Steroidal Glycoside Compounds As Core 2 Glcnac-T Inhibitors

(19) TZZ_ZZ _T

(11) EP 1 909 802 B1

(12) EUROPEAN PATENT SPECIFICATION

(45) Date of publication and mention (51) Int Cl.: of the grant of the patent: A61K 31/704 (2006.01) A61K 31/7048 (2006.01) 21.05.2014 Bulletin 2014/21 A61K 31/706 (2006.01) A61P 1/04 (2006.01) A61P 1/06 (2006.01) A61P 1/16 (2006.01) (2006.01) (2006.01) (21) Application number: 06755733.0 A61P 3/10 A61P 7/02 A61P 9/10 (2006.01) A61P 11/00 (2006.01) A61P 11/06 (2006.01) A61P 13/12 (2006.01) (22) Date of filing: 06.07.2006 A61P 17/06 (2006.01) A61P 31/18 (2006.01) A61P 19/02 (2006.01)

(86) International application number: PCT/GB2006/002518

(87) International publication number: WO 2007/003957 (11.01.2007 Gazette 2007/02)

(54) Steroidal glycoside compounds as core 2 GlcNAc-T inhibitors Steroidale Glycosid-Verbindungen als core 2 GlcNAc-T-Hemmer Glycosides steroïdiens en tant qu’inhibiteurs du core 2 GlcNAc-T

(84) Designated Contracting States: WO-A-2004/062675 WO-A-2005/060977 AT BE BG CH CY CZ DE DK EE ES FI FR GB GR WO-A-2005/120535 WO-A2-01/32679 HU IE IS IT LI LT LU LV MC NL PL PT RO SE SI WO-A2-02/069980 CN-A- 1 415 625 SK TR US-A- 4 602 003 US-A- 5 104 856 US-A1- 2003 148 962 (30) Priority: 06.07.2005 GB 0513881 • DATABASE WPI 7 January 2004 (2004-01-07), (43) Date of publication of application: Derwent Publications Ltd., London, GB; Class 16.04.2008 Bulletin 2008/16 042,page 3, AN 2004-239758 XP002409228 HUANG H, LIU Z: "Medicine composition for (60) Divisional application: treating myocardial ischemia, angina pectoris 11163561.1 / 2 382 979 and cardiac infarction" & CN 1 465 344 A 11163563.7 / 2 382 980 (CHENGDU DIAO PHARM GROUP CO LTD) 7 11163564.5 / 2 382 981 January 2004 (2004-01-07) • DATABASE WPI 2 March 2005 (2005-03-02), (73) Proprietor: BTG International Limited Derwent Publications Ltd., London, GB; Class London EC4M 7RD (GB) 054,page 4, AN 2005-426073 XP002409229 WANG JINGANG: "Dioscin oral disintegration tablet and (72) Inventor: CHIBBER, Rakesh its preparing method." & CN 1 586 493 A London WC2R 2LS (GB) (KEXINBICHENG MEDICINE SCIENCE) 2 March 2005 (2005-03-02) (74) Representative: Kirkham, Nicholas Andrew et al Graham Watt & Co. LLP St. Botolph’s House 7-9 St. Botolph’s Road Sevenoaks, Kent TN13 3AJ (GB)

(56) References cited: EP-A1- 1 800 685 WO-A-02/03996

Note: Within nine months of the publication of the mention of the grant of the European patent in the European Patent Bulletin, any person may give notice to the European Patent Office of opposition to that patent, in accordance with the Implementing Regulations. Notice of opposition shall not be deemed to have been filed until the opposition fee has been paid. (Art. 99(1) European Patent Convention). EP 1 909 802 B1

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• DATABASE WPI 12 January 2005 (2005-01-12), • BIENVENU JEAN-GUY ET AL: "Recombinant Derwent Publications Ltd., London, GB; Class soluble P-selectin glycoprotein ligand-1-Ig 062,page 9, AN 2002-751531 XP002409230 ZHU reduces restenosis through inhibition of platelet- DAYUAN ET AL.: "Furost saponine analogue and neutrophil adhesion after double angioplasty in its separation process and use" & CN 1 184 229 swine", CIRCULATION, vol. 103, no. 8, 27 C (SHANGHAI INST OF PHARMACOLOGY) 12 February 2001 (2001-02-27), pages 1128-1134, January 2005 (2005-01-12) -& CN 1 184 229 C ISSN: 0009-7322 (SHANGHAI INST OF PHARMACOLOGY [CN]) 12 • BUERKE MICHAEL ET AL: "Sialyl Lewis-x- January 2005 (2005-01-12) containing oligosaccharide attenuates • DATABASE WPI 18 May 2005 (2005-05-18), myocardial reperfusion injury in cats", JOURNAL Derwent Publications Ltd., London, GB; Class OF CLINICAL INVESTIGATION, vol. 93, no. 3, 056,page 5, AN 2005-631469 XP002409231 HAN 1994, pages 1140-1148, ISSN: 0021-9738 J. ET AL.: "Medicine for regulating blood fat and • MYERS DANIEL ET AL: "New and effective treating cardiocerehral disease and preparing treatment of experimentally induced venous method" & CN 1 615 896 A (YUNNAN PROV thrombosis with anti-inflammatory rPSGL-Ig", MEDICINE INST) 18 May 2005 (2005-05-18) THROMBOSIS AND HAEMOSTASIS, vol. 87, no. • DATABASE WPI 26 January 2000 (2000-01-26), 3, March 2002 (2002-03), pages 374-382, ISSN: Derwent Publications Ltd., London, GB; Class 0340-6245 003,page 9, AN 2000-443110 XP002409232 LI • MIMAKI YOSHIHIRO ET AL: "Inhibitory effects of PINGYA ET AL.: "Process for extracting steroidal saponins on 12-O- ginsenoside Re, and use of medicine thereof" & tetradecanoylphorbol-13-acetate CN 1 242 374 A (XINLIHENG PHARMACEUTICAL (TPA)-enhanced 32P-incorporation into SCIEN) 26 January 2000 (2000-01-26) phospholipids of HeLa cells and proliferation of • DATABASE WPI 1 January 1997 (1997-01-01), human malignant tumor cells", BIOLOGICAL Derwent Publications Ltd., London, GB; Class AND PHARMACEUTICAL BULLETIN, vol. 18, no. 980,page 9, AN 1998-087548 XP002409233 3, 1995, pages 467-469, ISSN: 0918-6158 JUNPENG PENG ET AL.: "Anti-thrombosis glucoside medicine" & CN 1 138 984 A (RADIOMEDICINE INST MILITARY ME) 1 January 1997 (1997-01-01) • ZHANG, JIANYING ET AL: "Effect of six steroidal saponins isolated from Anemarrhenae rhizoma on platelet aggregation and hemolysis in human blood" CLINICA CHIMICA ACTA , 289 (1-2), 79-88 CODEN: CCATAR; ISSN: 0009-8981, 1999, XP002409220 • DATABASE WPI 6 April 2006 (2006-04-06), Derwent Publications Ltd., London, GB; Class 062,page 8, AN 2006-272298 XP002409234 FU T. ET AL.: "Steroidal saponin pharmaceutical composition, its preparation method and use." & WO 2006/034655 A (CHENGDU DIAO PHARM GROUP CO LTD) 6 April 2006 (2006-04-06) -& WO 2006/034655 A (CHENGDU DI AO PHARMACEUTICAL G [CN]; LIU ZHONGRONG [CN]; QI WEI [CN];) 6 April 2006 (2006-04-06)

2 EP 1 909 802 B1

Description

[0001] The present invention relates to the use of known and novel compounds as pharmaceutical actives against diseases susceptible to treatment by modulation, eg. inhibition, of the enzyme Core 2 GlcNAc- transferase (EC 2.4.1.102), 5 also known as UDP-GlcNAc:Galβ1,3GalNAc-R (GlcNAc to GalNAc) β-1,6-N-acetylglucosaminyl transferase (core 2 β- 1,6 N-acetylaminotransferase, hereinafter referred to as Core 2 GlcNAc-T, as defined in the claims. [0002] Inhibitors of Core 2 GlcNAc-T are known but none are in clinical development as isolated actives for pharmaceutical use. Examples of known compounds are disclosed in WO0187548, Kuhns et al., Glycoconjugate Journal 10 381-394 (1993), Hindsgaul et al., J Biol Chem. 266(27):17858-62 (1991), and Toki et al, Biochem Biophys Res Commun. 10 198(2):417-23 (1994). [0003] Applicant’s co-pending application WO05/060977 discloses known and novel steroidal glycosides that have therapeutic use as Core GlcNAc-T inhibitors, discusses the basis for use of such inhibitors in therapy and discloses published documents detailing the basis for Core 2 GlcNAc-T involvement in a number of diseases. The present application discloses further steroidal glycoside compounds that are inhibitors of core 2 GlcNAc-T and additional conditions 15 in which these compounds have a therapeutic use. [0004] Some of the presently disclosed steroidal glycosides have been tested previously in a limited number of disease paradigms. For example in protection against gastric mucosal lesions in rats (Matsuda H et al Bioorg Med Chem Lett. 24;13(6):1101-6, 2003), in mouse ear edema tests for anti inflammatory activity (Kim et al Arch Pharm Res. 22(3):313-6 (1999)), in treatment of dementia (US6593301) as "immuno-modulators" and spermatogenesis and ovulation stimulators 20 (Vasil’eva and Paseshnichenko Adv. Exp. Med. Biol. 404, 15-22 (1996)) and as adjuvants (Oda et al Biol Chem. 381(1):67-74 (2000)). Compounds of the invention have also been used in cytotoxicity assays (e.g. Hu et al Planta Medica, 63(2), 161-165 (1997), Mimaki et al Phytochemistry 37(1):227-32 (1994), Akhov et al Proc. Phytochem. Soc. Euprope 45, 227-231 (2000)), however cytotoxic concentrations are several orders of magnitude higher than those currently disclosed for inhibition of Core 2 GlcNAc-T activity. None of the aforementioned publications discloses that 25 certain steroidal glycosides are inhibitors of Core 2 GlcNAc-T. [0005] CN 1586493 A discloses a dioscin oral disintegration tablet ands its preparing method. CN 1415625 A discloses a method for preparing yam saponin, medical preparation and new usage in medication [0006] Certain plant sterol compounds, some of which are used as dietary supplements, impede the uptake of cholesterol from the gut and consequently lower plasma LDL cholesterol. However these compounds are generally used 30 in doses of several grams per day and are not known to be inhibitors of Core 2 GlcNAc-T. [0007] In a first aspect the present invention provides a compound of formula III as defined in the claims or a pharmaceutically acceptable salt, ether or ester thereof for use in the treatment of a subject in need of therapy for a condition involving detrimental activity of the enzyme core 2 GlcNAc-T, particularly raised activity, wherein the condition is selected from the group consisting of ischemia reperfusion injury, restenosis and thrombosis. 35 [0008] The prior art associates Core 2 GlcNAc-T (particularly through its involvement with branched oligosaccharide synthesis) with inter alia, vascular diseases, (including complications of diabetes), autoimmune and inflammatory conditions. Particular conditions associated with this enzyme and thus subject to treatment by the present invention are myopathy, retinopathy, nephropathy, atherosclerosis, asthma, rheumatoid arthritis, inflammatory bowel disease, trans- plant rejection, ischemia reperfusion injury (eg stroke, myocardial ischemia, intestinal reperfusion e.g. after hemorrhagic 40 shock,), restenosis, ileitis, Crohn’s disease, thrombosis, cholitis including for example ulcerative cholitis), lupus, frost bite injury, acute leukocyte mediated lung injury (eg adult respiratory distress syndrome), traumatic shock, septic shock, nephritis, psoriasis, cholicytitis, cirrhosis, diverticulitis, fulminant hepatitis, gastritis, gastric and duodenal ulcers, hepa- torenal syndrome, irritable bowel syndrome, jaundice, pancreatitis, ulcerative cholitis, human granulocyte ehlichiosis, Wiskott-Aldrich syndrome T-cell activation, AIDS, infection with viruses, bacteria, protozoa and parasites adapted to use 45 particular core 2 derived glycans and cancer. Cancer metastasis is particularly treatable by the present method. [0009] Cancers may include, for example, leukemias, lymphomas, melanomas, adenomas, sarcomas, and carcinomas of solid tissues; particularly cancers include prostate, testicular, mammary, pancreatic, cervical, uterine, kidney, lung, rectum, breast, gastric, thyroid, neck, cervix, bowel, salivary gland, bile duct, pelvis, mediastinum, urethra, bronchogenic, bladder, esophagus, colon, small intestine and sarcomas (eg. Kaposi’s sarcoma) and adenomatous polyps. Particularly 50 susceptible cancers for treatment are oral cavity carcinomas, pulmonary cancers such as pulmonary adenocarcinoma, colorectal cancer, bladder carcinoma, liver tumours, stomach tumours colon tumours, prostate cancer, testicular tumour, mammary cancer, lung tumours oral cavity carcinomas. Particular application is found in cancer or its metastasis where Core 2 GlcNAc-T activity is raised. [0010] The compounds according to the invention are compounds of formula III: 55

3 EP 1 909 802 B1

wherein Z is defined in the claims. 10 [0011] Wherein the ring A is a glucose moiety, and which formula may be written

