U.S. patent application number 14/996678 was filed with the patent office on 2016-05-12 for substituted 5-fluoro-1h-pyrazolopyridines and their use.
This patent application is currently assigned to ADVERIO PHARMA GMBH. The applicant listed for this patent is ADVERIO PHARMA GMBH. Invention is credited to Jens ACKERSTAFF, Eva-Maria BECKER, Donald BIERER, Markus FOLLMANN, Nils GRIEBENOW, Elke HARTMANN, Rolf JAUTELAT, Andreas KNORR, Walter KROH, Volkhart Min-Jian Li, Joachim MITTENDORF, Gorden REDLICH, Karl-Heinz SCHLEMMER, Johannes-Peter STASCH, Frank WUNDER.
Application Number | 20160129004 14/996678 |
Document ID | / |
Family ID | 44119218 |
Filed Date | 2016-05-12 |
United States Patent
Application |
20160129004 |
Kind Code |
A1 |
FOLLMANN; Markus ; et
al. |
May 12, 2016 |
SUBSTITUTED 5-FLUORO-1H-PYRAZOLOPYRIDINES AND THEIR USE
Abstract
The present application relates to novel substituted
5-fluoro-1H-pyrazolopyridines, to processes for their preparation,
to their use alone or in combinations for the treatment and/or
prophylaxis of diseases, and to their use for producing medicaments
for the treatment and/or prophylaxis of diseases, in particular for
the treatment and/or prophylaxis of cardiovascular disorders.
Inventors: |
FOLLMANN; Markus; (Cologne,
DE) ; STASCH; Johannes-Peter; (Solingen, DE) ;
REDLICH; Gorden; (Bochum, DE) ; ACKERSTAFF; Jens;
(Dusseldorf, DE) ; GRIEBENOW; Nils; (Dormagen,
DE) ; KROH; Walter; (Wuppertal, DE) ; KNORR;
Andreas; (Erkrath, DE) ; BECKER; Eva-Maria;
(Wuppertal, DE) ; WUNDER; Frank; (Wuppertal,
DE) ; Li; Volkhart Min-Jian; (Velbert, DE) ;
HARTMANN; Elke; (Wuppertal, DE) ; MITTENDORF;
Joachim; (Wuppertal, DE) ; SCHLEMMER; Karl-Heinz;
(Wuppertal, DE) ; JAUTELAT; Rolf; (Haan, DE)
; BIERER; Donald; (Haan, DE) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
ADVERIO PHARMA GMBH |
Schoenefeld |
|
DE |
|
|
Assignee: |
ADVERIO PHARMA GMBH
Schoenefeld
DE
|
Family ID: |
44119218 |
Appl. No.: |
14/996678 |
Filed: |
January 15, 2016 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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14552122 |
Nov 24, 2014 |
9266885 |
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14996678 |
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13851373 |
Mar 27, 2013 |
8921377 |
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14552122 |
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13111856 |
May 19, 2011 |
8420656 |
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13851373 |
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Current U.S.
Class: |
514/256 ;
544/322 |
Current CPC
Class: |
A61P 7/00 20180101; A61P
9/10 20180101; A61P 9/04 20180101; C07C 309/04 20130101; C07C
309/05 20130101; A61P 11/00 20180101; C07C 57/145 20130101; A61P
9/12 20180101; A61P 9/14 20180101; A61P 15/00 20180101; A61K 31/506
20130101; A61K 45/06 20130101; A61P 13/12 20180101; C07D 471/04
20130101; A61P 9/00 20180101; A61P 7/02 20180101 |
International
Class: |
A61K 31/506 20060101
A61K031/506; A61K 45/06 20060101 A61K045/06; C07C 309/05 20060101
C07C309/05; C07C 57/145 20060101 C07C057/145; C07D 471/04 20060101
C07D471/04; C07C 309/04 20060101 C07C309/04 |
Foreign Application Data
Date |
Code |
Application Number |
May 26, 2010 |
DE |
102010021637.2 |
Claims
1. A compound of the formula (I) ##STR00044## in which R.sup.1
represents hydrogen or (C.sub.1-C.sub.4)-alkyl, where
(C.sub.1-C.sub.4)-alkyl may be substituted by one or two
substituents independently of one another selected from the group
consisting of fluorine and trifluoromethyl, and N-oxides, salts,
solvate salts of the N-oxides and solvates of the N-oxides and
salts thereof.
2. The compound claim 1 in which R.sup.1 represents hydrogen or
methyl, where methyl may be substituted by a trifluormethyl
substitutent, and salts, solvates and solvates of the salts
thereof.
3. (canceled)
4. (canceled)
5. (canceled)
6. (canceled)
7. (canceled)
8. A medicament comprising the compound of claim 1 in combination
with an inert, non-toxic, pharmaceutically suitable excipient.
9. A medicament comprising the compound of claim 1 in combination
with a further active compound selected, from the group consisting
of organic nitrates, NO donors, cGMP-PDE inhibitors, agents having
antithrombotic activity, agents lowering blood pressure, and agents
altering lipid metabolism.
10. (canceled)
11. (canceled)
12. A method of treating or preventing heart failure, angina
pectoris, hypertension, pulmonary hypertension, ischaemias,
vascular disorders, kidney failure, thromboembolic disorders,
fibrotic disorders and arteriosclerosis comprising administering an
effective amount of the compound of claim 1 to a human or animal in
need thereof.
Description
[0001] The present application relates to novel substituted
5-fluoro-1H-pyrazolopyridines, to processes for their preparation,
to their use alone or in combinations for the treatment and/or
prophylaxis of diseases, and to their use for producing medicaments
for the treatment and/or prophylaxis of diseases, in particular for
the treatment and/or prophylaxis of cardiovascular disorders.
[0002] One of the most important cellular transmission systems in
mammalian cells is cyclic guanosine monophosphate (cGMP). Together
with nitrogen monoxide (NO), which is released from the endothelium
and transmits hormonal and mechanical signals, it forms the NO/cGMP
system. Guanylate cyclases catalyze the biosynthesis of cGMP from
guanosine triphosphate (GTP). The known representatives of this
family can be classified both according to structural features and
according to the type of ligands into two groups: the particulate
guanylate cyclases which can be stimulated by natriuretic peptides,
and the soluble guanylate cyclases which can be stimulated by NO.
The soluble guanylate cyclases consist of two subunits and very
probably contain one haem per heterodimer, which is part of the
regulatory site. It is of central importance for the activation
mechanism. NO is able to bind to the iron atom of haem and thus
markedly increase the activity of the enzyme. Haem-free
preparations cannot, by contrast, be stimulated by NO. Carbon
monoxide (CO) is also able to attach to the central iron atom of
haem, but the stimulation by CO is distinctly less than that by
NO.
[0003] By forming cGMP, and owing to the resulting regulation of
phosphodiesterases, ion channels and protein kinases, guanylate
cyclase plays an important role in various physiological processes,
in particular in the relaxation and proliferation of smooth muscle
cells, in platelet aggregation and platelet adhesion and in
neuronal signal transmission, and also in disorders which are based
on a disturbance of the abovementioned processes. Under
pathophysiological conditions, the NO/cGMP system may be
suppressed, which may lead for example to high blood pressure,
platelet activation, increased cellular proliferation, endothelial
dysfunction, atherosclerosis, angina pectoris, heart failure,
myocardial infarction, thromboses, stroke and sexual
dysfunction.
[0004] Owing to the expected high efficiency and few side effects,
a treatment of such disorders which targets the influence of the
cGMP signal path in organisms and is NO-independent is a promising
approach.
[0005] Hitherto, for the therapeutic stimulation of the soluble
guanylate cyclase use has exclusively been made of compounds such
as organic nitrates whose effect is based on NO. This is formed by
bioconversion and activates soluble guanylate cyclase by attack at
the central iron atom of haem. In addition to the side effects, the
development of tolerance is one of the decisive disadvantages of
this type of treatment.
[0006] In recent years, some substances have been described which
stimulate soluble guanylate cyclase directly, i.e. without prior
release of NO, such as, for example
3-(5'-hydroxymethyl-2'-furyl)-1-benzylindazole [YC-1; Wu et al.,
Blood 84 (1994), 4226; Mulsch et al., Brit. J. Pharmacol. 120
(1997), 681], fatty acids [Goldberg et al., J. Biol. Chem. 252
(1977), 1279], diphenyliodonium hexafluorophosphate [Pettibone et
al., Eur. J. Pharmacol. 116 (1985), 307], isoliquiritigenin [Yu et
al., Brit. J. Pharmacol. 114 (1995), 1587] and various substituted
pyrazole derivatives (WO 98/16223).
[0007] WO 00/06569 discloses fused pyrazole derivatives and WO
03/095451 carbamate-substituted 3-pyrimidinylyrazolopyridines as
stimulators of soluble guanylate cyclase.
[0008] It is an object of the present invention to provide novel
substances which act as stimulators of soluble guanylate cyclase
and which have an identical or improved therapeutic profile
compared to compounds known from the prior art, such as, for
example, with respect to their in vivo properties such as, for
example, their pharmacokinetic and pharmacodynamic behaviour and/or
their dose-activity relationship.
[0009] The present invention provides compounds of the general
formula (1)
##STR00001##
in which [0010] R.sup.1 represents hydrogen or
(C.sub.1-C.sub.4)-alkyl, [0011] where (C.sub.1-C.sub.4)-alkyl may
be substituted by one or two substituents independently of one
another selected from the group consisting of fluorine and
trifluoromethyl, and their N-oxides, salts, solvates, salts of the
N-oxides and solvates of the N-oxides and salts.
[0012] Compounds according to the invention are the compounds of
the formula (I) and the N-oxides, salts, solvates and solvates of
the N-oxides and salts thereof, the compounds, encompassed by
formula (I), of the formulae specified hereinafter and the
N-oxides, salts, solvates and solvates of the N-oxides and salts
thereof, and the compounds encompassed by formula (I) and specified
hereinafter as working examples and the N-oxides, salts, solvates
and solvates of the N-oxides and salts thereof, to the extent that
the compounds encompassed by formula (I) and specified hereinafter
are not already N-oxides, salts, solvates and solvates of the
N-oxides and salts.
[0013] In the context of the present invention, preferred salts are
physiologically acceptable salts of the compounds according to the
invention. Also encompassed are salts which are not themselves
suitable for pharmaceutical applications but can be used, for
example, for isolation or purification of the compounds according
to the invention.
[0014] Physiologically acceptable salts of the compounds according
to the invention include acid addition salts of mineral acids,
carboxylic acids and sulphonic acids, for example salts of
hydrochloric acid, hydrobromic acid, sulphuric acid, phosphoric
acid, methanesulphonic acid, ethanesulphonic acid, toluenesulphonic
acid, benzenesulphonic acid, naphthalenedisulphonic acid, formic
acid, acetic acid, trifluoroacetic acid, propionic acid, lactic
acid, tartaric acid, malic acid, citric acid, fumaric acid, maleic
acid and benzoic acid.
[0015] Physiologically acceptable salts of the compounds according
to the invention also include salts of conventional bases, by way
of example and with preference alkali metal salts (e.g. sodium and
potassium salts), alkaline earth metal salts (e.g. calcium and
magnesium salts) and ammonium salts derived from ammonia or organic
amines having 1 to 16 carbon atoms, by way of example and with
preference ethylamine, diethylamine, triethylamine,
ethyldiisopropylamine, monoethanolamine, diethanolamine,
triethanolamine, dicyclohexylamine, dimethylaminoethanol, procaine,
dibenzylamine, N-methylmorpholine, arginine, lysine,
ethylenediamine and N-methylpiperidine.
[0016] In the context of the invention, solvates refer to those
forms of the compounds according to the invention which, in the
solid or liquid state, form a complex by coordination with solvent
molecules. Hydrates are a specific form of the solvates in which
the coordination is with water. Preferred solvates in the context
of the present invention are hydrates.
[0017] Depending on their structure, the compounds according to the
invention may exist in different stereoisomeric forms, i.e. in the
form of configurational isomers or if appropriate also as
conformational isomers (enantiomers and/or diastereomers, including
those in the case of atropisomers). The present invention therefore
encompasses the enantiomers or diastereomers and the respective
mixtures thereof. The stereoisomerically uniform constituents can
be isolated from such mixtures of enantiomers and/or diastereomers
in a known manner; chromatography processes are preferably used for
this, in particular HPLC chromatography on an achiral or chiral
phase.
[0018] Where the compounds according to the invention can occur in
tautomeric forms, the present invention encompasses all the
tautomeric forms.
[0019] The present invention also encompasses all suitable isotopic
variants of the compounds according to the invention. An isotopic
variant of a compound according to the invention is understood here
to mean a compound in which at least one atom within the compound
according to the invention compound has been exchanged for another
atom of the same atomic number, but with a different atomic mass
than the atomic mass which usually or predominantly occurs in
nature. Examples of isotopes which can be incorporated into a
compound according to the invention are those of hydrogen, carbon,
nitrogen, oxygen, phosphorus, sulphur, fluorine, chlorine, bromine
and iodine, such as .sup.2H (deuterium), .sup.3H (tritium),
.sup.13C, .sup.14C, .sup.15N, .sup.17O, .sup.18O, .sup.32P,
.sup.33P, .sup.33S, .sup.34S, .sup.35S, .sup.36S, .sup.18F,
.sup.36Cl, .sup.82Br, .sup.123I, .sup.124I, .sup.129I and
.sup.131I. Particular isotopic variants of a compound according to
the invention, especially those in which one or more radioactive
isotopes have been incorporated, may be beneficial, for example,
for the examination of the mechanism of action or of the active
compound distribution in the body; due to comparatively easy
preparability and detectability, especially compounds labelled with
.sup.3H or .sup.14C isotopes are suitable for this purpose. In
addition, the incorporation of isotopes, for example of deuterium,
can lead to particular therapeutic benefits as a consequence of
greater metabolic stability of the compound, for example an
extension of the half-life in the body or a reduction in the active
dose required; such modifications of the compounds according to the
invention may therefore in some cases also constitute a preferred
embodiment of the present invention. Isotopic variants of the
compounds according to the invention can be prepared by processes
known to those skilled in the art, for example by the methods
described below and the methods described in the working examples,
by using corresponding isotopic modifications of the particular
reagents and/or starting compounds therein.
[0020] Moreover, the present invention also encompasses prodrugs of
the compounds according to the invention. Here, the term "prodrugs"
refers to compounds which for their part can be biologically active
or inactive, but are converted (for example metabolically or
hydrolytically) into compounds according to the invention during
their dwell time in the body.
