Core 2glcnac-t inhibitors

Abstract

Treatments for conditions involving detrimental activity of the enzyme core 2 GlcNAc-T are provided using compounds of the formula I wherein R 1 is H, —OH, C 1-6 alkoxy, —NR 5 R 6 , or Sac 1; R 2 is H, —OH, C 1-6 alkoxy or Sac 2; R 3 is H, —OH, C 1-6 alkoxy or Sac 3; R 4 is H, C 1-6 alkyl, C 1-6 hydroxyalkyl or C 1-6 -alkoxy-C 1-6 -alkyl; R 5 is H, C 1-6 alkyl or C 1-6 acyl; R 6 is H, C 1-6 alkyl or C 1-6 acyl; Sac 1 Sac 2 and Sac 3 are independently selected saccharide moieties; and Z is a steroid moiety; or a pharmaceutically acceptable salt, ether or ester form thereof

1 . A method of treating a subject in need of therapy for a condition involving detrimental activity of the enzyme Core 2 GlcNAc-T comprising administration of a therapeutically effective amount of an isolated inhibitor of core 2 GlcNAc-T of formula IIa, IIb or IV to a patient in need thereof 
       
         
           
           
               
               
           
         
         wherein
 R 1  is —OH, C 1-6  alkoxy or —NR 5 R 6 ; 
 R 2  is —OH; 
 R 3  is —OH, 
 R 4  is H, C 1-6  alkyl, C 1-6  hydroxyalkyl; 
 R 5  is H, C 1-6  alkyl or C 1-6  acyl; 
 R 6  is H, C 1-6  alkyl or C 1-6  acyl; 
 Sac 1 Sac 2 and Sac 3 are independently selected saccharide moieties; and 
 Z is a steroid moiety 
 
         wherein the steroid moiety Z is of the formula VI: 
       
       
         
           
           
               
               
           
         
         which steroid moiety Z may incorporate a further group selected from the groups consisting of groups VI (a) to VI (e): 
       
       
         
           
           
               
               
           
         
         wherein:
 R 7  and R 14  are independently selected from H and —OH; 
 R 8  is C 1-6  alkyl; 
 R 9 , R 11 , R 16  are independently selected from H and C 1-6  alkyl; 
 R 10  is H or —OH or the H normally also present is absent and R 10  is ═O; 
 R 12  is H, —OH or C 1-6  acyl or a group selected from VII a or VII b; 
 
       
       
         
           
           
               
               
           
         
         
           R 13  is H. 
           R 15  is H, —OH or C 1-6  alkyl or R 13  and R 15  taken together form a —CH 2 —CH 2 -group; 
           R 17  is H, C 1-6  alkyl or C 1-6  hydroxyalkyl; 
           R 18 , R 23 , R 27  and R 33  are independently selected from C 1-6  alkyl; 
           R 19  and R 24  are independently selected from H and —OH. 
           R 20  is H, —OH or C 1-6  alkoxy or R 19  and R 20  taken together represent the second bond of a double bond joining adjacent carbon atoms; 
           R 21  is C 1-6  alkyl, C 2-6  alkenyl, C 2-6  alkynyl or a C 1-6  alkyl or C 2-6  alkenyl group substituted by one or more groups selected from the group consisting of —OH, C 1-6  alkoxy and Sac 4. 
           R 22  is H or —OH; 
           R 25  is C 1-6  alkyl, C 1-6  hydroxyalkyl or ═CH 2 . 
           R 26  is —OH; 
           R 28  is C 1-6  alkyl, C 2-6  alkenyl or C 2-6  alkynyl; 
           R 29  is C 1-6  alkyl; 
           R 30  is C 1-6  hydroxyalkyl; 
           R 31  is C 1-6  alkyl, C 1-6  hydroxyalkyl or C 1-6  alkyl substituted by Sac 5; 
           R 32  is C 1-8  alkyl, C 2-8  alkenyl or C 2-8  alkynyl; and 
           R 34  is C 1-6  hydroxyalkyl or C 1-6  alkyl substituted by Sac 6. 
           R 35  is C 1-6  alkyl; 
           Sac 4, Sac 5 and Sac 6 are independently selected monosaccharides; 
           X is either O or NH; and 
              represents a bond that is either double or single. 
         
         or a pharmaceutically acceptable salt, ether or ester form thereof; and 
         wherein said condition involving detrimental activity of the enzyme Core 2 GlcNAc-T is selected from the group consisting of inflammation, multiple sclerosis, myopathy, diabetic cardiomyopathy, myocardial dysfunction, retinopathy, diabetic retinopathy, nephropathy, atherosclerosis, asthma, rheumatoid arthritis, inflammatory bowel disease, transplant rejection, ischemia reperfusion injury, restenosis, ileitis, Crohn's disease, thrombosis, cholitis, lupus, frost bite injury, acute leukocyte mediated lung injury, traumatic shock, septic shock, nephritis, psoriasis, cholicytitis, cirrhosis, diverticulitis, fulminant hepatitis, gastritis, gastric and duodenal ulcers, hepatorenal syndrome, irritable bowel syndrome, jaundice, pancreatitis, ulcerative cholitis, human granulocyte ehlichiosis, T-cell activation, AIDS, infection with viruses, bacteria, protozoa and parasites adapted to use particular core 2 derived glycans, and cancer metastasis. 
       
