Note: Descriptions are shown in the official language in which they were submitted.
2 ~3~ t~
HOECHST ~XTIENGESELLSCHAFT HOE 90~F 295 Dr. FI/rh
Description
4- or 5-sub~tituted pyridine-2-carboxylic acids, a
proce~s for the preparstion thereof and the U8Q thereof
a~ pharmaceuticals
Compounds which inhibit the enzymes proline hydroxylase
and lysine hydroxyla~e effect very æelecti~e inhibition
sf collagen biosynthesis due to influencin~ ~ollagen-
specific hydroxylation reactions. In the course thereof,
protein-bound proline or ly~ine i-s hydro*ylated b~ the
enzymes proline hydroxyla~e or lysine hydro~ylase respec-
tively. If this reaction is suppressed by inhibitors, the
resulting collagen molecule is unable to function, is
insufficiently hydroxylated and can be released by the
cells only in small amounts into the extracellular spase.
The insufficiently hydroxylated collagen i5 unable,
moreover, to be incorporated ln ~he collagen matrix and
is ve~y easily broken down by proteolysis. The conse-
~uence of these effects is an overall reduction in the
amount o~ collagen deposited in the extracellular spase.
It is known that inhibition of proline hydr~xyla~e ~y
known inhihitor~ ~uch as ~ dipyridyl results în
inhibition of Clq biosynthesis by macrophages (W. Muller
et al., FEBS Lett. 90 (1978), 218; Immunobiolo~y 155
2S (1978), 47~. This results in the classical pathway of
complement activation becoming inoperative. Hence in-
hibitors of proline hydroxylase act as immunosuppres-
sants, for example in Lmmune complex disea~e~.
It is known that the enzyme proline hydroxylase can be
efficiently inhibited by pyridine-2,4- and -2,5-dicar-
boxylic acids (K. Majamaa et al., Eur. J. Biochem. 138
(1984) 239-245). However, in cell culture, these com-
pounds are effective inhibitors only in very high concen-
trations (Tschank, G. et al., Biochem. J. 248 (1987)
2 ~ ~ 2 3~. ~J
- 2 -
625-633).
DE-A 34 32 094 de6cribes pyridine-2,4- and -2,5-dicar~
boxylic diesters with 1-6 carbon atoms in the ester alkyl
moiety as pharmaceuticals for inhibition of proline
hydroxylase and lysin~ hydroxylase.
These lower alkylate~ diesters h~ve the disadvanta~e,
however, that hey are rapidly cleaved in the body to the
acids and do not reach their ~ite of action in the cell
in suf~iciently high concentration and thus are
relatively little suited ~or pos~ible administxation a~
pharmaceuticals~ -
DE-A 37 03 959, DE-A 37 03 952 and DE-~ 37 03 963 de-
scribe in a general form mixed estersJamidas, higher
alkylated diesters and diamides of pyridine-2~4- and
-2,5 dicarbo~ylic acids which are effective inhibitor~ of
collagen biosynthe~is in animal models. Thus~
DE-A 37 03 959 de~cribe~, inter alia, the ~ynthesi~ of
N,N~-bis~2-methoxyethyl)pyridine-2,4-dicarboxamide and
N,N'-bi~(3-i~opropoxypropyl)pyridine-2,4-dicaxhoxamide.
German Paten~ Applications P 38 26 471.4 and
P 38 28 140.6 describe an Lmproved pro~e~s for preparing
N,~-bis(2-methoxyethyl)pyridine-2~4-dicarboxamide.
German Pate~t Application P 39 24 093.2 proposes novel
N,N'-bi~(alkoxyalkyl)pyridine-2,4-dicar~oxamides.
German Patent Application P 40 01 002.3 describes the u~e
of di(nitroxyalkyl)amides of pyridina-2,4- and -~,5-
dicarboxylic acids for preparing pharmaceuticals inhibit-
ing proline hydroxylase and lysine hydroxylase.
Both pyridine-2,4- and -2,5-dicarboxamide (Hirakata
et al.~ J. Pharm. Soc. Japan 77 ~1957) 219 and Haring
et al., Helv. 37 (1954) 147, 153) and pyridine-2,4- and
-2,5-dicarbohydrazide (Itai et al., Bl. Nation. Hyg.
