U.S. patent number RE38,624 [Application Number 10/176,435] was granted by the patent office on 2004-10-12 for hydroxyindoles, their use as inhibitors of phosphodiesterase 4 and process for their preparation.
This patent grant is currently assigned to Elbion AG. Invention is credited to Ute Egerland, Sabine Heer, Norbert Hofgen, Thomas Kronbach, Hildegard Kuss, Degenhard Marx, Emmanuel Polymeropoulos, Stefan Szelenyi.
United States Patent |
RE38,624 |
Hofgen , et al. |
October 12, 2004 |
Hydroxyindoles, their use as inhibitors of phosphodiesterase 4 and
process for their preparation
Abstract
The invention relates to new hydroxyindoles of the Formula,
##STR1## their use as inhibitors of phosphodiesterase 4 and
processes for their preparation.
Inventors: |
Hofgen; Norbert
(Ottendirk-Okrilla, DE), Egerland; Ute (Radebavl,
DE), Kuss; Hildegard (Dresden, DE), Marx;
Degenhard (Radolfzell-Markelfingen, DE), Szelenyi;
Stefan (Schwaig, DE), Kronbach; Thomas (Radebeul,
DE), Polymeropoulos; Emmanuel (Frankfurt,
DE), Heer; Sabine (Radebeul, DE) |
Assignee: |
Elbion AG (Radebeul,
DE)
|
Family
ID: |
26045814 |
Appl.
No.: |
10/176,435 |
Filed: |
September 19, 2002 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
Issue Date |
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Reissue of: |
300973 |
Apr 28, 1999 |
06251923 |
Jun 26, 2001 |
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Foreign Application Priority Data
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Apr 28, 1998 [DE] |
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198 18 964 |
Apr 17, 1999 [DE] |
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199 17 504 |
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Current U.S.
Class: |
514/339;
546/278.1 |
Current CPC
Class: |
A61P
21/00 (20180101); A61P 19/08 (20180101); A61P
1/00 (20180101); A61P 25/24 (20180101); A61P
33/06 (20180101); A61P 37/00 (20180101); A61P
25/02 (20180101); A61P 31/04 (20180101); A61P
9/10 (20180101); A61P 11/08 (20180101); C07D
209/24 (20130101); A61P 3/10 (20180101); A61P
25/28 (20180101); A61P 9/00 (20180101); A61P
37/06 (20180101); A61P 11/02 (20180101); A61P
17/04 (20180101); A61P 43/00 (20180101); A61P
11/00 (20180101); A61P 25/36 (20180101); A61P
17/02 (20180101); A61P 19/00 (20180101); C07D
401/12 (20130101); A61P 13/10 (20180101); A61P
1/04 (20180101); A61P 37/02 (20180101); A61P
9/08 (20180101); A61P 19/02 (20180101); A61P
17/00 (20180101); A61P 25/16 (20180101); A61P
11/06 (20180101); A61P 13/04 (20180101); A61P
25/00 (20180101); A61P 13/12 (20180101); C07D
209/22 (20130101); A61P 7/12 (20180101); A61P
29/00 (20180101); A61P 13/08 (20180101); A61P
17/06 (20180101); A61P 27/14 (20180101); A61P
31/18 (20180101) |
Current International
Class: |
C07D
401/06 (20060101); C07D 401/00 (20060101); A61K
31/4439 (20060101); A61K 31/4427 (20060101); A61K
031/443 (); C07D 401/06 () |
Field of
Search: |
;514/339 ;546/278.1 |
References Cited
[Referenced By]
U.S. Patent Documents
Foreign Patent Documents
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2057181 |
|
Jun 1992 |
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CA |
|
2195850 |
|
Feb 1996 |
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CA |
|
2215013 |
|
Mar 1998 |
|
CA |
|
195 11 916 |
|
Feb 1996 |
|
DE |
|
773-024A 2 |
|
May 1997 |
|
EP |
|
94/12461 |
|
Jun 1994 |
|
WO |
|
95/14667 |
|
Jun 1995 |
|
WO |
|
95/24408 |
|
Sep 1995 |
|
WO |
|
97/48697 |
|
Dec 1997 |
|
WO |
|
98/02151 |
|
Jan 1998 |
|
WO |
|
98/08818 |
|
Mar 1998 |
|
WO |
|
Other References
CA 108:94380, Nakao et al, 1998.* .
The Merck Index, 11th ed., pp 786-787.* .
Thomas Hoffman et al. Absence of modulation of monokine, etc., CA
(1991) 114:240174 a.* .
Linda S. Brady et al., 1,1'-ethylidenebis [L-tryptophan], etc., CA
(1994) 121:81303t.* .
Toray Ind., "Preparation of tererocyclyl compounds, etc.",
CA(1997), No. 8, 127:108841e.* .
Brian L. Williamson, "Online HPLC-tandem mass spectrometry, etc.",
CA No. 16 (1997) 127:219990x.* .
C. Brideau et al, "Pharmacology of MK-0591, etc.", CA (1992)
117:22596r.* .
H. Sidransky et al, Studies with 1,1'-ethylbis (tryptophan), a
contaminant, etc., CA (1994), 121:2861x..
|
Primary Examiner: Fan; Jane
Attorney, Agent or Firm: Fulbright & Jaworski L.L.P.
Claims
We claim:
1. A compound of the formula (I) ##STR4##
wherein R.sup.1 is a straight or branched .[.C.sub.1-3 .].
.Iadd.C.sub.1-12 .Iaddend.alkyl optionally substituted with .Iadd.a
.Iaddend.phenyl .Iadd.radical.Iaddend., or C.sub.3-8 cycloalkyl
.[.residue.]. .Iadd.radical .Iaddend.wherein the phenyl
.[.residue.]. .Iadd.radical .Iaddend.is optionally substituted with
a halo, nitro, hydroxy, C.sub.1-4 alkyl, C.sub.1-4 alkoxy, or COOH
.[.residue.]. .Iadd.radical.Iaddend.; R.sup.2 and R.sup.3 are each
independently of each other hydrogen, or an --OH .[.residue.].
.Iadd.radical .Iaddend.where at least one of R.sup.2 and R.sup.3
are --OH; R.sup.5 is a pyridyl .[.residue.]. .Iadd.radical
.Iaddend.disubstituted with a halogen .[.residue.].
.Iadd.radical.Iaddend.; and A is a bond, C.ident.O, or a CHOH
.[.residue.]. .Iadd.radical.Iaddend., or the pharmaceutically
acceptable salt thereof.
2. The compound of claim 1, wherein the compound is a
pharmaceutically acceptable salt of an organic or inorganic acid,
or of an organic or inorganic base, or a quaternary ammonium salt
from the quaternization of a tertiary amine.
3. The compound of claim 1, having an asymetric carbon atom by
being the L or the D form, or a D,L mixture, and when in a
diastereoisomeric form..[.
4. A compound of claim 1, being one of the following compounds:
N-(3,5-dichloropyridin-4-yl)-2-[1-(4-fluorobenzyl)-5-hydroxyindol-3-yl]-2-o
xoacetamide;
N-(3,5-dichloropyridin-4-yl)-2-[1-(4-fluorobenzyl)-5-hydroxyindol-3-yl]-2-o
xoacetamide Na salt;
N-(3,5-dichloropyridin-4-yl)-2-[1-(4-fluorobenzyl)-5-hydroxyindol-3-yl]-2-h
ydroxyacetamide;
N-(pyridin-4-yl)-2-[1-2,6-difluorobenzyl)-5-hydroxyindol-3-yl]-2-oxyacetami
de;
N-(3,5-dichloropyridin-4-yl)-2-[1-(2,6-difluorobenzyl)-5-hydroxyindol-3-yl]
-2-oxoacetamide;
N-(3,5-dichloropyridin-4-yl)-2-[1-(3-nitrobenzyl)-5-hydroxyindol-3-yl]-2-ox
oacetamide Na salt;
N-(3,5-dichloropyridin-4-yl)-2-(1-propyl-5-hydroxyindol-3-yl)-2-oxyacetamid
e;
N-(3,5-dichloropyridin-4-yl)-2-(1-isopropyl-5-hydroxyindol-3-yl)-2-oxoaceta
mide;
N-(3,5-dichloropyridin-4-yl)-2-(1-cyclopentylmethyl-5-hydroxyindol-3-yl)-2-
oxoacetamide;
N-(2,6-dichlorophenyl)-2-[1-(4-fluorobenzyl)-5-hydroxyindol-3-yl]-2-oxoacet
amide;
N-(2,6-dichloro-4-trifluoromethylphenyl)-2-[1-(4-fluorobenzyl)-5-hydroxyind
ol-3-yl)-2-oxoacetamide;
N-(2,6-dichloro-4-trifluoromethoxylphenyl)-2-[1-(4-fluorobenzyl)-5-hydroxyi
ndol-3-yl)-2-oxoacetamide;
N-(3,5-dichloropyridin-4-yl)-2-[1-(4-fluorobenzyl)-6-hydroxyindol-3-yl]-2-o
xoacetamide;
N-(3,5-dichloropyridin-4-yl)-5-hydroxy-1-(4-methoxybenzyl)indole-3-carboxam
ide..].
