Note: Descriptions are shown in the official language in which they were submitted.
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MODULATORS OF ALPHA7 NICOTINIC ACETYLCHOLINE
RECEPTORS AND THERAPEUTIC USES THEREOF
The present invention relates to compounds with ca7 nicotinic
acetylcholine receptor (ca7 nAChR) agonistic activity, processes for their
preparation, pharmaceutical compositions containing the same and the use
thereof for the treatment of neurological and psychiatric diseases.
Background of the invention
A number of recent observations point to a potential neuroprotective
effect of nicotine in a variety of neurodegeneration models in animals and in
cultured cells, involving excitotoxic insults (1-5), trophic deprivation (6),
ischemia (7), trauma (8), A13-mediated neuronal death (9-11) and protein-
aggregation mediated neuronal degeneration (9;12). In many instances where
nicotine displays a neuroprotective effect, a direct involvement of receptors
comprising the ca7 subtype has been invoked (7;11-15) suggesting that
activation of cx7 subtype-containing nicotinic acetylcholine receptors may be
instrumental in mediating the neuroprotective effects of nicotine. The
available
data suggest that the a7 nicotinic acetylcholine receptor represents a valid
molecular target for the development of agonists/positive modulators active as
neuroprotective molecules. Indeed, ca7 nicotinic receptor agonists have
already
been identified and evaluated as possible leads for the development of
neuroprotective drugs (16-20). Involvement of ca7 nicotinic acetylcholine
receptor in inflammatory processes has also recently been described (21).
Thus,
the development of novel modulators of this receptor should lead to novel
treatments of neurological, psychiatric and inflammatory diseases.
Known prior art
Different compounds cariying an aryl/heteroaryl- ureido or carbamoyl
moiety and a basic nitrogen and exhibiting nicotinic and muscarinic
CONFIRMATION COPY
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acetylcholine receptor affinity or claimed for use in Alzheimer disease were
found to be described: fused pyridazine derivatives (W003070707); methods
for treating an inflammatory condition (US2004127536); aza-bicyclic N-
biarylamides (W003078431); heterocyclic urea derivatives (W00266468);
spirocyclic piperidines (W02004004714); phenyl-substituted indoles and
indazoles (WO0174773); phenyl-subsituted imidazopyridines (WO0174815);
Heterocyclic compounds carrying a basic nitrogen and hexibiting
various types of biological activity were found to be described: anti herpes
virus compounds (US6288091); 2H-phthalazin-l-one derivatives
(W000044726); 1,4-dihydro-2(2H)isoquinolines (DE2406490); pyridine
compounds (JP06016638); piperidine amides (WO0198268); 8-amino-aryl-
substituted imidazopyrazines as kinase inhibitors (US2005009832);
Heterocyclic compounds were also disclosed in Heterocycles (1997),
45(4), 723-734: 1-[w [(arylamino)carbonyl]alkyl]-4-
(benzocycloalkyl)piperazines.
These referenced compounds are readily distinguishable structurally
from those herein disclosed by either the functionalities, attachment chain or
substitution pattern; these prior documents fail to disclose or suggest the
unique combination of structural fragments which embody the novel
w-aminoalkylureas or carboxamides having activity on the nicotinic alpha7
receptor herein disclosed.
Summary of the invention
The invention provides novel compounds acting as full or partial
agonists at the a7 nicotinic acetylcholine receptor (a7 nAChR),
pharmaceutical compositions containing the same compounds and the use
thereof for the treatment of diseases that may benefit from the activation of
the alpha 7 nicotinic acetylcholine receptor such as neurological and
psychiatric disorders, in particular Alzheimer's disease and schizophrenia.
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Description of the invention
This invention provides compounds of formula (I)
Ki
X I
~ ()W/ ( I)k",, ()n /Y,,, Q/ (R')i
K2 I
R
(I)
wherein
w, h and k are, independently from one another, 0,1,2, or 3 with the
condition that 3<_w + h + k<_5;
Kl and K2, which are bound to either the same or a different carbon
atom where k>1, represent independently from one another hydrogen;
halogen; (CI-C5) alkyl, alkoxy, fluoroalkyl, alkylene, fluoroalkylene;
hydroxyalkyl; or K1 and K2 taken together may form an alkylidene or a
fluoroalkylidene group; or K1 and K2, taken together with the carbon atom to
which they are attached, form a (C3-C6) cycloalkyl group; or when k is 2,
two Ok carbon atoms may form an unsaturated bond; or when w is 1, 2, or 3,
and k is 1, K1 and K2 taken together with the carbon atom to which they are
attached may form an oxo group;
j is 0, 1 or 2;
X is a group of formula
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T-P
Oq LZ T~~N N~T'P = U
/N-()q' N-j n ~-N.u,
T-P T'P N
4)S I~-Ps T'P4S
.N On N On On
T'p T-P
N 4)S ~ ~ S
()n N ()n
Z is CH2, N, 0, S, S(=0), or S(=0)2;
p is 0, 1, 2 or 3;
nis0,1or2;
s is 1 or 2;
q and q' are, independently from one another, integers from 1 to 4;
T' represent, independently from one another, hydroxy; mercapto;
amino; cyano; nitro; linear, branched or cyclic (C1-C6) alkyl, trihaloalkyl,
hydroxyalkyl, aminoalkyl, mercaptoalkyl, alkoxy, alkylthio, alkylcarbonyl,
alkoxycarbonyl, alkylcarbonylamino; mono- or di-, linear, branched or cyclic
(C1-C6) alkylamino; linear, branched or cyclic (C1-C6) alkoxy-(C1-C6) alkyl,
mono- or di- (C1-C6) alkylamino-(C1-C6) alkyl, or (C1-C6) alkylthio-(CI-C6)
alkyl; (C1-C3) alkylsulphonylamino; mono- or di- (C1-C3)
alkylaminosulphonyl; sulphamoyl; linear, branched or cyclic (CI-C6)
alkylaminocarbonyl; carbamoyl; or, when p is 2 or 3, two T' form a 5- to 8-
membered ring with spiro or fused junction;
U and U' represent, independently from one another, hydrogen; cyano;
hydroxy; amino; a mono- or di-, linear, branched, or cyclic (C1-C6) alkylamino
group; a linear or branched (CI-C6) alkoxy group; a linear, branched or cyclic
(C1-C6) alkyl, azaalkyl, oxaalkyl chain optionally substituted with hydroxy,
mercapto, amino, cyano, nitro, oxo, trihalomethyl, trihalomethoxy, carbamoyl,
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sulphamoyl, linear, branched or cyclic (C1-C6) alkyl, hydroxyalkyl,
aminoalkyl, mercaptoalkyl, alkoxy, alkylthio, alkylcarbonyl, alkoxycarbonyl,
alkylcarbonylamino, mono- or di-, linear, branched or cyclic (CI-C6)
alkylaminocarbonyl, mono- or di- (C5-Clo) aryl- or heteroarylaminocarbonyl,
5 (C5-Cto) aryl- or heteroarylsulphonylamino, (C1-C3) alkylsulphonylamino,
(C5-CIo) aryl- or heteroarylsulphonyl, (CI-C3) alkylsulphonyl, mono- or
di- (C5-CJo) aryl- or heteroarylsulphamoyl, mono- or di- (CI-C3)
alkylsulphamoyl, mono- or di-, linear, branched, or cyclic (CI-C6) alkylamino;
a linear, branched or cyclic (CI-C6) alkyl, azaalkyl, oxaalkyl chain bearing a
5- to 10-membered aryl or heteroaryl group optionally substituted with one or
more groups independently selected from hydroxy, mercapto, amino, cyano,
nitro, trihalomethyl, trihalomethoxy, linear, branched or cyclic (C1-C6)
alkyl,
hydroxyalkyl, aminoalkyl, mercaptoalkyl, alkoxy, alkylthio, alkylcarbonyl,
alkoxyxcarbonyl, alkylcarbonylamino, mono- or di-, linear, branched, or
cyclic (CI-C6) alkylamino, linear, branched or cyclic (CI-C6) alkoxy-(C1-C6)
alkyl, mono- or di- (CI-C6) alkylamino-(C1-C6) alkyl, or (C1-C6) alkylthio-
(CI-C6) alkyl, carbamoyl, (C5-CIo) aryl- or heteroarylsulphonylamino, (C1-C3)
alkylsulphonylamino, mono- or di- (C5-Clo) aryl- or heteroarylsulphamoyl,
(CI-C3) alkylsulphamoyl, sulphamoyl, mono- or di-, linear, branched or cyclic
(CI-C6) alkylaminocarbonyl; mono- or di- (C5-Clo) aryl- or
heteroarylaminocarbonyl, a 5 to 10 membered aromatic or heteroaromatic ring
optionally substituted with one or more groups independently selected from
hydroxy; halogen; mercapto; amino; cyano; nitro; trihalomethyl;
trihalomethoxy; linear, branched or cyclic (CI-C6) alkyl, hydroxyalkyl,
aminoalkyl, mercaptoalkyl, alkoxy, alkylthio, alkylcarbonyl-,
alkoxyxcarbonyl, alkylcarbonylamino; mono- or di-, linear, branched, or
cyclic (C1-C6) alkylamino; linear, branched or cyclic (CI-C6) alkoxy-(C1-C6)
alkyl, mono- or di- (CI-C6) alkylamino-(CI-C6) alkyl, or (C1-C6) alkylthio-
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(CI-C6) alkyl; carbamoyl; (C5-CIo) aryl- or heteroarylsulphonylamino; (C1-C3)
alkylsulphonylamino; mono- or di- (C5-CIo) aryl- or heteroarylsulphamoyl;
mono- or di- (C1-C3) alkylsulphamoyl; sulphamoyl; mono- or di- (C5-Clo)
aryl- or heteroarylaminocarbonyl; mono- or di-, linear, branched or cyclic
(C1-C6) alkylaminocarbonyl;
-Y-Q- is -C(=O)NH-Q- or -NH-C(=O)-NH-Q;
Q is a 5 to 10-membered aromatic or heteroaromatic ring;
R represents a 5 to 10-membered aromatic or heteroaromatic ring,
optionally substituted with one or more groups independently selected from:
halogen; hydroxy; mercapto; cyano; nitro; amino; linear, branched or cyclic
(CI-C6) alkyl, trihaloalkyl, alkoxy or alkylcarbonyl; linear, branched, or
cyclic
(CI-C6) alkylcarbonylamino, mono- or di- (C5-CIO) aryl- or
heteroarylaminocarbonyl; mono- or di, linear, branched, or cyclic (C1-C6)
alkylaminocarbonyl; carbamoyl; linear, branched, or cyclic (CI-C6)
alkylsulphonylamino; linear, branched, or cyclic (C1-C6) alkylsulphonyl;
mono- or di- (C5-Clo) aryl- or heteroarylsulphamoyl; mono- or di- linear,
branched, or cyclic (C1-C6) alkylsulphamoyl; linear, branched or cyclic
(CI-C6) alkoxy-(CI-C6) alkyl, mono- or di- (CI-C6) alkylamino-(C1-C6) alkyl,
(CI-C6) alkylthio-(CI-C6) alkyl;
R' represent, independently from one another when j= 2, halogen;
hydroxy; mercapto; cyano; nitro; trihalomethyl; trihalomethoxy; linear,
branched or cyclic (CI-C6) alkyl, trihaloalkyl, alkoxy, hydroxyalkyl,
mercaptoalkyl, alkoxycarbonyl, alkylcarbonyl, alkylsulphonyl; linear,
branched, or cyclic (C1-C6) alkylcarbonylamino; mono- or di, linear, branched,
or cyclic (CI-C6) alkylaminocarbonyl; carbamoyl; linear, branched, or cyclic
(CI-C6) alkylsulphamoyl; linear, branched or cyclic (C1-C6) alkoxy-(CI-C6)
alkyl, mono- or di- (C1-C6) alkylamino-(Cl-C6) alkyl, (CI-C6) alkylthio-
(C I-C6) alkyl,
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provided that when k is zero and the sum of w and h is 4, T' is mercapto;
amino; trihaloalkyl; hydroxyalkyl; (C 1-C6) aminoalkyl; mercaptoalkyl;
alkylthio; alkoxycarbonyl; alkylcarbonylamino; mono- or di-, linear, branched
or cyclic (C1-C6) alkylamino; linear, branched or cyclic (C1-C6) alkoxy-(C1-
C6)
alkyl, mono- or di- (C1-C6) alkylamino-(C1-C6) alkyl, or (CI-C6) alkylthio-(CI-
C6) alkyl; mono- or di- (C1-C3) alkylaminosulphonyl, or j74;
and that when j is zero, and the sum of w, h and k is 4, then K1 and K2
are not both hydrogen;
and with the exclusion of the following compounds:
1--[4-(2-Amino-thiazol-5-yl)-phenyl]-3-(3-imidazol-1-yl-propyl)-
urea;1-(Biphenyl-4-yl)-3-(5-(spiro(indane-1,4'-piperidine-10-yl)-pentyl)-urea;
1-(Biphenyl-4-yl)-3-(4-(spiro(indane- 1,4'-piperidine-10-yl)-butyl)-urea;3-{4-
[3-(3-Morpholin-4-yl-propyl)-ureido]-phenyl}-1H-indazole-5-carboxylic acid
amide;3- {4-[3-(3-Piperidin-1-yl-propyl)-ureido]-phenyl } -1 H-indazole-5-
carboxylic acid amide;l-[4-(8-Methylamino-imidazo[1,2-a]pyrazin-3-yl)-
phenyl]-3 -(3-morpholin-4-yl-propyl)-urea
1-[4-(8-Cyclopropylamino-imidazo[ 1,2-a]pyrazin-3-yl)-phenyl]-3-(3-
pyrrolidin-1-yl-propyl)-urea; l -(2-Hydroxy-3-morpholin-4-yl-propyl)-3-[4-(8-
methylamino-imidazo[ 1,2-a]pyrazin-3-yl)-phenyl]-urea; l-[4-(8-
Cyclopropylamino-imidazo[ 1,2-a]pyrazin-3-yl)-phenyl]-3-(3-morpholin-4-yl-
propyl)-urea; l-[4-(8-Methylamino-imidazo[ 1,2-a]pyrazin-3-yl)-phenyl]-3-(3-
pyrrolidin-l-yl-propyl)-urea; N-Biphenyl-4-yl-4-piperazin-l-yl-butyramide
Within compounds of formula I above described, most preferred
compounds are those in which X is
T'p
()q~z U
I I N
NOQ' ;
/ `U'
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More preferred group compounds are those for which X is
T'p
OqLz
I I
/N-Oq'
In one embodiment, this invention provides compounds hereafter
referred to as G1, in which:
X is:
T'p
1)qLz T'p U
I I , N ~N,N,
N-Oq N
z is selected from CH2, N, 0;
T' represent, independently from one another when p is greater than 1,
hydroxy; amino; cyano; nitro; linear, branched or cyclic (C1-C6) alkyl,
trihaloalkyl, hydroxyalkyl, aminoalkyl, mercaptoalkyl, alkoxy, alkylthio,
alkylcarbonyl, alkoxycarbonyl, alkylcarbonylamino; mono- or di-, linear,
branched or cyclic (C1-C6) alkylamino; linear, branched or cyclic (C1-C6)
alkoxy-(CI-C6) alkyl, mono- or di- (CI-C6) alkylamino-(CI-C6) alkyl, or
(CI-C6) alkylthio-(CI-C6) alkyl; (C1-C3) alkylsulphonylamino; mono- or
di- (C1-C3) alkylaminosulphonyl; sulphamoyl; linear, branched or cyclic
(Cl-C6) alkylaminocarbonyl; carbamoyl; or when p is 2 or 3, two T'
substituents form a 5- to 8-membered ring with spiro or fused junction;
U and U' represent, independently from one another, hydrogen; a linear,
branched or cyclic (C I-C6) alkyl, azaalkyl, oxaalkyl chain optionally
substituted with hydroxy, oxo, trihalomethyl, trihalomethoxy, carbamoyl,
sulphamoyl, pyridyl, linear, branched or cyclic (CI-C3) alkylcarbonyl,
alkoxycarbonyl, alkylcarbonylamino, mono- or di-, linear, branched or cyclic
(C1-C3) alkylaminocarbonyl, (CI-C3) alkylsulphonylamino, (CI-C3)
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alkylsulphonyl, mono- or di- (C1-C3) alkylsulphamoyl, mono- or di-, linear,
branched, or cyclic (C I-C6) alkylamino;
Q is a 6 to 10-membered aromatic or heteroaromatic ring;
R represents a 5 to 10-membered aromatic or heteroaromatic ring optionally
substituted with one or more groups independently selected from: halogen;
hydroxy; mercapto; cyano; nitro; amino; linear, branched or cyclic (C1-C6)
alkyl,
trihaloalkyl,. alkoxy or alkylcarbonyl; linear, branched, or cyclic (C1-C6)
alkylcarbonylamino; mono- or di, linear, branched, or cyclic (C1-C6)
alkylaminocarbonyl; carbamoyl; linear, branched, or cyclic (C1-C6)
alkylsulphonylamino; linear, branched, or cyclic (C1-C6) alkylsulphonyl; mono-
or
di- linear, branched, or cyclic (C1-C6) alkylsulphamoyl; linear, branched or
cyclic
(C1-C6) alkoxy-(C1-C6) alkyl, mono- or di- (CI-C6) alkylamino-(C1-C6) alkyl;
R' represent, independently from one another when j = 2, halogen;
hydroxy; trihalomethyl; trihalomethoxy; linear, branched or cyclic (C1-C3)
alkyl, trihaloalkyl, alkoxy, hydroxyalkyl, alkoxycarbonyl, alkylcarbonyl,
alkylsulphonyl; linear, branched, or cyclic (C1-C3) alkylcarbonylamino; mono-
or di, linear, branched, or cyclic (C1-C3) alkylaminocarbonyl; carbamoyl;
(CI-C3) alkylsulphonylamino; linear, branched, or cyclic (C1-C3)
alkylsulphamoyl; linear, branched or cyclic (CI-C3) alkoxy-(Cf-C3) alkyl,
mono- or di- (CI-C3) alkylamino-(C1-C3) alkyl, (C1-C3) alkylthio-(C1-C3)
alkyl;
In one aspect of this invention, G 1 provides a group of compounds
hereafter referred to as G 1 a wherein w + h + k= 4.
Within G 1 a, a particular embodiment provides compounds in which
k is 0;
X is a group of formula:
U
iN,U.
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U and U' represent, independently from one another, hydrogen; a linear,
branched or cyclic (CI-C6) alkyl, azaalkyl, oxaalkyl chain optionally
substituted with trihalomethyl, trihalomethoxy, carbamoyl, sulphamoyl,
pyridyl;
5 -Y- is a group -C(=O)NH- or -NH-C(=O)NH-;
j is 0, or 1;
R represents a 5 to 10-membered aromatic or heteroaromatic ring,
optionally substituted with one or more groups independently selected from:
halogen; hydroxy; linear, branched or cyclic (C1-C6) alkyl, trihaloalkyl,
10 alkoxy; linear, branched, or cyclic (CI-C3) alkylcarbonylamino; mono- or
di,
linear, branched, or cyclic (CI-C3) alkylaminocarbonyl; carbamoyl;
R' represents halogen; trihalomethyl; trihalomethoxy; linear, branched
or cyclic (C1-C3) alkyl, alkoxy.