[0012] In which Rha represents rhamnose, Glc represents glucose and 2 and 4 are the positions of ring A to which the saccharides are attached. 20 [0013] Most preferred are compounds which are 6-deoxy- α-L-mannopyranosyl-(1→2)-O-[β-D-glucopyranosyl-(1→4)]- β-D-glucopyranosides of steroid moieties Z. [0014] The term "steroid moiety" denotes a moiety comprising a tetracyclic ring system shown as formula V:

[0015] Typically the steroid moiety ring system is modified, for example by the addition of one or more further rings and/or one or more double bonds and/or one or more substituents. Typically the saccharide ring A is attached to the steroid moiety at the 3 position. The steroid moiety may for example have the ring system of cholestane, pregnane, 35 androstane, estrane, cholesterol, cholane, progestin, a mineralocorticoid, such as dehydroepiandrosterone or its 7-keto or 7-hydroxy analogue or a bile acid. [0016] In one disclosure the steroid moiety is that of a steroid that is in itself beneficial or neutral. By neutral is meant that the steroid ring is that which is considered suitable, whether as approved eg. by the FDA or as GRAS, for use in a human or animal. By beneficial is meant that the steroid has effects of benefit to the human or animal if it were administered 40 separately. [0017] The steroid moiety Z may for example be that of a steroidal sapogenin derivable from natural sources (for example plant sources) or a steroidal moiety which is itself derivable from such steroidal sapogenins by chemical mod- ification. The sapogenin may for example be that of a furostanol glycoside, a spirostanol glycoside (including those with nitrogen and oxygen containing rings) a damarane glycoside or other steroidal saponin. The steroid moiety Z according 45 to the invention is a steroid moiety of the formula VI

4 EP 1 909 802 B1

wherein the substituents have the meaning as indicated in the claims. [0018] Groups or rings that are incorporated into the steroid core VI are selected from those set out in formulae VI a and VI c wherein the dotted lines represent the relevant rings of the steroid core.

wherein:

20 R7 and R14 are independently selected from H and -OH; 8 8 R is C1-6 alkyl; preferably R is -CH3; 9 11 16 9 11 16 R , R and R are independently selected from H and C1-6 alkyl; preferably R , R and R are independently selected from H and -CH3; R10 is H or -OH or the H normally also present is absent and R 10 is =O; 25 12 12 R is H, -OH or C1-6 acyl; preferably R is H, -OH or acetyl; R13 is H. 15 13 15 15 R is H, C 1-6 alkyl or -OH or R and R taken together form a -CH 2-CH2- group; preferably R is H, -OH or -CH 3 13 15 or R and R taken together form a -CH2-CH2- group; 17 17 R is H, C1-6 alkyl or C1-6 hydroxyalkyl; preferably R is H, -CH2OH, or -CH3. 30 18 23 R and R are independently selected from C1-6 alkyl; R18 and R24 are independently selected from H and -OH; 20 19 20 R is H, -OH or C1-6 alkoxy or R and R taken together represent the second bond of a double bond joining 20 19 20 adjacent carbon atoms; preferably R is H, -OH or -OCH3 or R and R taken together represent the second bond of a double bond joining adjacent carbon atoms; 35 21 R is C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl or a C1-6 alkyl or C2-6 alkenyl group substituted by one or more groups 21 selected from the group consisting of -OH, C1-6 alkoxy and Sac 4; preferably R is C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl or a C1-6 alkyl group substituted by one or more groups selected from the group consisting of -OH, 1-6C 21 alkoxy and Sac 4; more preferably R is C2-6 alkenyl, or a C1-6 alkyl group substituted by one or more groups 21 selected from the group consisting of -OH, C 1-6 alkoxy and Sac 4; more preferably R is C2-6 alkenyl or a C 1-6 alkyl 40 group substituted by one or more groups selected from the group consisting of -OH, -OCH 3 and Sac 4; most preferably R21 is selected from the group consisting of 3-methyl but-2-eneyl, 2-methyl-prop-2-enyl, 3-methylbutanyl substituted at the 4-position by Sac4, 1-hydroxy-3-methylbutanyl substituted at the 4-position by Sac4, or 1-methoxy-3-methylbutanyl substituted at the 4-position by Sac4, R21 may be 3-methylenebutyl substituted at the 4-position by Sac4. 45 R21 may be 4-hydroxy-3-methylbutanyl. 25 25 R is C1-6 alkyl, C1-6 hydroxyalkyl or =CH2; preferably R is -CH3, -CH2OH or =CH2;

[0019] Sac 4 is a saccharide; preferably Sac 4 is a monosaccharide; more preferably Sac 4 is a hexose, a pentose or a tetrose; more preferably still Sac 4 is selected from glucose, galactose, quinovose, fucose, arabinose and xylose, 50 most preferably Sac 4 is glucose.

---- Represents a bond that is either double or single; and X is either O or NH; preferably X is O.

55 [0020] The steroid moieties Z are selected from groups VI which incorporate further groups VIa, or VIc. 9 11 12 [0021] Preferred steroid moieties Z incorporating further groups VI a are those in which R is H, R is C1-6 alkyl; R 16 17 21 is H or -OH; R and R are H and R is a C2-6 alkenyl group or a C1-6 alkyl or C2-6 alkenyl group substituted by one 21 or more groups selected from the group consisting of -OH, -OCH 3 and Sac 4, preferably R is a C2-6 alkenyl group or

5 EP 1 909 802 B1

a C1-6 alkyl group substituted by one or more groups selected from the group consisting of -OH, -OCH3 and Sac 4 8 9 11 12 16 17 21 preferably R is C1-6 alkyl and R is H and R is C1-6 alkyl; R is H or -OH; R and R are H and R is 3-methyl but- 2-eneyl, 2-methyl-prop-2-enyl, 3-methylbutanyl substituted at the 4-position by Sac4, 1-hydroxy-3-methylbutanyl substituted at the 4-position by Sac4, 3-methylenebutyl substituted at the 4-position by Sac 4 or 1-methoxy-3-methylbutanyl 5 substituted at the 4-position by Sac4; [0022] R21 may be 4-hydroxy-3-methylbutanyl 9 11 [0023] Alternatively, steroid moieties Z incorporating further groups VI a are those in which R is C1-6 alkyl; R is H; 20 21 9 11 20 21 10 R is H; and R is a C2-6 alkenyl; preferably R is C1-6 alkyl; R is H; R is H; R is a C2-6 alkenyl; and R is H; 9 11 20 21 10 15 more preferably R is C1-6 alkyl; R is H; R is H; R is a C2-6 alkenyl; R is H; and R is -OH or -CH2-CH2-; more 10 9 11 20 21 10 15 16 17 preferably R is C1-6 alkyl; R is H; R is H; R is a C2-6 alkenyl; R is H; R is -OH or -CH2-CH2-; and R and R is C1-6 alkyl. 8 9 [0024] Preferred steroid moieties Z incorporating further groups VI c are those in which R is C1-6 alkyl and R is H 11 12 15 16 17 and R is C 1-6 alkyl; R is H or -OH; R is H or -OH; R and R are H; more preferred steroid moieties Z incorporating 8 9 11 12 15 further groups VI c are those in which R is C1-6 alkyl and R is H and R is C1-6 alkyl; R is H or -OH; R is H or -OH; 15 R16 and R17 are H and X is O. [0025] Preferred steroid moieties of formula VI a are those having the ring structures illustrated in figure 1 still more preferably having the substitutions as set forth therein. In each case the chiral centre at the carbon labelled "25" can be in either the R or S configuration. [0026] More preferred steroid moieties, Z, of the formula VI c in which X = O are for example those having the radicals 20 of sarsasapogenin, smilagenin, 12β-hydroxysmilagenin, rhodeasapogenin, isorhodiasapogenin, samogenin, 12β-hydroxysamogenin, markogenin, yonogenin, convallagenin A, convallagenin B, tokorogenin, tigogenin, neotigogenin, gitogenin, agigenin digitogenin, chlorogenin, paniculogenin, (25R)-spirostan-3β, 17α,21-triol, allogenin, (25R)-5α- spirostan-2α,3β,5α,6α-tetraol, (24S,25R)-5α-spirostan-2α,3β,5α,6β,24-pentaol, yamogenin diosgenin, yuccagenin, lilagenin, ruscogenin, (25S)-ruscogenin, neopraserigenin, pennogenin, isonuatigenin, cepagenin, 24a-hydroxypennogen- 25 in, ophiogenin, sibiricogenin, convallamarogenin, neoruscogenin, hecogenin, neohecogenin, manogenin, sisalagenin and hispigenin. [0027] Preferred steroid moieties, Z, of the formula VI c in which X = NH are for example those that have the radicals of: solasodine, soladulcidine, tomatidine and 5-dehydrotomatidine. [0028] Preferred steroidal moieties Z of the formula VI c are those having the ring structures illustrated in figure 2; still 30 more preferably having the substitutions as set forth therein. [0029] Disclosed steroidal moieties Z of the formula VI are those having the ring structures of figure 3; still more preferably having the substitutions as set forth therein. [0030] Preferred disclosed steroid moieties VI i to VI xxxiii of figure 3 can be derived from steroidal glycoside compounds herein, of references of table 2 and additionally from the following references: Hostettman K. and Marston A (1995), 35 ibid., Mimaki et al Phytochemistry 37(1):227-32 (1994), Li et al Phytochemistry 29(12), 3893-8 (1990), Hernandez et al bioorganic and med. chem.. 12(16) 4423-4429 (2004), Renault et al, Phytochemistry, 44(7), 1321-1327 (1997). Zheng et al Steroids, 69(2), 111-119 (2004), Yoshikawa et al. Chemical & Pharmaceutical Bulletin, 40(9), 2287-91 (1992), Yoshikawa. et al Chemical & Pharmaceutical Bulletin, 40(9), 2275-8 (1992), Chen. et al Yunnan Zhiwu Yanjiu, 9(4), 495-502 (1987), Fujita et al Phytochemistry, 38(2), 465-72 (1995), Yin et al J. Nat. Products, 67(6), 942-952 (2004), 40 Sang Phytochemistry, 52(8), 1611-1615 (1999), Chen et al Yunnan Zhiwu Yanjiu, 6(1), 111-17 (1984). [0031] In one embodiment compounds of the invention are those of the formula III in which the steroid moiety -Z- is selected from group V which incorporate the further group VIa and in which R 7, R9, R10, R13, R14, R16, R17 and R19 are 12 8 11 18 15 20 21 H; R is H or -OH; R , R and R are -CH 3; R is H or -OH; R is -OH or -OCH 3 and R is 4-hydroxy-3-methylbutanyl, 3-methylenebutyl substituted at the 4-position by Glc, 3-methylbutanyl substituted at the 4-position by Glc, 1-hydroxy- 45 3-methylbutanyl substituted at the 4-position by Glc or 1-methoxy-3-methylbutanyl substituted at the 4-position by Glc. It is particularly preferred that when R 21 is 3-methylenebutyl substituted at the 4-position by Glc then the compound of the formula III is compound 25 of table 1a. and when R 21 is 4-hydroxy-3-methylbutanyl, then the compound of the formula III is either compound 9 or compound 10 of table 1. [0032] Particularly preferred compounds of the formula III are: 50 Trigoneoside IVa, glycoside F, Pardarinoside C, Pardarinoside D, Balanitin VI, Deltonin, Shatavarin I and Shatavarin IV.

[0033] Further preferred compounds that are so far un named are Compounds 8, 12, 13, 14, 15, 16, 17, 18, 23, 25 55 and 26 of table 2. [0034] The preferred compounds have the following chemical names:

Trigoneoside IVa is β,25S)-26-( (3 β-D-glucopyranosyloxy)-22-hydroxyfurost-5-en-3-yl O-6-deoxy-α-L-man-

