5-[3-[PIPERAZIN-1-YL]-3-OXO-PROPYL]-IMIDAZOLIDINE-2,4-DIONE DERIVATIVES AS ADAMTS 4 AND 5 INHIBITORS FOR TREATING E.G. OSTEOARTHRITIS
20190315719 · 2019-10-17
Inventors
- Frédéric Gilbert LABÉGUÈRE (Frouzins, FR)
- Rama Heng (Paris, FR)
- Frédéric André DE CEUNINCK (Paris, FR)
- Luke Jonathan Alvey (Romainville, FR)
- David Amantini (Romainville, FR)
- Franck Laurent Brebion (Romainville, FR)
- Pierre Marc Marie Joseph Deprez (Romainville, FR)
- Romain Luc Marie Gosmini (Romainville, FR)
- Hélène Marie JARY (Romainville, FR)
- Christophe Peixoto (Romainville, FR)
- Iuliana Ecaterina Pop-Botez (Houilles, FR)
- Marie Laurence Claire Varin (Romainville, FR)
Cpc classification
C07D403/06
CHEMISTRY; METALLURGY
C07D401/04
CHEMISTRY; METALLURGY
International classification
C07D403/06
CHEMISTRY; METALLURGY
Abstract
The present invention discloses compounds according to Formula I:
##STR00001##
Wherein R.sup.1, R.sup.2, R.sup.3a, R.sup.3b, R.sup.6a, R.sup.6b, the subscript n and Cy are as defined herein.
The present invention relates to compounds inhibiting ADAMTS, methods for their production, pharmaceutical compositions comprising the same, and methods of treatment using the same, for the prophylaxis and/or treatment of inflammatory diseases, and/or diseases involving degradation of cartilage and/or disruption of cartilage homeostasis by administering the compound of the invention.
Claims
1. A compound according to Formula I: ##STR00381## Wherein R.sup.1 is: H, C.sub.1-4 alkyl optionally substituted with one or more independently selected R.sup.4 groups, C.sub.3-7 monocyclic cycloalkyl optionally substituted with one or more independently selected R.sup.4 groups, 4-7 membered monocyclic heterocycloalkyl comprising 1 to 2 heteroatoms independently selected from N, O, and S, optionally substituted with one or more independently selected C.sub.1-4 alkyl, C(O)C.sub.1-4 alkyl, or C(O)OC.sub.1-4 alkyl, phenyl optionally substituted with one or more independently selected R.sup.5 groups, phenyl fused to a 5-6 membered monocyclic heterocycloalkyl comprising 1, 2 or 3 heteroatoms independently selected from N, O, and S, which heterocycloalkyl is optionally substituted with one or more O, 5-6 membered monocyclic heteroaryl comprising 1 or 2 heteroatoms independently selected from N, O, and S, optionally substituted with one or more independently selected R.sup.5 groups; R.sup.2 is independently selected from: H, OH, C.sub.1-4 alkoxy, and C.sub.1-4 alkyl optionally substituted with one OH, CN, C.sub.1-4 alkoxy optionally substituted with one phenyl, and 5-6 membered monocyclic heteroaryl comprising 1 or 2 heteroatoms independently selected from N, O, and S, optionally substituted with one or more independently selected C.sub.1-4 alkyl; each R.sup.3a, and R.sup.3b is independently selected from H, and C.sub.1-4 alkyl optionally substituted with one or more halo; Cy is 6-10 membered monocyclic or fused bicyclic aryl, 5-10 membered monocyclic or fused bicyclic heteroaryl comprising 1, 2 or 3 heteroatoms independently selected from N, O, and S; R.sup.4 is halo, OH, CN, C.sub.1-4 alkyl, C.sub.1-4 alkoxy optionally substituted with one C.sub.1-4 alkoxy, or phenyl, C.sub.1-4 thioalkoxy, 4-7-membered monocyclic heterocycloalkyl comprising one or more heteroatoms independently selected from N, S, and O, optionally substituted with one or more halo, or C(O)OC.sub.1-4 alkyl, phenyl, S(O).sub.2C.sub.1-4 alkyl C(O)OR.sup.7a C(O)NR.sup.7bR.sup.7c NHC(O)OR.sup.7d NHC(O)R.sup.7e NR.sup.8aR.sup.8b; each R.sup.5 is halo, OH, CN, C.sub.1-4 alkyl optionally substituted with one or more independently selected halo, NR.sup.9aR.sup.9b, or C(O)NR.sup.9cR.sup.9d, C.sub.1-4 alkoxy optionally substituted with NR.sup.9eR.sup.9f, or S(O).sub.2C.sub.1-4 alkyl; each R.sup.6a is C.sub.2-4 alkyl optionally substituted with one or more halo, C.sub.1-4 alkoxy optionally substituted with one or more halo, the subscript n is 0, 1, 2 or 3 each R.sup.6b is independently selected from halo, CN, NO.sub.2, C.sub.1-4 alkyl, C.sub.1-4 alkoxy 5-10 membered monocyclic or fused bicyclic heteroaryl comprising 1, 2 or 3 heteroatoms independently selected from N, O, and S, optionally substituted with one or more independently selected halo, C.sub.1-4 alkyl, or C.sub.1-4 alkoxy, and NR.sup.9gR.sup.9h; each R.sup.7a, R.sup.7b, R.sup.7c, R.sup.7d, or R.sup.7e, is H, or C.sub.1-4 alkyl optionally substituted with OH or C.sub.1-4 alkoxy; each R.sup.8a, or R.sup.8b is independently selected from H, and C.sub.1-4 alkyl optionally substituted with OH, C.sub.1-4 alkoxy, or phenyl; each R.sup.9a, R.sup.9b, R.sup.9c, R.sup.9d, R.sup.9e, R.sup.9f, R.sup.9g, R.sup.9h is independently selected from H, and C.sub.1-4 alkyl; or a pharmaceutically acceptable salt, or a solvate, or a pharmaceutically acceptable salt of a solvate thereof; or a biologically active metabolite thereof; provided that when R.sup.1, R.sup.2, R.sup.3a and R.sup.3b are H, Cy is phenyl and R.sup.6a is OCH.sub.3, then the subscript n is not 0, and the compound is not (5R)-5-[3-[(3S)-4-(4-chloro-3-methoxy-5-methyl-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione.
2. A compound of pharmaceutically acceptable salt thereof according to claim 1, wherein R.sup.1 is H, C.sub.1-4 alkyl substituted with one C.sub.1-4 alkoxy, or C.sub.3-7 monocyclic cycloalkyl.
3. A compound of pharmaceutically acceptable salt thereof according to claim 1, wherein R.sup.2 is H, or C.sub.1-4 alkyl.
4. A compound of pharmaceutically acceptable salt thereof according to claim 1, wherein R.sup.3a is H.
5. A compound of pharmaceutically acceptable salt thereof according to claim 1, wherein R.sup.3b is C.sub.1-4 alkyl.
6. A compound of pharmaceutically acceptable salt thereof according to claim 1, wherein the compound is according to Formula Va, Vb, Vc, or Vd: ##STR00382##
7. A compound of pharmaceutically acceptable salt thereof according to claim 1, wherein Cy is phenyl.
8. A compound of pharmaceutically acceptable salt thereof according to claim 1, wherein R.sup.6a is CH.sub.2CH.sub.3, CH(CH.sub.3).sub.2, OCH.sub.3, or OCH.sub.2CH.sub.3, each of which is optionally substituted with one or more F.
9. A compound of pharmaceutically acceptable salt thereof according to claim 1, wherein the subscript n is 0.
10. A compound of pharmaceutically acceptable salt thereof according to claim 1, wherein the subscript n is 1 or 2.
11. A compound of pharmaceutically acceptable salt thereof according to claim 10, wherein R.sup.6b is halo, C.sub.1-4 alkyl, or C.sub.1-4 alkoxy.
12. A compound or pharmaceutically acceptable salt thereof according to claim 1, wherein the compound is: 5-[3-[4-(3-methoxyphenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, 5-cyclopropyl-5-[3-[4-(3-methoxyphenyl)piperazin-1-yl]-3-oxo-propyl]imidazolidine-2,4-dione, 5-methyl-5-[3-oxo-3-[4-[4-(trifluoromethoxy)phenyl]piperazin-1-yl]propyl]imidazolidine-2,4-dione, 5-methyl-5-[3-oxo-3-[4-[2-(trifluoromethoxy)phenyl]piperazin-1-yl]propyl]imidazolidine-2,4-dione, 5-[3-[4-(4-methoxyphenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, 5-[3-[4-(3-methoxyphenyl)piperazin-1-yl]-3-oxo-propyl]-5-(3-pyridyl)imidazolidine-2,4-dione, 5-[3-[4-(2-methoxyphenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, 5-[3-[4-(3-methoxyphenyl)piperazin-1-yl]-3-oxo-propyl]-5-(2-pyridyl)imidazolidine-2,4-dione, 5-[3-[4-(4-fluoro-3-methoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, 5-[3-[4-(4-chloro-3-methoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, 5-[3-[4-(3,5-dimethoxyphenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, 5-[3-[4-(3-fluoro-5-methoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, 5-[3-[4-(3-chloro-5-methoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, 5-[3-[4-(3,4-difluoro-5-methoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, 5-cyclopropyl-5-[3-[4-(3-fluoro-5-methoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]imidazolidine-2,4-dione, 5-[3-[4-(3-ethoxyphenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, 5-cyclopropyl-5-[3-[4-(4-fluoro-3-methoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]imidazolidine-2,4-dione, 5-cyclopropyl-5-[3-[4-(3,4-difluoro-5-methoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]imidazolidine-2,4-dione, (R)-5-[3-[4-(3-fluoro-5-methoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, (R)-5-[3-[4-(3,4-difluoro-5-methoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, 5-cyclopropyl-5-[3-[(3S)-4-(3,4-difluoro-5-methoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]imidazolidine-2,4-dione, 5-[3-[(3 S)-4-(3,4-difluoro-5-methoxy-phenyl) -3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, 5-[3-[4-(3-fluoro-5-methoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]-5-(3-pyridyl)imidazolidine-2,4-dione, 5-[3-[(3 S)-4-(4-chloro-3-methoxy-phenyl) -3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, 5-[3-[(3 S)-4- (3,5-dimethoxyphenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, (5S)-5-cyclopropyl-5-[(2S)-3-[4-(3-fluoro-5-methoxy-phenyl)piperazin-1-yl]-2-methyl-3-oxo-propyl]imidazolidine-2,4-dione, (5R) -5-[(2S) -3-[4-(3-fluoro-5-methoxy-phenyl) piperazin-1-yl]-2-methyl-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, 5-[3-[4-(3,4-difluoro-5-methoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]-5-(3-pyridyl)imidazolidine-2,4-dione, 5-cyclopropyl-5-[(2S)-3-[(3S) -4-(3,4-difluoro-5-methoxy-phenyl)-3-methyl-piperazin-1-yl]-2-methyl-3-oxo-propyl]imidazolidine-2,4-dione, 5-[3-[(3S)-4-(4-chloro-3,5-dimethoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, 5-[3-[4-(4-chloro-3,5-dimethoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, 5-[3-[4-(5-methoxy-2-methyl-phenyl) piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, 5-[3-[4-(3-methoxy-2-methyl-phenyl) piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, 5-[3-[4-(4-chloro-5-methoxy-2-methyl-phenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, 5-[3-[(3S)-4-(5-methoxy-2-methyl-phenyl) -3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, 5-[3-[(3S)-4-(4-chloro-5-methoxy-2-methyl-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, (5S)-5-cyclopropyl-5-[3-[(3S)-4-(3,4-difluoro-5-methoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]imidazolidine-2,4-dione, 5-[3-[4-(4-chloro-3-fluoro-5-methoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, 5-[3-[(3S)-4-(4-chloro-3-fluoro-5-methoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, 5-[3-[4-(4-chloro-3,5-dimethoxy-phenyl) piperazin-1-yl]-2-methyl-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, 5-[3-[4-(4-chloro-3-methoxy-phenyl)-3-methyl-piperazin-1-yl]-2-methyl-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, 5-[3-[(3S)-4-(3,4-difluoro-5-methoxy-phenyl)-3-methyl-piperazin-1-yl]-2-methyl-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, 5-[3-[4-(3,4-difluoro-5-methoxy-phenyl)piperazin-1-yl]-2-methyl-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, 5-[3-[(3S)-4-(4-chloro-3,5-dimethoxy-phenyl)-3-methyl-piperazin-1-yl]-2-methyl-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, 5-[3-[(3S)-4-(4-chloro-5-methoxy-2-methyl-phenyl)-3-methyl-piperazin-1-yl]-2-methyl-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, 5-[3-[(3S)-4-(3,4-difluoro-5-methoxy-phenyl)-3-methyl-piperazin-1-yl]-2-methyl-3-oxo-propyl]-5-(methoxymethyl) imidazolidine-2,4-dione, 5-[3-[(3 S)-4-(4-chloro-3-methoxy-phenyl) -3-methyl-piperazin-1-yl]-2-methyl-3-oxo-propyl]-5-(methoxymethyl) imidazolidine-2,4-dione, 5-[3-[4-(3,4-dichloro-5-methoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, 5-[3-[(3S)-4-(3,4-difluoro-5-methoxy-phenyl)-3-methyl-piperazin-1-yl]-2-methyl-3-oxo-propyl]-5-(2-pyridyl)imidazolidine-2,4-dione, 5-[3-[4-(3-chloro-4-fluoro-5-methoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, 5-[3-[4-(3,4-difluoro-5-methoxy-phenyl)piperazin-1-yl]-2-methyl-3-oxo-propyl]imidazolidine-2,4-dione, 5-[3-[4-(3,4-difluoro-5-methoxy-phenyl)piperazin-1-yl]-2-methyl-3-oxo-propyl]-5-[(3,3-difluoropyrrolidin-1-yl)methyl]imidazolidine-2,4-dione, 5-[3-[(3S)-4-(3-chloro-4-fluoro-5-methoxy-phenyl) -3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, (5S)-5-[(2S)-3-[4-(3,4-difluoro-5-methoxy-phenyl)piperazin-1-yl]-2-methyl-3-oxo-propyl]-5-(2-pyridyl)imidazolidine-2,4-dione, (5R)-5-[3-[(3S)-4-(4-chloro-3-methoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, (5R)-5-[(2R)-3-[4-(3,4-difluoro-5-methoxy-phenyl)piperazin-1-yl]-2-(hydroxymethyl)-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, (5R)-5-[(2S) -3-[4-(3,4-difluoro-5-methoxy-phenyl) piperazin-1-yl]-2-methyl-3-oxo-propyl]-5-ethyl-imidazolidine-2,4-dione, 5-[3-[4-(4-chloro-2-methoxy-5-methyl-phenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, (5S)-5-[(2S)-3-[4-(3,4-difluoro-5-methoxy-phenyl)piperazin- -yl]-2-methyl-3-oxo-propyl]-5-(methoxymethyl) imidazolidine-2,4-dione, 5-cyclopropyl-5-[3-[(3S)-4-(5-methoxy-3-pyridyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]imidazolidine-2,4-dione, 5-[3-[(3S)-4-(3,4-difluoro-5-methoxy-phenyl) -3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-(6-methyl-2-pyridyl) imidazolidine-2,4-dione, (5S)-5-[3-[(3S)-4-(3-chloro-4-fluoro-5-methoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-cyclopropyl-imidazolidine-2,4-dione, (5R)-5-[3-[(3S)-4-(3-chloro-4-fluoro-5-methoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, 5-cyclopropyl-5-[3-[(3S) -4-(5-methoxy-3-pyridyl)-3-methyl-piperazin-1-yl]-2-methyl-3-oxo-propyl]imidazolidine-2,4-dione, (5S)-5-[3-[(3S)-4-(4-chloro-3-fluoro-5-methoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-cyclopropyl-imidazolidine-2,4-dione, (5R)-5-[3-[(3S)-4-(4-fluoro-5-methoxy-2-methyl-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, (5R) -5-[3-[4-(4-fluoro-5-methoxy-2-methyl-phenyl) piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, (5R)-5-[3-[4-(3-chloro-2-methoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, (5R)-5-[3-[(3S)-4-(3-chloro-2-methoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, (5R)-5-[3-[4-(3-ethylphenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, (5R)-5-[3-[4-(4-chloro-3-ethyl-phenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, (5R)-5-[3-[4-(3,5-diethylphenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, (5R)-5-[3-[4-(4-chloro-2-methoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, (5R)-5-[3-[4-(4-fluoro-3,5-dimethoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, (5R)-5-[3-[(3S)-4-(4-chloro-3-ethyl-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, (5R)-5-[3-[(3S)-4-(4-chloro-2-methoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, (5R)-5-[3-[4-(5-chloro-2-methoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, (5R)-5-[3-[4-(4-fluoro-2-methoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, (5R)-5-[3-[4-(4-chloro-3-ethoxy-5-methoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, (5R)-5-[3-[(3S)-4-(4-chloro-3-ethoxy-5-methoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, (5R)-5-[3-[(3S)-4-(4-fluoro-2-methoxy-phenyl) -3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, (5R)-5-[3-[4-(3-fluoro-2-methoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, (5R)-5-[3-[4-(5-fluoro-2-methoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, (5R)-5-[3-[(3S)-4-(3-fluoro-2-methoxy-phenyl) -3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, (5R)-5-[3-[(3S)-4-(4-chloro-3-isopropyl-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, (5R)-5-[3-[(3S)-4-[4-chloro-3-(trifluoromethoxy)phenyl]-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, (5R) -5-[3-[4-(4-chloro-3-isopropyl-phenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, (5R)-5-[3-[4-[4-chloro-3-(trifluoromethoxy)phenyl]piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, (5R)-5-[3-[(3S)-4-(4-fluoro-3,5-dimethoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, (5R)-5-[3-[(3S)-4-(3,4-dichloro-5-methoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, (5R)-5-[3-[(3S)-4-(5-chloro-2-methoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, (5R)-5-[3-[(3S)-4-(4,5-dichloro-2-methoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, (5R)-5-[3-[(3S)-4-(4-fluoro-3-methoxy-phenyl) -3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, (5R)-5-[3-[4-(4,5-dichloro-2-methoxy-phenyl) piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, (5R)-5-[3-[(3S)-4-(4-chloro-2-methoxy-5-methyl-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, (5R)-5-[3-[(3S)-4-(3-chloro-5-methoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, (5R)-5-[3-[4-(4-chloro-2,5-dimethoxy-phenyl) piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, 2-[4-[3-[(3S) -4- (4-chloro-3-ethyl-phenyl) -3-methyl-piperazin-1-yl]-3-oxo-propyl]-2,5-dioxo-imidazolidin-4-yl]acetic acid, (5R)-5-[3-[(3S)-4-[4-chloro-3-(dimethylamino)-5-methoxy-phenyl]-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, 5-[3-[(3S) -4-(4-chloro-3-ethyl-phenyl) -3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-(6-methyl-2-pyridyl)imidazolidine-2,4-dione, 5-[3-[(3S)-4-[4-chloro-3-(trifluoromethoxy)phenyl]-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-(6-methyl-2-pyridyl) imidazolidine-2,4-dione, (5R) -5-[3-[4-(4-chloro-5-ethyl-2-methoxy-phenyl) piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, (5R)-5-[3-[(3S)-4-(4-chloro-5-ethyl-2-methoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, (5R)-5-[3-[(3S)-4-(4-chloro-2,5-dimethoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, (5R) -5-[3-[(3S) -4-(4-chloro-3-ethyl-5-methoxy-phenyl) -3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, (5R)-5-[3-[(3S)-4-(4-chloro-3-methoxy-phenyl)-3-ethyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, (5R)-5-[3-[(3S)-4-[5-methoxy-2-(trifluoromethoxy) phenyl]-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, (5R)-5-[3-[(3S)-4-(4-chloro-3-methoxy-phenyl)-3-isopropyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, (5R)-5-[3-[(3S)-4-(4-chloro-3,5-dimethoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, (5R) -5-[3-[(3S)-4-(4-chloro-3-ethoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, (5R)-5-[3-[4-(4-chloro-3-methoxy-5-methyl-phenyl) piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, (5R)-5-[3-[(3S)-4-(4,5-difluoro-2-methoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, tert-butyl 3-[4-[3-[(3S)-4-(4-chloro-3-ethyl-phenyl) -3-methyl-piperazin-1-yl]-3-oxo-propyl]-2,5-dioxo-imidazolidin-4-yl]propanoate, (5R) -5-[3-[4-(4-chloro-3-ethyl-5-methoxy-phenyl) piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, 3-[4-[3-[(3S)-4-(4-chloro-3-ethyl-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-2,5-dioxo-imidazolidin-4-yl]propanoic acid, 5-[3-[(3S)-4-(4-chloro-3-ethyl-5-methoxy-phenyl) -3-methyl-piperazin-1-yl]-2-methyl-3-oxo-propyl]-5-(methoxymethyl) imidazolidine-2,4-dione, (5R)-5-[3-[4-(4-chloro-3-methoxy-phenyl)-3-(trifluoromethyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, (5R)-5-[3-[(3S)-4-(4-chloro-3-fluoro-5-methoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, 5-[3-[(3S)-4-(4-chloro-3-fluoro-5-methoxy-phenyl) -3-methyl-piperazin-1-yl]-2-methyl-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, 5-[3-[(3S)-4-(4-chloro-3-fluoro-5-methoxy-phenyl) -3-methyl-piperazin-1-yl]-2-methyl-3-oxo-propyl]-5-(methoxymethyl) imidazolidine-2,4-dione, (5R)-5-[(2S)-3-[4-(4-chloro-3-fluoro-5-methoxy-phenyl)piperazin-1-yl]-2-methyl-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, (5S)-5-[(2S)-3-[4-(4-chloro-3-fluoro-5-methoxy-phenyl)piperazin-1-yl]-2-methyl-3-oxo-propyl]-5-(methoxymethyl) imidazolidine-2,4-dione, 5-[3-[(3S)-4-(4-chloro-3-ethyl-phenyl)-3-methyl-piperazin-1-yl]-2-methyl-3-oxo-propyl]-5-(methoxymethyl) imidazolidine-2,4-dione, 5-[3-[(3S)-4-(4-chloro-3-methoxy-5-methyl-phenyl)-3-methyl-piperazin-1-yl]-2-methyl-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, 5-[3-[(3S)-4-(4-chloro-3-methoxy-5-methyl-phenyl)-3-methyl-piperazin-1-yl]-2-methyl-3-oxo-propyl]-5-(methoxymethyl) imidazolidine-2,4-dione, (5R)-5-[3-[(3R)-4-(3,4-difluoro-5-methoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]imidazolidine-2,4-dione, (5R)-5-[3-[4-(3,4-difluoro-5-methoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]imidazolidine-2,4-dione, (5R)-5-[3-[(3S)-4-(3,4-difluoro-5-methoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]imidazolidine-2,4-dione, 5-cyclopropyl-5-[3-[4-(3-fluoro-5-methoxy-phenyl)piperazin-1-yl]-2-methyl-3-oxo-propyl]imidazolidine-2,4-dione, 5-[3-[4-(3-fluoro-5-methoxy-phenyl)piperazin-1-yl]-2-methyl-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, (5R)-5-[3-[(3S)-4- (3,5-dimethoxyphenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]imidazolidine-2,4-dione, (5R)-5-[3-[(3S)-4-(4-chloro-3-methoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]imidazolidine-2,4-dione, 5-cyclopropyl-5-[3-[(3S) -4-(3,4-difluoro-5-methoxy-phenyl)-3-methyl-piperazin-1-yl]-2-methyl-3-oxo-propyl]imidazolidine-2,4-dione, (5R)-5-[3-[4-(4-chloro-3,5-dimethoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]imidazolidine-2,4-dione, (5R)-5-[3-[4-(5-methoxy-2-methyl-phenyl)piperazin-1-yl]-3-oxo-propyl]imidazolidine-2,4-dione, (5R)-5-[3-[4-(3-methoxy-2-methyl-phenyl)piperazin-1-yl]-3-oxo-propyl]imidazolidine-2,4-dione, (5R)-5-[3-[4-(4-chloro-5-methoxy-2-methyl-phenyl)piperazin-1-yl]-3-oxo-propyl]imidazolidine-2,4-dione, (5R)-5-[3-[(3S)-4-(4-chloro-3,5-dimethoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]imidazolidine-2,4-dione, (5R)-5-[3-[(3S)-4-(5-methoxy-2-methyl-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]imidazolidine-2,4-dione, (5R)-5-[3-[(3S)-4-(3-methoxy-2-methyl-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]imidazolidine-2,4-dione, (5R)-5-[3-[(3S)-4-(4-chloro-5-methoxy-2-methyl-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]imidazolidine-2,4-dione, (5R)-5-[3-[4-(4-chloro-3-fluoro-5-methoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]imidazolidine-2,4-dione, (5R)-5-[3-[(3S)-4-(4-chloro-3-fluoro-5-methoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]imidazolidine-2,4-dione, (5R)-5-[3-[4-(3,4-dichloro-5-methoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]imidazolidine-2,4-dione, (5S)-5-[3-[(3S)-4-(4-chloro-3-methoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, (5R)-5-[3-[4-(3-chloro-4-fluoro-5-methoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]imidazolidine-2,4-dione, (5R)-5-[3-[(3S)-4-(3-chloro-4-fluoro-5-methoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]imidazolidine-2,4-dione, (5R)-5-[3-[4-(4-chloro-3-ethoxy-5-methoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]imidazolidine-2,4-dione, (5R)-5-[3-[(3S)-4-(4-chloro-3-ethoxy-5-methoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]imidazolidine-2,4-dione, or (5R)-5-[3-[4-(4-fluoro-3,5-dimethoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]imidazolidine-2,4-dione.
13. A pharmaceutical composition comprising a compound or pharmaceutically acceptable salt thereof according to claim 1, and a pharmaceutically acceptable carrier.
14. (canceled)
15. A method of treating and/or preventing inflammatory diseases, and/or diseases involving degradation of cartilage and/or disruption of cartilage homeostasis in a human comprising administering an effective amount of the compound or pharmaceutically acceptable salt thereof according to claim 1, to said human.
16. A method of treating and/or preventing inflammatory diseases, and/or diseases involving degradation of cartilage and/or disruption of cartilage homeostasis in a human comprising administering an effective amount of the pharmaceutical composition according to claim 13, to said human.
Description
DETAILED DESCRIPTION OF THE INVENTION
Definitions
[0079] The following terms are intended to have the meanings presented therewith below and are useful in understanding the description and intended scope of the present invention.
[0080] When describing the invention, which may include compounds, pharmaceutical compositions containing such compounds and methods of using such compounds and compositions, the following terms, if present, have the following meanings unless otherwise indicated. It should also be understood that when described herein any of the moieties defined forth below may be substituted with a variety of substituents, and that the respective definitions are intended to include such substituted moieties within their scope as set out below. Unless otherwise stated, the term substituted is to be defined as set out below. It should be further understood that the terms groups and radicals can be considered interchangeable when used herein.
[0081] The articles a and an may be used herein to refer to one or to more than one (i.e. at least one) of the grammatical objects of the article. By way of example an analogue means one analogue or more than one analogue.
[0082] Alkyl means straight or branched aliphatic hydrocarbon having the specified number of carbon atoms. Particular alkyl groups have 1 to 6 carbon atoms or 1 to 4 carbon atoms. Branched means that one or more alkyl groups such as methyl, ethyl or propyl is attached to a linear alkyl chain. Particular alkyl groups are methyl (CH.sub.3), ethyl (CH.sub.2CH.sub.3), n-propyl (CH.sub.2CH.sub.2CH.sub.3), isopropyl (CH(CH.sub.3).sub.2), n-butyl (CH.sub.2CH.sub.2CH.sub.2CH.sub.3), tert-butyl (CH.sub.2C(CH.sub.3).sub.3), sec-butyl (CH.sub.2CH(CH.sub.3).sub.2), n-pentyl (CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.3), n-hexyl (CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.3), and 1,2-dimethylbutyl (CHCH.sub.3)C(CH.sub.3)H.sub.2CH.sub.2CH.sub.3). Particular alkyl groups have between 1 and 4 carbon atoms.
[0083] Alkenyl refers to monovalent olefinically (unsaturated) hydrocarbon groups with the number of carbon atoms specified. Particular alkenyl has 2 to 8 carbon atoms, and more particularly, from 2 to 6 carbon atoms, which can be straight-chained or branched and having at least 1 and particularly from 1 to 2 sites of olefinic unsaturation. Particular alkenyl groups include ethenyl (CHCH.sub.2), n-propenyl (CH.sub.2CHCH.sub.2), isopropenyl (C(CH.sub.3)CH.sub.2) and the like.
[0084] Alkylene refers to divalent alkene radical groups having the number of carbon atoms specified, in particular having 1 to 6 carbon atoms and more particularly 1 to 4 carbon atoms which can be straight-chained or branched. This term is exemplified by groups such as methylene (CH.sub.2), ethylene (CH.sub.2CH.sub.2), or CH(CH.sub.3) and the like.
[0085] Alkynylene refers to divalent alkyne radical groups having the number of carbon atoms and the number of triple bonds specified, in particular 2 to 6 carbon atoms and more particularly 2 to 4 carbon atoms which can be straight-chained or branched. This term is exemplified by groups such as CC, CH.sub.2CC, and C(CH.sub.3)HCCH.
[0086] Alkoxy refers to the group O-alkyl, where the alkyl group has the number of carbon atoms specified. In particular the term refers to the group OC.sub.1-6 alkyl. Particular alkoxy groups are methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, tert-butoxy, sec-butoxy, n-pentoxy, n-hexoxy, and 1,2-dimethylbutoxy. Particular alkoxy groups are lower alkoxy, i.e. with between 1 and 6 carbon atoms. Further particular alkoxy groups have between 1 and 4 carbon atoms.
[0087] Amino refers to the radical NH.sub.2.
[0088] Aryl refers to a monovalent aromatic hydrocarbon group derived by the removal of one hydrogen atom from a single carbon atom of a parent aromatic ring system. In particular aryl refers to an aromatic ring structure, monocyclic or fused polycyclic, with the number of ring atoms specified. Specifically, the term includes groups that include from 6 to 10 ring members. Particular aryl groups include phenyl, and naphthyl.
[0089] Cycloalkyl refers to a non-aromatic hydrocarbyl ring structure, monocyclic, fused polycyclic, bridged polycyclic, or spirocyclic, with the number of ring atoms specified. A cycloalkyl may have from 3 to 12 carbon atoms, in particular from 3 to 10, and more particularly from 3 to 7 carbon atoms. Such cycloalkyl groups include, by way of example, single ring structures such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl.
[0090] Cyano refers to the radical CN.
[0091] Halo or halogen refers to fluoro (F), chloro (Cl), bromo (Br) and iodo (I). Particular halo groups are either fluoro or chloro.
[0092] Hetero when used to describe a compound or a group present on a compound means that one or more carbon atoms in the compound or group have been replaced by a nitrogen, oxygen, or sulfur heteroatom. Hetero may be applied to any of the hydrocarbyl groups described above such as alkyl, e.g. heteroalkyl, cycloalkyl, e.g. heterocycloalkyl, aryl, e.g. heteroaryl, and the like having from 1 to 4, and particularly from 1 to 3 heteroatoms, more typically 1 or 2 heteroatoms, for example a single heteroatom.
