrazole, 1,2,3,5-oxathiadiazole-2-oxides,
oxadiazolone, isoxazolone, triazolones, oxadiazolidinedione,
triazole, which are substituted by F, CN, CF3 or COO-(C~-C4)-alkyl,
3-hydroxypyrro-2,4-diones, 1,3,4-oxadiazole and 5-oxo-1,2,4-
thiadiazole and
the other substituents R', R2, R3 and R4 in each case are, independently of
each other,
1. hydrogen atom,
2. halogen,
3. -(C~-C4)-alkyl,
4. -CN,
5. -N02,
6. -O-(Co-C4)-alkyl-phenyl,
7. -O-(C~-C4)-alkyl,
8. -N-(Co-C4)-alkyl-phenyl,
9. -N-(C~-C4)-alkyl or
CA 02495455 2005-02-15
10. -CF3,
R5 is 1. hydrogen atom,
2. -OH, or
3. =O, and
5 R6 is 1. phenyl, substituted, once or twice, by
1.1 -CN,
1.2 -N02,
1.3 -O-(C~-C4)-alkyl, or
1.4 -NH2, or
10 2. is pyridine or pyrimidine,
where pyridine or pyrimidine is unsubstituted or substituted, once,
twice or three times, by identical or different radicals from the series
-C(O)-(C~-C4)-alkyl, -C(O), =O, -NH-(C~-C4)-alkyl, -NH-((C~-C4)-
alkyl)2, -(C~-C$)-alkyl, -(C~-C$)-alkoxy, halogen, nitro, amino,
15 trifluoromethyl, hydroxyl, -CF3, hydroxy-(C~-C4)-alkyl such as
hydroxymethyl or 1-hydroxyethyl or 2-hydroxyethyl, methylenedioxy,
ethylenedioxy, formyl, acetyl, cyano, hydroxycarbonyl,
aminocarbonyl, -(C~-C4)-alkoxycarbonyl, phenyl, phenoxy, benzyl,
benzyloxy, -S(O)x R", in which x is the integer zero, 1 or 2,
-O-(C~-C4)-alkyl, -C(O)-OH, -C(O)-O-(C~-C4)-alkyl,
-NH-C(O)-(C~-C4)-alkyl or tetrazolyl.
The invention furthermore relates to the use, according to the invention, of
the compound of the formula I,
where E is the radical -C(R'9)-,
in which R'9 is hydrogen atom or R9,
one of the substituents R', R2, R3 and R4 is a radical of the formula II in
which
D is -C(O)-,
Ra is hydrogen atom,
Z is 5-oxo-4,5-dihydro-[1,3,4]oxadiazole, -C(O)-OH or -C(O)-NH2,
R9 is 1. -(C~-C4)-alkyl, in which alkyl is straight-chain or branched and is
substituted once or twice, independently of each other, by
1.1 -S(O)-R", where R" is defined as below,
1.2 -N(R")2, where R" is defined as below, or
1.3 pyrrole, or
2. the characteristic radical of an amino acid from the group
histidine, tryptophan, serine, threonine, cysteine, methionine,
CA 02495455 2005-02-15
16
asparagine, glutamine, lysine, arginine, glutamic acid and
aspartic acid,
R" is a) hydrogen atom,
b) -(C~-C6)-alkyl, in which alkyl is unsubstituted or substituted, once
or three times, independently of each other, by halogen, or
c) phenyl, in which phenyl is unsubstituted or substituted, once to
three times, independently of each other, by halogen or -(C~-C4)-
alkyl,
the other substituents R', R2, R3 and R4 are in each case hydrogen atom,
R5 is hydrogen atom, and
R6 is phenyl, pyridine or pyrimidine,
where phenyl, pyridine or pyrimidine is unsubstituted or substituted,
once, twice or three times, by identical or different radicals from the
series -C(O)-(C~-C4)-alkyl, -C(O), =O, -NH-(C~-C4)-alkyl,
-NH-((C~-C4)-alkyl)z, -(C~-C8)-alkyl, -(C1-C8)-alkoxy, halogen, nitro,
amino, trifluoromethyl, hydroxyl, -CF3, hydroxy-(C~-C4)-alkyl such as
hydroxymethyl or 1-hydroxyethyl or 2-hydroxyethyl, methylenedioxy,
ethylenedioxy, formyl, acetyl, cyano, hydroxycarbonyl,
aminocarbonyl, -(C~-C4)-alkoxycarbonyl, phenyl, phenoxy, benzyl,
benzyloxy, -S(O)X R", in which x is the integer zero, 1 or 2,
-O-(C~-C4)-alkyl, -C(O)-OH, -C(O)-O-(C~-C4)-alkyl,
-NH-C(O)-(C~-C4)-alkyl or tetrazolyl.
The invention furthermore relates to the use, according to the invention, of
the compound of the formula la
R21
R24
JN
O
R31
R22\N I \ \ N (la)
/ N M-
H /N-H
R23
andlor a stereoisomeric form of the compound of the formula la and/or a
physiologically tolerated salt of the compound of the formula la, where
CA 02495455 2005-02-15
17
E and M are identical or different and are, independently of each other N
atom or CH
R21 and R31 are identical or different and are, independently of each
other,
1. hydrogen atom,
2. halogen,
3. -(C~-C4)-alkyl,
4. -CN,
5. -CF3,
6. -OR'S, in which R'S is hydrogen atom or -(C~-C4)-alkyl,
7. -N(R'S)-R'6, in which R'S and R'6 are, independently
of each
other, hydrogen atom or -(C~-C4)-alkyl,
8. -C(O)-R'5, in which R'S is hydrogen atom or -(C~-C4)-alkyl,
or
9. -S(O)X R'S, in which x is the integer zero, 1
or 2, and R'~ is
hydrogen atom or -(C~-C4)-alkyl,
R22 is 1. a heteroaryl radical from the group 3-hydroxypyrro-2,4-dione,
imidazole, imidazolidine, imidazoline, indazole,
isothiazole,
isothiazolidine, isoxazole, 2-isoxazolidine, isoxazolidine,
isoxazolone, morpholine, oxazole, 1,3,4-oxadiazole,
oxadiazolidinedione, oxadiazolone, 1,2,3,5-oxathiadiazole-2-
oxide, 5-oxo-4,5-dihydro-[1,3,4]oxadiazole, 5-oxo-1,2,4-
thiadiazole, piperazine, pyrazine, pyrazole, pyrazoline,
pyrazolidine, pyridazine, pyrimidine, tetrazole,
thiadiazole,
thiazole, thiomorpholine, triazole or triazolone,
and
the heteroaryl radical is unsubstituted or is
substituted once,
twice or three times, independently of each other,
by
1.1 -C(O)-R'5, in which R'S is hydrogen atom or
-(C~-C4)-
alkyl,
1.2 -(C~-C4)-alkyl,
1.3 -O-R'S, in which R'5 is hydrogen atom or -(C~-C4)-alkyl,
1.7 -N(R'S)-R'6, in which R'S and R'6 are, independently
of
each other, hydrogen atom or -(C~-C4)-alkyl,
1.8 halogen, or
1.9 keto radical,
2. -C(O)-R'5, in which R'S is hydrogen atom or -(C~-C4)-alkyl,
3. -C(O)-OR'5, in which R'S is hydrogen atom or -(C,-C4)-alkyl,
or
4. -C(O)-N(R'~)-R'$, in which R" and R'$ are, independently
of
each other, hydrogen atom, -(C~-C4)-alkyl-OH,
-O-(C~-C4)-alkyl or
-(C~-C4)-alkyl,
CA 02495455 2005-02-15
18
R23 is hydrogen atom or-(C~-C4)-alkyl,
R24 is 1. a heteroaryl radical from the group pyrrole, furan, thiophene,
imidazole, pyrazole, oxazole, isoxazole, thiazole, isothiazole,
tetrazole, 1,2,3,5-oxathiadiazole-2-oxides, triazolones, oxa-
diazolones, isoxazolones, oxadiazolidinediones, triazoles,
3-hydroxypyrro-2,4-diones, 5-oxo-1,2,4-thiadiazoles, pyridine,
pyrazine, pyrimidine, indole, isoindole, indazole, phthalazine,
quinoline, isoquinoline, quinoxaline, quinazoline, cinnoline,
(3-carboline and benzo fused cyclopenta derivatives or cyclohexa
derivatives of these heteroaryl radicals,
where the heteroaryl radical is unsubstituted or is substituted,
once, twice or three times, independently of each other, by
-(C~-C5)-alkyl, -(C~-C5)-alkoxy, halogen, nitro, amino,
trifluoromethyl, hydroxyl, hydroxy-(C~-C4)-alkyl, methylenedioxy,
ethylenedioxy, formyl, acetyl, cyano, hydroxycarbonyl, amino-
carbonyl or -(C~-C4)-alkoxycarbonyl, or
2. an aryl radical from the group phenyl, naphthyl, 1-naphthyl,
2-naphthyl, biphenylyl, 2-biphenylyl, 3-biphenylyl and
4-biphenylyl, anthryl or fluorenyl, and
the aryl radical is unsubstituted or substituted, once, twice or
three times, independently of each other, by -(C~-C5)-alkyl,
-(C~-C5)-alkoxy, halogen, nitro, amino, trifluoromethyl, hydroxyl,
hydroxy-(C~-C4)-alkyl, methylenedioxy, ethylenedioxy, formyl,
acetyl, cyano, hydroxycarbonyl, aminocarbonyl or -(C~.C4)
alkoxycarbonyl.
