Saccharide Derivative of a Toxic Payload and Antibody Conjugates Thereof

Abstract

A molecule comprising a saccharide bound via an O-glycosidic bond to a hydroxyl group of a toxic payload molecule is disclosed. An antibody-drug conjugate comprising an antibody covalently bound to a toxic payload molecule, optionally via a linker group, and a saccharide bound via an O-glycosidic bond to a hydroxyl group of the toxic payload molecule is further disclosed.

1 - 63 . (canceled) 
     
     
         64 . An antibody-drug conjugate comprising an antibody covalently bound to a toxic payload molecule, optionally via a linker group, and a saccharide bound via an O-glycosidic bond to a hydroxyl group of the toxic payload molecule,
 wherein the saccharide is optionally bound via an O-glycosidic bond to an ester linker group bound via an ester bond to a hydroxyl group of the toxic payload molecule, and wherein the ester linker group has a structure according to formula XII   
       
         
           
           
               
               
           
         
         wherein X is the bond to the toxic payload molecule; R1′ is the saccharide bound via the O-glycosidic bond; and RE is selected from the group consisting of —CH 2 —, —CH 2 CH 2 —, —CH 2 CH 2 CH 2 —, —CH 2 CH 2 CH 2 CH 2 —, C 1 -C 4  alkyl, heteroalkyl, branched alkyl, branched heteroalkyl, cyclic alkyl, or cyclic heteroalkyl, and substituted C 1 -C 4  alkyl, heteroalkyl, branched alkyl, branched heteroalkyl, cyclic alkyl, and cyclic heteroalkyl. 
       
     
     
         65 . The antibody-drug conjugate according to  claim 64  represented by formula VI
   AB-[L-A-R1] n    Formula VI
 
 wherein 
 AB is an antibody; 
 L is absent, a linker group, or L′-R2; 
 A is a toxic payload molecule; 
 R1 is a saccharide bound via an O-glycosidic bond to a hydroxyl group of the toxic payload molecule, or L″-R1′, wherein R1′ is a saccharide bound via an O-glycosidic bond to L″, and L″ is bound via an ester bond to a hydroxyl group of the toxic payload molecule and has a structure according to formula XII; R2 is a saccharide bound via an O-glycosidic bond to a hydroxyl group of the toxic payload molecule; 
 L′ is a linker group and the saccharide R2 is bound to the linker group L′ via a covalent bond; and 
 n is at least 1, 
 or 
 the antibody-drug conjugate according to  claim 64  represented by formula XI
   AB-[L′-R2-A] n    Formula XI
 
 
 wherein 
 AB is an antibody; 
 L′ is absent or a linker group; 
 A is an toxic payload molecule; 
 
       R2 is a saccharide bound via an O-glycosidic bond to a hydroxyl group of the toxic payload molecule; and
 n is at least 1. 
 
     
     
         66 . The antibody-drug conjugate according to  claim 64  represented by a formula selected from the group consisting of formulas VII, VII′, and VII″: 
       
         
           
           
               
               
           
         
         wherein 
         AB is an antibody; 
         L is absent or a linker group; 
         R1 is a saccharide bound via an O-glycosidic bond to the toxic payload molecule, or L″-R1′, wherein R1′ is a saccharide bound via an O-glycosidic bond to L″, and L″ is bound via an ester bond to a hydroxyl group of the toxic payload molecule and has a structure according to formula XII; 
         AA is a peptide comprising a hydroxyl group whereto R1 is bound via the O-glycosidic bond; and 
         n is at least 1; 
         or 
         the antibody-drug conjugate according to  claim 64  or  65  represented by a formula selected from the group consisting or formulas XI′, XI″, XI′″, or XI″″: 
       
       
         
           
           
               
               
           
         
         wherein 
         AB is an antibody; 
         L′ is absent or a linker group; 
         A is an toxic payload molecule; 
         R2 is a saccharide bound via an O-glycosidic bond to a hydroxyl group of the toxic payload molecule; and 
         n is at least 1. 
       
