US2023073034A1PendingUtilityA1

Multispecific antibodies, compositions comprising the same, and vectors and uses thereof

66
Assignee: APRILBIO CO LTDPriority: Jan 24, 2020Filed: Jan 23, 2021Published: Mar 9, 2023
Est. expiryJan 24, 2040(~13.5 yrs left)· nominal 20-yr term from priority
Inventors:Sang Hoon Cha
C07K 16/18C07K 16/241C07K 2317/31C07K 16/244C07K 2317/94C07K 16/249C07K 2317/92C07K 2317/56C07K 2317/55C07K 2317/76C07K 2317/622C07K 2317/62C07K 16/2875C07K 16/2866A61K 2039/505
66
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Claims

Abstract

The present disclosure provides multispecific antibodies having increased in vivo sustainability, the multispecific antibodies comprising one or more bioactive effector moieties linked to either or both of an N-terminal and a C-terminal of an antigen binding fragment Fab that binds to human serum albumin.

Claims

exact text as granted — not AI-modified
What is claimed is: 
     
         1 . A multispecific antibody comprising a structural formula of: 
       
         
           
           
               
               
           
         
         wherein the antigen binding fragment (Fab) is a serum albumin Fab; 
         wherein R 1  and R 2  are bioactive effector moieties linked to an N-terminus of the Fab, each linked to a heavy chain variable domain or a light chain variable domain of the Fab; 
         wherein R 3  and R 4  are bioactive effector moieties linked to a C-terminus of the Fab, each linked to a heavy chain variable domain or a light chain variable domain of the Fab; 
         wherein m is 0 or an integer of 1 or greater; and 
         wherein n is 0 or an integer of 1 or greater. 
       
     
     
         2 . The antibody of  claim 1 , wherein R 1  and R 2  are same or different single-chain variable fragments (scFv). 
     
     
         3 . The antibody of  claim 1  or  2 , wherein R 3  and R 4  are same or different Fv fragments or disulfide-stabilized Fv (dsFv) fragments. 
     
     
         4 . The antibody of any of  claims 1 - 3 , wherein each of R 1 , R 2 , R 3 , and R 4  is linked to the Fab by one or more linkers. 
     
     
         5 . The antibody of any of  claims 1 - 4 , wherein the Fab comprises a heavy chain variable domain comprising
 (a) a heavy chain complementarity determining domain 1 (CDR1) comprising the amino acid sequence of SYGIS (SEQ ID NO:61),
 a heavy chain CDR2 comprising the amino acid sequence of WINTYSGGTKYAQKFQG (SEQ ID NO:62), and 
 a heavy chain CDR3 comprising the amino acid sequence of 
   
       
         
           
                 
                 
               
                     
                   (SEQ ID NO: 63) 
                 
                 
                 
                 
               
                     
                     
                   LGHCQRGICSDALDT 
                 
             
                
               
            
             
                
               
            
           
         
         (b) a heavy chain complementarity determining domain 1 (CDR1) comprising the amino acid sequence of SYGIS (SEQ ID NO:61),
 a heavy chain CDR2 comprising the amino acid sequence of RINTYNGNTGYAQRLQG (SEQ ID NO:64), and 
 a heavy chain CDR3 comprising the amino acid sequence of 
 
       
       
         
           
                 
                 
               
                     
                   (SEQ ID NO: 63) 
                 
                 
                 
                 
               
                     
                     
                   LGHCQRGICSDALDT 
                 
             
                
               
            
             
                
               
            
           
         
         (c) a heavy chain complementarity determining domain 1 (CDR1) comprising the amino acid sequence of NYGIH (SEQ ID NO:65),
 a heavy chain CDR2 comprising the amino acid sequence of SISYDGSNKYYADSVKG (SEQ ID NO:66), and 
 a heavy chain CDR3 comprising the amino acid sequence of DVHYYGSGSYYNAFDI (SEQ ID NO:67; 
 
       
       