The invention furthermore relates to the use, according to the invention, of
the compound of the formula la, where
E and M are identical or different and are, independently of each other, N
atom or CH,
R21 and R31 are identical or different and, independently of each other,
are defined as above under 1. to 9.,
R22 is 1. a heteroaryl radical from the group imidazole, isothiazole,
isoxazole, 2-isoxazolidine, isoxazolone, 1,3,4-oxadiazole,
oxadiazolidinedione, 1,2,3,5-oxadiazolone, oxazole, 5-oxo-4,5
dihydro-[1,3,4]oxadiazole, tetrazole, thiadiazole, thiazole, triazole
or triazolone, and the heteroaryl radical is unsubstituted or is
substituted once, twice or three times, independently of each
other, by
CA 02495455 2005-02-15
19
1.1 keto radical,
1.2 halogen or
1.3 -(C~-C2)-alkyl, or
2. -C(O)-N(R'~)-RIB, in which R~~ and R's are, independently of
each other, hydrogen atom, -(C~-C4)-alkyl-OH, -O-(C~-C4)-alkyl,
or -(C~-C4)-alkyl,
R23 is hydrogen atom, methyl or ethyl,
R24 is 1. a heteroaryl radical from the group of the unsaturated, partially
saturated or completely saturated rings which are derived from
pyridine, pyrazine, pyrimidine, pyridazine, pyrrole, furan,
thiophene, imidazole, pyrazole, oxazole, isoxazole, thiazole,
triazole or isothiazole,
where the heteroaryl radical is unsubstituted or substituted, once,
twice or three times, independently of each other, by -(C1-C4)
alkyl, -(C1-C4)-alkoxy, F, CI, J, Br, vitro, amino, trifluoromethyl,
hydroxyl, hydroxy-(C1-C4)-alkyl, methylenedioxy, ethylenedioxy,
formyl, acetyl, cyano, hydroxycarbonyl, aminocarbonyl or
-(C1-C4)-alkoxycarbonyl, or
2. phenyl and phenyl is unsubstituted or is substituted once, twice
or three times, independently of each other, by F, CI, I, Br, CF3,
-OH, -(C~-C4)-alkyl or -(C~-C4)-alkoxy.
The invention furthermore relates to the use, according to the invention, of
the compound N-[(S)-2-diphenylamino-1-(5-oxo-4,5-dihydro-
[1,3,4]oxadiazol-2-yl)ethyl]-2-(2-methylaminopyrimidin-4-yl)-1H-indole-5-
carboxamide or N-((S)-1-carbamoyl-2-diphenylaminoethyl)-2-(2-methyl-
aminopyrimidin-4-yl)-1 H-benzimidazole-5-carboxamide.
The term "halogen" is understood as meaning fluorine, chlorine, bromine or
iodine. The terms "-(C~-C8)-alkyl", "-(C~-C6)-alkyl" and "-(C~-C4)-alkyl" are
understood as meaning hydrocarbon radicals whose carbon chain is
straight-chain or branched and contains from 1 to 8, from 1 to 6 and from 1
to 4 carbon atoms, respectively, such as methyl, ethyl, propyl, butyl, tert-
butyl, pentyl, hexyl, heptyl or octyl. The term "-(Co)-alkyl" is understood as
meaning a covalent bond. Examples of cyclic alkyl radicals are 3- to
6-membered monocycles such as cyclopropyl, cyclobutyl, cyclopentyl and
cyclohexyl.
CA 02495455 2005-02-15
The phrase "R8 and R9 form, together with the nitrogen atom and carbon
atom to which they are in each case bonded, a heterocyclic ring of the
formula Ila" is understood as meaning radicals which are derived from
pyrrole, pyrroline, pyrrolidine, imidazole, pyrazole, oxazole, isoxazole,
5 tetrazole, isoxazoline, isoxazolidine, morpholine, thiazole, isothiazole,
isothiazoline, purine, isothiazolidine, thiomorpholine, pyridine, piperidine,
pyrazine, piperazine, pyrimidine, pyridazine, indole, isoindole, indazole,
benzimidazole, phthalazine, quinoline, isoquinoline, quinoxaline,
quinazoline, cinnoline, pteridine, triazolones, tetrazole, 1,2,3,5-oxathia-
10 diazole-2-oxides, oxadiazolones, isoxazolones, oxadiazolidinediones,
triazoles, which are substituted by F, -CN, -CF3 or -C(O)-O-(C~-C4)-alkyl, 3-
hydroxypyrro-2,4-diones, 5-oxo-1,2,4-thiadiazoles, imidazolidine, carboline
and benzofused derivatives of these heterocycles.
15 The phrase "R9 and Z form, together with the carbon atoms to which they
are in each case bonded, a heterocyclic ring of the formula Ilc" is
understood as meaning radicals which [lacuna] from the group pyrrole,
pyrroline, pyrrolidine, pyridine, piperidine, piperylene, pyridazine,
pyrimidine, pyrazine, piperazine, pyrazole, imidazole, pyrazoline,
20 imidazoline, pyrazolidine, imidazolidine, oxazole, isoxazole, 2-
isoxazolidine,
isoxazolidine, morpholine, isothiazole, thiazole, isothiazolidine,
thiomorpholine, indazole, thiadiazole, benzimidazole, quinoline, triazole,
phthalazine, quinazoline, quinoxaline, purine, pteridine, indole,
tetrahydroquinoline, tetrahydroisoquinoline, isoquinoline, tetrazole, 1,2,3,5-
oxathiadiazole-2-oxides, oxadiazolone, isoxazolone, triazolone,
3-hydroxypyrro-2,4-diones, 1,3,4-oxadiazole and 5-oxo-1,2,4-thiadiazole,
oxadiazolidinedione, triazole, which are unsubstituted or substituted by F,
CN, CF3 or C(O)-O-(C~-C4)-alkyl.
The phrase "heteroaryl radical from the group of the unsaturated, partially
saturated or completely saturated rings which are derived from pyridine,
pyrazine, pyrimidine, pyridazine, pyrrole, fu ran, thiophene, imidazole,
pyrazole, oxazole, isoxazole, thiazole and isothiazole", is understood as
meaning, for example, compounds such as piperazine, pyrazoline,
imidazoline, pyrazolidine, imidazolidine, tetrahydropyridine, isoxazoline,
isoxazolidine, morpholine, isothiazoline, isothiazolidine, tetrahydro-1,4-
thiazine and piperidine.
The term "aryl" is understood as meaning aromatic hydrocarbon radicals
having from 6 to 14 carbon atoms in the ring. Examples of -(C6-C14)-aryl
CA 02495455 2005-02-15
21
radicals are phenyl, naphthyl, for example 1-naphthyl and 2-naphthyl,
biphenylyl, for example 2-biphenylyl, 3-biphenylyl and 4-biphenylyl, anthryl
and fluorenyl. Biphenylyl radicals, naphthyl radicals and, in particular,
phenyl radicals are preferred aryl radicals. Aryl radicals, in particular
phenyl
radicals, can be substituted once or more than once, preferably once, twice
or three times, by identical or different radicals, preferably by radicals
from
the series -(C~-C$)-alkyl, in particular -(C~-C4)-alkyl, -(C~-C8)-alkoxy, in
particular -(C~-C4)-alkoxy, halogen, nitro, amino, trifluoromethyl, hydroxyl,
hydroxy-(C~-C4)-alkyl, such as hydroxymethyl or 1-hydroxyethyl or
2-hydroxyethyl, methylenedioxy, ethylenedioxy, formyl, acetyl, cyano,
hydroxycarbonyl, aminocarbonyl, -(C~-C4)-alkoxycarbonyl, phenyl,
phenoxy, benzyl, benzyloxy and tetrazolyl. The same applies, in a
corresponding manner, for example, for radicals such as arylalkyl or
arylcarbonyl. Arylalkyl radicals are, in particular, benzyl and also 1- and
2-naphthylmethyl, 2-, 3- and 4-biphenylylmethyl and 9-fluorenylmethyl.
Substituted arylalkyl radicals are, for example, benzyl radicals and
naphthylmethyl radicals which are substituted, in the aryl moiety, by one or
more -(C~-C$)-alkyl radicals, in particular -(C~-C4)-alkyl radicals, for
example 2-, 3- and 4-methylbenzyl, 4-isobutylbenzyl, 4-tert-butylbenzyl,
4-octylbenzyl, 3,5-dimethylbenzyl, pentamethylbenzyl, 2-, 3-, 4-, 5-, 6-, l-
and 8-methyl-1-naphthylmethyl, and 1-, 3-, 4-, 5-, 6-, 7- and 8-methyl-2-
naphthylmethyl, benzyl radicals and naphthylmethyl radicals which are
substituted, in the aryl moiety, by one or more -(C~-C$)-alkoxy radicals, in
particular -(C~-C4)-alkoxy radicals, for example 4-methoxybenzyl,
4-neopentyloxybenzyl, 3,5-dimethoxybenzyl, 3,4-methylenedioxybenzyl
and 2,3,4-trimethoxybenzyl, nitrobenzyl radicals, for example 2-, 3- and 4
nitrobenzyl, halobenzyl radicals, for example 2-, 3- and 4-chlorobenzyl, 2-,
3- and 4-fluorobenzyl, 3,4-dichlorobenzyl, pentafluorobenzyl and
trifluoromethylbenzyl radicals, for example 3- and 4-trifluoromethylbenzyl
and 3,5-bis(trifluoromethyl)benzyl.