In another embodiment, this invention provides a group of compounds
hereafter referred to as G2 in which:
w+h+k=4
K1 and K2 represent, independently from one another hydrogen;
halogen; (CI-C3) alkyl, alkoxy;
X is a group of formula:
Tp
OqLz
I I
N-()4'
z is CH2, N, 0;
p is 0 or 1;
T' represents linear, branched or cyclic (C1-C3) alkyl, trihaloalkyl,
alkylcarbonyl, alkoxycarbonyl, alkylcarbonylamino; linear, branched or cyclic
(C1-C3) alkylaminocarbonyl; carbamoyl;
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R represents a 5 to 10-membered aromatic or heteroaromatic ring,
optionally substituted with one or more groups independently selected from:
halogen; hydroxy; linear, branched or cyclic (C1-C3) alkyl, trihaloalkyl,
alkoxy
or alkylcarbonyl; linear, branched, or cyclic (C1-C3) alkylcarbonylamino; mono-
or di, linear, branched, or cyclic (C1-C3) alkylaminocarbonyl; carbamoyl;
linear,
branched, or cyclic (C1-C3) alkylsulphonylamino; linear, branched, or cyclic
(C1-C6) alkylsulphonyl; mono- or di- linear, branched, or cyclic (C1-C6)
alkylsulphamoyl; linear, branched or cyclic (C1-C6) alkoxy-(C1-C6) alkyl,
mono- or di- (CI-C6) alkylamino-(C1-C6) alkyl;
R' represent, independently of one another when j = 2, halogen; hydroxy;
trihalomethyl; trihalomethoxy; linear, branched or cyclic (C1-C3) alkyl,
trihaloalkyl, alkoxy; linear, branched, or cyclic (CI-C3) alkylcarbonylamino;
mono- or di, linear, branched, or cyclic (C1-C3) alkylaminocarbonyl;
carbamoyl;
(CI-C3) alkylsulphonylamino; linear, branched, or cyclic (C1-C3)
alkylsulphamoyl; linear, branched or cyclic (CI-C3) alkoxy-(C1-C3) alkyl, mono-
or di- (C1-C3) alkylamino-(CI-C3) alkyl, (C1-C3) alkylthio-(C1-C3) alkyl;
Within G2, a particular embodiment defines a group of compounds
hereafter referred to as G3 in which:
K 1 and K2 represent, independently from one another hydrogen;
halogen; (CI-C3) alkyl;
X is a group of formula:
()4 I
~p
N-Oq'
z is CH2, N;
q and q' are, independently from one another, integers from 1 to 3;
T' represents linear, branched or cyclic (CI-C3) alkyl, alkylcarbonyl;
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R represents a 5 to 10-membered aromatic or heteroaromatic ring,
optionally substituted with one or more groups independently selected from:
halogen; linear, branched or cyclic (CI-C3) alkyl, trihaloalkyl, alkoxy;
linear,
branched, or cyclic (CI -C3) alkylcarbonylamino; mono- or di, linear,
branched,
or cyclic (C1-C3) alkylaminocarbonyl; carbamoyl;
R' represents, independently from one another when j = 2, halogen;
trihalomethyl; linear, branched or cyclic (CI-C3) alkyl, alkoxy.
In one aspect, G3 provides a subset of compounds hereafter referred to
as G4a in which -Y- is a group -C(=O)NH-.
Within G4a, a particular embodiment is that in which:
Q is a phenyl or pyridyl ring;
j is 1 or 2;
R represents a phenyl, pyridyl, or pyrazolyl ring, optionally substituted
with one or more groups independently selected from: halogen; linear,
branched or cyclic (CI-C3) alkyl, trihaloalkyl, alkoxy; linear, branched, or
cyclic (CI-C3) alkylcarbonylamino; mono- or di, linear, branched, or cyclic
(C1-C3) alkylaminocarbonyl; carbamoyl.
In another aspect, G3 provides a sub-set of compounds hereafter referred
to as G4b where:
-Y- is a group -NH-C(=0)-NH-.
Within G4b, a particular embodiment is that in which:
K1 and K2 represent, independently from one another hydrogen;
halogen; (CI-C3) alkyl;
X is a group of formula:
Tp
0qLz
I I
N-Oq'
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z is CH2, N;
pis0or1;
q and q' are, independently from one another, integers from 1 to 3;
T' represents linear, branched or cyclic (C1-C3) alkyl, alkylcarbonyl;
-Y- is a group -NH-C(=O)-NH-;
Q is a phenyl or pyridyl;
R represents a phenyl, pyridyl or pyrazole ring, optionally substituted
with one or more groups independently selected from: halogen; linear,
branched or cyclic (C1-C3) alkyl, trihaloalkyl, alkoxy; linear, branched, or
cyclic (CI-C3) alkylcarbonylamino; mono- or di, linear, branched, or cyclic
(C1-C3) alkylaminocarbonyl; carbamoyl;
Under yet another aspect of invention, G1 also provides a group of
compounds hereafter referred to as G5, wherein
w+ h+ k= 3;
X is
T'p
Oq-Lz
I I
N-()q'
Within G5, one embodiment provides a group of compounds hereafter
referred to as G5a in which:
q and q' are, independently from one another, integers from 1 to 3;
T' represent, independently from one another when p is greater than 1,
hydroxy; cyano; oxo; linear, branched or cyclic (C1-C6) alkyl, trihaloalkyl,
hydroxyalkyl, alkoxy, alkylcarbonyl, alkoxycarbonyl, alkylcarbonylamino;
linear, branched or cyclic (C1-C6) alkoxy-(C1-C6) alkyl; (C1-C3)
alkylsulphonylamino; mono- or di- (CI-C3) alkylaminosulphonyl; sulphamoyl;
linear, branched or cyclic (CI-C6) alkylaminocarbonyl; carbamoyl; linear,
branched or cyclic (CI-C3) alkoxy-(C1-C3) alkyl; (C1-C3)
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alkylsulphonylamino; mono- or di- (C1-C3) alkylsulphamoyl; (C1-C3)
sulphonyl;
-Y- is a group -NH-C(=O)-NH-;
R represents a 5 to 6-membered aromatic or heteroaromatic ring,
optionally substituted with one or more groups independently selected from:
halogen; hydroxy; cyano; linear, branched or cyclic (C1-C3) alkyl,
trihaloalkyl,
alkoxy; linear, branched, or cyclic (CI-C3) alkylcarbonylamino; mono- or di,
linear, branched, or cyclic (C1-C3) alkylaminocarbonyl; carbamoyl; linear,
branched, or cyclic (C1-C3) alkylsulphonylamino; linear, branched, or cyclic
(CI-C3) alkylsulphonyl; mono- or di- linear, branched, or cyclic (CI-C3)
alkylsulphamoyl; linear, branched or cyclic (C1-C3) alkoxy-(C1-C3) alkyl;
R' represents, independently of one another when j = 2, halogen;
trihalomethyl; trihalomethoxy; linear, branched or cyclic (CI-C3) alkyl,
alkoxy.
Within G5a, a particular embodiment is represented by compounds in
which
k is 0;
p is 0, or 1;
T' represents, independently from one another when p is greater than 1,
linear, branched or cyclic (C1-C3) alkyl, alkylcarbonyl, alkoxycarbonyl,
alkylcarbonylamino; linear, branched or cyclic (CI-C3) alkylaminocarbonyl;
carbamoyl;
-Y- is a group -NH-C(=O)-NH-;
Q is a phenyl or pyridyl;
j is 0 or 1;
R represents a phenyl or pyridyl ring optionally substituted with one or
more groups independently selected from: halogen; hydroxy; linear, branched
or cyclic (C1-C6) alkyl, alkoxy; linear, branched, or cyclic (C1-C3)
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alkylcarbonylamino; mono- or di, linear, branched, or cyclic (C1-C3)
alkylaminocarbonyl; carbamoyl;
R' represents halogen.
The compounds of the invention can be prepared through a number of
5 synthetic routes amongst which the ones illustrated in Schemes 1-5 below:
a) Scheme 1
r
HO
HO,~iNHz + oR Y OR
2 3 4
Y'= isocyanate Y = -NHCONH-
~ / + X NaBH(OAc)3 ~Y - Y ~
Y I\ R R
O X X\__
' R
5 6
la ~R
According to Scheme 1, an aminoalcohol, for example 3-amino-2,2-
10 dimethyl-propan-l-ol when k = 1 and K1 = K2 = -CH3, is reacted with for
example an isocyanate or a carbamoyl chloride, hereby exemplified by an
arylisocyanate, in an organic solvent such as for example dichloromethane,
tetrahydrofuran, dimethylformamide or mixtures thereof, until the reaction is
complete. The hydroxyurea 3 then oxidised under standard conditions (for
15 example Swern oxidation) and the obtained aldehyde 4 is then reacted with a
suitably substituted amine 6 under standard reductive alkylation conditions -
for example with sodium triacetoxyborohydride - to afford compound Ia. In
the case of R being a halogen or a boronic acid ester, Ia can be further
processed - for example via a cross-coupling reaction, for example under the
conditions of the Suzuki coupling, with a boronic acid or an aryl or
heteroaryl
halide - to yield compounds Io.
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b) Scheme 2
NHZ _
k ~ Y X
Br
Br~/Ok~ + K Q
CI 1J-R i K,I
7 8 9
X Kn /4)k Y X K~ Kk Y X= amine
+ K2 ~ ' ~ R Y = -CONH
I R (,
/
IR
l(x
According to Scheme 2 an a)-haloalkanoyl chloride is reacted with an
(hetero)aromatic amine 8 in the presence of an organic base to afford an
co-haloalkanoic acid amide 9. This species is reacted with an amine 6 to
displace the halogen and afford compounds Ia. In the case of R being a
halogen or a boronic acid ester, Ia can be further processed - for example via
a
cross-coupling reaction, for example under the conditions of the Suzuki
coupling, with a boronic acid or an aryl or heteroaryl halide - to yield
compounds I0.
c) Scheme 3
K, O ~ K O K,
Br,,OW O I ~OhN \ / + X X~OW O I I - , Oh,N ? ~ -~ X"OW O Oh,NHZ
Kz 0 Kz O Kz
, 0 1' 1'
X~ OW O i~ ~ NHZ + X~~)W0 i~ ~)h _N 0 X" OW0 i~Oh _N O
Oh Kz y ~R.O.(R')1 HN", iR HN~ R
O Oi
Ia 1 10 1
(R')j (R')1
According to Scheme 3, a suitably activated co-haloalkylphthalimide, is
reacted with an amine X in an organic solvent such as for example but not
limited to 2-butanone or dimethylformamide in the presence of a base such as
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for example triethylamine or potassium carbonate. For example, a mixture of
amine (or its hydrochloride salt) and w-haloalkylphthalimide are refluxed in
methylethyl ketone in the presence of alkaline carbonate until the reaction is
complete, then the reaction mixture is cooled, the insoluble materials removed
by filtration, the filtrate washed with chloroform or dichloromethane, and the
filtrate and washings concentrated to dryness. In the following step, the
co-aminoalkylphthalimide is converted into a w-diamine, for example by
refluxing a mixture of the w-aminoalkylphthalimide and hydrazine hydrate in
ethanol. The w-diamine is reacted with an activated species such as for
example an isocyanate or a carbamoyl chloride in an organic solvent such as
dichloromethane, tetrahydrofuran, dimethylformamide or mixtures thereof, to
give compounds of formula Ia In the case of R being a halogen, a boronic acid
or a boronic ester, Ia can be further processed - for example via a
cross-coupling reaction, for example under the conditions of the Suzuki
coupling, with a boronic acid or an aryl or heteroaryl halide - to yield
compounds of formula I.
d) Scheme 4
K, I' H I ' H
O
Q~W Ok , NHZ + II -~ Q~ Ow-( ~~ ()h .N` 'Q -~ Q"OW ( ~~()h
O ,Ny
Oh N If~lj Kz R.Q, (R')1 HN~Q-IR HN~Q/R
I I
K (R')1 i i (R')1
amine X I' H
reducing agent x,, OW.Ok~ ,N Q x~OW~O i~Oh'N Q
~ Oh y ~~
/~ (1 /R (R')l ~Q~R
Ia (R')1 ~a
According to Scheme 4, a suitable aldehyde precursor, for example an
aminoalcohol, for example aminopropanol when k = 1 and KI = K2 = H, is
reacted with for example an isocyanate or a carbamoyl chloride, hereby
exemplified by an arylisocyanate, in an organic solvent such as for example
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dichloromethane, tetrahydrofuran, dimethylformamide or mixtures thereof,
until the reaction is complete. The aldehyde precursor thus obtained is then
converted to the aldehyde, for example oxidised under standard conditions
(for example Swern oxidation) in the case of an alcohol, and the aldehyde is
then reacted with a suitably substituted amine X under standard reductive
alkylation conditions - for example with sodium triacetoxyborohydride - to
afford compounds Ia. In the case of R being a halogen, a boronic acid or a
boronic acid ester, Ia can be further processed - for example via a cross-
coupling reaction, for example under the conditions of the Suzuki coupling,
with a boronic acid or an aryl or heteroaryl halide - to yield compounds of
formula I.
e) Scheme 5
~, 0 ~, 0 amine X i,
0
Br~OW-0 i~) ~LG + R QH(R h - Br,O`N.0 i~() ~NH ba_~ x\OW O On _NH
K2 K2 R.Q=(R')j I(Z R"Q'(R')1
la
K,
1 0
0N I O ~NH
X\ 0 \
KZ R.Q=(R')J
Ip
According to Scheme 5 a suitably activated c)-haloalkanoic acid (for
example where the moiety C(=O)-LG represents an acyl chloride or activated
ester or imidazolide) is reacted with an (hetero)aromatic amine in the
presence
of an organic base to afford an w-haloalkanoic acid amide. This species is
further reacted with an amine X to displace the w-halogen atom and afford
compounds of formula Ia. In the case of R being a halogen, a boronic acid or a
boronic acid ester, Ia can be further processed - for example via a
cross-coupling reaction, for example under the conditions of the Suzuki
coupling, with a boronic acid or an aryl or heteroaryl halide - to yield
compounds of formula I.
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f) Scheme 6
K1 0 K
X ()k x 4R')1 1 ' O 4R')1
OW~ I\()ri OH + HzN.Q.R X~OW Ok\ ~N.Q.R
Kz I () H
X = amine K2
According to Scheme 6, an a)-aminoalkanoic acid is suitably activated
using an agent such for example but not limited to as 1,1'-carbonyldiimidazole
in a solvent such as for example dichloromethane, dimethylformamide or
mixtures thereof and reacted with a suitable heterocyclic amine to afford
subject matter compounds of formula I.
g) Scheme 7
Ki K
1 o
4R,)~ I 0 4R')1
X~OW O \ ~OH X
~()W.() i\()h HQ.Br
I () + HzN.Q.Br
Kz Kz
X = amine
K
1 ' O 4R')1
X"OW.() i \() ~H.Q.R
Kz
According to Scheme 7, an w-aminoalkanoic acid is suitably activated
using an agent such for example but not limited to as 1,1'-carbonyldiimidazole
in a solvent such as for example dichloromethane, dimethylformamide or
mixtures thereof and reacted with a suitable bromoaryl or heteroaryl amine to
afford bromoaryl or heteroaryl amides, which are then reacted further under
cross-coupling conditions, for example Suzuki conditions, to afford subject
matter compounds of formula I.
Scheme 8 shows one possible route towards the synthesis of chain-
substituted acids, precursors to compounds of Formula I
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O O 1) base ALK
O O~ O HBr 48%, 120 C O
2) a,w dihaloalkane Br O~ Br OH
n n
O O
A~ ALK
n=a2
0
0 amine X,
MeOH, H SO toluene, 0 NaOH aq X n OH
2 a Br n O reflux X n O ALK
ALK ALK
According to Scheme 8, an alkyl-substituted malonic acid diester it
treated with base, such as for example but not limited to sodium hydride in a
solvent such as tetrahydrofurane or dimethylformamide and reacted with an
5 a,w-dihaloalkane. The disubstituted malonic acid diester thus obtained is
hydrolysed and mono-decarboxylated by treatement with a strong acid, such
as for example hydrobromic acid. Esterification is then carried out, for
example by treatement with methanol and a catalytic amount of acid.
Substitution of the w-halogen may be accomplished by the use of a suitable
10 amine heating in a solvent like toluene, but not limited to this solvent.
Finally,
hydrolysis of the ester function with an aqueous base affords intermediates
which can be activated as described to afford compounds of Formula I.
The compounds in this invention can in general be prepared by any of
several standard synthetic processes commonly used by those skilled in the art
15 of organic chemistry. In general, the amides can be prepared through a
base-catalysed nucleophilic addition between the appropriate carboxylic acid
with an appropriately selected amine or via a nucleophilic substitution
reaction wherein the appropriate amine reacts with either the selected acyl
halide, anhydride or ester to yield the required amine. When coupling the
20 acids to the amines, standard chemical coupling reagents such as
carbonyldiimidazole (CDI), 1,3-dicyclohexylcarbodiimide (DCC) or 1-ethyl-
3-(3'-dimethylaminopropyl)carbodiimide hydrochloride (EDCI) can be used in
the presence or absence of hydroxybenzotriazole (HOBt). In an alternative
procedure the carboxylic acids are converted into the corresponding acyl
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halides by reaction with, for example, thionyl chloride or oxalyl chloride.
Subsequently, the acid halide is added to the appropriately selected amine to
yield the amide using art-known reaction procedures such as the Schotten-
Baumann method.
The carboxylic acids and the amines are readily available, or may be
prepared using methods that are well known in the art. Many compounds are
commercially available, for example, from Aldrich Chemicals, or when the
compounds are not commercially available, they may be readily prepared from
available precursors using straightforward transformations that are well known
in the art. For example the carboxylic acids can be prepared by hydrolysis of
nitriles, carbonation of organometallic compounds or oxidation of primary
alcoholds or aldehydes. In particular branched alkyl nitriles are prepared
from
the corresponding alkyl acetonitriles by conversion to the dialkyl or
spiroalkyl
derivative using e.g. sodium hexamethyldisilazane and methyl iodide or
dibromobutane, followed by hydrolysis under acidic or basic conditions to the
desired carboxylic acid. Appropriate acids and bases in the hydrolysis are for
example H2SO4 and KOH. The hydrolysis reaction can be conveniently
performed using microwave heating.
The compounds of formula I, their optical isomers or diastereomers can
be purified or separated according to well-known procedures, including but
not limited to chromatography with a chiral matrix and fractional
crystallisation.
The pharmacological activity of a representative group of compounds of
formula I was demonstrated in an in vitro assay utilising cells stably
transfected with the alpha 7 nicotinic acetylcholine receptor and cells
expressing the alpha 1 and alpha 3 nicotinic acetylcholine receptors and 5HT3
receptor as controls for selectivity. According to a further aspect, the
invention is therefore directed to a method of treating neurological and
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psychiatric disorders, which comprises administering to a subject, preferably
a
human subject in need thereof, an effective amount of a compound of formula
1. Neurological and psychiatric disorders that may benefit from the treatment
with the invention compounds include but are not limited to senile dementia,
attention deficit disorders, Alzheimer's disease and schizophrenia. In
general,
the compounds of formula I can be used for treating any disease condition,
disorder or dysfunction that may benefit from the activation of the alpha 7
nicotinic acetylcholine receptor, including but not limited to Parkinson's
disease, Huntington's chorea, amyotrophic lateral sclerosis, multiple
sclerosis,
epilepsy, memory or learning deficit, panic disorders, cognitive disorders,
depression, sepsis and arthritis.
The dosage of the compounds for use in therapy may vary depending
upon, for example, the administration route, the nature and severity of the
disease. In general, an acceptable pharmacological effect in humans may be
obtained with daily dosages ranging from 0.01 to 200 mg/kg.
In yet a further aspect, the invention refers to a pharmaceutical
composition containing one or more compounds of formula I, in association
with pharmaceutically acceptable carriers and excipients. The pharmaceutical
compositions can be in the form of solid, semi-solid or liquid preparations,
preferably in form of solutions, suspensions, powders, granules, tablets,
capsules, syrups, suppositories, aerosols or controlled delivery systems. The
compositions can be administered by a variety of routes, including oral,
transdermal, subcutaneous, intravenous, intramuscular, rectal and intranasal,
and are preferably formulated in unit dosage form, each dosage containing
from about 1 to about 1000 mg, preferably from 1 to 600 mg of the active
ingredient. The compounds of the invention can be in the form of free bases or
as acid addition salts, preferably salts with pharmaceutically acceptable
acids.
The invention also includes separated isomers and diastereomers of
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compounds I, or mixtures thereof (e.g. racemic mixtures). The principles and
methods for the preparation of pharmaceutical compositions are described for
example in Remington's Pharmaceutical Science, Mack Publishing Company,
Easton (PA).
Experimental Procedures - Synthesis of compounds
General
Unless otherwise specified all nuclear magnetic resonance spectra were
recorded using a Varian Mercury Plus 400 MHz spectrometer equipped with a
PFG ATB Broadband probe.
HPLC-MS analyses were performed with a Waters 2795 separation
module equipped with a Waters Micromass ZQ (ES ionisation) and Waters
PDA 2996, using a Waters XTerra MS C18 3.5 m 2.1x50mm column.
Preparative HLPC was run using a Waters 2767 system with a binary
Gradient Module Waters 2525 pump and coupled to a Waters Micromass ZQ
(ES) or Waters 2487 DAD, using a Supelco Discovery HS C18 5.0 m
10 x 21.2 mm column.
Gradients were run using 0.1% formic acid/water and 0.1% formic
acid/acetonitrile with gradient 5/95 to 95/5 in the run time indicated.
All column chromatography was performed following the method of
Still, C.; J. Org Chem 43, 2923 (1978). All TLC analyses were performed on
silica gel (Merck 60 F254) and spots revealed by UV visualisation at 254 nm
and KMnO4 or ninhydrin stain.
