Title of Invention

SUBSTITUTED QUINOLINES AND THEIR USE AS MYCOBACTERIAL INHIBITORS

Abstract The present invention relates to a compound of formula the pharmaceutically acceptable acid or base addition salts thereof and process for preparing the same.
Full Text SUBSTITUTED QUINOLINES AND THEIR USE AS MYCOBACTERIAL INHIBITORS
The present invention relates to novel substituted quinoline derivatives useful for the
treatment of mycobaclerial diseases, particularly those diseases caused by pathogenic
mycobacteria such as Mycobacterium (M.) tuberculosis, M. bovis, M. avium andM.
martnum.
BACKGROUND OF THE INVENTION
Mycobacterium tuberculosis is the causative agent of tuberculosis (TB), a serious and
potentially fatal infection with a world-wide distribution. Estimates from the World
Health Organization indicate that more than 8 million people contract TB each year,
and 2 million people die from tuberculosis yearly. In the last decade, TB cases have
grown 20% worldwide with the highest burden hi the most impoverished communities.
If these trends continue, TB incidence will increase by 41% in the next twenty years.
Fifty years since the introduction of an effective chemotherapy, TB remains after
ADDS, the leading infectious cause of adult mortality in the world. Complicating the TB
epidemic is the rising tide of multi-drug- resistant strains, and the deadly symbiosis
with HTV. People who are HIV-positive and infected with TB are 30 times more likely
to develop active TB than people who are HIV-negative and TB is responsible for the
death of one out of every three people with HIV/AIDS worldwide.
Existing approaches to treatment of tuberculosis all involve the combination of multiple
agents. For example, the regimen recommended by the U.S. Public Health Service is a
combination of isoniazid, rifampicin and pyrazinarnide for two months, followed by
isoniazid and rifampicin alone for a further four months. These drugs are continued for
a further seven months hi patients infected with HTV. For patients infected with multidrug
resistant strains of M tuberculosis, agents such as ethambutol, streptomycin,
kanamycin, amileacin, capreomycin, ethionamide, cycloserine, ciprofoxacin and
ofloxacin are added to the combination therapies. There exists no single agent that is
effective in the clinical treatment of tuberculosis, nor any combination of agents that
offers the possibility of therapy of less than six months duration.
There is a high medical need for new drugs that improve current treatment by enabling
regimens that facilitate patient and provider compliance. Shorter regimens and those
that require less supervision are the best way to achieve this. Most of the benefit from
treatment comes in the first 2 months, during the intensive, or bactericidal, phase when
four drugs are given together; the bacterial burden is greatly reduced, and patients
become noninfectious. The 4- to 6-month continuation, or sterilizing, phase is required
to eliminate persisting bacilli and to minimize the risk of relapse. A potent sterilizing
drug that shortens treatment to 2 months or less would be extremely beneficial. Drags
that facilitate compliance by requiring less intensive supervision also are needed.
Obviously, a compound that reduces both the total length of treatment and the
frequency of drug administration would provide the greatest benefit.
Complicating the TB epidemic is the increasing incidence of multi-drug- resistant
strains or MDR-TB. Up to four percent of all cases worldwide are considered MDR-TB
- those resistant to the most effective drugs of the four-drug standard, isoniazid and
rifampin. MDR-TB is lethal when untreated and can not be adequately treated through
the standard therapy, so treatment requires up to 2 years of "second-line" drugs. These
drugs are often toxic, expensive and marginally effective. In the absence of an effective
therapy, infectious MDR-TB patients continue to spread the disease, producing new
infections with MDR-TB strains. There is a high medical need for a new drug with a
new mechanism of action, which is likely to demonstrate activity against MDR strains.
The term "drug resistant" as used hereinbefore or hereinafter is a term well understood
by the person skilled in microbiology. A drug resistant Mycobacterium is a
Mycobacterium which is no longer susceptible to at least one previously effective drug;
which has developed the ability to withstand antibiotic attack by at least one previously
effective drug. A drug resistant strain may relay that ability to withstand to its progeny.
Said resistance may be due to random genetic mutations in the bacterial cell that alters
its sensitivity to a single drug or to different drugs.
MDR tuberculosis is a specific form of drug resistant tuberculosis due to a bacterium
resistant to at least isoniazid and rifampicin (with or without resistance to other drugs),
which are at present the two most powerful anti-TB drugs.
The purpose of the present invention is to provide novel compounds, in particular
substituted quinoline derivatives, having the property of inhibiting growth of
Mycobacteria including drug resistant or multi drug resistant Mycobacteria, and
therefore useful for the treatment of mycobacterial diseases, particularly those diseases
caused by pathogenic mycobacteria such as Mycobacterium tuberculosis, M. bovis,
M. aviwn, M. smegmatis andM. marinum.
Substituted quinolines were already disclosed in US 5,965,572 (The United States of
America) for treating antibiotic resistant infections and in WO 00/34265 to inhibit the
growth of bacterial microorganisms. None of these publications disclose the
substituted quinoline derivatives according to our invention.
SUMMARY OF THE INVENTION
The present invention relates to novel substituted quinoline derivatives according to
Formula (la) and (Ib)
the pharmaceutically acceptable acid or base addition salts thereof, the quaternary
amines thereof, the stereochemically isomeric forms thereof, the tautomeric forms
thereof and the JV-oxide forms thereof, wherein:
R1 is hydrogen, halo, haloalkyl, cyano, hydroxy, Ar, Het, alkyl, alkyloxy,
alkylthio, alkyloxyalkyl, alkylthioalkyl, Ar-alkyl or di(Ar)alkyl;
p is an integer equal to 1,2, 3 or 4;
R2 is hydrogen, hydroxy, thio, alkyloxy, alkyloxyalkyloxy, alkylthio, mono
or di(alkyl)amino or a radical of formula wherein Y is
O,S,NHorN-alkyl;
R3 is alkyl, Ar, Ar-alkyl, Het or Het-alkyl;
R4 is hydrogen, alkyl or benzyl;
R5 is hydrogen, halo, haloalkyl, hydroxy, Ar, alkyl, alkyloxy, alkyMhio,
alkyloxyalkyl, alkylthioalkyl, Ar-alkyl or di(Ar)alkyI; or
two vicinal R5 radicals may be taken together to form together with the phenyl ring to
which they are attached a naphthyl;
r is an integer equal to 1,2,3,4 or 5; and
R6 is hydrogen, alkyl, Ar or Het;
R7 is hydrogen or alkyl;
R8 is oxo; or
R7 and R8 together form the radical -CH=CH-N=;
Z isCH2orC(=O);
alkyl is a straight or branched saturated hydrocarbon radical having from 1 to 6
carbon atoms; or is a cyclic saturated hydrocarbon radical having from 3 to 6
carbon atoms; or is a a cyclic saturated hydrocarbon radical having from 3 to 6
carbon atoms attached to a straight or branched saturated hydrocarbon radical
having from 1 to 6 carbon atoms ; wherein each carbon atom can be optionally
substituted with halo, hydroxy, alkyloxy or oxo;
Ar is a homocycle selected from the group of phenyl, naphthyl, acenaphthyl,
tetrahydronaphthyl, each optionally substituted with 1,2 or 3 substituents, each
substituent independently selected from the group of hydroxy, halo, cyano,
nitro, amino, mono- or dialkylamino, alkyl, haloalkyl, alkyloxy, haloalkyloxy,
carboxyl, alkyloxycarbonyl, aminocarbonyl, morpholinyl and mono- or
dialkylaminocarbonyl;
Het is a monocyclic heterocycle selected from the group of N-phenoxypiperidinyl,
pyrrolyl, pyrazolyl, imidazolyl, furanyl, thienyl, oxazolyl, isoxazolyl, thiazolyl,
isothiazolyl, pyridinyl, pyrimidinyl, pyrazinyl and pyridazinyl; or a bicyclic
heterocycle selected from the group of quinolinyl, quinoxalinyl, indolyl,
benzunidazolyl, benzoxazolyl, benzdsoxazolyl, benzofhiazolyl, benzisothiazolyl,
benzofuranyl, benzothienyl, 2,3-dihydrobenzotl,4]dioxinyl or
benzo[l,3]dioxolyl; each monocyclic and bicyclic heterocycle may optionally
be substituted on a carbon atom with 1,2 or 3 substituents selected from the
group of halo, hydroxy, alkyl or alkyloxy;
halo is a substituent selected ftom the group of fluoro, chloro, bromo and iodo and
haloalkyl is a straight or branched saturated hydrocarbon radical having from 1 to
6 carbon atoms or a cyclic saturated hydrocarbon radical having from 3
to 6 carbon atoms, wherein one or more carbon atoms are substituted
with one or more halo-atoms.
The compounds according to Formula (la) and (Ib) are interrelated in that e.g. a
compound according to Formula (Ib), with R8 equal to oxo is the tautomeric equivalent
of a compound according to Formula (la) with R2 equal to hydroxy (keto-enol
tautomerism).
DETAILED DESCRIPTION
In the framework of this application, alkyl is a straight or branched saturated
hydrocarbon radical having from 1 to 6 carbon atoms ; or is a cyclic saturated
hydrocarbon radical having from 3 to 6 carbon atoms ; or is a a cyclic saturated
hydrocarbon radical having from 3 to 6 carbon atoms attached to a straight or branched
saturated hydrocarbon radical having from 1 to 6 carbon atoms ; wherein each carbon
atom can be optionally substituted with halo, hydroxy, alkyloxy or oxo.
Preferably, alkyl is methyl, ethyl or cyclohexylmethyl.
In the framework of this application, Ar is a homocycle selected from the group of
phenyl, naphthyl, acenaphthyl, tetrahydronaphthyl, each optionally substituted with 1,2
or 3 substituents, each substituent independently selected from the group of hydroxy,
halo, cyano, nitro, amino, mono- or dialkylamino, alkyl, haloalkyl, alkyloxy,
haloalkyloxy, carboxyl, alfcyloxycarbonyl, aminocarbonyl, morpholinyl and mono- or
dialkylaminocarbonyl. Preferably, Ar is naphthyl or phenyl, each optionally
substituted with 1 or 2 halo substituents.
In the framework of this application, Het is a monocyclic heterocycle selected from the
group of N-phenoxypiperidinyl, pyrrolyl, pyrazolyl, imidazolyl, furanyl, thienyl,
oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyridinyl, pyrimidinyl, pyrazinyl and
pyridazinyl; or a bicyclic heterocycle selected from the group of quinolinyl,
quinoxalinyl, indolyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzotbiazolyl,
benzisothiazolyl, benzofuranyl, benzothienyl, 2,3-dihydrobenzo[l,4]dioxinyl or
benzo[l,3]dioxolyl; each monocyclic and bicyclic heterocycle may optionally be
substituted on a carbon atom with 1,2 or 3 substituents selected firom the group of halo,
hydroxy, alkyl or alkyloxy. Preferably, Het is thienyl or furanyl or pyridyl, most
preferably Het is fiiranyl.
In the framework of this application, halo is a substituent selected from the group of
fluoro, chloro, bromo and iodo and haloalkyl is a straight or branched saturated
hydrocarbon radical having from 1 to 6 carbon atoms or a cyclic saturated hydrocarbon
radical having from 3 to 6 carbon atoms, wherein one or more carbon atoms are
substituted with one or more halo-atoms. Preferably, halo is bromo, fluoro or chloro
and preferably, haloalkyl is trifluoromethyl.
Whenever used hereinafter, the term "compounds of formula (la) or (Ib)" is meant to
also include their JV-oxide forms, their salts, their quaternary amines, their tautomeric
forms and their stereochemically isomeric forms. Of special interest are those
compounds of formula (la) or (Ib) which are stereochemically pure.
An interesting embodiment of the present invention relates to those compounds of
formula (la) or (Ib), the pharmaceutically acceptable acid or base addition salts thereof,
the stereochemically isomeric forms thereof, the tautomeric forms thereof and the
JV-oxide forms thereof, wherein
Z is CH2;
R1 is hydrogen, halo, haloalkyl, cyano, hydroxy, Ar, Het, alkyl, alkyloxy,
alkylthio., alkyloxyalkyl, alkylthioalkyl, Ar-alkyl or di(Ar)alkyl;
p is an integer equal to 1,2, 3 or 4;
R2 is hydrogen, hydroxy, thio, alkyloxy., alkyloxyalkyloxy, alkylthio, mono
or di(alkyl)amino or a radical of formula ^^ wherein Y is
O, S,NHorN-alkyl;
R3 is alkyl, Ar, Ar-alkyl, Het or Het-alkyl;
R4 is hydrogen, alkyl or benzyl;
R5 is hydrogen, halo, haloalkyl, hydroxy, Ar, alkyl, alkyloxy, alkylthio,
alkyloxyalkyl, alkylthioalkyl, Ar-alkyl or di(Ar)alkyl; or
two vicinal R5 radicals may be taken together to form together with the phenyl ring to
which they are attached a naphthyl;
r is an integer equal to 1, 2, 3,4 or 5 ; and
R6 is hydrogen, alkyl, Ar or Het;
R7 is hydrogen or alkyl;
R8 is oxo; or
R7 and R8 together form the radical -CH=CH-N=;
alkyl is a straight or branched saturated hydrocarbon radical having from 1 to 6
carbon atoms; or is a cyclic saturated hydrocarbon radical having from 3 to 6
carbon atoms; or is a a cyclic saturated hydrocarbon radical having from 3 to 6
carbon atoms attached to a straight or branched saturated hydrocarbon radical
having from 1 to 6 carbon atoms ; wherein each carbon atom can be optionally
substituted with halo, hydroxy, alkyloxy or oxo;
Ar is a homocycle selected from the group of phenyl, naphthyl, acenaphthyl,
tetrahydronaphthyl, each optionally substituted with 1,2 or 3 substituents, each
substituent independently selected from the group of hydroxy, halo, cyano,
nitro, amino, mono- or dialkylamino, alkyl, haloalkyl, alkyloxy, haloalkyloxy,
carboxyl, alkyloxycarbonyl, aminocarbonyl, morpholinyl and mono- or
dialkylaminocarbonyl;
Het is a monocyclic heterocycle selected from the group of N-phenoxypiperidinyl,
pyrrolyl, pyrazolyl, imidazolyl, furanyl, thienyl, oxazolyl, isoxazolyl, thiazolyl,
isothiazolyl, pyridinyl, pyrimidinyl, pyrazinyl andpyridazinyl; or a bicyclic
heterocycle selected from the group of quinolinyl, quinoxalinyl, indolyl,
benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzothiazolyl, benzisotbiazolyl,
benzofuranyl, benzothienyl, 2,3-dihydrobenzo[l,4]dioxinyl or
benzo[l,3]dioxolyl; each monocyclic and bicyclic heterocycle may optionally
be substituted on a carbon atom with 1,2 or 3 substituents selected from the
group of halo, hydroxy, alkyl or alkyloxy;
halo is a substituent selected from the group of fluoro, chloro, bromo and iodo and
haloalkyl is a straight or branched saturated hydrocarbon radical having from 1 to
6 carbon atoms or a cyclic saturated hydrocarbon radical having from 3
to 6 carbon atoms, wherein one or more carbonatoms are substituted
with one or more halo-atoms.
Preferably, R5 is hydrogen, halo, haloalkyl, hydroxy, Ar, alkyl, alkyloxy, alkylfhio,
alkyloxyalkyl, alkylthioalkyl, Ar-alkyl or di(Ar)alkyl.
Preferably, the invention relates to compounds of Formula (la) or (Ib) wherein:
R1 is hydrogen, halo, cyano, Ar, Het, alfcyl, and alkyloxy;
p is an integer equal to 1,2, 3 or 4;
