Novel glycan conjugates and methods of use thereof

Abstract

The present disclosure is directed to vaccines, antibodies, and/or immunogenic conjugate compositions targeting the SSEA3/SSEA4/GloboH associated epitopes (natural and modified) which elicit antibodies and/or binding fragment production useful for modulating the globo-series glycosphingolipid synthesis. The present disclosure relates to methods and compositions which can modulate the globo-series glycosphingolipid synthesis. Particularly, the present disclosure is directed to glycoenzyme inhibitor compound and compositions and methods of use thereof that can modulate the synthesis of globo-series glycosphingolipid SSEA3/SSEA4/GloboH in the biosynthetic pathway; particularly, the glycoenzyme inhibitors target the alpha-4GalT; beta-4GalNAcT-I; or beta-3GalT-V enzymes in the globo-series synthetic pathway. Moreover, the present disclosure is also directed to the method of using the compositions described herein for the treatment or detection of hyperproliferative diseases and/or conditions.

1 . An immunogenic composition comprising:
 (a) a glycan conjugate including a carrier and one or more glycans, and optionally (b) an adjuvant;   wherein each of the one or more glycans is conjugated with the carrier through a linker having the formula (III):   
       
         
           
           
               
               
           
         
       
       wherein:
 X 1  is —OR or —SR, wherein R is an oxygen or sulfur protecting group, optionally substituted C 1-10  alkyl, optionally substituted aryl, optionally substituted acyl, or optionally substituted imidoyl; 
 each instance of R 1 , R 2 , R 3 , R 4 , R 5 , R 6  and L is independently selected from hydrogen, halogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heterocyclyl, optionally substituted aryl, —N 3 , —NO 2 , —N(R B ) 2 , —N(R A )C(O)R A , —OR A , —OC(O)R A , —CR A , —C(O)N(R B ) 2 , —CN, —C(O)R A , —C(O)OR A , —S(O)R A , —SO 2 R A , —SO 2 N(R B ) 2 , and —NHSO 2 R B ; 
 each instance of R A  is independently selected from hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heterocyclyl, and optionally substituted aryl; 
 each instance of R B  is independently selected from hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heterocyclyl, and optionally substituted aryl; and 
 provided the glycan conjugate is not of the formula (III-a) and (III-b): 
 
       
         
           
           
               
               
           
         
       
     
     
         2 . The immunogenic composition of  claim 1 , wherein L is —OH. 
     
     
         3 . The immunogenic composition of  claim 2 , wherein at least one instance of R 1 , R 2 , R 3 , R 4 , R 5  and R 6  is —N 3 . 
     
     
         4 . The immunogenic composition of  claim 2 , wherein at least one instance of R 1 , R 2 , R 3 , R 4 , R 5  and R 6  is —F. 
     
     
         5 . The immunogenic composition of  claim 1 , wherein L is of the formula: 
       
         
           
           
               
               
           
         
       
       wherein:
 each instance of R 8 , R 9 , R 10  and R 11  is independently selected from hydrogen, halogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heterocyclyl, optionally substituted aryl, —N 3 , —NO 2 , —N(R B ) 2 , —N(R A )C(O)R A , —OR A , —OC(O)R A , —CR A , —C(O)N(R B ) 2 , —CN, —C(O)R A , —C(O)OR A , —S(O)R A , —SO 2 R A , —SO 2 N(R B ) 2 , and —NHSO 2 R B ; wherein R 12  is H, OH, or halogen; 
 R N  is selected from —N 3 , —NO 2 , —N(R B ) 2 , —N(R A )C(O)R A , —OR A , —OC(O)R A , —SR A , —C(O)N(R B ) 2 , —CN, —C(O)R A , —C(O)OR A , —S(O)R A , —SO 2 R A , —SO 2 N(R B ) 2 , and —NHSO 2 R B ; 
 each instance of R A  is independently selected from hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heterocyclyl, and optionally substituted aryl; 
 each instance of R B  is independently selected from hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heterocyclyl, and optionally substituted aryl. 
 
     
     
         6 . The immunogenic composition of  claim 5 , wherein the glycan conjugate is of formula (IV): 
       
         
           
           
               
               
           
         
       
     
     
         7 . The immunogenic composition of  claim 6 , wherein at least one instance of R 1 , R 2 , R 3 , R 8 , R 9 , R 10 , and R 11  is —N 3 . 
     
