Composition for Treating HBV Infection

Abstract

The present invention provides a composition comprising hepatitis B virus (HBV) component(s), and which may be either nucleic acid- or polypeptide-based as well as nucleic acid molecules and vectors encoding such HBV component(s). It also relates to infectious viral particles and host cells comprising such nucleic acid molecules or vectors. It also provides composition and kits of parts comprising such nucleic acid molecules, vectors, infectious viral particles or host cells and the therapeutic use thereof for preventing or treating HBV infections.

1 .- 59 . (canceled) 
     
     
         60 . An immunogenic composition comprising a combination of at least two polypeptides or of the nucleic acid molecules encoding said at least two polypeptides wherein said at least two polypeptides are selected from the group consisting of:
 (i) A polymerase moiety comprising at least 450 amino acid residues of a polymerase protein originating from a first HBV virus;   (ii) A core moiety comprising at least 100 amino acid residues of a core protein originating from a second HBV virus; and   (iii) An env moiety comprising one or more immunogenic domain(s) of 15 to 100 consecutive amino acid residues of a HBsAg protein originating from a third HBV virus.   
     
     
         61 . The immunogenic composition according to  claim 60 , wherein said composition is selected from the group consisting of (i) a composition comprising a combination of said polymerase moiety and said core moiety or a combination of nucleic acid molecules encoding said polymerase moiety and said core moiety; (ii) a composition comprising a combination of said core moiety and said env moiety or a combination of nucleic acid molecules encoding said core moiety and said env moiety; and (iii) a composition comprising a combination of said polymerase moiety, said core moiety and said env moiety or a combination of nucleic acid molecules encoding said polymerase moiety, said core moiety and said env moiety. 
     
     
         62 . The immunogenic composition according to  claim 60  or  claim 61 , wherein said first, second, and third HBV virus are from different genotypes. 
     
     
         63 . The immunogenic composition according to  claim 60  or  claim 61 , wherein at least two of said first, second, and third HBV viruses are from the same HBV genotype and preferably said first, second, and third HBV viruses are from the same genotype. 
     
     
         64 . The immunogenic composition according to  claim 63 , wherein said first, second, and third HBV viruses are from genotype D and preferably from HBV isolate Y07587. 
     
     
         65 . The immunogenic composition according to  claim 63 , wherein said first, second, and third HBV viruses are from genotype B or C. 
     
     
         66 . The immunogenic composition according to  claim 60 , wherein said polymerase moiety is modified by truncation of at least 20 amino acid residues and at most 335 amino acid residues normally present at the N-terminus of a native HBV polymerase and preferably of the first 47 or 46 amino acid residues following the initiator Met residue located at the N-terminus of a native HBV polymerase. 
     
     
         67 . The immunogenic composition according to  claim 66 , wherein said polymerase moiety comprises an amino acid sequence which exhibits at least 80% of identity with the amino acid sequence shown in SEQ ID NO:7. 
     
     
         68 . The immunogenic composition according to  claim 60 , wherein said polymerase moiety is modified so as to exhibit a reduced reverse-transcriptase (RTase) enzymatic activity with respect to a native HBV polymerase, and preferably comprises the substitution of the Asp residue corresponding to position 540 in SEQ ID NO:1 and to position 494 in SEQ ID NO:7 to any amino acid residue other than Asp, and preferably to a His residue. 
     
     
         69 . The immunogenic composition according to  claim 60 , wherein said polymerase moiety is modified so as to exhibit a reduced RNase H enzymatic activity with respect to a native HBV polymerase, and preferably comprises the substitution of the Glu residue corresponding to position 718 in SEQ ID NO:1 and to position 672 in SEQ ID NO:7 to any amino acid residue other than Glu, and preferably to a His residue. 
     
     
         70 . The immunogenic composition according to  claim 69 , wherein said polymerase moiety comprises an amino acid sequence which exhibits at least 80% of identity with the amino acid sequence shown in SEQ ID NO:8 or with the amino acid sequence shown in SEQ ID NO:7 with the substitution of the Asp residue in position 494 to an His residue and the substitution of the Glu residue in position 672 to an His residue. 
     