In monosubstituted phenyl radicals, the substituent can be located in the 2
position, the 3 position or the 4 position. Doubly substituted phenyl can be
substituted in the 2,3 position, the 2,4 position, the 2,5 position, the 2,6
position, the 3,4 position or the 3,5 position. In triply substituted phenyl
radicals, the substituents can be located in the 2,3,4 position, the 2,3,5
position, the 2,4,5 position, the 2,4,6 position, the 2,3,6 position or the
3,4,5
position.
CA 02495455 2005-02-15
22
The comments made with regard to the aryl radicals apply, in a
corresponding manner, to divalent arylene radicals, for example to
phenylene radicals, which can be present, for example, as 1,4-phenylene
or as 1,3-phenylene. Phenylene-(C~-C6)-alkyl is, in particular,
phenylenemethyl (-C6H4-CH2-) and phenyleneethyl, (C~-C6)-
alkylenephenyl, in particular methylenephenyl (-CH2-C6H4-). Phenylene-
(C2-C6)-alkenyl is, in particular, phenyleneethenyl and phenylenepropenyl.
The phrase "heteroaryl having from 5 to 14 ring members" means a radical
of a monocyclic or polycyclic aromatic system having from 5 to 14 ring
members which contains 1, 2, 3, 4 or 5 heteroatoms as ring members.
Examples of heteroatoms are N, O and S. If several heteroatoms are
present, they may be identical or different. Heteroaryl radicals can also be
substituted, once or more than once, preferably once, twice or three times,
by identical or different radicals from the series ---(C~-C8)-alkyl, in
particular
-(C~-C4)-alkyl, -(C~-C8)-alkoxy, in particular -(C~-C4)-alkoxy, halogen,
nitro,
-N(R'°)2, trifluoromethyl, hydroxyl, hydroxy-(C~-C4)-alkyl, such as
hydroxymethyl or 1-hydroxyethyl or 2-hydroxyethyl, methylenedioxy, formyl,
acetyl, cyano, hydroxycarbonyl, aminocarbonyl, -(C~-C4)-alkoxycarbonyl,
phenyl, phenoxy, benzyl, benzyloxy and tetrazolyl. Heteroaryl having from
5 to 14 ring members is preferably a monocyclic or bicyclic aromatic radical
which contains 1, 2, 3 or 4, in particular 1, 2 or 3, identical or different
heteroatoms from the series N, O and S, and which can be substituted by
1, 2, 3 or 4, in particular 1 to 3, identical or different substituents from
the
series -(C~-C6)-alkyl, -(C~-C6)-alkoxy, fluorine, chlorine, nitro, -N(R~o)2,
trifluoromethyl, hydroxyl, hydroxy-(C~-C4)-alkyl, -(C~-C4)-alkoxycarbonyl,
phenyl, phenoxy, benzyloxy and benzyl. Particularly preferably, heteroaryl
is a monocyclic or bicyclic aromatic radical having from 5 to 10 ring
members, in particular a 5-membered to 6-membered monocyclic aromatic
radical which contains 1, 2 or 3, in particular 1 or 2, identical or different
heteroatoms from the series N, O and S and which can be substituted by 1
or 2 identical or different substituents from the series -(C~-C4)-alkyl,
halogen, hydroxyl, -N(R'°)2, -(C~-C4)-alkoxy, phenyl, phenoxy,
benzyloxy
and benzyl.
The term "heterocycle having from 5 to 12 ring members" means a
monocyclic or bicyclic 5-membered to 12-membered heterocyclic ring
which is partially saturated or completely saturated. Examples of
heteroatoms are N, O and S. The heterocycle is unsubstituted or is
CA 02495455 2005-02-15
23
substituted by identical or different substituents at one or more carbon
atoms or at one or more heteroatoms. These substituents have been
defined above in connection with the heteroaryl radical. In particular, the
heterocyclic ring is substituted at carbon atoms, once or more than once,
for example once, twice, three times or four times, by identical or different
radicals from the series -(C~-C8)-alkyl, for example -(C~-C4)-alkyl, -(C~-C8)-
alkoxy, for example -(C~-C4)-alkoxy, such as methoxy, phenyl-(C~-C4)-
alkoxy, for example benzyloxy, hydroxyl, oxo, halogen, nitro, amino or
trifluoromethyl and/or is substituted at the ring nitrogen atoms) in the
heterocyclic ring by -(C~-C8)-alkyl, for example -(C~-C4)-alkyl such as
methyl or ethyl, by optionally substituted phenyl or phenyl-(C~-C4)-alkyl, for
example benzyl. Nitrogen heterocycles can also be present as N-oxides or
as quaternary salts.
Examples of the terms heteroaryl having from 5 to 14 ring members and
heterocycle having from 5 to 12 ring members are radicals which are
derived from pyrrole, furan, thiophene, imidazole, pyrazole, oxazole,
isoxazole, thiazole, isothiazole, tetrazole, 1,2,3,5-oxathiadiazole-2-oxides,
triazolones, oxadiazolones, isoxazolone, oxadiazolidinedione, triazole,
which are substituted by F, -CN, -CF3 or -C(O)-O-(C~-C4)-alkyl,
3-hydroxypyrro-2,4-diones, 5-oxo-1,2,4-thiadiazoles, pyridine, pyrazine,
pyrimidine, indole, isoindole, indazole, phthalazine, quinoline, isoquinoline,
quinoxaline, quinazoline, cinnoline, -carboline and benzo fused, cyclopenta
fused, cyclohexa fused or cyclohepta fused derivatives of these
heterocycles. Particular preference is given to the radicals 2- or 3-pyrrolyl,
phenylpyrrolyl, such as 4- or 5-phenyl-2-pyrrolyl, 2-furyl, 2-thienyl,
4-imidazolyl, methylimidazolyl, for example 1-methyl-2-, -4- or -5-imidazolyl,
1,3-thiazol-2-yl, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-, 3- or 4-pyridyl-N-
oxide,
2-pyrazinyl, 2-, 4- or 5-pyrimidinyl, 2-, 3- or 5-indolyl, substituted 2-
indolyl,
for example 1-methyl-, 5-methyl-, 5-methoxy-, 5-benzyloxy-, 5-chloro- or
4,5-dimethyl-2-indolyl, 1-benzyl-2- or -3-indolyl, 4,5,6,7-tetrahydro-2-
indolyl,
cyclohepta[b]-5-pyrrolyl, 2-, 3- or 4-quinolyl, 1-, 3- or 4-isoquinolyl,
1-oxo-1,2-dihydro-3-isoquinolyl, 2-quinoxalinyl, 2-benzofuranyl, 2-benzo-
thienyl, 2-benzoxazolyl or benzothiazolyl or dihydropyridinyl, pyrrolidinyl,
for
example 2- or 3-(N-methylpyrrolidinyl), piperazinyl, morpholinyl,
thiomorpholinyl, tetrahydrothienyl and benzodioxolanyl.
The general structural formula of a,-amino acids is as follows:
CA 02495455 2005-02-15
- 24
RFC-COOH
H.,N
The a-amino acids differ from each other in the radical R which, within the
context of the present application, is designated the "characteristic radical"
of an amino acid. When R9 denotes the characteristic radical of amino acid,
use is preferably made of the characteristic radicals of the following
naturally occurring a-amino acids: glycine, alanine, valine, leucine,
isoleucine, phenylalanine, tyrosine, tryptophan, serine, threonine, cysteine,
methionine, asparagine, glutamine, lysine, histidine, arginine, glutamic acid
and aspartic acid. Particular preference is given to histidine, tryptophan,
serine, threonine, cysteine, methionine, asparagine, glutamine, lysine,
arginine, glutamic acid and aspartic acid. In addition, amino acids which do
not occur naturally, such as 2-aminoadipic acid, 2-aminobutyric acid,
2-aminoisobutyric acid, 2,3-diaminopropionic acid, 2,4-diaminobutyric acid,
1,2,3,4-tetrahydroisoquinoline-1-carboxylic acid, 1,2,3,4-tetrahydroiso-
quinoline-3-carboxylic acid, 2-aminopimellic acid, phenylglycine, 3-(2-
thienyl)alanine, 3-(3-thienyl)alanine, 2-(2-thienyl)glycine, 2-aminoheptanoic
acid, pipecolic acid, hydroxylysine, sarcosine, N-methylisoleucine, 6-N-
methyllysine, N-methylvaline, norvaline, norleucine, ornithine,
alloisoleucine, allothreonine, allohydroxylysine, 4-hydroxyproline,
3-hydroxyproline, 3-(2-naphthyl)alanine, 3-(1-naphthylalanine),
homophenylalanine, homocysteine, homocysteic acid, homotryptophan,
cysteic acid, 3-(2-pyridyl)alanine, 3-(3-pyridyl)alanine, 3-(4-
pyridyl)alanine,
2-amino-3-phenylaminopropionic acid, 2-amino-3-phenylaminoethyl-
propionic acid, phosphinothricin, 4-fluorophenylalanine, 3-fluorophenyl-
alanine, 4-fluorophenylalanine, 3-fluorophenylalanine, 3-fluorophenyl-
alanine, 2-fluorophenylalanine, 4-chlorophenylalanine, 4-nitrophenyl-
alanine, 4-aminophenylalanine, cyclohexylalanine, citrulline, 5-fluoro-
tryptophan, 5-methoxytryptophan, methionine sulfone, methionine sulfoxide
or -NH-NR~~-C(O)N(R~~)2, which are also substituted, where appropriate,
are also preferred characteristic radicals of amino acid which are employed
as the radical Ra. When amino acids which occur naturally, and also amino
acids which do not occur naturally, possess a functional group such as
amino, hydroxyl, carboxyl, mercapto, guanidyl, imidazolyl or indolyl, this
group can also be protected.