6 EP 1 909 802 B1

nopyranosyl-(1→2)-O-[β-D-glucopyranosyl-(1→4)]-β-D-glucopyranoside, glycoside F is (3β,25R)-26-(β-D-glucopyranosyloxy)-22-hydroxyfurost-5-en-3-yl O-6-deoxy-α-L-mannopyranosyl-(1→2)-O-[β-D-glucopyranosyl-(1→4)]-β- D-glucopyranoside, Pardarinoside C is (3β,5α,22α,25R)-26-(acetyloxy)-14,17-dihydroxy-22-methoxyfurostan-3-yl O-6-deoxy-α-L-annopyranosyl-(1→2)-O-[β-D-glucopyranosyl-(1→4)]-β-D-Glucopyranoside, Pardarinoside D is β- 5 D-glucopyranoside, (3β,5α,22α,25R)-26-(acetyloxy)-17-hydroxy-22-methoxyfurostan-3-yl O-6-deoxy-α-L-mannopyranosyl-(1→2)-O-[β-D-glucopyranosyl-(1→4)]-β-D-glucopyranoside, Balanitin VI is (3β,25S)-spirost-5-en-3- yl-O-α-L-rhamnopyranosyl-(1→2)-O-[β-D-glucopyranosyl-β-D-glucopyranoside, Deltonin is (3β,25R)-spirost-5-en- 3-yl-O-α-L-rhamnopyranosyl-(1→2)-O-[β-D-glucopyranosyl-β-D-glucopyranoside, Shatavarin I is (3 β,5β,22α,25S)- 26-(β-D-glucopyranosyloxy)-22-hydroxyfurostan-3-yl O-6-deoxy-α-L-mannopyranosyl-(1→2)-O-[β-D-glucopyrano- 10 syl-(1→4)]-β-D-glucopyranoside, Shatavarin IV is β (3,5β,25S)-spirostan-3-yl O-6-deoxy-α-L-mannopyranosyl-(1→2)-O-[β-D-glucopyranosyl-β-D-glucopyranoside, Compound 8 is (3β,25R)-26-(β-D-glucopyranosyloxy)-22-methoxyfurost-5-en-3-yl O-6-deoxy-α-L-mannopyranosyl-(1→2)-O-[β-D-glucopyranosyl-(1→4)]-β-D-Gluco-pyranoside, compound 12 is β [(3,12α,25R)-12-hydroxyspirostan-3-yl O-6-deoxy-α-L-mannopyranosyl-(1→2)--[β-D-glucopyranosyl-(1→4)]-β-D-glucopyranoside], com- 15 pound 13 is [(25S)-spirost-5-ene-3β27-diol 3-O-{6-deoxy-α-L-mannopyranosyl-(1→2)--[β-D-glucopyranosyl-(1→4)]-β-D-glucopyranoside}], compound 14 is [(25R, 26R)-26-methoxyspirost-5-en-3β-ol 3-O-{6-deoxy-α-L- mannopyranosyl-(1→2)-[β-D-glucopyranosyl-(1→4)]-β-D-glucopyranoside}], compound 15 is β [3,25R,27(S)]- 27-(4-carboxy-3-hydroxy-3-methyl-1-oxobutoxy)spirost-5-en-3-yl O-6-deoxy-α-L-mannopyranosyl-(1→2)-O-[β-D- glucopyranosyl-(1→4)]-β-D-glucopyranoside], compound 16 is [3β,25R,27(S)]-27-[(3-hydroxy-5-methoxy-3-me- 20 thyl-1,5-dioxopentyl)oxy]spirost-5-en-3-yl O-6-deoxy-α-L-mannopyranosyl-(1→2)-O-[β-D-glucopyranosyl-(1→4)]- β-D-glucopyranoside], compound 17 is β-D-Glucopyranoside, (3β,25R,26R)-17-hydroxy-26-methoxyspirost-5-en- 3-yl O-6-deoxy-α-L-mannopyranosyl-(1→2)-O-[β-D-glucopyranosyl-(1→4)]-β-D-glucopyranoside, compound 18 is (3β,25R,26R)-26-hydroxyspirost-5-en-3-yl O-6-deoxy-α-L-mannopyranosyl-(1→2)-O-[β-D-glucopyranosyl-(1→4)]- β-D-glucopyranoside, compound 23 is 26-O-β-D-gluco-pyranosylnuatigenin 3-O-α-L-rhamnopyranosyl- 25 (1→2)-O-[β-D-glucopyranosyl-(1→4)]-β-D-glucopyranoside compound 25 is β (3,25S)-26-(β-D-gluco-pyranosyloxy)-22-hydroxyfurost-5, 25 (27) dien-3-yl O-6-deoxy-α-L-mannopyranosyl-(1→2)-O-[β-D-glucopyranosyl-(1→4)]- β-D-glucopyranoside, and compound 26 is solasodine 3-O-α-L-rhamnopyranosyl-(1→2)-O-[β-D-glucopyranosyl-(1→4)]-[β-D-glucopyranoside

30 [0035] Where any preferred substituent (such as C1-6 alkyl, C1-6 hydroxyalkyl) is said to be composed of from 1 to 6 carbon atoms (ie C1-6) such substituents are more preferred with 1 to 4 carbon atoms (ie C 1-4), are more preferred still with 1 or 2 carbon atoms (ie methyl or ethyl) and are most preferred with only one carbon atom (ie are in the methyl form). Likewise where partial substituents such as the C1-6 alkyl group or C1-6 alkoxy group of C1-6-alkoxy-C1-6-alkyl are said to be composed of from 1 to 6 carbon atoms (ie C1-6) such substituents are, independently one of the other, 35 more preferred with 1 to 4 carbon atoms (ie C1-4), are more preferred still with 1 or 2 carbon atoms (ie methyl or ethyl) and are most preferred with only one carbon atom (ie are in the methyl form). [0036] Alkyl, alkenyl and alykynyl radicals may, where the number of carbons in the chain permits, be either straight- chain or branched chain. C1-6 alkyl radicals may be, for example, methyl, ethyl, n-propyl or isopropyl, n-butyl, isobutyl or tert-butyl, isopentyl, 2,2-dimethyl propyl, n-hexyl, isohexyl and 1,3-dimethylbutyl. C2-6 alkenyl radicals may be, for 40 example, allyl, 1-methylprop-2-enyl, 2-methylprop-2-enyl, 2-methyl prop-1-enyl, but-2-enyl, but-3-enyl, 1-methyl-but-3- enyl, 1-methyl-but-2-enyl, 3-methylbut-2-enyl; where the alkenyl radical consists of 2-8 carbon atoms, the possible arrangements include, in addition to those possible for radicals with 2-6 carbon atoms, the following preferred radicals

5-methyl-hex-5-enyl, 4-methyl-hex-5-enyl, 3,4-dimethyl-hex-2-enyl. C 2-6. alkynyl may be, for example, propargyl, but-2- ynyl, but-3-ynyl, 1-methylbut-3-ynyl, pent-2-ynyl, pent-3-ynyl, pent-4-ynyl, 4-methyl-pent-2-ynyl. Preferably it is propargyl, 45 1-methylbut-3-ynyl, pent-2-ynyl, pent-4-ynyl or 4-methyl-pent-2-ynyl. [0037] A C1-6 hydroxyalkyl group may, where chemically possible, be either a C1-6 monohydroxyalkyl or a C1-6 dihy- droxyalkyl group. [0038] Where moieties may be, in turn, substituted by a saccharide moiety it is preferred that the bond is through an oxygen to form a group such as: 50

7 EP 1 909 802 B1

[0039] In the formula III the saccharide moiety A comprises multiple chiral centres. Thus each of the carbon atoms 1, 2, 3, 4 and 5 can, independently, be in the R or S form. Depending on the form of the anomeric carbon, A can, independently, be in either the alpha or beta anomeric form. For ring A the beta anomeric form is preferred. The saccharide moiety A can be in the D or L form; the D form is preferred. 5 [0040] Saccharides include monosaccharides, disaccharides, trisaccharides, tetrasaccharides and polysaccharides. Saccharide moieties may be monosaccharides, but may be independently selected as di- or oligosaccharides. [0041] Monosaccharides include tetroses pentoses, hexoses and heptoses, may be tetroses pentoses and hexoses. [0042] Tetroses may be for example aldotetroses, such as erithrose and threose and aldoketoses erithrulose. [0043] Pentosesinclude aldopentoses, such asarabinose, lyxose, riboseand xyloseand ketopentoses such as ribulose 10 and xylulose and deoxypentoses such as 2-deoxyribose and 3-deoxyribose. Pentoses may be xylose and arabinose. Pentoses may be in the furanose (eg arabinofuranose, lyxofuranose, ribofuranose and xylofuranose) or the pyranose (eg arabinopyranose, lyxopyranose, ribopyranose and xylopyranose) forms. [0044] Hexoses include aldohexoses, such as, allose, altrose, galactose, talose, gulose, idose, mannose and glucose (preferred are glucose, mannose, galactose, altrose, allose idose and talose) and ketokexoses such as fructose, psicose, 15 sorbose and tagatose. [0045] Hexoses may also be deoxy hexoses wherein an -OH group is replaced by an -H group at any position other than the bonded group. 6-deoxyhexoses are for example 6-deoxyallose, 6-deoxyaltrose, 6-deoxygalactose (fucose), 6- deoxyglucose (quinovose), 6-deoxygulose, 6-deoxyidose, 6-deoxymannose (rhamnose) or 6-deoxytalose. Deoxyhex- oses may also be 2-deoxy, 3-deoxy, 4-deoxy and 5-deoxy hexoses. The oxygen may be lacking at more than one 20 position. Examples of deoxyhexoses are - 2-deoxy-glucose, 2-deoxygalactose, 4-deoxyfucose, 3-deoxygalactose, 2- deoxyglucose, 3-deoxyglucose, 4-deoxyglucose. Deoxy-aldohexoses are preferred. [0046] Hexoses also include hexosamines such as galactosamine, glucosamine and mannosamine, n-acteyl hexosamines such as N-acetyl-galactosamine, N-acetyl-mannosamine and N-acetylglucosamine. Hexoses may be aldohexoses and deoxy hexoses, particularly glucose, galactose, quinovose, fucose and rhamnose. 25 [0047] Hexoses may be in the furanose or pyranose form; preferably in the pyranose form. [0048] Other monosaccharides include uronic acids, for example fructuronic acid, galacturonic acid, iduronic acid, glucuronic acid, guluronic acid,. mannuronic acid and tagat-uronic acid; sedoheptulose, sialic acid, neuraminic acid, muramic acid, N-acetylneuraminic acid, N-acetylmuramic acid, O-acetylneuraminic acid, and N-glycolylneuraminic acid. [0049] Hexoses may be aldohexoses and deoxyhexoses (particularly deoxyaldohexoses); pentoses may be aldopen- 30 toses and deoxy-pentoses (particularly deoxyaldopentoses). [0050] Pharmaceutically acceptable esters of compounds of the formula 1 are for example, an ester with an aliphatic or aromatic carboxylic or sulphonic acid. Aliphatic carboxylic acids may be for example of up to 6 carbon atoms, for example a methyl, ethyl, tert-butyl succinyl or malyl. Aromatic carboxylic acids may for example benzoic acid, sulphonic acids may be methylsulphonic or p-toluenesulphonic acid, and include esters at any available esterifiable position. 35 [0051] Pharmaceutically acceptable esters further include known compounds in which the sugar -OH groups are esterified with an aliphatic carboxylic acid of up to 6 carbon atoms. Also included are known esters at the carbon 26- position with compounds such as hydroxymethylgluteryric acid or its methyl ester (for example compound 19 and structure VI xxiv). [0052] Pharmaceutically acceptable ethers are, for example, with C 1-6 hydroxyalkyl compounds which may be formed 40 at any of the available -OH groups, for example on the saccharide moieties, or steroid moieties by converting one or more of the -OH groups to alkoxy groups (e.g. Li et al., Carbohydr Res. 20; 338(2): 117-21 (2003), Purdie and Irvine, J. Chem. Soc. 87, 1022 (1905), Haworth and Hirst, J. Chem. Soc. 119, 193 (1921). [0053] A suitable pharmaceutically-acceptable salt form of the compounds of the formula III is, for example, an acid addition salt with an inorganic or organic acid, for example hydrochloric, hydrobromic, trifluoroacetic or maleic acid; or 45 an alkali metal, for example sodium, an alkaline earth metal, for example calcium, or ammonium, for example tetra(2- hydroxyethyl)ammonium, salt. [0054] Compounds of the formula I can be extracted from a variety of plant species. Examples of sources of compounds of the invention and example purification protocols are given in the references of table 2. Further sources of compounds of the invention and methods of isolation of such compounds are detailed in Hostettman K. and Marston A. Saponins. 50 Cambridge University Press UK. (1995), particularly tables 2.2, 2.9, 2.10 and 2.11 and appendix 3 - and references therein. [0055] Many compounds of the invention are hydroxylated steroids. It is known in the art that such compounds, when exposed to solvent such as alcohols during purification or preparation, may be converted to alkoxy derivatives or to other derivatives such as methylketals (which revert to the original compounds upon drying). Particularly furostanol compounds of the formula VIa, in which the carbon at the at the 22-position of the furostanol structure, is substituted by -OH, may 55 be converted to alkoxy derivatives when exposed to alcohols. Notably such compounds may become methoxy derivatives when purified from plant sources using methanol-containing solvents. Alternatively they may be converted to the corre- sponding alkoxy by reflux in an appropriate anhydrous alcohol at elevated temperature, for example methanol (Hu. et al Planta Medica, 63(2), 161-165 (1997)). Such alkoxylated compounds are also compounds of the invention.