R2 is hydrogen, hydroxy, alkyloxy, alkyloxyalkyloxy, alkylthio or a radical
of formula " wherein Y is O ;
R3 is alkyl, Ar, Ar-alkyl or Het ;
R4 is hydrogen, alkyl or benzyl;
R5 is hydrogen, halo or alkyl ; or
two vicinal R5 radicals may be taken together to form together with the phenyl ring to
which they are attached a naphthyl;
r is an integer equal to 1 ; and
R6 is hydrogen ;
R7 is hydrogen or alkyl ;
R8 is oxo ; or
R7 and R8 together form the radical -CH=CH-N=;
alkyl is a straight or branched saturated hydrocarbon radical having from 1 to 6
carbon atoms ; or is a cyclic saturated hydrocarbon radical having from 3 to 6
carbon atoms ; or is a a cyclic saturated hydrocarbon radical having from 3 to 6
carbon atoms attached to a straight or branched saturated hydrocarbon radical
having from 1 to 6 carbon atoms ; wherein each carbon atom can be optionally
substituted with halo or hydroxy ;
Ar is a homocycle selected from the group of phenyl, naphthyl, acenaphthyl,
tetrahydronaphthyl, each optionally substituted with 1, 2 or 3 substituents, each
substituent independently selected from the group of halo, haloalkyl, cyano,
alkyloxy and morpholinyl ;
Het is a monocyclic heterocycle selected from the group of N-phenoxypiperidinyl,
furanyl, thienylj pyridinyl, pyrimidinyl ; or a bicyclic heterocycle selected from
the group of benzothienyl, 2,3-dihydrobenzo[l,4]dioxinyl or benzo[l,3]-
dioxolyl; each monocyclic and bicyclic heterocycle may optionally be
substituted on a carbon atom with 1, 2 or 3 alkyl substituents ; and
halo is a substituent selected from the group of fluoro, chloro and bromo.
For compounds according to either Formula (la) or (Ib), preferably, R1 is hydrogen,
halo, Ar, Het, alkyl or alkyloxy. More preferably, R1 is halo. Most preferably, R1 is
bromo.
Preferably, p is equal to 1.
Preferably, R2 is hydrogen, alkyloxy or alkylthio. More preferably, R2 is alkyloxy.
Most preferably, R2 is methyloxy.
Preferably, R3 is naphthyl, phenyl or Het, each optionally substituted with 1 or 2
substituents, that substituent preferably being a halo or haloalkyl, most preferably being
a halo. More preferably, R3 is naphthyl or phenyl. Most preferably, R3 is naphthyl.
Preferably, R4 is hydrogen or alkyl, more preferably alkyl, such as methyl or ethyl.
Most preferably R4 is methyl
Preferably, R5 is hydrogen, alkyl or halo. Most preferably, R5 is hydrogen.
Preferably r is 1 or 2.
Preferably, R6 is hydrogen or methyl. Most preferably, R6 is hydrogen.
Preferably, Z is CH2.
Preferably, Z is C(=O).
For compounds according to Formula (Ib) only, preferably, R7 is alkyl, preferably
methyl, and R8 is oxygen.
An interesting group of compounds are the compounds of Formula (la), the
pharmaceutically acceptable acid or base addition salts thereof, the quaternary amines
thereof, the stereochemically isomeric forms thereof, the tautomeric forms thereof and
the N-oxide forms thereof.
An interesting group of compounds are those compounds according to Formula (la), the
pharmaceutically acceptable acid or base addition salts thereof, the quaternary amines
thereof, the stereochemically isomeric forms thereof, the tautomeric forms thereof and
the TV-oxide forms thereof, in which R1 is hydrogen, halo, Ar, Het, alkyl or alkyloxy;
p = 1; R2 is hydrogen, alkyloxy or alkylthio; R3 is naphthyl, phenyl or Het, each
optionally substituted with 1 or 2 substituents selected from the group of halo and
haloalkyl; R4 is hydrogen or alkyl; R5 is hydrogen, alkyl or halo; r is equal to 1 and R6
is hydrogen.
An interesting group of compounds are those compounds according to Formula (la)
wherein R1 is hydrogen; halo, e.g. bromo; alkyl, e.g. methyl; or Het, e.g. furanyl; R2 is
alkyloxy, e.g. methyloxy; R3 is naphthyl, phenyl or Het, each optionally substituted
with halo, e.g. phenyl optionally substituted with halo, naphthyl or furanyl; R4 is alkyl,
e.g. methyl or ethyl; R5 is hydrogen or halo, e.g. chloro; R6 is hydrogen; Z is CH2 or
C(=0).
The pharmaceutically acceptable acid addition salts are defined to comprise the
therapeutically active non-toxic acid addition salt forms which the compounds
according to either Formula (la) or (Ib) are able to form. Said acid addition salts can be
obtained by treating the base form of the compounds according to either Formula (la)
or (Ib) with appropriate acids, for example inorganic acids, for example hydrohalic
acid, in particular hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid and
phosphoric acid; organic acids, for example acetic acid, hydroxyacetic acid, propanoic
acid, lactic acid, pyruvic acid, oxalic acid, malonic acid, succinic acid, maleic acid,
ftonaric acid, malic acid, tartaric acid, citric acid, methanesulfonic acid, ethanesulfonic
acid, benzenesulfonic acid, p-toluenesulfonic acid, cyclamic acid, salicyclic acid,
p-aminosalicylic acid and pamoic acid.
The compounds according to either Formula (la) or (Ib) containing acidic protons may
also be converted into their therapeutically active non-toxic base addition salt forms by
treatment with appropriate organic and inorganic bases. Appropriate base salts forms
comprise, for example, the ammonium salts, the alkaline and earth alkaline metal salts,
in particular lithium, sodium, potassium, magnesium and calcium salts, salts with
organic bases, e.g. the benzathine, JV^methyl-D-glucamine, hybramine salts, and salts
with amino acids, for example arginine and lysine.
Conversely, said acid or base addition salt forms can be converted into the free forms
by treatment with an appropriate base or acid.
The term addition salt as used in the framework of this application also comprises the
solvates which the compounds according to either Formula (la) or (Tb) as well as the
salts thereof, are able to form. Such solvates are, for example, hydrates and
alcoholates.
The term "quaternary amine" as used hereinbefore defines the quaternary ammonium
salts which the compounds of formula (la) or (Tb) are able to form by reaction between
a basic nitrogen of a compound of formula (la) or (Tb) and an appropriate quaternizing
agent, such as, for example, an optionally substituted alkylhalide, arylhalide or
arylalkylhalide, e.g. memyliodide or benzyliodide. Other reactants with good leaving
groups may also be used, such as alkyl trifluoromethanesulfonates, alkyl
methanesulfonates, and alkyl p-toluenesulfonates. A quaternary amine has a positively
charged nitrogen. Pharmaceutically acceptable counterions include chloro, bromo,
iodo, trifluoroacetate and acetate. The counterion of choice can be introduced using ion
exchange resins.
The term "stereochemically isomeric forms" as used herein defines all possible
isomeric forms which the compounds of either Formula (la) or (Ib) may possess.
Unless otherwise mentioned or indicated, the chemical designation of compounds
denotes the mixture of all possible stereochemically isomeric forms, said mixtures
containing all diastereomers and enantiomers of the basic molecular structure. More in
particular, stereogenic centers may have the R- or S-configuration; substituents on
bivalent cyclic (partially) saturated radicals may have either the cis- or transconfiguration.
Stereochemically isomeric forms of the compounds of either Formula
(la) or (Ib) are obviously intended to be embraced within the scope of this invention.
Following CAS-nomenclature conventions, when two stereogenic centers of known
absolute configuration are present in a molecule, an R or S descriptor is assigned (based
on Cahtt-Ingold-Prelog sequence rule) to the lowest-numbered chiral center, the
reference center. The configuration of the second stereogenic center is indicated using
relative descriptors [R,R] or [R,S], where R is always specified as the reference
center and [R,R] indicates centers with the same chirality and indicates
centers of unlike chirality. For example, if the lowest-numbered chiral center in the
molecule has an S configuration and the second center is R, the stereo descriptor would
be specified as S-[R,S]. If "a" and "/3" are used: the position of the highest priority
substituent on the asymmetric carbon atom in the ring system having the lowest ring
number, is arbitrarily always in the "a" position of the mean plane determined by the
ring system. The position of the highest priority substituent on the other asymmetriccarbon atom in the ring system relative to the position of the highest priority substituent
on the reference atom is denominated "a", if it is on the same side of the mean plane
determined by the ring system, or "p", if it is on the other side of the mean plane
determined by the ring system.
Compounds of either Formula (la) or (Ib) and some of the intermediate compounds
invariably have at least two stereogenic centers in their structure which may lead to at
least 4 stereochemically different structures.
The compounds of either Formula (la) or (Ib) as prepared in the processes described
below may be synthesized in the form of racemic mixtures of enantiomers which can
be separated from one another following art-known resolution procedures. The
racemic compounds of either Formula (la) or (Ib) may be converted into the
corresponding diastereomeric salt forms by reaction with a suitable chiral acid. Said
diastereomeric salt forms are subsequently separated, for example, by selective or
fractional crystallization and the enantiomers are liberated therefrom by alkali. An
alternative manner of separating the enantiomeric forms of the compounds of either
Formula (la) or (Ib) involves liquid chromatography using a chiral stationary phase.
Said pure stereochemically isomeric forms may also be derived from the corresponding
pure stereochemically isomeric forms of the appropriate starting materials, provided
mat the reaction occurs stereospecifically. Preferably if a specific stereoisomer is
desired, said compound will be synthesized by stereospecific methods of preparation.
These methods will advantageously employ enantiomerically pure starting materials.
The tautomeric forms of the compounds of either Formula (la) or (Ib) are meant to
comprise those compounds of either Formula (la) or (Ib) wherein e.g. an enol group is
converted into a keto group (keto-enol tautomerism).
The JV-oxide forms of the compounds according to either Formula (la) or (Ib) are meant
to comprise those compounds of either Formula (la) or (Ib) wherein one or several
nitrogen atoms are oxidized to the so-called JV-oxide, particularly those JV-oxides
wherein the nitrogen of the amine radical is oxidized.
The invention also comprises derivative compounds (usually called "pro-drugs") of the
pharmacologically-active compounds according to the invention, which are degraded in
vivo to yield the compounds according to the invention. Pro-drugs are usually (but not
always) of lower potency at the target receptor than the compounds to which they are
degraded. Pro-drugs are particularly useful when the desired compound has chemical
or physical properties that make its administration difficult or inefficient. For example,
the desired compound may be only poorly soluble, it may be poorly transported across
the mucosal epithelium, or it may have an undesirably short plasma half-life. Further
discussion on pro-drugs may be found in Stella, V. J. et al, "Prodrugs", Drug Delivery
Systems, 1985, pp. 112-176, and Drugs, 1985,29, pp. 455-473.
Pro-drugs forms of the pharmacologically-active compounds according to the invention
will generally be compounds according to either Formula (la) or (Ib), the
pharmaceutically acceptable acid or base addition salts thereof, the stereochemically
isomeric forms thereof, the tautomeric forms thereof and tibe AT-oxide forms thereof,
having an acid group which is esterified or amidated. Included in such esterified acid
groups are groups of the formula -COORX, where Rx is a alkyl, phenyl, benzyl or
one of the following groups :
Amidated groups include groups of the formula - CONRyR.z, wherein Ry is H,
alkyl, phenyl or benzyl and Rz is -OH, H, alkyl, phenyl or benzyl.
Compounds according to the invention having an amino group may be derivatised with
a ketone, or an aldehyde such as formaldehyde to form a Mannich base. This base will
hydrolyze with first order kinetics in aqueous solution.
The compounds according to the invention have surprisingly been shown to be suitable
for the treatment of mycobacterial diseases, particularly those diseases caused by
pathogenic mycobacteria, including drug resistant and multi drug resistant
mycobacteria, such as Mycobacterium tuberculosis, M. bovis, M. avium, M. smegmatis
andM. marimim. The present invention thus also relates to compounds of either
Formula (la) or (Ib) as defined hereinabove, the pharmaceutically acceptable acid or
base addition salts thereof, the stereochemically isomeric forms thereof, the tautomeric
forms thereof and the AT-oxide forms thereof, for use as a medicine.
The invention also relates to a composition comprising a pharmaceutically acceptable
carrier and, as active ingredient, a therapeutically effective amount of a compound
according to the invention. The compounds according to the invention may be
formulated into various pharmaceutical forms for administration purposes. As
appropriate compositions there may be cited all compositions usually employed for
systemically administering drugs. To prepare the pharmaceutical compositions of this
invention, an effective amount of the particular compound, optionally in addition salt
form, as the active ingredient is combined in intimate admixture with a
pharmaceutically acceptable carrier, which carrier may take a wide variety of forms
depending on the form of preparation desired for administration. These pharmaceutical
compositions are desirable in unitary dosage form suitable, in particular, for
administration orally or by parenteral injection. For example, in preparing the
compositions in oral dosage form, any of the usual pharmaceutical media maybe
employed such as, for example, water, glycols, oils, alcohols and the like in the case of
oral liquid preparations such as suspensions, syrups, elixirs, emulsions and solutions; or
solid carriers such as starches, sugars, kaolin, diluents, lubricants, binders,
disintegrating agents and the like in the case of powders, pills, capsules and tablets.
Because of their ease in administration, tablets and capsules represent the most
advantageous oral dosage unit forms in which case solid pharmaceutical carriers are
obviously employed. For parenteral compositions, the carrier will usually comprise
sterile water, at least in large part, though other ingredients, for example, to aid
solubility, may be included. Injectable solutions, for example, may be prepared in
which the carrier comprises saline solution, glucose solution or a mixture of saline and
glucose solution. Injectable suspensions may also be prepared in which case
appropriate liquid carriers, suspending agents and the like may be employed. Also