-- 3 --
Lahor. ~okyo, 74 tl956) 115, 117 and Shin~har~ et al.,
Chem. High Polymers Japan, i5 (1~5~) 839) have already
heen disclosed as agents for tuberculosis.
JP 53/28175 (78/28175) de~cribes N,N'-~is(2-nitroxy-
ethyl)pyridlne-2,4- and ~,5 dicarboxEm~des a~ ~ubstances
with a vasodilator actios~.
German Patent Application P .. .. . ~ . (HOE 90/F 192 )
describes the u~e of 2,4- and 2,5-sub~tituted pyridine N-
oxides for preparing pharmaceuticals :inhibiting proline
hydxoxylase and lysine hydroxylase.
It has now been found, surprisingly, that 4- or 5-~ubsti-
tuted pyridine~2-carboxylic acids of the formula I
indicated below/ and the physiologically tolerated ~alts
thereof, effec~ively inhibit lysine hydroxylase and
proline hydroxyla~e in animal models.
Thus the invention claims compounds of the formula I
R1R2N(O)C-
(I)
N COOH
in which
Rl and R2 are, independently of one another, hydrogen,
C1-Cl2-alkyl, C2-Cl2~alkenyl, C2-Cl2-alkynyl, non-benzo-
fused or benzo-~u~ed C5 C7-cycloalkyl, aryl or hetero-
aryl, where these radicals mentioned for R1 and R2 are
unsubstituted or su~stituted by one or more identical
or different radicals R3, where
R3 is halogen, hydroxyl, cyano, nitro, nitroxy, amino,
carboxyl, Cl-C4-alkoxy, Cl-C4-alkoxycarbonyl, C1-C4-
alkyl~ or -dialkylamino, indolyl or phenyl, where the
indolyl and phenyl radical is unsubstituted or substi-
tuted once, twice or three times by halogen, nitro,
C1-C4-alkyl or C1-C4-alkoxy, where in the case o~
3 ~
-- 4 --
multiple sub~titution the radicals are identical or
di~ferent,
or, if
Rl is hydro~en, R2 is a radical -N~R4)(R53 in which
R4 and R5 are identical or different and are ~ydrogen,
C~-Cb-alkyl, C1-3-alkylcar~onyl ox phenyl,
or, if
~1 i8 hydrogen, R2 is a residue of an ~-~mino acid, ~-
amino acid alkyl ester, ~-amino amide, ~-amino alkyl-
or dialkylamide, which i8 bonded via the N terminuæ~
where said al~yl radical~ have 1 to 4 carbon atoms and
are optionally monosubstituted by phenyl, and whare
the C3- and C4~alkyl radical~ can also be branched,
or, if
R1 is hydrogent R2 i~ a residue of a di- or tripeptide
which is bonded via the N terminus, where ~Iresic1ue~
means the particular ~-amino acid and the derivati~e~
thereof or the di- or tripeptide, in each case lacking
the N texminus,
and
Rl has the meaning of R2~ where tha radicalfi R1 and R2 are
identical or different, or
Rl and R2 are, together with the nitrogen at~m to which
they are bo~ded, a radical of the formula II
~ (II)
-N A
(CH2)n
in which
n i~ 1 to 3 and
A is O, S, CH2 or N(R6)-, where
R6 i8 hydrogen, phenyl, C~-C6-alkyl, C2-C6 alkenyl or
C2-C6-alkynyl, where these said radicals are unsub-
stituted or substituted by phenyl which in turn is
unsubstituted or substituted once or more than once
by one or more identical or different substituents
selected from: halogen, nitro, cyano, carboxyl,
- 5 _ 2 ~
hydroxyl, methyl, ethyl, methoxy, ethoxy and
trifluoromethyl,
or
-N(R7)2 where
R7 is hydrogen or Cl-C3-alkyl,
or
-CooR7
or
-CON(Ra)2 or -CONHRB, where
R3 has the meaning of R7 or wherQ (Ra)2 is a C4-C~-
alkylene chain in which zero or one CHa group which
is not directly adjacent to the nltrogen atom i~
replaced by U, S or N-R7,
or where
~ is Cl-C4-alko~ycarbonyl or C3-C7-cycloalkyl,
and the physiologically tolerated 8alt8, where pyridine-
4- and -5-carboxamide-2-caxboxylic acid are excepted.