     
     
         67 . The antibody-drug conjugate according to  claim 64 , wherein R1 or R1′ comprises or is a monosaccharide selected from the group consisting of β-D-galactose, N-acetyl-β-D-galactosamine, N-acetyl-α-D-galactosamine, N-acetyl-β-D-glucosamine, β-D-glucuronic acid, α-L-iduronic acid, α-D-galactose, α-D-glucose, β-D-glucose, α-D-mannose, β-D-mannose, α-L-fucose, β-D-xylose, neuraminic acid and any analogue or modification thereof, and the monosaccharide is bound to the toxic payload molecule via the O-glycosidic bond. 
     
     
         68 . The antibody-drug conjugate according to the  claim 67 , wherein the modification is sulfate, phosphate, carboxyl, amino, or O-acetyl modification of the monosaccharide. 
     
     
         69 . The antibody-drug conjugate according to  claim 64 , wherein the R1 or R1′ comprises or is a disaccharide. 
     
     
         70 . The antibody-drug conjugate according to  claim 64 , wherein the O-glycosidic bond is hydrolysable by a lysosomal or an intracellular glycohydrolase. 
     
     
         71 . The antibody-drug conjugate according to  claim 64 , wherein the antibody is selected from the group consisting of bevacizumab, tositumomab, etanercept, trastuzumab, adalimumab, alemtuzumab, gemtuzumab ozogamicin, efalizumab, rituximab, infliximab, abciximab, basiliximab, palivizumab, omalizumab, daclizumab, cetuximab, panitumumab, epratuzumab, 2G12, lintuzumab, nimotuzumab and ibritumomab tiuxetan, or the antibody is selected from the group consisting of an anti-EGFR1 antibody, an epidermal growth factor receptor 2 (HER2/neu) antibody, an anti-CD22 antibody, an anti-CD30 antibody, an anti-CD33 antibody, and an anti-CD20 antibody. 
     
     
         72 . The antibody-drug conjugate according to  claim 64 , wherein n is in the range of 1 to about 20, or 1 to about 15, or 1 to about 10, or 2 to 10, or 2 to 6, or 2 to 5, or 2 to 4; 3 to about 20, or 3 to about 15, or 3 to about 10, or 3 to about 9, or 3 to about 8, or 3 to about 7, or 3 to about 6, or 3 to 5, or 3 to 4; 4 to about 20, or 4 to about 15, or 4 to about 10, or 4 to about 9, or 4 to about 8, or 4 to about 7, or 4 to about 6, or 4 to 5; or n is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20. 
     
     
         73 . The antibody-drug conjugate according to  claim 64 , wherein the linker group comprises a peptide and optionally a self-immolative group linking the peptide and the toxic payload molecule. 
     
     
         74 . The antibody-drug conjugate according to  claim 64 , wherein the linker group comprises a saccharide bound to a hydroxyl group of the toxic payload molecule via an O-glycosidic bond. 
     
     
         75 . The antibody-drug conjugate according to  claim 74 , wherein L is L′-R2, wherein R2 is a saccharide bound via an O-glycosidic bond to a hydroxyl group of the toxic payload molecule, L′ is a linker group, and the saccharide R2 is bound to the linker group L′ via a covalent bond. 
     