included are solid form preparations which are intended to be converted, shortly before
use, to liquid form preparations.
Depending on the mode of administration, the pharmaceutical composition will
preferably comprise from 0.05 to 99 % by weight, more preferably from 0.1 to 70 % by
weight of the active ingredient of formula (la) or (Ib), and, from 1 to 99.95 % by
weight, more preferably from 30 to 99.9 weight % of a pharmaceutically acceptable
carrier, all percentages being based on the total composition.
The pharmaceutical composition may additionally contain various other ingredients
known in the art, for example, a lubricant, stabilising agent, buffering agent,
emulsifying agent, viscosity-regulating agent, surfactant, preservative, flavouring or
colorant.
It is especially advantageous to formulate the aforementioned pharmaceutical
compositions in unit dosage form for ease of administration and unifbrmity of dosage.
Unit dosage form as used herein refers to physically discrete units suitable as unitary
dosages, each unit containing a predetermined quantity of active ingredient calculated
to produce the desired therapeutic effect in association with the required
pharmaceutical carrier. Examples of such unit dosage forms are tablets (including
scored or coated tablets), capsules, pills, powder packets, wafers, suppositories,
injectable solutions or suspensions and the like, and segregated multiples thereof.
The daily dosage of the compound according to the invention will, of course, vary with
the compound employed, the mode of administration, the treatment desired and the
mycobacterial disease indicated. However, in general, satisfactory results will be
obtained when the compound according to the invention is administered at a daily
dosage not exceeding Igram, e.g. in the range from 10 to 50 mg/kg body weight.
Further, the present invention also relates to the use of a compound of either Formula
(la) or (Ib), the pharmaceutically acceptable acid or base addition salts thereof, the
stereochemically isomeric forms thereof, the tautomeric forms thereof and the .W-oxide
forms thereof, as well as any of the aforementioned pharmaceutical compositions
thereof for the manufacture of a medicament for the prevention or the treatment of
mycobacterial diseases.
Accordingly, hi another aspect, the invention provides a method of treating a patient
suffering from, or at risk of, a mycobacterial disease, which comprises administering
to the patient a therapeutically effective amount of a compound or pharmaceutical
composition according to the invention.
The compounds of the present invention may also be combined with one or more other
antimycobacterial agents.
Therefore, the present invention also relates to a combination of (a) a compound of
formula (la) or (Ib) and (b) one or more other antimycobacterial agents.
The present invention also relates to a combination of (a) a compound of formula (la)
or (Ib) and (b) one or more other antimycobacterial agents for use as a medicine.
A pharmaceutical composition comprising a pharmaceutically acceptable carrier and,
as active ingredient, a therapeutically effective amount of (a) a compound of formula
(la) or (Ib) and (b) one or more other antimycobacterial agents is also comprised by the
present invention.
The other Mycobacterial agents which may be combined with the compounds of
formula (la) or (Ib) are for example rifampicin (=rifampin); isoniazid; pyrazinamide;
amikacin; ethkmamide; moxifloxacin; ethambutol;. streptomycin; para-aminosalicylic
acid; cycloserine; capreomycin; kanamycin; thioacetazone; PA-824;
quinolones/fluoroquinolones such as for example ofloxacin, ciprofloxacin,
sparfloxacin; macrolides such as for example clarithromycin, clofazimine, amoxycillin
with clavulanic acid; rifamycins; rifabutin; rifapentine.
Preferably, the present compounds of formula (la) or (Ib) are combined with rifapentin
and moxifloxacin.
GENERAL PREPARATION
The compounds according to the invention can generally be prepared by a succession
of steps, each of which is known to the skilled person.
Compounds of formula (la) and (Ib) wherein Z is CHa, said compounds being
represented by formula (Ia-1) and (Tb-1), can be prepared by reacting an intermediate of
formula (Il-a) and (n-b) with paraformaldehyde in a suitable solvent, such as for
example toluene.
Compounds of formula (la) and (Tb) wherein Z is C(=O), said compounds being
represented by formula (Ia-2) and (Tb-2), can be prepared by reacting an intermediate of
fonnula (Hl-a) and (lH-b) wherein represents a suitable leaving group, such as for
example imidazole, alkoxy groups, e.g. methoxy, with a suitable base, such as for
example sodium hydride, potassium tertiobutylate, in a suitable solvent, such as for
example tetrahydrofuran, diethylether, dioxane.
In the above reactions, the obtained compound of formula (la) or (Ib) can be isolated,
and, if necessary, purified according to methodologies generally known in the art such
as, for example, extraction, crystallization, distillation, trituration and chromatography.
In case the compound of formula (la) or (Ib) crystallizes out, it can be isolated by
filtration. Otherwise, crystallization can be caused by the addition of an appropriate
solvent, such as for example water; acetonitrile; an alcohol, such as for example
methanol, ethanol; and combinations of said solvents. Alternatively, the reaction
mixture can also be evaporated to dryness, followed by purification of the residue by
chromatography (e.g. reverse phase HPLC, flash chromatography and the like). The
reaction mixture can also be purified by chromatography without previously
evaporating the solvent. The compound of formula (la) or (Ib) can also be isolated by
evaporation of the solvent followed by recrystallisation in an appropriate solvent, such
as for example water; acetonitrile; an alcohol, such as for example methanol; and
combinations of said solvents.
The person skilled in the art will recognise which method should be used, which
solvent is the most appropriate to use or it belongs to routine experimentation to find
the most suitable isolation method
The compounds of formula (la) or (Ib) may further be prepared by converting
compounds of formula (la) or (Ib) into each other according to art-known group
transformation reactions.
The compounds of formula (la) or (Ib) may be converted to the corresponding N-oxide
forms following art-known procedures for converting a trivalent nitrogen into its
JV-oxide form. Said JV-oxidation reaction may generally be carried out by reacting the
starting material of formula (la) or (Ib) with an appropriate organic or inorganic
peroxide. Appropriate inorganic peroxides comprise, for example, hydrogen peroxide,
alkali metal or earth alkaline metal peroxides, e.g. sodium peroxide, potassium
peroxide; appropriate organic peroxides may comprise peroxy acids such as, for
example, benzenecarhoperoxoic acid or halo substituted benzenecarboperoxoic acid,
e.g. 3-chlorobenzenecarboperoxoic acid, peroxoalkanoic acids, e.g. peroxoacetic acid,
alkylhydroperoxides, e.g. tbutyl hydro-peroxide. Suitable solvents are, for example,
water, lower alcohols, e.g. ethanol and the like, hydrocarbons, e.g. toluene, ketones,
e.g. 2-butanone, halogenated hydrocarbons, e.g. dichloromethane, and mixtures of such
solvents.
Compounds of formula (Ta) or (Ib) wherein R4 is alkyl, may be converted into a
suitable quaternary amine by reaction with a suitable quatemizing agent, such as, for
example, an optionally substituted alkylhalide, e.g. ICHg, in the presence of a suitable
solvent, such as for example acetone.
Some of the compounds of formula (I) and some of the intermediates in the present invention
may consist of a mixture of stereochemically isomeric forms. Pure
stereochemically isomeric forms of said compounds and said intermediates can be
obtained by the application of art-known procedures. For example, diastereoisomers
can be separated by physical methods such as selective crystallization or
chromatographic techniques, e.g. counter current distribution, liquid chromatography
and the like methods. Enantiomers can be obtained from racemic mixtures by first
converting said racemic mixtures with suitable resolving agents such as, for example,
chiral acids, to mixtures of diastereomeric salts or compounds; then physically
separating said mixtures of diastereomeric salts or compounds by, for example,
selective crystallization or chromatographic techniques, e.g. liquid chromatography and
the like methods; and finally converting said separated diastereomeric salts or
compounds into the corresponding enantiomers. Pure stereochemically isomeric forms
may also be obtained from the pure stereochemically isomeric forms of the appropriate
krtermediates and starting materials, provided that the intervening reactions occur
stereospecifically.
An alternative manner of separating the enantiomeric forms of the compounds of
formula (I) and intermediates involves liquid chromatography, in particular liquid
chromatography using a chiral stationary phase.
It is to be understood that in the above or the following preparations, the reaction
products may be isolated from the reaction medium and, if necessary, further purified
according to methodologies generally known in the art such as, for example, extraction,
crystallization, distillation, trituration and chromatography.
Some of the intermediates and starting materials are known compounds and may be
commercially available or may be prepared according to art-known procedures.
Intermediates of formula (H-a) and (II-b) can be prepared by reacting an intermediate
of formula (IV-a) and (W-b) with a suitable deprotecting agent, such as for example
1 -chloroethyl chloroformate, in a suitable solvent, such as for example
1,2-dichloroethane and a suitable alcohol, such as for example methanol and the like.
Intermediates of formula (Tl-a) or (II-b) can also be prepared by reacting an
intermediate of formula (IV-a) or (TV-b) with ammonium formate in the presence of
palladium on charcoal and in the presence of a suitable solvent, such as for example an
alcohol, e.g. methanol. Intermediates of formula (TV-a) or (TV-b) wherein R1 is halo,
may loose said halo substituent during their transformation to intermediates of formula
(H-a) or (n-b).
Intermediates of formula (IV-a) and (IV-b) can be prepared by reacting an intermediate
of formula (V-a) and (V-b) with an intermediate of formual (VT) in the presence of a
suitable reducing agent, such as for example n-BuLi, in the presence of a suitable base,
such as for example JNTV-diisopropylamine, and in the presence of a suitable solvent,
such as for example tetrahydrofuran.
Intermediates of formula (IV-a) or (IV-b) wherein R1 represents Het and p is 1, said
intermediates being represented by formula (IV-a-1) or (IV-b-1), can be prepared by
reacting an intermediate of formula (TV-a) or (TV-b) wherein R1 represents halo, said
intermediates being represented by formula (IV-a-2) or (IV-b-2), with an intermediate
of formula (VII) in the presence of a suitable catalyst, such as for example Pd(PPh3)4, a
suitable base, such as for example KjCOs, and a suitable solvent, such as for example
dimethylether and a suitable alcohol, such as for example methanol and the like.
Intermediates of formula (m-a) or (m-b) can be prepared by reacting an intermediate
of formula (Il-a) or (TI-b) with an intermediate of formula (VIE) wherein Wi-(C=O)
represents the group that has to be introduced and R represents the remaining of the
intermediate, such as for example l,l'-carbonylbis-l-imidazole,
methylchloroformiate or ethylchloroformiate, in the presence of a suitable solvent, such
as for example tetrahydrofioran.
The intermediate compounds of formula (V-a) or (V-b) are compounds that are either
commercially available or may be prepared according to conventional reaction
procedures generally known in the art. For example, intermediates of formula (V-a-1)
may be prepared according to the following reaction scheme (1):
wherein all variables are defined as in Formula (la) and (Ib). Reaction scheme (1)
comprises step (a) in which an appropriately substituted aniline is reacted with an
appropriate acylchloride such as 3-phenylpropionyl chloride, 3-fhiorobenzenepropionyl
chloride orp-chlorobenzenepropionyl chloride, in the presence of a suitable base, such
as triethylamine and a suitable reaction-inert solvent, such as methylene chloride or
ethylene dichloride. The reaction may conveniently be carried out at a temperature
ranging between room temperature and reflux temperature. In a next step (b) the
adduct obtained in step (a) is reacted with phosphoryl chloride (POCla) in the presence
of a suitable solvent, such as for example-AAT-diniethylformamide (Vilsmeier-Haack
formylation followed by cyclization). The reaction may conveniently be carried out at
a temperature ranging between room temperature and reflux temperature. In a next
step (c) a specific R2-group, wherein R2 is an alkyloxy or alkylthio radical is introduced
by reacting the intermediate compound obtained in step (b) with a compound -X-Alk,
wherein X=S or O and Alk is an alkylgroup as defined in Formula (la) and (Ib), such as
for example sodium methanolate, in the presence of a suitable solvent, such as for
example an alcohol, e.g. methanol.
Intermediates according to Formula (V-a-2) may be prepared according to the
following reaction scheme (2), wherein in a first step (a) a substituted indole-2,3-dione
is reacted with a substituted 3-phenylpropionaldehyde in the presence of a suitable base
such as sodium hydroxide (Pfitzinger reaction), after which the carboxylic acid
compound in a next step (b) is decarboxylated at high temperature in the presence of a
suitable reaction-inert solvent such as diphenylether.
It is evident that in the foregoing and in the following reactions, the reaction products
may be isolated from the reaction medium and, if necessary, further purified according
to methodologies generally known in the art, such as extraction, crystallization and
chromatography. It is further evident that reaction products that exist in more than one
enantiomeric form, may be isolated from their mixture by known techniques, in
particular preparative chromatography, such as preparative HPLC. Typically,
compounds of Formula (la) and (Ib) maybe separated into their isomeric forms.
The intermediates of Formula (VI) are compounds that are either commercially
available or may be prepared according to conventional reaction procedures generally
known in the art For example, intermediate compounds of Formula (VI) may be
prepared according to the following reaction scheme (3):
Reaction scheme (3) comprises step (a) in which R3, for example an appropriately
substituted phenyl, naphthyl, or Het, is reacted by Friedel-Craft reaction with an
appropriate acylchloride such as 3-chloropropionyl chloride or 4-chlorobutyryl