5. A process for inhibiting TNF.alpha. by administering to a
patient in need therefor an effective amount of the compound of
claim 1.
6. A process for inhibiting TNF.alpha. by administering to a
patient in need therefor an effective amount of the compound of
claim .[.4.]. .Iadd.2.Iaddend..
7. A process for inhibiting phosphodiesterase 4 by administering to
a patient in need therefor an effective amount of the compound of
claim 1.
8. A process for inhibiting phosphodiesterase 4 by administering to
a patient in need therefor an effective amount of the compound of
claim .[.4.]. .Iadd.2.Iaddend..
9. A process for treating an eosinophil-related condition by
administering to a patient in need therefor an effective amount of
the compound of claim 1.
10. A process for treating an eosinophil-related condition by
administering to a patient in need therefor an effective amount of
the compound of claim .[.4.]. .Iadd.2.Iaddend..
11. A process for treating a chronic obstructive pulmonary disease,
which comprises administering to a patient in need therefor an
effective amount of a compound of claim 1.
12. A process for treating a chronic obstructive pulmonary disease,
which comprises administering to a patient in need therefor an
effective amount of a compound of claim .[.4.].
.Iadd.2.Iaddend..
13. A pharmaceutical preparation which comprises a therapeutically
effective amount of the compound of claim 1, together with one or
more of a pharmaceutically acceptable carrier, diluent, and
auxiliary ingredient.
14. A process for preparing the pharmaceutical preparation of claim
7, which comprises preparing a pharmaceutically acceptable dosage
form from a compound of formula (I)and from one or more of a
pharmaceutically acceptable carrier, diluent, and auxiliary
ingredient.
Description
FIELD OF THE INVENTION
The present invention relates to novel, substituted hydroxyindoles
processes for their preparation, pharmaceutical preparations
containing these compounds, and a method for the use of these
compounds which are phosphodiesterase 4 inhibitors, as active
compounds for the treatment of disorders which can be affected by
inhibition of phosphodiesterase 4 activity in immunocompetent cells
(e.g. macrophages and lymphocytes).
BACKGROUND
The activation of cell membrane receptors by transmitters leads to
the activation of the second messenger system. Adenylate cyclase
synthesizes active cyclic AMP (cAMP) or cyclic GMP (cGMP) from AMP
and GMP. These lead, for example, to relaxation in smooth muscle
cells or to inhibition of mediator release or synthesis in
inflammatory cells. The breakdown of the second messenger cAMP and
cGMP is carried out by the phosphodiesterases (PDE). To date, 7
families of PDE enzymes (PDE1-7) are known, which differ by their
substrate specificity (cAMP, cGMP or both) and the dependence on
other substrates (e.g. calmodulin). These isoenzymes have different
functions in the body and are prominent to different extents in the
individual cell types (Beave J A, Conti M and Heaslip R J, Multiple
cyclic nucleotide phosphodiesterases, Mol. Pharmacol. 1994, 46:
399-405; Hall I P, Isoenzyme selective phosphodiesterase
inhibitors; potential clinical uses, Br. J. clin. Pharmacol. 1993,
35: 1-7). As a result of inhibition of the various PDE isoenzyme
types, there is an accumulation of cAMP or cGMP in the cells, which
can be therapeutically utilized (Torphy T J, Livi G P, Christensen
S B, Novel Phosphodiesterase Inhibitors for the Therapy of Asthma,
Drug News and Perspectives 1993, 6: 203-214).
In the cells important for allergic inflammation (lymphocytes, mast
cells, eosinophilic granulocytes, macrophages), the prevailing PDE
isoenzyme is of type 4 (Torphy, J T. and Undem, B. J.,
Phosphodiesterase inhibitors: new opportunities for the treatment
of asthma, Thorax 1991, 46: 512-523). The inhibition of PDE 4 by
suitable inhibitors is therefore considered as an important
starting point for the therapy of a large number of allergically
induced disorders (Schudt Ch, Dent G, Rabe K, Phosphodiesterase
Inhibitors, Academic Press London 1996).
An important property of phosphodiesterase 4 inhibitors is the
inhibition of the release of tumour necrosis factor .alpha.
(TNF.alpha.) from inflammatory cells. TNF.alpha. is an important
pro-inflammatory cytokine, which affects a large number of
biological processes. TNF.alpha. is released, for example, from
activated macrophages, activated T lymphocytes, mast cells,
basophils, fibroblasts, endothelial cells and astrocytes in the
brain. It has a self-activating effect on neutrophils, eosinophils,
fibroblasts and endothelial cells, as a result of which various
tissue-destroying mediators are released. In monocytes, macrophages
and T lymphocytes, TNF.alpha. brings about the increased production
of further pro-inflammatory cytokines such as GM-CSF
(granulocyte-macrophage colony-stimulating factor) or
interleukin-8. TNF.alpha. plays a central part due to its
inflammation-promoting and catabolic action in a large number of
disorders, such as inflammation of the airways, inflammation of the
joints, endotoxic shock, tissue rejection, AIDS and numerous other
immunological disorders. Inhibitors of phosphodiesterase 4 are thus
also suitable for the therapy of disorders of this type which are
associated with TNF.alpha..
Chronic obstructive pulmonary diseases (COPD) are widespread in the
population and also have great economic importance. Thus COPD
diseases cause about 10-15% of all illness costs in the developed
countries and about 25% of all cases of death in the USA are to be
attributed to this cause (Norman P.: COPD: New developments and
therapeutic opportunities, Drug News Perspect. 11 (7), 431-437,
1998), however the patients at the time of death are usually over
55 years old (Nolte D.: Chronische Bronchitis--eine Volkskrankheit
multifaktorieller Genese. Atemw.-Lungenkrkh. [Chronic bronchitis--a
widespread disease of multifactorial origin]. 20 (5), 260-267,
1994). The WHO estimates that COPD will be the third most frequent
cause of death within the next 20 years.
The syndrome of chronic obstructive lung diseases (COPD) summarizes
various syndromes of chronic bronchitis with the symptoms coughing
and expectoration and progressive and irreversible impairment of
lung function (exhalation is particularly affected). The course of
the disease is episodic and often complicated by bacterial
infections (Rennard S. I.: COPD: Overview of definitions,
Epidemiology, and factors influencing its development. Chest, 113
(4) Suppl., 235S-241S, 1998). In the course of the disease, the
lung function continuously decreases, the lungs become increasingly
emphysematous and the respiratory distress of the patients is
obvious. This disease clearly adversely affects the quality of life
of the patients (dyspnoea, low exercise tolerance) and
significantly reduces their life expectancy. The main risk factor
besides environmental factors is smoking (Kummer F.: Asthma und
COPD. Atemw.-Lungenkrkh. 20 (5), 299-302, 1994; Rennard S. I.:
COPD: Overview of definitions, Epidemiology, and factors
influencing its development. Chest, 113 (4) Suppl., 235S-241S,
1998) and therefore men are clearly more often affected than women.
As a result of the change in living habits and the increase in the
number of smokers, this picture, however, will change in
future.
The current therapy aims only at the alleviation of the symptoms,
without causally intervening in the progression of the disease. The
use of long-acting Beta2 agonists (e.g. salmeterol) possibly in
combination with muscarinergic antagonists (e.g. ipratropium)
improves the lung function by bronchodilatation and is employed
routinely (Norman P.: COPD: New developments and therapeutic
opportunities, Drugs News Perspect. 11 (7), 431-437, 1998). A large
part in the COPD episodes is played by bacterial infections, which
have to be treated with antibiotics (Wilson R.: The role of
infection in COPD, Chest, 113 (4) Suppl., 242S-248S, 1998; Grossman
R. F.: The value of antibiotics and the outcomes of antibiotic
therapy in exacerbations of COPD. Chest, 113 (4) Suppl., 249S-255S,
1998). The therapy of this disease is unsatisfactory as yet,
particularly with respect to the continuous decrease in lung
function. New therapeutic approaches which affect inflammatory
mediators, proteases or adhesion molecules could be very promising
(Barnes P. J.: Chronic obstructive disease: new opportunities for
drug development, TiPS 10 (19), 415-423, 1998).
Independently of the bacterial infections complicating the disease,
a chronic inflammation which is dominated by neutrophilic
granulocytes is found in the bronchi. The mediators and enzymes
released by neutrophilic granulocytes, inter alia, have been held
responsible for the structural changes observed in the airways
(emphysema). The inhibition of the activity of the neutrophilic
granulocytes is thus a rational approach to prevent or to slow down
progression of COPD (impairment of lung function parameters). An
important stimulus for the activation of the granulocytes is the
pro-inflammatory cytokine TNFO.alpha. (tumour necrosis factor).