     
         71 . The immunogenic composition according to  claim 60 , wherein said polymerase moiety is fused in frame to a heterologous hydrophobic sequence, and preferably in frame to the signal and trans-membrane peptides of the rabies glycoprotein, wherein said rabies signal sequence is fused in frame at the N-terminus and said rabies transmembrane sequence is fused in frame at the C-terminus of said polymerase moiety. 
     
     
         72 . The immunogenic composition according to  claim 71 , wherein said polymerase moiety comprises an amino acid sequence which exhibits at least 80% of identity with the amino acid sequence shown in SEQ ID NO:9. 
     
     
         73 . The immunogenic composition according to  claim 60 , wherein said core moiety is modified by truncation of at least 10 amino acid residues and at most 40 amino acid residues normally present at the C-terminus of a native HBV core, and preferably of the last 35 amino acid residues of a native HBV core. 
     
     
         74 . The immunogenic composition according to  claim 73 , wherein said core moiety comprises an amino acid sequence which exhibits at least 80% of identity with the amino acid sequence shown in SEQ ID NO:10. 
     
     
         75 . The immunogenic composition according to  claim 60 , wherein said core moiety is modified so as to exhibit a reduced recognition of and/or interaction with an HBV envelope protein with respect to a native HBV core, and preferably comprises an amino acid sequence which exhibits at least 80% of identity with the amino acid sequence shown in SEQ ID NO:11. 
     
     
         76 . The immunogenic composition according to  claim 60 , wherein said one or more immunogenic domain(s) in use in the present invention are selected from the group consisting of:
 The portion of an env protein (HBsAg) extending from position 14-51 (env 1 domain);   The portion of an env protein (HBsAg) extending from position 165-194 (env 2 domain);   The portion of an env protein (HBsAg) extending from position 81-106 (env 3 domain);   The portion of an env protein (HBsAg) extending from position 202-226 (env 4 domain); and   Any combination thereof.   
     
     
         77 . The immunogenic composition according to  claim 76 , wherein said immunogenic domain(s) comprises an amino acid sequence which exhibits at least 80% of identity with any of the amino acid sequence shown in SEQ ID NOs 12-15. 
     
     
         78 . The immunogenic composition according to  claim 77 , wherein said env moiety comprises an amino acid sequence which exhibits at least 80% of identity with the amino acid sequence shown in SEQ ID NO:16 or in SEQ ID NO:17. 
     
     
         79 . The immunogenic composition according to  claim 60 , wherein said polymerase, core, and/or env moieties are fused in frame in a single polypeptide chain by pairs or all together. 
     
     
         80 . The immunogenic composition according to  claim 79 , wherein said env moiety is fused in frame to the C-terminus of said core moiety. 
     
     
         81 . The immunogenic composition according to  claim 80 , wherein said fusion polypeptide of the core moiety with the env moiety is selected from the group consisting of:
 A polypeptide comprising an amino acid sequence which exhibits at least 80% of identity with the amino acid sequence shown in SEQ ID NO:18;   A polypeptide comprising an amino acid sequence which exhibits at least 80% of identity with the amino acid sequence shown in SEQ ID NO:19; and   A polypeptide comprising an amino acid sequence which exhibits at least 80% of identity with the amino acid sequence shown in SEQ ID NO:20 or with the portion of the amino acid sequence shown in SEQ ID NO:20 starting at residue 1 and ending at residue 251 or alternatively with the portion of the amino acid sequence shown in SEQ ID NO:20 starting at residue 1 and ending at residue 221; or deleted versions thereof lacking residues 77-84 in SEQ ID NO:20.   
     
     
         82 . A nucleic acid molecule encoding the polymerase moiety as defined in  claim 60  or  claim 66 , the core moiety as defined in  claim 60 ,  claim 73 , or  claim 75 , and the env moiety as defined in  claim 60 ,  claim 76 , or  claim 79 , or any combination thereof. 
     
