U.S. patent application number 11/195747 was filed with the patent office on 2005-12-01 for dipeptide nitrile cathepsin k inhibitors.
This patent application is currently assigned to NORVARTIS AG. Invention is credited to Missbach, Martin.
Application Number | 20050267129 11/195747 |
Document ID | / |
Family ID | 9885359 |
Filed Date | 2005-12-01 |
United States Patent
Application |
20050267129 |
Kind Code |
A1 |
Missbach, Martin |
December 1, 2005 |
Dipeptide nitrile cathepsin K inhibitors
Abstract
Dipeptide nitrile Cathepsin K inhibitors of formula I, and
pharmaceutically acceptable salts or esters thereof 1 In which
R.sub.1 and R.sub.2 are independently H or C.sub.1-C.sub.7lower
alkyl, or R.sub.1 and R.sub.2 together with the carbon atom to
which they are attached form a C.sub.3-C.sub.8cycloalkyl ring, and
Het is an optionally substituted nitrogen-containing heterocyclic
substituent, are provided, useful e.g. for therapeutic or
prophylactic treatment of a disease or medical condition in which
cathepsin K is implicated.
Inventors: |
Missbach, Martin;
(Gipf-Oberfrick, CH) |
Correspondence
Address: |
OBLON, SPIVAK, MCCLELLAND, MAIER & NEUSTADT, P.C.
1940 DUKE STREET
ALEXANDRIA
VA
22314
US
|
Assignee: |
NORVARTIS AG
Basel
CH
|
Family ID: |
9885359 |
Appl. No.: |
11/195747 |
Filed: |
August 3, 2005 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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11195747 |
Aug 3, 2005 |
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10410377 |
Apr 9, 2003 |
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10410377 |
Apr 9, 2003 |
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09778302 |
Feb 7, 2001 |
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6642239 |
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Current U.S.
Class: |
514/255.03 ;
514/317; 544/396; 546/229 |
Current CPC
Class: |
A61P 19/00 20180101;
A61P 21/04 20180101; A61P 29/00 20180101; C07D 295/155 20130101;
A61P 1/02 20180101; A61P 9/10 20180101; A61P 3/14 20180101; C07D
211/34 20130101; A61P 19/10 20180101; A61P 43/00 20180101; A61P
33/00 20180101; A61P 35/00 20180101; A61P 31/00 20180101; Y02A
50/423 20180101; Y02A 50/411 20180101; A61P 9/00 20180101; A61P
11/00 20180101; Y02A 50/414 20180101; A61P 33/06 20180101; A61P
19/02 20180101; A61P 37/06 20180101; C07C 2601/14 20170501 |
Class at
Publication: |
514/255.03 ;
514/317; 546/229; 544/396 |
International
Class: |
A61K 031/495; A61K
031/445; C07D 211/26 |
Foreign Application Data
Date |
Code |
Application Number |
Feb 10, 2000 |
GB |
0003111.2 |
Claims
1-9. (canceled)
10. A compound of formula I, or a pharmaceutically acceptable salt
or ester thereof 6wherein R.sub.1 and R.sub.2 are independently H
or C.sub.1-C.sub.7lower alkyl; or R.sub.1 and R.sub.2 together with
the carbon atom to which they are attached form a
C.sub.3-C.sub.8cycloalkyl ring; and Het is a saturated
nitrogen-containing heterocyclic ring containing at least one
nitrogen atom and from 2 to 10 carbon atoms, optionally substituted
at nitrogen by C.sub.1-C.sub.7lower alkyl, C.sub.1-C.sub.7lower
alkoxy-C.sub.1-C.sub.7lower alkyl,
C.sub.5-C.sub.10aryl-C.sub.1-C.sub.7lower alkyl or
C.sub.3-C.sub.8cycloalkyl.
11. A compound according to claim 10 of formula II, or a
pharmaceutically acceptable salt or ester thereof 7wherein X is CH
or N, and R is C.sub.1-C.sub.7lower alkyl, C.sub.1-C.sub.7lower
alkoxy-C.sub.1-C.sub.7lo- wer alkyl,
C.sub.5-C.sub.10aryl-C.sub.1-C.sub.7lower alkyl, or
C.sub.3-C.sub.8cycloalkyl.
12. A pharmaceutical composition comprising a compound according to
claim 10 as an active ingredient.
13. A compound according to claim 10, wherein the saturated
nitrogen-containing heterocyclic ring system contains from 3 to 7
carbon atoms.
14. A compound according to claim 10, wherein the saturated
nitrogen-containing heterocyclic ring system contains 4 or 5 carbon
atoms.
15. A compound according to claim 10, wherein R.sub.1 and R.sub.2
are H.
16. A compound according to claim 10, in which the saturated
nitrogen-containing heterocyclic ring is optionally substituted at
nitrogen by C.sub.1-C.sub.7lower alkyl.
17. A compound according to claim 10, in which the saturated
nitrogen-containing heterocyclic ring is substituted at nitrogen by
C.sub.1-C.sub.7lower alkyl.
18. A compound according to claim 10, in which the saturated
nitrogen-containing heterocyclic ring is substituted at nitrogen by
C.sub.1-C.sub.7lower alkoxy-C.sub.1-C.sub.7lower alkyl.
19. A compound according to claim 10, in which the saturated
nitrogen-containing heterocyclic ring is substituted at nitrogen by
C.sub.5-C.sub.10aryl-C.sub.1-C.sub.7lower alkyl.
20. A compound according to claim 10, in which the saturated
nitrogen-containing heterocyclic ring is substituted at nitrogen by
C.sub.3-C.sub.8cycloalkyl.
21. A compound according to claim 10, wherein Het is piperazinyl or
piperidinyl each of which is optionally substituted at nitrogen by
C.sub.1-C.sub.7lower alkyl, C.sub.1-C.sub.7lower
alkoxy-C.sub.1-C.sub.7lo- wer alkyl,
C.sub.5-C.sub.10aryl-C.sub.1-C.sub.7lower alkyl or
C.sub.3-C.sub.8cycloalkyl.
22. A compound according to claim 21, in which Het is
piperazin-1-yl or piperidin-4-yl optionally substituted at nitrogen
by C.sub.1-C.sub.7lower alkyl.
23. A compound according to claim 21, which is
N-[1-cyanomethyl-carbamoyl)-
-cyclohexyl]-4-(4-methyl-piperazin-1-yl)-benzamide.
24. A method of treating a mammal suffering from a disease or
medical condition which involves elevated levels of cathepsin K,
such disease or medical condition being selected from hypercalcemia
of malignancy, excessive bone loss, excessive cartilage degradation
and excessive matrix degradation, comprising administering to said
mammal an effective cathepsin K inhibiting amount of a compound
according to claim 10.
25. A method of treating osteoporosis in a mammal comprising
administering to said mammal an effective amount of a compound
according to claim 10.
26. A method of treating rheumatoid arthritis in a mammal
comprising administering to said mammal an effective amount of a
compound according to claim 10.
27. A method of treating osteoarthritis in a mammal comprising
administering to said mammal an effective amount of a compound
according to claim 10.
28. A method of treating gingival diseases in a mammal comprising
administering to said mammal an effective amount of a compound
according to claim 10.
29. A method of treating Paget's disease in a mammal comprising
administering to said mammal an effective amount of a compound
according to claim 10.
30. A method of treating hypercalcemia of malignancy in a mammal
comprising administering to said mammal an effective amount of a
compound according to claim 10.
31. A method of selectively inhibiting cathepsin K activity in a
mammal with elevated levels of cathepsin K which comprises
administering to a mammal in need thereof an effective cathepsin K
inhibiting amount of a compound according to claim 10.
Description
[0001] This invention relates to inhibitors of cysteine proteases,
in particular to dipeptide nitrile cathepsin K inhibitors and to
their pharmaceutical use for the treatment or prophylaxis of
diseases or medical conditions in which cathepsin K is
implicated.
[0002] Cathepsin K is a member of the family of lysosomal cysteine
cathepsin enzymes, e.g. cathepsins B, K, L and S, which are
implicated in various disorders including inflammation, rheumatoid
arthritis, osteoarthritis, osteoporosis, tumors (especially tumor
invasion and tumor metastasis), coronary disease, atherosclerosis
(including atherosclerotic plaque rupture and destabilization),
autoimmune diseases, respiratory diseases, infectious diseases and
immunologically mediated diseases (including transplant
rejection).
[0003] Our copending International patent application WO 99/24460
describes dipeptide nitrites which are inhibitors of cysteine
cathepsins and their use for treatment of cysteine cathepsin
dependent diseases or medical conditions. New dipeptide nitrile
compounds have now been made which are inhibitors of cathepsin K,
and which have desirable properties for pharmaceutical
applications.
[0004] Accordingly the present invention provides a compound of
formula I, or a pharmaceutically acceptable salt or ester thereof
2
[0005] In which R.sub.1 and R.sub.2 are independently H or
C.sub.1-C.sub.7lower alkyl, or R.sub.1 and R.sub.2 together with
the carbon atom to which they are attached form a
C.sub.3-C.sub.8cycloalkyl ring, and Het is an optionally
substituted nitrogen-containing heterocyclic substituent, provided
that Het is not 4-pyrrol-1-yl.
[0006] The Het substituent may be at the 2- or 3-position of the
phenyl ring, though is preferably at the 4-position.
[0007] In the present description "nitrogen-containing heterocycle"
signifies a heterocyclic ring system containing at least one
nitrogen atom, from 2 to 10, preferably from 3 to 7, most
preferably 4 or 5, carbon atoms and optionally one or more
additional heteroatoms selected from O, S or preferably N.