Thus it is known that TNF.alpha. stimulates the formation of oxygen
radicals by neutrophilic granulocytes (Jersmann, H. P. A.; Rathjen,
D. A. and Ferrante A.: Enhancement of LPS-induced neutrophil oxygen
radical production by TNF.alpha., Infection and Immunity, 4,
1744-1747, 1998). PDE4 inhibitors can very effectively inhibit the
release of TNF.alpha. from a large number of cells and thus
suppress the activity of the neutrophilic granulocytes. The
non-specific PDE inhibitor pentoxifylline is able to inhibit both
the formation of oxygen radicals and the phagocytosability of
neutrophilic granulocytes (Wenisch, C.; Zedtwitz-Liebenstein, K.;
Parschalk, B. and Graninger W.: Effect of pentoxifylline in vitro
on neutrophil reactive oxygen production and phagocytic ability
assessed by flow cytometry, Clin. Drug. Invest., 13(2):99-104,
1997).
Various PDE 4 inhibitors are already known. As a matter of
priority, these are xanthine derivatives, rolipram analogues or
nitraquazone derivatives (general survey in: Karlsson J-A, Aldos D,
Phosphodiesterase 4 inhibitors for the treatment of asthma, Exp.
Opin. Ther. Patents 1997, 7: 989-1003). Until now, it was not
possible to use any of these compounds clinically. It had to be
established that the known PDE 4 inhibitors also have various
side-effects such as nausea and emesis, which it was not possible
to suppress adequately until now. The discovery of new PDE 4
inhibitors with better therapeutic breadth is therefore
necessary.
Although indoles have been playing an important part for many years
in the development of new active compounds for various indications,
until now hydroxyindoles were completely unknown as inhibitors of
PDE 4.
DESCRIPTION OF THE INVENTION
The invention relates to substituted hydroxyindoles of the Formula
##STR2##
and their pharmaceutically acceptable salts, wherein
R.sup.1, R.sup.5 are independently of each other
(i) a .[.C.sub.1-2 .]. .Iadd.C.sub.1-12 .Iaddend.alkyl,
straight-chain or branched-chain, optionally mono- or
.[.polysubstituted.]. .Iadd.polysubstituted .Iaddend.by --OH, --SH,
--NH.sub.2, --NHC.sub.1-6 alkyl, --N(C.sub.1-6 alkyl).sub.2,
--NHC.sub.6-14 aryl, --N(C.sub.6-14 aryl).sub.2, --N(C.sub.1-6
alkyl)(C.sub.6-14 aryl), --NHCOR.sup.6, --NO.sub.2, --CN, --F,
--Cl, --Br, --I, --O--C.sub.1-6 alkyl, --O--C.sub.6-14 aryl,
--O(CO)R.sup.6, --S--C.sub.1-6 alkyl, --S--C.sub.6-14 aryl,
--SOR.sup.6, --SO.sub.3 H, --SO.sub.2 R.sup.6, --OSO.sub.2
C.sub.1-6 alkyl, --OSO.sub.2 C.sub.6-14 aryl, --(CS)R.sup.6,
--COOH, --(CO)R.sup.6, mono-, bi- or tricyclic saturated or mono-
or polyunsaturated carbocycles having from 3 to 14 ring members,
mono-, bi- or tricyclic saturated or mono- or polyunsaturated
heterocycles having from 5 to 15 ring members and from 1 to 6
heteroatoms, which are suitably N, O and S, where the C.sub.6-4
aryl groups and the included carbocyclic and heterocyclic
substituents can optionally be mono- or polysubstituted by
R.sup.4,
(ii) --C.sub.2-12 alkenyl, mono- or polyunsaturated, straight-chain
or branched-chain, optionally mono- or polysubstituted by --OH,
--SH, --NH.sub.2, --NHC.sub.1-6 alkyl, --N(C.sub.1-6 alkyl).sub.2,
--NHC.sub.6-14 aryl, --N(C.sub.6-14 aryl).sub.2, --N(C.sub.1-6
alkyl)(C.sub.6-14 aryl), --NHCOR.sup.6, --NO.sub.2, --CN, --F,
--Cl, --Br, --I, --O--C.sub.1-6 alkyl, --O--C.sub.6-14 aryl,
--O(CO)R.sup.6, --S--C.sub.1-6 alkyl, --S--C.sub.6-14 aryl,
--SOR.sup.6, --SO3H, --SO.sub.2 R.sup.6, --OSO.sub.2 C.sub.1-6
alkyl, --OSO.sub.2 C.sub.6-14 aryl, --(CS)R.sup.6, --COOH,
--(CO)R.sup.6, mono-, bi- or tricyclic saturated or mono- or
polyunsaturated carbocycles having from 3 to 14 ring members,
mono-, bi- or tricyclic saturated or mono- or polyunsaturated
heterocycles having from 5 to 15 ring members and from 1 to 6
heteroatoms, which are suitably N, O and S, where the C.sub.6-14
aryl groups and the included carbocyclic and heterocyclic
substituents for their part can optionally be mono- or
polysubstituted by R.sup.4,
(iii) mono-, bi- or tricyclic saturated or mono- or polyunsaturated
carbocycles having from 3 to 14 ring members, optionally mono- or
polysubstituted by --OH, --SH, --NH.sub.2, --NHC.sub.1-6 alkyl,
--N(C.sub.1-6 alkyl).sub.2, --NHC.sub.6-14 aryl, --N(C.sub.6-14
aryl).sub.2, --N(C.sub.1-6 alkyl)(C.sub.6-14 aryl), --NHCOR.sup.6,
--NO.sub.2, --CN, --F, --Cl, --Br, --I, --O--C.sub.1-6 alkyl,
--O--C.sub.6-14 aryl, --O(CO)R.sup.6, --S--C.sub.1-6 alkyl,
--S--C.sub.6-14 aryl, --SOR.sup.6, --SO.sub.3 H, --SO.sub.2
R.sup.6, --OSO.sub.2 C.sub.1-6 alkyl, --OSO.sub.2 C.sub.6-14 aryl,
--(CS)R.sup.6, --COOH, --(CO)R.sup.6, mono-, bi- or tricyclic
saturated or mono- or polyunsaturated carbocycles having from 3 to
14 ring members, mono-, bi- or tricyclic saturated or mono- or
polyunsaturated heterocycles having from 5 to 15 ring members and
from 1 to 6 heteroatoms, which are suitably N, O and S, where the
C.sub.6-14 aryl groups and the included carbocyclic and
heterocyclic substituents can optionally be mono- or
polysubstituted by R.sup.4,
(iv) mono-, bi- or tricyclic saturated or mono- or polyunsaturated
heterocycles having from 5 to 15 ring members and from 1 to 6
heteroatoms, which are suitably N, O and S, optionally mono- or
polysubstituted by --OH, --SH, --NH.sub.2, --NHC.sub.1-6 alkyl,
--N(C.sub.1-6 alkyl).sub.2, --NHC.sub.6-14 aryl, --N(C.sub.6-14
aryl).sub.2, --N(C.sub.1-6 alkyl)(C.sub.6-14 aryl), --NHCOR.sup.6,
--NO.sub.2, --CN, --F, --Cl, --Br, --I, --O--C.sub.1-6 alkyl,
--O--C.sub.6-14 aryl, --O(CO)R.sup.6, --S--C.sub.1-6 alkyl,
--S--C.sub.6-14 aryl, --SOR.sup.6, --SO.sub.3 H, --SO.sub.2
R.sup.6, --OSO.sub.2 C.sub.1-6 alkyl, --OSO.sub.2 C.sub.6-14 aryl,
--(CS)R.sup.6, --COOH, --(CO)R.sup.6, mono-, bi- or tricyclic
saturated or mono- or polyunsaturated carbocycles having from 3 to
14 ring members, mono-, bi- or tricyclic saturated or mono- or
polyunsaturated heterocycles having from 5 to 15 ring members and
from 1 to 6 heteroatoms, which are suitably N, O and S, where the
C.sub.6-14 aryl groups and the included carbocyclic and
heterocyclic substituents for their part can be optionally mono- or
polysubstituted by R.sup.4, -carbo- or heterocyclic saturated or
mono- or polyunsaturated spirocycles having from 3 to 10 ring
members, where heterocyclic systems contains from 1 to 6
heteroatoms, which are suitably N, O and S, optionally mono- or
polysubstituted by --OH, --SH, --NH.sub.2, --NHC.sub.1-6 alkyl,
N(C.sub.1-6 alkyl).sub.2, --NHC.sub.6-14 aryl, --N(C.sub.6-14
aryl).sub.2, --N(C.sub.1-6 alkyl) (C.sub.6-14 aryl, --NHCOR.sup.6,
--NO.sub.2, --CN, --F, --Cl, --Br, --I, --O--C.sub.1-6 alkyl,
--O--C.sub.6-14 aryl, --O(CO)R.sup.6, --S--C.sub.1-6 alkyl,
--S--C.sub.6-14 aryl, --SOR.sup.6, --SO3H, --SO.sub.2 R.sup.6,
OSO.sub.2 C.sub.1-6 alkyl, --OSO.sub.2 C.sub.6-14 aryl,
--(CS)R.sup.6, --COOH, --(CO)R.sup.6, mono-, bi- or tricyclic
saturated or mono- or polyunsaturated carbocycles having from 3 to
14 ring members, mono-, bi- or tricyclic saturated or mono- or
polyunsaturated heterocycles having from 5 to 15 ring members and
