Note: Descriptions are shown in the official language in which they were submitted.
CA 02512183 2005-06-29
WO 20041060883 PCTIEP20031014612
1
PYRIMIDINE-4, 6-DICARBOXYLIC ACID DIAMIDES FOR USE AS
SELECTIVE MMP 13 INHIBITORS
The invention relates to the use of pyrimidine-4,6-dicarboxylic acid
diamides for selectively inhibiting collagenase (MMP 13). The pyrimidine-
4,6-dicarboxylic acid diamides can therefore be used for treating
degenerative joint diseases.
In diseases such as osteoarthritis and rheumatism, destruction of the joint
takes place, with this destruction being caused, in particular, by the
proteolytic breakdown of collagen due ,to collagenases. Collagenases
belong to the metalloproteinase (MP) or matrix metalloproteinase (MMP)
superfamily. Under physiological conditions, MMPs cleave collagen,
laminin, proteoglycans, elastin or gelatin and therefore play an important
role in bone and connective tissue. A large number of different inhibitors of
the MMPs and/or collagenases have been disclosed (EP 0 606 046;
WO 94/28889). Known MMP inhibitors frequently suffer from the
disadvantage of lacking the specificity involved in inhibiting only one class
of MMPs. As a result, most MMP inhibitors inhibit several MMPs
simultaneously because the structure of the catalytic domain in the MMPs
is similar. As a consequence, the inhibitors have the undesirable property
of acting on many enzymes including those which have a vital function
(Massova l., et al., The FASEB Journal (1998) 12, 1075-1095).
It is known that pyrimidine-4,6-dicarboxylic acid diamides and 2,4-
substituted pyridine N-oxides inhibit the enzymes proline hydroxylase and
lysine hydroxylase and thereby bring about an inhibition of collagen
biosynthesis by exerting an influence on the collagen-specific hydroxylation
reaction (EP 0418797; EP 0463592). This inhibition of collagen
biosynthesis results in the formation of a nonfunctional, under-hydroxylated
collagen molecule which the cells can only release into the extracellular
space in small quantity. fn addition, the under-hydroxylated collagen cannot
be incorporated into the collagen matrix and is very readily degraded
proteolytically. As a consequence of these effects, the overall quantity of
collagen which is deposited extracellularly decreases. It is known from
patent applications WO 02/064571 and WO 02/064080 that certain
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2
pyridine-2,4-dicarboxylic acid diamides and pyrimidine-4,6-dicarboxylic acid
diamides can be allosteric inhibitors of MMP 13.
WO 02/064571 describes pyrimidine-4,6-dicarboxylic acid and pyrimidine-
4,6-dicarboxylic acid derivatives and their selective inhibition of MMP 13.
EP 0418797 also describes pyrimidine-4,6-dicarboxylic acid derivatives and
their inhibitory effect on proline hydroxylase. It was possible to confirm
this
inhibition by pyrimidine-4,6-dicarboxylic acid in in-house experiments. The
compounds described in WO 02/064571 and EP 0418797 therefore suffer
from the disadvantage that, as a result of proline hydroxylase being
inhibited, collagen biosynthesis is also inhibited and a nonfunctional, under-
hydroxylated collagen molecule is formed, with the cells only being able to
release this molecule into the extracellular space in small quantities.
The object of the present invention is therefore to find compounds which,
while selectively inhibiting MMP 13, do not inhibit proline hydroxylase and
are therefore better suited for treating degenerative joint diseases more
efficiently and more specifically.
It has now been found that the compounds employed in accordance with
the invention are powerful inhibitors of matrix metalloproteinase 13
whereas the compounds employed in accordance with the invention have
essentially no activity on the MMPs 3 and 8 and do not inhibit proline
hydroxylase.
The invention therefore relates to the use of compounds of the formula I
,o 0
R1 ~N ~ R6
U)
I
~N
R5
R4
and/or all stereoisomeric forms of the compound of the formula I and/or
mixtures of these forms in any ratio, and/or a physiologically tolerated salt
of the compound of the formula I
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3
for producing a pharmaceutical for the prophylaxis and therapy of diseases
whose course involves an increased activity of matrix metallop~oteinase 13,
where
R1 is hydrogen atom or -(C~-Cg)-alkyl,
R2 is -(C~-Cg)-alkyl, where alkyl is substituted, once, twice or three
times, by
1. -C(O)-O-R8, in which R8 is
1.1 ) hydrogen atom, or
1.2) -(C~-Cg)-alkyl,
2. -(C~-Cg)-alkyl-O-R8, in which R8 is
2.1 ) hydrogen atom, or
2.2) -(C~-Cg)-alkyl,
3. -(Cg-C~4)-aryl, in which aryl is substituted, once, twice
or three times, independently of each other, by
3.1 ) -(C2-Cg)-alkyl-C(O)-O-R8, in which R8 is
3.11 ) hydrogen atom, or
3.12) -(C~-Cg)-alkyl,
3.2) -O-(C~-Cg)-alkyl-C(O)-O-R8, in which R8
is
3.21) hydrogen atom, or
3.22) -(C~-Cg)-alkyl,
3.3) -N(R14)-(R15), in which R14 and R15
form, together with the nitrogen atom to
which they are bonded, a 5-, 6- or 7
membered saturated ring where a
heteroatom from the series oxygen,
sulfur and nitrogen can also replace one
or two further carbon atoms and, in the
case of nitrogen, the nitrogen atoms can,
independently of each other, be
unsubstituted or substituted by (C~-Cg)-
alkyl,
3.4) -(CHZ)k-N(R9)-(R10), in which k is 2, 3, 4
or 5 and R9 and R10 are identical or
different and are, independently of each
other,
3.41 ) hydrogen atom, or
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3.42) -(C~-Cg)-alkyl, or
R9 and R10 form, together with the
nitrogen atom to which they are bonded,
a 5-, 6- or 7-membered saturated ring
where a heteroatom from the series
oxygen, sulfur and nitrogen can also
replace one or two further carbon atoms
and, in the case of nitrogen, the nitrogen
atoms can, independently of each other,
be unsubstituted or substituted by
(C~-Cg)-alkyl,
3.5) -O-(C2-Cg)-alkyl-N(R9)-R10, where R9
and R10 have the abovementioned
meaning,
3.6) -N(R8)-C(O)-(C~-Cg)-alkyl, in which alkyl
is unsubstituted or substituted, once,
twice or three times, by
~ 3.61 ) halogen,
3.62) cyano,
3.63) vitro,
3.64) hydroxyl,
3.65) amino,
3.66) -C(O)-O-(C~-Cg)-alkyl, or
3.67) -C(O)-OH,
4. Het, where Het is a saturated or unsaturated
monocyclic or bicyclic, 3- to 10-membered heterocyclic
ring system which contains 1, 2 or 3 identical or
different ring heteroatoms from the series nitrogen,
oxygen and sulfur and is unsubstituted or substituted,
once or more than once, by
4.1 ) halogen,
4.2) cyano,
4.3) vitro,
4.4) hydroxyl,
4.5) amino,
4.6) -C(O)-O-(C~-Cg)-alkyl,
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4.7) -C(O)-OH,
4.8) -(C~-Cg)-alkyl, where alkyl is
unsubstituted or substituted, once, twice
or three times, by halogen,
5 4.9) -O-(C~-C6)-alkyl, where alkyl is
unsubstituted or substituted, once, twice
or three times, by halogen,
4.10) pyridyl, or
4.11 ) phenyl, where phenyl is unsubstituted
or
substituted, once or more than once and
independently of each other, by a radical
from the series halogen, (C~-C6)-alkoxy
and (C~-Cg)-alkyl,
and
R3, R4, R5, R6
and R7 are identical
or different and
are, independently
of
each other,
1. hydrogen atom,
2. halogen,
3. -(C~-Cg)-alkyl, in which alkyl is unsubstituted
or
substituted, once, twice or three times,
by halogen,
4. -O-(C~-C6)-alkyl, in which alkyl is unsubstituted
or
substituted, once, twice or three times,
by halogen, or
5. -S-(C~-C6)-alkyl.