     
         76 . The antibody-drug conjugate according to  claim 64 , wherein the antibody-drug conjugate comprises or is a conjugate selected from the group consisting of the following:
 a conjugate of O-β-D-galactopyranosylmonomethylauristatin E (MMAG) and antibody;   a conjugate of Val-Cit-PAB-MMAG and antibody;   a conjugate of N-(6-azido-6-deoxy-D-galactosyl)-MMAG and antibody;   a conjugate of O—(N-acetylneuraminyl-β-D-galactopyranosyl)-MMAE (MMAS) and antibody;   a conjugate of Val-Cit-PAB-MMAS and antibody;   a conjugate of N-(6-azido-6-deoxy-D-galactosyl)-MMAS and antibody;   a conjugate of MMAG and anti-EGFR antibody;   a conjugate of MMAS and anti-EGFR antibody;   a conjugate of β-D-4′-demethylepipodophyllotoxin β-D-4-galactopyranosylglucopyranoside (Galβ1,4Glc-PT) and antibody;   a conjugate of NeuAcα2,6Galβ1,4Glc-PT and antibody;   a conjugate of α2,6-N-acetyl-9-azidoneuraminyl-β-D-4-galactopyranos-ylglucopyranoside (9-azido-NeuAcα2,6Galβ1,4Glc-PT) and antibody;   a conjugate of 9-azido-NeuAcα2,6Galβ1,4Glc-PT and DBCO-modified antibody;   a conjugate of galactopyranosyl-DM1 and antibody;   a conjugate of α2,6-NeuAc-galactopyranosyl-DM1 and antibody;   a conjugate of 9-azido-sialyl derivative of galactopyranosyl-DM1 and antibody;   a conjugate of 9-azido-sialyl derivative of galactopyranosyl-DM1 and DBCO-modified antibody;   a conjugate of dexamethasone 21-O-beta-D-galactopyranoside (Gal-Dexa) and antibody;   a conjugate of NeuAcα2,6Gal-Dexa and antibody;   a conjugate of 9N3NeuAcα2,6Gal-Dexa and antibody;   a conjugate of 9N3NeuAcα2,6Gal-Dexa and DBCO-modified antibody;   a conjugate of β-1,4-galactosylated Xyl-taxol (Galβ1,4Xyl-Taxol) and antibody;   a conjugate of NeuAcα2,6Galβ1,4Xyl-Taxol and antibody;   a conjugate of 9-N3-NeuAcα2,6Galβ1,4Xyl-Taxol and antibody;   a conjugate of 9-N3-NeuAcα2,6Galβ1,4Xyl-Taxol and DBCO-modified antibody;   a conjugate of N-(6-azido-6-deoxy-D-galactosyl)-MMAF-IleVal(GlcNAcβ-)Ser and antibody;   a conjugate of N-(6-azido-6-deoxy-D-galactosyl)-MMAF-IleVal(GlcNAcβ-)Ser and DBCO-modified antibody;   a conjugate of VaCitPAB-MMAF-IleVal(GlcNAcβ)Ser and antibody;   a conjugate of dimeric duocarmycin f-D-galactopyranoside and antibody;   a conjugate of dimeric duocarmycin sialylgalactoside and antibody;   a conjugate of 9-azido-modified dimeric duocarmycin sialylgalactoside and antibody;   a conjugate of 9-azido-modified dimeric duocarmycin sialylgalactoside and DBCO-modified antibody;   a conjugate of duocarmycin β-D-galactopyranoside and antibody;   a conjugate of duocarmycin sialylgalactoside and antibody;   a conjugate of Val-Cit-Gly-duocarmycin sialylgalactoside and antibody;   a conjugate of Neu5Acα2,6Gal-duocarmycin and antibody;   a conjugate of Gal-Duocarmycin-PAB-CV-DBCO and antibody;   a conjugate of Neu5Acα2,6MMAG and antibody;   a conjugate of Neu5Acα2,3MMAG and antibody;   a conjugate