Compositions and methods for dengue virus chimeric constructions in vaccines
Embodiments herein report compositions, uses and manufacturing of dengue virus constructs and live attenuated dengue viruses. Some embodiments concern a composition that includes, but is not limited to, a tetravalent dengue virus composition. In certain embodiments, compositions can include constructs of one or more serotypes of dengue virus, such as dengue-1 (DEN-1) virus, dengue-2 (DEN-2) virus, dengue-3 (DEN-3) or dengue-4 (DEN-4) virus constructs. In other embodiments, constructs disclosed herein can be combined in a composition to generate a vaccine against more one or more dengue virus constructs that may or may not be subsequently passaged in mammalian cells.
1. A modified live, attenuated dengue-2 virus strain PDK-53, wherein the modified live, attenuated dengue-2 virus strain PDK-53
is encoded by a polynucleotide molecule encoding a modified live, attenuated dengue-2 virus strain PDK-53 polypeptide molecule,
wherein the polynucleotide molecule encoding the modified live, attenuated dengue-2 virus strain PDK-53 polypeptide molecule comprises at least one mutation, wherein the at least one mutation comprises:
an adenine to guanine mutation at position 592 in the numbering of SEQ ID NO: 14 encoding a glutamic acid instead of a lysine in the polypeptide molecule at amino acid position 166 in the numbering of SEQ ID NO: 6 corresponding to prM-52, and
an adenine to guanine mutation at position 8803 in the numbering of SEQ ID NO: 14 encoding a valine instead of an isoleucine in the polypeptide molecule at amino acid position 2903 in the numbering of SEQ ID NO: 6 corresponding to NS5-412.
2. The modified live, attenuated dengue-2 virus strain PDK-53 according to claim 1 , wherein the polynucleotide molecule encoding the modified live, attenuated dengue-2 virus strain PDK-53 polypeptide molecule further comprises at least one additional mutation of:
a guanine to cytosine mutation at nucleic acid position 6481 in the numbering of SEQ ID NO: 14 encoding a proline instead of an alanine in the polypeptide molecule at amino acid position 2129 in the numbering of SEQ ID NO: 6 corresponding to NS4A-36, and
a cytosine to thymine mutation at position 7156 in the numbering of SEQ ID NO: 14 encoding a phenylalanine instead of a leucine in the polypeptide molecule at amino acid position 2354 in the numbering of SEQ ID NO: 6 corresponding to NS4B-111.
3. The modified live, attenuated dengue-2 virus strain PDK-53 according to claim 1 , wherein the polynucleotide molecule encoding the modified live, attenuated dengue-2 virus strain PDK-53 polypeptide molecule is represented by SEQ ID NO: 4 or SEQ ID NO: 14.
4. The modified live, attenuated dengue-2 virus strain PDK-53 according to claim 1 , wherein the modified, live attenuated dengue-2 virus strain PDK-53 polypeptide molecule is represented by SEQ ID NO: 5 or SEQ ID NO: 6.
5. A pharmaceutical composition comprising the modified live, attenuated dengue-2 virus strain PDK-53 according to claim 1 , and a pharmaceutically acceptable excipient.
6. An immunogenic composition comprising the modified live, attenuated dengue-2 virus strain PDK-53 according to claim 1 , and a pharmaceutically acceptable carrier.
