Patent Application
20080269282
1. Field of the Invention
The present invention relates to inhibitors of copper-containing amine oxidases (E.C. 1.4.3.6). Specifically, the invention provides inhibitors of semicarbazide-sensitive amine oxidase (SSAO; also known as Vascular adhesion protein-1 VAP-1). The invention provides methods for using such compounds as therapeutic agents for treating inflammatory diseases, diabetes and its associated complications, atherosclerosis, neurodegenerative diseases, obesity, hypertension and cancer.
2. Background of the Related Art
Semicarbazide-sensitive amine oxidase (SSAO)/Vascular Adhesion Protein-1 (VAP-1) is a bifunctional membrane protein. One function of this protein is as a copper-containing ectoenzyme with amine oxidase activity that can be inhibited by carbonyl-reactive compounds such as semicarbazide (Lyles, 1996, Int. J. Biochem. Cell Biol. 28:259-274). SSAO oxidizes a primary amine into the corresponding aldehyde with production of hydrogen peroxide and ammonia according to the following reaction:
R—CH2—NH2+O2→R—CHO+H2O2+NH3
SSAO/VAP-1 is also an adhesion molecule implicated in inflammation processes (Bono et al., 1999, Amer J Pathol 155:1613-1624; Salmi & Jalkanen, 1992, Science 257:1407-1409; Smith et al., 1998, J. Exp. Med. 188:17-27)
SSAO/VAP-1 is expressed in a variety of tissues, including endothelial cells, lung, smooth muscle cells, and (under normal conditions, highly expressed) in adipose tissue cells. SSAO/VAP-1 is not expressed in 3T3-L1 fibroblasts, but is induced during adipogenesis (Fontana et al., 2001, Biochem. J 356:769-777; Moldes et al., 1999, J. Biol. Chem. 274:9515-9523). This suggests that SSAO/VAP-1 is a member of the adipogenic gene program and, in addition, that SSAO/VAP-1 may contribute to the acquisition of some final characteristics of fully differentiated adipose cells.
SSAO substrates are known to strongly stimulate glucose transport and recruitment of GLUT4 to the cell surface in isolated rat adipocytes or 3T3-L1 adipocytes (Enrique-Tarancon et al., 1998, J. Biol. Chem. 273:8025-8032; Enrique-Tarancon et al., 2000, Biochem. J. 350:171-180; Fontana et al., 2001, Biochem. J 356:769-777; Marti et al., 1998, J Pharmacol. Exp. Ther. 285:342-349). Stimulation of glucose transport by SSAO substrates has also been demonstrated in isolated human adipocytes (Morin et al., 2001, J Pharmacol. Exp. Ther. 297:563-572).
The identity of SSAO and VAP-1 has more recently been established (Bono et al., 1999, Amer J Pathol 155:1613-1624; Smith et al., 1998, J Exp. Med. 188:17-27) VAP-1, first disclosed in Salmi et al. in 1992 (Salmi & Jalkanen, 1992, Science 257:1407-1409) is upregulated (i.e., its expression increases) on the vascular endothelium at inflammation sites, and mediates a multistep adhesive process leading to the transmigration of leukocytes from the circulation into inflamed tissues. Lymphocyte adhesion to endothelial cells is mediated by SSAO/VAP-1 in a sialic acid-dependent manner (Bono et al., 1998, J Immunol. 160:5563-5571) and more recently it has been shown that the SSAO amine oxidase activity of VAP-1/SSAO also participates to the adhesive function of VAP-1 (Salmi et al., 2001, Immunity. 14:265-276).
VAP-1/SSAO has been implicated in a variety of inflammatory responses through its enzymatic activity. These include lymphocyte adhesion (Kurkijarvi et al., 1998, J Immunol. 161:1549-1557; Salmi & Jalkanen, 1992, Science 257:1407-1409; Salmi et al., 2001, Immunity. 14:265-276); and production of aldehydes like formaldehyde or methylglyoxal, putatively endogenous products of SSAO, by generating protein cross-linking and advanced glycosylation end-product (AGE) formation (Yu, 1998, J Neural Transom. Supply 52:201-21). Additionally, VAP-1/SSAO has the capacity to promote LDL oxidation in vitro (Exner et al., 2001, Cardiovasc. Res. 50:583-588) (perhaps through its copper ion), and mice overexpressing VAP-1/SSAO in endothelial cells have a propensity to atherosclerosis (Stolen et al., 2004, FASEB J. 18: 702-704).
VAP-1/SSAO has also been implicated in cardiovascular complications associated with diabetes, adipogenicity, apoptosis secondary to stroke and hypertension. The higher activity of SSAO associated with diabetes as well as a higher concentration of its putative endogenous substrates, methylamine and aminoacetone, may result in greater production of formaldehyde, methylglyoxal and hydrogen peroxide in diabetics than in normal individuals. These products are highly cytotoxic for endothelial cells, which may lead to cardiovascular complications associated to diabetes (Yu, 1998, J Neural Transm. Suppl 52:201-218).
In addition to membrane-bound adhesion molecules, soluble isoforms of SSAO/VAP-1 have been detected in blood plasma from healthy individuals (Gearing & Newman, 1993, Immunol. Today 14:506-512, 1993; Kurkijarvi et al., 1998, J Immunol. 161:1549-1557; Rothlein et al., 1991, J Immunol. 147:3788-3793). The soluble form of VAP-1/SSAO is found in healthy adult plasma at concentrations of 50-140 ng/mL, which is enhanced in inflammatory liver diseases (Kurkijarvi et al., 1998, J Immunol. 161:1549-1557), cardiovascular pathologies (Boomsma et al., 1997, Cardiovasc. Res. 33:387-391), endstage of renal disease (Kurkijarvi et al., 2001, Eur. J Immunol. 31:2876-2884), obesity (Meszaros et al., 1999, Metabolism 48:113-117; Weiss et al., 2003, Metabolism 52:688-692) type I diabetes (Hayes & Clarke, 1990, Res. Commun. Chem. Pathol Pharmacol. 69:71-83; Boomsma et al., 1995, Clin. Sci. (Lond) 88:675-679; Boomsma et al., 1999, Diabetologia 42:233-237; Meszaros et al., 1999, Metabolism 48:113-117; Salmi et al., 2002, Am J Pathol 161:2255-2262) and type II diabetes (Boomsma et al., 1999, Diabetologia 42:233-237; Garpenstrand et al., 1999, Diabet. Med. 16:514-521; Meszaros et al., 1999, Metabolism 48:113-117). The soluble form of VAP-1/SSAO enhances the binding capacity of lymphocytes to endothelial cells (Kurkijarvi et al., 1998, J Immunol. 161:1549-1557) presumably through a lymphocyte preactivation signal. In addition, SSAO aldehyde products, such as formaldehyde or methylglyoxal, may generate protein cross-linking or AGE products implicated in atherogenic lesions, retinopathy and angiopathy associated with diabetes.
Because all these events can promote inflammation and atherosclerosis (Osterud & Bjorklid, 2003, Physiol Rev. 83:1069-1112), pharmacological inhibition of VAP-1/SSAO may reduce a variety of pathologies. A need exists, therefore, for inhibitors of SSAO/VAP-1 that can be used therapeutically to alleviate the symptoms and pathology associated with expression or over-expression of SSAO/VAP-1 in a number of disease states and other disorders.
The present invention provides SSAO/VAP-1 inhibitors having the general formula I:
R3—(Y)n—(CR2R2)m-Z
or a pharmaceutically acceptable salt thereof, wherein
The invention also provides methods for preparing a compound of formula I.
The invention further provides compounds of formula I prepared according to the methods of the invention.
The invention specifically provides methods for inhibiting SSAO/VAP-1 using the compounds of the invention.
The invention also provides pharmaceutical compositions comprising the SSAO/VAP-1 inhibitors of the invention and a pharmaceutically-acceptable diluent, solvent, excipient and/or adjuvant.
The invention further provides methods for treating a disease or disorder associated with SSAO/VAP-1 activity in an animal, wherein said SSAO/VAP-1 activity is inhibited in the animal, preferably by administering to the animal a compound or pharmaceutical composition of the SSAO/VAP-1 inhibitors of the invention. Preferably, the animal is a human.
Specific preferred embodiments of the present invention will become evident from the following more detailed description of certain preferred embodiments and the claims.
As indicated above, the invention provides methods of preparing compounds of formula I, which are inhibitors of copper-containing amine oxidases (E.C. 1.4.3.6) including semicarbazide-sensitive amine oxidase (SSAO; also known as Vascular adhesion protein-1, VAP-1).
In one aspect, the invention provides compounds of formula I-a, i.e., compounds of formula I, or a pharmaceutically-acceptable salt thereof, wherein
In another aspect, the invention provides compounds of formula I-b, i.e., compounds of formula I, or a pharmaceutically-acceptable salt thereof, wherein
In still another aspect, the invention provides compounds of formula I-c, i.e., compound of formula I, or a pharmaceutically-acceptable salt thereof, wherein
In yet another aspect, the invention provides compounds of formula I-d, i.e., compounds of formula I, or a pharmaceutically-acceptable salt thereof, wherein
In another aspect, the invention provides compounds of formula I-e, i.e., compounds of formula I-d, or a pharmaceutically-acceptable salt thereof, wherein
In another aspect, the invention provides compounds of formula I-f, i.e., compounds of formula I-c or I-d, or a pharmaceutically-acceptable salt thereof, wherein Z is CONR1OH, and R1 is H, or C1-C6 alkyl.