     
     
         2 . A method according to  claim 1  wherein the steroid group Z is a steroid group of the formula VI, which steroid group of the formula VI incorporates a further group VIa wherein
 R 9  is H, 
 R 11  is C 1-6  alkyl; 
 R 12  is H or —OH; 
 R 16  and R 17  are H and 
 R 21  is a C 2-6  alkenyl group or a C 1-6  alkyl or C 2-6  alkenyl group substituted by one or more groups selected from the group consisting of —OH, —OCH 3  and Sac 4 
 
     
     
         3 . A method according to  claim 1  wherein the steroid group Z is a steroid group of the formula VI, which steroid group of the formula VI incorporates a further group VIc wherein
 R 8  is C 1-6  alkyl, 
 R 9  is H, 
 R 11  is C 1-6  alkyl; 
 R 12  is H or —OH; 
 R 15  is H or —OH; 
 R 16  and R 17  are H; 
 
     
     
         4 . A method according to  claim 1  wherein R 4  is CH 2 OH 
     
     
         5 . A method according to  claim 4  wherein the ring A is galactose or glucose. 
     
     
         6 . A method according to  claim 1  in which Sac 1 is rhamnose and Sac 2 and Sac 3 are glucose. 
     
     
         7 . The method according to  claim 2  in which the said isolated inhibitor of core 2 GlcNAc-T is selected from the group consisting of Trigoneoside IVa, glycoside F, Pardarinoside C, Pardarinoside D, Shatavarin I, Compounds 8a, 9a 10a and 25a of table 1a, protogracillin protoneogracillin methylprotogracillin, methylprotoneogracillin, pseudoprotogracillin, dracenoside Q dioscoreside E, dracenoside P tuberoside C and icogenin 
     
     
         8 . The method according to  claim 3  wherein the said isolated inhibitor of core 2 GlcNAc-T is selected from the group consisting of balanitin VI, deltonin, shatavarin IV and compounds 12a, 13a, 14a, 15a, 16a, 17a, 18a, 19a and 20a of table 1b. 
     
     
         9 . A method of treatment of multiple sclerosis in a subject comprising administering to the subject in need thereof, a therapeutically effective amount of a compound capable of reducing the activity of Core 2 GlcNAc-T. 
     
     
         10 . The method according to  claim 9  wherein the substance capable of reducing the activity of Core 2 GlcNAc-T is an inhibitor of Core 2 GlcNAc-T enzyme activity. 
     
     
         11 . The method according to  claim 10  wherein the compound is selected from the group consisting of steroidal glycosides, analogues of Uridine Diphosphate-N-Acetylglucosamine, analogues of Uridine Diphosphate, βGal(1→3)α(6-deoxy)GalNAcα-Bn and those compounds obtainable by UV activation of a compounds selected from the group consisting of Galβ1→3GalNAcα-pnp, Galβ1→3GalNAcα-onp, GalNAcα-pnp, GalNAcβ-pnp, GlcNAcβ-pnp, Galβ-pnp, GlcNAcβ1→3GalNAcα-pnp, L-Fucα1→2Galβ-pnp, GlcNAcα-pnp, Galβ1→6GlcNAcβ-pnp and Galβ1→3GlcNAcβ-pnp. 
     
     
         12 . The method according to  claim 9  wherein the compound capable of reducing the activity of Core 2 GlcNAc-T is an inhibitor of protein kinase C. 
     
     
         13 . The method according to  claim 12  wherein the compound is selected from the group consisting of 3,4-di-indoyl-pyrrol-2,5-dione derivatives. 
     
     
         14 . A method according to  claim 12  wherein the compound is ruboxystaurin. 
     
     
         15 . A method of according to  claim 9  wherein the compound is selected from the group consisting of Protodioscin, pseudoprotodioscin, protoneodioscin, methylprotodioscin, Trigoneoside IVa, Pardarinoside C, Pardarinoside D, dioscin, Balanitin VI, Deltonin, Shatavarin I, Shatavarin IV, protogracillin, proto neogracillin, methylprotogracillin, methylprotoneogracillin, pseudoprotogracillin, dracenoside Q, dioscoreside E, dracenoside P, tuberoside C, icogenin, gracillin, collettiside IV, 17-OH gracillin dracaenoside H, dracaenoside L, dracaenoside I, lilioglycoside H, lilioglycoside I, dracaenoside D, neoalsoside A, neoalsoside C, hoduloside V, Lotoside II and the following as yet unnamed compounds Compound 8a, 12a, 13a 14a 15a, 16a, 17a, 18a, 23a, 24a, 25a and 26a, of table 1a, 1b and 1c; and compounds 17b, 21b and 25b of table 3a 3b and 3c. 
     