7. The immunogenic composition of claim 6 , further comprising a dengue-1/dengue-2 chimera, wherein the dengue-1/dengue-2 chimera
is encoded by a polynucleotide molecule encoding a dengue-1/dengue-2 polypeptide chimera,
comprises an RNA transcribed from a cDNA comprising a polynucleotide molecule encoding the dengue-1/dengue-2 polypeptide chimera, comprises one or more polypeptide molecules encoded by a polynucleotide molecule encoding the dengue-1/dengue-2 polypeptide chimera, or
is obtainable by a method for producing a dengue-1/dengue-2 chimera, the method comprising the following steps:
a) transcribing an RNA from a cDNA comprising a polynucleotide molecule encoding the dengue-1/dengue-2 polypeptide chimera, and
b) introducing the RNA transcribed in step a) into cells for production of the dengue-1/dengue-2 chimera,
wherein the polynucleotide molecule encoding the dengue-1/dengue-2 polypeptide chimera comprises a first nucleotide sequence encoding nonstructural proteins from a modified live, attenuated dengue-2 virus strain PDK 53, a second nucleotide sequence encoding at least one structural protein from dengue-1, and at least one mutation, wherein the at least one mutation comprises one or more of:
an adenine to cytosine mutation at position 3823 in the numbering of SEQ ID NO: 13 encoding a leucine instead of an isoleucine in the dengue-1/dengue-2 polypeptide chimera at amino acid position 1243 in the numbering of SEQ ID NO: 3 corresponding to NS2A-116;
an adenine to thymine mutation at position 4407 in the numbering of SEQ ID NO: 13 encoding an aspartic acid instead of a glutamic acid in the dengue-1/dengue-2 polypeptide chimera at amino acid position 1437 in the numbering of SEQ ID NO: 3 corresponding to NS2B-92; and
an adenine to guanine mutation at position 7311.
8. The immunogenic composition according to claim 7 , wherein the polynucleotide molecule encoding the dengue-1/dengue-2 polypeptide chimera further comprises at least one additional mutation of:
a cytosine to thymine mutation at position 7148 in the numbering of SEQ ID NO: 13 encoding an isoleucine instead of a threonine in the dengue-1/dengue-2 polypeptide chimera at amino acid position 2351 in the numbering of SEQ ID NO: 3 corresponding to NS4B-108; and
a guanine to cytosine mutation at position 2384 in the numbering of SEQ ID NO: 13 encoding an alanine instead of glycine in the dengue-1/dengue-2 polypeptide chimera at amino acid position 763 in the numbering of SEQ ID NO: 3 corresponding to E-483.
9. The immunogenic composition according to claim 7 , wherein the polynucleotide molecule encoding the dengue-1/dengue-2 polypeptide chimera is represented by SEQ ID NO: 1 or SEQ ID NO: 13.
10. The immunogenic composition according to claim 7 , wherein the dengue-1/dengue-2 polypeptide chimera is represented by SEQ ID NO: 2 or SEQ ID NO: 3.
11. The immunogenic composition according to claim 7 , wherein the nonstructural proteins from the modified live, attenuated dengue-2 virus strain PDK 53 are selected from the group consisting of NS1, NS2A, NS2B, NS3, NS4A, NS4B and NS5.
12. The immunogenic composition according to claim 7 , wherein the at least one structural protein from dengue- 1 is selected from the group consisting of capsid protein (C), premembrane/membrane protein (prM) and envelope protein (E).
13. The immunogenic composition according to claim 7 , further comprising a dengue-3/dengue-2 chimera, wherein the dengue-3/dengue-2 chimera
is encoded by a polynucleotide molecule encoding a dengue-3/dengue-2 polypeptide chimera,
comprises an RNA transcribed from a cDNA comprising a polynucleotide molecule encoding the dengue-3/dengue-2 polypeptide chimera,
comprises one or more polypeptide molecules encoded by a polynucleotide molecule encoding the dengue-3/dengue-2 polypeptide chimera, or
is obtainable by a method for producing a dengue-3/dengue-2 chimera, the method comprising the following steps:
a) transcribing an RNA from a cDNA comprising a polynucleotide molecule encoding the dengue-3/dengue-2 polypeptide chimera, and
b) introducing the RNA transcribed in step a) into cells for production of the dengue-3/dengue-2 chimera,
wherein the polynucleotide molecule encoding the dengue-3/dengue-2 polypeptide chimera comprises a first nucleotide sequence encoding nonstructural proteins from a modified live, attenuated dengue-2 virus strain PDK-53, a second nucleotide sequence encoding at least one structural protein from dengue-3, and at least one mutation, wherein the at least one mutation comprises one or more of:
an adenine to thymine mutation at position 1603 in the numbering of SEQ ID NO: 15 encoding a serine instead of a threonine in the dengue-3/dengue-2 polypeptide chimera at amino acid position 503 in the numbering of SEQ ID NO: 9 corresponding to E-223; and
an adenine to guanine mutation at position 7620 in the number of SEQ ID NO: 15.