from 1 to 6 heteroatoms, which are suitably N, O and S, where the
C.sub.6-14 aryl groups and the included carbocyclic and
heterocyclic substituents can optionally be mono- or
polysubstituted by R.sup.4,
R.sup.2, R.sup.3 are hydrogen or --OH, where at least one of the
two substituents must be --OH;
R.sup.4 is --H, --OH, --SH, --NH.sub.2, --NHC.sub.1-6 alkyl,
--N(C.sub.1-6 alkyl).sub.2, --NHC.sub.6-14 aryl, --N(C.sub.6-14
aryl).sub.2, --N(C.sub.1-6 alkyl)(C.sub.6-14 aryl), --NHCOR.sup.6,
--NO.sub.2, --CN, --COOH, --(CO)R.sup.6, --(CS)R.sup.6, --F, --Cl,
--Br, --I, --O--C.sub.1-6 alkyl, --O--C.sub.6-14 aryl,
--O(CO)R.sup.6, --S--C.sub.1-6 alkyl, --S--C.sub.6-14 aryl,
--SOR.sup.6, --SO.sub.2 R.sup.6
R.sup.6 is --H, --NH.sub.2, --NHC.sub.1-6 alkyl, --N(C.sub.1-6
alkyl).sub.2, --NHC.sub.6-14 aryl, --N(C.sub.6-14 aryl).sub.2,
N(C.sub.1-6 alky)(C.sub.6-14 aryl), --O--C.sub.1-6 alkyl,
--O--C.sub.6-14 aryl, --S--C.sub.1-6 alkyl, --S--C.sub.6-14 aryl,
--C.sub.1-12 alkyl, straight-chain or branched-chain, --C.sub.2-12
alkenyl, mono- or polyunsaturated, straight-chain or
branched-chain, -mono-, bi- or tricyclic saturated or mono- or
polyunsaturated carbocycles having from 3 to 14 ring members,
-mono-, bi- or tricyclic saturated or mono- or polyunsaturated
heterocycles having from 5 to 15 ring members and from 1 to 6
heteroatoms, which are suitably N, O and S;
A is either a bond, or --CH2).sub.m --, --(CH2).sub.m
--(CH.ident.CH).sub.n --(CH.sub.2).sub.p --, --(CHOZ).sub.m --,
--(C.ident.O)--, --(C.ident.S)--, --(C.ident.N-Z)--, --O--, --S--,
--NZ--, where m and p are cardinal numbers from 0 to 3 and n is a
cardinal number from 0 to 2,
Z is H, or a C.sub.1-12 alkyl, straight-chain or branched-chain,
C.sub.2-12 alkenyl, mono- or polyunsaturated, straight-chain or
branched-chain, mono-, bi- or tricyclic saturated or mono- or
polyunsaturated carbocycles having from 3 to 14 ring members,
mono-, bi- or tricyclic saturated or mono- or polyun-saturated
heterocycles having from 5 to 15 ring members and from 1 to 6
heteroatoms, which are suitably N, O and S;
B is either carbon or sulfur, or --(S.ident.O)--;
D is oxygen, sulfur, CH.sub.2 or N-Z, where D can only be S or
CH.sub.2 if B is carbon;
E is a bond, or (CH2).sub.m --, --O--, --S--, --(N-Z)--, where m
and Z have the same meanings as above.
The most suitable compounds of Formula (1) include
N-(3,5-dichloropyridin-4-yl)-2-[1-(4-fluorobenzyl)-5-hydroxyindol-3-yl]-2-o
xoacetamide;
N-(3,5-dichloropyridin-4-yl)-2-[1-(4-fluorobenzyl)-5-hydroxyindol-3-yl]-2-o
xoacetamide Na salt;
N-(3,5-dichloropyridin-4-yl)-2-[1-(4-fluorobenzyl)-5-hydroxyindol-3-yl]-2-h
ydroxyacetamide;
N-(pyridin-4-yl)-2-[1-2,6-difluorobenzyl)-5-hydroxyindol-3-yl]-2-oxoacetami
de;
N-(3,5-dichloropyridin-4-yl)-2-[1-(2,6-difluorobenzyl)-5-hydroxyindol-3-yl]
-2-oxoacetamide;
N-(3,5-dichloropyridin-4-yl)-2-[1-(3-nitrobenzyl)-5-hydroxyindol-3-yl]-2-ox
oacetamide Na salt;
N-(3,5-dichloropyridin-4-yl)-2-(1-propyl-5-hydroxyindol-3-yl)-2-1-oxoacetam
ide;
N-(3,5-dichloropyridin-4-yl)-2-(1-isopropyl-5-hydroxyindol-3-yl)-2-oxoaceta
mide;
N-(3,5-dichloropyridin-4-yl)-2-(1-cyclopentylmethyl-5-hydroxyindol-3-yl)-2-
oxoacetamide;
N-(2,6-dichlorophenyl)-2-[1-(4-fluorobenzyl)-5-hydroxyindol-3-yl]-2-oxoacet
amide;
N-(2,6-dichloro-4-trifluoromethylphenyl)-2-[1-(4-fluorobenzyl)-5-hydroxyind
ol-3-yl)-2-oxoacetamide;
N-(2,6-dichloro-4-trifluoromethoxylphenyl)-2-[1-(4-fluorobenzyl)-5-hydroxyi
ndol-3-yl)-2-oxoacetamide;
N-(3,5-dichloropyridin-4-yl)-2-[1-(4-fluorobenzyl)-6-hydroxyindol-3-yl]-2
oxoacetamide;
N-(3,5-dichloropyridin-4-yl)-5-hydroxy-1-(4-methoxybenzyl)indole-3-carboxam
ide.
The pharmaceutically acceptable salts are obtained in a customary
manner by neutralization of the bases with inorganic or organic
acids or by neutralization of the acids with inorganic or organic
bases. Possible inorganic acids are, for example, hydrochloric
acid, sulfuric acid, phosphoric acid or hydrobromic acid, organic
acids are, for example, carboxylic, sulfo or sulfonic acids such as
acetic acid, tartaric acid, lactic acid, propionic acid, glycolic
acid, malonic acid, maleic acid, fumaric acid, tannic acid,
succinic acid, alginic acid, benzoic acid, 2-phenoxybenzoic acid,
2-acetoxybenzoic acid, cinnamic acid, mandelic acid, citric acid,
malic acid, salicylic acid, 3-aminosalicylic acid, ascorbic acid,
embonic acid, nicotinic acid, isonicotinic acid, oxalic acid, amino
acids, methanesulfonic acid, ethanesulfonic acid,
2-hydroxyethanesulfonic acid, ethane-1,2-disulfonic acid,
benzenesulfonic acid, 4-methylbenzenesulfonic acid or
naphthalene-2-sulfonic acid. Possible inorganic bases are, for
example, sodium hydroxide solution, potassium hydroxide solution,
ammonia, and possible organic bases are amines, suitably tertiary
amines, such as trimethylamine, triethylamine, pyridine,
N,N-dimethylaniline, quinoline, isoquinoline, (-picoline,
(-picoline, (-picoline, quinaldine or pyrimidine.
In addition, pharmaceutically acceptable salts of the compound of
Formula (1) can be obtained by converting derivatives which have
tertiary amino groups into the corresponding quaternary ammonium
salts in a manner known per se by using quaternizing agents.
Possible quaternizing agents are, for example, alkyl halides such
as methyl iodide, ethyl bromide and n-propyl chloride, but also
arylalkyl halides such as benzyl chloride or 2-phenylethyl
bromide.
Furthermore, the invention of the compounds of Formula (1) which
contain an asymmetric carbon atom relates to the D form, the L form
and D,L mixtures and, in the case of a number of asymmetric carbon
atoms, the diastereomeric forms. Those compounds of Formula (1)
which contain asymmetric carbon atoms and as a rule are obtained as
racemates can be separated into the optically active isomers in a
manner known per se, for example using an optically active acid.
However, it is also possible to employ an optically active starting
substance from the start, a corresponding optically active or
diastereomeric compound then being obtained as the final
product.
The compounds of the present invention have therapeutically useful
pharmacological properties as inhibitors of the release of
TNF.alpha.. These disorders include, for example, arthritides
including arthritis and rheumatoid arthritis and other arthritic
disorders such as rheumatoid spondylitis and osteoarthritis.
Further possibilities of their application include the treatment of
patients suffering from sepsis, septic shock, gram-negative sepsis,
toxic shock syndrome, respiratory distress syndrome, asthma and
other chronic pulmonary disorders, bone resorption diseases or
transplant rejection reactions or other autoimmune disorders, such
as lupus erythematosus, multiple sclerosis, glomerulonephritis and
uveitis, insulin-dependent diabetes mellitus and chronic
demyelinization.
Moreover, the compounds of the present invention can also be
employed for the therapy of infections such as virus and parasite
infections, for example, for the therapy of malaria,
infection-related fever, infection-related myalgia, AIDS and
cachexia.