chloride, in the presence of a suitable Lewis acid, such as AlCla, FeCla, SnCLt, TiCL or
ZnCla and optionally a suitable reaction-inert solvent, such as rnethylene chloride or
ethylene dichloride. The reaction may conveniently be carried out at a temperature
ranging between room temperature and reflux temperature. In a next step (b) an araino
group (-NR^CEfc-CeHs) is introduced by reacting the intermediate compound obtained
in step (a) with a primary or secondary amine, in the presence of a suitable solvent,
such as for example acetonitrile, and optionally in the presence of a suitable base, such
as for example KaCOa.
The following examples illustrate the present invention without being limited thereto.
EXPERIMENTAL PART
Of some compounds the absolute stereochemical configuration of the stereogenic
carbon atom(s) therein was not experimentally determined. In those cases the
stereochemically isomeric form which was first isolated is designated as "A" and the
second as "B", without further reference to the actual stereochemical configuration.
However, said "A" and "B" isomeric forms can be unambiguously characterized by a
person skilled in the art, using art-known methods such as, for example, X-ray
diffraction. The isolation method is described in detail below.
For some of the intermediates and some of the final compounds, stereochemical
configurations are indicated in the structures. These configurations are relative
configurations indicating that the groups concerned are located in the same or opposite
= opposite plane
plane of the molecule
R means that the chiral center is absolute R or absolute S.
S means that the chiral center is absolute R or absolute S.
Experimental part
Hereinafter, the term 'M.P." means melting point, 'THF' means tetrahydrofuran,
'EtOAc' means ethyl acetate, 'MeOH' means methanol, 'DME' means dimethyl ether,
'DIPE' means diisopropyl ether, 'DMF' means N,N-dimethylformamide, "EtsN" means
triethylamine, 'Pd(PPh3)4 means tetralds(triphenylphosphine)palladium, 'CDF means
1,1 '-carbonylbis-lH-imidazole.
A. Preparation of the intermediates
Example Al
Preparation of intermediate 1
Br.
Intermediate 1
Benzenepropanoylchloride (0.488 mol) was added dropwise at room temperature to a
solution of 4-bromobenzenamine (0.407 mol) in Et3N (70ml) and CHaCl2 (700 ml) and
the mixture was stirred at room temperature overnight The mixture was poured out
into water and concentrated NELtOH, and extracted with CHaCla. The organic layer
was dried (MgSO/t), filtered, and the solvent was evaporated. The residue was
crystallized from diethyl ether. The residue (119.67 g) was taken up in CHaCk and
washed with HC1 IN . The organic layer was dried (MgSC4), filtered, and the solvent
was evaporated. Yield: 107.67 g of intermediate 1.
Example A2
Preparation of intermediate 2
Intermediate 2
The reaction was carried out twice. POCla (1.225 mol) was added dropwise at 10 °C
to DMF (0.525 mol). Then intermediate 1 (0.175 mol) was added at room temperature
. The mixture was stirred overnight at 80 °C, poured out on ice and extracted with
CHaCfe. The organic layer was dried (MgSO4), filtered, and the solvent was
evaporated, yielding 77.62 g (67%) of intermediate 2. The product was used without
further purification.
Example A3
Preparation of intermediate 3
Intermediate 3
A mixture of intermediate 2 (0.233 mol) in CH3ONa (30%) in MeOH (222.32 ml) and
MeOH (776 ml) was stirred and refluxed overnight, then poured out on ice and
extracted with ClfcCla The organic layer was separated, dried (MgSO4), filtered and
the solvent was evaporated. The residue was purified by column chromatography over
silica gel (eluent: CHaCVcyclohexane 20/80 and then 100/0; 20-45um). The pure
fractions were collected and the solvent was evaporated. Yield: 25 g (33%) of
intermediate 3 (M.P.: 84 °C).
Example A4
a) Preparation of intermediates 4 and 5
Intermediate 4 Intermediate 5
A mixture of aluminium chloride (34.3 g, 0.257 mol) and 3-chloropropanoyl chloride
(29.7 g, 0.234 mol) in 1,2-dichloroethane (150 ml) was stirred at 0°C. A solution of
naphthalene (30 g, 0.234 mol) in 1,2-dichloroethane (50 ml) was added. The mixture
was stirred at 5 °C for 2 hours and poured out into ice water. The organic layer was
separated, dried (MgSO4), filtered, and the solvent was evaporated. The residue (56 g)
was purified by column chromatography over silica gel (eluent: cyclohexane/ CHbCla :
60/40; 20-45um). Two fractions were collected and the solvent was evaporated to
afford intermediate 4 (31 g, 61%) as an oil. The second fraction (14 g) was taken up in
DEPE to afford intermediate 5 (8.2 g, 16%; M.P.: 68 °C) as a pale yellow solid.
Following intermediate was prepared according to the previous procedure:
The residue (20.0 g) was used for the
Intermediate 41
next step without further purification.
Intermediate 41
b) Preparation of intermediate 6
Intermediate 6
Aluminum chloride (0.3 mol) was added carefully to 1,3-difluorobenzene (0.26 mol)
and they were heated with vigorously stirring till 50 °C. 3-chloropropanoyl chloride
(0.26 mol) was added dropwise over a 15 minute period at 40 °C (cooled on ice) and
the mixture was stirred at 50 °C. The mixture was poured into water (250 ml), ice (250
g) andiHCl (25 ml) and it was stirred for 20 minutes The formed precipitate was
filtered off and extracted with CH2C12 and water. Yield: 40 g of intermediate 6 (75 %).
Example AS
a) Preparation of intermediate 7
Intermediate 7
A mixture of intermediate 4 (3 g; 0.0137 mol), N-benzyhnethyl amine (2 ml; 0.0150
mol) in acetonitrile (100 ml) was stirred at 80 °C for 2 hours. At room temperature
water was added. The mixture was extracted with CHaCla . The organic layer was
separated and dried (MgSO4), filtered, and the solvent was evaporated. The residue (6
g) was purified by column chromatography over silica gel (eluent:
97/3; 20-45um) to afford an oil. Yield: 4.2 g of intermediate 7.
Following intermediate was prepared according to the previous procedure:
Intermediate 42
The residue (22.5 g) was purified by
column chromatography over silica
gel (eluent: CH2Cl2/MeOH: 98/2; 20-
45um) to afford an oil. Yield: 5.1 g of
intermediate 42 (17%).
b) Preparation of intermediate 8
Intermediate 8
A mixture of intermediate 6 (0.015 mol), N-ethylbenzenemethanamine (0.016 mol) and
K2CO3 (0.016 mol) in acetonitrile (30 ml) was stirred at 70 °C for 2 hours, poured out
into HaO and extracted with CEfcCk. The organic layer was separated, dried
filtered, and the solvent was evaporated. Yield: 4 g of intermediate 8 (88%).
Example A6
a) Preparation of intermediate 9
Intermediate 9
n-Butyl lithium (0.0075 mol) was added at —20 °C to a solution of diisopropylamine
(0.0075 mol) in THF (50ml). The mixture was cooled to -70 °C. Intermediate 3
(0.0062 mol) was added. The mixture was stirred at -70 °C for 1 hour and 30 minutes.
Intermediate 7 (0.0075 mol) was added. The mixture was stirred for 1 hour and 30
minutes. HjO was added. The mixture was extracted with CHaCfc. The organic layer
was separated, dried (MgSCU), filtered, and the solvent was evaporated. The residue (3
g) was purified by column chromatography over silica gel (eluent: cyclohexane/EtOAc
90/10; 15-40um). The pure fractions were collected and the solvent was evaporated.
Yield: 1.5 g of a mixture of two diastereoisomers (38%), i.e. intermediate 9.
Following intermediate was prepared according to the previous procedure:
Intermediate 43
The residue (7.5 g) was purified by
column chromatography over silica
gel (eluent: cyclohexane/EtOAc 92/8;
15-40|jjn). The pure fractions were
collected and the solvent was
evaporated. Yield: 3.25 g of
intermediate 43 a mixture of two
diastereoisomers (55%, mixture of
diastereoisomers: 65/35).
b) Preparation of intermediate 10
Intermediate 10
n-Butyl lithium (0.0075 mol) was added at -20 °C to a solution of diisopropylamine
(0.0075 mol) in THF (50ml). The mixture was cooled to -70 °C. Intermediate 3
(0.0061 mol) was added. The mixture was stirred at -70 °C for 1 hour and 30 minutes.
4-[methyl(phenyhiiethyl)amino]-l-phenyl- 1-butanone (0.0073 mol) was added. The
mixture was stirred for 1 hour and 30 minutes. EfeO was added. The mixture was
extracted with CHaQz. The organic layer was separated, dried (MgSO-O, filtered, and
the solvent was evaporated. The residue (4.9 g) was purified by column
chromatography over silica gel (eluent: 100% CBbGb; 15-40um). The pure fractions
were collected and the solvent was evaporated, yielding: 1.43 g of a intermediate 10
(40%, mixture of diastereoisomers: 60/40).
Following intermediates were according to the previous procedure:
Intermediate 19 The residue (6.4 g) was purified by
column chromatography over silica
gel (eluent: CEfeCWcyclohexane
85/15; 15-40um). The pure fractions
were collected and the solvent was
evaporated, yielding 0.81 g of the
intermediate as a mixture of
diastereoisomers (44/56) (17%).
Intermediate 20 The residue was purified by column
chromatography over silica gel
(eluent: cyclohexane/EtOAc 95/5; 15-
40um). The pure fractions were
collected and the solvent was
evaporated. Yield: 0.55 g of the
intermediate as a mixture of
diastereoisomers (12%).
Intermediate 20
Intermediate 21 The residue was purified by column
chromatography over silica gel
(eluent: cyclohexane/ EtOAc 95/5; 15-
40um). The pure fractions were
collected and the solvent was
evaporated, to give 0.34 g of the
intermediate as a mixture of
diastereoisomers (7%).
Intermediate 22 The residue was purified by column
chromatography over silica gel
(eluent: cyclohexane/ EtOAc 95/5; 15-
40um). The pure fractions were
collected and the solvent was
evaporated, yielding 0.80 g of the
intermediate as a mixture of
diastereoisomers (13 %).
Intermediate 22
Tntermediate 23 The residue was purified by column
chromatography over silica gel
(eluent: cyclohexane/ EtOAc 80/20;
15-40 jam).). The pure fractions were
collected and the solvent was
evaporated. The residue (1.3 g) was
purified by column chromatography
over silica gel (eluent:
acetonitrile/NBUCOs 0.5% 95/5;
kromasil). The pure ftactions were
collected and the solvent was
evaporated, affording 0.61 g of the
intermediate as a mixture of
diastereoisomers (41/59) (18 %).
Intermediate 23
c) Preparation of intermediates 11 and 12
Intermediate 11 (dia A) and
Intermediate 12 (dia B)
n-Buryl lithium (0.0075 mol) was added at—20 °C to a solution of diisopropylamine
(0.0075 mol) in THF (50 ml). The mixture was cooled to -70 °C. Intermediate 3
(0.00824 mol) was added. The mixture was stirred at -70 °C for 1 hour and 30 minutes.
Intermediate 8 (0.0099 mol) was added. The mixture was stirred for 1 hour and 30
minutes. HaO was added. The mixture was extracted with CEbQb, The organic layer
was separated, dried (MgSO4), filtered, and the solvent was evaporated. The residue
(5.4 g) was purified by column chromatography over silica gel (eluent:
CEbCb/cyclohexane 60/40 ; 15-40um). Two fractions were collected and the solvent
was evaporated. Yielding: 0.95g of intermediate 11 as diastereoisomer A (15%, M.P.:
171 °C) and0.83g of intermediate 12 as diastereoisomer B (13%, MH+: 631).
Example A7
Preparation of intermediate 17
Intermediate 17
Intermediate 9 (1.58 mmol), 2-furanboronic acid (2.69 mmol), Pd(PPh3)4 (0.158
mmol), DME (30 ml), MeOH (10 ml) and K2CO3 (1.6 ml) were heated under
microwaves (300W, 68 °C) for 10 minutes. The mixture was cooled, poured into water
and extracted with EtOAc. The organic layer was separated, dried (MgSCU), filtered,
and the solvent was evaporated. The residue (1.4 g) was purified by column
chromatography over silica gel (eluent: cyclohexane/EtOAc 90/10; 15-40um). The pure
fractions were collected and the solvent was evaporated. Yield: 0.47 g of intermediate
17 as a mixture of diastereoisomers (60/40) (41%).
Example A8
a-1) Preparation of intermediates 13 and 14
Intermediate 13 (diastereoisomer A) Intermediate 14 (diastereoisomer B)
1-Chloroethyl chloroformate (15 ml) was added at room temperature to a mixture of
intermediate 9 (0.0023 mol) in 1,2-dichloroethane (30 ml). The mixture was stirred at
80 °C for 1 hour. The solvent was evaporated. MeOH (15 ml) was added. The mixture
was stirred and refluxed for 30 minutes. The solvent was evaporated. The residue (*)
(1.49 g) was purified by column chromatography over silica gel (eluent:
CHaCla/MeOH/NKUOH 97/3/0.1; 15-40um). Two fiactions were collected and the
solvent was evaporated. The first residue (0.23 g) was crystallized from DIPE. The
precipitate was filtered off and dried, yielding: 0.168 g (13%) of intermediate 13
(diastereoisomer A) (M.P.: 204 °C).The second residue (0.32 g) was crystallized from
DIPE. The precipitate was filtered off and dried. Yield : 0.298 g (23%) of intermediate
14 (diastereoisomer B) (M.P.: 225 °C).
Following intermediates were prepared according to the above procedure. The
purification of the resulting residue is indicated for each intermediate separately.
Intermediate 25
and
Intermediate 26
The residue (1,2 g) was purified by
column chromatography over silica
95/5/0.1; 15-40fjm). Two fractions
were collected and the solvent was
evaporated.
Yield : 0.047 g of intermediate 25
(diastereoisomer A) (6%, MH+ : 491).
The second residue (0.08 g, 10%) was
crystallized from DIPE. The
precipitate was filtered off and dried.
Yield : 0.031 g of intermediate 26
(diastereoisomer B) (4%, M.P.: 197
Intennediate 25 (diastereoisomer A)
Intermediate 26 (diastereoisomer B)
Intermediate 27
and
Intermediate 28
The residue (1.0 g) was purified by
column chromatography over silica
gel (eluent: CCk/MeOH/NEUOH
96/4/0.1; 15-40|jm). Two fractions
were collected and the solvent was
evaporated. Yield: 0.105 g of
intennediate 27 (diastereoisomer A)
(15%, MH+: 539) and 0.11 g of
intennediate 28 (diastereoisomer B)
(16%,M.P.:222°C).
Intennediate 27 (diastereoisomer A)
Intermediate 28 (diastereoisomer B)
Intermediate 29
and
Intermediate 30
The residue (0.5g) was purified by
column chromatography over silica
gel (eluent: OHaCla/MeOH/NKtOH
97/3/0.1; lOum). Two fractions were
collected and the solvent was
evaporated. Yield: 0.11 g of
intermediate 29 (diastereoisomer A)
(24%, MH+: 497) and 0.10 g of
intermediate 30 (diastereoisomer B)
(22%,MH+:497).
Intermediate 29 (diastereoisomer A)
Intermediate 30 (diastereoisomer B)
Intermediate 31
and
Intermediate 32
The residue (0.32 g) was purified by
column chromatography over silica
gel (eluent: CHaCla/MeOH/NHOH
97/3/0.1; lOum). Two fractions were
collected and the solvent was
evaporated. Yield: 0.10 g of
intermediate 31 (diastereoisomer A)
(35%, MH+: 493) and 0.04 g of
intermediate 32 (diastereoisomer B)
(14%, MH+,.: 493).
Intermediate 31 (diastereoisomer A)
Intermediate 32 (diastereoisomer B)
Intermediate 33
and
Intermediate 34
The residue (0.9 g) was purified by
column chromatography over silica
98/2/0.1; lOum). Two fractions were
collected and the solvent was
evaporated. Yield : 0.09 g of
intermediate 33 (diastereoisomer A)
