Anti-Human Transferrin Receptor Antibody Permeating Blood-Brain Barrier

Abstract

Disclosed are a means to convert compounds having physiological or pharmacological activity and unable to pass through the blood-brain barrier into a form that allows them to pass through the blood-brain barrier, and compounds converted thereby. The means is an anti-human transferrin receptor antibody and the converted compounds are molecular conjugates between physiologically active protein or pharmacologically active low-molecular-weight compounds and an anti-human transferrin receptor antibody.

1 - 69 . (canceled) 
     
     
         70 . A fusion protein comprising the amino acid sequences of a humanized anti-human transferrin receptor antibody and iduronate 2-sulfatase,
 wherein the amino acid sequence of iduronate 2-sulfatase is linked to the light chain of the antibody on the C-terminal side or the N-terminal side of the light chain, or linked to the heavy chain of the antibody on the C-terminal side or the N-terminal side of the heavy chain, and   wherein the light chain variable region and the heavy chain variable region of the antibody are selected from (1) or (2) below:   (1) the light chain variable region comprising the amino acid sequence having an identity not lower than 90% to the amino acid sequence set forth as SEQ ID NO: 191, and comprising the amino acid sequence set forth as SEQ ID NO:16 as CDR1, the amino acid sequence set forth as SEQ ID NO:18 as CDR2, and the amino acid sequence set forth as SEQ ID NO:20 as CDR3, and   the heavy chain variable region comprising the amino acid sequence having an identity not lower than 90% to the amino acid sequence set forth as SEQ ID NO: 205, and comprising the amino acid sequence set forth as SEQ ID NO:88 as CDR1, the amino acid sequence set forth as SEQ ID NO:90 as CDR2, and the amino acid sequence set forth as SEQ ID NO:92 as CDR3;   (2) the light chain variable region comprising the amino acid sequence having an identity not lower than 90% to the amino acid sequence set forth as SEQ ID NO: 191, and comprising the amino acid sequence set forth as SEQ ID NO:16 as CDR1, the amino acid sequence set forth as SEQ ID NO:18 as CDR2, and the amino acid sequence set forth as SEQ ID NO:20 as CDR3, and   the heavy chain variable region comprising the amino acid sequence having an identity not lower than 90% to the amino acid sequence set forth as SEQ ID NO: 205, and comprising the amino acid sequence set forth as SEQ ID NO:89 as CDR1, the amino acid sequence set forth as SEQ ID NO:90 as CDR2, and the amino acid sequence set forth as SEQ ID NO:92 as CDR3.   
     
     
         71 . A fusion protein comprising the amino acid sequences of a humanized anti-human transferrin receptor antibody and iduronate 2-sulfatase, wherein the amino acid sequence of the antibody is selected from the group consisting of (1) to (4) below:
 (1) the light chain variable region of the antibody comprises the amino acid sequence set forth as SEQ ID NO:191, and the heavy chain variable region of the antibody comprises the amino acid sequence set forth as SEQ ID NO:205;   (2) the light chain variable region of the antibody comprises the amino acid sequence set forth as SEQ ID NO:193, and the heavy chain variable region of the antibody comprises the amino acid sequence set forth as SEQ ID NO:205;   (3) the light chain variable region of the antibody comprises the amino acid sequence set forth as SEQ ID NO:194, and the heavy chain variable region of the antibody comprises the amino acid sequence set forth as SEQ ID NO:205; and   (4) the light chain variable region of the antibody comprises the amino acid sequence set forth as SEQ ID NO:195, and the heavy chain variable region of the antibody comprises the amino acid sequence set forth as SEQ ID NO:205.   
     
     
         72 . A fusion protein comprising the amino acid sequences of a humanized anti-human transferrin receptor antibody and iduronate 2-sulfatase, wherein the amino acid sequence of the antibody is selected from the group consisting of (1) to (8) below:
 (1) the light chain of the antibody comprises the amino acid sequence set forth as SEQ ID NO:196, and the heavy chain of the antibody comprises the amino acid sequence set forth as SEQ ID NO:210;   (2) the light chain of the antibody comprises the amino acid sequence set forth as SEQ ID NO:198, and the heavy chain of the antibody comprises the amino acid sequence set forth as SEQ ID NO:210;   (3) the light chain of the antibody comprises the amino acid sequence set forth as SEQ ID NO:200, and the heavy chain of the antibody comprises the amino acid sequence set forth as SEQ ID NO:210;   (4) the light chain of the antibody comprises the amino acid sequence set forth as SEQ ID NO:202, and the heavy chain of the antibody comprises the amino acid sequence set forth as SEQ ID NO:210;   (5) the light chain of the antibody comprises the amino acid sequence set forth as SEQ ID NO:196, and the heavy chain of the antibody comprises the amino acid sequence set forth as SEQ ID NO:212;   (6) the light chain of the antibody comprises the amino acid sequence set forth as SEQ ID NO:198, and the heavy chain of the antibody comprises the amino acid sequence set forth as SEQ ID NO:212;   (7) the light chain of the antibody comprises the amino acid sequence set forth as SEQ ID NO:200, and the heavy chain of the antibody comprises the amino acid sequence set forth as SEQ ID NO:212; and   (8) the light chain of the antibody comprises the amino acid sequence set forth as SEQ ID NO:202, and the heavy chain of the antibody comprises the amino acid sequence set forth as SEQ ID NO:212.   
     
