Anti-inflammatory agents
The invention relates to aryl substituted 3-aminolactam derivatives and their use in
preventing or treating inflammatory diseases.
Inflammation is an important component of physiological host defence. Increasingly,
however, it is clear that temporally or spatially inappropriate inflammatory responses
play a part in a wide range of diseases, including those with an obvious leukocyte
component (such as autoimmune diseases, asthma or atherosclerosis) but also in
diseases that have not traditionally been considered to involve leukocytes (such as
osteoporosis or Alzheimer's disease).
The chemokines are a large family of signalling molecules with homology to
interleukin-8 which have been implicated in regulating leukocyte trafficking both in
physiological and pathological conditions. With more than fifty ligands and twenty
receptors involved in chemokine signalling, the system has the requisite information
density to address leukocytes through the complex immune regulatory processes from
the bone marrow, to the periphery, then back through secondary lymphoid organs.
However, this complexity of the chemokine system has at first hindered
pharmacological approaches to modulating inflammatory responses through
chemokine receptor blockade. It has proved difficult to determine which chemokine
receptor(s) should be inhibited to produce therapeutic benefit in a given inflammatory
disease.
More recently, a family of agents which block signalling by a wide range of
chemokines simultaneously has been described (see Reckless et al, Biochem J. (1999)
340: 803-81 1). The first such agent, a peptide termed "Peptide 3", was found to
inhibit leukocyte migration induced by 5 different chemokines, while leaving
migration in response to other chemoattractants (such as fMLP or TGF-beta)
unaltered. This peptide, and its analogs such as NR58-3.14.3 (i.e. c(DCys-DGln-DIle-
DT -DLys-DGln-DLys-DPro-DAs r-DLeu-DCys)-NH2 [SEQ ID NO: 1]), are
collectively termed "Broad Spectrum Chemokine Inhibitors" (BSCIs). Grainger et al.
(2003, Biochem. Pharm. 65: 1027-1034) have subsequently shown BSCIs to have
potentially useful anti-inflammatory activity in a range of animal models of diseases.
Interestingly, simultaneous blockade of multiple chemokines is not apparently
1
P66854. OO Spec as Hied 8.06.1
associated with acute or chronic toxicity, suggesting this approach may be a useful
strategy for developing new anti-inflammatory medications with similar benefits to
steroids but with reduced side-effects. This beneficial risk:benefit profile most likely
results from the unexpected mechanism of action of these compounds (see
International Patent Appl. No. PCT/GB20 10/0003 54 in the name of Cambridge
Enterprise Limited filed 28 February 2010, and International Patent Appl. No.
PCT/GB20 10/000342 in the name of Cambridge Enterprise Limited filed 26 February
2010).
However, peptides and peptoid derivatives such as NR58-3.14.3, may not be optimal
for use in vivo. They are quite expensive to synthesise and have relatively
unfavourable pharmacokinetic and pharmacodynamic properties. For example, NR58-
3.14.3 is not orally bioavailable and is cleared from blood plasma with a half-life
period of less than 30 minutes after intravenous injection.
Two parallel strategies have been adopted to identify novel preparations that retain the
anti-inflammatory properties of peptide 3 and NR58-3. 14.3, but have improved
characteristics for use as pharmaceuticals. Firstly, a series of peptide analogs have
been developed, some of which have longer plasma half-lives than NR58-3.14.3 and
which are considerably cheaper to synthesise (see for example WO2009/0 17620).
Secondly, a detailed structure: activity analysis of the peptides has been carried out to
identify the key pharmacophores and design small non-peptidic structures which retain
the beneficial properties of the original peptide.
This second approach yielded several structurally distinct series of compounds that
retained the anti-inflammatory properties of the peptides, including 16-amino and 16-
aminoalkyl derivatives of the alkaloid yohimbine, as well as a range of N-substituted
3-aminoglutarimides, identified from a small combinatorial library (see Fox et ai,
2002, J Med Chem 45: 360-370; WO 99/12968 and WO 00/42071). All of these
compounds are broad-spectrum chemokine inhibitors that retain selectivity over nonchemokine
chemoattractants, and a number of them have been shown to block acute
inflammation in vivo.
The most potent and selective of the above-mentioned aminoglutarimides was (S)-3-
(undec- 10-enoyl)-aminoglutarimide (NR58,4), which inhibited chemokine-induced
2
P66854.WO01 .Spec as filed 8.06.
migration in vitro with an ED50 of 5nM. This compound was orders of magnitude
more potent than 3-aminoglutarimides with more complex acyl side chains (such as
benzoyl or tert-butyloxo (Boc) groups). As a result, subsequent studies of
aminoglutarimide and aminolactam BSCIs have focussed almost exclusively on
compounds with simple linear and branched alkyl side chains.
However, further studies revealed that the aminoglutarimide ring was susceptible to
enzymatic ring opening in serum. Consequently, for some applications (for example,
where the inflammation under treatment is chronic, such as in autoimmune diseases)
these compounds may not have optimal properties, and a more stable compound with
similar anti-inflammatory properties may be superior.
As an approach to identifying such stable analogs, various derivatives of (S)- 3-
(undec-lO-enoyl)-aminoglutarimide have been tested for their stability in serum. One
derivative, the 6-deoxo analog (S)-3-(undec-10-enoyl)-tetrahydropyridin-2-one, is
completely stable in human serum for at least 7 days at 37°C, but has considerably
reduced potency compared with the parental molecule.
One such family of stable, broad spectrum chemokine inhibitors (BSCIs) are the 3-
amino caprolactams, with a seven-membered monolactam ring (see, for example,
WO2005/053702 and WO2006/016152). However, further useful anti-inflammatory
compounds have also been generated from other 3-aminolactams with different ring
size (see for example WO2006/1 34385). Other modifications to the lactam ring,
including introduction of heteroatoms and bicyclolactam ring systems, also yield
compounds with BSCI activity (see, for example, WO2006/018609 and
WO2006/085096).
In general, these earlier studies have demonstrated that the BSCI activity is conferred
on the molecule by the cyclic "head group" (a 3-amino lactam or imide) and defined,
to an extent, the structural limitations for activity (for example, bulky substituents on
the ring nitrogen are detrimental for activity, but variations in ring size have little
impact). To be active as a BSCI, this "head group" must have an acyl "tail group"
attached. Compounds with a 3-amino group, either free or N-alkyl substituted, bearing
a positive charge at physiological pH are completely inactive as BSCIs. Previous
3
P6685 4. OO Spec as filed 8.06. 11
disclosures have shown that this "tail group" can be linked to the "head group"
through simple amide, sulfonamide, urea or carbamate linkers.
While the structure of the "head" group and linker are critical for BSCI activity, it has
been shown that a wide variety of "tail groups" can be selected with out affecting the
primary pharmacology of the compound, at least in vitro. As a result, modification of
the "tail group" has been extensively used to optimise the physical and pharmaceutical
properties of the compounds. Changes in the structure of the "tail group" can, for
example, change the primary route of metabolism or excretion, modify the
pharmacokinetics or oral bioavailability, and thus act as the primary determinant of the
ADME properties of a selected compound.
Although the universe of possible "tail groups" known to retain BSCI activity for
suitable aminolactam "head groups" is very large, some "tail groups" have been
described as preferred. In some cases, structural features of the "tail group" have been
identified which increase the potency of BSCI activity of the aminolactam compound.
The most obvious such example is the introduction of 2',2' disubstitution, with a
tetrahedral sp3 arrangement at the 2' carbon centre in the tail group (the so-called "key
carbon"), which confers a 0-fold increase in potency as a BSCI, at least in vitro,
compared to a related compound lacking 2'2'-disubstitution. For example, 2'2'-
dimethyldodecenanoyl-3-aminocaprolactam is 10-fold more potent as a BSCI in the
MCP-1 induced THP-1 cell migration than assay than dodecanoyl-3-
aminocaprolactam (as disclosed previously in WO2005/053702), or indeed any other
related compound with a linear alkyl "tail group". The increased potency for branched
alkyl "tail groups" is restricted to branching at the 2' position - 3'3'-
dimethyldodecanoyl-3-aminocaprolactam is no more potent than the linear alkyl
analogs.
In other cases, structural features of the "tail group" have been identified which are
associated with improved ADME properties. For example, the pivoyl "tail group" of
2'2'-dimethylpropanoyl-3-aminovalerolactam contributes to the unexpected, and
particularly favourable, pharmaceutical properties of this molecule (as disclosed
previously in WO2009/01 6390). In particular, the pivoyl group is resistant to
4
P66854.WOO1.Spec as filed 8.06.1 1
metabolism, and therefore contributes to the unusually prolonged biological half-life of
this compound.
In marked contrast, other possible "tail groups" have generally been less preferred.
For example, compounds with a planar (sp2) carbon centre at the 2' position (such as
dodec-2',3'-enoyl-3-aminocaprolactam) have markedly lower potency as BSCIs than
compounds with corresponding saturated alkyl "tail groups". Similarly, the data from
the original library of glutarimides suggested that aromatic rings at the 2-position were
also substantially less active (Fox et al, 2002, J Med Chem 45: 360-370). Taken
together, these two findings have led to the reasonable assumption that aminolactams
with aromatic "tail groups", such as benzoyl or substituted benzoyl, would not be
useful as BSCIs. As a result, previous disclosures of compounds with BSCI activity
have all excluded such aromatic "tail groups".
The present invention discloses a series of 3-aminolactam compounds with aromatic
"tail groups", as well as pharmaceutical compositions comprising the compounds, and
medical uses of the compounds and compositions such as for the treatment of
inflammatory diseases. Surprisingly, all of the compounds as set out below have
substantial BSCI activity (greater than either 2',3 '-unsaturated acyl 3-aminolactams or
benzoylaminoglutarimides).
In one aspect of the invention, there is provided according to the invention is a
compound of general formula (I), or a pharmaceutically acceptable salt thereof, for
in the treatment of an inflammatory disorder:
(I)
P66854.WO0 .Spec as filed 8.06.1
wherein
n is an integer from 1 to 4;
k is an integer from 0 to 5, representing the number of groups substituting C2, C3, C4,
C5 and/or C of the benzyl ring; and
X are linear or branched groups substituting the benzyl ring independently selected
from any one of the group consisting of: alkyl, haloalkyl, hydroxyalkyl, hydroxy,
alkoxy, amino, aminoalkyl, aminodialkyl, carboxy, and halogen;
with the proviso that:
when on the benzyl ring C2, C5 and C6 are unsubstituted, and C is unsubstituted or is
substituted with an hydroxy, alkoxy, amino, aminoalkyl, aminodialkyl, or halogen
group, then C3 is substituted with a halogen group; and
when on the benzyl ring C , C and C6 are unsubstituted, and C is unsubstituted or is
substituted with an alkyl, haloalkyl, hydroxyalkyl, hydroxy, alkoxy, amino,
aminoalkyl, aminodialkyl or carboxy group, then C4 is substituted with any one of the
group consisting of: alkyl group, haloalkyl group, hydroxyalkyl group, and carboxy
group.
The carbon atom at position 3 of the lactam ring is asymmetric and consequently, the
compounds according to the present invention have at least two possible enantiomeric
forms, that is, the "R" and "S" configurations. The present invention encompasses
each of the two enantiomeric forms and all combinations of these forms, including the
racemic "RS" mixtures. With a view to simplicity, when no specific configuration is
shown in the structural formula, it should be understood that each of the two
enantiomeric forms and their mixtures are represented.
Further provided is a compound of formula (G), or a pharmaceutically acceptable salt
thereof, for use in the treatment of an inflammatory disorder:
6
P66854.WO01 .Spec as filed 8.06. 11
')
wherein n, k and X are defined as for general formula (I) compounds above.
Compounds (I '), having the (S)-configuration at the stereocentre, are 5-100 fold more
potent as a BSCIs than the (R)-enantiomer of the same compound.
The invention also provides the use of a compound of general formula (I), or a
pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the
treatment of an inflammatory disorder:
(I)
wherein
n is an integer from 1 to 4;
k is an integer from 0 to 5, representing the number of groups substituting C2, C3, C4,
C5 and/or C6 of the benzyl ring; and
X are linear or branched groups substituting the benzyl ring independently selected
from any one of the group consisting of: alkyl, haloalkyl, hydroxyalkyl, hydroxy,
alkoxy, amino, aminoalkyl, aminodialkyl, carboxy, and halogen;
with the proviso that:
7
P66854.WO01 .Spec as filed 8.06.1 1
when on the benzyl ring C2, C5 and C are unsubstituted, and C4 is unsubstituted or is
substituted with an hydroxy, alkoxy, amino, aminoalkyl, aminodialkyl, or halogen
group, then C3 is substituted with a halogen group; and
when on the benzyl ring C2, C and C are unsubstituted, and C3 is unsubstituted or is
substituted with an alkyl, haloalkyl, hydroxyalkyl, hydroxy, alkoxy, amino,
aminoalkyl, aminodialkyl or carboxy group, then C4 is substituted with any one of the
group consisting of: alkyl group, haloalkyl group, hydroxyalkyl group, and carboxy
group.
Additionally provided is the use of a compound of formula ( ), or a pharmaceutically
acceptable salt thereof, in the manufacture of a medicament for the treatment of an
inflammatory disorder:
( )
wherein n, k and X are defined as for general formula (I) above.
Certain compounds have been found to be novel per se. Thus, in another aspect of the
invention, there is provided a compound of general formula (I):
(I)
wherein
8
P 685 4. OO Spec as filed 8.06.1 1
n is an integer from 1 to 4;
k is an integer from 1 to 5, representing the number of groups substituting C2, C , C4,
C5 and/or C6 of the benzyl ring;
when n is 1 or 2, X are linear or branched groups independently selected from any one
of the group consisting of: C or higher alkyl, haloalkyl with a C higher alkyl group,
hydroxyalkyl with a C7or higher alkyl group, 7 r gre ater alkoxy, aminoalkyl with a C4or
higher alkyl group, aminodialkyl with two C4 or higher alkyl groups, and carboxy; and
when n is 3 or 4, X are linear or branched groups independently selected from any one
of the group consisting of: alkyl, haloalkyl, hydroxyalkyl, hydroxy, alkoxy, amino,
aminoalkyl, aminodialkyl, carboxy, and halogen;
with the proviso that:
when n is 3 or 4 and on the benzyl ring C , C5 and C are unsubstituted, and C is
unsubstituted or is substituted with an hydroxy, alkoxy, amino, aminoalkyl,
aminodialkyl, or halogen group, then C3 is substituted with a halogen group;
when n is 3 or 4 and on the benzyl ring C2, C5 and C6 are unsubstituted, and C is
unsubstituted or is substituted with an alkyl, haloalkyl, hydroxyalkyl, hydroxy, alkoxy,
amino, aminoalkyl, aminodialkyl or carboxy group, then C4 is substituted with any one
of the group consisting of: alkyl group, haloalkyl group, hydroxyalkyl group, and
carboxy group; and
when n=3, X is other than 4'-methoxy, 3'-trifluoromethyl, or 3',4',5'-trimethoxy,
provided that the compound is none of the group consisting of: 3-(3'-
trifluoromethylbenzoylamino)-caprolactam, 3-(4'-methylbenzoylamino)-caprolactam,
3-(2'-aminobenzoylamino)-caprolactam, 3-(3',4'-dimethoxybenzoylamino)-
caprolactam, 3-(3',5'-di-iert-butyl -4'- hydroxybenzoylamino)-caprolactam, 3-(2',4'-
dimethoxybenzoylamino)-caprolactam, 3-(3'-methoxybenzoylamino)-caprolactam, 3-
(4'-trifluoromethylbenzoylamino)-caprolactam, 3-(2',3',4'-trimethoxybenzoylamino)-
caprolactam, 3-(2',6'-difluoromethylbenzoylamino)-caprolactam, 3-(2'-
9
P66854. OO1.Spec as filed 8.06. 11
fluoromethylbenzoylamino)-caprolactam, 3-(2 '-amino-3 '-hydroxy-4'-
methylbenzoylamino)-caprolactam, and 3-(3 ' ,5' -dimethylbenzoylamino)-caprolactam.
Also provided is a compound of formula ( ):
wherein n, k and X are defined herein for general formula (I),
provided that the compound is none of the group consisting of: (S)-3-(3'-trifluoromethylbenzoylamino)-
caprolactam, (S)-3-(4' -methylbenzoylamino)-caprolactam, (S)-
3-(4'-methoxybenzoylamino)-caprolactam, (S)-3-(2'-carboxybenzoylamino)-
caprolactam, and (S)-3-(3', 4,,5'-trimethoxybenzoylamino)-caprolactam.
WO2007/0038669 teaches diarylamine-containing compounds and their use as
moduclators of c-kit receptors. Various intermediate compounds are used in the
synthesis of the diarylamine-containing compounds. Any overlap of the intermediate
compounds is hereby disclaimed from the present invention.
EP0462949 together with the related publication Angelucci et al, 993, J Medicinal
Chemistry 36: 1512-1519 teach 7-membered 3-acylamino lactams as enhancers of
learning and memory. Any overlap of specific compounds [such as (R)- and (S)-3-(3'-
trifluoromethylbenzoylamino)-caprolactam, (S)-3-(4'-methoxybenzoylamino)-
caprolactam, and (S)-3-(3', 4',5'-trimethoxybenzoylamino)-caprolactam] or generic
compounds (notably where n=3 in compounds of generic formulae (I) and/or ( )
according to the present invention) mentioned in these documents is hereby disclaimed
from the present invention.
JP03206042 discloses the preparation of 5,6,7,8-tetrahydo-4H-thiazolo[5,4-b]azepine
derivatives with potassium channel activation activity, for use as antihypertensives.
10
P66854.WO0l .Spec as filed 8.06.1 1
Various intermediate compounds (notably where n=3 in compounds of generic
formulae (I) and/or ( ) according to the present invention) are used in the synthesis of
the derivatives. Any overlap of the intermediate compounds is hereby disclaimed from
the present invention.