(15%, MH+ : 477) and 0.08 g of
intermediate 34 (diastereoisomer B)
(13%,MH+:477).
Intermediate 33 (diastereoisomer A)
Intermediate 34 (diastereoisomer B)
Intermediate 35
and
Intermediate 36
The residue (0.45 g) was purified by
column chromatography over silica
gel (eluent: C3J2a2/MeOH/NH4OH
96/4/0.1; lOum). Two fractions were
collected and the solvent was
evaporated. Yield: 0.09 g of
intermediate 35 (diastereoisomer A)
(22%, MH+: 529) and 0.12 g of
intermediate 36 (diastereoisomer B)
(30%,MH+:529).
Intermediate 35 (diastereoisomer A)
Intermediate 36 (diastereoisomer B)
Intermediate 37
and
Intermediate 38
The residue (0.63 g) was purified by
column chromatography over silica
gel (eluent: OKaCkMeOH/NBLtOH
98/2/0.1; 15-40um). Two fractions
were collected and the solvent was
evaporated. Yield: 0.08 g of
intermediate 37 (diastereoisomer A)
(15%, MH+ : 575) and 0.06 g of
intermediate 38 (diastereoisomer B)
(12%,MH+:575).
Intermediate 37 (diastereoisomer A)
Intermediate 38 (diastereoisomer B)
Intermediate 39 The residue (0.92 g) was purified by
column chromatography over silica
gel (eluent: CH2Cl2/MeOH 95/5; 15-
40um). The pure fractions were
collected and the solvent was
evaporated. Yielding: 0.4g of
intermediate 39 (diastereoisomer A)
(56%, MH+ : 541).
Intermediate 39 (diastereoisomer A)
Intermediate 40 The residue (0.49 g) was purified by
column chromatography over silica
gel (eluent: CHaCb/MeOH/NBUOH
96/4/0.1; 15-40um). The pure
fractions were collected and the
solvent was evaporated. Yielding:
0.265g of intermediate 40
(diastereoisomer B) (66%, MH+:
541).
Intermediate 40 (diastereoisomer B)
a-2) Preparation of intermediates 15 and 16
Intermediate 15 Intermediate 16
Intermediate 13 (diastereoisomer A) (0.9 g) was purified by chiral chromatography
over silica gel (eluent: 100% ethanol). Two fractions were collected and the solvent
was evaporated. Yield: 0.420 g of intermediate 15 (enantiomer Al) (M.P.: 161 °C,
MEN-: 541) and 0.397g of intermediate 16 (enantiomer A2) (M.P.: 158 °C, MH+:
541).
a-3) Preparation of intermediates 44 and 45
Intermediate 44 Intermediate 45
A mixture of intermediate 43 (prepared according to A6.a) (1.5 g, 2.62 mol),
ammonium formate (0.83 g, 0.013 mol) and palladium on charcoal (10%, 1.5 g) in
methanol (30 ml) was heated under reflux for 1 hour. The mixture was cooled and
filtered on a short pad of celite. Water was added. The organic layer was extracted
with ethyl acetate, separated, dried (MgSO4), filtered, and the solvent was evaporated.
The residue (1.3 g) was purified by column chromatography over silica gel (eluent:
MeOH/AcNELj: 60/40, kromasil Cjg, 5 |Hn) The pure fractions were collected and the
solvent was evaporated yielding two fractions. Yield: 0.14 g of intermediate 44 as
diastereoisomer A (12%) and 0.26 g of intermediate 45 as diastereoisomer B (22%).
Example A9
Preparation of intermediate 18
Intermediate 18
A mixture of the intermediate 39 prepared according to example A8.a-l) (0.0002 mol)
and GDI (0.0003 mol) in THF (7 ml) was stirred and refluxed for 2 hours, poured out
into HaO and extracted with CHaCfc. The organic layer was separated, dried (MgSO4),
filtered, and the solvent was evaporated. Yield: 0.15 g of intermediate 18
(diastereoisomer A) (84%).
B. Preparation of the compounds
Example Bl
Preparation of compound 1
Compound 1
A mixture of intermediate 13 (prepared according to example A8. a-1) (0.00009 mol)
andparaformaldehyde (0.0001 mol) in toluene (5 ml) was stirred at 80 °C. The mixture
was evaporated. The residue was purified by column chromatography over silica gel
(eluent: CH2C32/M:eOH 99/1; 15-40 )j,m). The pure fractions were collected and the
solvent was evaporated. Yield: 0.025 g of compound 1 (diastereoisomer A)
(49%,M.P.:112°C).
Following compounds were prepared according to the above procedure. The
purification of the residue is indicated if different from the above-described
purification.
Compound 6
0.068 g of diastereoisomer A (69 %,
MH+: 551)
Compoimd 6 (diastereoisomer A)
Compound 7
0.11 gof diastereoisomer A (98 %,
MH+: 509)
Compound 7 (diastereoisomer A)
Compounds
0.08 g of diastereoisomer A (80 %,
MH+: 505)
Compound 8 (diastereoisomer A)
Compound 9
0.082 g of diastereoisomer A (100 %,
MB+: 489)
Compound 9 (diastereoisomer A)
Compound 10
0.082 g of diastereoisomer A (89 %,
MH+: 541)
Compound 10 (diastereoisomer A)
Compound 11
The residue was purified by column
chromatography over silica gel
(eluent: CH2Cl2/MeOH99/l; 15-40
|j,m). The pure fiactions were collected
and the solvent was evaporated. Yield:
0.036 g of diastereoisomer B (71%,
M.P.: 108 °C).
Compound 11 (diastereoisomer B)
Compoundl2
0.045 g of diastereoisomer B (88 %,
M.P.: 168 °C)
Compound 12 (diastereoisomer B)
Compound 13
0.077 g of diastereoisomer B (61 %,
MH+: 551).
Compound 13 (diastereoisomer B)
Compound 14
0.040 g of diastereoisomer B (100 %,
MH-f: 505).
Compound 14 (diastereoisomer B)
Compound. 15
0.044 g of diastereoisomer B (72 %,
MH+: 489).
Compound 15 (diastereoisomer B)
Compound 16
012 g of diastereoisomer B (98 %,
MH+: 541).
Compound 16 (diastereoisomer B)
Compound 17
0.034 g of diastereoisomer B (90 %,
MH+: 587).
Compound 17 (diastereoisomer B)
Example
Preparation of compound 2
Compound 2
A mixture of intermediate 37 (diastereoisomer A prepared according to example A8.a-
1) (0.00009 mol) andparaformaldehyde (0.0001 mol) in toluene (5 ml) was stirred at
80 °C. The mixture was evaporated. The residue was purified by column
chromatography over silica gel (eluent: CHaCVMeOH 99/1; 15-40 urn). The pure
fractions were collected and the solvent was evaporated. The residue (0.06 g, 92%) was
crystallized from DEPE. The precipitate was filtered off and dried. Yield: 0.026 g of
compound2 (diastereoisomer A) (40%, M.P.: 201°C).
Example B3
Preparation of compound 3
Compound 3
A mixture of intermediate 30 (diastereoisomer B prepared according to example A8.a-
1) (0.00009 mol) andparaformaldehyde (0.0001 mol) in toluene (5 ml) was stirred at
80 °C. The mixture was evaporated. The residue was purified by column
chromatography over silica gel (eluent: CH2Cl2/MeOH 99/1; 15-40 |im). The pure
fractions were collected and the solvent was evaporated. The residue (0.11 g, 100%)
was crystallized from diethyl ether. The precipitate was filtered off and dried. Yielding
: 0.033 g of compound 3 (diastereoisomer B ) (33%, M.P.: 189 °C).
Example B4
Preparation of compound 4
Compound 4
A mixture of compound 7 (diastereoisomer A prepared according to example B1)
(O.lmmol) and iodomethane (0.1 mmol) in acetone (2 ml) was stirred at room
temperature for 2.5 hours. The precipitate was filtered, washed with acetone and dried.
Yield: 0.031 g compound 4 as a hydroiodide (diastereoisomer A) (48%, M.P.: 211 °C)
Following compound was prepared according to the previous procedure:
Compound 18
0.046 g of diastereoisomer B as a
hydroiodide (71 %, M.P.: 195 °C).
Compound 18 (diastereoisomer B)
Example B5
Preparation of compound 5
Compound 5
Sodium hydride (0.011 g) was added at 0 °C to a mixture of intermediate 18 (prepared
according to example A9) (0.0001 mol) in THF (10 ml) under N2 flow. The mixture
was stirred at 0 °C for 30 minutes, poured out into EkO and extracted with EtOAc. The
organic layer was separated, dried (MgSC4), filtered, and the solvent was evaporated.
The residue was purified by column chromatography over silica gel (eluent:
cyclohexane/EtOAc 80/20; 15-40um). The pure fractions were collected and the
solvent was evaporated. The residue (0.09 g, 67%) was crystallized from DIPE. The
precipitate was filtered off and dried Yield: 0.034 g of compound 5
(diastereoisomer A) (M.P.: 192 °C).
Following compound was prepared according to the previous procedure:
Compound 19
0.047g of diastereoisomer B (M.P.
216 °C).
Compound 19 (diastereoisomer B)
Example B6
Preparation of compound 20
Compound 20
A mixture of intermediate 44 (diastereoisomer A prepared according to example A8a-
3) (0.12 g, 0.298 mmol) andparaformaldehyde (0.358 mmol) in toluene (5 ml) was
stirred at 80°C for 2 hours. Water was added at room temperature and the organic layer
was extracted with ethyl acetate, separated, dried (MgSO/i), filtered, and the solvent
was evaporated. The residue (0.13 g) was purified by column chromatography over
silica gel (eluent: CH2C12/CH3OH/NH4OH : 99/1/0.1, lOum). The pure fractions were
collected and the solvent was evaporated. Yield: 0.075 g of compound 20
(diastereoisomer A ) (61 %, MH+: 415).
Following compound was prepared according to the previous procedure but without the
column chromatography:
Compound 21
0.095 g of diastereoisomer B (93 %,
MH+: 415).
Compound 21 (diastereoisomer B)
C. Analytical methods
The mass of the compounds was recorded with LCMS (liquid chromatography mass
spectrometry). Three methods were used which are described below. The data are
gathered in Table 1 below.
LCMS-method 1
LCMS analysis was carried out (electrospray ionization in positive mode, scanning
mode from 100 to 900 amu) on a Kromasil CIS column (mterchim, Montlucon, FR; 5
um, 4.6 x 150 mm) with a flow rate of 1 ml/minute. Two mobile phases (mobile phase
A: 30% 6.5mM ammonium acetate + 40% acetonitrile + 30% formic acid (2ml/l);
mobile phase B: 100% acetonitrile) were employed to run a gradient condition from
100 % A for 1 minute to 100% B in 4 minutes, 100% B for 5 minutes to 100 % A in 3
minutes, and reequilibrate with 100 % A for 2 minutes.
LCMS-method 2
LCMS analysis was carried out (electrospray ionization in both positive and negative
(pulsed) mode scanning from 100 to 1000 amu) on a Kromasil CIS column (Interchim,
Monttu$on, FR; 3.5 um, 4.6 x 100 mm) with a flow rate of 0.8 ml/minute. Two mobile
phases (mobile phase A: 35% 6.5mM ammonium acetate + 30% acetonitrile + 35%
formic acid (2ml/l); mobile phase B: 100% acetonitrile) were employed to run a
gradient condition from 100 % A for 1 minute to 100% B in 4 minutes, 100% B at a
flow rate of 1.2 mVrninute for 4 minutes to 100 % A at O.Sml/minute in 3 minutes, and
reequilibrate with 100 % A for 1.5 minute.
LCMS-method 3
LCMS analysis was carried out (electrospray ionization in both positive and negative
(pulsed) mode scanning from 100 to 1000 amu) on a Sunfire CIS column (Waters,
Millford USA; 3.5 um, 4.6 x 100 mm) with a flow rate of 0.8 ml/minute. Two mobile
phases (mobile phase A: 35% 6.5mM ammonium acetate + 30% acetonitrile + 35%
formic acid (2ml/l); mobile phase B: 100% acetonitrile) were employed to run a
gradient condition from 100 % A for 1 minute to 100% B in 4 minutes, 100% B at a
flow rate of 1.2 ml/minute for 4 minutes to 100 % A at O.Sml/minute in 3 minutes, and
reequilibrate with 100 % A for 1.5 minute.
LCMS-metfaod4
LCMS analysis was carried out (electrospray ionization in both positive and negative
(pulsed) mode scanning from 100 to 1000 amu) on a Sunfire CIS column (Waters,
Millford USA; 3.5 pm, 4.6 x 100 mm) with a flow rate of 0.8 ml/minute. Two mobile
phases (mobile phase A: 25% 6.5mM ammonium acetate + 50% acetonitrile + 25%
formic acid (2ml/l); mobile phase B: 100% acetonitrile) were employed to run a
gradient condition from 100 % A for 1 minute to 100% B in 4 minutes, 100% B at a
flow rate of 1.2 ml/minute for 4 minutes to 100 % A at 0.8 ml/minute in 3 minutes, and
reequilibrate with 100 % A for 1.5 minute.
D. 1. In-vitro method for testing compounds against M. tuberculosis.
Flat-bottom, sterile 96-well plastic microtiter plates were filled with 100 ul of
Middlebrook (Ix) broth medium. Subsequently, stock solutions (10 x final test
concentration) of compounds were added in 25 ul volumes to a series of duplicate wells
in column 2 so as to allow evaluation of their effects on bacterial growth. Serial fivefold
dilutions were made directly in the microtiter plates from column 2 to 11 using a
customised robot system (Zymark Corp., Hopkinton, MA). Pipette tips were changed
after every 3 dilutions to minimize pipetting errors with high hydrophobic compounds.
Untreated control samples with (column 1) and without (column 12) inoculum were
included hi each microtiter plate. Approximately 5000 CFU per well of
Mycobacterium tuberculosis (strain H37RV), hi a volume of 100 ul in Middlebrook
(Ix) broth medium, was added to the rows A to H, except column 12. The same volume
of broth medium without inoculum was added to column 12 in row A to H. The
cultures were incubated at 37°C for 7 days in a humidified atmosphere (incubator with
open air valve and continuous ventilation). One day before the end of incubation, 6
days after inoculation, Resazurin (1:5) was added to all wells in a volume of 20 ul and
plates were incubated for another 24 hours at 37°C. On day 7 the bacterial growth was
quantitated fluorometrically.
The fluorescence was read hi a computer-controlled fluorometer (Spectramax Gemini
EM, Molecular Devices) at an excitation wavelength of 530 nm and an emission
wavelength of 590 nm. The percentage growth inhibition achieved by the compounds
was calculated according to standard methods, and MIC data (representing IC90's
expressed in microgram/ml) were calculated.
1 against strain D.2. In-vitro method for testing compounds for anti-t M.
Smegmatis ATCC607.
Flat-bottom, sterile 96-well plastic microtiter plates were filled with 180 ul of sterile
deionized water, supplemented with 0.25 % BSA. Subsequently, stock solutions (7.8 x
final test concentration) of compounds were added in 45 ul volumes to a series of
duplicate wells in column 2 so as to allow evaluation of their effects on bacterial
growth. Serial five-fold dilutions (45 ul in 180 ul) were made directly in the microtiter
plates from column 2 to 11 using a customised robot system (Zymark Corp.,
Hopkinton, MA). Pipette tips were changed after every 3 dilutions to minimize
pipetting errors with high hydrophobic compounds. Untreated control samples with
(column 1) and without (column 12) inoculum were included in each microtiter plate.
Approximately 250 CPU per well of bacteria inoculum, in a volume of 100 ul in 2.8x
Mueller-Hinton broth medium, was added to the rows A to H, except column 12. The
same volume of broth medium without inoculum was added to column 12 in row A to
H. The cultures were incubated at 37°C for 48 hours in a humidified 5% CO2
atmosphere (incubator with open air valve and continuous ventilation). At the end of
incubation, two days after inoculation., the bacterial growth was quantitated
fluorometrically. Therefore Alatnar Blue (lOx) was added to all wells in a volume of 20
ul and plates were incubated for another 2 hours at 50°C.
The fluorescence was read in a computer-controlled fluorometer (Cytofluor, Biosearch)
at an excitation wavelength of 530 run and an emission wavelength of 590 nm (gain
30). The % growth inhibition achieved by the compounds was calculated according to
standard methods. The pICso was defined as the 50 % inhibitory concentration for
bacterial growth. The results are shown in Table 2.(Table Removed)