The invention also relates to the use of the compounds of the formula I,
where
R1 is hydrogen atom or -(C~-C4)-alkyl,
R2 is -(C~-C4)-alkyl, where alkyl is substituted, once, twice or three
times, by
1. -C(O)-O-R8, in which R8 is
1.1 ) hydrogen atom, or
1.2) -(C~-C4)-alkyl,
2. -(C~-C4)-alkyl-O-R8, in which R8 is
2.1 ) hydrogen atom, or
2.2) -(C~-C4)-alkyl,
3. phenyl, in which phenyl is substituted, once, twice or
three times, independently of each other, by
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3.1 ) -(C2-C4)-alkyl-C(O)-O-R8, in which R8 is
3.11 ) hydrogen atom, or
3.12) -(C~-C4)-alkyl,
3.2) -O-(C1-C4)-alkyl-C(O)-O-R8, in which R8
is
3.21 ) hydrogen atom, or
3.22) -(C~-C4)-alkyl,
3.3) -N(R14)-(R15), in which R14 and R15
form, together with the nitrogen atom to
which they are bonded, a 5-, 6- or 7
membered saturated ring where a
heteroatom from the series oxygen,
sulfur and nitrogen can also replace one
or two further carbon atoms and, in the
case of nitrogen, the nitrogen atoms can,
independently of each other, be
unsubstituted or substituted by (C~-C4)-
alkyl,
3.4) -(CH2)k-N(R9)-(R10), in which k is 2, 3 or
4 and R9 and R10 are identical or
different and are, independently of each
other,
3.41) hydrogen atom, or
3.42) -(C~-C4)-alkyl, or
R9 and R10 form, together with the
nitrogen atom to which they are bonded,
a 5-, 6- or 7-membered saturated ring
where a heteroatom from the series
oxygen, sulfur and nitrogen can also
replace one or two further carbon atoms
and, in the case of nitrogen, the nitrogen
atoms can, independently of each other,
be unsubstituted or substituted by
(C~-C4)-alkyl,
3.5) -O-(C2-C4)-alkyl-N(R9)-R10, where R9
and R10 have the abovementioned
meaning,
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3.6) -N(H)-C(O)-(C1-C4)-alkyl, in which alkyl
is unsubstituted or substituted, once,
twice or three times, by
3.61 ) halogen,
3.62) cyano,
3.63) vitro,
3.64) hydroxyl,
3.65) amino,
3.66) -C(O)-O-(C1-Cg)-alkyl, or
3.67) -C(O)-OH,
4. Het, where Het is azepine, azetidine, aziridine,
benzimidazole, benzo[1,4]dioxin, 1,3-benzodioxole,
benzofuran, 4H-benzo[1,4)oxazine, benzoxazole,
benzothiazole, benzothiophene, quinazoline, quinoline,
quinoxaline, chroman, cinnoline, oxirane,
1,2-diazepine, 1,3-diazepine, 1,4-diazepine, 1,4-dioxin,
dioxole, furan, imidazole, indazole, indole, isoquinoline,
isochroman, isoindole, isoxazole, isothiazole,
1,2-oxazine, 1,3-oxazine, 1,4-oxazine, oxazole,
phthalazine, piperidine, pyran, pyrazine, pyrazole,
pyridazine, pyridine, pyrimidine, pyridoimidazole,
pyridopyridine, pyridopyrimidine, pyrrol, tetrazole,
1,2-thiazine, 1,3-thiazine, 1,4-thiazine, thiazole,
thiophene, thiopyran, 1,2,3-triazine, 1,2,4-triazine,
1,3,5-triazine, 1,2,3-triazole or 1,2,4-triazole and
Het is
unsubstituted or substituted, once, twice or three
times, independently of each other, by
4.1 ) halogen,
4.2) amino,
4.3) -C(O)-O-(C~-C4)-alkyl,
4.4) -C(O)-OH,
4.5) -(C~-Cg)-alkyl, where alkyl is
unsubstituted or substituted, once, twice
or three times, by halogen,
4.6) -O-(C~-Cg)-alkyl, where alkyl is
unsubstituted or substituted, once, twice
or three times, by halogen,
4.7) pyridyl, or
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4.8) phenyl, where phenyl is unsubstituted or
substituted, once or more than once and
independently of each other, by a radical
from the series halogen, -(C1-C4)-alkoxy
and -(C1-C4)-alkyl, and
R3, R4, R5, R6 and R7 are identical or different and are
1. hydrogen atom,
2. halogen,
3. -(C~-Cg)-alkyl, in which alkyl is unsubstituted or
substituted, once, twice or three times, by halogen, or
4. -O-(C~-Cg)-alkyl, in which alkyl is unsubstituted or
substituted, once, twice or three times, by halogen.
The invention also relates to the use of the compounds of the formula I,
where
R1 is hydrogen atom,
R2 is -(C1-C2)-alkyl, where alkyl is substituted, once, twice or three
times, by
1. phenyl, in which phenyl is substituted, once, twice or
three times, independently of each other, by
1.1 ) -(C2-C4)-alkyl-C(O)-O-R8, in which R8 is
1.11) hydrogen atom, or
1.12) -(C~-C4)-alkyl,
1.2) -O-(C~-C4)-alkyl-C(O)-O-R8, in which R8
is
1.21 ) hydrogen atom, or
1.22) -(C~-C4)-alkyl,
1.3) -N(R14)-(R15), in which R14 and R15
form, together with the nitrogen atom to
which they are bonded, a radical which
can be derived from imidazolidine,
isothiazolidine, isoxazolidine, morpholine,
piperazine, piperidine, pyrazine,
pyrazolidine, pyrrolidine, tetrazine or
thiomorpholine and, in the case of
nitrogen, the nitrogen atoms can,
independently of each other, be
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9
unsubstituted or substituted by -(C~-C4)-
alkyl,
1.4) -(CH2)k-N(R9)-(R10), in which k is 2, 3 or
4 and R9 and R10 are identical or
different and are, independently of each
other,
1.41) hydrogen atom, or
1.42) -(C~-C4)-alkyl, or
R9 and R10 form, together with the nitrogen
atom to which they are bonded, a radical
which can be derived from imidazolidine,
isothiazolidine, isoxazolidine, morpholine,
piperazine, piperidine, pyrazine,
pyrazolidine, pyrrolidine, tetrazine or
thiomorpholine and, in the case of
nitrogen, the nitrogen atoms can,
independently of each other, be
unsubstituted or substituted by -(C~-C4)-
alkyl,
1.5) -O-(C2-C4)-alkyl-N(R9)-R10, where R9
and R10 have the abovementioned
meaning,
1.6) -N(H)-C(O)-(C~-C4)-alkyl, in which alkyl
is unsubstituted or substituted, once,
twice or three times, by
1.61 ) halogen,
1.62) -C(O)-O-(C~-C4)-alkyl, or
1.63) -C(O)-OH,
2. Het, where Het is furan, imidazole, isothiazole,
isoxazole, oxazole, pyrazole, pyridazine, pyridine,
pyrimidine, pyrrole, thiazole, thiophene, 1,2,3-triazole
or 1,2,4-triazole, and Het is unsubstituted or
substituted, once, twice or three times, independently
of each other, by
2.1 ) halogen,
2.2) amino,
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2.3) -C(O)-O-(C~-C4)-alkyl,
2.4) -C(O)-OH,
2.5) -(C~-C4)-alkyl, where alkyl is
unsubstituted or substituted, once, twice
5 or three times, by halogen,
2.6) -O-(C~-C4)-alkyl, where alkyl is
unsubstituted or substituted, once, twice
or three times, by halogen,
2.7) pyridyl,
10 2.8) phenyl, where phenyl is unsubstituted or
substituted, once or more than once and
independently of each other, by a radical
from the series halogen, -(C~-C4)-alkoxy
and -(C~-C4)-alkyl, and
R3, R4, R5, R6
and R7 are identical
or different
and are
1. hydrogen atom,
2. halogen,
3. methyl,
~ 4. trifluoromethyl,
5. methoxy, or
6, trifluoromethoxy.