[0008] Het may comprise an unsaturated, e.g. an aromatic,
nitrogen-containing heterocycle; though preferably comprises a
saturated nitrogen-containing heterocycle. Particularly preferred
saturated nitrogen-containing heterocycles are piperazinyl,
preferably piperazin-1-yl or piperidinyl, preferably
piperidin-4-yl.
[0009] Het may be substituted by one or more substituents, e.g. by
up to 5 substituents independently selected from halogen, hydroxy,
amino, nitro, optionally substituted C.sub.1-4alkyl (e.g. alkyl
substituted by hydroxy, alkyloxy, amino, optionally substituted
alkylamnino, optionally substituted dialkylarnino, aryl or
heterocyclyl), C.sub.1-4alkoxy.
[0010] Preferably Het is substituted at a nitrogen atom, most
preferably mono-substituted at a nitrogen atom.
[0011] Preferred substituents for Het are C.sub.1-C.sub.7lower
alkyl, C.sub.1-C.sub.7lower alkoxy-C.sub.1-C.sub.7lower alkyl,
C.sub.5-C.sub.10aryl-C.sub.1-C.sub.7lower alkyl, or
C.sub.3-C.sub.8cycloalkyl.
[0012] R.sub.1 and R.sub.2 as C.sub.1-C.sub.7lower alkyl are
preferably the same, e.g. methyl, or R.sub.1 and R.sub.2 together
with the carbon atom to which they are attached preferably form a
C.sub.3-C.sub.8cycloalk- yl ring, e.g. a cyclopropyl ring. Most
preferably both R.sub.1 and R.sub.2 are H.
[0013] Thus in particularly preferred embodiments the invention
provides a compound of formula II, or a pharmaceutically acceptable
salt or ester thereof 3
[0014] wherein X is CH or N, and R is H, C.sub.1-C.sub.7lower
alkyl, C.sub.1-C.sub.7lower alkoxy-C.sub.1-C.sub.7lower alkyl,
C.sub.5-C.sub.10aryl-C.sub.1-C.sub.7lower alkyl, or
C.sub.3-C.sub.8cycloalkyl.
[0015] Thus particular examples of R as C.sub.1-C.sub.7lower alkyl
are methyl, ethyl, n-propyl, or i-propyl.
[0016] A particular example of R as C.sub.1-C.sub.7lower
alkoxy-C.sub.1-C.sub.7lower alkyl is methoxyethyl.
[0017] A particular example of R as
C.sub.5-C.sub.10aryl-C.sub.1-C.sub.7lo- wer alkyl is benzyl.
[0018] A particular example of R as C.sub.3-C.sub.8cycloalkyl is
cyclopentyl.
[0019] Examples of particular compounds of formula II are:
[0020]
N-[1-(Cyanomethyl-carbamoyl)-cyclohexyl]-4-(piperazin-1-yl)-benzami-
de;
[0021]
N-[1-(Cyanomethyl-carbamoyl)-cyclohexyl]-4-(4-methyl-piperazin-1-yl-
)-benzamide;
[0022]
N-[1-(Cyanomethyl-carbamoyl)-cyclohexyl]-4-(4-ethyl-piperazin-1-yl)-
-benzamide;
[0023]
N-[1-(Cyanomethyl-carbamoyl)-cyclohexyl]-4-[4-(1-propyl)-piperazin--
1-yl]-benzamide;
[0024]
N-[1-(Cyanomethyl-carbamoyl)-cyclohexyl]-4-(4-isopropyl-piperazin-1-
-yl)-benzamide;
[0025]
N-[1-(Cyanomethyl-carbamoyl)-cyclohexyl]-4-(4-benzyl-piperazin-1-yl-
)-benzamide;
[0026]
N-[1-(Cyanomethyl-carbamoyl)-cyclohexyl]-4-[4-(2-methoxy-ethyl)-pip-
erazin-1-yl]-benzamide;
[0027]
N-[1-(Cyanomethyl-carbamoyl)-cyclohexyl]-4-(1-propyl-piperidin-4-yl-
)-benzamide;
[0028]
N-[1-(Cyanomethyl-carbamoyl)-cyclohexyl]-4-[1-(2-methoxy-ethyl)-pip-
eridin-4-yl]-benzamide;
[0029]
N-[1-(Cyanomethyl-carbamoyl)-cyclohexyl]-4-(1-isopropyl-piperidin-4-
-yl)-benzamide;
[0030]
N-[1-(Cyanomethyl-carbamoyl)-cyclohexyl]-4-(1-cyclopentyl-piperidin-
4-yl)-benzamide;
[0031]
N-[1-(Cyanomethyl-carbamoyl)-cyclohexyl]4-(1-methyl-piperidin-4-yl)-
-benzamide, and
[0032]
N-[1-(Cyanomethyl-carbamoyl)-cyclohexyl]4-(piperidin-4-yl)-benzamid-
e.
[0033] Compounds of formula I and II and the specific compounds
above are hereinafter referred to as Compounds of the
Invention.
[0034] Compounds of the Invention may be prepared by coupling the
corresponding Het substituted benzoic acid derivative with
1-amino-cyclohexanecarboxylic acid cyanomethyl amide. For example,
the benzoic acid derivative, preferably in the form of its
hydrochloride, is mixed with 1-amino-cyclohexanecarboxylic acid
cyanomethyl amide, e.g. in the presence of HOBT
(1-hydroxybenzotriazole), WSCD and triethylamine, in solution, e.g.
in DMF, and stirred, e.g. overnight at room temperature. The
product may be recovered, for instance, by evaporation of the
solvent, followed by washing with aqueous sodium carbonate
solution, preferably under mildly basic conditions, followed by
solvent extraction, e.g. with ethyl acetate, drying of the extract,
e.g. over sodium sulfate, evaporation of the solvent and
filtration. Alternative procedures and reagents may be used; for
instance, as hereinafter described in the Examples.
[0035] Thus in a further aspect the invention provides a process
for the preparation of a compound of formula I which comprises
coupling the corresponding Het substituted benzoic acid derivative
of formula III 4
[0036] With 1-amino-cyclohexanecarboxylic acid cyanomethyl-amide.
5
[0037] 1-Amino-cyclohexanecarboxylic acid cyanomethyl-amide may be
prepared by coupling 1-amino-cyclohexane carboxylic acid, typically
in appropriate amino protected form, e.g. FMOC-1-amino-cyclohexane
carboxylic acid, with 2-aminoacetonitrile. For example,
FMOC-1-amino-cyclohexane carboxylic acid, e.g. with HOBT and WSCD,
is added to a solution of 2-aminoacetonitrile and triethylamine in
DMF and the mixture stirred at 25.degree. C. overnight.
1-Amino-cyclohexanecarbox- ylic acid cyanomethyl-amide may be
recovered as described in the Examples. FMOC-1-amino-cyclohexane
carboxylic acid may be prepared as described in the Examples.
[0038] Compounds of the invention are either obtained in the free
form, or as a salt thereof if salt forming groups are present.
[0039] Compounds of the Invention having basic groups can be
converted into acid addition salts, especially pharmaceutically
acceptable salts. These are formed, for example, with inorganic
acids, such as mineral acids, for example sulfuric acid, a
phosphoric or hydrohalic acid, or with organic carboxylic acids,
such as (C.sub.1-C.sub.4)alkanecarboxylic acids which, for example,
are unsubstituted or substituted by halogen, for example acetic
acid, such as saturated or unsaturated dicarboxylic acids, for
example oxalic, succinic, maleic or fumaric acid, such as
hydroxycarboxylic acids, for example glycolic, lactic, malic,
tartaric or citric acid, such as amino acids, for example aspartic
or glutamic acid, or with organic sulfonic acids, such as
(C.sub.1-C.sub.4)-alkylsulfonic acids (for example methanesulfonic
acid) or arylsulfonic acids which are unsubstituted or substituted
(for example by halogen). Preferred are salts formed with
hydrochloric acid, methanesulfonic acid and maleic acid.
[0040] In view of the close relationship between the free compounds
and the compounds in the form of their salts, whenever a compound
is referred to in this context, a corresponding salt is also
intended, provided such is possible or appropriate under the
circumstances.
[0041] The compounds, including their salts, can also be obtained
in the form of their hydrates, or include other solvents used for
their crystallization.
[0042] The compounds of the invention exhibit valuable
pharmacological properties in mammals and are particularly useful
as inhibitors of cathepsin K.
[0043] The cathepsin K inhibitory effects of the compound of the
invention can be demonstrated in vitro by measuring the inhibition
of e.g. recombinant human cathepsin K.
[0044] The in vitro Assay is Carried Out as Follows:
[0045] For cathepsin K:
[0046] The assay is performed in 96 well microtiter plates at
ambient temperature using recombinant human cathepsin K. Inhibition
of cathepsin K is assayed at a constant enzyme (0.16 nM) and
substrate concentration (54 mM Z-Phe-Arg-AMCA-Peptide Institute
Inc. Osaka, Japan) in 100 mM sodium phosphate buffer, pH 7.0,
containing 2 nM dithiothreitol, 20 mM Tween 80 and 1 mM EDTA.
Cathepsin K is preincubated with the inhibitors for 30 min, and the
reaction is initiated by the addition of substrate. After 30 min
incubation the reaction is stopped by the addition of E-64 (2 mM),
and fluorescence intensity is read on a multi-well plate reader at
excitation and emission wavelengths of 360 and 460 nm,
respectively. Compounds of the Invention typically have Kis for
human cathepsin K of less than about 50 nM, preferably of about 5
nM or less, e.g. about 1 nM.