The prior art also discloses specific compounds, for example:
3-(4'-methylbenzoylamino)-caprolactam is disclosed in Uchikawa et al. ( 1996)
Chemical & Pharmaceutical Bulletin 44: 2070-2077;
3-(2'-aminobenzoylamino)-caprolactam is disclosed in Uchikawa et al. ( 1994) J
Heterocyclic Chemistry 31: 877-887;
- (S)-3-(2'-carboxybenzoylamino)-caprolactam is disclosed in Belyaev (1995)
Tetrahedron Letters 36: 439-440 ;
3-(3'-trifluoromethylbenzoylamino)-caprolactam (in both (S)- and (R)- forms)
and (S)-3-(3', 4',5'-trimethoxybenzoylamino)-caprolactam are disclosed in
EP462949A1; 3-(3'-trifluoromethylbenzoylamino)-caprolactam is also
disclosed in EP351856A2;
- 3-(3',4'-dimethoxybenzoylamino)-caprolactam, 3-(3',5'-di-ter t-butyl -4'-
hydroxybenzoylamino)-caprolactam, 3-(2',4'-dimethoxybenzoylamino)-
caprolactam, 3-(3'-methoxybenzoylamino)-caprolactam, 3-(4'-
trifluoromethylbenzoylamino)-caprolactam, 3-(2',3',4'-
trimethoxybenzoylamino)-caprolactam, 3-(2',6'-difluoromethylbenzoylamino)-
caprolactam, and 3-(2'-fluoromethylbenzoylamino)-caprolactam are disclosed
in JP03206042A;
3-(2'-amino-3'-hydroxy-4'-methylbenzoylamino)-caprolactam is disclosed in
Kameda et al. (1968) Chemical & Pharmaceutical Bulletin 16: 480-485; and
- 3-(3'5'-dimethylbenzoylamino)-caprolactam is disclosed in the Aurora
Screening Library Catalogue published on 10 March 2010 (Order No.
07.167.701; CHEMCATS Acc. NO. 001 5557046).
11
P66854.WO01.Spec as filed 8.06.1 1
However, none of the above prior art compounds have been shown to have BSCI
activity, or to be useful for the treatment of inflammatory diseases. As a result,
compounds disclosed in the prior art documents mentioned herein in no way teach or
suggest our unexpected finding that the class of aryl-substituted aminolactams and
analogs as defined herein have useful BSCI activity, and the prior art compounds are
hereby disclaimed.
In another aspect of the invention, there is provided a pharmaceutical composition
comprising, as active ingredient, a compound per se as defined above, or a
pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable
excipient and/or carrier.
By pharmaceutically acceptable salt is meant in particular the addition salts of
inorganic acids such as hydrochloride, hydrobromide, hydroiodide, sulphate,
phosphate, diphosphate and nitrate or of organic acids such as acetate, maleate,
fumarate, tartrate, succinate, citrate, lactate, methanesulphonate, p-toluenesulphonate,
palmoate and stearate. Also within the scope of the present invention, when they can
be used, are the salts formed from bases such as sodium or potassium hydroxide. For
other examples of pharmaceutically acceptable salts, reference can be made to "Salt
selection for basic drugs" (1986) Int. J . Pharm. 33: 201-217.
The pharmaceutical composition can be in the form of a solid, for example powders,
granules, tablets, gelatin capsules, liposomes or suppositories. Appropriate solid
supports can be, for example, calcium phosphate, magnesium stearate, talc, sugars,
lactose, dextrin, starch, gelatin, cellulose, methyl cellulose, sodium carboxymethyl
cellulose, polyvinylpyrrolidine and wax. Other appropriate pharmaceutically
acceptable excipients and/or carriers will be known to those skilled in the art.
The pharmaceutical compositions according to the invention can also be presented in
liquid form, for example, solutions, emulsions, suspensions or syrups. Appropriate
liquid supports can be, for example, water, organic solvents such as glycerol or
glycols, as well as their mixtures, in varying proportions, in water.
Exemplar compounds according to general formula (I) and formula ( ) for medical
uses according to the invention may be selected from the group consisting of:
12
P668S4.WO01 .Spec as filed 8.06. 11
(S)-3 -(4'-methylbenzoylamino)-caprolactam, and
(S)-3-(3',5'-dimethylbenzoylamino)-caprolactam,
and pharmaceutically acceptable salts thereof.
Exemplar per se compounds of the invention according to general formula (G ), and/or
exemplar compounds according to general formula (G) and formula (P) for medical
uses according to the invention, may be selected from the group consisting of:
(5)-3-Fluoro-N-(2-oxopiperidin-3-yl)benzamide,
(5)-2-Fluoro-N-(2-oxopiperidin-3-yl)benzamide,
(5)-4-Fluoro-N-(2-oxopiperidin-3-yl)benzamide,
(5)-N-(2-Oxopiperidin-3-yl)-4-(trifluoromethyl)benzamide,
(5}-N-(2-Oxopiperidin-3-yl)-3-(trifluoromethyl)benzamide,
(5)-N-(2-Oxopiperidin-3-yl)-2-(trifluoromethyl)benzamide,
(S)-2,3-difluoro-N-(2-oxopiperidin-3-yl)benzamide,
(5}-2,4-difluoro-N-(2-oxopiperidin-3-yl)benzamide,
(5)-2,5-difluoro-N-(2-oxopiperidin-3-yl)benzamide,
(5)-2,6-difluoro-N-(2-oxopiperidin-3-yl)benzamide,
(5)-3,4-difluoro-N-(2-oxopiperidin-3-yl)benzamide,
(5)-3,5-difluoro-N-(2-oxopiperidin-3-yl)benzamide,
(S)-3-(3'-Butylbenzoylamino)-azepan-2-one,
(S)-3-(4'-Ethylbenzoylamino)-tetrahydropyridin-2-one,
(S)-3-(4'-Butylbenzoylamino)-tetrahydropyridin-2-one,
(S)-3-(4'-tert-Butylbenzoylamino)-tetrahydropyridin-2-one, and
(S)-3-(4'-Hexylbenzoylamino)-tetrahydropyridin-2-one,
and pharmaceutically acceptable salts thereof.
The compound (5)-4-Fluoro-N-(2-oxopiperidin-3-yl)benzamide is also known as (S)-
3-(4'-fluorobenzoylamino)-tetrahydropyridin-2-one ( see Example 3 below).
13
P668 5 4 .WOO Spec as filed 8.06.1
Exemplar per se compounds of the invention according to general formula (I) or (P),
and/or exemplar compounds according to general formula (I) and formula (P) for
medical uses according to the invention, may be selected from the group consisting of:
(S)-3-(4'-Ethylbenzoylamino)-azepan -2-one,
(S)-3-(4'-Butylbenzoylamino)-azepan -2-one,
(S)-3-(4'-tert-Butylbenzoylamino)-azepan -2-one,
(S)-3-(4'-Hexylbenzoylamino)-azepan -2-one,
(S)-3-(4'-Octylbenzoylamino)-azepan -2-one, and
(S)-3-(4'-Octylbenzoylamino)-tetrahydropyridin -2-one,
and pharmaceutically acceptable salts thereof.
Exemplar compounds according to general formula (I) for medical uses according to
the invention, may be selected from the group consisting of:
(R)-3-(4'-Butylbenzoylamino)-tetrahydropyridin -2-one,
(R)-3-(4'-tert-Butylbenzoylamino)-tetrahydropyridin -2-one, and
(R)-3-(4'-Hexylbenzoylamino)-tetrahydropyridin -2-one,
and pharmaceutically acceptable salts thereof.
Exemplar per se compounds of the invention according to general formula (I), and/or
exemplar compounds according to general formula (I) for medical uses according to
the invention, may be (R)-3-(4'-Octylbenzoylamino)-tetrahydropyridin -2-one or a
pharmaceutically acceptable salt thereof.
The compounds (R)-3-(4'-Butylbenzoylamino)-tetrahydropyridin -2-one,
(R)-3-(4'-½rt-Butylbenzoylamino)-tetrahydropyridin -2-one, (R)-3-(4'-Hexylbenzoylamino)-
tetrahydropyridin -2-one, and (R)-3-(4'-Octylbenzoylamino)-tetrahydropyridin-
2-one, and pharmaceutical salts of each, are a further aspect of the invention.
According to the invention, inflammatory disorders (which term is used herein
interchangeably with "inflammatory disease") intended to be prevented or treated by
the compounds of formula (I) or (P), or pharmaceutically acceptable salts thereof or
14
P66854.WO01 .Spec as filed 8.06. 1
pharmaceutical compositions or medicaments containing them as active ingredients,
include notably:
autoimmune diseases, for example such as multiple sclerosis, rheumatoid arthritis,
lupus, irritable bowel syndrome, Crohn's disease;
- vascular disorders including stroke, coronary artery diseases, myocardial
infarction, unstable angina pectoris, atherosclerosis or vasculitis, e. g., Behcet's
syndrome, giant cell arteritis, polymyalgia rheumatica, Wegener's granulomatosis,
Churg-Strauss syndrome vasculitis, Henoch-Schonlein purpura and Kawasaki
disease;
- asthma, and related respiratory disorders such as allergic rhinitis and COPD;
organ transplant rejection and/or delayed graft or organ function, e.g. in renal
transplant patients;
- psoriasis;
skin wounds and other fibrotic disorders including hypertrophic scarring (keloid
formation), adhesion formations following general or gynaecological surgery, lung
fibrosis, liver fibrosis (including alcoholic liver disease) or kidney fibrosis,
whether idiopathic or as a consequence of an underlying disease such as diabetes
(diabetic nephropathy); or
- allergies.
The inflammatory disorder may be selected from the group consisting of autoimmune
diseases, asthma, rheumatoid arthritis, a disorder characterised by an elevated TNF-a
level, psoriasis, allergies, multiple sclerosis, fibrosis (including diabetic nephropathy),
and formation of adhesions.
The above clinical indications fall under the general definition of inflammatory
disorders or disorders characterized by elevated TNFo levels.
15
P66854.WOO 1 Spec as filed 8.06. 11
In one aspect of the invention, merely in order to circumvent any potentially
conflicting prior art (for example as noted above), the term inflammatory disorder may
exclude cognitive disorders such as Alzheimer's disease and/or memory loss.
Compounds of formula (I) or ( ) are particularly useful for local delivery, and also for
the preparation of medicaments for local delivery, including creams and ointments for
topical delivery, powders, aerosols or emulsions for inhaled delivery, and solutions or
emulsions for injection. Pharmaceutical compositions containing one or more
excipients suitable for such local delivery are therefore envisaged, and subsequently
claimed.
Also provided according to the invention is a method of treatment, amelioration or
prophylaxis of the symptoms of an inflammatory disease (including an adverse
inflammatory reaction to any agent) by the administration to a patient of an anti
inflammatory amount of a compound, pharmaceutical composition or medicament as
defined herein.
Administration of a compound, composition or medicament according to the invention
can be carried out by topical, oral, parenteral route, by intramuscular injection, etc.
The administration dose envisaged for a compound, composition or medicament
according to the invention is comprised between 0.1 mg and 10 g depending on the
formulation and route of administration used.
The invention further encompasses a library consisting of elements all of which have
structures according to the formula (I) or ( ), and hence which all have anti
inflammatory activity, useful for screening compounds for novel or improved
properties in a particular assay of anti-inflammatory activity.
The invention includes compounds, compositions and uses thereof as defined, wherein
the compound is in hydrated or solvated form. Unless specified otherwise, compounds
of the invention include tautomers, resolved enantiomers, resolved diastereomers,
racemic mixtures, solvates, metabolites, salts and prodrugs thereof, including
pharmaceutically acceptable salts and prodrugs.
In any of the compounds described above, n may be 2. Alternatively, n may be 3.
16
P66854.WO0 1 Spec as filed 8.06.1 1
X may be haloalkyl, for example trifluoromethyl.
An exemplar group of compounds per se and/or for medical use according to any
aspect of the invention is selected from among compounds according to formula (I) or
(G ) where X is halogen or haloakyl and where k is between 1 and 3. For example, X
may be fluoro or fluoroalkyl (such as trifluoromethyl) and k may be between 1 and 3.
Where permissible according to the formulae herein, the benzyl ring may be
monosubstituted with a group X as defined above (i.e. k = 1). For example, the benzyl
ring may be monosubstituted with an alkyl group (such as other than para-methyl or
other than Ci- alkyl), haloalkyl (such as trifluoromethyl, for example paratrifluoromethyl
[i.e. 4'-trifluoromethyl], ortho-trifluoromethyl [i.e. 2'-triflouromethyl]
or meta-trifluoromethyl [i.e. 3'-trifluoromethyl]). The benzyl ring may be
monosubstituted with an haloalkyl other than a Ci-6 haloalkyl. The benzyl ring may be
monosubstituted with halogen. The benzyl ring may be monosubstituted with orthocarboxy
[i.e. 2'-carboxy].
The single substitution group X may in particular be located in the eta (i.e. 3'-)
position on the benzyl ring.
In one aspect, the above features for k=l apply when n=2.
Where permissible according to the formulae herein, n may be 3 and the benzyl ring
may be monosubstituted with a group X as defined above (i.e. k = 1). For example, the
benzyl ring may be monosubstituted with an alkyl group other than a Ci- alkyl.
According to the invention, the compounds of general formula (I) or ( ) can be
prepared using the processes described hereafter.
DEFINITIONS
The term "about" refers to an interval around the considered value. As used in this
patent application, "about X" means an interval from X minus 10% of X to X plus
10% of X, and preferably an interval from X minus 5% of X to X plus 5% of X.
The use of a numerical range in this description is intended unambiguously to include
within the scope of the invention all individual integers within the range and all the
17
P66854.WO01 .Spec as filed 8.06.1 1
combinations of upper and lower limit numbers within the broadest scope of the given
range. Hence, for example, the range of 0.lmg to 1Og specified in respect of (inter
alia) a dose of a compound or composition of the invention to be used is intended to
include all doses between O.lmg and lOg and all sub-ranges of each combination of
upper and lower numbers, whether exemplified explicitly or not.
As used herein, the term "comprising" is to be read as meaning or encompassing both
comprising and consisting of. Consequently, where the invention relates to a
"pharmaceutical composition comprising as active ingredient" a compound, this
terminology is intended to cover both compositions in which other active ingredients
may be present and also compositions which consist only of one active ingredient as
defined.
The term "alkyl" or "alkyl group" as used herein refers to a saturated linear or
branched- chain monovalent hydrocarbon radical, for example of one to twenty carbon
atoms, one to twelve carbon atoms, one to six carbon atoms, one to four carbon atoms,
or as otherwise specified herein. Examples of alkyl groups include, but are not limited
to, methyl (Me, -CH ), ethyl (Et, -CH CH3), 1-propyl (n-Pr, n-propyl, -CH2CH2CH ),
2-propyl (i-Pr, i-propyl, -CH(CH )2), -butyl (n-Bu, n- butyl, -CH2CH2CH2CH3), 2-
methyl-1 -propyl (i-Bu, i-butyl, -CH2CH(CH3)2), 2-butyl (s- Bu, s-butyl, -
CH(CH3)CH2CH3), 2-methyl-2-propyl (t-Bu, t-butyl, -C(CH3)3), 1-pentyl (n- pentyl, -
CH2CH2CH CH CH ), 2-pentyl (-CH(CH3)CH2CH2CH ), 3-pentyl (- CH(CH2CH3)2)5
2-methyl-2-butyl (-C(CHs) 2CH CH3), 3-methyl-2-butyl (- CH(CH )CH(CH )2), 3-
methyl-1 -butyl (-CH2CH2CH(CH )2), 2-methyl-l -butyl (- CH2CH(CH )CH2CH ), 1-
hexyl (-CH2CH2CH2CH2CH2CH3), 2-hexyl (- CH(CH3)CH2CH2CH2CH ), 3-hexyl (-
CH(CH2CH3)(CH2CH CH3)), 2-methyl-2-pentyl (- C(CHs)2CH2CH CH ), 3-methyl-2-
pentyl (-CH(CH3)CH(CH3)CH2CH ), 4-methyl-2- pentyl (-CH(CH3)CH2CH(CH3)2),
3-methyl-3-pentyl (-C(CH3)(CH2CH )2), 2-methyl-3- pentyl (-
CH(CH2CH )CH(CH )2), 2,3-dimethyl-2-butyl (-C(CH )2CH(CH3)2), 3,3- dimethylsbutyl
(-CH(CH )C(CH ) , 1-heptyl, and 1-octyl.
The term "haloalkyl" or "haloalkyl group" as used herein refers to an alkyl group (as
defined above) except that one or more or all of the hydrogens of the alkyl group is
replaced by a halogen, which replacement can be at any site on the alkyl, including the
18
P66854.WO01 .Spec as filed 8.06. 11
end. Examples include, but are not limited to, CH2F, CHF2, CF , CH CH2F5 CH CHF ,
CH2CF3, CHFCF , CF2CF3, CH C1, CHC12, CC13, CH2CH2C1, CH2CHC12, CH2CCI3,
CHC1CC13, and CC12CC13.
The term "halogen" (which may be abbreviated to "halo") or "halogen group" as used
herein includes fluorine (F), bromine (Br), chlorine (CI), and iodine (I).
The term "hydroxy" or "hydroxy group" denotes the group "-OH".
The term "hydroxyalkyl" or "hydroxyalkyl group" as used herein refers to an alkyl
group (as defined above) except wherein one or more or all of the hydrogens of the
alkyl group is replaced by an hydroxy group, which replacement can be at any site on
the alkyl, including the end.
The term "alkoxy" or "alkoxy group" denotes an alkyl group as defined above attached
via a divalent oxygen atom to the rest of the molecule. Examples include but are not
limited to methoxy (-OCH 3), ethoxy, propoxy, isopropoxy, n-butoxy, sec-butoxy, tertbutoxy,
pentoxy, isopentoxy, neopentoxy, hexoxy, and 3-methylpentoxy.
The term "amino" or "amino group" denotes the group "-NH 2".
The term "aminoalkyl" or "aminoalkyl group" refers to an amino group in which one
of the hydrogen atoms has been replaced by an alkyl group as defined above.