CA 02495455 2005-02-15
- 25
The N-protecting groups which are customary in peptide chemistry, for
example protecting groups of the urethane type, benzyloxycarbonyl (Z),
t-butyloxycarbonyl (Boc), 9-fluorenyloxycarbonyl (Fmoc), allyloxycarbonyl
(Aloc), or of the acid amide type, in particular formyl, acetyl or
trifluoroacetyl, and also of the alkyl type, for example benzyl, are
preferably
used as suitable protecting groups for this purpose. When an imidazole
radical is present in R8, the sulfonic acid derivative of the formula IV,
which
is employed for the sulfonamide formation, serves, for example, as the
group for protecting the imidazole nitrogen, which group can be eliminated
once again in the presence of bases such as sodium hydroxide.
The compounds of the formulae I, la and Ib are prepared as described in
patent applications WO 01/00610 and WO 01/30774. The starting
compounds for the chemical reactions are known or can be readily
prepared using methods known from the literature.
Due to the pharmacological properties, which are evident in the models
employed, of the IKB-kinase inhibitors which are used in accordance with
the invention, said inhibitors are suitable for being employed in all forms of
pain, in particular in association with pains in which inflammatory processes
play a role.
The pharmaceuticals according to the invention can be administered orally,
by inhalation, rectally or transdermally or by means of subcutaneous,
intraarticular, intraperitoneal or intravenous injection. Oral or
intraarticular
administration is preferred.
The invention also relates to a process for producing a pharmaceutical
which comprises bringing at least one compound of the formulae I or la,
together with a pharmaceutically suitable and physiologically tolerated
excipient and, where appropriate, other suitable active compounds,
additives or auxiliary substances, into a suitable form for administration.
Examples of suitable solid or galenic preparation forms are granules,
powders, sugar-coated tablets, tablets, (micro)capsules, suppositories,
syrups, juices, suspensions, emulsions, drops or injectable solutions, and
also preparations with protracted active compound release, in the
preparation of which customary auxiliary substances, such as carrier
substances, disintegrants, binders, coating agents, swelling agents,
CA 02495455 2005-02-15
26
glidants or lubricants, flavorings, sweeteners and solubilizers are used.
Frequently employed auxiliary substances which may be mentioned are
magnesium carbonate, titanium dioxide, lactose, mannitol and other
sugars, talc, milk protein, gelatin, starch, cellulose and its derivatives,
animal and vegetable oils, such as cod liver oil, sunflower oil, groundnut oil
or sesame oil, polyethylene glycol and solvents, such as sterile water and
monohydric or polyhydric alcohols, such as glycerol. The pharmaceutical
preparations are preferably produced and administered in dosage units,
with each unit containing as the active constituent a particular dose of the
compound of the formula I according to the invention. In the case of solid
dosage units, such as tablets, capsules, sugar-coated tablets or
suppositories, this dose can be up to about 1000 mg, preferably from about
50 mg to 300 mg, and, in the case of injection solutions in ampoule form,
up to about 300 mg, preferably from about 10 mg to 100 mg. Depending on
the activity of the compound according to the formulae I or la, daily doses
of from about 20 mg to 1000 mg of active compound, preferably of from
about 100 mg to 500 mg, are indicated for treating an adult patient of about
70 kg in weight. However, higher or lower daily doses may also possibly be
appropriate. The daily dose can be administered either by means of a
once-only administration in the form of a single dosage unit, or of several
smaller dosage units, or by means of the multiple administration of
subdivided doses at predetermined intervals.
As a rule, mass-spectroscopic methods (FAB-MS, ESI-MS) are used for
determining end products. Temperatures are given in degrees centigrade;
RT denotes room temperature (from 22°C to 26°C).
Abbreviations which
are used are either explained or correspond to the customary conventions.
The invention is explained in more detail below with the aid of examples.
CA 02495455 2005-02-15
27
Preparation Examples
A.1.) Synthesis of the amino acid (methyl (S)-2-amino-3-
diphenylaminopropionate (5))
OOH DEADlTPP, ~ Diphenylamine
O
O~NHZ CHaCNITHi=, a ~NHZ 100°C
_35oC O
OH
/ Hz, Pd(OH)C /
SOCIZ IMeOH MeOH
iN ~ iN \
O I / O I /
NH
~"NHZ
OMe 4 OMe
N-Benzyloxycarbonyl-L-serine-~i-lactone (2)
54.8 g (0.209 mol) of triphenylphosphine were suspended in 600 ml of
acetonitrile and the mixture was cooled down to -35°C to -45°C
while
excluding moisture. 36.4 g (0.209 mol) of diethyl azodicarboxylate were
added dropwise at this temperature within the space of 50 minutes. The
mixture was subsequently stirred at -35°C for 15 minutes. A solution of
50 g (0.209 mol) of N-benzyloxycarbonyl-L-serine (1 ) in 500 ml of
acetonitrile was then slowly added dropwise to this mixture such that the
temperature did not rise above -35°C. The mixture was then stirred at
5°C
for 12 h. In order to terminate the reaction, the reaction solution was freed
from the solvent under reduced pressure and the crude product was
purified by means of medium-pressure chromatography on silica gel.
(DCM/AcCN: 25/1 ). 20.8 g of N-benzyloxycarbonyl-L-serine-(3-lactone (2)
were obtained after the solvent had been removed; yield 45%; (see also
Org. Synth. 1991 (70) 1ff.) in fine needles.
Empirical formula C~~H11N04; M.W. = 221.2; MS (M+H) 222.1; ~H NMR
(DMSO-d6) 4.30 (m, 1 H), 4.45 (m, 1 H), 5.10 (s, 2H), 5.22 (m, 2H), 7.45 (m,
5H), 8.20 (d, J = 9.8 Hz, 1 H).
(S)-2-Benzyloxycarbonylamino-3-diphenylaminopropionic acid (3)
CA 02495455 2005-02-15
28
5.0 g (22.6 mmol) of serine lactone (2) were mixed by stirring with 20 g
(118.2 mmol) of diphenylamine, and the mixture was heated at 100°C for
2 h. The crude product was purified by means of medium-pressure
chromatography on silica gel. (DCM/methanol: 9/1, then EA/n-heptane:
4/1 ). 3.65 g (yield 42%) of pure 2-benzyloxycarbonylamino-3-
diphenylaminopropionic acid (3) were obtained after the solvent had been
removed.
Empirical formula Cz3H22N20a; M.W. = 390.44; MS (M+H) 391.2;
'H NMR (DMSO-ds) 3.85 (m, 1 H), 4.18 (m, 1 H), 4.3 (m, 1 H), 4.9 (m, 2H),
6.9 (m, 5H), 7.25 (m, 10H).
Methyl (S)-benzyloxycarbonylamino-3-diphenylaminopropionate (4)
6.5 ml (89.1 mmol) of thionyl chloride were added dropwise, at -5°C, to
75 ml of methanol and the mixture was stirred for 15 min. 3.6 g (9.22 mmol)
of 2-benzyloxycarbonylamino-3-diphenylaminopropionic acid (3), dissolved
in 75 ml of methanol, were then added and the mixture was stirred at room
temperature for a further 3 hours (h). After the solvents had been
evaporated, the residue was taken up in ethyl acetate and extracted with
sodium carbonate solution. The purification by means of flash
chromatography (n-heptane/ethyl acetate 7:3) yielded 2.76 g (50% yield) of
methyl 2-benzyloxycarbonylamino-3-diphenylaminopropionate (4).
Empirical formula C24H24N2O4; M.W. = 404.47; MS (M+H) 405.2;
'H NMR (DMSO-ds) 3.58 (s, 3H), 3.95 (m, 1 H), 4.18 (m, 1 H), 4.4 (m, 1 H),
4.95 (m, 2H), 6.9 (m, 6H), 7.3 (m, 9H), 7.85 (d, J = 9.8 Hz, 1 H).
Methyl (S)-2-amino-3-diphenylaminopropionate (5)
In order to eliminate the Z protecting group, 2.7 g (6.68 mmol) of the Z-
protected derivative (4) were dissolved in 500 ml of methanol, and 100 mg
of catalyst (10% Pd(OH)2-C) were supplied under a protective atmosphere
of nitrogen. The inert gas was subsequently displaced with a large excess
of hydrogen and the mixture was shaken for 2 h in the hydrogen
atmosphere. In order to terminate the reaction, the catalyst was filtered off
and the filtrate was concentrated. 1.65 g (yield: 91 %) of methyl 2-amino-3-
diphenylaminopropionate (5) were obtained.
