Vaccines against hepatitis b virus

Abstract

The present application provides immunotherapies for Hepatitis B virus infections. Provided herein are genetically modified arenaviral vectors suitable as vaccines for prevention and treatment of Hepatitis B virus infections. Also provided herein are pharmaceutical compositions and methods for the treatment of Hepatitis B virus infections. Specifically, provided herein are pharmaceutical compositions, vaccines, and methods of treating Hepatitis B virus infection.

What is claimed is: 
     
         1 . An infectious arenavirus viral vector, wherein an arenavirus open reading frame is removed and replaced by a nucleotide sequence selected from the group consisting of:
 a. a nucleotide sequence encoding an HBV pre-S2/S protein or an antigenic fragment thereof;   b. a nucleotide sequence encoding an HBV HBc protein or an antigenic fragment thereof;   c. a nucleotide sequence encoding an HBV HBs protein or an antigenic fragment thereof;   d. a nucleotide sequence encoding a fusion of HBV HBs and HBc proteins or antigenic fragments thereof; and   e. a nucleotide sequence encoding an HBV HBe protein or an antigenic fragment thereof.   
     
     
         2 . The viral vector of  claim 1  wherein the pre-S2/S protein or the antigenic fragment thereof comprises an amino acid sequence that is 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence encoded by the nucleotide sequence of SEQ ID NO: 1. 
     
     
         3 . The viral vector of  claim 1  wherein the HBc protein or the antigenic fragment thereof comprises an amino acid sequence that is 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence encoded by the nucleotide sequence of SEQ ID NO: 2. 
     
     
         4 . The viral vector of  claim 1  wherein fusion of HBV HBs and HBc proteins or antigenic fragments thereof comprises an amino acid sequence that is 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence encoded by the nucleotide sequence of SEQ ID NO: 3. 
     
     
         5 . The viral vector of  claim 1  wherein the HBe protein or the antigenic fragment thereof comprises an amino acid sequence that is 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence encoded by the nucleotide sequence of SEQ ID NO: 26. 
     
     
         6 . The viral vector of  claim 1  comprising at least two of:
 a. a nucleotide sequence encoding an HBV pre-S2/S protein or an antigenic fragment thereof; 
 b. a nucleotide sequence encoding an HBV HBc protein or an antigenic fragment thereof; 
 c. a nucleotide sequence encoding an HBV HBs protein or an antigenic fragment thereof; 
 d. a nucleotide sequence encoding a fusion of HBV HBs and HBc proteins or antigenic fragments thereof; and 
 e. a nucleotide sequence encoding an HBV HBe protein or an antigenic fragment thereof. 
 
     
     
         7 . The viral vector of  claim 1  comprising at least three of:
 a. a nucleotide sequence encoding an HBV pre-S2/S protein or an antigenic fragment thereof; 
 b. a nucleotide sequence encoding an HBV HBc protein or an antigenic fragment thereof; 
 c. a nucleotide sequence encoding an HBV HBs protein or an antigenic fragment thereof; 
 d. a nucleotide sequence encoding a fusion of HBV HBs and HBc proteins or antigenic fragments thereof; and 
 e. a nucleotide sequence encoding an HBV HBe protein or an antigenic fragment thereof. 
 
     
     
         8 . The viral vector of  claim 6  or  7  wherein expression of the nucleotide sequences produces an antigenic protein complex that elicits higher titers of neutralizing antibodies than expression of the protein complex components individually. 
     
     
         9 . The viral vector of any one of the preceding claims wherein the arenavirus is lymphocytic choriomeningitis virus. 
     
     
         10 . The viral vector of any one of the preceding claims wherein the open reading frame that encodes the glycoprotein of the arenavirus is deleted or functionally inactivated. 
     
     
         11 . The viral vector of any one of the preceding claims wherein the genomic information encoding the infectious arenavirus viral vector is derived from the lymphocytic choriomeningitis virus Clone 13 strain. 
     
     
         12 . The viral vector of any one of the preceding claims wherein the genomic information encoding the infectious arenavirus viral vector is derived from the lymphocytic choriomeningitis virus MP strain. 
     