In yet another aspect, the invention provides compounds of formula I-g, i.e., compounds of formula I-c or I-d, or a pharmaceutically-acceptable salt thereof, wherein R1 is H.
In still another aspect, the invention provides compounds of formula I-h, i.e., compounds of formula I-c or I-d, or a pharmaceutically-acceptable salt thereof, wherein R1 is C1-C6 alkyl.
In still yet another aspect, the invention provides compounds of formula I-i, i.e., compounds of formula I-c or I-d, or a pharmaceutically-acceptable salt thereof, wherein Z is CONHR1.
In yet still another aspect, the invention provides compounds of formula I-j, i.e., compounds of formula I-c or I-d, wherein R2 is independently H, or C1-C6 alkyl, where the alkyl group is optionally substituted with one or two groups that are independently halogen, C1-C4 alkoxy, phenyl, naphthyl, nitro, CHO, carboxyl, C(O)NH2, C(O)N—H(C1-C6 alkyl), C(O)N(C1-C6 alkyl)(C1-C6 alkyl), NH2, NH(C1-C6 alkyl), N(C1-C6 alkyl)(C1-C6 alkyl), hydroxyl, C1-C6 alkoxycarbonyl, or nitrile.
In another aspect, the invention provides compounds of formula I-k, i.e., compounds of formula I-j, wherein one R2 is H and the other is H or C1-C6 alkyl, where the alkyl group is optionally substituted with OH, NH2, or SH.
In another aspect, the invention provides compounds of formula I-1, i.e., compounds of formula I, I-a, I-b, I-c, I-d, I-e, I-f, I-g, I-h, I-l, I-j, or I-k, wherein Y is —NR2—, —(C1-C6 alkyl)-NHC(O)—, —(C1-C6 alkyl)-N(C1-C6 alkyl)C(O)—, —NHC(O)—, —N(C1-C6alkyl)C(O)—, —C(O)NH—, —C(O)—N(C1-C6alkyl), —SO2NH—, —SO2—N(C1-C6alkyl)-, —(C1-C6 alkyl)-C(O)NH—, or —(C1-C6 alkyl)-C(O)—N(C1-C6alkyl)-, wherein the alkyl portion or portions of each of the above is optionally substituted with 1, 2, or 3 groups that are independently halogen, C1-C4 alkoxy, amino, mono or di (C1-C6 alkyl)amino, OH, indolyl, guanidinyl, carboxyl, or ═O.
In another aspect, the invention provides compounds of formula I-m, i.e., compounds of formula I-1, wherein Y is —NR2—, —NHC(O)—, —C(O)NH—, —SO2NH—, or —(C1-C6 alkyl)-C(O)—N(C1-C6alkyl)-, wherein the alkyl portion or portions of each of the above is optionally substituted with 1, 2, or 3 groups that are independently halogen, C1-C4 alkoxy, amino, mono or di (C1-C6 alkyl)amino, OH, indolyl, guanidinyl, carboxyl, or ═O.
In another aspect, the invention provides compounds of formula I-n, i.e., compounds of formula I-m, wherein Y is —NR2—.
In another aspect, the invention provides compounds of formula I-o, i.e., compounds of formula I-m, wherein Y is —NHC(O)—.
In another aspect, the invention provides compounds of formula I-p, i.e., compounds of formula I-m, wherein Y is —C(O)NH—.
In another aspect, the invention provides compounds of formula I-q, i.e., compounds of formula I-m, wherein Y is —SO2NH—.
In another aspect, the invention provides compounds of formula I-r, i.e., compounds of formula I-m, wherein Y is —(C1-C6 alkyl)-C(O)—N(C1-C6alkyl)-, wherein the alkyl portion or portions of each of the above is optionally substituted with 1, or 2 groups that are independently halogen, C1-C4 alkoxy, amino, mono or di (C1-C6 alkyl)amino, OH, indolyl, guanidinyl, carboxyl, or ═O.
In yet another aspect, the invention provides compounds of formula II, i.e., compounds of formula I-e, of the formula:
wherein Y, R2, Z, n and m are as defined herein with regard to compounds of Formula I.
In another aspect, the invention provides compounds of formula II-a, i.e., compounds of formula II, or a pharmaceutically-acceptable salt thereof, wherein
In yet another aspect, the invention provides compounds of formula II-b, i.e., compounds of formula II-a, wherein n is 1 and m is 1, 2, or 3.
In yet another aspect, the invention provides compounds of formula II-c, i.e., compounds of formula II-b wherein Z is CONR1OH, and R1 is H.
In still yet another aspect, the invention provides compounds of formula II-d, i.e., compounds of formula II-b wherein Z is CONR1OH, and R1 is C1-C4 alkyl.
In still yet another aspect, the invention provides compounds of formula II-e, i.e., compounds according to any of formulas II, II-a, II-b, II-c, or II-d, wherein m is 1 or 2 and at least one R2 is hydrogen.
In yet still another aspect, the invention provides a compound of formula II-f, i.e., compounds according to formula II-e, wherein Z is CONR1OH, R1 is H, and both R2 groups are hydrogen.
In still yet another aspect, the invention provides compounds of formula II-g, i.e., compounds of formula II, or a pharmaceutically-acceptable salt thereof, wherein Z is CONHR1.
In yet still another aspect, the invention provides compounds of formula II-h, i.e., compounds of formula II, II-a, II-b, II-c or II-d, wherein R2 is independently H, or C1-C6 alkyl, where the alkyl group is optionally substituted with one or two groups that are independently halogen, C1-C4 alkoxy, phenyl, naphthyl, halogen, nitro, CHO, carboxyl, C(O)NH2, C(O)NH(C1-C6 alkyl), C(O)N(C1-C6 alkyl)(C1-C6 alkyl), NH2, NH(C1-C6 alkyl), N(C1-C6 alkyl)(C1-C6 alkyl), hydroxyl, C1-C6 alkoxycarbonyl, or nitrile.
In another aspect, the invention provides compounds of formula II, i.e., compounds of formula II-h, wherein one R2 is H and the other is H or C1-C6 alkyl, where the alkyl group is optionally substituted with OH, NH2, or SH.
In another aspect, the invention provides compounds of formula II-j, i.e., compounds of formula II, II-a, II-b, II-c, II-d, II-e, II-f, II-g, II-h, or II-I, wherein Y is —NR2—, —(C1-C6 alkyl)-NHC(O)—, —(C1-C6 alkyl)-N(C1-C6 alkyl)C(O)—, —NHC(O)—, —N(C1-C6alkyl)C(O)—, —C(O)NH—, —C(O)—N(C1-C6alkyl), —SO2NH—, —SO2—N(C1-C6alkyl)-, —(C1-C6 alkyl)-C(O)NH—, or —(C1-C6 alkyl)-C(O)—N(C1-C6alkyl)-, wherein the alkyl portion or portions of each of the above is optionally substituted with 1, 2, or 3 groups that are independently halogen, C1-C4 alkoxy, amino, mono or di (C1-C6 alkyl)amino, OH, indolyl, guanidinyl, carboxyl, or ═O.
In another aspect, the invention provides compounds of formula II-k, i.e., compounds of formula II-j, wherein Y is —NR2—, —NHC(O)—, —C(O)NH—, —SO2NH—, or —(C1-C6 alkyl)-C(O)—N(C1-C6alkyl)-, wherein the alkyl portion or portions of each of the above is optionally substituted with 1, 2, or 3 groups that are independently halogen, C1-C4 alkoxy, amino, mono or di (C1-C6 alkyl)amino, OH, indolyl, guanidinyl, carboxyl, or ═O.
In another aspect, the invention provides compounds of formula II-n, i.e., compounds of formula II-k, wherein Y is —NR2—.
In another aspect, the invention provides compounds of formula II-o, i.e., compounds of formula II-k, wherein Y is —NHC(O)—.
In another aspect, the invention provides compounds of formula II-p, i.e., compounds of formula II-k, wherein Y is —C(O)NH—.
In another aspect, the invention provides compounds of formula II-q, i.e., compounds of formula II-k, wherein Y is —SO2NH—.
In another aspect, the invention provides compounds of formula II-r, i.e., compounds of formula II-k, wherein Y is —(C1-C6 alkyl)-C(O)—N(C1-C6alkyl)-, wherein the alkyl portion or portions of each of the above is optionally substituted with 1, or 2 groups that are independently halogen, C1-C4 alkoxy, amino, mono or di (C1-C6 alkyl)amino, OH, indolyl, guanidinyl, carboxyl, or ═O.
In yet another aspect, the invention provides compounds of formula III, i.e., compounds of formula I-e, of the formula:
wherein Y, R2, Z, n and m are as defined herein with regard to compounds of Formula I
In another aspect, the invention provides compounds of formula III-a, i.e., compounds of formula III, or a pharmaceutically-acceptable salt thereof, wherein
In yet another aspect, the invention provides compounds of formula II-b, i.e., compounds of formula III-a, wherein n is 1 and m is 1, 2, or 3.
In yet another aspect, the invention provides compounds of formula III-c, i.e., compounds of formula II-b wherein Z is CONR1OH, and R1 is H.
In still yet another aspect, the invention provides compounds of formula III-d, i.e., compounds of formula II-b wherein Z is CONR1OH, and R1 is C1-C4 alkyl.
In still yet another aspect, the invention provides compounds of formula III-e, i.e., compounds according to any of formulas III, III-a, III-b, III-c, or III-d, wherein m is 1 or 2 and at least one R2 is hydrogen.