     
         16 . A method of diagnosing Multiple Sclerosis in a subject comprising comparing the level of Core 2 GlcNAc-T activity associated with samples isolated from the subject with the level of Core 2 GlcNAc-T activity determined in samples isolated from healthy non afflicted individuals, a level of Core 2 GlcNAc-T activity higher than that in samples isolated from healthy non afflicted individuals being indicative that the subject is afflicted with MS. 
     
     
         17 . A method according to  claim 16  wherein a level of Core 2 GlcNAc-T activity associated with samples isolated from the subject that is least 2 times higher than that in samples isolated from healthy non afflicted individuals is indicative of MS in the subject. 
     
     
         18 . A method according to  claim 16  in which the samples isolated from the subject are biopsy samples or blood samples. 
     
     
         19 . A method according to  claim 16  in which the samples isolated from the subject are leukocytes. 
     
     
         20 . A method of determining the utility of a substance for use in the treatment of Multiple Sclerosis comprising measuring the ability of the substance to inhibit the activity of Core 2 GlcNAc-T. 
     
     
         21 . A food or beverage product incorporating an isolated Core 2 GlcNAc-T inhibitor. 
     
     
         22 . A method of treatment of conditions associated with raised activity of the enzyme core 2 GlcNAc-T comprising administration of an effective amount of a compound of the formula I to a patient in need thereof. 
       
         
           
           
               
               
           
         
         wherein R 101  is —OH, C 1-6  alkoxy, —NR 108 R 109 , or a monosaccharide of the formula: IXa: 
       
       
         
           
           
               
               
           
         
         R 102  is —OH, C 1-6  alkoxy or a monosaccharide of the formula IXb; R 103  is —OH, C 1-6  alkoxy or a monosaccharide of the formula IXc; R 104  is C 1-6  alkyl, C 1-6  hydroxyalkyl or C 1-6 -alkoxy-C 1-6 -alkyl; R 105  is C 1-6  alkyl, C 1-6  hydroxyalkyl or C 1-6 -alkoxy-C 1-6 -alkyl; R 106  is C 1-6  alkyl, C 1-6  hydroxyalkyl or C 1-6 -alkoxy-C 1-6 -alkyl; R 107  is C 2-6  alkyl, C 1-6  hydroxyalkyl or C 1-6 -alkoxy-C 1-6 -alkyl; R 108  is H, C 1-6  alkyl or C 1-6  acyl; R 109  is H, C 1-6  alkyl or C 1-6  acyl; and 
         Z is a steroid group selected from the group consisting of formula XIII formula XV and formula XVI; 
       
       
         
           
           
               
               
           
         
       
       wherein
 R 122  is H, OH, C 1-6  alkyl or C 1-6  alkoxy; R 113  is H, —OH, ═O, or C 1-6  alkyl; R 114  is H, —OH or C 1-6  alkyl or R 114  and R 133  taken together represent the second bond of a double bond joining adjacent carbon atoms; R 115  is H, or —OH, or R 115  and R1 33  taken together are ═O; R 116  is H, OH or ═O; R 117  is H, OH or ═O; R 118  is H, OH, C 1-6  alkoxy or C 1-6  alkyl; R 119  is H, OH, C 1-6  alkyl or C 1-6  alkoxy; R 120  is H, OH, C 1-6  alkoxy or C 1-6  alkyl; R 121  is H, OH, C 1-6  alkyl, C 1-6  alkoxy or is a group of the formula XIV: 
 
       
         
           
           
               
               
           
         
         R 122  is H, OH, C 1-6  alkyl or C 1-6  alkoxy; R 123  is H, OH, C 1-6  alkyl, C 1-6  hydroxyalkyl, C 1-6 -alkoxy-C 1-6 -alkyl, ═CH 2  or ═CH—C 1-6 -alkyl; R 124  is H, C 1-6  alkyl, C 1-6  acyl or a monosaccharide Sac 7; R 128  and R 129  are the same or different and are H or —OH; R 132  is H, OH or ═O; R 133  is H, or R 133  and R 115  taken together are ═O, or R 133  and R 114  taken together represent the second bond of a double bond joining adjacent carbon atoms; Sac 7 is selected from a group consisting of arabinose, xylose, lyxose, ribose, glucose, mannose, galactose, allose, altrose, gulose, idose, talose, ribulose, xylulose, fructose, sorbose, tagatose, psicose, sedoheptulose, deoxyribose, fucose, rhamnose, 2-deoxy-glucose, quinovose, abequose, glucosamine, mannosamine, galactosamine, neuraminic acid, muramic acid, N-acetyl-glucosamine, N-acetyl-mannosamine, N-acetyl-galactosamine, N-acetylneuraminic acid, N-acetylmuramic acid, O-acetylneuraminic acid, N-glycolylneuraminic acid, fructuronic acid, tagaturonic acid, glucuronic acid, mannuronic acid, galacturonic acid, iduronic acid, sialic acid and guluronic acid; preferably Sac 7 is selected from a group consisting of glucose, galactose, mannose, fucose, N-acetyl-glucosamine, N-acetyl-galactosamine and sialic acid; most preferably Sac 7 is glucose; and Y is N or O; 
       