     
         73 . The fusion protein according to  claim 70 , wherein the antibody has an affinity to both the extracellular region of human transferrin receptor and the extracellular region of monkey transferrin receptor. 
     
     
         74 . The fusion protein according to  claim 73 , wherein the dissociation constant of the antibody's complex with the extracellular region of human transferrin receptor is not greater than 1×10 −8  M, and the dissociation constant of the antibody's complex with the extracellular region of monkey transferrin receptor is not greater than 5×10 −8  M. 
     
     
         75 . The fusion protein according to  claim 70 , wherein the antibody is Fab antibody, F(ab′)2 antibody, or F(ab′) antibody. 
     
     
         76 . The fusion protein according to  claim 70 , wherein the antibody is a single-chain antibody selected from the group consisting of scFab, scF(ab′), scF(ab′)2 and scFv. 
     
     
         77 . The fusion protein according to  claim 76 , wherein the light chain and the heavy chain thereof are linked via a linker sequence. 
     
     
         78 . The fusion protein according to  claim 76 , wherein the heavy chain is linked, via a linker sequence, to the light chain on the C-terminal side of the light chain. 
     
     
         79 . The fusion protein according to  claim 76 , wherein the light chain is linked, via a linker sequence, to the heavy chain on the C-terminal side of the heavy chain. 
     
     
         80 . The fusion protein according to  claim 77 , wherein the linker sequence consists of 8 to 50 amino acid residues. 
     
     
         81 . The fusion protein according to  claim 80 , wherein the linker sequence is selected from the group consisting of the amino acid sequence Gly-Ser, the amino acid sequence Gly-Gly-Ser, the amino acid sequence Gly-Gly-Gly, the amino acid sequences set forth as SEQ ID NO:3, SEQ ID NO:4, and SEQ ID NO:5, and the amino acid sequence consisting of three consecutively linked amino acid sequences each set forth as SEQ ID NO:3. 
     
     
         82 . The fusion protein according to  claim 70 , wherein the iduronate 2-sulfatase is linked, via a linker sequence, to the light chain on the C-terminal side or the N-terminal side of the light chain, or to the heavy chain on the C-terminal side or the N-terminal side of the heavy chain. 
     
     
         83 : The fusion protein according to  claim 81 , wherein the linker sequence consists of 1 to 50 amino acid residues. 
     
     
         84 . The fusion protein according to  claim 81 , wherein the linker sequence comprises an amino acid sequence selected from the group consisting of a single glycine, a single serine, the amino acid sequence Gly-Ser, the amino acid sequence Gly-Gly-Ser, the amino acid sequence set forth as SEQ ID NO:3, the amino acid sequence set forth as SEQ ID NO:4, the amino acid sequence set forth as SEQ ID NO:5, and amino acid sequence consisting of 1 to 10 of any of said amino acid sequences that are consecutively linked. 
     
     
         85 . The fusion protein according to  claim 70 , wherein the iduronate 2-sulfatase is a human iduronate 2-sulfatase. 
     
     
         86 . The fusion protein according to  claim 85 , wherein the light chain of the antibody has the amino acid sequence set forth as SEQ ID NO:196, and wherein the heavy chain of the antibody is linked, on the C-terminal side of the heavy chain and via a linker sequence Gly-Ser, to the human iduronate 2-sulfatase, and the whole linked heavy chain has the amino acid sequence set forth as SEQ ID NO:251. 
     
     
         87 . The fusion protein according to  claim 85 , wherein the fusion protein comprises the light chain having the amino acid sequence set forth as SEQ ID NO:196, and the heavy chain having the amino acid sequence set forth as SEQ ID NO:210 which is linked, on the C-terminal side of the heavy chain and via a linker sequence Gly-Ser, to the human iduronate 2-sulfatase set forth as SEQ ID NO:246. 
     
     
         88 . The fusion protein according to  claim 85 , wherein the fusion protein comprises a conjugate composed of the human iduronate 2-sulfatase linked via a linker sequence Gly-Ser to the heavy chain on the C-terminal side of the heavy chain, and the light chain, wherein the amino acid sequence of the conjugate is set forth as SEQ ID NO:251, and the amino acid sequence of the light chain is set forth as SEQ ID NO:196. 
     
     
         89 . A DNA fragment encoding the amino acid sequence of the fusion protein according to  claim 70 . 
     
     
         90 . An expression vector comprising the DNA fragment according to  claim 89  that is incorporated therein. 
     
     
         91 . A mammalian cell transformed with the expression vector according to  claim 90 . 
     
     
         92 . A method of treating a disease condition of the central nervous system accompanying Hunter syndrome in a subject in need thereof comprising parenterally administering the fusion protein of  claim 70  to the subject.