         
           
                 
                 
               
                     
                   (SEQ ID NO: 67) 
                 
                 
                 
                 
               
                     
                     
                   DVHYYGSGSYYNAFDI 
                 
             
                
               
            
             
                
               
            
           
         
         (d) a heavy chain complementarity determining domain 1 (CDR1) comprising the amino acid sequence of SYAMS (SEQ ID NO:68),
 a heavy chain CDR2 comprising the amino acid sequence of VISHDGGFQYYADSVKG (SEQ ID NO:69), and 
 a heavy chain CDR3 comprising the amino acid sequence of 
 
       
       
         
           
                 
                 
               
                     
                   (SEQ ID NO: 70) 
                 
                 
                 
                 
               
                     
                     
                   AGWLRQYGMDV 
                 
             
                
               
            
             
                
               
            
           
         
         (e) a heavy chain complementarity determining domain 1 (CDR1) comprising the amino acid sequence of AYWIA (SEQ ID NO:71),
 a heavy chain CDR2 comprising the amino acid sequence of MIWPPDADARYSPSFQG (SEQ ID NO:72), and 
 a heavy chain CDR3 comprising the amino acid sequence of LYSGSYSP (SEQ ID NO:73); or 
 
         (f) a heavy chain complementarity determining domain 1 (CDR1) comprising the amino acid sequence of AYSMN (SEQ ID NO:74),
 a heavy chain CDR2 comprising the amino acid sequence of SISSSGRYIHYADSVKG (SEQ ID NO:75), and 
 a heavy chain CDR3 comprising the amino acid sequence of ETVMAGKALDY (SEQ ID NO:76). 
 
       
     
     
         6 . The antibody of any of  claims 1 - 5 , wherein the Fab comprises a light chain variable domain comprising
 (g) a light chain complementarity determining domain 1 (CDR1) comprising the amino acid sequence of RASQSISRYLN (SEQ ID NO:77),
 a light chain CDR2 comprising the amino acid sequence of GASRLES (SEQ ID NO:78), and 
 a light chain CDR3 comprising the amino acid sequence of QQSDSVPVT (SEQ ID NO:79); 
   (h) a light chain complementarity determining domain 1 (CDR1) comprising the amino acid sequence of RASQSISSYLN (SEQ ID NO:80),
 a light chain CDR2 comprising the amino acid sequence of AASSLQS (SEQ ID NO:81), and 
 a light chain CDR3 comprising the amino acid sequence of QQSYSTPPYT (SEQ ID NO:82); 
   (i) a light chain complementarity determining domain 1 (CDR1) comprising the amino acid sequence of RASQSIFNYVA (SEQ ID NO:83),
 a light chain CDR2 comprising the amino acid sequence of DASNRAT (SEQ ID NO:84), and 
 a light chain CDR3 comprising the amino acid sequence of QQRSKWPPTWT (SEQ ID NO:85); 
   (j) a light chain complementarity determining domain 1 (CDR1) comprising the amino acid sequence of RASETVSSRQLA (SEQ ID NO:86),
 a light chain CDR2 comprising the amino acid sequence of GASSRAT (SEQ ID NO:87), and 
 a light chain CDR3 comprising the amino acid sequence of QQYGSSPRT (SEQ ID NO:88); 
   (k) a light chain complementarity determining domain 1 (CDR1) comprising the amino acid sequence of RASQSVSSSSLA (SEQ ID NO:89),
 a light chain CDR2 comprising the amino acid sequence of GASSRAT (SEQ ID NO:87), and 
 a light chain CDR3 comprising the amino acid sequence of QKYSSYPLT (SEQ ID NO:90); or 
   (l) a light chain complementarity determining domain 1 (CDR1) comprising the amino acid sequence of RASQSVGSNLA (SEQ ID NO:91),
 a light chain CDR2 comprising the amino acid sequence of GASTGAT (SEQ ID NO:92), and 
 a light chain CDR3 comprising the amino acid sequence of QQYYSFLAKT (SEQ ID NO:93). 
   