The term "aminodialkyl" or "aminodialkyl group" refers to an amino group in which
both of the hydrogen atoms have been replaced by an alkyl group as defined above.
The alkyl groups attached to the nitrogen atom may be different or the same.
The term "carboxy" or "carboxy group" denotes the group "-C(0)OH".
The term "benzyl ring" (also known as a "phenyl group") refers to a 6 carbon aryl
group in compounds of general formulae (I) or ( ) shown above. For the purposes of
the general formulae of the present invention, numbering to locate the carbon atoms
C2-C within the benzyl ring is in a clockwise direction from Ci which is linked to the
3-aminolactam group. However, numbering of ring carbons with respect to one or
more substituent groups on the benzyl ring for specific compounds follows the IUPAC
rule that the second substituent in a clockwise or counter clockwise direction is
19
P66854.WO0l .Spec as filed 8.06.1 1
afforded the lower possible location number. Where two or more substituents are
present in a specific compound, the IUPAC rule is that they are listed in alphabetical
order. Location numbers on the ring are assigned according to the IUPAC rule to the
substituents so that they have the lowest possible number (starting from C which is
linked to the 3-aminolactam group), counting in either a clockwise or counter¬
clockwise direction.
As would be understood by a person skilled in the art, where there are fewer than 5
groups substituting the benzyl ring in compounds of general formulae (I) or (P), i.e.,
where k=0, 1 2, 3 or 4, the or each unsubstiruted position is occupied by a hydrogen
atom.
Unless otherwise defined, all the technical and scientific terms used here have the
same meaning as that usually understood by an ordinary specialist in the field to which
this invention belongs. Similarly, all the publications, patent applications, all the
patents and all other references mentioned here are incorporated by way of reference
(where legally permissible).
Preparation of the compounds of general formula (I) or (F)
All the compounds of general formula (I) or (F) can be prepared easily according to
general methods known to the person skilled in the art.
Typically, such compounds are made by coupling the "tail group" in the form of a
suitably activated acid (such as an acid chloride) with the appropriate 3-aminolactam.
Methods for the preparation of 3-aminolactams with 5,6,7 and 8 membered rings,
encompassing all the compounds claimed herein, have been extensively described in
the literature. For example, we have provided suitable methods for the preparation of
6-membered aminolactams from ornithine (see WO2009/0 16390) and 7-membered
aminolactams from lysine (see WO2005/053702), as well as methods for 5- and 8-
membered aminolactams (see WO2006/1 34385). We have described in particular
detail various synthesis routes to the 6-membered aminolactam, including processes
suitable for scaling up the manufacture to Kg quantities (WO2009/01 6390). Various
other methods for the synthesis of 3-aminolactams of various ring sizes have also been
described in the literature (see for example Pellegata et al., 1978, Synthesis 614-616
20
P66854 WOO Spec as filed 8.06. 11
and Boyle et ai, 79, J Org Chem 44:4841-4847), and any suitable method for the
preparation of the aminolactam "head group" may be employed in accordance with the
method of the present invention.
In the second step, the 3-aminolactam product is reacted with an appropriate acid
chloride, for example as previously described for 7-ring aminolactams (Fox et al,
2005, J Med Chem 48: 867-74). This reaction may be carried out, for example, in
chloroform or dichloromethane. The most preferred reaction solvent is
dichloromethane, and is preferably carried out in the presence of a base, for example
Na C0 3. The above reaction may be carried out at ambient temperature (about 25 °C)
or more generally at a temperature between 20 and 50 °C. The two reactions may be
carried out independently, with separation and purification of the 3-aminolactam
between the reactions, or alternatively, the reactions may be performed in a single
vessel without purification of the 3-aminolactam prior to its derivatisation with acid
chloride.
As noted previously (see WO2009/01 6390) care must be exercised during the
acylation reaction when preparing an enantiomerically pure compound, according to
formula (G ) by acylating an enantiomeriocally pure 3-aminolactam. In particular, the
base, such as sodium carbonate, must be added slowly continually monitoring the pH
of the reaction vessel to ensure that the pH of the reaction remains below pH 9.0
throughout. Excess basicity, for example due to rapid or excessive addition of sodium
carbonate, increases the racemisation of the 3-aminolactam and yields
enantiomerically impure product.
The following examples are presented in order to illustrate the above procedures and
should in no way be considered to limit the scope of the invention.
FIGURES
Fig. 1 shows the chemical structure of various examples of compounds according to
the inventions and reference examples; and
Fig. 2 . is a graph showing the results of a murine sub-lethal endotoxemia test. In the
graph, column A shows data from a control group (1% CMC lOml/kg p.o.), and
2 1
P668 4.WO01 .Spec as filed 8.06.1 1
column B shows data from a group treated with lmg/kg p.o. (5)-4-Fluoro-N-(2-
oxopiperidin-3-yl)benzamide (a compound according to one embodiment of the
present invention - see also Example 3 below) The y-axis shows levels of TNF-a in
pg/ml.
EXAMPLES
In the following further examples, -NMR and C-NMR spectra were recorded on a
Bruker Avance DRX 400 MHz fourier transform machine and F-NMR spectra were
recorded on a Bruker Avance DRX 300. Chemical shifts are given in ppm and
coupling constants, J, are given in Hz to the nearest 0.5. IR spectra were recorded on
an Avatar 320. HRMS data was gained via an Esquire 2000. [ C ] D values were recorded
on an optical activity AA 1000 polarimeter set at 598 nm (Sodium D line). The
samples were made using spectroscopic grade MeOH.
Reference Example 1: 3-(Benzoylamino)tetrahydropyridin-2-one:
3-aminotetrahydropyridin-2-one hydrochloride (10 mmol) and K2C0 (30 mmol) were
added to water (20 mL) and stirred. A solution of benzoyl chloride (10 mmol) in
CH C 12 (10 mL) was added and the reaction was stirred overnight at room temperature
in an inert atmosphere (using dinitrogen). The reaction was extracted with CH C 1 (3
50 mL), and the combined organic layers where then dried (Na2S04) and reduced in
vacuo to give a crude product which was recrystallised from CH2C 12 / petroleum ether
(bp 40-60 °C) to give the product (1.62 g, 74%):
Vmax cm 1 3250 (N-H, amide), 1664, 1633, 1538 (secondary CONH, lactam), 1605,
1578, 1486 (aromatic ring), 766, 715, 704, 690 (monosubstituted benzene ring).
NMR: d (400MHz, CDC1 ) 7.80 (2H, br d, J 7.0, ortho-H), 7.47-7.40 ( 1H, m,
para-U), 7.42-7.39 (1H, m, C6H5-CON H), 7.40-7.31 (2H, m, meta-W), 6.78 (1H, br s,
CONH-CH2), 4.41 (1H, dt, J 11.5, 5.5, CH-CO), 3.36-3.23 (2H, m, CH2NH), 2.59 (1H,
22
P66854. WOO Spec as Hied 8.06 . 1
dq, J 13.0, 4.5, NHCH-C H;), 1.94-1.81 (2H, m, lactam CH2), 1.64 (IH, tt, J 12.5, 8.0,
NHCH-CH^).
1 C NMR: C (100MHz, CDC1 ) 171.9 (lactam C=0), 167.4 (aryl C=0), 134.0 (ipso-
C), 131 .4 (ortho-C), 128.3 (meta-C), 127.0 (para-C), 50.8 (CH-CO), 41.5 (CH2NH),
27.0 (lactam CH2), 20.9 (lactam CH2).
HRMS (+ESI) Ci2H, N 0 2 + Na+: calcd 241.0947; found 241 .0950.
Reference Example 2 : 3-(Benzoylamino)azepan-2-one:
3-aminoazepan-2-one hydrochloride (10 mmol) and K2C0 3 (30 mmol) were added to
water (20 mL) and stirred. A solution of benzoyl chloride (10 mmol) in CH2CI2 (10
mL) was added and the reaction was stirred overnight at room temperature in an inert
atmosphere (using dinitrogen). The reaction was extracted with CH2C 12 (3 50 mL),
and the combined organic layers where then dried (Na2S0 4) and reduced in vacuo to
give a crude product which was recrystallised from ¾ 12 / petroleum ether (bp 40-
60 °C) to give the product ( 1.59 g, 68%):
m m 1 3244, 3202 (N-H, amide), 1660, 1642, 1536 (secondary CONH, lactam),
1601, 1578, 1536 (aromatic ring), 771, 707 (monosubstituted benzene ring).
NMR: H (400MHz, CDC13) 7.86-7.80 (2H, , ortho-H), 7.65 (IH, d, J 4.0, C6H5-
CONH), 7.52-7.45 ( 1H, m,para-H), 7.46-7.39 (2H, , meta-U), 6.1 1 ( 1H, br s,
CONH-CH2), 4.72 (IH, ddd, J 11.0, 5.5, 1.5, CH-CO), 3.49-3.39 (2H, , CH NH),
2.25 (IH, br d, J 13.5, lactam CH2), 2.08-1.98 (IH, m, lactam CH2), 1.94-1.82 (2H, ,
lactam CH2), 1.62-1.49 (IH, , lactam CH2), 1.49-1.36 (IH, m, lactam CH2) .
1 C NMR: C (100MHz, CDC1 ) 175.9 (lactam C=0), 166.4 (aryl C=0), 134.3 (ipso-
C), 131.7 (ortho-C), 128.7 (meta-C), 127.2 (para-C), 52.7 (CH-CO), 42.3 (CH -NH),
31.7 (lactam CH2) 29.0 (lactam CH ), 28.1 (lactam CH ) .
HRMS (+ESI) C|2H,4N20 2 + H+: calcd 233.1285; found 233.1283.
23
P66854. WOO Spec as Hied 8.06.1 1
Reference Example 3: 3-(4'-Methylbenzoylamino)tetrahydropyridin-2-one:
3-aminotetrahydropyridin-2-one hydrochloride (10 mmol), K2C0 3 (30 mmol) and 4-
methylbenzoyl chloride (10 mmol) were reacted according to the above procedure to
give the product ( .62 g, 78%):
ma cm 1 3306, 3203 (N-H, amide), 1669, 1649 (secondary CONH, lactam), 1612,
1489 (aromatic ring), 842, 810 (para-disubstituted benzene ring).
Ή NMR: d (400MHz, CDC13) 7.69 (2H, br d, J 8.0, ortho-H), 7.20 (2H, br d, J 8.0,
meta-H), 7.13 (IH, br d, J 4.0, CONH-CH ), 6.03 (IH, br s, C7H7-CONH), 4.41 ( 1H,
dt, J 11.0, 5.5, CH-CO), 3.41-3.30 (2H, , CH2NH), 2.72 (IH, dq, J 13.0, 4.5, lactam
CH2), 2.36 (3H, s, CH3), 2.05-1.90 (2H, , lactam CH2), 1.68-1.54 (IH, , lactam
CH ) .
1 C NMR: c (100MHz, CDC1 ) 172.3 (lactam C=0), 167.7 (aryl C=0), 142.1 (ipso-
C), 131.2 (para-C), 129.3 (aromatic-CH), 127.3 (aromatic-CH), 51.1 (CH-CO), 41.8
(CH2-NH), 27.4 (lactam CH2), 21.7 (CH3), 21.2 (lactam CH2) .
HRMS (+ESI) Ci H, N20 2 + Na+: calcd 255.1 104; found 255.1 104.
Reference Example 4 : 3-(4'-MethylbenzoyIaniino)azepan-2-one :
3-aminoazepan-2-one hydrochloride (10 mmol), 2C0 3 (30 mmol) and 4-
methylbenzoyl chloride (10 mmol) were reacted according to the above procedure to
give the product ( .63 g, 74%):
Vmax cm 1 3265, 3219 (N-H, amide), 1663, 1 47 (secondary CONH, lactam), 1607,
1570, 1505 (aromatic ring), 838, 823 (para-disubstituted benzene ring).
NMR: d (400MHz, CDC13) 7.72 (2H, br d, J 8.0, ortho-H), 7.58 (IH, br d, J 4.50,
CONH-CH2), 7.21 (2H, d, J 8.0, met - ), 5.98 (IH, br s, C7H7-CONH), 4.69 ( 1H,
ddd, J 1.0, 5.5, 2.0, CH-CO), 3.40-3.321 (2H, m, CH2NH), 2.37 (3H, s, CH ), 2.23
24
P66854.WO0l .Spec as filed 8.06.1 1
(1H, br d, lactam CH2), 2.08-1 .99 (1H, , lactam CH ), 1.96-1 .83 (2H, , lactam
CH2), 1.60-1.51 (2H, m, lactam CH2).
C NMR: c (100MHz, CDC1 ) 176.1 (lactam C=0), 66.3 (aryl C=0), 142.0 (ipso-
C), 13 1.5 (para-C), 129.3 (aromatic-CH), 127.2 (aromatic-CH), 52.6 (CH-CO), 42.2
(CH -NH), 31.7 (lactam CH2), 29.0 (lactam CH2), 28.1 (lactam CH2), 2 1.6 (CH ) .
HRMS (+ESI) C1 H1 N 0 2 + H+: calcd 247.1441 found 247.1453.
Reference Example 5 : 3-(4'-Chlorobenzoylamino)tetrahydropyridin-2-one:
3-aminotetrahydropyridin-2-one hydrochloride (10 mmol), 2C0 3 (30 mmol) and 4-
chlorobenzoyl chloride (10 mmol) were reacted according to the above procedure to
give the product (0.87 g, 39%):
vmax/cm 3295, 3202 ( -H, amide), 1668, 1648, 1629 (secondary CONH, lactam),
1594, 1486 (aromatic ring), 859, 845, 808, (para-disubstituted benzene ring), 750, 656
(C-Cl).
Ή NMR: dH (400MHz, CDC13) 7.74 (2H, br d, J 8.5, ortho-H), 7.38 (2H, br d, J 8.5,
et - ), 7.14 (1H, br d, J 3.0, C H4C1-C0N H), 5.83 ( 1H, br s, CONH-CH2), 4.40
(1H, dt, J 11.0, 5.5, CH-CO), 3.44-3.33 (2H, m, CH2NH), 2.73 (1H, dq, J 13.0, 4.5,
NHCH-C H ), 2.05-1.93 (2H, m, lactam CH ), 1.66-1.56 (1H, m, lactam CH2).
1 C NMR: C (100MHz, CDC1 ) 180.2 (lactam C=0), 171 .8 (aryl C=0), 138.1 (ipso-
C), 132.7 (C-Cl), 129.1 (aromatic-CH), 128.5 (aromatic-CH), 51.4 (CH-CO), 42.0
(CH2-NH), 27.2 (lactam CH ), 21.2 (lactam CH ) .
HRMS (+ESI) Ci2H,3ClN20 2 + Na+: calcd 275.0558; found 275.0559.
Reference Example 6: 3-(4'-Chlorobenzoylamino)azepan-2-one:
25
P66854.WO01 .Spec as filed 8.06.1
3-aminoazepan-2-one hydrochloride (10 mmol), 2C0 3 (30 mmol) and 4-
chlorobenzoyl chloride (10 mmol) were reacted according to the above procedure to
give the product ( 1.79 g, 75%):
vmax/cm 3243, 3200 (N-H, amide), 1662, 1643 (secondary CONH, lactam), 1595,
1484 (aromatic ring), 856, 841 , 819 (para-disubstituted benzene ring), 776, 732 (CCl).
NMR: dH (400MHz, CDC13) 7.76 (2H, br d, J 8.5, ortho-H), 7.59 (IH, br d, J 4.0,
C H4C1-C0NH), 7.39 (2H, br d, J 8.5, meta-H), 6.00 ( 1H , br s, CONH-CH2), 4.67
(IH, ddd, J 11.0, 5.5, 1.5, CH-CO), 3.39-3.22 (2H, , CH2NH), 2.22 (IH, br d, J 14.0,
lactam CH2), 2.09-2.00 (IH, m, lactam CH2), 1.96-1 .82 (2H, m, lactam CH2), 1.60-
1.36 (2H, m, lactam CH2) .
1 C NMR: C (100MHz, CDC13) 175.9 (lactam C=0), 165.3 (aryl C=0), 137.9 (ipso-
C), 132.7 (C-Cl), 128.9 (aromatic-CH), 128.7 (aromatic-CH), 52.8 (CH-CO), 42.3
(CH2-NH), 31.7 (lactam CH2), 29.0 (lactam CH ), 28.1 (lactam CH2).
HRMS (+ESI) C13H,5C1N20 2+ H+: calcd 267.0895; found 267.0890.
Reference Example 7 : 3-(4'-Methoxybenzoylamino)tetrahydropyridin-2-
one:
3-aminotetrahydropyridin-2-one hydrochloride ( 0 mmol), 2C0 (30 mmol) and 4-
methoxybenzoyl chloride (10 mmol) were reacted according to the above procedure to
give the product (2.18 g, 98%):
vma /cm 3305, 3212 (N-H, amide), 2854 (0-CH 3), 1693, 1627 (secondary CONH,
lactam), 1605, 1576, 1505 (aromatic ring), 837 ( -disubstituted benzene ring).
NMR: dH (400MHz, CDC13) 7.76 (2H, br d, J 9.0, ortho-H), 7.19 (IH, br d, J 5.0,
C7H7O -CONH), 6.87 (2H, br d, J 9.0, meta-H), 6.31 (IH, br s, CONH-CH2), 4.39 (IH,
dt, J 11.5, 5.5, CH-CO), 3.81 (3H, s, OCH3), 3.39-3.28 (2H, m, CH2NH), 2.64 (IH, dq,
26
P66854.WO01 .Spec as filed 8.06.1 1
J 13.0, 4.5, NHCH-C H ), 2.01-1 .88 (2H, m, lactam CH2), 1.68-1.55 (IH, m, lactam
CH2) .
C NMR: c (100MHz, CDC1 ) 172.4 (lactam C=0), 167.3 (aryl C=0), 162.4 (COCH
3), 129.1 (aromatic-CH), 126.6 (ipso-C), 113.8 (aromatic-CH), 55.6 (OCH 3), 5 1. 1
(CH-CO), 4 1.9 (CH2-NH), 27.4 (lactam CH2), 21.3 (lactam CH2).