     
         83 . The nucleic acid molecule according to  claim 82 , wherein said nucleic acid molecule is selected from the group consisting of:
 A nucleic acid molecule comprising a nucleotide sequence which exhibits at least 80% of identity with the nucleotide sequence shown in SEQ ID NO:21;   A nucleic acid molecule comprising a nucleotide sequence which exhibits at least 80% of identity with the nucleotide sequence shown in SEQ ID NO:22;   A nucleic acid molecule comprising a nucleotide sequence which exhibits at least 80% of identity with the nucleotide sequence shown in SEQ ID NO:21 with the substitution of the G nucleotide in position 1480 to a C, the G nucleotide in position 2014 to a C and the A nucleotide in position 2016 to a T;   A nucleic acid molecule comprising a nucleotide sequence which exhibits at least 80% of identity with the nucleotide sequence shown in SEQ ID NO:23;   A nucleic acid molecule comprising a nucleotide sequence which exhibits at least 80% of identity with the nucleotide sequence shown in SEQ ID NO:24;   A nucleic acid molecule comprising a nucleotide sequence which exhibits at least 80% of identity with the nucleotide sequence shown in SEQ ID NO:25; and   A nucleic acid molecule comprising a nucleotide sequence which exhibits at least 80% of identity with the nucleotide sequence shown in SEQ ID NO:26 or with the portion of the nucleotide sequence shown in SEQ ID NO:26 starting at nucleotide 1 and ending at nucleotide 753 or with the portion of the nucleotide sequence shown in SEQ ID NO:26 starting at nucleotide 1 and ending at nucleotide 663 or deleted versions thereof lacking the portion extending from the G in position 229 to the A in position 252 of SEQ ID NO:26.   
     
     
         84 . A plasmid or viral vector comprising one or more nucleic acid molecule(s) defined in  claim 82  or  claim 83 . 
     
     
         85 . The vector according to  claim 84 , wherein said viral vector is a replication-defective adenoviral vector, and preferably an adenoviral vector comprising said nucleic acid molecule(s) inserted in replacement of the E1 region. 
     
     
         86 . The vector according to  claim 84 , wherein said viral vector is a poxviral vector and more particularly obtained from canarypox, fowlpox, or vaccinia virus, said vaccinia virus being preferably from the modified Ankara (MVA) strain. 
     
     
         87 . The vector according to  claim 84 ,  claim 85 , or  claim 86 , wherein said vector carries the nucleic acid molecules encoding said polymerase moiety, said core moiety and said env moiety. 
     
     
         88 . The vector according to  claim 84 ,  claim 85 , or  claim 86 , wherein said vector carries only one or two of the nucleic acid molecules encoding said polymerase moiety, said core moiety and said env moiety. 
     
     
         89 . The vector according to  claim 88 , wherein said vector is selected from the group consisting of:
 (i) A MVA vector comprising a nucleic acid molecule placed under the control of a vaccinia promoter such as the 7.5K promoter and encoding a polymerase moiety comprising an amino acid sequence as shown in SEQ ID NO:7, SEQ ID NO:8, or SEQ ID NO:9 or in SEQ ID NO:7 with the substitution of the Asp residue in position 494 to an His residue and the substitution of the Glu residue in position 672 to an His residue;   (ii) A MVA vector comprising a nucleic acid molecule placed under the control of a vaccinia promoter such as the pH5r promoter and encoding a core moiety and an env moiety comprising an amino acid sequence as shown in SEQ ID NO:18 or SEQ ID NO:19;   (iii) An E1-defective Ad vector comprising inserted in place of the E1 region a nucleic acid molecule placed under the control of the CMV promoter and encoding a polymerase moiety comprising an amino acid sequence as shown in SEQ ID NO:7, SEQ ID NO:8, or SEQ ID NO:9 or in SEQ ID NO:7 with the substitution of the Asp residue in position 494 to an His residue and the substitution of the Glu residue in position 672 to an His residue; and   (iv) An E1-defective Ad vector comprising inserted in place of the E1 region a nucleic acid molecule placed under the control of the CMV promoter and encoding a core moiety and an env moiety comprising an amino acid sequence as shown in SEQ ID NO:18, SEQ ID NO:19, or SEQ ID NO:20 or the portion of SEQ ID NO:20 starting at residue 1 and ending at residue 251 or the portion of SEQ ID NO:20 starting at residue 1 and ending at residue 221.   
     