8 EP 1 909 802 B1

[0056] Where the compounds of the invention are purified from natural sources it is preferred that they are used in isolated form. By isolated is meant that the compound is at least 1% pure, conveniently it is at least 10% pure, more conveniently at least 30% pure, preferably it is at least 50% pure more preferably it is at least 80% pure still more preferably it is at least 90% pure and most preferably it is at least 95% pure. 5 [0057] The purity of the compound is conveniently expressed as a ratio of UV absorption associated with the compound to UV absorption associated with other material in the sample, conveniently at 205nm. The purity of the compound may be measured for example using a chromatography system such as for example TLC or HPLC such as are described in the references herein, particularly in those references relating to the compound in question, or in applicants co pending application WO05/060977 10 [0058] Alternatively, compounds of the invention can be synthesised via a number of routes known to the skilled worker. For example by glycosylation of appropriate aglycones. A number of suitable aglycones are available commer- cially, alternatively an suitable aglycone may be prepared, either by isolation from a natural source (27 and references therein), by deglycosylation of a suitable glycosylated compound (for example those compounds disclosed in Hostettman K. and Marston A. Saponins. Cambridge University Press UK. (1995) or herein), or by chemical synthesis from a variety 15 of starting material that are readily available. [0059] Methods of synthesising Gal β1-3(6-deoxy)GalNAcα-conjugates are given in Paulsen et al., Leibigs Ann. Chem. 747-758.(1992). These methods may be adapted by the skilled worker in combination with other methods referenced herein to synthesize other compounds of the formula III. [0060] The skilled worker will be aware of many sources of spirostanol and furostanol aglycones suitable for preparing 20 compounds for use in the invention. For example spirostanol aglycones wherein X=O or X=NH may be, for example, sarsapogenin, smilagenin, 12β-hydroxysmilagenin, Rhodeasapogenin, Isorhodiasapogenin, Samogenin, 12β-hydroxysamogenin, Markogenin, Yonogenin, Convallagenin A, Convallagenin B, Tokorogenin, Tigogenin, Neotigogenin, Gi- togenin, Agigenin Digitogenin, Chlorogenin, Paniculogenin, (25R)-Spirostan-3β, 17α21-triol, Allogenin, (25R)-5α- Spirostan-2α,3β,5α,6α-tetraol, (24S, 25R)-5α-Spirostan-2α,3β,5α,6β,24-pentaol, Yamogenin Diosgenin, Yuccagenin, 25 Lilagenin, Ruscogenin, (25S)-Ruscogenin, Neopraserigenin, Pennogenin, Isonuatigenin, Cepagenin, 24a-hydroxypennogenin, Ophiogenin, Sibiricogenin, Convallamarogenin, Neoruscogenin, Hecogenin, Neohecogenin, Manogenin, Si- salagenin, Solasodine, Soladulcidine, Tomatidine and 5-dehydrotomatidine. [0061] Deglycosylation of, for example steroidal glycosides, may be simply carried out by acid hydrolysis, for example in a 50:50 mix of 2N HCl: dioxane at 100°C in a sealed tube for 4.5 hrs (Hu K (1997) ibid). 30 [0062] Methods for the synthesis of a number of steroidal aglycones have been known for may years. For example synthesis of diosgenin, yamogenin, kryptogenin and isonarthogenin have been reported by Kessar et al., Tetrahedron; 24(2):905-7 (1968), Kessar et al., Tetrahedron 24(2):899-904 (1968) and Kessar et al., Tetrahedron. 24(2):887-92 (1968). [0063] General synthetic routes to a variety of tri saccharide substituted spirostanol saponins are known (Deng et al., Carbohydr Res.30;317(1-4):53-62. (1999), Li et al., Carbohydr Res.; 9;331(1):1-7. (2001), Yu et al., Tetrahedron letters, 35 42, 77-79 (2001). [0064] Yu et al., J Org Chem.; 13;67(25):9099-102 (2002)). Methods of synthesis of spirostanol saponins having 2,4 branched oligosaccharide moieties are also known (Cheng et al., J Org Chem. 2;68(9):3658-62 (2003), Deng et al 1999 ibid, Du et al., Org Lett.; 2;5(20):3627-30.(2003), Lehmann. et al,. Carbohydr Res. 337(21-23): 2153-9 (2002)). Methods of synthesis of furostanol saponins, synthesis of derivatised saponins and interconversion of spirostanol and furostanol 40 saponins have also been devised (Yu et al., J Comb Chem.; 3(5):404-6. (2001), Yu et al., J Org Chem.; 13;67(25):9099-102 (2002), Cheng et al., J Org Chem.; 2;68(9):3658-62 (2003), Du et al., Org Lett.; 2;5(20):3627-30.(2003), Li et al., Carbohydr Res. 20; 338(2): 117-21 (2003), Lehmann et al,. Carbohydr Res. 337(21-23): 2153-9 (2002), Tobari et al., Eur J Med Chem. 35(5): 511-27 (2000), Wang et al., Steroids. 69(10): 599-604 (2004)). [0065] Furthermore, furostanol and spirostanol saponins can be inter converted using a β-glucosidase (Inoue et al., 45 Phytochemistry 41(3), 725-7(1996)) and pseudosaponins maybe cyclised to form the spirostanol derivative (Tobari et al., Eur J Med Chem. 35(5): 511-27 (2000)). [0066] Combinatorial approaches to saponin synthesis have also been reported (Lautrette S. et al., Chem Commun (Camb). 7;(5): 586-7 (2004), Yu et al., J Comb Chem.; 3(5):404-6. (2001)). These references also provide information and further references on derivatisation of saccharide hydroxyl and hydroxyalkyl groups. 50 [0067] As used herein the term aglycone refers to steroidal glycosides wherein the saccharide moieties are not present. The compounds may have other substituents at the position originally occupied by the saccharide moiety. Particularly aglycones that are furostanol saponins when not glycosylated may be in the ring closed state as the equivalent spirostanol compounds. Steroidal glycosides are compounds having a steroid or substituted steroid core, to which is attached one or more saccharide moieties. A steroidal sapogenin is the aglycone of a steroidal saponin. A steroidal saponin is a 55 naturally derived steroidal glycoside. [0068] An anti cell adhesion agent is an agent that reduces the adhesion of cells to a substrate such as platelets or the lining of blood vessels or other tissues, an anti cell-cell interaction agent is an agent that reduces the interaction between cells. An anti cellular extravasation agent is an agent that reduces the passage of cells from the blood stream

9 EP 1 909 802 B1

through the walls of blood vessels. [0069] The term "treating", as used herein, includes treating as prophylaxis or treatment of a current or remitting illness. For the avoidance of doubt the term C 1-6 acyl is -CO-C1-5-alkyl. [0070] The use of compounds of the formula I I I in the manufacture of a medicament for the treatment of a condition 5 associated with detrimental activity, particularly raised activity, of the enzyme core 2 GlcNAc-T is disclosed [0071] The use of a compound of formula III as an anti cell adhesion agent, an anti cell-cell interaction agent or an anti cellular extravasation agent is further disclosed. [0072] A pharmaceutical composition comprising compound of the formula III is also disclosed. [0073] Medicaments of the invention comprising compounds of the formula I will typically be prepared in a sterile and 10 pyrogen free form. They can be administered by oral or parenteral routes, including intravenous, intramuscular, intraperitoneal, subcutaneous, transdermal, airway (aerosol), rectal, vaginal and topical (including buccal and sublingual) administration. [0074] The medicament may be made up in liquid form in which case it will typically, in addition to the compound of the formula III, comprise a pharmaceutically acceptable diluent or it may be made up in solid form. For oral administration, 15 the compounds of the invention will generally be provided in the form of tablets or capsules, as a powder or granules, or as an aqueous solution or suspension. Tablets for oral use may include the active ingredients mixed with pharmaceutically acceptable excipients such as inert diluents, disintegrating agents, binding agents, lubricating agents, sweet- ening agents, flavouring agents, colouring agents and preservatives. Examples of suitable inert diluents include sodium and calcium carbonate, sodium and calcium phosphate, and lactose, while corn starch and alginic acid are examples 20 of suitable disintegrating agents. Binding agents include, for example starch and gelatine, while the lubricating agent, if present, may for example, be magnesium stearate, stearic acid or talc. If desired, the tablets may be coated with an enteric coating material, such as glyceryl mono stearate or glyceryl distearate, to delay absorption in the gastrointestinal tract. Capsules for oral use include hard gelatine capsules in which the active ingredient is mixed with a solid diluent, and soft gelatine capsules wherein the active ingredients is mixed with water or an oil such as peanut oil, liquid paraffin 25 or olive oil [0075] Formulations for rectal administration may for example be presented as a suppository with a suitable base comprising, for example, cocoa butter or a salicylate. [0076] Formulations suitable for vaginal administration may for example be presented as pessaries, tampons, creams, gels, pastes, foams or spray formulations containing in addition to the active ingredient such carriers as are known in 30 the art to be appropriate. [0077] In preparations for intramuscular, intraperitoneal, subcutaneous and intravenous use, the compounds of the invention will typically be provided in a pharmaceutically acceptable diluent to provide sterile solutions, emulsions, liposome formulations or suspensions. Typically the preparation will be buffered to an appropriate pH and isotonicity. For example suitable diluents include Ringer’s solution and isotonic sodium chloride. Aqueous suspensions according 35 to the invention may include suspending agents such as cellulose derivatives, sodium alginate, polyvinylpyrrolidone and gum tragacanth, and a wetting agent such as lecithin. Suitable preservatives include ethyl and n-propyl p-hydroxyben- zoate. [0078] The isolated Core 2 GlcNAc-T inhibitors of the invention may also be incorporated into a food or beverage product. 40 [0079] In general a suitable dose of Core 2 GlcNAc-T inhibitor will be in the range of 10 ng to 50 mg per kilogram body weight of the recipient per day, preferably in the range 100 ng to 10 mg, more preferably in the range ofm g1 to 5.0 mg/kg/d. The desired dose is preferably presented once daily or several times a day in sub doses. These sub-doses may be administered in unit dosage forms, for example, containing 1mg to 1500 mg, preferably 40mg to 1000mg, and most preferably 50mg to 700 mg of active ingredient per unit dosage form. 45 [0080] In the shorthand annotation:

used in structures herein Glc is glucose and Rha is rhamnose. The annotation 2,3 and 2,4 denote the position of 55 attachment of the saccharides to the central monosaccharide. [0081] The shorthand notation

10 EP 1 909 802 B1

5 used in structures herein denotes the structure:

15 [0082] The present invention will now be described further by reference to the following Examples, Schemes and Figures.

FIGURES 20 [0083]

Figure 1 illustrates preferred steroid moieties Z of formula VIa. Figure 2 illustrates preferred steroidal moieties Z of the formula VI c. 25 Figure 3 illustrates preferred steroidal moieties Z of the formula VI (reference)

EXAMPLES

Table 1 Example compounds of the invention. 30 [0084]

45 Table 1a Compound Q A1 A2 A3 A4 A5 A6 A7 25 R/S Bond a Bond b 6 4-Glc Me H H Glc OH H H S Double Single 50 7 4-Glc Me H H Glc OH H H R Double Single 8 4-Glc Me H H Glc OMe H H R Double Single

94-GlcMeHOH-O.CO.CH3 OMe OH H R Single Single 10 4-Glc Me H OH -O.CO.CH OMe H H R Single Single 55 3 21 4-Glc Me H H Glc OH H H s Single Single

25 4-Glc =CH2 H H Glc OH H H - Double Single

11 EP 1 909 802 B1

(continued)

Compound Q A1 A2 A3 A4 A5 A6 A7 25 R/S Bond a Bond b 27*** 3-Glc Me H H Glc OH H H R Double Single 5 *** comparative compound

Table 1b

Comp. Q A1 A2 A3 A4 A5 A6 25R/S Bond a

25 12 4-Glc Me H H H OH H R Single

13 4-Glc -CH2OHHHHH H S Double 14 4-Glc Me H H H H OMe R Double 15 4-Glc * H H H H H R Double 30 16 4-Glc ** H H H H H R Double 17 4-Glc Me H OH H H OMe R Double 18 4-Glc Me H H H H OH R Double

35 19 4-Glc Me H H H H H S Double 20 4-Glc Me H H H H H R Double 22 4-Glc Me H H H H H S Single

12 EP 1 909 802 B1

Table 1c Further example compounds of the invention Compound Structure 23 5

Table 2: Key to example compounds of the invention and references 25 Compound Example references Compound name 6 3 Trigoneoside IVa 7 4, 3 Glycoside F, protodeltonin, deltoside, 30 85 No name 9 6 Pardarinoside C 10 6 Pardarinoside D 12 8 Not named 35 13 11, 9, 10, 5 Not named 14 9, 5 Not named 15 9, 10, 11, 12 Not named 40 16 10, 11 Not named 17 5 Not named 18 13 Not named

45 19 15, 5, 14 Balanitin VI 20 16, 17 Deltonin 21 19, 18 Shatavarin I 22 18, 23 Shatavarin IV 50 23 11 Not named 25 WO05/060977 Not named 26 11 Not named

55 27 1 Protogracillin *** *** comparative example compound.

13 EP 1 909 802 B1

Example 1. Biological activity of compounds.

[0085] All compounds used herein were supplied by Chromadex Inc. 2952 S. Daimler Street Santa Ana CA 92705. Compounds used were at least 88% pure 5 Table 4: Purity of compounds used Compound Number Purity Trigoneoside IVa 6 89% 10 Glycoside F 7 80.3% Shatavarin I 21 >95% ***Protogracillin 26 98.8% Purity was determined by HPLC using UV 15 absorption at 205nm

1a. Cell culture

20 [0086] The human leukocytic cell-line (U937) was cultured in RPMI supplemented with 10% foetal calf serum, 2 mM glutamine, 100 IU/ml penicillin and 100 mg/ml streptomycin.

1b. Assay of core 2 GlcNAc-T activity

25 [0087]

(i). Glucose induction of Core 2 GlcNAc-T leukocytes (U937 cells) were exposed to normal glucose (5.8 mM) or high glucose (15 mM) for 24 hours at 37°C. After incubation, the cells maybe lysed and frozen at -20°C until used for the measurement of core 2 GlcNAc-T. or used immediately. 30 (ii). TNF-α induction of core 2 GlcNAc-T. Human leukocytes(U937 cells) were exposed to human recombinant TNF-alpha (8pg/ml) in the presence and absence of test compounds After 24h incubation, the activity of core 2 GlcNAc-T was measured, and expressed as pmoles/h/mg protein (iii). Cell free assay of core 2 GlcNAc-T in cell free assays of core 2 GlcNAc-T Heart lysates from either from TNF- alpha over expressing transgenic mice (female, B6.SJL-Tg (TNF) supplied by Taconic-M+B, Bomholtveg 10, 8680 35 Ry, Denmark) or from BB rats (Festing 1979) was exposed to various concentrations of test compound for 1h at 37°C. Activity of core 2 GlcNAc-T was measured, and expressed as pmoles/h/mg protein.