[0093] Heteroaryl means an aromatic ring structure, monocyclic or fused polycyclic, that includes one or more heteroatoms independently selected from O, N and S and the number of ring atoms specified. In particular, the aromatic ring structure may have from 5 to 9 ring members. The heteroaryl group can be, for example, a five membered or six membered monocyclic ring or a fused bicyclic structure formed from fused five and six membered rings or two fused six membered rings or, by way of a further example, two fused five membered rings. Each ring may contain up to four heteroatoms typically selected from nitrogen, sulphur and oxygen. Typically the heteroaryl ring will contain up to 4 heteroatoms, more typically up to 3 heteroatoms, more usually up to 2, for example a single heteroatom. In one embodiment, the heteroaryl ring contains at least one ring nitrogen atom. The nitrogen atoms in the heteroaryl rings can be basic, as in the case of an imidazole or pyridine, or essentially non-basic as in the case of an indole or pyrrole nitrogen. In general the number of basic nitrogen atoms present in the heteroaryl group, including any amino group substituents of the ring, will be less than five.
[0094] Examples of five membered monocyclic heteroaryl groups include but are not limited to pyrrolyl, furanyl, thiophenyl, imidazolyl, furazanyl, oxazolyl, oxadiazolyl, oxatriazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyrazolyl, triazolyl and tetrazolyl groups.
[0095] Examples of six membered monocyclic heteroaryl groups include but are not limited to pyridinyl, pyrazinyl, pyridazinyl, pyrimidinyl and triazinyl.
[0096] Particular examples of bicyclic heteroaryl groups containing a five membered ring fused to another five-membered ring include but are not limited to imidazothiazolyl and imidazoimidazolyl.
[0097] Particular examples of bicyclic heteroaryl groups containing a six membered ring fused to a five membered ring include but are not limited to benzofuranyl, benzothiophenyl, benzoimidazolyl, benzoxazolyl, isobenzoxazolyl, benzisoxazolyl, benzothiazolyl, benzoisothiazolyl, isobenzofuranyl, indolyl, isoindolyl, indolizinyl, purinyl (e.g. adenine, guanine), indazolyl, pyrazolopyrimidinyl, triazolopyrimidinyl, and pyrazolopyridinyl groups.
[0098] Particular examples of bicyclic heteroaryl groups containing two fused six membered rings include but are not limited to quinolinyl, isoquinolinyl, pyridopyridinyl, quinoxalinyl, quinazolinyl, cinnolinyl, phthalazinyl, naphthyridinyl, and pteridinyl groups. Particular heteroaryl groups are those derived from thiophenyl, pyrrolyl, benzothiophenyl, benzofuranyl, indolyl, pyridinyl, quinolinyl, imidazolyl, oxazolyl and pyrazinyl.
[0099] Examples of representative heteroaryls include the following:
##STR00003##
wherein each Y is selected from >CO, NH, O and S.
[0100] Heterocycloalkyl means a non-aromatic fully saturated ring structure, monocyclic, fused polycyclic, spirocyclic, or bridged polycyclic, that includes one or more heteroatoms independently selected from O, N and S and the number of ring atoms specified. The heterocycloalkyl ring structure may have from 4 to 12 ring members, in particular from 4 to 10 ring members and more particularly from 4 to 7 ring members. Each ring may contain up to four heteroatoms typically selected from nitrogen, sulphur and oxygen. Typically the heterocycloalkyl ring will contain up to 4 heteroatoms, more typically up to 3 heteroatoms, more usually up to 2, for example a single heteroatom. Examples of heterocyclic rings include, but are not limited to azetidinyl, oxetanyl, thietanyl, pyrrolidinyl (e.g. 1-pyrrolidinyl, 2-pyrrolidinyl and 3-pyrrolidinyl), tetrahydrofuranyl (e.g. 1-tetrahydrofuranyl, 2-tetrahydrofuranyl and 3-tetrahydrofuranyl), tetrahydrothiophenyl (e.g. 1-tetrahydrothiophenyl, 2-tetrahydrothiophenyl and 3-tetrahydrothiophenyl), piperidinyl (e.g. 1-piperidinyl, 2-piperidinyl, 3-piperidinyl and 4-piperidinyl), tetrahydropyranyl (e.g. 4-tetrahydropyranyl), tetrahydrothiopyranyl (e.g. 4-tetrahydrothiopyranyl), morpholinyl, thiomorpholinyl, dioxanyl, or piperazinyl.
[0101] As used herein, the term heterocycloalkenyl means a heterocycloalkyl, which comprises at least one double bond. Particular examples of heterocycloalkenyl groups are shown in the following illustrative examples:
##STR00004##
wherein each W is selected from CH.sub.2, NH, O and S; each Y is selected from NH, O, C(O), SO.sub.2, and S; and each Z is selected from N or CH.
[0102] Particular examples of monocyclic rings are shown in the following illustrative examples:
##STR00005##
wherein each W and Y is independently selected from CH.sub.2, NH, O and S.
[0103] Particular examples of fused bicyclic rings are shown in the following illustrative examples:
##STR00006##
wherein each W and Y is independently selected from CH.sub.2, NH, O and S.
[0104] Particular examples of bridged bicyclic rings are shown in the following illustrative examples:
##STR00007##
wherein each W and Y is independently selected from CH.sub.2, NH, O and S and each Z is selected from N and CH.
[0105] Particular examples of spirocyclic rings are shown in the following illustrative examples:
##STR00008##
wherein each Y is selected from CH.sub.2, NH, O and S.
[0106] Hydroxyl refers to the radical OH.
[0107] Oxo refers to the radical O.
[0108] Substituted refers to a group in which one or more hydrogen atoms are each independently replaced with the same or different substituent(s).
[0109] Sulfo or sulfonic acid refers to a radical such as SO.sub.3H.
[0110] Thiol refers to the group SH.
[0111] As used herein, term substituted with one or more refers to one to four substituents. In one embodiment it refers to one to three substituents. In further embodiments it refers to one or two substituents. In a yet further embodiment it refers to one substituent.
[0112] Thioalkoxy refers to the group S-alkyl where the alkyl group has the number of carbon atoms specified. In particular the term refers to the group SC.sub.1-6 alkyl. Particular thioalkoxy groups are thiomethoxy, thioethoxy, n-thiopropoxy, isothiopropoxy, n-thiobutoxy, tert-thiobutoxy, sec-thiobutoxy, n-thiopentoxy, n-thiohexoxy, and 1,2-dimethylthiobutoxy. Particular thioalkoxy groups are lower thioalkoxy, i.e. with between 1 and 6 carbon atoms. Further particular alkoxy groups have between 1 and 4 carbon atoms.
[0113] One having ordinary skill in the art of organic synthesis will recognize that the maximum number of heteroatoms in a stable, chemically feasible heterocyclic ring, whether it is aromatic or non-aromatic, is determined by the size of the ring, the degree of unsaturation and the valence of the heteroatoms. In general, a heterocyclic ring may have one to four heteroatoms so long as the heteroaromatic ring is chemically feasible and stable.
[0114] Pharmaceutically acceptable means approved or approvable by a regulatory agency of the Federal or a state government or the corresponding agency in countries other than the United States, or that is listed in the U.S. Pharmacopoeia or other generally recognized pharmacopoeia for use in animals, and more particularly, in humans.
[0115] Pharmaceutically acceptable salt refers to a salt of a compound of the invention that is pharmaceutically acceptable and that possesses the desired pharmacological activity of the parent compound. In particular, such salts are non-toxic may be inorganic or organic acid addition salts and base addition salts. Specifically, such salts include: (1) acid addition salts, formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like; or formed with organic acids such as acetic acid, propionic acid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, 3-(4-hydroxybenzoyl) benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethane-disulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, 4-toluenesulfonic acid, camphorsulfonic acid, 4-methylbicyclo[2.2.2]-oct-2-ene-1-carboxylic acid, glucoheptonic acid, 3-phenylpropionic acid, trimethylacetic acid, tertiary butylacetic acid, lauryl sulfuric acid, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid, muconic acid, and the like; or (2) salts formed when an acidic proton present in the parent compound either is replaced by a metal ion, e.g. an alkali metal ion, an alkaline earth ion, or an aluminum ion; or coordinates with an organic base such as ethanolamine, diethanolamine, triethanolamine, N-methylglucamine and the like. Salts further include, by way of example only, sodium, potassium, calcium, magnesium, ammonium, tetraalkylammonium, and the like; and when the compound contains a basic functionality, salts of non-toxic organic or inorganic acids, such as hydrochloride, hydrobromide, tartrate, mesylate, acetate, maleate, oxalate and the like. The term pharmaceutically acceptable cation refers to an acceptable cationic counter-ion of an acidic functional group. Such cations are exemplified by sodium, potassium, calcium, magnesium, ammonium, tetraalkylammonium cations, and the like.
[0116] Pharmaceutically acceptable vehicle refers to a diluent, adjuvant, excipient or carrier with which a compound of the invention is administered.
[0117] Prodrugs refers to compounds, including derivatives of the compounds of the invention, which have cleavable groups and become by solvolysis or under physiological conditions the compounds of the invention which are pharmaceutically active in vivo. Such examples include, but are not limited to, choline ester derivatives and the like, N-alkylmorpholine esters and the like.
[0118] Solvate refers to forms of the compound that are associated with a solvent, usually by a solvolysis reaction. This physical association includes hydrogen bonding. Conventional solvents include water, EtOH, acetic acid and the like. The compounds of the invention may be prepared e.g. in crystalline form and may be solvated or hydrated. Suitable solvates include pharmaceutically acceptable solvates, such as hydrates, and further include both stoichiometric solvates and non-stoichiometric solvates. In certain instances the solvate will be capable of isolation, for example when one or more solvent molecules are incorporated in the crystal lattice of the crystalline solid. Solvate encompasses both solution-phase and isolable solvates. Representative solvates include hydrates, ethanolates and methanolates.
[0119] Subject includes humans. The terms human, patient and subject are used interchangeably herein.
[0120] Effective amount means the amount of a compound of the invention that, when administered to a subject for treating a disease, is sufficient to effect such treatment for the disease. The effective amount can vary depending on the compound, the disease and its severity, and the age, weight, etc., of the subject to be treated.
[0121] Preventing or prevention refers to a reduction in risk of acquiring or developing a disease or disorder (i.e. causing at least one of the clinical symptoms of the disease not to develop in a subject that may be exposed to a disease-causing agent, or predisposed to the disease in advance of disease onset.
[0122] The term prophylaxis is related to prevention, and refers to a measure or procedure the purpose of which is to prevent, rather than to treat or cure a disease. Non-limiting examples of prophylactic measures may include the administration of vaccines; the administration of low molecular weight heparin to hospital patients at risk for thrombosis due, for example, to immobilization; and the administration of an anti-malarial agent such as chloroquine, in advance of a visit to a geographical region where malaria is endemic or the risk of contracting malaria is high.
[0123] Treating or treatment of any disease or disorder refers, in one embodiment, to ameliorating the disease or disorder (i.e. arresting the disease or reducing the manifestation, extent or severity of at least one of the clinical symptoms thereof). In another embodiment treating or treatment refers to ameliorating at least one physical parameter, which may not be discernible by the subject. In yet another embodiment, treating or treatment refers to modulating the disease or disorder, either physically, (e.g. stabilization of a discernible symptom), physiologically, (e.g. stabilization of a physical parameter), or both. In a further embodiment, treating or treatment relates to slowing the progression of the disease.
[0124] As used herein the term inflammatory diseases refers to the group of conditions including rheumatoid arthritis, osteoarthritis, juvenile idiopathic arthritis, psoriasis, psoriatic arthritis, allergic airway disease (e.g. asthma, rhinitis), chronic obstructive pulmonary disease (COPD), inflammatory bowel diseases (e.g. Crohn's disease, ulcerative colitis), endotoxin-driven disease states (e.g. complications after bypass surgery or chronic endotoxin states contributing to e.g. chronic cardiac failure), and related diseases involving cartilage, such as that of the joints. Particularly the term refers to rheumatoid arthritis, osteoarthritis, allergic airway disease (e.g. asthma), chronic obstructive pulmonary disease (COPD) and inflammatory bowel diseases. More particularly the term refers to rheumatoid arthritis, and osteoarthritis (OA). Most particularly the term refers to osteoarthritis (OA).
[0125] As used herein the term allergic disease(s) refers to the group of conditions characterized by a hypersensitivity disorder of the immune system including, allergic airway disease (e.g. asthma, rhinitis), sinusitis, eczema and hives, as well as food allergies or allergies to insect venom.
[0126] As used herein the term asthma as used herein refers to any disorder of the lungs characterized by variations in pulmonary gas flow associated with airway constriction of whatever cause (intrinsic, extrinsic, or both; allergic or non-allergic). The term asthma may be used with one or more adjectives to indicate the cause.
[0127] As used herein the term diseases involving degradation of cartilage and/or disruption of cartilage homeostasis includes conditions such as osteoarthritis, psoriatic arthritis, juvenile rheumatoid arthritis, gouty arthritis, septic or infectious arthritis, reactive arthritis, reflex sympathetic dystrophy, algodystrophy, achondroplasia, Paget's disease, Tietze syndrome or costal chondritis, fibromyalgia, osteochondritis, neurogenic or neuropathic arthritis, arthropathy, sarcoidosis, amylosis, hydarthrosis, periodical disease, rheumatoid spondylitis, endemic forms of arthritis like osteoarthritis deformans endemica, Mseleni disease and Handigodu disease; degeneration resulting from fibromyalgia, systemic lupus erythematosus, scleroderma and ankylosing spondylitis. More particularly, the term refers to osteoarthritis (OA).
[0128] Compound(s) of the invention, and equivalent expressions, are meant to embrace compounds of the Formula(e) as herein described, which expression includes the pharmaceutically acceptable salts, and the solvates, e.g. hydrates, and the solvates of the pharmaceutically acceptable salts where the context so permits. Similarly, reference to intermediates, whether or not they themselves are claimed, is meant to embrace their salts, and solvates, where the context so permits.
[0129] When ranges are referred to herein, for example but without limitation, C.sub.1-8 alkyl, the citation of a range should be considered a representation of each member of said range.
[0130] Other derivatives of the compounds of this invention have activity in both their acid and acid derivative forms, but in the acid sensitive form often offers advantages of solubility, tissue compatibility, or delayed release in the mammalian organism [1]. Prodrugs include acid derivatives well known to practitioners of the art, such as, for example, esters prepared by reaction of the parent acid with a suitable alcohol, or amides prepared by reaction of the parent acid compound with a substituted or unsubstituted amine, or acid anhydrides, or mixed anhydrides. Simple aliphatic or aromatic esters, amides and anhydrides derived from acidic groups pendant on the compounds of this invention are particularly useful prodrugs. In some cases it is desirable to prepare double ester type prodrugs such as (acyloxy)alkyl esters or ((alkoxycarbonyl)oxy)alkylesters. Particular such prodrugs are the C.sub.1-8 alkyl, C.sub.2-8 alkenyl, C.sub.6-1.sub.0 optionally substituted aryl, and (C.sub.6-10 aryl)-(C.sub.1-4 alkyl) esters of the compounds of the invention.
[0131] As used herein, the term isotopic variant refers to a compound that contains unnatural proportions of isotopes at one or more of the atoms that constitute such compound. For example, an isotopic variant of a compound can contain one or more non-radioactive isotopes, such as for example, deuterium (.sup.2H or D), carbon-13 (.sup.13C), nitro (.sup.15N), or the like. It will be understood that, in a compound where such isotopic substitution is made, the following atoms, where present, may vary, so that for example, any hydrogen may be .sup.2H/D, any carbon may be .sup.13C, or any nitrogen may be .sup.15N, and that the presence and placement of such atoms may be determined within the skill of the art. Likewise, the invention may include the preparation of isotopic variants with radioisotopes, in the instance for example, where the resulting compounds may be used for drug and/or substrate tissue distribution studies. The radioactive isotopes tritium, i.e. .sup.3H, and carbon-14, i.e. .sup.14C, are particularly useful for this purpose in view of their ease of incorporation and ready means of detection. Further, compounds may be prepared that are substituted with positron emitting isotopes, such as .sup.11C, .sup.18F, .sup.15O and .sup.13N, and would be useful in Positron Emission Topography (PET) studies for examining substrate receptor occupancy.
[0132] It is also to be understood that compounds that have the same molecular formula but differ in the nature or sequence of bonding of their atoms or the arrangement of their atoms in space are termed isomers. Isomers that differ in the arrangement of their atoms in space are termed stereoisomers.
[0133] Stereoisomers that are not mirror images of one another are termed diastereomers and those that are non-superimposable mirror images of each other are termed enantiomers. When a compound has an asymmetric center, for example, it is bonded to four different groups, a pair of enantiomers is possible.
[0134] An enantiomer can be characterized by the absolute configuration of its asymmetric center and is described by the R- and S-sequencing rules of Cahn and Prelog, or by the manner in which the molecule rotates the plane of polarized light and designated as dextrorotatory or levorotatory (i.e. as (+) or ()-isomers respectively). A chiral compound can exist as either individual enantiomer or as a mixture thereof. A mixture containing equal proportions of the enantiomers is called a racemic mixture.
[0135] Tautomers refer to compounds that are interchangeable forms of a particular compound structure, and that vary in the displacement of hydrogen atoms and electrons. Thus, two structures may be in equilibrium through the movement of 71 electrons and an atom (usually H). For example, enols and ketones are tautomers because they are rapidly interconverted by treatment with either acid or base. Another example of tautomerism is the aci- and nitro-forms of phenylnitromethane, that are likewise formed by treatment with acid or base.
[0136] Tautomeric forms may be relevant to the attainment of the optimal chemical reactivity and biological activity of a compound of interest.
[0137] The compounds of the invention may possess one or more asymmetric centers; such compounds can therefore be produced as individual (R)- or (S)-stereoisomers or as mixtures thereof.
[0138] Unless indicated otherwise, the description or naming of a particular compound in the specification and claims is intended to include both individual enantiomers and mixtures, racemic or otherwise, thereof. The methods for the determination of stereochemistry and the separation of stereoisomers are well-known in the art.
[0139] It will be appreciated that compounds of the invention may be metabolized to yield biologically active metabolites.
The Invention
[0140] The present invention is based on the identification of novel hydantoin compounds that may be useful for the prophylaxis and/or treatment of inflammatory diseases, and/or diseases involving degradation of cartilage and/or disruption of cartilage homeostasis. In a particular aspect, the present compounds are ADAMTS inhibitors, particularly ADAMTS-5 and/or ADAMTS-6.
[0141] The present invention also provides methods for the production of these compounds, pharmaceutical compositions comprising these compounds and methods for treating inflammatory diseases, and/or diseases involving degradation of cartilage and/or disruption of cartilage homeostasis by administering the compounds of the invention.
[0142] Accordingly, in a first aspect of the invention, the compounds of the invention are provided having a Formula (I):
##STR00009##
R.SUP.1 .is:
[0143] H, [0144] C.sub.1-4 alkyl optionally substituted with one or more independently selected R.sup.4 groups, [0145] C.sub.3-7 monocyclic cycloalkyl optionally substituted with one or more independently selected R.sup.4 groups, [0146] 4-7 membered monocyclic heterocycloalkyl comprising 1 to 2 heteroatoms independently selected from N, O, and S, optionally substituted with one or more independently selected C.sub.1-4 alkyl, C(O)C.sub.1-4 alkyl, or C(O)OC.sub.1-4 alkyl, [0147] phenyl optionally substituted with one or more independently selected R.sup.5 groups, [0148] phenyl fused to a 5-6 membered monocyclic heterocycloalkyl comprising 1, 2 or 3 heteroatoms independently selected from N, O, and S, which heterocycloalkyl is optionally substituted with one or more O, [0149] 5-6 membered monocyclic heteroaryl comprising 1 or 2 heteroatoms independently selected from N, O, and S, optionally substituted with one or more independently selected R.sup.5 groups;
R.sup.2 is independently selected from: [0150] H, [0151] OH, [0152] C.sub.1-4 alkoxy, and [0153] C.sub.1-4 alkyl optionally substituted with one [0154] OH, [0155] CN, [0156] C.sub.1-4 alkoxy optionally substituted with one phenyl, and [0157] 5-6 membered monocyclic heteroaryl comprising 1 or 2 heteroatoms independently selected from N, O, and S, optionally substituted with one or more independently selected C.sub.1-4 alkyl;
each R.sup.3a, and R.sup.3b is independently selected from H, and C.sub.1-4 alkyl optionally substituted with one or more halo;
Cy is
[0158] 6-10 membered monocyclic or fused bicyclic aryl, [0159] 5-10 membered monocyclic or fused bicyclic heteroaryl comprising 1, 2 or 3 heteroatoms independently selected from N, O, and S;
R.SUP.4 .is
[0160] halo, [0161] OH, [0162] CN, [0163] C.sub.1-4 alkyl, [0164] C.sub.1-4 alkoxy optionally substituted with one C.sub.1-4 alkoxy, or phenyl, [0165] C.sub.1-4 thioalkoxy, [0166] 4-7-membered monocyclic heterocycloalkyl comprising one or more heteroatoms independently selected from N, S, and O, optionally substituted with one or more halo, or C(O)OC.sub.1-4 alkyl, [0167] phenyl, [0168] S(O).sub.2C.sub.1-4 alkyl [0169] C(O)OR.sup.7a [0170] C(O)NR.sup.7bR.sup.7c [0171] NHC(O)OR.sup.7d [0172] NHC(O)R.sup.7e [0173] NR.sup.8aR.sup.8b;
each R.sup.5 is [0174] halo, [0175] OH, [0176] CN, [0177] C.sub.1-4 alkyl optionally substituted with one or more independently selected halo, NR.sup.9aR.sup.9b, or C(O)NR.sup.9cR.sup.9d, [0178] C.sub.1-4 alkoxy optionally substituted with NR.sup.9eR.sup.9f, or S(O).sub.2C.sub.1-4 alkyl;
each R.sup.6a is [0179] C.sub.2-4 alkyl optionally substituted with one or more halo, [0180] C.sub.1-4 alkoxy optionally substituted with one or more halo,
the subscript n is 0, 1, 2 or 3
each R.sup.6b is independently selected from: [0181] halo, [0182] CN, [0183] NO.sub.2, [0184] C.sub.1-4 alkyl, [0185] C.sub.1-4 alkoxy [0186] 5-10 membered monocyclic or fused bicyclic heteroaryl comprising 1, 2 or 3 heteroatoms independently selected from N, O, and S, optionally substituted with one or more independently selected halo, C.sub.1-4 alkyl, or C.sub.1-4 alkoxy, and [0187] NR.sup.9gR.sup.9h;
each R.sup.7a, R.sup.7b, R.sup.7c, R.sup.7d, or R.sup.7e, is H, or C.sub.1-4 alkyl optionally substituted with OH or C.sub.1-4 alkoxy;
each R.sup.8a, or R.sup.8b is independently selected from H, and C.sub.1-4 alkyl optionally substituted with OH, C.sub.1-4 alkoxy, or phenyl;
each R.sup.9a, R.sup.9b, R.sup.9c, R.sup.9d, R.sup.9e, R.sup.9f, R.sup.9g, R.sup.9h is independently selected from H, and C.sub.1-4 alkyl;
or a pharmaceutically acceptable salt, or a solvate, or a pharmaceutically acceptable salt of a solvate thereof; or a biologically active metabolite thereof;
provided that when R.sup.1, R.sup.2, R.sup.3a and R.sup.3b are H, Cy is phenyl and R.sup.6a is OCH.sub.3, then the subscript n is not 0.
[0188] In one embodiment, a compound of the invention is according to Formula II:
##STR00010##
wherein R.sup.1, R.sup.2, R.sup.3a, R.sup.3b, R.sup.6a, R.sup.6b, the subscript n and Cy are as defined above.
[0189] In one embodiment, a compound of the invention is according to Formula I or II, wherein R.sup.1 is H.
[0190] In another embodiment, a compound of the invention is according to Formula I or II, wherein R.sup.1 is C.sub.1-4 alkyl. In a particular embodiment, R.sup.1 is Me, Et, Pr, iPr, or tBu. In a more particular embodiment, R.sup.1 is Me, or Et. In a particular embodiment, R.sup.1 is Me.
[0191] In another embodiment, a compound of the invention is according to Formula I or II, wherein R.sup.1 is C.sub.1-4 alkyl substituted with one or more independently selected R.sup.4 groups. In another embodiment, R.sup.1 is Me, or Et, each of which is substituted with one or more independently selected R.sup.4 groups. In a particular embodiment, R.sup.1 is C.sub.1-4 alkyl substituted with one, two or three independently selected R.sup.4 groups. In another particular embodiment, R.sup.1 is Me, or Et, each of which is substituted with one, two or three independently selected R.sup.4 groups. In a more particular embodiment, R.sup.1 is C.sub.1-4 alkyl substituted with one R.sup.4 group. In another more particular embodiment, R.sup.1 is Me, or Et, each of which is substituted with one R.sup.4 group. In a most particular embodiment, R.sup.1 is CH.sub.2R.sub.4.
[0192] In one embodiment, a compound of the invention is according to Formula I or II, wherein R.sup.1 is C.sub.3-7 monocyclic cycloalkyl. In a particular embodiment, R.sup.1 is cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl. In a more particular embodiment, R.sup.1 is cyclopropyl.
[0193] In another embodiment, a compound of the invention is according to Formula I or II, wherein R.sup.1 is C.sub.3-7 monocyclic cycloalkyl substituted with one or more independently selected R.sup.4 groups. In another embodiment, R.sup.1 is cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl, each of which is substituted with one or more independently selected R.sup.4 groups. In a particular embodiment, R.sup.1 is C.sub.3-7 monocyclic cycloalkyl substituted with one, two or three independently selected R.sup.4 groups. In another particular embodiment, R.sup.1 is cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl, each of which is substituted with one, two or three independently selected R.sup.4 groups. In a more particular embodiment, R.sup.1 is C.sub.3-7 monocyclic cycloalkyl substituted with one R.sup.4 group. In another more particular embodiment, R.sup.1 is cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl, each of which is substituted with one R.sup.4 group.
[0194] In one embodiment, a compound of the invention is according to Formula I or II, wherein R.sup.4 is halo, OH, and CN. In a more particular embodiment, each R.sup.4 is independently selected from F, Cl, OH, and CN.
[0195] In one embodiment, a compound of the invention is according to Formula I or II, wherein R.sup.4 is C.sub.1-4 alkyl. In a particular embodiment, R.sup.4 is CH.sub.3, CH.sub.2CH.sub.3, or CH(CH.sub.3).sub.2. In a more particular embodiment, R.sup.4 is CH.sub.3.
[0196] In one embodiment, a compound of the invention is according to Formula I or II, wherein R.sup.4 is C.sub.1-4 alkoxy. In a particular embodiment, R.sup.4 is OMe, OEt, or OiPr. In a more particular embodiment, R.sup.4 is OMe.
[0197] In one embodiment, a compound of the invention is according to Formula I or II, wherein R.sup.4 is C.sub.1-4 alkoxy substituted with one C.sub.1-4 alkoxy, or phenyl. In a particular embodiment, R.sup.4 is OMe, OEt, or OiPr, each of which is substituted with one C.sub.1-4 alkoxy, or phenyl. In a more particular embodiment, R.sup.4 is C.sub.1-4 alkoxy substituted with one OMe, OEt, or phenyl. In another more particular embodiment, R.sup.4 is OMe, OEt, or OiPr, each of which is substituted with one OMe, OEt, or phenyl. In a most particular embodiment, R.sup.4 is OCH.sub.2CH.sub.2OCH.sub.3, OCH.sub.2Ph.
[0198] In one embodiment, a compound of the invention is according to Formula I or II, wherein R.sup.4 is C.sub.1-4 thioalkoxy. In a particular embodiment, R.sup.4 is SCH.sub.3, or SCH.sub.2CH.sub.3. In a more particular embodiment, R.sup.4 is SCH.sub.3.
[0199] In one embodiment, a compound of the invention is according to Formula I or II, wherein R.sup.4 is 4-7-membered monocyclic heterocycloalkyl comprising one or more heteroatoms independently selected from N, S, and O. In a particular embodiment, R.sup.4 is azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, tetrahydrofuranyl, tetrahydropyranyl, or dioxanyl. In a more particular embodiment, R.sup.4 is azetidinyl, pyrrolidinyl, piperidinyl, or morpholinyl.
[0200] In one embodiment, a compound of the invention is according to Formula I or II, wherein R.sup.4 is 4-7-membered monocyclic heterocycloalkyl comprising one or more heteroatoms independently selected from N, S, and O, substituted with one or more halo, C(O)OC.sub.1_.sub.4 alkyl. In a particular embodiment, R.sup.4 is 4-7-membered monocyclic heterocycloalkyl comprising one or more heteroatoms independently selected from N, S, and O, substituted with one, two or three independently selected F, Cl, C(O)OCH.sub.3, C(O)OCH.sub.2CH.sub.3, or C(O)OC(CH.sub.3).sub.3. In another particular embodiment, R.sup.4 is azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, tetrahydrofuranyl, tetrahydropyranyl, or dioxanyl, each of which is substituted with one, two or three independently selected F, Cl, C(O)OCH.sub.3, C(O)OCH.sub.2CH.sub.3, or C(O)OC(CH.sub.3).sub.3. In a more particular embodiment, R.sup.4 is 4-7-membered monocyclic heterocycloalkyl comprising one or more heteroatoms independently selected from N, S, and O, substituted with one F, Cl, C(O)OCH.sub.3, C(O)OCH.sub.2CH.sub.3, or C(O)OC(CH.sub.3).sub.3. In another particular embodiment, R.sup.4 is azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, tetrahydrofuranyl, tetrahydropyranyl, or dioxanyl, each of which is substituted with one F, Cl, C(O)OCH.sub.3, C(O)OCH.sub.2CH.sub.3, or C(O)OC(CH.sub.3).sub.3. In a most particular embodiment, R.sup.4 is azetidinyl, pyrrolidinyl, piperidinyl, or morpholinyl, each of which is substituted with one, two or three independently selected F, Cl. In another most particular embodiment, R.sup.4 is azetidinyl, pyrrolidinyl, piperidinyl, or morpholinyl, each of which is substituted with one C(O)OCH.sub.3, C(O)OCH.sub.2CH.sub.3, or C(O)OC(CH.sub.3).sub.3.
[0201] In one embodiment, a compound of the invention is according to Formula I or II, wherein R.sup.4 is phenyl.
[0202] In one embodiment, a compound of the invention is according to Formula I or II, wherein R.sup.4 is S(O).sub.2C.sub.1-4 alkyl. In a particular embodiment, R.sup.4 is S(O).sub.2CH.sub.3, or S(O).sub.2CH.sub.2CH.sub.3.
[0203] In one embodiment, a compound of the invention is according to Formula I or II, wherein R.sup.4 is C(O)OR.sup.7a, and R.sup.7a is as previously described. In a particular embodiment, R.sup.7a is H. In another particular embodiment, R.sup.7a is C.sub.1-4 alkyl. In yet another particular embodiment, R.sup.7a is C.sub.1-4 alkyl substituted with one OH, C.sub.1-4 alkoxy. In a more particular embodiment, R.sup.7a is Me, Et, iPr or tBu. In another more particular embodiment, R.sup.7a is Me, Et, iPr or tBu, each of which is substituted with one OH, C.sub.1-4 alkoxy. In yet another more particular embodiment, R.sup.7a is Me, Et, iPr or tBu, each of which is substituted with one OH, OCH.sub.3. In a most particular embodiment, R.sup.4 is C(O)OCH.sub.3, C(O)OCH.sub.2CH.sub.3, or C(O)OC(CH.sub.3).sub.3.