[0047] In view of their activity as inhibitors of cathepsin K,
Compounds of the Invention are particularly useful in mammals as
agents for treatment and prophylaxis of diseases and medical
conditions involving elevated levels of cathepsin K. Such diseases
include diseases involving infection by organisms such as
pneumocystis carinii, trypsanoma cruzi, trypsanoma brucei,
crithidia fusiculata, as well as parasitic diseases such as
schistosomiasis and malaria, tumours (tumour invasion and tumour
metastasis), and other diseases such as metachromatic
leukodystrophy, muscular dystrophy, amytrophy and similar
diseases.
[0048] Cathepsin K, has been implicated in diseases of excessive
bone loss, and thus the Compounds of the Invention may be used for
treatment and prophylaxis of such diseases, including osteoporosis,
gingival diseases such as gingivitis and periodontitis, Paget's
disease, hypercalcemia of malignancy, e.g. tumour-induced
hypercalcemia and metabolic bone disease. Also the Compounds of the
Invention may be use for treatment or prophylaxis of diseases of
excessive cartilage or matrix degradation, including osteoarthritis
and rheumatoid arthritis as well as certain neoplastic diseases
involving expression of high levels of proteolytic enzymes and
matrix degradation.
[0049] Compounds of the Invention, are also indicated for
preventing or treating coronary disease, atherosclerosis (including
atherosclerotic plaque rupture and destabilization), autoimmune
diseases, respiratory diseases and immunologically mediated
diseases (including transplant rejection).
[0050] Compounds of the Invention are particularly indicated for
preventing or treating osteoporosis of various genesis (e.g.
juvenile, menopausal, post-menopausal, post-traumatic, caused by
old age or by cortico-steroid therapy or inactivity).
[0051] Beneficial effects are evaluated in in vitro and in vivo
pharmacological tests generally known in the art, and as
illustrated herein.
[0052] The above cited properties are demonstrable in in vitro and
in vivo tests, using advantageously mammals, e.g. rats, mice, dogs,
rabbits, monkeys or isolated organs and tissues, as well as
mammalian enzyme preparations, either natural or prepared by e.g.
recombinant technology. Compounds of the Invention can be applied
in vitro in the form of solutions, e.g. preferably aqueous
solutions or suspensions, and in vivo either enterally or
parenterally, advantageously orally, e.g. as a suspension or in
aqueous solution, or as a solid capsule or tablet formulation. The
dosage in vitro may range between about 10.sup.-5 molar and
10.sup.-9 molar concentrations. The dosage in vivo may range,
depending on the route of administration, between about 0.1 and 100
mg/kg.
[0053] In accordance with the present invention it has been found
that Compounds of the Invention, have good bioavailability, in
particular good oral bioavailability. Thus, for example selected
compounds of the Invention have absolute oral bioavailabilities of
50% or greater e.g. about 80% or more.
[0054] The antiarthritic efficacy of the Compounds of the Invention
for the treatment of rheumatoid arthritis can be determined using
models such as or similar to the rat model of adjuvant arthritis,
as described previously (R. E. Esser, et. al. J. Rheumatology,
1993, 20, 1176.)
[0055] The efficacy of the compounds of the invention for the
treatment of osteoarthritis can be determined using models such as
or similar to the rabbit partial lateral meniscectomy model, as
described previously (Colombo et al. Arth. Rheum. 1993 26,
875-886). The efficacy of the compounds in the model can be
quantified using histological scoring methods, as described
previously (O'Byrne et al. Inflamm Res 1995, 44, S117-S118).
[0056] The efficacy of the compounds of the invention for the
treament of osteoporosis can be determined using an animal model
such as the ovariectomised rat or other similar species, e.g.
rabbit or monkey, in which test compounds are administered to the
animal and the presence of markers of bone resorption are measured
in urine or serum (e.g. as described in Osteoporos Int (1997)
7:539-543).
[0057] Accordingly in further aspects the invention provides: A
Compound of the Invention for use as a pharmaceutical; a
pharmaceutical composition comprising a Compound of the Invention
as an active ingredient; a method of treating a patient suffering
from or susceptible to a disease or medical condition in which
cathepsin K is implicated, comprising administering an effective
amount of a Compound of the Invention to the patient, and the use
of a Compound of the Invention for the preparation of a medicament
for therapeutic or prophylactic treatment of a disease or medical
condition in which cathepsin K is implicated.
[0058] The present invention relates to methods of using Compounds
of the Invention and their pharmaceutically acceptable salts, or
pharmaceutical compositions thereof, in mammals for inhibiting
cathepsin K, and for the treatment of cathepsin K dependent
conditions, such as the cathepsin K dependent conditions, described
herein, e.g. inflammation, osteoporosis, rheumatoid arthritis and
osteoarthritis.
[0059] Particularly the present invention relates to a method of
selectively inhibiting cathepsin K activity in a mammal which
comprises administering to a mammal in need thereof an effective
cathepsin K inhibiting amount of a Compound of the Invention.
[0060] More specifically such relates to a method of treating
osteoporosis, rheumatoid arthritis, osteoarthritis, and
inflammation (and other diseases as identified above) in mammals
comprises administering to a mammal in need thereof a
correspondingly effective amount of a Compound of the
Invention.
[0061] The following examples are intended to illustrate the
invention and are not to be construed as being limitations thereon.
Temperatures are given in degrees Centrigrade. If not mentioned
otherwise, all evaporations are performed under reduced pressure,
preferably between about 15 and 100 mm Hg (=20-133 mbar). The
structure of final products, intermediates and starting materials
is confirmed by standard analytical methods, e.g. microanalysis and
spectroscopic characteristics (e.g. MS, IR, NMR). Abbreviations
used are those conventional in the art.
EXAMPLES
Synthesis of 1-Amino-cyclohexanecarboxylic acid
cyanomethyl-amide
A. FMOC-1-aminocyclohexane carboxylic acid
[0062] The title compound is prepared from 1-aminocyclohexane
carboxylic acid (700 mmol), FMOC-chloride (770 mmol),
Diisopropyl-ethylamine (770 mmol) and 770 ml NaOH 1N in 950 ml
dioxan by conventional methods. Mp. 180-182.degree. C.; Rf=0.21
(CH2Cl2/MeOH=95:5)
[0063] Acetonitrile may be used as solvent in place of dioxan.
B. FMOC-1-amino-cyclohexanecarboxylic acid cyanomethyl-amide
[0064] 2-Aminoacetonitril hydrochloride (564 mmol) and
triethylamine (564 mmol) are dissolved in DMF (1700 ml).
FMOC-1-aminocyclohexane carboxylic acid (564 mmol), HOBt (564 mmol)
and WSCD (564 mmol) are added and the mixture is stirred at
25.degree. C. overnight. After evaporation of the solvent, the
residue is dissolved in a mixture of water and sodium carbonate (to
ensure slightly basic conditions) and extracted three times with
ethyl acetate. The extract is washed with water, 10% citric acid,
brine, sodium bicarbonate, brine and dried over magnesium sulfate
and evaporated. The residue is suspended in diethylether and the
solid filtered of and dried (vacuum). A white powder with mp.
167-169.degree. C., Rf=0.27 (n-hexane:ethyl acetate=1:1) is
obtained.
[0065] Alternatively THF may be used as the solvent and
1-chloro-3,5-dimethoxytriazine (CDMT) as the activator, together
with N-methylmorpholine (NMM) during the coupling reaction; in
which case the product may be recovered by addition of
isopropylacetate and water, separation of the organic phase
followed by washing with brine, partial evaporation of the solvent,
recovery of the crystallised product by filtration and drying.
C. 1-Amino-cyclohexanecarboxylic acid cyanomethyl-amide
[0066] FMOC-1-amino-cyclohexanecarboxylic acid cyanomethyl-amide
(248 mmol) is dissolved in DMF (200 ml), piperidine (248 mmol) is
added and the mixture is stirred at RT for 2 hours. The reaction
mixture is poured into water (3000 ml) and stirred for 30 minutes.
The suspension is filtered and the filtrate is acidified with HCl
4N and than extracted with ethyl acetate. NaOH 1N is added to make
the water phase basic and the mixture is extracted three times with
ethyl acetate. The organic fractions are dried over sodium sulfate
and the solvent is evaporated. The residue is dried (vacuum) to
yield a pale yellow oil. Rf=0.26 (CH2Cl2/MeOH=95:5).
[0067] 1H-NMR (d6-DMSO): 1.05-1.80 (m, 10 H); 4.0 (br. s, 2H); NH
very broad signal.
[0068] Alternatively THF may be used in place of DMF and
diethylamine inplace of piperidine in the the FMOC deprotection
step.
Example 1
Synthesis of
N-[1-(Cyanomethyl-carbamoyl)-cyclohexyl]-4-piperazin-1-yl-ben-
zamide
A. 4-piperazin-1-yl-benzoic acid methyl ester
[0069] 1-(4-Cyanophenyl)-piperazine (11 mmol) is dissolved in 15 ml
of a mixture of concentrated sulfonic acid and methanol (5N) and
stirred in a sealed tube at 110.degree. C. for 3 hours. After
evaporation of the solvent, the residue is dissolved in water and
extracted with ethyl acetate. Addition of sodium carbonate to the
water phase until pH=9 results in the precipitation of a white
solid which is filtered off and dried (vacuum). A white powder with
Rf=0.59 (CH.sub.2Cl.sub.2/MeOH (+NH.sub.3 3N)=9:1) is obtained.
B. 4-piperazin-1-yl-benzoic acid hydrochlorid
[0070] 4-piperazin-1-yl-benzoic acid methyl ester (17 mmol) is
dissolved in 6N HCl (25 ml) and heated under reflux for 3 hours.