Particularly preferred compounds of the formula I are
tho~e in which
R1 and R2 are, independently of one another, hydrogen,
Cl-C~;-al3cyl, C2-C6-alkenyl, C2-C6-alkynyl, C5-C7-cyclo-
alkyl, aryl or heteroaryl, where these radicals
mentioned for Rl and R2 are unsubstituted or substi-
tuted by one or two identical or different radi~als
R3, where
R3 is halogen, hydroxyl, cyano, amino, car~oxyl, Cl-C4-
alkoxy, C1-C4-alkoxycarbonyl~ Cl-C4-alkyl- or -dialkyl-
amino, or phenyl~ where the phenyl radical i8 un~ub-
~tituted or ~ubstituted once by halogen, Cl-C2-alkyl or
Cl-C2-alkoxy,
or, if
Rl iS hydrOgell/ R2 i8 a radical -N(R4)(R5~ in which R4 and
R5 are identical or different and are hydrogen or
Cl-C3-alkyl,
or, if
Rl is hydrogen, R2 is a residue of an ~-amino ~cid or ~-
amino acid alkyl ester bonded via the N terminus,
where the alkyl radical has 1 to 3 carbon atoms and is
- 6 ~ ~ ~32
opt~onally monosubstituted hy phenyl, and where the
C3-alkyl radical can al80 b~ branched,
and
Rl ha~ the meaning of R2, where the radicals R1 and R2 are
5identical or dif~erent, or
Rl and R~ are, together with the nitrogen atom to which
they are bonded, a radical of the fonmula II
-N A (I~
(GH2)n
in which
lOn is 1 to 3, and
A is 0, CH2 or -N(R5)-, where
R6 is hydrogen, phenyl, C1-C6-alkyl, where these said
radicals are unsub tituted or substituted by C1-C4-
alkoxycarbonyl or C3-C,-cycloalkyl,
and the physiologically tolerated salts, where pyridine-
4- and -5-caxboxamide-2-carboxylic acid are excepted.
Especially preerrad compounds of the formula I are those
in which
R1 and R2 are, independently of one another, hydro~en,
Cl-C6-alkyl, especially ~-C3-alkyl, C~-cycloal~qyl,
phenyl, or pyridyl, where these radicals mentioned for
Rl and R2 are unsubstituted or ~ubstituted by one or
two identical radicals R3, where
R3 is hydroxyl, amino, carboxylr Cl-C4-alkoxy, especially
methoxy, C1-C4-alkoxycarbonyl, nr phenyl, where the
phenyl radical is unsubstituted or substituted once by
methyl or metho~y,
and
Rl has the meaning of R2/ where the radicals R1 and R2 are
identical or different, or
and R2 are, together with the nitrogen atom to which
they are bonded, a radical of the formula II
(II)
-N A
(CH2)n
in which
n i8 2 and
~ i8 0 or CH2,
5 and the physiologically tolerated salts, where pyrldine-
4- and ~5 carbox~mide-2-carboxylic acid are excepted.
Said compounds pyridine-4- and ~-~-carbox~mide-2-car-
boxylic acid have already been described as such in
Thums, L.J. Pharm. Belg. 24 (1-2), 3~ 69~ and
Delar~e, J.: Pharm. ~cta. Helv. 44 (10), 637-43 (1969).
Halogen means fluorine, chlorine, br~mine and iodine,
aryl means phenyl and naphthyl, and heteroaryl mesns 5-
and 6-member~d aromatic rings with 1, 2 or 3 nitrogen
and~or oxygen and/or sulphur atoms~ which can al~o
optionally be benzo-~used; the he~eroaryl radlcals are,
in particular, pyridyl, pyridazyl, pyrimidyl, pyrazyl,
1,3,5-triazyl, pyrrolyl, pyrazolyl, imidazolyl, tri-
azolyl, tetrazolyl, thienyl, oxazolyl and thia~olyl
radicals and optionally ~hair benzo-fu~ed compounds.