of maleimidocaproyl-VC-PAB-MMAG and antibody;   a conjugate of beta-D-Galactopyranosyl-(glycolic acid ester)-duocarmycin and antibody;   a conjugate of sialyl-beta-D-Galactopyranosyl-(glycolic acid ester)-duocarmycin and antibody;   a conjugate of O-β-D-galactopyranosylmonomethylauristatin E (MMAG) and trastuzumab, cetuximab, brentuximab or nimotuzumab;   a conjugate of Val-Cit-PAB-MMAG and trastuzumab, cetuximab, brentuximab or nimotuzumab;   a conjugate of O—(N-acetylneuraminyl-β-D-galactopyranosyl)-MMAE (MMAS) and trastuzumab, cetuximab, brentuximab or nimotuzumab;   a conjugate of Val-Cit-PAB-MMAS and trastuzumab, cetuximab, brentuximab or nimotuzumab;   a conjugate of duocarmycin β-D-galactopyranoside and trastuzumab, cetuximab, brentuximab or nimotuzumab;   a conjugate of duocarmycin sialylgalactoside and trastuzumab, cetuximab, brentuximab or nimotuzumab;   a conjugate of Val-Cit-Gly-duocarmycin sialylgalactoside and trastuzumab, cetuximab, brentuximab or nimotuzumab;   a conjugate of beta-D-Galactopyranosyl-(glycolic acid ester)-duocarmycin and trastuzumab, cetuximab, brentuximab or nimotuzumab;   a conjugate of sialyl-beta-D-Galactopyranosyl-(glycolic acid ester)-duocarmycin and trastuzumab, cetuximab, brentuximab or nimotuzumab;   a conjugate of beta-D-Galactopyranosyl-(glycolic acid ester)-MMAE and antibody;   a conjugate of glucuronyl-duocarmycin and antibody;   a conjugate of 2-amino-2-deoxy-β-D-glucopyranosyl duocarmycin and antibody;   a conjugate of 2-acylamido-2-deoxy-β-D-glucopyranosyl duocarmycin and antibody;   a conjugate of beta-glucuronyl-monomethylauristatin E (MMAU) and antibody;   a conjugate of maleimidocaproyl-VC-PAB-MMAU and antibody;   a conjugate of MMAU-PAB-CV-maleimidoyl and antibody;   a conjugate of beta-glucosyl-monomethylauristatin E (MMAX) and antibody;   a conjugate of maleimidocaproyl-VC-PAB-MMAX and antibody;   a conjugate of MMAX-PAB-CV-maleimidoyl and antibody;   a conjugate of MMAG-PAB-CV-maleimidoyl and antibody;   a conjugate of 2-acetamido-2-deoxy-β-D-glucopyranosyl duocarmycin and antibody;   a conjugate of Gal-duocarmycin and antibody;   a conjugate of Gal-duocarmycin-PAB-CV-maleimidoyl and antibody;   a conjugate of glycosyl-maytansinoid and antibody;   a conjugate of glycosyl-N-glucosylmaytansinoid and antibody;   a conjugate of Gal-N-glucosylmaytansinoid and antibody;   a conjugate of NeuAc-Gal-N-glucosylmaytansinoid and antibody;   a conjugate of glycosyl-tubulysin and antibody;   a conjugate of Gal-tubulysin and antibody;   a conjugate of NeuAc-Gal-tubulysin and antibody;   a conjugate of glycosyl-α-amanitin and antibody;   a conjugate of Gal-α-amanitin and antibody;   a conjugate of NeuAc-Gal-α-amanitin and antibody;   a conjugate of glycosyl-cryptophycin and antibody;   a conjugate of Gal-cryptophycin and antibody; and,   a conjugate of NeuAc-Gal-cryptophycin and antibody.   
     