[0021] In the context of the present invention, unless specified
otherwise, the substituents are defined as follows:
[0022] In the context of the invention, alkyl represents a
straight-chain or branched alkyl radical having 1 to 4 carbon
atoms. The following may be mentioned by way of example and by way
of preference: methyl, ethyl, n-propyl, isopropyl, n-butyl,
isobutyl, 1-methylpropyl, tert-butyl.
[0023] In the context of the invention, halogen represents
fluorine, chlorine, bromine and iodine.
[0024] If radicals in the compounds according to the invention are
substituted, the radicals may be mono- or polysubstituted, unless
specified otherwise. In the context of the present invention, all
radicals which occur more than once are defined independently of
one another. Substitution by one, two or three identical or
different substituents is preferred.
[0025] In the context of the present invention, preference is given
to compounds of the formula (I) in which [0026] R.sup.1 represents
hydrogen or methyl, [0027] where methyl may be substituted by a
trifluoromethyl substituent, and their salts, solvates and solvates
of the salts.
[0028] In the context of the present invention, particular
preference is given to the following compounds of the formula (I):
[0029] methyl
{4,6-diamino-2-[5-fluoro-1-(2-fluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-y-
l]pyrimidin-5-yl}carbamate [0030] methyl
{4,6-diamino-2-[5-fluoro-1-(2-fluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-y-
l]pyrimidin-5-yl}methylcarbamate [0031] methyl
{4,6-diamino-2-[5-fluoro-1-(2-fluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-y-
l]pyrimidin-5-yl}(2,2,2-trifluoroethyl)carbamate [0032] methyl
{4,6-diamino-2-[5-fluoro-1-(2-fluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-y-
l]pyrimidin-5-yl}carbamate hydrochloride [0033] methyl
{4,6-diamino-2-[5-fluoro-1-(2-fluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-y-
l]pyrimidin-5-yl}carbamate sulphate [0034] methyl
{4,6-diamino-2-[5-fluoro-1-(2-fluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-y-
l]pyrimidin-5-yl}carbamate phosphate [0035] methyl
{4,6-diamino-2-[5-fluoro-1-(2-fluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-y-
l]pyrimidin-5-yl}carbamate mesylate [0036] methyl
{4,6-diamino-2-[5-fluoro-1-(2-fluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-y-
l]pyrimidin-5-yl}carbamate ethane-1,2-disulphonate [0037] methyl
{4,6-diamino-2-[5-fluoro-1-(2-fluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-y-
l]pyrimidin-5-yl}carbamate maleate [0038] methyl
{4,6-diamino-2-[5-fluoro-1-(2-fluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-y-
l]pyrimidin-5-yl}carbamate nitrate
[0039] The individual radical definitions specified in the
respective combinations or preferred combinations of radicals are,
independently of the respective combinations of the radicals
specified, also replaced as desired by radical definitions of other
combinations.
[0040] Particular preference is given to combinations of two or
more of the preferred ranges mentioned above.
[0041] The invention further provides a process for preparing the
compounds of the formula (I) according to the invention,
characterized in that the compound of the formula (II)
##STR00002## [0042] [A] is reacted in an inert solvent in the
presence of hexabutyltin and a suitable palladium catalyst with
intermediate formation of a tin species with the compound of the
formula (III)
[0042] ##STR00003## [0043] to give the compound of the formula
(IV)
[0043] ##STR00004## [0044] this is then reduced in an inert solvent
with a suitable reducing agent to give the compound of the formula
(V)
[0044] ##STR00005## [0045] and this is then reacted in the presence
of a suitable base in the presence or absence of a solvent with
methyl chloroformate to give the compound of the formula (I-A)
##STR00006##
[0045] or [0046] [B] the compound of the formula (II) is reacted in
an inert solvent with copper cyanide to give the compound of the
formula (VI)
[0046] ##STR00007## [0047] this is then, under acidic conditions,
converted into the compound of the formula (VII)
[0047] ##STR00008## [0048] this is subsequently reacted in an inert
solvent in the presence of a suitable base with the compound of the
formula (VIII)
[0048] ##STR00009## [0049] to give the compound of the formula
(IX)
[0049] ##STR00010## [0050] and this is then reduced in an inert
solvent in the presence of a suitable reducing agent to give the
compound (V), and this is subsequently reacted further according to
process [A] to give compound (I-A), or [0051] [C] the compound of
the formula (I-A) is reacted in an inert solvent in the presence of
a suitable base with a compound of the formula (X)
[0051] R.sup.1A--X.sup.1 (X),
in which [0052] R.sup.1A represents (C.sub.1-C.sub.4)-alkyl, [0053]
where (C.sub.1-C.sub.4)-alkyl may be substituted by one or two
substituents independently of one another selected from the group
consisting of fluorine and trifluoromethyl, [0054] and [0055]
X.sup.1 represents a leaving group such as, for example, halogen,
in particular bromine or iodine, trichloromethanesulphonate,
mesylate or tosylate [0056] to give a compound of the formula
(I-B)
[0056] ##STR00011## [0057] in which R.sup.1A has the meaning given
above, and the resulting compounds of the formulae (I-A) and (I-B)
are, where appropriate, converted with the appropriate (i) solvents
and/or (ii) acids or bases into their solvates, salts and/or
solvates of the salts.
[0058] Inert solvents for process step (II)+(III).fwdarw.(IV) are,
for example, alcohols such as methanol, ethanol, n-propanol,
isopropanol, n-butanol or tert-butanol, ethers such as diethyl
ether, dioxane, tetrahydrofuran, glycol dimethyl ether or
diethylene glycol dimethyl ether, hydrocarbons such as benzene,
xylene, toluene, hexane, cyclohexane or mineral oil fractions, or
other solvents such as dimethylformamide (DMF), dimethyl sulphoxide
(DMSO), N,N'-dimethylpropyleneurea (DMPU), dimethylacetamide,
N-methylpyrrolidone (NMP), pyridine, acetonitrile, sulpholane or
else water. It is equally possible to use mixtures of the solvents
mentioned. Preference is given to dioxane.
[0059] Suitable palladium catalysts for process step
(II)+(III).fwdarw.(IV) are, for example, palladium on activated
carbon, palladium(II) acetate,
tetrakis(triphenylphosphine)palladium(0),
bis(triphenylphosphine)palladium(II) chloride,
bis(acetonitrile)palladium(II) chloride and
[1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II)/dichlorometha-
ne complex, if appropriate in combination with additional phosphane
ligands such as, for example, (2-biphenyl)di-tert-butylphosphine,
dicyclohexyl[2',4',6'-tris(1-methylethyl)biphenyl-2-yl]phosphane
(XPHOS), bis(2-phenylphosphinophenyl) ether (DPEphos) or
4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (Xantphos) [cf.,
for example, Hassan J. et al., Chem. Rev. 102, 1359-1469 (2002)].
Preference is given to using
tetrakis(triphenylphosphine)palladium(0).
[0060] The reaction (II)+(III).fwdarw.(IV) is generally carried out
in a temperature range of from +20.degree. C. to +180.degree. C.,
preferably from +50.degree. C. to +120.degree. C., if appropriate
in a microwave. The reaction can be carried out at atmospheric,
elevated or reduced pressure (for example from 0.5 to 5 bar). The
reaction is generally carried out at atmospheric pressure.
[0061] The reductions (IV).fwdarw.(V) and (IX).fwdarw.(V) are
carried out in the presence of a suitable catalyst in an inert
solvent in a temperature range of from +20.degree. C. to
+40.degree. C. under hydrogen of atmospheric pressure.
[0062] Inert solvents for the reductions (IV).fwdarw.(V) and
(IX).fwdarw.(V) are, for example, alcohols such as methanol,
ethanol, n-propanol, isopropanol, n-butanol or tert-butanol, ethers
such as diethyl ether, dioxane, tetrahydrofuran, glycol dimethyl
ether or diethylene glycol dimethyl ether, or other solvents such
as dimethylformamide (DMF), dimethyl sulphoxide (DMSO),
N',N'-dimethylpropyleneurea (DMPU), N-methylpyrrolidone (NMP),
pyridine, acetonitrile or else water. It is equally possible to use
mixtures of the solvents mentioned. Preference is given to DMF and
pyridine.
[0063] Suitable catalysts for the reactions (IV).fwdarw.(V) and
(IX).fwdarw.(V) are, for example, palladium on activated carbon,
platinum on carbon, palladium hydroxide or Raney nickel.
[0064] Alternatively, the reductions (IV).fwdarw.(V) and
(IX).fwdarw.(V) can be carried out using a metal or metal salt such
as, for example, iron, zinc or tin(II) chloride in a suitable acid
such as, for example, hydrogen chloride/hydrochloric acid,
sulphuric acid, phosphoric acid or acetic acid in a temperature
range of from +20.degree. C. to +140.degree. C.
[0065] Inert solvents for process step (V).fwdarw.(I-A) are, for
example, alcohols such as methanol, ethanol, n-propanol,
isopropanol, n-butanol or tert-butanol, ethers such as diethyl
ether, dioxane, tetrahydrofuran, glycol dimethyl ether or
diethylene glycol dimethyl ether, halogenated hydrocarbons such as
dichloromethane, trichloromethane, carbon tetrachloride,
trichloroethylene or chlorobenzene, hydrocarbons such as benzene,
xylene, toluene, hexane, cyclohexane or mineral oil fractions, or
other solvents such as dimethylformamide (DMF), dimethyl sulphoxide
(DMSO), N,N'-dimethylpropyleneurea (DMPU), N-methylpyrrolidone
(NMP), acetonitrile or else water. It is equally possible to use
mixtures of the solvents mentioned. Preference is given to
dimethylformamide and toluene and also to a mixture of
dimethylformamide and toluene.
[0066] Suitable bases for the process step (V).fwdarw.(I-A) are
alkali metal hydrides such as sodium hydride, alkali metal
hydroxides such as, for example, lithium hydroxide, sodium
hydroxide or potassium hydroxide, alkali metal carbonates such as
lithium carbonate, sodium carbonate, potassium carbonate or caesium
carbonate, alkali metal bicarbonates such as sodium bicarbonate or
potassium bicarbonate, alkali metal alkoxides such as sodium
methoxide or potassium methoxide, sodium ethoxide or potassium
ethoxide or potassium tert-butoxide, or organic amines such as
triethylamine, diisopropylethylamine, pyridine,
1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) or
1,5-diazabicyclo[4.3.0]non-5-ene (DBN). Preference is given to
pyridine.
[0067] The reaction (V).fwdarw.(I-A) is generally carried out in a
temperature range of from -10.degree. C. to +30.degree. C.,
preferably from 0.degree. C. to +20.degree. C. The reaction can be
carried out at atmospheric, elevated or reduced pressure (for
example from 0.5 to 5 bar). The reaction is generally carried out
at atmospheric pressure.
[0068] Inert solvents for process step (II).fwdarw.(VI) are, for
example, ethers such as diethyl ether, dioxane, tetrahydrofuran,
glycol dimethyl ether or diethylene glycol dimethyl ether,
hydrocarbons such as benzene, xylene, toluene, hexane, cyclohexane
or mineral oil fractions, or other solvents such as
dimethylformamide (DMF), dimethyl sulphoxide (DMSO),
N,N'-dimethylpropyleneurea (DMPU), N-methylpyrrolidone (NMP),
pyridine or acetonitrile. It is equally possible to use mixtures of
the solvents mentioned. Preference is given to DMSO.
[0069] The reaction (II).fwdarw.(VI) is generally carried out in a
temperature range of from +20.degree. C. to +180.degree. C.,
preferably from +100.degree. C. to +160.degree. C., if appropriate
in a microwave. The reaction can be carried out at atmospheric,
elevated or reduced pressure (for example from 0.5 to 5 bar). The
reaction is generally carried out at atmospheric pressure.
[0070] The reaction (VI).fwdarw.(VII) is carried out using methods
known to the person skilled in the art in a two-step process
initially with formation of the imino ester using sodium methoxide
in methanol at from 0.degree. C. to +40.degree. C. and subsequent
nucleophilic addition of an ammonia equivalent such as, for
example, ammonia or ammonium chloride in acetic acid with formation
of the amidine (VII) at from +50 to +150.degree. C.
[0071] Inert solvents for process step (VII)+(VIII).fwdarw.(IX) are
alcohols such as methanol, ethanol, n-propanol, isopropanol,
n-butanol or tert-butanol, ethers such as diethyl ether, dioxane,
tetrahydrofuran, glycol dimethyl ether or diethylene glycol
dimethyl ether, hydrocarbons such as benzene, xylene, toluene,
hexane, cyclohexane or mineral oil fractions, or other solvents
such as dimethylformamide (DMF), dimethyl sulphoxide (DMSO),
N',N'-dimethylpropyleneurea (DMPU), N-methylpyrrolidone (NMP),
pyridine, acetonitrile or else water. It is equally possible to use
mixtures of the solvents mentioned. Preference is given to DMF.
[0072] Suitable bases for the process step (VII)+(VIII).fwdarw.(IX)
are alkali metal hydroxides such as, for example, lithium
hydroxide, sodium hydroxide or potassium hydroxide, alkali metal
carbonates such as lithium carbonate, sodium carbonate, potassium
carbonate or caesium carbonate, alkali metal bicarbonates such as
sodium bicarbonate or potassium bicarbonate, alkali metal alkoxides
such as sodium methoxide or potassium methoxide, sodium ethoxide or
potassium ethoxide or potassium tert-butoxide, or organic amines
such as triethylamine, diisopropylethylamine, pyridine,
1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) or
1,5-diazabicyclo[4.3.0]non-5-ene (DBN). Preference is given to
triethylamine.
[0073] The reaction (VII)+(VIII).fwdarw.(IX) is generally carried
out in a temperature range of from +20.degree. C. to +150.degree.
C., preferably from +80.degree. C. to +120.degree. C., if
appropriate in a microwave. The reaction can be carried out at
atmospheric, elevated or reduced pressure (for example from 0.5 to
5 bar). The reaction is generally carried out at atmospheric
pressure.
[0074] The compound of the formula (VIII) can be prepared
analogously to the literature L. F. Cavalieri, J. F. Tanker, A.
Bendich, J. Am. Chem. Soc., 1949, 71, 533.