     
         8 . The immunogenic composition of  claim 6 , wherein at least one instance of R 2 , R 3 , R 8 , R 9 , R 10 , and R 11  is —F. 
     
     
         9 . The immunogenic composition of  claim 1 , wherein the carrier is a protein, a lipid, a lipolized protein, a virus, a peptide, or a dendrimer of glycopeptides. 
     
     
         10 . The immunogenic composition of  claim 9 , wherein the carrier is a protein selected from the group consisting of tetanus toxoid (TT), diphtheria toxoid (DT), diphtheria toxin cross-reacting material  197  (CRM 197 ), fragment C of TT, Keyhole limpet hemocyanin (KLH), bovine serum albumin (BSA), protein D, outer-membrane protein (OMP) and pneumolysin. 
     
     
         11 . The immunogenic composition of  claim 10 , wherein the carrier protein is selected from the group consisting of TT, DT and CRM 197.    
     
     
         12 . The immunogenic composition of  claim 11 , wherein the glycan conjugate is of the formula (IV-a) or (IV-b): 
       
         
           
           
               
               
           
         
       
       wherein m is an integer of 1 to 40, inclusive. 
     
     
         13 . The immunogenic composition of  claim 1 , wherein the linker is a hetero- or homo-bifunctional linker. 
     
     
         14 . The immunogenic composition of  claim 1 , wherein the adjuvant is a glycolipid capable of binding a CD1d molecule on a dendritic cell. 
     
     
         15 . The immunogenic composition of  claim 1 , wherein the adjuvant is C34, 7DW8-5, C17, C23, Gluco-C34, Aluminum salt, Squalene, MF59, or QS-21. 
     
     
         16 . The immunogenic composition of  claim 1 , wherein the immunogenic composition is capable of eliciting an immune response against a cancer cell. 
     
     
         17 . The immunogenic composition of  claim 16 , wherein the cancer cell is selected from the group consisting of a brain cancer cell, a lung cancer cell, a breast cancer cell, an oral cancer cell, an esophagus cancer cell, a stomach cancer cell, a liver cancer cell, a bile duct cancer cell, a pancreatic cancer cell, a colon cancer cell, a kidney cancer cell, a bone cancer cell, a skin cancer cell, a cervical cancer cell, an ovarian cancer cell, and a prostate cancer cell. 
     
     
         18 . The immunogenic composition of  claim 16 , wherein the immune response includes generation of antibodies that specifically bind to one or more of the antigens selected from the group consisting of SSEA3 and SSEA4. 
     
     
         19 . The immunogenic composition of  claim 18 , wherein the antibodies neutralize one or more of SSEA3 and SSEA4 antigen expressed on the surface of cancer cells or cancer stem cells. 
     
     
         20 . The immunogenic composition of  claim 18 , wherein the antibodies predominantly includes IgG antibodies. 
     
     
         21 . A cancer vaccine, comprising a therapeutically effective amount of the immunogenic composition of  claim 1  and a pharmaceutically acceptable excipient. 
     
     
         22 . The cancer vaccine of  claim 21 , wherein the cancer vaccine is able to induce an anti-cancer immune response in a subject. 
     
     
         23 . A method of treating cancer in a subject in need thereof wherein the method comprising administering a therapeutically effective amount of the immunogenic composition of  claim 1 . 
     
     
         24 . The method of  claim 23  wherein the vaccine is co-administered in combination with another therapeutic agent. 
     
     
         25 . The method of  claim 23 , wherein the cancer is selected from the group consisting of brain cancer, lung cancer, breast cancer, oral cancer, esophageal cancer, stomach cancer, liver cancer, bile duct cancer, pancreatic cancer, colon cancer, kidney cancer, bone cancer, skin cancer, cervical cancer, ovarian cancer, and prostate cancer. 
     
     
         26 . The method of  claim 25 , wherein the cancer cell expresses SSEA3 and/or SSEA4 antigen on the surface of the cell. 
     
     
         27 . The method of  claim 26 , wherein the subject is a human. 
     
     
         28 . The method of  claim 23 , wherein the immunogenic composition or the cancer vaccine is administered subcutaneously. 
     
     
         29 . An isolated monoclonal antibody and/or a binding fragment thereof raised against the immunogenic composition of  claim 1 . 
     