     
         77 . The antibody-drug conjugate according to  claim 64 , wherein the glycoside-to-antibody ratio is in the range of 1 to about 100, or 1 to about 60, or 1 to about 20, or 1 to about 10; 2 to about 40, or 2 to about 20, or 2 to about 12, or 2 to about 10, or 2 to about 8, or 2 to about 6, or 2 to about 4; 3 to about 60, or 3 to about 40, or 3 to about 30, or 3 to about 20, or 3 to about 15, or 3 to about 10, or 3 to about 8, or 3 to about 6, or 3 to 4; 4 to about 60, or 4 to about 40, or 4 to about 20, or 4 to about 16, or 4 to about 12, or 4 to about 10, or 4 to about 8, or 4 to about 7, or 4 to about 6, or 4 to 5; over 3, over 4, over 5, over 6, over 7, over 8, over 9, over 10, over 11, over 12, over 13, over 14, over 15, or over 16; or the glycoside-to-antibody ratio is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 14, 15, 16, 18, 20, 21, 22, 24, 25, 26, 27, 28, 30, 32, 34, 35, 36, 38, 40, 42, 44, 45, 46, 48, 50, 52, 54, 55, 56, 58, 60, 62, 63, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, or more than 100. 
     
     
         78 . A molecule comprising a saccharide bound via an O-glycosidic bond to a hydroxyl group of a toxic payload molecule,
 wherein the saccharide is optionally bound via an O-glycosidic bond to an ester linker group bound via an ester bond to a hydroxyl group of the toxic payload molecule, and wherein the ester linker group has a structure according to formula XII   
       
         
           
           
               
               
           
         
         wherein X is the bond to the toxic payload molecule; R1′ is the saccharide bound via the O-glycosidic bond; and RE is selected from the group consisting of —CH 2 —, —CH 2 CH 2 —, —CH 2 CH 2 CH 2 —, —CH 2 CH 2 CH 2 CH 2 —, C 1 -C 4  alkyl, heteroalkyl, branched alkyl, branched heteroalkyl, cyclic alkyl, or cyclic heteroalkyl, and substituted C 1 -C 4  alkyl, heteroalkyl, branched alkyl, branched heteroalkyl, cyclic alkyl, and cyclic heteroalkyl. 
       
     
     
         79 . The molecule according to  claim 78 , wherein the molecule is represented by formula I
   R2-A-R1   Formula I
   wherein A is a toxic payload molecule;   R1 is a saccharide bound to a hydroxyl group of the toxic payload molecule via an O-glycosidic bond, or L″-R1′, wherein R1′ is a saccharide bound via an O-glycosidic bond to L″, and L″ is bound via an ester bond to a hydroxyl group of the toxic payload molecule and has a structure according to formula XII; and   R2 is H, a linker group, or a saccharide bound to a hydroxyl group of the toxic payload molecule via an O-glycosidic bond.   
     
     
         80 . The molecule according to  claim 78  represented by Formula II, Formula II′, Formula II″, or Formula II′″: 
       
         
           
           
               
               
           
         
         wherein R1 is a saccharide bound to the toxic payload molecule via an O-glycosidic bond, or L″-R1′, wherein R1′ is a saccharide bound via an O-glycosidic bond to L″, and L″ is bound via an ester bond to a hydroxyl group of the toxic payload molecule and has a structure according to formula XII; 
         AA is a peptide comprising a hydroxyl group whereto R1 is bound via the O-glycosidic bond; and 
         R2 is H or a linker group. 
       
     
     
         81 . The molecule according to  claim 80 , wherein the saccharide R1 or R1′ comprises or is a monosaccharide selected from the group consisting of β-D-galactose, N-acetyl-β-D-galactosamine, N-acetyl-α-D-galactosamine, N-acetyl-β-D-glucosamine, β-D-glucuronic acid, α-L-iduronic acid, α-D-galactose, α-D-glucose, β-D-glucose, α-D-mannose, β-D-mannose, α-L-fucose, β-D-xylose, neuraminic acid and any analogue or modification thereof, and the monosaccharide is bound to the toxic payload molecule via the O-glycosidic bond. 
     
     
         82 . The molecule according to  claim 78 , wherein the linker group comprises a saccharide bound to a hydroxyl group of the toxic payload molecule via an O-glycosidic bond. 
     