In yet still another aspect, the invention provides a compound of formula II-f, i.e., compounds according to formula III-e, wherein Z is CONR1OH, R1 is H, and both R2 groups are hydrogen.
In still yet another aspect, the invention provides compounds of formula III-g, i.e., compounds of formula III, or a pharmaceutically-acceptable salt thereof, wherein Z is CONHR1.
In yet still another aspect, the invention provides compounds of formula III-h, i.e., compounds of formula III, III-a, III-b, III-c or III-d, wherein R2 is independently H, or C1-C6 alkyl, where the alkyl group is optionally substituted with one or two groups that are independently halogen, C1-C4 alkoxy, phenyl, naphthyl, halogen, nitro, CHO, carboxyl, C(O)NH2, C(O)NH(C1-C6 alkyl), C(O)N(C1-C6 alkyl)(C1-C6 alkyl), NH2, NH(C1-C6 alkyl), N(C1-C6 alkyl)(C1-C6 alkyl), hydroxyl, C1-C6 alkoxycarbonyl, or nitrile.
In another aspect, the invention provides compounds of formula II-i, i.e., compounds of formula II-h, wherein one R2 is H and the other is H or C1-C6 alkyl, where the alkyl group is optionally substituted with OH, NH2, or SH.
In another aspect, the invention provides compounds of formula III-j, i.e., compounds of formula III, III-a, III-b, III-c, III-d, III-e, III-f, III-g, III-h, or III-I, wherein Y is —NR2—, —(C1-C6 alkyl)-NHC(O)—, —(C1-C6 alkyl)-N(C1-C6 alkyl)C(O)—, —NHC(O)—, —N(C1-C6alkyl)C(O)—, —C(O)NH—, —C(O)—N(C1-C6alkyl), —SO2NH—, —SO2—N(C1-C6alkyl)-, —(C1-C6 alkyl)-C(O)NH—, or —(C1-C6 alkyl)-C(O)—N(C1-C6alkyl)-, wherein the alkyl portion or portions of each of the above is optionally substituted with 1, 2, or 3 groups that are independently halogen, C1-C4 alkoxy, amino, mono or di (C1-C6 alkyl)amino, OH, indolyl, guanidinyl, carboxyl, or ═O.
In another aspect, the invention provides compounds of formula III-k, i.e., compounds of formula III-j, wherein Y is —NR2—, —NHC(O)—, —C(O)NH—, —SO2NH—, or —(C1-C6 alkyl)-C(O)—N(C1-C6alkyl)-, wherein the alkyl portion or portions of each of the above is optionally substituted with 1, 2, or 3 groups that are independently halogen, C1-C4 alkoxy, amino, mono or di (C1-C6 alkyl)amino, OH, indolyl, guanidinyl, carboxyl, or ═O.
In another aspect, the invention provides compounds of formula III-n, i.e., compounds of formula III-k, wherein Y is —NR2—.
In another aspect, the invention provides compounds of formula III-o, i.e., compounds of formula III-k, wherein Y is —NHC(O)—.
In another aspect, the invention provides compounds of formula III-p, i.e., compounds of formula III-k, wherein Y is —C(O)NH—.
In another aspect, the invention provides compounds of formula III-q, i.e., compounds of formula III-k, wherein Y is —SO2NH—.
In another aspect, the invention provides compounds of formula III-r, i.e., compounds of formula III-k, wherein Y is —(C1-C6 alkyl)-C(O)—N(C1-C6alkyl)-, wherein the alkyl portion or portions of each of the above is optionally substituted with 1, or 2 groups that are independently halogen, C1-C4 alkoxy, amino, mono or di (C1-C6 alkyl)amino, OH, indolyl, guanidinyl, carboxyl, or ═O.
In yet another aspect, the invention provides compounds of formula IV, i.e., compounds of formula I-e, of the formula:
wherein Y, R2, Z, n and are as defined herein with regard to compounds of Formula I In another aspect, the invention provides compounds of formula IV-a, i.e., compounds of formula IV, or a pharmaceutically-acceptable salt thereof, wherein
In yet another aspect, the invention provides compounds of formula IV-b, i.e., compounds of formula IV-a, wherein n is 1 and m is 1, 2, or 3.
In yet another aspect, the invention provides compounds of formula IV-c, i.e., compounds of formula IV-b wherein Z is CONR1OH, and R1 is H.
In still yet another aspect, the invention provides compounds of formula IV-d, i.e., compounds of formula IV-b wherein Z is CONR1OH, and R1 is C1-C4 alkyl.
In still yet another aspect, the invention provides compounds of formula IV-e, i.e., compounds according to any of formulas IV, IV-a, IV-b, IV-c, or IV-d, wherein m is 1 or 2 and at least one R2 is hydrogen.
In yet still another aspect, the invention provides a compound of formula IV-f, i.e., compounds according to formula IV-e, wherein Z is CONR1OH, R1 is H, and both R2 groups are hydrogen.
In still yet another aspect, the invention provides compounds of formula IV-g, i.e., compounds of formula IV, or a pharmaceutically-acceptable salt thereof, wherein Z is CONHR1.
In yet still another aspect, the invention provides compounds of formula IV-h, i.e., compounds of formula IV, IV-a, IV-b, IV-c or IV-d, wherein R2 is independently H, or C1-C6 alkyl, where the alkyl group is optionally substituted with one or two groups that are independently halogen, C1-C4 alkoxy, phenyl, naphthyl, halogen, nitro, CHO, carboxyl, C(O)NH2, C(O)NH(C1-C6 alkyl), C(O)N(C1-C6 alkyl)(C1-C6 alkyl), NH2, NH(C1-C6 alkyl), N(C1-C6 alkyl)(C1-C6 alkyl), hydroxyl, C1-C6 alkoxycarbonyl, or nitrile.
In another aspect, the invention provides compounds of formula IV-i, i.e., compounds of formula IV-h, wherein one R2 is H and the other is H or C1-C6 alkyl, where the alkyl group is optionally substituted with OH, NH2, or SH.
In another aspect, the invention provides compounds of formula IVj, i.e., compounds of formula IV, IV-a, IV-b, IV-c, IV-d, IV-e, IV-f, IV-g, IV-h, or IV-I, wherein Y is —NR2—, —(C1-C6 alkyl)-NHC(O)—, —(C1-C6 alkyl)-N(C1-C6 alkyl)C(O)—, —NHC(O)—, —N(C1-C6alkyl)C(O)—, —C(O)NH—, —C(O)—N(C1-C6alkyl), —SO2NH—, —SO2—N(C1-C6alkyl)-, —(C1-C6 alkyl)-C(O)NH—, or —(C1-C6 alkyl)-C(O)—N(C1-C6alkyl)-, wherein the alkyl portion or portions of each of the above is optionally substituted with 1, 2, or 3 groups that are independently halogen, C1-C4 alkoxy, amino, mono or di (C1-C6 alkyl)amino, OH, indolyl, guanidinyl, carboxyl, or ═O.
In another aspect, the invention provides compounds of formula IV-k, i.e., compounds of formula IV-j, wherein Y is —NR2—, —NHC(O)—, —C(O)NH—, —SO2NH—, or —(C1-C6 alkyl)-C(O)—N(C1-C6alkyl)-, wherein the alkyl portion or portions of each of the above is optionally substituted with 1, 2, or 3 groups that are independently halogen, C1-C4 alkoxy, amino, mono or di (C1-C6 alkyl)amino, OH, indolyl, guanidinyl, carboxyl, or ═O.
In another aspect, the invention provides compounds of formula IV-n, i.e., compounds of formula IV-k, wherein Y is —NR2—.
In another aspect, the invention provides compounds of formula IV-o, i.e., compounds of formula IV-k, wherein Y is —NHC(O)—.
In another aspect, the invention provides compounds of formula IV-p, i.e., compounds of formula IV-k, wherein Y is —C(O)NH—.
In another aspect, the invention provides compounds of formula IV-q, i.e., compounds of formula IV-k, wherein Y is —SO2NH—.
In another aspect, the invention provides compounds of formula IV-r, i.e., compounds of formula IV-k, wherein Y is —(C1-C6 alkyl)-C(O)—N(C1-C6alkyl)-, wherein the alkyl portion or portions of each of the above is optionally substituted with 1, or 2 groups that are independently halogen, C1-C4 alkoxy, amino, mono or di (C1-C6 alkyl)amino, OH, indolyl, guanidinyl, carboxyl, or ═O.
In certain other aspect, the invention provides compounds of formula B, wherein
wherein n, R3, and Z are as defined in Formula I;
In another aspect, the invention provides compounds of formula B-1, i.e., compounds of formula B wherein R3 is C1-C4 alkyl substituted with phenyl, where the phenyl is optionally substituted with 1, 2, or 3 groups that are independently C1-C6 alkyl (in one aspect, C1-C4 alkyl, in another aspect, C1-C2 alkyl), OH, C1-C6 alkoxy, or phenyl; or
In yet another aspect, the invention provides compounds of formula B-2, i.e., compounds of formula B wherein R3 is C2-C3 alkenyl substituted with furanyl or phenyl, where the phenyl is optionally substituted with 1, 2, or 3 groups that are independently C1-C6 alkyl (in one aspect, C1-C4 alkyl, in another aspect, C1-C2 alkyl), OH, C1-C6 alkoxy, or phenyl.