When specified for array synthesis, heating was performed on a Buchi
Syncore system.
All microwave reactions were performed in a CEM Discover oven.
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Abbreviations used throughout the Experimental Procedures
CDI N,N'-carbonyldiimidazole
DCM Dichloromethane
DCE 1,2-dichloroethane
DMEA N,N-dimethylethylamine
DMF N,N-dimethylformamide
DMSO, dmso Dimethylsulphoxide
DAM N,N-dimethylacetamide
SCX strong cation exchanger
TEA Triethylamine
TFA trifluoroacetic acid
THF Tetrahydrofuran
TLC thin layer chromatography
LC-MS, LCMS Liquid chromatography - mass spectrometry
HPLC High performance liquid chromatography
General procedure for aminoalkylamine synthesis - phthalimide route
N-(3-Bromopropyl)phthalimide (1 eq) was added to a suspension of
amine (1 eq), sodium iodide (0.5 eq) and potassium carbonate (1.1 eq) in
2-butanone (ca. 20 volumes with respect to phthalimide weight). The resulting
suspension was stirred for 18h at 85 C, then the reaction was filtered and the
solvent removed by vacuum distillation; the resulting oil was washed with
water and recovered with DCM. The solvent was removed under reduced
pressure to yield the amino-phthalimide products pure enough to be used in
the next step without further purification.
The phthalimides thus obtained (1 eq) were dissolved in EtOH (ca. 7-10
volumes with respect to phthalimide weight) and hydrazine monohydrate
(2 eq) was added dropwise. The mixture was heated at 80 C for 4 h, after
which the reaction was acidified with 37% HC1 and the solid which
precipitated was removed by filtration. The solution was concentrated under
vacuum and taken up with 1N HC1. Any residual 2,3-dihydro-phthalazine-1,4-
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dione was removed by filtration. The aqueous solution was evaporated under
vacuum to recover pure product.
In case of acid sensitive derivatives the reaction mixture was filtered
and washed with EtOH, concentrated under vacuum and taken up with toluene
5 and DCM to remove excess 2,3-dihydro-phthalazine-1,4-dione. Solvent
removal under reduced pressure afforded the pure product.
1 -[4-(3-Amino propyl) piperazin-1 ylJ-ethanone
2-[3-(4-Acetyl-piperazin-1-yl)-propyl]-isoindole-1,3-dione (5.0 g,
16 mmol) was dissolved in EtOH (50 mL) and hydrazine monohydrate
10 (1.5 mL, 32 mmol) was added. The mixture was heated at 80 C for 1 hour.
1- (4-Bromo phenyl)-3-(3 piperidin-1 yl propyl)-urea
a) 2-(3-Piperidin-1-yl propyl)-isoindole-1, 3-dione
N-(3-Bromopropyl)phthalimide (5.36 g, 20 mmol) was added to a
suspension of piperidine (1.98 mL, 20 mmol), sodium iodide (3.9 g, 26 mmol)
15 and potassium carbonate (4.15 g, 21 mmol) in 2-butanone (100 mL). The
resulting suspension was stirred for 24 hour at 90 C.
The reaction was filtered to remove inorganic salts and the solvent
removed by vacuum distillation; the resulting oil was washed with water and
recovered with DCM. The solvent was removed under reduced pressure to
20 afford 4.39 g of desired product as a white solid (yield: 81%).
C16H2ON202 Mass (calculated) [272.35],; found [M+H+]=273
'H-NMR (400 MHz, CDC13) 1.25-1.47(6H, m), 1.76-1.92 (2H, m),
2.14-2.42 (6H, m), 3.63-3.68 (2H, m), 7.63-7.73 (2H, m), 7.76-7.86 (2H, m).
b) 3-Piperidin-1 yl propylamine hydrochloride
25 2-(3-Piperidin-1-yl-propyl)-isoindole-1,3-dione (7.5 g, 18.34 mmol)
was dissolved in EtOH (45 mL) and hydrazine monohydrate (1.78 mL,
34.5 mmol) was added. The mixture was heated at 80 C for 1 hour. The
reaction was then cooled and acidified with 5 mL of 37% HC1 and
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2,3-dihydro-phthalazine-1,4-dione was removed by filtration. The solution
was concentrated under vacuum, 10 mL of 1N HC1 were added and the
mixture was filtered again to remove the residual 2,3-dihydro-phthalazine-1,4-
dione. The aqueous solution was evaporated under vacuum and 3.0 g (yield
48%) of pure product were recovered.
C8H18N2 Mass (calculated) [142.25]; (found) [M+H+]=143
NMR (400 MHz, dmso-d6): 1.25-1.45 (2 H, m), 1.57-1.92 (4 H, m),
1.93-2.17 (2H, m), 4.05-5.37 (4H, m).
General procedure for urea synthesis
To a cooled solution of amine (1 eq) in dichloromethane an equimolar
amount of an aryl or heteroaryl isocyanate was added. In the case of the amine
being in the form of a hydrochloride or bis-hydrochloride salt, equimolar
amounts of TEA were added to free-base the amine.
The mixture was left stirring at 0 C for 1-4 hours. The p-bromophenyl
ureas generally precipitated out of solution as white solids, were recovered
by
filtration and if necessary purified further by washing with Et20 or by flash
chromatography. The m-bromophenylureas were isolated by solvent removal
under reduced pressure and purified by crystallisation from an mixture of
AcOEt: EtZ0.
Alternatively, to a solution of aniline in DCM (0.32 mmol/mL) cooled
at 0 C, triphosgene (1 eq) was added under N2 flux; a white precipitate formed
and NEt3 (1.1 eq) was added and the mixture generally became a yellow
solution. After 30 minutes the amine was added and the reaction was left
stirring at 0 C for 2 hours, when LC-MS generally showed complete
conversion. If a precipitate was present it was filtered affording the urea
product, otherwise the solution was washed with NaOH 10% solution and the
organic phase concentrated under reduced pressure. The crude was then
purified by crystallization from Et20.
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1 -(4-Bromo phenyl)-3-(3 piperidin-1-yl propyl)-urea
To a cooled solution of 3-piperidin-1-yl-propylamine dihydrochloride
(0.96 g, 4 mmol) in dichloromethane (20 mL) TEA was added (1.11 mL,
8 mmol).
p-Bromophenylisocyanate (0.78 g, 4 mmol) was then added and the
mixture was stirred at 0 C until a white solid precipitated out of solution
after
2 hours. The white solid was filtered off and washed with Et20 to give 1.4 g
of pure title compound.
C 15H22BrN3OMass (calculated) [340.27]; (found) [M+H+]=340-342
Lc Rt (10 min method)= 2.69, 92%
NMR (400 MHz, dmso-d6): 1.26-1.63 (8H, m); 2.14-2.36 (4H, m);
2.45-2.51 (2H, m, under DMSO); 3.00-3.13 (2H, m); 6.11-6.26 (1 H, m), 7.33
(4H, s), 8.52-8.66(1H,s).
1- (2-Chloro-4-Bromo phenyl)-3-(4 piperidin-1 yl-butyl)-urea
Prepared via the general procedure for urea synthesis (via isocyanate)
Yield: 76%
NMR (400 MHz, dmso-d6): 1.27-1.52 (IOH, m), 2.14-2.37 (6H, m),
3.03-3.4 (2H, m), 7.04 (1 H, t), 7.40 (1 H, dd), 7.60 (1 H, d), 8.50 (1 H, s),
8.12
(1H, d).
1 -(2-Fluoro-4-Bromo phenyl)-3-(4 piperidin-1 yl-butyl)-urea
Prepared via the general procedure for urea synthesis (via isocyanate)
Yield: 88%
NMR (400 MHz, dmso-d6): 1.22-1.50 (IOH, m), 2.12-2.37 (6H, m),
3.00-3 .13 (2H, m), 6.62 (1 H, t), 7.25 (1H, d), 7.47 (1 H, dd), 8.10 (1H, t),
8.33
(1H, s).
1-(2, 6 difluoro-4-Bromo phenyl)-3-(4 piperidin-1 yl-butyl)-urea
Prepared via the general procedure for urea synthesis (via isocyanate)
Yield: 67%
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C16H22BrF2N3O Mass (calculated) 390; (found) [M+H+]= 390-392
Lc Rt= 2.04 (100%), 10'
NMR (400 MHz, dmso-d6): 1.31-1.51 ( l OH, m), 2.17-2.37 (6H, m),
2.98-3.09 (2H, m), 6.36 (1H, t), 7.44 (1H, d), 7.82 (1H, s).
1 -(2-Fluoro-4-Bromo phenyl)-3-(3 piperidin-1 yl propyl)-urea
Prepared via the general procedure for urea synthesis (via isocyanate)
Yield: 86%
C15H21BrFN3O Mass (calculated) 358; (found) [M+H+]= 358-360
Lc Rt= 2.18 (100%), 10'
NMR (400 MHz, dmso-d6): 1.31-1.40 (2H, m), 1.41-1.49 (4H, m),
1.50-1.57 (2H, m), 2.17-2.35 (6H, m), 3.07 (2H, q, J=5.8Hz), 6.59 (1H, t),
7.25 (1H, dt), 7.46 (1H, dd), 8.08 (1H, t), 8.34 (1H, s).
1- (2-Chloro-4-Bromo phenyl)-3-(3 piperidin-1 yl propyl)-urea
Prepared via the general procedure for urea synthesis (via isocyanate)
Yield: 78%
C15H21BrC1N3O Mass (calculated) 374; (found) [M+H+]= 374-376
Lc Rt= 2.46 (98%), 10'
NMR (400 MHz, dmso-d6): 1.30-1.39 (2H, m),1.42-1.49 (4H, m),
1.50-1.60 (2H, m), 2.16-2.37 (6H, m), 3.08 (2H, q, J=4.7), 7.01(1H, t), 7.40
(1H, dd), 7.62 (1 H, d), 8.48 (1 H, s), 8.11 (1 H, d).
General procedure for amide synthesis from co-haloalkanoyl chlorides
O I + Br Br
Br H2N Br H
Br
R1,N
H
R2
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One-pot with low molar excess of amine R1R2NH
In a round-bottom 2-neck flask, triethylamine (1 eq) was added to a
solution of aryl or heteroaryl amine (1 eq) in a volume of DCE such as to
obtain a 1.2 M solution of amine; 5-bromovaleryl chloride (0.95 eq) was then
added dropwise as a solution in 1.2 M solution in DCE and the reaction was
stirred at room temperature for 1 hour 30 minutes. A 1.8 M solution of amine
R 1 R2NH (3 eq) and triethylamine (1 eq) in DCE were then added and the
reaction mixture strirred at 55 C for a time between 4 and 16 h, until LCMS
monitoring showed reaction completion. After this period the reaction mixture
was partitioned between water and DCM; the organic layer was washed with
saturated NaCI and dried over Na2SO4. The crude amides obtained after
solvent evaporation at reduced pressure were purified by trituration from Et20
or by flash chromatography.
One-pot with high molar excess of amine RJR2NH
Alternatively, a solution of aniline (1 eq) and triethylamine (1 eq) in
dichloromethane (0.2 mmol/mL) was cooled at 0 C under nitrogen
atmosphere. 5-Bromo-pentanoyl chloride (1 eq) in dichloromethane
(0.3 mmol/mL) was slowly added. The mixture was stirred at room
temperature for 1.5 hours, after which the amine R1R2NH (5 eq) and
triethylamine (1 eq) were added at once and the reaction was stirred at room
temperature for further 40 hours. The organic solution was washed with brine,
dried and the solvent removed. The product were triturated by hexane/diethyl
ether 1/1 or purified by flash chromatography.
One-pot, high molar excess of amine R1R2NH - array method
To a solution of aniline (1 eq for each molar equivalent of amine
R1 R2NH used) and triethylamine (1 eq for each molar equivalent of amine
R1R2NH used) in dichloromethane (0.3 mmol/mL for each amine R1R2NH
used) 5-bromo-pentanoyl chloride (1 eq for each molar equivalent of amine
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R1R2NH used) was slowly added and the mixture stirred at room temperature
for 2 hours. The solution was then split in as many aliquotes as many amines
used in the array and each portion added to a vial containing an amine
RI R2NH (5 eq) and triethylamine (1 eq). The reactions were then shaken at
5 room temperature for 40 hours. The organic solutions were washed with brine,
collected, dried (Na2SO4) and the solvent evaporated. The products were
purified by preparative HPLC.
Two steps, using an equimolar amount of amine R1R2NH
5-Bromopentanoic acid-(4-bromophenyl)-amide
10 4-Bromo-aniline (6 g, 0.035 mol) and 0.035 mol of NEt3 (4.87 mL)
were dissolved in 120 mL of dichloromethane and cooled at 0 C.
To this solution, 0.038 mol of 5-bromovaleryl chloride (5.4 mL) were
slowly added and the resulting mixture was stirred for 1 h at 0 C.
When all the starting material was consumed (as monitored by LCMS)
15 the solution was washed with 50 mL of Na2CO3 0.4 M and the organic layer
was recovered by extraction and drying over Na2SO4. The solvent was
removed under reduced pressure giving 10 g of the title compound as a white
solid (yield 86%).
C11H13Br2NO Mass (calculated) [335]; (found) [M+H+]=335
20 Lc Rt = 2.64, 100% (5 min method)
NMR (400 MHz, CDC13) 1.70-2.00 (4H, m), 2.35-2.45 (2H, m),
3.38-3.48 (2H, m), 7.30-7.50 (4H, m).
1 eq of the thus prepared alkylating agent was dissolved in butanone
(5-10 mL/mmol substrate) and to this 1 eq of NaI and 1.1 eq of the amine
25 R1R2NH were added. The mixture was stirred at 70 C or 24 hours. The
mixture was cooled. When the products precipitated as salts, they were taken
into water, free-based by addition of NaOH 10% to pH = 10 and extracted
with dichloromethane. In the case where no product precipitation occurred, the
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solvent was removed under reduced pressure, the crude was taken into
dichloromethane and extracted after adjusting the pH to 10 with NaOH 10%.
If necessary, the products were further purified by flash chromatography
5-Azepan-1 yl pentanoic acid (4-bromo-3 fluoro phenyl)-amide
Following the general procedure for amide synthesis, 3-fluoro-4-
bromoaniline (66 mg, 0.35 mmol) and triethylamine (35 mg, 0.35 mmol) were
dissolved in DCE (0.5 mL) and 5-bromovaleryl chloride (66 mg, 0.33 mmol)
in DCE (0.5 mL) was added dropwise. After 1.5 hours, azepane (118 mg,
0.105 mmol) and triethylamine (35 mg, 0.35 mmol) in DCE (0.5 mL) were
added and the reaction mixture heated at 55 C for 4 hours. Work-up followed
by prep HPLC afforded the title compound (87 mg, 77%) as the formate salt.
C 17H24BrFN2O Mass (calculated) [371.30]; (found)
[M+H+]=371.33/373.35.
LC Rt=2.23, 100% (10 min method)
NMR (400 MHz, CDC13): 1.7 (4H, s); 1.88-1.84 (8H, m); 2.44 (2H, mt);
2.98 (2H, m); 3.15 (4H, bs); 7.27 (1 H, m); 7.4 (1 H, dd, J=8.8,7.6); 7.80 (1
H,
dd, J=10.8, 2.4); 8.63 (1H, HCOOH,s); 9.8 (1H, bs).
5-Azepan-1 yl pentanoic acid (4-bromo-3-trifluoromethyl phenyl)-
amide
Following the general procedure, 4-bromo-3-trifluoromethyl-aniline
(82 mg, 0.35 mmol) and triethylamine (35 mg, 0.35 mmol) were dissolved in
DCE (0.5 mL) and 5-bromovaleryl chloride (66 mg, 0.33 mmol) in DCE
(0.5 mL) was added dropwise. After 1 h 30 min, azepane (118 mg,
0.105 mmol) and triethylamine (35 mg, 0.35 mmol) in DCE (0.5 mL) were
added, and the reaction mixture heated at 55 C for 4 hours. Work-up followed
by prep HPLC afforded the title compound (29 mg, 20%) as its formate salt.
C 18H24BrF3N2O Mass (calculated) [421.30]; (found)
[M+H+]=421.29/423.29.
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LC Rt=2.52, 98% (10 min method)
5-[Methyl-(2 pyridin-2 yl-ethyl)-amino]pentanoic acid (3-bromo-
phenyl)-amide
Prepared according to the general procedure for amide synthesis and
purified by preparative HPLC to afford 106 mg (47%) of the title compound
as the formate salt.
C19H24N3OBr. HCO2H Mass (calculated) [390.33-46.03]; found
[M+H+]=390.23,
Lc Rt=1.73, 100%
NMR (400 MHz, CD3OD): 1.73-1.89 (4H, m); 2.46-2.50 (2H, m); 2.91
(3H, s); 3.19-3.27 (4H, m); 3.49-3.55 (2H, m); 7.19-7.24 (2H, m), 7.31 (1H, br
dd, J=5.2 Hz, 6.8 Hz); 7.3 8(1 H, ddd, J=6.8 Hz, 1.6 Hz, 2.0 Hz); 7.80 (1 H,
ddd,
J=8.0 Hz, 7.6 Hz, 1.6 Hz); 7.91 (1 H, s); 8.46 (1 H, s); 8.51 (1 H, br d,
J=4.8 Hz).
S-(Methyl pentyl-amino) pentanoic acid (4-bromo phenyl)-amide
Prepared according to the general procedure for amide synthesis and
purified by preparative HPLC to afford 123 mg (60%) of the title compound
as the formate salt.
C17H27N2Obr HCOZH Mass (calculated) [355.32-46.03]; found
[M+H+]=355.27,
Lc Rt=1.98, 100%
NMR (400 MHz, CD3OD): 0.95 (3H, t, J=6.8 Hz); 1.32-1.44 (4H, m);
1.67-1.81 (6H, m); 2.43-2.49 (2H, m); 2.81 (3H, s); 3.04-3.13 (4H, m);
7.42-7.45 (2H, m), 7.49-7.52 (2H, m),.8.51 (1H, s).
General procedure for cross-coupling reaction with boronic acids
0 R
~/ Br (OH)26 ~ p I~
R1. ~~ l~
N
R2 H Pd(Ph3)4 Rt.N"/IA~N /
R2 H ~ I
R
A=CHZ,N
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Thermal conditions on ureas (A = N)
To a degassed solution of aryl or heteroaryl bromide prepared following
the general procedure for urea synthesis described above (1 eq), the
appropriate boronic acid (1.3 eq) was added dissolved in 40 volumes (i. e.
1 mL/g substrate) of acetonitrile/0.4N aqueous Na2CO3 (1/1), Pd[(PPh3)]4
(10% mol). The solution was refluxed overnight under nitrogen either in a
round-bottom flask or in a glass test tube in a Buchi SynCore apparatus.
The acetonitrile phase was separated and the desired products purified
over a SCX or silica column. Fractions containing the desired product were
combined and dried under reduced pressure.
Microwave conditions on ureas (A = N)
To a degassed solution of bromide prepared following the general
procedure for urea synthesis (1 eq), the appropriate boronic acid (1 eq) and
Na2CO3 (3 eq) in 20 volumes (i.e. 1 mL/g substrate) of acetonitrile/water
(1/1), Pd[(PPh3)]4 (10% mol) were added.
The solution was irradiated in a microwave oven using the following
parameters: power: 200 watt; ramp time: 1 min; hold time: 20:00 min;
temperature: 90C; pressure: 200 psi. The acetonitrile phase was separated, the
solvent was removed under reduced pressure and the crude material purified
using SCX column (eluting with a gradient of DCM/MeOH, MeOH,
NH3/MeOH). The fractions containing product were combined and dried under
reduced pressure.
Microwave conditions on amides (A = CH2)
To a degassed mixture of 5-alkyl-pentanoic acid aryl-amide (0.1 g,
1 eq) aryl boronic acid (1.1 eq) in acetonitrile/sodium carbonate 0.4M
solution
1/1 (4 mL), a catalytic amount of Pd[(PPh3)]4 (5 mmol%) was added. The
reaction mixture was heated at 90 C for 20 minutes under microwave
irradiation (150 Watt, pressure max) and then again other 20 minutes. The
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organic layer was separated and concentrated, and further purified by SCX
column and/or by preparative HPLC. The solvent was removed under reduced
pressure to afford the corresponding product.