The invention also relates to the compound of the formula I
O O R7
R1 ~R \ H \ R6 ~ p
N~\ j N
R3 Y ~R5
and/or all stereoisomeric forms of the compound of the formula I and/or
mixtures of these forms in any ratio, and/or a physiologically tolerated salt-
of the compound of the formula l, where
R1 is hydrogen atom or -(C~-Cg)-alkyl,
R2 is -(C~-Cg)-alkyl, where alkyl is substituted, once, twice or three
times, by
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1. -C(O)-0-R8, in which R8 is
1.1 ) hydrogen atom, or
1.2) -(C~-Cg)-alkyl,
2. -(C~-Cg)-alkyl-O-R8, in which R8 is
2.1 ) hydrogen atom, or
2.2) -(C~-Cg)-alkyl,
3. -(Cg-C~4)-aryl, in which aryl is substituted, once, twice
or three times, independently of each other, by
3.1 ) -(C2-Cg)-alkyl-C(O)-O-R8, in which R8 is
3.11 ) hydrogen atom, or
3.12) -(C~-Cg)-alkyl,
3.2) -O-(C~-Cg)-alkyl-C(O)-O-R8, in which R8
is
3.21 ) hydrogen atom, or
3.22) -(C~-Cg)-alkyl,
3.3) -N(R14)-(R15), in which R14 and R15
form, together with the nitrogen atom to
which they are bonded, a 5-, 6- or 7-
membered saturated ring where a
heteroatom from the series oxygen,
sulfur and nitrogen can also replace one
or two further carbon atoms and, in the
case of nitrogen, the nitrogen atoms can,
independently of each other, be
unsubstituted or substituted by (C~-Cg)-
alkyl,
3.4) -(CH2)k-N(R9)-(R10), in which k is 2, 3, 4
or 5 and R9 and R10 are identical or
different and are, independently of each
other,
3.41 ) hydrogen, or
3.42) -(C~-Cg)-alkyl, or
R9 and R10 form, together with the
nitrogen atom to which they are bonded,
a 5-, 6- or 7-membered saturated ring
where a heteroatom from the series
oxygen, sulfur and nitrogen can also
CA 02512183 2005-06-29
12
replace one or two further carbon atoms
and, in the case of nitrogen, the nitrogen
atoms can, independently of each other,
be unsubstituted or substituted by
(Cq-C6)-alkyl,
3.5) -O-(C2-Cg)-alkyl-N(R9)-R10, where R9
and R10 have the abovementioned
meaning,
3.6) -N(R8)-C(O)-(C1-Cg)-alkyl, in which alkyl
is unsubstituted or substituted, once,
twice or three times, by
3.61 ) halogen,
3.62) cyano,
3.63) vitro,
3.64) hydroxyl,
3.65) amino,
3.67) -C(O)-O-(C1-Cg)-alkyl, or
3.68) -C(O)-OH,
4. Het, where Het is a saturated or unsaturated
monocyclic or bicyclic, 3- to 10-membered heterocyclic
ring system which contains 1, 2 or 3 identical or
different ring heteroatoms from the series nitrogen,
oxygen and sulfur and
is unsubstituted or substituted,
once or more than once, by
4.1 ) halogen,
4.2) cyano,
4.3) vitro,
4.4) hydroxyl,
4.5) amino,
4.6) -C(O)-O-(C~-Cg)-alkyl,
4.7) -C(O)-OH,
4.8) -(C~-Cg)-alkyl, where alkyl
is
unsubstituted or substituted,once,
twice
or three times, by halogen,
4.9) -O-(C~-Cs)-alkyl, wherealkyl
is
unsubstituted or substituted,once,
twice
or three times, by halogen,
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4.10) pyridyl, or
4.11 ) phenyl, where phenyl is unsubstituted or
substituted, once or more than once and
independently of each other, by a radical
from the series halogen, (C~-Cg)-alkoxy
and (C~-Cg)-alkyl, and
R3, R4, R5, R6 and R7 are identical or different and are, independently of
each other,
1. hydrogen atom,
2. halogen,
3. -(C~-Cg)-alkyl, in which alkyl is unsubstituted or
substituted, once, twice or three times, by halogen,
4. -O-(C~-Cg)-alkyl, in which alkyl is unsubstituted or
substituted, once, twice or three times, by halogen, or
5. -S-(C,-C6)-alkyl.
The invention also relates to the compound of the formula I, where
R1 is hydrogen atom or-(C~-C4)-alkyl,
R2 is -(C~-C4)-alkyl, where alkyl is substituted, once, twice or three
times, by
1. -C(O)-O-R8, in which R8 is
1.1 ) hydrogen atom, or
1.2) -(C~-C4)-alkyl,
2. -(C~-C4)-alkyl-O-R8, in which R8 is
2.1 ) hydrogen atom, or
2.2) -(C~-C4)-alkyl,
3. phenyl, in which phenyl is substituted, once, twice or
three times, independently of each other, by
3.1) -(C2-C4)-alkyl-C(O)-O-R8, in which R8 is
3.11 ) hydrogen atom, or
3.12) -(C~-C4)-alkyl,
3.2) -O-(C~-C4)-alkyl-C(O)-O-R8, in which R8
is
3.21) hydrogen atom, or
3.22) -(C~-C4)-alkyl,
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14
3.3) -N(R14)-(R15), in which R14 and R15
form, together with the nitrogen atom to
which they are bonded, a 5-, 6- or 7-
membered saturated ring which can be
derived from imidazolidine,
isothiazolidine, isoxazolidine, morpholine,
piperazine, piperidine, pyrazine,
pyrazolidine, pyrrolidine, tetrazine or
thiomorpholine, where the nitrogen atoms
can, independently of each other, be
unsubstituted or substituted by -(C~-C4)-
alkyl,
3.4) -(CH2)k-N(R9)-(R10), in which k is 2, 3 or
4 and R9 and R10 are identical or
different and are, independently of each
other,
3.41 ) hydrogen atom, or
3.42) -(C~-C4)-alkyl, or
R9 and R10 form, together with the
nitrogen atom to which they are bonded,
a 5-, 6- or 7-membered saturated ring
which can be derived from imidazolidine,
isothiazolidine, isoxazolidine, morpholine,
piperazine, piperidine, pyrazine,
pyrazolidine, pyrrolidine, tetrazine or
thiomorpholine, where the nitrogen atoms
can, independently of each other, be
unsubstituted or substituted by (C~-C4)-
alkyl,
3.5) -O-(C2-C4)-alkyl-N(R9)-R10, where R9
and R10 have the abovementioned
meaning,
3.6) -N(H)-C(O)-(C~-C4)-alkyl, in which alkyl
is unsubstituted or substituted, once,
twice or three times, by
3.61 ) halogen,
3.62) cyano,
CA 02512183 2005-06-29
3.63) vitro,
3.64) hydroxyl,
3.65) amino,
3.67) -C(O)-O-(C~-Cg)-alkyl, or
5 3.68) -C(O)-OH,
4. Het, where Het is azepine, azetidine, aziridine,
benzimidazole, benzo[1,4]dioxin, 1,3-benzodioxole,
benzofuran, 4H-benzo[1,4]oxazine, benzoxazole,
benzothiazole, benzothiophene, quinazoline, quinoline,
10 quinoxaline, chroman, cinnoline, oxirane,
1,2-diazepine, 1,3-diazepine, 1,4-diazepine, 1,4-dioxin,
dioxole, furan, imidazole, indazole, indole, isoquinoline,
isochroman, isoindole, isoxazole, isothiazole,
1,2-oxazine, 1,3-oxazine, 1,4-oxazine, oxazole,
15 phthalazine, piperidine, pyran, pyrazine, pyrazole,
pyridazine, pyridine, pyrimidine, pyridoimidazole,
pyridopyridine, pyridopyrimidine, pyrrole, tetrazole,
1,2-thiazine, 1,3-thiazine, 1,4-thiazine, thiazole,
thiophene, thiopyran, 1,2,3-triazine, 1,2,4-triazine,
1,3,5-triazine, 1,2,3-triazole or 1,2,4-triazole,
and Het is
unsubstituted or substituted, once, twice or three
times, independently of each other, by
4.1 ) halogen,
4.2) amino,
4.3) -C(O)-O-(C~-C4)-alkyl,
4.4) -C(O)-OH, ,
4.5) -(C~-Cg)-alkyl, where alkyl is
unsubstituted or substituted, once, twice
or three times, by halogen,
4.6) -O-(C~-Cg)-alkyl, where alkyl is
unsubstituted or substituted, once, twice
or three times, by halogen,
4.7) pyridyl, or
4.8) phenyl, where phenyl is unsubstituted or
substituted, once or more than once and
independently of each other, by a radical
from the series halogen, -(C~-C4)-alkoxy
and -(C~-C4)-alkyl, and
CA 02512183 2005-06-29
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R3, R4, R5, R6 and R7 are identical or different and are
1. hydrogen atom,
2. halogen,
3. -(C~-Cg)-alkyl, in which alkyl is unsubstituted or
substituted, once, twice or three times, by halogen, or
4. -O-(C~-Cg)-alkyl, in which alkyl is unsubstituted or
substituted, once, twice or three times, by halogen.