In still yet another aspect, the invention provides compounds of formula B-3, i.e., compounds of formula B wherein R3 is indolyl, or phenyl, where the phenyl is optionally substituted with 1, 2, or 3 groups that are independently C1-C6 alkyl (in one aspect, C1-C4 alkyl, in another aspect, C1-C2 alkyl), OH, C1-C6 alkoxy, or phenyl.
In yet still another aspect, the invention provides compounds of formula B-4, i.e., compounds of formula B, B-1, B-2, or B-3, wherein Z is —C(O)NHOH.
In certain other aspect, the invention provides compounds of formula C, wherein
wherein Z is as defined in formula I;
R10 is H, amino, mono or di (C1-C6 alkyl)amino, OH, or SH;
R30 is H or C1-C4 alkyl (in one aspect, methyl); and
the C-ring is phenyl or naphthyl, each of which is optionally substituted with one or more groups that are independently OH, NO2, halogen (in one aspect, F), C1-C6 alkyl (in one aspect, methyl), C1-C6 alkoxy (in one aspect, methoxy), amino, mono or di (C1-C6 alkyl)amino, phenyl, phenyl C1-C4 alkoxy (in one aspect, benzyloxy).
In another aspect, the invention provides compounds of formula C-1, i.e., compounds of formula C, wherein the C-ring is phenyl optionally substituted with one or more groups that are independently OH, NO2, halogen (in one aspect, F), C1-C6 alkyl (in one aspect, methyl), C1-C6 alkoxy (in one aspect, methoxy), amino, mono or di (C1-C6 alkyl)amino, phenyl, phenyl C1-C4 alkoxy (in one aspect, benzyloxy).
In another aspect, the invention provides compounds of formula C-2, i.e., compounds of formula C, wherein the C-ring is naphthyl optionally substituted with one or more groups that are independently OH, NO2, halogen (in one aspect, F), C1-C6 alkyl (in one aspect, methyl), C1-C6 alkoxy (in one aspect, methoxy), amino, mono or di (C1-C6 alkyl)amino, phenyl, phenyl C1-C4 alkoxy (in one aspect, benzyloxy).
In another aspect, the invention provides compounds of formula C-3, i.e., compounds of formula C, wherein Z is C(O)NHOH.
In certain other aspect, the invention provides compounds of formula D, wherein
In another aspect, the invention provides compounds of formula D-1, i.e., compounds of formula D wherein R10 is H, C1-C6 alkyl.
In another aspect, the invention provides compounds of formula D-2, i.e., compounds of formula D-1 wherein Y, is OH, NH2, or SH.
In another aspect, the invention provides compounds of formula D-3, i.e., compounds of formula D-2 wherein R20 and R30 are independently H, C1-C6 alkyl, phenyl, C1-C6 alkoxy, NO2, CF3, amino, mono or di (C1-C4 alkyl)amino.
In another aspect, the invention provides compounds of formula D-4, i.e., compounds of formula D-3 wherein the
portion of the molecule is a naphthyl group optionally substituted with R20 and R30.
In another aspect, the invention provides compounds of formula D-5, i.e., compounds of formula D-3 wherein the
portion of the molecule is a quinolinyl group optionally substituted with R20 and R30.
In another aspect, the invention provides compounds of formula D-6, i.e., compounds of formula D, D-1, D-2, D-3, D-4, or D-5, wherein W is NHOH.
In certain other aspect, the invention provides compounds of formula F, wherein
In another aspect, the invention provides compounds of formula F-1, i.e., compounds of formula F, wherein m is 0.
In yet another aspect, the invention provides compounds of formula F-2, i.e., compounds of formula F-1, wherein R30 and R40 are independently H, OH, SH, halogen, nitro, amino, mono or di(C1-C6 alkyl)amino, C1-C6 alkyl, C1-C6 alkanoyl, or C1-C6 alkoxy.
In still another aspect, the invention provides compounds of formula F-3, i.e., compounds of formula F-2, wherein Y1 is (CH2)n, and n is 0, 1, or 2 (in one aspect, 2).
In yet still another aspect, the invention provides compounds of formula F-4, i.e., compounds of formula F-2, wherein Y1 is phenyl, naphthyl, or biphenyl.
In yet still another aspect, the invention provides compounds of formula F-5, i.e., compounds of formula F-3, or F-4, wherein W is NHOH.
In certain other aspect, the invention provides compounds of formula I, wherein
In another aspect, the invention provides compounds of formula I-1, i.e., compounds of formula I, wherein R40 is phenyl, naphthyl, furanyl, indolyl, or quinolinyl, where each of the above is optionally substituted with 1, 2, or 3 groups that are independently halogen, C1-C6 alkyl, C1-C6 alkoxy, phenyl, halogen, nitro, carboxylic acid, —C(O)NH2, —C(O)NH(C1-C6 alkyl), —C(O)N(C1-C6 alkyl)(C1-C6 alkyl), amino, C1-C6 alkoxycarbonyl, hydroxyl or nitrile.
In yet still another aspect, the invention provides compounds of formula I-2, i.e., compounds of formula I-1, wherein n is 1 or 2 and R10 and R20 are both H.
In still yet another aspect, the invention provides compounds of formula I-3, i.e., compounds of formula I-1, wherein n is 0.
In another aspect, the invention provides compounds of formula I-4, i.e., compounds of formula I-1 or 1-2, wherein W is NHOH.
In still yet another aspect, the invention provides compounds of formula I-5, i.e., compounds of formula I-1, wherein R10, R20 and the carbon to which they are attached form a cycloalkyl ring (preferably C5-C6 cycloalkyl) and W is NHOH.
In certain other aspect, the invention provides compounds of formula J, wherein
also encompasses olefins when n is at least 2, in such a case, R20 is not present on the olefinic carbons; or
In another aspect, the invention provides compounds of formula J-1, i.e., compounds of formula J, wherein Y1 is C(O).
In yet another aspect, the invention provides compounds of formula J-2, i.e., compounds of formula J-1, wherein at least one W is OR10 (in one aspect, R10 is H or C1-C6 alkyl; in another aspect, both W groups are OH).
In still another aspect, the invention provides compounds of formula J-3, i.e., compounds of formula J-2, wherein R40 is phenyl, indolyl, or furanyl, where each of the above is optionally substituted with 1, 2, or 3 groups that are independently halogen, C1-C6 alkyl (in one aspect, C1-C2 alkyl), C1-C6 alkoxy (in one aspect, C1-C2 alkoxy), aryl (in one aspect, phenyl), halogen, nitro, carboxylic acid, —C(O)NH2, —C(O)NH(C1-C6 alkyl), —C(O)N(C1-C6 alkyl)(C1-C6 alkyl), amino, C1-C6 alkoxycarbonyl, hydroxyl or nitrile.
In yet still another aspect, the invention provides compounds of formula J-4, i.e., compounds of formula J-3, wherein when n is 2;
is a C2 olefin; R20 is absent and R30 is H or C1-C4 alkyl.
In still another aspect, the invention provides compounds of formula J-5, i.e., compounds of formula J-3, wherein R20, R30, and the carbon to which they are attached form a cycloalkyl ring (in one aspect, a C5 cycloalkyl ring) and at least one W is OH (in another aspect, both W groups are OH).
In certain other aspect, the invention provides compounds of formula K, wherein
In another aspect, the invention provides compounds of formula K-1, i.e., compounds of formula K, wherein Y1 is CO.
In still another aspect, the invention provides compounds of formula K-2, i.e., compounds of formula K-1, wherein
is phenyl, naphthyl, pyridyl, or quinolinyl, each of which is optionally substituted with 1, 2, or 3 groups that are independently OH, SH, C1-C4 alkyl, C1-C4 alkoxy, or phenyl.
In yet still another aspect, the invention provides compounds of formula K-2, i.e., compounds of formula K-1, wherein W is NHOH or NH2.
In certain other aspect, the invention provides compounds of formula M, wherein
In another aspect, the invention provides compounds of formula N-1, i.e., compounds of formula M, wherein R10 is H or C1-C4 alkyl.
In still another aspect, the invention provides compounds of formula N-2, i.e., compounds of formula N-1, wherein R20 is C1-C6 alkyl substituted with phenyl or naphthyl, where the each is optionally substituted with 1 or 2 groups that are independently OH, halogen, C1-C4 alkyl (in one aspect, methyl), or C1-C4 alkoxy (in one aspect, methoxy).
In certain other aspect, the invention provides compounds of formula N, wherein
In another aspect, the invention provides compounds of formula N-1, i.e., compounds of formula N, wherein R10 is H or C1-C4 alkyl.
In still another aspect, the invention provides compounds of formula N2, i.e., compounds of formula N-1, wherein R20 is C1-C6 alkyl substituted with phenyl or naphthyl, where the each is optionally substituted with 1 or 2 groups that are independently OH, halogen, C1-C4 alkyl (in one aspect, methyl), or C1-C4 alkoxy (in one aspect, methoxy).
By “alkyl”, and “C1-C6 alkyl” in the present invention is meant straight or branched chain alkyl groups having 1-6 carbon atoms, such as, methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, pentyl, 2-pentyl, isopentyl, neopentyl, hexyl, 2-hexyl, 3-hexyl, and 3-methylpentyl.
By “alkoxy”, and “C1-C6 alkoxy” in the present invention is meant straight or branched chain alkoxy groups having 1-6 carbon atoms, such as, for example, methoxy, ethoxy, propoxy, isopropoxy, n-butoxy, sec-butoxy, tert-butoxy, pentoxy, 2-pentyl, isopentoxy, neopentoxy, hexoxy, 2-hexoxy, 3-hexoxy, and 3-methylpentoxy.