     
     
         7 . The antibody of any of  claims 1 - 6 , wherein the Fab comprises
 a heavy chain complementarity determining domain 1 (CDR1) comprising the amino acid sequence of AYSMN (SEQ ID NO:74), a heavy chain CDR2 comprising the amino acid sequence of SISSSGRYIHYADSVKG (SEQ ID NO:75), and a heavy chain CDR3 comprising the amino acid sequence of ETVMAGKALDY (SEQ ID NO:76), and   a light chain complementarity determining domain 1 (CDR1) comprising the amino acid sequence of RASQSVGSNLA (SEQ ID NO:91), a light chain CDR2 comprising the amino acid sequence of GASTGAT (SEQ ID NO:92), and a light chain CDR3 comprising the amino acid sequence of QQYYSFLAKT (SEQ ID NO:93).   
     
     
         8 . The antibody of any of  claims 1 - 7 , wherein the Fab comprises a heavy chain variable domain comprising an amino acid sequence having at least 80% identity to SEQ ID NO:94, 95, 96, 97, 98, or 99. 
     
     
         9 . The antibody of any of  claims 1 - 8 , wherein the Fab comprises a light chain variable domain comprising an amino acid sequence having at least 80% identity to SEQ ID NO:100, 101, 102, 103, 104, or 105. 
     
     
         10 . The antibody of any of  claims 1 - 9 , wherein the Fab comprises a heavy chain variable domain comprising an amino acid sequence having at least 80% identity to SEQ ID NO:94, 95, 96, 97, 98, or 99, and a light chain variable domain comprising an amino acid sequence having at least 80% identity to SEQ ID NO:100, 101, 102, 103, 104, or 105, respectively. 
     
     
         11 . The antibody of any of  claims 1 - 10 , wherein the Fab comprises a heavy chain domain comprising an amino acid sequence of SEQ ID NO:45 (V H −CH1 domain) and a light chain domain comprising an amino acid sequence of SEQ ID NO:46 (V L −C L  domain). 
     
     
         12 . The antibody of any of  claims 1 - 11 , wherein each linker comprises 1 to 20 amino acids. 
     
     
         13 . The antibody of any of  claims 1 - 12 , wherein each linker comprises an amino acid sequence having at least 90% identity to SEQ ID NO:3 or SEQ ID NO:4. 
     
     
         14 . The antibody of any of  claims 1 - 13 , wherein each linker comprises an amino acid sequence of SEQ ID NO:3 or SEQ ID NO:4. 
     
     
         15 . The antibody of any of  claims 1 - 14 , wherein each of the R 1  and R 2  is an anti-CD40L hu5c8 scFv. 
     
     
         16 . The antibody of any of  claims 1 - 15 , wherein each of the R 1  and R 2  is an anti-CD40L hu5c8 scFv comprising an amino acid sequence having at least 80% identity to SEQ ID NO:47 or SEQ ID NO:48. 
     
     
         17 . The antibody of any of  claims 1 - 16 , wherein each of the R 1  and R 2  is an anti-CD40L hu5c8 scFv comprising an amino acid sequence of SEQ ID NO:47 or SEQ ID NO:48. 
     
     
         18 . The antibody of any of  claims 1 - 17 , wherein each of R 3  and R 4  is one or more bioactive effector moieties selected from the group consisting of anti-TNF-α Fv, anti-TNF-α disulfide-stabilized Fv (dsFv), anti-TL-23 Fv, anti-TL-23 dsFv, anti-IFNAR1, and anti-IFNAR1 dsFv. 
     