General procedure for cross-coupling reaction with aryl/heteroaryl
bromides
Br OII ~ C R
R1.Ni~"Al~N B~O~ ~ O I~
R2 H Pd(Ph3)a R1.N~~A~N /
42 H I
R
A= CHZ, N
To a degassed mixture of w-aminoalkanoic acid [4-(4,4,5,5-tetramethyl-
[1,3,2]dioxaborolan-2-yl)-phenyl]-amide (0.16 g, 0.4 mmol,l eq) (prepared
following the general procedures described above) and an aryl/heteroaryl
bromide (0.4 mmol, 1 eq) in acetonitrile/sodium carbonate 0.4M solution 1/1
(8 mL) a catalytic amount of Pd[(PPh3)]4 (5 mmol %) was added. The reaction
mixture was stirred at 90 C for 18 hours under N2 and monitored for
formation of the product by LCMS. The organic layer was separated and
filtered on a Celite pad and the solvent removed under reduced pressure. The
crude was purified by preparative HPLC or flash chromatography.
General method for the synthesis of 1-(4-aryl phenyl)-3-(4-aminoalkyl-
butyl)-ureas via reductive amination and cross-coupling
a) 1- (4-Bromo phenyl)-3-(4, 4-diethoxy-butyl)-urea
A solution of 4-aminobutyraldehyde diethyl acetal (0.88 mL, 5 mmol)
in dry DCM (10 mL) was added over 30' to a solution of
4-bromophenylisocyanate (1 g, 5 mmol, 1 eq) in dry DCM (25 mL) cooled to
0 C (ice/water bath) under N2; after 1 hour a thick white precipitate of the
urea formed. The solid was recovered by filtration and washing with Et20
(3 X 20 mL) to yield 1-(4-bromo-phenyl)-3-(4,4-diethoxy-butyl)-urea in
essentially quantitative yield (1.78 g).
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C15H23BrN2O3 Mass (calculated) [359.27]; (found)
[M+Na+]=3 81.11/3 83.25
LC Rt=3.54, 100% (10 min method)
NMR (400 MHz, dmso-d6): 1.08 (6H, t, J=5.85 Hz); 1.42 (2H, m);
5 1.50 (2H, m); 3.06 (2H, m); 3.41 (2H, q); 3.54 (2H, q); 4.45 (1H, t); 6.19
(1H,
t); 7.37 (4H, s); 8.54 (1H, s).
b) 1-(4-Bromo phenyl)-3-(4-aminoalkyl-butyl)-ureas
1-(4-Bromo-phenyl)-3-(4,4-diethoxy-butyl)-urea (0.72 g, 2 mmol, 1 eq)
was dissolved in dry DCM (10 mL) at room temperature and Montomorrilonite
10 K-5 (0.145 g) was added. The reaction was stirred at room temperature for 2
hours, when LC-MS showed complete conversion into the aldehyde.
The reaction mixture was filtered to remove all solids and the amine (6
mmol, 3 eq) was added, followed by NaBH(OAc)3 (4 mmol, 2 eq). The
reaction was stirred at room temperature for 24 hrs.
15 Upon reaction completion (as monitored by LC-MS), the solvent was
removed under reduced pressure and the resulting residue was purified by
SCX column, eluting with DCM:MeOH 1:1 and then 2M NH3 in MeOH.
c) 1-(4-aryl phenyl)-3-(4-aminoalkyl-butyl)-ureas
To a degassed mixture of 1-(4-bromo-phenyl)-3-(4-aminoalkyl-butyl)-
20 ureas (0.11 g, 1 eq) and a substituted benzeneboronic acid or
pinacolboronate
ester (1.5 eq) in acetonitrile/sodium carbonate 0.4M solution 1/1 (3 mL) a
catalytic amount of Pd[(PPh3)]4 (5 mmol %) was added. The reaction mixture
was heated at 90 C for 10 minutes under microwave irradiation (150 Watt)
and then again other 10 minutes, if needed. The organic layer was separated
25 and purified by SCX column or by prep HPLC (standard acidic conditions).
1 -Methyl-4 piperidin-1 yl-butylamine
a) N-Boc-5-methyl-2-pyrrolidinone
A solution of di-tert-butyl carbonate (12.1 g; 55.5 mmol) in acetonitrile
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(20 mL) was added dropwise to a solution of 5-methyl-2-pyrrolidinone (5.0 g;
50 mmol) and 4-dimethylamino-pyridine (0.31 g; 5 mol%) in acetonitrile
(50 mL) at RT. The mixture was stirred for 7 hrs. The solvent evaporated and
the obtained crude dissolved in ethyl acetate and washed with NaHCO3 sat.
solution. The organic layer was collected and dried (Na2SO4) and evaporation
of the solvent gave the clean product.
9.41 g; 90%
C10H17NO3 calculated 199; found 126/144
Lc Rt (5 min)= 1.73, 99%
NMR (400 MHz, dmso-d6): 1.22 (3H, d, J= 6.4 Hz); 1.43 (9H, s);
1.50-1.57 (1H, m); 2.04-2.14 (1H, m); 2.26 (1H, ddd, J= 3.2 Hz, 9.2 Hz, 17.2
Hz); 2.56 (1H, ddd, J= 9.2 Hz, 10.42 Hz, 17.2 Hz); 4.07-4.15 (1H, m).
b) (1-Methyl-4-oxo-4 piperidin-1-yl-butyl)-carbamic acid tert-butyl
ester
N-Boc-5-methyl-2-pyrrolidinone (0.7 g; 3.52 mmol) and piperidine
(1.55 g; 18.2 mmol, 1.8 mL) were mixed and heated at 150 C by MW
irradiation for 40 minutes. The mixture was diluted with dichloromethane and
the solution washed trice with HC1 1.0 M solution. The organic layer was
dried (Na2SO4) and the solvent evaporated. The crude product was purified by
flash chromatography (cyclohexane: ethyl acetate).
0.522 g; 73%
C15H28N203 calculated 284; found 285
Lc Rt (3 min)= 1.31, 99%
NMR (400 MHz, dmso-d6): 0.99 (3H, d, J= 6.8 Hz); 1.35 (9H, s); 1.36-
1.40 (2H, m); 1.43-1.48 (2H, m); 1.50-1.58 (4H, m); 2.15-2.30 (2H, m); 3.30-
3.46 (5H, m); 6.63 (1H, d, J= 8.8 Hz).
c) 4-Amino-1 piperidin-1 yl pentan-l-one
(1-Methyl-4-oxo-4-piperidin-1-yl-butyl)-carbamic acid tert-butyl ester
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(3.50 g; 12.3 mmol) was dissolved in CH2C12 (40 mL) at 0 C and HC1 37%
solution (6 mL) were slowly added. After 15 minutes the ice bath was
removed and the mixture stirred at RT for 4 hrs. NaOH 2N solution was
slowly added to reach pH>8 and the organic layer collected, dried and the
solvent evaporated.
2.0 g; 88%
C10HZON20 calculated 184; found 185
Lc Rt (5 min)= 0.35, 100%
NMR (400 MHz, dmso-d6): 0.95 (3H, d, J= 6.4 Hz); 1.32-1.57 (8H, m);
2.20-2.36 (2H, m); 2.68-2.76 (1H, m); 3.21 (4H, br m).
d) 1 -Methyl-4 piperidin-1 yl-butylamine
4-Amino-l-piperidin-l-yl-pentan-l-one (2.0 g, 10.9 mmol) was
dissolved in THF (50 mL) at 0 C and a solution of lithium aluminum hydride
1.0 M in THF (22 mL, 22 mmol) was slowly added. The mixture was stirred at
0 C for 30 min, then 4 hrs at RT. Water (0.8 mL) was added to the solution
and the solvent evaporated. The residue was treated with ether and NaOH 15%
solution. The organic layer was collected and dried. Evaporation of the
solvent
gave a product clean enough to be used without further purification.
1,21 g; 65%
CIoH22N2 calculated 170; found 171
Lc Rt (5 min)= 0.35, 100%
NMR (400 MHz, dmso-d6): 0.89 (3H, d, J= 6.4 Hz); 1.10-1.17 (2H, m);
1.30-1.37 (4H, m); 1.39-1.45 (6H, m); 2.10-2.14 (2H, m); 2.23 (4H, br m);
2.65-2.69 (1H, m). .
2, 2-Difluoro-4 piperidin-1-yl-butylamine
a) 3,3-Difluoro-succinamic acid
To a solution of 2,2-Difluoro-succinic acid (2.0 g, 13 mmol) in 20 mL
of iPrOAc, Trifluoroacetic anhydride (2.2 mL, 15.6 mmol) was added in one
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portion.
The solution was stirred under N2 atmosphere at 50 C for 1 hour. The
formation of the 2,2-Difluoro-succinic anhydride was confirmed by LCMS
after quenching of a little part with methanol [C5H6F204 calculated 168; found
M- 167; Lc Rt (5 min)= 1.05].
The solution was then cooled at room temperature and added dropwise
to NH3 in methanol (7N, 15 mL, 105 mmol) under N2 atmosphere maintaining
the temperature below 20 C.
The solvent was then evaporated and the residue dissolved in a small
amount of water basified with Na2CO3 (pH 8-9). The aqueous phase was
washed with EtOAc, acidified with HC1 6N to pH 1 and the product extracted
several time with Et20 and CHC13.
The organic layers were collected and dried. Evaporation of the solvent
gave 0.877 g of crude product used without further purification.
Yield: 45%
C4H5FZNO3 calculated 153; found M- 152
Lc Rt (5 min)= 0.32
NMR (400 MHz, dmso-d6): 3.16 (2H, t, J = 15.4); 7.86 (1H, s); 8.09
(1 H, s); 12.90 (1 H, s).
b) 2,2-Difluoro-4-oxo-4 piperidin-1 yl-butyramide
3,3-Difluoro-succinamic acid (0.18 g, 1.2 mmol) was dissolved in
10 mL of acetonitrile and the mixture was cooled at 0 C under N2 atmosphere.
N,N-dicyclohexylcarbodiimide (0.266 g, 1.3 mmol) was added and the
mixture was stirred again at 0 C for further 10 minutes.
1-Hydroxybenzotriazole hydrate (0.308 g, ca. 2 mmol) was then added and the
ice bath removed.
After 20 minutes at RT Piperidine (0.115 mL, 1.2 mmol) was added and
the reaction was stirred at RT overnight.
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The solvent was evaporated and the residue dissolved in
Dichloromethane. The organic phase was washed with HCl 0.16 N and then
with water.
The organic solvent was then evaporated and the residue purified by
SiO2 column (eluant: EtOAc:DCM 9:1).
0.113 g of pure product were obtained.
Yield: 43%
C9HI4F2N202 calculated 220; found M+ 221
Lc Rt (5 min)= 1.16
NMR (400 MHz, dmso-d6): 1.35-1.57 (6H, m); 3.25-3.36 (6H, m); 7.71
(1H, s); 7.92 (1H, s).
c) 2,2-Difluoro-4 piperidin-1-yl-butylamine
To solution of 2,2-Difluoro-4-oxo-4-piperidin-l-yl-butyramide
(0.323 g, 1.5 mmol) in 10 mL of dry THF was slowly added lithium aluminum
hydride solution 1.0 M in THF (5.88 mL, 5.9 mmol) under Nitrogen
atmosphere at 0 C. The mixture was stirred for 3 days at RT. The excess of
hydride was then quenched with water and the solvent removed. The residue
was then diluted with methanol and filtered on Celite The solution was
purified by SCX column and the obtained oil was used without further
purification.
0.169 g of product were obtained.
Yield: 59%
C9HI8F2N2 calculated 192; found M+ 193
Lc Rt (5 min)= 0.19
19F-HNMR (400 MHz, dmso-d6): -106.16 (quint.)
2,2-Dimethyl-4 piperidin-1 yl-butylamine
a) 2,2-Dimethyl-4-oxo-4 piperidin-1 yl-butyric acid
To solution of 2,2-dimethylsuccinic anhydride (1.0 g, 7.8 mmol) and
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triethylamine (0.79 g, 078 mmol) in CH2C12 (40 mL) was slowly added a
solution of piperidine (0.66 g, 7.8 mmol) in CH2C12 (10 mL) at RT. The
mixture was stirred for 4 hrs. NaOH 1.OM solution was added and the aqueous
layer collected and subsequently acidified to pH 4 using HC1 2.0 M solution.
5 Extraction with CHC13 and evaporation of the solvent gave a crude product
clean enough to be used without further purification.
1.63 g; 98%
C11H19NO3 calculated 213; found M+ 214/M- 212
Lc Rt (5 min)= 1.56
10 NMR (400 MHz, dmso-d6): 1.10 (6H, s); 1.32-1.37 (2H, m); 1.40-1.44
(2H, m); 1.50-1.55 (2H, m); 2.50 (2H, s); 3.31-3.35 (4H, m); 11.7 (1H, br s).
b) 2, 2-Dimethyl-4-oxo-4 piperidin-1 yl-butyramide
To a solution of 2,2-dimethyl-4-oxo-4-piperidin-1-yl-butyric acid
(2.0 g, 9.4 mmol) in CH2C12 (40 mL) was slowly added oxalyl chloride
15 (2.98 g, 23.5 mmol) under nitrogen atmosphere at RT. The mixture was
stirred for 3 hrs. After evaporation of the solvent and the excess of oxalyl
chloride, the obtained crude product was dissolved in CH2C12 (20 mL) and
added to a solution of ammonia 0.5 M in dioxane (200 mL). After addition of
NaHCO3 sat. solution, the organic layer was extracted twice, collected and
20 dried. Evaporation of the solvent gave a crude product purified by flash
chromatography (CH2C12/CH30H=96/4).
0.93 g; 46%
C 11H2ON202 calculated 212; found M+ 213
Lc Rt (5 min)= 1.32
25 NMR (400 MHz, CD3OD): 1.27 (6H, s); 1.48-1.54 (2H, m); 1.56-1.61
(2H, m); 1.63-1.69 (2H, m); 2.64 (2H, s); 3.45-3.49 (4H, m).
c) 2,2-Dimethyl-4 piperidin-1 yl-butylamine
To solution of 2,2-dimethyl-4-oxo-4-piperidin-1-yl-butyramide (2.0 g,
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9.43 mmol) in THF was slowly added lithium aluminum hydride solution
1.0 M in THF (28.3 mL) at 0 C under nitrogen atmosphere. The mixture was
stirred for 6 hrs. Water (1.0 mL) was added to the solution and the solvent
evaporated. The residue was treated with ether and NaOH 1.0 M solution, the
organic layers collected and dried. Evaporation of the solvent gave 1.4 g of
crude product used without further purification.
1.40 g; 80%
C> >H24N2 calculated 185; found M+ 186
Lc Rt (10 min)= 0.21
NMR (400 MHz, dmso-d6): 0.75 (6H, s); 1.26-1.29 (2H, m); 1.32-1.36
(2H, m); 1.41-1.47 (4H, m); 2.13-2.17 (2H, m); 2.25 (4H, br m); 2.26 (2H, br
s).
4-Piperidin-1 yl pentylamine
a) 2- (4-Bromo pentyl)-isoindole-1, 3-dione
1,4-Dibromo-pentane (2.9 g, 15,7 mmol) was added to a solution of
potassium phthalimide in 2-butanone at RT and the mixture stirred at RT 24
hrs, then 10 hrs at 50 C. After cooling the solvent was evaporated and the
crude purified by flash chromatography (cyclohexane/ethyl acetate).
4.0 g; 86%
NMR (400 MHz, Acetone): 1.68 (3H, d, J= 6.8 Hz); 1.80-1.93 (4H, m);
3.69 (2H, m); 4.26-4.34 (1 H, tq, J= 6.8 Hz, 4.8 Hz); 7.85 (4H, br m).
b) 2- (4-Piperidin-1 yl pentyl)-isoindole-1, 3-dione
A mixture of 2-(4-bromo-pentyl)-isoindole-1,3-dione (0.3 g, 1 mmol),
piperidine (0.1 mL, 1 mmol) and potassium carbonate (0.14 g, 1 mmol) in
DMF (3 mL) were mixed in a pressure tube at 80 C for 6 hrs. After filtration
the solvent was evaporated and the obtained residue purified by flash
chromatography (hexane/ethyl acetate).
0.19 g; 62%
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NMR (400 MHz, dmso-d6): 0.82 (3H, d, J= 6.4 Hz); 1.30-1.45 (8H, m);
1.53-1.62 (2H, m); 2.20-2.25 (2H, m); 2.35-2.40 (2H, m); 2.52-2.55 (1H, m);
3.54 (2H, m); 7.80-7.86 (4H, br m).
c) 4-Piperidin-1 yl pentylamine
2-(4-Piperidin-l-yl-pentyl)-isoindole-1,3-dione (0.19 g, 0.63 mmol) was
dissolved in ethanol. After addition of hydrazine hydrate (0.06 g, 1.27 mmol)
the mixture was stirred at reflux for 6 hrs and then left at RT overnight.
After
filtration of the solid, the organic solution was concentrated in vacuum and
the
obtained crude product purified by scx-column.
0.05 g; 46%
NMR (400 MHz, dmso-d6): 0.84 (3H, d, J= 6.4 Hz); 1.28-1.46 (10H,
m); 2.25-2.30 (2H, m); 2.37-2.51 (5H, m).
General 3-amino-5-aryl/heteroaryl pyrazole synthesis
The 3-amino-5-aryl/heteroaryl pirazoles used in the Examples were
eithere commercially available os synthesised using the routes shown in the
Scheme below:
0 0
Al or A1 bis
ArA 0 Ar
~ CN
N
NHZ
Ar
0 Bl CI B2
Ar/II`
CN
General procedure for aryl/heteroaryl fl-ketonitrile synthesis (A 1):
O CH3CN 0
ArA 0 NaH, toluene Ar
I CN
To a solution of an aryl or heteroaryl methyl carboxylate (6.5 mmol) in
dry toluene (6 mL) under N2, NaH (50-60% dispersion in mineral oil, 624 mg,
13 mmol) was carefully added. The mixture was heated at 80 C and then dry
CH3CN was added dropwise(1.6 mL, 30.8 mmol). The reaction was heated for
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18 hours and generally the product precipitated from the reaction mixture as
Na salt.
The reaction was then allowed to cool down to room temperature and
the solid formed was filtered and then dissolved in water. The solution was
then acidified with 2N HC1 solution and at pH between 2-6 (depending on the
ring substitution on the aryl/heteroaryl system) the product precipitated and
was filtered off. If no precipitation occurred, the product was extracted with
DCM.
After work-up, the products were generally used in the following step
without further purification. The general yield was between 40 and 80%.
General procedure for aryl/heteroaryl 8-ketonitrile synthesis (route
AI bis):
0
0 CH3CN or RCH2CN H, R
Ar
Ar 0 guLi, toluene CN
To a solution of dry alkanenitrile in toluene (1 mmol/mL, 5 eq.) cooled
down to -78 C under nitrogen, a solution of n-butyllithium in n-hexane (1.6 N,
3.5 eq) was added dropwise. The mixture was left stirring at -78 C for
minutes and then a solution of the aryl or heteroaryl methyl carboxylate in
toluene (0.75 mmol/mL, 1 eq.) was added and the reaction allowed to reach room
20 temperature. Upon reaction completion, after about 20 minutes, the mixture
was
cooled down to 0 C and HC1 2N was added to pH 2. The organic phase was
recovered, dried over Na2SO4 and concentrated under reduced pressure,
affording
the title product which was generally used without further purification.
General procedure for aryl aminopyrazole synthesis (route A2):
0 H
Ar-1Y H' R NI-N NH
1 2
CN Ar/ ~
H,R
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To a solution of the (3-ketonitrile (7.5 mmoL), in absolute EtOH
(15 mL) hydrazine monohydrate (0.44 mL, 9.0 mmol) was added and the
reaction was heated at reflux for 18 hrs. The reaction mixture was allowed to
cool to room temperature and the solvent was evaporated under reduced
pressure. The residue was dissolved in DCM and washed with water.
The organic phase was concentrated under reduced pressure to give a
crude product that was purified by Si02 column or by precipitation from Et20.
Yields were generally between 65 and 90%.
5- (IH-Indol-5 yl)-2H pyrazol-3 ylamine
a) 1-Triisopropylsilanyl-IH-indole-5-carboxylic acid methyl ester
To a solution of lg of methyl indole-5-carboxylate (5.7 mmol) in 10 mL
of dry DMF 273 mg of NaH (mineral oil dispersion 50-60%, 5.7 mmol) were
added and the mixture cooled to 0 C. Triisopropylchlorosilane (1.06 g,
5.7mmol) were added drop wise and after 1 hour LC-MS showed complete
conversion of the starting material to the title product. The mixture was
diluted
with 30 mL of DCM and washed with saturated Na2CO3. The organic phase
was dried over Na2SO4 and concentrated under reduced pressure. The crude was
purified with Si02 column eluting with n-hexane. The title compound was
obtained (500 mg, yield 26%).