The invention also relates to the compound of the formula I, where
15
R1 is hydrogen atom,
R2 is -(C~-C2)-alkyl, where alkyl is substituted, once, twice or three
times, by
1. phenyl, in which phenyl is substituted, once, twice or
three times, independently of each other, by
1.1 ) -(C2-C4)-alkyl-C(O)-O-R8, in which R8 is
1.11 ) hydrogen atom, or
1.12) -(C~-C4)-alkyl,
1.2) -O-(C~-C4)-alkyl-C(O)-O-R8, in which R8
is
1.21 ) hydrogen atom, or
1.22) -(C~-C4)-alkyl,
1.3) -N(R14)-(R15), in which R14 and R15
form, together with the nitrogen atom to
which they are bonded, a radical which
can be derived from imidazolidine,
isothiazolidine, isoxazolidine, morpholine,
piperazine, piperidine, pyrazine,
pyrazolidine, pyrrolidine, tetrazine or
thiomorpholine and, in the case of
nitrogen, the nitrogen atoms can,
independently of each other, be
unsubstituted or substituted by (C1-C4)-
alkyl,
1.4) -(CH2)k-N(R9)-(R10), in which k is 2, 3 or
4 and R9 and R10 are identical or
different and are, independently of each
other,
1.41 ) hydrogen atom, or
CA 02512183 2005-06-29
17 ,
1.42) -(C1-C4)-alkyl, or
R9 and R10 form, together with the
nitrogen atom to which they are bonded,
a radical which can be derived from
imidazolidine, isothiazolidine,
isoxazolidine, morpholine, piperazine,
piperidine, pyrazine, pyrazolidine,
pyrrolidine; tetrazine or thiomorpholine
and, in the case of nitrogen, the nitrogen
atoms can, independently of each other,
be unsubstituted or substituted by
-(C1-C4)-alkyl,
1.5) -O-(C2-C4)-alkyl-N(R9)-R10, where R9
and R10 have the abovementioned
meaning,
1.6) -N(H)-C(O)-(C1-C4)-alkyl, in which alkyl
is unsubstituted or substituted, once,
twice or three times, by
1.61 ) halogen,
1.62) -C(O)-O-(C1-C4)-alkyl, or
1.63) -C(O)-OH,
2. Het, where Het is furan, imidazole, isothiazole,
isoxazole, oxazole, pyrazole, pyridazine,
pyridine,
pyrimidine, pyrrole, thiazole, thiophene,
1,2,3-triazole
or 1,2,4-tr iazole, and Het is unsubstituted
or
substituted, once, twice or three times, independently
of each other, by
2.1 ) halogen,
2.2) amino,
2.3) -C(O)-O-(C1-C4)-alkyl,
2.4) -C(O)-OH,
2.5) -(C1-C4)-alkyl, where alkyl is
unsubstituted or substituted,
once, twice
or three times, by halogen,
2.6) -O-(C1-C4)-alkyl, where alkyl
is
unsubstituted or substituted,
once, twice
or three times, by halogen,
CA 02512183 2005-06-29
18
2.7) pyridyl,
2.8) phenyl, where phenyl is unsubstituted or
substituted, once or more than once and
independently of each other, by a radical
from the series halogen, -(C~-C4)-alkoxy
and -(C~-C4)-alkyl, and
R3, R4, R5, R6 and R7 are identical or different and are
1. hydrogen atom,
2. halogen,
3. methyl,
4. trifluaromethyl,
5. methoxy, or
6. trifluoromethoxy.
The term "halogen" is understood as meaning fluorine, chlorine, bromine or
iodine.
The term "-(C~-Cg)-alkyl" is understood as meaning, in the widest possible
sense, hydrocarbon radicals containing 1, 2, 3, 4, 5 or 6 carbon atoms and
whose carbon chain is straight or branched or which consist of cyclic
hydrocarbon groups or of combinations of linear and cyclic groups. For
example, linear and branched hydrocarbon radicals can be methyl, ethyl,
propyl, i-propyl, butyl, tertiary butyl, pentyl or hexyl, while cyclic groups
can
be cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, and a combination of
linear and cyclic radicals can be cyclopropylmethyl, cyclobutylmethyl or
cyclopentylmethyl.
The phrase "R4 and R5 or R5 and R6 form, together with the carbon atoms
to which they are in each case bonded, a 5- or 6-membered ring which is
aromatic or saturated and contains zero, one or two heteroatoms from the
series oxygen, nitrogen or sulfur" is understood as meaning radicals which
can be derived, for example, from dioxolane, pyrrole, pyrrolidine, pyridine,
piperidine, dioxane, tetrahydropyridine, pyrazole, imidazole, pyrazoline,
imidazoline, pyrazolidine, imidazolidine, pyridazine, pyrimidine, pyrazine,
piperazine, pyran, furan, dihydrofuran, tetrahydrofuran, oxazole, isoxazole,
2-isoxazoline, isoxazolidine, morpholine, oxothiolane, thiopyran, thiazole,
isothiazole, 2-isothiazoline, isothiazolidine or thiomorpholine.
The term "-(Cg-C~4)-aryl" is understood as meaning aromatic carbon
radicals having from 6 to 14 carbon atoms in the ring. Examples of
CA 02512183 2005-06-29
19
-(Cg-C~4)-aryl radicals are phenyl, naphthyl, for example 1-naphthyl and
2-naphthyl, biphenylyl, for example 2-biphenylyl, 3-biphenylyl and
4-biphenylyl, anthryl or fluorenyl. Biphenylyl radicals, naphthyl radicals
and,
in particular, phenyl radicals are preferred aryl radicals.
The term "Het" is understood as meaning a saturated or unsaturated
monocyclic or bicyclic, 3- to 10-membered heterocyclic ring system which
contains 1, 2 or 3 identical or different ring heteroatoms from the series
nitrogen, oxygen and sulfur. In the underlying monocyclic or bicyclic
heterocyclic ring system, Het contains 3, 4, 5, 6, 7, 8, 9 or 10 ring atoms.
The monocyclic ring system can be a 3-, 4-, 5-, 6- or 7-membered ring. In
the bicyclic Het, two rings can be linked to each other, with it being
possible
for one of the rings to be a 5-membered or 6-membered heterocyclic ring
and the other to be a 5- or 6-membered heterocyclic or carbocyclic ring. A
bicyclic Het group can, for example, be composed of 8, 9 or 10 ring atoms.
Het comprises saturated heterocyclic ring systems which do not possess
any double bond in the rings and also unsaturated heterocyclic ring
systems, including monounsaturated and polyunsaturated heterocyclic ring
systems, which possess one or more double bonds and form a stable ring
system. Unsaturated rings can be partially unsaturated or form an aromatic
system. The Het group contains identical or different heteroatoms from the
series nitrogen, oxygen and sulfur. Examples of heterocycles from which
the Het group can be derived are the radicals acridinyl, benzimidazolyl,
benzofuranyl, benzothiofuranyl, benzothiophenyl, benzoxazolyl,
benzothiazolyl, benzotriazolyl, benzotetrazolyl, benzisoxazolyl,
benzisothiazolyl, benzimidazalinyl, carbazolyl, 4aH-carbazolyl, carbolinyl,
chromanyl, chromenyl, cinnolinyl, decahydroquinolinyl, 2H, 6H-1,5,2-dithia-
zinyl, dihydrofuran[2,3-b]tetrahydrofuran, fuaranyl, furazanyl,
imidazolidinyl,
imidazolinyl, imidazolyl, 1 H-indazolyl, indolinyl, indolizinyl, indolyl, 3H-
indolyl, isobenzofuranyl, isochromanyl, isoindazolyl, isoindolinyl,
isoindolyl,
isoquinolinyl (benzimidazolyl), isothiazolyl, isoxazolyl, morpholinyl,
naphthyridinyl, octahydroisoquinolinyl, oxadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-
oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, oxazolidinyl, oxazolyl,
oxazolidinyl, pyrimidinyl, phenanthridinyl, phenanthrolinyl, phenazinyl,
phenothiazinyl, phenoxathiinyl, phenoxazinyl, phthalazinyl, piperazinyl,
piperidinyl, pteridinyl, purynyl, pyranyl, pyrazinyl, pyroazolidinyl,
pyrazolinyl,
pyrazolyl, pyridazinyl, pyridooxazoles, pyridoimidazoles, pyridothiazoles,
pyridinyl, pyridyl, pyrimidinyl, pyrrolidinyl, pyrrolinyl, 2H-pyrrolyl,
pyrrolyl,
quinazolinyl, quinolinyl, 4H-quinolizinyl, quinoxalinyl, quinuclidinyl,
tetrahydrofuranyl, tetrahydroisoquinolinyl, tetrahydroquinolinyl, 6H-1,2,5-
CA 02512183 2005-06-29
thiadazinyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4-
thiadiazolyl, thianthrenyl, thiazolyl, thienyl, thienothiazolyl,
thienooxazolyl,
thienoimidazolyl, thiophenyl, triazinyl, 1,2,3-triazolyl, 1,2,3-tirazolyl,
1,2,4-
triazolyl, 1,2,5-triazolyl, 1,3,4-tirazolyl and xanthenyl.