[0204] In one embodiment, a compound of the invention is according to Formula I or II, wherein R.sup.4 is C(O)NR.sup.7bR.sup.7c, and each R.sup.7b or R.sup.7c is as previously described. In a particular embodiment, R.sup.7b and R.sup.7c are H. In another particular embodiment, one of R.sup.7b or R.sup.7e is H, and the other is C.sub.1-4 alkyl. In yet another particular embodiment, one of R.sup.7b or R.sup.7c is H, and the other is C.sub.1-4 alkyl substituted with one OH, C.sub.1-4 alkoxy. In a further particular embodiment, R.sup.7b and R.sup.7c are C.sub.1-4 alkyl. In a more particular embodiment, one of R.sup.7b or R.sup.7c is H, and the other is Me, Et, iPr or tBu. In another more particular embodiment, one of R.sup.7b or R.sup.7c is H, and the other is Me, Et, iPr or tBu, each of which is substituted with one OH, C.sub.1-4 alkoxy. In yet another more particular embodiment, one of R.sup.7b or R.sup.7c is H, and the other is Me, Et, iPr or tBu, each of which is substituted with one OH, OCH.sub.3. In a most particular embodiment, R.sup.4 is C(O)NHCH.sub.3, C(O)N(CH.sub.3).sub.2, C(O)NHCH.sub.2CH.sub.3, C(O)NHCH.sub.2CH.sub.2OH or C(O)NHCH.sub.2CH.sub.2OCH.sub.3.
[0205] In one embodiment, a compound of the invention is according to Formula I or II, wherein R.sup.4 is NHC(O)OR.sup.7d, and R.sup.7d is as previously described. In a particular embodiment, R.sup.7d is H. In another particular embodiment, R.sup.7d is C.sub.1-4 alkyl. In yet another particular embodiment, R.sup.7d is C.sub.1-4 alkyl substituted with one OH, C.sub.1-4 alkoxy. In a more particular embodiment, R.sup.7d is Me, Et, iPr or tBu. In another more particular embodiment, R.sup.7d is Me, Et, iPr or tBu, each of which is substituted with one OH, C.sub.1-4 alkoxy. In yet another more particular embodiment, R.sup.7d is Me, Et, iPr or tBu, each of which is substituted with one OH, OCH.sub.3. In a most particular embodiment, R.sup.4 is NHC(O)OCH.sub.3, NHC(O)OCH.sub.2CH.sub.3, or NHC(O)OC(CH.sub.3).sub.3.
[0206] In one embodiment, a compound of the invention is according to Formula I or II, wherein R.sup.4 is NHC(O)R.sup.7e, and R.sup.7e is as previously described. In a particular embodiment, R.sup.7e is H. In another particular embodiment, R.sup.7e is C.sub.1-4 alkyl. In yet another particular embodiment, R.sup.7e is C.sub.1-4 alkyl substituted with one OH, C.sub.1-4 alkoxy. In a more particular embodiment, R.sup.7e is Me, Et, iPr or tBu. In another more particular embodiment, R.sup.7e is Me, Et, iPr or tBu, each of which is substituted with one OH, C.sub.1-4 alkoxy. In yet another more particular embodiment, R.sup.7e is Me, Et, iPr or tBu, each of which is substituted with one OH, OCH.sub.3. In a most particular embodiment, R.sup.4 is NHC(O)CH.sub.3, NHC(O)CH.sub.2CH.sub.3, or NHC(O)C(CH.sub.3).sub.3.
[0207] In one embodiment, a compound of the invention is according to Formula I or II, wherein R.sup.4 is NR.sup.8aR.sup.8b, and each R.sup.8a or R.sup.8b is as previously described. In a particular embodiment, R.sup.8a and R.sup.8b are H. In another particular embodiment, one of R.sup.8a or R.sup.8b is H, and the other is C.sub.1-4 alkyl. In yet another particular embodiment, one of R.sup.8a or R.sup.8b is H, and the other is C.sub.1-4 alkyl substituted with one OH, C.sub.1-4 alkoxy, or phenyl. In a further particular embodiment, R.sup.8a and R.sup.8b are C.sub.1-4 alkyl. In a more particular embodiment, one of R.sup.8a or R.sup.8b is H, and the other is Me, Et, iPr or tBu. In another more particular embodiment, one of R.sup.8a or R.sup.8b is H, and the other is Me, Et, iPr or tBu, each of which is substituted with one OH, C.sub.1-4 alkoxy, or phenyl. In yet another more particular embodiment, one of R.sup.8a or R.sup.8b is H, and the other is Me, Et, iPr or tBu, each of which is substituted with one OH, OCH.sub.3, or phenyl. In a most particular embodiment, R.sup.4 is NH.sub.2, NHCH.sub.3, N(CH.sub.3).sub.2, NHCH.sub.2Phenyl, or NHCH.sub.2CH.sub.2OCH.sub.3.
[0208] In another embodiment, a compound of the invention is according to Formula I or II, wherein R.sup.1 is 4-7 membered monocyclic heterocycloalkyl comprising 1 to 2 heteroatoms independently selected from N, O, and S. In a particular embodiment, R.sup.1 is azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, tetrahydrofuranyl, tetrahydropyranyl, or dioxanyl. In a more particular embodiment, R.sup.1 is azetidinyl.
[0209] In one embodiment, a compound of the invention is according to Formula I or II, wherein R.sup.1 is 4-7 membered monocyclic heterocycloalkyl comprising 1 to 2 heteroatoms independently selected from N, O, and S, substituted with one or more independently selected C.sub.1-4 alkyl, C(O)C.sub.1-4 alkyl, or C(O)OC.sub.1-4 alkyl. In another embodiment, R.sup.1 is azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, tetrahydrofuranyl, tetrahydropyranyl, or dioxanyl, each of which is substituted with one or more independently selected C.sub.1-4 alkyl, C(O)C.sub.1-4 alkyl, or C(O)OC.sub.1-4 alkyl. In a particular embodiment, R.sup.1 is 4-7 membered monocyclic heterocycloalkyl comprising 1 to 2 heteroatoms independently selected from N, O, and S, substituted with one C.sub.1-4 alkyl, C(O)C.sub.1-4 alkyl, or C(O)OC.sub.1-4 alkyl. In another particular embodiment, R.sup.1 is azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, tetrahydrofuranyl, tetrahydropyranyl, or dioxanyl, each of which is substituted with one C.sub.1-4 alkyl, C(O)C.sub.1-4 alkyl, or C(O)OC.sub.1-4 alkyl. In a more particular embodiment, R.sup.1 is 4-7 membered monocyclic heterocycloalkyl comprising 1 to 2 heteroatoms independently selected from N, O, and S, substituted with one or more independently selected CH.sub.3, C(O)CH.sub.3, or C(O)OC(CH.sub.3).sub.3. In another more particular embodiment, R.sup.1 is azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, tetrahydrofuranyl, tetrahydropyranyl, or dioxanyl, each of which is substituted with one or more independently selected CH.sub.3, C(O)CH.sub.3, C(O)OCH.sub.3, or C(O)OC(CH.sub.3).sub.3. In yet another more particular embodiment, R.sup.1 is azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, tetrahydrofuranyl, tetrahydropyranyl, or dioxanyl, each of which is substituted with one C(O)CH.sub.3, C(O)OCH.sub.3, or C(O)OC(CH.sub.3).sub.3.
[0210] In one embodiment, a compound of the invention is according to Formula I or II, wherein R.sup.1 is phenyl.
[0211] In another embodiment, a compound of the invention is according to Formula I or II, wherein R.sup.1 is phenyl substituted with one or more independently selected R.sup.5 groups. In a particular embodiment, R.sup.1 is phenyl substituted with one, two, or three independently selected R.sup.5 groups. In another particular embodiment, R.sup.1 is phenyl substituted with one R.sup.5 group.
[0212] In one embodiment, a compound of the invention is according to Formula I or II, wherein R.sup.1 is 5-6 membered monocyclic heteroaryl comprising 1 or 2 heteroatoms independently selected from N, O, and S. In a particular embodiment, R.sup.1 is imidazolyl, pyrazolyl, thiazolyl, oxazolyl, pyridinyl, pyrimidinyl or pyrazinyl. In a more particular embodiment, R.sup.1 is pyridinyl.
[0213] In another embodiment, a compound of the invention is according to Formula I or II, wherein R.sup.1 is 5-6 membered monocyclic heteroaryl comprising 1, 2 or 3 heteroatoms independently selected from N, O, and S substituted with one or more independently selected R.sup.5 groups. In another embodiment R.sup.1 is imidazolyl, pyrazolyl, thiazolyl, oxazolyl, pyridinyl, pyrimidinyl or pyrazinyl, each of which is substituted with one or more independently selected R.sup.5 groups. In a particular embodiment, R.sup.1 is 5-6 membered monocyclic heteroaryl comprising 1, 2 or 3 heteroatoms independently selected from N, O, and S substituted with one, two, or three independently selected R.sup.5 groups. In another particular embodiment, R.sup.1 is imidazolyl, pyrazolyl, thiazolyl, oxazolyl, pyridinyl, pyrimidinyl or pyrazinyl, each of which is substituted with one, two, or three independently selected R.sup.5 groups. In a more particular embodiment, R.sup.1 is 5-6 membered monocyclic heteroaryl comprising 1, 2 or 3 heteroatoms independently selected from N, O, and S substituted with one R.sup.5 group. In another more particular embodiment, R.sup.1 is imidazolyl, pyrazolyl, thiazolyl, oxazolyl, pyridinyl, pyrimidinyl or pyrazinyl, each of which is substituted with one R.sup.5 group.
[0214] In one embodiment, a compound of the invention is according to Formula I or II, wherein R.sup.5 is halo, OH, or CN. In a particular embodiment, R.sup.5 is F, Cl, OH, or CN.
[0215] In one embodiment, a compound of the invention is according to Formula I or II, wherein R.sup.5 is C.sub.1-4 alkyl. In a particular embodiment, R.sup.5 is Me, Et, or iPr.
[0216] In another embodiment, a compound of the invention is according to Formula I or II, wherein R.sup.5 is C.sub.1-4 alkyl substituted with one or more independently selected halo, NR.sup.9aR.sup.9b, C(O)NR.sup.9cR.sup.9d, wherein R.sup.9a, R.sup.9b, R.sup.9c, or R.sup.9d is as previously described. In another embodiment, R.sup.5 is Me, or Et, each of which is substituted with one or more independently selected halo, NR.sup.9aR.sup.9b, C(O)NR.sup.9cR.sup.9d. In a particular embodiment, R.sup.5 is C.sub.1-4 alkyl substituted with one, two or three independently selected halo, NR.sup.9aR.sup.9b, or C(O)NR.sup.9cR.sup.9d. In another particular embodiment, R.sup.5 is Me, or Et, each of which is substituted with one, two, or three independently selected halo, NR.sup.9aR.sup.9b, or C(O)NR.sup.9cR.sup.9d. In a more particular embodiment, R.sup.5 is C.sub.1-4 alkyl substituted with one halo, NR.sup.9aR.sup.9b, or C(O)NR.sup.9cR.sup.9d. In another more particular embodiment, R.sup.5 is Me, or Et, each of which is substituted with one halo, NR.sup.9aR.sup.9b, or C(O)NR.sup.9cR.sup.9d. In one embodiment, each R.sup.9a, R.sup.9b, R.sup.9c, or R.sup.9d is independently selected from H, Me, and Et. In a most particular embodiment, R.sup.5 is CF.sub.3, CH.sub.2NH.sub.2, CH.sub.2NHMe, CH.sub.2NMe.sub.2, CH.sub.2C(O)NH.sub.2, CH.sub.2C(O)NHMe, or CH.sub.2C(O)NMe.sub.2.
[0217] In one embodiment, a compound of the invention is according to Formula I or II, wherein R.sup.5 is C.sub.1-4 alkoxy. In a particular embodiment, R.sup.5 is OMe, -OEt, or -OiPr.
[0218] In another embodiment, a compound of the invention is according to Formula I or II, wherein R.sup.5 is C.sub.1-4 alkoxy substituted with one NR.sup.9eR.sup.9f, wherein R.sup.9e are R.sup.9f as previously described. In another embodiment, R.sup.5 is -OEt, substituted with one NR.sup.9eR.sup.9f. In one embodiment, each R.sup.9e, and R.sup.9f, is independently selected from H, Me, and Et. In a most particular embodiment, R.sup.5 is OCH.sub.2CH.sub.2NH.sub.2, OCH.sub.2CH.sub.2NHMe, or OCH.sub.2CH.sub.2NMe.sub.2.
[0219] In another embodiment, a compound of the invention is according to Formula I or II, wherein R.sup.5 is S(O).sub.2C.sub.1-4 alkyl. In a particular embodiment, R.sup.5 is S(O).sub.2CH.sub.3.
[0220] In one embodiment, a compound of the invention is according to Formula IIIa or IIIb:
##STR00011##
wherein R.sup.2, R.sup.3a, R.sup.3b, R.sup.6a, R.sup.6b, the subscript n and Cy are as defined above.
[0221] In one embodiment, a compound of the invention is according to Formula IIIc or IIId:
##STR00012##
wherein R.sup.2, R.sup.3a, R.sup.3b, R.sup.6a, R.sup.6b, the subscript n and Cy are as defined above.
[0222] In one embodiment, a compound of the invention is according to any one of Formulae I-IIId, wherein R.sup.2 is H.
[0223] In one embodiment, a compound of the invention is according to any one of Formulae I-IIId, wherein R.sup.2 is OH.
[0224] In one embodiment, a compound of the invention is according to any one of Formulae I-IIId, wherein R.sup.2 is C.sub.1-4 alkoxy. In a particular embodiment, R.sup.2 is OMe, -OEt, or -OiPr. In a more particular embodiment, R.sup.2 is OMe.
[0225] In one embodiment, a compound of the invention is according to any one of Formulae I-IIId, wherein R.sup.2 is C.sub.1-4 alkyl. In a particular embodiment, R.sup.2 is Me, Et, or iPr. In a more particular embodiment, R.sup.2 is Me, or Et.
[0226] In one embodiment, a compound of the invention is according to any one of Formulae I-IIId, wherein R.sup.2 is C.sub.1-4 alkyl substituted with one OH, or CN. In a particular embodiment, R.sup.2 is Me, or Et, each of which is substituted with one OH, or CN. In a more particular embodiment, R.sup.2 is CH.sub.2OH, or CH.sub.2CN.
[0227] In one embodiment, a compound of the invention is according to any one of Formulae I-IIId, wherein R.sup.2 is C.sub.1-4 alkyl substituted with one C.sub.1-4 alkoxy optionally substituted with one phenyl. In another embodiment, R.sup.2 is Me, or Et, each of which is substituted with one C.sub.1-4 alkoxy optionally substituted with one phenyl. In a particular embodiment, R.sup.2 is C.sub.1-4 alkyl substituted with one OMe, -OEt, each of which is optionally substituted with one phenyl. In another particular embodiment, R.sup.2 is Me, or Et, each of which is substituted with one OMe, -OEt, each of which is optionally substituted with one phenyl. In a more particular embodiment, R.sup.2 is CH.sub.2OCH.sub.3, CH.sub.2OCH.sub.2CH.sub.3, CH.sub.2OCH.sub.2CH.sub.2OCH.sub.3, or CH.sub.2OCH.sub.2Phenyl.
[0228] In one embodiment, a compound of the invention is according to any one of Formulae I-IIId, wherein R.sup.2 is C.sub.1-4 alkyl substituted with one 5-6 membered monocyclic heteroaryl comprising 1 or 2 heteroatoms independently selected from N, O, and S, optionally substituted with one or more independently selected C.sub.10.4 alkyl. In another embodiment, R.sup.2 is Me, or Et, each of which is substituted with one 5-6 membered monocyclic heteroaryl comprising 1 or 2 heteroatoms independently selected from N, O, and S, optionally substituted with one or more independently selected C.sub.1-4 alkyl. In a particular embodiment, R.sup.2 is C.sub.1-4 alkyl substituted with one imidazolyl, pyrrazolyl, oxazolyl, each of which is optionally substituted with one or more independently selected C.sub.1-4 alkyl. In another particular embodiment, R.sup.2 is Me or Et, each of which is substituted with one imidazolyl, pyrrazolyl, oxazolyl, each of which is optionally substituted with one or more independently selected C.sub.1-4 alkyl. In a more particular embodiment, R.sup.2 is C.sub.1-4 alkyl substituted with one imidazolyl, pyrrazolyl, oxazolyl, each of which is optionally substituted with one or more independently selected Me, or Et. In another particular embodiment, R.sup.2 is Me, or Et, each of which is substituted with one imidazolyl, pyrrazolyl, oxazolyl, each of which is optionally substituted with one or more independently selected Me, or Et.
[0229] In one embodiment, a compound of the invention is according to Formula IVa or IVb:
##STR00013##
wherein R.sup.3a, R.sup.3b, R.sup.6a, R.sup.6b, the subscript n and Cy are as defined above.
[0230] In one embodiment, a compound of the invention is according to Formula IVc or IVd:
##STR00014##
wherein R.sup.3a, R.sup.3b, R.sup.6a, R.sup.6b, the subscript n and Cy are as defined above.
[0231] In one embodiment, a compound of the invention is according to any one of Formulae I-IVd, wherein R.sup.3a, and R.sup.3b are both H. In another embodiment, one of R.sup.3a and R.sup.3b is H, and the other is C.sub.1-4 alkyl optionally substituted with one or more halo. In a particular embodiment, one of R.sup.3a and R.sup.3b is H, and the other is Me or Et optionally substituted with one or more halo. In another particular embodiment, one of R.sup.3a and R.sup.3b is H, and the other is Me or Et. In a more particular embodiment, one of R.sup.3a and R.sup.3b is H, and the other is Me, Et or CF.sub.3. In a most particular embodiment, one of R.sup.3a and R.sup.3b is H, and the other is Me. In another most particular embodiment, one of R.sup.3a and R.sup.3b is H, and the other is CF.sub.3. In another most particular embodiment, R.sup.3a and R.sup.3b are both Me.
[0232] In one embodiment, a compound of the invention is according to Formula Va or Vb:
##STR00015##
wherein R.sup.6a, R.sup.6b, the subscript n and Cy are as defined above.
[0233] In one embodiment, a compound of the invention is according to Formula Vc or Vd:
##STR00016##
wherein R.sup.6a, R.sup.6b, the subscript n and Cy are as defined above.
[0234] In one embodiment, a compound of the invention is according to any one of Formulae I-Vd, wherein Cy is 6-10 membered monocyclic or fused bicyclic aryl. In a particular embodiment, Cy is phenyl, or naphthyl. In a more particular embodiment, Cy is phenyl.
[0235] In one embodiment, a compound of the invention is according to any one of Formulae I-Vd, wherein Cy is 5-10 membered monocyclic or fused bicyclic heteroaryl comprising 1, 2 or 3 heteroatoms independently selected from N, O, and S. In a particular embodiment, Cy is pyrazolyl, oxazolyl, oxadiazolyl, thiazolyl, pyridinyl, pyrazinyl, pyridazinyl, pyrimidinyl, indolyl, indazolyl, pyrrolopyridinyl, or benzofuranyl. In a more particular embodiment, Cy is pyridinyl.
[0236] In one embodiment, a compound of the invention is according to Formula VIa, or VIb:
##STR00017##
wherein R.sup.6a, R.sup.6b, and the subscript n are as defined above.
[0237] In one embodiment, a compound of the invention is according to Formula VIc or VId:
##STR00018##
wherein R.sup.6a, R.sup.6b, and the subscript n are as defined above.
[0238] In one embodiment, a compound of the invention is according to any one of Formulae I-VId, wherein R.sup.6a is C.sub.2-4 alkyl optionally substituted with one or more halo. In a particular embodiment, R.sup.6a is CH.sub.2CH.sub.3, or CH(CH.sub.3).sub.2, each of which is optionally substituted with one or more halo. In another particular embodiment, R.sup.6a is C.sub.2-4 alkyl optionally substituted with one or more F. In a more particular embodiment, R.sup.6a is CH.sub.2CH.sub.3, or CH(CH.sub.3).sub.2, each of which is optionally substituted with one or more F. In a most particular embodiment, R.sup.6a is CH.sub.2CH.sub.3, or CH(CH.sub.3).sub.2. In a further most particular embodiment, R.sup.6a is CH.sub.2CH.sub.3.
[0239] In one embodiment, a compound of the invention is according to any one of Formulae I-VId, wherein R.sup.6a is C.sub.1-4 alkoxy optionally substituted with one or more halo. In a particular embodiment, R.sup.6a is OCH.sub.3, OCH.sub.2CH.sub.3, each of which is optionally substituted with one or more halo. In another particular embodiment, R.sup.6a is C.sub.1-4 alkoxy optionally substituted with one or more F. In a more particular embodiment, R.sup.6a is OCH.sub.3, OCH.sub.2CH.sub.3, each of which is optionally substituted with one or more F. In a most particular embodiment, R.sup.6a is OCH.sub.3, OCH.sub.2CH.sub.3, or OCF.sub.3. In a further most particular embodiment, R.sup.6a is OCH.sub.3.
[0240] In one embodiment, a compound of the invention is according to any one of Formulae I-VId, wherein the subscript n is 0.
[0241] In one embodiment, a compound of the invention is according to any one of Formulae I-VId, wherein the subscript n is 1, 2, or 3. In a particular embodiment, the subscript n in 1 or 2.
[0242] In one embodiment, a compound of the invention is according to any one of Formulae I-VId, wherein R.sup.6b is halo, CN, NO.sub.2, C.sub.1-4 alkyl, or C.sub.1-4 alkoxy. In a particular embodiment, R.sup.6b is halo, C.sub.1-4 alkyl, or C.sub.1-4 alkoxy. In a more particular embodiment, R.sup.6b is F, Cl, CH.sub.3, CH.sub.2CH.sub.3, OCH.sub.3, or OCH.sub.2CH.sub.3. In a most particular embodiment, R.sup.6b is F, Cl, CH.sub.3, or OCH.sub.3.
[0243] In one embodiment, a compound of the invention is according to any one of Formulae I-VIb, wherein R.sup.6b is 5-10 membered monocyclic or fused bicyclic heteroaryl comprising 1, 2 or 3 heteroatoms independently selected from N, O, and S, optionally substituted with one or more independently selected halo, C.sub.1-4 alkyl, C.sub.1-4 alkoxy. In another embodiment, R.sup.6b is pyrrazolyl, oxazolyl, oxadiazolyl, thiazolyl, pyridinyl, pyrazinyl, pyridazinyl, or pyrimidinyl, each of which is optionally substituted with one or more independently selected halo, C.sub.1-4 alkyl, C.sub.1-4 alkoxy. In a particular embodiment, R.sup.6b is 5-10 membered monocyclic or fused bicyclic heteroaryl comprising 1, 2 or 3 heteroatoms independently selected from N, O, and S, optionally substituted with one, two, or three independently selected halo, C.sub.1-4 alkyl, or C.sub.1-4 alkoxy. In another particular embodiment, R.sup.6b is pyrrazolyl, oxazolyl, oxadiazolyl, thiazolyl, pyridinyl, pyrazinyl, pyridazinyl, or pyrimidinyl, each of which is optionally substituted with one, two, or three independently selected halo, C.sub.1-4 alkyl, or C.sub.1-4 alkoxy. In a more particular embodiment, R.sup.6b is 5-10 membered monocyclic or fused bicyclic heteroaryl comprising 1, 2 or 3 heteroatoms independently selected from N, O, and S, optionally substituted with one halo, C.sub.1-4 alkyl, C.sub.1-4 alkoxy. In another more particular embodiment, R.sup.6b is pyrrazolyl, oxazolyl, oxadiazolyl, thiazolyl, pyridinyl, pyrazinyl, pyridazinyl, or pyrimidinyl, each of which is optionally substituted with one halo, C.sub.1-4 alkyl, or C.sub.1-4 alkoxy. In a most particular embodiment, R.sup.6b is 5-10 membered monocyclic or fused bicyclic heteroaryl comprising 1, 2 or 3 heteroatoms independently selected from N, O, and S, optionally substituted with one, two, or three independently selected F, Cl, Me, Et, OMe, or -OEt. In another more particular embodiment, R.sup.6b is pyrrazolyl, oxazolyl, oxadiazolyl, thiazolyl, pyridinyl, pyrazinyl, pyridazinyl, or pyrimidinyl, each of which is optionally substituted with one, two, or three independently selected F, Cl, Me, Et, OMe, or -OEt.
[0244] In one embodiment, a compound of the invention is according to any one of Formulae I-VIb, wherein R.sup.6b is NR.sup.9gR.sup.9h, wherein R.sup.9g and R.sup.9h are as previously described. In a particular embodiment, R.sup.9g and R.sup.9h are both H. In another particular embodiment, R.sup.9g and R.sup.9h are both C.sub.1-4 alkyl. In yet another particular embodiment, one of R.sup.9g and R.sup.9h is H, and the other is C.sub.1-4 alkyl. In a more particular embodiment, R.sup.6b is NH.sub.2, NHMe, or NMe.sub.2.
[0245] In one embodiment, a compound of the invention is according to Formula VIIa, or VIIb:
##STR00019##
wherein R.sup.6a, R.sup.6b, and the subscript n are as defined above.
[0246] In one embodiment, a compound of the invention is according to Formula VIIc or VIId:
##STR00020##
wherein R.sup.6a is as defined above, and each R.sup.6b1 and R.sup.6b2 is independently selected from H, halo, CN, NO.sub.2, C.sub.1-4 alkyl, and C.sub.1-4 alkoxy.
[0247] In one embodiment, a compound of the invention is according to Formula VIIa, VIIb, VIIc or VIId wherein R.sup.6a is C.sub.1-4 alkoxy optionally substituted with one or more halo. In a particular embodiment, R.sup.6a is OCH.sub.3, OCH.sub.2CH.sub.3, each of which is optionally substituted with one or more halo. In another particular embodiment, R.sup.6a is C.sub.1-4 alkoxy optionally substituted with one or more F. In a more particular embodiment, R.sup.6a is OCH.sub.3, OCH.sub.2CH.sub.3, each of which is optionally substituted with one or more F. In a most particular embodiment, R.sup.6a is OCH.sub.3, OCH.sub.2CH.sub.3, or OCF.sub.3. In a further most particular embodiment, R.sup.6a is OCH.sub.3.
[0248] In another particular embodiment, a compound of the invention is according to Formula VIIa, VIIb, VIIc or VIId, wherein each R.sup.6b1 and R.sup.6b2 is independently selected from H, halo, CN, NO.sub.2, C.sub.1-4 alkyl, and C.sub.1-4 alkoxy. In a particular embodiment, each R.sup.6b1 and R.sup.6b2 is independently selected from H, F, Cl, CN, NO.sub.2, CH.sub.3, CH.sub.2CH.sub.3, OCH.sub.3, or OCH.sub.2CH.sub.3. In a more particular embodiment, each R.sup.6b1 and R.sup.6b2 is independently selected from H, F, Cl, CH.sub.3, or OCH.sub.3.