We Claim
1. A compound of formula
(Formula Removed)
the pharroaceutically acceptable add or base addition salts thereof, the quaternary
amines thereof the stereochemically isomeric forms thereof, the tautomeric forms
thereof and the N-oxide forms thereof; wherein:
R1 is hydrogen, halo, haloalkyl, cyano, hydroxy, Ar, Het, alkyl, aUcyloxy,
alkylthio, alkyioxyalkyl, alkylthioalkyl, Ar-alkyl or di(Ar)alkyl;
p isanintegerequal to l,2,3or4;
R2 is hydrogen, hydroxy, thio, alkyloxy, alkyloxyalkyloxy, alkylthio, mono
or di(alkyl)amino or a radical of formula wherein Y is CH2,
O,S,NH or N-alkyl;
R3 is alkyl, Ar, Ar-alkyl, Het or Het-alkyl;
R4 is hydrogen, alkyl or benzyl;
R5 is hydrogen, halo, haloalkyl, hydroxy, Ar, alkyl, aJiyloxy, alkylthio,
aDcyioxyalkyl, alkyltbioalkyl, Ar-alkyl or di(Ar)alkyl; or two vicinal R5 radicals may be taken together to form together with the phenyl ring to
which they are attached a naphthyl;

r is an integer equal to 1,2,3,4 or 5; and
R6 is hydrogen alkyl, Ar or Het;
R7 is hydrogen or alkyl;
R8 is oxo; ar
R7 and R8 together form the radical -CH=CS-N=;
Z isCH2or C(=O);
alkyl is a straight car branched saturated hydrocarbon radical having
carbon atoms; or is a cyclic saturated hydrocarbon radical having from 3 to 6 carbon atoms; or is a a cyclic saturated hydrocarbon radical havingfrom3 to 6 carbon atoms attached to a straight or banchedsatorated hydrocarbon radical having from 1 to 6 carbon atoms; wherein each carbon atom can be optionally substituted with halo, hydroxy, alkyloxyoroxo; Ar is a hornocycle selected ftom the group of phenyl, naphtbyl, acenapbfhyl,
tetrahydronaphihyl, each optionally substituted with 1,2 or 3 substitoente, each sobstituent independently selected ftom the group of hydroxy, halo, cyaoo, nitro, amino, mono- or dialkylamino, alkyl, haloaliyl, alkyloxy, haloalkyloxy, carboxyl, aliyloxycarbonyl, aminocarbanyl, morphoKnyl and mono- or dialkylaminocarbonyl; Het is anMnocyclic heterocycle selected from the group of N-phenoxypiperidinyl
pyrrolyl, pyrazolyl, imidazolyl, furanyl, thienyl oxazolyl, isoxazlyl, thiazolyl, isothiazolyl, pyridmyl, pyramidmyi, pyiazinyl and pyridasdnyl; or a bicyclic heterocycle selected from the groin) of quinolinyl, qmnoxalmyl, indolyl, benzimidazolyl, benzoxazolyl, benzisoxazolyL benzothiazoryl, benzisothiazolyl, benzofbranyl, benzomienyl, 2,3-dihydrobenzo[l,4]dioxinyl or benzo[l,3]dioxolyl; each monocyclic and bicyclic heterocycle may optionally besubstitued on a crabon atom with 1,2 or 3 substitoents selected from the group of halo, hydroxy, alkyl or alkyloxy; halo is a substituent selected from the group of fluoro, chloro, bromo and iodo and haloalkyl is a straight orbranched saturated hydrocarbon radical having from 1 to
6 carbon atoms or a cyclic saturated hydrocarbon radical having from 3 to 6 carbon atoms, wherein one or more carbon atoms are substituted with one or more halo-atoms.
2. The compound as claimed in claim 1 wherein Z is CH2.
3. The compound as claimed in any one of the preceding claims wherein R5 is hydrogen, halo, haloalkyl, hydroxy, Ar, alkyl, alkyloxy, alkylthio, alkyloxyalkyl, alkyithioalkyl, Ar-alkyl or di(Ar)alkyl.
4. The compound as claimed in claim 1 or 2 wherein