     
         13 . The viral vector of any one of the preceding claims wherein the viral vector comprises a genomic segment, wherein the genomic segment comprises a nucleotide sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, at least 99%, or 100% identical to the sequence of nucleotide 1639 to 3315 of SEQ ID NO: 11 or 1640 to 3316 of SEQ ID NO: 12. 
     
     
         14 . The viral vector of any one of the preceding claims wherein the viral vector comprises a genomic segment comprising a nucleotide sequence encoding an expression product whose amino acid sequence is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, at least 99%, or 100% identical to the amino acid sequence encoded by 1639 to 3315 of SEQ ID NO: 11 or 1640 to 3316 of SEQ ID NO: 12. 
     
     
         15 . The viral vector of any one of the preceding claims wherein the arenavirus is Junin virus. 
     
     
         16 . The viral vector of  claim 15 , wherein the genomic information encoding the infectious arenavirus viral vector is derived from the Junin virus Candid #1 strain. 
     
     
         17 . The viral vector of any one of  claims 1  to  16 , wherein the growth or infectivity of the arenavirus is not affected by the heterologous nucleic acid. 
     
     
         18 . A pharmaceutical composition comprising a viral vector of any one of the preceding claims and a pharmaceutically acceptable carrier. 
     
     
         19 . An immunogenic composition comprising a viral vector of any one of  claims 1  to  17  and a pharmaceutically acceptable carrier. 
     
     
         20 . A vaccine comprising a viral vector of any one of  claims 1  to  17  and a pharmaceutically acceptable carrier. 
     
     
         21 . A method of treating or preventing a Hepatitis B virus infection in a patient, wherein said method comprises administering to the patient a viral vector of any one of  claims 1  to  17 , the pharmaceutical composition of  claim 18 , the immunogenic composition of  claim 19 , or the vaccine of  claim 20 . 
     
     
         22 . Use of a viral vector of any one of  claims 1  to  17 , the pharmaceutical composition of  claim 18 , the immunogenic composition of  claim 19 , or the vaccine of  claim 20  for the treatment or prevention of a Hepatitis B virus infection in a patient. 
     
     
         23 . The use of  claim 22 , wherein the viral vector of any one of  claims 1  to  17 , the pharmaceutical composition of  claim 18 , the immunogenic composition of  claim 19 , or the vaccine of  claim 20  is suitable for intramuscular injection. 
     
     
         24 . The use of  claim 22 , wherein the viral vector of any one of  claims 1  to  17 , the pharmaceutical composition of  claim 18 , the immunogenic composition of  claim 19 , or the vaccine of  claim 20  is suitable for intravenous injection. 
     
     
         25 . An isolated nucleic acid, wherein the nucleic acid comprises an arenavirus genomic segment wherein one open reading frame of the genomic segment is deleted or functionally inactivated and wherein the genomic segment comprises one or more of:
 a. a nucleotide sequence encoding an HBV pre-S2/S protein or an antigenic fragment thereof;   b. a nucleotide sequence encoding an HBV HBc protein or an antigenic fragment thereof;   c. a nucleotide sequence encoding an HBV HBs protein or an antigenic fragment thereof;   d. a nucleotide sequence encoding a fusion of HBV HBs and HBc proteins or antigenic fragments thereof; and   e. a nucleotide sequence encoding an HBV HBe protein or an antigenic fragment thereof.   
     
     
         26 . The isolated nucleic acid of  claim 25 , wherein the genomic segment is the short segment, wherein the open reading frame encoding the GP is deleted. 
     
     
         27 . A method for generating an infectious, replication-deficient arenavirus viral vector comprising:
 a. transfecting into a host cell the nucleic acid of  claim 25  or  26 ;   b. maintaining the host cell under conditions suitable for virus formation; and   c. harvesting the infectious, replication-deficient arenavirus viral vector;   
       wherein the host cell expresses the open reading frame that is deleted or functionally inactivated of the genomic segment. 
     
     
         28 . The arenavirus viral vector of  claim 1 , wherein the arenavirus open reading frame is the glycoprotein (GP) open reading frame. 
     