Empirical formula C~gH~gN2O2; M.W. = 270.32; MS (M+H) 271.2;
'H NMR (DMSO-ds) 3.45 (s, 3H), 3.58 (m, 1 H), 3.8 (m, 1 H), 3.95 (m, 1 H),
6.9 (m, 6H), 7.3 (m, 4H).
CA 02495455 2005-02-15
29
A.2.) Synthesis of the heterocycliC parent substance (2-(2-
methylaminopyrimidin-4-yl)-1 H-indole-5-carboxylic acid (10))
Methylguanidine
Hydrochlride
O ~O \ NaOEt/EtOH,
120°C ( s.sh, reflex
\ ~ + ',.O~Ni --t w0 \ N\ N \
6 O 7 ( .O 8 \NJ\N
H 9
4-Hydrazine-
benzoic acid,
50%ige HZSO,,, H02C \ \ _
130°C .
H N \
NH
/
5 1-Dimethylamino-4,4-dimethoxypent-1-en-3-one (8)
100 g (0.76 mol) of 3,3-dimethoxy-2-butanone were stirred, at 120°C for
48 h, with 90.2 g of N,N-dimethylformamide dimethylacetal (0.76 mol). The
methanol which was formed during the reaction was continuously removed
from the reaction solution by distillation. When the solution was cooled,
10 spontaneous crystallization occurred, with this crystallization being
brought
to completion by adding a little heptane. This resulted in 128.24 g of crude
product 8 (yield 90%), which was subjected to reaction without any further
purification.
Empirical formula C9H~~N03; M.W. = 187.24; MS (M+H) 188.2;
1H NMR (DMSO-ds) 1.22 (s, 3H), 2.80 (s, 3H), 3.10 (s, 9H), 5.39 (d,
J=15Hz,1H),7.59(d,J=15Hz,1H).
[4-(1,1-Dimethoxyethyl)pyrimidin-2-ylJmethylamine (9)
1.22 g (53 mmol) of sodium were dissolved in 100 ml of absolute ethanol.
5.8 g (53 mmol) of methylguanidine hydrochloride and 10 g (53 mmol) of 1
dimethylamino-4,4-dimethoxypent-1-en-3-one (8) were added, with stirring,
to this solution, which was heated at boiling heat for 4 h. In order to
terminate the reaction, the ethanol was evaporated. The product 9, which
was obtained in this way, was used for the subsequent reaction without any
further purification. Yield 11.5 g (58 mmol, quantitative).
Empirical formula CgH~5N3O2; M.W. = 197.24; MS (M+H) 198.2;
'H NMR (DMSO-ds) 1.45 (s, 3H), 2.78 (s, 3H), 3.10 (s, 6H), 6.75 (d,
J = 3 Hz, 1 H), 7.0-7.1 (s(b), 1 H), 8.30 (d, J = 3 Hz, 1 H).
CA 02495455 2005-02-15
2-(2-Methylaminopyrimidin-4-yl)-1 H-indole-5-carboxylic acid (10)
5 g (25 mmol) of [4-(1,1-dimethoxyethyl)pyrimidin-2-yl]methylamine (9) and
3.85 g of 4-hydrazinobenzoic acid were added, while stirring, to 150 ml of
50% sulfuric acid, and the mixture was heated at 130°C for 4 h. The
5 methanol which was formed during the reaction was removed continuously
from the reaction solution by distillation. After it had been cooled down to
10°C, the reaction mixture was poured onto 200 ml of ice, and the pH
was
adjusted to about 5.5 using concentrated sodium hydroxide solution. The
precipitate of sodium sulfate and product mixture which was formed in this
10 connection was filtered off and the filter residue was extracted several
times with methanol. The combined methanol extracts were concentrated
and the product was purified by means of flash chromatography
(DCM/methanol 9:1 ). Yield: 0.76 g (11 %).
Empirical formula C~4H13N402; M.W. = 268.28; MS (M+H) 405.2; 1H NMR
15 (DMSO-ds) 2.95 (s, 3H), 6.90-7.10 (s(b), 1 H), 7.18 (d, J = 3 Hz, 1 H), 7.4
(s,
1 H), 7.58 (d, J = 4.5 Hz, 1 H), 7.80 (d, J = 4.5 Hz, 1 H), 8.30 (s, 1 H),
7.80 (d,
J = 4.5 Hz, 1 H), 8.38 (d, J = 3 Hz, 1 H), 11.85 (s, 1 H), 12.40-12.60 (s(b),
1 H).
20 A.3.) Bringing the building blocks together and synthesizing N-[(S)-2-
diphenylamino-1-(5-oxo-4,5-dihydro-[1,3,4]oxadiazol-2-yl)ethyl]-(2-(2-
methylaminopyrimidin-4-yl)-1 H-indole-5-carboxamide (13))
TOTU, DIEA, N \ ~ Hydrazine hydrate
DMF, RT t O MeOH, RT, 2 d
10 O~N I \ ~ \_.
OMe i N N-
11 N
\ \
~ Phosgenein
toluene, DCM, '
~N \ ~ RT N
O r O
O~N I \ ~ ~ N N/ ~N I \ ~ ~ N
H N~NH ~ N N~ ~ ~ N N
N O N
/ 13 /
CA 02495455 2005-02-15
31
3-Diphenylamino-2-{[2-(2-methylaminopyrimidin-4-yl)-1 H-indole-5-
carbonyl]-(S)-amino}propionic acid (11)
5.0 g (18.64 mmol) of 2-(2-methylaminopyrimidin-4-yl)-1 H-indole-5
carboxylic acid (10) were dissolved in 1.2 I of DMF, after which 7.9 g
(24.08 mmol) of TOTU and 7.9 ml (46.45 mmol) of ethyldiisopropylamine
were added consecutively. The mixture was stirred at 5°C for 20 min,
after
which 0.73 g (3.28 mmol) of (S)-2-benzyloxycarbonylamino-3-
diphenylaminopropionic acid (5) was added to the solution. After 15 h of
stirring, the mixture was concentrated under reduced pressure, after which
the residue was taken up in n-butanol and the organic phase was extracted
with a saturated solution of sodium hydrogen carbonate in order to
separate off byproducts. After the organic phase had been dried with
MgS04 and concentrated, the methyl ester of the title compound was
isolated by means of flash chromatography on silica gel (DCM:MeOH =
19:1 ). Yield: 4.3 g (98%)
Empirical formula C3pH2gNgO3; M.W. = 520.22; MS (M+H) 521.3;
'H NMR (DMSO-ds) 2.95 (s(b), 3H), 3.60 (s, 3H), 4.19-4.58 (m, 2H), 4.85
(q, 1 H), 6.90-7.10 (m, 7H), 7.18 (d, J = 3 Hz, 1 H), 7.25-7.40 (m, 5H), 7.50
(d, J = 4.5 Hz, 1 H), 7.65 (d, J = 4.5 Hz, 1 H), 8.05 (s, 1 H), 8.35 (d, J = 3
Hz,
1 H), 8.70 (d, J = 3.75 Hz, 1 H), 11.85 (s, 1 H).
N-((S)-2-Diphenylamino-1-hydrazinocarbonylethyl)-2-(2-
methylaminopyrimidin-4-yl)-1 H-indole-5-carboxamide (12)
1.0 g (1.92 mmol) of 3-diphenylamino-2-{[2-(2-methylaminopyrimidin-4-yl)
1 H-indole-5-carbonyl]-(S)-amino}propionic acid (11 ) was dissolved in 10 ml
of methanol, after which 0.48 g (9.95 mmol) of hydrazine hydrate was
added and the mixture was stirred at room temperature for 15 h. The
precipitate of the product (0.3 g) was separated off from the mother liquor
by filtration. Further hydrazone 12 (0.1 g) was isolated from the
concentrated mother liquor by flash chromatography on silica gel
(DCM:MeOH = 19:1 ). Yield: 0.4 g (40%)
Empirical formula C29H28N802; M.W. = 520.6; MS (M+H) 521.4;
'H NMR (DMSO-ds) 2.95 (s(b), 3H), 4.02-4.58 (m, 2H), 4.4 (s, 2H), 4.85 (q,
1 H), 6.90-7.10 (m, 7H), 7.18 (d, J = 3 Hz, 1 H), 7.20-7.45 (m, 5H), 7.50 (d,
J = 4.5 Hz, 1 H), 7.62 (d, J = 4.5 Hz, 1 H), 7.99 (s, 1 H), 8.25 (d, J = 3 Hz,
1 H), 8.35 (s(b), 1 H), 9.30 (s, 1 H), 11.70 (s, 1 H).
N-[(S)-2-Diphenylamino-1-(5-oxo-4,5-dihydro-[1,3,4Joxadiazol-2-yl)ethyl]-2-
(2-methylaminopyrimidin-4-yl)-1 H-indole-5-carboxamide (13)
CA 02495455 2005-02-15
32
200 mg (0.384 mmol) of N-((S)-2-diphenylamino-1-hydrazinocarbonylethyl)-
2-(2-methylaminopyrimidin-4-yl)-1 H-indole-5-carboxamide (12) were
suspended in 20 ml of methylene chloride, and a 20% solution of phosgene
in toluene (0.398 mmol) was added dropwise at 0°C and while stirring.