1c. Measurement of core 2 GlcNAc-T activity

40 [0088] To measure core 2 GlcNAc-T activity, leukocytes were washed in PES, frozen and lysed in 0.9% Triton X-100 at 0°C. The activity of core 2 GlcNAc-T was measured as described below. Cell free assays are preformed by substituting heart lysates for cell lysates. [0089] The reaction was performed in a reaction mixture containing 50 mM 2(N-morpholino)ethanesulfonic acid (MES, Sigma, Dorset, UK), pH 7.0, 1 mM UDP-6 [’H]-N-acetylglucosamine (16,000 dpm/nmol, NEN Life Science Products, 45 Hounslow, UK), 0.1 M GlcNAc (Sigma, Dorset, OK), 1 mM Galβ1-3GalNAcα-p-nitrophenol (Sigma, Dorset, UK) as substrate, and 16 ml of lysate (100-200 mg protein) for a final volume of 32 ml. After incubating the mixture for 1 hour at 37°C, the reaction was terminated with 1 ml of ice-cold distilled water and processed on a C18 Sep-Pak column (Waters- Millipore, Watford, UK). After washing the column with 20 ml of distilled water, the product was eluted with 5 ml of methanol. The radioactivity of the samples was counted in a liquid scintillationβ -counter (LKB-Wallac, London, UK). 50 Endogenous activity of core 2 GlcNAc-T was measured in the absence of the added acceptor. The specific activity was expressed as pmoles/h/mg of cell protein. In each case, the protein concentration was determined with BioRad protein assay (BioRad, Hertfordshire, UK). Results are shown in table 5.

Table 5: Approximate Ic50 values (nM) for example compounds 55 Compound Number Cell free assay Cell based assay Trigoneoside IVa 6 0.9* 75

14 EP 1 909 802 B1

(continued)

Compound Number Cell free assay Cell based assay Glycoside F 7 5** b 5 Shatavarin I 21 1* 0.75 Shatavarin IV 22 c** † ***Protogracillin 26 3* 0.25 10 * Cell freeassays were carried out onheart lysates ofTNF- α mice as described above. ** Cell free assays were carried out on heart lysates of BB rats as described above. a 33% inhibition at 20nM 15 b 100% inhibition at 22nM n BB rat heart lysate c 89% inhibition at 22nM in BB rat heart lysate † no activity detected at 22.5 nM *** comparative compound

20 [0090] Compound C (at 20ng/ml) was found to inhibit Core 2 GlcNAc-T approximately 98.5% compared to controls, in TNF-α treated Human leukocytes (U937 cells). The sample of compound C was approximately 82.5% pure by HPLC at 205nM [0091] The approximate IC50 of Trigoneoside IVa was found to be between 0.25nM and 0.9nM in cell free systems. Further analysis of a sample prepared according to applicants co pending WO05/060977 indicates that it contains 25 approximately 7.5% protodioscin and 9% Trigonelloside C (Yoshikawa et al., Heterocycles 47, 397-405 (1998)).

[0092] The IC50 of Glycoside F was found to be approximately 5nM. Further analysis of the preparation indicates that it contains a small amount of Trigonelloside C. [0093] The IC50 of Protodioscin (93.3% pure) produced as described in applicants co pending WO05/060977 was found to be approximately 20nM. The sample contained 1.5% Trigoneoside IVa. A sample prepared fromTribulus 30 terrestris (Chromodex Inc. 2952 S. Daimler Street Santa Ana CA 92705), which was 97% pure, and had an NMR spectrum consistent with protodioscin, appeared to demonstrate no activity at concentrations of 50mM. Thus Trigoneoside IVa activity could account for at least some of the activity seen in the protodioscin sample prepared as per WO05/060977. [0094] Trigonelloside C is similar to Protodioscin but is the opposite isomer at carbon 25. A preparation of this compound according to co pending WO05/060977 was 98.2% pure and contained no measurable quantity of other Core 2 GlcNAc- 35 T inhibitors. A preparation of Trigonelloside C prepared according to WO05/060977 inhibited Core 2 GlcNAc-T 69% at 2.5nM.

REFERENCES

40 [0095]

1. Hu K. et al Planta Medica, 63(2), 161-165 (1997). 2. Dong M. et al., Planta Med. 67(9):853-7 (2001). 3. Yoshikawa M. et al., Heterocycles 47, 397-405 (1998). 45 4. Akhov L. S. et al., J. Agric. Food Chem. 47(8), 3193-3196 (1999) 5. Mimaki Y. et al., Chem Pharm Bull (Tokyo). 46(11):1829-32 (1998). 6. Shimomura H. et al., Phytochemistry 28, 3163-3170 (1989). 7. Kawasaki T. et al., Chemical & Pharmaceutical Bulletin, 22(9), 2164-75 (1974). 8. Ori K. et al. Phytochemistry. 31(8):2767-75 (1992). 50 9. Mimaki Y. et al Phytochemistry 37(1):227-32 (1994). 10. Nakamura O. et al., Phytochemistry. 36(2):463-7 (1994). 11. Mimaki Y and Sasheda Y. Chem. Pharm. Bull. 38(11), 3055-9(1990). 12. Sashida Y. et al., Chem. Pharm. Bull. 39(9), 2362-8(1991) 13. Haladova M. et al., Pharmazie, 54(2), 159-160 (1999). 55 14. Sharma et al., Phytochemistry. 33(3):683-6. (1993). 15. Petit G. et al., Journal of natural products 54, 1491-1502. 16. Vasil’eva I. S. et al., Appl. Biochem. Microbiol. 31, 206-209 (1995).

15 EP 1 909 802 B1

17. Vasil’eva I. et al., Prikl Biokhim Mikrobiol. 20(3):404-6 (1984). 18. Joshi J. et al., Indian J. Chem. 27B, 12-16 (1988). 19. Ravikumar P. R. et al., Indian J. Chem. 26B, 1012-1017 (1987).

5 Claims

1. A compound of the formula (III) or a pharmaceutically acceptable salt, ether or ester thereof for use in the treatment of a condition associated with detrimental activity of the enzyme core 2 GlcNAc-T selected from the group consisting 10 of ischemia reperfusion injury, restenosis and thrombosis

20 Wherein the "A" is glucose; and Z is a steroid moiety of the formula (VI)

35 wherein:

R7 and R14 are independently selected from H and -OH; 8 R is C1-6 alkyl; 9 11 16 R , R , and R are independently selected from H and C 1-6 alkyl; 40 R10 is H or -OH or the H normally present is absent and R 10 is =O; 12 R is H or -OH or C1-6 alkyl; R13 is H; 15 13 15 R is H, -OH or C1-6 alkyl or R and R taken together form a -CH2-CH2-group; 17 R is H, C1-6 alkyl or C1-6 hydroxyalkyl; 45 ------represents bond that is either double or single;

and wherein; the steroid moiety Z incorporates a further group selected from the groups VIa and VIc:

16 EP 1 909 802 B1

wherein: 15 18 23 R and R are independently selected from C1-6 alkyl; R19 and R24 are independently selected from H and -OH; 20 19 20 R is H, -OH or C 1-6 alkoxy or R and R taken together represent the second bond of a double bound joining adjacent carbon atoms; 20 21 R is C1-6 alkyl, C2-6 alkenyl, or C2-6 alkynyl or a C1-6 alkyl or C2-6 alkenyl group substituted by one or more groups selected from the group consisting of -OH, C 1-6 alkoxy and Sac 4; 25 R is C1-6 alkyl, C1-6 hydroxyalkyl, or = CH2; Sac 4 is a saccharide group; and X is either O or NH. 25 2. A compound or a pharmaceutically acceptable salt, ether, or ester thereof for use according to claim 1 wherein the compoundis a 6-deoxy- α-L-mannopyranosyl-(1→2)-O-[β-D-glucopyranosyl-(1→4)]-β-D-Glucopyranosideof steroid Z.

30 3. A compound or a pharmaceutically acceptable salt, ether, or ester thereof for use according to claim 1 or 2 wherein 8 9 the steroid group Z is a group of the formula VI incorporating the further group VIa wherein R is -CH3 and R is H 11 12 16 17 21 and R is -CH3; R is H or -OH; R and R are H and R is 3-methyl but-2-enyl, 2-methyl-prop-2-enyl, 3- methylenebutyl substituted at the 4-position by Sac 4, 1-hydroxy-3-methylbutanyl substituted at the 4-position by Sac4, 3-methylenebutyl substituted at the 4-position by Sac 4, or 1-methoxy-3-methylbutanyl substituted at the 4- 35 position by Sac 4.

4. A compound or a pharmaceutically acceptable salt, ether, or ester thereof for use according to claim 1 or 2 wherein the compound is of the formula III in which the steroid group Z is selected from group VI which incorporate the further 7 9 10 13 14 16 17 19 12 8 11 18 group VIa and in which R , R , R , R , R , R , R , and R are H; R is H or -OH; R , R and R are -CH3; 40 15 20 21 R is H, -OH; R is -OH or O CH3; and R is 4-hydroxy-3-methylbutanyl, 3-methylenebutyl substituted at the 4- position by Glc, 1-hydroxy-3-methylbutanyl substituted at the 4-position by Glc or 1-methoxy-3-methylbutanyl substituted at the 4-position by Glc.

5. A compound or a pharmaceutically acceptable salt, ether, or ester thereof for use according to claim 1 or 2 wherein 45 9 11 the steroid group Z is a group of the formula VI incorporating the further group VIa wherein R is -CH3; R is H; 20 21 10 15 16 17 R is H; R is a C2-6 alkenyl; R is H; R is -OH or -CH2-CH2-; and R and R are -CH3.

6. A compound or a pharmaceutically acceptable salt, ether, or ester thereof for use according to claim 1 or 2 wherein 8 9 the steroid group Z is a group of the formula VI incorporating the further group Vlc wherein R is -CH3, and R is H 50 11 12 15 16 17 and R is -CH3; R is H or -OH; R is H or -OH; R and R are H and X is O.

7. A compound or a pharmaceutically acceptable salt, ether, or ester thereof for use according to claim 1 to 3 in which Sac 4 is a monosaccharide.

55 8. A compound or a pharmaceutically acceptable salt, ether, or ester thereof for use according to claim 7 in which Sac 4 is glucose.

9. A compound or a pharmaceutically acceptable salt, ether, or ester thereof for use according to claims 1 or 2 wherein

17 EP 1 909 802 B1

the steroid group Z is selected from the group consisting of the radicals of sarsasapogenin, smilagenin, 12 β-hyde- oxysmilagenin, rhodeasapogenin, isorhodiasapogenin, samogenin, 12 β-hydroxysamogenin, markogenin, yonogenin, convallagenin A, convallagenin B, tokorogenin, tigogenin, neotigogenin, gitogenin, agigenin, digitogenin, chlorogenin, paniculogenion, (25R)-spirostan-3β, 17α,21-triol, allogenin, (25R)-5α-spirostan-2α,3β,5α,6α-tetraol, 5 (24S,25R)-5α-spirostan-2α,3β,5α,6β,24-pentaol, yamogenin, diogenin, yuccagenin, liligenin, ruscogenin, (25S)-ruscogenin, neopraserigenin, pennogenin, isonuatigenin, cepagenin, 24α-hydroxypennogenin, ophiogenin, sibiricogenin, convallamarogenin, neoruscogenin, hecogenin, neohecogenin, manogenin, sisalagenin and hispigenin.

10 10. A compound or a pharmaceutically acceptable salt, ether, or ester thereof for use according to claims 1 or 2 wherein the steroid group Z is selected from the group consisting of the radicals of solasodine, soladulcidine, tomatidine, and 5-dehydrotomatidine.