5 Preference is given to pyridyl; such as 2-pyridyl, 3-pyridyl or 4-pyridyl;
pyrrolyl; such as 2-pyrrolyl and 3-pyrrolyl; furyl; such as 2-furyl and 3-
furyl;
thienyl; such as 2-thienyl and 3-thienyl; imidazolyl; pyrazolyl, oxazolyl,
isoxazolyl, thiazolyl, isothiazolyl, tetrazolyl, pyridazinyl, pyrazinyl,
pyrimidinyl, indolyl, isoindolyl, benzofuranyl, benzothiophenyl, 1,3-benzo-
10 dioxolyl, indazolyl, benzimidazolyl, benzoxazolyl, benzothiazolyl,
quinolinyl,
isoquinolinyl, chromanyl, isochromanyl, cinnolinyl, quinazolinyl,
quinoxalinyl, phthalazinyl, pyridoimidazolyl, pyridopyridinyl,
pyridopyrimidinyl, purinyl and pteridinyl.
15 Particular preference is given to a Het from the group aziridine, oxirane,
azetidine, pyrrole, furan, thiophene, dioxole, imidazole, pyrazole, oxazole,
isoxazole, thiazole, isothiazole, 1,2,3-triazole, 1,2,4-triazole, pyridine,
pyran, thiopyran, pyridazine, pyrimidine, pyrazine, 1,4-dioxin, 1,2-oxazine,
1,3-oxazine, 1,4-oxazine, 1,2-thiazine, 1,3-thiazine, 1,4-thiazine,
20 1,2,3-triazine, 1,2,4-triazine, 1,3,5-triazine, azepine, 1,2-diazepine,
1,3-diazepine, 1,4-diazepine, indole, isoindole, benzofuran, benzothio-
phene, 1,3-benzodioxole, benzo[1,4]dioxin, 4H-benzo[1,4]oxazine,
indazole, benzimidazole, benzoxazole, benzothiazole, quinoline,
isoquinoline, chroman, isochroman, cinnoline, quinazoline, quinoxaline,
phthalazine, pyridoimidazoles, pyridopyridines or pyridopyrimidines, etc.
and also ring systems which ensue from the listed heterocycles by linking
to or annelation with a carbocyclic ring, for example benzoanelated,
cycopentaanelated, cycohexaanelated or cycloheptaanelated derivatives of
these heterocycles. Suitable nitrogen heterocycles can also be present as
N oxides or as quaternary salts in which a suitable nitrogen atom is
alkylated with -(C~-C4)-alkyl radicals. The Het groups can be unsubstituted
or substituted in accordance with the listed definitions.
The term "osteoarthrosis" is understood as meaning a disease which
chiefly develops in connection with a disparity between the strain on and
the load capacity of the individual joint components and joint tissues, which
is associated with increasing destruction of the cartilage and which is in the
main not inflammatory. Damage to the joint cartilage, such as fraying,
demedullation and hyalinization, followed by reactive changes in the
CA 02512183 2005-06-29
21
subchondral bone, and also capsule changes, is prominent in the
pathology. The term "spondylosis" is understood as meaning an arthrosis of
the vertebral bodies, with this arthrosis being characterized by a
noninflammatory loss of cartilage from the vertebral bodies and
intervertebral disks.
The compounds of the formula I can be prepared, for example, by reacting
a compound of the formula II
0
-Y
N
III)
N C-Y
O
a) with a compound of the formula Illa or Illb
R1
Illa R5 ~ ~ NHz Illb
R2
R6 __
where R1, R2, R3, R4, R5, R6 and R7 have the meanings given in
formula I and Y is halogen, hydroxyl or C~-C4-alkoxy or forms,
together with the carbonyl group, an active ester or a mixed
anhydride, with a compound of the formula I being formed and the
reaction products being converted, where appropriate, into their
physiologically tolerated salts, or
b) reacting a compound of the formula II with a compound of the
formula Illa or Illb to give a compound of the formula IVa or IVb
CA 02512183 2005-06-29
22
0 0
0 0
\ Y
N
R1 ~N ( \ Y H NON
NON
R2
(IVa) (IVb)
where R1 to R7 have the meanings given in formula I and Y is
halogen, hydroxyl or C~-C4-alkoxy or forms, together with the
carbonyl group, an active ester or a mixed anhydride, and purifying
the compound of the formula IVa or IVb, where appropriate, and
then using a compound of the formula Illa or Illb to convert it into a
compound of the formula I.
The preparation of compounds of the formula I, and the preparation of the
starting substances which are required for this purpose - insofar as these
substances are not commercially available - are described in more detail
below.
The compounds according to the invention are most easily prepared by
mixing the two components, i.e. the pyrimidine derivative of the formula (II)
and the amine of the formula Ills or Illb in equimolar quantities and reacting
them, at temperatures of from -30°C to 150°C, preferably at from
20°C to
100°C, to give a compound of the formula IVa or IVb and then reacting
the
compounds of the formula IVa or IVb with up to an equimolar quantity of
the amine of the formula Illb or Illa in an analogous manner. The
termination of the reaction can be determined, for example, by means of
thin layer chromatography or HPLC-MS. A variant of this method is that the
reaction is carried out in a suitable solvent, such as diethyl ether,
dimethoxyethane or tetrahydrofuran, chlorinated hydrocarbons, such as
methylene chloride, choloroform, trichloroethylene or tetrachloroethylene,
benzene or toluene, or else polar solvents, such as dimethylformamide,
acetone or dimethyl sulfoxide. The reaction temperatures in this connection
are between room temperature and the boiling point of the solvent, with
temperatures in the range from room temperature up to 130°C being
particularly preferred.
Reaction can also take place by way of a mixed anhydride, such as ethyl
chloroformate, or by way of an active ester, such as paranitrophenyl ester
CA 02512183 2005-06-29
23
(Y = CICH2-COO or N02-CgH4-O). Appropriate methods are known and
described in the literature.
A compound of the formula II or a compound of the formula IVa or IVb can
also react with an amine of the formula Illa or Illb if Y is OH and the
corresponding carboxylic acid is activated in situ using customary coupling
reagents. Examples of these coupling reagents are carbodiimides, such as
dicyclohexylcarbodiimide (DCC) or diisopropylcarbodiimide (DCI), or
N,N'-carbonyldiazoles, such as N,N'-carbonyldiimidazole, or a uronium salt,
such as O-((cyano(ethoxycarbonyl)methylene)amino)-1,1,3,3-tetramethyl-
uroniumtetrafluoroborate (TOTU) or O-(7-azabenzotriazol-1-yl)-
1,3,3,-tetramethyluronium hexafluorophosphate (HATU). Appropriate
methods are known.
If amines of the formula Illa or Illb are not commercially available, they can
be prepared from corresponding, commercially available starting
compounds using methods known in the literature. Examples of suitable
starting compounds for amines are nitrites, nitro compounds,
carboxamides, carboxylic esters, carboxylic acids, aldehydes and
bromides. Nitrites, nitro compounds and carboxamides can be reduced to
amines using known methods. Carboxylic acids and carboxylic esters can
be converted into the carboxamides. Aldehydes can be converted directly
into the amines by means of a reductive amination with NH4Ac/NaBH4, or
can be initially converted into the oximes using hydroxylamine and then
being converted into the amines by reduction.
Where appropriate, the reaction can also take place in the presence of
bases. Examples of suitable additional bases are carbonates or hydrogen
carbonates, such as sodium carbonate or potassium carbonate or sodium
hydrogen carbonate or potassium hydrogen carbonate, or tertiary amines,
such as triethylamine, tributylamine or ethyldiisopropylamine, or
heterocyclic amines, such as N-alkylmorpholine, pyridine, quinoline or
dialkylanilines.
Where appropriate, the products, in particular the compound of the formula
IVa or IVb can be worked up, for example, by means of extraction or
chromatography, e.g. through silica gel. The isolated product can be
recrystallized and reacted, where appropriate, with a suitable acid to give a
physiologically tolerated salt. Examples of suitable acids are:
CA 02512183 2005-06-29
24
mineral acids, such as hydrochloric acid and hydrobromic acid, and also
sulfuric acid, phosphoric acid, nitric acid or perchloric acid, or organic
acids,
such as formic acid, acetic acid, propionic acid, succinic acid, glycolic
acid,
lactic acid, malic acid, tartaric acid, citric acid, malefic acid, fumaric
acid,
phenylacetic acid, benzoic acid, methanesulfonic acid, toluenesulfonic acid,
oxalic acid, 4-aminobenzoic acid, naphthalene-1,5-disulfonic acid or
ascorbic acid.
Insofar as they are not commercially available, the starting compounds of
the formula Illa or Illb can be prepared readily (e.g. Organikum, Organisch
Chemisches Grundpraktikum [Organicum, basic practical course in organic
chemistryJ, 15th edn, VEB Deutscher Verlag der Wissenschaften, 1976; the
methods index contains a review of the different possibilities on p. 822).