The compounds according to the invention are inhibitors of
phosphodiesterase 4 (PDE 4). Therefore, the compounds of Formula
(1) and their salts, and pharmaceutical preparations which contain
these compounds or their salts, can be used for the treatment of
disorders in which inhibition of phosphodiesterase 4 is
beneficial.
Thus the compounds according to the invention can be employed as
bronchodilators and for asthma prophylaxis. Compounds of Formula
(1) also inhibit of the accumulation and activity of eosinophils.
Accordingly, the compounds according to the invention can also be
employed in disorders in which eosinophils play a part. These
disorders include, for example, inflammatory airway disorders such
as bronchial asthma, allergic rhinitis, allergic conjunctivitis,
atopic dermatitis, eczema, allergic angiitis, inflammations
mediated by eosinophils such as eosinophilic fasciitis,
eosinophilic pneumonia and PIE syndrome (pulmonary infiltration
with eosinophilia), urticaria, ulcerative colitis, Crohn s disease
and proliferative skin disorders such as psoriasis or
keratosis.
According to the present invention the compounds of Formula (1) and
their salts can inhibit both the lipopolysaccharide (LPS)-induced
release of TNF.alpha. in human blood in vitro, and the LPS-induced
pulmonary neutrophilic infiltration in ferrets and domestic pigs in
vivo. All the pharmacologically important properties that were
found confirm that the compounds of Formula (1) and their salts as
well as pharmaceutical preparations which contain these compounds
or their salts can be used therapeutically for the treatment of
chronic obstructive pulmonary diseases.
The compounds of the invention also have neuroprotective properties
and can be used for the therapy of diseases in which
neuroprotection is beneficial. Such disorders are, for example,
senile dementia (Alzheimer's disease), loss of memory, Parkinson's
disease, depression, stroke and intermittent claudication.
Further applications of the compounds of the invention include the
prophylaxis and therapy of prostate diseases, such as, for example,
benign prostate hyperplasia, pollakiuria, nycturia, and for the
treatment of atony of the bladder and of colics caused by kidney
stones.
Finally, the compounds according to the invention can also be used
for the inhibition of the development of drug dependence on
repeated use of analgesics, such as, for example, morphine, and for
the reduction of the development of tolerance on repeated use of
these analgesics.
An efficective amount of the compounds according to the invention
or their salts is used for producing medicaments of the present
invention, along with conventional pharmaceutical auxiliaries,
carriers and additives.
The dose of the active compounds can vary depending on factors such
as the route of administration, age and weight of the patient,
nature and severity of the disorders to be treated and similar
factors. Therefore, any reference herein to a pharmacologically
effective amount of the compounds of the present invention refers
to the aforementioned factors.
The daily dose can be given as an individual dose to be
administered once or subdivided into two or more daily doses
suitably from about 0.001 mg to about 100 mg each.
Possible forms of administration include oral, parenteral,
intravenous, transdermal, topical, inhalational and intranasal
preparations. For administration, possible customary pharmaceutical
dosage forms include tablets, coated tablets, capsules, dispersible
powders, granules, aqueous solutions, aqueous or oily suspensions,
syrup, juices and drops.
Solid pharmaceutical forms can contain inert ingredients and
carriers, such as, for example, calcium carbonate, calcium
phosphate, sodium phosphate, lactose, starch, mannitol, alginates,
gelatin, guar gum, magnesium or aluminium stearates,
methylcellulose, talc, highly disperse salicylic acids, silicone
oil, high molecular weight fatty acids (such as stearic acid),
gelatin, agar-agar or vegetable or animal fats and oils, solid high
molecular weight polymers (such as polyethylene glycol);
preparations suitable for oral administration can, if desired,
contain additional flavorings and/or sweeteners.
Liquid pharmaceutical forms can be sterilized and/or optionally
contain auxiliaries such as preservatives, stabilizers, wetting
agents, penetrating agents, emulsifiers, spreading agents,
solubilizers, salts, sugars or sugar alcohols for regulation of the
osmotic pressure or for buffering, and/or viscosity regulators.
Additives of this type include, for example, tartrate and citrate
buffers, ethanol, complexing agents (such as
ethylenediaminetetraacetic acid and its nontoxic salts). For
regulation of the viscosity, possible high molecular weight
polymers are those such as, for example, liquid polyethylene oxide,
microcrystalline celluloses, carboxymethylcelluloses,
polyvinylpyrrolidones, dextrans or gelatin. Solid carriers include,
for example, starch, lactose, mannitol, methylcellulose, talc,
highly disperse salicylic acids, high molecular weight fatty acids
(such as stearic acid), gelatin, agar-agar, calcium phosphate,
magnesium stearate, animal and vegetable fats, solid high molecular
weight polymers such as polyethylene glycol.
Oily suspensions for parenteral or topical application can include
vegetable synthetic or semi-synthetic oils such as, for example,
liquid C.sub.8-22 fatty acid esters, for example palmitic, lauric,
tridecylic, margaric, stearic, arachidic, myristic, behenic,
pentadecanoic, linoleic, elaidic, brassidic, erucic or oleic acid,
which are esterified with mono- to C.sub.1-6 trihydric alcohols,
such as, for example, methanol, ethanol, propanol, butanol,
pentanol or their isomers, glycol or glycerol. Fatty acid esters of
this type are, for example, commercially available Miglyols,
isopropyl myristate, isopropyl palmitate, isopropyl stearate, PEG
6-capric acid, caprylic/capric acid esters of saturated fatty
alcohols, polyoxyethylene glycerol trioleates, ethyl oleate, waxy
fatty acid esters such as artificial duck preen gland fat,
isopropyl cocoate, oleyl oleate, decyl oleate, ethyl lactate,
dibutyl phthalate, diisopropyl adipate, polyol fatty acid esters
and others. Also suitable are silicone oils of differing
viscosities or fatty alcohols such as isotridecyl alcohol,
2-octyldodecanol, cetylstearyl alcohol or oleyl alcohol, fatty
acids such as, for example, oleic acid. Furthermore, vegetable oils
such as castor oil, almond oil, olive oil, sesame oil, cottonseed
oil, groundnut oil or soya bean oil can be used.
Possible solvents, gel-forming agents and solubilizers are water or
water-miscible solvents. Those suitable are, for example, alcohols
such as, for example, ethanol or isopropyl alcohol, benzyl alcohol,
2-octyldodecanol, polyethylene glycols, phthalates, adipates,
propylene glycol, glycerol, di- or tripropylene glycol, waxes,
methylcellosolve, cellosolve, esters, morpholines, dioxane,
dimethyl sulphoxide, dimethylformamide, tetrahydrofuran,
cyclohexanone etc.
Film-forming agents which can be used are cellulose ethers which
can dissolve or swell both in water and in organic solvents, such
as, for example, hydroxypropylmethylcellulose, methylcellulose,
ethylcellulose or soluble starches.
Mixed forms between gel- and film-forming agents are also possible.
Those used here are especially ionic macromolecules, such as, for
example, sodium carboxymethylcellulose, polyacrylic acid,
polymethacrylic acid and its salts, sodium amylopectin
semiglycolate, alginic acid or propylene glycol alginate as the
sodium salt, gum arabic, xanthan gum, guar gum or carrageenan.
Further formulation auxiliaries which can be employed include
glycerol, paraffin of differing viscosities, triethanolamine,
collagen, allantoin, novantisolic acid.
The use of surfactants, emulsifiers or wetting agents can also be
necessary for formulation, such as, for example, of Na lauryl
sulfate, fatty alcohol ether sulfates, di-Na
N-lauryl-(-iminodipropionate, polyethoxylated castor oil or
sorbitan monooleate, sorbitan monostearate, polysorbates (e.g.
Tween), cetyl alcohol, lecithin, glycerol monostearate,
polyoxyethylene stearate, alkylphenyl polyglycol ethers,
cetyltrimethylammonium chloride or mono-dialkyl polyglycol ether
orthophosphoric acid monoethanolamine salts.
Stabilizers such as montmorillonites or colloidal salicylic acids
for the stabilization of emulsions or for the prevention of the
breakdown of the active substances, such as antioxidants, for
example tocopherols or butylhydroxyanisole, or preservatives, such
as p-hydroxybenzoic acid esters, can likewise optionally be
required for the preparation of the desired formulations.
Preparations for parenteral administration can be present in
separate dose unit forms such as, for example, ampoules or vials.
Suitably, solutions of the active compound are used, most suitably
aqueous solutions and especially isotonic solutions, and also
suspensions. These injection forms can be made available as
finished preparations or prepared only directly before
administration by mixing the active compound, for example the
lyophilizate, if appropriate with further solid carriers, with the
desired solvent or suspending agent.
Intranasal preparations can be present as aqueous or oily solutions
or as aqueous or oily suspensions. They can also be present as
lyophilizates, which are prepared before administration using the
suitable solvent or suspending agent.
The production, dispensation and sealing of the preparations is
carried out under the conventional antimicrobial and aseptic
conditions.