     
         30 . A composition comprising an effective amount of the antibody or antigen-binding fragment of  claim 29  and a pharmaceutically acceptable carrier. 
     
     
         31 . A method for making the immunogenic composition of  claim 1 . 
     
     
         32 . The method of  claim 31  wherein the method comprising:
 providing a carrier; 
 conjugating one or more glycan to the carrier by conjugation reaction; 
 wherein each of the one or more glycan is SSEA3 or SSEA4. 
 
     
     
         33 . An immunogenic composition comprising multivalent construct targeting one or more of SSEA4 and SSEA3 and their analogs thereof wherein the glycans are linked to a template and a carrier, 
       
         
           
           
               
               
           
         
         wherein n can be an integer from 1 to 10; 
         wherein glycan can be selected from the group consisting of Formulas I, II, III, and IV; 
         wherein if n is 2 or more, each glycan can be the same as another glycan on the aspartyl peptide or a difference glycan on the aspartyl peptide. 
       
     
     
         34 . The composition of  claim 33  wherein the glycan is selected from the group consisting of SSEA3 and SSEA4. 
     
     
         35 . The composition of  claim 33  wherein the multivalent construct has the structure: 
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         wherein R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , and L on each glycan moiety can be the same or different. 
       
     
     
         36 . A compound having the formula (I): 
       
         
           
           
               
               
           
         
       
       or a salt thereof, 
       wherein:
 X 1  is —OR or —SR, wherein R is hydrogen, a oxygen or sulfur protecting group, optionally substituted C 1-10  alkyl, optionally substituted aryl, optionally substituted acyl, or optionally substituted imidoyl; 
 each instance of R 1 , R 2 , R 3 , R 4 , R 5 , R 6  and L is independently selected from hydrogen, halogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heterocyclyl, optionally substituted aryl, —N 3 , —NO 2 , —N(R B ) 2 , —N(R A )C(O)R A , —OR A , —OC(O)R A , —SR A , —C(O)N(R B ) 2 , —CN, —C(O)R A , —C(O)OR A , —S(O)R A , —SO 2 R A , —SO 2 N(R B ) 2 , and —NHSO 2 R B ; 
 each instance of R A  is independently selected from hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heterocyclyl, and optionally substituted aryl; 
 each instance of R B  is independently selected from hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heterocyclyl, and optionally substituted aryl; and 
 provided the compound is not of the formula (I-a) or (I-b): 
 
       
         
           
           
               
               
           
         
       
     
     
         37 . The compound of  claim 35 , wherein L is —OH. 
     
     
         38 . The compound of  claim 35 , wherein L is of the formula: 
       
         
           
           
               
               
           
         
       
       wherein:
 each instance of R 8 , R 9 , R 10  and R 11  is independently selected from hydrogen, halogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heterocyclyl, optionally substituted aryl, —N 3 , —NO 2 , —N(R B ) 2 , —N(R A )C(O)R A , —OR A , —OC(O)R A , —SR A , —C(O)N(R B ) 2 , —CN, —C(O)R A , —C(O)OR A , —S(O)R A , —SO 2 R A , —SO 2 N(R B ) 2 , and —NHSO 2 R B ; wherein R 12  is H, OH, or halogen; 
 R N  is selected from —N 3 , —NO 2 , —N(R B ) 2 , —N(R A )C(O)R A , —OR A , —OC(O)R A , —SR A , —C(O)N(R B ) 2 , —CN, —C(O)R A , —C(O)OR A , —S(O)R A , —SO 2 R A , —SO 2 N(R B ) 2 , and —NHSO 2 R B ; 
 each instance of R A  is independently selected from hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heterocyclyl, and optionally substituted aryl; and 
 each instance of R B  is independently selected from hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heterocyclyl, and optionally substituted aryl. 
 
     
     
         39 . The compound of  claim 35 , wherein the compound is of Formula (II): 
       
         
           
           
               
               
           
         
       
     
     
         40 . The compound of  claim 39 , wherein at least one instance of R 1 , R 2 , R 3 , R 8 , R 9 , R 10  and R 11  is —F. 
     
     
         41 . The compound of  claim 39 , wherein at least one instance of R 1 , R 2 , R 3 , R 8 , R 9 , R 10  and R 11  is —N 3 . 
     
     
         42 . The method of treating hyperproliferative disease or condition comprising the administering to a subject in need thereof a therapeutically effective amount of the compound of  claim 36 .