"Substituted more than once`' means herein~efora and
hereinafter that at least 2 and at mo~t all the hydrogen
atoms present in the alkyl radicals are replaced by ~aid
substituents. In this connection there is preferably one
sub6tituent per methyl ~r m~thylene gxoup.
In the ca~e of multiple substitutions, the substituent~
can also be independent of one another.
The invention also relates to the use of compounds of the
formula I and to the physiologically tolerated salts plus
pyridine-4- and -5-carboxamide-2-carboxylic acid for
producinq a pharmaceutical inhibiting proline hydroxylase
and lysin~ hydroxylase.
Finally, the invention relates to ~he compounds of ~he
formula I plus pyridine-4- and -5-carboxamide-2-car~ox-
S ylic acid for use as pharmaceuticals.
The in~ention particularly r21ates to ~he compounds of
the formula I plus pyridine-4- and -5-carboxamide-2-
carboxylic acid for use as fibrosuppressants and immuno-
suppressant~ and for inhibiting proline hydroxylase.and
lysine hydroxylase and for influencing the metaboli~m of
collag~n and collagen-like substances and the biosynthe-
si~ of Clq.
All said alkyl radicals with more than 2 carbon atoms can
be either straight-chain or branched.
lS The invention al~o relates to a process for preparing
compounds of the foxmula I plus pyridine-4- or -5-carbox-
amide-2-carboxylic acids.
The process according to the invention is illustrated
hereinafter taking the example of pyridine 2,4-dicar-
boxylic acid. The process of compounds according to theinven~ion with 2,5-substitution is carried out in an
analogous manner.
2~ . {j
Reaction scheme
23 8 0zCH
Stage 21
~H2~ _ ~H2
CO2CH3 S tage 3 N O~H
¦ Stufe 4
CO2H CONR I R~
~II) e~l C~2C~i3
¦ S t a g e 6
.
CON~1R~
(I')
~02H
In Stage 1, pyridine-2,4~dicarboxy1ic acid, which can be
2t~ ,? 1 1 ~
-- 10 --
bought~ i8 converted into its dicarbonyl dihalide,
preferably it~ dichloride, and reacted with an optionally
substituted benzyl alcohol to give the dibenzyl pyridine-
2,4-dicarboxylate.
In Stage 2, the diester is selectively hydrolyzed in
position 2, for example in th~ presence of a copper
cataly~t as described by Delarge, ~.s Phar. Acta.
H~lv. 44 (10), 637 (1969~.
The free acid functionality in position 2 i5 sub~eguent-
lyr in Stage 3, converted into the corresponding acid
chloride and converted with an ~lcohol ~uch a~, for
example, methyl or ethyl alcohol into the corresponding
2~arbo~ylic e~ter.
~ he remaining ben~yl protective group in positio~ 4 iiB in
Stage 4 eliminated by hydrogenolysis (for example with
H2~Pd, Houben-Weyl: Vol. IV/lc (1980~, pp. 381-82)/ and
the free acid in position 4 is converted into its acid
chloride.
The acid chloride can now be converted with the amine of
the formula III in which Rl and R~ have the meaning a~ in
the compounds of the formula I into the mixed py.ridine-
4-carboxamide-2-carboxylic ester.
~he 2-carboxylic ester can now finally (Sta~e 6) be
converted with alcoholic alkali (for example sodium
hydroxide solution in ethanol) first into tha salt of the
2-carboxylic acid, which is then converted with mineral
acids (such a~, for example, hydrochloric acid) into the
compound of the formula I' according to the invention.
Said process which has been described according to the
reaction scheme for the compounds substituted in
position 4 also applies to the compounds which are
correspondingly substituted in position 5.
~ ~3 $ ~ J
The compounds of tha formula I plus pyridine-4- and -5-
carboxamide-2-carboxylic acid have valuable pharmaco-
logical propertie~ an~ ~how, in particular, activ~ty a~
inhibitor~ o~ ~roline hydroxylase and lysine hydroxylase,
as fibrosuppres~ant and immunosuppressant.
The activity of the ~ibrogenase can ~e de~e~mined by
radioimmunological determination of the N-terminal
propeptide of collagen type III or of the N- or C-
terminal cros~linking domain of collagen typa IV t7s
collagen or type I~ collagen ~Cl) in ~erum.