The invention furthermore relates to processes for the preparation
of the compounds according to the invention.
According to the invention, the compounds of Formula (1) are
prepared by converting compounds of Formula (1), wherein R.sup.2 or
R.sup.3 or R.sup.2 and R.sup.3 are --O--R.sup.7, into the compounds
of the invention by removal of R.sup.7, wherein R.sup.7 is a
substituent suitable as a leaving group, such as, for example,
alkyl, cycloalkyl, arylalkyl, aryl, heteroaryl, acyl,
alkoxycarbonyl, aryloxycarbonyl, aminocarbonyl, N-substituted
aminocarbonyl, silyl or sulfonyl groups, and complexing agents,
such as, for example, compounds of boric acid, phosphoric acid and
covalently or coordinatively bonded metals, such as zinc, aluminium
or copper.
Particularly suitable reactions for the removal of R.sup.7 are
hydrolyses using suitable bases, such as, for example, sodium
hydroxide solution, potassium hydroxide solution or sodium
carbonate or potassium carbonate.
These hydrolyses are suitably used when R.sup.7 is an acyl,
alkoxycarbonyl, aryloxycarbonyl, aminocarbonyl, N-substituted
aminocarbonyl, silyl or sulfonyl residue, and a complexing agent,
such as, for example, compounds of boric acid, phosphoric acid and
coordinatively bonded metals, such as zinc, aluminium or copper.
Particularly suitable reactions for preparing the compounds of the
invention for the removal of R.sup.7 from the compounds in which
R.sup.7 is an alkyl, cycloalkyl, arylalkyl, aryl or heteroaryl
residue, are ether cleavages, for example by means of hydrobromic
acid, hydrochloric acid, hydriodic acid, and using activating Lewis
acids, such as, for example, AlCl3, BF3, BBr.sub.3 or LiCl, in each
case optionally in the presence of additional activators, such as,
for example, ethane-1,2-dithiol or benzyl mercaptan, and ether
cleavages by means of hydrogen, at elevated pressure or at normal
pressure, in the presence of a suitable catalyst, such as, for
example, a palladium or iridium catalyst.
According to the invention, the compounds of Formula (1) can also
be prepared by converting the substructure: ##STR3##
of compounds of Formula (1) by a reaction known per se into other
compounds of Formula (1). Particularly suitable conversion
reactions with compounds of Formula (1) are, for example, when A is
--(C.ident.O), reductions to result in A being --(CH--OH)-- or A
being --CH.sub.2 --, by reducing agents known per se, such as, for
example, sodium borohydride, or by hydrogenations, which can
optionally also be carried out stereoselectively.
Exemplary processes show below the preparation of compounds of
Formula (1) according to the invention from starting substances of
the type described, in which R.sup.7 is an alkyl, cycloalkyl,
arylalkyl, aryl or heteroaryl residue.
EXAMPLE 1
N-(3,5-Dichloropyridin-4-yl)-2-[1-(4-fluorobenzyl)-5-hydroxyindol-3-yl]-2-o
xoacetamide
1.4 g of
N-(3,5-dichloropyridin-4-yl)-2-[1-(4-fluorobenzyl)-5-methoxyindol-3-yl]-2-
oxoacetamide (3 mmol) is dissolved in 100 ml of dichloromethane.
The solution is heated to reflux and treated with a solution of 14
mmol of BBr.sub.3 in 15 ml of dichloromethane with stirring. The
reaction mixture is refluxed for 3 hours. After cooling, the
solution is intensively stirred for 3 hours at 20.degree. C. with
200 ml of an aqueous sodium hydrogencarbonate solution. The product
crystallizes out, it is isolated, dried at 60.degree. C. and
recrystallized from 80 ml of ethanol. Yield: 1.1 g (80% of theory)
Melting point: 213-214.degree. C.
EXAMPLE 2
N-(3,5-Dichloropyridin-4-yl)-2-[1-(4-fluorobenzyl)-5-hydroxyindol-3-yl)-]2-
oxoacetamide
5 g (38 mmol) anhydrous aluminium chloride is introduced into 50 ml
ethane-1,2,-dithiol. A solution of 4.7 g of
N-(3,5-dichloropyridin-4-yl)-2-[1-(4-fluorobenzyl)-5-methoxyindol-3-yl]-2-
oxoacetamide (10 mmol) in 50 ml of dichloromethane is added at
0.degree. C. The mixture is stirred at 0.degree. C. for 4 hours. 50
ml of 10% hydrochloric acid is added dropwise at from 0 to
10.degree. C. with stirring. The crystallizing product is isolated,
washed with water and dried at 20.degree. C. A pure product is
obtained by recrystallization from ethanol (180 ml). Yield: 3.1 g
(67% of theory) Melting point: 212-214.degree. C.
Exemplary preparative process as follows for compounds of Formula
(1) from starting substances of the type described, in which
R.sup.7 is an acyl, alkoxycarbonyl, aryloxycarbonyl, aminocarbonyl,
N-substituted aminocarbonyl, silyl or sulfonyl residue.
EXAMPLE 3
N-(3,5-Dichloropyridin-4-yl)-2-[1-(4-fluorobenzyl)-5-hydroxyindol-3-yl]-2-o
xoacetamide Na salt
5 g of
N-(3,5-dichloropyridin-4-yl)-2-[5-acetoxy-1-(4-fluorobenzyl)-indol-3-yl]2-
oxoacetamide (10 mmol) are stirred at 40.degree. C.-50.degree. C.
for 1 hour in 50 ml dilute sodium hydroxide solution. The solution
is neutralized with 10% hydrochloric acid while cooling with ice,
and is concentrated to dryness. The residue is dissolved in 80 mle
acetone and insoluble constituents are removed. The clear solution
is treated with a solution of 0.4 g NaOH in 3 ml of water and
stirred at 20.degree. C. for 2 hours. The crystallized product is
isolated, washed with acetone and dried at 60.degree. C. Yield:
2.44 g (51% of theory) Melting point: 265.degree. C.
An exemplary preparation process follows for compounds of Formula
(1) from other compounds of Formula (1).
EXAMPLE 4
N-(3,5-Dichloropyridin-4-yl)-2-[1-(4-fluorobenzyl)-5-hydroxyindol-3-yl]-2-h
ydroxyacetamide
1 g of
N-(3,5-dichloropyridin-4-yl)-2-[1-(4-fluorobenzyl)-5-hydroxyindol-3-yl]-2-
oxoacetamide (1; 2 mmol) are suspended in 75 ml methanol. After
addition of a solution of 0.2 g of sodium borohydride in 3 ml
dilute sodium hydroxide solution, the reaction mixture is stirred
at 20.degree. C. for 6 hours. After the solvent has been removed by
distillation, the residue is recrystallized from 40 ml ethanol.
Yield: 0.5 g (50% of theory) Melting point: 205-207.degree. C.
Numerous further compounds of Formula (1) can be prepared, as shown
in the Examples and also in the further examples, all summarized in
the next table.
Melting point Ex. R.sup.1 R.sup.2 R.sup.3 R.sup.4 R.sup.5 A B D E
[.degree. C.] 1 4-Fluorobenzyl --OH --H --H 3,5-Dichloro-4-pyridyl
--(C.dbd.O)-- C O --(N--H)-- 215 2 4-Fluorobenzyl --O.sup.- --H --H
3,5-Dichloro-4-pyridyl --(C.dbd.O)-- C O --(N--H)-- 265 Na.sup.+ 3
4-Fluorobenzyl --OH --H --H 3,5-Dichloro-4-pyridyl --(CHOH)-- C O
--(N--H)-- 205-207 4 2,6-Diflurorbenzyl --OH --H --H 4-Pyridyl
--(C.dbd.O)-- C O --(N--H)-- 327-329 5 2,6-Diflurorbenzyl --OH --H
--H 3,5-Dichloro-4-pyridyl --(C.dbd.O)-- C O --(N--H)-- 266-268 6
3-Nitrobenzyl --O.sup.- --H --H 3,5-Dichloro-4-pyridyl
--(C.dbd.O)-- C O --(N--H)-- 235-238 Na.sup.+ dec. 7 n-Propyl --OH
--H --H 3,5-Dichloro-4-pyridyl --(C.dbd.O)-- C O --(N--H)-- 280-282
8 Isopropyl --OH --H --H 3,5-Dichloro-4-pyridyl --(C.dbd.O)-- C O
--(N--H)-- 245-247 9 Cyclopentylmethyl --OH --H --H
3,5-Dichloro-4-pyridyl --(C.dbd.O)-- C O --(N--H)-- 246-248 10
4-Fluorobenzyl --OH --H --H 2,6-Dichlorophenyl --(C.dbd.O)-- C O
--(N--H)-- 216-218 11 4-Fluorobenzyl --OH --H --H 2,6-Dichloro-4-
--(C.dbd.O)-- C O --(N--H)-- 199-201 trifluoromethylphenyl 12
4-Fluorobenzyl --OH --H --H 2,6-Dichloro-4- --(C.dbd.O)-- C O
--(N--H)-- 176-178 trifluoromethoxyphenyl 13 4-Fluorobenzyl --H
--OH --H 3,5-Dichloro-4-pyridyl --(C.dbd.O)-- C O --(N--H)--
212-213 14 4-Methoxybenzyl --OH --H --H 3,5-Dichloro-4-pyridyl -- C
O --(N--H)-- 239-241
The compounds according to the invention are strong inhibitors of
phosphodiesterase 4 and TNF.alpha. release. Their therapeutic
potential is confirmed in vivo, for example, by the inhibition of
the asthmatic late-phase reaction (eosinophilia) in guinea-pigs and
by the influencing of the allergen-induced vascular permeability in
actively-sensitized brown Norway rats.