HRMS (+ESI) C13H1 N20 + Na+: calcd 271.1053; found 271.1057.
Reference Example 8: 3-(4'-Methoxybenzoylamino)azepan-2-one:
3-aminoazepan-2-one hydrochloride (10 mmol), 2C0 3 (30 mmol) and 4-
methoxybenzoyl chloride (10 mmol) were reacted according to the above procedure to
give the product (1.54 g, 65%):
max m 1 3270, 3205 (N-H, amide), 2839 (0-CH ), 1642 (secondary CONH, lactam),
1602, 1577, 1504 (aromatic ring), 854, 822 (para-disubstituted benzene ring).
NMR: dH (400MHz, CDC13) 7.79 (2H, br d, J 9.0, ortho-H), 7.52 (IH, br d, J 5.0,
C7H70-CON H), 6.91 (2H, br d, J 9.0, meta-H), 5.94 (IH, br s, CONH-CH2), 4.69 (IH,
ddd, J 11.0, 5.5, 1.5, CH-CO), 3.83 (3H, s, OCH3), 3.40-3.21 (2H, , CH2NH), 2.22
(IH, br d, J 12.5, lactam CH2), 2.08-1.97 (IH, m, lactam CH2), 1.95-1.82 (2H, m,
lactam CH2), 1.60-1.36 (2H, , lactam CH2) .
1 C NMR: C (100MHz, CDC13) 176.2 (lactam C=0), 165.9 (aryl C=0), 162.1 (COCH
3), 129.1 (aromatic-CH), 126.7 (ipso-C), 113.8 (aromatic-CH), 55.5 (OCH 3), 52.7
(CH-CO), 42.3 (CH2-NH), 31.9 (lactam CH ), 29.1 (lactam CH2), 28.1 (lactam CH2) .
HRMS (+ESI) C14Hi N20 3 + Na+: calcd 285.1210; found 285.1215.
Reference Example 9 : 3-(4'-Fluorobenzoylamino)tetrahydropyridin-2-one:
27
P668S4.WO01 Spec as filed 8.06.1
3-aminotetrahydropyridin-2-one hydrochloride (10 mmol), 2C0 (30 mmol) and 4-
fluorobenzoyl chloride (10 mmol) were reacted according to the above procedure
(except that CHC13 was used instead of CH2C 12) to give the product ( 1.41 g, 54%):
vmax/cm 3216, 3075 (N-H, amide), 1650, 1555 (secondary CONH, lactam), 1595,
1491 (aromatic ring), 809, 844 (para-disubstituted benzene ring), 1105, 1 58, 1226,
1328, 756 (C-F).
NMR: H (400MHz, CDC1 ) 7.76 (2H, br dd, J 9.0, 5.5, ortho-H), 7.21 (IH, br s,
C H4F-CON H), 7.02 (2H, br t, J 8.5, met - ), 6.08 ( 1H, br s, CONH-CH2), 4.35 (IH,
dt, J 11.5, 5.5, CH-CO), 3.40-3.26 (2H, m, CH2NH), 2.62 (IH, dq, J 13.0, 4.5, NHCHCH
2), 2.00-1.86 (2H, , lactam CH2), 1.66-1.52 (IH, m, lactam CH2).
1 C NMR: c (100MHz, CDC13) 172.2 (lactam C=0), 166.6 (aryl C=0), 164.9 (d, J
252.0, C-F), 130.4 (d, J 3.0, ipso-C), 129.7 (d, J 9.0, ortho-C), 115.6 (d, J 22.0, meta-
C), 5 1. 1 (CH-CO), 41.9 (CH 2-NH), 27.3 (lactam CH2), 2 1.3 (lactam CH ) .
HRMS (+ESI) Ci2Hi FN20 2 + H+: calcd 237.1034; found 237.1034.
Reference Example 10: 3-(4'-FIuorobenzoylamino)azepan-2-one:
3-aminoazepan-2-one hydrochloride (10 mmol), 2C0 (30 mmol) and 4-
fluorobenzoyl chloride (10 mmol) were reacted according to the above procedure
(except that CHC13 was used instead of CH2CI2) to give the product (0.86 g, 45%):
Vmax c 3205, 3056 (N-H, amide), 1544 (secondary CONH, lactam), 1599, 1501
(aromatic ring), 821, 858 ( ra-disubstituted benzene ring), 1164, 1222, 1291, 765,
696 (C-F).
NMR: dH (400MHz, CDC1 ) 7.84 (2H, br dd, J 9.0, 5.5, ortho-H), 7.57 (IH, br s,
C6H4F-CON H), 7.09 (2H, br t, J 8.5, meta-H), 5.94 (IH, br s, CONH-CH2), 4.67 ( 1H,
ddd , J 11.5, 5.5, 1.5, CH-CO), 3.40-3.22 (2H, m, CH NH), 2.26-2.19 (IH, m, lactam
28
P66854.WOO1.Spec as filed 8.06.1 1
CH ), 2.09-2.00 (IH, m, lactam CH2), 1.96-1.83 (2H, m, lactam CH2), 1.60-1.36 (2H,
m, lactam CH ).
, C NMR: C (100MHz, d6-DMSO) 174.3 (lactam C=0), 164.1 (aryl C=0), 163.9 (CF,
d J 247), 130.7 (ipso-C), 29.8 (ortho-C, d, J 7), 115.2 (meta-C, d, J 22), 52.0 (CHCO),
40.6 (CH2-NH), 30.6 (lactam CH2), 28.9 (lactam CH2), 27.7 (lactam CH2) .
HRMS (+ESI) C13H,5FN20 2 + H+: calcd 251.1 190; found 251 . 1 192.
Reference Example 11: 3-(Pyridin -3'-carbonylamino)tetrahydropyridin-2-
one:
Oxalyl chloride (20 mmol) was added to a solution of nicotinic acid (10 mmol) in
DCM (40 mL), along with one drop of catalytic DMF. The reaction mixture was
stirred for 16 h and then the solvent was removed under high vacuum. The resulting
crystals were dissolved in DCM (10 mL). In a separate flask, 3-
aminotetrahydropyridin-2-one hydrochloride (10 mmol) and C0 3 (30 mmol) were
added to water (30 mL) and stirred, giving a solution to which the acid chloride
solution was added. The reaction was worked-up as above to give the product (0. 10 g,
5%):
ma m 3257 (N-H, amide), 1642, 1541 (secondary CONH, lactam, NH), 1591, 1479
(aromatic pyridine ring).
NMR: dH (400MHz, CDC1 ) 9.03 (IH, d, J 2.0, 2'-aryl CH), 8.71 (IH, dd, J 5.0,
1.5, 6'-aryl CH), 8.12 (IH, dt, J 8.0, 2.0, 4'-aryl CH), 7.36 (IH, dd, J 8.0, 5.0, 5'-aryl
CH), 7.27 (IH, br d, J 2.0, C5H4N-CON H), 5.91 (IH, br s, CONH-CH2), 4.45 (IH, dt,
J 11.0, 5.5, CH-CO), 3.44-3.32 (2H, , CH NH), 2.72 (IH, dt, J 14.5, 4.5, NHCHC
2), 2.06-1.93 (2H, , lactam CH2), 1.70-1.54 (IH, m, lactam CH2).
1 C NMR: c (100MHz, CDC13) 171.8 (lactam C=0), 166.0 (aryl C=0), 152.5 (aryl NCH),
148.6 (aryl N-CH), 135.3 (ortho-C(-CH)), 133.4 (ipso-C), 123.6 (meta-C), 51.2
(CH-CO), 42.0 (CH2-NH), 27.3 (lactam CH2), 21.3 (lactam CH2) .
29
P66854. OO Spec as filed 8.06.1
HRMS (+ESI) C,,H 13N 0 2 + H+: calcd 220.1081 ; found 220.1085.
Reference Example 12: 3-(Pyridin-3'-carbonyIamino)azepan-2-one:
Oxalyl chloride ( 1.69 mL, 20 mmol) was added to a solution of nicotinic acid ( 1.23 g,
10 mmol) in DCM (40 mL), along with one drop of catalytic DMF. The reaction
mixture was stirred for 16 h and then the solvent was removed under high vacuum.
The resulting crystals were dissolved in DCM (10 mL). In a separate flask, 3-
aminoazepan-2-one hydrochloride (10 mmol) and 2C0 3 (30 mmol) were added to
water (30 mL) and stirred, giving a solution to which the acid chloride solution was
added. The reaction was worked-up as above to give the product (0.66 g, 42%):
vmax cm 1 3200 (N-H, amide), 1642, 1548 (secondary CONH, lactam), 1590, 1476
(aromatic pyridine ring).
Ή NMR: dH (400MHz, CDC1 ) 9.06 (1H, d, J 2.0, 2'-aryl CH), 8.72 (1H, dd, J 5.0,
1.5, 6'-aryl CH), 8.1 1 (1H, dt, J 8.0, 2.0, 4'-aryl CH), 7.72-7.62 (1H, m, C5H4NCONH),
7.37 (1H, dd, 8.0, 5.0, 5'-aryl CH), 5.99 (1H, br s, CONH-CH2), 4.70 (1H,
ddd, J 11.0, 5.5, 1.5, CH-CO), 3.40-3.23 (2H, , CH2NH), 2.24 (1H, br d, J 14.5,
lactam CH ), 2.1 1-2.02 (1H, m, lactam CH ), 1.97-1.83 (2H, , lactam CH2), 1.63-
1.38 (2H, , lactam CH ) .
1 C NMR: C (lOOMHz, CDC13) 175.7 (lactam C=0), 164.6 (aryl C=0), 152.4 (aryl NCH),
148.5 (aryl N-CH), 135.1 (ortho-C(-CH)), 130.0 (ipso-C), 123.5 (meta-C), 52.8
(CH-CO), 42.2 (CH2-NH), 3 1.6 (lactam CH2), 28.9 (lactam CH2), 28.1 (lactam CH2) .
HRMS (+ESI) C12H i5N 0 2 + H+: calcd 234.1237; found 234.1239.
Reference Example 13: 3-(3',5'-Dimethylbenzoylamino)tetrahydropyridin-2-
one:
30
P66854.WO01 .Spec as filed 8.06.
3-aminotetrahydropyridin-2-one hydrochloride (10 mmol), K2C0 3 (30 mmol) and 3,5-
dimethylbenzoyl chloride (10 mmol) were reacted according to the above procedure
(except that CHC13 was used instead of CH2C 1 ) to give the product (2.06 g, 94%):
Vmax cm 1 32 12 (N-H, amide), 1675, 627, 1534 (secondary CONH, lactam), 1598,
1491 (aromatic ring), 890, 865, 817 (weia-trisubstituted benzene ring).
NMR: dH (400MHz, CDC1 ) 7.39 (2H, s, ortho-H), 7.15 (IH, br d, J 4.5, C H9-
CONH), 7.09 ( 1H, s,para-U), 6.25 ( 1H, br s, CONH-CH2), 4.41 (IH, dt, J 1 .5, 5.5,
CH-CO), 3.43-3.29 (2H, m, CH2NH), 2.69 (IH, dq, J 13.0, 4.5, NHCH-C H?), 2.31
(6H, s, CH ), 2.03-1 .89 (2H, , lactam CH2), 1.66-1.53 (IH, m, lactam CH ) .
C NMR: C (100MHz, CDCI 3) 172.2 (lactam C=0), 168.0 (aryl C=0), 138.2 (ipso-
C), 134.3 et -C), 133.5 (aromatic CH), 125.1 (aromatic CH), 51.2 (CH-CO), 41.9
(CH2-NH), 27.2 (lactam CH2), 2 1.4 (CH3), 2 .2 (lactam CH2) .
HRMS (+ESI) C14H N20 2 + H+: calcd 247.1441; found 247.1455.
Reference Example 14: 3-(3',5'-Dimethylbenzoylamino)azepan-2-one:
3-aminoazepan-2-one hydrochloride (10 mmol), 2C0 3 (30 mmol) and 3,5-
dimethylbenzoyl chloride (10 mmol) were reacted according to the above procedure
(except that CHCI3 was used instead of CH2C 12) to give the product (2.26 g, 96%):
vmax/cm 3319 (N-H, amide), 1682, 1635 (secondary CONH, lactam), 1600, 1498
(aromatic ring), 888, 866, 828 (meto-trisubstituted benzene ring).
NMR: dH (400MHz, CDC1 ) 7.57 (IH, br d, J 5.0, C H -CONH), 7.42 (2H, s,
ortho-H), 7.10 ( 1H, s, para-U), 6.09 ( 1H, br s, CONH-CH2), 4.69 (IH, ddd, J 11.0,
6.0, 1.5, CH-CO), 3.40-3.20 (2H, m, CH2NH), 2.33 (6H, s, CH3), 2.21 (IH, br d, J
12.5, lactam CH2), 2.08-1.98 (IH, m, lactam CH2), 1.97-1 .82 (2H, m, lactam CH2),
1.59-1.35 (2H, m, lactam CH2) .
3 1
P66854.WO01 .Spec as filed 8.06.1 1
13C NMR: C (100MHz, CDC13) 176.0 (lactam C=0), 166.8 (aryl C=0), 138.3 (ipso-
C), 134.3 (rneta-C), 133.3 (aromatic CH), 124.9 (aromatic CH), 52.6 (CH-CO), 42.3
(CH2-NH), 31.8 (lactam CH2), 29.0 (lactam CH2), 28.1 (lactam CH2), 21.4 (CH ) .
HRMS (+ESI) C 5H20N2O + H+: calcd 261.1598; found 261.1602.
In the examples below, the general procedure for the synthesis of 3-acylamino-2-
oxopiperidines was: potassium carbonate (3 mmol) and (5)-3-amino-2-oxopiperidine
hydrochloride (1.5 mmol) were dissolved in water (5 ml) and the solution was cooled
to 0 °C, and a solution of substituted benzoyl chloride ( 1 mmol) in tetrahydrofuran (5
mL) was added. The mixture was stirred for 6 hours, and then the reaction was
extracted with dichloromethane or chloroform. The combined organic layers were
dried over sodium sulfate and reduced in vacuo to give a solid. This solid was
redissolved in a minimum amount of dichloromethane and crystallised by addition of
petroleum ether 40-60 °C. The solid product was isolated by filtration and dried over
potassium pentoxide.
Example 1: (S)-3-Fluoro-N-(2-oxopiperidin-3-yl)benzamide
0.147 g off-white coarse powder (41 %). p 164-166 °C. [a] D -5.50 (c 0.1, MeOH);
vmax/cm 1669, 1644 (C=0, amide), 1552 (N-H, amide), 1303 (C-F). Anal.
(C, 2H,3FN20 2) C, H, N: calcd C 61.01, H 5.55, N 11.86; found C 60.65, H 5.50, N
11.78. Ή-NMR dH. 7.52 (2H, tt, J 8 and 2 , ArH4 and ArH6), 7.38 ( 1H, td, J 8 and
5.5, ArH2), 7.21-7.14 (2H, m, NHCH and ArH3), 5.9 ( 1H, s, NHCH2), 4.41 (IH, dt, J
11.5 and 6 , CHNH), 3.42-3.28 (2H, , CH2NH), 2.72 (IH, dq, J 13 and 5 , CH2CH),
2.04-1.95 (2H, , G¾CH 2NH), 1.62 (IH, dq, J 16 and 5 , CH2CH). 1 C-NMR C
32
P66854.WOO 1.Spec as filed 8.06. 11
171.6 (CHCONH), 166.3 (CONHCH), 162.8 (d, J 247, ArC3), 136.4 (CCO), 130.17
(d, J 12, ArC5), 122.6 (ArC6), 118.6 (d, J 21, ArC2), 114.6 (d, J 21, ArC4), 5 1.2
(CHNH), 41.8 (CH2NH), 27.0 (CH2CHNH), 21.0 (CH2CH2NH). F-NMR F - 1 11.9.
HRMS (+ESI) Ci2Hi FN20 2Na: calcd 259.0853; found 259.0859.
Example 2: (S)-2-FIuoro-N-(2-oxopiperidin-3-yl)benzamide
0.222 g white fluffy powder (63 %). mp 166-168 °C. [a] D -10.00 (c 0.1, MeOH).
Vmax cm 1 1647, 1613 (C=0, amide), 1512 (N-H, amide), 1277 (C-F). Anal.
(C12Hl3FN 0 2) C, H, N: calcd C 61.01, H 5.55, N 11.86; found C 60.78, H 5.54, N
.69. -NMR dH. 8.05 (IH, td, J 8 and 2 , ArH6), 7.62 ( 1H, dd, J 6 and 4, NHCH),
7.45 (IH, dddd, J 8,7, 5 and 2, ArH4), 7.21 (IH, td, J 7.5 and 1, ArH5), 7.13 ( 1H, ddd,
J 12, 9 and 1, ArH3), 6.01 ( 1H, s, NHCH ), 4.53 (IH, dt, J 11 and 6, CHNH), 3.45
(2H, td, J 6 and 3, CH2NH), 2.72 (IH, dq, J 13 and 6, CH2CH), 2.02-1.95 (2H, ,
CH2CH2NH), 1.72-1.59 (IH, m, CH2CH). 1 C-NMR C 171.3 (CHCONH), 163.4
(CONHCH), 160.9 (d, J 248.5, ArC2), 130.17 (d, J 12, ArC5), 133.4 (d, J 9, ArC4),
131.9 (ArC6), 120.9 (d, J 9, CCO), 116.1 (d, J 21, ArC3), 51.3 (CHNH), 41.9
(C¾NH), 27.2 (CH2CHNH), 21.1 (CH CH2NH). 1 F-NMR F - 112.4. HRMS (+ESI)
C 2Hi3FN20 2Na: calcd 259.0853; found 259.0858.
Example 3: (S )-4-Fluoro-N-(2-oxopiperidin-3-yl)benzamide
0.145 g cream coloured fine crystals (41 %)mp 133-135 °C; [a] D -7.90 (c 0.1,
MeOH); v„/cm ' 1650, 1636 (C=0, amide), 1557 (N-H, amide), 1327 (C-F) Anal.