The
mixture was stirred at room temperature for a further 15 h and the solvent
was concentrated. The oxadiazolone 13 was subsequently isolated by flash
chromatography on silica gel (DCM:MeOH = 9:1 ). Yield: 160 mg (76%)
Empirical formula C3oH26N803; M.W. = 546.6; MS (M+H) 547.3;
1H NMR (DMSO-d6) 2.95 (s(b), 3H), 4.02-4.58 (m, 2H), 4.85 (q, 1H), 6.90
7.10 (m, 7H), 7.15 (d, J = 3 Hz, 1 H), 7.20-7.40 (m, 6H), 7.52 (d, J = 4.5 Hz,
1 H), 7.68 (d, J = 4.5 Hz, 1 H), 8.10 (s, 1 H), 8.92 (d, J = 3 Hz, 1 H), 11.78
(s,
1 H), 12.15-12.40 (s(b), 1 H).
B.) Example benzimidazole IKB-kinase inhibitor
B.1.) Synthesis of the amino acid (methyl (S)-2-amino-3-
diphenylaminopropionate (5)) as described under A.1.
B.2.) Synthesis of the heterocyclic parent substance (2-(2-
methylaminopyrimidin-4-yl)-1 H-benzimidazole-5-carboxylic acid (19))
Methylguanidine
~O ~O NaOEt/EtOH, O O
11D°C ~ 3h, reflux
'.p~ + ~O~Ni --~ \O \ N\ N \
14 O 15 O 16 1N"N
H 17
p 3,4-Diamino-
benzoic acid,
2N HzS04, Nitrobenzene, HOzC \ N
80°-3h~ N \ 150°C ~ \~~H
wN~N / H N '
NH
H 18 19 /
4-Dimethylamino-1,1-dimethoxybut-3-en-2-one (16)
300 g (307 ml, 2.54 mol) of methylglyoxal dimethylacetal were stirred, at
110°C for 4 hours (h), with 303 g (337 ml, 2.54 mol) of N,N-
dimethylformamide dimethylacetal. The methanol which was formed during
the reaction was removed continuously from the reaction solution by
distillation. After having been cooled down, the solution was extracted with
CA 02495455 2005-02-15
33
heptane and the solvents were evaporated. This resulted in 303 g of crude
product 16 (yield 70%), which was reacted without any further purification.
Empirical formula CgH~5NO3; M.W. = 173.21; MS (M+H) 174.1;
'H NMR (DMSO-d6) 2.10 (s, 1 H), 2.80 (s, 3H), 3.10 (s, 3H), 3.25 (s, 3H),
3.3 (s, 3H), 4.42 (s, 1 H), 5.19 (d(b), J = 12.8 Hz, 1 H), 7.60 (d, J = 15 Hz,
1 H).
(4-Dimethoxymethylpyrimidin-2-yl)methylamine (17)
0.33 g (14.4 mmol) of sodium was dissolved in 50 ml of absolute ethanol.
1.57 g (14.4 mmol) of methylguanidine hydrochloride and 2.48 g
(14.4 mmol) of 4-dimethylamino-1,1-dimethoxybut-3-en-2-one (16) were
added, while stirring, to the solution, which was heated at boiling heat for
3 h. In order to terminate the reaction, the ethanol was evaporated. The
resulting product 17 was used without any further purification. Yield: 2.6 g
(quantitative).
Empirical formula CaH~3N302; M.V11. = 183.21; MS (M+H) 184.1;
'H NMR (DMSO-ds) 2.78 (s, 6H), 3.10 (s, 3H), 5.02 (s, 1H), 6.62 (d,
J = 3 Hz, 1 H), 8.30 (d, J = 3 Hz, 1 H).
2-Methylaminopyrimidine-4-carbaldehyde (18)
10 g (54 mmol) of (4-dimethoxymethylpyrimidin-2-yl)methylamine (17) were
dissolved in 54 ml of 2N sulfuric acid and the solution was heated at
80°C
for 3 h while being stirred. After the reaction had cooled down, the reaction
solution was carefully brought to a pH of about 9 using solid Na2C03 and
extracted 3 times with ethanol. After the solvent had been evaporated, the
combined dried extracts yielded the title aldehyde 18 in 60% yield (4.47 g)
Empirical formula C6H~N30; M.W. = 137.12; MS (M+H) 138.2;
~H NMR (DMSO-dfi) 2.60-2.80 (s(b), 3H), 6.95 (d, J = 3 Hz, 1H), 7.40-7.60
(s(b), 1 H), 8.55 (d, J = 3 Hz, 1 H).
2-(2-Methylaminopyrimidin-4-yl)-1 H-benzimidazole-5-carboxylic acid (19)
4.3 g (31.3 mmol) of methylaminopyrimidine-4-carbaldehyde (18) and 4.8 g
(31.1 mmol) of 3,4-diaminobenzoic acid were heated at 150°C for 2 h in
300 ml of nitrobenzene. After the mixture had been cooled down to 0°C,
the
precipitate of the benzimidazole was separated off from the nitrobenzene
by filtration and the product was purified by flash chromatography
(DCM/methanol 4:1 ). Yield: 2.66 g (32%)
Empirical formula C~3H11N502~ M.W. = 269.28; MS (M+H) 270.2;
CA 02495455 2005-02-15
34
1H NMR (DMSO-ds) 2.95 (s, 3H), 7.50 (d, J = 3 Hz, 1 H), 7.75 (d,
J = 4.5 Hz, 1 H), 7.90 (d, J = 4.5 Hz, 1 H), 8.35 (s, 1 H), 8.55 (d, J = 3 Hz,
1 H), 8.70-9.05 (s(b), 1 H).
.3.) Bringing the building blocks together and synthesizing N-((S)-1-
carbamoyl-2-diphenylaminoethyl)-2-(2-methylaminopyrimidin-4-yl)-1 H-
benzimidazole-5-carboxamide (22)
TOTU, DIEA, \ I NH3/MeOH, RT,
+ DMF, RT _ !N O 15 h
20 O~N ~ N~
OMe I / N~N
2~ -~NH
I ~,
N \ I
O
N ~ N
I ~--~N
N N-
22 N H
3-Diphenylamino-2-{[2-(2-methylaminopyrimidin-4-yl)-1 H-benzimidazole-5-
carbonyl]-(S)-amino}propionic acid (21 )
2.6 g (9.6 mmol) of 2-(2-methylaminopyrimidin-4-yl)-1 H-benzimidazole-5
carboxylic acid (20) were dissolved in 300 ml of DMF, after which 3.17 g
(9.6 mmol) of TOTU and 1.6 ml (11.6 mmol) of ethyldiisopropylamine were
added consecutively. The solution was stirred at 5°C for 20 min, after
which
2.6 g (9.6 mmol) of (S)-2-benzyloxycarbonylamino-3-diphenylamino-
propionic acid (5) were added to it. After 16 h of stirring, the mixture was
concentrated under reduced pressure, after which the methyl ester 21 was
isolated by means of flash chromatography on silica gel (DCM:MeOH =
9:1 ). Yield: 1.61 g (32%)
Empirical formula C29H2~N~03; M.W. = 521.58; MS (M+H) 522.3;
1H NMR (DMSO-ds) 2.95 (s(b), 3H), 3.60 (s, 3H), 4.19-4.40 (m, 2H), 4.90
(q, 1 H), 6.90-7.10 (m, 6H), 7.25-7.35 (m, 6H), 7.40 (d, J = 4.5 Hz, 1 H),
7.60-7.80 (d(b) 1 H), 8.05-8.25 (d(b), 1 H), 8.45 (d, J = 3 Hz, 1 H), 8.90
(s(b),
1 H), 11.85 (s(b), 1 H).
CA 02495455 2005-02-15
N-((S)-1-Carbamoyl-2-diphenylaminoethyl)-2-(2-methylaminopyrimidin-4-
yl)-1 H-benzimidazole-5-carboxamide (22)
50 ml of (absolute) methanol were saturated with ammonia at 0°C. 0.5 g
(0.959 mmol) of 3-diphenylamino-2-{[2-(2-methylaminopyrimidin-4-yl)-1 H
5 benzimidazole-5-carbonyl]-(S)-amino)propionic acid (21 ) was added to this
mixture and the whole was stirred at room temperature for 24 h. After the
solvent and excess ammonia had been evaporated, the amide 22 was
isolated by flash chromatography on silica gel (DCM:MeOH = 19:1 ).
Yield: 0.43 g (89%)
10 Empirical formula C29H28Ns02; M.W. = 506.57; MS (M+H) 507.2;
'H NMR (DMSO-ds) 2.95 (s(b), 3H), 4.02-4.35 (m, 2H), 4.85 (q, 1 H), 6.80-
7.10 (m, 6H), 7.15-7.25 (m, 5H), 7.40 (d, J = 4.5 Hz, 1 H), 7.58 (s(b), 1 H),
7.68 (s(b), 1 H), 8.06-8.19 (d(b), 1 H), 8.40-8.58 (m, 2H), 13.10 (s, 1 H).