11. A compound or a pharmaceutically acceptable salt, ether, or ester thereof for use according to claim 1 wherein the 15 steroid group Z is selected from the group consisting of:

18 EP 1 909 802 B1

55 12. A compound or a pharmaceutically acceptable salt, ether, or ester thereof for use according to claims 1 or 2 wherein the steroid group Z is selected from the group consisting of:

19 EP 1 909 802 B1

20 EP 1 909 802 B1

30 13. A compound or a pharmaceutically acceptable salt, ether, or ester thereof for use according to claims 1 or 2 wherein the compound is selected from the group consisting of: Trigoneoside Iva, which is (3β,25S)-26-(β-D-glucopyranosyloxy)-22-hydroxyfurost-5-en-3-yl O-6-deoxy-α-L-mannopyranosyl-(1→2)-O-[β-D-glucopyranosyl-(1→4)]-β-D- glucopyranoside, glycoside F, which is (3β,25R)-26-(β-D-glucopyranosyloxy)-22- hydroxyfurost-5-en-3-yl O-6-deoxy-α-L-mannopyranosyl-(1→2)-O-[β-D-glucopyranosyl- (1→4)]-β-D-glucopyranoside, Pardarinoside C which is 35 (3β,5α,22α,25R)-26-(acetyloxy)-14,17- dihydroxy-22-methoxyfurostan-3-yl O-6-deoxy-α-L-mannopyranosyl- (1→2)-O-[β-D-glucopyranosyl-(1→4)]-β-D-Glucopyranoside, Pardarinoside D which isβ -D-glucopyranoside, (3β,5α,22α,25R)-26-(acetyloxy)-17-hydroxy-22-methoxyfurostan-3-yl O-6-deoxy-α-L-mannopyranosyl-(1→2)-O-[β-D-glucopyranosyl-(1→4)]-β-D-glucopyranoside, Balanitin VI which is (3 β,25S)-spirost-5-en-3-yl-O- α-L-rhamnopyranosyl-(1→2)-O-[β-D-glucopyranosyl-β-D-glucopyranoside, Deltonin which is (3β,25R)-spirost-5- 40 en-3-yl-O-α-L-rhamnopyranosyl-(1→2)-O-[β-D-glucopyranosyl-β-D- glucopyranoside, Shatavarin I which is (3β,5β,22α,25S)-26-(β-D-glucopyranosyloxy)-22- hydroxyfurostan-3-yl α O-6-deoxy--L-mannopyranosyl-(1→2)-O-[β-D-glucopyranosyl- (1→4)]-β-D-glucopyranoside, Shatavarin IV which is (3 β,5β,25S)-spirostan-3-yl O-6-deoxy-α-L-mannopyranosyl-(1→2)-O-[β-D-glucopyranosyl-β-D-glucopyranoside, Compound 8 which is β,25R)-26-( (3 β-D-glucopyranosyloxyl-22-methoxyfurost-5-en-3-yl O-6-deoxy-α-L-man- 45 nopyranosyl-(1→2)-O-[β-D-glucopyranosyl-(1→4)]-β-D-Gluco-pyranoside, compound 12 which is [(3β,12α,25R)- 12-hydroxyspirostan-3-yl O-6-deoxy-α-L-mannopyranosyl-(1→2)-[β-D-glucopyranosyl-(1→4)]-β-D-glucopyranoside], compound 13 which is [(25S)-spirost-5-ene-3β27-diol 3-O-{6-deoxy-α-L-mannopyranosyl-(1→2)--[β-D-glucopyranosyl-(1→4)]-β-D-glucopyranoside}], compound 14 which is [(25R, 26R)-26-methoxyspirost-5-en-3β-ol 3- O-{6-deoxy-α-L-mannopyranosyl-(1→2)-[β-D-glucopyranosyl-(1→4)]-β-D-glucopyranoside}], compound 15 which 50 is [3β,25R,27(S)]-27-(4-carboxy-3-hydroxy-3-methyl1-oxobutoxy)spirost-5-en-3-yl O-6-deoxy-α-L-mannopyranosyl-(1→2)-O-[β-D-glucopyranosyl-(1→4)]-β-D-glucopyranoside], compound 16 which is [3β,25R,27(S)]-27-[(3-hydroxy-5-methoxy-3-methyl-1,5-dioxopentyl)oxy]spirost-5-en-3-yl O-6-deoxy-α-L-mannopyranosyl-(1→2)-O-[β-D- glucopyranosyl-(1→4)]-β-D-glucopyranoside], compound 17 which is β-D-Glucopyranoside, (3β,25R,26R)-17-hydroxy-26-methoxyspirost-5-en-3-yl O-6-deoxy-α-L-mannopyranosyl-(1→2)-O-[β-D-glucopyranosyl-(1→4)]-β-D- 55 glucopyranoside, compound 18 which is (3β,25R,26R)-26-hydroxyspirost-5-en-3-yl O-6-deoxy-α-L-mannopyranosyl-(1→2)-O-[β-D-glucopyranosyl-(1→4)]-β-D-glucopyranoside, compound 23 which is 26-O-β-D-glucopyranosylnuatigenin 3-O-α-L-rhamnopyranosyl- (1→2)-O-[β-D-glucopyranosyl-(1→4)]-β-D-glucopyranoside, compound 25 which is β (3,25S)-26-(β-D-gluco-pyranosyloxy)-22-hydroxyfurost-5, 25 (27) dien-3-yl O-6-deoxy-α-L-man-

21 EP 1 909 802 B1

nopyranosyl-(1→2)-O-[β-D-glucopyranosyl-(1→4)]-β-D-glucopyranoside, and compound 26 which is solasodine 3-O-α-L-rhamnopyranosyl-(1→2)-O-[β-D-glucopyranosyl-(1→4)]-β-D-glucopyranoside.

14. A compound or a pharmaceutically acceptable salt, ether, or ester thereof for use according to claims 1 or 2 wherein 5 the compound is selected from Trigoneoside IVa, Glycoside F, Deltonin, Balanitin VI, Shatavarin I, Shatavarin IV, and Compound 25.

15. A compound or a pharmaceutically acceptable salt, ether, or ester thereof for use according to any one of claims 1 to 14 wherein the isolated compound is incorporated into a food or beverage product. 10 16. A compound or a pharmaceutically acceptable salt, ether, or ester thereof for use according to any one of claims 1 to 15 wherein the compound is purified from a natural source.

17. A compound or a pharmaceutically acceptable salt, ether, or ester thereof for use according to any one of claims 1 15 to 15 wherein the compound is synthetic.

18. A compound or a pharmaceutically acceptable salt, ether, or ester thereof for use according to Claim 16 wherein the compound is at least 1 % pure.

20 19. A compound or a pharmaceutically acceptable salt, ether, or ester thereof for use according to Claim 16 wherein the compound is at least 30% pure.

20. A compound or a pharmaceutically acceptable salt, ether, or ester thereof for use according to Claim 16 wherein the compound is at least 50% pure. 25 21. A compound or a pharmaceutically acceptable salt, ether, or ester thereof for use according to Claim 16 wherein the compound is at least 80% pure.

22. A compound or a pharmaceutically acceptable salt, ether, or ester thereof for use according to Claim 16 wherein 30 the compound is at least 90% pure.

23. A compound or a pharmaceutically acceptable salt, ether, or ester thereof for use according to Claim 16 wherein the compound is at least 95% pure.

35 24. A compound or a pharmaceutically acceptable salt, ether, or ester thereof for use according to any one of claims 1 to 5, 7, 8, 11, or 13 to 23 which is a furostanol compound of the formula VIa in which the carbon at the 22-position of the furostanol structure, is substituted by methoxy.

25. A compound or a pharmaceutically acceptable salt, ether, or ester thereof for use according to any of the preceding 40 claims by the transdermal, airway, rectal, vaginal or topical route.

26. A compound or a pharmaceutically acceptable salt, ether, or ester thereof for use according to any of the preceding claims by the oral route.

45 27. A compound or a pharmaceutically acceptable salt, ether, or ester thereof for use according to claim 1 by the parenteral route.

Patentansprüche 50 1. EineVerbindung derFormel (III) oder einpharmazeutisch verträgliches Salz, Ether oderEster davon zur Verwendung in der Behandlung eines Zustandes, der mit einer schädlichen Aktivität des Enzyms Core-2-GlcNAc-T assoziiert ist und aus der Gruppe ausgewählt ist, bestehend aus Ischämie-Reperfusionsverletzung, Restenose und Thrombose

22 EP 1 909 802 B1

10 worin "A" Glucose ist; und Z eine Steroid-Komponente der Formel (VI) ist:

worin: 25 R7 und R14 unabhängig voneinander aus H und -OH ausgewählt sind; 8 R C1-6-Alkyl ist; 9 11 16 R , R und R unabhängig voneinander aus H und C 1-6-Alkyl ausgewählt sind; R10 H oder -OH ist oder das normalerweise vorhandene H fehlt und R 10 =O ist; 30 12 R H oder -OH oder C1-6-Alkyl ist; R13 H ist; 15 13 15 R H, -OH oder C1-6-Alkyl ist oder R und R zusammengenommen eine -CH2-CH2- Gruppe bilden; 17 R H, C1-6-Alkyl oder C1-6-Hydroxyalkyl ist; ----- für eine Bindung steht, die entweder eine Doppel- oder Einfachbindung ist; 35 und worin; die Steroid-Komponente Z eine weitere Gruppe enthält, ausgewählt aus den Gruppen VIa und VIc:

50 worin:

18 23 R und R unabhängig voneinander aus C1-6-Alkyl ausgewählt sind; R19 und R24 unabhängig voneinander aus H und -OH ausgewählt sind; 55 20 19 20 R H, -OH oder C1-6-Alkoxy ist oder R und R zusammengenommen für die zweite Bindung einer Doppel- bindung stehen, die benachbarte Kohlenstoffatome miteinander verbindet; 21 R C1-6-Alkyl, C2-6-Alkenyl oder C2-6-Alkinyl oder eine C1-6-Alkyl- oder C2-6-Alkenylgruppe ist, die mit einer oder mehreren Gruppen substituiert ist, ausgewählt aus der Gruppe, bestehend aus -OH, C 1-6-Alkoxy und Sac 4;

23 EP 1 909 802 B1

25 R C1-6-Alkyl, C1-6-Hydroxyalkyl oder =CH2 ist; Sac 4 eine Saccharid-Gruppe ist; und X entweder O oder NH ist.

5 2. Eine Verbindung oder ein pharmazeutisch verträgliches Salz, Ether oder Ester davon zur Verwendung gemäß Anspruch 1, wobei die Verbindung ein 6-Desoxy-α-L-mannopyranosyl-(1→2)-O-[β-D-glucopyranosyl-(1→4)]-β-D- glucopyranosid von Steroid Z ist.

3. Eine Verbindung oder ein pharmazeutisch verträgliches Salz, Ether oder Ester davon zur Verwendung gemäß 10 Anspruch 1 oder 2, wobei die Steroid-Gruppe Z eine Gruppe der Formel VI ist, die die weitere Gruppe VIa enthält, 8 9 11 12 16 17 21 worin R -CH3 ist und R H ist und R -CH3 ist; R H oder -OH ist, R und R H sind und R 3-Methylbut-2- enyl, 2-Methyl-prop-2-enyl, 3-Methylenbutyl, das an der 4-Position mit Sac 4 substituiert ist, 1-Hydroxy-3-methylbutanyl, das an der 4-Position mit Sac 4 substituiert ist, 3-Methylenbutyl, das an der 4-Position mit Sac 4 substituiert ist, oder 1-Methoxy-3-methylbutanyl ist, das an der 4-Position mit Sac 4 substituiert ist. 15 4. Eine Verbindung oder ein pharmazeutisch verträgliches Salz, Ether oder Ester davon zur Verwendung gemäß Anspruch 1 oder 2, wobei die Verbindung der Formel III, in der die Steroid-Gruppe Z aus Gruppe VI ausgewählt ist, die die weitere Gruppe VIa enthält und in der R7, R9, R10, R13, R14, R16, R17 und R19 H sind; R12 H oder -OH ist; 8 11 18 15 20 21 R , R und R -CH3 sind; R H oder -OH ist; R -OH oder -OCH3 ist und R 4-Hydroxy-3-methylbutanyl, 3- 20 Methylenbutyl, das an der 4-Position mit Glc substituiert ist, 1-Hydroxy-3-methylbutanyl, das an der 4-Position mit Glc substituiert ist, oder 1-Methoxy-3-methylbutanyl ist, das an der 4-Position mit Glc substituiert ist.

5. Eine Verbindung oder ein pharmazeutisch verträgliches Salz, Ether oder Ester davon zur Verwendung gemäß Anspruch 1 oder 2, wobei die Steroid-Gruppe Z eine Gruppe der Formel VI ist, die die weitere Gruppe VIa enthält, 25 9 11 20 21 10 15 16 worin R -CH3 ist; R H ist; R H ist; R ein C2-6-Alkenyl ist; R H ist; R -OH oder -CH2-CH2- ist; und R und 17 R -CH3 sind.

6. Eine Verbindung oder ein pharmazeutisch verträgliches Salz, Ether oder Ester davon zur Verwendung gemäß Anspruch 1 oder 2, wobei die Steroid-Gruppe Z eine Gruppe der Formel VI ist, die die weitere Gruppe VIc enthält, 30 8 9 11 12 15 16 17 worin R -CH3 ist, und R H ist und R -CH 3 ist; R H oder -OH ist; R H oder -OH ist; R und R H sind und X O ist.

7. Eine Verbindung oder ein pharmazeutisch verträgliches Salz, Ether oder Ester davon zur Verwendung gemäß Anspruch 1 bis 3, wobei Sac 4 ein Monosaccharid ist.

35 8. Eine Verbindung oder ein pharmazeutisch verträgliches Salz, Ether oder Ester davon zur Verwendung gemäß Anspruch 7, wobei Sac 4 Glucose ist.

9. Eine Verbindung oder ein pharmazeutisch verträgliches Salz, Ether oder Ester davon zur Verwendung gemäß Anspruch 1 oder 2, wobei die Steroid-Gruppe Z aus der Gruppe ausgewählt ist, bestehend aus den Resten von 40 Sarsasapogenin, Smilagenin, 12β-Hydroxysmilagenin, Rhodeasapogenin, Isorhodeasapogenin, Samogenin, 12β- Hydroxysamogenin, Markogenin, Yonogenin, Convallagenin A, Convallagenin B, Tokorogenin, Tigogenin, Neoti- gogenin, Gitogenin, Agigenin, Digitogenin, Chlorogenin, Paniculogenion, (25R)-Spirostan-3 β, 17α,21-Triol, Alloge- nin, (25R)-5α-Spirostan-2α,3β,5α,6α-tetraol, (24S,25R)-5α-Spirostan-2α,3β,5a,6β,24-pentaol, Yamogenin, Dioge- nin, Yuccagenin, Liligenin, Ruscogenin, (25S)-Ruscogenin, Neopraserigenin, Pennogenin, Isonuatigenin, Cepage- 45 nin, 24α-Hydroxypennogenin, Ophiogenin, Sibiricogenin, Convallamarogenin, Neoruscogenin, Hecogenin, Neohe- cogenin, Manogenin, Sisalagenin und Hispigenin.