33
P66854.WO01 .Spec as filed 8.06.1
(Ci 2H1 FN20 2) C, H, N: calcd C 61.01, H 5.55, N 11.86; found C 60.71, H 5.38, N
11.44 (1/3 H20). -NMR dH 7.98 (2H, q, J 5, ArH3 and ArH5), 7.41 ( 1H, d, J 6,
NHCH), 7.20 (2H, tt, J 9,2, ArH2 and ArH6), 6.31 ( 1H, s, NHCH2), 4.52 (IH, dt, J
11 and 6, CHNH), 3.52 (2H, td, J 6 and 2, CH2NH), 2.84 (IH, dq, J 13 and 4,
CH2CH), 2.19-2.10 (2H, , CH2CH2NH), 1.81 (IH, dq, J 12 and 8, CH2CH). 1 CNMR
C 171.7 (CHCONH), 166.6 (CONHCH), 164.9 (d, J 251.5, ArC4), 130.3 (d, J
4, CCO), 129.5 (d, J 9, ArC2/6), 115.5 (ArC3/5), 51.2 (CHNH), 4 1.8 (CH2NH), 27.1
(CH2CHNH), 21.0 (CH2CH2NH). HRMS (+ESI) C,2H13FN 0 2Na: calcd 259.0853;
found 259.0852.
Example 4: (S)-N-(2-Oxopiperidin-3-yl)-4-(trifluoromethyl)benzamide
0.376 g white fine powder (87 %). mp 212-214 °C; [a] -6.35 (c 0.1, MeOH);
ma cm 1 1650, 1636 (C=0, amide), 1557 (N-H, amide), 1327 (C-F). Anal.
(C, 3H,3F3N20 2) C, H, N: calcd C 54.55, H 4.58, N 9.79; found C 53.99, H 4.68, N
9.59. -NMR dH -NMR d H. 7.51 (2H, d, J 8, ArH2 and ArH6), 7.22 (2H, d, J 8.5,
ArH3 and ArH5), 7.15 ( 1H , s, NHCH), 5.70 (IH, s, NHCH2), 4.00 (IH, dt, J 11.5 and
6, CHNH), 2.98 (2H, td, J 6 and 2, CH2NH), 2.25 (IH, dq, J 13 and 4, CH2CH), .62-
1.53 (2H, m, CH2CH2NH), 1.28 (IH, dq, J 12 and 8, CH2CH). , C-NMR C 171.8
(CHCONH), 166.3 (CONHCH), 137.3 (ArCl), 133.3 (q, J 31, ArC4), 127.6 (ArC2/6),
125.5.9 (q, J 4, ArC3/5), 123.7 (q, J 271, CF ), 51.2 (CHNH), 4 1.9 (CH2NH), 27.0
(CH CHNH), 21.1 (CH2CH2NH). 1 F-NMR d -62.9. HRMS (+ESI) C,3Hi3F3N20 2Na:
calcd 309.0821; found 309.0822.
Example 5 : (5)-N-(2-Oxopiperidin-3-yl)-3-(trifluoromethyl)benzamide
34
P66854.WO0l .Spec as filed 8.06.1
0.251 g cream fine powder (59 %). mp 148-150 °C; [a] 4
D -10.20 (c 0.1, MeOH);
vmax/cm ' 1671, 1648 (C=0, amide), 1554 (N-H, amide), 1327 (C-F). Anal.
(Ci3H,3F3 20 2) C, H, N: calcd C 54.55, H 4.58, N 9.79; found C 53.90, H 4.56, N
9.60. -NMR dH 7.80 (IH, s, ArH2), 7.73 ( 1H , d, J 7, ArH4), 7.60 ( 1H, d, J 6,
NHCH), 7.43 (IH, d, J 7, ArH6), 7.23 ( 1H , d, J 8, ArH5), 6.52 ( 1H, s, NHCH2), 4.23
(IH, dt, J 11 and 6, CHNH), 3.17-3.06 (2H, , CH2NH), 2.30 (IH, dq, J 13 and 6,
CH2CH), 1.74-1.65 (2H, m, CH2CH2NH), 1.62-1.42 (IH, m, GH2CH). 1 C-NMR C
171.6 (CHCONH), 166.1 (CONHCH), 134.9 (ArCl), 131.1 (q, J 34, ArC3), 130.3
(ArC5), 129.1 (ArC6), 128.2 (q, J 4, ArC2), 124.3 (q, J 4, ArC4), 123.7 (q, J 271,
CF3), 51.2 (CHNH), 41.8 (CH2NH), 27.1 (CH2CHNH), 21.1 (CH2CH NH). 19F-NMR
F -62.7. HRMS (+ESI) Ci3H13F N20 2Na: calcd 309.0821; found 309.0820.
Example 6 : (5)-N-(2-Oxopiperidin-3-yI)-2-(trifluoromethyl)benzamide
0.262 g white fluffy powder (61 %). mp 155-156 °C; [a] 24
D -18.20 (c 0.1, MeOH);
vmax/cm 1674, 1654 (C=0, amide), 1543 (N-H, amide), 1312 (C-F). Anal.
(C, 3H, F3N 0 ) C, H, N: calcd C 54.55, H 4.58, N 9.79; found C 54.25, H 4.51, N
9.70. 1H-NMR dH 7.64 (IH, d, J 7, ArH6), 7.54 ( 1H, d, J 6, ArH3), 7.22-7.15 (2H, m,
ArH4 and ArH5), 6.78 ( 1H , d, J 5, NHCH), 6.08 (IH, NHCH2), 4.42 (IH, dt, J 11 and
6, CHNH), 3.35 (2H, td, J 6 and 2, CH2NH), 2.73 (IH, dq, J 13 and 6, CH CH), 1.99-
1.91 (2H, , CH2CH2NH), 1.59 (IH, dq, J 12 and 8, CH2CH). 1 C-NMR C 171.0
(CHCONH), 167.8 (CONHCH), 135.5 (ArCl), 131 .9 (ArC4), 129.9 (ArC5), 128.6
(ArC6), 127.4 (q, J 31, ArC2), 126.4 (q, J 4, ArC3), 123.6 (q, J 270, CF ), 51.3
35
P66854.WO0I .Spec as filed 8.06. 1
(CHNH), 41.8 (CH2NH), 26.5 (CH2CHNH), 20.9 ( H2CH2NH). 1 F-NMR F -58.7.
HRMS (+ESI) C 3H F N20 2Na: calcd 309.0821; found 309.0818.
Example 7: (5)-2,3-difluoro-N-(2-oxopiperidin-3-yl)benzamide
0.234 g white needle crystals (61 %). mp 160-162 °C; [a] 2
D -5.65 (c 0.1, MeOH);
Vmax c 1 1686, 1648 (C=0, amide), 1536 (N-H, amide), 1320 (C-F). Anal.
(C, 2Hi2F2N20 2) C, H, N: calcd C 56.99, H 4.76, N 11.02; found C 56.26, H 4.69, N
10.84. -NMR H. 7.76 (1H, ddt, J 8, 6 and 2, ArH6), 7.51 ( 1H, q, J 3, NHCH), 7.28
(1H, dddd, J 9.5, 8, 7.5, 2, ArH4), 7.15 ( 1H, tdd, J 8, 5, 1.5, ArH5), 6.18 ( 1H , s,
NHC¾), 4.47 ( 1H, dt, J 12 and 4, CHNH), 3.38 (2H, td, J 6 and 2, CH2NH), 2.69 (1H,
dq, J 12.5 and 3.5, CH2CH), 2.02-1.93 (2H, m, CH2CH2NH), 1.73-1 .59 (1H, m,
CH2CH). 1 C-NMR C 171.2 (CHCONH), 162.5 (CCONH), 132.8 (dd, J 250 and 15,
ArC3), 149.1 (dd, J 251 and 14, ArC2), 126.2 (t, J 3, ArC5), 124.4 (t, J 4, ArC6),
123.3 (d, J 9, CCONH), 120.3 (d, J 17, ArC4), 1.4 (CHNH), 41.8 (CH2NH), 27.1
(CH2CHNH), 21.1 (CH CH2NH). HRMS (+ESI) Ci2H 2F N20 2Na: calcd 277.0759;
found 277.0769.
Example 8 : (S )-2,4-difIuoro-N-(2-oxopiperidin-3-yI)benzamide
0.236 g off-white crystals (62 %). mp 140-141 °C; [a] 4
D -2.00 (c 0.1, MeOH);
vmax/cm 1682, 1638 (C=0, amide), 1526 (N-H, amide), 1289 (C-F). Anal.
(C12H12F2N20 2.l/6 H20 ) C, H, N: calcd C 56.03, H 4.83, N 10.89; found C 56.40, H
4.69, N 10.93. -NMR H. 8.07 (1H, td, J 9 and 7, ArH6), 7.54 ( 1H, q, J 5, NHCH),
36
P66854.WO0 .Spec as filed 8.06. 1
6.95 (IH, tdd, J 8, 2.5, 1, ArH5), 6.84 (IH, dq, J 8.5, 2, ArH3), 6.43 ( 1H, s, NHCH2),
4.45 (IH, dt, J 1 and 6, CHNH), 3.36 (2H, td, J 6.5 and 2.5, CH2NH), 2.66 (IH, dq, J
12.5 and 5.5, CH2CH), 2.00-1.91 (2H, , CH2CH2NH), 1.73-1 .59 (IH, , CH2CH).
1 C-NMR c 171.4 (CHCONH), 162.5 (CCONH), 164.9 (dd, J 255 and 12, ArC2),
161.2 (dd, J 253 and 12, ArC4), 133.6 (dd, J 10 and 4, ArC6), 17.4 (dd, J 12 and 4,
CCONH), 112.2 (dd, J 2 1 and 3, ArC5), 104.3 (t, J 27, ArC3), 51.3 (CHNH), 41.8
(CH NH), 27.2 (CH2CHNH), 21.1 (CH CH2NH). HRMS (+ESI) C,2H,2F2 20 2Na:
calcd 277.0759; found 277.0761.
Example 9 : (S)-2,5-difluoro-N-(2-oxopiperidin-3-yl)benzamide
0.252 g off-white crystals (66 %). p 140-142 °C; [a] D +0.85 (c 0.1, MeOH);
v cm 1681, 1635 (C=0, amide), 1519 (N-H, amide), 1328 (C-F). Anal.
(C,2H12F2N20 2.1/6 H20 ) C, H, N: calcd C 56.03, H 4.83, N 10.89; found C 55.47, H
4.72, N 10.67. -NMR dH. 8.07 (IH, dt, J 11 and 8.5, ArH6), 7.54 ( 1H, dd, J 11, 5,
NHCH), 6.68-6.54 (2H, , ArH3 and ArH4), 6.43 ( 1H, s, NHCH ), 4.45 (IH, dt, J 11
and 6, CHNH), 3.36 (2H, td, J 6.5 and 2.5, CH2NH), 2.66 (IH, dq, J 12.5 and 5.5,
CH2CH), 1.99-1 .90 (2H, , CH2CH2NH), .64 ( 1H, dq, J 11 and 8 CH2CH). 1 C-NMR
c 171.3 (CHCONH), 162.2 (CCONH), 158.6 (d, J 258, ArC5), 156.7 (d, J 258,
ArC2), 133.6 (dd, J 10 and 4, ArC6), 122.4 (dd, J 8 and 6, CCONH), 120.0 (dd, J 24
and 8, ArC3), 118.0 (dd, J 26 and 4, ArC4), 117.5 (dd, J 28 and 7, ArC6), 51.4
(CHNH), 41.8 (CH2NH), 27.1 (CH2CHNH), 21.1 (CH2CH NH). HRMS (+ESI)
C12H12F2N20 2Na: calcd 277.0759; found 277.0766.
Example 10: (S)-2,6-difluoro-N-(2-oxopiperidin-3-yl)benzamide
37
P66854.WOO Spec as filed 8.06.1 1
0.209 g white fine powder (55 %). mp 190-194 °C; [a] 2 -7.20 (c 0.1, MeOH);
vmax/cm 1672, 1638 (C=0, amide), 1492 (N-H, amide), 1329 (C-F). Anal.
(Ci2H| F2N 02) C, H, N: calcd C 56.99, H 4.76, N 11.02; found C 55.93, H 4.68, N
10.83 (1/6 H20). -NM dH. 7.30 (IH, tt, J 9 and 6, ArH4), 7.06 (IH, d, J 3.5,
NHCH), 6.89 (2H, t, J 3.5, ArH3 and ArH5), 6.23 ( 1H, s, NHCH2), 4.43 (IH, dt, J 11
and 5, CHNH), 3.37-3.32 (2H, , CH NH), 2.75 (IH, dq, J 14.5 and 4.5, CH2CH),
.99-1.90 (2H, , CH2CH2NH), 1.68-1.54 (IH, , CH2CH). 1 C-NMR C 171.1
(CHCONH), 160.5 (CCONH), 160.1 (dd, J 250 and 7.5, ArC2 and ArC6), 131.7 (t, J
I I , ArC4), 114.1 (t, J 20, CCONH), 111.9 (dd, J 20 and 5, ArC3 and ArC5), 51.3
(CHNH), 41.7 (CH NH), 26.8 (CH2CHNH), 20.9 (CH2CH2NH). 1 F-NMR
- 1 12.5. HRMS (+ESI) C 2H,2F2N20 2Na: calcd 277.0759; found 277.0760.
Example 11: (S)-3,4-difluoro-N-(2-oxopiperidin-3-yl)benzamide
0.226 g off-white fine powder (59%). mp 202-204 °C; [a] 2
D -7.50 (c 0. , MeOH);
vma /cm 1691, 1652 (C=0, amide), 1487 (N-H, amide), 1285 (C-F). Anal.
(Ci 2Hi2F2N20 2) C, H, N: calcd C 56.99, H 4.76, N 1.02; found C 56.06, H 4.68, N
10.88 (1/6 H20). -NMR d H. 7.66 (IH, qd, J 7 and 2, ArH2), 7.56-7.50 ( 1H, m,
ArH5), 7.24-7.14 (2H, m, ArH6 and NHCH), 5.90 (IH, s, NHCH2), 4.38 (IH, dt, J
11.5 and 5, CHNH), 3.39 (2H, td, J 7 and 3, CH2NH), 2.69 (IH, dq, J 14 and 5,
CH2CH), 2.03-1 .94 (2H, , CH2CH2NH), 1.68-1 .54 (IH, , CH2CH). 1 C-NMR C
171 .5 (CHCONH), 165.4 (CCONH), 152.5 (dd, J 253 and 12, ArC3), 150.2 (dd, J 249
and 12, ArC4), 131.2 (t, J 4.5, CCONH), 123.5 (q, J 3.5, ArC6), 117.4 (d, J 18, ArC5),
38
P66854.WOO 1.Spec as filed 8.06.1 1
116.9 (d, J 18, ArC2), 51.2 (CHNH), 41.8 (CH2NH), 26.9 (CH2CHNH), 21.1
(CH2CH2NH). HRMS (+ESI) C12H1 F2N20 Na: calcd 277.0759; found 277.0751.
Example 12: (S )-3,5-difluoro-N-(2-oxopiperidin-3-yl)benzamide
0.234 g white fine powder (61 %). p 198-199 °C; [a] 4
D -9.50 (c 0.1, MeOH);
ma c n 1 1674, 1641 (C=0, amide), 1565 (N-H, amide), 1333 (C-F). Anal.
(C 2H F N20 2) C, H, N: calcd C 56.99, H 4.76, N 11.02; found C 56.20, H 4.69, N
10.93. -NMR dH. 7.31 (2H, dd, J 8 and 2.5, ArH2 and ArH6), 7.26 ( 1H , d, J 5,
NHCH), 6.9 (IH, tt, J 9 and 2, ArH4), 6.01 (IH, s, NHCH2), 4.39 (IH, dt, J 12 and
5.5, CHNH), 3.41-3.36 (2H, m, CH2NH), 2.68 (IH, dq, J 12 and 5, CH2CH), 2.03-1.94
(2H, , CH2CH2NH), 1.69-1.55 (IH, m, CH CH). I C-NMR C 171.3 (CHCONH),
165.2 (CCONH), 162.9 (dd, J 251.5 and 12, ArC3 and ArC5), 137.5 (t, J 9, CCONH),
110.4 (dd, J 20 and 7, ArC2 and ArC6), 107.0 (t, J 20, ArC4), 51.3 (CHNH), 41.8
(CH2NH), 26.9 (CH2CHNH), 20.9 ( H2CH2NH). 1 F-NMR F -108.2. HRMS (+ESI)
C, H, F2N20 2Na: calcd 277.0759; found 277.0750.
Example 13: (S)-3-(3'-Butylbenzoylamino)-azepan-2-one
(5)-3-Amino-azepan-2-one hydrochloride (0.72 g, 4.39 mmoles) was dissolved in H20
(20 mL) and cooled to 0°C. 3-Butylbenzoyl chloride in dichloromethane was added
and triethylamine (1.3 mL, 9 mmoles) and the reaction was stirred over night. H20 (20
mL) was added and the reaction was extracted with dichloromethane (3 20 mL), the
39
P66854.WO0l .Spec as filed 8.06. 11
organic layer was washed with a pH 2 buffer (3 15 mL), dried over Na S04 and
reduced in vacuo. The product was purified by silica column chromatography
(petroleum ethenethyl acetate 75:25 to 0:100) to give the product as a white solid H
(400 MHz, CDC1 ) 7.71-7.62 (m, 3H, Ar & NHCH), 7.35-7.29 (m, 2H, Ar), 7.06 (br.t,
IH, J 6, NHCH2), 4.72 (dd, IH, J 11, 6, NHCH), 3.37-3.32 (m, 2H, NHCH ), 2.65 (t,
2H, J 8, CH3CH CH2CH ) , 2.22 (br.d, 1H, J 13.5, NHCHC H ), 2.03 (br.d, 1H, J 14,
NHCHCH 2CH ) , 1.95-1.81 (m, 2H, NHCHCH 2CH CH ), 1.61 (quintet, 2H, J 7,
CH CH2CH ) , 1.57-1 .48 ( , 1H. NHCHCH^), 1.46-1 .25 (m, 3H, NHCHCHzC^C H
and CH3CH ) and 0.92 (t, 3H, J 7.5, H8); C (100 MHz, CDC13) 176.0 (C=0), 166.6
(C=0), 143.4 (Ar quat, C- Bu), 134.2 (Ar quat, C-C=0), 128.4 (CH, Ar), 127.2 (CH,
Ar), 126.8 (CH, Ar), 124.0 (CH, Ar), 52.6 (CH-NH), 42.1 (CH2-NH), 35.6 (CH2), 33.5
(CH2), 31.6 (CH2), 28.9 (CH ), 28.0 (CH ), 22.3 (CH2) and 13.9 (CH3).