[0075] Inert solvents for the reaction (I-A).fwdarw.(I-B) are, for
example, halogenated hydrocarbons such as dichloromethane,
trichloromethane, carbon tetrachloride, trichloroethylene or
chlorobenzene, ethers such as diethyl ether, dioxane,
tetrahydrofuran, glycol dimethyl ether or diethylene glycol
dimethyl ether, or other solvents such as dimethylformamide (DMF),
dimethyl sulphoxide (DMSO), N,N'-dimethylpropyleneurea (DMPU),
N-methylpyrrolidone (NMP), pyridine or acetonitrile. Preference is
given to tetrahydrofuran.
[0076] Suitable bases for the process step (I-A).fwdarw.(I-B) are
alkali metal hydrides such as potassium hydride or sodium hydride,
alkali metal carbonates such as lithium carbonate, sodium
carbonate, potassium carbonate or caesium carbonate, alkali metal
bicarbonates such as sodium bicarbonate or potassium bicarbonate,
alkali metal alkoxides such as sodium methoxide or potassium
methoxide, sodium ethoxide or potassium ethoxide or potassium
tert-butoxide, amides such as sodium amide, lithium
bis(trimethylsilyl)amide, sodium bis(trimethylsilyl)amide or
potassium bis(trimethylsilyl)amide or lithium diisopropylamide,
organometallic compounds such as butyllithium or phenyllithium, or
organic amines such as triethylamine, diisopropylethylamine,
pyridine, 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) or
1,5-diazabicyclo[4.3.0]non-5-ene (DBN). Preference is given to
lithium bis(trimethylsilyl)amide, sodium bis(trimethylsilyl)amide
and sodium hydride.
[0077] The reaction (I-A).fwdarw.(I-B) is generally carried out in
a temperature range of from -78.degree. C. to +40.degree. C.,
preferably from 0.degree. C. to +20.degree. C. The reaction can be
carried out at atmospheric, elevated or reduced pressure (for
example from 0.5 to 5 bar). The reaction is generally carried out
at atmospheric pressure.
[0078] The preparation processes described can be illustrated in an
exemplary manner by the synthesis schemes below (Schemes 1 to
3):
##STR00012##
##STR00013##
##STR00014##
[0079] The compound of the formula (II) can be prepared by
cyclizing the compound of the formula (X)
##STR00015##
in an inert solvent with hydrazine hydrate to give the compound of
the formula (XI)
##STR00016##
then reacting this compound in an inert solvent in the presence of
a suitable Lewis acid initially with isopentyl nitrite to give the
corresponding diazonium salt and then converting this salt directly
with sodium iodide into the compound of the formula (XII)
##STR00017##
and then reacting this compound in an inert solvent in the presence
of a suitable base with the compound of the formula (XIII)
##STR00018##
[0080] Inert solvents for process step (X).fwdarw.(XI) are alcohols
such as methanol, ethanol, n-propanol, isopropanol, n-butanol,
tert-butanol or 1,2-ethanediol, ethers such as diethyl ether,
dioxane, tetrahydrofuran, glycol dimethyl ether or diethylene
glycol dimethyl ether, hydrocarbons such as benzene, xylene,
toluene, hexane, cyclohexane or mineral oil fractions, or other
solvents such as dimethylformamide (DMF), dimethyl sulphoxide
(DMSO), N,N'-dimethylpropyleneurea (DMPU), N-methylpyrrolidone
(NMP), pyridine, acetonitrile or else water. It is equally possible
to use mixtures of the solvents mentioned. 1,2-Ethanediol is
preferred.
[0081] The reaction (X).fwdarw.(XI) is generally carried out in a
temperature range of from +60.degree. C. to +200.degree. C.,
preferably from +120.degree. C. to +180.degree. C. The reaction can
be carried out at atmospheric, elevated or reduced pressure (for
example from 0.5 to 5 bar). The reaction is generally carried out
at atmospheric pressure.
[0082] Inert solvents for the reaction (XI).fwdarw.(XII) are, for
example, halogenated hydrocarbons such as dichloromethane,
trichloromethane, carbon tetrachloride, trichloroethylene or
chlorobenzene, ethers such as diethyl ether, dioxane,
tetrahydrofuran, glycol dimethyl ether or diethylene glycol
dimethyl ether, or other solvents such as dimethylformamide (DMF),
dimethyl sulphoxide (DMSO), N,N'-dimethylpropyleneurea (DMPU),
N-methylpyrrolidone (NMP), pyridine or acetonitrile. Preference is
given to DMF.
[0083] Suitable Lewis acids for the process step (XI).fwdarw.(XII)
are boron trifluoride/diethyl ether complex, cerium(IV) ammonium
nitrate (CAN), tin(II) chloride, lithium perchlorate, zinc(II)
chloride, indium(III) chloride or indium(III) bromide. Preference
is given to boron trifluoride/diethyl ether complex.
[0084] The reaction (XI).fwdarw.(XII) is generally carried out in a
temperature range of from -78.degree. C. to +40.degree. C.,
preferably from 0.degree. C. to +20.degree. C. The reaction can be
carried out at atmospheric, elevated or reduced pressure (for
example from 0.5 to 5 bar). The reaction is generally carried out
at atmospheric pressure.
[0085] Inert solvents for the reaction (XII)+(XIII).fwdarw.(II)
are, for example, halogenated hydrocarbons such as dichloromethane,
trichloromethane, carbon tetrachloride, trichloroethylene or
chlorobenzene, ethers such as diethyl ether, dioxane,
tetrahydrofuran, glycol dimethyl ether or diethylene glycol
dimethyl ether, or other solvents such as dimethylformamide (DMF),
dimethyl sulphoxide (DMSO), N,N'-dimethylpropyleneurea (DMPU),
N-methylpyrrolidone (NMP), pyridine or acetonitrile. Preference is
given to DMF.
[0086] Suitable bases for the process step (XII)+(XIII).fwdarw.(II)
are alkali metal hydrides such as potassium hydride or sodium
hydride, alkali metal carbonates such as lithium carbonate, sodium
carbonate, potassium carbonate or caesium carbonate, alkali metal
bicarbonates such as sodium bicarbonate or potassium bicarbonate,
alkali metal alkoxides such as sodium methoxide or potassium
methoxide, sodium ethoxide or potassium ethoxide or potassium
tert-butoxide, amides such as sodium amide, lithium
bis(trimethylsilyl)amide, sodium bis(trimethylsilyl)amide or
potassium bis(trimethylsilyl)amide or lithium diisopropylamide,
organometallic compounds such as butyllithium or phenyllithium, or
organic amines such as triethylamine, diisopropylethylamine,
pyridine, 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) or
1,5-diazabicyclo[4.3.0]non-5-ene (DBN). Preference is given to
caesium carbonate.
[0087] The reaction (XII)+(XIII).fwdarw.(II) is generally carried
out in a temperature range of from 0.degree. C. to +60.degree. C.,
preferably from +10.degree. C. to +25.degree. C. The reaction can
be carried out at atmospheric, elevated or reduced pressure (for
example from 0.5 to 5 bar). The reaction is generally carried out
at atmospheric pressure.
[0088] The preparation process described can be illustrated in an
exemplary manner by the synthesis scheme below (Scheme 4):
##STR00019##
[0089] The present invention furthermore provides the compound:
[0090]
5-fluoro-1-(2-fluorobenzyl)-3-iodo-1H-pyrazolo[3,4-b]pyridine
##STR00020##
[0091] The present invention furthermore provides the compound:
[0092] 5-fluoro-3-iodo-1H-pyrazolo[3,4-b]pyridine
##STR00021##
[0093] The compounds of the formulae (III) and (XIII) are
commercially available, known from the literature or can be
prepared in analogy to literature processes.
[0094] The compound of the formula (X) is known from the literature
[cf., for example, Winn M., J. Med. Chem. 1993, 36, 2676-7688; EP
634 413-A1; CN 1613849-A; EP 1626045-A1; WO 2009/018415] and can be
prepared in analogy to literature processses or as shown in the
synthesis scheme below (Scheme 5):
##STR00022##
[0095] The compounds according to the invention act as stimulators
of soluble guanylate cyclase and have an identical or improved
therapeutic profile compared to compounds known from the prior art,
such as, for example, with respect to their in vivo properties such
as, for example, their pharmacokinetic and pharmacodynamic
behaviour and/or their dose-activity relationship and/or their
safety profile. They are therefore suitable for the treatment
and/or prophylaxis of diseases in man and animals.
[0096] The compounds according to the invention lead to
vasorelaxation, to an inhibition of platelet aggregation and to a
reduction in blood pressure, and also to an increase in coronary
blood flow. These effects are mediated via direct stimulation of
soluble guanylate cyclase and intracellular cGMP increase.
Moreover, the compounds according to the invention enhance the
effect of substances increasing the cGMP concentration, such as,
for example, EDRF (endothelium-derived relaxing factor), NO donors,
protoporphyrin IX, arachidonic acid or phenylhydrazine
derivatives.
[0097] The compounds according to the invention are suitable for
the treatment and/or prophylaxis of cardiovascular, pulmonary,
thromboembolic and fibrotic disorders.
[0098] Accordingly, the compounds according to the invention can be
used in medicaments for the treatment and/or prophylaxis of
cardiovascular disorders such as, for example, hypertension, acute
and chronic heart failure, coronary heart disease, stable and
unstable angina pectoris, peripheral and cardiac vascular
disorders, arrhythmias, atrial and ventricular arrhythmias and
impaired conduction such as, for example, atrioventricular blocks
degrees I-III (AB block supraventricular tachyarrhythmia, atrial
fibrillation, atrial flutter, ventricular fibrillation, ventricular
flutter, ventricular tachyarrhythmia, Torsade de pointes
tachycardia, atrial and ventricular extrasystoles, AV-junctional
extrasystoles, Sick-Sinus syndrome, syncopes, AV-nodal re-entry
tachycardia, Wolff-Parkinson-White syndrome, of acute coronary
syndrome (ACS), autoimmune cardiac disorders (pericarditis,
endocarditis, valvolitis, aortitis, cardiomyopathies), shock such
as cardiogenic shock, septic shock and anaphylactic shock,
aneurysms, boxer cardiomyopathy (premature ventricular contraction
(PVC)), for the treatment and/or prophylaxis of thromboembolic
disorders and ischaemias such as myocardial ischaemia, myocardial
infarction, stroke, cardiac hypertrophy, transient and ischaemic
attacks, preeclampsia, inflammatory cardiovascular disorders,
spasms of the coronary arteries and peripheral arteries, oedema
formation such as, for example, pulmonary oedema, cerebral oedema,
renal oedema or oedema caused by heart failure, peripheral
circulatory disturbances, reperfusion damage, arterial and venous
thromboses, microalbuminuria, myocardial insufficiency, endothelial
dsfunction, to prevent restenoses, for example after thrombolysis
therapies, percutaneous transluminal angioplasties (PTA),
transluminal coronary angioplasties (PTCA), heart transplants and
bypass operations, and also micro- and macrovascular damage
(vasculitis), increased levels of fibrinogen and of low-density
lipoprotein (LDL) and increased concentrations of plasminogen
activator inhibitor 1 (PAI-1), and also for the treatment and/or
prophylaxis of erectile dysfunction and female sexual
dysfunction.
[0099] In the context of the present invention, the term heart
failure also includes more specific or related types of disease,
such as acute decompensated heart failure, right heart failure,
left heart failure, global failure, ischaemic cardiomyopathy,
dilated cardiomyopathy, hypertrophic cardiomyopathy, idiopathic
cardiomyopathy, congenital heart defects, heart valve defects,
heart failure associated with heart valve defects, mitral stenosis,
mitral insufficiency, aortic stenosis, aortic insufficiency,
tricuspid stenosis, tricuspid insufficiency, pulmonary valve
stenosis, pulmonary valve insufficiency, combined heart valve
defects, myocardial inflammation (myocarditis), chronic
myocarditis, acute myocarditis, viral myocarditis, diabetic heart
failure, alcoholic cardiomyopathy, cardiac storage disorders, and
diastolic and systolic heart failure.
[0100] In addition, the compounds according to the invention can
also be used for the treatment and/or prophylaxis of
arteriosclerosis, impaired lipid metabolism, hypolipoproteinaemias,
dyslipidaemias, hypertriglyceridaemias, hyperlipidaemias,
hypercholesterolaemias, abetalipoproteinaemias, sitosterolaemia,
xanthomatosis, Tangier disease, adipositas, obesity and of combined
hyperlipidaemias and metabolic syndrome.
[0101] The compounds according to the invention can additionally be
used for the treatment and/or prophylaxis of primary and secondary
Raynaud's phenomenon, of microcirculation impairments,
claudication, peripheral and autonomic neuropathies, diabetic
microangiopathies, diabetic retinopathy, diabetic ulcers on the
extremities, gangrene, CREST syndrome, erythematosis,
onychomycosis, rheumatic disorders and for promoting wound
healing.
[0102] The compounds according to the invention are furthermore
suitable for treating urological disorders such as, for example,
benign prostate syndrome (BPS), benign prostate hyperplasia (BPH),
benign prostate enlargement (BPE), bladder outlet obstruction
(BOO), lower urinary tract syndromes (LUTS, including Feline
Urological Syndrome (FUS)), disorders of the urogenital system
including neurogenic over-active bladder (OAB) and (IC),
incontinence (UI) such as, for example, mixed urinary incontinence,
urge urinary incontinence, stress urinary incontinence or overflow
urinary incontinence (MUI, UUI, SUI, OUI), pelvic pain, benign and
malignant disorders of the organs of the male and femal urogenital
system.
[0103] The compounds according to the invention are furthermore
suitable for the treatment and/or prophylaxis of kidney disorders,
in particular of acute and chronic renal insufficiency and acute
and chronic renal failure. In the context of the present invention,
the term renal insufficiency comprises both acute and chronic
manifestations thereof, as well as underlying or related kidney
diseases such as renal hypoperfusion, intradialytic hypotension,
obstructive uropathy, glomerulopathies, glomerulonephritis, acute
glomerulonephritis, glomerulosclerosis, tubulointerstitial
diseases, nephropathic diseases such as primary and congenital
kidney disease, nephritis, immunological kidney diseases such as
kidney graft rejection and immunocomplex-induced kidney diseases,
nephropathy induced by toxic substances, nephropathy induced by
contrast agents, diabetic and non-diabetic nephropathy,
pyelonephritis, renal cysts, nephrosclerosis, hypertensive
nephrosclerosis and nephrotic syndrome, which can be characterized
diagnostically for example by abnormally reduced creatinine and/or
water excretion, abnormally raised blood concentrations of urea,
nitrogen, potassium and/or creatinine, altered activity of renal
enzymes such as, for example, glutamyl synthetase, altered urine
osmolarity or urine volume, increased microalbuminurea,
macroalbuminurea, laesions on glomerulae and arterioles, tubular
dilatation, hyperphosphataemia and/or need for dialysis. The
present invention also comprises the use of the compounds according
to the invention for the treatment and/or prophylaxis of sequelae
of renal insufficiency, for example pulmonary oedema, heart
failure, uraemia, anaemia, electrolyte disturbances (for example
hypercalaemia, hyponatraemia) and disturbances in bone and
carbohydrate metabolism.