14. The immunogenic composition according to claim 13 , wherein the polynucleotide molecule encoding the dengue-3/dengue-2 polypeptide chimera further comprises a guanine to adenine mutation at position 6436 in the numbering of SEQ ID NO: 15 encoding an asparagine instead of an aspartic acid in the dengue-3/dengue-2 polypeptide chimera at amino acid position 2114 in the numbering of SEQ ID NO: 9 corresponding to NS4A-23.
15. The immunogenic composition according to claim 13 , wherein the polynucleotide molecule encoding the dengue-3/dengue-2 polypeptide chimera is represented by SEQ ID NO: 7 or SEQ ID NO: 15.
16. The immunogenic composition according to claim 13 , wherein the dengue-3/dengue-2 polypeptide chimera is represented by SEQ ID NO: 8 or SEQ ID NO: 9.
17. The immunogenic composition according to claim 16 , further comprising a dengue-4/dengue-2 chimera, wherein the dengue-4/dengue-2 chimera
is encoded by a polynucleotide molecule encoding a dengue-4/dengue-2 polypeptide chimera,
comprises an RNA transcribed from a cDNA comprising a polynucleotide molecule encoding the dengue-4/dengue-2 polypeptide chimera,
comprises one or more polypeptide molecules encoded by a polynucleotide molecule encoding the dengue-4/dengue-2 polypeptide chimera, or
is obtainable by a method for producing a dengue-4/dengue-2 chimera, the method comprising the following steps:
a) transcribing an RNA from a cDNA comprising a polynucleotide molecule encoding the dengue-4/dengue-2 polypeptide chimera, and
b) introducing the RNA transcribed in step a) into cells for production of the dengue-4/dengue-2 chimera,
wherein the polynucleotide molecule encoding the dengue-4/dengue-2 polypeptide chimera comprises a first nucleotide sequence encoding nonstructural proteins from a modified live, attenuated dengue-2 virus strain PDK-53, a second nucleotide sequence encoding at least one structural protein from dengue-4, and at least one mutation, wherein the at least one mutation comprises one or more of:
an adenine to thymine mutation at position 225 in the numbering of SEQ ID NO: 16;
an adenine to guanine mutation at position 3674 in the numbering of SEQ ID NO: 16 encoding a glycine instead of an aspartic acid in the dengue-4/dengue-2 polypeptide chimera at amino acid position 1193 in the numbering of SEQ ID NO: 12 corresponding to NS2A-66;
a cytosine to thymine mutation at position 5391 in the numbering of SEQ ID NO: 16;
a cytosine to thymine mutation at position 6437 in the numbering of SEQ ID NO: 16 encoding a valine instead of an alanine in the dengue-4/dengue-2 polypeptide chimera at amino acid position 2114 in the numbering of SEQ ID NO: 12 corresponding to NS4A-21, and an adenine to cytosine mutation at position 9750 in the numbering of SEQ ID NO: 16.
18. The immunogenic composition according to claim 17 , wherein the polynucleotide molecule encoding the dengue-4/dengue-2 polypeptide chimera further comprises a thymine to cytosine mutation at position 7026 in the numbering of SEQ ID NO: 16.
19. The immunogenic composition according to claim 17 , wherein the polynucleotide molecule encoding the dengue-4/dengue-2 polypeptide chimera further comprises an adenine to guanine mutation at position 3773 in the numbering of SEQ ID NO: 16 encoding an arginine instead of a lysine in the dengue-4/dengue-2 polypeptide chimera at amino acid position 1226 in the numbering of SEQ ID NO: 12 corresponding to NS2A-99.
20. The immunogenic composition according to claim 17 , wherein the polynucleotide molecule encoding the dengue-4/dengue-2 polypeptide chimera further comprises a cytosine to thymine mutation at position 7538 in the numbering of SEQ ID NO: 16 encoding a phenylalanine instead of a serine in the dengue-4/dengue-2 polypeptide chimera at amino acid position 2481 in the numbering of SEQ ID NO: 12 corresponding to NS4B-238.
21. The immunogenic composition according to claim 17 , wherein the polynucleotide molecule encoding the dengue-4/dengue-2 polypeptide chimera is represented by SEQ ID NO: 10 or SEQ ID NO: 16.