Example 14: (S)-3-(4'-Ethylbenzoylamino)-azepan-2-one
(5)-3-amino-azepan-2-one hydrochloride (1.65 g, 10 mmoles) was dissolved in H20
(20 mL) and cooled to 0°C. 4-Ethylbenzoyl chloride in dichloromethane was added
and triethylamine (4.2 mL, 30 mmoles) and the reaction was stirred over night. H20
(20 mL) was added and the reaction was extracted with dichloromethane (3 c 20 mL),
the organic layer was washed with a pH 2 buffer (3 15 mL), dried over Na2S0 4 and
reduced in vacuo. The product was purified by recrystallisation from chloroform and
cold petroleum ether to give the product as a white solid 0.94 g (36 %); mp 218-219 0
C; dH (400 MHz, CDC13) 7.66 (d, 2H, J 8, CH-C-Et), 7.62 (d, 1H, J 5.5, NH-CH), 7.24
(d, 2H, J 8, CH-C-CO), 6.55 (br.t, 1H, J 6, NH-Cl), 4.70 (dd, 1H, J 11, 5.5, CH-C4),
3.37-3.32 (m, 2H, HI), 2.67 (q, 2H, J 7.5, H5), 2.21 (br.d, IH, J 13, H4 equatorial),
2.02 (dt, IH, J 14, 4, H3 equatorial), 1,95-1.82 (m, 2H, H2 equatorial & H3 axial),
1.53 (br.q, 1H, J 12.5, H4 axial), 1.40 (br.q, 1H, J 13, H2 axial) and 1.22 (t, 3H, J 7.5,
H6); C (100 MHz, CDC13) 175.9 (C=0), 166.2 (C=0), 148.2 (C-Et), 131 .6 (C-C=0),
128.0, 127.2 (CH phenyl), 52.6 (CH-NH), 42.2 (CI), 28.9 (C5), 28.8, 28.0 (C2, C3)
40
P66854.WO0l .Spec as filed 8.06.1 1
and 5.4 (C6); /cm 1: 3200 (NH indole), 2956 (C-H), 1642 (amide C=0) and 1543
(aromatic); ESI m/z 100 %, 542.9 (M2Na+), 70%, 283.1 (M a+) and 10 %, 261 .2
(MH+); HR ESI m/z (Ci 5H2oN20 2Na requires 283.1417) found 283.1414; [a] (c =
0.49, CHC1 ) +70.48.
Example 15: (S)-3-(4'-Ethylbenzoylamino)-tetrahydropyridin-2-one
(5)-3-amino-tetrahydropyridin-2-one (20 mmoles) was dissolved in H20 (100 mL) and
cooled to 0°C. 4-Ethylbenzoyl chloride (16 mmoles) in dichloromethane was added
and triethylamine (6.7 mL, 48 mmoles) and the reaction was stirred over night. The
reaction was extracted with dichloromethane (3 x 30 mL), the organic layer was
washed with a pH 2 buffer (3 20 mL), dried over Na2S0 4 and reduced in vacuo. The
product was purified by silica column chromatography (petroleum ether:ethyl acetate
50:50:0 to 0:80:20) to give the product as a white solid 1.46 g (37 %); mp 112-1 13 0 C;
dH (400 MHz, CDCI3) 7.71 (d, 2H, J 8.5, CH-C-Et), 7.55 (d, 1H, J 5.5, NH-CH), 7.19
(d, 2H, J 8.5, CH-C-CO), 6.69 (br.s, 1H, NH-C1), 4.39 (dt, 1H, J 11, 5.5, CH-C3),
3.35-3.28 (m, 2H, HI), 2.67 (q, 2H, J 7.5, H4), 2.63-2.56 ( , 1H, H3 equatorial), 1.94-
1.87 (m, 2H, H2), 1.68 (tt, 1H, J 12.5, 8, H3 axial), and 1.20 (t, 3H, J 7.5, H5); C (100
MHz, CDCI3) 172.2 (C=0), 167.5 ( =0 ), 148.2 (C-Et), 131.5 (C-C=0), 127.9, 127.2
(CH phenyl), 50.9 (CH-NH), 41.7 (CI), 28.8 (C4), 27.2 (C3), 21.1 (C2) and 15.3 (C5);
Umax/cm 1 : 3334, 3245 (NH), 2932 (C-H), 1656, 1634 (C=0) and 1528 (aromatic); ESI
m/z 100 %, 514.9 (M2Na+) and 35%, 269.1 (MNa+); HR ESI m/z (C14Hi8N 0 2Na
requires 269.1260) found 269.1261; [a] D(c = 0.491, CHC13) +103.95.
Example 16: (S)-3-(4'-Butylbenzoylamino)-azepan-2-one
4 1
P66854. OO Spec as filed 8.06.1
(S)-3-amino-azepan-2-one hydrochloride (1.65 g, 10 mmoles) was dissolved in H20
(20 mL) and cooled to 0°C. 4-Butylbenzoyl chloride (8 mmoles) in dichloromethane
was added and triethylamine (4.2 mL, 30 mmoles) and the reaction was stirred over
night. H20 (20 mL) was added and the reaction was extracted with dichloromethane (3
20 mL), the organic layer was washed with a pH 2 buffer (3 5 mL), dried over
Na2S0 4 and reduced in vacuo. The product was purified by silica column
chromatography (petroleum ethenethyl acetate 75:25 to 0:100) to give the product as a
white solid 0.42 g (18 %); mp 183-184 0 C; d (400 MHz, CDC13) 7.73 (d, 2H, J 8,
Ar), 7.63 (d, 1H, J 5.5, NHCH), 7.21 (d, 2H, J 8, Ar), 6.80 (br.t, 1H, J 6, NHCH2),
4.69 (dd, 1H, J 10.5, 5.5, NHCH), 3.35-3.20 ( , 2H, CH^NH), 2.61 (t, 2H, J 7.5, H5),
2.19 (br.d, 1H, J 13.5, H4 equatorial), 2.00 (br.d, 1H, J 12.5, H3 equatorial), 1.93-1.80
(m, 2H, H2 equatorial & H3 axial), 1.70-1.46 (m, 3H, H6 and H4 axial), 1.38 (br.q,
1H, J 13, H2 axial), 1.3 1 (sextet, H2, J 7, H7) and 0.89 (t, 3H, J 7.5, H8); C (100
MHz, CDC13) 176.0 (C=0), 166.3 (C=0), 146.7 ( -nBu), 131.6 (C-C=0), 128.5, 127.1
(CH phenyl), 52.5 (CH-NH), 42.2 (CI), 35.5 (C5), 33.4 (C4), 31.6 (C6), 28.9, 28.0
(C2, C3), 22.3 (C7) and 13.9 (C6); /cm 1 : 3359, 3207 (NH), 2951 (C-H), 1671,
1650 (CO) and 1543 (aromatic); ESI m/z 100 %, 3 11.2 (M a+) and 22 %, 289.2
(MH+); HR ESI m/z (C, 7H24N20 2Na requires 3 1.1730) found 3 11.1732; [a] 5
D (c =
0.515, CHCI3) +64.88.
Example 17: (S)-3-(4'-Butylbenzoylamino)-tetrahydropyridin-2-one
42
P66854.WO0 Spec as filed 8.06.1 1
(5)-3-amino-tetrahydropyridin-2-one (20 mmoles) was dissolved in H 0 (20 mL) and
cooled to 0°C. The 4-butylbenzoyl chloride in dichloromethane was added and
triethylamine (4.2 mL, 30 mmoles) and the reaction was stirred over night. H20 (20
mL) was added and the reaction was extracted with dichloromethane (3 c 20 mL), the
organic layer was washed with a pH 2 buffer (3 15 mL), dried over Na S0 4 and
reduced in vacuo. The product was purified by silica column chromatography
(petroleum ethenethyl acetate: methanol 50:50:0 to 0:80:20) to give the product as a
white solid 0.45 g (16 %); dH (400 MHz, CDC1 ) 7.70 (d, 2H, J 8, CH-C- Bu), 7.36 (d,
1H, J 6, NH- mp 117-1 18 C; CH), 7.17 (d, 2H, J 8, CH-C-CO), 6.67 (br.s, 1H, NHCI),
4.69 (dt, 1H, J 12, 6, CH-C4), 3.36-3.29 (m, 2H, HI), 2.60 (t, 3H, J 7.5, H4 &
H3), 1.93-1.86 (m, 2H, H2), 1.67-1.54 (m, 1H, H3), 1.55 (quintet, 2H, J 7.5, H5), 1.30
(sextet, 2H, J 7, H7.5, H6) and 0.89 (t, 3H, J 7.5, H7); C (100 MHz, CDCI3) 172.2
(C=0 lactam), 167.5 (C=0 amide), 146.9 (C- Bu), 131.5 (C-C=0), 128.5 (CH phenyl),
127.2 (CH phenyl), 50.8 (CH-NH), 41.7 (CI), 35.5 (C4), 33.3 (C5), 27.2 (C3), 22.3
(C6), 21.1 (C2) and 13.9 (C7); ESI m/z 100 %, 297.2 (MNa +) and 26 %, 275.2 (MH+);
HR ESI m/z (C16H22N20 2Na requires 297.1573) found 297.1573; [a] (c = 0.523,
Example 18: (R)-3-(4'-Butylbenzoylamino)-tetrahydropyridin-2-one
(^)-3-amino-tetrahydropyridin-2-one (10 mmoles) was dissolved in H20 (30 mL) and
cooled to 0°C. 4-Butylbenzoyl chloride (8.5 mmoles) in dichloromethane was added
and triethylamine (4.2 mL, 30 mmoles) and the reaction was stirred over night. The
reaction was extracted with dichloromethane (3 * 15 mL), the organic layer was
washed with a pH 2 buffer (3 15 mL), dried over Na S0 4 and reduced in vacuo. The
product was purified by silica column chromatography (petroleum ethenethyl
acetate:methanol 50:50:0 to 0:80:20) to give the product as a white solid 1.10 g (40
%); mp 116-1 17 0 C; dH (400 MHz, CDCI3) 7.72 (d, 2H, J 8, CH-C- Bu), 7.25 (d, 1H, J
43
P668S4. OO Spec as filed 8.06. 11
5.5, NH-CH), 7.20 (d, 2H, J 8, CH-C-CO), 6.41 (br.s, 1H, NH-Cl), 4.41 (dt, 1H, J
11.5, 5.5, CH-C4), 3.37-3.32 (m, 2H, HI), 2.66 (ddt, IH, J 13, 6, 4.5, H3 equatorial),
2.62 (t, 3H, J 8, H4), 1.98-1.90 (m, 2H, H2), 1.67-1.53 (m, 3H, H3 axial & H5), 1.32
(quintet, 2H, J 7.5, H6) and 0.90 (t, 3H, J 7, H7); d (100 MHz, CDC13) 172.1 (C=0
lactam), 167.6 (C=0 amide), 147.0 (C-nBu), 131.5 (C-C=0), 128.5 (CH phenyl), 127.2
(CH phenyl), 51.0 (CH-NH), 4 1.7 (CI), 35.5 (C4), 33.3 (C5), 27.2 (C3), 22.3 (C6),
21.1 (C2) and 13.9 (C7); ^/cm : 3319, 3245 (NH), 2949 (C-H), 1651, 1634 (C=0)
and 1521 (aromatic); ESI m/z 100 %, 297.1 (MNa+); HR ESI m/z (Ci 6H2 N20 2Na
requires 297.1573) found 297.1574; [ ] D (c = 0.493, CHC1 ) -89.45.
Example 19: (S)-3-(4'-½ri-Butylbenzoylamino)-azepan-2-one
3 2
(5)-3-amino-azepan-2-one hydrochloride (2.04 g, 12.44 mmoles) was dissolved in H20
(20 mL) and cooled to 0°C. The 4- Butylbenzoyl chloride in dichloromethane was
added and triethylamine (4.2 mL, 30 mmoles) and the reaction was stirred over night.
H20 (20 mL) was added and the reaction was extracted with dichloromethane (3 20
mL), the organic layer was washed with a pH 2 buffer (3 * 20 mL), dried over Na2S0 4
and reduced in vacuo. The product was purified by recrystallisation from chloroform
and cold petroleum ether and washed with boiling ethyl acetate to give the product as a
white solid 1.44 g (50 %); p 204-205 ° C; dH (400 MHz, CDC1 ) 7.76 (d, 2H, J 8.5,
CH-C-'Bu), 7.66 (d, 1H, J 6, NH-CH), 7.42 (d, 2H, J 8.5, CH-C-CO), 6.00 (br.s, 1H,
NH-Cl), 4.67 (ddd, 1H, J 11, 6, 1.5, CH-C4), 3.34-3.19 (m, 2H, HI), 2.18 (br.d, IH, J
13, H4 equatorial), 2.00 (br.d, IH, J 12.5, H3 equatorial), 1.92-1.78 ( , 2H, H2
equatorial & H3 axial), 1.51 (q, IH, J 13, H4 axial), 1.33 (br.q, IH, J 11, H2 axial),
and 1.29 (s, 3H, C(CH3) ) ; C (100 MHz, CDC13) 176.0 (C=0), 166.2 (C=0), 155.0
(C-C(CH 3)3), 131.4 (C-C=0), 127.9, 125.4 (CH phenyl), 52.5 (CH-NH), 42.1 (CI),
34.9 (C(CH ) ), 3 1.6 (C4), 31.2 (C(CH 3)3) and 28.9, 28.0 (C2, C3); ESI m/z 100 %,
44
P66854. OO Spec as filed 8.06. 11
598.8 (M2Na+) and 32 %, 289.2 (MH+); HR ESI m/z (Ci 7H24N20 2Na requires
3 1 1.1730) found 3 11.1 736; ra /cm ' : 3210 (NH indole), 2906 (C-H), 1640 (C=0) and
1566 (aromatic); [a] D (c = 0.523, CHC13) +63.77.
Example 20: (S)-3-(4'-iert-Butylbenzoylamino)-tetrahydropyridin-2-one
(5 )-3-amino-tetrahydropyridin-2-one (33 mmoles) was dissolved in H20 (100 mL) and
cooled to 0°C. 4-tButylbenzoyl chloride (20 mmoles) in dichloromethane was added
and triethylamine (6.3 mL, 45 mmoles) and the reaction was stirred over night. The
reaction was extracted with dichloromethane (3 * 20 mL), the organic layer was
washed with a pH 2 buffer (3 20 mL), dried over Na2S0 and reduced in vacuo. The
product was purified by silica column chromatography (petroleum ether:ethyl
acetate:methanol 50:50:0 to 0:80:20) to give the product as a white solid 3.13 g (53 %)
p 195-196 °C; dH (400 MHz, CDC1 ) 7.24 (d, 2H, J 8.5, CH-C-¾u), 7.49 (d, IH, J 6,
NH-CH), 7.36 (d, 2H, J 8.5, CH-C-CO), 6.94 (br.s, H, NH-Cl), 4.62 (dt, 1H, J 11, 6,
1.5, CH-C4), 3.31-3.26 ( , 2H, HI), 2.52 (ddt, IH, J 13, 6, 4.5, H3 equatorial), 1.90-
183 ( , 2H, H2), 1.63 (tt, IH, J 12.5, 8.5, H3 axial) and 1.27 (s, 9H, C(CH )3); C (100
MHz, CDC13) 172.3 ( =0 ), 167.4 ( =0 ), 154.9 (C-C(CH )3), 131 .2 ( -C ), 127.0,
126.7, 125.3 ( H phenyl), 50.8 (CH-NH), 41.6 (CI), 34.9 (C(CH )3), 31.2 (C(CH 3)3),
27.2 (C3) and 2 1. 1 (C2); ESI m/z 100 %, 297.2 (MNa +) and 38 %, 275.2 (MH+);
ma m : 3251 (NH), 2959 (C-H), 1683, 1648 (C=0) and 1558 (aromatic); HR ESI
m/z (Ci 6H2 N20 2 requires 275.1754) found 275.1752; [a] 2
D (c = 0.515, CHCI3)
+82.52.
Example 21: (R)-3-(4'-feri-Butylbenzoylamino)-tetrahydropyridin-2-one
45
P668S4.WO0l .Spec as filed 8.06. 1
(fl)-3-amino-tetrahydropyridin-2-one (15 mmoles) was dissolved in H20 (100 mL) and
cooled to 0°C. 4- Butylbenzoyl chloride (10 mmoles) in dichloromethane was added
and triethylamine (4.2 mL, 30 mmoles) and the reaction was stirred over night. The
reaction was extracted with dichloromethane (3 20 mL), the organic layer was
washed with a pH 2 buffer (3 * 20 mL), dried over Na2S0 4 and reduced in vacuo. The
product was purified by silica column chromatography (petroleum ether:ethyl
acetate:methanol 50:50:0 to 0:80:20) to give the product as a white solid 1.03 g (38 %)
mp 193-194 °C; dH (400 MHz, CDC13) 7.72 (d, 2H, J 8.5, CH-C-'Bu), 7.49 (d, 1H, J 6,
NH-CH), 7.36 (d, 2H, J 8.5, CH-C-CO), 6.93 (br.s, 1H, NH-Cl), 4.39 (dt, 1H, J 11.5,
6, CH-C4), 3.31-3.26 (m, 2H, HI), 2.52 (ddt, 1H, J 12.5, 5.5, 4.5, H3 equatorial), 1.90-
1.87 ( , 2H, H2), 1.63 (tt, 1H, J 12.5, 8.5, H3) and 1.27 (s, 9H, C(CH )3) ; C (100
MHz, CDCI3) 172.3 (CO), 167.4 (0=0), 154.9 (C-C(CH ) ), 131.2 (C-C=0), 127.0,
125.3 ( H phenyl), 50.8 (CH-NH), 4 1.6 (CI), 34.9 (C(CH 3) ), 31.2 (C( H3)3), 27.2
(C3) and 21.1 (C2); /cm 1: 3247 (NH), 2958 (C-H), 1682, 1647 (C=0) and 1544
(aromatic); ESI m/z 19 %, 297.2 (MNa+) and 13 %, 275.2 (MH+); HR ESI m/z
(C H2 N20 2 requires 297.1573) found 275.1750; [a] D(c = 0.512, CHC1 ) -84.08.