The mixture is cooled in an ice bath to 0-4.degree. C. and the
solid material formed is filtered off, washed with acetone and
dried (vacuum). A white powder with mp. >240.degree. C. is
obtained.
C. 4-(4-FMOC-piperazin-1-yl)-benzoic acid
[0071] 4-Piperazin-1-yl-benzoic acid hydrochlorid (10.5 mmol) is
dissolved in 15 ml Dioxan and 11.6 ml NaOH (2N) and cooled to
0.degree. C. Simultaneously, FMOC-chloride (11.6 mmol) in dioxan (5
ml) and diisopropyl-ethylamine (11.6 mmol) in dioxan (5 ml) are
added dropwise over 20 minutes at 0.degree. C. and the mixture is
stirred for 15 minutes and is then allowed to warm up to rt and is
stirred over night. The mixture is diluted with water (50 ml) and
extracted 2 times with diethylether. The water phase is acidified
with aqueous HCl (4N) at 0-4.degree. C. and the solid material
formed is filtered off, washed with water and dried (vacuum). A
white powder with Rf=0.2 (CH.sub.2Cl.sub.2/MeOH=95:5) is
obtained.
D.
N-[1-(Cyanomethyl-carbamoyl)-cyclohexyl]-4-(4-FMOC-piperazin-1-yl)-benz-
amide
[0072] 1-Amino-cyclohexanecarboxylic acid cyanomethyl-amide (8.3
mmol) 4-(4-FMOC-piperazin-1-yl)-benzoic acid (8.3 mmol), HOBT (8.3
mmol) and WSCD (8.3 mmol) are dissolved in DMF (20 ml) and stirred
overnight at rt. After evaporation of the solvent, the residue is
dissolved in a mixture of water and sodium carbonate (to ensure
slightly basic conditions) and extracted three times with ethyl
acetate. The combined extract is dried over sodium sulfate and
evaporated. The residue is purified by flash chromatography on
silica gel with (ethylacetate/hexane=4:1) as mobile phase. The
product containing fractions are combined and evaporated. The
residue is suspended in diethylether and the solid filtered of and
dried (vacuum). A white powder with mp. 192-194.degree. C., Rf=0.26
(CH.sub.2Cl.sub.2/MeOH=95:5) is obtained.
E.
N-[1-(Cyanomethyl-carbamoyl)-cyclohexyl]-4-(piperazin-1-yl)-benzamide
[0073]
N-[1-(Cyanomethyl-carbamoyl)-cyclohexyl]4-(4-FMOC-piperazin-1-yl)-b-
enzamide (4.4 mmol) is dissolved in DMF (15 ml), piperidine (4.4
mmol) is added and the mixture is stirred at RT for 4 hours. 4
additional drops of piperidine are added and the mixture is stirred
over night. The reaction mixture is poured into water and ethyl
acetate and the suspension is filtered and the filtrate is
acidified with HCl 4N and then extracted with ethyl acetate.
Saturated sodium carbonate solution is added to make the water
phase basic and the mixture is extracted three times with ethyl
acetate. The water phase is saturated with sodium chloride and
extracted three times with ethyl acetate again. The organic
fractions are dried over sodium sulfate and the solvent is
evaporated. The residue is purified by flash chromatography on
silica gel with CH.sub.2Cl.sub.2/MeOH (with 3N NH3)=95:5 as mobile
phase. The product containing fractions are combined and
evaporated. The residue is suspended in diethylether and the solid
filtered of and dried (vacuum). A white powder with mp.
206-210.degree. C., Rf=0.28 (CH.sub.2Cl.sub.2/MeOH (with 3N
NH.sub.3)=9:1) is obtained.
[0074] 1H-NMR (d6-DMSO): 1.15-1.35 (m, 1H); 1.4-1.6 (m, 5H);
1.65-1.8 (m, 2H); 2.05-2.15 (m, 2H); 2.8 (m, 4H); 3.15 (m, 4H); 4.0
(d, 2H), 6.95 (d, 2H); 7.65 (s, 1H); 7.75 (d, 2H), 8.15 (m,
1H).
Example 2
Synthesis of
N-[1-(Cyanomethyl-carbamoyl)-cyclohexyl]-4-(4-methyl-piperazi-
n-1-yl)-benzamide
A. 4-(4-Methyl-piperazin-1-yl)-benzoic acid methyl ester
[0075] 4-Fluorobenzoic acid methyl ester (34 mmol),
1-methyl-piperazine (75 mmol) and potassium carbonate (34 mmol) are
suspended in acetonitrile (30 ml) and stirred under reflux for
three days. After evaporation of the solvent, the residue is
dissolved in water and extracted three times with ethyl acetate.
The extract is dried over sodium sulfate and evaporated. The
residue is purified by flash chromatography on silica gel with
(CH.sub.2Cl.sub.2/MeOH=95:5) as mobile phase. The product
containing fractions are combined and evaporated. The residue is
suspended in diethylether/pentane and the solid filtered of and
dried (vacuum). A pale yellow powder with mp. 117-119.degree. C.,
Rf=0.20 (CH.sub.2Cl.sub.2/MeOH=95:5) is obtained.
B. 4-(4-Methyl-piperazin-1-yl)-benzoic acid hydrochlorid
[0076] 4-(4-Methyl-piperazin-1-yl)-benzoic acid methyl ester (8.5
mmol) is dissolved in 4N HCl (15 ml) and heated under reflux for 8
hours. The mixture is cooled in an ice bath to 0-4.degree. C.,
diluted with 5 ml acetone and the solid material formed is filtered
off, washed with acetone and dried (vacuum). A white powder with
mp. >270.degree. C., Rf=0.11 (CH.sub.2Cl.sub.2/MeOH=9:1) is
obtained.
C.
N-[1-(Cyanomethyl-carbamoyl)-cyclohexyl]-4-(4-methyl-piperazin-1-yl)-be-
nzamide
[0077] 1-Amino-cyclohexanecarboxylic acid cyanomethyl-amide (1.38
mmol) 4-(4-methyl-piperazin-1-yl)-benzoic acid hydrochloride (1.38
mmol), HOBT (1.38 mmol), WSCD (1.38 mmol) and triethylamine (1.38
mmol) are dissolved in DMF (5 ml) and stirred overnight at rt.
After evaporation of the solvent, the residue is dissolved in a
mixture of water and sodium carbonate (to ensure slightly basic
conditions) and extracted three times with ethyl acetate. The
combined extract is dried over sodium sulfate and evaporated. The
residue is suspended in diethylether and the solid filtered of and
dried (vacuum). A pale powder with mp. 218-220.degree. C., Rf=0.19
(CH.sub.2Cl.sub.2/MeOH=9:1) is obtained.
[0078] 1H-NMR (d6-DMSO): 1.15-1.35 (m, 1H); 1.4-1.6 (m, 5H);
1.65-1.8 (m, 2H); 2.05-2.15 (m, 2H); 2.2 (s, 3H); 2.4 (m, 4H); 3.2
(m, 4H); 4.0 (d, 2H), 6.95 (d, 2H); 7.75 (s, 1H); 7.75 (d, 2H),
8.15 (m, 1H).
Example 3
Synthesis of
N-[1-(Cyanomethyl-carbamoyl)-cyclohexyl]-4-(4-ethyl-piperazin-
-1-yl)-benzamide
A. 4-(4-Ethyl-piperazin-1-yl)-benzoic acid methyl ester
[0079] 4-Fluorobenzoic acid methyl ester (53 mmol),
1-ethyl-piperazine (44 mmol) and potassium carbonate (44 mmol) are
suspended in dimethyl-acetamide (50 ml) and stirred under reflux
overnight. After evaporation of the solvent, the residue is
dissolved in water and extracted three times with ethyl acetate.
The extract is dried over sodium sulfate and evaporated. The
residue is suspended in diethylether/pentane and the solid filtered
of and dried (vacuum). A brownish powder with mp. 102-104.degree.
C., Rf=0.22 (CH.sub.2Cl.sub.2/MeOH=95:5) is obtained.
B. 4-(4-Ethyl-piperazin-1-yl)-benzoic acid hydrochlorid
[0080] 4-(4-Ethyl-piperazin-1-yl)-benzoic acid methyl ester (15
mmol) is dissolved in 4N HCl (35 ml) and heated under reflux for 8
hours. The mixture is cooled in an ice bath to 0-4.degree. C. and
the solid material formed is filtered off, washed with acetone and
dried (vacuum). A grey powder with mp. >270.degree. C., Rf=0.08
(CH.sub.2Cl.sub.2/MeOH=9:1) is obtained.
C.
N-[1-(Cyanomethyl-carbamoyl)-cyclohexyl]-4-(4-ethyl-piperazin-1-yl)-ben-
zamide
[0081] 1-Amino-cyclohexanecarboxylic acid cyanomethyl-amide (0.9
mmol) 4-(4-ethyl-piperazin-1-yl)-benzoic acid hydrochloride (0.9
mmol), HOBT (0.9 mmol), WSCD (0.9 mmol) and triethylamine (0.9
mmol) are dissolved in DMF (5 ml) and stirred overnight at rt.
After evaporation of the solvent, the residue is dissolved in a
mixture of water and sodium carbonate (to ensure slightly basic
conditions) and extracted three times with ethyl acetate. The
combined extract is dried over sodium sulfate and evaporated. The
residue is purified by flash chromatography on silica gel with
CH.sub.2Cl.sub.2/MeOH (with 3N NH.sub.3)=93:7 as mobile phase. The
product containing fractions are combined and evaporated. The
residue is suspended in diethylether and the solid filtered of and
dried (vacuum). A white powder is obtained.