The starting compounds of the formula (II) are obtained, for example, by
reacting pyrimidine-4,6-dicarboxylic acid, or pyridine-2,4-dicarboxylic acid,
to give the corresponding pyrimidine-4,6-dicarbonyl halide or pyridine-2,4-
dicarbonyl halide, preferably chloride (using methods known from the
literature), preferably in the presence of a catalyst such as dimethyl-
formamide. This acid halide can then be reacted, for example, either with a
suitable alcohol, e.g. paranitrobenzyl alcohol to give the corresponding
active ester, or else with lower alcohols, such as methanol or ethanol, to
give the corresponding esters. The pyrimidine-4,6-dicarboxylic acid can
also initially be converted, in the added presence of a suitable carboxylic
acid or of a carboxylic ester, such as ethyl chloroformate, into a mixed
anhydride, which is then reacted with the amines of the compound of the
formulae Illa or Illb and IVa or IVb to give the products according to the
invention. A corresponding method is also described in the literature.
The pyrimidine-4,6-dicarboxylic acid is prepared in accordance with
methods known from the literature, for example by oxidizing 4,6-dimethyl-
pyrimidine, which, for its part, can be obtained, for example, by
catalytically
hydrogenating commercially available 2-mercapto-4,6-dimethylpyrimidine.
Insofar as compounds of the formula I permit diastereoisomeric or
enantiomeric forms and accrue as their mixtures in the synthesis which is
selected, separation into the pure stereoisomers is achieved either by
chromatography on an optionally chiral support material or, provided the
racemic compound of the formula I is capable of salt formation, by
CA 02512183 2005-06-29
fractionally crystallizing the diastereomeric salts which are formed using an
optically active base or acid as auxiliary substance. Examples of suitable
chiral stationary phases for the thin-layer or column chromatographic
separation of enantiomers are modified silica gel supports (what are termed
5 Pirkle phases) and also high molecular weight carbohydrates, such as
triacetyl cellulose. Following appropriate derivatization, which is known to
the skilled person, gas-chromatographic methods on chiral stationary
phases can also be used for analytical purposes. In order to separate the
enantiomers of the racemic carboxylic acids, the diastereomeric salts,
10 which differ in solubility, are formed using an optically active, as a rule
commercially available, base such as (-)-nicotine, (+)- and (-)-
phenylethylamine, quinine bases, L-lysine or L- and D-arginine, the more
sparingly soluble component is isolated as a solid, the more readily soluble
diastereomer is separated out from the mother liquor, and the pure
15 enantiomers are isolated from the diastereomer salts which have been
obtained in this way. The racemic compounds of the formula I which
contain a basic group, such as amino group, can, in what is in principle the
same manner, be converted into the pure enantiomers using optically
active acids, such as (+)-camphor-10-sulfonic acid, D- and L-tartaric acid,
20 D- and L-lactic acid and (+) and (-)-mandelic acid. Chiral compounds which
contain alcohol or amine functions can also be converted into the
corresponding esters or amides using appropriately activated or optionally
N-protected enantiomerically pure amino acids or, conversely, chiral
carboxylic acids can be converted into the amides using carboxyl-protected
25 enantiomerically pure amino acids or into the corresponding chiral esters
using enantiomerically pure hydroxycarboxylic acids such as lactic acid.
The chirality of the amino acid or alcohol radical which has been introduced
in enantiomerically pure form can then be used for separating the isomers
by the diastereomers, which are now present, being separated by means of
crystallization or chromatography on suitable stationary phases and, after
that, using suitable methods to once again eliminate the entrained chiral
molecule moiety.
Acidic or basic products of the compound of the formula I may be present
in the form of their salts or in free form. Preference is given to
pharmacologically tolerated salts, e.g. alkali metal salts or alkaline earth
metal salts or hydrochorides, hydrobromides, sulfates, hemisulfates, all
possible phosphates and also salts of the amino acids, natural bases or
carboxylic acids.
CA 02512183 2005-06-29
26
Physiologically tolerated salts are prepared in a manner known per se from
compounds of the formula I, including their stereoisomeric forms, which are
capable of salt formation. The carboxylic acids form stable alkali metal
salts, alkaline earth metal salts or optionally substituted ammonium salts
with basic reagents such as hydroxides; carbonates, hydrogencarbonates,
alkoxides and ammonia or organic bases, for example trimethylamine,
triethylamine, ethanolamine or triethanolamine, or else basic amino acids,
for example lysine, ornithine or arginine. Insofar as the compounds of the
formula I possess basic groups, stable acid addition salts can also be
prepared using strong acids. Both inorganic and organic acids, such as
hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid,
methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid,
4-bromobenzenesulfonic acid, cyclohexylamidosulfonic acid,
trifluoromethylsulfonic acid, acetic acid, oxalic acid, tartaric acid,
succinic
acid and trifluoroacetic acid are suitable for this purpose.
As a result of their pharmacological properties, the compounds of the
formula I are suitable for the prophylaxis and therapy of all those diseases
whose course involves an increased activity of matrix metalloproteinase 13.
These diseases include degenerative joint diseases, such as
osteoarthroses, spondyloses and cartilage loss following joint trauma or
relatively long joint immobilization following meniscus or patella injuries or
ligament rupture. They also include diseases of the connective tissue, such
as collagenoses, periodontal diseases, wound healing disturbances and
chronic diseases of the locomotory apparatus such as inflammatory,
immunologically determined or metabolism-determined, acute and chronic
arthritides, arthropathies, myalgias and disturbances of bone metabolism,
or cancers, such as breast cancer.
The pharmaceuticals according to the invention can be administered by
subcutaneous, intraarticular, intraperitoneal or intravenous injection.
Intraarticular injection is preferred. Rectal, oral, inhalative or transdermal
administration is also possible.
The invention also relates to a process for producing a pharmaceutical,
wherein at least one compound of the formula I is brought, together with a
pharmaceutically suitable and physiologically tolerated carrier and, where
CA 02512183 2005-06-29
27
appropriate, other suitable active compounds, additives or auxiliary
substances, into a suitable form for administration.
The compounds of the formula I are mixed with the additives, such as
carrier substances, stabilizers or inert diluents, which are suitable for the
purpose and brought into suitable administration forms, such as tablets,
sugar-coated tablets, hard gelatin capsules, aqueous, alcoholic or oily
suspensions or aqueous or oily solutions, using the customary methods.
Examples of inert carrier substances which can be used are gum arabic,
magnesium oxide, magnesium carbonate, potassium phosphate, lactose,
glucose or starch, in particular corn starch. In this connection, the
preparation can be effected either as dry granules or as wet granules.
Examples of suitable oily carrier substances or solvents are vegetable or
animal oils, such as sunflower oil or cod liver oil.
For subcutaneous, intraarticular, intraperitoneal or intravenous
administration, the active compounds are, if desired, brought into solution,
suspension or emulsion using the substances which are suitable for this
purpose, such as solubilizers, emulsifiers or other auxiliary substances.
Examples of suitable solvents are physiological sodium chloride solution or
alcohols, e.g. ethanol, propanol or glycerol, and, in addition, sugar
solutions, such as solutions of glucose or mannitol, or else a mixture which
is composed of the different solvents mentioned.
Customary adjuvants, such as carrier substances, disintegrants, binding
agents, coating agents, swelling agents, glidants or lubricants, flavorings,
sweeteners and solubilizers are also used. Auxiliary substances which are
frequently employed and 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, peanut oil or sesame seed oil,
polyethylene glycol and solvents such as sterile water and monohydric or
polyhydric alcohols, such as glycerol.
The compounds of the formula I are preferably prepared as pharmaceutical
preparations and administered in dosage units, with each unit containing a
defined dose of the compound of the formula I as the active constituent.
For this purpose, the compounds of the formula I can be administered
orally in doses of from 0.01 mg/kg/day to 25.0 mg/kg/day, preferably from
CA 02512183 2005-06-29
28
0.01 mg/kg/day to 5.0 mg/kg/day, or parenterally in doses of from
0.001 mg/kg/day to 5 mg/kg/day, preferably of from 0.001 mg/kg/day to
2.5 mg/kg/day. The dosage can also be increased in severe cases.
However, smaller doses are also adequate in many cases. These figures
relate to the treatment of an adult.
The invention is explained in more detail below with the aid of examples.
Example 1: tert-Butyl [4-(f[6-(4-fluoro-3-methylbenzylcarbamoyl)pyrimidine-
4-carbonyl]amino}methyl)phenoxy]acetate
tert-Butyl (4-formylphenoxy)acetate: 10 g (0.0819 mol) of 4-hydroxybenz-
aldehyde and 15.97 g (0.0819 mol) of tert-butyl bromoacetate were
dissolved in 200 ml of 2-butanone, after which 11.32 g (0.0819 mol) of
potassium carbonate were added and the mixture was heated under reflux
for 2 hours (h). The mixture was then concentrated under reduced pressure
and the residue was taken up in water and the solution was extracted three
times with dichloromethane. The organic phase was dried (MgS04), filtered
and concentrated under reduced pressure.