Example 22: (S)-3-(4'-HexyIbenzoylamino)-azepan-2-one
(S)-3-amino-azepan-2-one hydrochloride (0.95 g, 5.79 mmoles) was dissolved in H20
(15 mL) and cooled to 0°C. 4-Hexylbenzoyl chloride (1.2 mL, 5 mmoles) in
46
P668S4.WOO Spec as filed 8.06. 11
dichloromethane was added and triethylamine (2.1 mL, 15 mmoles) and the reaction
was stirred over night. H20 (20 mL) was added and the reaction was extracted with
dichloromethane (3 x 20 mL), the organic layer was washed with a pH 2 buffer (3
mL), dried over Na S0 4 and reduced in vacuo. The product was purified by silica
column chromatography (petroleum ethenethyl acetate 50:50:0 to 0:80:20) to give the
product as a white solid 0.76 g (48 %); mp 167-168 ° C; d (400 MHz, CDC1 ) 7.74 (d,
2H, J 8, CH-C-Hex), 7.61 (d, 1H, J 6, NH-CH), 7.21 (d, 2H, J 8, C -C-CO), 6.54
(br.t, 1H, J 6, NH-C1), 4.69 (ddd, 1H, J 11, 6, 1.5, CH-C4), 3.37-3.22 (m, 2H, HI),
2.62 (t, 2H, J 7.5, H5), 2.21 (d, IH, J 13, H4 equatorial), 2.03 (dt, 1H, J 14, 3.5, H3
equatorial), 1.95-1.82 (m, 2H, H2 equatorial & H3 axial), 1.64-1.49 (m, 3H, H4 axial
& H6), 1.41 (q, IH, J 13, H2 axial) 1.33-1.23 ( , 6H, H7, H8 & H9) and 0.86 (t, 3H, J
7, H10); c (100 MHz, CDC1 ) 175.9 (C=0 lactam), 166.3 (C=0 amide), 147.0 (CHex),
131.6 (C-C=0), 128.5 (CH phenyl), 127.1 (CH phenyl), 52.6 (CH-NH), 42.2
(CI), 35.8 (C5), 31.7 (C4), 31.2 (C6), 29.0, 28.9, 28.0, (C2, C3, C7, and C8), 22.6
(C9) and 14.1 (CIO); /cm : 3244 (NH), 2956 (C-H), 1658, 1644 (C=0) and 1543
(aromatic); ESI m/z 43 %, 317.2 (MH+), 6% 339.2 (MNa+) and 6 %, 654.7 (M2Na+);
HR ESI m/z (C19H2 N20 2Na requires 339.2043) found 339.2050; [a] 25 (c = 0.507,
CHC1 ) +60.06.
Example 23: (S)-3-(4'-Hexylbenzoylamino)-tetrahydropyridin-2-one
(5)-3-amino-tetrahydropyridin-2-one (10 mmoles) was dissolved in H20 (35 mL) and
cooled to 0°C. 4-Hexylbenzoyl chloride (1.2 mL, 5 mmoles) in dichloromethane was
added and triethylamine (2. 1 mL, 15 mmoles) and the reaction was stirred over night.
The reaction was extracted with dichloromethane (3 x 15 mL), the organic layer was
washed with a pH 2 buffer (3 15 mL), dried over Na2S0 4 and reduced in vacuo. The
product was purified by silica column chromatography (petroleum ether:ethyl acetate
50:50:0 to 0:80:20) to give the product as a white solid 0.95 g (63 %); mp 118-1 19 0 C;
47
P66854.WO01 .Spec as filed 8.06. 1
d (400 MHz, CDCI3) 7.70 (d, 2H, J 8, CH-C-Hex), 7.38 (d, 1H, J 6, NH-CH), 7.17 (d,
2H, J 8, CH-C-CO), 6.81 (br.s, 1H, NH-C1), 4.39 (dt, 1H, J 11.5, 6, CH-C3), 3.33-
3.26 (m, 2H, HI), 2.58 (t, 2H, J 7.5, H4), 2.57-2.52 (m, 1H, H3 equatorial obscured by
H4), 1.92-1.84 (m, 2H, H2), 1.67-1.52 (m, 3H, H3 axial & H5), 1.29-1.23 (m, 6H, H6,
H7 & H8) and 0.84 (t, 3H, J 7.5, H9); ); C (100 MHz, CDCI3) 172.3 (C=0 lactam),
167.5 (C=0 amide), 146.9 (C-Hex), 131.5 (C-C=0), 128.4 (CH phenyl), 127.2 (CH
phenyl), 50.8 (CH-NH), 41.6 (CI), 35.8 (C4), 31.7 (C3), 31.2 (C5), 28.9 (C6), 27.2,
(C7), 22.6 (C8), 2 1. 1 (C2) and 14.1 (C9); cm : 3338, 3247 (NH), 2921 (C-H),
1656, 1637 (C=0) and 1562 (aromatic); ESI /z 100 %, 325.2 (MNa+) and 37% 303.2
(MH+); HR ESI m/z (Ci 8H26N20 2Na requires 325.1886) found 325.1883; [a] D (c =
0.51 1, CHClj) +79.55.
Example 24: (R)-3-(4 ,-HexylbenzoyIamino)-tetrahydropyridin-2-one
(i?)-3-amino-tetrahydropyridin-2-one (10 mmoles) was dissolved in H20 (20 L) and
cooled to 0°C. 4-Hexylbenzoyl chloride (1.2 mL, 5 mmoles) in dichloromethane was
added and triethylamine (2.1 mL, 15 mmoles) and the reaction was stirred over night.
The reaction was extracted with dichloromethane (3 15 mL), the organic layer was
washed with a pH 2 buffer (3 15 mL), dried over Na2S0 4 and reduced in vacuo. The
product was purified by silica column chromatography (petroleum ether: ethyl acetate
50:50:0 to 0:80:20) to give the product as a white solid 0.53 g (35 %); mp 117-1 18 C;
dH (400 MHz, CDCI3) 7.71 (d, 2H, J 8, CH-C-Hex), 7.28 (d, 1H, J 5, NH-CH), 7.19 (d,
2H, J 8, CH-C-CO), 6.52 (br.s, 1H, NH-C1), 4.41 (dt, 1H, J 11.5, 5.5, CH-C3), 3.35-
3.31 (m, 2H, HI), 2.67-2.60 (m, 1H, H3 equatorial), 2.61 (t, 2H, J 7.5, H4), 2.00-1 .89
(m, 2H, H2), 1.67-1.54 (m, 3H, H3 equatorial & H5), 1.32-1 .23 ( , 6H, H6, H7 & H8)
and 0.85 (t, 3H, J 7, H9); ); C (100 MHz, CDC13) 172.1 (C=0 lactam), 167.6 (C=0
amide), 147.0 (C-Hex), 131 .5 (C-C=0), 128.5 (CH phenyl), 127.2 (CH phenyl), 50.9
(CH-NH), 4 1.7 (CI), 35.8 (C4), 31.7 (C3), 31.2 (C5), 28.9 (C6), 27.2, (C7), 22.6 (C8),
48
P66854.WO01 .Spec as filed 8.06. 1
21.1 (C2) and 14.1 (C9); /cm 1: 3328, 3240 (NH), 2954 (C-H), 1651, 1635 (C=0)
and 1533 (aromatic); ESI m/z 100 %, 325.2 (MNa+) and 33% 303.2 ( H+); HR ESI
m/z (C, H26N20 Na requires 325.1886) found 325.1888; [ ] D (c = 0.496, CHC13) -
8 1.17.
Example 25: (S)-3-(4'-Octylbenzoylamino)-azepan-2-one
(5)-3-amino-azepan-2-one hydrochloride (0.91 g, 5.55 mmoles) was dissolved in H20
(15 mL) and cooled to 0°C. The 4-octylbenzoyl chloride in dichloromethane was
added and triethylamine (0.84 mL, 6 mmoles) and the reaction was stirred over night.
H20 (20 mL) was added and the reaction was extracted with dichloromethane (3 20
mL), the organic layer was washed with a pH 2 buffer (3 15 mL), dried over Na2S0 4
and reduced in vacuo. The product was purified by silica column chromatography
(petroleum ether:ethyl acetate 50:50:0 to 0:80:20) to give the product as a white solid
0.087 g (4 %); mp 159-160 ° C; dH (400 MHz, CDC13) 7.73 (d, 2H, J 8, CH-C-Oct),
7.64 (d, 1H, J 5.5, NH-CH), 7.20 (d, 2H, J 8, CH-C-CO), 6.94 (br.t, 1H, J 6, NH-C1),
4.68 (dd, 1H, J 1 , 6, CH-C4), 3.35-3.20 (m, 2H, HI), 2.60 (t, 2H, J 7.5, H5), 2.19 (d,
1H, J 13, H4 equatorial), 2.00 (br.d, 1H, H3 equatorial), 1.92-1.79 (m, 3H, H2 & H3
axial), 1.62-1.46 ( , 3H, H4 axial & H6), 1.38 (q, 1H, J 11.5, H2 axial) 1.30-1.19 (m,
10H, H7, H8, H9, H10 & H l ) and 0.84 (t, 3H, J 7.5, H12); C (100 MHz, CDC1 )
176.0 (C=0 lactam), 166.3 (C=0 amide), 146.9 (C-Oct), 131.6 (C-C=0), 128.5 (CH
phenyl), 127.1 (CH phenyl), 52.5 (CH-NH), 42.1 (CI), 35.8 (C5), 31.9 (C4), 3 1.6
(C6), .2 (C7), 29.4, 29.3, 28.9, 28.0, (C2, C3, C8, C9 and CIO), 22.7 (CI 1) and 14.1
(C12); w /cm 1 : 3204 (NH indole), 2923 (C-H), 1637 (amide C=0) and 1544
(aromatic); ESI m/z 100 %, 345.2 (MH+) and 9 %, 710.8 (M2Na+) ; HR ESI m/z
(C2 H32N20 2Na requires 367.2356) found 367.2361; [c ] D (c = 0.124, CDC13) +68.01.
49
P66854.WOO Spec as filed 8.06.1 1
Example 26: (S)-3-(4'-OctyIbenzoylamino)-tetrahydropyridin-2-one
2
(5)-3-amino-tetrahydropyridin-2-one (10 mmoles) was dissolved in H20 (35 mL) and
cooled to 0°C. 4-Octylbenzoyl chloride (2 mmoles) in dichloromethane was added and
triethylamine (0.85 mL, 6 mmoles) and the reaction was stirred over night. The
reaction was extracted with dichloromethane (3 10 mL), the organic layer was
washed with a pH 2 buffer (3 10 mL), dried over Na2S0 4 and reduced in vacuo. The
product was purified by silica column chromatography (petroleum ethenethyl acetate
50:50:0 to 0:80:20) to give the product as a white solid 0.55 g (83 %); p 122-123 ° C;
dH (400 MHz, CDC1 ) 7.71 (d, 2H, J 8, CH-C-Oct), 7.26 (d, 1H, J 5.5, NH-CH), 7.19
(d, 2H, J 8, CH-C-CO), 6.47 (br.s, 1H, NH-C1), 4.41 (dt, 1H, J 11.5, 5.5, CH-C3),
3.33-3.27 (m, 2H, HI), 2.84-2.58 ( , 1H, J 3, H3 equatorial obscured by H4), 2.60 (t,
2H, J 7.5, H4), 1.97-1.90 (m, 2H, H2), 1.67-1.54 ( , 3H, H3 axial & H5), 1.29-1.23
(m, 10H, H6, H7, H8, H9 & H10) and 0.86 (t, 3H, J 7, H 1); C (100 MHz, CDC13)
172.1 (C=0 lactam), 167.6 (C=0 amide), 147.0 (C-Oct), 131.5 (C-C=0), 128.5 (CH
phenyl), 127.2 (CH phenyl), 50.9 (CH-NH), 41.7 (CI), 35.8 (C4), 31.9 (C5), 31.2
(C3), 29.4 (C6), 29.3 (C7, C8), 27.2 (C9), 22.3 (CIO), 21.1 (C2) and 14.1 (CI 1);
ma /c : 3240 (NH), 2921 (C-H), 1653, 1615 (C=0) and 1563 (aromatic); ESI m/z
100 %, 353.2 (MNa+) and 47 %, 331 .2 (MH+); HR ESI m/z (C20H30N2O2Na requires
353.2199) found 353.2198; [a] D(c = 0.509, CHC13) +75.74.
Example 27: (R)-3-(4'-Octylbenzoylamino)-tetrahydropyridin-2-one
2
50
P66854 . OOl .Spec as filed 8.06. 11
(/?)-3-amino-tetrahydropyridin-2-one (5 mmoles) was dissolved in H 0 (25 mL) and
cooled to 0°C. 4-Octyl benzoyl chloride (2 mmoles) in dichloromethane was added and
triethylamine (0.85 mL, 6 mmoles) and the reaction was stirred over night. The
reaction was extracted with dichloromethane (3 * 10 mL), the organic layer was
washed with a pH 2 buffer (3 < 10 mL), dried over Na2S0 and reduced in vacuo. The
product was purified by silica column chromatography (petroleum ethenethyl acetate
50:50:0 to 0:80:20) to give the product as a white solid 0.16 g (25 %); mp 122-123 ° C;
dH (400 MHz, CDC13) 7.71 (d, 2H, J 8.5, C/f-C-Oct), 7.23 (d, 1H, J 6.5, N -CH), 7.20
(d, 2H, J 8.5, CH-C-CO), 6.34 (br.s, 1H, NH-C1), 4.41 (dt, 1H, J 11.5, 5.5, CH-C3),
3.37-3.33 (m, 2H, HI), 2.68 (ddt, 1H, J 13, 5.5, 4.5, H3 equatorial), 2.61 (t, 2H, J 7.5,
H4), 1.98-1 .91 (m, 2H, H2), 1.67-1.55 (m, 3H, H3 axial & H5), 1.31-1 .22 (m, 10H,
H6, H7, H8, H9 & H10) and 0.86 (t, 3H, J 7, H 1); C (100 MHz, CDC1 ) 172.1 (C=0
lactam), 167.6 (C=0 amide), 147.0 (C-Oct), 131.5 (C-C=0), 128.5 (CH phenyl), 127.2
(CH phenyl), 51.0 (CH-NH), 41.7 (CI), 35.8 (C4), 3 1.9 (C5), 31.2 (C3), 29.4 (C6),
29.3 (C7, C8), 27.2 (C9), 22.7 (CIO), 21.1 (C2) and 14.1 (Cl l); ESI m/z 39 %, 353.2
(M a+), 19 %, 331.2 (MH+) and 14 %, 682.7 (M2Na+); HR ESI m/z (C 0H 0N2O2H+
requires 331.2380) found 331.2381 ; cm : 3250 (NH), 2955 (C-H), 1653 (C=0)
and 1540 (aromatic); [a] (c = 0.485, CHC13) -77.80.
Pharmacological study of the products of the invention
A. Inhibition of MCP-1 induced leukocyte migration
Assay principle
The biological activity of the compounds of the current invention may be
demonstrated using any of a broad range of functional assays of leukocyte migration in
vitro, including but not limited to Boyden chamber and related transwell migration
assays, under-agarose migration assays and direct visualisation chambers such as the
Dunn Chamber.
5 1
P66854.WO01 .Spec as filed 8.06. 1
For example, to demonstrate the inhibition of leukocyte migration in response to
chemokines (but not other chemoattractants) the 96-well format micro transwell assay
system from Neuroprobe (Gaithersburg, MD, USA) has been used. In principle, this
assay consists of two chambers separated by a porous membrane. The chemoattractant
is placed in the lower compartment and the cells are placed in the upper compartment.
After incubation for a period at 37°C the cells move towards the chemoattractant, and
the number of cells in the lower compartment is proportional to the chemoattractant
activity (relative to a series of controls).
This assay can be used with a range of different leukocyte populations. For example,
freshly prepared human peripheral blood leukocytes may used. Alternatively,
leukocyte subsets may be prepared, including polymorphonuclear cells or lymphocytes
or monocytes using methods well known to those skilled in the art such as density
gradient centrifugation or magnetic bead separations. Alternatively, immortal cell
lines which have been extensively validated as models of human peripheral blood
leukocytes may be used, including, but not limited to THP-1 cells as a model of
monocytes or Jurkat cells as model of naive T cells.
Although a range of conditions for the assay are acceptible to demonstrate the
inhibition of chemokine-induced leukocyte migration, a specific example is hereby
provided.
Materials
The transwell migration systems are manufactured by Neuroprobe, Gaithersburg, MD,
USA.
The plates used are ChemoTx plates (Neuroprobe 101-8) and 30 mΐ clear plates
(Neuroprobe MP30).
Geys' Balanced Salt Solution is purchased from Sigma (Sigma G-9779).
Fatty acid-free BSA is purchased from Sigma (Sigma A-8806).
52
P66854.WO0l .Spec as filed 8.06. 1
TT, i.e. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, is purchased
from Sigma (Sigma M-5655).
RPMI-1640 without phenol red is purchased from Sigma (Sigma R-8755).