[0082] 1H-NMR (d6-DMSO): 1.0 (t, 3H), 1.15-1.35 (m, 1H); 1.4-1.6
(m, 5H); 1.65-1.8 (m, 2H); 2.05-2.15 (m, 2H); 2.35 (q, 2H); 2.45
(m, 4H); 3.2 (m, 4H); 4.0 (d, 2H), 6.95 (d, 2H); 7.65 (s, 1H); 7.75
(d, 2H), 8.15 (m, 1H).
Example 4
Synthesis of
N-[1-(Cyanomethyl-carbamoyl)-cyclohexyl]-4-[4-(1-propyl)-pipe-
razin-1-yl]-benzamide
A. 4-[4-(1-Propyl)-piperazin-1-yl]-benzoic acid methyl ester
[0083] 4-Fluorobenzoic acid methyl ester (165 mmol),
1-(1-propyl)-piperazine dihydro-bromide (138 mmol) and potassium
carbonate (690 mmol) are suspended in dimethyl-acetamide (320 ml)
and stirred under reflux overnight. After evaporation of the
solvent, the residue is dissolved in water and extracted three
times with ethyl acetate. The extract is dried over sodium sulfate
and evaporated. The residue is suspended in diethylether/pentane
and the solid filtered of and dried (vacuum). A brownish powder
with mp. 99-101.degree. C., Rf=0.23 (CH.sub.2Cl.sub.2/MeOH=95:5) is
obtained.
[0084] Cs.sub.2CO.sub.3 may be used in place of K.sub.2CO.sub.3 in
the above procedure.
B. 4-[4-(1-(Propyl)-piperazin-1-yl]-benzoic acid hydrochlorid
[0085] 4-[4-(1-Propyl)-piperazin-1-yl]-benzoic acid methyl ester
(38 mmol) is dissolved in 4N HCl (60 ml) and heated under reflux
for 7 hours. The mixture is cooled in an ice bath to 0-4.degree. C.
and the solid material formed is filtered off, washed with cold
water and dried (vacuum). A pale powder with mp. >270.degree.
C., Rf=0.19 (CH2Cl2/MeOH=9:1) is obtained.
[0086] Alternatively the 4-[4-(1-Propyl)-piperazin-1-yl]-benzoic
acid product may be produced as an internal salt with acetic acid.
For instance, the 4-[4-(1-Propyl)-piperazin-1-yl]-benzoic acid
methyl ester is suspended in water/methanol at 700 .degree. C. and
hydrolysed by addition of 1 equivalent of NaOH; the solution is
clearfiltered and the product precipitated by addition of 1
equivalent of acetic acid, filtered and dried.
C.
N-[1-(Cyanomethyl-carbamoyl)-cyclohexyl]-4-[4-(1-propyl)-piperazin-1-yl-
]-benzamide
[0087] 1-Amino-cyclohexanecarboxylic acid cyanomethyl-amide (22
mmol), 4-[4-(1-propyl)-piperazin-1-yl]-benzoic acid hydrochloride
(22 mmol), HOBT (22 mmol), WSCD (22 mmol) and triethylamine (22
mmol) are dissolved in DMF (50 ml) and stirred overnight at rt.
After evaporation of the solvent, the residue is dissolved in a
mixture of water and sodium carbonate (to ensure slightly basic
conditions) and extracted three times with ethyl acetate. The
combined extract is dried over sodium sulfate and evaporated. The
residue is purified by flash chromatography on silica gel with
(CH.sub.2Cl.sub.2/MeOH=9:1) as mobile phase. The product containing
fractions are combined and evaporated. The residue is suspended in
diethylether and the solid filtered of and dried (vacuum). A white
powder with mp. 216-218.degree. C., Rf=0.34
(CH.sub.2Cl.sub.2/MeOH=9:1) is obtained.
[0088] 1H-NMR (d6-DMSO): 0.85 (t, 3H), 1.2-1.3 (m, 1H); 1.4-1.6 (m,
7H); 1.65-1.8 (m, 2H); 2.05-2.15 (m, 2H); 2.25 (t, 2H); 2.45 (m,
4H); 3.2 (m, 4H); 4.0 (d, 2H), 6.95 (d, 2H); 7.65 (s, 1H); 7.75 (d,
2H), 8.15 (m, 1H).
[0089] In an alternative procedure the acetic acid internal salt of
4-[4-(1-propyl)-piperazin-1-yl]-benzoic acid is treated in
acetonitrile with HOBt, NMM and diisopropylcarbodiimide (DICI), and
after stirring for 1 hr at 40.degree. C. a solution of
1-amino-cyclohexanecarboxylic acid cyanomethyl-amide in
acetonitrile is added. On completion of the reaction, the product
is precipitated by addition of water to the reaction mixture,
filtered and following digestion with ethanol is dried to the end
product.
Example 5
Synthesis of
N-[1-(Cyanomethyl-carbamoyl)-cyclohexyl]-4-(4-isopropyl-piper-
azin-1-yl)-benzamide
A. 4-[4-Isopropyl-piperazin-1-yl]-benzoic acid methyl ester
[0090] Tris-(dibenzylidene-acetone)-dipalladium (0.05 mmol),
(2'-dicyclohexylphosphanyl-biphenyl-2-yl)-dimethyl-amine (0.1 mmol)
and potassium carbonate (4.6 mmol) are suspended in
1,2-dimethoxyethane (10 ml) in an oxygen-free atmosphere (N2).
4-Bromo-benzoic acid methyl ester (3.3 mmol) and
1-isopropyl-piperazine (3.9 mmol) are added and the stirred mixture
is heated under reflux for 28 hours. After cooling the solvent is
evaporated and water is added to the residue, which is then
extracted three times with ethyl acetate. The combined extract is
dried over sodium sulfate and evaporated. The residue is purified
by flash chromatography on silica gel with
(CH.sub.2Cl.sub.2/MeOH=95:5) as mobile phase. The product
containing fractions are combined and evaporated. The residue is
suspended in diethylether/pentane and the solid filtered of and
dried (vacuum). A pale-brown powder with Rf=0.23
(CH.sub.2Cl.sub.2/MeOH=95:5) is obtained.
B. 4-(4-Isopropyl-piperazin-1-yl)-benzoic acid hydrochloride
[0091] 4-(4-Isopropyl-piperazin-1-yl)-benzoic acid methyl ester
(0.9 mmol) is dissolved in 4N HCl (2 ml) and heated under reflux
for 7 hours. The mixture is cooled in an ice bath to 0-4.degree. C.
and acetone is added. The solid material formed is filtered off,
washed with cold acetone and dried (vacuum). A pale-brown powder
with mp. >270.degree. C., Rf=0.08 (CH.sub.2Cl.sub.2/MeOH=9:1) is
obtained.
C.
N-[1-(Cyanomethyl-carbamoyl)-cyclohexyl]-4-(4-isopropyl-piperazin-1-yl)-
-benzamide
[0092] 1-Amino-cyclohexanecarboxylic acid cyanomethyl-amide (0.6
mmol), 4-(4-isopropyl-piperazin-1-yl)-benzoic acid hydrochloride
(0.6 mmol), HOBT (0.6 mmol), WSCD (0.6 mmol) and triethylamine (0.6
mmol) are dissolved in DMF (2 ml) and stirred overnight at rt.
After evaporation of the solvent, the residue is dissolved in a
mixture of water and sodium carbonate (to ensure slightly basic
conditions) and extracted three times with ethyl acetate. The
combined extract is dried over sodium sulfate and evaporated. The
residue is suspended in ethyl acetate/diethylether and the solid
filtered of and dried (vacuum). A white powder with mp.
218-220.degree. C., Rf=0.28 (CH.sub.2Cl.sub.2/MeOH=9:1) is
obtained.
[0093] 1H-NMR (d6-DMSO): 1.0 (d, 6H), 1.2-1.3 (m, 1H); 1.4-1.6 (m,
5H); 1.65-1.8 (m, 2H); 2.05-2.15 (m, 2H); 2.45 (m, 4H); 2.65 (m,
1H); 3.2 (m, 4H); 4.0 (d, 2H), 6.95 (d, 2H); 7.65 (s, 1H); 7.75 (d,
2H), 8.15 (m, 1H).
Example 6
Synthesis of
N-[1-(Cyanomethyl-carbamoyl)-cyclohexyl]-4-(4-benzyl-piperazi-
n-1-yl)-benzamide
A. 4-(4-Benzyl-piperazin-1-yl)-benzoic acid methyl ester
[0094] Tris-(dibenzylidene-acetone)-dipalladium (0.03 mmol),
(2'-dicyclohexylphosphanyl-biphenyl-2-yl)-dimethyl-amine (0.9 mmol)
and NaOtBu (6.5 mmol) are suspended in toluene (20 ml) in an
oxygen-free atmosphere (N2). 4-Bromo-benzoic acid methyl ester
(4.65 mmol) and 1-(benzyl)-piperazine (5.6 mmol) are added and the
stirred mixture is heated under reflux for 4 hours. After cooling,
a mixture of ethylacetate and diethylether is added and the mixture
is filtered. Then the solvent is evaporated and the residue is
suspended in diethylether and the solid filtered of and dried
(vacuum). A pale powder with mp. 105-107.degree. C., Rf=0.67
(CH.sub.2Cl.sub.2/MeOH=95:5) is obtained.
B. 4-(4-Benzyl-piperazin-1-yl)-benzoic acid hydrochloride
[0095] 4-(4-Benzyl-piperazin-1-yl)-benzoic acid methyl ester (0.84
mmol) is dissolved in 4N HCl (2 ml) and heated under reflux for 8
hours. The mixture is cooled in an ice bath to 0-4.degree. C. and
the solid material formed is filtered off, washed with cold acetone
and dried (vacuum). A grey powder with mp. >270.degree. C.,
Rf=0.18 (CH.sub.2Cl.sub.2/MeOH=9- 5:5) is obtained.