[0104] Furthermore, the compounds according to the invention are
also suitable for the treatment and/or prophylaxis of asthmatic
disorders, pulmonary arterial hypertension (PAH) and other forms of
pulmonary hypertension (PH) including left-heart disease, HIV,
sickle cell anaemia, thromboembolisms (CTEPH), sarkoidosis, COPD or
pulmonary fibrosis-associated pulmonary hypertension,
chronic-obstructive pulmonary disease (COPD), acute respiratory
distress syndrome (ARDS), acute lung injury (ALI),
alpha-1-antitrypsin deficiency (AATD), pulmonary fibrosis,
pulmonary emphysema (for example pulmonary emphysema induced by
cigarette smoke) and cystic fibrosis (CF).
[0105] The compounds described in the present invention also
represent active compounds for controlling central nervous system
diseases characterized by disturbances of the NO/cGMP system. They
are suitable in particular for improving perception, concentration,
learning or memory after cognitive impairments like those occurring
in particular in association with situations/diseases/syndromes
such as mild cognitive impairment, age-associated learning and
memory impairments, age-associated memory losses, vascular
dementia, craniocerebral trauma, stroke, dementia occurring after
strokes (post stroke dementia), post-traumatic craniocerebral
trauma, general concentration impairments, concentration
impairments in children with learning and memory problems,
Alzheimer's disease, Lewy body dementia, dementia with degeneration
of the frontal lobes including Pick's syndrome, Parkinson's
disease, progressive nuclear palsy, dementia with corticobasal
degeneration, amyolateral sclerosis (ALS), Huntington's disease,
demyelination, multiple sclerosis, thalamic degeneration,
Creutzfeld-Jacob dementia, HIV dementia, schizophrenia with
dementia or Korsakoff's psychosis. They are also suitable for the
treatment and/or prophylaxis of central nervous system disorders
such as states of anxiety, tension and depression, CNS-related
sexual dysfunctions and sleep disturbances, and for controlling
pathological disturbances of the intake of food, stimulants and
addictive substances.
[0106] The compounds according to the invention are furthermore
also suitable for controlling cerebral blood flow and thus
represent effective agents for controlling migraine. They are also
suitable for the prophylaxis and control of sequelae of cerebral
infarct (Apoplexia cerebri) such as stroke, cerebral ischaemias and
skull-brain trauma. The compounds according to the invention can
likewise be employed for controlling states of pain and
tinnitus.
[0107] In addition, the compounds according to the invention have
antiinflammatory action and can therefore be used as
antiinflammatory agents for the treatment and/or prophylaxis of
sepsis (SIRS), multiple organ failure (MODS, MOF), inflammatory
disorders of the kidney, chronic intestinal inflammations (IBD,
Crohn's disease, UC), pancreatitis, peritonitis, rheumatoid
disorders, inflammatory skin diseases and inflammatory eye
diseases.
[0108] Furthermore, the compounds according to the invention can
also be used for the treatment and/or prophylaxis of autoimmune
diseases.
[0109] The compounds according to the invention are furthermore
suitable for the treatment and/or prophylaxis of fibrotic disorders
of the internal organs such as, for example, the lung, the heart,
the kidney, the bone marrow and in particular the liver, and also
dermatological fibroses and fibrotic eye disorders. In the context
of the present invention, the term fibrotic disorders includes in
particular the following terms: hepatic fibrosis, cirrhosis of the
liver, pulmonary fibrosis, endomyocardial fibrosis, nephropathy,
glomerulonephritis, interstitial renal fibrosis, fibrotic damage
resulting from diabetes, bone marrow fibrosis and similar fibrotic
disorders, scleroderma, morphea, keloids, hypertrophic scarring
(also following surgical procedures), naevi, diabetic retinopathy,
proliferative vitreoretinopathy and disorders of the connective
tissue (for example sarcoidosis).
[0110] The compounds according to the invention are furthermore
suitable for controlling postoperative scarring, for example as a
result of glaucoma operations.
[0111] The compounds according to the invention can also be used
cosmetically for ageing and keratinized skin.
[0112] Moreover, the compounds according to the invention are
suitable for the treatment and/or prophylaxis of hepatitis,
neoplasms, osteoporosis, glaucoma and gastroparesis.
[0113] The present invention further provides for the use of the
compounds according to the invention for the treatment and/or
prophylaxis of disorders, in particular the disorders mentioned
above.
[0114] The present invention further provides the use of the
compounds according to the invention for the treatment and/or
prophylaxis of heart failure, angina pectoris, hypertension,
pulmonary hypertension, ischaemias, vascular disorders, kidney
failure, thromboembolic disorders, fibrotic disorders and
arteriosclerosis.
[0115] The present invention further provides the compounds
according to the invention for use in a method for the treatment
and/or prophylaxis of heart failure, angina pectoris, hypertension,
pulmonary hypertension, ischaemias, vascular disorders, kidney
failure, thromboembolic disorders, fibrotic disorders and
arteriosclerosis.
[0116] The present invention further provides for the use of the
compounds according to the invention for producing a medicament for
treatment and/or prophylaxis of disorders, in particular the
disorders mentioned above.
[0117] The present invention further provides for the use of the
compounds according to the invention for producing a medicament for
treatment and/or prophylaxis of heart failure, angina pectoris,
hypertension, pulmonary hypertension, ischaemias, vascular
disorders, kidney failure, thromboembolic disorders, fibrotic
disorders and arteriosclerosis.
[0118] The present invention further provides a method for
treatment and/or prophylaxis of disorders, in particular the
disorders mentioned above, using an effective amount of at least
one of the compounds according to the invention.
[0119] The present invention further provides a method for
treatment and/or prophylaxis of heart failure, angina pectoris,
hypertension, pulmonary hypertension, ischaemias, vascular
disorders, kidney failure, thromboembolic disorders, fibrotic
disorders and arteriosclerosis using an effective amount of at
least one of the compounds according to the invention.
[0120] The compounds according to the invention can be employed
alone or, if required, in combination with other active compounds.
The present invention further provides medicaments comprising at
least one of the compounds according to the invention and one or
more further active compounds, especially for the treatment and/or
prophylaxis of the aforementioned disorders. Preferred examples of
suitable active compound combinations include: [0121] organic
nitrates and NO donors, for example sodium nitroprusside,
nitroglycerin, isosorbide mononitrate, isosorbide dinitrate,
molsidomine or SIN-1, and inhaled NO; [0122] compounds which
inhibit the breakdown of cyclic guanosine monophosphate (cGMP),
such as, for example, inhibitors of phosphodiesterases (PDE) 1, 2
and/or 5, in particular PDE 5 inhibitors such as sildenafil,
vardenafil and tadalafil; [0123] agents having an antithrombotic
effect, for example and with preference from the group of platelet
aggregation inhibitors, of anticoagulants or of profibrinolytic
substances; [0124] active compounds which lower blood pressure, for
example and preferably from the group of calcium antagonists,
angiotensin AII antagonists, ACE inhibitors, endothelin
antagonists, renin inhibitors, alpha-receptor blockers,
beta-receptor blockers, mineralocorticoid receptor antagonists, and
of diuretics; and/or [0125] active compounds which alter lipid
metabolism, for example and with preference from the group of
thyroid receptor agonists, cholesterol synthesis inhibitors such
as, by way of example and preferably, HMG-CoA reductase inhibitors
or squalene synthesis inhibitors, of ACAT inhibitors, CETP
inhibitors, MTP inhibitors, PPAR-alpha, PPAR-gamma and/or
PPAR-delta agonists, cholesterol absorption inhibitors, lipase
inhibitors, polymeric bile acid adsorbents, bile acid reabsorption
inhibitors and lipoprotein(a) antagonists;
[0126] Agents having antithrombotic activity preferably mean
compounds from the group of platelet aggregation inhibitors, of
anticoagulants or of profibrinolytic substances.
[0127] In a preferred embodiment of the invention, the compounds
according to the invention are administered in combination with a
platelet aggregation inhibitor such as, by way of example and
preferably, aspirin, clopidogrel, ticlopidin or dipyridamol.
[0128] In a preferred embodiment of the invention, the compounds
according to the invention are administered in combination with a
thrombin inhibitor such as, by way of example and preferably,
ximelagatran, dabigatran, melagatran, bivalirudin or clexane.
[0129] In a preferred embodiment of the invention, the compounds
according to the invention are administered in combination with a
GPIIb/IIIa antagonist such as, by way of example and preferably,
tirofiban or abciximab.
[0130] In a preferred embodiment of the invention, the compounds
according to the invention are administered in combination with a
factor Xa inhibitor such as, by way of example and preferably,
rivaroxaban (BAY 59-7939), DU-176b, apixaban, otamixaban,
fidexaban, razaxaban, fondaparinux, idraparinux, PMD-3112, YM-150,
KFA-1982, EMD-503982, MCM-17, MLN-1021, DX 9065a, DPC 906, JTV 803,
SSR-126512 or SSR-128428.
[0131] In a preferred embodiment of the invention, the compounds
according to the invention are administered in combination with
heparin or a low molecular weight (LMW) heparin derivative.
[0132] In a preferred embodiment of the invention, the compounds
according to the invention are administered in combination with a
vitamin K antagonist such as, by way of example and preferably,
coumarin.
[0133] Agents which lower blood pressure are preferably understood
to mean compounds from the group of calcium antagonists,
angiotensin AII antagonists, ACE inhibitors, endothelin
antagonists, renin inhibitors, alpha-receptor blockers,
beta-receptor blockers, mineralocorticoid receptor antagonists, and
the diuretics.
[0134] In a preferred embodiment of the invention, the compounds
according to the invention are administered in combination with a
calcium antagonist such as, by way of example and preferably,
nifedipine, amlodipine, verapamil or diltiazem.
[0135] In a preferred embodiment of the invention, the compounds
according to the invention are administered in combination with an
alpha-1 receptor blocker such as, by way of example and preferably,
prazosin.
[0136] In a preferred embodiment of the invention, the compounds
according to the invention are administered in combination with a
beta receptor blocker such as, by way of example and preferably,
propranolol, atenolol, timolol, pindolol, alprenolol, oxprenolol,
penbutolol, bupranolol, metipranolol, nadolol, mepindolol,
carazalol, sotalol, metoprolol, betaxolol, celiprolol, bisoprolol,
carteolol, esmolol, labetalol, carvedilol, adaprolol, landiolol,
nebivolol, epanolol or bucindolol.
[0137] In a preferred embodiment of the invention, the compounds
according to the invention are administered in combination with an
angiotensin AII antagonist such as, by way of example and
preferably, losartan, candesartan, valsartan, telmisartan or
embusartan.
[0138] In a preferred embodiment of the invention, the compounds
according to the invention are administered in combination with an
ACE inhibitor such as, by way of example and preferably, enalapril,
captopril, lisinopril, ramipril, delapril, fosinopril, quinopril,
perindopril or trandopril.
[0139] In a preferred embodiment of the invention, the compounds
according to the invention are administered in combination with an
endothelin antagonist such as, by way of example and preferably,
bosentan, darusentan, ambrisentan or sitaxsentan.
[0140] In a preferred embodiment of the invention, the compounds
according to the invention are administered in combination with a
renin inhibitor such as, for example and preferably, aliskiren,
SPP-600 or SPP-800.
[0141] In a preferred embodiment of the invention, the compounds
according to the invention are administered in combination with a
mineralocorticoid receptor antagonist such as, for example and
preferably, spironolactone or eplerenone.
[0142] In a preferred embodiment of the invention, the compounds
according to the invention are administered in combination with a
loop diuretic such as, for example, furosemide, torasemide,
bumetanide and piretanide, with potassium-sparing diuretics such
as, for example, amiloride and triamterene, with aldosterone
antagonists such as, for example, spironolactone, potassium
canrenoate and eplerenone and also thiazide diuretics such as, for
example, hydrochlorothiazide, chlorthalidone, xipamide and
indapamide.
[0143] Active compounds which alter lipid metabolism are preferably
understood to mean compounds from the group of CETP inhibitors,
thyroid receptor agonists, cholesterol synthesis inhibitors such as
HMG-CoA reductase inhibitors or squalene synthesis inhibitors, of
ACAT inhibitors, MTP inhibitors, PPAR-alpha, PPAR-gamma and/or
PPAR-delta agonists, cholesterol absorption inhibitors, polymeric
bile acid adsorbents, bile acid reabsorption inhibitors, lipase
inhibitors and lipoprotein(a) antagonists;
[0144] In a preferred embodiment of the invention, the compounds
according to the invention are administered in combination with a
CETP inhibitor such as, by way of example and preferably,
dalcetrapib, BAY 60-5521, anacetrapib or CETP vaccine (CETi-1).
[0145] In a preferred embodiment of the invention, the compounds
according to the invention are administered in combination with a
thyroid receptor agonist such as, by way of example and preferably,
D-thyroxin, 3,5,3'-triiodothyronin (T3), CGS 23425 or axitirome
(CGS 26214).
[0146] In a preferred embodiment of the invention, the compounds
according to the invention are administered in combination with a
HMG-CoA reductase inhibitor from the class of the statins such as,
by way of example and preferably, lovastatin, simvastatin,
pravastatin, fluvastatin, atorvastatin, rosuvastatin or
pitavastatin.
[0147] In a preferred embodiment of the invention, the compounds
according to the invention are administered in combination with a
squalene synthesis inhibitor such as, by way of example and
preferably, BMS-188494 or TAK-475.
[0148] In a preferred embodiment of the invention, the compounds
according to the invention are administered in combination with an
ACAT inhibitor such as, by way of example and preferably,
avasimibe, melinamide, pactimibe, eflucimibe or SMP-797.
[0149] In a preferred embodiment of the invention, the compounds
according to the invention are administered in combination with an
MTP inhibitor such as, by way of example and preferably,
implitapide, BMS-201038, R-103757 or JTT-130.
[0150] In a preferred embodiment of the invention, the compounds
according to the invention are administered in combination with a
PPAR-gamma agonist such as, by way of example and preferably,
pioglitazone or rosiglitazone.