     
         19 . The antibody of any of  claims 1 - 18 , wherein each of R 3  and R 4  is one or more bioactive effector moieties comprising an anti-TNF-α Fv comprising a heavy chain amino acid sequence having 80% identity to SEQ ID NO:49 and a light chain amino acid sequence having 80% identity to SEQ ID NO:50, anti-TNF-α disulfide-stabilized Fv (dsFv) comprising a heavy chain amino acid sequence having 80% identity to SEQ ID NO:51 and a light chain amino acid sequence having 80% identity to SEQ ID NO:52, anti-TL-23 Fv comprising a heavy chain amino acid sequence having 80% identity to SEQ ID NO:53 and a light chain amino acid sequence having 80% identity to SEQ ID NO:54, anti-TL-23 dsFv comprising a heavy chain amino acid sequence having 80% identity to SEQ ID NO:55 and a light chain amino acid sequence having 80% identity to SEQ ID NO:56, anti-IFNAR1 comprising a heavy chain amino acid sequence having 80% identity to SEQ ID NO:57 and a light chain amino acid sequence having 80% identity to SEQ ID NO:58, and/or anti-IFNAR1 dsFv comprising a heavy chain amino acid sequence having 80% identity to SEQ ID NO:59 and a light chain amino acid sequence having 80% identity to SEQ ID NO:60. 
     
     
         20 . The antibody of any of  claims 1 - 19 , wherein each of R 3  and R 4  is one or more bioactive effector moieties comprising an anti-TNF-α Fv comprising a heavy chain of SEQ ID NO:49 and a light chain of SEQ ID NO:50, anti-TNF-α disulfide-stabilized Fv (dsFv) comprising a heavy chain of SEQ ID NO:51 and a light chain of SEQ ID NO:52, anti-IL-23 Fv comprising a heavy chain of SEQ ID NO:53 and a light chain of SEQ ID NO:54, anti-IL-23 dsFv comprising a heavy chain of SEQ ID NO:55 and a light chain of SEQ ID NO:56, anti-IFNAR1 comprising a heavy chain of SEQ ID NO:57 and a light chain of SEQ ID NO:58, and/or anti-IFNAR1 dsFv comprising a heavy chain of SEQ ID NO:59 and a light chain of SEQ ID NO:60. 
     
     
         21 . A composition comprising the multispecific antibody of any of  claims 1 - 20  and an excipient. 
     
     
         22 . A pharmaceutical composition comprising the multispecific antibody of any of  claims 1 - 20  and a pharmaceutically accepted excipient. 
     
     
         23 . A method of treating an autoimmune disease in a subject in need thereof, the method comprising administering the pharmaceutical composition of  claim 22  to the subject. 
     
     
         24 . An expression vector comprising:
 (a) a promoter,   (b) a first nucleic acid molecule encoding an antigen binding fragment (Fab) that binds to serum albumin, and   (c) a second nucleic acid molecule encoding a bioactive effector moiety and a linker,   wherein the promoter, the first nucleic acid sequence, and the second nucleic acid molecules are operably linked.   
     
     
         25 . The vector of  claim 24 , wherein the second nucleic acid molecule encodes 2 or more bioactive effector moieties and linkers. 
     
     
         26 . The vector of  claim 24  or  25 , wherein the first nucleic acid molecule comprises a nucleic acid sequence encoding a Fab comprising a heavy chain variable domain comprising
 (a) a heavy chain complementarity determining domain 1 (CDR1) comprising the amino acid sequence of SYGIS (SEQ ID NO:61),
 a heavy chain CDR2 comprising the amino acid sequence of WINTYSGGTKYAQKFQG (SEQ ID NO:62), and 
 a heavy chain CDR3 comprising the amino acid sequence of LGHCQRGICSDALDT (SEQ ID NO:63; 
 
 
       
         
           
                 
                 
               
                     
                   (SEQ ID NO: 63) 
                 
                 
                 
                 
               
                     
                     
                   LGHCQRGICSDALDT 
                 
             
                
               
            
             
                
               
            
           
         