15 Pharmacological examples
IKB-kinase ELISA:
The activity of the IKB-kinase was determined using an ELISA which
comprised a biotinylated substrate peptide, which contained the amino acid
sequence in the I,cB protein from serine 32 to 36, and a specific polyclonal
20 or monoclonal antibody (e.g. from New England Biolabs, Beverly, MA,
USA, Cat.: 9240), which only bound to the phosphorylated form of the IKB
peptide. This complex was immobilized on an antibody-binding (protein A-
coated) plate and detected using a conjugate composed of a biotin-binding
protein and HRP (e.g. streptavidin-HRP). The activity was quantified with
25 the aid of a standard curve constructed using substrate phosphopeptide.
Implementation:
In order to obtain the kinase complex, 10 ml of HeLa S3 cell extract S100
were diluted with 40 ml of 50 mM HEPES, pH 7.5, brought to 40% with
30 respect to ammonium sulfate and incubated on ice for 30 minutes. The
precipitated pellet was dissolved in 5 ml of SEC buffer (50 mM HEPES,
pH 7.5, 1 mM DTT, 0.5 mM EDTA, 10 mM 2-glycerophosphate),
centrifuged at 20 000 g for 15 minutes and filtered through a 0.22 ~m filter.
The sample was loaded onto a 320 ml Superose-6 FPLC column
35 (Amersham Pharmacia Biotech AB, Uppsala, Sweden) which had been
equilibrated with SEC buffer and which was operated at 4°C with a flow
rate of 2 ml/min. The fractions which were located at the migration time of
the 670 kDa molecular weight standard were combined for the activation.
Activation was achieved by means of a 45-minute incubation with 100 nM
CA 02495455 2005-02-15
w 36
MEKK10, 250 ~M MgATP, 10 mM MgCl2, 5 mM dithiothreitol (DTT), 10 mM
2-glycerophosphate and 2.5 pM microcystin-LR at 37°C. The activated
enzyme was stored at -80°C. The test substances (2 ul), which were
dissolved in DMSO, were preincubated, at 25°C for 30 minutes, with 43
p.l
of activated enzyme (diluted 1:25 in reaction buffer 50 mM HEPES, pH 7.5,
mM MgCl2, 5 mM DTT, 10 mM (3-glycerophosphate, 2.5 pM
microcystin-LR). 5 p.l of substrate peptide (biotin-(CH2)s-
DRHDSGLDSMKD-CONH2) (200 ~.M) were then added, after which the
mixture was incubated for one hour and the reaction was stopped with
10 150 p.l of 50 mM HEPES, pH 7.5, 0.1 % BSA, 50 mM EDTA, antibody
[1:200]. 100 ~.I of the stopped reaction mixture or of a standard
phosphopeptide dilution series (biotin-(CH2)6-DRHDS[P03]GLDSMKD-
CONH2) were then transferred to a protein A plate (Pierce Chemical Co.,
Rockford, IL, USA), after which the plate was incubated for 2 hours while
being shaken. After 3 washing steps with PBS, 100 ~I of 0.5 ~g/ml
streptavidin-HRP (horseradish peroxidase) (diluted in 50 mM HEPESI0.1%
BSA) were added for 30 minutes. After 5 washing steps with PBS, 100 ~L
of TMB substrate (Kirkegaard & Perry Laboratories, Gaithersburg, MD,
USA) were added and the color development was stopped by adding
100 pL of 0.18 M sulfuric acid. The absorption was measured at 450 nm.
The standard curve was produced by linear regression corresponding to a
4-parameter dose-effect relationship. This standard curve was used to
quantify the enzyme activity or its inhibition by the test substances.
The ICSO for N-[(S)-2-diphenylamino-1-(5-oxo-4,5-dihydro[1,3,4]oxadiazol-
2-yl)ethyl]-2-(2-methylaminopyrimidin-4-yl)-1 H-indole-5-carboxamide was
0.050 pM.
The ICSO for N-((S)-1-carbamoyl-2-diphenylaminoethyl)-2-(2
methylaminopyrimidin-4-yl)-1 H-benzimidazole-5-carboxamide was
0.04.5 ~M.
Pain assay
The analgesic and antinociceptive activity of the compound N-[(S)-2-
diphenylamino-1-(5-oxo-4,5-dihydro[1,3,4]oxadiazol-2-yl)ethyl]-2-(2-
methylaminopyrimidin-4-yl)-1 H-indole-5-carboxamide, termed compound
13 in that which follows, was demonstrated in the two following models:
1St model: Zymosan-induced paw inflammation in the rat;
CA 02495455 2005-02-15
' 37
Parameter: paw withdrawal time or paw withdrawal threshold during
thermal or mechanical stimulation of the hind paw.
2"d model: Kaolin/carrageenan-induced knee joint inflammation in the rat;
Parameter: reaction of spinal neurons during pressure stimulation of
the knee.
Model 1
Experimental implementation: in short-term anesthesia using isoflurane,
1 mg of Zymosan (as a suspension in 100 ~I of PBS (phosphate-buffered
salt solution)) was injected subcutaneously into the middle of the plantar
side of one of the experimental animal's hind paws. After that, two different
behavioral tests were used to quantitatively determine the development of
a hyperalgesia.
a) Determining the paw withdrawal time during thermal stimulation
(Hargreaves test).
The experimental animal was placed in a transparent plastic chamber
having a glass floor. As soon as the experimental animal was no longer
moving, following the reconnaissance phase (about 5 min), an infrared light
source was positioned directly below the hind paw to be stimulated and
switched on. The lamp emitted focused infrared light of increasing intensity,
such that the skin temperature of the hind paw increased almost linearly.
As soon as the animal withdrew the paw, the lamp switched itself off. The
temperature of the paw at the time it is withdrawn has just become
unpleasant for the animal; this is referred to as the thermal pain threshold.
b) Determining the paw withdrawal threshold during mechanical
stimulation (von Frey test)
The experimental animal was placed in a transparent plastic chamber
whose floor consisted of wire-gauze. Punctate pressure of defined strength
was produced using calibrated nylon fibers, what are termed von Frey
hairs. The weakest pressure stimulation during which the animal withdrew
its paw determines the mechanical pain threshold.
About half an hour before, and at various times after, the Zymosan
injection, the thermal and mechanical pain thresholds were determined on
the right hind paw and on the left hind paw (see Tables 1, 2). The decrease
in the ipsilateral pain threshold, expressed in % of the contralateral pain
threshold, was then calculated (see Tables 1, 2). The degree of
hyperalgesia is directly proportional to the magnitude of this decrease.
CA 02495455 2005-02-15
38
In a control group, the Zymosan injection induced pronounced mechanical
and thermal hyperalgesia (see control data in Tabs. 1 and 2). In another
group of animals, which were under short-term isoflurane anesthesia, the
abovementioned compound 13 was injected intraperitoneally (i.p.) (in each
case 30 mg/kg in polyethylene glycol/water mixture (PEG/water 1:1 ) about
minutes before, and 2.5 and 5.5 hours after Zymosan injection. From
two hours after Zymosan injection onward, the thermal hyperalgesia was
less pronounced in these animals than it was in the control group; after the
third administration of the substance, it was no longer possible to observe
10 any side difference at all in the paw withdrawal time (Tab. 1). In
addition,
this effect still persisted for 18 hrs after the last administration of the
substance.
Compound 13 also significantly reduced the mechanical hyperalgesia. The
effect set in 1 hour after Zymosan injection and also still persisted 18 hrs
15 after the last administration of the substance (see Tab. 2).
The activity of compound 13 is very strong in both test models.
Comparative data from a study which was carried out previously show that
compound 13 reduces the thermal hyperalgesia considerably more
powerfully than does the NSAID diclofenac.
Table 1: Change in the paw withdrawal time (%)
Time (h) Mean value SD compound Mean value
after ZymosanCompound 13 13 control SD control
injection
(0)
Baseline 0.0 0.0 0.0 0.0
-0.5
0.5 -16.6 6.6 -21.4 6.3
1 -31.3 14.1 -28.8 11.6
2 -30.2 15.4 -44.8 19.1
3 -15.3 5.3 -49.2 17.9
4 -16.0 11.5 -50.6 23.0
5 -9.7 18.6 -46.6 24.8
6 5.0 2.6 -38.4 17.6
7 3.4 5.8 -29.9 22.1
24 -3.8 7.0 -46.1 18.4
CA 02495455 2005-02-15
39
Table 2:
Change in the paw withdrawal threshold (%)
Time (h) SD
after ZymosanMean value compound Mean value SD control
injection Compound 13 13 control
(0)
Baseline 0.0 0.0 0.0 0.0
-0.5
0.5 -37.4 6.6 -48.9 31.3
1 -43.1 20.5 -66.0 23.2
2 -36.0 17.8 -71.8 26.0
3 -35.1 13.1 -60.5 20.2
4 -46.7 11.9 -64.3 18.2
-40.6 14.0 -55.5 25.8
6 -33.1 23.3 -57.3 18.0
7 -44.7 21.5 -47.1 23.9
24 -9.7 26.6 -41.5 17.3
Model 2,
5 Experimental implementation: In rats which were under sodium thiopental
anesthesia, the spinal canal was opened and spinal medullary neurons
which processed the "pain impulses" from the knee-joint were identified.
Following identification, a long-term recording, in which the activity of the
nerve cell was recorded before and during the development of an acute
inflammation in the knee-joint, was carried out. For this, the responses to
non-noxious and noxious stimulation at the knee-joint were measured in a
control period before inducing the inflammation and for several hours after
inducing the inflammation.