[0249] In one embodiment, a compound of the invention is selected from: [0250] 5-[3-[4-(3-methoxyphenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0251] 5-cyclopropyl-5-[3-[4-(3-methoxyphenyl)piperazin-1-yl]-3-oxo-propyl]imidazolidine-2,4-dione, [0252] 5-methyl-5-[3-oxo-3-[4-[4-(trifluoromethoxy)phenyl]piperazin-1-yl]propyl]imidazolidine-2,4-dione, [0253] 5-methyl-5-[3-oxo-3-[4-[2-(trifluoromethoxy)phenyl]piperazin-1-yl]propyl]imidazolidine-2,4-dione, [0254] 5-[3-[4-(4-methoxyphenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0255] 5-[3-[4-(3-methoxyphenyl)piperazin-1-yl]-3-oxo-propyl]-5-(3-pyridyl)imidazolidine-2,4-dione, [0256] 5-[3-[4-(2-methoxyphenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0257] 5-[3-[4-(3-methoxyphenyl)piperazin-1-yl]-3-oxo-propyl]-5-(2-pyridyl)imidazolidine-2,4-dione, [0258] 5-[3-[4-(4-fluoro-3-methoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0259] 5-[3-[4-(4-chloro-3-methoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0260] 5-[3-[4-(3,5-dimethoxyphenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0261] 5-[3-[4-(3-fluoro-5-methoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0262] 5-[3-[4-(3-chloro-5-methoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0263] 5-[3-[4-(3,4-difluoro-5-methoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0264] 5-cyclopropyl-5-[3-[4-(3-fluoro-5-methoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]imidazolidine-2,4-dione, [0265] 5-[3-[4-(3-ethoxyphenyl)piperazin-1l-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0266] 5-cyclopropyl-5-[3-[4-(4-fluoro-3-methoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]imidazolidine-2,4-dione, [0267] 5-cyclopropyl-5-[3-[4-(3,4-difluoro-5-methoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]imidazolidine-2,4-dione, [0268] (R)-5-[3-[4-(3-fluoro-5-methoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione [0269] (R)-5-[3-[4-(3,4-difluoro-5-methoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione [0270] 5-cyclopropyl-5-[3-[(3S)-4-(3,4-difluoro-5-methoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]imidazolidine-2,4-dione, [0271] 5-[3-[(3 S)-4-(3,4-difluoro-5-methoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0272] 5-[3-[4-(3-fluoro-5-methoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]-5-(3-pyridyl)imidazolidine-2,4-dione, [0273] 5-[3-[(3S)-4-(4-chloro-3-methoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0274] 5-[3-[(3 S)-4-(3,5-dimethoxyphenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0275] (5 S)-5-cyclopropyl-5-[(2S)-3-[4-(3-fluoro-5-methoxy-phenyl)piperazin-1-yl]-2-methyl-3-oxo-propyl]imidazolidine-2,4-dione, [0276] (5R)-5-[(2S)-3-[4-(3-fluoro-5-methoxy-phenyl)piperazin-1-yl]-2-methyl-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0277] 5-[3-[4-(3,4-difluoro-5-methoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]-5-(3-pyridyl)imidazolidine-2,4-dione, [0278] 5-cyclopropyl-5-[(2S)-3-[(3S)-4-(3,4-difluoro-5-methoxy-phenyl)-3-methyl-piperazin-1-yl]-2-methyl-3-oxo-propyl]imidazolidine-2,4-dione, [0279] 5-[3-[(3 S)-4-(4-chloro-3,5-dimethoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0280] 5-[3-[4-(4-chloro-3,5-dimethoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0281] 5-[3-[4-(5-methoxy-2-methyl-phenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0282] 5-[3-[4-(3-methoxy-2-methyl-phenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0283] 5-[3-[4-(4-chloro-5-methoxy-2-methyl-phenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0284] 5-[3-[(3S)-4-(5-methoxy-2-methyl-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0285] 5-[3-[(3 S)-4-(4-chloro-5-methoxy-2-methyl-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0286] (5S)-5-cyclopropyl-5-[3-[(3 S)-4-(3,4-difluoro-5-methoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]imidazolidine-2,4-dione, [0287] 5-[3-[4-(4-chloro-3-fluoro-5-methoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0288] 5-[3-[(3S)-4-(4-chloro-3-fluoro-5-methoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0289] 5-[3-[4-(4-chloro-3,5-dimethoxy-phenyl)piperazin-1-yl]-2-methyl-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0290] 5-[3-[4-(4-chloro-3-methoxy-phenyl)-3-methyl-piperazin-1-yl]-2-methyl-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0291] 5-[3-[(3S)-4-(3,4-difluoro-5-methoxy-phenyl)-3-methyl-piperazin-1-yl]-2-methyl-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0292] 5-[3-[4-(3,4-difluoro-5-methoxy-phenyl)piperazin-1-yl]-2-methyl-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0293] 5-[3-[(3S)-4-(4-chloro-3,5-dimethoxy-phenyl)-3-methyl-piperazin-1-yl]-2-methyl-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0294] 5-[3-[(3S)-4-(4-chloro-5-methoxy-2-methyl-phenyl)-3-methyl-piperazin-1-yl]-2-methyl-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0295] 5-[3-[(3S)-4-(3,4-difluoro-5-methoxy-phenyl)-3-methyl-piperazin-1-yl]-2-methyl-3-oxo-propyl]-5-(methoxymethyl)imidazolidine-2,4-dione, [0296] 5-[3-[(3S)-4-(4-chloro-3-methoxy-phenyl)-3-methyl-piperazin-1-yl]-2-methyl-3-oxo-propyl]-5-(methoxymethyl)imidazolidine-2,4-dione, [0297] 5-[3-[4-(3,4-dichloro-5-methoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0298] 5-[3-[(3 S)-4-(3,4-difluoro-5-methoxy-phenyl)-3-methyl-piperazin-1-yl]-2-methyl-3-oxo-propyl]-5-(2-pyridyl)imidazolidine-2,4-dione, [0299] 5-[3-[4-(3-chloro-4-fluoro-5-methoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0300] 5-[3-[4-(3,4-difluoro-5-methoxy-phenyl)piperazin-1-yl]-2-methyl-3-oxo-propyl]imidazolidine-2,4-dione, [0301] 5-[3-[4-(3,4-difluoro-5-methoxy-phenyl)piperazin-1-yl]-2-methyl-3-oxo-propyl]-5-[(3,3-difluoropyrrolidin-1-yl)methyl]imidazolidine-2,4-dione, [0302] 5-[3-[(3 S)-4-(3-chloro-4-fluoro-5-methoxy-phenyl)-3-methyl-piperazin-1l-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0303] (5 S)-5-[(2S)-3-[4-(3,4-difluoro-5-methoxy-phenyl)piperazin-1-yl]-2-methyl-3-oxo-propyl]-5-(2-pyridyl)imidazolidine-2,4-dione, [0304] (5R)-5-[3-[(3 S)-4-(4-chloro-3-methoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0305] (5R)-5-[(2R)-3-[4-(3,4-difluoro-5-methoxy-phenyl)piperazin-1-yl]-2-(hydroxymethyl)-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0306] (5R)-5-[(2S)-3-[4-(3,4-difluoro-5-methoxy-phenyl)piperazin-1-yl]-2-methyl-3-oxo-propyl]-5-ethyl-imidazolidine-2,4-dione, [0307] 5-[3-[4-(4-chloro-2-methoxy-5-methyl-phenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0308] (5 S)-5-[(2S)-3-[4-(3,4-difluoro-5-methoxy-phenyl)piperazin-1-yl]-2-methyl-3-oxo-propyl]-5-(methoxymethyl)imidazolidine-2,4-dione, [0309] 5-cyclopropyl-5-[3-[(3S)-4-(5-methoxy-3-pyridyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]imidazolidine-2,4-dione, [0310] 5-[3-[(3S)-4-(3,4-difluoro-5-methoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-(6-methyl-2-pyridyl)imidazolidine-2,4-dione, [0311] (5S)-5-[3-[(3S)-4-(3-chloro-4-fluoro-5-methoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-cyclopropyl-imidazolidine-2,4-dione, [0312] (5R)-5-[3-[(3 S)-4-(3-chloro-4-fluoro-5-methoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0313] 5-cyclopropyl-5-[3-[(3 S)-4-(5-methoxy-3-pyridyl)-3-methyl-piperazin-1-yl]-2-methyl-3-oxo-propyl]imidazolidine-2,4-dione, [0314] (5S)-5-[3-[(3S)-4-(4-chloro-3-fluoro-5-methoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-cyclopropyl-imidazolidine-2,4-dione, [0315] (5R)-5-[3-[(3S)-4-(4-fluoro-5-methoxy-2-methyl-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0316] (5R)-5-[3-[4-(4-fluoro-5-methoxy-2-methyl-phenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0317] (5R)-5-[3-[4-(3-chloro-2-methoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0318] (5R)-5-[3-[(3 S)-4-(3-chloro-2-methoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0319] (5R)-5-[3-[4-(3-ethylphenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0320] (5R)-5-[3-[4-(4-chloro-3-ethyl-phenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0321] (5R)-5-[3-[4-(3,5-diethylphenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0322] (5R)-5-[3-[4-(4-chloro-2-methoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0323] (5R)-5-[3-[4-(4-fluoro-3,5-dimethoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0324] (5R)-5-[3-[(3 S)-4-(4-chloro-3-ethyl-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0325] (5R)-5-[3-[(3 S)-4-(4-chloro-2-methoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0326] (5R)-5-[3-[4-(5-chloro-2-methoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0327] (5R)-5-[3-[4-(4-fluoro-2-methoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0328] (5R)-5-[3-[4-(4-chloro-3-ethoxy-5-methoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0329] (5R)-5-[3-[(3 S)-4-(4-chloro-3-ethoxy-5-methoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0330] (5R)-5-[3-[(3 S)-4-(4-fluoro-2-methoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0331] (5R)-5-[3-[4-(3-fluoro-2-methoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0332] (5R)-5-[3-[4-(5-fluoro-2-methoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0333] (5R)-5-[3-[(3 S)-4-(3-fluoro-2-methoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0334] (5R)-5-[3-[(3 S)-4-(4-chloro-3-isopropyl-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0335] (5R)-5-[3-[(3S)-4-[4-chloro-3-(trifluoromethoxy)phenyl]-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0336] (5R)-5-[3-[4-(4-chloro-3-isopropyl-phenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0337] (5R)-5-[3-[4-[4-chloro-3-(trifluoromethoxy)phenyl]piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0338] (5R)-5-[3-[(3 S)-4-(4-fluoro-3,5-dimethoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0339] (5R)-5-[3-[(3 S)-4-(3,4-dichloro-5-methoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0340] (5R)-5-[3-[(3 S)-4-(5-chloro-2-methoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0341] (5R)-5-[3-[(3 S)-4-(4,5-dichloro-2-methoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0342] (5R)-5-[3-[(3 S)-4-(4-fluoro-3-methoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0343] (5R)-5-[3-[4-(4,5-dichloro-2-methoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0344] (5R)-5-[3-[(3S)-4-(4-chloro-2-methoxy-5-methyl-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0345] (5R)-5-[3-[(3 S)-4-(3-chloro-5-methoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0346] (5R)-5-[3-[4-(4-chloro-2,5-dimethoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0347] 2-[4-[3-[(3 S)-4-(4-chloro-3-ethyl-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-2,5-dioxo-imidazolidin-4-yl]acetic acid, [0348] (5R)-5-[3-[(3S)-4-[4-chloro-3-(dimethylamino)-5-methoxy-phenyl]-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0349] 5-[3-[(3 S)-4-(4-chloro-3-ethyl-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-(6-methyl-2-pyridyl)imidazolidine-2,4-dione, [0350] 5-[3-[(3 S)-4-[4-chloro-3-(trifluoromethoxy)phenyl]-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-(6-methyl-2-pyridyl)imidazolidine-2,4-dione, [0351] (5R)-5-[3-[4-(4-chloro-5-ethyl-2-methoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0352] (5R)-5-[3-[(3 S)-4-(4-chloro-5-ethyl-2-methoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0353] (5R)-5-[3-[(3 S)-4-(4-chloro-2,5-dimethoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0354] (5R)-5-[3-[(3S)-4-(4-chloro-3-methoxy-5-methyl-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0355] (5R)-5-[3-[(3 S)-4-(4-chloro-3-ethyl-5-methoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0356] (5R)-5-[3-[(3 S)-4-(4-chloro-3-methoxy-phenyl)-3-ethyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0357] (5R)-5-[3-[(3S)-4-[5-methoxy-2-(trifluoromethoxy)phenyl]-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0358] (5R)-5-[3-[(3S)-4-(4-chloro-3-methoxy-phenyl)-3-isopropyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0359] (5R)-5-[3-[(3 S)-4-(4-chloro-3,5-dimethoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0360] (5R)-5-[3-[(3 S)-4-(4-chloro-3-ethoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0361] (5R)-5-[3-[4-(4-chloro-3-methoxy-5-methyl-phenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0362] (5R)-5-[3-[(3 S)-4-(4,5-difluoro-2-methoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0363] tert-butyl 3-[4-[3-[(3S)-4-(4-chloro-3-ethyl-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-2,5-dioxo-imidazolidin-4-yl]propanoate, [0364] (5R)-5-[3-[4-(4-chloro-3-ethyl-5-methoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0365] 3-[4-[3-[(3 S)-4-(4-chloro-3-ethyl-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-2,5-dioxo-imidazolidin-4-yl]propanoic acid, [0366] 5-[3-[(3S)-4-(4-chloro-3-ethyl-5-methoxy-phenyl)-3-methyl-piperazin-1-yl]-2-methyl-3-oxo-propyl]-5-(methoxymethyl)imidazolidine-2,4-dione, [0367] (5R)-5-[3-[4-(4-chloro-3-methoxy-phenyl)-3-(trifluoromethyl)piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0368] (5R)-5-[3-[(3 S)-4-(4-chloro-3-fluoro-5-methoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0369] 5-[3-[(3S)-4-(4-chloro-3-fluoro-5-methoxy-phenyl)-3-methyl-piperazin-1-yl]-2-methyl-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0370] 5-[3-[(3S)-4-(4-chloro-3-fluoro-5-methoxy-phenyl)-3-methyl-piperazin-1-yl]-2-methyl-3-oxo-propyl]-5-(methoxymethyl)imidazolidine-2,4-dione, [0371] (5R)-5-[(2S)-3-[4-(4-chloro-3-fluoro-5-methoxy-phenyl)piperazin-1-yl]-2-methyl-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0372] (5 S)-5-[(2S)-3-[4-(4-chloro-3-fluoro-5-methoxy-phenyl)piperazin-1-yl]-2-methyl-3-oxo-propyl]-5-(methoxymethyl)imidazolidine-2,4-dione, [0373] 5-[3-[(3S)-4-(4-chloro-3-ethyl-phenyl)-3-methyl-piperazin-1-yl]-2-methyl-3-oxo-propyl]-5-(methoxymethyl)imidazolidine-2,4-dione, [0374] 5-[3-[(3S)-4-(4-chloro-3-methoxy-5-methyl-phenyl)-3-methyl-piperazin-1-yl]-2-methyl-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0375] 5-[3-[(3S)-4-(4-chloro-3-methoxy-5-methyl-phenyl)-3-methyl-piperazin-1-yl]-2-methyl-3-oxo-propyl]-5-(methoxymethyl)imidazolidine-2,4-dione, [0376] (5R)-5-[3-[(3R)-4-(3,4-difluoro-5-methoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]imidazolidine-2,4-dione, [0377] (5R)-5-[3-[4-(3,4-difluoro-5-methoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]imidazolidine-2,4-dione, [0378] (5R)-5-[3-[(3S)-4-(3,4-difluoro-5-methoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]imidazolidine-2,4-dione, [0379] 5-cyclopropyl-5-[3-[4-(3-fluoro-5-methoxy-phenyl)piperazin-1-yl]-2-methyl-3-oxo-propyl]imidazolidine-2,4-dione, [0380] 5-[3-[4-(3-fluoro-5-methoxy-phenyl)piperazin-1-yl]-2-methyl-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0381] (5R)-5-[3-[(3 S)-4-(3,5-dimethoxyphenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]imidazolidine-2,4-dione, [0382] (5R)-5-[3-[(3 S)-4-(4-chloro-3-methoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]imidazolidine-2,4-dione, [0383] 5-cyclopropyl-5-[3-[(3S)-4-(3,4-difluoro-5-methoxy-phenyl)-3-methyl-piperazin-1-yl]-2-methyl-3-oxo-propyl]imidazolidine-2,4-dione, [0384] (5R)-5-[3-[4-(4-chloro-3,5-dimethoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]imidazolidine-2,4-dione, [0385] (5R)-5-[3-[4-(5-methoxy-2-methyl-phenyl)piperazin-1-yl]-3-oxo-propyl]imidazolidine-2,4-dione, [0386] (5R)-5-[3-[4-(3-methoxy-2-methyl-phenyl)piperazin-1-yl]-3-oxo-propyl]imidazolidine-2,4-dione, [0387] (5R)-5-[3-[4-(4-chloro-5-methoxy-2-methyl-phenyl)piperazin-1-yl]-3-oxo-propyl]imidazolidine-2,4-dione, [0388] (5R)-5-[3-[(3 S)-4-(4-chloro-3,5-dimethoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]imidazolidine-2,4-dione, [0389] (5R)-5-[3-[(3 S)-4-(5-methoxy-2-methyl-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]imidazolidine-2,4-dione, [0390] (5R)-5-[3-[(3 S)-4-(3-methoxy-2-methyl-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]imidazolidine-2,4-dione, [0391] (5R)-5-[3-[(3S)-4-(4-chloro-5-methoxy-2-methyl-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]imidazolidine-2,4-dione, [0392] (5R)-5-[3-[4-(4-chloro-3-fluoro-5-methoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]imidazolidine-2,4-dione, [0393] (5R)-5-[3-[(3 S)-4-(4-chloro-3-fluoro-5-methoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]imidazolidine-2,4-dione, [0394] (5R)-5-[3-[4-(3,4-dichloro-5-methoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]imidazolidine-2,4-dione, [0395] (5 S)-5-[3-[(3 S)-4-(4-chloro-3-methoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione, [0396] (5R)-5-[3-[4-(3-chloro-4-fluoro-5-methoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]imidazolidine-2,4-dione, [0397] (5R)-5-[3-[(3 S)-4-(3-chloro-4-fluoro-5-methoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]imidazolidine-2,4-dione, [0398] (5R)-5-[3-[4-(4-chloro-3-ethoxy-5-methoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]imidazolidine-2,4-dione, [0399] (5R)-5-[3-[(3 S)-4-(4-chloro-3-ethoxy-5-methoxy-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]imidazolidine-2,4-dione, and [0400] (5R)-5-[3-[4-(4-fluoro-3,5-dimethoxy-phenyl)piperazin-1-yl]-3-oxo-propyl]imidazolidine-2,4-dione,
[0401] In one embodiment, the compound is not (5R)-5-[3-[(3S)-4-(4-chloro-3-methoxy-5-methyl-phenyl)-3-methyl-piperazin-1-yl]-3-oxo-propyl]-5-methyl-imidazolidine-2,4-dione.
[0402] In one embodiment a compound of the invention is not an isotopic variant.
[0403] In one aspect a compound of the invention according to any one of the embodiments herein described is present as the free base.
[0404] In one aspect a compound of the invention according to any one of the embodiments herein described is a pharmaceutically acceptable salt.
[0405] In one aspect a compound of the invention according to any one of the embodiments herein described is a solvate of the compound.
[0406] In one aspect a compound of the invention according to any one of the embodiments herein described is a solvate of a pharmaceutically acceptable salt of a compound.
[0407] While specified groups for each embodiment have generally been listed above separately, a compound of the invention includes one in which several or each embodiment in the above Formula, as well as other formulae presented herein, is selected from one or more of particular members or groups designated respectively, for each variable. Therefore, this invention is intended to include all combinations of such embodiments within its scope.
[0408] While specified groups for each embodiment have generally been listed above separately, a compound of the invention may be one for which one or more variables (for example, R groups) is selected from one or more embodiments according to any of the Formula(e) listed above. Therefore, the present invention is intended to include all combinations of variables from any of the disclosed embodiments within its scope.
[0409] Alternatively, the exclusion of one or more of the specified variables from a group or an embodiment, or combinations thereof is also contemplated by the present invention.
[0410] In certain aspects, the present invention provides prodrugs and derivatives of the compounds according to the formulae above. Prodrugs are derivatives of the compounds of the invention, which have metabolically cleavable groups and become by solvolysis or under physiological conditions the compounds of the invention, which are pharmaceutically active, in vivo. Such examples include, but are not limited to, choline ester derivatives and the like, N-alkylmorpholine esters and the like.
[0411] Other derivatives of the compounds of this invention have activity in both their acid and acid derivative forms, but the acid sensitive form often offers advantages of solubility, tissue compatibility, or delayed release in the mammalian organism (Bundgaard, 1985). Prodrugs include acid derivatives well known to practitioners of the art, such as, for example, esters prepared by reaction of the parent acid with a suitable alcohol, or amides prepared by reaction of the parent acid compound with a substituted or unsubstituted amine, or acid anhydrides, or mixed anhydrides. Simple aliphatic or aromatic esters, amides and anhydrides derived from acidic groups pendant on the compounds of this invention are preferred prodrugs. In some cases it is desirable to prepare double ester type prodrugs such as (acyloxy)alkyl esters or ((alkoxycarbonyl)oxy)alkylesters. Particularly useful are the C.sub.1 to C.sub.8 alkyl, C.sub.2-C.sub.8 alkenyl, aryl, C.sub.7-C.sub.12 substituted aryl, and C.sub.7-C.sub.12 arylalkyl esters of the compounds of the invention.
Pharmaceutical Compositions
[0412] When employed as a pharmaceutical, a compound of the invention is typically administered in the form of a pharmaceutical composition. Such compositions can be prepared in a manner well known in the pharmaceutical art and comprise at least one active compound of the invention according to Formula I. Generally, a compound of the invention is administered in a pharmaceutically effective amount. The amount of compound of the invention actually administered will typically be determined by a physician, in the light of the relevant circumstances, including the condition to be treated, the chosen route of administration, the actual compound of the invention administered, the age, weight, and response of the individual patient, the severity of the patient's symptoms, and the like.
[0413] The pharmaceutical compositions of this invention can be administered by a variety of routes including oral, rectal, transdermal, subcutaneous, intra-articular, intravenous, intramuscular, and intranasal. Depending on the intended route of delivery, a compound of the invention is preferably formulated as either injectable or oral compositions or as salves, as lotions or as patches all for transdermal administration.
[0414] The compositions for oral administration can take the form of bulk liquid solutions or suspensions, or bulk powders. More commonly, however, the compositions are presented in unit dosage forms to facilitate accurate dosing. The term unit dosage forms refers to physically discrete units suitable as unitary dosages for human subjects and other mammals, each unit containing a predetermined quantity of active material calculated to produce the desired therapeutic effect, in association with a suitable pharmaceutical excipient, vehicle or carrier. Typical unit dosage forms include prefilled, premeasured ampules or syringes of the liquid compositions or pills, tablets, capsules or the like in the case of solid compositions. In such compositions, the compound of the invention according to Formula I is usually a minor component (from about 0.1 to about 50% by weight or preferably from about 1 to about 40% by weight) with the remainder being various vehicles or carriers and processing aids helpful for forming the desired dosing form.
[0415] Liquid forms suitable for oral administration may include a suitable aqueous or non-aqueous vehicle with buffers, suspending and dispensing agents, colorants, flavors and the like. Solid forms may include, for example, any of the following ingredients, or compound of the inventions of a similar nature: a binder such as microcrystalline cellulose, gum tragacanth or gelatin; an excipient such as starch or lactose, a disintegrating agent such as alginic acid, Primogel, or corn starch; a lubricant such as magnesium stearate; a glidant such as colloidal silicon dioxide; a sweetening agent such as sucrose or saccharin; or a flavoring agent such as peppermint or orange flavoring.
[0416] Injectable compositions are typically based upon injectable sterile saline or phosphate-buffered saline or other injectable carriers known in the art. As before, the active compound of the invention according to Formula I in such compositions is typically a minor component, often being from about 0.05 to 10% by weight with the remainder being the injectable carrier and the like.
[0417] Transdermal compositions are typically formulated as a topical ointment or cream containing the active ingredient(s), generally in an amount ranging from about 0.01 to about 20% by weight, preferably from about 0.1 to about 20% by weight, preferably from about 0.1 to about 10% by weight, and more preferably from about 0.5 to about 15% by weight. When formulated as an ointment, the active ingredients will typically be combined with either a paraffinic or a water-miscible ointment base.
[0418] Alternatively, the active ingredients may be formulated in a cream with, for example an oil-in-water cream base. Such transdermal formulations are well-known in the art and generally include additional ingredients to enhance the dermal penetration of stability of the active ingredients or the formulation. All such known transdermal formulations and ingredients are included within the scope of this invention.
[0419] A compound of the invention can also be administered by a transdermal device. Accordingly, transdermal administration can be accomplished using a patch either of the reservoir or porous membrane type, or of a solid matrix variety.
[0420] The above-described components for orally administrable, injectable or topically administrable compositions are merely representative. Other materials as well as processing techniques and the like are set forth in Part 8 of Remington's Pharmaceutical Sciences, 17.sup.th edition, 1985, Mack Publishing Company, Easton, Pa., which is incorporated herein by reference.
[0421] A compound of the invention can also be administered in sustained release forms or from sustained release drug delivery systems. A description of representative sustained release materials can be found in Remington's Pharmaceutical Sciences.
[0422] The following formulation examples illustrate representative pharmaceutical compositions that may be prepared in accordance with this invention. The present invention, however, is not limited to the following pharmaceutical compositions.
Formulation 1Tablets
[0423] A compound of the invention according to Formula I may be admixed as a dry powder with a dry gelatin binder in an approximate 1:2 weight ratio. A minor amount of magnesium stearate may be added as a lubricant. The mixture may be formed into 240-270 mg tablets (80-90 mg of active compound of the invention according to Formula I per tablet) in a tablet press.
Formulation 2Capsules
[0424] A compound of the invention according to Formula I may be admixed as a dry powder with a starch diluent in an approximate 1:1 weight ratio. The mixture may be filled into 250 mg capsules (125 mg of active compound of the invention according to Formula I per capsule).
Formulation 3Liquid
[0425] A compound of the invention according to Formula I (125 mg), may be admixed with sucrose (1.75 g) and xanthan gum (4 mg) and the resultant mixture may be blended, passed through a No. 10 mesh U.S. sieve, and then mixed with a previously made solution of microcrystalline cellulose and sodium carboxymethyl cellulose (11:89, 50 mg) in water. Sodium benzoate (10 mg), flavor, and color may be diluted with water and added with stirring. Sufficient water may then be added with stirring. Further sufficient water may be then added to produce a total volume of 5 mL.
Formulation 4Tablets
[0426] A compound of the invention according to Formula I may be admixed as a dry powder with a dry gelatin binder in an approximate 1:2 weight ratio. A minor amount of magnesium stearate may be added as a lubricant. The mixture may be formed into 450-900 mg tablets (150-300 mg of active compound of the invention according to Formula I) in a tablet press.
Formulation 5Injection
[0427] A compound of the invention according to Formula I may be dissolved or suspended in a buffered sterile saline injectable aqueous medium to a concentration of approximately 5 mg/mL.
Formulation 6Topical
[0428] Stearyl alcohol (250 g) and a white petrolatum (250 g) may be melted at about 75 C. and then a mixture of A compound of the invention according to Formula I (50 g) methylparaben (0.25 g), propylparaben (0.15 g), sodium lauryl sulfate (10 g), and propylene glycol (120 g) dissolved in water (about 370 g) may be added and the resulting mixture may be stirred until it congeals.
Methods of Treatment
[0429] In one embodiment, the present invention provides compounds of the invention, or pharmaceutical compositions comprising a compound of the invention, for use in medicine. In a particular embodiment, the present invention provides compounds of the invention or pharmaceutical compositions comprising a compound of the invention, for use in the prophylaxis and/or treatment of inflammatory diseases, and/or diseases involving degradation of cartilage and/or disruption of cartilage homeostasis.
[0430] In another embodiment, the present invention provides compounds of the invention, or pharmaceutical compositions comprising a compound of the invention for use in the manufacture of a medicament for use in the prophylaxis and/or treatment of inflammatory diseases, and/or diseases involving degradation of cartilage and/or disruption of cartilage homeostasis.
[0431] In one embodiment, the present invention provides pharmaceutical compositions comprising a compound of the invention, and another therapeutic agent. In a particular embodiment, the other therapeutic agent is an agent for the prophylaxis and/or treatment of inflammatory diseases, and/or diseases involving degradation of cartilage and/or disruption of cartilage homeostasis.
[0432] In additional method of treatment aspects, this invention provides methods of prophylaxis and/or treatment of inflammatory diseases, and/or diseases involving degradation of cartilage and/or disruption of cartilage homeostasis, which methods comprise the administration of an effective amount of a compound of the invention or one or more of the pharmaceutical compositions herein described for the treatment or prophylaxis of said condition.
[0433] In additional method of treatment aspects, this invention provides methods of prophylaxis and/or treatment of a mammal afflicted with inflammatory diseases, and/or diseases involving degradation of cartilage and/or disruption of cartilage homeostasis, which methods comprise the administration of an effective amount of a compound of the invention or one or more of the pharmaceutical compositions herein described for the treatment or prophylaxis of said condition.
[0434] In one embodiment, the present invention provides compounds of the invention or pharmaceutical compositions comprising a compound of the invention, for use in the prophylaxis and/or treatment of inflammatory diseases. In a particular embodiment, the inflammatory disease is selected from rheumatoid arthritis, and osteoarthritis. More particularly, the inflammatory disease is osteoarthritis.
[0435] In another embodiment, the present invention provides compounds of the invention, or pharmaceutical compositions comprising a compound of the invention for use in the manufacture of a medicament for use in the prophylaxis and/or treatment of inflammatory diseases. In a particular embodiment, the inflammatory disease is selected from rheumatoid arthritis, and osteoarthritis. More particularly, the inflammatory disease is osteoarthritis.
[0436] In additional method of treatment aspects, this invention provides methods of prophylaxis and/or treatment of a mammal afflicted with inflammatory diseases, which methods comprise the administration of an effective amount of a compound of the invention or one or more of the pharmaceutical compositions herein described for the treatment or prophylaxis of said condition. In a particular embodiment, the inflammatory disease is selected from rheumatoid arthritis, and osteoarthritis. More particularly, the inflammatory disease is osteoarthritis.
[0437] In one embodiment, the present invention provides compounds of the invention or pharmaceutical compositions comprising a compound of the invention, for use in the prophylaxis and/or treatment of diseases involving degradation of cartilage and/or disruption of cartilage homeostasis. In a particular embodiment, the diseases involving degradation of cartilage and/or disruption of cartilage homeostasis is selected from osteoarthritis, psoriatic arthritis, juvenile rheumatoid arthritis, gouty arthritis, septic or infectious arthritis, reactive arthritis, reflex sympathetic dystrophy, algodystrophy, achondroplasia, Paget's disease, Tietze syndrome or costal chondritis, fibromyalgia, osteochondritis, neurogenic or neuropathic arthritis, arthropathy, sarcoidosis, amylosis, hydarthrosis, periodical disease, rheumatoid spondylitis, endemic forms of arthritis like osteoarthritis deformans endemica, Mseleni disease and Handigodu disease; degeneration resulting from fibromyalgia, systemic lupus erythematosus, scleroderma and ankylosing spondylitis. More particularly, the diseases involving degradation of cartilage and/or disruption of cartilage homeostasis is osteoarthritis (OA).
[0438] In another embodiment, the present invention provides compounds of the invention, or pharmaceutical compositions comprising a compound of the invention for use in the manufacture of a medicament for use in the prophylaxis and/or treatment of diseases involving degradation of cartilage and/or disruption of cartilage homeostasis. In a particular embodiment, the diseases involving degradation of cartilage and/or disruption of cartilage homeostasis is selected from osteoarthritis, psoriatic arthritis, juvenile rheumatoid arthritis, gouty arthritis, septic or infectious arthritis, reactive arthritis, reflex sympathetic dystrophy, algodystrophy, achondroplasia, Paget's disease, Tietze syndrome or costal chondritis, fibromyalgia, osteochondritis, neurogenic or neuropathic arthritis, arthropathy, sarcoidosis, amylosis, hydarthrosis, periodical disease, rheumatoid spondylitis, endemic forms of arthritis like osteoarthritis deformans endemica, Mseleni disease and Handigodu disease; degeneration resulting from fibromyalgia, systemic lupus erythematosus, scleroderma and ankylosing spondylitis. More particularly, the diseases involving degradation of cartilage and/or disruption of cartilage homeostasis is osteoarthritis (OA).
[0439] In additional method of treatment aspects, this invention provides methods of prophylaxis and/or treatment of a mammal afflicted with diseases involving degradation of cartilage and/or disruption of cartilage homeostasis, which methods comprise the administration of an effective amount of a compound of the invention or one or more of the pharmaceutical compositions herein described for the treatment or prophylaxis of said condition. In a particular embodiment, the diseases involving degradation of cartilage and/or disruption of cartilage homeostasis is selected from osteoarthritis, psoriatic arthritis, juvenile rheumatoid arthritis, gouty arthritis, septic or infectious arthritis, reactive arthritis, reflex sympathetic dystrophy, algodystrophy, achondroplasia, Paget's disease, Tietze syndrome or costal chondritis, fibromyalgia, osteochondritis, neurogenic or neuropathic arthritis, arthropathy, sarcoidosis, amylosis, hydarthrosis, periodical disease, rheumatoid spondylitis, endemic forms of arthritis like osteoarthritis deformans endemica, Mseleni disease and Handigodu disease; degeneration resulting from fibromyalgia, systemic lupus erythematosus, scleroderma and ankylosing spondylitis. More particularly the diseases involving degradation of cartilage and/or disruption of cartilage homeostasis is osteoarthritis (OA).
[0440] Injection dose levels range from about 0.1 mg/kg/h to at least 10 mg/kg/h, all for from about 1 to about 120 h and especially 24 to 96 h. A preloading bolus of from about 0.1 mg/kg to about 10 mg/kg or more may also be administered to achieve adequate steady state levels. The maximum total dose is not expected to exceed about 1 g/day for a 40 to 80 kg human patient.
[0441] For the prophylaxis and/or treatment of long-term conditions, such as degenerative conditions, the regimen for treatment usually stretches over many months or years so oral dosing is preferred for patient convenience and tolerance. With oral dosing, one to four (1-4) regular doses daily, especially one to three (1-3) regular doses daily, typically one to two (1-2) regular doses daily, and most typically one (1) regular dose daily are representative regimens. Alternatively for long lasting effect drugs, with oral dosing, once every other week, once weekly, and once a day are representative regimens. In particular, dosage regimen can be every 1-14 days, more particularly 1-10 days, even more particularly 1-7 days, and most particularly 1-3 days.
[0442] Using these dosing patterns, each dose provides from about 1 to about 1000 mg of a compound of the invention, with particular doses each providing from about 10 to about 500 mg and especially about 30 to about 250 mg.
[0443] Transdermal doses are generally selected to provide similar or lower blood levels than are achieved using injection doses.
[0444] When used to prevent the onset of a condition, a compound of the invention will be administered to a patient at risk for developing the condition, typically on the advice and under the supervision of a physician, at the dosage levels described above. Patients at risk for developing a particular condition generally include those that have a family history of the condition, or those who have been identified by genetic testing or screening to be particularly susceptible to developing the condition.