Yield: 18.72 g (97%)
tert-Butyl [4-(hydroxyiminomethyl)phenoxy]acetate: 18.72 g (0.0792 mol) of
tert-butyl (4-formylphenoxy)acetate were dissolved in 200 ml of
water/ethanol (1:1), after which 6.056 g (0.0872 mol) of hydroxyammonium
chloride and 3.169 (0.0792 mol) of sodium hydroxide were added and the
mixture was stirred under reflux for 2.5 hours (h). The mixture was
concentrated under reduced pressure and the residue was taken up in
water and the solution was extracted with ethyl acetate. The organic phase
was dried (MgS04), filtered and concentrated under reduced pressure.
Yield: 19.9 g (100%) MS (ES+): m/e = 251.12
tert-Butyl (4-aminomethylphenoxy)acetate: 6 g (0.0239 mol) of tert-butyl [4-
(hydroxyiminomethyl)phenoxy]acetate were dissolved in 10 ml of
water/ethanol (1:1 ) and hydrogenated, under a pressure of 5 bar and at
room temperature, over 0.06 g of Pd/C (10%). After 3 hours (h), the mixture
was filtered through Celite and concentrated under reduced pressure.
Yield: 4.26 g (75%)
CA 02512183 2005-06-29
29
Methyl 6-(4-fluoro-3-methylbenzylcarbamoyl)pyrimidine-4-carboxylate: the
compound was prepared in analogy with methyl 6-benzylcarbamoylpyrimi-
dine-4-carboxylate.
6-(4-Fluoro-3-methylbenzylcarbamoyl)pyrimidine-4-carboxylic acid: 0.889 g
(0.0222 mol) of sodium hydroxide in 6 ml of water was added to 8.75 g
(0.0202 mol) of methyl 6-(4-fluoro-3-methylbenzylcarbamoyl)pyrimidine-4-
carboxylate in 80 ml of methanol and the mixture was stirred at room
temperature (RT). After 2.5 hours, the solvent was removed under reduced
pressure and the residue was dissolved in 300 ml of water and the solution
was acidified with concentrated HCI. The precipitate was filtered off and
dried.
Yield: 5.43 g (95%) MS (ES+): m/e = 289.09
tert-Butyl [4-({[6-(4-fluoro-3-methylbenzylcarbamoyl)pyrimidine-4-carbonyl]-
amino}methyl)phenoxy]acetate: 150 mg (0.519 mmol) of 6-(4-fluoro-3-
methylbenzylcarbamoyl)pyrimidine-4-carboxylic acid were dissolved in 5 ml
of DMF, after which 170 mg (0.519 mmol) of TOTU were added and the
mixture was stirred at RT for 30 minutes. 147.6 mg (0.62 mmol) of tert-butyl
(4-aminomethylphenoxy)acetate and 119.55 mg (1.038 mmol) of N-ethyl-
morpholine were then added and the mixture was stirred at RT for 12
hours. The solvent was removed under reduced pressure and the residue
was purified by preparative HPLC (water/acetonitrile). The homogeneous
fractions were concentrated under reduced pressure and freeze-dried.
Yield: 126 mg (48%) MS (ES-): m/e = 508.21
Example 2: [4-({[6-(4-Fluoro-3-methylbenzylcarbamoyl)pyrimidine-4-carbonyl]-
amino}methyl)phenoxy]acetic acid
83.6 mg (0.16 mmol) of tert-butyl [4-({[6-(4-fluoro-3-methylbenzylcarba
moyl)pyrimidine-4-carbonyl]amino~methyl)phenoxy]acetate (Example 31 )
were stirred at RT for 4 hours in 90% trifluoroacetic acid. Acetonitrile/water
was then added and the precipitate was filtered off and dried.
Yield: 55 mg (76%) MS (ES+): m/e = 452.15
The following compounds were prepared in analogy with Example 1.
CA 02512183 2005-06-29
30
Table 1:
Example Structure MS (ESI+)
1 508.21
0
o' v /
I
\ N/~ N / I F
NI / H \
0 0
2 452.15
w
o~
o \
I ~ N~ N
I
O O
3 477.00
N~ f
N/~N / F
N I / N \ I
O O
4 ~0 396.13
S N~N /
N I / N \ I ~~a
O
O O
5 H,c 463.OZ
s
N~N / F
N~~N
0 0
6 382.11
s
o \ /N o-
N
O
7 ~ 384.11
i'~NI / I F
N~~N
I0 0O
8 N/ N~N / F 433.05
a \ I N I / N \ ( a
0 0
9 411.11
N I NI~N / I
CI ~ N N ~ O~CH~
O O
CA 02512183 2005-06-29
31
10 ~c~ 407.16
N~N
\ I N I ~ N \
O O
11 H,c~o 473.05
N ~ I Br
\ N~ N / F I
N I / N \ I
O O
1 Z S 6r 448.00
N~N ~ F
N I~~N ~
O O
13 380.15
~~ lol i N ~ I ~
o~c
0 0
14 ~ ~ F - 399.09
~ I N I ~ N
G
0 0
15 ~ , F 404.05
I N I / N ~
a
0 0
16 382.14
Gi,
O N/~N / F
N I / N
O O
17
443.04
N 9r
\ I N I/~N N / I F
/ \
O O
18 448.00
/i
i~N / I F
N / N \
O O
CA 02512183 2005-06-29
32
19 ~O 418.07
N~ / F
N I / N \
CI
O O
2p o~~ 429.10
F
N/ N ~N /
\ I N ( / N \ d
O O
21 382.11
NON /
N I / \
0 0
22 377.15
N/ I ~~N ~ I
\ N / N \ ~Cfi3
O
O O
23 ~ F 404.05
N N /
N 4 / N \ I a
0 0
24 a N 477.00
~f
/ N/~N / F
N I / N \
O O
25 ~,c 449.03
sr
NON / F
N I / N ~
0 O
26 ~ I cri, 368.13
F
~hN / I
N"~~N \
'O~ ~ ~O
27 F 443.04
% N~N
\ ~ N I / N'Wgr
O O
CA 02512183 2005-06-29
33
28 F 418.07
N~N /
\ N ~ / N \ I d
O 0
Z9 ~, 473.05
O N
I ~ r
NON / F
N
O O
30 377.15
N N/~N /
\ I N I / N~O
1
O O
31 _ 366.13
I O I N I /N N \ ( i~
O
O O
32 "-- H 382.16
HOC-N
N/~N ~ F
N ~ / N
0 0
33 ~ - ~ 409.15
O i N~N / F
\ I N I / N \ I
O O
446.05
N
Br
i~N / I F
N / N \
O O
35 - 398.1 Z
I w NON S
i N I ~ N
0 O O
CA 02512183 2005-06-29
34
36 396.00
NII~N
N~~N \ ~ Oil
O~ ~ ~O
37 ~ F 418.07
s N ~N /
/ N I / N
G
C
f'Ls 0 0
38 396.13
N~N /
~~N ( / N \ I OiC~
hi~CIY \/ \
39 / ~ 379.14
N~ N~N / F
N I / N \ I
0 0
40 ~i B~ 449.00
</ F
S~ N~N /
N I / N \ I
O O
41 H,c N~ cH 394.16
I
~N N~N / F
N I / N \
O O
42 F 394.08
N~ N /
i0 N I / N \
H,C ~ ~~~ G
0 0 0
43 405.05
F
N N~N
~N I / N \ I G
O 0
44 383.00
N N~N /
~~N I / N \ I O~C~
O O
CA 02512183 2005-06-29
35
45 F / N/yN N 365.13
\ I N I / N \
0 O
46 Ha 393.16
F
N~N /
wN ~ N~~~N \ I
O~ '' ~O
47 ~H~ 388.15
H C O\ ~ / F
~ ~O IN NI /N N \ I
O O
48 380.07
~N / F
HO N I / N \ I
O O O
49 392.16
i
~C~N~N I /N \
O O
50 399.09
N~N / F
N ~ N ~ / N \ d
0 O
51 434.16
O N~N
~CwO~N N \
Clip O 0
52 391.16
N~N /
N \ N I / N \ I OiC~
( /
CA 02512183 2005-06-29
36
5~- ' 358.13
0 N~N
~C~O~N I / N \
O O
54 F 380.07
I1~C N / N
I~N / ~ G
O O
55 ~c~ 360.12
cH,
I~ N / I F
N~~N \
O O
56 394.08
F
O N~N /
~C~O N I / N \ ~ d
Giy O O
57 392.16
\ ~ N I / N
N~ N~N
N
O O
5g 391.16
N~N
N~~N \
O O
5g 408.15
N~N
~N. N~~~N \ I OiC
[/
N O O
hi~C
60 372.14
N~N /
N I / N
0 O O
CA 02512183 2005-06-29
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61 ~o\ 346.14
N~ / F
N~~N \ I
O O
62 393.00
'1
\ N
N~N / F
N~~N \ I
O 0
63 ~~, 461.00
0
N
\ /
NInN / I F
N'~~N \
O O
6q OH CH3 332.13
N N / F
N I / N \
O O
65 N--, 382.16
N
N~N / F
N~~N \
O 0
66 330.13
N~N
HON I / N \ I O~C~
O 0
CA 02512183 2005-06-29
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67 N, 434.03
Br
vN N~N / F
N I / N \
O O
68 H,~~N~~, S19.12
O
Br
~C~O~ N~N ~ ~ F
N / N
O O
69 ~ 463.22
w I r~ N ~ w
0
0 0
70 \~ 463.00
N~ N /
\ I " i ~- b \ I
0
0 0
71 477.24
I
/N
N 'N /