By the term “halogen” in the present invention is meant fluorine, bromine, chlorine, and iodine.
By “cycloalkyl”, e.g., C3-C7 cycloalkyl, in the present invention is meant cycloalkyl groups having 3-7 atoms such as, for example cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl.
By “aryl” is meant an aromatic carbocyclic group having a single ring (e.g., phenyl), multiple rings (e.g., biphenyl, naphthyl, anthryl, or phenanthryl), or multiple condensed rings in which at least one is aromatic, (e.g., 1,2,3,4-tetrahydronaphthyl), where each aryl group is optionally mono-, di-, or trisubstituted with groups that are independently, e.g., halogen, NO2, amino, NH(C1-C6 alkyl), N(C1-C6 alkyl)(C1-C6 alkyl), C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkylthio, trifluoromethyl, C1-C6 acyloxy, aryl (in one aspect, phenyl), heteroaryl (in one aspect, pyridyl, indolyl, or furanyl), and hydroxy. Preferred aryl groups include phenyl, biphenyl, and naphthyl, each of which is optionally substituted as defined herein. More preferred aryl groups include phenyl and naphthyl, each of which is optionally substituted as defined herein.
By “heteroaryl” is meant an aromatic ring or aromatic ring system, wherein each ring contains of 5-, 6-, or 7-members wherein at least one and up to four ring members are selected from nitrogen, oxygen, or sulfur, and where the heteroaryl group is optionally mono, di, or trisubstituted with groups that are independently, e.g., halogen, NO2, amino, NH(C1-C6 alkyl), N(C1-C6 alkyl)(C1-C6 alkyl), C1-C6 alkyl, C1-C6 alkoxyl C1-C6 alkylthio, trifluoromethyl, C1-C6 acyloxy, aryl (in one aspect, phenyl), heteroaryl (in one aspect, pyridyl, indolyl, or furanyl), and hydroxy. Such heteroaryl groups include, for example, thienyl, furanyl, thiazolyl, imidazolyl, (is)oxazolyl, pyrrolyl, pyrazolyl, imidazolyl, pyridyl, pyrimidinyl, (iso)quinolinyl, indolyl, napthyridinyl, benzimidazolyl, and benzoxazolyl. Preferred heteroaryls are thiazolyl, pyrimidinyl, pyrimidin-2-yl, indolyl, pyridyl, 1-imidazolyl, 2-thienyl, 1-, or 2-quinolinyl, 1-, or 2-isoquinolinyl, 1-, or 2-tetrahydro isoquinolinyl, 2- or 3-furanyl, imidazolyl, and 2-tetrahydrofuranyl.
By “heterocycloalkyl,” is meant one or more carbocyclic ring systems of 3, 4, 5, 6, or 7-membered rings which includes fused ring systems of 9-11 atoms containing at least one and up to four heteroatoms selected from nitrogen, oxygen, and sulfur, and where each heterocycloalkyl group is where each aryl group is optionally mono-, di-, or trisubstituted with groups that are independently, e.g., halogen, NO2, amino, NH(C1-C6 alkyl), N(C1-C6 alkyl)(C1-C6 alkyl), C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkylthio, trifluoromethyl, C1-C6 acyloxy, aryl (in one aspect, phenyl), heteroaryl (in one aspect, pyridyl, indolyl, or furanyl), and hydroxy. Preferred heterocycles of the present invention include morpholinyl, thiomorpholinyl, thiomorpholinyl S-oxide, thiomorpholinyl S,S-dioxide, piperazinyl, homopiperazinyl, pyrrolidinyl, pyrrolinyl, tetrahydropyranyl, piperidinyl, tetrahydrofuranyl, tetrahydrothienyl, homopiperidinyl, homomorpholinyl, homothiomorpholinyl, homothiomorpholinyl S,S-dioxide, oxazolidinonyl, dihydropyrazolyl, dihydropyrrolyl, dihydropyrazinyl, dihydropyridinyl, dihydropyrimidinyl, dihydrofuryl, dihydropyranyl, azepanyl, diazepanyl, tetrahydrothienyl S-oxide, tetrahydrothienyl S,S-dioxide and homothiomorpholinyl S-oxide.
Compounds of formula I are useful as pharmaceutical agents, and can be provided as pharmaceutical compositions. The pharmaceutical compositions can be manufactured in a manner that is itself known, e.g., by means of a conventional mixing, dissolving, granulating, dragee-making, levigating, emulsifying, encapsulating, entrapping or lyophilizing processes.
Pharmaceutical compositions can be formulated in conventional manner using one or more physiologically acceptable carriers comprising excipients and auxiliaries that facilitate processing of the active compounds into preparations that can be used pharmaceutically. Proper formulation is dependent upon the route of administration chosen.
Non-toxic pharmaceutical salts include salts of acids such as hydrochloric, phosphoric, hydrobromic, sulfuric, sulfinic, formic, toluenesulfonic, methanesulfonic, nitic, benzoic, citric, tartaric, maleic, hydroiodic-, alkanoic such as acetic, HOOC—(CH2)n—CH3 where n is 0-4, and the like. Non-toxic pharmaceutical base addition salts include salts of bases such as sodium, potassium, calcium, ammonium, and the like. Those skilled in the art will recognize a wide variety of non-toxic pharmaceutically acceptable addition salts.
For injection, the compounds prepared according to the methods of the invention can be formulated in appropriate aqueous solutions, such as physiologically compatible buffers such as Hanks's solution, Ringer's solution, or physiological saline buffer. For transmucosal and transcutaneous administration, penetrants appropriate to the barrier to be permeated are used in the formulation. Such penetrants are generally known in the art.
For oral administration, the compounds can be formulated readily by combining the active compounds with pharmaceutically acceptable carriers well-known in the art. Such carriers enable the compounds of the invention to be formulated as tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions and the like, for oral ingestion by a patient to be treated. Pharmaceutical preparations for oral use can be obtained with solid excipient, optionally grinding a resulting mixture, and processing the mixture of granules, after adding suitable auxiliaries, if desired, to obtain tablets or dragee cores. Suitable excipients are, in particular, fillers such as sugars, including lactose, sucrose, mannitol, or sorbitol; cellulose preparations such as, for example, maize starch, wheat starch, rice starch, potato starch, gelatin, gum tragacanth, methyl cellulose, hydroxypropylmethylcellulose, sodium carboxymethylcellulose, and/or polyvinylpyrrolidone (PVP). If desired, disintegrating agents can be added, such as the cross-linked polyvinyl pyrrolidone, agar, or alginic acid or a salt thereof such as sodium alginate.
Dragee cores are provided with suitable coatings. For this purpose, concentrated sugar solutions can be used, which can optionally contain gum arabic, talc, polyvinyl pyrrolidone, carbopol gel, polyethylene glycol, and/or titanium dioxide, lacquer solutions, and suitable organic solvents or solvent mixtures. Dyestuffs or pigments can be added to the tablets or dragee coatings for identification or to characterize different combinations of active compound doses.
Pharmaceutical preparations that can be used orally include push-fit capsules made of gelatin, as well as soft, sealed capsules made of gelatin and a plasticizer, such as glycerol or sorbitol. The push-fit capsules can contain the active ingredients in admixture with filler such as lactose, binders such as starches, and/or lubricants such as talc or magnesium stearate and, optionally, stabilizers. In soft capsules, the active compounds can be dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin, or liquid polyethylene glycols. In addition, stabilizers can be added. All formulations for oral administration should be in dosages suitable for such administration. For buccal administration, the compositions can take the form of tablets or lozenges formulated in conventional manner.
For administration by inhalation, the compounds prepared according to the methods of the invention are conveniently delivered in the form of an aerosol spray presentation from pressurized packs or a nebuliser, with the use of a suitable propellant, e.g., dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas. In the case of a pressurized aerosol the dosage unit can be determined by providing a valve to deliver a metered amount. Capsules and cartridges of e.g., gelatin for use in an inhaler or insufflator can be formulated containing a powder mix of the compound and a suitable powder base such as lactose or starch.
The compounds can be formulated for parenteral administration by injection, e.g., by bolus injection or continuous infusion. Formulations for injection can be presented in unit dosage form, e.g., in ampoules or in multi-dose containers, with an added preservative. The compositions can take such forms as suspensions, solutions or emulsions in oily or aqueous vehicles, and can contain formulatory agents such as suspending, stabilizing and/or dispersing agents.
Pharmaceutical formulations for parenteral administration include aqueous solutions of the active compounds in water-soluble form. Additionally, suspensions of the active compounds can be prepared as appropriate oily injection suspensions. Suitable lipophilic solvents or vehicles include fatty oils such as sesame oil, or synthetic fatty acid esters, such as ethyloleate or triglycerides, or liposomes. Aqueous injection suspensions can contain substances that increase the viscosity of the suspension, such as sodium carboxymethyl cellulose, sorbitol, or dextran. Optionally, the suspension can also contain suitable stabilizers or agents that increase the solubility of the compounds to allow for the preparation of highly concentrated solutions. Alternatively, the active ingredient can be in powder form for constitution with a suitable vehicle, e.g., sterile pyrogen-free water, before use. The compounds can also be formulated in rectal compositions such as suppositories or retention enemas, e.g., containing conventional suppository bases such as cocoa butter or other glycerides.
