Modified glycoprotein, protein-conjugate and process for the preparation thereof

Abstract

The invention relates to a glycoprotein comprising an optionally fucosylated glycan according to formula (105) or (106), wherein Su(A) x is a modified sugar moiety comprising one or more functional groups A. Functional group A is independently selected from the group consisting of a thiol group, a halogen, a sulfonyloxy group, a halogenated acetamido group, a mercaptoacetamido group and a sulfonated hydroxyacetamido group. The invention also relates to a glycoprotein-conjugate wherein a glycoprotein according to the invention is conjugated to a molecule of interest. Said molecule of interest may for example be an active substance. The invention further relates to a process for the preparation of a modified glycoprotein, and to a method for the preparation of a glycoprotein-conjugate. The invention particularly relates to modified antibodies, antibody-conjugates, antibody-drug conjugates and methods for the preparation thereof.

1 - 22 . (canceled) 
     
     
         23 . A method of treating a disease, adverse affect attributable to the disease, or a combination thereof comprising administering to a subject in need thereof an effective amount of a glycoprotein-conjugate comprising the formula:
   Pr[glycan-SuR[(Q) p -S-Z-L(D) r ] x ] y      wherein:   Pr represents a protein;   glycan comprising the formula (101A) or (102A):   
       
         
           
           
               
               
           
         
          wherein G is a monosaccharide, or a linear or branched oligosaccharide comprising 2 to 20 sugar moieties;
 b is 0 or 1; 
 d is 0 or 1; and 
 e is 0 or 1; 
 
         Su is a sugar or sugar derivative; 
         Q is —N(H)C(O)CH 2 — or —CH 2 —; 
         Z is a connecting group selected from -succimide-, —CH 2 —C(O)—N(R 9 )— and —C(═CH)—CH 2 —C(O)—N(R 9 )—, wherein the carbon atom is connected to S and the nitrogen atom is connected to L;
 wherein R 9  is selected from the group consisting of L(D) r , hydrogen, C 1 -C 24  alkyl groups, C 6 -C 24  aryl groups, C 7 -C 24  alkylaryl groups and C 7 -C 24  arylalkyl groups, wherein said C 1 -C 24  alkyl groups, C 6 -C 24  aryl groups, C 7 -C 24  alkylaryl groups and C 7 -C 24  arylalkyl groups are optionally substituted; 
 
         L is a linker; 
         D is a molecule of interest; 
         p is 0 or 1; 
         r is 1 to 20; 
         x is 1, 2, 3 or 4; and 
         y is 1 to 20; or 
       
       a pharmaceutical composition comprising the effective amount of the glycoprotein-conjugate and a pharmaceutically acceptable excipient, wherein the effective amount is effective for treating the disease, the adverse affect attributable to the disease, or a combination thereof in the subject. 
     
     
         24 . The method of  claim 23 , wherein the subject is a mammal. 
     
     
         25 . The method of  claim 24 , wherein the mammal is a human. 
     
     
         26 . The method of  claim 23 , wherein said glycoprotein-conjugate comprises the formula (121) or (122): 
       
         
           
           
               
               
           
         
         wherein: 
         Pr, L, D, r, x, y, b, d, e, p, Q, Su, and G are as defined in claim  1 . 
       
     
     
         27 . The method of  claim 23 , wherein said glycoprotein-conjugate comprises the formula (123) or (124): 
       
         
           
           
               
               
           
         
         wherein: 
         Pr, L, D, r, x, y, b, d, e, p, Q, Su, G, R 9  are as defined in claim  1 . 
       
     
     
         28 . The method of  claim 23 , wherein said glycoprotein comprises the formula (125) or (126): 
       
         
           
           
               
               
           
         
         wherein: 
         Pr, L, D, r, x, y, b, d, e, p, Q, Su, G, and R 9  are as defined in claim  1 . 
       
     
     
         29 . The method of  claim 23 , wherein said glycoprotein-conjugate is an antibody-conjugate. 
     
     
         30 . The method of  claim 29 , wherein said antibody-conjugate comprises monoclonal antibody. 
     
     
         31 . The method of  claim 29 , wherein D is a pharmaceutically active substance. 
     
     
         32 . The method of  claim 31 , wherein D comprises a cytotoxin, antiviral agent, antibacterial agent, peptide, or oligonucleotide. 
     
     
         33 . The method of  claim 32 , wherein the cytotoxin comprises camptothecins, doxorubicin, daunorubicin, taxanes, calicheamycins, duocarmycins, maytansines, auristatins, or pyrrolobenzodiazepines (PBDs). 
     
     
         34 . The method of  claim 32 , wherein the cytotoxin is selected from the group consisting of colchicine,  vinca  alkaloids, camptothecins, doxorubicin, daunorubicin, taxanes, calicheamycins, tubulysins, irinotecans, an inhibitory peptide, amanitin, deBouganin, duocarmycins, maytansines, auristatins, and pyrrolobenzodiazepines (PBDs). 
     
     
         35 . The method of  claim 32 , wherein the cytotoxin is selected from the group consisting of camptothecins, doxorubicin, daunorubicin, taxanes, calicheamycins, duocarmycins, maytansines, auristatins, and pyrrolobenzodiazepines (PBDs). 
     
     
         36 . The method of  claim 32 , wherein the cytotoxin is selected from the group consisting of colchicine,  vinca  alkaloids, tubulysins, irinotecans, an inhibitory peptide, amanitin and deBouganin. 
     
     
         37 . The method of  claim 23 , wherein the disease comprises cancer. 
     
     
         38 . The method of  claim 37 , wherein the cancer comprises breast cancer. 
     
     
         39 . The method of  claim 37 , wherein the breast cancer comprises HER2-positive breast cancer.