R1 is hydrogen, halo, cyano, At, Het, alkyl, and alkyloxy;
p is an integer equal to 1,2,3 or 4;
R2 is hydrogen, hydroxy, alkyloxy, alkyloxyalkyloxy, alkylthio or a radical
of formula . (Formula Removed) wherein Y is O;
R3 isalkyl, Ar, Ar-alkyl or Het;
R4 is hydrogen, alkyl or benzyl;
R5 is hydrogen, halo or alkyl; or
two vicinal R5 radicals may be taken together to form together with the phenyl ring to which they are attached a naphthyl;
r is an integer equal to 1; and
R6 is hydrogen;
R7 is hydrogen or alkyl;
R8 is oxo; or
R7 and R8 together form the radical -CH=CH-N=;
alkyl is a straight or branched saturated hydrocarbon radical having from 1 to 6
carbon atoms; or is a cyclic saturated hydrocarbon radical having from 3 to 6 carbon atoms; or is a acyclic saturated hydrocarbon radical having from 3 to 6 carbon atoms attached to a straight or branched saturated hydrocarbon radical having from 1 to 6 carbon atoms; wherein each carbon atom can be optionally substituted with halo or hydroxy;
Ar is a homocycle selected from the group of phenyl, naphthyl, acenaphthyl,
tetrahydronaphthyl, each optionally substituted with 1,2 or 3 substituents, each substituent independently selected from the group of halo, haloaikyl, cyano, alkyloxy and morpholinyl; Het is a monocyclic heterocycle selected from the group of N-phenoxypiperidinyl, furanyl, thienyl, pyridinyl, pyrimidinyl; or a bicyclic heterocycle selected from the group of benzofhienyl, 2,3-dihydrobenzo[l,4]dioxinyl or benzo[l,3]- dioxolyl; each monocyclic and bicyclic heterocycle may optionally be substituted on a carbon atom with 1,2 or 3 alkyl substituents; and halo is a substituent selected from the group of fiuoro, chloro and bromo.
5. The compound as claimed in any one of the preceding claims wherein the compound is a compound of formula (la) and wherein R1 is hydrogen, halo, Ar, Het; alkyl or alkyloxy; p =1; R2 is hydrogen, alkyloxy or alkylthio; R3 is naphthyl, phenyl or Het; each optionally substituted with 1 or 2 substituents selected from the group of halo and haloaikyl; R4 is hydrogen or alkyl; R5 is hydrogen, alkyl or halo; r is equal to 1 and R6 is hydrogen.
6. The compound as claimed in any one of claims 1, 3, 4 or 5, wherein the compound is a compound according to formula (la) wherein R1 is hydrogen, halo, alkyl, or Het; R2 is alkyloxy; R3 is naphthyl, phenyl or Het; each optionally substituted with halo; R4 is alkyl; R5 is hydrogen or halo; R6 is hydrogen; Z is CH2 or C(=O).