[0151] In a preferred embodiment of the invention, the compounds
according to the invention are administered in combination with a
PPAR-delta agonist such as, for example and preferably, GW 501516
or BAY 68-5042.
[0152] In a preferred embodiment of the invention, the compounds
according to the invention are administered in combination with a
cholesterol absorption inhibitor such as, by way of example and
preferably, ezetimibe, tiqueside or pamaqueside.
[0153] In a preferred embodiment of the invention, the compounds
according to the invention are administered in combination with a
lipase inhibitor, a preferred example being orlistat.
[0154] In a preferred embodiment of the invention, the compounds
according to the invention are administered in combination with a
polymeric bile acid adsorbent such as, by way of example and
preferably, cholestyramine, colestipol, colesolvam, CholestaGel or
colestimide.
[0155] In a preferred embodiment of the invention, the compounds
according to the invention are administered in combination with a
bile acid reabsorption inhibitor such as, by way of example and
preferably, ASBT (=IBAT) inhibitors, for example AZD-7806, S-8921,
AK-105, BARI-1741, SC-435 or SC-635
[0156] In a preferred embodiment of the invention, the compounds
according to the invention are administered in combination with an
lipoprotein(a) antagonist such as, by way of example and
preferably, gemcabene calcium (CI-1027) or nicotinic acid.
[0157] The present invention further provides medicaments which
comprise at least one compound according to the invention,
typically together with one or more inert, nontoxic,
pharmaceutically suitable auxiliaries, and the use thereof for the
aforementioned purposes.
[0158] The compounds according to the invention may act
systemically and/or locally. For this purpose, they can be
administered in a suitable manner, for example by the oral,
parenteral, pulmonal, nasal, sublingual, lingual, buccal, rectal,
dermal, transdermal, conjunctival, otic route, or as an implant or
stent.
[0159] The compounds according to the invention can be administered
in administration forms suitable for these administration
routes.
[0160] Suitable administration forms for oral administration are
those which work according to the prior art, which release the
compounds according to the invention rapidly and/or in a modified
manner and which contain the compounds according to the invention
in crystalline and/or amorphized and/or dissolved form, for example
tablets (uncoated or coated tablets, for example with gastric
juice-resistant or retarded-dissolution or insoluble coatings which
control the release of the compound according to the invention),
tablets or films/wafers which disintegrate rapidly in the oral
cavity, films/lyophilizates or capsules (for example hard or soft
gelatin capsules), sugar-coated tablets, granules, pellets,
powders, emulsions, suspensions, aerosols or solutions.
[0161] Parenteral administration can bypass an absorption step
(e.g. intravenously, intraarterially, intracardially, intraspinally
or intralumbally) or include an absorption (e.g. intramuscularly,
subcutaneously, intracutaneously, percutaneously or
intraperitoneally). Administration forms suitable for parenteral
administration include preparations for injection and infusion in
the form of solutions, suspensions, emulsions, lyophilizates or
sterile powders.
[0162] For the other administration routes, suitable examples are
inhalable medicament forms (including powder inhalers, nebulizers),
nasal drops, solutions or sprays, tablets, films/wafers or capsules
for lingual, sublingual or buccal administration, suppositories,
car or eye preparations, vaginal capsules, aqueous suspensions
(lotions, shaking mixtures), lipophilic suspensions, ointments,
creams, transdermal therapeutic systems (e.g. patches), milk,
pastes, foams, sprinkling powders, implants or stents.
[0163] Oral or parenteral administration is preferred, especially
oral administration.
[0164] The compounds according to the invention can be converted to
the administration forms mentioned. This can be done in a manner
known per se, by mixing with inert, nontoxic, pharmaceutically
suitable excipients. These excipients include carriers (for example
microcrystalline cellulose, lactose, mannitol), solvents (e.g.
liquid polyethylene glycols), emulsifiers and dispersing or wetting
agents (for example sodium dodecylsulphate, polyoxysorbitan
oleate), binders (for example polyvinylpyrrolidone), synthetic and
natural polymers (for example albumin), stabilizers (e.g.
antioxidants, for example ascorbic acid), dyes (e.g. inorganic
pigments, for example iron oxides) and flavour and/or odour
correctants.
[0165] In general, it has been found to be advantageous in the case
of parenteral administration to administer amounts of about 0.001
to 1 mg/kg, preferably about 0.01 to 0.5 mg/kg, of body weight to
achieve effective results. In the case of oral administration, the
dosage is about 0.01 to 100 mg/kg, preferably about 0.01 to 20
mg/kg and most preferably 0.1 to 10 mg/kg of body weight.
[0166] It may nevertheless be necessary where appropriate to
deviate from the stated amounts, specifically as a function of the
body weight, route of administration, individual response to the
active compound, nature of the preparation and time or interval
over which administration takes place. For instance, in some cases,
less than the aforementioned minimum amount may be sufficient,
while in other cases the upper limit mentioned must be exceeded. In
the case of administration of relatively large amounts, it may be
advisable to divide these into several individual doses over the
course of the day.
[0167] The working examples which follow illustrate the invention.
The invention is not limited to the examples.
[0168] The percentages in the tests and examples which follow are,
unless indicated otherwise, percentages by weight; parts are parts
by weight. Solvent ratios, dilution ratios and concentration data
for liquid/liquid solutions are based in each case on volume.
A. EXAMPLES
Abbreviations and Acronyms
[0169] aq. aqueous solution [0170] calc. calculated [0171] br s
broad singlet (in NMR) [0172] DCI direct chemical ionization (in
MS) [0173] dec. decomposition point [0174] DMF dimethylformamide
[0175] DMSO dimethyl sulphoxide [0176] DSC dynamic differential
calorimetry [0177] eq. equivalent(s) [0178] ESI electrospray
ionization (in MS) [0179] Et ethyl [0180] fnd. found [0181] h
hour(s) [0182] HPLC high-pressure high-performance liquid
chromatography [0183] HRMS high-resolution mass spectrometry [0184]
conc. concentrated [0185] LC-MS liquid chromatography-coupled mass
spectrometry [0186] LiHMDS lithium hexamethyldisilazide [0187] Me
methyl [0188] min minute(s) [0189] MS mass spectrometry [0190] NMR
nuclear magnetic resonance spectrometry [0191] Pd.sub.2dba.sub.3
tris(dibenzylideneacetone)dipalladium [0192] Ph phenyl [0193] PLM
polarized light microscope [0194] RT room temperature [0195]
R.sub.t retention time (in HPLC) [0196] TGA thermogravimetric
analysis [0197] THF tetrahydrofuran [0198] UV ultraviolet
spectrometry [0199] v/v volume to volume ratio (of a solution)
LC/MS Methods:
[0200] Method 1:
[0201] MS instrument type: Waters ZQ; apparatus type HPLC: Agilent
1100 Series; UV DAD; column: Thermo Hypersil GOLD 3 it 20
mm.times.4 mm; mobile phase A: 1 l of water+0.5 ml of 50% strength
formic acid, mobile phase B: 1 l of acetonitrile+0.5 ml of 50%
strength formic acid; gradient: 0.0 min 100% A.fwdarw.3.0 min 10%
A.fwdarw.4.0 min 10% A.fwdarw.4.1 min 100% A (flow rate 2.5
ml/min); oven: 55.degree. C.; flow rate: 2 ml/min; UV detection:
210 nm.
[0202] Method 2:
[0203] Instrument: Waters ACQUITY SQD UPLC System; column: Waters
Acquity UPLC HSS T3 1.8.mu. 50.times.1 mm; mobile phase A: 1 l of
water+0.25 ml of 99% strength formic acid, mobile phase B: 1 l of
acetonitrile+0.25 ml of 99% strength formic acid; gradient: 0.0 min
90% A.fwdarw.1.2 min 5% A.fwdarw.2.0 min 5% A oven: 50.degree. C.;
flow rate: 0.40 ml/min; UV detection: 210-400 nm.
[0204] Method 3:
[0205] Instrument: Micromass Quattro Premier with Waters UPLC
Acquity; column: Thermo Hypersil GOLD 1.9.mu. 50 mm.times.1 mm;
mobile phase A: 1 l of water+0.5 ml of 50% strength formic acid,
mobile phase B: 1 l of acetonitrile+0.5 ml of 50% strength formic
acid; gradient: 0.0 min 90% A 0.1 min 90% A 1.5 min 10% A 2.2 min
10% A; oven 50.degree. C.; flow rate: 0.33 ml/min; UV detection:
210 nm.
General Methods:
[0206] PLM:
[0207] The polarized light microscopy was carried out using a
Clemex PS3 polarized light microscope particle size system with a
Leica DM microscope procided with 50.times., 100.times.,
200.times., and 500.times. lenses, a high-resolution monochrome
1600.times.1200 pixel digital camera and a motorized X-Y Marzhauser
station (controlled by a Clemex ST-2000 controller). The samples of
the crystalline material were measured on a glass slide
(76.times.26 mm) in a drop of oil, the sample being covered with a
cover glass (22.times.40 mm).
[0208] DSC:
[0209] The melting points were determined by dynamic differential
calorimetry. The determination was carried out using a
Mettler-Toledo 823.sup.e DSC instrument provided with a TSO801RO
sample robot and STAR.sup.e software. About 1.5 to 3 mg of the
sample were weighed out into a small aluminium pan, which was then
closed with a perforated cap. The heat flow was measured in a
temperature range of from 30 to 400.degree. C. at a heating rate of
10.degree. C./min and under an argon stream of 30 ml/min.
[0210] TGA:
[0211] The thermogravimetric analysis was carried out using a
Mettler-Toledo TGA/SDTA851.sup.e TGA instrument provided with a
TSO801RO sample robot and STAR.sup.e software. About 1.5 to 3 mg of
the sample were weighed out into a small open aluminium pan (100
.mu.l). The sample weight was measured in a temperature range of
from 30 to 400.degree. C. at a heating rate of 10.degree. C./min
and under an argon stream of 30 ml/min.
[0212] The elemental analyses were carried out by Currenta GmbH
& Co. using methods known to the person skilled in the art, in
accordance with industry norm DIN-ISO 17025.
Starting Materials and Intermediates:
Example 1A
2,6-Dichloro-5-fluoronicotinamide
##STR00023##
[0214] A suspension of 25 g (130.90 mmol) of
2,6-dichloro-5-fluoro-3-cyanopyridine in conc. sulphuric acid (125
ml) was stirred at 60-65.degree. C. for 1 h. After cooling to RT,
the contents of the flask were poured into ice-water and extracted
three times with ethyl acetate (100 ml each time). The combined
organic phases were washed with water (100 ml) and then with
saturated aqueous sodium hydrogen carbonate solution (100 ml),
dried and concentrated on a rotary evaporator. The material
obtained was dried under high vacuum.
[0215] Yield: 24.5 g (90% of theory)
[0216] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta.=7.95 (br s,
1H), 8.11 (br s, 1H), 8.24 (d, 1H).
Example 2A
2-Chloro-5-fluoronicotinamide
##STR00024##
[0218] At RT, 44 g (210.58 mmol) of
2,6-dichloro-5-fluoronicotinamide were added to a suspension of
21.9 g (335.35 mmol) of zinc in methanol (207 ml). Acetic acid
(18.5 ml) was then added, and the mixture was heated with stirring
at reflux for 24 h. The contents of the flask were then decanted
from the zinc, and ethyl acetate (414 ml) and saturated aqueous
sodium hydrogen carbonate solution (414 ml) were added, followed by
intense extractive stirring. Subsequently the reaction mixture was
filtered with suction through kieselguhr and the filter product was
washed three times with ethyl acetate (517 ml each time). The
organic phase was separated off and the aqueous phase was washed
with ethyl acetate (258 ml). The combined organic phases were
washed once with saturated aqueous sodium hydrogen carbonate
solution (414 ml), dried and concentrated under reduced pressure.
Dichloromethane (388 ml) was added to the crystals obtained in this
manner, and the mixture was stirred for 20 min. The mixture was
once more filtered off with suction, washed with diethyl ether and
sucked dry.
[0219] Yield: 20.2 g (53% of theory)
[0220] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta.=7.87 (br s,
1H), 7.99 (dd, 1H), 8.10 (br s, 1H), 8.52 (d, 1H).
Example 3A
2-Chloro-5-fluoronicotinonitrile
##STR00025##
[0222] 81.2 ml (582.25 mmol) of triethylamine were added to a
suspension of 46.2 g (264.66 mmol) of 2-chloro-5-fluoronicotinamide
in dichloromethane (783 ml), and the mixture was cooled to
0.degree. C. Then, with stirring, 41.12 ml (291.13 mmol) of
trifluoroacetic anhydride were added slowly dropwise and the
mixture was stirred at 0.degree. C. for 1.5 h. The reaction
solution was subsequently washed twice with saturated aqueous
sodium bicarbonate solution (391 ml each time), dried and
concentrated under reduced pressure.
[0223] Yield: 42.1 g (90% of theory)
[0224] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta.=8.66 (dd, 1H),
8.82 (d, 1H).
Example 4A
5-Fluoro-1H-pyrazolo[3,4-b]pyridine-3-amine
##STR00026##
[0226] A suspension of 38.5 g (245.93 mmol) of
2-chloro-5-fluoronicotinonitrile was initially charged in
1,2-ethanediol (380 ml), and hydrazine hydrate (119.6 ml, 2.459
mol) was then added. The mixture was heated under reflux with
stirring for 4 h. The product precipitated on cooling. Water (380
ml) was added to the yellow crystals, and the mixture was subjected
to extractive stirring at RT for 10 min. The suspension was then
filtered with suction over a frit, and the filter product was
washed with water (200 ml) and with -10.degree. C. cold THF (200
ml). The residue was dried under high vacuum over phosphorus
pentoxide.
[0227] Yield: 22.8 g (61% of theory)
[0228] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta.=5.54 (s, 2H),
7.96 (dd, 1H), 8.38 (m, 1H), 12.07 (m, 1H).
Example 5A
5-Fluoro-3-iodo-1H-pyrazolo[3,4-b]pyridine
##STR00027##
[0230] 10 g (65.75 mmol) of
5-fluoro-1H-pyrazolo[3,4-b]pyridine-3-amine were initially charged
in THF (329 ml), and the mixture was cooled to 0.degree. C. 16.65
ml (131.46 mmol) of boron trifluoride diethyl ether complex were
then added slowly. The reaction mixture was cooled further to
-10.degree. C. A solution of 10.01 g (85.45 mmol) of isopentyl
nitrite in THF (24.39 ml) was then added slowly, and the mixture
was stirred for a further 30 min. The mixture was diluted with cold
diethyl ether (329 ml) and the resulting solid was isolated by
filtration. A little at a time, the diazonium salt thus prepared
was added to a cold (0.degree. C.) solution of 12.81 g (85.45 mmol)
of sodium iodide in acetone (329 ml), and the mixture was stirred
at RT for 30 min. The reaction mixture was poured into ice-water
(1.8 l) and extracted twice with ethyl acetate (487 ml each time).