The PDE 4 inhibiting activity is determined in enzyme preparations
of human polymorphonuclear lymphocytes (PMNLs), the PDE 2, 3 and 5
activity with PDE from human platelets. Human blood was
anticoagulated with citrate. The thrombocyte-rich plasma in the
supernatant is separated from the erythrocytes and leucocytes by
centrifugation at 700.times.g for 20 minutes at RT. The platelets
are lysed by ultrasound and employed in the PDE 3 and PDE 5 assay.
For the determination of the PDE 2 activity, the cytosolic platelet
fraction is purified on an anion exchange column by means of NaCl
gradients and the PDE 2 peak is recovered for the assay. The PMNLs
for the PDE 4 determination are isolated by a following dextran
sedimentation and subsequent gradient centrifugation using
Ficoll-Paque. After a second washing of the cells, the erythrocytes
still contained are lysed in the course of 6 minutes at 4.degree.
C. by the addition of 10 ml of hypotonic buffer (155 mM NH.sub.4
Cl, 10 mM NaHCO.sub.3, 0.1 mM EDTA, pH 7.4). The still intact PMNLs
are washed with PBS a further two times and lysed by means of
ultrasound. The supernatant of a one-hour centrifugation at
4.degree. C. at 48,000.times.g contains the cytosolic fraction of
the PDE 4 and is employed for the PDE 4 measurements.
The phosphodiesterase activity is determined with some
modifications according to the method described by Thompson et al.
(Thompson, W. J.; Appleman, M. M., Assay of cyclic nucleotide
phosphodiesterase and resolution of multiple molecular forms of the
enzyme, Adv. Cycl. Nucl. Res. 1979, 10, of multiple molecular forms
of the enzyme, Adv. Cycl. Nucl. Res. 1979, 10, 69-92).
The reaction mixtures contain 50 mM tris HCl (pH 7.4), 5 mM
MgCl.sub.2, the inhibitors in variable concentrations, the
corresponding enzyme preparation and also the further components
necessary for the detection of the individual isoenzymes (see
below). The reaction is started by the addition of the substrate
0.5 .mu.M [.sup.3 H]-cAMP or [.sup.3 H]-cGMP (about 6000 CPM/test).
The final volume is 100 ml. Test substances are prepared as stock
solutions in DMSO. The DMSO concentration in the reaction mixture
is 1% v/v. At this DMSO concentration, the PDE activity is not
affected. After the start of the reaction by means of substrate
addition, the samples are incubated at 37.degree. C. for 30
minutes. The reaction is stopped by heating the test tubes for 2
minutes at 110.degree. C. The samples remain in the ice for a
further 10 minutes. After the addition of 30 .mu.l of
5-nucleotidase (1 mg/ml, of a snake venom suspension from Crotalus
adamanteus) incubation is carried out for 10 minutes at 37.degree.
C. The samples are stopped on ice, 400 .mu.l each of a mixture of
Dowex-water-ethanol (1+1+1) are added, and the samples are well
mixed and again incubated on ice for 15 minutes. The reaction
vessels are centrifuged at 3000.times.g for 20 minutes. 200 .mu.l
aliquots of the supernatant are transferred directly to
scintillation vessels. After the addition of 3 ml of scintillator,
the samples are measured in a beta counter.
[.sup.3 H]-cAMP is used as a substrate for the determination of the
PDE 4, 3 and 2 activity, [.sup.3 H]-cGMP for the determination of
the PDE 5 activity. The non-specific enzyme activities in each case
are determined in the presence of 100 .mu.M rolipram in the case of
PDE 4 and in the presence of 100 .mu.M IBMX in the determination of
PDE 3 and 5 and subtracted from the test values. The incubation
batches of the PDE 3 assay contain 10 .mu.M rolipram in order to
inhibit possible contamination by the PDE 4. The PDE 2 is tested
using an SPA assay from Amersham. The assay is carried out in the
presence of the activator of PDE 2 (5 .mu.M cGMP).
IC.sub.50 values in the range from 10.sup.-9 to 10.sup.-5 M were
calculated for the compounds according to the invention in relation
to the inhibition of phosphodiesterase 4. The selectivity to the
PDE types 2, 3 and 5 is factor 100 to 10,000.
For the determination of the inhibition of TNF.alpha. release from
cells of nasal polyps, the experimental arrangement essentially
corresponds to the method described by Campbell, A. M. and Bousquet
J (Anti-allergic activity of H.sub.1 -blockers, Int. Arch. Allergy
Immunol., 1993, 101, 308-310). The starting material is nasal
polyps (obtained from operation) of patients who have been
subjected to surgical treatment.
The tissue is washed with RPMI 1640 and then broken down at
37.degree. C. for 2 hours using protease (2.0 mg/ml), collagenase
(1.5 mg/ml), hyaluronidase (0.75 mg/ml) and DNAse (0.05 mg/ml) (1 g
of tissue to 4 ml of RPMI 1640 with enzymes). The cells obtained, a
mixture of epithelial cells, monocytes, macrophages, lymphocytes,
fibroblasts and granulocytes, are filtered and washed by repeated
centrifugation in nutrient solution, passively sensitized by
addition of human IgE and the cell suspension is adjusted to a
concentration of 2 million cells/ml in RPMI 1640 (supplemented with
antibiotics, 10% foetal calf serum, 2 mM glutamine and 25 mM
Hepes). This suspension is distributed in 6-well cell culture
plates (1 ml/well). The cells are preincubated for 30 min with the
test substances in various final concentrations and then stimulated
to TNF.alpha. release by addition of anti-IgE (7.2 .mu.g/ml). The
maximum release into the nutrient medium takes place after about 18
hours. In this period, the cells are incubated at 37.degree. C. and
5% CO.sub.2. The supernatant nutrient medium is recovered by
centrifugation (5 min, 4000 rpm) and stored at -70.degree. C. until
cytokine determination. The determination of TNF.alpha. in the
supernatant is carried out using so-called sandwich ELISAs (basic
material Pharmingen), in which concentrations of the cytokine in
the range from 30-1000 pg/ml can be detected. Cells not stimulated
with anti-IgE barely produce TNF.alpha., stimulated cells, however,
secrete large amounts of TNF.alpha., which can be decreased in a
dose-dependant manner, for example, by PDE 4 inhibitors. The
IC.sub.50 (concentration at 50% inhibition) is calculated from the
percentage inhibition (TNF.alpha. release of the cells stimulated
with anti-IgE=100%) of the tested substances at various
concentrations.
For the compounds according to the present invention, IC.sub.50
values in the range of 10.sup.-7 to 10.sup.-5 M were
determined.
The inhibition of the pulmonary eosinophil infiltration by the
substances is investigated in an in vivo test of the inhibition of
the late-phase eosinophilia 24 hours after inhalational ovalbumin
challenge of actively sensitized guinea-pigs on male Dunkin-Hartley
guinea-pigs (200-250 g) actively sensitized against ovalbumin
(OVA). The sensitization is carried out by means of two
intraperitoneal injections of a suspension of 20 .mu.g of OVA
together with 20 mg of aluminium hydroxide as an adjuvant in 0.5 ml
of physiological saline solution per animal on two successive days.
14 days after the second injection, the animals are pretreated with
mepyramine maleate (10 mg/kg i.p.) in order to protect them from
anaphylactic death. 30 minutes later, the animals are exposed for
30 sec in a plastic box to an OVA aerosol (0.5 mg/ml) which is
generated by a nebulizer driven with compressed air (19.6 kPa)
(allergen challenge). Control animals are nebulized with
physiological saline solution. 24 hours after the challenge, the
animals are anaesthetized with an overdose of ethylurethane (1.5
g/kg of body weight i.p.) and a bronchoalveolar lavage (BAL) is
carried out using 2.times.5 ml of physiological saline solution.
The BAL fluid is collected, centrifuged at 300 rpm for 10 min and
the cell pellet is then resuspended in 1 ml of physiological saline
solution. The eosinophils in the BAL are counted using an automatic
cell differentiation apparatus (Bayer Diagnostics Technicon H1). 2
control groups (nebulization with physiological saline solution and
nebulization with OVA solution) are included in each test.