7. The compound which is degraded in vivo to yield a compound according to any one of the
preceding claims.
8. The compound as claimed in any one of the preceding claims for use as a medicine.
9 A process for preparing a compound as claimed in claim 1, characterized by
a) reacting an intermediate of formula (II-a) with paraformaldehyde in a suitable solvent
(Formula Removed)
With R1 to R8, p and r as defined in claim 1;
or if desired, converting the compounds of formula (Ia) into a therapeutically active non-toxic acid addition salt by treatment with an acid, or into a therapeutically active non-toxic base addition salt by treatment with a base, or conversely, converting the acid addition salt form into the free base by treatment with alkali, or converting the base addition salt into the free acid by treatment with acid, and, if desired, preparing stereochemically isomeric forms, quaternary amines, tautomeric forms or N-oxide forms thereof.


Documents:

4215-delnp-2006-Abstract-(06-04-2011).pdf

4215-delnp-2006-Claims-(06-04-2011).pdf

4215-delnp-2006-claims.pdf

4215-DELNP-2006-Correspondence Others-(14-06-2011).pdf

4215-delnp-2006-Correspondence-Others-(06-04-2011).pdf

4215-delnp-2006-Correspondence-Others-(15-04-2011).pdf

4215-delnp-2006-Correspondence-Others-(20-04-2011).pdf

4215-delnp-2006-correspondence-others-1.pdf

4215-delnp-2006-correspondence-others.pdf

4215-delnp-2006-description (complete).pdf

4215-delnp-2006-form-1.pdf

4215-delnp-2006-form-18.pdf

4215-delnp-2006-form-2.pdf

4215-delnp-2006-Form-3-(06-04-2011).pdf

4215-DELNP-2006-Form-3-(14-06-2011).pdf

4215-delnp-2006-form-3.pdf

4215-delnp-2006-form-5.pdf

4215-delnp-2006-GPA-(06-04-2011).pdf

4215-delnp-2006-gpa.pdf

4215-delnp-2006-pct-210.pdf

4215-delnp-2006-pct-237.pdf

4215-delnp-2006-pct-304.pdf

4215-delnp-2006-Petition 137-(06-04-2011).pdf

4215.doc


Patent Number 248028
Indian Patent Application Number 4215/DELNP/2006
PG Journal Number 24/2011
Publication Date 17-Jun-2011
Grant Date 09-Jun-2011
Date of Filing 21-Jul-2006
Name of Patentee JANSSEN PHARMACEUTICA N.V.
Applicant Address TURNHOUTSEWEG 30, 2340 BEERSE, BELGIUM.
Inventors:
# Inventor's Name Inventor's Address
1 GUILLEMONT, JEROME EMILE GEORGES C/O JOHNSON & JOHNSON PHARMACEUTICAL, RESEARCH AND DEVELOPMENT, A DIVISION OF JANSSEN-CILAG, CAMPUS DE MAIGREMONT, BP 615, 27106 VAL DE REUIL CEDEX, FRANCE
2 PASQUIER, ELISABETH THERESE JEANNE JOHNSON & JOHNSON PHARMACEUTICAL, RESEARCH AND DEVELOPMENT, A DIVISION OF JANSSEN-CILAG, CAMPUS DE MAIGREMONT, BP 615, 27106 VAL DE REUIL CEDEX, FRANCE
PCT International Classification Number C07D 413/06
PCT International Application Number PCT/EP2005/050267
PCT International Filing date 2005-01-21
PCT Conventions:
# PCT Application Number Date of Convention Priority Country
1 60/538768 2004-01-23 U.S.A.