The collected organic phases were washed with saturated aqueous
sodium chloride solution (244 ml), dried, filtered and
concentrated. This gave 12.1 g (86% purity, 60% of theory) of the
desired compound in the form of a brown solid. The crude product
was reacted without further purification.
[0231] LC-MS (method 1): R.sub.t=1.68 min; MS (ESIpos): m/z=264
(M+H).sup.+.
Example 6A
5-Fluoro-1-(2-fluorobenzyl)-3-iodo-1H-pyrazolo[3,4-b]pyridine
##STR00028##
[0233] 141 g (462.11 mmol) of the compound from Example 5A were
introduced into DMF (2538 ml), and 96.09 g (508.32 mmol) of
2-fluorobenzyl bromide and 165.62 g (508.32 mmol) of caesium
carbonate were then added. The mixture was stirred at RT for two
hours. The reaction mixture was then poured into saturated aqueous
sodium chloride solution (13 670 ml) and extracted twice with ethyl
acetate (5858 ml). The collected organic phases were washed with
saturated aqueous sodium chloride solution (3905 ml), dried,
filtered and concentrated. The residue was chromatographed on
silica gel (mobile phase: petroleum ether/ethyl acetate 97:3) and
the product fractions were concentrated. The resulting solid was
dissolved in dichloromethane and washed once with saturated aqueous
sodium thiosulphate solution (500 ml) and then with saturated
aqueous sodium chloride solution (500 ml). The product was
concentrated to dryness and the residue was suspended in diethyl
ether, isolated by filtration with suction and dried under high
vacuum. This gave 106.6 g (62% of theory) of the desired
compound.
[0234] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta.=5.73 (s, 2H),
7.13-7.26 (m, 3H), 7.33-7.41 (m, 1H), 7.94 (dd, 1H), 8.69-8.73 (m,
1H).
Example 7A
2-[5-Fluoro-1-(2-fluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl]-5-nitropyri-
midine-4,6-diamine
##STR00029##
[0236] Under argon, 860 mg (2.32 mmol) of the compound from Example
6A were introduced into 1,4-dioxane (86 ml), and the reaction
mixture was flushed with argon for 10 min. Then 3.51 ml (6.95 mmol)
of hexabutylditin and 483 mg (2.55 mmol) of
2-chloro-5-nitropyrimidine-4,6-diamine (prepared by the method of
Helvetica Chimica Acta (1951), 34, 835-40) were added. Subsequently
860 mg (0.744 mmol) of tetrakis(triphenylphosphine)palladium(0)
were added and the reaction mixture was heated at reflux overnight.
The mixture was then cooled to RT, water was added and the mixture
was extracted twice with ethyl acetate. The collected organic
phases were dried over sodium sulphate, filtered and concentrated.
The residue was subjected to extractive stirring in ethyl acetate,
and the solid was isolated by filtration and dried under high
vacuum. This gave 355 mg (62% purity, 24% of theory) of the desired
compound. The crude product was reacted without further
purification.
[0237] LC-MS (method 2): R.sub.t=1.03 min
[0238] MS (ESIpos): m/z=399 (M-H).sup.+
Example 8A
5-Fluoro-1-(2-fluorobenzyl)-1H-pyrazolo[3,4-b]pyridine-3-carbonitrile
##STR00030##
[0240] A suspension of 16.03 g (43.19 mmol) of
5-fluoro-1-(2-fluorobenzyl)-3-iodo-1H-pyrazolo[3,4-b]pyridine
(Example 6A) and 4.25 g (47.51 mmol) of copper cyanide was
initially charged in DMSO (120 ml) and stirred at 150.degree. C.
for 2 h. After cooling, the contents of the flask were cooled to
about 40.degree. C. and poured into a solution of conc. aqueous
ammonia (90 ml) and water (500 ml), ethyl acetate (200 ml) was
added and the mixture was subjected to brief extractive stirring.
The aqueous phase was separated off and extracted two more times
with ethyl acetate (200 ml each time). The combined organic phases
were washed twice with 10% strength aqueous sodium chloride
solution (100 ml each time), dried and concentrated under reduced
pressure. The crude product was reacted without further
purification.
[0241] Yield: 11.1 g (91% of theory)
[0242] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta.=5.87 (s, 2H),
7.17-7.42 (m, 4H), 8.52 (dd, 1H), 8.87 (dd, 1H).
Example 9A
5-Fluoro-1-(2-fluorobenzyl)-1H-pyrazolo[3,4-b]pyridine-3-carboximidamide
acetate
##STR00031##
[0244] 11.1 g (41.07 mmol) of
5-fluoro-1-(2-fluorobenzyl)-1H-pyrazolo[3,4-b]pyridine-3-carbonitrile
(Example 8A) were added to 2.22 g (41.07 mmol) of sodium methoxide
in methanol (270 ml), and the mixture was stirred at RT for 2 h.
2.64 g (49.29 mmol) of ammonium chloride and acetic acid (9.17 ml)
were then added, and the mixture was heated at reflux overnight.
The mixture was then concentrated to dryness and the residue was
taken up in water (100 ml) and ethyl acetate (100 ml) and adjusted
to a pH of 10 using 2N aqueous sodium hydroxide solution. The
mixture was stirred intensively at RT for about 1 h. The resulting
suspension was filtered with suction and the filter product was
washed with ethyl acetate (100 ml), with water (100 ml) and once
more with ethyl acetate (100 ml). The residue was dried under high
vacuum over phosphorus pentoxide.
[0245] Yield: 9.6 g (78% of theory)
[0246] MS (ESIpos): m/z=288 (M+H).sup.+
[0247] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta.=1.85 (s, 3H),
5.80 (s, 2H), 7.14-7.25 (m, 3H), 7.36 (m, 1H), 8.42 (dd, 1H), 8.72
(dd, 1H).
Example 10A
2-[5-Fluoro-1-(2-fluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl]-5-[(E)-phen-
yldiazenyl]pyrimidine-4,6-diamine
##STR00032##
[0249] With stirring, 3.85 g (41.34 mmol) of aniline were added to
water (40 ml) and conc. hydrochloric acid (7.07 ml), and this
mixture was cooled to 0.degree. C. A solution of 2.85 g (41.34
mmol) of sodium nitrite in water (21 ml) was then added dropwise at
between 0.degree. C. and 5.degree. C., followed by stirring at
0.degree. C. for 15 min. Thereafter, at 0.degree. C., a solution of
4.28 g (52.25 mmol) of sodium acetate in water (19 ml) was added
rapidly dropwise, and then, with thorough stirring, a solution of
2.73 g (41.34 mmol) of malononitrile in ethanol (10 ml) was added
dropwise. After 2 h at 0.degree. C., the resulting precipitate was
isolated by filtration with suction and washed three times with
water (50 ml each time) and with petroleum ether (50 ml). The
residue, still moist, was dissolved in DMF (46 ml) and added
dropwise at precisely 85.degree. C. to a solution of 9.5 g (33.07
mmol) of
5-fluoro-1-(2-fluorobenzyl)-1H-pyrazolo[3,4-b]pyridine-3-carboximidami-
de acetate (Example 9A) in DMF (46 ml) and triethylamine (5.76 ml).
The mixture was then stirred at 100.degree. C. for 4 h and left to
cool to RT overnight. The mixture was poured into water (480 ml)
and subjected to extractive stirring at RT for 1 h. After the
precipitate had been isolated by filtration with suction, it was
washed twice with water (100 ml each time) and twice with methanol
(50 ml each time) and then dried under a high vacuum.
[0250] Yield: 9.6 g (59% of theory)
[0251] LC-MS (method 2): R.sub.t=1.21 min
[0252] MS (ESIpos): m/z=458 (M+H).sup.+
Example 11A
2-[5-Fluoro-1-(2-fluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl]pyrimidine-4-
,5,6-triamine
##STR00033##
[0254] Variant A:
[0255] Preparation Starting from Example 7A:
[0256] In pyridine (30 ml), 378 mg (0.949 mmol) of the compound
from Example 7A were introduced and then 143 mg (0.135 mmol) of
palladium (10% on carbon) were added. The mixture was hydrogenated
overnight at RT under standard hydrogen pressure. The suspension
was then filtered through kieselguhr and the filtercake was washed
with ethanol. The filtrate was concentrated and yielded 233 mg (81%
purity, 51% of theory) of the desired compound, which was reacted
without further purification.
[0257] Variant B:
[0258] Preparation Starting from Example 10A:
[0259] In DMF (800 ml), 39.23 g (85.75 mmol) of the compound from
Example 10A were introduced and then 4 g of palladium (10% on
carbon) were added. The mixture was hydrogenated with stirring
overnight under standard hydrogen pressure. The batch was filtered
over kieselguhr and the filter product was washed with a little DMF
and then with a little methanol, and concentrated to dryness. The
residue was admixed with ethyl acetate and stirred vigorously, and
the precipitate was filtered off with suction, washed with ethyl
acetate and diisopropyl ether and dried under a high vacuum over
Sicapent.
[0260] Yield: 31.7 g (100% of theory)
[0261] LC-MS (method 2): R.sub.t=0.78 min
[0262] MS (ESIpos): m/z=369 (M+H).sup.+
WORKING EXAMPLES
Example 1
Methyl
{4,6-diamino-2-[5-fluoro-1-(2-fluorobenzyl)-1H-pyrazolo[3,4-b]pyrid-
in-3-yl]pyrimidin-5-yl}carbamate
##STR00034##
[0264] In pyridine (600 ml), 31.75 g (86.20 mmol) of the compound
from Example 11A were introduced under argon and cooled to
0.degree. C. Then a solution of 6.66 ml (86.20 mmol) of methyl
chloroformate in dichloromethane (10 ml) was added dropwise and the
mixture was stirred at 0.degree. C. for 1 h. Thereafter the
reaction mixture was brought to RT, concentrated under reduced
pressure and co-distilled repeatedly with toluene. The residue was
stirred with water/ethanol and then filtered off on a frit, after
which it was washed with ethanol and ethyl acetate. Subsequently
the residue was again stirred with diethyl ether, isolated by
filtration with suction and then dried under a high vacuum.
[0265] Yield: 24.24 g (65% of theory)
[0266] LC-MS (method 2): R.sub.t=0.79 min
[0267] MS (ESIpos): m/z=427 (M+H).sup.+
[0268] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta.=3.62 (br. s,
3H), 5.79 (s, 2H), 6.22 (br. s, 4H), 7.10-7.19 (m, 2H), 7.19-7.26
(m, 1H), 7.32-7.40 (m, 1H), 7.67 and 7.99 (2 br. s, 1H), 8.66 (m,
1H), 8.89 (dd, 1H).
Example 2
Methyl
{4,6-diamino-2-[5-fluoro-1-(2-fluorobenzyl)-1H-pyrazolo[3,4-b]pyrid-
in-3-yl]pyrimidin-5-yl}methylcarbamate
##STR00035##
[0270] A quantity of 200 mg (0.469 mmol) of methyl
4,6-diamino-2-[5-fluoro-1-(2-fluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl-
]pyrimidin-5-ylcarbamate (Example 1) was introduced in THF (5 ml)
at 0.degree. C. Then 0.704 ml (0.704 mmol) of lithium
hexamethyldisilazane solution (1M in THF) was added and the mixture
was stirred at this temperature for 20 min. Subsequently 43.8
(0.704 mmol) of iodomethane were added and the mixture was warmed
to RT. After 1 h at this temperature, reaction was terminated with
water (1 ml) and the reaction mixture was concentrated, the residue
being separated by means of preparative RP-HPLC (water (+0.05%
formic acid)-acetonitrile gradient).
[0271] Yield: 90 mg (44% of theory)
[0272] LC-MS (method 2): R.sub.t=0.85 min
[0273] MS (ESIpos): m/z=441 (M+H).sup.+
[0274] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta.=3.00 (s, 3H),
3.53 and 3.66 (2s, 3H), 5.81 (s, 2H), 6.57 (br. s, 4H), 7.13 (m,
2H), 7.22 (m, 1H), 7.35 (m, 1H), 8.67 (m, 1H), 8.87 (dd, 1H).
Example 3
[0275] Methyl
{4,6-diamino-2-[5-fluoro-1-(2-fluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-y-
l]pyrimidin-5-yl}(2,2,2-trifluoroethyl)carbamate
##STR00036##
[0276] A quantity of 3.470 g (8.138 mmol) of the compound from
Example 1 was suspended in 35 ml of THF, admixed at 0.degree. C.
with 358 mg (8.952 mmol) of sodium hydride (60% suspension in
mineral oil) and stirred at 0.degree. C. for 90 min, in the course
of which a solution was formed. A quantity of 2.519 g (8.952 mmol)
of 2,2,2-trifluoroethyl trichloromethanesulphonate was added and
the mixture was stirred at RT for 48 h. It was then stirred with
water and concentrated on a rotary evaporator. The residue was
taken up in ethyl acetate, and the organic phase was washed twice
with water and dried over sodium sulphate. This gave 5.005 g of the
target compound (79% of theory, purity by HPLC 65%). A quantity of
250 mg of the residue was purified by means of preparative HPLC
(mobile phase: methanol/water, gradient 30:70.fwdarw.90:10).
[0277] LC-MS (method 2): R.sub.t=0.97 min; MS (EIpos): m/z=509
(M+H).sup.+.
[0278] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm]=3.63 (s,
3H), 4.06-4.15 (m, 2H), 5.80 (s, 2H), 6.46 (br s, 4H) 7.11-7.15 (m,
2H), 7.20-7.25 (m, 1H), 7.33-7.38 (m, 1H), 8.66 (dd, 1H), 8.91 (dd,
1H).
Example 4
Methyl
{4,6-diamino-2-[5-fluoro-1-(2-fluorobenzyl)-1H-pyrazolo[3,4-b]pyrid-
in-3-yl]pyrimidin-5-yl}carbamate hydrochloride
##STR00037##
[0280] A solution of 100 mg (0.235 mmol) of Example 1 in 2 ml of
1,4-dioxane was prepared in a brown 5 ml glass bottle. In
succession, 2 ml of isopropanol and 235 .mu.l (0.235 mmol) of 1M
hydrochloric acid were added to this solution, and the solution was
stirred at RT until the solvents had evaporated. Air-drying gave
102 mg (94% of theory) of the title compound.