The percentage inhibition of eosinophilia of the test group treated
with substance is calculated according to the formula: ##EQU1##
wherein
A is eosinophils in the control group with OVA challenge and
vehicle
B is eosinophils in the group with OVA challenge treated with
substance
C is eosinophils in the control group with 0.9% strength NaCl
challenge and vehicle
The test substances are administered intraperitoneally or orally as
a suspension in 10% polyethylene glycol 300 and 0.5% strength
5-hydroxy-ethylcellulose 2 hours before the allergen challenge. The
control groups are treated with the vehicle according to the form
of administration of the test substance.
The compounds according to the invention were found to inhibit
late-phase eosinophilia by 30% to 80% after intraperitoneal
administration of 10 mg/kg and by 40% to 70% after oral
administration of 30 mg/kg. The compounds according to the
invention are thus particularly suitable for the production of
drugs for the treatment of disorders which are connected with the
action of eosinophils.
The effect of allergen-induced vascular permeability was determined
on actively sensitized male brown Norway rats. Male brown Norway
rats weighing 280-300 g are actively sensitized on 2 successive
days by intraperitoneal injection of a suspension of 1 mg of
ovalbumin together with 100 mg of aluminium hydroxide in 1
ml/animal. Three weeks after sensitization, the rats are
anaesthetized with sodium thiopental and fixed in the supine
position. A polyethylene catheter was advanced into the trachea in
a backward direction as far as the internal opening of the choanas
for perfusion of the nasal cavity, so that it was possible for the
solution to trickle out through the nasal cavities. A short
tracheal catheter was tied into the trachea in an orthograde manner
to make respiration possible. Phosphate-buffered saline solution
(PBS) was continuously pumped for perfusion through the nasal
cavity (0.5 ml/min) using a roller pump and collected by means of a
fraction collector. Evans Blue was used as a plasma marker and
injected intravenously (1 ml/animal each of a 1% strength solution
in PBS) through a catheter in the jugular vein.
Substance administration was carried out topically. During
administration, the test substance was added to the perfusion
medium (PBS). The nasal mucous membrane was perfused for 30 min
with PDE 4 inhibitor-containing solution. Evans Blue was then
injected immediately before the start of the perfusion with
ovalbumin-containing solution (challenge). After the start of the
ovalbumin challenge (10 mg/ml of ovalbumin dissolved in PBS) 15 min
fractions were collected every 15 min in the fraction collector
over a period of 60 min. The Evans Blue concentration in the
perfusates was measured with a Digiscan photometer at a wavelength
of 620 nm. The blank values were automatically subtracted in the
course of this. The course of action over 60 min was calculated
using an AUC program. The substance action of the preparation group
was calculated against vehicle controls in %.
IC.sub.50 values in the range from 10.sup.-8 to 10.sup.-5 M were
determined for the compounds of the present invention.
The utility of the compounds according to the invention of Formula
(I) for the therapy of chronic obstructive pulmonary diseases is
confirmed by the inhibition of LPS-induced TNF.alpha. release in
human blood and by the inhibition of LPS-induced pulmonary
neutrophil infiltration in ferrets and domestic pigs, all good
animal models.
The stimulation of isolated leucocytes to cytokine release can take
place in various ways. Lipopolysaccharides (LPSs) are a stimulus
suitable for the investigation of TNF.alpha. release. LPS is a
constituent of the bacterial cell walls and is released by killing
the bacteria (antibiotics or immune system). LPS particularly
stimulates the activity of the phagocytizing leucocytes (tissue
macrophages, granulocytes, monocytes) and causes the infiltration
of leucocytes from the blood stream into the affected tissue. A
cytokine important for these mechanisms is TNF.alpha., which is
secreted in large amounts by the affected cells (the monocytes and
macrophages are the main source) and initiates and maintains
inflammation alongside other mediators.
For the investigation of the effect on LPS-induced TNF.alpha.
release, human blood was obtained from various donors (inhibition
of coagulation by means of citrate) and diluted 1:5 with RPMI 1640
cell culture medium. The test substances were added to the samples
in various concentrations before the LPS challenge. The stimulation
of the leucocytes was carried out 30 min later using
lipopolysaccharides (LPS) from Salmonella abortus equi in a final
concentration of 10 .mu.g/ml. After incubation of the test batches
for 24 hours at 37.degree. C. and under 5% CO.sub.2 in an
incubator, the diluted blood was centrifuged and the TNF.alpha.
concentration in the cell-free supernatant was measured by means of
ELISA.
IC.sub.50 values in the range from 10.sup.-7 to 10.sup.-5 M were
determined for the compounds according to the invention. An
IC.sub.50 value of 0.8 .mu.mol/l, for example, was determined for
the compound as in Example 1. In comparison with this, an IC.sub.50
value of 7.0 .mu.mol/l was determined with the reference standard
SB 207499.
The inhibition of lipopolysaccharide (LSP)-induced pulmonary
neutrophil infiltration by the substance is investigated in an in
vivo test on male ferrets (0.6-2 kg). The experimental animals are
anaesthetized with pentobarbital sodium (40 mg/kg of body weight
i.p.), placed individually into a closed nebulization box of 5 l
capacity and exposed to an ultrasonically nebulized aerosol of
0.01% strength LPS (lipopolysaccharide) solution (additionally 0.1%
hydroxylamine in PBS) for 10 minutes. The aerosol is generated by a
nebulizer driven with compressed air (0.2 Mpa). Control animals are
treated with an aerosol of physiological saline solution. The
animals are observed during the entire process and removed from the
nebulization box after admission of fresh air. On inhalation,
nebulized LPS immediately induces inflammation of the airways,
which is characterized by a massive infiltration of neutrophilic
granulocytes into the lungs of the experimental animals. The
neutrophilia achieves its maximum 4 to 6 hours after LPS exposure.
In order to be able to measure the number of infiltrated
neutrophilic granulocytes, the animals are anaesthetized with an
overdose of ethylurethane (1.5 g/kg of body weight i.p.) 6 hours
after LPS provocation and a bronchoalveolar lavage (BAL) is carried
out using 2.times.10 ml of physiological saline solution. The
number of cells in the pooled original BAL fluid (100 .mu.l) are
determined using the automatic cell-counting apparatus sold by
Bayer Diagnostic under the trade designation Technicon H1E and the
different leucocytes per .mu.l are differentiated. In each test, 2
control groups (nebulization with physiological saline solution or
with LPS solution) are included. Substances having
anti-inflammatory activity, particularly those which affect
TNF.alpha. release or the function of the neutrophilic
granulocytes, inhibit the infiltration of leucocytes. The
inhibition of infiltration is determined by the comparison of the
number of infiltrated neutrophils in untreated experimental animals
(with and without LPS provocation).
ID.sub.50 values in the range from 1 to 20 mg/kg i.p. were
determined for the compounds according to the invention. The
compound of Example 1 was administered in doses of 1, 3 and 10
mg/kg i.p. 2 hours before LPS provocation to up to 3 experimental
animals per dose. The neutrophilia in the BAL was inhibited in a
dose-dependent manner (18%, 64% and 78%). The ID.sub.50 is 2.4
mg/kg i.p. The administration of the selected PDE 4 inhibitor
RPR-73401 (reference substance) caused an inhibition of
neutrophilia of 49% in the dose 1 mg/kg i.p.
For intrapulmonary administration, the trachea of the animals is
opened under anaesthesia with 40 mg/kg i.p. of pentobarbital
sodium, 3% strength, 1.3 ml/kg, a 7 cm-long PVC catheter is tied in
and the test substances are administered intrapulmonarily in powder
form (mixed with lactose to 20 mg/kg) by means of a syringe 2 hours
before LPS provocation. The intrapulmonary administration of
Example 1 in doses of 1, 3 and 10 mg/kg inhibits LPS-induced
neutrophilia in a dose-dependent manner (43%, 65% and 100%). The
ID.sub.50 is 1.65 mg/kg i.palm.
Pulmonary neutrophilia can be induced with LPS in domestic pigs in
a manner similar to that in the ferret. The animals are
anaesthetized with pentobarbital 10 mg/kg i.v., and intubated.
Using a bronchoscope, a partial bronchoalveolar lavage is carried
out in order to determine the proportion of neutrophilic
granulocytes under physiological conditions. The test substance is
then administered and the animals inhale an ultrasonically
nebulized aerosol of 0.03% strength LPS (lipopolysaccharide)
solution (additionally 0.1% hydroxylamine in PBS) through the
tracheal tube for 20 min. The inhaled LPS induces a reactive
inflammation of the airways and neutrophilic granulocytes
infiltrate on a huge scale. The neutrophilia achieves its maximum 4
to 6 hours after LPS exposure. After 6 hours, the bronchoalveolar
lavage is repeated and the increase in the neutrophil count is
determined arithmetically.
Among animal species, the pig is particularly suitable for these
investigations, since is has large anatomical and physiological
similarities to man. For the compounds according to the invention,
inhibitions of LPS-induced neutrophilia of 20% to 65% were
determined on intrapulmonary administration of 10 mg/animal.
The intrapulmonary administration of the compound of Example 1 in
the dose 10 mg/animal (about 0.75 mg/kg) inhibited LPS-induced
pulmonary neutrophilia by 51%.
* * * * *