C.
N-[1-(Cyanomethyl-carbamoyl)-cyclohexyl]-4-(4-benzyl-piperazin-1-yl)-be-
nzamide
[0096] 1-Amino-cyclohexanecarboxylic acid cyanomethyl-amide (0.84
mmol), 4-[4-(2-propyl)-piperazin-1-yl]-benzoic acid hydrochloride
(0.84 mmol), HOBT (0.84 mmol), WSCD (0.84 mmol) and triethylamine
(0.84 mmol) are dissolved in DMF (2 ml) and stirred overnight at
rt. After evaporation of the solvent, the residue is dissolved in a
mixture of water and sodium carbonate (to ensure slightly basic
conditions) and extracted three times with ethyl acetate. The
combined extract is dried over sodium sulfate and evaporated. The
residue is suspended in methanol and the solid filtered of and
dried (vacuum). A pale powder with mp. 210-212.degree. C., Rf=0.20
(CH.sub.2Cl.sub.2/MeOH=95:5) is obtained.
[0097] 1H-NMR (d6-DMSO): 1.15-1.35 (m, 1H); 1.4-1.6 (m, 5H);
1.65-1.8 (m, 2H); 2.05-2.15 (m, 2H); 2.45 (m, 4H); 3.2 (m, 4H); 3.5
(s, 2H); 4.0 (d, 2H), 6.9 (d, 2H); 7.2-7.4 (m, 5H), 7.65 (s, 1H);
7.75 (d, 2H), 8.15 (m, 1H).
Example 7
Synthesis of
N-[1-(Cyanomethyl-carbamoyl)-cyclohexyl]-4-[4-(2-methoxy-ethy-
l)-piperazin-1-yl]-benzamide
A. 4-(4-Benzyl-piperazin-1-yl)-benzoic acid methyl ester
[0098] 4-Fluorobenzoic acid methyl ester (200 mmol),
1-benzyl-piperazine (300 mmol), and potassium carbonate (300 mmol)
are suspended in acetonitrile (400 ml) and stirred under reflux for
6 days. After evaporation of the solvent, the residue is dissolved
in water and extracted three times with diethylether. The extract
is dried over sodium sulfate and evaporated. The residue is
purified by flash chromatographie on silica gel with
(CH.sub.2Cl.sub.2 first, then CH.sub.2Cl.sub.2/MeOH=15- :1) as
mobile phase. The product containing fractions are combined and
evaporated. The residue is suspended in diethylether/pentane and
the solid filtered of and dried (vacuum). A powder with mp.
105-107.degree. C. is obtained.
B. 4-(Piperazin-1-yl)-benzoic acid methyl ester
[0099] 4-(4-Benzyl-piperazin-1-yl)-benzoic acid methyl ester (19.4
mmol) is dissolved in methanol (150 ml) and Pd/charcoal is added
(0.6 g). The mixture is stirred in a hydrogen atmosphere until
consumption has ceased. The catalyst is filtered off and the
filtrate evaporated. The residue is suspended in
diethylether/pentane and the solid filtered of and dried (vacuum).
A powder with mp. 95-97.degree. C. is obtained.
C. [4-(2-methoxy-ethyl)-piperazin-1-yl]-benzoic acid methyl
ester
[0100] 4-(Piperazin-1-yl)-benzoic acid methyl ester (19 mmol),
2-bromoethylmethylether (21 mmol), and potassium carbonate (22.8
mmol) are suspended in acetonitrile (50 ml) and stirred at
80.degree. C. for 8 hours. After evaporation of the solvent, the
residue is dissolved in water and extracted three times with
CH.sub.2Cl.sub.2. The extract is dried over sodium sulfate and
evaporated. The residue is suspended in diethylether/pentane and
the solid filtered of and dried (vacuum). A powder with mp.
103-105.degree. C. is obtained.
D. [4-(2-methoxy-ethyl)-piperazin-1-yl]-benzoic acid
hydrochloride
[0101] [4-(2-methoxy-ethyl)-piperazin-1-yl]-benzoic acid methyl
ester (17 mmol) is dissolved in 4N HCl (70 ml) and heated under
reflux for 5 hours. After cooling the solvent is evaporated and the
residue is suspended in ethanol and the solid filtered of, washed
with diethylether and dried (vacuum). A powder with mp.
>270.degree. C., Rf=0.35 (CH.sub.2Cl.sub.2/MeOH=9:1) is
obtained.
E.
N-[1-(Cyanomethyl-carbamoyl)-cyclohexyl]-4-[4-(2-methoxy-ethyl)-piperaz-
in-1-yl]-benzamide
[0102] 1-Amino-cyclohexanecarboxylic acid cyanomethyl-amide (1.0
mmol), [4-(2-methoxy-ethyl)-piperazin-1-yl]-benzoic acid
hydrochloride (1.0 mmol), HOBT (1.0 mmol), WSCD (1.0 mmol) and
triethylamine (1.0 mmol) are dissolved in DMF (4 ml) and stirred
overnight at rt. After evaporation of the solvent, the residue is
dissolved in a mixture of water and sodium carbonate (to ensure
slightly basic conditions) and extracted three times with ethyl
acetate. The combined extract is dried over sodium sulfate and
evaporated. The residue is purified by flash chromatography on
silica gel with CH.sub.2Cl.sub.2/MeOH=92.5:7.5 as mobile phase. The
product containing fractions are combined and evaporated. The
residue is suspended in diethylether and the solid filtered of and
dried (vacuum). A pale powder with mp. 166-168.degree. C., Rf=0.37
(CH.sub.2Cl.sub.2/MeOH=9- :1) is obtained.
[0103] 1H-NMR (d6-DMSO): 1.15-1.35 (m, 1H); 1.4-1.6 (m, 5H);
1.65-1.8 (m, 2H); 2.05-2.15 (m, 2H); 2.45 (m, 6H); 3.2 (m, 7H);
3.45 (t, 2H); 4.0 (d, 2H), 6.95 (d, 2H); 7.65 (s, 1H); 7.75 (d,
2H), 8.15 (m, 1H).
Example 8
Synthesis of
N-[1-(Cyanomethyl-carbamoyl)-cyclohexyl]-4-(1-propyl-piperidi-
n-4-yl)-benzamide
A. 1-(4-Phenyl-piperidin-1-yl)-ethanone
[0104] 4-Phenylpiperidine (87 mmol) and pyridine (96 mmol) are
dissolved in dry CH.sub.2Cl.sub.2 (100 ml) and acetylchloride (96
mmol) in CH.sub.2Cl.sub.2 (40 ml) is added dropwise to the stirred
solution at 10.degree. C. The reaction is stirred for 1 hour at rt.
The mixture is extracted three times with water and the water phase
is extracted again with CH.sub.2Cl.sub.2. The combined organic
phases are dried over sodium sulfate and evaporated. A pale brown
oil with Rf=0.13 (ethyl acetate/hexane=1:1) is obtained.
B. 4-Piperidin-4-yl-benzoic acid
[0105] 1-(4-Phenyl-piperidin-1-yl)-ethanone (84 mmol) is dissolved
in CH.sub.2Cl.sub.2 (250 ml) and oxalylchloride (336 mmol) is added
dropwise at -20 to -10.degree. C. After the oxalylchloride addition
aluminium trichloride (260 mmol) is added in portions at
-10.degree. C. The mixture is stirred at -10.degree. C. for 3
hours. The cooling bath is removed and the mixture is stirred at rt
overnight. The mixture is poured on ice/water (600 ml) and
extracted 3 times with CH.sub.2Cl.sub.2. The combined organic
phases are washed with water, dried over sodium sulfate and
evaporated. The residue is dissolved in aqueous sodium hydroxide
solution (2N, 250 ml) and 6N HCl is added at 0.degree. C. to
acidify the solution. The precipitate formed is filtered off and
washed with water. The solid material is suspended in 6N HCl (300
ml) and the mixture is heated for 18 hours under reflux. After
cooling to rt the solvent is removed and the residue is suspended
in ethanol. The solid material is filtered of and dried. A brown
powder with mp. >270.degree. C. is obtained.
C. 4-Piperidin-4-yl-benzoic acid methyl ester
[0106] 4-Piperidin-4-yl-benzoic acid (47 mmol) is dissolved in
methanol (300 ml) and 1 ml of concentrated sulfonic acid is added.
The mixture is heated under reflux overnight. After evaporation of
the solvent, the residue is dissolved in a mixture of water and
sodium carbonate (to ensure basic conditions) and extracted three
times with ethyl acetate. The combined extract is dried over sodium
sulfate and evaporated. A brown powder with Rf=0.18
(CH.sub.2Cl.sub.2/MeOH=9:1) is obtained.
D. 4-(1-Propyl-piperidin-4-yl)-benzoic acid methyl ester
[0107] 4-Piperidin-4-yl-benzoic acid methyl ester (28 mmol),
ethyldiisopropylamine (31 mol) and 1-iodopropane (42 mmol) are
dissolved in 1,2-dimethoxyethane (100 ml) and the mixture is heated
at 70.degree. C. overnight. After evaporation of the solvent, the
residue is dissolved in a mixture of water and sodium carbonate (to
ensure basic conditions) and extracted three times with ethyl
acetate. The combined extract is dried over sodium sulfate and
evaporated. The residue is purified by flash chromatography on
silica gel with CH.sub.2Cl.sub.2/MeOH=9:1 as mobile phase. The
product containing fractions are combined and evaporated. The
residue is suspended in diethylether and the solid filtered of and
dried (vacuum). A pale brown powder with Rf=0.35
(CH.sub.2Cl.sub.2/MeOH=9:1) is obtained.