22. The immunogenic composition according to claim 17 , wherein the dengue 4/dengue-2 polypeptide chimera is represented by SEQ ID NO: 11 or SEQ ID NO: 12.
23. A modified live, attenuated dengue-2 virus strain PDK-53, wherein the modified live, attenuated dengue-2 virus strain PDK-53 comprises an RNA transcribed from a cDNA comprising a polynucleotide molecule encoding the modified live, attenuated dengue-2 virus strain PDK-53 polypeptide molecule, wherein the polynucleotide molecule encoding the modified live, attenuated dengue-2 virus strain PDK-53 polypeptide molecule comprises at least one mutation, wherein the at least one mutation comprises:
an adenine to guanine mutation at position 592 in the numbering of SEQ ID NO: 14 encoding a glutamic acid instead of a lysine in the polypeptide molecule at amino acid position 166 in the numbering of SEQ ID NO: 6 corresponding to prM-52, and
an adenine to guanine mutation at position 8803 in the numbering of SEQ ID NO: 14 encoding a valine instead of an isoleucine in the polypeptide molecule at amino acid position 2903 in the numbering of SEQ ID NO: 6 corresponding to NS5-412.
24. A modified live, attenuated dengue-2 virus strain PDK-53, wherein the modified live, attenuated dengue-2 virus strain PDK-53 comprises one or more polypeptide molecules encoded by a polynucleotide molecule encoding the modified live, attenuated dengue-2 virus strain PDK-53 polypeptide molecule,
wherein the polynucleotide molecule encoding the modified live, attenuated dengue-2 virus strain PDK-53 polypeptide molecule comprises at least one mutation, wherein the at least one mutation comprises:
an adenine to guanine mutation at position 592 in the numbering of SEQ ID NO: 14 encoding a glutamic acid instead of a lysine in the polypeptide molecule at amino acid position 166 in the numbering of SEQ ID NO: 6 corresponding to prM-52, and
an adenine to guanine mutation at position 8803 in the numbering of SEQ ID NO: 14 encoding a valine instead of an isoleucine in the polypeptide molecule at amino acid position 2903 in the numbering of SEQ ID NO: 6 corresponding to NS5-412.
25. A modified live, attenuated dengue-2 virus strain PDK-53, wherein the modified live, attenuated dengue-2 virus strain PDK-53 is obtainable by a method for producing a modified live, attenuated dengue-2 virus strain PDK-53, the method comprising the following steps:
a) transcribing an RNA from a cDNA comprising a polynucleotide molecule encoding the modified live, attenuated dengue-2 virus strain PDK-53 polypeptide molecule, and
b) introducing the RNA transcribed in step a) into cells for production of the modified live, attenuated dengue-2 virus strain PDK-53,
wherein the polynucleotide molecule encoding the modified live, attenuated dengue-2 virus strain PDK-53 polypeptide molecule comprises at least one mutation, wherein the at least one mutation comprises:
an adenine to guanine mutation at position 592 in the numbering of SEQ ID NO: 14 encoding a glutamic acid instead of a lysine in the polypeptide molecule at amino acid position 166 in the numbering of SEQ ID NO: 6 corresponding to prM-52, and
an adenine to guanine mutation at position 8803 in the numbering of SEQ ID NO: 14 encoding a valine instead of an isoleucine in the polypeptide molecule at amino acid position 2903 in the numbering of SEQ ID NO: 6 corresponding to NS5-412.
26. The immunogenic composition according to claim 13 , wherein the nonstructural proteins from the modified live, attenuated dengue-2 virus strain PDK 53 are selected from the group consisting of NS1, NS2A, NS2B, NS3, NS4A, NS4B and NS5.
27. The immunogenic composition according to claim 13 , wherein the at least one structural protein from dengue-3 is selected from the group consisting of capsid protein (C), premembrane/membrane protein (prM) and envelope protein (E).
28. The immunogenic composition according to claim 17 , wherein the nonstructural proteins from the modified live, attenuated dengue-2 virus strain PDK 53 are selected from the group consisting of NS1, NS2A, NS2B, NS3, NS4A, NS4B and NS5.
29. The immunogenic composition according to claim 17 , wherein the at least one structural protein from dengue-4 is selected from the group consisting of capsid protein (C), premembrane/membrane protein (prM) and envelope protein (E).