C19H29NO2Si
Mass (calculated) [331]; (found) [M+H+]=332
LC Rt=3.39, 100% (5 min method)
'H-NMR: (dmso-d6): 1.06 (d, 18H, J=7.52), 1.75 (quin, 3H, J=7.52),
6.75 (m, 1H), 7.48 (m, 1H), 7.60 (m, 1H), 7.72 (m, 1H), 8.25 (s, 1H).
b) 3 -Oxo-3-(1-triisopropylsilanyl-IH-indol-5 yl) propionitrile
To a solution of 393 gL of anhydrous CH3CN (7.5 mmol) in 6 mL of
dry toluene cooled down to -78 C, 5.35 mLof butyllithium in hexane solution
(1.6 N) were added dropwise. The mixture was left stirring at -78 C for
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20 minutes and then a solution of 500 mg of 1-triisopropylsilanyl-lH-indole-
5-carboxylic acid methyl ester (1.5 mmol) in 2 mL of dry toluene were added
and the reaction allowed to reach room temperature. Upon reaction completion
after about 20 minutes the mixture was cooled down to 0 C and HC1 2N was
5 added to pH 2. The organic phase was separated, dried over Na2SO4 and
concentrated under reduced pressure, affording 490 mg of title product which
was used in the next step without further purification (yield = 96%).
C20H28N2OSi
Mass (calculated) [340]; (found) [M+H+]=341 [M-H+]=339
10 LC Rt=3.10, 89% (5 min method)
'H-NMR: (dmso-d6): 1.06 (18H,d, J=7.52), 1.76 (3H,quin, J=7.52), 4.76
(1H, d), 7.78-7.81 (1H, m), 7.48-7.52 (1H, m), 7.60-7.73 (2H, m), 8.25 (s,
1H).
c) 5- (IH-Indol-S yl)-2H pyrazol-3 ylamine
To a solution of 3-Oxo-3-(1-triisopropylsilanyl-lH-indol-5-yl)-
15 propionitrile (490 mg, 1.44 mmol) in 15 mL of absolute EtOH, 720 L of
hydrazine monohydrate (14.4 mmol) were added and the reaction refluxed for
18 hours. LC-MS showed complete conversion to the aminopyrazole and also
silyl deprotection. The mixture was concentrated under reduced pressure, and
purified with Si02 column (eluent gradient from 100% DCM to DCM:MeOH
20 9:1) to afford the title compounds (120mg, yield: 41 %).
CiIHioNa
Mass (calculated) [198]; (found) [M+H+]=199
LC Rt=0.84, 100% (3 min method)
5-Pyridin-3 yl-2H pyrazol-3 ylamine
25 a) 3-Oxo-3 pyridin-3 yl propionitrile
The product was prepared according to the general procedure for
aminopyrazole synthesis (route Al).
'H-NMR (400 MHz, MeOH-d4): 9.07 (1H, d), 8.81 (2H, dd), 8.26 (1 H,
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dt), 7.59 (1 H, dd), 4.79 (2H, s).
b) 5-Pyridin-3 yl-2H pyrazol-3 ylamine
The product was prepared according to general procedure for
aminopyrazole synthesis (route A2).
The crude product was purified with Si02 column (5 g) with gradient
elution from 100% DCM to DCM-NH3 (2N MeOH solution) 95:5. The title
product (371 mg, 68% yield) was obtained.
1H-NMR (400 MHz, MeOH-d4): 8.82 (1 H, d), 8.41 (1 H, dd), 7.98 (1 H,
dt), 7.3 7(1 H, dd), 5.82 (2H, s)
3-Imidazo[1,2-a]pyridin-6 yl-3-oxo propionitrile
The product was obtained starting from imidazo[1,2-a]pyridine-6-
carboxylic acid methyl ester according to general procedure Al
Yield 39%
C10H7N3O Mass (calculated) [185]; (found) [M+H+]=186 [M-H]=184
LC Rt=0.23, 100% (3 min method)
1H-NMR: (dmso-d6): 4.72 (2H,s), 7.61-7.65 (2H, m), 7.70 (1H, m),
8.07 (1H, s), 9.40 (s, 1H).
5-Imidazo[1,2-a]pyridin-6 yl-lH-pyrazol-3 ylamine
The title compound was synthesized according to general procedure A2
starting 3-imidazo[1,2-a]pyridin-6-yl-3-oxo-propionitrile
Yield: 84%
C l OH9N5 Mass (calculated) [ 199]; (found) [M+1 ]= 200
LCMS, (5min method, RT=0.21 min,
NMR (1H, 400MHz, MeOH-d4) 3,34 (s, 2H), 5,90 (br s, 1H), 7,57
(s, 1H), 7,63 (br s, 1H), 7,86 (s, 1H), 8,73 (s, 1H).
5- (3-Fluoro phenyl)-IH pyrazol-3 yl-amine
a) 3- (3-Fluoro phenyl)-3-oxo proprionitrile
The product was prepared according to a modification of general route
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Al. To a solution of methyl-3-fluorobenzoate (3 g, 18 mmol) in dry toluene
(25 mL) under N2, NaH (50-60% dispersion in mineral oil, 1.44 g, 36 mmol)
was carefully added.
The mixture was heated at 90 C and then dry CH3CN was added
dropwise (4.45 mL, 85.2 mmol). The reaction was heated for 18 hours and the
product precipitated from the reaction mixture as its sodium salt. The
reaction
was allowed to cool down to room temperature and the solid formed was
filtered, then redissolved in water, and the solution was acidified with 2N
HC1
to pH 5-6, upon which precipitation was observed. Filtration of the solid from
the aqueous solution afforded 2.12 g of the title compound (72% yield) which
was used directly in the following step.
b) 5- (3-Fluoro phenyl)-1H pyrazol-3 yl-amine
The product was prepared according to a slight modification of route
A2. To a solution of 3-(3-fluoro-phenyl)-3-oxo-propionitrile (1.92 g,
11.77 mmoL) in absolute EtOH (32 mL) hydrazine monohydrate (0.685 mL,
14.12 mmol) was added and the reaction was heated at reflux for 2 hrs. The
reaction mixture was allowed to cool to room temperature and the solvent was
evaporated under reduced pressure. The crude was treated with ether and
filtered to give 1.71 g of title compound were recovered (82% yield).
C9H8FN3
Mass (calculated) [177]; (found) [M+H+] =190
LC Rt = 1.13, 69% (5 min method)
5-Pyridin-4-yl-1 H pyrazol-3 ylamine
3- Oxo-3 pyridin-4-yl propionitrile
The product was prepared according to a modification of route Al. To a
solution of 3 g (22 mmol) of isonicotinic acid methyl ester in dry toluene
(30 mL) under N2, NaH (50-60% dispersion in mineral oil, 1.75 g, 44 mmol)
was carefully added.
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The mixture was heated at 90 C and then dry CH3CN was added
dropwise (5.39 mL, 103 mmol). The reaction was heated for 18 hours and the
product precipitated from the reaction mixture as the sodium salt. The
reaction
was allowed to cool down to room temperature and the solid formed was
filtered, then it was dissolved in water and the solution was acidified with
6N
HC1 solution to pH 5-6 and the product extracted with DCM. The pH of the
aqueous phase was adjusted again to 4-5 and another extraction with DCM
afforded more product.
The organic phases were combined, dried and evaporated. The product
was used directly in the following step. Yield of crude product: 58%.
b) 5-Pyridin-4 yl-IH pyrazol-3 ylamine
The product was prepared according to a modification of route A2. To a
solution of 3-oxo-3-pyridin-4-yl-propionitrile (1.86 g, 12.74 mmoL) in
absolute EtOH (35 mL) hydrazine monohydrate (0.74 mL, 15.29 mmol) was
added and the reaction was heated at reflux for 2 hours. The reaction mixture
was then allowed to cool to room temperature and the solvent was evaporated
under reduced pressure. The crude product obtained was washed with ether to
afford the title compound (yield: 39%).
C8H8N4
Mass (calculated) [160]; (found) [M+H+] =161
LC Rt = 0.23, 100% (5 min method)
'H-NMR (400 MHz, dmso-d6): 5.02 (2H, s); 5.85 (1H, s); 7.59 (2H, d,
J=6 Hz); 8.50 (2H, d, J=6 Hz); 11.93 (1H, s).
Chlorocynnamonitrile synthesis (route B1)
o ci
Arlk Ar
CN
POC13 (2 eq with respect to the aryl/heteroaryl acetophenone) were added
dropwise to 4 molar equivalents of anhydrous DMF cooled down to 0 C, at such
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a rate that the temperature did not exceed 10 C. The acetophenone (1 eq) was
then added dropwise and the reaction was allowed to reach room temperature.
The reaction was then stirred for further 30' and then 0.4 mmol of
hydroxylamine hydrochloride were added. The reaction was then heated up to
50 C, after which heating was removed and additional 4 eq. of hydroxylamine
hydrochloride were added portionwise (at such a rate that the temperature
never exceeded 120 C). The reaction was then stirred until the temperature of
the mixture spontaneously decreased to 25 C. Water (100 mL) were then
added and the mixture was extracted with diethyl ether. The organic phase was
dried over Na2SO4 and concentrated under reduced pressure. The crude
product was used for the next step without further purification.
Aryl aminopyrazole synthesis (route B2)
ci
HN-N
Ar k NHz
CN Ar
To a solution of the chlorocynnamonitrile (0.5 mmol/mL, 1 eq) in
absolute EtOH 2 eq of hydrazine monohydrate were added and the reaction
was heated at reflux for 4 hrs. The reaction mixture was allowed to cool to
room temperature and the solvent was evaporated under reduced pressure. The
residue was triturated with Et20, allowing to recover the title compound which
was generally used without further purification.
5- (3-Bromo phenyl)-2H pyrazol-3 ylamine
a) 3-(3-Bromo-phenyl)-3-chloro-acrylonitrile
To 30.9 mL of dry DMF (400 mmol) cooled down to 0 C 18.3 mL of
POC13 (200 mmol) were added dropwise so that the temperature was always
under 10 C. To the mixture 19.9 g (100 mmol) of 1-(3-bromophenyl)ethanone
were added dropwise and the reaction was allowed to reach room temperature.
When the addition was complete the reaction was stirred for further
minutes and then 2.7 g (40 mmol) of hydroxylamine hydrochloride were
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added and the reaction heated up to 50 C. The heating was then removed and
other 27 g (400 mmol) of hydroxylamine hydrochloride were added
portionwise (so that the temperature did never exceed120 C).
After the last addition the reaction was left stirring until the temperature
5 of the mixture spontaneously decreased to 25 C. Water (100 mL) was then
added and the mixture was extracted with diethyl ether. The organic phase was
dried over Na2SO4 andconcentrated under reduced pressure.
The crude product was used for the next step without further
purification.
10 C9H5BrC1N
'H-NMR (400 MHz, dmso-d6): 7.03 (s, 1H), 7.44-7.54 m, 1H),
7.72-7.84 (m, 2H), 8.00 (br s, 1 H).
Yield 68%
b) 5- (3-Bromo phenyl)-2H pyrazol-3 ylamine
15 To a solution of 3-(3-bromo-phenyl)-3-chloro-acrylonitrile (10 mmoL),
in absolute EtOH (20 mL) hydrazine monohydrate (1 mL, 20 mmol) was
added and the reaction was heated at reflux for 4 hrs. The reaction mixture
was then allowed to cool to room temperature and the solvent was evaporated
under reduced pressure. The residue was triturated with Et20, allowing to
20 recover 1.8 g of the title compound as pure product (yield 54%).
C9H8BrN3
'H-NMR(400 MHz, dmso-d6): 4.58, 5.03 (1H, 2 tautomeric peaks),5.64,
5.84 (1H, 2 tautomeric peaks), 7.28 (1H, s), 7.35 (1H, s), 7.53-7.65 (1H, m),
7.77 (1H, s), 11.56, 11.97 (1H, 2 tautomeric peaks).
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General method for the synthesis of co-bromo-alkanoic acid
(IH pyrazol-3 yl-S-aryl)-amides
HzN I \ R
N-N
H
~O( ~ DIPEA Br 0
CI' Mn Br DMA, -10 C R
2hrs nH <\
N-NH
n=1,2,3
A solution of co-bromoalkanoyl chloride (15.7 mmol, 1 eq) in dry DMA
(35 mL) was cooled to -10 C (ice/water bath) under N2; a solution of
5-aryl/heteroaryl-lH-pyrazol-3-ylamine (15.7 mmol, 1 eq) and
diisopropylethylamine (15.7 mmol, 1 eq) in dry DMA (15 mL) is added over
30'. After 2 hrs at -10 C, completion of the reaction as monitored by LC-MS
was generally observed (acylation on the pyrazole ring is also detected). The
reaction is then quenched by addition of H20 (ca. 50 mL); the thick white
precipitate formed upon addition of water was recovered by filtration.
Washing with Et20 (3 X 10 mL) usually efficiently removed the byproduct of
acylation on the pyrazole ring.
General method for the synthesis of w-amino-alkanoic acid
(IH pyrazol-3 yl-5-aryl)-amides
R1
P-H
Br O R2 R1
~
~R DIPEA, Nal 2 O
R
R
N_N DMF, +50 C \ N \ y
n=1 2 3 18 hrs N-N
u)-Bromo-alkanoic acid [5-aryl-1H-pyrazol-3-yl]-amide (0.6 mmol,
1 eq) is dissolved in DMF (4 mL), sodium iodide (0.6 mmol, 1.0 eq) is added
followed by the secondary amine (1.5 mmol, 2.5 eq) and
diisopropylethylamine (0.6 mmol, 1 eq). The reaction is then stirred under N2
at + 50 C for 18 hrs.
Upon reaction completion (as monitored by LC-MS), the solvent is removed
at reduced pressure and the resulting oily residue is dissolved in DCM (20
mL),
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washed with sat. Na2CO3 (2 X 20 mL) and sat. NaC1 (2 X 20 mL); the organic
layer is dried over Na2SO4 and the solvent removed under reduced pressure. The
title compounds were purified either by silica column or preparative HPLC.
General synthetic method for the one-pot synthesis of aw-amino-alkanoic
acid (IH pyrazol-3-yl-5-aryl)-amides: acylation-nucleophilic substitution
N
1 ~ R
0 "N R
R2, R1 R1 OII
~ f~~ - Br DIPEA N
CI" ~L Jn Br N-~\ --~ RpiN~ fi~ J~ N ~ R
DMA, -10 C N'N 60-C or rt
2 hrs N-N
n=1,2,3
To a solution of co -bromoalkanoyl chloride (0.94 mmol, leq) in DMA
(1mL) cooled at 0 C is added a solution of 3-amino-5-aryl/heteroarylpyrazole
(0.94 mmol, leq) and diisopropylethylamine (1.88 mmol, 2 eq) in DMA (2 mL)
and the reaction is stirred for 1 hour at 0 C. The secondary amine (2.35 mmol,
2.5 eq) and NaI (0.94mmol, 1 eq) are then added. For 3-carbon chain
derivatives the reaction was generally complete after 2 hours at room
temperature. For 4-carbon chain derivatives the reaction mixture was generally
heated at 60 C for 24-48 hours. Upon complete conversion of the bromo-
intermediate (as monitored by LC-MS), the solvent was removed under reduced
pressure. The residue was taken up in DCM (2 mL) and washed with Na2CO3
saturated water solution. The organic phase was concentrated under reduced
pressure and the crude products were either recrystallised from CH3CN, or
purified by Si02 column (gradient from 100%DCM to DCM-NH3MeOH 2N
solution 8:2) or by preparative HPLC (standard acidic conditions).
General method for the synthesis of to-amino-alkanoic acid
(lhl pyrazol-3 yl-5-aryl)-amides via the amino acid route
HZN R
0 X 0 NaOH ^ OH N-N H
Br,,OnJ~OEt X1-11 Qn-,k OEt X Qn~ -~' X1--, On~N ~\ R
HCI CDI 0 N-N
H
n-1'2'3
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To a solution of amine X (65 mmol) in toluene (15 mL) ethyl co -
bromoalkanoate (26 mmol) was added and the reaction mixture was refluxed
for 10 hours. The mixture was allowed to cool to room temperature and any
solid present was filtered off and washed with ether. The filtrate was
concentrated under reduced pressure to give the co-aminoester which was used
in the next step without further purification.
To a suspension of crude ethyl co-aminoalkanoate from the previous
step (about 25 mmol) in 15 mL of water, NaOH (1.4 g, 25 mmol) was added
and the mixture was heated at reflux for 16 hours. The reaction was then
allowed to cool down to room temperature, the solution was acidified at 0 C
with HC1 6N and concentrated under reduced pressure. The residue was
treated with EtOH and the sodium chloride which precipitated was filtered off.
Evaporation of the solvent under reduced pressure afforded the co-aminoacid
as a white solid. To a suspension of co-aminoacid (7.93 mmol) in 12,2-
dichloroethane (20 mL), N,N'-carbonyldiimidazole (1.2 g, 7.4 mmol) was
added and the mixture was stirred at room temperature for 2 hours (when all
the aminoacid was activated complete dissolution of the suspension was
generally observed). The 3 -amino- 5 -aryl/heteroarylpyrazole (5.29 mmol) was
then added and the reaction was stirred for further 10 hours. Upon reaction
completion (as monitored by LC-MS) if the formation of two isomers was
observed, the mixture was heated at 50 C until the conversion of the less
stable isomer to the title compound was observed (as monitored by LC-MS).
The solvent was washed with sat. Na2CO3 solution, extracted and removed
under reduced pressure. The crude products were either recrystallised from
CH3CN, or purified by Si02 column or by preparative HPLC.
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Example 1
5-Azepan-1 yl pentanoic acid (2-chloro-2 ; 3'-difluoro-biphenyl-4 yl)-
amide
a) 5-Azepan-1-y1 pentanoic acid (4-bromo-3-chloro-phenyl)-amide
Following the general procedure, 4-bromo-3-chloroaniline (72 mg,
0.35 mmol) and triethylamine (35 mg, 0.35 mmol) were dissolved in DCE
(0.5 mL) and 5-bromovaleryl chloride (66 mg, 0.33 mmol) in DCE (0.5 mL)
was added dropwise. After lh 30 min, azepane (118 mg, 0.105 mmol) and
triethylamine (35 mg, 0.35 mmol) in DCE (0.5 mL) was added and the
reaction mixture heated at +55 C for 4 hours. Work-up followed by
preparative HPLC afforded the title compound (23 mg, 17%) as formate.
C17H24BrC1N2O Mass (calculated) [387.75]; (found) [M+H+]=389.28.
LC Rt=2.34, 100% (10 min method)
b) 5-Azepan-1 yl pentanoic acid (2-chloro-2 ; 3'-difluoro-biphenyl-4 yl)-
amide
Following the general procedure for cross-coupling under microwave
conditions, 5-azepan-1-yl-pentanoic acid (4-bromo-3-chloro-phenyl)-amide
(70 mg, 0.18 mmol) and 2,3-difluorophenyl boronic acid (31 mg, 0.2 mmol)
were dissolved in acetonitrile/0.4 M Na2CO3 1/1 (1.5 mL each) and
Pd[P(Ph)3]4 (11 mg, 0.01 mmol) was added. After irradiating for 20 minutes,
preparative HPLC purification afforded the title compound (23 mg, 27%) as
the corresponding formate, as a white solid.
C23H27C1F2N20 Mass (calculated) [420.93]; (found) [M+H+]=421.38
LC Rt= 2.04, 100% (10 min method)
NMR (400 MHz, CDC13): 1.63 (4H, s); 1.80 (8H, m); 2.43 (2H, m);
2.94 (2H, m); 3.1 (4H, bs); 6.96 (1H, m); 7.05-7.02 (2H, m); 7.11 (1H, d,
J=8); 7.16 (1 H, d, J=8.4); 7.5 3(1 H, d, J=8.4); 7.9 (1 H, s); 8.56(1 H, s,
HCOOH); 9.68 (1H, s, NH).
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Example 2
4'-{S-[Methyl-(2 pyridin-2 yl-ethyl)-amino]pentanoylamino}-biphenyl-
3-carboxylic acid amide
Prepared according to the general procedure for amide synthesis
5 followed by the general procedure for cross-coupling under microwave
irradiation to give 0.07 g of title compound (62%).