The THP-1 cell line (European Cell culture Collection) were used as the leukocyte cell
population.
Testprotocol
The following procedure is used for testing the invention compounds for MCP- 1
induced leukocyte migration:
First, the cell suspension to be placed in the upper compartment is prepared. The THP-
1 cells are pelleted by centrifugation (770 x g; 4 mins) and washed with Geys
Balanced Salt Solution with g ml BSA (GBSS + BSA). This wash is then repeated,
and the cells repelleted before being resuspended in a small volume of GBSS + BSA
for counting, for example using a standard haemocytometer.
The volume of GBSS + BSA is then adjusted depending on the number of cells present
so that the cells are at final density of 4.45 x 106 cells per ml of GBSS + BSA. This
ensures that there are 100,000 THP-1 cells in each 25m1of the solution that will be
placed in the upper chamber of the plate.
To test a single compound for its ability to inhibit MCP-1 induced migration, it is
necessary to prepare two lots of cells. The suspension of THP-1 cells at 4.45 x 106
cells/ml is divided into two pots. To one pot the inhibitor under test is added at an
appropriate final concentration, in an appropriate vehicle (for example at 1mM in not
more than 1% DMSO). To the second pot an equal volume of GBSS + BSA plus
vehicle as appropriate (e.g. not more than 1% DMSO) is added to act as a control.
Next, the chemoattractant solution to be placed in the lower compartment is prepared.
MCP-1 is diluted in GBSS + BSA to give a final concentration of 25 ng/ml. This is
divided into two pots, as for the cell suspension. To one pot, the test compound is
added to the same final concentration as was added to the cell suspension, while to the
53
P6685 4.WOO Spec as filed 8.06.1 1
other pot an equal volume of GBSS + BSA plus vehicle as appropriate (e,g. not more
than 1% DMSO) is added.
Note that the volume of liquid that needs to be added to make the addition of the text
compound needs to be taken into account, when establishing the final concentration of
MCP-1 in the solution for the lower compartment and the final concentration of cells
in the upper compartment.
Once the chemoattractant solutions for the lower wells and cell solutions for the upper
chambers have been prepared, the migration chamber should be assembled. Place
29 mΐ of the appropriate chemoattractant solution into the lower well of the chamber.
Assays should be performed with at least triplicate determinations of each condition.
Once all the lower chambers have been filled, apply the prous membrane to the
chamber in accordance with the manufacturer's instructions. Finally, apply 25 m of
the appropriate cell solution to each upper chamber. A plastic lid is placed over the
entire apparatus to prevent evaporation.
The assembled chamber is incubated at 37 °C, 5% C0 2, for 2 hours. A suspension of
cells in GBSS + BSA is also incubated under identical conditions in a tube: these cells
will be used to construct a standard curve for determining the number of cells that have
migrated to the lower chamber under each condition.
At the end of the incubation, the liquid cell suspension is gently removed from the
upper chamber, and 20m1of ice-cold 20mM EDTA in PBS is added to the upper
chamber, and the apparatus is incubated at 4°C for 15 mins. This procedure causes any
cells adhering to the underside of the membrane to fall into the lower chamber.
After this incubation the filter is carefully flushed with GBSS + BSA to wash off the
EDTA, and then the filter is removed.
The number of cells migrated into the lower chamber under each condition can then be
determined by a number of methods, including direct counting, labelling with
fluorescent or radioactive markers or through the use of a vital dye. Typically, we
utilise the vital dye MTT. 3 mΐ of stock MTT solution are added to each well, and then
the plate is incubated at 37 °C for 1-2 hours during which time dehydrogenase
54
P668S4. OO Spec as filed 8.06.1 1
enzymes within the cells convert the soluble MTT to an insoluble blue formazan
product that can be quantified spectrophotometrically.
In parallel, an 8-point standard curve is set up. Starting with the number of cells added
to each upper chamber (100,000) and going down in 2-fold serial dilutions in GBSS +
BSA, the cells are added to a plate in 25 mΐ , with 3 m of MTT stock solution added.
The standard curve plate is incubated along side the migration plate.
At the end of this incubation, the liquid is carefully removed from the lower chambers,
taking care not to disturb the precipitated formazan product. After allowing to air dry
briefly, 20m1of DMSO is added to each lower chamber to solubilise the blue dye, and
absorbance at 595nm is determined using a 96-well plate reader. The absorbance of
each well is then interpolated to the standard curve to estimate the number of cells in
each lower chamber.
The MCP-1 stimulated migration is determined by subtracting the average number of
cells that reached the lower compartment in wells where no MCP-1 was added from
the average number of cells that reached the lower compartment where MCP-1 was
present at 25ng/ml.
The impact of the test substance is calculated by comparing the MCP-1 -induced
migration which occurred in the presence or absence of various concentrations of the
test substance. Typically, the inhibition of migration is expressed as a percentage of
the total MCP-1 induced migration which was blocked by the presence of the
compound. For most compounds, a dose-response graph is constructed by determining
the inhibition of MCP-1 induced migration which occurs at a range of different
compound concentrations (typically ranging from I M to 1mM or higher in the case of
poorly active compounds). The inhibitory activity of each compound is then expressed
as the concentration of compound required to reduce the MCP-1-induced migration by
50% (the ED50 concentration).
Results
The compounds of reference examples 1 to 14 were tested and were shown to have an
ED50 of 100 nM or less in this test.
55
P 6854.WOO Spec as filed 8.06. 1
B. In vivo assay
The anti-inflammatory efficacy of an exemplary compound according to the present
invention was tested using the murine sub-lethal endotoxemia model. This model has
been widely used to demonstrate the anti-inflammatory effect of compounds in vivo -
Fox et 2009, J Med Chem. 52(1 1): 3591-3595.
Briefly, the method is as follows: Female CD1 mice (28-30g, ~7 weeks of age) were
dosed with their respective treatment in sterile filtered 1% CMC by oral gavage in a
dose volume of lOml kg one hour prior to an endotoxin (LPS) challenge. The
endotoxin challenge was injected by the intraperitoneal route containing 675,000
Endotoxin Units of LPS (E. coli strain 0 111:B4 (Code L4130)) in endotoxin free PBS.
Mice were left for two hours and then exsanguinated under terminal anaesthesia and
blood was taken. Serum was prepared from this terminal bleed and aliquoted and
stored at -20°C. Serum TNF-a levels were measured by ELISA per manufacturers
instructions (R and D Systems).
Eight animals were treated in each group, and the data for the animal with the highest
and lowest TNF- a level in each group were eliminated, and the mean and standard
error reported for the remaining six animals. Data for untreated animals were taken
from an historical control experiment.
A single dose of (S)-4-Fluoro-N-(2-oxopiperidin-3-yl)benzamide (also known as (S)-
3-(4'-fluorobenzoylamino)-tetrahydropyridin-2-one; see Example 3
above) administered by oral gavage, inhibited endotoxin-stimulated TNF-alpha levels
by 50% (see Fig. 2, column B).
This experiment demonstrates that the compounds according to the invention have
anti-inflammatory activity in vivo.
56
P66854.WO01 .Spec as filed 8.06. 11

Claims
1. A compound of general formula (I), or a pharmaceutically acceptable salt thereof,
for use in the treatment of an inflammatory disorder:
(I)
wherein
n is an integer from 1 to 4;
k is an integer from 0 to 5, representing the number of groups substituting C2, C , C4,
C5 and/or C6 of the benzyl ring; and
X are linear or branched groups substituting the benzyl ring independently selected
from any one of the group consisting of: alkyl, haloalkyl, hydroxyalkyl, hydroxy,
alkoxy, amino, aminoalkyl, aminodialkyl, carboxy, and halogen;
with the proviso that:
when on the benzyl ring C2, C5 and C are unsubstituted, and C4 is unsubstituted or is
substituted with an hydroxy, alkoxy, amino, aminoalkyl, aminodialkyl, or halogen
group, then C is substituted with a halogen group; and
when on the benzyl ring C , C5 and C6 are unsubstituted, and C3 is unsubstituted or is
substituted with an alkyl, haloalkyl, hydroxyalkyl, hydroxy, alkoxy, amino,
aminoalkyl, aminodialkyl or carboxy group, then C4 is substituted with any one of the
group consisting of: alkyl group, haloalkyl group, hydroxyalkyl group, and carboxy
group.
57
P66854. WOOl Spec as filed 8.06. 11
2. A compound of formula ( '), or a pharmaceutically acceptable salt thereof, for use
in the treatment of an inflammatory disorder:
wherein n, k and X are defined as in claim 1.
3. Use of a compound of general formula (I), or a pharmaceutically acceptable salt
thereof, in the manufacture of a medicament for the treatment of an inflammatory
disorder:
(I)
wherein
n is an integer from 1 to 4;
k is an integer from 0 to 5, representing the number of groups substituting C2, C , C4,
C and/or C of the benzyl ring; and
X are linear or branched groups substituting the benzyl ring independently selected
from any one of the group consisting of: alkyl, haloalkyl, hydroxyalkyl, hydroxy,
alkoxy, amino, aminoalkyl, aminodialkyl, carboxy, and halogen;
with the proviso that:
58
P66854.WO01 .Spec as filed 8.06.1 1
when on the benzyl ring C , C5 and C are unsubstituted, and C4 is unsubstituted or is
substituted with an hydroxy, alkoxy, amino, aminoalkyl, aminodialkyl, or halogen
group, then C is substituted with a halogen group; and
when on the benzyl ring C , C5 and C are unsubstituted, and C is unsubstituted or is
substituted with an alkyl, haloalkyl, hydroxyalkyl, hydroxy, alkoxy, amino,
aminoalkyl, aminodialkyl or carboxy group, then C4 is substituted with any one of the
group consisting of: alkyl group, haloalkyl group, hydroxyalkyl group and carboxy
group.
4. Use of a compound of formula (F), or a pharmaceutically acceptable salt thereof,
the manufacture of a medicament for the treatment of an inflammatory disorder:
( )
wherein n, k and X are defined as in claim 3.
5. A compound of general formula (I):
(I)
wherein
n is an integer from 1 to 4;
59
P66854.WO0l .Spec as filed 8.06.1 1
k is an integer from 1 to 5, representing the number of groups substituting C2, C3, C ,
C5 and/or C of the benzyl ring;
when n is 1 or 2, X are linear or branched groups independently selected from any one
of the group consisting of: C7 or higher alkyl, haloalkyl with a C7or higher ky group,
hydroxyalkyl with a C7 or higher alkyl group, C7or greater alkoxy, aminoalkyl with a C4 or
higher alkyl group, aminodialkyl with two C4or higher alkyl groups, and carboxy;
when n is 3 or 4, X are linear or branched groups independently selected from any one
of the group consisting of: alkyl, haloalkyl, hydroxyalkyl, hydroxy, alkoxy, amino,
aminoalkyl, aminodialkyl, carboxy, and halogen;
with the proviso that:
when n is 3 or 4 and on the benzyl ring C2, C5 and C are unsubstituted, and C4 is
unsubstituted or is substituted with an hydroxy, alkoxy, amino, aminoalkyl,
aminodialkyl, or halogen group, then C3 is substituted with a halogen group;
when n is 3 or 4 and on the benzyl ring C2, C5 and C are unsubstituted, and C3 is
unsubstituted or is substituted with an alkyl, haloalkyl, hydroxyalkyl, hydroxy, alkoxy,
amino, aminoalkyl, aminodialkyl or carboxy group, then C4 is substituted with any one
of the group consisting of: alkyl group, haloalkyl group, hydroxyalkyl group, and
carboxy group; and
when
provided that the compound is none of the group consisting of: 3-(3 -
trifluoromethylbenzoylamino)-caprolactam, 3-(4'-methylbenzoylamino)-caprolactam,
3-(2'-aminobenzoylamino)-caprolactam, 3-(3',4'-dimethoxybenzoylamino)-
caprolactam, 3-(3',5'-di-t rt-butyl -4'- hydroxybenzoylamino)-caprolactam, 3-(2',4'-
dimethoxybenzoylamino)-caprolactam, 3-(3 '-methoxybenzoylamino)-caprolactam, 3-
(4'-trifluoromethylbenzoylamino)-caprolactam, 3-(2',3',4'-trimethoxybenzoylamino)-
caprolactam, 3-(2',6'-difluoromethylbenzoylamino)-caprolactam, 3-(2'-
fluoromethylbenzoylamino)-caprolactam, 3-(2'-amino-3'-hydroxy-4'-
methylbenzoylamino)-caprolactam, and 3-(3 ',5 ' -dimethylbenzoylamino)-caprolactam.
60
P66854.WO01 .Spec as filed 8.06. 11
6. A compound of formula (G ) :
')
wherein n, k and X are defined as in either of claim 1 or claim 5,
provided that the compound is compound is none of the group consisting of: (S)-3-(4'-
methoxybenzoylamino)-caprolactam, (S)-3-(4'-methylbenzoylamino)-caprolactam,
(S)-3-(3'-trifluoromethylbenzoylamino)-caprolactam, (S)-3-(2'-carboxybenzoylamino)-
caprolactam, and (S)-3-(3', 4',5'-trimethoxybenzoylamino)-caprolactam.
7. A pharmaceutical composition comprising, as active ingredient, a compound as
defined in either of claims 5 or 6, or a pharmaceutically acceptable salt thereof, and at
least one pharmaceutically acceptable excipient and/or carrier.
8. The compound, use or composition according to any preceding claim, wherein n=2.
9. The compound, use or composition according to any preceding claim, wherein n=3.
10. The compound, use or composition according to any preceding claim, wherein X is
haloalkyl, for example trifluoromethyl.
11. The compound according to claims 1 or 2, or the use according to claims 3 or 4,
wherein the compound is selected from the group consisting of:
(S)-3-(4'-methylbenzoylamino)-caprolactam, and
(S)-3-(3',5'-dimethylbenzoylamino)-caprolactam,
and pharmaceutically acceptable salts thereof.
12. The compound according to any of claims 1, 2 or 6, or the use according to claims
3 or 4, wherein the compound is selected from the group consisting of:
6 1
P66854. OO Spec as filed 8.06.1
(S)-3-Fluoro-N-(2-oxopiperidin-3-yl)benzamide,
(5)-2-Fluoro-N-(2-oxopiperidin-3-yl)benzamide,
(5)-4-Fluoro-N-(2-oxopiperidin-3-yl)benzamide,
(5)-N-(2-Oxopiperidin-3-yl)-4-(trifluoromethyl)benzamide,
(S)-N-(2-Oxopiperidin-3 -yl)-3 -(trifluoromethyl)benzamide,
(S)-N-(2-Oxopiperidin-3-yl)-2-(trifluoromethyl)benzamide,
(.S)-2,3-difluoro-N-(2-oxopiperidin-3-yl)benzamide,
(5)-2,4-difluoro-N-(2-oxopiperidin-3-yl)benzamide,
(-y)-2,5-difluoro-N-(2-oxopiperidin-3-yl)benzamide,
(S)-2,6-difluoro-N-(2-oxopiperidin-3-yl)benzamide,
(S)-3,4-difluoro-N-(2-oxopiperidin-3-yl)benzamide,
(5)-3,5-difluoro-N-(2-oxopiperidin-3-yl)benzamide,
(S)-3-(3'-Butylbenzoylamino)-azepan-2-one,
(S)-3-(4'-Ethylbenzoylamino)-tetrahydropyridin-2-one,
(S)-3-(4'-Butylbenzoylamino)-tetrahydropyridin-2-one,
(S)-3-(4'-tert-Butylbenzoylamino)-tetrahydropyridin-2-one, and
(S)-3-(4'-Hexylbenzoylamino)-tetrahydropyridin-2-one,
and pharmaceutically acceptable salts thereof.
13. The compound according to any of claims , 2, 5 or 6, or the use according to
claims 3 or 4, wherein the compound is selected from the group consisting of:
(S)-3-(4'-Ethylbenzoylamino)-azepan-2-one,
(S)-3-(4'-Butylbenzoylamino)-azepan-2-one,
(S)-3-(4'-t r t-Butylbenzoylamino)-azepan-2-one,
(S)-3-(4'-Hexylbenzoylamino)-azepan-2-one,
(S)-3-(4'-Octylbenzoylamino)-azepan-2-one, and
(S)-3-(4'-Octylbenzoylamino)-tetrahydropyridin-2-one,
and pharmaceutically acceptable salts thereof.
14. The compound according to claim 1, or the use according to claim 3, wherein the
compound is selected from the group consisting of:
62
P66854. WOOLSpec as filed 8.06.1 1
(R)-3-(4'-Butylbenzoylamino)-tetrahydropyridin-2-one,
(R)-3-(4 '-t rt-Butylbenzoylamino)-tetrahydropyridin-2-one, and
(R)-3-(4'-Hexylbenzoylamino)-tetrahydropyridin-2-one,
and pharmaceutically acceptable salts thereof.
15. The compound according to either of claim 1 or claim 5, or the use according to
claim 3, wherein the compound is (R)-3-(4'-Octylbenzoylamino)-tetrahydropyridin-2-
one or a pharmaceutically acceptable salt thereof.
16. The compound according to either of claims 1 or 2 or the use according to either
of claims 3 or 4, wherein the inflammatory disorder is selected from the group
consisting of autoimmune diseases, asthma, rheumatoid arthritis, a disorder
characterised by an elevated TNF- level, psoriasis, allergies, multiple sclerosis,
fibrosis (including diabetic nephropathy), and formation of adhesions.
17. The compound or use according to claim 16, wherein the inflammatory disorder is
formation of adhesions.
18. The compound or use according to claim 6 or claim 17, wherein the compound is
administered locally.
19. A method of treatment, amelioration or prophylaxis of the symptoms of an
inflammatory disease (including an adverse inflammatory reaction to any agent) by the
administration to a patient of an anti-inflammatory amount of a compound,
pharmaceutical composition or medicament as defined in any preceding claim.
20. A library consisting of elements all of which have structures according to the
formula (I) or ( ) as defined in any of claims lto 18, and hence which all have anti¬
inflammatory activity, useful for screening compounds for novel or improved
properties in a particular assay of anti-inflammatory activity.