Anti-glypican-3 antibody having improved kinetics in plasma

Abstract

A method of modulating the plasma half-life of anti-glypican 3 antibody, a pharmaceutical composition comprising as an active ingredient the anti-glypican 3 antibody that has a plasma half-life that has been modulated, a method of preparing the anti-glypican 3 antibody and a pharmaceutical composition comprising the anti-glypican 3 antibody as an active ingredient are provided. Disclosed is a method of modulating the plasma half-life of anti-glypican 3 antibody by modifying an amino acid residue that is exposed on the surface of the anti-glypican 3 antibody; and anti-glypican 3 antibody that has a plasma half-life that has been modulated by amino acid residue modification, a pharmaceutical composition comprising as an active ingredient the anti-glypican 3 antibody, and a method of preparing the anti-glypican 3 antibody and producing a pharmaceutical composition comprising the anti-glypican 3 antibody as an active ingredient.

1 - 14 . (canceled) 
     
     
         15 . A method for preparing an antibody with modulated plasma kinetics, said method comprising the steps of:
 (a) culturing a host cell bearing a nucleic acid that encodes the antibody under conditions allowing for expression of the nucleic acid, wherein the antibody has an amino acid sequence altered to causes a modification in the charge of at least one amino acid residue in the constant region in the antibody other than the FcRn binding region; and   (b) recovering the antibody from the host cell culture.   
     
     
         16 . The method according to  claim 15 , wherein the modulation of the plasma kinetics is increase or decrease in a parameter selected from the plasma half life, the mean plasma residence time, and the plasma clearance. 
     
     
         17 . The method according to  claim 15 , wherein the modification in the charge of the amino acid residue is achieved by amino acid substitution. 
     
     
         18 . The method according to  claim 17 , wherein the antibody is an IgG antibody. 
     
     
         19 . The method according to  claim 18 , wherein the antibody is an IgG1 antibody. 
     
     
         20 . The method according to  claim 17 , wherein the modification in the charge of the amino acid residue is achieved by substitution of at least one amino acid residue of an IgG1 antibody with a corresponding amino acid residue of an IgG4 antibody. 
     
     
         21 . The method according to  claim 15 , wherein the FcRn binding region comprises the amino acid residues of the EU numbers 250, 253, 310, 311, 314, 428, 435, and 436 according to the Kabat numbering. 
     
     
         22 . The method according to  claim 20 , wherein the modification in the charge of the amino acid residue is achieved by at least one substitution in the heavy chain constant region shown in SEQ ID NO: 31 selected from:
 (a) substitution of H that is the 151st amino acid residue with Q,   (b) substitution of K that is the 157th amino acid residue with Q,   (c) substitution of R that is the 238th amino acid residue with Q,   (d) substitution of D that is the 239th amino acid residue with E,   (e) substitution of L that is the 241st amino acid residue with M, and   (f) substitution of Q that is the 302nd amino acid residue with E.   
     
     
         23 . The method according to  claim 15 , wherein the antibody is an anti-glypican 3 antibody. 
     
     
         24 . A method of stabilizing an anti-glypican 3 antibody that comprises a complementarity-determining region (CDR) derived from a non-human animal, a framework region (FR) derived from human, and a constant region derived from human, said method comprising the steps of:
 (a) culturing a host cell bearing a nucleic acid that encodes the anti-glypican 3 antibody under conditions allowing for expression of the nucleic acid, wherein the anti-glypican 3 antibody has an amino acid sequence altered to increase in the Tm value of the antibody by a modification of at least one amino acid residue; and   (b) recovering the antibody from the host cell culture.   
     
     
         25 . The method according to  claim 24 , wherein the amino acid residue is present in the FR1 region and/or the FR2 region of the heavy chain or the light chain. 
     
     
         26 . The method according to  claim 25 , wherein an amino acid residue in the FR2 region of the heavy chain is substituted with an amino acid residue of the FR2 region of the VH4 subclass. 
     
     
         27 . The method according to  claim 25 , wherein an amino acid residue in the FR2 region of the light chain is substituted with an amino acid residue of the FR2 region of the VK3 subclass. 
     
     
         28 . The method according to  claim 24 , wherein the substitution of the amino acid residue is achieved by:
 (1) at least one substitution in the heavy chain variable region shown in SEQ ID NO: 1 selected from:   (a) substitution of V that is the 37th amino acid residue with I,   (b) substitution of A that is the 40th amino acid residue with P,   (c) substitution of M that is the 48th amino acid residue with I, and   (d) substitution of L that is the 51st amino acid residue with I;   and/or   (2) at least one substitution in the light chain variable region shown in SEQ ID NO: 7 selected from:   (e) substitution of L that is the 42nd amino acid residue with Q,   (f) substitution of S that is the 48th amino acid residue with A, and   (g) substitution of Q that is the 50th amino acid residue with R.   
     
     
         29 - 71 . (canceled)