[0445] A compound of the invention can be administered as the sole active agent or it can be administered in combination with other therapeutic agents, including other compound of the inventions that demonstrate the same or a similar therapeutic activity and that are determined to be safe and efficacious for such combined administration. In a specific embodiment, co-administration of two (or more) agents allows for significantly lower doses of each to be used, thereby reducing the side effects seen.
[0446] In one embodiment, a compound of the invention or a pharmaceutical composition comprising a compound of the invention is administered as a medicament. In a specific embodiment, said pharmaceutical composition additionally comprises a further active ingredient.
[0447] In one embodiment, a compound of the invention is co-administered with another therapeutic agent for the treatment and/or prophylaxis of a disease involving inflammation, particular agents include, but are not limited to, immunoregulatory agents e.g. azathioprine, corticosteroids (e.g. prednisolone or dexamethasone), cyclophosphamide, cyclosporin A, tacrolimus, mycophenolate, mofetil, muromonab-CD3 (OKT3, e.g. Orthocolone), ATG, aspirin, acetaminophen, ibuprofen, naproxen, and piroxicam.
[0448] In one embodiment, a compound of the invention is co-administered with another therapeutic agent for the treatment and/or prophylaxis of arthritis (e.g. rheumatoid arthritis), particular agents include but are not limited to analgesics, non-steroidal anti-inflammatory drugs (NSAIDS), steroids, synthetic DMARDS (for example but without limitation methotrexate, leflunomide, sulfasalazine, Auranofin, sodium aurothiomalate, penicillamine, chloroquine, hydroxychloroquine, azathioprine, tofacitinib, baricitinib, fostamatinib, and cyclosporin), and biological DMARDS (for example but without limitation infliximab, etanercept, adalimumab, rituximab, and abatacept).
[0449] In one embodiment, a compound of the invention is co-administered with another therapeutic agent for the treatment and/or prophylaxis of SLE, particular agents include but are not limited to: human monoclonal antibodies (belimumab (Benlysta)), Disease-modifying antirheumatic drugs (DMARDs) such as antimalarials (e.g. plaquenil, hydroxychloroquine), immunosuppressants (e.g. methotrexate and azathioprine), cyclophosphamide and mycophenolic acid, immunosuppressive drugs and analgesics, such as nonsteroidal anti-inflammatory drugs, opiates (e.g. dextropropoxyphene and co-codamol), opioids (e.g. hydrocodone, oxycodone, MS Contin, or methadone) and the fentanyl duragesic transdermal patch.
[0450] In one embodiment, a compound of the invention is co-administered with another therapeutic agent for the treatment and/or prophylaxis of psoriasis, particular agents include but are not limited to: topical treatments such as bath solutions, moisturizers, medicated creams and ointments containing coal tar, dithranol (anthralin), corticosteroids like desoximetasone (Topicort), fluocinonide, vitamin D3 analogues (for example, calcipotriol), argan oil and retinoids (etretinate, acitretin, tazarotene), systemic treatments such as methotrexate, cyclosporine, retinoids, tioguanine, hydroxyurea, sulfasalazine, mycophenolate mofetil, azathioprine, tacrolimus, fumaric acid esters or biologics such as Amevive, Enbrel, Humira, Remicade, Raptiva and ustekinumab (a IL-12 and IL-23 blocker). Additionally, a compound of the invention may be administered in combination with other therapies including, but not limited to phototherapy, or photochemotherapy (e.g. psoralen and ultraviolet A phototherapy (PUVA)).
[0451] By co-administration is included any means of delivering two or more therapeutic agents to the patient as part of the same treatment regime, as will be apparent to the skilled person. Whilst the two or more agents may be administered simultaneously in a single formulation, i.e. as a single pharmaceutical composition, this is not essential. The agents may be administered in different formulations and at different times.
Chemical Synthetic Procedures
General
[0452] The compound of the invention can be prepared from readily available starting materials using the following general methods and procedures. It will be appreciated that where typical or preferred process conditions (i.e. reaction temperatures, times, mole ratios of reactants, solvents, pressures, etc.) are given, other process conditions can also be used unless otherwise stated. Optimum reaction conditions may vary with the particular reactants or solvent used, but such conditions can be determined by one skilled in the art by routine optimization procedures.
[0453] Additionally, as will be apparent to those skilled in the art, conventional protecting groups may be necessary to prevent certain functional groups from undergoing undesired reactions. The choice of a suitable protecting group for a particular functional group as well as suitable conditions for protection and deprotection are well known in the art [2].
[0454] The following methods are presented with details as to the preparation of a compound of the invention as defined hereinabove and the comparative examples. A compound of the invention may be prepared from known or commercially available starting materials and reagents by one skilled in the art of organic synthesis.
[0455] All reagents are of commercial grade and are used as received without further purification, unless otherwise stated. Commercially available anhydrous solvents are used for reactions conducted under inert atmosphere. Reagent grade solvents are used in all other cases, unless otherwise specified. Column chromatography is performed on silica gel 60 (35-70 m). Thin layer chromatography is carried out using pre-coated silica gel 60F-254 plates (thickness 0.25 mm). .sup.1H NMR spectra are recorded on a 400 MHz Avance Bruker spectrometer or a 300 MHz DPX Bruker spectrometer. Chemical shifts (6) for .sup.1H NMR spectra are reported in parts per million (ppm) relative to tetramethylsilane ( 0.00) or the appropriate residual solvent peak, i.e. CHCl.sub.3 ( 7.27), as internal reference. Multiplicities are given as singlet (s), doublet (d), triplet (t), quartet (q), quintuplet (quin), multiplet (m) and broad (br). Electrospray MS spectra are obtained on a Waters platform LC/MS spectrometer or with Waters Acquity UPLC with Waters Acquity PDA detector and SQD mass spectrometer. Columns used: UPLC BEH C18 1.7 m 2.15 mm VanGuard Pre-column with Acquity UPLC BEH C18 1.7 m 2.130 mm Column or Acquity UPLC BEH C18 1.7 m 2.150 mm Column. All the methods are using MeCN/H.sub.2O gradients. MeCN and H.sub.2O contain either 0.1% Formic Acid or 0.05% NH.sub.3. Preparative LCMS: column used, Waters XBridge Prep C18 5 m ODB 30 mm ID100 mm L (preparative column) and Waters XBridge C18 5 m 4.6 mm ID100 mm L (analytical column). All the methods are using MeCN/H.sub.2O gradients. MeCN and H.sub.2O contain either 0.1% Formic Acid or 0.1% Diethylamine. Chiral HPLC analysis are obtained from a Waters 2690 Alliance HPLC system. Microwave heating is performed with a Biotage Initiator. Optical rotation was determined on a Dr. Kernchen Propol digital automatic polarimeter.
TABLE-US-00001 TABLE I List of abbreviations used in the experimental section: Abbreviation Definition L microliter APMA 4-aminophenylmercuric acetate AUC Area Under the Curve BAL Broncho-alveolar lavage BALF Broncho-alveolar lavage fluid BINAP 2,2-Bis(diphenylphosphino)- 1,1-binaphthalene br. d Broad doublet Boc tert-Butyloxy-carbonyl br. s Broad singlet BSA Bovine serum albumine br. t Broad triplet Cat. Catalytic amount cDNA copy deoxyribonucleic acid Cpd Compound d doublet DavePhos 2-Dicyclohexylphosphino-2- (N,N-dimethylamino)biphenyl DCM Dichloromethane DEAD diethyl azodicarboxylate DIPE Diisopropylether DIPEA N,N-diisopropylethylamine DMA Dimethylacetamide DMAP 4-Dimethylaminopyridine DME Dimethoxyethane DMF N,N-dimethylformamide DMPU 1,3-Dimethyl-3,4,5,6- tetrahydro-2(1H)-pyrimidinone DMSO Dimethylsulfoxide dppf 1,1-Bis(diphenylphosphino) ferrocene EDC 1-ethyl-3-(3- dimethylaminopropyl)carbodiimide) EDCHCl N-(3-Dimethylaminopropyl)- N-ethylcarbodiimide hydrochloride eq. Equivalent Et.sub.2O Diethyl ether EtOAc Ethyl acetate EtOH Ethanol FBS Fetal bovine serum FITC Fluorescein Isothiocyanate g gram h hour HATU O-(7-azabenzotriazol-1-yl)- N,N,N,N-tetramethyluronium hexafluorophosphate HOBt Hydroxybenzotriazole HPLC High pressure liquid chromatography HRP horseradish peroxydase Int Intermediate JohnPhos (2-Biphenyl)di-tert- butylphosphine kg kilogram L liter LCMS Liquid Chromatography-Mass Spectrometry LDA Lithium diisopropylamide LiHMMDS Lithium bis(trimethylsilyl)amide m multiplet m-CPBA 3-Chloroperbenzoic acid MeCN Acetonitrile MEK Methyl ethyl ketone MeOH Methanol mg milligram min minute mL millilitre mmol millimoles MMP Matrix Metallo Proteinase Ms'd Mass measured by LC-MS Mtd Method MW Molecular weight N.A. Not available nBuOH n-Butanol Nva Norvaline NMR Nuclear Magnetic Resonance PBF phosphate buffered formalin PBS Phosphate buffered saline PCR Polymerase chain reaction Pd(PPh.sub.3).sub.4 Tetrakis(triphenylphosphine)palladium (0) Pd/C Palladium on Carbon 10% Pd.sub.2(dba).sub.3 Tris(dibenzylideneacetone) dipalladium(0) PdCl.sub.2dppf [1,1- Bis(diphenylphosphino)ferrocene] dichloropalladium(II) PdCl.sub.2[P(o- Dichlorobis(tri-o- Tol).sub.3].sub.2 tolylphosphine)palladium(II) Pd(OAc).sub.2 Palladium(II) acetate PEG Polyethylene glycol PEPPSI-IPr [1,3-Bis(2,6- Diisopropylphenyl)imidazol-2- ylidene](3- chloropyridyl)palladium(II) dichloride ppm part-per-million q quadruplet QrtPCR quantitative real-time PCR QTL quantitative trait loci r.t. room temperature RNA Ribonucleic acid Rt retention time RuPhos 2-Dicyclohexylphosphino- 2,6-diisopropoxybiphenyl s singlet sept septuplet SFC Supercritical fluid chromatography SM Starting Material Ster Stereochemistry t triplet TBAF Tetra-n-butylammonium fluoride t-BuOH Tert-butanol TBDPSC1 Tert-butyldiphenylsilyl chloride TBSC1 Tert-butyldimethylsilyl chloride TEA Triethylamine TFA Trifluoroacetic acid THF Tetrahydrofuran XantPhos 4,5-Bis(diphenylphosphino)- 9,9-dimethylxanthene XPhos 2-Dicyclohexylphosphino- 2,4,6-triisopropylbiphenyl Bn benzyl BOP; (Benzotriazol-1- yloxy)tris(dimethylamino)phosphonium hexafluorophosphate (COCl).sub.2; Oxalyl chhloride Et.sub.3N Triethyl amine iPrOH; isopropanol NaOtBu; Sodium terbutoxide quin quintuplet MS Mass spectrometry NBS N-Bromosuccinimide Pd(OH).sub.2/C Charcoal supported palladium hydroxide PBu.sub.3 Tri-butyl phosphine PDA; Photodiode Array tBu Tert-Butyl tBuLi Tert-Butyl lithium UPLC Ultra Performance Liquid Chromatography UPLC/MS Ultra Performance Liquid Chromatography coupled mass spectrometry
Example 1. Synthetic Preparation of the Compounds of the Invention
[0456] 1.1. General Synthetic Methods
[0457] 1.1.1.1. Synthetic Methods Overview
[0458] B: Preparation of Ketoester
##STR00021##
[0459] C: Preparation of Ketoamide NH
##STR00022##
[0460] A: Preparation of Arylpiperidine
General methods A: Preparation of arylpiperazine
[0461] Method A1: Buchwald reaction with NBoc-piperazine
[0462] Method A2: HCl NBoc deprotection
[0463] Method A3: HCl NBoc deprotection+basic workup
[0464] Method A4: HCl NBoc deprotection+aqueous workup
[0465] Method A5: Buchwald reaction with NH-piperazine
General methods B: Preparation of ketoester
[0466] Method B1: from Meldrum's acid
[0467] Method B2: esterification
[0468] Method B3: Stetter reaction
[0469] Method B4: via epoxide opening
General method C: preparation of ketoamide
[0470] Method C1: preparation of acrylamide
[0471] Method C2: Stetter reaction
[0472] Method C3: via furan oxidation
General method D: Bucherer Bergs reaction
General method E: Method for preparation of hydantoin propionic acids
General method F: Amide bond formation
[0473] Method F1: EDC/HOBt
[0474] Method F2: HATU
[0475] Method F3: BOP
General method G: Functionalization of final compound
[0476] Method G1: O-debenzylation
[0477] 1.1.1.2. General Methods A: Preparation of Arylpiperazine
[0478] 1.1.1.2.1 Method A1: Buchwald Reaction with NBoc-Piperazine
##STR00023##
[0479] A flask is charged with N-Boc protected piperazine (1 eq.), bromoderivative (0.5-2 eq.), BINAP (0.042-0.12 eq.), NaOtBu (0.7-1.4 eq.) and toluene. The reaction mixture is degassed with N.sub.2 and Pd.sub.2(dba).sub.3 (0.021-0.06 eq.) is added. Reaction mixture is heated at 90-110 C. for 2 h-20 h. The reaction mixture is quenched by addition of water or saturated NaHCO.sub.3 solution, extracted with DCM or EtOAc. The combined organic layers are washed with water and brine, dried (over anhydrous Na.sub.2SO.sub.4 or MgSO.sub.4), filtered and concentrated in vacuo to afford the expected arylpiperazine (used as such or purified by flash chromatography on silica gel).
Illustrative Synthesis of ((S)-4-(4-Chloro-3-methoxy-5-methyl-phenyl)-3-methyl-piperazine-1-carboxylic Acid Tert-Butyl Ester
[0480] ##STR00024##
[0481] A flask is charged with (S)-3-methyl-piperazine-1-carboxylic acid tert-butyl ester (161 mg, 0.800 mmol, 1.2 eq.), 5-Bromo-2-chloro-1-methoxy-3-methyl-benzene (162 mg, 0.690 mmol, 1.0 eq.), BINAP (44 mg, 0.073 mmol, 0.1 eq.), NaOtBu (90 mg, 0.940 mmol, 1.4 eq.) and toluene (1.5 mL). The reaction mixture is degassed with N.sub.2 and Pd.sub.2(dba).sub.3 (31 mg, 0.034 mmol, 0.05 eq.) is added. Reaction mixture is heated at 95 C. for 2 h, quenched with water, extracted with DCM. The combined organic layers are washed with water and brine, dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuo to give crude compound. Crude is used as such. LCMS: MW (calcd): 320; m/z MW (obsd): 321 (M+H).
[0482] 1.1.1.2.2 Method A2 (HCl)
[0483] A flask is charged with N-tert-butoxycarbonyl derivative (1 eq.), HCl 4N in dioxane (10 to 40 eq.) is added. The reaction mixture is stirred at r.t. for 1 h to 2 days. If a precipitate is formed, it is filtered and washed with Et.sub.2O or CH.sub.3CN, otherwise, the reaction mixture is concentrated in vacuo. Both work up afford the expected arylpiperazine as hydrochloride salt.
Illustrative Synthesis of Int 068
[0484] ##STR00025##
[0485] A flask is charged with N-tert-butoxycarbonyl derivative (crude, 0.67 mmol, 1 eq.), HCl 4N in dioxane (4 mL) is added. The reaction mixture is stirred at r.t. overnight and concentrated in vacuo. The residue is triturated in Et.sub.2O, filtered and dried in vacuo to afford the expected product as hydrochloride salt. LCMS: MW (calcd): 255; m/z MW (obsd): 255 (M+H).
[0486] 1.1.1.2.3 Method A3 (HCl+Basic Work Up)
[0487] To a solution of N-tert-butoxycarbonyl derivative (1 eq.) in acetonitrile or DCM is added HCl 4N in dioxane (10 to 40 eq.). The reaction mixture is stirred at r.t. for 1 h to 2 days, concentrated in vacuo and the residue is taken up in water and EtOAc or DCM. The aqueous layer is separated and basified (with either NaOH 1N solution or with a saturated Na.sub.2CO.sub.3 or NaHCO.sub.3 solution) and extracted with EtOAc or DCM. The combined organic layers are dried over anhydrous Na.sub.2SO.sub.4 (or MgSO.sub.4), filtered and concentrated in vacuo to afford the expected arylpiperazine.
Illustrative Synthesis of Int 046
[0488] ##STR00026##
[0489] N-tert-butoxycarbonyl derivative (crude, 0.680 mmol, 1 eq.) in a solution of HCl 4N in dioxane (25 mL) is stirred at r.t. overnight, concentrated in vacuo and the residue is taken up in HCl 1N and DCM. The aqueous layer is separated and basified with NaOH 1N solution or with a saturated Na.sub.2CO.sub.3 solution and extracted with DCM. The combined organic layers are dried over anhydrous Na.sub.2SO.sub.4 (or MgSO.sub.4), filtered and concentrated in vacuo to afford the expected product. LCMS: MW (calcd): 259; m/z MW (obsd): 259-261 (M+H).
[0490] 1.1.1.2.3.1. Method A4 (Aqueous HCl)
Illustrative Synthesis of Int 074
[0491] ##STR00027##
[0492] A flask is charged with with N-tert-butoxycarbonyl derivative (154 mg, 0.45 mmol, 1.0 eq.) and concentrated aqueous HCl (37%, 7.5 mL, 90.0 mmol, 200 eq.). The reaction mixture is stirred for 16 h at r.t. then for 1 h at 80 C. Water (100 mL) is added and the mixture is extracted with EtOAc. The aqueous phase is made basic by the addition of NaOH (3.8 g, 94.5 mmol, 210 eq.) and extracted with DCM. The organic phase is collected, washed with brine, dried over anhydrous MgSO.sub.4, filtered and concentrated in vacuo to afford the expected product. LCMS: MW (calcd): 220; m/z MW (obsd): 221 (M+H).
[0493] 1.1.1.2.4 Method A5: Buchwald Reaction with NH-Piperazine
##STR00028##
[0494] A flask is charged with bromoaryl derivative (1 eq.), piperazine (4-6 eq.), BINAP (0.06-0.22 eq.), NaOtBu (1.4-2.5 eq.) and toluene. The reaction mixture is degassed with N.sub.2 and Pd.sub.2(dba).sub.3 (0.03-0.11 eq.) is added. Reaction mixture is heated at 100-110 C. for 2 h-20 h. The reaction mixture is extracted with HCl 1N solution. The aqueous layer is basified with NaOH 2N solution and extracted with EtOAc or DCM. The combined organic layers are washed with water and brine, dried (over anhydrous Na.sub.2SO.sub.4 or MgSO.sub.4), filtered and concentrated in vacuo to afford the expected arylpiperazine used without further purification.
Illustrative Synthesis of Int 045
[0495] ##STR00029##
[0496] A flask is charged with 5-Bromo-1,2-difluoro-3-methoxy-benzene (400 mg, 1.79 mmol, 1 eq.), piperazine (927 mg, 10.76 mmol, 6 eq.), BINAP (134 mg, 0.215 mmol, 0.12 eq.), NaOtBu (241 mg, 2.511 mmol, 1.4 eq.) and toluene (3 mL). The reaction mixture is degassed with N.sub.2 and Pd.sub.2(dba).sub.3 (99 mg, 0.108 mmol, 0.06 eq.) is added. Reaction mixture is heated at 100 C. overnight. The reaction mixture is extracted with HCl 1N solution. The aqueous layer is basified with NaOH 2N solution and extracted with DCM. The combined organic layers are washed with water and brine, dried over anhydrous MgSO.sub.4, filtered and concentrated in vacuo to afford the expected product. LCMS: MW (calcd): 228. MW (obsd): 229 (M+H).
[0497] 1.1.1.3. General Methods B: Preparation of Ketoester
[0498] 1.1.1.3.1 Method B1:from Meldrum's Acid
##STR00030##
Step i)
[0499] To a solution of the carboxylic acid (1 eq.) in DCM at 0 C. under N.sub.2 atmosphere is added portionwise DMAP (1.5 eq.) then 2,2-Dimethyl-[1,3]dioxane-4,6-dione (1.1 eq.) then EDC.HCl (1.2 eq.). After 10 min at 0 C., the reaction mixture is warmed to r.t. and stirred for 4 h. The reaction mixture is quenched with a solution of KHSO.sub.4 5%. The aqueous phase is extracted with DCM, the combined organic layers are washed with a solution of KHSO.sub.4 5%, water and brine, dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuo. This residue is taken up in anhydrous toluene and benzyl alcohol (1.1 eq.) is added. The reaction mixture is stirred at 120 C. for 16 h to 20 h, concentrated in vacuo and purified by flash chromatography on silica gel to afford the expected -ketoester.
Step ii)
[0500] To a solution of the -ketoester (1 eq.) in MEK are added K.sub.2CO.sub.3 (2 eq.), NaI (0.1 eq.) and bromoderivative (1 eq.). The reaction mixture is stirred at 90 C. for 6 h to 16 h and cooled to r.t. Water is added, reaction mixture acidified to pH 8 and extracted with EtOAc. The combined organic layers are washed with water and brine, dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The residue is purified by flash chromatography on silica gel to afford the expected -ketoester.
Step iii)
[0501] To a solution of the -ketoester (1 eq.) in MeOH are added Pd(OH).sub.2/C (0.01 eq.), and cyclohexene (50 eq.). The reaction mixture is stirred at 70 C. for 19 h. The reaction mixture is filtered on celpure P65 and the filtrate is concentrated in vacuo. The residue is purified by flash chromatography on silica gel to afford the expected -ketoester.
Illustrative Synthesis of Int 029
[0502] ##STR00031##
Step i)
[0503] To a solution of methoxy-acetic acid (5.11 mL, 0.067 mol, 1 eq.) in DCM (160 mL) at 0 C. under N.sub.2 atmosphere is added portionwise DMAP (12.21 g, 0.100 mol, 1.5 eq.) then 2,2-Dimethyl-[1,3]dioxane-4,6-dione (10.56 g, 0.073 mol, 1.1 eq.) then EDC.HCl (15.32 g, 0.080 mol, 1.2 eq.). After 10 min at 0 C., the reaction mixture is warmed to r.t. and stirred for 4 h. The reaction mixture is quenched with a solution of KHSO.sub.4 5%. The aqueous phase is extracted with DCM, the combined organic layers are washed with a solution of KHSO.sub.4 5%, water and brine, dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuo. This residue is taken up in anhydrous toluene (220 mL) and benzyl alcohol (7.59 mL, 0.073 mol, 1.1 eq.) is added. The reaction mixture is stirred at 120 C. for 16 h, concentrated in vacuo and purified by flash chromatography on silica gel (eluting with DCM 100%) to afford the expected 3-ketoester. LCMS: MW (calcd): 222; m/z MW (obsd): 245.3 (M+Na)
Step ii)
[0504] To a solution of the -ketoester (8.96 g, 0.040 mol, 1 eq.) in MEK (120 mL) are added K.sub.2CO.sub.3 (11.14 g, 0.081 mol, 2 eq.), NaI (0.6 g, 0.004 mol, 0.1 eq.) and 2-Bromo-propionic acid tert-butyl ester (6.69 mL, 0.040 mol, 1 eq.). The reaction mixture is stirred at 90 C. for 6 h and cooled to r.t.. Water is added, reaction mixture is acidified to pH 8 and extracted with EtOAc. The combined organic layers are washed with water and brine, dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The residue is purified by flash chromatography on silica gel (eluting with Heptane/EtOAc 100/0 to 50/50) to afford the expected -ketoester. LCMS: MW (calcd): 350; m/z MW (obsd): 373.4 (M+Na)
Step iii)
[0505] To a solution of the -ketoester (6.42 g, 0.018 mol, 1 eq.) in MeOH are added Pd(OH).sub.2/C (0.642 g, 0.002 mol, 0.01 eq.), and cyclohexene (93 mL, 0.916 mol, 50 eq.). The reaction mixture is stirred at 70 C. for 19 h. The reaction mixture is filtered on celpure P65, washed with MeOH and the filtrate is concentrated in vacuo. The residue is purified by flash chromatography on silica gel (eluting with Heptane/EtOAc 100/0 to 70/30) to afford the expected product. LCMS: MW (calcd): 216; m/z MW (obsd): 239.3 (M+Na).
[0506] 1.1.1.3.2 Method B2: Esterification
##STR00032##
[0507] A glass pressure flask is charged with the carboxylic acid (1 eq.), DCM and concentrated H.sub.2SO.sub.4 (0.1 eq.). It is capped and weighted as such. It is then cooled to 45 C., the flask is opened and isobutene is bubbled through the cold reaction mixture for approximatively 5 min. The flask is capped and weighted. The process is repeated until the expected weigh of isobutene is obtained (5 eq.). The reaction mixture is stirred at r.t. for 4 days, then the flask is cooled to 45 C. prior to opening. A saturated NaHCO.sub.3 solution is added portionwise, and the vigorous stirring kept for 30 min. The organic layer is separated; the aqueous layer is extracted with DCM. The combined organic layers are washed with brine, dried over anhydrous MgSO.sub.4 and concentrated in vacuo (with a minimum vacuum of 50 mbar) to afford the expected -ketoester.
Illustrative Synthesis of Int 042
[0508] ##STR00033##
[0509] A glass pressure flask is charged with 2-Methyl-4-oxo-hexanoic acid (Kato et al., 2003) (7.3 g, 50.6 mmol, 1 eq.), DCM (40 mL) and concentrated H.sub.2SO.sub.4 (270 L, 5.06 mmol, 0.1 eq.). The flask is capped and weighted as such. It is then cooled to 45 C., the flask is opened and isobutene is bubbled through the cold reaction mixture for approximatively 5 min. The flask is capped and weighted (11 g of isobutene is condensed). The process is repeated until the expected weigh of isobutene is obtained (14.2 g, 253.2 mmol, 5 eq.). The reaction mixture is stirred at r.t. for 4 days, then the flask is cooled to 45 C. prior to opening. A saturated NaHCO.sub.3 solution is added portionwise, and the vigorous stirring kept for 30 min. The organic layer is separated; the aqueous layer is extracted with DCM. The combined organic layers are washed with brine, dried over anhydrous MgSO.sub.4 and concentrated in vacuo (with a minimum vacuum of 50 mbar) to afford the expected product.
[0510] 1.1.1.3.3 Method B3: Stetter Reaction
##STR00034##
[0511] A vial is charged with aldehyde (1 eq.), tert-butyl ester acrylate (1 eq.), PBu.sub.3 (1 eq.) and dry THF. The vial is capped and heated at 70 C. for 2 h to 16 h. The reaction mixture is partitioned between EtOAc and water. The combined organic layers are washed with brine, dried over anhydrous MgSO.sub.4, filtered and concentrated in vacuo to afford the expected -ketoester after purification by flash chromatography on silica gel.
Illustrative Synthesis of Int 014
[0512] ##STR00035##
[0513] To a solution of Pyridine-3-carbaldehyde (16 mL, 170.29 mmol, 1.0 eq.) in THF (300 mL) is added PBu.sub.3 (42.537 mL, 170.29 mmol, 1.0 eq.) and the reaction mixture is heated at 50 C. for 5 min. tert-butyl ester acrylate (24.803 mL, 170.29 mmol, 1 eq.) is added and the reaction mixture is stirred at 80 C. for 3 h. This process (heating 3 h and addition of tert-butyl ester acrylate) is repeated until no evolution is observed by TLC (EtOAc) and UPLC/MS. The reaction mixture is concentrated in vacuo and the residue is purified by flash chromatography on silica gel (eluting with Heptane/EtOAc 100/0 to 0/100) to afford the expected product. LCMS: MW (calcd): 235; m/z MW (obsd): 236 (M+H).
[0514] 1.1.1.3.4 Method B4: Via Epoxide Opening
##STR00036##
Step i)
[0515] To a solution of alkene (1 eq.) in DCM at 0 C., is added m-CPBA (1.5 eq.) and the reaction mixture is stirred at r.t. overnight. The white precipitate is filtered and washed with DCM. The filtrate is washed with a saturated NaHCO.sub.3 solution, brine, dried over anhydrous MgSO.sub.4 and concentrated in vacuo. The residue is purified by flash chromatography on silica gel to afford the expected epoxide.
Step ii)
[0516] A sealed tube is charged with the epoxide (1 eq.), EtOH and secondary amine (1.5 eq.). After heating at reflux for 3 h30, the reaction mixture is concentrated in vacuo. The residue is taken up in DCM, washed with a saturated NH.sub.4Cl solution, dried over anhydrous MgSO.sub.4, filtered and concentrated in vacuo to afford the expected aminoalcohol used in next step without further purification.
Step iii)
[0517] A two necked flask, under N.sub.2 atmosphere, is charged with dry DCM and (COCl).sub.2 (1.1 eq.). The reaction mixture is cooled to 70 C., a solution of DMSO (2.4 eq.) in dry DCM is added dropwise and the reaction mixture is stirred at 70 C./60 C. for 45 min. A solution of the aminoalcohol (1 eq.) in dry DCM is added dropwise and the reaction mixture is stirred for 1 h at 60 C. Et.sub.3N (5 eq.) is added dropwise. Reaction mixture stirred at 40 C. for 30 min then warmed to r.t. and stirred overnight. Water is added, the organic layer is separated and washed with brine, dried over anhydrous MgSO.sub.4 and concentrated in vacuo. The residue is purified by flash chromatography on silica gel to afford the expected -ketoester.
Illustrative Synthesis of Int 021
[0518] ##STR00037##
Step i)
[0519] To a solution of Int 022 (2 g, 11.8 mmol, 1 eq.) in DCM (20 mL) at 0 C., is added m-CPBA (3.05 g, 17.7 mmol, 1.5 eq.) and the reaction mixture is stirred at r.t. overnight. The white precipitate is filtered and washed with DCM. The filtrate is washed with a saturated NaHCO.sub.3 solution, brine, dried over anhydrous MgSO.sub.4 and concentrated in vacuo. The residue is purified by flash chromatography on silica gel (eluting with Heptane/EtOAc 100/0 to 80/20) to afford the expected epoxide.
Step ii)
[0520] A sealed tube is charged with the epoxide (0.9 g, 4.84 mmol, 1 eq.), EtOH (15 mL) and 3,3-difluoropyrrolidine hydrochloride (0.903 g, 6.29 mmol, 1.3 eq.). After heating at reflux for 4 h30, the reaction mixture is concentrated in vacuo. The residue is taken up in DCM, washed with a saturated NH.sub.4Cl solution, dried over anhydrous MgSO.sub.4, filtered and concentrated in vacuo. The residue is purified by flash chromatography on silica gel (eluting with DCM/acetone 100/0 to 90/10) to afford the expected product.