10. Eine Verbindung oder ein pharmazeutisch verträgliches Salz, Ether oder Ester davon zur Verwendung gemäß Anspruch 1 oder 2, wobei die Steroid-Gruppe Z aus der Gruppe ausgewählt ist, bestehend aus den Resten von 50 Solasodin, Soladulcidin, Tomatidin und 5-Dehydrotomatidin.

11. Eine Verbindung oder ein pharmazeutisch verträgliches Salz, Ether oder Ester davon zur Verwendung gemäß Anspruch 1, wobei die Steroid-Gruppe Z aus der Gruppe ausgewählt ist, bestehend aus:

24 EP 1 909 802 B1

25 EP 1 909 802 B1

40 12. Eine Verbindung oder ein pharmazeutisch verträgliches Salz, Ether oder Ester davon zur Verwendung gemäß Anspruch 1 oder 2, wobei die Steroid-Gruppe Z aus der Gruppe ausgewählt ist, bestehend aus:

26 EP 1 909 802 B1

27 EP 1 909 802 B1

13. Eine Verbindung oder ein pharmazeutisch verträgliches Salz, Ether oder Ester davon zur Verwendung gemäß 15 Anspruch 1 oder 2, wobei die Verbindung aus der Gruppe ausgewählt ist, bestehend aus:Trigoneosid IVa, das (3β,25S)-26-(β-D-Glucopyranosyloxy)-22-hydroxyfurost-5-en-3-yl-O-6-desoxy-α-L-mannopyranosyl-(1→2)-O-[β- D-glucopyranosyl-(1→4)]-β-D-gluco-pyranosid ist, Glycosid F, das (3β,25R)-26-(β-D-Glucopyranosyloxy)-22-hydroxyfurost-5-en-3-yl-O-6-desoxy-α-L-mannopyranosyl-(1→2)-O-[β-D-glucopyranosyl-(1→4)]-β-D-glucopyranosid ist, Pardarinosid C, das (3β,5α,22α,25R)-26-(Acetyloxy)-14,17-dihydroxy-22-methoxyfurostan-3-yl-O-6-deso- 20 xy-α-L-mannopyranosyl-(1→2)-O-[β-D-glucopyranosyl-(1→4)]-β-D-glucopyranosid ist, Pardarinosid D, das β-D- Glucopyranosid, (3β,5α,22α,25R)-26-(Acetyloxy)-17-hydroxy-22-methoxyfurostan-3-yl-O-6-desoxy-α-L-mannopyranosyl-(1→2)-O-[β-D-glucopyranosyl-(1→4)]-β-D-glucopyranosid ist, Balanitin VI, das (3 β,25S)-Spirost-5-en-3-yl- O-α-L-rhamnopyranosyl-(1→2)-O-[β-D-glucopyranosyl-β-D-glucopyranosid ist, Deltonin, das (3β,25R)-Spirost-5- en-3-yl-O-α-L-rhamnopyranosyl-(1→2)-O-[β-D-glucopyranosyl-β-D-glucopyranosid ist, Shatavarin I, das 25 (3β,5β,22α,25S)-26-(β-D-Glucopyranosyloxy)-22-hydroxyfurostan-3-yl-O-6-desoxy-α-L-mannopyranosyl-(1→2)-O-[β-D-glucopyranosyl-(1→4)]-β-D-glucopyranosid ist, Shatavarin IV, das (3β,5β,25S)-Spirostan-3-yl- O-6-desoxy-α-L-mannopyranosyl-(1→2)-O-[β-D-glucopyranosyl-β-D-glucopyranosid ist, Verbindung 8, die (3β,25R)-26-(β-D-Glucopyranosyloxy)-22-methoxyfurost-5-en-3-yl-O-6-desoxy-α-L-mannopyranosyl-(1→2)-O-[β- D-glucopyranosyl-(1→4)]-β-D-glucopyranosid ist, Verbindung 12, die [(3β,12α,25R)-12-Hydroxyspirostan-3-yl-O- 30 6-desoxy-α-L-mannopyranosyl-(1→2)-[β-D-glucopyranosyl-(1→4)]-β-D-glucopyranosid] ist, Verbindung 13, die [(25S)-Spirost-5-en-3β,27-diol-3-O-{6-desoxy-α-L-mannopyranosyl-(1→2)-[β-D-glucopyranosyl-(1→4)]-β-D-glucopyranosid}] ist, Verbindung 14, die [(25R,26R)-26-Methoxyspirost-5-en-3β -ol-3-O-{6-desoxy-α-L-mannopyranosyl-(1-2)-[β-D-glucopyranosyl-(1→4)]-β-D-glucopyranosid}] ist, Verbindung 15, die [3β,25R,27(S)]-27-(4-Carboxy- 3-hydroxy-3-methyl-1-oxobutoxy)spirost-5-en-3-yl-O-6-desoxy-α-L-mannopyranosyl-(1→2)-O-[β-D-glucopyrano- 35 syl-(1→4)]-β-D-glucopyranosid] ist, Verbindung 16, die [3β,25R,27(S)]-27-[(3-Hydroxy-5-methoxy-3-methyl-1,5- dioxopentyl)oxy]spirost-5-en-3-yl-O-6-desoxy-α-L-mannopyranosyl-(1→2)-O-[β-D-glucopyranosyl-(1→4)]-β-D- glucopyranosid] ist, Verbindung 17, die β-D-Glucanopyranosid, (3β,25R,26R)-17-Hydroxy-26-methoxyspirost-5- en-3-yl-O-6-desoxy-α-L-mannopyranosyl-(1→2)-O-[β-D-glucopyranosyl-(1→4)]-β-D-glucopyranosid ist, Verbin- dung 18, die (3β,25R,26R)-26-Hydroxyspirost-5-en-3-yl-O-6-desoxy-α-L-mannopyranosyl-(1→2)-O-[β-D-glucopy- 40 ranosyl-(1→4)]-β-D-glucopyranosid ist, Verbindung 23, die 26-O-β-D-Glucopyranosylnuatigenin-3-O-α-L-rhamnopyranosyl-(1→2)-O-[β-D-glucopyranosyl-(1→4)]-β-D-glucopyranosid ist, Verbindung 25, die (3 β,25S)-26-(β-D-Glu- copyrosyloxy)-22-hydroxyfurost-5,25(27)-dien-3-yl-O-6-desoxy-α-L-mannopyrano-syl-(1→2)-O-[β-D-glucopyranosyl-(1→4)]-β-D-glucopyranosid ist, und Verbindung 26, die Solasodin 3-O-α-L-rhamnopyranosyl-(1→2)-O-[β-D- glucopyranosyl-(1→4)]-β-D-glucopyranosid ist. 45 14. Eine Verbindung oder ein pharmazeutisch verträgliches Salz, Ether oder Ester davon zur Verwendung gemäß Anspruch 1 oder 2, wobei die Verbindung aus Trigoneosid IVa, Glycosid F, Deltonin, Balanitin VI, Shatavarin I, Shatavarin IV und Verbindung 25 ausgewählt ist.

50 15. Eine Verbindung oder ein pharmazeutisch verträgliches Salz, Ether oder Ester davon zur Verwendung gemäß einem der Ansprüche 1 bis 14, wobei die isolierte Verbindung in einem Nahrungsmittel- oder Getränkeprodukt beigemengt ist.

16. Eine Verbindung oder ein pharmazeutisch verträgliches Salz, Ether oder Ester davon zur Verwendung gemäß einem 55 der Ansprüche 1 bis 15, wobei die Verbindung aus einer natürlichen Quelle gereinigt ist.

17. Eine Verbindung oder ein pharmazeutisch verträgliches Salz, Ether oder Ester davon zur Verwendung gemäß einem der Ansprüche 1 bis 15, wobei die Verbindung eine synthetische Verbindung ist.

28 EP 1 909 802 B1

18. Eine Verbindung oder ein pharmazeutisch verträgliches Salz, Ether oder Ester davon zur Verwendung gemäß Anspruch 16, wobei die Verbindung wenigstens eine 1%ige Reinheit aufweist.

19. Eine Verbindung oder ein pharmazeutisch verträgliches Salz, Ether oder Ester davon zur Verwendung gemäß 5 Anspruch 16, wobei die Verbindung wenigstens eine 30%ige Reinheit aufweist.

20. Eine Verbindung oder ein pharmazeutisch verträgliches Salz, Ether oder Ester davon zur Verwendung gemäß Anspruch 16, wobei die Verbindung wenigstens eine 50%ige Reinheit aufweist.

10 21. Eine Verbindung oder ein pharmazeutisch verträgliches Salz, Ether oder Ester davon zur Verwendung gemäß Anspruch 16, wobei die Verbindung wenigstens eine 80%ige Reinheit aufweist.

22. Eine Verbindung oder ein pharmazeutisch verträgliches Salz, Ether oder Ester davon zur Verwendung gemäß Anspruch 16, wobei die Verbindung wenigstens eine 90%ige Reinheit aufweist. 15 23. Eine Verbindung oder ein pharmazeutisch verträgliches Salz, Ether oder Ester davon zur Verwendung gemäß Anspruch 16, wobei die Verbindung wenigstens eine 95%ige Reinheit aufweist.

24. Eine Verbindung oder ein pharmazeutisch verträgliches Salz, Ether oder Ester davon zur Verwendung gemäß einem 20 der Ansprüche 1 bis 5, 7, 8, 11 oder 13 bis 23, die eine Furostanol-Verbindung der Formel VIa ist, in der der Kohlenstoff an der 22-Position der Furostanol-Struktur mit Methoxy substituiert ist.

25. Eine Verbindung oder ein pharmazeutisch verträgliches Salz, Ether oder Ester davon zur Verwendung gemäß einem der vorhergehenden Ansprüche über den transdermalen, rektalen, vaginalen, topischen Weg oder Atemweg. 25 26. Eine Verbindung oder ein pharmazeutisch verträgliches Salz, Ether oder Ester davon zur Verwendung gemäß einem der vorhergehenden Ansprüche über den oralen Weg.

27. Eine Verbindung oder ein pharmazeutisch verträgliches Salz, Ether oder Ester davon zur Verwendung gemäß 30 Anspruch 1 über den parenteralen Weg.

Revendications

35 1. Composé de formule (III) ou sel, éther ou ester de celui-ci, acceptable sur le plan pharmaceutique, pour une utilisation dans le traitement d’un état, associé à une activité préjudiciable de l’enzyme core 2 GlcNAc-T, choisi parmi le groupe constitué des blessures de reperfusion ischémie, resténose et thrombose

Dans lequel « A » est un glucose et Z est un groupe fonctionnel stéroïde de formule (VI) :

29 EP 1 909 802 B1

dans lequel : 15 R7 et R14 sont d’une manière indépendante choisis entre H et -OH ; 8 R est un alkyle C1-6 ; 9 11 16 R , R et R sont d’une manière indépendante choisis entre H et un alkyle C1-6 ; R10 est H ou -OH ou le H normalement présent est absent et R10 est = 0 ; 20 12 R est H ou -OH ou un alkyle C1-6 ; R13 est H ; 15 13 15 R est H, -OH ou un alkyle C1-6 ; ou R et R pris ensemble forment un groupe -CH2-CH2-; 17 R est H, un alkyle C1-6 ; ou un hydroxyalkyle C1-6; ----- représente une liaison qui est soit double, soit simple ; 25 et dans lequel le groupe fonctionnel stéroïde Z incorpore un autre groupe choisi parmi les groupes VIa et VIc :

dans lesquels :

45 18 23 R et R sont d’une manière indépendante choisis parmi des alkyles C1-6 ; R19 et R24 sont d’une manière indépendante choisis entre H et -OH ; 20 19 20 R est H, -OH ou un alkoxy C1-6 ; ou R et R pris ensemble représentent la seconde liaison d’une double liaison liant des atomes de carbone adjacents ; 21 R est un alkyle C1-6, un alkényle C2-6, ou un alkynyle C2-6 ou un groupe alkyle C1-6 ; ou alkényle C2-6 objet 50 d’une substitution par un ou plusieurs groupes choisis parmi le groupe constitué de -OH, un alkoxy C1-6 ; et Sac 4 ; 25 R est un alkyle C1-6, un hydroxyalkyle C1-6, ou =CH2 ; Sac 4 est un groupe saccharide ; et X est soit O, soit NH.

55 2. Composé ou sel, éther ou ester de celui-ci, acceptable sur le plan pharmaceutique, pour une utilisation conforme à la revendication 1, dans lequel le composé est un 6-désoxy-α-L-mannopyranosyl-(1→2)-O-[β-D-glucopyranosyl-(1→4]-β-D-glucopyranoside du stéroïde Z.

30 EP 1 909 802 B1

3. Composé ou sel, éther ou ester de celui-ci, acceptable sur le plan pharmaceutique, pour une utilisation conforme à la revendication 1 ou 2, dans lequel le groupe stéroïde Z est un groupe de formule VI incorporant l’autre groupe 8 9 11 12 16 17 21 VIa dans lequel R est -CH3 et R est H et R est -CH3 ; R est H ou -OH ; R et R sont H et R est un 3- méthyl but-2-ényl, 2-méthyl-prop-2-ényl, 3-méthylènebutyle offrant une substitution en position 4 par un Sac 4, 1- 5 hydroxy-3-méthylbutanyle offrant une substitution en position 4 par un Sac 4, 3-méthylènebutyle offrant une substitution en position 4 par un Sac 4, ou 1-méthoxy-3-méthylbutanyle offrant une substitution en position 4 par un Sac 4.