In addition to the formulations described previously, the compounds can also be formulated as a depot preparation. Such long acting formulations can be administered by implantation (for example subcutaneously or intramuscularly) or by intramuscular injection. Thus, for example, the compounds can be formulated with suitable polymeric or hydrophobic materials (for example as an emulsion in acceptable oil) or ion exchange resins, or as sparingly soluble derivatives, for example, as a sparingly soluble salt.
A pharmaceutical carrier for hydrophobic compounds of formula I is a cosolvent system comprising benzyl alcohol, a nonpolar surfactant, a water-miscible organic polymer, and an aqueous phase. The cosolvent system can be the VPD co-solvent system. VPD is a solution of 3% w/v benzyl alcohol, 8% w/v of the nonpolar surfactant polysorbate 80, and 65% w/v polyethylene glycoL300, made up to volume in absolute ethanol. The VPD co-solvent system (VPD:5W) consists of VPD diluted 1:1 with a 5% dextrose in water solution. This co-solvent system dissolves hydrophobic compounds well, and itself produces low toxicity upon systemic administration. Naturally, the proportions of a co-solvent system can be varied considerably without destroying its solubility and toxicity characteristics. Furthermore, the identity of the co-solvent components can be varied: for example, other low-toxicity nonpolar surfactants can be used instead of polysorbate 80; the fraction size of polyethylene glycol can be varied; other biocompatible polymers can replace polyethylene glycol, e.g. polyvinyl pyrrolidone; and other sugars or polysaccharides can substitute for dextrose.
Alternatively, other delivery systems for hydrophobic pharmaceutical compounds can be employed. Liposomes and emulsions are well known examples of delivery vehicles or carriers for hydrophobic drugs. Certain organic solvents such as dimethylsulfoxide also can be employed, although usually at the cost of greater toxicity. Additionally, the compounds can be delivered using a sustained-release system, such as semipermeable matrices of solid hydrophobic polymers containing the therapeutic agent. Various sustained-release materials have been established and are well known by those skilled in the art. Sustained-release capsules can, depending on their chemical nature, release the compounds for a few weeks up to over 100 days. Depending on the chemical nature and the biological stability of the therapeutic reagent, additional strategies for protein and nucleic acid stabilization can be employed.
The pharmaceutical compositions also can comprise suitable solid or gel phase carriers or excipients. Examples of such carriers or excipients include but are not limited to calcium carbonate, calcium phosphate, various sugars, starches, cellulose derivatives, gelatin, and polymers such as polyethylene glycols.
The compounds of Formula I can be provided as salts with pharmaceutically compatible counterions. Pharmaceutically compatible salts can be formed with many acids, including but not limited to hydrochloric, sulfuric, acetic, lactic, tartaric, malic, succinic, phosphoric, hydrobromic, sulfinic, formic, toluenesulfonic, methanesulfonic, nitic, benzoic, citric, tartaric, maleic, hydroiodic, alkanoic such as acetic, HOOC—(CH2)n—CH3 where n is 0-4, and the like. Salts tend to be more soluble in aqueous or other protonic solvents that are the corresponding free base forms. Non-toxic pharmaceutical base addition salts include salts of bases such as sodium, potassium, calcium, ammonium, and the like. Those skilled in the art will recognize a wide variety of non-toxic pharmaceutically acceptable addition salts.
Pharmaceutical compositions of the compounds prepared according to the methods of the invention can be formulated and administered through a variety of means, including systemic, localized, or topical administration. Techniques for formulation and administration can be found in “Remington's Pharmaceutical Sciences,” Mack Publishing Co., Easton, Pa. The mode of administration can be selected to maximize delivery to a desired target site in the body. Suitable routes of administration can, for example, include oral, rectal, transmucosal, transcutaneous, or intestinal administration; potential delivery, including intramuscular, subcutaneous, intramedullary injections, as well as intrathecal, direct intraventricular, intravenous, intraperitoneal, intranasal, or intraocular injections.
Alternatively, one can administer the compound in a local rather than systemic manner, for example, via injection of the compound directly into a specific tissue, often in a depot or sustained release formulation.
Pharmaceutical compositions suitable for use include compositions wherein the active ingredients are contained in an effective amount to achieve its intended purpose. More specifically, a therapeutically effective amount means an amount effective to prevent development of or to alleviate the existing symptoms of the subject being treated. Determination of the effective amounts is well within the capability of those skilled in the art, especially in light of the detailed disclosure provided herein.
For administration to non-human animals, the drug or a pharmaceutical composition containing the drug may also be added to the animal feed or drinking water. It will be convenient to formulate animal feed and drinking water products with a predetermined dose of the drug so that the animal takes in an appropriate quantity of the drug along with its diet. It will also be convenient to add a premix containing the drug to the feed or drinking water approximately immediately prior to consumption by the animal.
Preferred compounds prepared according to the methods of the invention will have certain pharmacological properties. Such properties include, but are not limited to oral bioavailability, low toxicity, low serum protein binding and desirable in vitro and in vivo half-lives. Assays may be used to predict these desirable pharmacological properties. Assays used to predict bioavailability include transport across human intestinal cell monolayers, including Caco-2 cell monolayers. Serum protein binding may be predicted from albumin binding assays. Such assays are described in a review by Oraveova et al. (1996, Journal of Chromatography B-Biomedical Applications 677:1-28). Compound half-life is inversely proportional to the frequency of dosage of a compound. In vitro half-lives of compounds may be predicted from assays of microsomal half-life as described by Kuhnz and Gieschen (1998, Drug Metabolism and Disposition 26:1120-1127).
Toxicity and therapeutic efficacy of such compounds can be determined by conventional pharmaceutical procedures in cell cultures or experimental animals, e.g., for determining the LD50 (the dose lethal to 50% of the population) and the ED50 (the dose therapeutically effective in 50% of the population). The dose ratio between toxic and therapeutic effects is the therapeutic index and it can be expressed as the ratio between LD50 and ED50. Compounds that exhibit high therapeutic indices are preferred. The data obtained from these cell culture assays and animal studies can be used in formulating a range of dosage for use in humans. The dosage of such compounds lies preferably within a range of circulating concentrations that include the ED50 with little or no toxicity. The dosage can vary within this range depending upon the dosage form employed and the route of administration utilized. The exact formulation, route of administration and dosage can be chosen by the individual physician in view of the patient's condition. (See, e.g. Fing et al, 1975, in T
Dosage amount and interval can be adjusted individually to provide plasma levels of the active moiety that are sufficient to maintain bacterial cell growth-inhibitory effects. Usual patient dosages for systemic administration range from 100-2000 mg/day. Stated in terms of patient body surface areas, usual dosages range from 50-910 mg/m2/day. Usual average plasma levels should be maintained within 0.1-1000 μM. In cases of local administration or selective uptake, the effective local concentration of the compound cannot be related to plasma concentration.
Compounds provided by the present invention are useful the treatment or prevention of a plurality of diseases and disorders caused by or associated with SSAO activity or inappropriate activity or expression thereof. Particular disorders include inflammatory diseases, adipocyte dysfunction related diseases, carbohydrate metabolism related diseases, vascular diseases, neurodegenerative diseases or cancer. Said diseases and disorders include but are not limited to inflammatory disease including rheumatoid arthritis, osteoarthritis, spondylitis, bone resorption diseases, sepsis, septic shock, atherosclerosis, retinopathy, including diabetic retinopathy, diabetes, chronic pulmonary inflammatory disease, fever, periodontal diseases, ulcerative colitis, pyresis, Alzheimer's and Parkinson's diseases, cystic fibrosis, dysfunctions of the immune system, diabetes onset and maintenance of pancreatic function in diabetes. Preferred diseases and disorders include stroke, multiple sclerosis, migraine, cancer, pain. Generally, the compounds of the invention are provided to advantageously be used for treating or preventing inflammatory eye conditions including uveitis, glaucoma and conjunctivitis; degenerative bone or joint conditions including osteoarthritis, rheumatoid arthritis, rheumatoid spondylitis, gouty arthritis ankylosing spondylitis, psoriatic arthritis and other arthritic conditions, as well as inflamed joints; chronic inflammatory skin conditions, including allergic lesions, lichen planus, pityriasis rosea, eczema, psoriasis, and dermatitis; diseases and disorders of the gastrointestinal tract, including inflammatory bowel disease, Crohn's disease, atrophic gastritis, gastritis varialoforme, ulcerative colitis, coeliac disease, regional ileitis, peptic ulceration, particularly irritable bowel syndrome, reflux oesophagitis, and damage to the gastrointestinal tract resulting from infections, for example, by Helicobacter pylori, or from treatments with non-steroidal anti-inflammatory drugs; inflammatory lung disorders such as asthma, bronchitis, particularly chronic obstructive pulmonary disease, farmer's lung, acute respiratory distress syndrome; bacteraemia, endotoxaemia (septic shock), aphthous ulcers, gingivitis, pyresis, particularly pain, including inflammatory pain, neuropathic pain, acute pain or pain of a central origin; meningitis and pancreatitis; and other conditions associated with inflammation; central nervous system inflammatory conditions and diseases, including multiple sclerosis, Alzheimer's disease, and ischaemia-reperfusion injury associated with ischemic stroke; vascular diseases, such as atheromatous and nonatheromatous arteriosclerosis, ischemic heart disease, and Raynaud's Disease and Phenomenon; diabetes and its complications such as microvascular and macrovascular diseases such as atherosclerosis, vascular retinopathies, nephropathies and neuropathies.