       
         
           
           
               
               
           
         
         wherein: 
         R 112  is H, —OH, C 1-6  alkyl or C 1-6  alkoxy; R 113  is H, —OH, ═O, or C 1-6  alkyl; R 114  is H—OH or C 1-6  alkyl or R 114  and R 13   3  taken together represent the second bond of a double bond joining adjacent carbon atoms; R 115  is H, or —OH, or R 115  and R 133  taken together are ═O; R 116  is H, —OH or ═O; R 117  is H, —OH or ═O; R 118  is H, —OH, C 1-6  alkoxy or C 1-6  alkyl; R 119  is H, —OH, C 1-6  alkyl or C 1-6  alkoxy; R 120  is H, —OH, C 1-6  alkoxy or C 1-6  alkyl; R 127  is H, —OH, C 1-6  alkyl, C 1-6  alkoxy or C 1-6  hydroxyalkyl; R 128  and R 129  are the same or different and are H or —OH; R 132  is H, —OH or ═O; and R 133  is H, or R 133  and R 115  taken together are ═O, or R 133  and R 114  taken together represent the second bond of a double bond joining adjacent carbon atoms; 
       
       
         
           
           
               
               
           
         
         wherein: 
         R 112  is H, OH, C 1-6  alkyl or C 1-6  alkoxy; R 113  is H, —OH, ═O, or C 1-6  alkyl; R 114  is H, —OH or C 1-6  alkyl or R 114  and R1 33  taken together represent the second bond of a double bond joining adjacent carbon atoms; R 115  is H, or —OH, or R 115  and R 133  taken together are ═O; R 116  is H, —OH or ═O; R 117  is H, —OH or ═O; R 118  is H, —OH, C 1-6  alkoxy or C 1-6  alkyl; R 119  is H, —OH, C 1-6  alkyl or C 1-6  alkoxy; R 125  is H, —OH, C 1-6  alkyl or C 1-6  alkoxy; R 126  is H, —OH, C 1-6  alkyl, C 1-6  hydroxyalkyl, C 1-6 -alkoxy-C 1-6 -alkyl, ═CH 2  or ═CH—C 1-6 -alkyl; R 128  and R 129  are the same or different and are H or —OH; R 131  is H or —OH; R 132  is H, —OH or ═O; R 133  is H, or R 133  and R 115  taken together are ═O, or R 133  and R 114  taken together represent the second bond of a double bond joining adjacent carbon atoms; R 134  is H or —OH; and X is O, S or NH; 
       
       or a pharmaceutically acceptable salt, ester or tautomeric form or derivative thereof. 
     
     
         23 . A method according to  claim 22  wherein the compound of the formula VIII is selected from the group consisting of a compound of the formula X and a compound of the formula XII 
       
         
           
           
               
               
           
         
         wherein: 
         R 104  is C 1-6  alkyl, C 1-6  hydroxyalkyl or C 1-6 -alkoxy-C 1-6 -alkyl; R 105  is C 1-6  alkyl, C 1-6  hydroxyalkyl or C 1-6 -alkoxy-C 1-6 -alkyl; R 107  is C 2-6  alkyl, C 1-6  hydroxyalkyl or C 1-6 -alkoxy-C 1-6 -alkyl; 
       
       
         
           
           
               
               
           
         
       
       wherein
 R 101  is —OH, C 1-6  alkoxy or NR 108 R 109 , or a monosaccharide of the formula IXa: 
 
       
         
           
           
               
               
           
         
         R 104  is C 1-6  alkyl, C 1-6  hydroxyalkyl or C 1-6 -alkoxy-C 1-6 -alkyl; R 105  is C 1-6  alkyl, C 1-6  hydroxyalkyl or C 1-6 -alkoxy-C 1-6 -alkyl; R 106  is C 1-6  alkyl, C 1-6  hydroxyalkyl or C 1-6 -alkoxy-C 1-6 -alkyl; R 108  is H, C 1-6  alkyl or C 1-6  acyl; and R 109  is H, C 1-6  alkyl or C 1-6  acyl;