Step iii)
[0521] A two necked flask, under N.sub.2 atmosphere, is charged with dry DCM (5 mL) and (COCl).sub.2 (0.153 mL, 1.81 mmol, 1.1 eq.). The reaction mixture is cooled to 70 C., a solution of DMSO (0.281 mL, 3.96 mmol, 2.4 eq.) in dry DCM (0.5 mL) is added dropwise and the reaction mixture is stirred at 70 C./60 C. for 45 min. A solution of the aminoalcohol (0.450 g, 1.65 mmol, 1 eq.) in dry DCM (2 mL) is added dropwise and the reaction mixture is stirred for 1 h at 60 C. Et.sub.3N (1.19 mL, 8.24 mmol, 5 eq.) is added dropwise. Reaction mixture stirred at 40 C. for 30 min then warmed to r.t. and stirred overnight. Water is added, the organic layer is separated and washed with brine, dried over anhydrous MgSO.sub.4 and concentrated in vacuo. The residue is purified by flash chromatography on silica gel (eluting with DCM/acteone 90/10) to afford the expected product.
[0522] 1.1.1.4. General Method C: Preparation of Ketoamide
[0523] 1.1.1.4.1 Method C1: Preparation of Acrylamide
##STR00038##
[0524] To a solution of piperazine (1 eq.) and Et.sub.3N (1.5 eq.) in DCM at 0 C. is added dropwise the acryloyl chloride derivative (1.5 eq.). Reaction mixture is stirred at 0 C. for 1 h and allowed to reach r.t.. Water and DCM are added, the organic layer is separated. The aqueous layer is extracted with DCM, the combined organic layers are washed with brine and dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuo to afford the expected acrylamide after purification by flash chromatography on silica gel.
Illustrative Synthesis of Int 001
[0525] ##STR00039##
[0526] To a solution of 1-(3-Fluoro-5-methoxy-phenyl)-piperazine (2.06 g, 9.8 mmol, 1 eq.) and Et.sub.3N (1.5 mL, 14.7 mmol, 1.5 eq.) in DCM at 0 C. is added dropwise 2-Methyl-acryloyl chloride (2.05 mL, 14.7 mmol, 1.5 eq.). Reaction mixture is stirred at 0 C. for 1 h and allowed to reach r.t.. Water and DCM are added, the organic layer is separated. The aqueous layer is extracted with DCM, the combined organic layers are washed with brine and dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The residue is purified by flash chromatography on silica gel (eluting with DCM/MeOH 100/0 to 90/10) to afford the expected product. LCMS: MW (calcd): 278; m/z MW (obsd): 279 (M+H).
[0527] 1.1.1.4.2 Method C2: Stetter Reaction
##STR00040##
[0528] A vial is charged with bis(1,5-cyclooctadiene)rhodium(I) tetrafluoroborate (0.10 eq.), 1,4-bis(diphenylphosphino)butane (0.10 eq.), dry DCM and sealed with a septum. The flask is evacuated and refilled with H.sub.2 (3 times) and the reaction mixture is stirred under an atmosphere of H.sub.2. After 3 h, volatiles are removed under a nitrogen stream. The residue is combined with acrylamide (1 eq.), aldehyde (1.5 equiv.) and 1,2-dichloroethane in a vial under a N.sub.2 atmosphere. The vial is sealed with a cap and heated at 100 C. After 16 h, the mixture is concentrated in vacuo and purified by flash chromatography on silica gel to afford the expected -ketoamide.
Illustrative Synthesis of Int 007
[0529] ##STR00041##
[0530] A vial is charged with bis(1,5-cyclooctadiene)rhodium(I) tetrafluoroborate (0.054 g, 0.132 mmol, 0.10 eq.), 1,4-bis(diphenylphosphino)butane (0.056 g, 0.132 mmol, 0.10 eq.), dry DCM (2 mL) and sealed with a septum. The flask is evacuated and refilled with H.sub.2 (3 times) and the reaction mixture is stirred under an atmosphere of H.sub.2. After 3 h, volatiles are removed under a nitrogen stream. The residue is combined with Int 001 (0.397 g, 1.328 mmol, 1 eq.), cyclopropane carboxaldehyde (0.406 g, 2.00 mmol, 1.5 equiv.) and 1,2-dichloroethane (2 mL) in a vial under a N2 atmosphere. The vial is sealed with a cap and heated at 100 C. After 2 days, the mixture is concentrated in vacuo. The residue is purified by flash chromatography on silica gel (eluting with Heptane/EtOAc 100/0 to 0/100, then DCM/MeOH 90/10) to afford Int 007. LCMS: MW (calcd): 348; m/z MW (obsd): 349 (M+H).
[0531] 1.1.1.4.3 Method C3: Via Furan Oxidation
##STR00042##
Step i)
[0532] To a solution of phosphonate (1.1 eq.) in EtOH is added K.sub.2CO.sub.3 (1.2 eq.). The reaction mixture is stirred at r.t. for 2 h prior to addition of the aldehyde (1 eq.). The reaction mixture is stirred at r.t. (1 h to 3 h), diluted with EtOAc and filtered on celpure P65. The filtrate is concentrated in vacuo. The residue is taken up in EtOAc and washed with a saturated NH.sub.4Cl solution, a saturated NaHCO.sub.3 solution, brine and dried over anhydrous MgSO.sub.4, filtered, concentrated in vacuo and purified by flash chromatography on silica gel to afford the expected ,-unsaturated ketone.
Step ii)
[0533] To a solution of the ,-unsaturated ketone (1 eq.) in dry MeOH are added PdCl.sub.2 (0.1 eq.) and 2-methylfuran (2 eq.). The reaction mixture is stirred at r.t. for 3 h to 24 h, diluted with EtOAc and filtered on celpure P65. The filtrate is concentrated in vacuo and purified by flash chromatography on silica gel to afford the expected ketone.
Step iii)
[0534] To a solution of ketone (1 eq.) in Heptane/EtOAc/water (1/3/4) is added NaIO.sub.4 (7 eq.). The reaction mixture is stirred for 10 min then RuCl.sub.3.3H.sub.2O (0.02 eq.) is added. The reaction mixture is stirred for 30 min to 1 h30, filtered on celpure P65, washed with MeCN and the filtrate is concentrated in vacuo. The residue is purified by flash chromatography on silica gel to afford the expected -ketoacid.
Illustrative Synthesis of Int 018
[0535] ##STR00043##
Step i)
[0536] To a solution of dimethyl acetylmethylphosphonate (14.22 g, 73.24 mmol, 1.1 eq.) in EtOH (150 mL) is added K.sub.2CO.sub.3 (11 g, 79.90 mmol, 1.2 eq.). The reaction mixture is stirred at r.t. for 2 h prior to addition of benzyloxy-acetaldehyde (10 g, 66.59 mmol, 1 eq.). The reaction mixture is stirred at r.t. for 3 h, diluted with EtOAc and filtered on celpure P65. The filtrate is concentrated in vacuo. The residue is taken up in EtOAc and washed with a saturated NH.sub.4Cl solution, a saturated NaHCO.sub.3 solution, brine and dried over anhydrous MgSO.sub.4, filtered, concentrated in vacuo and purified by flash chromatography on silica gel (eluting with Heptane/EtOAc 100/0 to 80/20) to afford the expected ,-unsaturated ketone.
Step ii)
[0537] To a solution of the ,-unsaturated ketone (8.7 g, 45.73 mmol, 1 eq.) in dry MeOH (183 mL) are added PdCl.sub.2 (0.811 g, 0.457 mmol, 0.1 eq.) and 2-methylfuran (8.25 mL, 91.46 mmol, 2 eq.). The reaction mixture is stirred at r.t. for 3 h, diluted with EtOAc and filtered on celpure P65. The filtrate is concentrated in vacuo and purified by flash chromatography on silica gel eluting with Heptane/EtOAc 100/0 to 85/15) to afford the expected ketone.
Step iii)
[0538] To a solution of ketone (1 g, 3.67 mmol, 1 eq.) in Heptane/EtOAc/water (6 mL/18 mL/24 mL) is added NaIO.sub.4 (5.48 g, 25.69 mmol, 7 eq.). The reaction mixture is stirred for 10 min then RuCl.sub.3.3H.sub.2O (0.019 g, 0.073 mmol, 0.02 eq.) is added. The reaction mixture is stirred for 1 h15, filtered on celpure P65, washed with MeCN and the filtrate is concentrated in vacuo. The residue is purified by flash chromatography on silica gel (eluting with DCM/MeOH 98/2 to 95/5) to afford the expected product (stored at 4 C.).
[0539] 1.1.1.5. General Method D: Bucherer Bergs Reaction
##STR00044##
G.sub.7=O-Alk.sub.1, Alk.sub.2-N-Alk.sub.3
[0540] A pressure reactor or an open round bottom flask equipped with a condenser is charged with a solution of (NH.sub.4).sub.2CO.sub.3 or (NH.sub.4)HCO.sub.3 (8-12 eq.) in water. KCN (2 to 4 eq.) is added portionwise then a solution of -ketoester or -ketoamide (1 eq.) in EtOH is added. The vessel is sealed and heated at 60-90 C. for 1 h to 2 days. The reaction mixture is cooled to r.t., combined with water and extracted with AcOEt or CHCl.sub.3/nBuOH 10%. The combined organic layers are washed with water and brine, dried (over anhydrous Na.sub.2SO.sub.4 or MgSO.sub.4), filtered and concentrated in vacuo. The residue is either recrystallized or purified by flash chromatography on silica gel to afford the expected hydantoin derivative.
Illustrative Synthesis of (R)-5-Methyl-5-((S)-2-methyl-3-oxo-butyl)-imidazolidine-2,4-dione+(S)-5-Methyl-5-((R)-2-methyl-3-oxo-butyl)-imidazolidine-2,4-dione
[0541] ##STR00045##
[0542] A pressure reactor is charged with a solution of (NH.sub.4).sub.2CO.sub.3 (79.4 g, 0.826 mol, 8 eq.) in water (400 mL). KCN (20 g, 0.307 mol, 3 eq.) is added portionwise then a solution of -ketoester (19.15 g, 0.103 mol, 1 eq.) in EtOH (400 mL) is added. The vessel is sealed and heated at 90 C. overnight. The reaction mixture is cooled to r.t., combined with water and extracted with CHCl.sub.3/nBuOH 10%. The combined organic layers are washed with brine, dried over anhydrous MgSO.sub.4, filtered, concentrated in vacuo.
[0543] The above reaction is performed twice and the two crude residues are gathered for recrystallization. A flask is charged with the two crude residues, EtOH (250 mL) is added and the reaction mixture is heated at reflux. Upon complete dissolution, the reaction mixture is allowed to cool to r.t. for 2 days, it is filtered and the crystalline solid is combined with EtOH (200 mL), heated to reflux, cooled to r.t. overnight and filtered to afford the expected hydantoin as a trans-Me racemic mixture (LCMS: >99% de, MW (calcd): 256; m/z MW (obsd): 257 (M+H)).
Illustrative Synthesis of Cpd 02
[0544] ##STR00046##
[0545] A pressure reactor is charged with (NH.sub.4).sub.2CO.sub.3 (0.645 g, 6.71 mmol, 10 eq.), KCN (0.175 g, 2.69 mmol, 4 eq.), Int 003 (0.248 g, 0.671 mmol, 1 eq.), EtOH (4 mL) and water (2 mL). The vessel is sealed and heated at 60 C. for 40 h. The reaction mixture is cooled to r.t., combined with water and extracted with DCM. The combined organic layers are washed with brine, dried over anhydrous MgSO.sub.4, filtered, concentrated in vacuo. Purification by flash chromatography on silica gel (eluting with DCM/iPrOH 20/1). (LCMS: MW (calcd): 386; m/z MW (obsd): 387 (M+H)).
[0546] 1.1.1.6. General Method E: Method for Preparation of Hydantoin Propionic Acids
##STR00047##
[0547] A flask is charged with tert-butyl ester (1 eq.) and HCl 4N in dioxane (5 to 40 eq.). In some cases, an additional solvent such as DCM, dioxane or water is added to increase solubility. The reaction mixture is stirred at r.t. for 1 h to 4 days until complete conversion. The reaction mixture is either concentrated in vacuo or filtered and washed with Et.sub.2O to afford the expected carboxylic acid.
Illustrative Synthesis of Int 040
[0548] ##STR00048##
[0549] A flask is charged with Int 041 (3.6 g, 13.32 mmol, 1 eq.) and HCl 4N in dioxane (33.3 mL, 133 mmol, 10 eq.). The reaction mixture is stirred at r.t. for 2 days and concentrated in vacuo to afford the expected product.
[0550] 1.1.1.7. General Method F: Amide Bond Formation
##STR00049##
[0551] 1.1.1.7.1 Method F1: EDC/HOBt
[0552] A solution of acid (1 eq.), Et.sub.3N (3 to 4 eq.), HOBt (0.1 to 1.1 eq.) in DMF (or DCM) is stirred at r.t.. EDC.HCl (1 to 1.2 eq.) is added, then amine (0.95 to 2 eq.) is added and the reaction mixture is stirred at r.t. for 5 h to 2 days. The reaction mixture is partitioned between DCM (or EtOAC) and water, extracted with DCM (or EtOAc). The combined organic layers are washed with water and brine, dried over anhydrous Na.sub.2SO.sub.4 (or MgSO.sub.4), filtered, concentrated in vacuo and purified by flash chromatography on silica gel or preparative LCMS to afford the expected amide.
Illustrative Synthesis of Cpd 06
[0553] ##STR00050##
[0554] A solution of 3-(2,5-Dioxo-4-pyridin-3-yl-imidazolidin-4-yl)-propionic acid (100 mg, 0.40 mmol, 1 eq.), Et.sub.3N (168 L, 1.20 mmol, 3 eq.), HOBt (57 mg, 0.42 mmol, 1.05 eq.) in DMF (1 mL) is stirred at r.t.. EDC.HCl (81 mg, 0.42 mmol, 1.05 eq.) is added, then 1-(3-Methoxy-phenyl)-piperazine (116 mg, 0.60 mmol, 1.5 eq.) is added and the reaction mixture is stirred at r.t. overnight. The reaction mixture is partitioned between DCM and water, extracted with DCM. The combined organic layers are washed with water and brine, dried over anhydrous MgSO.sub.4, filtered, concentrated in vacuo and purified by preparative LCMS to afford the expected product. LCMS: MW (calcd): 423; m/z MW (obsd): 424 (M+H).
[0555] 1.1.1.7.2 Method F2: HATU
[0556] A flask is charged with acid (1 eq.), amine (0.85 to 1.1 eq.), HATU (0.85 to 1.1 eq.) and DMF (or THF). DIPEA (2 to 6 eq.) is added and the reaction mixture is stirred at r.t. for 5 h to 2 days. The reaction mixture is partitioned between EtOAc and water, extracted with EtOAc. The combined organic layers are washed with water and brine, dried (over anhydrous Na.sub.2SO.sub.4, MgSO.sub.4, or hydrophobic column), filtered, concentrated in vacuo and purified by flash chromatography on silica gel or preparative LCMS to afford the expected amide.
Illustrative Synthesis of Cpd 105
[0557] ##STR00051##
[0558] A flask is charged with Int 043 (70 mg, 0.35 mmol, 1.1 eq.), Int 068 (95 mg, 0.32 mmol, 1 eq.), HATU (127 mg, 0.34 mmol, 1.05 eq) and DMF (3 mL). DIPEA (167 L, 0.96 mmol, 3 eq.) is added and the reaction mixture is stirred at r.t. overnight. The reaction mixture is partitioned between EtOAc and water, extracted with EtOAc. The combined organic layers are washed with water and brine, dried over hydrophobic column, filtered, concentrated in vacuo and purified by flash chromatography on silica gel (eluting with DCM/MeOH 100/0 to 96/4) to afford the expected product. LCMS: MW (calcd): 517; m/z MW (obsd): 423-425 (M+H).
[0559] 1.1.1.7.3 Method F3: BOP
[0560] A flask is charged with acid (1 eq.), DMF (or DCM), DIPEA or Et.sub.3N (2 to 6 eq.) and BOP (0.77 to 1.1 eq.). After 5-15 min, amine (0.77 to 1.5 eq.) is added and the reaction mixture is stirred at r.t. for 5 h to 2 days. The reaction mixture is partitioned between EtOAc (or DCM) and water, extracted with EtOAc (or DCM). The combined organic layers are washed with water and brine, dried (over anhydrous Na.sub.2SO.sub.4, MgSO.sub.4, or hydrophobic column), filtered, concentrated in vacuo and purified by flash chromatography on silica gel or preparative LCMS to afford the expected amide.
Illustrative Synthesis of Cpd005
[0561] ##STR00052##
[0562] A flask is charged with 3-(4-Methyl-2,5-dioxo-imidazolidin-4-yl)-propionic acid (50 mg, 0.27 mmol, 1 eq.), DMF (2 mL), DIPEA (0.142 mL, 0.81 mmol, 3 eq.) and BOP (119 mg, 0.27 mmol, 1.0 eq.). After 5-15 min, 1-(4-methoxyphenyl)piperazine (79 mg, 0.41 mmol, 1.5 eq.) is added and the reaction mixture is stirred at r.t. overnight. The reaction mixture is partitioned between DCM and water, extracted with DCM. The combined organic layers are washed with water and brine, concentrated in vacuo and purified by flash chromatography on silica gel (eluting with DCM/EtOAc 90/10) afford the expected product. LCMS: MW (calcd): 360; m/z MW (obsd): 361 (M+H).
[0563] 1.1.1.8. General Method G: Functionalization of Final Compound
[0564] 1.1.1.8.1 Method G1: O-Debenzylation
##STR00053##
[0565] To a solution of benzyloxy derivative (1 eq.) in dry THF or MeOH under argon atmosphere is added Pd(OH).sub.2/C (50% w/w). The reaction mixture is stirred under H.sub.2 atmosphere at r.t. for 5 h to 2 days then filtered on celpure P65. The filtrate is concentrated in vacuo and purified by flash chromatography on silica gel to afford the expected alcohol.
Illustrative Synthesis of Cpd 56
[0566] ##STR00054##
[0567] To a solution of Int 005 (105 mg, 0.20 mmol, 1 eq.) in dry THF (10 mL) under argon atmosphere is added Pd(OH).sub.2/C (50 mg, 50% w/w). The reaction mixture is stirred under H.sub.2 atmosphere at r.t. for 5 h then filtered on celpure P65. The filtrate is concentrated in vacuo and purified by flash chromatography on silica gel (eluting with DCM/MeOH 100/0 to 90/10) to afford the expected product. LCMS: MW (calcd): 426; m/z MW (obsd): 427 (M+H).
[0568] 1.2. Preparation of the Compounds of the Invention.
[0569] 1.2.1. Cpd 98
##STR00055##
[0570] A flask is charged with Int 009 (28 mg, 0.06 mmol, 1.0 eq.) and a solution of HCl in dioxane (4N) (1 mL) is added, and stirring is kept at room temperature for 3 h. Reaction mixture is diluted with water, solution of NaHCO.sub.3 is added and extracted with DCM. Organic layers are combined and evaporated under reduced pressure to obtain crude product which is purified by flash chromatography on silica gel (DCM/MeOH 100/0 to 92/8) to afford the expected carboxylic acid. LCMS: MW (calcd): 450; m/z MW (obsd): 451-453 (M+H).
[0571] 1.2.2. Cpd 116
##STR00056##
[0572] A flask is charged with Cpd 114 (68 mg, 0.013 mmol, 1.0 eq.) and a solution of HCl in dioxane (4.0M, 10 mL, 40 mmol, 300 eq.). The flask is capped with an oil bubbler and slowly flushed with a stream of N.sub.2. After 64 h, volatiles are removed via rotary evaporation, and the residue is dissolved in a solution of HCl in dioxane (4.0M, 10 mL, 40 mmol, 300 eq.). The reaction mixture is allowed to stir at r.t. for 40 h. Volatiles are removed via rotary evaporation. The residue is dissolved in DMSO and purified by preparative LC-MS to afford the expected product. LCMS: MW (calcd): 464; m/z MW (obsd): 465 (M+H).
[0573] 1.2.3. Int 002
##STR00057##
Step i)
[0574] A vial is charged with 1,6-dioxaspiro[4.4]nonane-2,7-dione (47.4 mg, 0.30 mmol, 1 eq), Int 062 (79 mg, 0.29 mmol, 0.95 eq), dry dioxane (2 mL), and triethyl amine (0.2 mL, 1.4 mmol, 4.7 eq). After 16 h, the mixture is combined with DCM (100 mL) and aqueous H.sub.3PO.sub.4/NaH.sub.2PO.sub.4 (1M, 100 mL) in a seperatory funnel and agitated. The organic phase is collected, washed with brine (100 mL), and dried over MgSO.sub.4. After filtration, volatiles are removed via rotary evaporation to give the expected product which is used in the following synthetic step without further purification.
Step ii)
[0575] A pressure vessel is charged with the acid synthesized in step i) (0.92 mol) and DCM (10 mL), and cooled in a NaCl/ice bath (20 C.). Isobutene (3.06 g, 54.5 mmol, 59 eq) is condensed into the cold solution, and concentrated H.sub.2SO.sub.4 (0.1 mL, 1.8 mmol, 2.0 eq) is added. The vessel is hermetically sealed, and then the cold bath is removed. After 16 h, the vessel is cooled in a NaCl/ice bath (20 C.), and opened. Et.sub.3N (1.0 mL, 7.2 mmol, 7.8 eq) is added, and the cold bath is removed. Once all volatiles had evaporated, the mixture is combined with H.sub.2O (100 mL) and DCM (100 mL) in a separatory funnel, and agitated. The organic phase is collected, washed with brine (100 mL) and dried over MgSO.sub.4. After filtration, volatiles are removed from the filtrate via rotary evaporation. The residue is purified by flash chromatography on silica gel (EtOAc/DCM 1:4), to afford the expected compound Int 002.
[0576] 1.2.4. Int 010
##STR00058##
[0577] A solution of n-Butyl lithium (1.6M in hexane) (25 mL, 40 mmol, 2.0 eq) is added at 0 C. to a stirred solution of 1,1,1,3,3,3-hexamethyldisilazane (8.5 mL, 41 mmol, 2.04 eq) in anhydrous THF (17 mL). After cooling to 78 C., tertbutyl acetate (5.44 mL, 40 mmol, 2.0 eq) is added within 20 min to the solution and stirring is continued for 45 min. The resulting o-lithio acetic ester solution is added dropwise over 30 min to a solution of succinic anhydride (2 g, 20 mmol, 1.0 eq) in THF (24 mL). The resulting mixture is stirred for 3 h in a methanol/dry ice bath while the temperature is allowed to increase to 20 C.
[0578] The reaction mixture is warmed up to room temperature, then concentrated HCl (4 mL) and water (25 mL) are added. The organic solvent is evaporated, and the resulting aqueous solution is adjusted to pH=2, and extraction with ethyl acetate followed. Organic layers are combined, dried over Na.sub.2SO.sub.4, filtered, and concentrated under reduced pressure to give the expected product (used in the next step without further purification).
[0579] 1.2.5. Int 017
##STR00059##
[0580] To a solution of Int 018 (530 mg, 2.24 mmol, 1 eq.) in toluene (7 mL) is added N,N-dimethylformamide di-tert-butyl acetal (2.69 mL, 11.2 mmol, 5 eq.). Reaction mixture is heated at 100 C. in a sealed tube for 4.5 h, quenched by addition of a saturated NaHCO.sub.3 solution at 0 C., extracted with EtOAc. The combined organic layers are washed with saturated NaHCO.sub.3 solution, brine, dried over anhydrous Na.sub.2SO.sub.4, filtered, concentrated in vacuo and purified by flash chromatography on silica gel (Heptane/EtOAc 100/0 to 60/40) to afford the expected product. LCMS: MW (calcd): 292; m/z MW (obsd): 315 (M+Na)
[0581] 1.2.6. Int 025
##STR00060##
[0582] A three neck flask is charged with a solution of alkene Int 022 (6.3 g, 37 mmol, 1 eq.) and suddan III (cat.) in DCM and cooled at 78 C. O.sub.3 is bubbled trough the reaction mixture until the color became deep blue. The reaction mixture is purged with N.sub.2 for 30 min, Me.sub.2S is added and the reaction mixture is allowed to warm to r.t. overnight. The reaction mixture is washed with water and brine, dried over anhydrous MgSO.sub.4, filtered and concentrated in vacuo. Purification by flash chromatography on silica gel (Heptane/EtOAc 100/0 to 80/20) affords the expected product.
[0583] 1.2.7. Int 026
##STR00061##
Step i)
[0584] To a solution of Meldrum's acid (2,2-dimethyl-[1,3]dioxane-4,6-dione, 50.10 g, 0.347 mol, 1 eq.) in DCM (500 mL) and pyridine (90 mL, 1.11 mol, 3.2 eq.) at 0 C., cyclopropanecarbonyl chloride (35 mL, 0.386 mol, 1.1 eq.) is added dropwise. After 2 h, the cold bath is removed and the reaction mixture is stirred at r.t. overnight and combined with a solution of HCl 2N. The organic layer is collected, washed with brine, dried over anhydrous MgSO.sub.4, filtered over activated charcoal and concentrated in vacuo. This residue is taken up in ethanol (300 mL) and stirred at reflux overnight, concentrated in vacuo and purified by flash chromatography on silica gel (Heptane/EtOAc 80/20) to afford the expected -ketoester. LCMS: MW (calcd): 156; m/z MW (obsd): 157 (M+H); 179 (M+Na)
Step ii)
[0585] To a solution of the -ketoester (16.09 g, 0.103 mol, 1 eq.) in MEK (200 mL) are added K.sub.2CO.sub.3 (28.56 g, 0.207 mol, 2 eq.), NaI (1.65 g, 0.011 mol, 0.1 eq.) and 2-Bromo-propionic acid tert-butyl ester (18 mL, 0.108 mol, 1.04 eq.). The reaction mixture is heated at reflux for 40 h and cooled to r.t.. Water is added, reaction mixture acidified to pH 8 and extracted with EtOAc. The combined organic layers are washed with water and brine, dried over anhydrous MgSO.sub.4, filtered and concentrated in vacuo to afford the expected -ketoester used as such in next step. LCMS: MW (calcd): 284; m/z MW (obsd): 307 (M+Na)
Step iii)
[0586] To a solution of the -ketoester (29.2 g, 0.103 mol, 1 eq.) in EtOH (100 mL) is added a solution of NaOH (12.6 g, 0.315 mol, 3 eq.) in water (100 mL). The reaction mixture is heated at reflux for 16 h, cooled to r.t., diluted with water (500 mL) and cooled in an ice bath. To this is added dropwise H.sub.3PO.sub.4 (85%, 4 mL, 0.059 mol) and conc. HCl (24 mL, 0.288 mol), the ice bath is removed and reaction mixture is stirred at r.t. for 30 min. The reaction mixture is cooled in an ice bath and a solution of NaOH (17 g, 0.425 mol) in water (50 mL) is added to adjust the pH to 8. The solution is combined with DCM, the aqueous layer is collected, cooled in an ice bath and the pH adjusted to pH=2 with conc. HCl. The solution is saturated with NaCl and extracted with DCM. The combined organic layers are dried over anhydrous MgSO.sub.4, filtered, concentrated in vacuo to afford the expected product. LCMS: MW (calcd): 156; m/z MW (obsd): 157 (M+H); 179 (M+Na).
[0587] 1.2.8. Int 033
##STR00062##
Step i)
[0588] A solution of LDA (3.0 L, 5.98 mol, 1.17 eq.) in THF (2.5 L) is cooled to 78 C. A solution of 1-cyclopropylethanone (460 g, 5.11 mol, 1 eq.) in THF (0.5 L) is added dropwise, then warmed to 20 C. and stirred for 30 min. The reaction mixture is cooled to 78 C. and tert-butyl bromoacetate (997 g, 5.11 mol, 1 eq.) in THF (0.5 L) is added slowly. The reaction is stirred at 0 C. overnight, quenched with saturated NH.sub.4Cl aq. (3.3 L), extracted with EtOAc (0.5 L3), washed with water (0.5 L2), saturated NH.sub.4Cl aq. (1 L), and brine (1 L), dried over anhydrous Na.sub.2SO.sub.4. Purification by distillation under reduced pressure (5 mbar, 95 C.) affords the expected -ketoester.
Step ii)
[0589] A mixture of -ketoester 4-Cyclopropyl-4-oxo-butyric acid tert-butyl ester (120 g, 605 mmol, 1 eq.), (NH.sub.4).sub.2CO.sub.3 (494 g, 5.15 mol, 8.5 eq.), NaCN (60 g, 1.45 mol, 2.4 eq.), H.sub.2O (600 mL) and ethanol (600 mL) is heated at 60 C. for 18 h in the sealed reactor. The reaction mixture is poured in a mixture of EtOAc (900 mL) and water (900 mL), and the aqueous layer is additionally extracted with EtOAc (3600 mL). The organic layer is concentrated until only about 100 mL EtOAc left, and added 500 mL petroleum ether dropwise to afford the expected hydantoin derivative.
Step iii)
[0590] A flask is charged with a solution of hydantoin (200 g, 746 mmol, 1.0 eq.) in dioxane (100 mL) and is cooled in an ice bath, HCl 4N in dioxane (1 L) is added slowly. The reaction mixture is stirred at r.t. for 4 h and concentrated in vacuo. The resulting solid is suspended in 240 mL of acetonitrile, then stirred at reflux for 1 h, and allowed to cool down to r.t. under stirring. The resulting solid is separated by filtration, washed twice with acetonitrile (230 mL), and finally dried under vacuum at 45 C. to afford the expected carboxylic acid. LC/MS: MW (calcd): 212; m/z MW (obsd): 211 (MH).
[0591] 1.2.9. Int 034
##STR00063##
[0592] The racemic hydantoin propionic acid is separated by SFC to afford a fast eluting isomer ((R)-enantiomer) and a slow eluting isomer ((S)-enantiomer).
[0593] The purification is done in 2 stages.
[0594] Conditions of the first separation: preparative SFC, Column: ChiralPak AD-10 m, 30050 mm I.D., Mobile phase: A for CO.sub.2 and B for Ethanol, Gradient: B 45%, Flow rate: 200 mL/min, Back pressure: 100 bar, Column temperature: 38 C., Wavelength: 220 nm, Cycletime: 10.0 min. The compound is dissolved in methanol to 120 mg/mL, and loaded on the column (16 mL per injection). After separation, the fractions are dried off via rotary evaporator to get the desired isomers.
[0595] Conditions of the second separation: Prep HPLC, Column: C18, 25050 mm I.D., Mobile phase: A for H.sub.2O and B for Acetonitrile, Gradient: B 5%-20% in 15 min linearly, Flow rate: 80 mL/min, Wavelength: 220 nm. The compound is dissolved in methanol (100 mg/mL) and loaded on the column (10 mL per injection). After separation, the fraction is concentrated via rotary evaporator and the remaining aqueous layer is lyophilized.
[0596] 1.2.10. Int 043 and 138
##STR00064##
[0597] The racemic 3-(4-Methyl-2,5-dioxo-imidazolidin-4-yl)propionic acid (805 g) is separated by SFC to afford 384 g of the faster eluting isomer and 388 g of the slower eluting isomer. Conditions of the separation: Instrument: Thar350 preparative SFC, Column: ChiralPak AD-10m, 30050 mm I.D., Mobile phase: A for CO.sub.2 and B for IPA (0.1% TFA), Gradient: B 25%, Flow rate: 220 mL/min, Back pressure: 100 bar, Column temperature: 38 C., Wavelength: 210 nm, Cycletime: -3.8 min, Sample preparation: Compound is dissolved in methanol to 80 mg/mL, Injection: 1.0 mL per injection, Work up: After separation, the fractions are dried off via rotary evaporator at bath temperature 40 C. to get the desired isomers.