     
         90 . An infectious viral particle comprising the nucleic acid molecule according to  claim 82  or  claim 83  or the vector according to  claim 84 ,  claim 85 ,  claim 86 ,  claim 87 ,  claim 88 , or  claim 89 . 
     
     
         91 . The infectious viral particle according to  claim 90 , wherein said infectious viral particles is produced by a process comprising the steps of:
 (a) introducing the viral vector of the invention into a suitable cell line,   (b) culturing said cell line under suitable conditions so as to allow the production of said infectious viral particle,   (c) recovering the produced infectious viral particle from the culture of said cell line, and   (d) optionally purifying said recovered infectious viral particle.   
     
     
         92 . A host cell comprising the nucleic acid molecule according to  claim 82 , the vector according to  claim 84 , or the infectious viral particle according to  claim 90 . 
     
     
         93 . A composition comprising the nucleic acid molecule according to  claim 82 , the vector according to  claim 84 , or the infectious viral particle according to  claim 90 , or the host cell according to  claim 92  and a pharmaceutically acceptable vehicle. 
     
     
         94 . The composition according to  claim 93  which further comprises one or more adjuvant(s) suitable for systemic or mucosal application in humans. 
     
     
         95 . The composition according to  claim 93  or  claim 94  which is formulated for intramuscular or subcutaneous administration. 
     
     
         96 . The composition according to  claim 93 ,  claim 94 , or  claim 95 , which comprises from about 10 5  to about 10 13  infection units of a viral vector or of an infectious viral particle. 
     
     
         97 . A method for the treatment or prevention of an HBV infection, or an HBV-associated disease or pathologic condition, and preferably for the treatment of a chronic hepatitis B viral infection, said method comprising administering to a human or animal organism in need thereof a therapeutically effective amount of:
 at least one of the HBV moieties as defined in  claim 60 ;   the immunogenic composition according to  claim 60 ;   the nucleic acid molecule according to  claim 82  or  claim 83 ;   the vector according to  claim 84 ;   the infectious viral particle according to  claim 90 ;   the host cell according to  claim 92 ; or   the composition according to  claim 93 .   
     
     
         98 . The method according to  claim 97 , wherein said method is used in combination with a standard of care, and preferably a standard of care comprising cytokines and/or nucleotide or nucleoside analogs selected from the group consisting of lamivudine, entecavir, telbivudine, adefovir, dipivoxil, and tenofovir. 
     
     
         99 . A method of inducing or stimulating an immune response, and preferably a CD4+ and/or CD8+-mediated cellular response, against HBV in a host organism comprising administering to said organism at least one of the HBV moieties as defined in  claim 60 , of the immunogenic composition according to  claim 60 , of the nucleic acid molecule according to  claim 82  or  claim 83 , of the vector according to  claim 84 , of the infectious viral particle according to  claim 90 , of the host cell according to  claim 92 , or of the composition according to  claim 93 , so as to induce or stimulate said immune response. 
     
     
         100 . A kit of parts for use in the treatment of an HBV infection, wherein said kit comprises a plurality of active agents selected from the group consisting of the HBV moieties as defined in  claim 60 , of the immunogenic composition according to  claim 60 , of the nucleic acid molecule according to  claim 82  or  claim 83 , of the vector according to  claim 84 , of the infectious viral particle according to  claim 90 , of the host cell according to  claim 92 , or of the composition according to  claim 93 . 
     
     
         101 . The kit of parts according to  claim 100 , which comprises a first vector comprising a nucleic acid molecule encoding the polymerase moiety as defined in  claim 60  and a second vector comprising a nucleic acid molecule encoding the core moiety and/or the env moiety as defined in  claim 60 . 
     
     
         102 . The kit of parts according to  claim 101 , wherein said first vector is a MVA vector as defined in claim  89 (i) and said second vector is a MVA vector as defined in claim  89 (ii). 
     
     
         103 . The kit of parts according to  claim 101 , wherein said first vector is an Ad vector as defined in claim  89 (iii) and said second vector is an Ad vector as defined in claim  89 (iv). 
     
     
         104 . The kit of parts according to  claim 101 , further comprising a third vector expressing an immunomodulator.