E. 4-(1-Propyl-piperidin-4-yl)-benzoic acid hydrochloride
[0108] 4-(1-Propyl-piperidin-4-yl)-benzoic acid methyl ester (32
mmol) is dissolved in 4N HCl (45 ml) and heated under reflux for 7
hours. The mixture is cooled in an ice bath to 0-4.degree. C. and
the solid material formed is filtered off, washed with cold acetone
and dried (vacuum). A brown powder with mp. >270.degree. C.,
Rf=0.08 (CH.sub.2Cl.sub.2/MeOH=- 9:1) is obtained.
F.
N-[1-(Cyanomethyl-carbamoyl)-cyclohexyl]-4-(1-propyl-piperidin-4-yl)-be-
nzamide
[0109] 1-Amino-cyclohexanecarboxylic acid cyanomethyl-amide (23
mmol), 4-(1-propyl-piperidin-4-yl)-benzoic acid hydrochloride (23
mmol), HOBT (23 mmol), WSCD (23 mmol) and triethylamine (23 mmol)
are dissolved in DMF (50 ml) and stirred overnight at rt. After
evaporation of the solvent, the residue is dissolved in a mixture
of water and sodium carbonate (to ensure basic conditions) and
extracted three times with ethyl acetate. The combined extract is
dried over sodium sulfate and evaporated. The residue is suspended
in diethylether/pentane and the solid filtered of and dried
(vacuum). A pale powder with mp. 198-200.degree. C., Rf=0.34
(CH.sub.2Cl.sub.2/MeOH=9:1) is obtained.
[0110] 1H-NMR (d6-DMSO): 0.85 (t, 3H); 1.2-1.3 (m, 1H); 1.4-1.6 (m,
7H); 1.6-1.8 (m, 6H); 1.9-2.0 (m, 2H); 2.05-2.15 (m, 2H); 2.25 (t,
2H); 2.55 (m, 1H); 2.95 (d, 2H); 4.0 (d, 2H), 7.35 (d, 2H); 7.8 (d,
2H), 7.9 (s, 1H); 8.15 (m, 1H).
Example 9
Synthesis of
N-[1-(Cyanomethyl-carbamoyl)-cyclohexyl]-4-[1-(2-methoxy-ethy-
l)-piperidin-4-yl]-benzamide
A. 4-Carboxybenzeneboronic acid methyl ester
[0111] 4-Carboxybenzeneboronic acid (300 mmol) is dissolved in
methanol (400 ml) and 1.5 ml concentrated HCl is added to the
stirred solution. The reaction is heated under reflux for 30 hours.
The solvent is evaporated, the remaining residue is suspended in
diethylether and the solid filtered of and dried (vacuum). A pale
powder with mp. 201-205.degree. C., Rf=0.28
(CH.sub.2Cl.sub.2/MeOH=95:5) is obtained. This powder is a mixture
of 4-carboxybenzeneboronic acid methyl ester and the dimeric
anhydride of 4-carboxybenzeneboronic acid methyl ester and is used
without further purification.
B. 4-Pyridin-4-yl-benzoic acid methyl ester
[0112] 4-Carboxybenzeneboronic acid methyl ester (248 mmol) from A,
4-bromopyridine (248 mmol), tetrakis-(triphenylphosphin)-palladium
(2.5 mmol) and potassium carbonate (744 mmol) are suspended in
1,2-dimethoxyethane (1100 ml). The stirred mixture is heated under
reflux for 8 hours. After cooling the solvent is evaporated and
water is added to the residue which is then extracted three times
with ethyl acetate. The combined extract is dried over sodium
sulfate and evaporated. The residue is suspended in diethylether
and the solid filtered of and dried (vacuum). A pale-brown powder
with mp. 99-101.degree. C., Rf=0.39 (CH.sub.2Cl.sub.2/MeOH=95:5) is
obtained.
C. 4-(4-Methoxycarbonyl-phenyl)-1-(2-methoxy-ethyl)-pyridinium:
bromide
[0113] 4-Pyridin-4-yl-benzoic acid methyl ester (70 mmol) and
2-bromoethyl-methylether (28 ml) are heated for 1 hour to
110.degree. C. After cooling the reaction mixture is suspended in
acetone and the solid filtered of and dried (vacuum). A pale-brown
powder with mp. 170-171.degree. C., Rf=0.22
(CH.sub.2Cl.sub.2/MeOH=9:1) is obtained.
D. 4-[1-(2-Methoxy-ethyl)-piperidin-4-yl]-benzoic acid methyl
ester
[0114] 4-(4-Methoxycarbonyl-phenyl)-1-(2-methoxy-ethyl)-pyridinium;
bromide (67 mmol) is suspended in methanol (250 ml) and platinoxide
(1.2 g) is added. The mixture is stirred in a hydrogen atmosphere
at normal pressure until consumption has ceased. The catalyst is
filtered off and the filtrate evaporated. The residue is dissolved
in CH.sub.2Cl.sub.2 and extracted with aqueous sodium carbonate
solution. The organic phase is dried over sodium sulfate and
evaporated. The residue is purified by flash chromatography on
silica gel with CH.sub.2Cl.sub.2/MeOH=9:1 as mobile phase. The
product containing fractions are combined and evaporated. A pale
yellow oil with Rf=0.22 (CH.sub.2Cl.sub.2/MeOH=95:5) is
obtained.
E. 4-[1-(2-Methoxy-ethyl)-piperidin-4-yl]-benzoic acid
hydrochloride
[0115] 4-[1-(2-Methoxy-ethyl)-piperidin-4-yl]-benzoic acid methyl
ester (47 mmol) is dissolved in 4N HCl (80 ml) and heated under
reflux for 12 hours. After cooling the solvent is evaporated and
the residue is suspended in acetone and the solid filtered of,
washed with acetone and dried (vacuum). A white powder with mp.
>270.degree. C. is obtained.
F.
N-[1-(Cyanomethyl-carbamoyl)-cyclohexyl]-4-[1-(2-methoxy-ethyl)-piperid-
in-4-yl]-benzamide
[0116] 1-Amino-cyclohexanecarboxylic acid cyanomethyl-amide (107
mmol), 4-[1-(2-methoxy-ethyl)-piperidin-4-yl]-benzoic acid
hydrochloride (107 mmol), HOBT (107 mmol), WSCD (107 mmol) and
triethylamine (107 mmol) are dissolved in DMF (250 ml) and stirred
overnight at rt. After evaporation of the solvent, the residue is
dissolved in a mixture of water and sodium carbonate (to ensure
slightly basic conditions) and extracted three times with ethyl
acetate. The combined extract is dried over sodium sulfate and
evaporated. The residue is purified by flash chromatography on
silica gel with CH.sub.2Cl.sub.2/MeOH (with 2N NH.sub.3)=9:1 as
mobile phase. The product containing fractions are combined and
evaporated. The residue is suspended in diethylether/ethyl acetate
and the solid filtered of and dried (vacuum). A pale powder with
mp. 160-162.degree. C., Rf=0.42 (CH.sub.2Cl.sub.2/MeOH (with 3N
NH.sub.3)=9:1) is obtained.
[0117] 1H-NMR (d6-DMSO): 1.2-1.3 (m, 1H); 1.4-1.6 (m, 5H); 1.6-1.8
(m, 6H); 2.0-2.2 (m, 4H); 2.45 (m, 2H); 2.55 (m, 1H); 2.95 (br. d,
2H); 3.2 (s, 3H); 3.4 (dd, 2H); 4.0 (d, 2H); 7.35 (d, 2H); 7.8 (d,
2H); 7.9 (s, 1H); 8.15 (m, 1H).
Example 10
N-[1-(Cyanomethyl-carbamoyl)-cyclohexyl]-4-(1-isopropyl-piperidin-4-yl)-be-
nzamide
A. 1-Isopropyl-4-(4-methoxycarbonyl-phenyl)-pyridiniumm;
bromide
[0118] 4-Pyridin-4-yl-benzoic acid methyl ester (2.3 mmol) and
2-iodopropane (1.0 ml) are heated for 24 hours to 90.degree. C.
After cooling the solvent is evaporated and the residue is
suspended in acetone and the solid filtered of and dried (vacuum).
A pale-yellow powder with mp. 187-189.degree. C., Rf=0.27
(CH.sub.2Cl.sub.2/MeOH=9:1) is obtained.
B. 4-(1-Isopropyl-piperidin-4-yl)-benzoic acid methyl ester
hydroiodide
[0119] 1-Isopropyl-4-(4-methoxycarbonyl-phenyl)-pyridinium; bromide
(1.9 mmol) is suspended in methanol (10 ml) and platinoxide (80 mg)
is added. The mixture is stirred in a hydrogen atmosphere at normal
pressure until consumption has ceased. The catalyst is filtered off
and the filtrate evaporated. The residue is suspended in
diethylether/pentane and the solid filtered of and dried (vacuum).
A pale powder with mp. 219-224.degree. C., Rf=0.41
(CH.sub.2Cl.sub.2/MeOH=9:1) is obtained.
C. 4-(1-Isopropyl-piperidin-4-yl)-benzoic acid hydrochloride
[0120] 4-(1-Isopropyl-piperidin-4-yl)-benzoic acid methyl ester
hydroiodide (1.7 mmol) is dissolved in 4N HCl (5 ml) and heated
under reflux for 10 hours. After cooling the solvent is evaporated
and the residue is suspended in acetone and the solid filtered of,
washed with acetone and dried (vacuum). A grey-brown powder with
mp. >270.degree. C. is obtained.