\ I / \ ~ O/
O O
72 527.17
\ ~' N \ i F
a
0 0
CA 02512183 2005-06-29
39
73 420.14
ai
O ~N
\( ~ \I
0
0 0
74 452.15
o\ / a-i
I
~N / I F
/ \
O O
75 505.23
N~ N
~i y ~I
J o
0 0
76 a 416.07
/
N-~'~
\ /N
O
O
O
77 465.05
s e~
i ' N / I F
t-N / N \
O O
78 399.22
s
~N / F
~f ' \
O O
CA 02512183 2005-06-29
40
79 413.11
a
N~
\ N~ N / I F
I / H \
0 0
80 418.07
F
I ~ N~ N
O O
81 / 380.15
N
O
NH _
O
O-
82 399.09
N~ N / F
\ I H I / \ a
O O
83
404.05
~ F
I I N i N H
a
0 0
84
382.14
N~ N / F
i /
0 0
85 443.04
N~
I
\ ~ ~ F
~-r~ I , b \ I
0 0
CA 02512183 2005-06-29
41
86 448.00
j S 8'
/ i 'N / I F
/ \
O O
87 418.07
5 H i 'N H / I F
/ N~.,
a
0 0
gg 384.11
r~~ N /
/ ~ \
O O
89 377.15
N~ N / I
H I N \ O/
0 O
90 404.05
N~ N / F
I / \
a
0 0
91 477.00
a
I
F
I / H \
O O
92 379.14
F
N \\~N
I / N ~ / N ~~~I .
O O
93 449.03
o a
/ F
I/
I
CA 02512183 2005-06-29
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Pharmacological examples
Determining the enzyme activity of the catalytic domain of human
collagenase 3 (MMP-13).
This protein is obtained as an inactive proenzyme from INVITEK, Berlin
(Catalog No. 30 100 803). Activating the proenzyme:
2 parts by volume of proenzyme are incubated with 1 part by volume of
APMA solution at 37°C for 1.5 hours. The APMA solution is prepared
from
a 10 mmol/l solution of p-aminophenylmercuric acetate in 0.1 mmol/l NaOH
by diluting with 3 parts by volume of tris/HCl buffer, pH 7.5 (see below).
The pH is adjusted to between 7.0 and 7.5 by adding 1 mmol/I HCI. After
the enzyme has been activated, it is diluted to a concentration of
1.67 ~g/ml using the tris/HCl buffer.
In order to measure the enzyme activity, 10 NI of enzyme solution are
incubated for 15 minutes with 10 NI of a 3% (v/v) buffered solution of
dimethyl sulfoxide (reaction 1). In order to measure the enzyme inhibitor
activity, 10 NI of enzyme solution are incubated with 10 NI of a 3% (v/v)
buffered solution of dimethyl sulfoxide containing the enzyme inhibitor
(reaction 2).
In the case of both reaction 1 and reaction 2, the enzyme reaction is
monitored by fluorescence spectroscopy (328 nm (extinction)1393 nm
(emission)) after adding 10 NI of a 3% (v/v) aqueous solution of dimethyl
sulfoxide containing 0.75 mmol of the substrate/I.
The enzyme activity is depicted as increase in extinction/minute.
The effect of the inhibitor is calculated as a percentage inhibition using the
following formula:
inhibition = 100 - [(increase in extinction/minute in reaction 2)/(increase
in extinction/minute in reaction 1 ) x 100].
The ICSp, i.e. the inhibitor concentration which is required for inhibiting
the
enzyme activity by 50%, is determined graphically by plotting the
percentage inhibitions at different inhibitor concentrations.
The buffer solution contains 0.05% Brij (Sigma, Deisenhofen, Germany)
and
0.1 mol of tris/HCl/l, 0.1 mol of NaCl/l and 0.01 mol of CaCl2/I (pH = 7.5).
The enzyme solution contains 1.67 ~,g of the enzyme domain/ml.
CA 02512183 2005-06-29
43
The substrate solution contains 0.75 mmol/l of the fluorogenic substrate (7-
methoxycoumarin-4-yl)acetyl-Pro-Leu-Gly-Leu-3-(2',4'-dinitrophenyl)-L-2,3-
diaminopropionyl-Ala-Arg-NH2 (Bachem, Heidelberg, Germany).
Table 2 below shows the results.
Table 2:
Example IC5p MMP13 (nM) Example IC5p MMP13 (nM)
1 12 20 50
2 7 28 100
4 24 35 200
11 30 44 500
Determining the enzyme activity of the catalytic domain of human
neutrophil collagenase (MMP-8) and of human stromelysin (MMP-3).
The enzymes human neutrophil collagenase and human Stromelysin were
prepared as active catalytic domains as described in Weithmann et al
Inflamm Res, 46 (1997), pages 246-252. The measurement of the enzyme
activity, and the determination of the inhibitory effect of inhibitors on the
enzyme activity, were also carried out as described in that publication.
When determining human neutrophil collagenase and human stromelysin,
the compounds described in the above examples in each ~ case had IC50
values of more than 100 000 nM. These compounds are therefore virtually
without activity as regards inhibiting MMP 3 and 8.
Determining prolyl hydroxylase inhibition by the method of Majamaa Eur. J.
Biochem. 138 (1984) 239-245 using the version of Kaule and Guenzler
Analytical Biochemistry 184 (1990) 291-297.
0.03 pg of prolyl 4-hydroxylase, which was prepared, by the method of
Kedersa, Collagen Relat. Res. 1 (1981 ) 345-353, from homogenized 14-
day-old chick embryos by ammonium sulfate precipitation and affinity
chromatography on poly-L-proline-coupled Sepharose 4B and subsequent
DEAE cellulose chromatography, was incubated, at 37°C for one
hour, in
0.05 ml of an 0.04 M tris-HCI solution (pH = 7.5) containing 0.05 mM
CA 02512183 2005-06-29
44
FeS04, 0.1 mM 2-oxo-[5-C-14]glutarate (100 000 dpm), 20 pg (0.4 mg/ml)
of (Pro-Pro-Gly)~p (Protein Research Foundation, Minoh, Osaka, Japan),
1 mM ascorbic acid, 0.4 mg of catalase/ml, 0.5 mM dithiothreitol and 2 mg
of bovine serum albumin/ml, as well as the concentration of the inhibitor to
be measured (the comparison experiment was carried out without any
inhibitor). After that, 25 pl of a 20 mM succinate and 2-oxaglutarate
solution, and also 25 pl of a 0.16 M 2,4-DNPH solution containing 30%
HC104, were added. After the mixture had been incubated at room
temperature for a further 30 minutes, it was centrifuged at 3000 g for 5
minutes. The radioactivity of 100 p.l of the supernatant was determined by
liquid scintillation. Production of the [1-C-14] succinate was given in dpm.
Unless otherwise indicated, all the chemicals and reagents were obtained
from Sigma, Sigma D-82024 Taufkirchen.
The compounds tested in Table 3 were used at concentrations of from
0.1 wM to 1 mM. Where it was possible, the IC50 value was determined by
graphic extrapolarization of the individual results. This value denotes the
concentration of active inhibitors which led to 50% inhibition of the enzyme.
Table 3:
Example Prolyl hydroxylaseExample Prolyl hydroxylase
IC5 M ICS M
Pyrimidine-4,6-1.2 70 No inhibition
dicarbox lic
acid
7 No inhibition 71 No inhibition
14 No inhibition 72 No inhibition
16 No inhibition 83 No inhibition
22 No inhibition 84 No inhibition
69 No inhibition