C26H30N402. Mass (calculated) [430.55]; (found) [M+H+]=431.42
Lc Rt=1.59, 100%
NMR (400 MHz, dmso-d6): 1.37-1.45 (2H, m); 1.51-1.59 (2H, m); 2.17
10 (3H, s); 2.28-2.36 (4H, m); 2.62-2.66 (2H, m), 2.81-2.85 (2H, m); 7.14 (1H,
br
dd, J=7.6 Hz, 4.8 Hz); 7.24 (1 H, d, J=8.0 Hz); 7.40 (1 H, br s); 7.48 (1 H,
dd,
J=8 Hz, 7.6 Hz); 7.61-7.7.0 (5H, m); 7.78 (2H, m); 8.06 (1 H, s); 8.11 (1 H,
s);
9.96 (1H, s).
Example 3
15 1-(2, 2 '-Dimethoxy-biphenyl-4-yl)-3-(4 piperidin-1 yl-butyl)-urea
1 -(4-Bromo-3 -methoxy-phenyl)-3 -(4-piperidin-1-yl-butyl)-urea
(prepared according to the general procedure for urea synthesis, reaction with
isocyanate) was weighed into a microwave vessel (100 mg, 0.26 mmol) and
dissolved in acetonitrile (1 mL). To this, 2-methoxyphenylboronic acid (47
20 mg, 0.312 mmol) was added, along with
tetrakis(triphenylphosphine)palladium (20 mg, 0.017 mmol) and a solution of
sodium carbonate (1 mL, 0.4 M). The reaction mix was then exposed to
microwave irradiation at psi 250, at 90 C for 20 minutes. On reaction
completion by LCMS, the separated organic phase was removed from the
25 reaction mix and passed through a plug of Celite . The collected crude was
loaded onto an SCX column, eluting the desired compound with a solution of
ammonia in methanol (20% ammonia). Fractions containing the desired
compound were combined and dried affording the titled compound (30 mg,
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0.073 mmol, 28% yield).
C24H33N303 Mass (calculated) [411.55]; (found) [M+H+] =412
LC Rt=2.03, 98% (10 min method)
NMR (400 MHz, CDC13): 1.57-1.67 (11H, m); 2.42-2.50 (5H, m); 3.29
(2H, m); 3.77 (6H, s); 5.85 (1H, s); 6.61 (1H, s); 6.75-6.77 (1H, d); 6.95-
7.00
(2H, m); 7.15 (1 H, d); 7.21-7.22 (1 H, m) 7.23-7.33 (2H, m)
Example 4
5-Imidazol-1 yl pentanoic acid (3'-acetylamino-biphenyl-4 yl)-amide
a) 5-Bromopentanoic acid- (4-bromophenyl) -amide
4-Bromo-aniline (6 g, 0.035 mol) and 0.035 mol of NEt3 (4.87 mL)
were dissolved in 120 mL of dichloromethane and cooled at 0 C.
To this solution, 0.038 mol of 5-bromovaleryl chloride (5.4 mL) were
slowly added and the resulting mixture was stirred for 1 h at 0 C.
When all the starting material was consumed (monitoring by LCMS) the
solution was washed with 50 mL of Na2CO3 0.4 M and the organic layer was
recovered by extraction and drying over Na2SO4. The solvent was removed
under reduced pressure giving 10 g of the title compound as a white solid
(yield 86%).
C11H13Br2NO Mass (calculated) [335]; (found) [M+H+]=335
Lc Rt = 2.64, 100% (5 min method)
NMR (400 MHz, CDC13) 1.70-2.00 (4H, m), 2.35-2.45 (2H, m),
3.38-3.48 (2H, m), 7.30-7.50 (4H, m).
b) 5-Im idazolyl acid- (4-bromophenyl) -am ide
400 mg (1.19 mmol) of 5-bromopentanoic acid-(4-bromophenyl)-amide
and 800 mg of imidazole (11.7 mmol) were suspended in 2 mL of a mixture of
Toluene: EtOH 1:1. The reaction mixture was heated at 160 C for 10 minutes
under microwave conditions. The solvent was removed under reduced pressure
and the crude mixture dissolved in DCM (10 mL) and washed twice with 1M
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NaOH. The solution was dried over Na2SO4, filtered and the solvent removed
to give the desired product as a white powder (240 mg yield 62%).
NMR (400 MHz, dmso-d6): 7.65 (1H, s), 7.56-7.52 (2H, m), 7.46-7.42
(2H, m), 7.42 (1H, m), 6.87 (1H, m), 3.95 (2H, t, J = 12 Hz), 2.30 (2H, t, J
7.2 Hz), 1.75-1.68 (2H, m), 1.55-1.45 (2H, m).
c) 5-Imidazol-1-yl pentanoic acid (3'-acetylamino-biphenyl-4 yl)-amide
A mixture of 1 mL of 0.4 M solution of Na2CO3 and 1 mL of DMF was
added to a microwave tube containing 5-imidazolyl acid-(4-bromophenyl)-
amide (100 mg, 0.31 mmol), 3-acetoamidophenyl boronic acid (83.4 mg, 0.46
mmol) and [Pd(PPh3)4] (16 mg, 0.03 mmol). The reaction mixture was heated
at 90 C for 20 minutes under microwave conditions. The mixture was diluted
with MeOH and the solution passed through a plug of Celite . The solvent
was removed under reduced pressure and the crude mixture dissolved in DCM
and washed with 1M NaOH. The organic phase was dried over MgSO4,
filtered and the solvent removed under reduced pressure. Purification by SCX
and after by crystallization from EtOAC/Et20 gave 43 mg of pure product
(yield 37%).
C22H24N402 Mass (calculated) [376]; (found) [M+H+]= 377
Lc Rt 2.11 (10 min)
Purity 98%
NMR (400 MHz, dmso-d6): 10.03 (1 H, s), 8.85 (1 H, d, J = 2.0 Hz),
8.50 (1 H, dd, J= 4.8 Hz, 1.6 Hz), 8.03 (1 H, m), 7.67 (4H, m), 7.63 (1 H, s),
7.44 (1 H, m), 6.87 (1H, m), 3.98 (2H, t, J = 7.2 Hz), 2.34 (2H, t, J= 7.2
Hz),
1.74 (2H, m), 1.53 (2H, m).
Example 5
1- (2,2'-Difluoro-biphenyl-4-yl)-3-(4 piperidin-1 yl-butyl)-urea
a) 1- (4-Bromo-3 fluoro phenyl)-3-(4 piperidin-1 yl-butyl)-urea
Prepared via the general procedure for urea synthesis (triphosgene
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activation of aniline).
Yield: 71%
C 16H23BrFN3O Mass (calculated) 372; (found) [M+H+]= 372-374
Lc Rt= 2.26 (100%), 10'
NMR (400 MHz, dmso-d6): 128-1.56 (IOH, m), 2.16-2.38 (6H, m), 3.00-
3.15 (2H, m), 6.30 (1 H, t), 6.99 (1 H, dd), 7.46 (1 H, t), 7.59 (1 H, dd),
8.81 (1 H,
s).
b) 1- (2, 2'-Difluoro-biphenyl-4 yl)-3-(4 piperidin-1 yl-butyl)-urea
To a degassed solution of 1-(4-bromo-3-fluoro-phenyl)-3-(4-piperidin-
1-yl-butyl)-urea (100 mg, 0.26 mmol) in DME/H20 (1.8 mL/0.3 mL)
2-fluorophenyl boronic acid (55 mg, 0.39 mmol), Na2CO3 (55 mg, 0.52
mmol), Pd(OAc)2 (6 mg, 10% mol) and tri-o-tolylphosphine (34 mg, 20%
mol), were added. The solution was irradiated under microwave conditions for
minutes with power 200W.
15 The organic phase was diluted with 1 mL of AcOEt and separated
The crude was purified with prep-HPLC (36 mg, 40% yield).
Molecular formula: C22H27F2N30
Mass (calculated) [387]; (found) [M+H+]=388
Lc Rt (10 min method)= 399, 97%
20 'H-NMR (400MHz, d6-DMSO): 1.27-1.54 (IOH, m), 2.25-2.44 (6H, m),
2.94-3.13 (2H, m), 6.33-6.45 (1H, m), 7.09-7.14 (1H, m), 7.21-7.30 (3H, m),
7.34-7.45 (2H, m), 7.48-7.55 (1H, m), 8.16 (1H, s), 8.85 (1H, s).
Example 6
3'-Fluoro-4'-[3-(4 piperidin-1 yl-butyl)-ureido]-biphenyl-3-carboxylic
acid amide)
a) 1- (2-Fluoro-4-bromo phenyl)-3-(4 piperidin-1 yl-butyl)-urea
Prepared via the general procedure for urea synthesis (isocyanate)
Yield:88%
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NMR (400 MHz, dmso-d6): 1.22-1.50 (lOH, m), 2.12-2.37 (6H, m),
3.00-3 .13 (2H, m), 6.62 (1 H, t), 7.25 (1 H, d), 7.47 (1 H, dd), 8.10 (1 H,
t), 8.33
(1H, s)
b) 3 '-Fluoro-4'-[3-(4 piperidin-1 yl-butyl)-ureido]-biphenyl-3-
carboxylic acid amide
To a degassed solution of 1-(2-fluoro-4-bromo-phenyl)-3-(3-piperidin-
1-yl-propyl)-urea (100 mg, 0.27 mmol), the 3-benzamide-phenylboronic acid
(66 mg,0.44 mmol) and Na2CO3 (3 eq) in 20 volumes (weight/vol) of
acetonitrile/water (1/1), Pd[(PPh3)]4 (10% mol) were added.
The solution was irradiated in a microwave oven using the following
parameters:
power: 200 watt; ramp time: 1 min; hold time: 20:00 min; temperature:
90 C; pressure: 200 psi.
The acetonitrile phase was separated, the solvent was removed under
reduced pressure and the crude material purified using SCX column (eluting
with a gradient of DCM/MeOH, MeOH, NH3/MeOH). The fractions
containing the desired product were combined and dried under reduced
pressure, and then further purified by preparative HPLC (yield 15%).
Molecular formula: C22H27F2N30
Mass (calculated) [387]; (found) [M+H+]=388
Lc Rt (10 min method)= 399, 97%
'H-NMR (400MHz, CD3OD): 1.40-1.67 (IOH, m), 2.26-2.58 (6H, m),
3.21-3.27 (2H, m), 7.13-7.20 (1H, m), 7.21-7.28 (1H, m), 7.30-7.39 (1H, m),
7.40-7.51 (2H, m), 7.56 (1H, s,), 8.11-8.16 (1H, m).
Example 7
4'-{5-[Methyl-(2 pyridin-2 yl-ethyl)-amino]pentanoylamino}-biphenyl-
3-carboxylic acid amide
Prepared according to the general method for amide coupling (one-pot,
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excess amine followed by the general method for cross-coupling with boronic
acids, microwave conditions) to give 0.07 g (yield = 62%) of title compound.
C26H30N402. Mass (calculated) [430.55]; (found) [M+H+J=431.42
Lc Rt=1.59, 100%
5 NMR (400 MHz, dmso-d6): 1.37-1.45 (2H, m); 1.51-1.59 (2H, m); 2.17
(3H, s); 2.28-2.36 (4H, m); 2.62-2.66 (2H, m), 2.81-2.85 (2H, m); 7.14 (1H, br
dd, J=7.6 Hz, 4.8 Hz); 7.24 (1 H, d, J=8.0 Hz); 7.40 (1 H, br s); 7.48 (1 H,
dd,
J=8 Hz, 7.6 Hz); 7.61-7.7.0 (5H, m); 7.78 (2H, m); 8.06 (1H, s); 8.11 (1H, s);
9.96 (1 H, s).
10 Example 8
1-(4'-Methoxy-biphenyl-4 yl)-3-(1-methyl-4 piperidin-1 yl-butyl)-urea
a) 1- (4-Bromo phenyl)-3-(1-methyl-4 piperidin-1 yl-butyl)-urea
To a solution of 1-methyl-4-piperidin-1-yl-butylamine (1.07 g, 6.3
mmol) in CHZC12 (15 mL) at 0 C was slowly added a solution of 4-
15 bromophenyl isocyanate (1.25 g, 6.3 mmol) in CHZC12 (15 mL). The mixture
was stirred 30 min at OC, then 3 hrs at RT. The solvent was evaporated and the
residue purified by SCX column. The obtained solid was washed with ether.
1.57 g; 68%
C17H26BrN3O calculated 368; found 368/370
20 Lc Rt (5 min)= 1.35, 100%
NMR (400 MHz, dmso-d6): 1.02 (3H, d, J= 6.8 Hz); 1.30-1.45 (5H, m);
2.13-2.17 (5H, m); 2.13-2.17 (2H, m); 2.23 (4H, br m); 3.57-3.64 (1H, m);
5.97 (1H, J= 8 Hz); 7.29-7.34 (4H, m); 8.36 (1H, s).
b) 1-(4'-Methoxy-biphenyl-4-yl)-3-(1-methyl-4 piperidin-1 yl-butyl)-
25 urea
To a degassed mixture of 1-(4-bromo-phenyl)-3-(1-methyl-4-piperidin-
1-yl-butyl)-urea (0.3 g, 0.81 mmol) and 4-methoxyphenylboronic acid
(186 mg, 1.22 mmol) in acetonitrile/sodium carbonate 0.4M solution 1/1
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(8 mL) a catalytic amount of Pd[(PPh3)]4 (47 mg, 5 mmol %) was added. The
reaction mixture was heated at 90 C for 20 minutes under microwave
condition. After addition of ethyl acetate (1 mL), the organic layer was
separated and purified by scx columns.
0.27 g; 84%
C17H26BrN3O calculated 395; found 396
Lc Rt (10 min)= 2.23, 100%
NMR (400 MHz, dmso-d6): 1.05 (3H, d, J= 6.4 Hz); 1.33-1.47 (IOH,
m); 2.06-2.18 (2H, m); 2.26 (4H, br m); 3.61-3.67 (1H, m); 3.75 (3H, s); 5.95
(1H, J= 8 Hz); 6.95-6.97 (2H, m); 7.39-7.46 (4H, m); 7.50-7.52 (2H, m); 8.30
(1H, s).
Example 9
1-(2, 2-Dimethyl-4 piperidin-1 yl-butyl)-3-[4-(1-methyl-1 H pyrazol-4-
yl) phenylJ-urea
a) 1- (4-Bromo phenyl)-3-(2,2-dimethyl-4 piperidin-1 yl-butyl)-urea
To a solution of 2,2-dimethyl-4-piperidin-1-yl-butylamine (1.4 g,
7.6 mmol) in CH2C12 (20 mL) at 0 C was slowly added a solution of
4-bromophenyl isocyanate (1.5 g, 7.6 mmol) in CH2C12 (15 mL). The mixture
was stirred 20 min at 0 C, then 4 hrs at RT. The solvent was evaporated and
the residue treated with ether and the solid separated. The solution was
purified by SCX column. The obtained solid was used without further
purification.
0.825 g (28%)
C18H28BrN3O calculated 382; found M+ 382/384
Lc Rt (5 min)= 1.39
NMR (400 MHz, dmso-d6): 0.80 (6H, s); 1.28-1.34 (4H, m); 1.41-1.46
(4H, m); 2.17-2.21 (2H, m); 2.28 (4H, br m); 2.90 (2H, d, J= 6Hz); 6.15 (1H,
t, J= 6 Hz); 7.3 3-7.3 5(4H, m); 8.51 (1 H, s).
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b) 1-(2, 2-Dimethyl-4 piperidin-1 yl-butyl)-3-[4-(1-methyl-1 H pyrazol-
4 yl) phenylJ-urea
To a degassed mixture of 1-(4-bromo-phenyl)-3-(2,2-dimethyl-4-
piperidin-1-yl-butyl)-urea (0.11 g, 0.29 mmol) and 1-methyl-4-(4,4,5,5-
tetramethy11,3,2-dioxaborolan-4-yl)-1 H-pyrazole (90 mg, 0.43 mmol) in
acetonitrile/sodium carbonate 0.4M solution 1/1 (4 mL) a catalytic amount of
Pd[(PPh3)]4 (17 mg, 5 mmol %) was added. The reaction mixture was heated
at 90 C for 20 minutes under microwave condition. After addition of ethyl
acetate (1 mL), the organic layer was separated and purified by scx columns
followed by preparative HPLC.
0.067 g; 61%
C22H33N50 calculated 383; found M+ 384
Lc Rt (10 min)= 1.61
NMR (400 MHz, dmso-d6): 0.82 (6H, s); 1.35-1.40 (4H, m); 1.48-1.54
(4H, m); 2.40-2.44 (2H, m); 2.46-2.50 (4H, br m); 2.91 (2H, d, J= 6Hz); 3.81
(3H, s); 6.32 (1H, t, J= 6 Hz); 7.33-7.39 (4H, m); 7.72 (1H, s); 7.97 (1H, s);
8.23 (1H, s); 8.56 (1H, s).
Example 10
1 -(4-Piperidin-1 yl pentyl)-3-(4 pyridin-3 yl phenyl)-urea
a) 1 -(4-Bromo phenyl)-3-(4 piperidin-1 yl pentyl)-urea
To a solution of 4-piperidin-1-yl-pentylamine (0.20 g, 1.18 mmol) in
CH2C12 (4 mL) at 0 C was slowly added a solution of 4-bromophenyl
isocyanate (0.23 g, 1.18 mmol) in CH2C12 (2 mL). The mixture was stirred
5 min at 0 C, then 1 hrs at RT. The solvent was evaporated and the residue
used without further purification.
0.39 g; 90%
NMR (400 MHz, CD3OD): 0.96 (3H, d, J= 6.4 Hz); 1.25-1.59 (10H, m);
2.46-2.55 (5H, m); 3.11 (2H, t, J= 6.8 Hz); 7.25-7.28 (2H, m); 7.32-7.36 (2H,
m).
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b) 1- (4-Piperidin-1 yl pentyl)-3-(4 pyridin-3 yl phenyl)-urea
To a degassed mixture of 1-(4-bromo-phenyl)-3-(4-piperidin-1-yl-
pentyl)-urea (130 mg, 0.35 mmol) and pyridine-3-boronic acid (65 mg, 0.53
mmol) in acetonitrile/sodium carbonate 0.4M solution 1/1 (4 mL) a catalytic
amount of Pd[(PPh3)]4 (17 mg, 5 mmol %) was added. The reaction mixture
was heated at 90 C for 10 minutes under microwave condition. After addition
of ethyl acetate (1 mL), the organic layer was separated and purified by scx
columns followed by preparative HPLC.
0.025 g; 19%
C22H30N40 calculated 366; found 367
Lc Rt (10 min)= 0.24-0.81
NMR (400 MHz, dmso-d6): 0.82 (6H, d, J= 6.8); 1.24-1.55 (10H, m);
2.46-2.68 (5H, m); 3.05-3.09 (2H, m); 6.38 (1H, t, J= 5.6 Hz); 7.41 (1H, ddd,
J= 8.0 Hz, 4.8 Hz, 0.8 Hz); 7.49-7.52 (2H, m); 7.57-7.60 (2H, m); 7.99 (1 H,
ddd, J= 8.0 Hz, 2.0 Hz, 1.6 Hz); 8.21 (1 H, s); 8.47 (1 H, dd, J= 4.8 Hz, 1.6
Hz); 8.74 (1H, s); 8.82 (1 H, dd, J= 2.0 Hz, 0.8 Hz).
Example 11
1-[4-(1-Methyl-1 H pyrazol-4-yl) phenylJ-3-[4-(3, 3, 3-trifluoro-
propylam ino)-butyl]-urea
a) 1-(4-Bromo phenyl)-3-[4-(3, 3, 3-trifluoro propylamino)-butyl]-urea
1-(4--Bromo-phenyl)-3-(4,4-diethoxy-butyl)-urea (0.72 g, 2 mmol, 1
eq) was dissolved in dry DCM (10 mL) at room temperature and
Montomorrilonite K-5 (0.145 g) is added. The reaction was stirred ar room
temperature for 2 hours, when LC-MS shows complete conversion into the
aldehyde. The reaction mixture was filtered to remove all solids and
trifluopropylamine.HCl (0.9 g, 6 mmol, 3 eq) and diisoproylethylamine
(1.05 mL, 6 mmol, 3 eq) were added, followed by NaBH(OAc)3 (1.2 g,
4 mmol, 2 eq,). The reaction was stirred at rt for 24 hrs. Upon reaction
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completion (as monitored by LC-MS), the solvent was removed under reduced
pressure and the resulting residue was purified by SCX column, eluting with
DCM:MeOH 1:1 and then 2M NH3 in MeOH. 1-(4-Bromo-phenyl)-3-[4-
(3,3,3-trifluoropropylamino)-butyl]-urea was obtained (0.33 g, 40% yield).
