Compositions and Methods for Dengue Virus Chimeric Constructions in Vaccines

Patent metadata

Field	Value
Publication number	US-2025018006-A1
Application number	US-202418768774-A
Country	US
Kind code	A1
Filing date	Jul 10, 2024
Priority date	Mar 15, 2013
Publication date	Jan 16, 2025
Grant date	—

Abstract

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.

First claim

1 - 40 . (canceled)

41 . 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, 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.

42 . The dengue-1/dengue-2 chimera according to claim 41 , 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.

43 . The dengue-1/dengue-2 chimera according to claim 41 , wherein the polynucleotide molecule encoding the dengue-1/dengue-2 polypeptide chimera is represented by SEQ ID NO: 1 or SEQ ID NO: 13.

44 . The dengue-1/dengue-2 chimera according to claim 41 , wherein the dengue-1/dengue-2 polypeptide chimera is represented by SEQ ID NO: 2 or SEQ ID NO: 3.

45 . The dengue-1/dengue-2 chimera according to claim 41 , 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.

46 . The dengue-1/dengue-2 chimera according to claim 41 , 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).

47 . A pharmaceutical composition comprising the dengue-1/dengue-2 chimera according to claim 41 , and a pharmaceutically acceptable excipient.

48 . An immunogenic composition comprising the dengue-1/dengue-2 chimera according to claim 41 , and a pharmaceutically acceptable carrier.

49 . The immunogenic composition of claim 48 , 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.

50 . The immunogenic composition according to claim 49 , 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.

51 . The immunogenic composition according to claim 49 , wherein the polynucleotide molecule encoding the dengue-3/dengue-2 polypeptide chimera is represented by SEQ ID NO: 7 or SEQ ID NO: 15.

52 . The immunogenic composition according to claim 49 , wherein the dengue-3/dengue-2 polypeptide chimera is represented by SEQ ID NO: 8 or SEQ ID NO: 9.

53 . The immunogenic composition according to claim 52 , 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.

54 . The immunogenic composition according to claim 53 , 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.

55 . The immunogenic comp

Assignees

• Takeda Vaccines Inc
• The Government Of The United States Of America

Inventors

• Stinchcomb Dan
• Kinney Claire
• Kinney Richard M
• Livengood Jill A

Classifications

• C12N15/8613

  Chimaeric vector systems comprising heterologous sequences for production of another viral vector · CPC title

• C12N15/09

  Recombinant DNA-technology · CPC title

• C07K19/00

  Hybrid peptides {, i.e. peptides covalently bound to nucleic acids, or non-covalently bound protein-protein complexes} · CPC title

• C07K14/1825

  Flaviviruses or Group B arboviruses, e.g. yellow fever virus, japanese encephalitis, tick-borne encephalitis, dengue · CPC title

• C12N2770/24141

  Use of virus, viral particle or viral elements as a vector · CPC title