Prophylaxis is expected to be particularly relevant to the treatment of persons who have suffered a previous episode of, or are otherwise considered to be at increased risk of, the disease or condition in question. Persons at risk of developing a particular disease or condition generally include those having a family history of the disease or condition, or those who have been identified by genetic testing or screening to be particularly susceptible to developing the disease or condition.
As used herein, the terms “treating” or “treatment” of a disease includes: (1) preventing the disease, i.e. causing the clinical symptoms of the disease not to develop in a mammal that may be exposed to or predisposed to the disease but does not yet experience or display symptoms of the disease, (2) inhibiting the disease, i.e., arresting or reducing the development of the disease or its clinical symptoms, or (3) relieving the disease, i.e., causing regression of the disease or its clinical symptoms.
As used herein, the term “therapeutically effective amount” means the amount of a compound that, when administered to a mammal for treating a disease, is sufficient to effect such treatment for the disease. The “therapeutically effective amount” will vary depending on the compound, the disease and its severity and the age, weight, or other relevant characteristics of the mammal to be treated.
The compounds of the present invention may be prepared by use of known chemical reactions and procedures. Representative methods for synthesizing compounds of the invention are presented below. It is understood that the nature of the substituents required for the desired target compound often determines the preferred method of synthesis. All variable groups of these methods are as described in the generic description if they are not specifically defined below.
A first general method (Method I) for preparing the agents of formula (I) is summarized in Reaction Scheme 1.
wherein R1 can be H, C1-C6 alkyl, aryl/or substituted aryl, or cycloalkyl, /or where each cycloalkyl group has from 3-7 members, where up to two of the cycloalkyl/members are optionally hetero atoms selected from sulfur, oxygen and nitrogen, and where any member of the alkyl, aryl/or cycloalkyl group is optionally substituted with halogen, C1-C6 alkyl or C1-C6 alkoxy, aryl or substituted aryl;
wherein R2 can be H, C1-C6 alkyl, aryl/or substituted aryl, C1-C6 alkoxyalkyl, or cycloalkyl or cycloalkyl/alkoxy, where each cycloalkyl has from 3-7 members, where up to two of the cycloalkyl members are optionally hetero atoms selected from sulfur, oxygen and nitrogen, and where any member of the alkyl, aryl or cycloalkyl is optionally substituted with halogen, C1-C6 alkyl/or C1-C6 alkoxy, aryl, halogen, nitro, nitroso, aldehyde, carboxylic acid, —C(O)NH2, —C(O)NH(C1-C6 alkyl), —C(O)N(C1-C6 alkyl)(C1-C6 alkyl), C1-C6 alkoxycarbonyl, sulfate, phosphate, boronic acid, thio, oxime, or imino. Where R2 is aryl or aryl fused ring (where up to two of the cycloalkyl members are optionally hetero atoms selected from sulfur, oxygen and nitrogen), R2 can be further substituted in any position with halogen, nitro, nitroso, aldehyde, carboxylic acid, —C(O)NU2, —C(O)NH(C1-C6 alkyl), —C(O)N(C1-C6 alkyl)(C1-C6 alkyl), amino, C1-C6 alkoxycarbonyl, sulfate, phosphate, boronic acid, thio, oxime, imino, or hydroxyl; and
R3 and Z are as defined in formula I.
This reaction scheme comprises carrying out a condensation with a hydroxylamine group anchored onto a polymeric support (Z=NHR1O), or Rink Amide (Z=NH2), or Wang resin (Z=OH), or BAL resin (Z=NHR1) including the following steps:
a) acylating with the corresponding acid (4 equiv.) the free amino function of the polymeric support using 4 equiv of the corresponding acylating mixture (e.g. HOBt/DIPCDI, HOAt/HATU/DIEA) in 1 mL of DMF at room temperature for 1 h; and
b) releasing the compound of the polymeric support using 1 mL TFA in DCM.
A second general method (Method II) for preparing the agents of formula (I) is summarized in Reaction Scheme 2.
wherein
The method of scheme 2 comprises carrying out a condensation with a hydroxylamine group anchored onto a polymeric support (Z=NHR10), or Rink Amide (Z=NH2), or Wang resin (Z=OH), or BAL resin (Z=NHR1) including the following steps:
a) coupling the amino acid (4 equiv) with the free amino function of the polymeric support using 4 equiv of the corresponding acylating mixture (e.g. HOBt/DIPCDI, HOAt/HATU) in 1 mL of DMF at room temperature for 1 h;
b) removing the protecting group (e.g., 9-fluorenylmethoxycarbonyl) with two treatments of 15 min with 1 ml of piperidine/DMF (50:50) at room temperature;
c1) for sulfonylation: reacting 5 equiv of the corresponding sulfonyl chloride in 1 mL of DCM and 5 equiv. of DIEA at room temperature for 12 h with the free amino function of the amino acid;
c2) for acylation: reacting 3 equiv of the corresponding acid with 3 equiv of HOBt/DIPCDI in 1 mL of DMF as acylating mixture at room temperature for 2 h with the free amino function of the amino acid;
c3) for reductive amination: reacting 5 equiv of the corresponding aldehyde and 5 equiv of NaBH3CN in 1 mL of AcOH/DMF (1:99) for 3 h at room temperature with the free amino function of the amino acid;
c4) for urea formation (two steps procedure): (a) carbamate formation by reacting 5 equiv of 4-nitrophenil chloroformate and 5 equiv of DIEA in 1 mL of DCM for 12 h at 60° C. with the free amino function of the amino acid, and (b) urea formation by reacting 5 equiv of amine with 5 equiv of TEA in 1 mL of NMP for 12 h at 60° C.
d) releasing the compound of the polymeric support in acidic conditions using 1 mL of TFA in DCM.
A third general method (Method III) for preparing the agents of formula (I) is summarized in Reaction Scheme 3.
wherein
wherein n is an integer between 1-6;
The method of Scheme 3 comprises carrying out a condensation with a hydroxylamine group anchored onto a polymeric support (Z=NHR10), or Rink Amide (Z=NH2), or Wang resin (Z=OH), or BAL resin (Z=NHR1) including the following steps:
a) overnight acylation of the free amino function of the polymeric support with 5 equiv of the corresponding anhydride in 1 mL of THF at 60° C.;
b) free acid activation with 25 equiv of CDI in 1 mL of DMF for 30 min;
c) coupling the corresponding amine with 3 equiv of HOBt in 1 mL of DMF for 2 h at room temperature; and
d) releasing the compound from the polymeric support in acidic conditions using 1 mL of % TFA in DCM.
A fourth general method (Method IV) for preparing the agents of formula (I) in solution phase is summarized in Reaction Scheme IV.
wherein
wherein n is an integer between 1-6;
The method of Scheme 4 comprises carrying out a condensation between an amine and carboxylic acid in solution phase, including the following steps:
a) reacting 1 equiv of the corresponding acid with 3 equiv of R3NH2/HOBt/DIPCDI in 1.5 mL of DMF as acylating mixture at room temperature for 2 h at room temperature;
b) drying and purification by normal phase, ISOLUTE HM-N 3.0 cartridge or DIAION HP-20;
b) releasing the protecting group following standard procedures (e.g. acidic conditions using 1 mL of HCl/Dioxane 4 M for Boc protecting groups);
The following abbreviations are used herein:
ACN, acetonitrile;
Alloc, allyloxycarbonyl
Boc, t-butyloxycarbonyl
Bz, benzyl
TFA, trifluoroacetic acid;
THF, tetrahydrofurane;
MeOH, methanol;
F-moc, 9-fluorenylmethyloxycarbonyl;
DMF, dimethylformamide;
DCM, methylenechlorode;
CDI, 1,1′-carbonyldiimidazole;
HOBt, 1-hydroxybenzotriazole;
HOAt, 1-Hydroxy-7-azabentriazole;
DIPCDI, N,N′-diisopropylcarbodiimide;
HATU, (N-dimethylamino)-1H-1,2,3-triazolo(4,5-b)pyridine-1-ylmethylene)-N-ethylmethanominium hexafluorophosphate N-oxide;
Cl-Trt, chlorotrityl resin
ESI-MS, Electrospray ionization mass spectroscopy;
IR, infrared spectroscopy;
HPLC, high performance liquid chromatography;
tR, retention time;
NMR, nuclear magnetic resonance;
LG; leaving group;
Solid-phase manipulations were performed in polypropylene syringes fitted with a polyethylene porous disc. Solvents and soluble reagents were removed by filtration.
Representative compounds prepared according to the methods of the present invention include, but are not limited to the compounds disclosed herein and their pharmaceutically acceptable acid and base addition salts. In addition, if a compound is obtained as an acid addition salt, the free base can be obtained by basifying a solution of the acid salt. Conversely, if the product is a free base, an addition salt, particularly a pharmaceutically acceptable addition salt, may be produced by dissolving the free base in a suitable organic solvent and treating the solution with an acid, in accordance with conventional procedures for preparing acid addition salts from base compounds.
The disclosures in this application of all articles and references, including patents, are incorporated herein by reference.
The following Examples are provided for the purposes of illustration and are not intended to limit the scope of the present invention. The present invention is not to be limited in scope by the exemplified embodiments, which are intended as illustrations of individual aspects of the invention. Indeed, various modifications of the invention in addition to those shown and described herein will become apparent to those skilled in the art from the foregoing description and accompanying drawings. Such modifications are intended to fall within the scope of the appended claims.