     
         83 . The molecule according to  claim 78 , wherein the molecule comprises or is a molecule selected from the group consisting of the following:
 O-β-D-galactopyranosylmonomethylauristatin E (MMAG);   Val-Cit-PAB-MMAG;   N-(6-azido-6-deoxy-D-galactosyl)-MMAG;   O—(N-acetylneuraminyl-β-D-galactopyranosyl)-MMAE (MMAS);   Val-Cit-PAB-MMAS;   N-(6-azido-6-deoxy-D-galactosyl)-MMAS;   β-D-4′-demethylepipodophyllotoxin;   β-D-4-galactopyranosylglucopyranoside (Galβ1,4Glc-PT);   α2,6-N-acetylneuraminyl-β-D-4-galactopyranosylgluco-pyranoside (NeuAcα2,6Galβ1,4Glc-PT);   α2,6-N-acetyl-9-azidoneuraminyl-β-D-4-galactopyranos-ylglucopyranoside (9-azido-NeuAcα2,6Galβ1,4Glc-PT);   galactopyranosyl-DM1;   α2,6-NeuAc-galactopyranosyl-DM1;   9-azido-sialyl derivative of galactopyranosyl-DM1;   dexamethasone 21-O-beta-D-galactopyranoside (Gal-Dexa);   NeuAcα2,6Gal-Dexa;   9N3NeuAcα2,6Gal-Dexa;   β-1,4-galactosylated Xyl-taxol (Galβ1,4Xyl-Taxol);   NeuAcα2,6Galβ1,4Xyl-Taxol;   9-N3-NeuAcα2,6Galβ1,4Xyl-Taxol;   N-(6-azido-6-deoxy-D-galactosyl)-MMAF-IleVal(GlcNAcβ-)Ser;   ValCitPAB-MMAF-IleVal(GlcNAcβ)Ser,   dimeric duocarmycin β-D-galactopyranoside;   dimeric duocarmycin sialylgalactoside;   duocarmycin β-D-galactopyranoside;   duocarmycin sialylgalactoside;   Val-Cit-Gly-duocarmycin sialylgalactoside;   Neu5Acα2,6Gal-Duocarmycin;   Gal-Duocarmycin-PAB-CV-DBCO;   Neu5Acα2,6MMAG;   Neu5Acα2,3MMAG;   maleimidocaproyl-VC-PAB-MMAG;   beta-D-Galactopyranosyl-(glycolic acid ester)-duocarmycin;   sialyl-beta-D-Galactopyranosyl-(glycolic acid ester)-duocarmycin;   beta-D-Galactopyranosyl-(glycolic acid ester)-MMAE;   2-amino-2-deoxy-β-D-glucopyranosyl duocarmycin;   beta-glucuronyl-monomethylauristatin E (MMAU);   beta-glucosyl-monomethylauristatin E (MMAX);   2-acetamido-2-deoxy-β-D-glucopyranosyl duocarmycin;   2-acylamido-2-deoxy-β-D-glucopyranosyl duocarmycin;   Gal-duocarmycin;   glycosyl-maytansinoid;   glycosyl-N-glucosylmaytansinoid;   Gal-N-glucosylmaytansinoid;   NeuAc-Gal-N-glucosylmaytansinoid;   glycosyl-tubulysin;   Gal-tubulysin;   NeuAc-Gal-tubulysin;   glycosyl-α-amanitin;   Gal-α-amanitin;   NeuAc-Gal-α-amanitin;   glycosyl-cryptophycin;   Gal-cryptophycin; and   NeuAc-Gal-cryptophycin.   
     
     
         84 . A pharmaceutical composition comprising the antibody-drug conjugate according to  claim 64 . 
     
     
         85 . A method of treating and/or modulating the growth of and/or prophylaxis of tumor cells in humans or animals, wherein the pharmaceutical composition according to  claim 84  is administered to a human or animal in an effective amount. 
     
     
         86 . The method according to  claim 85 , wherein the tumor cells are selected from the group consisting of leukemia cells, lymphoma cells, breast cancer cells, prostate cancer cells, ovarian cancer cells, colorectal cancer cells, gastric cancer cells, squamous cancer cells, small-cell lung cancer cells, head-and-neck cancer cells, multidrug resistant cancer cells, and testicular cancer cells.