         (b) a heavy chain complementarity determining domain 1 (CDR1) comprising the amino acid sequence of SYGIS (SEQ ID NO:61),
 a heavy chain CDR2 comprising the amino acid sequence of RINTYNGNTGYAQRLQG (SEQ ID NO:64), and 
 a heavy chain CDR3 comprising the amino acid sequence of LGHCQRGICSDALDT (SEQ ID NO:63; 
 
       
       
         
           
                 
                 
               
                     
                   (SEQ ID NO: 63) 
                 
                 
                 
                 
               
                     
                     
                   LGHCQRGICSDALDT 
                 
             
                
               
            
             
                
               
            
           
         
         (c) a heavy chain complementarity determining domain 1 (CDR1) comprising the amino acid sequence of NYGIH (SEQ ID NO:65),
 a heavy chain CDR2 comprising the amino acid sequence of SISYDGSNKYYADSVKG (SEQ ID NO:66), and 
 a heavy chain CDR3 comprising the amino acid sequence of DVHYYGSGSYYNAFDI (SEQ ID NO:67; 
 
       
       
         
           
                 
                 
               
                     
                   (SEQ ID NO: 67) 
                 
                 
                 
                 
               
                     
                     
                   DVHYYGSGSYYNAFDI 
                 
             
                
               
            
             
                
               
            
           
         
         (d) a heavy chain complementarity determining domain 1 (CDR1) comprising the amino acid sequence of SYAMS (SEQ ID NO:68),
 a heavy chain CDR2 comprising the amino acid sequence of VISHDGGFQYYADSVKG (SEQ ID NO:69), and 
 a heavy chain CDR3 comprising the amino acid sequence of 
 
       
       
         
           
                 
                 
               
                     
                   (SEQ ID NO: 70) 
                 
                 
                 
                 
               
                     
                     
                   AGWLRQYGMDV 
                 
             
                
               
            
             
                
               
            
           
         
         (e) a heavy chain complementarity determining domain 1 (CDR1) comprising the amino acid sequence of AYWIA (SEQ ID NO:71),
 a heavy chain CDR2 comprising the amino acid sequence of MIWPPDADARYSPSFQG (SEQ ID NO:72), and 
 a heavy chain CDR3 comprising the amino acid sequence of LYSGSYSP (SEQ ID NO:73); or 
 
         (f) a heavy chain complementarity determining domain 1 (CDR1) comprising the amino acid sequence of AYSMN (SEQ ID NO:74),
 a heavy chain CDR2 comprising the amino acid sequence of SISSSGRYIHYADSVKG (SEQ ID NO:75), and 
 a heavy chain CDR3 comprising the amino acid sequence of ETVMAGKALDY (SEQ ID NO:76). 
 
       
     
     
         27 . The vector of any of  claims 24 - 26 , wherein the first nucleic acid molecule comprises a nucleic acid sequence encoding a Fab comprising a light chain variable domain comprising
 (g) a light chain complementarity determining domain 1 (CDR1) comprising the amino acid sequence of RASQSISRYLN (SEQ ID NO:77),
 a light chain CDR2 comprising the amino acid sequence of GASRLES (SEQ ID NO:78), and 
 a light chain CDR3 comprising the amino acid sequence of QQSDSVPVT (SEQ ID NO:79); 
   (h) a light chain complementarity determining domain 1 (CDR1) comprising the amino acid sequence of RASQSISSYLN (SEQ ID NO:80),
 a light chain CDR2 comprising the amino acid sequence of AASSLQS (SEQ ID NO:81), and 
 a light chain CDR3 comprising the amino acid sequence of QQSYSTPPYT (SEQ ID NO:82); 
   (i) a light chain complementarity determining domain 1 (CDR1) comprising the amino acid sequence of RASQSIFNYVA (SEQ ID NO:83),
 a light chain CDR2 comprising the amino acid sequence of DASNRAT (SEQ ID NO:84), and 
 a light chain CDR3 comprising the amino acid sequence of QQRSKWPPTWT (SEQ ID NO:85); 
   (j) a light chain complementarity determining domain 1 (CDR1) comprising the amino acid sequence of RASETVSSRQLA (SEQ ID NO:86),
 a light chain CDR2 comprising the amino acid sequence of GASSRAT (SEQ ID NO:87), and 
 a light chain CDR3 comprising the amino acid sequence of QQYGSSPRT (SEQ ID NO:88); 
   (k) a light chain complementarity determining domain 1 (CDR1) comprising the amino acid sequence of RASQSVSSSSLA (SEQ ID NO:89),
 a light chain CDR2 comprising the amino acid sequence of GASSRAT (SEQ ID NO:87), and 
 a light chain CDR3 comprising the amino acid sequence of QKYSSYPLT (SEQ ID NO:90); or 
   (l) a light chain complementarity determining domain 1 (CDR1) comprising the amino acid sequence of RASQSVGSNLA (SEQ ID NO:91),
 a light chain CDR2 comprising the amino acid sequence of GASTGAT (SEQ ID NO:92), and 
 a light chain CDR3 comprising the amino acid sequence of QQYYSFLAKT (SEQ ID NO:93). 
   