The acute inflammation was induced by the intraarticular injection of a
suspension (about 150 ~I) of kaolin and carrageenan. In controlled
experiments, only the vehicle was applied to the spinal medullary surface in
order to represent the development of the hyperexcitability under control
conditions. As a rule, this development of hyperexcitability took place within
2 to 4 hours and was expressed in a marked increase in the responses to
non-noxious and noxious stimulation of the knee-joint (Tab.3). In the
experiments in which the abovementioned compound 13 was applied, the
substance was added (about 30 ~I of a 10 uM solution) to the spinal
medulla about 30 minutes before inducing the inflammation. The responses
of the cell to non-noxious and noxious stimulation were then subsequently
monitored as in the control experiments.
CA 02495455 2005-02-15
Comparison of the changes in the responses in the two groups shows that,
as compared with the controls, compound 13 almost completely
suppressed the development of spinal hyperexcitability (Tab.3). Taken
overall, the effect of compound 13 on the responses to noxious knee-joint
5 stimulation was more strongly expressed than was the effect of
indomethacin, as was shown by a comparison with published data from an
earlier study.
Table 3: Neuronal responses before and during knee-joint inflammation
10 (imp/15s)
Noxious stimulation at the knee-joint
Time (min)
after IVC Mean value SEM Mean valueSEM
injection Compound 13 compound 13 control control
Baseline 0.8 29.9 0 0
30-60 62.3 49.3 161.6 43.7
60-120 26.9 35 458.1 125.4
120-180 8.5 58.9 544.2 140.0
180-240 19.5 59.9 616.3 174.7
Non-noxious stimulation at the knee-joint
Time (min)
after K/C Mean value SEM Mean valueSEM
injection Compound compound 13 control control
13
Baseline 0.92 16.90 0 0
30-60 8.66 23.76 21.4 11.9
60-120 2.71 25.94 74.6 38.3
120-180 11.16 24.22 105.7 39.0
180-240 39.78 25.09 149.7 44.3
The effect of N-((S)-1-carbamoyl-2-diphenylaminoethyl)-2-(2-
15 methylaminopyrimidin-4-yl)-11i-benzimidazole-5-carboxamide, termed
compound 22 below, was also tested in model 2.
CA 02495455 2005-02-15
41
Control data: see Table 3
Table 4: Neuronal responses before and during knee-joint inflammation
(impl15s)
Noxious stimulation
at the knee-joint
Time (min) Compound 22 Compound 22
after K/C Experiment 1 Experiment 2
injection
Baseline 0 0
30-60 -109.1 -9.2
60-120 -101.1
120-180 -37.8 60
180-240 96.7
Non-noxious stimulation at the knee-joint
Time (min) Compound 22 Compound 22
after KIC Experiment 1 Experiment 2
injection
0
Baseline 0 0
30-60 -34.1 -30.6
60-120 -37.2
120-180 -32.1 50.3
180-240 68.7
The data verify the good effect of compound 22 in model 2.
CA 02495455 2005-02-15
42
3rd model: Zymosan-induced paw inflammation in the mouse;
Parameter: paw withdrawal time during thermal stimulation of the
hind paw.
Experimental implementation: In short-term anesthesia using isoflurane,
25 ~I of a suspension containing 50 mg of zymosanlml were injected into
the right hind paw of the experimental animal. The development of a
hyperalgesia was then determined quantitatively as follows:
Determining the paw withdrawal time during thermal stimulation
(Hargreave's test; see above).
The experimental animal was placed in a transparent plastic chamber
having a glass floor. As soon as the experimental animal was no longer
moving, following the reconnaissance phase (about 5 min), an infrared light
source was positioned directly below the hind paw to be stimulated and
switched on. The lamp emitted focused infrared light of increasing intensity
such that the skin temperature of the hind paw increased almost linearly.
As soon as the animal withdrew the paw, the lamp switched itself off. The
temperature of the paw at the time it is withdrawn has just become
unpleasant for the animal; this is referred to as the thermal pain threshold.
Shortly before the zymosan injection, and for from 7 to 14 days after the
injection, the thermal pain threshold was determined once daily on the right
hind paw and left hind paw. Subsequently, the integral of the area which
was formed from the curves for the paw withdrawal times of the inflamed
paw and the noninflamed paw (AUC, area between the curves, see
tables 5 and 6) was determined as a measure of the hyperalgesia. The
larger this value is, the more pronounced is the hyperalgesia, and the
smaller the value is in animals which are being given the substance, the
greater is the success of the therapy.
In a 7-day study, the zymosan injection induced pronounced thermal
hyperalgesia in a control group (see vehicle, tab. 5). In the other groups,
the substance was administered for the first time one day after the
zymosan injection, after marked thermal hyperalgesia had already
developed. Compound 13 was then administered orally twice daily for 7
days, in each case at the rate of 25 or 75 mglkg in HEC/lipofundin (1%
HEC in lipofundin). Analysis of the paw withdrawal times during the entire
period of the study (7 days) showed that, when the substance was
30-60 -109.1
CA 02495455 2005-02-15
43
administered, the AUC decreased in a dose-dependent manner. At single
doses of from 8.3 mg/kg and upwards, a significant therapeutic effect was
achieved as compared with the vehicle group (tab. 5). Compound 13
exhibits very strong activity in the test model. A very high dose of
paracetamol was likewise administered twice daily to another group of
animals which was taken through the experiment in parallel. Compound 13
reduced the thermal hyperalgesia to a greater extent than did paracetamol
(tab. 5).
Table 5 Thermal hyperalgesia during the seven days following zymosan
infection
AUC Standard Number Statistical
mean valueerror of difference
[measure of the arithmeticanimals as
of mean (SEM) per compared
h eral group with
esia the vehicle
Vehicle 45.1 1.5 8
Paracetamol, 200 24.6 4.1 8 es
m /k
Com ound 13, 2.8 40.4 2.4 8 no
m /k
Com ound 13, 8.3 32.3 2.2 8 es
m /k
Com ound 13, 25 19.4 2.9 8 es
m /k
Com ound 13, 75 17.4 2.6 8 es
m /k
In another study carried out on mice, the activity of compound 13 was
compared with that of the specific COX-2 inhibitor Celecoxib. The scheme
for zymosan injection and dosing was identical to that in the previously
described study. The only difference was that this additional study ran for
14 days.
Once again, compound 13 was able to reduce thermal hyperalgesia in a
dose-dependent manner (tab. 6). In the experiment, compound 13 and
Celecoxib had equally strong effects at the high dosage (tab. 6).
CA 02495455 2005-02-15
44
Table 6 Thermal hyperalgesia during the 14 days following zymosan
infection
AUC Standard Number Statistical
mean valueerror of difference
[measure of the arithmeticanimals as
of mean (SEM) per compared
h peral group with
esia the vehicle
Vehicle 90.0 5.1 8
Celecoxib, 8.3 79.9 5.9 5 no
m /k
Celecoxib, 25 51.5 3.7 9 no
m /k
Com ound 13, 8.3 64.5 5.0 5 es
m /k
Com ound 13, 25 47.6 4.4 9 es
m /k
4th model: Zymosan-induced paw inflammation in the mouse;
Parameter: spontaneous running performance in a running wheel.
In the cage in which it is kept, the experimental animal has access to a
running wheel, the revolutions of which are recorded electronically. During
the night hours, the C57IB6 mice use the running wheel voluntarily and,
after a one-week phase of acclimatization, cover on average
4 100 meters/night. After zymosan has been injected, the distance run
each night is reduced. This reduction in running performance is a valid
parameter for a restriction in function which is due to inflammation pain.
Experimental implementation: After an acclimatization phase of one
week, the distance run/24 hours was measured in order to determine the
base line. 25 NI of a suspension containing 50 mg of zymosan/ml were then
injected into the right hind paw of the experimental animal during short-term
anesthesia using isoflurane. The distance run/24 hours was then
determined during the following seven days. In the analysis, the area under
the curve for the values for the distance run was determined (AUC, tab. 7):
the lower the AUC, the lower was the running performance during the week
following injection of the zymosan. Compound 13 was administered twice
daily for 7 days, with the dose in each case being 25 or 75 mglkg in
HECllipofundin (1% HEC in lipofundin). The substance was administered
for the first time on day 1 after injection of the zymosan.
In one study, the effect of compound 13 on running performance following
zymosan injection was compared with that of paracetamol. A dose-
dependent increase in the distance run, which was significant as compared
with the vehicle group, was found in the case of both the higher doses
(tab. 7). By contrast, no improvement as compared with the vehicle group
CA 02495455 2005-02-15
was achieved when paracetamol was used at an extremely high dose (also
2 x daily) (tab. 7).
Table 7 Running wheel activity during the seven days following zymosan
5 in'ection
AUC Standard Number Statistical
mean valueerror of difference
of the arithmeticanimals as
mean (SEM) per compared
group with
the vehicle
Vehicle 108.8 12.5 8
Paracetamol, 200 187.2 42.7 8 no
m /k
Com ound 13, 2.8 131.1 23.3 8 no
m /k
Com ound 13, 8.3 142.1 29.1 8 no
m lk
Com ound 13, 25 216.7 58.5 8 es
m /k
Compound 13, 75 251.7 41.9 8 es
m /k