D.
N-[1-(Cyanomethyl-carbamoyl)-cyclohexyl-4-(1-isopropyl-piperidin-4-yl)--
benzamide
[0121] 1-Amino-cyclohexanecarboxylic acid cyanomethyl-amide (0.95
mmol), 4-(1-isopropyl-piperidin-4-yl)-benzoic acid hydrochloride
(0.95 mmol), HOBT (0.95 mmol), WSCD (0.95 mmol) and triethylamine
(0.95 mmol) are dissolved in DMF (5 ml) and stirred overnight at
rt. After evaporation of the solvent, the residue is dissolved in a
mixture of water and sodium carbonate (to ensure basic conditions)
and extracted three times with ethyl acetate. The combined extract
is dried over sodium sulfate and evaporated. The residue is
suspended in diethylether and the solid filtered of and dried
(vacuum). A white powder with mp. 214-216.degree. C., Rf=0.38
(CH.sub.2Cl.sub.2/MeOH (with 3N NH.sub.3)=9:1) is obtained.
[0122] 1H-NMR (d6-DMSO): 0.95 (d, 6H); 1.2-1.3 (m, 1H); 1.4-1.8 (m,
11H); 2.05-2.25 (m, 4H); 2.55 (m, 1H); 2.7 (m, 1H); 2.85 (d, 2H);
4.0 (d, 2H), 7.35 (d, 2H); 7.8 (d, 2H), 7.9 (s, 1H); 8.15 (m,
1H).
Example 11
N-[1-(Cyanomethyl-carbamoyl)-cyclohexyl]-4-(1-cyclopentyl-piperidin-4-yl)--
benzamide
A. 1-Cyclopentyl -4-(4-methoxycarbonyl-phenyl)-pyridinium;
bromide
[0123] 4-Pyridin4-yl-benzoic acid methyl ester (2.35 mmol) and
1-iodocyclopentane (1.0 ml) are heated for 4 hours to 110.degree.
C. 1-Iodocyclopentane (0.5 ml) are added and the mixture is heated
for another 4 hours to 120.degree. C. After cooling the solvent is
evaporated and the residue is suspended in acetone and the solid
filtered of and dried (vacuum). The solid residue is purified by
flash chromatography on silica gel with CH.sub.2Cl.sub.2/MeOH=9:1
as mobile phase. The product containing fractions are combined and
evaporated. The residue is suspended in diethylether and the solid
filtered of and dried (vacuum). A yellow powder with mp.
183-185.degree. C., Rf=0.35 (CH.sub.2Cl.sub.2/MeOH=9:1) is
obtained.
B. 4-(1-Cyclopentyl-piperidin-4-yl)-benzoic acid methyl ester
hydro-iodide
[0124] 1-Cyclopentyl-4-(4-methoxycarbonyl-phenyl)-pyridinium;
bromide (1.27 mmol) is suspended in methanol (8 ml) and platinoxide
(50 mg) is added. The mixture is stirred in a hydrogen atmosphere
at normal pressure until consumption has ceased. The catalyst is
filtered off and the filtrate evaporated. The residue is suspended
in diethylether/pentane and the solid filtered of and dried
(vacuum). A pale powder with mp. 204-210.degree. C., Rf=0.27
(CH.sub.2Cl.sub.2/MeOH=95:5) is obtained.
C. 4-(1-Cyclopentyl-piperidin-4-yl)-benzoic acid hydrochloride
[0125] 4-(1-Cyclopentyl-piperidin-4-yl)-benzoic acid methyl ester
hydroiodide (1.06 mmol) is dissolved in 4N HCl (5 ml) and heated
under reflux for 10 hours. After cooling the solvent is evaporated
and the residue is suspended in acetone and the solid filtered of,
washed with acetone and dried (vacuum). A grey-brown powder with
mp. >270.degree. C. is obtained.
D.
N-[1-(Cyanomethyl-carbamoyl)-cyclohexyl]-4-(1-cyclopentyl-piperidin-4-y-
l)-benzamide
[0126] 1-Amino-cyclohexanecarboxylic acid cyanomethyl-amide (0.74
mmol), 4-(1-cyclopentyl-piperidin-4-yl)-benzoic acid hydrochloride
(0.74 mmol), HOBT (0.74 mmol), WSCD (0.74 mmol) and triethylamine
(0.74 mmol) are dissolved in DMF (5 ml) and stirred overnight at
rt. After evaporation of the solvent, the residue is dissolved in a
mixture of water and sodium carbonate (to ensure basic conditions)
and extracted three times with ethyl acetate. The combined extract
is dried over sodium sulfate and evaporated. The residue is
suspended in diethylether and the solid filtered of and dried
(vacuum). A white powder with mp. 233-234.degree. C., Rf=0.34
(CH.sub.2Cl.sub.2/MeOH (with 3N NH.sub.3)=9:1) is obtained.
[0127] 1H-NMR (d6-DMSO): 1.2-1.85 (m, 20H); 1.9-2.15 (m, 4H);
2.4-2.6 (m, 2H); 3.05 (d, 2H); 4.0 (d, 2H), 7.35 (d, 2H); 7.8 (d,
2H), 7.9 (s, 1H); 8.15 (m, 1H).
Example 12
N-[1-(Cyanomethyl-carbamoyl)-cyclohexyl]-4-(1-methyl-piperidin-4-yl)-benza-
mide
A. 4-Phenyl-1-methyl-piperidine
[0128] 4-Phenylpiperidine (12.4 mmol), paraformaldehyde (24.8 mmol)
and tetraisopropoxy-titanium (12.4 mmol) are suspended in
1,2-dimethoxyethane (20 ml) and warmed to 60.degree. C. for 30
minutes and stirred at rt for one additional hour. Sodium
borohydride (12.4 mmol) is added in portions and the mixture is
stirred at rt for 2 hours and at 60.degree. C. for additional 3
hours. After cooling the solvent is evaporated and the residue is
dissolved in a mixture of aqueous ammonia (60 ml) and ethyl acetate
and filtered carefully. The mixture is extracted three times with
ethyl acetate and the combined organic phases are dried over sodium
sulfate and evaporated. A pale brown oil is obtained.
B. 4-(1-Methyl-piperidin-4-yl)-benzoic acid methyl ester
[0129] 4-Phenyl-1-methyl-piperidine (9.9 mmol) is dissolved in
CH.sub.2Cl.sub.2 (60 ml) and oxalylchloride (39.6 mmol) is added
dropwise at -20 to -10.degree. C. After the oxalylchloride addition
aluminium trichloride (260 mmol) is added in portions at
-10.degree. C. The mixture is stirred at -10.degree. C. for 1.5
hours. Then the cooling bath is removed and the mixture is stirred
at rt for another 2 hours. The mixture is cooled again to
-0.degree. C. and methanol (30 ml) is added dropwise. After
completion of the methanol addition the cooling bath is removed and
the mixture is stirred at rt overnight. The reaction mixture is
poured into a mixture of aqueous sodium carbonate (to ensure basic
conditions) and ethyl acetate and the suspension is filtered
carefully. The filtrate is extracted three times with ethyl acetate
and the combined extract is dried over sodium sulfate and
evaporated. The residue is purified by flash chromatography on
silica gel with CH.sub.2Cl.sub.2/MeOH=9:1 as mobile phase. The
product containing fractions are combined and evaporated. A pale
yellow oil with Rf=0.29 (CH.sub.2Cl.sub.2/MeOH=9:1) is
obtained.
C. 4-(1-Methyl-piperidin-4-yl)-benzoic acid hydrochloride
[0130] 4-(1-Methyl-piperidin-4-yl)-benzoic acid methyl ester (4.55
mmol) is dissolved in 4N HCl (10 ml) and heated under reflux for 8
hours. After cooling the solvent is evaporated and the residue is
suspended in acetone and the solid filtered of, washed with acetone
and dried (vacuum). A pale-brown powder with mp. >270.degree. C.
is obtained.
D.
N-[1-(Cyanomethyl-carbamoyl)-cyclohexyl]-4-(1-methyl-piperidin-4-yl)-be-
nzamide
[0131] 1-Amino-cyclohexanecarboxylic acid cyanomethyl-amide (0.98
mmol), 4-(1-methyl-piperidin-4-yl)-benzoic acid hydrochloride (0.98
mmol), HOBT (0.98 mmol), WSCD (0.98 mmol) and triethylamine (0.98
mmol) are dissolved in DMF (5 ml) and stirred overnight at rt.
After evaporation of the solvent, the residue is dissolved in a
mixture of water and sodium carbonate (to ensure basic conditions)
and extracted three times with ethyl acetate. The combined extract
is dried over sodium sulfate and evaporated. The residue is
suspended in diethylether/pentane and the solid filtered of and
dried (vacuum). A white powder with mp. 215-217.degree. C., Rf=0.32
(CH.sub.2Cl.sub.2/MeOH (with 3N NH.sub.3)=9:1) is obtained.
[0132] 1H-NMR (d6-DMSO): 1.2-1.3 (m, 1H); 1.4-1.8 (m, 11H);
1.85-2.0 (m, 2H); 2.05 -2.2 (m, 5H); 2.55 (m, 1H); 2.95 (d, 2H);
4.0 (d, 2H), 7.35 (d, 2H); 7.8 (d, 2H), 7.9 (s, 1H); 8.15 (m,
1H).
[0133] Similarly
N-[1-(cyanomethyl-carbamoyl)-cyclohexyl]4-(piperidin-4-yl-
)-benzamide is obtained substantially as described above in Example
12; for instance by omitting Step A and starting the synthesis
procedure at step B, using 4-phenylpiperidine as the starting
material.
* * * * *