C14H19BrF3N3O Mass (calculated) [382.23]; (found) [M+H+]=381.25
(ESI-)
LC Rt=0.63, 90% (10 min method)
b) 1-[4-(1-Methyl-]M pyrazol-4 yl) phenylJ-3-[4-(3, 3, 3-trifluoro-
propylamino)-butyl]-urea
To a degassed solution of 1-(4-Bromo-phenyl)-3-[4-(3,3,3-
trifluoropropylamino)-butyl]-urea (0.11 g, 0.3 mmol), 31-Methyl-4-(4,4,5,5-
tetramethyl-[1,3,2]dioxaborolan-2-yl)-1H-pyrazole (0Ø94 g, 0.45 mmol) in
3 mL Acetonitrile/ 0.4 M Na2CO3 (50/50) Pd[(PPh3)]4 (0.02 mmol) were
added. The solution was irradiated under the microwave conditions described
above. The acetonitrile layer was separated and filtered on a Celite pad. The
solution was dried and the product purified by preparative HPLC to yield
mg of the title compound as formate salt (0.048 mmol, 16% yield).
C18H24F3N50 Mass (calculated) [383.42]; (found) [M+H+]= 384.21
Lc Rt= 1.48 (100%)
20 NMR (400 MHz, dmso-d6): 1.31 (4H, m); 2.43-2.30 (2H, m); 2.57-2.55
(2H, m); 2.75 (2H, m); 3.05 (2H, m); 3.81 (3H, s); 6.17 (1H, bs); 7.38-7.32
(4H, m); 7.72 (1H, s); 7.97 (1 H, s); 8.18 (1 H, s, HCOOH); 8.43 (1 H, s).
Example 12
4'-(S-Azepan-1 yl pentanoylamino)-2'-chloro-biphenyl-3-carboxylic
acid amide
a) 5-Azepan-1-yl pentanoic acid (4-bromo-3-chloro phenyl)-amide
4-bromo-3-chloroaniline (0.72 g, 3.5 mmol), triethylamine (0.48 mL,
3.35 mmol) and 5-bromovaleryl chloride (0.44 mL, 3.32 mmol) are reacted for
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2 hrs at rt in DCE (12 mL): after this time, azepane (1.18 mL, 10.3 mmol) and
more triethylamine (0.48 mL, 3.35 mmol) are added and the reaction stirred
for +55 C for 4 h. After reaction completion and work-up, the title compound
is clean enough (92% purity) for the following step (1.3 g, quant yield).
5 C17H24BrC1N2O Mass (calculated) [387.75]; (found) [M+H+]=
387.32/389.28
Lc Rt= 2.83 (92%), 10'
b) 4'-(5-Azepan-1-yl pentanoylamino)-2'-chloro-biphenyl-3-carboxylic
acid amide
10 5-Azepan-l-yl-pentanoic acid (4-bromo-3-chloro-phenyl)-amide
(0.07 g, 0.18 mmol) and benzamide-3-boronic acid (0.044 g, 0.27 mmol) are
reacted in acetonitrile/sodium carbonate 0.4M solution 1/1 (3 mL) with a
catalytic amount of Pd[(PPh3)]4 (5 mmol %).
The title compound is obtained as free base after purification by SCX
15 column (0.045 g, 0.11 mmol, 60% yield)
C24H30C1N302 Mass (calculated) [472.98]; (found) [M+H+]= 428.46
Lc Rt= 2.04 (100%), 10'
NMR (400 MHz, CDC13): 1.68-1.55 (8H, m); 1.77 (4H, m); 2.41 (2H,
m;, 2.58 (2H, m); 2.67 (2H, m); 2.72 (2H, m); 7.27 (1H, s); 7.49 (2H, m); 7.59
20 (1 H, s); 7.8 (2H, m); 7.85 (1 H, s); 8.16 (1 H, bs).
Example 13
5-Azepan-1 yl pentanoic acid (2 fluoro-2'-methoxy-biphenyl-4 yl)-
amide
a) 5 -Azepan-1 yl pentanoic acid (4-bromo-3 fluoro phenyl)-amide
25 4-Bromo-3-fluoroaniline (0.66 g, 3.5 mmol), triethylamine (0.48 mL,
3.35 mmol) and 5-bromovaleryl chloride (0.44 mL, 3.32 mmol) are reacted for
2 hrs at rt in DCE (12 mL): after this time, azepane (1.18 mL, 10.3 mmol) and
more triethylamine (0.48 mL, 3.35 mmol) are added and the reaction stirred
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for +55 C for 4 h. After reaction completion and work-up, the title compound
is clean enough (90% purity) for the following step (1.29 g, quant yield).
C 17H24BrFN2O Mass (calculated) [371.30]; (found) [M+H+]=
371.33/373.35
Lc Rt= 2.23 (90%), 10'
b) 5-Azepan-1 yl pentanoic acid (2 fluoro-2'-methoxy-biphenyl-4 yl)-
amide
5 -Azepan-1-yl-pentanoic acid (4-bromo-3 -fluoro-phenyl)-amide
(0.093 g, 0.25 mmol) and 2-methoxyphenyl-boronic acid (0.057 g, 0.37 mmol)
are reacted in acetonitrile/sodium carbonate 0.4M solution 1/1 (4 mL) with a
catalytic amount of Pd[(PPh3)]4 (5 mmol %).
The title compound is obtained as formate salt after purification by
preparative HPLC (0.025 g, 0.06 mmol, 13% yield).
C24H31FN202 Mass (calculated) [398.53]; (found) [M+H+]= 399.18
Lc Rt= 2.98 (98%), 10'
NMR (400 MHz, CD3OD): 1.83-1.72 (8H, m); 1.9 (4H, m); 2.5 (2H, m);
3.18 (2H, m); 3.37 (4H, m); 3.77 (3H, s); 6.99 (1H, m); 7.06 (1H, bs); 7.21
(1 H, m); 7.25 (2H, m); 7.3 5(1 H, m); 7.5 7(1 H, m); 8.49 (1 H, s, HCOOH).
Example 14
1-(2, 2-Difluoro-4 piperidin-1 yl-butyl)-3-(4'-methoxy-biphenyl-4 yl)-
urea
a) 1 -(4-Bromo phenyl)-3-(2, 2-difluoro-4 piperidin-1-yl-butyl)-urea
To a solution of 2,2-difluoro-4-piperidin-1-yl-butylamine (0.149 g,
0.78 mmol) in CH2C12 (4 mL) at 0 C a solution of 4-bromophenyl isocyanate
(0.153 g, 0.78 mmol) in CH2C12 (1 mL) was slowly added. The mixture was
stirred at 0 C for 30 min, then at room temperature until the reactants were
all
consumed. The solution was purified by SCX column. The oil obtained was
used without further purification (0.218 g).
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Yield: 72%
C 16H22BrF2N3O calculated 390; found M+ 390/392
Lc Rt (5 min)= 1.34
1H-NMR (400 MHz, dmso-d6): 1.43-1.57 (8H, m); 2.02-2.53 (8H, m);
7.22-7.43 (6H, m).
b) 1-(2,2-Difluoro-4 piperidin-1 yl-butyl)-3-(4'-methoxy-biphenyl-4 yl)-
urea
To a degassed mixture of 1-(4-bromo-phenyl)-3-(2,2-difluoro-4-
piperidin-1-yl-butyl)-urea (0.109 g, 0.28 mmol), 4-methoxyphenylboronic
(0.051 g, 0.34 mmol) and Na2CO3 (0.042 g, 0.40 mmol) in DME (1.3 mL)
and water (0.2 mL) tri-o-tolylphosphine (0.017 g, 20 mmol%) and Pd(OAc)2
(0.003 g, 5% mmol) were added. The reaction mixture was heated at 90 C for
10 minutes under microwave conditions. After addition of ethyl acetate
(1 mL), the organic layer was separated and purified by scx columns followed
by Si02 column (gradient from 100%DCM to DCM-NH3 in MeOH 2N
solution 9:1) to give 0.010 g of product.
Yield: 9%
C23H29F2N302 calculated 417; found M+ 418/M- 416
Lc Rt (10 min)= 2.23, 100%
NMR (400 MHz, dmso-d6): 1.47-1.62 (6H, m); 2.09-2.22 (2H, m);
2.46-2.62 (6H, m); 3.64 (2H, t, J=14); 3.81 (3H, s); 6.96 (2H, d, J=8.8);
7.40 (2H, d, J=6.8), 7.47-7.51 (4H, m).
19F-HNMR (400 MHz, dmso-d6): -118.90 (quint.)
Example 15
5-Piperidin-1 yl pentanoic acid [5-(4-methoxy phenyl)-4-methyl-2H-
pyrazol-3-yl]-amide
a) 3- (4-Methoxy phenyl)-2-methyl-3-oxo propionitrile
The product was prepared according to the general procedure for
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aminopyrazole synthesis (route Al).
The crude product was purified with Si02 column (10 g) with gradient
elution from 100% Hexane to Hexane-AcOEt 7:3. to give 1.43 g of pure
product (yield 31 %).
'H-NMR (400 MHz, MeOH-d4): 7.97 (2H, d), 6.98 (1H, d), 4.31 (1H, q,
J = 7.3 Hz), 3.89 (3H, s), 1.63 (3H, d, J = 7.3 Hz).
b) 5- (4-Methoxy phenyl)-4-methyl-2H pyrazol-3-ylamine
The product was prepared according to the general procedure for
aminopyrazole synthesis (route A2)
The crude product was purified with Si02 column (10 g) with gradient
elution from 100% DCM to DCM-MeOH 8:2. 1.0 g of pure product were
obtained (yield 65%).
1H-NMR (400 MHz, CDC13): 7.37 (2H, d), 6.97 (2H, d), 3.84 (3H, s),
2.03 (3H, s).
c) 5-Piperidin-1 yl pentanoic acid [5-(4-methoxy phenyl)-4-methyl-2H-
pyrazol-3 yl]-amide
The product was prepared according to the general synthetic method for
the one-pot synthesis of uo-amino-alkanoic acid (1H-pyrazol-3-yl-5-aryl)-
amides.
The crude product was purified with Si02 column (2 g) with gradient
elution from 100% DCM to DCM-NH3 (2N MeOH solution) 95:5.
The obtained crude was then purified again by prep-HPLC to give
54 mg of pure product (yield 7%).
C21H30N402
Mass (calculated) [370]; (found) [M+H+] =371
LC Rt=1.61, 100% (10 min method)
'H-NMR (400 MHz, dmso-d6): 9.57 (1 H, s), 8.12 (1H, s), 7.47 (2H, d),
7.02 (2H, d), 3.78 (3H, s), 2.41 (4H, broad), 2.37 (2H, m), 2.29 (2H, t), 1.91
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(3H, s), 1.57 (2H, m), 1.50 (6H, m), 1.38 (2H, m).
Example 16
N-[S-(4-Methoxy phenyl)-2H pyrazol-3 yl]-4 piperidin-1 yl-butyramide
a) 4-Piperidin-1-yl-butyric acid ethyl ester
To a solution of piperidine (5.4 g, 65 mmol) in toluene (15 mL) ethyl 4-
bromobutyrate (3.8 mL, 26 mmol) was added and the reaction mixture was
refluxed for 10 hours. The mixture was allowed to cool down to room
temperature and the white solid present (piperidium bromide) was filtered off
and washed with ether. The filtrate was concentrated under reduced pressure
to give the title product which was used in the next step without further
purification.
CIIH21NO2
Mass (calculated) [199]; (found) [M+H+] =200
LC Rt = 0.2, 100% (5 min method)
'H-NMR (400 MHz, MeOH-d4): 1.22-1.25 (3H, m), 1.46-1.47 (2H, m),
1.57-1.63 (4H, m), 1.78-1.84 (2H, m), 2.30-2.35 (4H, m), 2.42 (4H, m, broad),
4.08-4.14 (2H, m).
b) 4-Piperidin-1-yl-butyric acid
To a suspension of crude 4-piperidin-1-yl-butyric acid ethyl ester from
the previous step (about 25 mmol) in 15 mL of water, NaOH (1.4 g, 25 mmol)
was added and the mixture was heated at reflux for 16 hours. The reaction was
then allowed to cool down to room temperature, the solution was acidified at
0 C with HC1 6N and concentrated under reduced pressure. The residue was
treated with EtOH and the sodium chloride which precipitated was filtered off.
Evaporation of the solvent under reduced pressure afforded 2.8 g of the title
compound as a white solid in 58% overall yield of steps a) and b).
C9H NO2
Mass (calculated) [171]; (found) [M+H+] =172
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LC Rt = 0.23, 100% (5 min method)
'H-NMR (400 MHz, dmso-d6): 1.44-1.51 (2H, m); 1.64-1.80 (6H, m);
2.22-2.25 (2H, m); 2.75-2.78 (2H, m, broad); 2.91-2.94 (2H, m, broad);
3.30-3.40 (2H, m).
5 c) N-[S-(4-Methoxy phenyl)-2H pyrazol-3 yl]-4 piperidin-1 yl-butyramide
To a suspension of 4-piperidin-1-yl-butyric acid (1.32 g, 7.93 mmol) in
12,2-dichloroethane (20 mL), N,N'-carbonyldiimidazole (1.2 g, 7.4 mmol) was
added and the mixture was stirred at room temperature for 2 hours (when all
the aminoacid was activated complete dissolution of the suspension was
10 generally observed). 3 -Amino- 5 -(4-methoxyphenyl)pyrazole (1 g, 5.29
mmol)
was then added and the reaction was stirred for further 10 hours. Upon
reaction completion (as monitored by LC-MS) the formation of two isomers
was observed, and the mixture was heated at 50 C until the conversion of the
less stable isomer to the title compound was observed (as monitored by
15 LC-MS). The solvent was washed with sat. Na2CO3 solution, extracted and
removed under reduced pressure. The crude was crystallised from acetonitrile
to give 1.2 g of the title compound (Yield: 70%).
C i 9H26Na02
Mass (calculated) [342]; (found) [M+H+] =343
20 LC Rt = 1.54, 100% (10 min method)
'H-NMR (400 MHz, dmso-d6): 1.34-1.40 (1H, m); 1.52-1.55 (1H, m);
1.62-1.75 (6H, m); 1.94-1.98 (2H, m); 2.37-2.40 (2H, m); 2.81-2.88 (2H, m);
2.97-3.03 (2H, m); 3.39-3.42 (2H, m); 3.77 (3H, s); 6.77 (1H, s); 6.98 (2H, d,
J= 8.8 Hz); 7.61 (2H, d, J= 8.8 Hz); 10.47 (1 H, s), 12.66 (1 H, s).
25 Example 17
N-[S-(3-Methoxy phenyl)-]H-pyrazol-3 y1J-4-morpholin-4 yl-butyramide
a) 3- (3-Methoxy phenyl)-3-oxo propionitrile
To a solution of commercially available 3-methoxy-benzoic acid ethyl
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ester (3.2 g, 18 mmol) in dry toluene (25 mL), under N2, NaH (50-60%
dispersion in mineral oil, 1.44 g, 36 mmol) was carefully added. The mixture
was heated at 90 C and anhydrous CH3CN was added dropwise (4.45 mL,
85.2 mmol). The reaction was heated for 18 hours and the product precipitated
from the reaction mixture as Na salt. The reaction was allowed to cool down
to room temperature and the solid formed was filtered and washed with ether,
then it was redissolved in water and the solution acidified with 2N HC1
solution to pH 3 when precipitation of title compound was observed. Filtration
of the solid from the aqueous solution afforded 1.57 g of title product
(50% yield).
C10H9NOZ
Mass (calculated) [175]; (found) [M+H+] =176
LC Rt = 1.69, 94% (5 min method)
b) 5- (3-Methoxy phenyl)-2H pyrazol-3 ylamine
To a solution of 3-(3-methoxy-phenyl)-3-oxo-propionitrile
(8.96 mmoL) in absolute EtOH (20 mL) hydrazine monohydrate (0.52 mL,
15 mmol) was added and the reaction was heated at reflux for 18 hrs. The
reaction mixture was then allowed to cool to room temperature and the solvent
was evaporated under reduced pressure.
The crude was treated with ether and filtered, to give 1.4 g of title
product (83% of yield).
C1oH11N30
Mass (calculated) [189]; (found) [M+H+] =190
LC Rt = 1.13, 100% (5 min method)
'H-NMR (400 MHz, MeOH-d4): 3.82 (3H, s); 5.93 (1H, s); 6.86-6.88
(1 H, m); 7.19-7.31 (3H, m).
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c) N-[5-(3-Methoxy-phenyl)-1H pyrazol-3-y1]-4-morpholin-4 yl-
butyramide
A solution of 4-bromobutyryl chloride (0.104 mL, 0.9 mmol) in dry
DMA (1 mL) was cooled to -10 C (ice/water bath) under N2; 5-(3-methoxy-
phenyl)-2H-pyrazol-3-ylamine (170 mg, 0.9 mmol) and diisopropylethylamine
(0.315 mL, 1.8 mmol) in dry DMA (1 ml) were added. Upon complete
conversion to the intermediate 4-Bromo-N-[5-(3-methoxy-phenyl)-1H-
pyrazol-3-yl]-butyramide (as monitored by LC-MS), morpholine (0.079 mL,
0.9 mmol) was added and the mixture was heated at 60 C for 16 hours. The
residue was dissolved in DCM (2 mL) and washed with sat. Na2CO3 solution.
The organic phase was concentrated under reduced pressure and the crude
product was purified by Si02 column (gradient from Acetonitrile 100% to
MeCN/MeOH, NH3 90/10). The fractions containing the title compound were
collected to afford 17 mg (5.5% of yield).
C 18H24N403
Mass (calculated) [344]; (found) [M+H+] =345
LC Rt = 1.36, 95% (10 min method)
'H-NMR (400 MHz, MeOH-d4): 1.77-1.85 (2H, m); 2.34-2.40 (8H, m);
3.59-3.62 (4H, m); 3.76 (3H, s); 6.79-6.85 (2H, m); 7.15-7.29 (3H, m).
Example 18
4-Azepan-1 yl-N-[5-(3-methoxy phenyl)-]H pyrazol-3 ylJ-butyramide
A solution of 4-bromobutyryl chloride (0.104 mL, 0.9 mmol) in dry
DMA (1 mL) was cooled to -10 C (ice/water bath) under N2; 5-(3-Methoxy-
phenyl)-2H-pyrazol-3-ylamine (170 mg, 0.9 mmol) and diisopropylethylamine
(0.315 mL, 1.8 mmol) in dry DMA (1 ml) was added. Upon complete
conversion to the o)-bromoamide intermediate (as monitored by LC-MS)
0.101 mL of azepine were added to the solution and the mixture was left
stirring at 60 C for 16 hours.
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The residue was dissolved in DCM (2 mL) and washed with saturated
Na2CO3 solution. The organic phase was concentrated under reduced
pressure and the crude product was purified by Si02 column (gradient from
acetonitrile 100% to MeCN/MeOH, NH3 90/10). The fractions containing the
title product were collected and a further purification by preparative HPLC
was carried out to afford 20 mg of the title compound as its formate salt
(5.5% yield).
C20H28N402
Mass (calculated) [356]; (found) [M+H+] =357
LC Rt=1.71, 99% (10 min method)
'H-NMR (400 MHz, MeOH-d4): 1.65-1.68 (4H, m); 1.80-1.90 (4H, m);
1.97-2.04 (2H, m); 2.49-2.52 (2H, m); 3.12-3.16 (2H, m); 3.24-3.30 (4H, m,
broad); 3.75 (3H, s); 6.76 (1H, s); 6.82-6.85 (1H, m); 6.13-6.15 (2H, m);
6.23-6.27 (1H, m); 8.37 (1H, s, formate)
Example 19
4-Azepan-1-yl-N-[S-(4 fluoro phenyl)-2H pyrazol-3 ylJ-butyramide
Prepared - following the general synthetic method for the one-pot
synthesis of co-amino-alkanoic acid (1H-pyrazol-3-yl-5-aryl)-amides. Starting
from commercially available 5-(4-fluoro-phenyl)-2H-pyrazol-3-ylamine and
following the procedure, 25 mg of title compound were recovered as its
formate salt after preparative HPLC purification (7% yield).
C20H28N402
Mass (calculated) [344]; (found) [M+H+] =345
LC Rt=1.69, 100% (10 min method).
'H-NMR (400 MHz, MeOH-d4): 1.66-1.69 (4H, m); 1.80-1.90 (4H, m,
broad); 1.97-2.05 (2H, m); 2.52-2.54 (2H, m); 3.12-3.18 (2H, m); 3.25-3.30
(4H, m, broad); 6.67 (1H, s, broad); 7.08-7.12 (2H, m); 7.59-7.63 (2H, m);
8.43 (1 H, s, formate).