     
         29 . An infectious, replication-deficient arenavirus viral vector engineered to contain a genome with the ability to amplify and express its genetic information in infected cells but unable to produce further infectious progeny particles in normal, not genetically engineered cells, wherein one arenavirus open reading frame is removed and replaced by a nucleotide sequence encoding an HBV antigen or an antigenic fragment thereof, wherein administration of the arenavirus viral vector to a subject induces a long-lasting immune response against the HBV antigen or an antigenic fragment thereof. 
     
     
         30 . The arenavirus viral vector of  claim 29 , wherein the long-lasting immune response induces a detectable antibody titer against an HBV antigen or an antigenic fragment thereof. 
     
     
         31 . The arenavirus viral vector of  claim 29 , wherein the long-lasting immune response induces a detectable antibody titer against the HBV antigen or an antigenic fragment thereof for at least a minimum of 4 weeks. 
     
     
         32 . The arenavirus viral vector of  claim 30  or  31 , wherein the long lasting immune response increases the antibody titer against the HBV antigen or an antigenic fragment thereof by at least 100%, at least 200%, at least 300%, at least 400%, at least 500%, or at least 1000%. 
     
     
         33 . A pharmaceutical composition comprising a first infectious, replication-deficient arenavirus viral vector engineered to contain a genome with the ability to amplify and express its genetic information in infected cells but unable to produce further infectious progeny particles in normal, not genetically engineered cells, wherein one arenavirus open reading frame is removed and replaced by a first nucleotide sequence selected from the group consisting of:
 a. a nucleotide sequence encoding an HBV pre-S2/S protein or an antigenic fragment thereof;   b. a nucleotide sequence encoding an HBV HBc protein or an antigenic fragment thereof;   c. a nucleotide sequence encoding an HBV HBs protein or an antigenic fragment thereof;   d. a nucleotide sequence encoding a fusion of HBV HBs and HBc proteins or antigenic fragments thereof; and   e. a nucleotide sequence encoding an HBV HBe protein or an antigenic fragment thereof;   and a second infectious, replication-deficient arenavirus viral vector engineered to contain a genome with the ability to amplify and express its genetic information in infected cells but unable to produce further infectious progeny particles in normal, not genetically engineered cells, wherein one arenavirus open reading frame is removed and replaced by a second nucleotide sequence selected from the group consisting of:   a. a nucleotide sequence encoding an HBV pre-S2/S protein or an antigenic fragment thereof;   b. a nucleotide sequence encoding an HBV HBc protein or an antigenic fragment thereof;   c. a nucleotide sequence encoding an HBV HBs protein or an antigenic fragment thereof;   d. a nucleotide sequence encoding a fusion of HBV HBs and HBc proteins or antigenic fragments thereof; and   e. a nucleotide sequence encoding an HBV HBe protein or an antigenic fragment thereof.   
     
     
         34 . The pharmaceutical composition of  claim 33 , wherein the first and second nucleic acid sequences are different. 
     
     
         35 . The pharmaceutical composition of  claim 33  or  34 , wherein the first nucleic acid sequence encodes the HBV pre-S2/S protein or fragment thereof, and wherein the second nucleic acid sequence encodes the HBV HBc protein or a fragment thereof. 
     
     
         36 . The pharmaceutical composition of  claim 33  or  34 , wherein the first nucleic acid sequence encodes the HBV pre-S2/S protein or fragment thereof, and wherein the second nucleic acid sequence encodes the fusion of the HBV HBs and HBc proteins or fragments thereof. 
     
     
         37 . The pharmaceutical composition of  claim 33  or  34 , wherein the first nucleic acid sequence encodes the HBV HBc protein or fragment thereof, and wherein the second nucleic acid sequence encodes the fusion of the HBV HBs and HBc proteins or fragments thereof. 
     
     
         38 . The pharmaceutical composition of  claim 33  or  34 , wherein the first nucleic acid sequence encodes the HBV pre-S2/S protein or fragment thereof, and wherein the second nucleic acid sequence encodes the HBV HBe protein or a fragment thereof. 
     
     
         39 . The pharmaceutical composition of  claim 33  or  34 , wherein the first nucleic acid sequence encodes the HBV HBe protein or fragment thereof, and wherein the second nucleic acid sequence encodes the fusion of the HBV HBs and HBc proteins or fragments thereof. 
     