4. Composé ou sel, éther ou ester de celui-ci, acceptable sur le plan pharmaceutique, pour une utilisation conforme à la revendication 1 ou 2 dans lequel le composé est de formule III dans laquelle le groupe stéroïde Z est choisi 10 dans le groupe VI qui incorpore l’autre groupe VIa et dans lequel R7, R9, R10, R13, R14, R16, R17 et R19 sont H ; R12 8 11 18 15 20 21 est H ou -OH ; R , R et R sont -CH3 ; R est H, -OH ; R est -OH ou OCH- 3 et R est un 4-hydroxy-3- méthylbutanyle, 3-méthylènebutyle offrant une substitution en position 4 par Glc, 1-hydroxy-3-méthylbutanyle offrant une substitution en position 4 par Glc ou 1-méthoxy-3-méthylbutanyle offrant une substitution en position 4 par Glc.

15 5. Composé ou sel, éther ou ester de celui-ci, acceptable sur le plan pharmaceutique, pour une utilisation conforme à la revendication 1 ou 2, dans lequel le groupe stéroïde Z est un groupe de formule VI incorporant l’autre groupe 9 11 20 21 10 15 VIa dans lequel R est -CH3, R est H ; R est H ; R est un alkényle C2-6, R est H ; R est -OH ou -CH2-CH2-; 16 17 et R et R sont -CH3.

20 6. Composé ou sel, éther ou ester de celui-ci, acceptable sur le plan pharmaceutique, pour une utilisation conforme à la revendication 1 ou 2, dans lequel le groupe stéroïde Z est un groupe de formule VI incorporant l’autre groupe 8 9 11 12 15 16 17 VIc dans lequel R est -CH3, et R est H et R est -CH3 ; R est H ou -OH ; R est H ou -OH ; R et R sont H et X est O.

25 7. Composé ou sel, éther ou ester de celui-ci, acceptable sur le plan pharmaceutique, pour une utilisation conforme à la revendication 1 à 3, dans lequel Sac 4 est un monosaccharide.

8. Composé ou sel, éther ou ester de celui-ci, acceptable sur le plan pharmaceutique, pour une utilisation conforme à la revendication 7, dans lequel Sac 4 est un glucose. 30 9. Composé ou sel, éther ou ester de celui-ci, acceptable sur le plan pharmaceutique, pour une utilisation conforme aux revendications 1 ou 2, dans lequel le groupe stéroïde Z est choisi dans le groupe constitué des radicaux de sarsasapogénine, smilagénine, 12 β-hydésoxysmilagénine, rhodéasapogénine, isorhodiasapogénine, samogénine, 12β-hydroxysamogénine, markogénine, yonogénine, convallagénine A, convallagénine B, tokorogénine, tigogénine, 35 néotigogénine, gitogénine, agigénine, digitogénine, chlorogénine, paniculogénion, (25R)-spirostan-3 β, 17α,21-triol, allogénine, (25R)-5α-spirostan-2α,3β,5α,6α-tétraol, (24S,25R)-5α-spirostan-2α,3β,5α,6β,24-pentaol, yamogénine, diogénine, yuccagénine, liligénine, ruscogénine, (25S)-ruscogénine, néoprasérigénine, pennogénine, isonuatigé- nine, cépagénine, 24α-hydroxypennogénine, ophiogénine, sibiricogénine, convallamarogénine, néoruscogénine, hécogénine, néohécogénine, manogénine, sisalagénine et hispigénine. 40 10. Composé ou sel, éther ou ester de celui-ci, acceptable sur le plan pharmaceutique, pour une utilisation conforme aux revendications 1 ou 2, dans lequel le groupe stéroïde Z est choisi dans le groupe constitué des radicaux de solasodine, soladulcidine, tomatidine, et 5-déshydrotomatidine.

45 11. Composé ou sel, éther ou ester de celui-ci, acceptable sur le plan pharmaceutique, pour une utilisation conforme à la revendication 1, dans lequel le groupe stéroïde Z est choisi dans le groupe constitué des :

31 EP 1 909 802 B1

32 EP 1 909 802 B1

12. Composé ou sel, éther ou ester de celui-ci, acceptable sur le plan pharmaceutique, pour une utilisation conforme aux revendications 1 ou 2, dans lequel le groupe stéroïde Z est choisi dans le groupe constitué des : 30

33 EP 1 909 802 B1

13. Composé ou sel, éther ou ester de celui-ci, acceptable sur le plan pharmaceutique, pour une utilisation conforme 55 aux revendications 1 ou 2, dans lequel le composé est choisi dans le groupe constitué des : Trigoneoside Iva qui est ,25S)-26-( (3β β-D-glucopyranosyloxy)-22-hydroxyfurost-5-ène-3-yl O-6-désoxy-α-L-mannopyranosyl-(1→2)-O-[β-D-glucopyranosyl-(1→4)]-β-D-glucopyranoside, glycoside F qui est (3β,25R)-26-(β-D-glucopyranosyloxy)-22-hydroxyfurost-5-ène-3-yl O-6-désoxy-α-L-mannopyranosyl-(1→2)-O-[β-D-glucopyranosyl-(1→4)]-β-

34 EP 1 909 802 B1

D-glucopyranoside, Pardarinoside C qui est (3β,5α,22α,25R)-26-(acétyloxy)-14,17-dihydroxy-22-méthoxyfuros- tan-3-yl O-6-désoxy- α-L-mannopyranosyl-(1→2)-O-[β-D-glucopyranosyl-(1-4)]-β-D-glucopyranoside,Pardarinosi- de D qui est β-D-glucopyranoside, (3β,5α,22α,25R)-26-(acétyloxy)-17-hydroxy-22-méthoxyfurostan-3-yl O-6-dé- soxy-α-L-mannopyranosyl-(1→2)-O-[β-D-glucopyranosyl-(1→4)]-β-D-glucopyranoside, Balanitine VI qui est 5 (3β,25S)-spirost-5-ène-3-yl-O-α-L-rhamnopyranosyl-(1→2)-O-[β-D-glucopyranosyl-β-D-glucopyranoside, Deltoni- ne qui est (3β,25R)-spirost-5-ène-3-yl-O-α-L-rhamnopyranosyl-(1→2)-O-[β-D-glucopyranosyl-β-D-glucopyranoside, Shatavarine I qui est (3β,5β,22α,25S)-26-(β-D-glucopyranosyloxy)-22-hydroxyfurostan-3-yl O-6-désoxy-α-L- mannopyranosyl-(1→2)-O-[β-D-glucopyranosyl-(1→4)]-β-D-glucopyranoside, Shatavarine IV qui est (3β,5β,25S)-spirostan-3-yl O-6-désoxy-α-L-mannopyranosyl-(1-2)-O-[β-D-glucopyranosyl-β-D-glucopyranoside, 10 Composé 8 qui est (3β ,25R)-26-(β-D-glucopyranosyloxy)-22-méthoxyfurost-5-ène-3-yl O-6-désoxy-α-L-mannopyranosyl-(1→2)-O-[β-D-glucopyranosyl-(1→4)]-β-D-glucopyranoside, composé 12 qui est [(3β, 12α, 25R)-12-hydroxyspirostan-3-yl O-6-désoxy-α-L-mannopyranosyl-(1→2)-[β-D-glucopyranosyl-(1→4)]-β-D-glucopyranoside], composé 13 qui est [(25S)-spirost-5-ène-3 β27-diol 3-O-{6-désoxy-α-L-mannopyranosyl-(1→2)-[β-D-gluco-pyranosyl-(1→4)]-β-D-glucopyranoside}], composé 14 qui est [(25R,26R)-26-méthoxyspirost-5-ène-3 β-ol 3-O-{6-désoxy- 15 α-L-mannopyranosyl-(1→2)-[β-D-glucopyranosyl-(1→4)]-β-D-glucopyranoside}], composé 15 qui est [3β,25R,27(S)]-27-(4-carboxy-3-hydroxy-3-méthyl-1-oxobutoxy)spirost-5-ène-3-yl O-6-désoxy-α-L-mannopyranosyl-(1→2)-O-[β-D-glucopyranosyl-(1→4)]-β-D-glucopyranoside], composé 16 qui est [3,25R,27(S)]-27-[(3-hy-β droxy-5-méthoxy-3-méthyl-1,5-dioxopentyl)oxy]spirost-5-ène-3-yl O-6-désoxy-α-L-mannopyranosyl-(1→2)-O-[β- D-glucopyranosyl-(1→4)]-β-D-glucopyranoside], composé 17 qui est β-D-glucopyranoside, (3β,25R,26R)-1,7-hy- 20 droxy-26-méthoxyspirost-5-ène-3-yl O-6-désoxy-α-L-mannopyranosyl-(1→2)-O-[β-D-glucopyranosyl-(1→4)]-β-D- glucopyranoside, composé 18 qui est (3β,25R,26R)-26-hydroxyspirost-5-ène-3-yl O-6-désoxy-α-L-mannopyranosyl-(1→ 2)-O-[β-D-glucopyranosyl-(1→4)]-β-D-glucopyranoside, composé 23 qui est 26-O- β-D-glucopyranosylnua- tigénine 3-O-α-L-rhamnopyranosyl-(1→2)-O-[β-D-glucopyranosyl-(1→4)]-β-D-glucopyranoside, composé 25 qui est (3β,25S)-26-(β-D-glucopyranosyloxy)-22-hydroxyfurost-5, 25(27) diène-3-yl O-6-désoxy-α-L-mannopyrano- 25 syl-(1→2)-O-[β-D-glucopyranosyl-(1→4)]-β-D-glucopyranoside, et composé 26 qui est solasodine 3-O-α-L-rhamnopyranosyl-(1→2)-O-[β-D-glucopyranosyl-(1→4)]-β-D-glucopyranoside.

14. Composé ou sel, éther ou ester de celui-ci, acceptable sur le plan pharmaceutique, pour une utilisation conforme aux revendications 1 ou 2, dans lequel le composé est choisi parmi les Trigoneoside Iva, Glycoside F, Deltonine, 30 Balanitine VI, Shatavarine I, Shatavarine IV et le Composé 25.

15. Composé ou sel, éther ou ester de celui-ci, acceptable sur le plan pharmaceutique, pour une utilisation conforme à l’une quelconque des revendications 1 à 14, dans lequel le composé isolé est incorporé dans un produit alimentaire ou boisson. 35 16. Composé ou sel, éther ou ester de celui-ci, acceptable sur le plan pharmaceutique, pour une utilisation conforme à l’une quelconque des revendications 1 à 15, dans lequel le composé est purifié à partir d’une source naturelle.

17. Composé ou sel, éther ou ester de celui-ci, acceptable sur le plan pharmaceutique, pour une utilisation conforme 40 à l’une quelconque des revendications 1 à 15, dans lequel le composé est synthétique.

18. Composé ou sel, éther ou ester de celui-ci, acceptable sur le plan pharmaceutique, pour une utilisation conforme à la revendication 16, dans lequel le composé est pur à au moins 1%.

45 19. Composé ou sel, éther ou ester de celui-ci, acceptable sur le plan pharmaceutique, pour une utilisation conforme à la revendication 16, dans lequel le composé est pur à au moins 30%.

20. Composé ou sel, éther ou ester de celui-ci, acceptable sur le plan pharmaceutique, pour une utilisation conforme à la revendication 16, dans lequel le composé est pur à au moins 50%. 50 21. Composé ou sel, éther ou ester de celui-ci, acceptable sur le plan pharmaceutique, pour une utilisation conforme à la revendication 16, dans lequel le composé est pur à au moins 80%.

22. Composé ou sel, éther ou ester de celui-ci, acceptable sur le plan pharmaceutique, pour une utilisation conforme 55 à la revendication 16, dans lequel le composé est pur à au moins 90%.

23. Composé ou sel, éther ou ester de celui-ci, acceptable sur le plan pharmaceutique, pour une utilisation conforme à la revendication 16, dans lequel le composé est pur à au moins 95%.

35 EP 1 909 802 B1

24. Composé ou sel, éther ou ester de celui-ci, acceptable sur le plan pharmaceutique, pour une utilisation conforme à l’une quelconque des revendications 1 à 5, 7, 8, 11, ou 13 à 23, qui est un composé furostanol de formule VIa dans laquelle le carbone en position 22 de la structure furostanol est l’objet d’une substitution par un méthoxy.

5 25. Composé ou sel, éther ou ester de celui-ci, acceptable sur le plan pharmaceutique, pour une utilisation, conforme à l’une quelconque des revendications précédentes, par la voie transcutanée, aérienne, rectale, vaginale ou topique.

26. Composé ou sel, éther ou ester de celui-ci, acceptable sur le plan pharmaceutique, pour une utilisation, conforme à l’une quelconque des revendications précédentes, par la voie orale. 10 27. Composé ou sel, éther ou ester de celui-ci, acceptable sur le plan pharmaceutique, pour une utilisation conforme à la revendication 1, par la voie parentérale.

36 EP 1 909 802 B1

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REFERENCES CITED IN THE DESCRIPTION

This list of references cited by the applicant is for the reader’s convenience only. It does not form part of the European patent document. Even though great care has been taken in compiling the references, errors or omissions cannot be excluded and the EPO disclaims all liability in this regard.

Patent documents cited in the description

• WO 0187548 A [0002] • US 6593301 B [0004] • WO 05060977 A [0003] [0057] [0084] [0091] [0093] • CN 1586493 A [0005] [0094] • CN 1415625 A [0005]

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