For this purpose, the hydroxyproline, procollagen III
peptide~ 7s collagen and type IV collagen NCl concentra-
tion~ were measured in the liver of
a~ untreated rats ~control)
b) rats given tetrachloromethane ( CC14 cont~ol3
c) rats given first CC14 and then a compound according to
the invention
(this test method is described by Rouiller, C., experi-
mental toxic in~ury o~ the liver; in The Liver,
2~ C. Rouiller, Vol. 2, pp. 335-476, New York, Academic
Press, 1964).
The compounds of the fonmula I plus pyridina-4- and -5-
carboxamide 2-carboxylic acid can be used as medicament~
in the form of pharmaceutical products which contain them
where appropriate together with tolerated pharmaceutical
vehicles. The compounds can be used as medi~ine~, ~or
example in the form of pharmaceutical products which
contain these compounds mixed with a pharmaceutical,
organic or inorganic vehicle suitable for enteral,
percutaneous or parenteral administration, such as~ for
example, water, gum arabic, gelatin, lactose, ~tarch,
magnesium stearate, talc, vegetable oils, polyalkylene
glycols, vaseline etc.
They can be administered for this purpo6e orally in dose6
of 0.01 - 25.0 mg/kg/day, preferably 0.01 - 5.0 mg/kg/day
. Y.
- 12 -
or parenterally in doses of 0~001 - 5 mg/kg/day, prefer-
ably 0.001 - 2.5 mg/kg~day, ~pecially 0.005
1.0 mg/kg/day. It is also possible to increase the dosage
in sevexe ca~eR. However, lower dose~ also 6uffice in
S many ca~es. These data relate to an adult weighing about
75 ~g.
The invention also embraces the use of the compounds
according to the invention ~or preparing pharmaceuticals
which are employed for ~he treatment and prophylaxis of
the abovementioned metabolic disorders.
The invention additionally relates to pharmaceuticals
which contain one or more compounds of the formula I
according to the invPntîon and/or their physiologically
tolerated salts, plus the pyridine-4- and -S-carboxam:ide-
2-carboxylic acids.
The pharmaceuticals are produced by processes Xnown per
se and familiar to the person skilled iYl the art. As
pharmaceuticals, the pharmacologically active compounds
(= active ^~ubstance) according to the invention are
employed either as such or, preferably, in combinati~n
with suitable pharmaceutical auxiliaries or excipients i~
the form of tablets, coated tablets, capsules, supposit-
ories, emulsions, suspen~ions or solutions, where the
content of acti~e substance is up to about 95 %, advant-
ageously between 10 and 75 %~
Suitable auxiliaries and excipients for the reguiredpharmaceutical formulation are, for example, besides
solvents, gel former~, suppository bases, tableting
auxiliaries and other active substance vehicles, also
antioxidants, disper6ants, emulsifier6, foam suppres-
sants, flavorin~s, preservativeæ, solubilizers or
colorants.
The active substances can be administered orally, parent-
erally or rectally.
r ~ r J 3
- 13 -
The active compounds are mixed with the additives ~uit-
able for this r such as excipients, stabilizers or inert
diluents, and converted by the usual methods into ~uit-
able dosage forms such as tablets, coated tablets, hard
S gelatin capsulesl aqueou~ alcoholic or oily suspen~ions
or aqueous or oily solutions.
Examples of inert excipients which can be used are ~um
arabicJ magnesia, magnesium carbonate, potassium pho~-
phate, lactose, glucose or starch, especially corn
starch. The preparation can be carried out either a~ dry
or as wet granules. ~xamples of suitable oily excip~ents
or solvents are vegetable or animal oils~ ~uch a8 ~un-
flower oil or fish liver oil.
For subcutaneous or intravenous administration, the
active compounds are, if required, converted into a
solution, suspQnsion or emulsion with the sub~tances
suitable for this purpose, such as solubilizers, emulsi-
fiers or other auxiliaries~ Examples of suitable solvents
are physiological saline or alcohols, for example eth-
ano~, propanol, glycerol, as well as ~ugar solutions ~uchas glucose or mannitol solutions, or else a mixture of
the various solvents mentioned.
The invention is explained in ~ore de~ail hereinafter by
means of examples.