General Experimental Procedure. Hydroxylamine Cl-Trt resin (100 mg, 0.8 mmol/gr) was acylated with carboxylic acid (4 equiv) and 4 equi. of the acylating mixture HATU/HOBT/DIEA (1:1:2) in DMF (1 mL) as for 1 h at room temperature. The resin was filtered off and washed with 1 mL of DMF (5×1 min) and 1 mL of DCM (5×1 min). The corresponding hydroxamic acid was cleaved with 1 mL 5% of TFA in DCM (3×1 min) and dried. The crude material was dissolved in water and was purified using Diaion HP-20 (500 mg) following standard procedure. HPLC analyses were performed using the following eluent solutions: H2O (0.1% HCOOH)/ACN(0.07% HCOOH) in a gradient from 0%-100% ACN over 10 min., using an X-Terra C18 5 μm column (4.6×100) and interrogating the column fractions spectrophotometrically (λ=220/254 nm).
(a) 500 mg of DIAION HP-20 in polypropylene syringes fitted with a polyethylene porous disc were conditioned with three fractions of 10 mL MeOH and with three fractions of 10 mL water;
(b) the crude material was dissolved in 5 mL water and the aqueous fraction was eluted trough the resin;
(c) the resin was washed with 3 fractions of 10 mL water;
(d) the product was eluted with three fractions of 10 mL ACN and the solvent was evaporated.
General Experimental Procedure. Hydroxylamine Cl-Trt resin (100 mg, 0.8 mmol/gr) was acylated with Fmoc amino acid (4 equivalents) and 4 equiv. of the acylating mixture HATU/HOBT/DIEA (1:1:2) in 1 mL of DMF for 1 h at room temperature. The Fmoc group was then removed by treating with 1 mL of 50% piperidine in DMF (three times for 10 min apiece), and the free amine was sulfonylated overnight with biphenylsulfonyl chloride (5 equiv) in 1 mL of DCM. The resin was filtered off and washed with 1 mL of DCM (5 washes for 1 min apiece) and the extent of the reaction was checked by the ninhydrin test. The corresponding product was cleaved with 1 mL of 5% TFA in DCM (3 treatments for 1 min apiece) and dried. The crude material was dissolved in water and was purified using Diaion HP-20 (500 mg) following standard procedures as disclosed above. The acetonitrile fraction was dried under vacuum. HPLC analyses were performed using the following eluent solutions: H2O (0.1% HCOOH)/ACN(0.07% HCOOH) in a gradient from 0%-100% ACNover 10 min., using an X-Terra C18 5 μm column (4.6×100) and interrogating the column fractions spectrophotometrically (k=220/254 nm).
General Experimental Procedure. Hydroxylamine Cl-Trt resin (100 mg, 0.8 mmol/gr) was acylated with Fmoc amino acid (4 equiv.) and 4 equiv. of the acylating mixture HATU/HOBT/DIEA (1:1:2) in 1 mL of DMF for 1 h at room temperature. The Fmoc group was then removed by treating with 50% piperidine in 1 mL of DMF (10×3 min.), and the free amine was acylated with the corresponding carboxylic acid (3 equiv.) and 3 equiv. of the acylating mixture HOBT/DIPCDI (1:1) in 1 mL of DMF for 2 h at room temperature. The resin was filtered off and washed with 1 mL of DCM (5×1 min) and the extent of the reaction was determined using the ninhydrin test (Kaiser et al., 1970, Analytical Biochem., 34:595-598). The corresponding product was cleaved with 1 mL of 5% TFA in DCM (3×1 min) and dried. The crude material was dissolved in water and was purified using Diaion HP-20 (500 mg) following standard procedures as disclosed above. The acetonitrile fraction was dried under vacuum. HPLC analyses were performed using the following eluent solutions: H2O (0.1% HCOOH)/ACN(0.07% HCOOH) in a gradient from 0%-100% ACN over 10 min., using an X-Terra C18 5 μm column (4.6×100) and interrogating the column fractions spectrophotometrically (λ=220/254 nm).
General Experimental Procedure. Hydroxylamine Cl-Trt resin (100 mg, 0.8 mmol/gr) was acylated overnight with succinic anhydride (5 equiv) in THF at 60° C. The resin was filtered off and washed with 1 mL of THF (5×1 min) and 1 mL of DCM (5×1 min) and the extent of the reaction was determined by the ninhydrin test as disclosed above. The carboxyl group was then activated with DIC (25 equiv) in 1 mL of DMF for 30 min. at 25° C. The resin was filtered off and washed with 1 mL of DMF (5×1 min). Afterwards, the corresponding amine (3 equiv) and HOBt (3 equiv) in 1 mL of DMF were added and the mixture was stirred for 2 h at room temperature. The resin was filtered off and washed with 1 mL of THF (5×1 min) and 1 mL of DCM (5×1 min), and the extent of the reaction was checked by the malachite green test (Attardi et al., 2000, Tetrahedron Lett., 41: 7391-7394). The corresponding succinic derivative was cleaved with 1 mL of 5% TFA in DC (3×1 min) and dried. The crude material was dissolved in water and was purified using Diaion HP-20 (500 mg) following standard procedures as described above. The acetonitrile fraction was dried under vacuum. HPLC analyses, which were performed using the following eluent solutions: H2O (0.1% HCOOH)/ACN(0.07% HCOOH) in a gradient from 0%-100% ACN over 10 min., using an X-Terra C18 5 μm column (4.6×100) and interrogating the column fractions spectrophotometrically (λ=220/254 nm).
A mixture of 4-tert-Butoxycarbonylamino-3-hydroxy-benzoic acid (1 equiv), HOBT (1 equiv), 1 equiv of the corresponding amine R3NH2 and 1 equiv DIPCDI was stirring in DMF (1.5 mL) overnight at room temperature. DMF was eliminated at low pressure and the crude and was purified using the ISOLUTE HM-N 3.0 cartridge and DIAION HP-20 following the standard procedures. Boc protecting group was removal with 1 mL of HCl/Dioxane 4.0 M treatment for a 1 h at room temperature. HPLC analyses were performed using the following eluent solutions: H2O (0.1% HCOOH)/ACN(0.07% HCOOH) in a gradient from 0%-100% ACN over 10 min., using an X-Terra C18 5 μm column (4.6×100) and interrogating the column fractions spectrophotometrically (λ=220/254 nm).
(a) crude material was dissolved in 1.5 mL of 5% NaHCO3 and 1.5 mL of ethyl acetate and was eluted trough the cartridge;
(b) cartridge was washed with 3 fractions of 4 mL of ethyl acetate the solvent was evaporated.
The following compounds are prepared essentially according to the methods and procedures described above.
The above names were generated using ChemDraw Ultra v. 9.0.1, which is available from CambridgeSoft.
SSAO activity determination: All assays were performed at 37° C. with SSAO from human or mice adipose tissue. The enzyme activity was measured through detection of hydrogen peroxide formed by the oxidation of benzylamine. This method is based on the horseradish peroxidase catalyzed hydrogen peroxide oxidation of 10-acetyl-3,7-dihydroxyphenoxazine (Molecular Probes A-6550), that produces resorufin a highly fluorescent product (excitation, 545 nm; emission, 590 nm) (Zhou and Panchuk-Voloshina, 1997). Human or mice adipose tissue homogenates, used as a source of SSAO activity, were preincubated in 96 well microplates for 20 min at 37° C. in 180 μL of 200 mM Phosphate buffer and H2O2-detecting mixture containing horseradish peroxidase (final concentration 1 U/mL) and Amplex Red reagent (60 μM) and different concentrations of inhibitors when necessary. Catalytic reaction was initiated by addition of 20 μL of benzylamine as substrate at 10 mM for human homogenates giving final concentrations of 100 μM and 1 mM respectively. Fluorescence intensity of the samples was measured continuously during 1 h (excitation, 545 nm; emission, 590 nm) and H2O2 concentration was calculated from calibration curves generated by serial dilutions of standard H2O2. To evaluate the amount of H2O2 formed specifically via SSAO-mediated reaction, semicarbazide 100 μM was included in the control wells subjected to the same treatments and these values were subtracted from the total amount of H2O2 formed.
The inhibition was measured as % decrease of the signal compared to a control without inhibitor. Blank values in absence of substrates were subtracted from the fluorescence for each experimental condition. The IC50 shown in Table 1 were calculated with GraphPad Prism 4 program.
It should be understood that the foregoing disclosure emphasizes certain specific embodiments of the invention and that all modifications or alternatives equivalent thereto are within the spirit and scope of the invention as set forth in the appended claims.
The invention and the manner and process of making and using it, are now described in such full, clear, concise and exact terms as to enable any person skilled in the art to which it pertains, to make and use the same. It is to be understood that the foregoing describes preferred embodiments of the invention and that modifications may be made therein without departing from the spirit or scope of the invention as set forth in the claims. To particularly point out and distinctly claim the subject matter regarded as invention, the following claims conclude this specification.
This applications claims priority to U.S. provisional application Ser. No. 60/598,010, filed Aug. 2, 2004, the disclosure of which is explicitly incorporated by reference herein.
| Filing Document | Filing Date | Country | Kind | 371c Date |
|---|---|---|---|---|
| PCT/EP05/53778 | 8/2/2005 | WO | 00 | 5/9/2008 |
| Number | Date | Country | |
|---|---|---|---|
| 60598010 | Aug 2004 | US |