     
         40 . The pharmaceutical composition of  claim 33  or  34 , wherein the first nucleic acid sequence encodes the HBV HBs protein or fragment thereof, and wherein the second nucleic acid sequence encodes the HBV HBe protein or a fragment thereof. 
     
     
         41 . The pharmaceutical composition of  claim 33  or  34 , wherein the first nucleic acid sequence encodes the HBV pre-S2/S protein or fragment thereof, and wherein the second nucleic acid sequence encodes the HBV HBs protein or a fragment thereof. 
     
     
         42 . The pharmaceutical composition of  claim 33  or  34 , wherein the first nucleic acid sequence encodes the HBV HBs protein or fragment thereof, and wherein the second nucleic acid sequence encodes the HBV HBc protein or a fragment thereof. 
     
     
         43 . The pharmaceutical composition of any one of  claims 33  to  42 , wherein the composition is suitable for intramuscular administration. 
     
     
         44 . The pharmaceutical composition of any one of  claims 33  to  42 , wherein the composition is suitable for intravenous administration. 
     
     
         45 . The method of  claim 27  further comprises in step a. transfecting into the host cell: a cDNA of the second arenavirus genomic segment, a nucleic acid comprising the L protein ORF, and/or a nucleic acid comprising the NP protein ORF. 
     
     
         46 . The method of  claim 21 , wherein said administering results in a reduction in liver damage in the patient. 
     
     
         47 . The method of  claim 21 , wherein said administering results in a reduction in one or more of HBsAg, HBeAg, and HBcAg levels in the blood of the patient. 
     
     
         48 . The method of  claim 21 , wherein said administering results in a reduction in the level of an antibody against an HBV antigen in the blood of the patient. 
     
     
         49 . The viral vector of  claim 1 , wherein the pre-S2/S protein or the antigenic fragment thereof comprises an amino acid sequence encoded by the nucleotide sequence of SEQ ID NO: 1. 
     
     
         50 . The viral vector of  claim 1 , wherein the HBc protein or the antigenic fragment thereof comprises an amino acid sequence encoded by the nucleotide sequence of SEQ ID NO: 2. 
     
     
         51 . The viral vector of  claim 1 , wherein fusion of HBV HBs and HBc proteins or antigenic fragments thereof comprises an amino acid sequence encoded by the nucleotide sequence of SEQ ID NO: 3. 
     
     
         52 . The pharmaceutical composition of  claim 33 , wherein the first nucleic acid sequence is derived from LCMV and the second nucleic acid sequence is derived from Junin virus. 
     
     
         53 . The pharmaceutical composition of  claim 33 , wherein the first nucleic acid sequence is derived from Junin virus and the second nucleic acid sequence is derived from LCMV. 
     
     
         54 . The viral vector of any one of  claims 1  to  17 , wherein the arenavirus is replication-deficient and is engineered to contain a genome with the ability to amplify and express its genetic information in infected cells but unable to produce further infectious progeny particles in normal, not genetically engineered cells. 
     
     
         55 . The viral vector of any one of  claims 1  to  9 , wherein the arenavirus is replication-competent. 
     
     
         56 . The viral vector of  claim 1 , wherein the arenavirus is bisegmented. 
     
     
         57 . The viral vector of any one of  claims 1  to  17 , wherein the arenavirus is trisegmented. 
     
     
         58 . An infectious arenavirus viral vector, wherein an arenavirus open reading frame is removed and replaced by a nucleotide sequence encoding a fusion of HBV HBs and HBc proteins or antigenic fragments thereof. 
     
     
         59 . The viral vector of  claim 58 , wherein the arenavirus is lymphocytic choriomeningitis virus. 
     
     
         60 . The viral vector of  claim 58  or  59 , wherein the open reading frame that encodes the glycoprotein of the arenavirus is deleted or functionally inactivated. 
     
     
         61 . The viral vector of any one of  claims 58  to  60 , wherein the viral vector is replication-deficient. 
     
     
         62 . The viral vector of  claim 58  or  59 , wherein the viral vector is replication-competent. 
     
     
         63 . The viral vector of any one of  claim 58 ,  59 , or  62 , wherein the viral vector is trisegmented. 
     
     
         64 . A method of treating or preventing a Hepatitis B virus infection in a patient, wherein said method comprises administering to the patient the viral vector of any one of  claims 58  to  63 .