Example 1
Dibenæyl pyridine-2~4-dicarboxylate (Stage 1)
30 g of pyridine-2t4-dicarboxylic acid are converted with
30 ml of thionyl chloride into the acid chloride and
reacted with 43.8 g of ben%yl alcohol. The product i~
recrystallized from diisopropyl ether.
Yield: 42.1 g melting point 63 - 65C
~ A~ $ S) 1-,`; A~, r3
- 14 -
Example 2
Pyridine-2,4-dicarboxylic acid 4-benzyl ester (Stage 2)
40 g of dibenzyl pyridina-~,4-~icarboxylate from Ex-
ample 1 are ad~ed to a suspension of 27.8 g of copper(II)
nitrate in 700 ml of methanol. ~he mixture is ~o~led
under reflux for one hour and, after cooling, the copper
complex is remo~ed by filtra~ion. The complex is ~uspend
ed in dioxane, and hydrogen sulfide is pa~sed in. The
precipitated coppex sulide is filtered off and the
organic phase is concentrated. The product is stirred
with petroleum ether.
Yield: 25.3 g melting point 113 - 115C
Example 3
4-Benzyl 2-methyl pyridine-2,4-dicarboxylate (Stage 3)
34 g of the compound from Example 2 are introduced into
l l of tetrah~drofuran. The mixt~re is cooled to ODC and,
at this temperature, 250 ml of a diazomethane ~olution
are 910~1y added dropwise. The mixture i allowed to
reach room temperature and is stirred for 12 hours. The
20 solution is concentrated and the crude product is chroma-
tographed (silica g~l, mobile pha~e ethyl acetate).
Yield: 26 g melting point: 58DC
Exampl~ 4
Pyridine-2,4-dicarboxylic a~id 2-methyl ester (Stage 4~
26 g of the compound from Example 3 are dissolved in 1 l
of dioxane at room temperature. 500 mg of palladium/car-
bon (10 ~) catalyst are added to the solution, and
hydrogenation is carried out under atmospheric pressure
for 4 hours. After hydrogen uptake ceases, the catalyst
is filtered off with suction and the solvent iB ~tripped
off~
- 15 -
Yield: 15.3 g melting point 241 - 243C
Example 5a
Pyridine-2,4-dicarboxylic acid 2-methyl es~er 4-(2-
methoxyethyl)amide ~Staqe 5)
ll g of the compound from Example 4 are converted in
analogy to Example 1 with 4.8 ml o~ thionyl chloride into
the acid chloride and reacted with S.~ ml of 2-methoxy-
ethylamine to give the amide. The crude product is
triturated with diisopropyl ether and filtered off with
suction.
Yield: 9.5 g melting point: 74 - 76C
Example 5b
Pyridine-2~4-dicarboxylic acid 2-methyl ester 4-(3-
mekhoxypropyl)amide ~Stage S)
In analogy to Example 5a, 3 g of the compound from
Example 4 are converted with 1.2 ml of thionyl chloride
into the acid chloride and reacted with 1.7 ml o 3-
methoxypropylamine to give the amide. ~he crude product
i8 chromatographed (ethyl acetateJmethanol 15~1).
Yield: 2.6 g oil
Example 6a
Pyridine-2,4-dicarboxylic acid 4-(2-methoxyethyl)amide
(Stage 6)
2.3 g of sodium hydro~ide ~pellets) Are dissolved in
70 ml of ethanol at room temperature. A solution of 9 g
of the compound from Example 5a in 30 ml of ethanol is
added dropwise at this temperature. After 4 hours, the
mixture is evaporated to dryness, the residue is taken up
in a little water and the solution is slowly acidified
with concentrated hydrochloric acid. The solution i3
~ ~ è3 ~ ~? ,3 ,~i
16 -
evaporated and the product is recrys~allized from hot
ethanol.
Yield: 8 g melting point~ 130 - 135C
Example 6b
Pyridine-~,4-dicarboxylic acid 4-(3-methoxypropyl)ami~e
(Stage 6)
The compou~d is prepared in analogy to ~xample Xa from
3 g of the e~er fr~m Example 5b.
Yield: 2.6 g melting point: 118 - 1224C