     
     
         28 . The vector of any of  claims 24 - 27 , wherein the first nucleic acid molecule comprises a nucleic acid sequence encoding a Fab comprising
 a heavy chain complementarity determining domain 1 (CDR1) comprising the amino acid sequence of AYSMN (SEQ ID NO:74), a heavy chain CDR2 comprising the amino acid sequence of SISSSGRYIHYADSVKG (SEQ ID NO:75), and a heavy chain CDR3 comprising the amino acid sequence of ETVMAGKALDY (SEQ ID NO:76), and   a light chain complementarity determining domain 1 (CDR1) comprising the amino acid sequence of RASQSVGSNLA (SEQ ID NO:91), a light chain CDR2 comprising the amino acid sequence of GASTGAT (SEQ ID NO:92), and a light chain CDR3 comprising the amino acid sequence of QQYYSFLAKT (SEQ ID NO:93).   
     
     
         29 . The vector of any of  claims 24 - 28 , wherein the first nucleic acid molecule comprises a nucleic acid sequence encoding a Fab comprising a heavy chain variable domain comprising an amino acid sequence having at least 80% identity to SEQ ID NO:94, 95, 96, 97, 98, or 99. 
     
     
         30 . The vector of any of  claims 24 - 29 , wherein the first nucleic acid molecule comprises a nucleic acid sequence encoding a Fab comprising a light chain variable domain comprising an amino acid sequence having at least 80% identity to SEQ ID NO:100, 101, 102, 103, 104, or 105. 
     
     
         31 . The vector of any of  claims 24 - 30 , wherein the first nucleic acid molecule comprises a nucleic acid sequence encoding a Fab comprising a heavy chain variable domain comprising an amino acid sequence having at least 80% identity to SEQ ID NO:94, 95, 96, 97, 98, or 99, and a light chain variable domain comprising an amino acid sequence having at least 80% identity to SEQ ID NO:100, 101, 102, 103, 104, or 105, respectively. 
     
     
         32 . The vector of any of  claims 24 - 31 , wherein the first nucleic acid molecule comprises a nucleic acid sequence encoding a Fab comprising a heavy chain domain comprising an amino acid sequence of SEQ ID NO:45 (V H −C H1  domain) and a light chain domain comprising an amino acid sequence of SEQ ID NO:46 (V L −C L  domain). 
     
     
         33 . The vector of any of  claims 24 - 32 , wherein the bioactive effector moieties are anti-TNF-α Fv, anti-TNF-α dsFv, anti-IL-23 Fv, anti-IL-23 dsFv, anti-IFNAR1 Fv, and/or anti-IFNAR1 dsFv. 
     
     
         34 . The vector of any of  claims 23 - 33 , wherein the second nucleic acid molecule comprises a nucleotide sequence encoding the amino acid sequence of one or more of SEQ ID NOs: 49-60.

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