[0598] 1.2.11. Int 117
##STR00065##
[0599] 5-Bromo-2-fluoro-4-methyl-phenol (1.50 g, 7.3 mmol, 1.0 eq.), potassium carbonate (3.03 g, 22.0 mmol, 3.0 eq.) and methyl iodide (1.37 mL, 22.0 mmol, 3.0 eq.) are dissolved in acetonitrile (12 mL) and submitted to microwave irradiation for 20 min at 100 C. The reaction mixture is evaporated under reduced pressure, dissolved in water and extracted with DCM. The combined organic layers are dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuo to afford the expected product.
[0600] 1.2.12. Int 118
##STR00066##
[0601] 1-Bromo-4-chloro-2,5-difluoro-benzene (1.0 g, 4.4 mmol, 1.0 eq.) is dissolved in MeOH (10 mL) and a solution of sodium methoxide in MeOH (25% wt, 1.90 mL, 8.8 mmol, 2.0 eq.) is added. The reaction mixture is submitted to microwave irradiation for 50 min at 120 C. The reaction mixture is diluted with water and extracted with DCM. The combined organic layers are dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuo. Crystallization upon standing and filtration afford the expected product.
[0602] 1.2.13. Int 119
##STR00067##
[0603] 1-Bromo-4-chloro-2-fluoro-5-methyl-benzene (1.0 g, 4.5 mmol, 1.0 eq.) is dissolved in MeOH (10 mL) and a solution of sodium methoxide in MeOH (25% wt, 1.9 mL, 9.0 mmol, 2.0 eq.) is added. The reaction mixture is submitted to microwave irradiation for 50 min at 120 C. The reaction mixture is diluted with water and extracted with DCM. The combined organic layers are dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuo. Crystallization upon standing and filtration afford the expected product.
[0604] 1.2.14. Int 120
##STR00068##
[0605] A solution of 1-bromo-4-chloro-3,5-difluorobenzene (100 mg, 0.44 mmol, 1.0 eq.) and sodium methylate (59 mg, 1.1 mmol, 2.5 eq.) in DMA (0.6 mL) is heated at 100 C. for 3 h. Sodium methoxide (12 mg, 0.22 mmol, 0.5 eq.) is added and the mixture is heated at 100 C. for 1 h, then cooled to r.t. Water (6 mL) is added and the solid is collected by filtration and dried under suction to give the expected product.
[0606] 1.2.15. Int 121
##STR00069##
[0607] A solution of 1-bromo-4-chloro-3,5-difluorobenzene (300 mg, 1.3 mmol, 1.0 eq.) and sodium methoxide (105 mg, 2.6 mmol, 2.0 eq.) in MeOH (5.2 mL) is heated at 70 C. for 18 h. The methanol is evaporated under vacuo, and the residue is partitioned between water and DCM. The aqueous layer is extracted with DCM. The combined organic layers are washed with brine, dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuo to afford the expected product which is used as such in the next step.
[0608] 1.2.16. Int 122
##STR00070##
[0609] 1-bromo-3,4-dichloro-5-fluorobenzene (365 mg, 1.5 mmol, 1.0 eq.) is heated in a 0.5M solution of sodium methoxide in methanol (6.0 mL, 3.0 mmol, 2.0 eq.) at 70 C. for 4 h, then at 85 C. for 18 h. The reaction mixture is cooled in an ice water bath. The solid is filtered and dried under suction to afford the expected product.
[0610] 1.2.17. Int 123
##STR00071##
[0611] A solution of 1-bromo-3,4-difluoro-5-methoxy-benzene (1.11 g, 5.0 mmol, 1.0 eq.) and sodium methoxide (405 mg, 7.5 mmol, 1.5 eq.) in DMA (5 mL) is heated at 100 C. for 1 h. The reaction mixture is then poured into 60 mL of water and ice, stirred and the resulting solid is filtered and dried under suction to afford the expected product.
[0612] 1.2.18. Int 124
##STR00072##
[0613] A solution of 1-bromo-4-chloro-3-fluoro-5-methoxy-benzene (542 mg, 2.3 mmol, 1.0 eq.) and sodium ethoxide (308 mg, 4.6 mmol, 2.0 eq.) in DMA (2.3 mL) is heated at 110 C. for 1.5 h. The reaction mixture is then poured into 100 mL of water. The aqueous layer is extracted 3 times with EtOAc. The combined organic phases are washed successively with water and brine, dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuo to afford the expected product which is used as such in the next step.
[0614] 1.2.19. Int 125
##STR00073##
Step i)
[0615] 1-bromo-4-chloro-3,5-difluorobenzene (682 mg, 3.0 mmol, 1.0 eq.), dimethylamine hydrochloride (734 mg, 9.0 mmol, 3.0 eq.) and DIPEA (2.1 mL, 12.0 mmol, 4.0 eq.) are heated in DMA (2.1 mL) in a sealed microwave vial at 125 C. for 18 h. The reaction mixture is then poured into water and brine. The aqueous layer is extracted 3 times with EtOAc. The combined organic phases are washed successively with water and brine, dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuo to afford the expected product. LCMS: MW (calcd): 251; m/z MW (obsd): 252-254 (M+H).
Step ii)
[0616] A solution of (5-bromo-2-chloro-3-fluoro-phenyl)-dimethyl-amine (599 mg, 2.4 mmol, 1.0 eq.) and sodium methoxide (259 mg, 4.8 mmol, 2.0 eq.) in DMA (2.4 mL) is heated at 110 C. for 2 h. The reaction mixture is then poured into 1000 mL of water and ice, stirred and extracted 3 times with EtOAc. The combined organic phases are washed successively with water and brine, dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuo to afford the expected product. LCMS: MW (calcd): 263; m/z MW (obsd): 264-266 (M+H).
[0617] 1.2.20. Int 126
##STR00074##
Step i)
[0618] To a solution of 2-methoxy-6-methylaniline (13.47 g, 98.3 mmol, 1.0 eq.) in DCM (1.3 L) and MeOH (520 mL) is added benzyltrimethylammonium tribromide (38.3 g, 98.3 mmol, 1.0 eq.) and calcium carbonate (39.3 g, 393.2 mmol, 4.0 eq.). The reaction mixture is stirred at r.t. for 1 h. The solid is filtered and washed with DCM. The filtrate is washed twice with water. The combined aqueous layers are extracted twice with Et.sub.2O. The combined organic phases are dried over anhydrous Na.sub.2SO.sub.4, filtered, concentrated in vacuo to afford the expected product. LCMS: MW (calcd): 215; m/z MW (obsd): 216-218 (M+H).
Step ii)
[0619] To 4-bromo-2-methoxy-6-methyl-aniline (20.85 g, 96.5 mmol, 1.0 eq.) in a 6N aqueous HCl solution (54 mL, 326 mmol, 3.4 eq.) at 0 C. is added slowly, under vigorous stirring, a 0 C.-cooled solution of sodium nitrite (6.49 g, 94 mmol, 1.0 eq.) in water (27 mL). The resulting mixture is added quickly to a 0 C.-cooled solution of CuCl (46.1 g, 465.5 mmol, 4.8 eq.) in concentrated HCl (41 mL), and the flask is rinsed with 75 mL of water. The reaction mixture is stirred at 0 C. for 30 min, then at reflux for 18 h. The reaction mixture is cooled back to r.t., diluted with water and brine and extracted 3 times with DCM. The combined organic phases are washed successively with water and brine, dried over anhydrous Na.sub.2SO.sub.4, filtered, concentrated in vacuo. The residue is stirred in 300 mL of heptane at 100 C. for 15 min, hot filtered through Whatman glass microfibers filter and washed with heptane. The filtrate is concentrated under reduced pressure and the residue is purified by flash chromatography on silica gel to afford the expected product.
[0620] 1.2.21. Int 127
##STR00075##
[0621] A mixture of 5-bromo-2-chlorophenol (300 mg, 1.5 mmol, 1.0 eq.), ethyl iodide (233 L, 2.9 mmol, 2.0 eq.) and potassium carbonate (400 mg, 2.9 mmol, 2.0 eq.) in acetone (3 mL) is heated at 55 C. in a sealed vial for 18 h. Water is added and the reaction mixture is extracted twice with EtOAc. The combined organic phases are washed with water, dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuo to afford the expected product.
[0622] 1.2.22. Int 128
##STR00076##
Step i)
[0623] N-boc-2-methoxy aniline (2.0 g, 9.0 mmol, 1.0 eq) is stirred in dry Et.sub.2O at 78 C. t-BuLi in pentane (1.6 M, 17 mL, 27 mmol, 3.0 eq) is then added dropwise at 78 C. Once addition is complete, the mixture is stirred at 78 C. for 10 min, then 20 min at 0 C. The mixture is then cooled back to 78 C., and a solution of iodoethane (1.4 mL, 18.0 mmol, 2.0 eq) in dry Et.sub.2O is then added dropwise, and stirring is kept at 78 C. After a few min, the dry ice/acetone bath is removed, and the mixture is warm up to room temperature, and left at rt. overnight. The mixture is then carefully quenched with water. Brine is added, and the aqueous layer is extracted with Et.sub.2O (three times). The combined organic layers are dried over Na.sub.2SO.sub.4, and concentrated under reduced pressure and the residue is purified by flash chromatography (Heptane/AcOEt 100/0 to 85/15), to afford the expected compound. LCMS: MW (calcd): 251; m/z MW (obsd): 252 (M+H).
Step ii)
[0624] N-boc-substituted aniline (945 mg, 3.76 mmol, 1.0 eq) is stirred in dioxane (5 mL), and HCl (4N in dioxane) is added. The mixture is stirred at rt; for 1 h30, and water and EtOAc are added. NaOH (2M) is added to the aqueous layer until a pH higher than 10 is reached and is then extracted with DCM (3 times). The combined organic layers are washed with water, brine, dried over Na.sub.2SO.sub.4, filtered, and concentrated under reduced pressure to afford the expected deprotected aniline.
Step iii) and Step iv)
[0625] Identical procedures from Int 126 Step i) and step ii) were applied for step iii) and step iv), to afford Int 128.
[0626] 1.2.23. Int 129
##STR00077##
[0627] A mixture of 3-chloro-4-fluoroanisole (2.52 g, 15.7 mmol, 1.0 eq.), silver trifluoroacetate (8.53 g, 38.6 mmol, 2.5 eq.), iodine (7.94 g, 31.3 mmol, 2.0 eq.) and chloroform (100 mL) is stirred at r.t. for 16 h. The reaction mixture is filtered through a silica plug on a fritted funnel and rinsed with chloroform. The filtrate is washed successively with an aqueous solution of Na.sub.2S.sub.2O.sub.3, water and brine, dried over MgSO.sub.4, filtered and concentrated in vacuo. The residue is purified by flash chromatography on silica gel to afford the expected product.
[0628] 1.2.24. Int 130
##STR00078##
[0629] A mixture of 4-chloro-3-fluoroanisole (2.52 g, 15.7 mmol, 1.0 eq.), silver trifluoroacetate (8.57 g, 38.8 mmol, 2.5 eq.), iodine (7.95 g, 31.3 mmol, 2.0 eq.) and chloroform (100 mL) is stirred at r.t. for 16 h. The reaction mixture is filtered through a silica plug on a fritted funnel and rinsed with chloroform. The filtrate is washed successively with an aqueous solution of Na.sub.2S.sub.2O.sub.3, water and brine, dried over MgSO.sub.4, filtered and concentrated in vacuo. The residue is purified by flash chromatography on silica gel to afford the expected product.
[0630] 1.2.25. Int 131
##STR00079##
[0631] 5-Bromo-3-chloro-2-fluorophenol (113 mg, 0.5 mmol, 1.0 eq.) is dissolved in MeCN (1 mL) in a microwave vial. Iodomethane (94 L, 1.5 mmol, 3.0 eq.) and potassium carbonate (138 mg, 1.0 mmol, 2.0 eq.) are successively added. The vial is submitted to microwave irradiation at 100 C. for 10 min. The mixture is filtered and the salts are washed with EtOAc. The filtrate is then partitioned between water and EtOAc. The organic phase is dried over Na.sub.2SO.sub.4, filtered and concentrated in vacuo to afford the expected product, which is used as such in the next step.
[0632] 1.2.26. Int 132
##STR00080##
[0633] To 2-chloro-4-methoxytoluene (344 L, 2.5 mmol, 1.0 eq.) in MeCN (1.5 mL) at 0 C. is added NBS (500 mg, 3.0 mmol, 1.2 eq.) in 2 portions (2nd half after 30 min stirring). The reaction mixture is allowed to stir at r.t. for 18 h. The solvent is removed in vacuo, and the residue is dissolved in DCM and washed with brine, dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuo to afford the expected product as a mixture, which is used as such in the next step.
[0634] 1.2.27. Int 133
##STR00081##
Step i)
[0635] To a solution of 1-bromo-2-chloro-4-methoxybenzene (2.0 g, 9.0 mmol, 1.0 eq.) in dioxane (180 mL) are added a diethylzinc solution in THF (18 mL, 1M, 18.0 mmol, 2.0 eq.) and [1,1-bis(diphenylphosphino)ferrocene]dichloropalladium (II) (330 mg, 0.45 mmol, 0.05 eq.). The resulting solution is bubbled through with argon and stirred at reflux for 18 h. The reaction mixture is cooled to r.t., diluted with water (100 mL) and extracted 3 times with DCM. The combined organic phases are dried over anhydrous Na.sub.2SO.sub.4, filtered, concentrated in vacuo and purified by flash chromatography on silica gel to afford the expected product.
Step ii)
[0636] To 2-chloro-1-ethyl-4-methoxybenzene (1.36 g, 8.0 mmol, 1.0 eq.) in polyethyleneglycol (8 mL) cooled at 0 C. is added NBS (1.49 g, 8.4 mmol, 1.05 eq.) portionwise. The reaction mixture is allowed to warm to r.t. and is stirred for 18 h. The reaction mixture is diluted with water (20 mL) and extracted 3 times with DCM. The combined organic phases are dried over anhydrous Na.sub.2SO.sub.4, filtered, concentrated in vacuo and purified by flash chromatography on silica gel to afford the expected product.
[0637] 1.2.28. Int 134
##STR00082##
[0638] Int 134 is prepared from 2-Amino-pentanedioic acid using same experimental procedure as for preparation of Int 136
[0639] 1.2.29. Int 136
##STR00083##
[0640] A round bottomed flask (1 L) is charged with (R)-2-Amino-pentanedioic acid, H.sub.2O and and heated at 80 C. After 6 h, the flask is allowed to cool to room temperature. Aqueous HCl (6 N, 200 mL, 1.2 mol) is slowly added. The flask was then heated at 60 C. After 1 h, the flask is allowed to cool to room temperature. After 24 h, volatiles are removed via rotary evaporation. The resulting solid is stirred in 360 mL of boiling dioxane. The suspension is then filtered out while the solvent is still hot, and the filtrate is let at room temperature over the weekend. Evaporation of the filtrate gives a white crude product (15.7 g). The crude is recrystallized from 140 mL of boiling water to give the expected compound.
[0641] 1.2.30. Int 139
##STR00084##
[0642] Int 139 is prepared from (S)-2-Amino-pentanedioic acid using same experimental procedure as for preparation of Int 136.
TABLE-US-00002 TABLE II Illustrative intermediate for the synthesis of illustrative compounds of the invention
TABLE-US-00003 TABLE III Illustrative compounds of the invention trans:
TABLE-US-00004 TABLE IV NMR of illustrative compounds of the invention Cpd NMR 56 .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 10.62 (1H, br. s), 7.76 (1H, m), 6.56-6.46 (2H, m), 4.89-4.80 (1H, m), 3.86 (3H, s), 3.66-3.54 (4H, m), 3.42-3.34 (1H, m), 3.30-3.23 (1H, m), 3.20-3.06 (4H, m), 2.86-2.75 (1H, m), 2.07 (1H, dd), 1.65 (1H, d), 1.18 (3H, s) 68 .sup.1H NMR (500 MHz, DMSO-d.sub.6) ppm 10.63 (1H, s), 7.92 (1 H, s), 7.09 (1H, dd), 7.05 (1H, t), 6.92 (1H, dd), 3.80 (3H, s), 3.64-3.59 (2H, m), 3.59-3.53 (2H, m), 3.07-3.01 (2H, m), 3.00-2.93 (2H, m), 2.38 (1H, dt), 2.21 (1H, dt), 1.82 (2H, t), 1.27 (3H, s) 78 .sup.1H NMR (400 MHz, DMSO-d.sub.6) d ppm 10.6 (1H, s), 7.91 (1H, s), 6.94-6.81 (2H, m), 6.67 (1H, td), 3.79 (3H, s), 3.60-3.40 (4H, m), 3.00-2.75 (4H, m), 2.41-2.30 (1H, m), 2.23-2.11 (1H, m), 1.87-1.74 (2H, m), 1.26 (3H, s) 98 .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 13.0-12.0 (1H, b), 10.61 (1H, s), 7.90-7.84 (1H, m), 7.30-7.15 (1H, m), 7.97-6.66 (2H, m), 4.28-3.92 (2H, m), 3.85-3.72 (1H, m), 3.70-3.20 (2H, m), 3.20-2.80 (2H, m), 2.82-2.72 (1H, m), 2.69-2.52 (3H, m), 2.48-2.13 (2H, m), 1.93-1.73 (2H, m), 1.15 (3H, t), 0.97-0.78 (3H, m) 105 .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 10.63 (1H, s), 7.94 (1 H, m), 6.49-6.44 (2H, m), 4.28-3.99 (2H, m), 3.80 (3H, s), 3.83-3.75 (0.5H, m), 3.63-3.57 (0.5H, m), 3.49-3.42 (0.5H, m), 3.35-3.22 (1.5H, m), 3.08-2.84 (2H, m), 2.48-2.29 (1H, m), 2.25 (3H, s), 2.26-2.13 (1H, m), 1.91-1.75 (2H, m), 1.29-1.24 (3H, m), 0.94-0.81 (3H, m) 116 .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 12.4-12.1 (1H, b), 10.74 (1H, s), 7.93-7.87 (1H, s), 7.19 (1H, d), 6.88-6.81 (1H, m), 6.78-6.70 (1H, m), 4.26-4.05 (1H, m), 4.04-3.94 (1H, m), 3.83-3.21 (3H, m), 3.09-2.83 (2H, m), 2.63 (2H, q), 2.48-2.13 (3H, m), 2.13-2.02 (1H, m), 1.93-1.77 (4H, m), 1.15 (3H, t), 0.93-0.81 (3H, m) 119 .sup.1H NMR (400 MHz, CD.sub.3OD) ppm 6.46-6.36 (2H, m), 4.41-4.33 (0.5H, m), 4.30-4.21 (0.5H, m), 4.10-3.98 (1H, m), 3.96-3.88 (0.5H, m), 3.87 (3H, s), 3.78-3.69 (0.5H, m), 3.61-3.53 (0.5H, m), 3.43-3.32 (1.5, m), 3.23-3.10 (1H, m), 3.09-3.02 (1H, m), 2.60-2.24 (2H, m), 2.10-1.97 (2H, m), 1.42 (3H, d), 1.04 (1.5H, d), 0.98 (1.5H, d)
Biological Examples
Example 2. In Vitro Assays
[0643] 2.1. hADAMTS-1
[0644] The basis for the assay is the cleavage of the substrate 5(6)-Fluorescein-NH-AELQGRPISIAK-5(6)-TAMRA (SEQ ID NO: 1) by human ADAMTS1
[0645] For the dose response (10 point), 4 L of a dilution series of compound (2 mM highest concentration, 1/5 dilution in DMSO further diluted 1 in 10 in water corresponding to a final highest concentration of 20 M), is transferred to 384 well Fluotrac 200 plate (Greiner, cat#781076) and incubated at room temperature for 30 min with a 26 L buffer solution (50 mM MOPS pH7; 50 mM NaCl; 5 mM CaCl.sub.2; 0.05% CHAPS; 5 M ZnCl.sub.2) containing hADAMTS1 (0.38 ng/L, R&D SYSTEMS INC., Cat#2197-AD)) (it will be appreciated by the skilled person that the potency read out is independent of the enzyme concentration).
[0646] The reaction is initiated by adding to the assay plate 5(6)-Fluorescein-NH-AELQGRPISIAK-5(6)-TAMRA (SEQ ID NO: 1) (10 L, 7 M, Anaspec) in the same buffer.
[0647] Finally, the fluorescence is read on the Envision (Perkin Elmer) after an incubation of 120 min at 37 C. (Excitation 485 nm, Emission 535).
[0648] 2.2. hADAMTS-4
[0649] 2.2.1. Protocol 1
[0650] The basis for the assay is the cleavage of the substrate TBIS-1 (5-FAM-TEGEARGSVILLK (5TAMRA)K-NH.sub.2) (SEQ ID NO: 2) by human ADAMTS4.
[0651] For the dose response (10 point), 4 L of a dilution series of compound (2 mM highest concentration, 1/5 dilution in DMSO further diluted 1 in 10 in water corresponding to a final highest concentration of 20 M), is transferred to 384 well Fluotrac 200 plate (Greiner, cat#781076) and incubated at room temperature for 30 min with a 26 L buffer solution (50 mM Hepes pH7.5, 100 mM NaCl, 5 mM CaCl.sub.2, 0.1% CHAPS, 5% glycerol) containing hADAMTS4 (0.325 ng/L) (it will be appreciated by the skilled person that the potency read out is independent of the enzyme concentration).
[0652] The reaction is initiated by adding to the assay plate TBIS-1 (10 L, 4.5 M, Anaspec) in the same buffer.
[0653] Finally, the fluorescence is read on the Envision (Perkin Elmer) after an incubation of 60 min at room temperature (Excitation 485 nm, emission 535).
[0654] The IC.sub.50 measured for illustrative compounds of the invention is reported in the table below.
TABLE-US-00005 TABLE V ADAMTS4 potency of illustrative compounds of the invention Cpd # IC.sub.50 (nM) 1 * 2 *** **** 0.001-25 nM *** >25-50 nM ** >50-100 nM * >100 nM
[0655] 2.2.2. Protocol 2
[0656] The basis for the assay is the cleavage of the substrate TBIS-1 (5 FAM-TEGEARGSVILLK (5TAMRA)K-NH.sub.2) (SEQ ID NO: 2) by human ADAMTS4.
[0657] For the dose response (10 point), 4 L of a dilution series of compound (2 mM highest concentration, 1/5 dilution in DMSO further diluted 1 in 10 in water corresponding to a final highest concentration of 20 M), is transferred to 384 well Fluotrac 200 plate (Greiner, cat#781076) and incubated at room temperature for 30 min with a 26 L buffer solution (50 mM Hepes pH 7.5, 100 mM NaCl, 5 mM CaCl.sub.2, 0.1% CHAPS) containing hADAMTS4 (0.38 ng/L) (it will be appreciated by the skilled person that the potency read out is independent of the enzyme concentration).
[0658] The reaction is initiated by adding to the assay plate TBIS-1 (10 L, 4.5 M, Anaspec) in the same buffer.
[0659] Finally, the fluorescence is read on the Envision (Perkin Elmer) after an incubation of 180 min at 37 C. (Excitation 485 nm, emission 535).
[0660] The IC.sub.50 measured for illustrative compounds of the invention is reported in the table below.
TABLE-US-00006 TABLE VI ADAMTS4 potency of illustrative compounds of the invention Cpd # IC.sub.50 (nM) 3 * 8 ** 10 * 11 * 14 * 15 ** 17 * 18 * 19 * 20 * 21 ** 22 * 23 ** 24 * 25 * 26 **** 27 *** 28 * 29 *** 30 * 31 * 34 * 36 * 37 *** 38 * 39 * 40 *** 41 * 42 *** 43 *** 46 *** 47 *** 48 * 49 ** 50 * 51 * 52 * 53 * 54 *** 55 * 56 *** 57 ** 58 * 59 *** 60 * 61 * 62 ** 63 * 64 * 65 ** 71 * 74 * 75 * 79 * 80 * 85 * 86 * 89 * 90 *** 93 * 96 * 99 * 105 ** 106 * 107 * 109 * 112 ** 114 * 115 * 116 * 117 * 118 * 119 * 120 ** 121 ** 122 ** 123 ** 124 * 125 ** 128 * 129 * 132 * 133 * 135 * 138 * 139 * 142 * 143 * 144 * 145 * 146 * 147 * 148 * 150 * **** 0.001-25 nM *** >25-50 nM ** >50-100 nM * >100 nM
[0661] 2.3. Rat ADAMTS-5
[0662] The basis for the assay is the cleavage of the substrate TBIS-1 (5 FAM-TEGEARGSVILLK (5TAMRA)K-NH.sub.2) (SEQ ID NO: 2) by rnADAMTS-5 (1-564-6H).
[0663] For the dose response (10 point), 4 L of a dilution series of compound (2 mM highest concentration, 1/5 dilution in DMSO further diluted 1 in 10 in water, corresponding to a final highest concentration of 20 M), is transferred to 384 well Fluotrac 200 plate (Greiner, cat#781076) and incubated at room temperature for 30 min with a 26 L buffer solution (50 mM TRIS pH7.5, 100 mM NaCl, 5 mM CaCl.sub.2, 0.1% CHAPS) containing rnADAMTS-5 (0.5 ng/L) (it will be appreciated by the skilled person that the potency read out is independent of the enzyme concentration).
[0664] The reaction is initiated by adding to the assay plate TBIS-1 (10 L, 4.5 M, Anaspec) in the same buffer.
[0665] Finally, the fluorescence is read on the Envision (Perkin Elmer) after an incubation of 120 min at 37 C. (Excitation 485 nm, emission 535).
[0666] The IC.sub.50 measured for illustrative compounds of the invention is reported in the table below.
TABLE-US-00007 TABLE VII Rat ADAMTS-5 potency of illustrative compounds of the invention Cpd # IC.sub.50 (nM) 2 * 6 * 8 * 9 * 10 * 11 * 12 * 13 * 14 * 15 ** 17 * 18 * 21 **** 23 ** 24 ** 25 * 26 **** 27 *** 28 ** 29 *** 30 * 31 * 34 * 36 * 37 **** 38 ** 39 ** 40 * 41 **** 42 **** 43 **** 52 ** 53 * 54 **** 56 *** 59 **** 85 * 86 * 129 * 130 * 131 * 132 * 133 * 135 * 138 * 139 ** 142 * 144 ** **** 0.001-25 nM *** >25-50 nM ** >50-100 nM * >100 nM
[0667] 2.4. hADAMTS-5
[0668] 2.4.1. Protocol 1
[0669] The basis for the assay is the cleavage of the substrate TBIS-1 (5 FAM-TEGEARGSVILLK (5TAMRA)K-NH.sub.2) (SEQ ID NO: 2) by human ADAMTS-5.
[0670] For the dose response (10 point), 4 L of a dilution series of compound (2 mM highest concentration, 1/5 dilution in DMSO further diluted 1 in 10 in water, corresponding to a final highest concentration of 20 M), is transferred to 384 well Fluotrac 200 plate (Greiner, cat#781076) and incubated at room temperature for 30 min with a 26 L buffer solution (50 mM Hepes pH7.5, 100 mM NaCl, 5 mM CaCl.sub.2, 0.1% CHAPS, 5% glycerol) containing hADAMTS-5 (0.5 ng/L) (it will be appreciated by the skilled person that the potency read out is independent of the enzyme concentration).
[0671] The reaction is initiated by adding to the assay plate TBIS-1 (10 L, 4.5 M, Anaspec) in the same buffer.
[0672] Finally, the fluorescence is read on the Envision (Perkin Elmer) after an incubation of 60 min at Room Temperature (Excitation 485 nm, emission 530).
[0673] 2.4.2. Protocol 2
[0674] The basis for the assay is the cleavage of the substrate TBIS-1 (5 FAM-TEGEARGSVILLK (5TAMRA)K-NH.sub.2) (SEQ ID NO: 2) by human ADAMTS-5.
[0675] For the dose response (10 point), 4 L of a dilution series of compound (2 mM highest concentration, 1/5 dilution in DMSO further diluted 1 in 10 in water, corresponding to a final highest concentration of 20 M), is transferred to 384 well Fluotrac 200 plate (Greiner, cat#781076) and incubated at room temperature for 30 min with a 26 L buffer solution (50 mM Hepes pH7.5, 100 mM NaCl, 5 mM CaCl.sub.2, 0.1% CHAPS 1) containing hADAMTS-5 (1 ng/L, affinity purified, followed by overnight digestion of 6His tag by thrombin and dialysis) (it will be appreciated by the skilled person that the potency read out is independent of the enzyme concentration).
[0676] The reaction is initiated by adding to the assay plate TBIS-1 (10 L, 4.5 M, Anaspec) in the same buffer.
[0677] Finally, the fluorescence is read on the Envision (Perkin Elmer) after an incubation of 45 min at 37 C. (Excitation 485 nm, emission 530).
[0678] The IC.sub.50 measured for illustrative compounds of the invention is reported in the table below.
[0679] hADAMTS-5 potency of illustrative compounds of the invention
[0680] The IC.sub.50 measured for illustrative compounds of the invention is reported in the table below.
TABLE-US-00008 TABLE VIII ADAMTS5 potency of illustrative compounds of the invention Cpd # IC.sub.50 (nM) 1 * 2 * 3 * 4 * 5 * 6 * 7 * 8 * 9 * 10 * 11 * 12 * 13 * 14 * 15 ** 16 * 17 * 18 ** **** 0.001-25 nM *** >25-50 nM ** >50-100 nM * >100 nM
[0681] 2.4.3. Protocol 3
[0682] The basis for the assay is the cleavage of the substrate TBIS-1 (5 FAM-TEGEARGSVILLK (5TAMRA)K-NH.sub.2) (SEQ ID NO: 2) by human ADAMTS-5.
[0683] For the dose response (10 point), 4 L of a dilution series of compound (2 mM highest concentration, 1/5 dilution in DMSO further diluted 1 in 10 in water, corresponding to a final highest concentration of 20 M), is transferred to 384 well Fluotrac 200 plate (Greiner, cat#781076) and incubated at room temperature for 30 min with a 26 L buffer solution (50 mM Hepes pH 7.5, 100 mM NaCl, 5 mM CaCl.sub.2, 0.1% CHAPS) containing hADAMTS-5 (0.63 ng/L) (it will be appreciated by the skilled person that the potency read out is independent of the enzyme concentration).
[0684] The reaction is initiated by adding to the assay plate TBIS-1 (10 L, 4.5 M, Anaspec) in the same buffer.
[0685] Finally, the fluorescence is read on the Envision (Perkin Elmer) after an incubation of 90 min at 37 C. (Excitation 485 nm, emission 530).
[0686] The IC.sub.50 measured for illustrative compounds of the invention is reported in the table below.