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Example 20
N-[S-(6-Methyl pyridin-3 yl)-]H-pyrazol-3 ylJ-4 piperidin-1 yl-
butyramide
a) 3- (6-Methyl pyridin-3 yl)-3-oxo propionitrile
The oxopropionitrile was synthesised following the general method for
3-oxopropionitriles (route Al).
CgH8N20
Mass (calculated) [160]; (found) [M+H+] =161
LC Rt = 0.63, 100% (5 min method)
'H-NMR (400 MHz, dmso-d6): 2.55 (3H, s); 4.65 (2H, s); 7.43-7.45 (m,
1); 8.13-8.16 (1H, m); 8.94-8.95 (1H, m).
b) 5- (6-Methyl pyridin-3 yl)-1H pyrazol-3 ylamine
The aminopyrazole was synthesised following the general method
described in route A2.
C9H l ON4
Mass (calculated) [174]; (found) [M+H+] =175
LC Rt = 0.23, 100% (5 min method)
c) N-[S-(6-Methyl pyridin-3 yl)-IH pyrazol-3 yl]-4 piperidin-1 yl-
butyramide
Prepared following the general synthetic method for the one-pot
synthesis of w-amino-alkanoic acid (1H-pyrazol-3-yl-5-aryl)-amides to afford
19 mg (6% yield) of title compound as its formate salt after preparative HPLC
purification.
C I 8H2sN50
Mass (calculated) [327]; (found) [M+H+] =328
LC Rt = 0.33, 100% (10 min method)
'H-NMR (400 MHz, MeOH-d4): 1.40-1.90 (6H, m); 2.30-2.54 (5H, m);
3.05-3.09 (4H, m); 3.20-3.24 (2H, m); 6.72 (1 H, s, broad); 7.3 0(1 H, d J
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8.0 Hz); 7.92-7.94 (1 H, m); 8.3 5(1 H, s, formate); 8.67 (1 H, s).
Example 21
N-[S-(S-Methyl pyridin-3-yl)-]H pyrazol-3-ylJ-4 piperidin-1 yl-
butyramide
5 a) 3- (S-Methyl pyridin-3 yl)-3-oxo propionitrile
The oxopropionitrile was synthesised following the general method for
3-oxopropionitriles (route Al).
C9HgN20
Mass (calculated) [160]; (found) [M+H+] =161
10 LC Rt = 0.63, 100% (5 min method)
1H-NMR (400 MHz, MeOH-d4): 2.55 (3H, s); 4.65 (2H, s); 7.43-7.45
(m, 1); 8.13-8.16 (1H, m); 8.94-8.95 (1H, m).
b) 5-(5-Methyl pyridin-3 yl)-1H pyrazol-3 ylamine
The aminopyrazole was synthesised following the general method
15 described in route A2.
C9H 10N4
Mass (calculated) [174]; (found) [M+H+] =175
LC Rt = 0.23, 100% (5 min method)
c) N-[5-(5-Methyl pyridin-3 yl)-]H pyrazol-3 ylJ-4 piperidin-1 yl-
20 butyramide
Prepared following the general synthetic method for the one-pot
synthesis of co-amino-alkanoic acid (1H-pyrazol-3-yl-5-aryl)-amides to afford
25 mg of the title compound as its formate salt (7.4% yield) after preparative
HPLC purification.
25 C18H25N50
Mass (calculated) [327]; (found) [M+H+] =328
LC Rt = 0.33, 100% (10 min method)
'H-NMR (400 MHz, MeOH-d4): 1.52-1.70 (2H, m, broad); 1.72-1.84
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(4H, m, broad); 1.98-2.06 (2H, m); 2.45 (3H, s); 2.48-2.54 (2H, m); 3.04-3.10
(4H, m); 3.20-3.24 (2H, m, broad); 6.74 (1H, s, broad); 7.88 (1H, s); 7.28
(1 H, s); 8. 3 7(1 H, s, formate); 8.67 (1 H, s).
Example 22
4-(4-Acetyl-[1, 4]diazepan-1-yl)-N-[5-(6-methoxy-naphthalen-2 yl)-]H-
pyrazol-3 ylJ-butyramide
a) 6-Methoxy-naphthalene-2-carboxylic acid methyl ester
To a solution of 6-methoxy-naphthalene-2-carboxylic acid (1.01 g,
5 mmol) in methanol (10 mL), a catalytic amount of sulphuric acid was added.
The mixture was then heated at 80 C for 8 hours. Upon reaction completion
(as monitored by LcMS), the solution was slowly cooled and the precipitation
of the product was observed. Filtration of the white solid afforded 1.01 g
(94% yield) of title compound.
C13H1203
Mass (calculated) [216]; (found) [M+H+] =217
LC Rt = 2.43, 100% (5 min method)
b) 3-(6-Methoxy-naphthalen-2-yl)-3-oxo propionitrile
To a solution of 6-methoxy-naphthalene-2-carboxylic acid methyl ester
(1.0 g, 4.7 mmol) in dry toluene (8 mL), NaH (0.55 mg, 9.4 mmol) were added
and the mixture was heated at 90 C. To the hot solution, acetonitrile (1.2 mL)
was added dropwise. The reaction was then heated for 18 hours and the
product precipitated from the reaction mixture as its sodium salt.
The reaction was allowed to cool down to room temperature and the
solid formed was first filtered and washed with ether, then it was dissolved
in
water and the solution was acidified with HC1 2N to pH 3, upon which
precipitation of the title compound was observed. Filtration of the solid from
the aqueous solution afforded 1.1 g of title compound (100% of yield).
C13H1203
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Mass (calculated) [225]; (found) [M+H+] =226
LC Rt = 2.13, 90% (5 min method)
c) 5-(6-Methoxy-naphthalen-2 yl)-IH-pyrazol-3-ylamine
To a solution of 3-(6-methoxy-naphthalen-2-yl)-3-oxo-propionitrile
(1.1 g, 4.8 mmoL) in absolute EtOH (10 mL) hydrazine monohydrate
(0.96 mL, 19.2 mmol) was added and the reaction was heated at reflux for
18 hrs. The reaction mixture was allowed to cool to room temperature and the
solvent was evaporated under reduced pressure. The crude was treated with
ether and filtered to afford 0.95 g of title compound (83% of yield).
C1aH13N30
Mass (calculated) [239]; (found) [M+H+] =240
LC Rt = 1.49, 90% (5 min method)
d) 4-(4-Acetyl-[1,4]diazepan-1 yl)-N-[S-(6-methoxy-naphthalen-2 yl)-
1H pyrazol-3 ylJ-butyramide
Following the general method for the synthesis of w-bromo-alkanoic acid
(1H-pyrazol-3-yl-5-aryl)-amides and the general method for the synthesis of w-
amino-alkanoic acid (1H-pyrazol-3-yl-5-aryl)-amides, purification by
preparative
HPLC afforded 15 mg (3% yield) of title compound as its formate salt.
C25H31N503
Mass (calculated) [449]; (found) [M+H+] =450
LC Rt = 1.91, 100% (10 min method)
1H-NMR (400 MHz, MeOH-d4): 1.88-2.0 (4H, m); 2.06 (3H, s);
2.48-2.52 (2H, m); 2.94-3.02 (2H, m); 3.08-3.18 (4H, m); 3.52-3.58 (2H, m);
3.64-3.72 (2H, m); 3.82 (3H, s); 6.78-6.82 (1 H, 'm); 7.04-7.10 (1 H, m);
7.16-7.18 (1H, m); 7.62-7.78 (3H, m); 7.98-8.02 (1H, m); 8.28 (1H, s,
formate).
Example 23
6- (4-Acetyl-[1, 4]diazepan-1 yl)-hexanoic acid [5-(4-methoxy phenyl)-
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IH pyrazol-3 ylJ-amide
The product was prepared according to the general synthetic method for
the one-pot synthesis of co-amino-alkanoic acid (IH-pyrazol-3-yl-5-aryl)-
amides. A solution of 5-bromohexanoyl chloride (0.144 mL, 0.94 mmol) in
dry DMA (1 mL) was cooled to -10 C (ice/water bath) under N2; 5-(4-
methoxy-phenyl)-IH-pyrazol-3-ylamine (178 mg, 0.94 mmol) and
diisopropylethylamine (0.324 mL, 1.88 mmol) were added in dry DMA (1 ml).
The reaction was left stirring for lh at 0 C and then 1-[1,4]diazepan-l-
yl-ethanone (0.310 mL, 2.35 mmol,) and NaI (0.94mmol, 1 eq) were added.
The reaction mixture was heated at 60 C until LC-MS analysis showed
complete conversion of the bromo-intermediate, at which point the reaction
was cooled down and the solvent was removed under reduced pressure. The
residue was dissolved in DCM (2 mL) and washed with saturated Na2CO3
solution.
The organic phase was concentrated under reduced pressure and half of
the crude was purified by Si02 column (gradient from 100% DCM to
DCM-NH3MeOH 2N solution 8:2). The fractions containing the title
compound were collected (35 mg).
C23H33N503
Mass (calculated) [427]; (found) [M+H+] =428
LC Rt = 1.61, 96% (10 min method)
'H-NMR (400 MHz, dmso-d6): 1.24-1.29 (2H, m); 1.36-1.44 (2H, m);
1.54-1.58 (2H, m); 1.62-1.76 (2H, m); 1. 94-1.96 (3H, m); 2.25-2.28 (2H, m);
2.35-2.41 (2H, m); 2.51-2.54 (2H, m); 2.60-2.62 (IH, m); 3.38-3.44 (5H, m);
3.77 (3H, s); 6.73 (1H, s); 6.98 (2H, d, J=8.8 Hz); 7.61 (2H, d, J=8.8); 10.32
(1H, s).
Example 24
N-[S-(4-Methoxy phenyl)-2H pyrazol-3-yl]-2-methyl-4 piperidin-1 yl-
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butyramide
a) Methyl-4-bromo-2-methyl-butyric acid
4-Bromo-2-methyl-butyric acid (2.16 g, 1 eq, prepared according to
the procedure described in J.Am.Chem.Soc. 1990, 112, 2755) was dissolved
in MeOH (10 mL) and a few drops of conc. H2SO4 were added. The reaction
was stirred at reflux for 16 hours. After reaction completion, as monitored by
LC-MS, MeOH was removed under reduced pressure, the oily residue was
diluted with water, the pH adjusted to 9 with 10% NaOH, and the product
was extracted with Et20 (2 X 20 mL) and dried over Na2SO4. The title
compound was obtained as a colourless oil (1.29 g, 55% yield) after solvent
removal.
NMR (400 MHz, CDC13); 1.19 (3H, d); 1.94-1.89 (2H, m); 2.29-2.23
(2H, m); 3.43-3.40 (1H, m); 3.69 (3H, s).
b) 2-Methyl-4 piperidin-1 yl-butyric acid. HCl
15' Methyl-4-bromo-2-methyl-butyric acid (1.29 g, 1 eq) was dissolved in
toluene (15 mL) and piperidine (1.07 mL, 3 eq) was added; the reaction was
stirred for 3 hours. After reaction completion, as monitored by LC-MS,
toluene was removed under reduced pressure and the crude ester was
dissolved in 1 M NaOH (14 mL, 1.1 eq) and MeOH (2 mL). The reaction was
stirred at reflux for 16 hours; after hydrolysis was complete, the reaction
was
concentrated under reduced pressure and the pH adjusted to 4 with 6N HC1.
EtOH was added to help precipitation of NaCI. The organic phase was filtered
and EtOH removed under reduced pressure. The resulting oil was treated with
2M HC1 in Et20 to obtain 2-methyl-4-piperidin-1-yl-butyric acid. HC1 (0.96 g,
66% yield).
CioHi9N02
Mass (calculated) [185.27]; (found) [M+H+]=186.27
LC Rt=0.23, 95% (5 min method)
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c) N-[5-(4-Methoxy phenyl)-2H pyrazol-3 ylJ-2-methyl-4 piperidin-l-
yl-butyramide
2-Methyl-4-piperidin-l-yl-butyric acid. HC1 (0.45 g, 1.2 eq) was
suspended in 1,2-DCE (15 mL) and triethylamine (0.29 mL, 1.2 eq) was
5 added: 1,1'-carbonyldiimidazole (0.303 g, 1.1 eq) was added in one portion
and the reaction was stirred at room temperature for 2 hours. 5-(4-Methoxy-
phenyl)-2H-pyrazol-3-ylamine (0.325 g, 1 eq) was then added and the reaction
stirred at room temperature for further 16 hours. After reaction completion,
as
monitored by LC-MS, the solvent was removed under reduced pressure and
10 the crude amide was purified by column chromatography (Flash-SI 10 g;
CH3CN:MeOH 9:1, CH3CN:2N NH3 MeOH 9:1) to give the title compound as
thick colourless oil (0.120 g, 0.33 mmol).
C2oH28Na02
Mass (calculated) [356.48]; (found) [M+H+]=357.25
15 LC Rt=1.67, 97% (10 min method)
NMR (400 MHz, dmso-d6); 1.18 (3H, d); 1.35-1.31 (2H, m); 1.46-
1.41 (4H, m); 1.77-1.72 (1 H, m); 2.19-2.16 (2H, m); 2.27-2.23 (4H, m);
2.61-2.58 (2H, m); 3.76 (3H, s); 6.76 (1H, s); 6.92 (2H, d); 7.61 (2H, d);
10.33 (1H, s).
20 Example 25
N-[4-(4-Methoxy phenyl)-]H-imidazol-2 ylJ-4 piperidin-1 yl-
butyramide
To a suspension of 4-piperidin-1-yl-butyric acid (200 mg, 1.17 mmol,
1.0 eq) in 1,2-dichloroethane (2 mL), N,N'-carbonyldiimidazole (179.9 mg,
25 1.11 mmol, 0.95 eq) was added and the mixture was stirred at room
temperature for 1 hour until complete activation of the aminoacid and
dissolution of the suspension. 4-(4-Methoxy-phenyl)-1 H-imidazol-2-ylamine
(prepared according to the procedure reported in JOC 1994, 59, 24, 7299;
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110.5 g, 0.58 mmol, 0.50 eq) was added and the reaction stirred for 1 day at
50 C. The slow conversion was monitored by LC-MS. Another aliquote of
activated acid (4-piperidin-1-yl-butyric acid, 200 mg and carbonyldiimidazole,
179.9 mg in 2 mL of 1,2-dichloroethane) were added and the reaction stirred
for further two days at 50 C.
The solvent was evaporated under reduced pressure and the crude
mixture purified by preparative HPLC to obtain a 9:1 mixture of the product
and unreacted 4-(4-methoxy-phenyl)-1 H-imidazol-2-ylamine. The crude was
purified by treatment with isocyanate resin and SCX column to give 78.0 mg
(Yield: 39%) of the title compound as a white solid.
C19H26N402 Mass (calculated) [342]; (found) [M+H+] =343
LC Rt = 1.00 (and solvent front), 99% (10 min method)
'H-NMR (400 MHz, DMSO): 1.30-1.36 (2H, m); 1.43-1.49 (4H, m);
1.67-1.75 (2H, m); 2.22-2.34 (8H, m); 3.73 (3H, s, -OCH3); 6.87 (2H, d, J=8.8
Hz); 7.10 (1 H, s); 7.60 (2H, d, J= 8.8 Hz); 11.26 (1 H, s, NHCO), 11.52 (1H,
s,
NH).
13C-NMR (400 MHz, DMSO): 21.54 (1 C); 23.63 (1 C); 24.92
(2C); 33.24 (1C); 53.6 (1C, -OCH3); 55.02 (2C); 57.46 (1C); 113.88 (2C);
125.18 (2C), 141.13 (1 C); 157.67 (1 C); 162.33 (2C); 163.66 (1 C); 171.15
(1 C, CO).
Table 1- Examples 26-171
Table 1 shows a selection of the compounds synthesised, which were
prepared according to the method indicated in the last column of the table and
discussed in detail in the Experimental Procedures with the synthesis of
Examples 1-25.
When the compound is indicated as the HC1 salt, the salt was formed
by dissolution of the free base in methanol and addition of 1 eq 1M HC1 in
ether followed by evaporation of the solvents. When the compound is
CA 02637530 2008-07-17
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indicated as HCOOH (formic acid) salt, the compound was purified by
preparative HPLC.
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Biological activity
Cloning of alpha7 nicotinic acetylcholine receptor and generation of
stable recombinant alpha7 nAChR expressing cell lines
Full length cDNAs encoding the alpha7 nicotinic acetylcholine receptor
were cloned from a rat brain cDNA library using standard molecular biology
techniques. Rat GH4C 1 cells were then transfected with the rat receptor,
cloned and analyzed for functional alpha7 nicotinic receptor expression
employing a FLIPR assay to measure changes in intracellular calcium
concentrations. Cell clones showing the highest calcium-mediated
fluorescence signals upon agonist (nicotine) application were further
subcloned and subsequently stained with Texas red-labelled a-bungarotoxin
(BgTX) to analyse the level and homogeneity of alpha7 nicotinic
acetylcholine receptor expression using confocal microscopy. Three cell lines
were then expanded and one characterised pharmacologically (see Table 2
below) prior to its subsequent use for compound screening.
Table 2 - Pharmacological characterisation of alpha7 nAChR stably
expressed in GH4C1 cells using the functional FLIPR assay
Compound EC50 [microM]
Acetylcholine 3.05 0.08 (n=4)
Choline 24.22 8.30 (n=2)
Cytisine 1.21 0.13 (n=5)
DMPP 0.98 0.47 (n=6)
Epibatidine 0.0 12 0.002 (n=7)
Nicotine 1.03 0.26 (n=22)
Development of a functional FLIPR assay for primary screening and
concentration-response analysis
A robust functional FLIPR assay (Z' = 0.68) employing the stable
recombinant GH4C 1 cell line was developed to screen the alpha7 nicotinic
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134
acetylcholine receptor. The FLIPR system allows the measurements of real
time Ca2+-concentration changes in living cells using a Ca2+ sensitive
fluorescence dye (such as Fluo4). This instrument enables the screening for
agonists and antagonists for alpha 7 nAChR channels stably expressed in
GH4C 1 cells.
Cell culture
GH4C 1 cells stably transfected with rat- alpha7-nAChR (see above)
were used. These cells are poorly adherent and therefore pretreatment of
flasks and plates with poly-D-lysine was carried out. Cells are grown in
150 cm2 T-flasks, filled with 30m1 of medium at 37 C and 5% CO2.
Data analysis
EC50 and IC50 values were calculated using the IDBS XLfit4.1 software
package employing a sigmoidal concentration-response (variable slope)
equation:
Y= Bottom + ((Top-Bottom)/(1+((EC50/X) ^HillSlope))
Assay validation
The functional FLIPR assay was validated with the alpha7 nAChR
agonists nicotine, cytisine, DMPP, epibatidine, choline and acetylcholine.
Concentration-response curves were obtained in the concentration range from
0.001 to 30 microM. The resulting EC50 values are listed in Table 2 and the
obtained rank order of agonists is in agreement with published data (Quik et
al., 1997)(22).
The assay was further validated with the specific alpha7 nAChR
antagonist MLA (methyllycaconitine), which was used in the concentration
range between 1 microM to 0.01 nM, together with a competing nicotine
concentration of 10 microM. The IC50 value was calculated as 1.31+0.43 nM
in nine independent experiments.
Development offunctional FLIPR assays for selectivity testing
Functional FLIPR assays were developed in order to test the selectivity
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135
of compounds against the alphal (muscular) and alpha3 (ganglionic) nACh
receptors and the structurally related 5-HT3 receptor. For determination of
activity at alphal receptors natively expressed in the rhabdomyosarcoma
derived TE 671 cell line an assay employing membrane potential sensitive
dyes was used, whereas alpha3 selectivity was determined by a calcium-
monitoring assays using the native SH-SY5Y cell line. In order to test
selectivity against the 5-HT3 receptor, a recombinant cell line was
constructed
expressing the human 5-HT3A receptor in HEK 293 cells and a
calcium-monitoring FLIPR assay employed.
Screening of compounds
The compounds from Examples 1-171 described showed agonist
activity in the functional FLIPR primary screening assay employing the stable
recombinant GH4C 1 cell line expressing the alpha7 nAChR. The most potent
hits identified were validated further by generation of concentration-response
curves. The potency of compounds from Examples 1-153 as measured in the
functional FLIPR screening assay was found to range between 10 nM and
30 microM, with the majority showing a potency ranging between 10 nM and
10 microM.
The best exemplified compounds were also demonstrated to be selective
against the alphal nACh, alpha3 nACh and 5HT3 receptors.
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136
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