[0281] PLM (100.times.): crystalline
[0282] DSC: 224.degree. C. (dec., .DELTA.H=189 J/g)
[0283] TGA: 1% weight loss at 80.degree. C.
[0284] LC-MS (method 3): R.sub.t=0.91 min
[0285] MS (ESIpos): m/z=427 (M+H).sup.+
[0286] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm]=3.35 and
3.65 (2 s, 3H), 5.92 (s, 2H), 7.15 (dd, 1H), 7.25 (m, 2H), 7.37 (m,
1H), 7.75 (br s, 4H), 8.08 and 8.39 (2 s, 1H), 8.82 (m, 2H), 13.2
(br s, 1H).
[0287] Elemental analysis for
C.sub.19H.sub.16F.sub.2N.sub.8O.sub.2+HCl:
[0288] calculated: % C, 49.31; % H, 3.70; % N, 24.21;
[0289] measured: % C, 49.5; % H, 3.7; % N, 24.3.
Example 5
Methyl
{4,6-diamino-2-[5-fluoro-1-(2-fluorobenzyl)-1H-pyrazolo[3,4-b]pyrid-
in-3-yl]pyrimidin-5-yl}carbamate sulphate
##STR00038##
[0291] A solution of 100 mg (0.235 mmol) of Example 1 in 2 ml of
1,4-dioxane was prepared in a brown 5 ml glass bottle. In
succession, 2 ml of isopropanol and a solution of 938 .mu.l (0.235
mmol) of 0.25M sulphuric acid were added to this solution, and the
solution was stirred at RT until the solvents had evaporated.
Air-drying gave 103 mg (83.7% of theory) of the title compound.
[0292] PLM (100.times.): crystalline
[0293] DSC: 242.degree. C. (dec., .DELTA.H=115 J/g)
[0294] TGA: no weight loss prior to decomposition
[0295] LC-MS (method 3): R.sub.t=0.91 min
[0296] MS (ESIpos): m/z=427 (M+H).sup.+
[0297] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm]=3.56 and
3.66 (2 s, 3H), 5.93 (s, 2H), 7.16 (m, 2H), 7.25 (dd, 1H), 7.38 (m,
1H), 7.59 (br s, 4H), 8.03 and 8.32 (2 s, 1H), 8.82 (m, 2H), 13.0
(br s, 1H).
[0298] Elemental analysis for
C.sub.19H.sub.16F.sub.2N.sub.8O.sub.2H.sub.2SO.sub.4:
[0299] calculated: % C, 43.51; % H, 3.46; % N, 21.37;
[0300] measured: % C, 43.6; % H, 3.4; % N, 21.2.
Example 6
Methyl
{4,6-diamino-2-[5-fluoro-1-(2-fluorobenzyl)-1H-pyrazolo[3,4-b]pyrid-
in-3-yl]pyrimidin-5-yl}carbamate phosphate
##STR00039##
[0302] A solution of 100 mg (0.235 mmol) of Example 1 in 2 ml of
1,4-dioxane was prepared in a brown 5 ml glass bottle. In
succession, 2 ml of THF and a solution of 16 ml (0.235 mmol) of 85%
strength phosphoric acid in 0.3 ml of water were added to this
solution, and the solution was stirred at RT until the solvents had
evaporated. Air-drying gave 105 mg (85.4% of theory) of the title
compound.
[0303] PLM (100.times.): crystalline
[0304] DSC: 183.degree. C. (dec., .DELTA.H=65 J/g)
[0305] TGA: 6% weight loss prior to decomposition
[0306] LC-MS (method 3): R.sub.t=0.91 min
[0307] MS (ESIpos): m/z=427 (M+H).sup.+
[0308] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm]=3.57 and
3.62 (2 s, methyl signal obscured by water signal, 3H), 5.79 (s,
2H), 6.22 (br s, 4H), 7.15 (m, 2H), 7.22 (dd, 1H), 7.36 (m, 1H),
7.67 and 7.99 (2 s, 1H), 8.66 (m, 1H), 8.90 (m, 1H).
[0309] .sup.31P-NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm]=-1.1
[0310] Elemental analysis for
C.sub.19H.sub.16F.sub.2N.sub.8O.sub.2+H.sub.3PO.sub.4+2H.sub.2O:
[0311] calculated: % C, 40.72; % H, 4.14; % N, 19.99;
[0312] measured: % C, 40.5; % H, 4.0; % N, 19.5.
Example 7
Methyl
{4,6-diamino-2-[5-fluoro-1-(2-fluorobenzyl)-1H-pyrazolo[3,4-b]pyrid-
in-3-yl]pyrimidin-5-yl}carbamate mesylate
##STR00040##
[0314] A solution of 100 mg (0.235 mmol) of Example 1 in 2 ml of
1,4-dioxane was prepared in a brown 5 ml glass bottle. In
succession, 2 ml of ethanol and a solution of 22.5 mg (0.235 mmol)
of methanesulphonic acid in 0.3 ml of water were added to this
solution, and the solution was stirred at RT until the solvents had
evaporated. Air-drying gave 103 mg (84% of theory) of the title
compound.
[0315] PLM (100.times.): crystalline
[0316] DSC: 154.degree. C. (.DELTA.H=11.7 J/g), 167.degree. C.
(.DELTA.H=-5 J/g), 215.2.degree. C. (dec, .DELTA.H=56.1 J/g)
[0317] TGA: gradual weight loss during the measurement
[0318] LC-MS (method 3): R.sub.t=0.91 min
[0319] MS (ESIpos): m/z=427 (M+H).sup.+
[0320] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm]=2.31 (s,
3H), 3.57 and 3.66 (2 s, 3H), 5.93 (s, 2H), 7.17 (m, 2H), 7.25 (dd,
1H), 7.39 (m, 1H), 7.66 (s br, 4H), 8.06 and 8.34 (2 s, 1H), 8.81
(dd, 1H), 8.83 (s, 1H), 13.0 (br s, 1H).
[0321] Elemental analysis for
C.sub.19H.sub.16F.sub.2N.sub.8O.sub.2+CH.sub.4O.sub.3S+H.sub.2O:
[0322] calculated: % C, 44.44; % H, 4.14; % N, 20.7;
[0323] measured: % C, 44.3; % H, 4.1; % N, 20.2.
Example 8
Methyl
{4,6-diamino-2-[5-fluoro-1-(2-fluorobenzyl)-1H-pyrazolo[3,4-b]pyrid-
in-3-yl]pyrimidin-5-yl}carbamate ethane-1,2-disulphonate
##STR00041##
[0325] A solution of 100 mg (0.235 mmol) of Example 1 in 2 ml of
1,4-dioxane was prepared in a brown 5 ml glass bottle. In
succession, 2 ml of isopropanol and 44.6 mg (0.235 mmol) of
ethane-1,2-disulphonic acid were added to this solution, and the
solution was stirred at RT until the solvents had evaporated.
Air-drying gave 111 mg (73.7% of theory) of the title compound.
[0326] PLM (100.times.): predominantly crystalline
[0327] DSC: 97.degree. C. (dec., .DELTA.H=103 J/g)
[0328] TGA: gradual weight loss during the measurement
[0329] LC-MS (method 3): R.sub.t=0.90 min
[0330] MS (ESIpos): m/z=427 (M+H).sup.+
[0331] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm]=2.66 (s,
4H), 3.57 and 3.66 (2 s, methyl signal obscured by water signal,
3H), 5.93 (s, 2H), 7.17 (m, 2H), 7.25 (m, 1H), 7.39 (m, 1H), 8.05
and 8.35 (2 s, 1H), 8.80 (dd, 1H), 8.84 (s, 1H).
[0332] Elemental analysis for
C.sub.19H.sub.16F.sub.2N.sub.8O.sub.2+C.sub.2H.sub.6O.sub.6S.sub.2+0.25;
H.sub.2O+0.25; C.sub.4H.sub.8O.sub.2:
[0333] calculated: % C, 41.09; % H, 3.84; % N, 17.42;
[0334] measured: % C, 41.2; % H, 4.2; % N, 17.6.
Example 9
Methyl
{4,6-diamino-2-[5-fluoro-1-(2-fluorobenzyl)-1H-pyrazolo[3,4-b]pyrid-
in-3-yl]pyrimidin-5-yl}carbamate maleate
##STR00042##
[0336] A solution of 100 mg (0.235 mmol) of Example 1 in 2 ml of
1,4-dioxane was prepared in a brown 5 ml glass bottle. In
succession, 2 ml of isopropanol and 27.2 mg (0.235 mmol) of maleic
acid were added to this solution, and the solution was stirred at
RT until the solvents had evaporated. Air-drying gave 108 mg (84.9%
of theory) of the title compound.
[0337] PLM (100.times.): crystalline
[0338] DSC: 192.degree. C. (dec., .DELTA.H=173 J/g)
[0339] TGA: 3% weight loss prior to decomposition
[0340] LC-MS (method 3): R.sub.t=0.91 min
[0341] MS (ESIpos): m/z=427 (M+H).sup.+
[0342] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm]=3.56 and
3.64 (2 s, obscured by dioxane signal, 3H), 5.85 (s, 2H), 6.16 (s,
2H), 6.9 (br s, 4H), 7.15 (m, 2H), 7.23 (dd, 1H), 7.37 (m, 1H),
7.85 and 8.13 (2 s, 1H), 8.73 (s, 1H), 8.86 (dd, 1H).
[0343] Elemental analysis for
C.sub.19H.sub.16F.sub.2N.sub.8O.sub.2+C.sub.4H.sub.4O.sub.4+0.5H.sub.2O+0-
.5C.sub.4H.sub.8O.sub.2:
[0344] calculated: % C, 50.42; % H, 4.23; % N, 18.82;
[0345] measured: % C, 50.7; % H, 3.9; % N, 18.8.
Example 10
Methyl
{4,6-diamino-2-[5-fluoro-1-(2-fluorobenzyl)-1H-pyrazolo[3,4-b]pyrid-
in-3-yl]pyrimidin-5-yl}carbamate nitrate
##STR00043##
[0347] A solution of 100 mg (0.235 mmol) of Example 1 in 2 ml of
isopropanol was prepared in a brown 5 ml glass bottle. In
succession, 2 ml of isopropanol and 0.235 .mu.l (0.235 mmol) of 1M
nitric acid were added to this solution, and the solution was
stirred at RT until the solvents had evaporated. Air-drying gave
103 mg (89.7% of theory) of the title compound.
[0348] PLM (100.times.): crystalline
[0349] DSC: 175.degree. C. (dec., .DELTA.H=-224 J/g)
[0350] TGA: 3% weight loss prior to decomposition
[0351] LC-MS (method 3): R.sub.t=0.91 min
[0352] MS (ESIpos): m/z=427 (M+H).sup.+
[0353] .sup.1H NMR (400 MHz, DMSO-d.sub.6): .delta. [ppm]=3.57 and
3.66 (2 s, 3H), 5.93 (s, 2H), 7.16 (m, 2H), 7.25 (dd, 1H), 7.38 (m,
1H), 7.65 (br s, 4H), 8.02 and 8.32 (2 s, 1H), 8.80 (dd, 1H), 8.83
(s, 1H), 13.0 (br s, 1H).
[0354] Elemental analysis for
C.sub.19H.sub.16F.sub.2N.sub.8O.sub.2+HNO.sub.3+0.75H.sub.2O:
[0355] calculated: % C, 45.38; % H, 3.71; % N, 25.07;
[0356] measured: % C, 45.4; % H, 3.7; % N, 25.0.
B. ASSESSMENT OF PHARMACOLOGICAL ACTIVITY
[0357] The pharmacological effect of the compounds according to the
invention can be shown in the following assays:
B-1. Vasorelaxant Effect In Vitro
[0358] Rabbits are stunned by a blow to the neck and exsanguinated.
The aorta is removed, freed from adhering tissue and divided into
rings of a width of 1.5 mm. The rings are placed individually under
an initial tension in 5 ml organ baths with Krebs-Henseleit
solution which is at 37.degree. C., is gassed with carbogen and has
the following composition (in each case mM): sodium chloride 119;
potassium chloride: 4.8; calcium chloride dihydrate: 1; magnesium
sulphate heptahydrate: 1.4; potassium dihydrogenphosphate: 1.2;
sodium bicarbonate: 25; glucose: 10. The force of contraction is
detected with Statham UC2 cells, amplified and digitized via A/D
converters (DAS-1802 HC, Keithley Instruments, Munich) and recorded
in parallel on chart recorders. To produce a contraction,
phenylephrine is added to the bath cumulatively in increasing
concentration. After several control cycles, the substance to be
investigated is added in each further run in increasing dosage in
each case, and the height of the contraction achieved is compared
with the height of the contraction reached in the last preceding
run. The concentration necessary to reduce the height of the
control value by 50% is calculated from this (IC.sub.50 value). The
standard administration volume is 5 .mu.l and the proportion of
DMSO in the bath solution corresponds to 0.1%.
[0359] Representative IC.sub.50 values for the compounds according
to the invention are shown in the table below (Table 1):
TABLE-US-00001 TABLE 1 Example No. IC.sub.50 [nM] 1 958 2 251 3
515
B-2. Effect on a Recombinant Guanvlate Cyclase Reporter Cell
Line
[0360] The cellular activity of the compounds according to the
invention is determined using a recombinant guanylate cyclase
reporter cell line, as described in F. Wunder et al., Anal.
Biochem. 339, 104-112 (2005).
[0361] Representative values (MEC=minimum effective concentration)
for the compounds according to the invention are shown in the table
below (Table 2):
TABLE-US-00002 TABLE 2 Example No. MEC [.mu.M] 1 0.3 2 0.1 3
0.03
B-3. Radiotelemetric Measurement of Blood Pressure on Conscious
Spontaneously Hypertensive Rats
[0362] A commercially available telemetry system from DATA SCIENCES
INTERNATIONAL DSI, USA, is employed for the blood pressure
measurements on conscious rats described below.
[0363] The system consists of 3 main components: [0364] implantable
transmitters (Physiotel.RTM. telemetry transmitter) [0365]
receivers (Physiotel.RTM. receiver) which are linked via a
multiplexer (DSI Data Exchange Matrix) to a [0366] data acquisition
computer.