TABLE-US-00009 TABLE IX hADAMTS-5 potency of illustrative compounds of the invention Cpd # IC.sub.50 (nM) 4 * 7 * 9 * 10 * 14 * 15 ** 17 * 18 ** 19 * 20 * 21 **** 22 * 23 ** 24 *** 25 * 26 **** 27 *** 28 * 29 *** 30 * 31 * 32 * 33 * 34 * 35 * 36 * 37 **** 38 * 39 ** 40 ** 41 *** 42 **** 43 **** 44 ** 45 * 46 *** 47 *** 48 ** 49 *** 50 * 51 * 52 ** 53 ** 54 **** 55 ** 56 *** 57 *** 58 * 59 **** 60 * 61 *** 62 **** 63 *** 64 ** 65 **** 66 * 69 * 70 * 71 * 73 * 74 * 75 ** 76 * 78 * 79 * 80 * 85 * 86 * 89 * 90 **** 91 * 93 * 94 * 95 * 96 * 98 * 99 * 105 * 106 * 107 * 109 * 110 * 111 * 112 * 113 * 114 * 115 * 116 * 117 ** 118 * 119 **** 120 **** 121 *** 122 **** 123 **** 124 *** 125 *** 126 *** 127 * 128 * 129 * 130 * 131 * 132 * 133 * 134 * 135 * 136 * 137 * 138 * 139 * 140 * 142 * 143 * 144 ** 145 ** 146 * 147 * 148 *** 149 * 150 * 151 * **** 0.001-25 nM *** >25-50 nM ** >50-100 nM * >100 nM
[0687] 2.5. hADAMTS-6
[0688] 2.5.1. Preparation of hADAM-TS6 Protein
[0689] 2.5.1.1. ADAMTS-6 Protein Expression.
[0690] The coding sequence from amino acid 1 to 843 of the human ADAMTS6 protein (UniProtKB/Swiss-Prot Q9UKP5, SEQ ID NO: 4) is cloned into the pYD7 vector (vectors and the expression system are described in (Durocher et al., 2002; Loignon et al., 2008; Shi et al., 2005)) (CNRC-NRC) in C-terminal fusion with a sequence containing Tev-GGGGGS-Thr-6his (SEQ ID NO: 5) (pYD7-HsADAM-TS6(1-843)-Tev-GGGGGS-Thr-6his construct). Durocher's 293-6E cells (CNRC-NRC) are grown in suspension at 37 C. in a humidified incubator containing 5% CO.sub.2 at 37 C. in F17 culture medium (Invitrogen # A1383501) supplemented with 10 mL/L of 10% Pluronic F68 solution (0.1% final) (Invitrogen #24040032), 20 mL/L of 200 mM L-Glutamine solution (4 mM final) (Lonza # BE17-605E) and 25 g/mL G418 (PAA laboratories # P11-012).
[0691] Transfection is performed according to the protocol given by CNRC-NRC for the 293-6E cells using PEI (Polysciences #23966-2). Cells are diluted to 3.105 cells/mL three days before transfection. On the day of transfection, the cell density is adjusted to 17.105 cells/mL. A DNA/PEI mix is prepared according to CNRC-NRC recommendations with a ratio of DNA/PEI of 1 for 2, meaning, 300 g of pYD7-HsADAM-TS6(1-843)-Tev-GGGGGS-Thr-6his construct and 600 g PEI for 300 mL of final transfection volume. The day after transfection, Heparin (Sigma # H3149) and TN1 (Teckniscience Inc. #19553) is added to get 0.1 g/L and 0.5% final concentrations respectively.
[0692] The conditioned medium is harvested 168 h post-transfection by centrifugation for 20 min at 5000g at 4 C. The expression level of secreted HsADAM-TS6 is estimated at 1 mg/L in the conditioned medium (CM) by Western blotting analysis using anti His tag primary antibody. Conditioned medium is either used from fresh or frozen production at 20 C. for later use.
[0693] 2.5.1.2. Purification of HsADAM-TS6 Protein
[0694] 10 buffer A (500 mM Tris pH 8, 4 M NaCl, 10 mM CaCl.sub.2, 10 M ZnCl.sub.2, 0.5% CHAPS, 200 mM imidazole) is slowly added to the conditioned medium under magnetic stirring, giving Buffered Conditioned Medium (BCM). This sample is clarified by centrifugation at 15,000g for 60 min at 4 C. in a Beckman 16.250 rotor, providing the starting sample.
[0695] Affinity purification is performed using AKTA protein purification system (GE Healthcare) in 4 C. cabinet. 400 mL of the start sample is loaded onto each HisTrap HP 5 mL column (GE Healthcare #17-5248-02) at a flow rate of 5 mL/min. The resin is then washed with equilibration buffer B (50 mM Tris (pH 8), 0.5 M NaCl, 1 mM CaCl.sub.2, 1 M ZnCl.sub.2, 20 mM imidazole, 0.05% CHAPS) until the absorbance at 280 nm returns to zero. The elution is then performed with 70 mM imidazole (elution of contaminant proteins) and subsequently 300 mM imidazole (elution of the target protein). The fractions containing the non-mature (1-843) ADAMTS6 protein are pooled and subjected to 2 dialyses against 100 volumes of dialysis buffer C (50 mM Tris pH 8+100 mM NaCl+1 mM CaCl.sub.2+1 M ZnCl.sub.2+0.1% Polyoxyethyleneglycol Dodecyl Ether [Brij 35]) using 50 kDa cut off membrane (SpectraPor #132 542). This allows to decrease the NaCl concentration for efficient furin cleavage.
[0696] 2.5.1.3. Maturation of HsADAM-TS6 Protein
[0697] In vitro maturation of human ADAMTS6 protein is performed using furin (R&D #1503-SE) (3 ng furin/g of target protein), in buffer D (25 mM Tris pH 9+100 mM NaCl+1 mM CaCl.sub.2+1 M ZnCl.sub.2+0.5% Brij 35). Incubation is conducted overnight at room temperature.
[0698] The furin cleaved sample is subsequently dialyzed for 1 hour against 100 volumes of dialysis buffer C. Finally, 10% glycerol is added to the final batch which is subsequently ultracentrifuged for 30 minutes at 35,000 rpm using a Beckman SW41Ti rotor and stored at 80 C.
[0699] Following this procedure the final yield reached an average of 0.05 to 0.1 mg matured human ADAMTS6 (SEQ ID NO: 6)/L of culture medium.
[0700] 2.5.2. hADAMTS-6 Assay
[0701] The IC.sub.50 value for test compounds can be determined in a fluorescent based protease assay.
[0702] The basis for the assay is the cleavage of the substrate 520 MMP FRET substrate XII by human ADAMTS6.
[0703] The substrate contains sequence Arg-Pro-Lys-Pro-Tyr-Ala-Nva-Trp-Met-Lys (SEQ ID NO: 7) with attached a fluorescent probe and a quencher. Without ADAMTS6, the emission of the probe is quenched. If the substrate is cleaved by ADAMTS6, the emission of the probe is not quenched anymore. Inhibition of ADAMTS6 activity, will result in a decrease of the signal.
[0704] To perform the assay, 4 L of a dilution series of compound in water, starting from 20 M highest concentration, 1/5 dilution, is added to the wells. 2 ng ADAMTS6 enzyme is diluted in 25 mM Tris pH8.0, 0.05% CHAPS, 2.5 mM CaCl.sub.2 in a total volume of 26 L (final concentration 0.73 nM). The reaction is started by addition of 10 L of 1 M 520 MMP FRET Substrate XII (final concentration, diluted in same buffer as described above) and the mixture is incubated at 37 C. for 2 h. The negative control (0% inhibition) is 1% DMSO and the positive control (100% inhibition) is 10 M Prinomastat in 1% DMSO. After this incubation, cleavage of the substrate is measured using the Envision (Perkin Elmer, exc485/em530).
[0705] The IC.sub.50 measured for illustrative compounds of the invention is reported in the table below.
TABLE-US-00010 TABLE X hADAMTS-6 potency of illustrative compounds of the invention Cpd # IC.sub.50 (nM) 1 * 2 * 3 * 4 * 5 * 6 * 7 * 8 * 9 ** 10 *** 11 * 12 * 13 *** 14 *** 15 * 16 * 17 * 18 **** 19 ** 20 **** 21 **** 22 **** 23 *** 24 **** 25 *** 26 **** 27 **** 28 *** 29 **** 30 **** 31 **** 32 * 33 * 34 *** 35 * 36 *** 37 **** 38 **** 39 **** 40 **** 41 **** 42 **** 43 **** 44 **** 45 * 46 **** 47 **** 48 **** 49 **** 50 **** 51 **** 52 **** 53 **** 54 **** 55 **** 56 **** 57 **** 58 *** 59 **** 60 * 61 **** 62 **** 63 **** 64 ** 65 **** 66 ** 67 ** 68 * 69 * 70 * 71 *** 72 * 73 * 74 **** 75 **** 76 * 77 * 78 * 79 **** 80 **** 81 * 82 * 83 * 84 * 85 **** 86 **** 87 ** 88 *** 89 **** 90 **** 91 * 92 **** 93 **** 94 **** 95 **** 96 **** 97 *** 98 **** 99 **** 100 **** 101 **** 102 * 103 * 104 **** 105 **** 106 **** 107 * 108 * 109 * 110 **** 111 **** 112 **** 113 * 114 **** 115 **** 116 **** 117 **** 118 **** 119 **** 120 **** 121 **** 122 **** 123 **** 124 **** 125 **** 126 **** 127 * 128 ** 129 **** 130 * 131 * 132 *** 133 **** 134 ** 135 **** 136 * 137 * 138 * 139 **** 140 * 142 ** 143 **** 144 **** 145 **** 146 * 147 **** 148 **** 149 ** 150 **** 151 *** **** 0.001-25 nM *** >25-50 nM ** >50-100 nM * >100 nM
[0706] 2.6. hTACE
[0707] The basis for the assay is the cleavage of the substrate 5FAM-LAQAVRSSSRK-5TAMRA (SEQ ID NO: 3) (Anaspec, custom 34891) by human TACE (R&D SYSTEMS INC., Cat#930-ADB).
[0708] For the dose response (10 point), 4 L of a dilution series of compound (2 mM highest concentration, 1/5 dilution in DMSO further diluted 1 in 10 in water, corresponding to a final highest concentration of 20 M), is transferred to 384 well Fluotrac 200 plate (Greiner, cat#781076) and incubated at room temperature for 30 min with a 26 L buffer solution (25 mM Tris pH8.0, 2.5 M ZnCl.sub.2, 0.01% CHAPS) containing TACE (0.05 ng/L) (it will be appreciated by the skilled person that the potency read out is independent of the enzyme concentration).
[0709] The reaction is initiated by adding to the assay plate 5FAM-LAQAVRSSSRK-5TAMRA (5 L, 5 M, Anaspec) in the same buffer.
[0710] Finally, the fluorescence is read on the Envision (Perkin Elmer) after an incubation of 75 min at room temperature (Excitation 485 nm, Emission 530).
[0711] The IC.sub.50 measured for illustrative compounds of the invention is reported in the table below.
TABLE-US-00011 TABLE XI hTACE potency of illustrative compounds of the invention Cpd # IC.sub.50 (nM) 1 * 2 * 3 * 6 * 8 * 9 * 10 * 11 * 12 * 13 * 14 * 15 * 17 * 18 * 19 * 20 * 21 * 22 * 23 * 24 * 25 * 26 * 27 * 28 * 29 * 30 * 31 * 34 * 36 * 37 * 38 * 39 * 40 * 41 * 42 * 43 * 46 * 47 * 48 * 49 * 50 * 51 * 52 * 53 * 54 * 55 * 56 * 57 * 58 * 59 * 60 * 61 * 62 * 63 * 64 * 65 * 66 * 69 * 70 * 71 * 73 * 74 * 75 * 76 * 78 * 79 * 80 * 85 * 86 * 89 * 90 * 91 * 93 * 94 * 95 * 96 * 99 * 105 * 106 * 107 * 109 * 112 * 114 * 115 * 116 * 117 * 118 * 119 * 120 * 121 * 122 * 123 * 124 * 125 * 128 * 129 * 132 * 133 * 135 * 138 * 139 * 142 * 143 * 144 * 145 * 146 * 147 * 148 * 149 * 150 * 151 * **** 0.001-25 nM *** >25-50 nM ** >50-100 nM * >100 nM
[0712] 2.7. hMMP1
[0713] Inhibition of the proteases human MMP1 was determined at REACTION BIOLOGY (Reaction Biology Corp. 1 Great Valley Parkway, Suite 2 Malvern, Pa. 19355, USA) in fluorescent based biochemical assays. The protease activities were monitored as a time-course measurement of the increase in fluorescence signal from fluorescently-labeled peptide substrates, and initial linear portion of slope (signal/min) was analyzed.
[0714] To determine the IC.sub.50, a compound is tested starting from 100 nM (highest dilution) with a 1/3 dilution.
[0715] 2.8. hMMP2
[0716] 2.8.1. Protocol 1
[0717] The basis for the assay is the cleavage of the substrate 520 MMP fret substrate XV (Anaspec, Catalog #: AS-60582-01) by human MMP2 (R&D SYSTEMS INC. Systems Inc., Cat#902-MP).
[0718] For the dose response (10 point), 4 L of a dilution series of compound (2 mM highest concentration, 1/5 dilution in DMSO further diluted 1 in 10 in water, corresponding to a final highest concentration of 20 M), is transferred to 384 well Fluotrac 200 plate (Greiner, cat#781076) and incubated at room temperature for 30 min with a 26 L buffer solution (50 mM Tris pH 7.5, 10 mM, CaCl.sub.2, 150 mM NaCl, 0.05% Brij35) containing preactivated MMP2 (0.0125 ng/L) (it will be appreciated by the skilled person that the potency read out is independent of the enzyme concentration). Human MMP2 is preactivated by incubated the enzyme in the same buffer complemented with 1 mM freshly prepared p-Aminophenylmercuric acetate (AMPA) for 1 h at 37 C.
[0719] The reaction is initiated by adding to the assay plate 520 MMP fret substrate XV (10 L, 4 M, Anaspec) in the same buffer.
[0720] Finally, the fluorescence is read on the Envision (Perkin Elmer) after an incubation of 30 min at room temperature (Excitation 485 nm, Emission 530).
[0721] 2.8.2. Protocol 2
[0722] The basis for the assay is the cleavage of the substrate 390 MMP FRET substrate I (Anaspec, Catalog n#: AS-27076) by human MMP2 (R&D SYSTEMS INC., Cat#902-MP).
[0723] For the dose response (10 point), 4 L of a dilution series of compound (2 mM highest concentration, 1/5 dilution in DMSO further diluted 1 in 10 in water, corresponding to a final highest concentration of 20 M), is transferred to 384 well Fluotrac 200 plate (Greiner, cat#781076) and incubated at room temperature for 30 min with a 26 L buffer solution (45 mM Tris pH 7.5, 9 mM CaCl.sub.2, 135 mM NaCl, 0.045% Brij35) containing MMP2 (0.03 ng/L) (it will be appreciated by the skilled person that the potency read out is independent of the enzyme concentration).
[0724] The reaction is initiated by adding to the assay plate 390 MMP FRET substrate I (10 L, 2.5 M, Anaspec) in the same buffer.
[0725] Finally, the fluorescence is read on the Envision (Perkin Elmer) after an incubation of 30 min at room temperature (Excitation 485 nm, Emission 530).
[0726] 2.9. hMMP8
[0727] Inhibition of the human MMP8 protease is determined at REACTION BIOLOGY (Reaction Biology Corp. 1 Great Valley Parkway, Suite 2 Malvern, Pa. 19355, USA; cat# MMP8) in fluorescence based biochemical assays. The protease activity is monitored as a time-course measurement of the increase in fluorescence signal from fluorescently-labeled peptide substrates, and the slope (signal/min) of the initial linear portion is measured.
[0728] The basis for the assay is the cleavage of the substrate 520 MMP FRET Substrate XIV (Anaspec, cat# AS-60581) by human MMP8 (Enzo, cat# SE-255) in a buffer solution (50 mM HEPES pH 7.5, 10 mM CaCl.sub.2, 0.01% Brij-35, 0.1 mg/mL BSA).
[0729] A 100% DMSO dilution series of test compound (10 final concentrations starting from 30 M highest concentration, with 1/3 serial dilutions) is added to MMP8 in buffer solution and incubated at room temperature for 5-15 min (it will be appreciated by the skilled person that the potency read out is independent of the enzyme concentration). The reaction is then initiated by adding 520 MMP FRET Substrate XIV (5 M final concentration) in the same buffer.
[0730] Fluorescence is read at 5 min intervals for 2 h with an Envision (Perkin Elmer) at room temperature (Excitation 485 nm, Emission 520 nm). The slope of the initial linear portion of the fluorescence signal curve is then calculated by using Excel. Percent protease activity is calculated relative to a no inhibitor DMSO control defined as 100% activity. IC.sub.50 curve fits are performed using Prism software.
[0731] 2.10. hMMP12
[0732] Inhibition of the human MMP12 protease is determined at REACTION BIOLOGY (Reaction Biology Corp. 1 Great Valley Parkway, Suite 2 Malvern, Pa. 19355, USA; cat# MMP12) in fluorescence based biochemical assays. The protease activity is monitored as a time-course measurement of the increase in fluorescence signal from fluorescently-labeled peptide substrates, and the slope (signal/min) of the initial linear portion is measured.
[0733] The basis for the assay is the cleavage of the substrate 520 MMP FRET Substrate XIV (Anaspec, cat# AS 60581) by human MMP12 (Enzo, cat# SE-138) in a buffer solution (50 mM HEPES pH 7.5, 10 mM CaCl.sub.2, 0.01% Brij-35, 0.1 mg/mL BSA).
[0734] A 100% DMSO dilution series of test compound (10 final concentrations starting from 30 M highest concentration, with 1/3 serial dilutions) is added to MMP12 in buffer solution and incubated at room temperature for 5-15 min (it will be appreciated by the skilled person that the potency read out is independent of the enzyme concentration). The reaction is then initiated by adding 520 MMP FRET Substrate XIV (5 M final concentration) in the same buffer.
[0735] Fluorescence is read at 5 min intervals for 2 h with an Envision (Perkin Elmer) at room temperature (Excitation 485 nm, Emission 520 nm). The slope of the initial linear portion of the fluorescence signal curve is then calculated by using Excel. Percent protease activity is calculated relative to a no inhibitor DMSO control defined as 100% activity. IC.sub.50 curve fits are performed using Prism software.
[0736] 2.11. hMMP13
[0737] 2.11.1. Protocol 1
[0738] The basis for the assay is the cleavage of the substrate 390 MMP FRET Substrate I (Anaspec Cat# AS-27076) by human MMP13 (Chemicon, Cat#CC068).
[0739] For the dose response (10 point), 4 L of a dilution series of compound (20 M highest concentration, 1/5 dilution in water), is transferred to 384 well Fluotrac 200 plate (Greiner, cat#781076) and incubated at room temperature for 30 min with a 26 L buffer solution (50 mM Tris pH7.5, 150 mM NaCl, 10 mM CaCl.sub.2, 0.05% CHAPS, 5 M ZnCl.sub.2) containing MMP13 (0.01 ng/L) (it will be appreciated by the skilled person that the potency read out is independent of the enzyme concentration). Human MMP13 is preactivated by incubated the enzyme in the same buffer complemented with 1 mM freshly prepared p-Aminophenylmercuric acetate (AMPA) for 1 h at 37 C.
[0740] The reaction is initiated by adding to the assay plate 390 MMP FRET Substrate I (10 L, 2.5 M) in the same buffer.
[0741] Finally, the fluorescence is read on the Envision (Perkin Elmer) after an incubation of 45 min at room temperature (Excitation 485 nm, Emission 530).
[0742] The IC.sub.50 measured for illustrative compounds of the invention is reported in the table below.
TABLE-US-00012 TABLE XII hMMP-13 potency of illustrative compounds of the invention Cpd # IC.sub.50 (nM) 3 * 6 * 8 * 9 * 10 * 11 * 12 * 13 * 14 * 15 * 17 * 18 * 19 * 20 * 21 * 22 * 23 * 24 * 25 * 26 * 27 * 28 * 29 * 30 * 31 * 34 * 36 * 37 * 38 * 39 * 40 * 41 * 42 * 43 * 46 * 47 * 48 * 50 * 53 * 128 * 129 * 132 * 133 * 135 * 138 * 139 * 142 * 143 * 144 * 145 * **** 0.001-25 nM *** >25-50 nM ** >50-100 nM * >100 nM
[0743] 2.11.2. Protocol 2
[0744] The basis for the assay is the cleavage of the substrate 390 MMP FRET Substrate I (Anaspec Cat# AS-27076) by human MMP13 (Enzo Life Sciences, Cat#BML-SE493).
[0745] For the dose response (10 point), 4 L of a dilution series of compound (20 M highest concentration, 1/5 dilution in water), is transferred to 384 well Fluotrac 200 plate (Greiner, cat#781076) and incubated at room temperature for 30 min with a 26 L buffer solution (50 mM Tris pH7.5, 150 mM NaCl, 10 mM CaCl.sub.2, 0.05% CHAPS, 5 M ZnCl.sub.2) containing MMP13 (0.01 ng/L) (it will be appreciated by the skilled person that the potency read out is independent of the enzyme concentration). Human MMP13 is preactivated by incubated the enzyme in the same buffer complemented with 1 mM freshly prepared p-Aminophenylmercuric acetate (AMPA) for 1 h at 37 C.
[0746] The reaction is initiated by adding to the assay plate 390 MMP FRET Substrate I (10 L, 2.5 M) in the same buffer.
[0747] Finally, the fluorescence is read on the Envision (Perkin Elmer) after an incubation of 45 min at room temperature (Excitation 485 nm, Emission 530).
[0748] The IC.sub.50 measured for illustrative compounds of the invention is reported in the table below.
TABLE-US-00013 TABLE XIII hMMP-13 potency of illustrative compounds of the invention Cpd # IC.sub.50 (nM) 18 * 51 * 52 * 53 * 54 * 55 * 56 * 57 * 58 * 59 * 61 * 62 * 63 * 64 * 65 * 71 * 74 * 75 * 79 * 80 * 85 * 86 * 89 * 90 * 93 * 96 * 99 * 105 * 106 * 107 * 109 * 112 * 114 * 115 * 116 * 117 * 118 * 119 * 120 * 121 * 122 * 123 * 124 * 125 * 146 * 147 * 148 * 150 * **** 0.001-25 nM *** >25-50 nM ** >50-100 nM * >100 nM
[0749] 2.11.3. Protocol 3
[0750] The basis for the assay is the cleavage of the substrate 520 MMP-fret substrate XV (Anaspec, Catalog #: AS-60582-01) by human MMP13 (Chemicon, Cat# CC068).
[0751] For the dose response (10 point), 4 L of a dilution series of compound (2 mM highest concentration, 1/5 dilution in DMSO further diluted 1 in 10 in water, corresponding to a final highest concentration of 20 M), is transferred to 384 well Fluotrac 200 plate (Greiner, cat#781076) and incubated at room temperature for 30 min with a 26 L buffer solution (50 mM Tris pH7.5, 150 mM NaCl, 10 mM CaCl.sub.2, 0.05% CHAPS, 5 M ZnCl.sub.2) containing MMP13 (6.25 10.sup.6 g/L) (it will be appreciated by the skilled person that the potency read out is independent of the enzyme concentration).
[0752] The reaction is initiated by adding to the assay plate 520 MMP-fret substrate XV (10 L, 4 M) in the same buffer.
[0753] Finally, the fluorescence is read on the Envision (Perkin Elmer) after an incubation of 60 min at room temperature (Excitation 485 nm, Emission 530).
[0754] 2.12. hMMP14
[0755] The basis for the assay is the cleavage of the substrate 390 MMP FRET Substrate I (Anaspec Cat# AS-27076) by human MMP14 (Biomol, Cat#SE-259).
[0756] For the dose response (10 point), 4 L of a dilution series of compound 2 mM highest concentration, 1/5 dilution in DMSO further diluted 1 in 10 in water, corresponding to a final highest concentration of 20 M), is transferred to 384 well Fluotrac 200 plate (Greiner, cat#781076) and incubated at room temperature for 30 min with a 26 L buffer solution (50 mM MOPS pH7, 5 mM CaCl.sub.2, 1 M ZnCl.sub.2, 0.1% Brij-35) containing MMP14 (0.05 ng/L) (it will be appreciated by the skilled person that the potency read out is independent of the enzyme concentration).
[0757] The reaction is initiated by adding to the assay plate 390 MMP FRET Substrate I (10 L, 2.5 M) in the same buffer.
[0758] Finally, the fluorescence is read on the Envision (Perkin Elmer) after an incubation of 60 min at room temperature (Excitation 485 nm, Emission 530).
[0759] The IC.sub.50 measured for illustrative compounds of the invention is reported in the table below.
TABLE-US-00014 TABLE XIV hMMP-14 potency of illustrative compounds of the invention Cpd # IC.sub.50 (nM) 18 * 24 * 37 * 41 * 42 * 43 * 47 * 48 * 51 * 52 * 53 * 54 * 55 * 56 * 57 * 59 * 61 * 62 * 63 * 64 * 65 * 71 * 74 * 75 * 79 * 80 * 85 * 86 * 89 * 90 * 93 * 96 * 99 * 105 * 106 * 107 * 109 * 112 * 114 * 115 * 116 * 117 * 118 * 119 * 120 * 121 * 122 * 123 * 124 * 125 * 145 * 146 * 147 * 148 * 150 * **** 0.001-25 nM *** >25-50 nM ** >50-100 nM * >100 nM
Example 3. Cellular Assays
[0760] 3.1. Mouse Explant Assay
[0761] In this assay, quantitation of glycosaminoglycans (GAGs) in the form of aggrecan fragments released from cartilage in culture is used to determine the efficacy of a test compound in preventing cartilage catabolism.
[0762] The protocol of mouse cartilage explants is described by Stanton (Stanton et al., 2011). After euthanasia, the femoral head cartilage from the right and left leg of a 3-days-old C57B16 male mouse (Janvier, 7-10 g), were placed in a 48-wells culture plate. Cell culture medium (400 L) containing human IL1 (1 ng/mL) and test compound (3 M) were added to the femoral head cartilage.
[0763] After 3 days of incubation, the supernatant is harvested and stored at 20 C. until analysis and the cartilages are digested with a papan solution at 60 C. for 24 h. Using the standard curve performed with a dose range of chondroitin sulfate, the concentration of GAG is determined in the supernatant and on the lysate using dimethylmethylene blue solution (reading at a wavelength of 590 nm).
[0764] The percentage of GAG release is calculated as follows:
The test compound effect is expressed as percent of inhibition (PIN) using the following formula:
[0765] 3.2. Human Explant Assay
[0766] In this assay, compounds are tested in human articular cartilage explants in order to evaluate their activity on aggrecan degradation induced by IL1. AGNx1 is the epitope for aggrecanase-mediated aggrecan degradation; on the other hand, AGNx2 is the epitope for MMP-mediated aggrecan degradation. Therefore quantification of AGNx1 and AGNx2 may be used to evaluate the activity of a test compound.
[0767] These studies were conducted in Nordic Bioscience (Herlev Hovedgade 207, DK-2730 Herlev, Denmark).
[0768] Human articular cartilage explants are collected from 3 nearby hospitals under an existing ethical committee application.
[0769] Full-depth cartilage explants from OA cartilage from different patients are cultured for 21 days in culture medium (DMEM/F12 with 0.5% FCS, 1% PS) containing various (positive control, untreated, and test compound at 0.1, 1 and 10 M).
[0770] The explants from each patient are cultured in a separate 96-well culture plate with 200 L/well PBS, and the 6 replicates of each treatment are distributed in a diagonal pattern on the plate. At each experimental time point (5, 12 and 19 days), supernatants are harvested from the explants cultures, and new treatment-mediums are added. The supernatants are stored at 20 C. for later biomarker analysis. The human IL13 (Sigma-Aldrich SRP3083) is used at a concentration of 10 ng/mL.
[0771] 3.3. Results
[0772] The AGNx1 and AGNx2 concentrations were determined against a standard curve. Mean and SEM were graphed using the excel software. One-way ANOVA plus Dunnett's multiple comparisons post-hoc test are used for the statistical analysis (Prism 3.03 software).
Example 4. In Vivo Assays
[0773] 4.1. In Vivo Menisectomized (MNX) Rat Model
[0774] 4.1.1. In Vivo Efficacy in the Rat MNX Model
[0775] In vivo efficacy was studied in a female Lewis meniscectomised rat (MNX) model. The MNX rat model is a well-validated disease model of osteoarthritis (Bendele, 2001; Janusz et al., 2002; Pritzker et al., 2006).
[0776] 4.1.2. Experimental Procedures
[0777] 4.1.2.1. Surgery and Dosing
[0778] Osteoarthritis is induced by meniscectomy at day 0 (DO) in the right leg of each rat by a transection of the medial collateral ligament and 4 mm of ligament are removed. Internal part of the meniscus is transected vertically into two flaps which are pushed to the front and the back of the synovial cavity. Sham animals undergo only anaesthesia, skin and muscle incision then suture. On day 1, rats are randomly assigned to a treatment group (n=20 per group) according to their body weight, in order to have a homogenous distribution. From C2 to D21, rats are dosed per os (po) once daily (qd) or twice a day (bid) with compounds formulated in methylcellulose (MC) 0.5% or in HPCD 10% pH3.0.
[0779] 4.1.2.2. Steady-State PK Determination (ssPK)
[0780] After at least 7 days of treatment, blood is sampled at 4 time points post administration: 0, 1, 3 and 6 h (and assuming 24 h is equal to the pre-dose sample), in order to determine steady-state plasma exposure.
[0781] 4.1.2.3. Histology
[0782] At sacrifice, the right tibia of each rat is collected and processed for histological analysis. After 48 h of fixation in 4% formaldehyde, tibias are decalcified in Osteosoft for 7 days, and cut into 2 half parts prior to embedding face to face in paraffin. Five series of sections are cut at 200 m intervals, covering about 1.5 mm of the middle part of the bone. One series of slides is stained with Safranin O and light green for morphological evaluation and OARSI scoring. The other series of slides are mounted with DAPI for chondrocyte density measurement.
[0783] The extent of cartilage injury reflecting osteoarthritis in the tibial plateau is evaluated and scored using the OARSI method based on the grading and the staging of cartilage lesion (Pritzker et al, 2006). The OARSI scoring is assessed in a blinded manner by two different readers. For each tibia, one score is attributed as the median of the OARSI score of the 5 sections.
[0784] For statistical analysis, medians of groups are compared with a stratified Kruskal-Wallis test followed by Dunnett multiple comparison post hoc test.
[0785] Significance levels: ns: not statistically significant; *p<0.05; **p<0.01; ***p<0.001 versus MNX-vehicle. Statistical analyses are done on all groups of the studies.
Final Remarks
[0786] It will be appreciated by those skilled in the art that the foregoing descriptions are exemplary and explanatory in nature, and intended to illustrate the invention and its preferred embodiments. Through routine experimentation, an artisan will recognize apparent modifications and variations that may be made without departing from the spirit of the invention. All such modifications coming within the scope of the appended claims are intended to be included therein. Thus, the invention is intended to be defined not by the above description, but by the following claims and their equivalents.
[0787] All publications, including but not limited to patents and patent applications, cited in this specification are herein incorporated by reference as if each individual publication are specifically and individually indicated to be incorporated by reference herein as though fully set forth.
[0788] It should be understood that factors such as the differential cell penetration capacity of the various compounds can contribute to discrepancies between the activity of the compounds in the in vitro biochemical and cellular assays.
[0789] At least some of the chemical names of compound of the invention as given and set forth in this application, may have been generated on an automated basis by use of a commercially available chemical naming software program, and have not been independently verified. Representative programs performing this function include the Lexichem naming tool sold by Open Eye Software, Inc. and the Autonom Software tool sold by MDL, Inc. In the instance where the indicated chemical name and the depicted structure differ, the depicted structure will control.
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