[0367] The telemetry system makes it possible to continuously
record blood pressure, heart rate and body motions of conscious
animals in their usual habitat.
Animal Material
[0368] The investigations are carried out on adult female
spontaneously hypertensive rats (SHR Okamoto) with a body weight of
>200 g. SHR/NCrl from the Okamoto Kyoto School of Medicine, 1963
were a cross of male Wistar Kyoto rats with highly elevated blood
pressure and female rats having a slightly elevated blood pressure
and at F13 handed over to the U.S. National Institutes of
Health.
[0369] After transmitter implantation, the experimental animals are
housed singly in type 3 Makrolon cages. They have free access to
standard feed and water.
[0370] The day/night rhythm in the experimental laboratory is
changed by the room lighting at 6.00 am and at 7.00 pm.
Transmitter Implantation
[0371] The telemetry transmitters TA11 PA-C40 used are surgically
implanted under aseptic conditions in the experimental animals at
least 14 days before the first experimental use. The animals
instrumented in this way can be employed repeatedly after the wound
has healed and the implant has settled.
[0372] For the implantation, the fasted animals are anaesthetized
with pentobarbital (Nembutal, Sanofi: 50 mg/kg i.p.) and shaved and
disinfected over a large area of their abdomens. After the
abdominal cavity has been opened along the linea alba, the
liquid-filled measuring catheter of the system is inserted into the
descending aorta in the cranial direction above the bifurcation and
fixed with tissue glue (VetBonD.TM., 3M). The transmitter housing
is fixed intraperitoneally to the abdominal wall muscle, and
layered closure of the wound is performed.
[0373] An antibiotic (Tardomyocel COMP, Bayer, 1 ml/kg s.c.) is
administered postoperatively for prophylaxis of infection.
Substances and Solutions
[0374] Unless indicated otherwise, the substances to be
investigated are administered orally by gavage in each case to a
group of animals (n=6). The test substances are dissolved in
suitable solvent mixtures, or suspended in 0.5% strength Tylose,
appropriate for an administration volume of 5 ml/kg of body
weight.
[0375] A solvent-treated group of animals is employed as
control.
Test Procedure
[0376] The telemetry measuring unit present is configured for 24
animals. Each experiment is recorded under an experiment number
(Vyear month day).
[0377] Each of the instrumented rats living in the system is
assigned a separate receiving antenna (1010 Receiver, DSI).
[0378] The implanted transmitters can be activated externally by
means of an incorporated magnetic switch and are switched to
transmission in the run-up to the experiment. The emitted signals
can be detected online by a data acquisition system (Dataquest.TM.
A.R.T. for Windows, DSI) and be appropriately processed. The data
are stored in each case in a file created for this purpose and
bearing the experiment number.
[0379] In the standard procedure, the following are measured for
10-second periods in each case: [0380] systolic blood pressure
(SBP) [0381] diastolic blood pressure (DBP) [0382] mean arterial
pressure (MAP) [0383] heart rate (HR) [0384] activity (ACT).
[0385] The acquisition of measured values is repeated under
computer control at 5-minute intervals. The source data obtained as
absolute value are corrected in the diagram with the currently
measured barometric pressure (Ambient Pressure Reference Monitor;
APR-1) and stored as individual data. Further technical details are
given in the extensive documentation from the manufacturing company
(DSI).
[0386] Unless indicated otherwise, the test substances are
administered at 9.00 am on the day of the experiment. Following the
administration, the parameters described above are measured over 24
hours.
Evaluation
[0387] After the end of the experiment, the acquired individual
data are sorted using the analysis software (Dataquest.TM.
A.R.T..TM. Analysis). The blank value is assumed to be the time 2
hours before administration of the substance, so that the selected
data set includes the period from 7.00 am on the day of the
experiment to 9.00 am on the following day.
[0388] The data are smoothed over a presettable time by
determination of the average (15-minute average) and transferred as
a text file to a storage medium. The measured values presorted and
compressed in this way are transferred into Excel templates and
tabulated. For each day of the experiment, the data obtained are
stored in a dedicated file carrying the number of the experiment.
Results and test protocols are filed in paper form sorted by
numbers.
[0389] Representative values for the compounds according to the
invention are shown in the table below (Table 3):
TABLE-US-00003 TABLE 3 Example 1: Example 2: Dosage Dosage Dosage
0.3 mg/kg 3.0 mg/kg 0.3 mg/kg Vehicle p.o. p.o. Vehicle p.o. hours
after mean blood mean blood mean blood hours after mean blood mean
blood substance pressure pressure pressure substance pressure
pressure administration (mm Hg) (mm Hg) (mm Hg) administration (mm
Hg) (mm Hg) 0 153.6 151.0 149.0 0 149.0 161.3 1 164.5 148.4 129.3 1
158.2 145.7 2 146.7 136.4 111.1 2 142.2 130.5 3 145.4 130.6 106.0 3
149.2 121.5 4 149.6 129.1 107.8 4 152.3 123.1 5 149.9 132.8 109.3 5
155.8 121.6 6 151.6 125.6 106.8 6 147.3 123.8 7 147.6 131.9 110.9 7
147.3 124.4 8 147.5 131.8 109.8 8 149.3 128.7 9 150.8 138.5 114.3 9
151.0 133.7 10 149.8 138.3 114.5 10 152.5 139.2 11 154.0 138.9
115.6 11 150.3 137.9 12 145.3 137.7 118.8 12 146.2 143.0 13 141.1
142.9 120.4 13 143.2 146.0 14 147.8 144.5 122.8 14 146.4 149.2 15
151.0 143.8 125.8 15 150.5 152.3 16 151.3 146.3 131.5 16 145.3
155.5 17 148.8 141.8 124.7 17 143.9 156.3 18 149.2 138.4 129.6 18
150.3 157.3 19 151.2 149.2 135.6 19 147.7 156.9 20 152.6 145.1
135.2 20 153.4 156.3 21 146.3 142.1 129.3 21 148.6 149.3 22 146.3
141.8 128.3 22 153.3 147.1 23 150.3 143.6 130.2 23 151.1 153.1 24
147.4 135.1 130.8 24 154.1 152.3
REFERENCES
[0390] Klaus Witte, Kai Hu, Johanna Swiatek, Claudia Mussig, Georg
Ertl and Bjorn Lemmer: Experimental heart failure in rats: effects
on cardiovascular circadian rhythms and on myocardial
.beta.-adrenergic signaling. Cardiovasc Res 47 (2): 203-405, 2000;
Kozo Okamoto: Spontaneous hypertension in rats. Int Rev Exp Pathol
7: 227-270, 1969; Maarten van den Buuse: Circadian Rhythms of Blood
Pressure, Heart Rate, and Locomotor Activity in Spontaneously
Hypertensive Rats as Measured With Radio-Telemetry. Physiology
& Behavior 55(4): 783-787, 1994
B-4. Determination of Pharmacokinetic Parameters Following
Intravenous and Peroral Administration:
[0391] The pharmacokinetic parameters of the substance are
determined in male CD-1 mice, male Wistar rats and female beagles.
The administration volume is 10 ml/kg for mice, 5 ml/kg for rats
and 0.5 ml/kg for dogs. Intravenous administration is via a
formulation of species-specific plasma/DMSO (99/1) in the case of
mice and rats and via water/PEG400/ethanol (50/40/10) in the case
of dogs. For easier removal of blood, a silicone catheter is
inserted into the right Vena jugularis externa of the rats before
the administration of substance. The surgical intervention takes
place one day prior to the experiment with isofluran anaesthesia
and administration of an analgetic (atropine/rimadyl (3/1) 0.1 ml
s.c.). Substance administration is as i.v. bolus in the case of
mice, as i.v. bolus or via a 15-minute infusion in the case of rats
and via a 15-minute infusion in the case of dogs. Removal of blood
is after 0.033, 0.083, 0.17, 0.5, 1, 2, 3, 4, 6, 7 and 24 hours in
the case of mice and, after a 15-minute infusion, after 0.083,
0.25, 0.28 0.33, 0.42, 0.75, 1, 2, 3, 4, 6, 7 and 24 hours in the
case of dogs and rats and after an i.v. bolus administration, after
0.033, 0.083, 0.17, 0.5, 1, 2, 3, 4, 6, 7 and 24 hours in the case
of rats. For all species, oral administration of the dissolved
substance via gavage is carried out based on a water/PEG400/ethanol
formulation (50/40/10). Here, the removal of blood from rats and
dogs is after 0.083, 0.17, 0.5, 0.75, 1, 2, 3, 4, 6, 7 and 24
hours. The blood is removed into heparinized tubes. The blood
plasma is then obtained by centrifugation; if required, it can be
stored at -20.degree. C. until further processing.
[0392] An internal standard (ZK 228859) is added to the unknown
samples, calibration samples and QCs, and the protein is
precipitated using excess acetonitrile. After addition of an
ammonium acetate buffer (0.01 M, pH 6.8 (Example 1/3) or pH 3.0
(Example 2)) and subsequent vortexing, the mixture is centrifuged
at 1000 g and the supernatant is examined by LC-MS/MS (API 4000, AB
Sciex). Chromatographic separation is carried out on an Agilent
1100-HPLC. The injection volume is 10 .mu.l. The separation column
used is a Phenomenex Luna 5.mu. C8(2) 100A 50.times.2 mm, adjusted
to a temperature of 40.degree. C. For Example 1, a binary mobile
phase gradient at 400 .mu.l/min is used (A: 0.01M ammonium acetate
buffer pH 6.8, B: 0.1% formic acid in acetonitrile): 0 min (90% A),
1 min (90% A), 3.5 min (15% A), 4.5 min (15% A), 4.6 min (90% A), 7
min (90% A). For Example 2, a binary mobile phase gradient at 500
.mu.l/min is used instead (A: 0.01M ammonium acetate buffer pH 3.0,
B: 0.1% formic acid in acetonitrile): 0 min (90% A), 1.5 min (90%
A), 3.5 min (10% A), 4.5 min (10% A), 5 min (90% A), 7 min (90% A).
For Example 3, a binary mobile solvent gradient at 500 .mu.l/min is
used instead (A: 0.01M ammonium acetate buffer pH 6.8, B: 0.1%
formic acid in acetonitrile): 0 min (90% A), 1 min (90% A), 3 min
(10% A), 4 min (10% A), 4.5 min (90% A), 6 min (90% A). The
temperature of the Turbo V ion source is 500.degree. C. The
following MS instrument parameters are used: curtain gas 20 units
(Example 1), 16 units (Example 2) or 15 units (Example 3), ion
spray voltage 5 kV (Example 1/2) or 4.5 units (Example 3), gas 1 35
units (Example 1/3) or 25 units (Example 2), gas 2 30 units, CAD
gas 4 units (Example 1/3) or 3 units (Example 2). The substances
are quantified by peak heights or areas using extracted ion
chromatograms of specific MRM experiments.
[0393] The plasma concentration/time plots determined are used to
calculate the pharmacokinetic parameters such as AUC, C.sub.max,
MRT (mean residence time), t.sub.1/2 (half life) and CL (clearance)
employing the validated pharmacokinetic calculation programs KinEx
(Vers. 2.5 and 3).
[0394] As the substance quantification is carried out in plasma, it
is necessary to determine the blood/plasma distribution of the
substance to be able to adjust the pharamacokinetic parameters in
an appropriate manner. To this end, a defined amount of substance
is incubated in heparinized whole blood of the species in question
in a rocking roller mixer for 20 min. After centrifugation at 1000
g, the plasma concentration is measured (see above) and determined
by calculating the quotient of the c.sub.b/c.sub.p values.
[0395] Following intravenous administration of 0.3 mg/kg of the
compounds according to the invention in rats, the following values
were recorded:
TABLE-US-00004 Example 1.* 2.** 3.** AUC.sub.norm [kg h/l] 4.36
1.79 1.36 CL.sub.blood [lh/kg] 0.29 0.53 1.02 MRT [h] 4.1 2.3 2.3
t.sub.1/2 [h] 3.4 1.7 1.9 *15-minute infusion **i.v. bolus
administration
B-5. Safety Profile
[0396] The substances according to the invention show a
surprisingly favourable safety profile in vivo which was
established by non-clinical safety studies according to OECD (OECD
guidelines for testing of chemicals, No. 407) and ICH (3BS2A)
guidelines.
C. WORKING EXAMPLES FOR PHARMACEUTICAL COMPOSITIONS
[0397] The compounds according to the invention can be converted to
pharmaceutical formulations as follows
Tablet:
Composition:
[0398] 100 mg of the compound according to the invention, 50 mg of
lactose (monohydrate), 50 mg of maize starch (native), 10 mg of
polyvinylpyrrolidone (PVP 25) (BASF, Ludwigshafen, Germany) and 2
mg of magnesium stearate.
[0399] Tablet weight 212 mg, diameter 8 mm, radius of curvature 12
mm.
Preparation:
[0400] The mixture of the compound according to the invention,
lactose and starch is granulated with a 5% solution (w/w) of the
PVP in water. After drying, the granules are mixed with the
magnesium stearate for 5 minutes. This mixture is pressed with a
conventional tableting press (for tablet dimensions see above). The
guide value used for the pressing is a pressing force of 15 kN.
Suspension for Oral Administration:
Composition:
[0401] 1000 mg of the compound according to the invention, 1000 mg
of ethanol (96%), 400 mg of Rhodigel.RTM. (xanthan gum from FMC,
Pennsylvania, USA) and 99 g of water.
[0402] A single dose of 100 mg of the compound according to the
invention corresponds to 10 ml of oral suspension.
Preparation:
[0403] The Rhodigel is suspended in ethanol and the compound
according to the invention is added to the suspension. The water is
added while stirring. The mixture is stirred for approx. 6 h until
swelling of the Rhodigel has ended.
Solution for Oral Administration:
Composition:
[0404] 500 mg of the compound according to the invention, 2.5 g of
polysorbate and 97 g of polyethylene glycol 400. A single dose of
100 mg of the compound according to the invention corresponds to 20
g of oral solution.
Preparation:
[0405] The compound according to the invention is suspended in the
mixture of polyethylene glycol and polysorbate while stirring. The
stirring operation is continued until dissolution of the compound
according to the invention is complete.
i.v. Solution:
[0406] The compound according to the invention is dissolved in a
concentration below the saturation solubility in a physiologically
acceptable solvent (e.g. isotonic saline, glucose solution 5%
and/or PEG 400 solution 30%). The solution is subjected to sterile
filtration and dispensed into sterile and pyrogen-free injection
vessels.
* * * * *