Compositions and Methods for Dengue Virus Chimeric Constructions in Vaccines

Patent metadata

Field	Value
Publication number	US-2022062375-A1
Application number	US-202117478537-A
Country	US
Kind code	A1
Filing date	Sep 17, 2021
Priority date	Mar 15, 2013
Publication date	Mar 3, 2022
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 . A modified live, attenuated dengue-2 virus strain PDK-53, wherein the modified live, attenuated dengue-2 virus strain PDK-53
is represented by a polynucleotide molecule encoding a modified live, attenuated dengue-2 virus strain PDK-53 polypeptide molecule, comprises an RNA transcribed from a cDNA comprising a polynucleotide molecule encoding the modified live, attenuated dengue-2 virus strain PDK-53 polypeptide molecule, comprises one or more polypeptide molecules encoded by a polynucleotide molecule encoding the modified live, attenuated dengue-2 virus strain PDK-53 polypeptide molecule, or 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 encoding a glutamic acid instead of a lysine in the polypeptide molecule at amino acid position 166 corresponding to prM-52, and
an adenine to guanine mutation at position 8803 encoding a valine instead of an isoleucine in the polypeptide molecule at amino acid position 2903 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 encoding a proline instead of an alanine in the polypeptide molecule at amino acid position 2129 corresponding to NS4A-36, and a cytosine to thymine mutation at position 7156 encoding a phenylalanine instead of a leucine in the polypeptide molecule at amino acid position 2354 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 . A method for inducing an immune response against dengue virus in a subject, the method comprising administering to the subject a pharmaceutically acceptable amount of the composition of claim 5 .

7 . A vector comprising 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 encoding a glutamic acid instead of a lysine in the polypeptide molecule at amino acid position 166 corresponding to prM-52, and an adenine to guanine mutation at position 8803 encoding a valine instead of an isoleucine in the polypeptide molecule at amino acid position 2903 corresponding to NS5-412.

8 . The vector according to claim 7 , wherein the vector is a plasmid vector.

9 . The vector according to claim 7 , wherein the vector is a cDNA infectious clone.

10 . An isolated cell comprising
the modified live, attenuated dengue-2 virus strain PDK-53 according to claim 1 , or a vector comprising 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 encoding a glutamic acid instead of a lysine in the polypeptide molecule at amino acid position 166 corresponding to prM-52, and an adenine to guanine mutation at position 8803 encoding a valine instead of an isoleucine in the polypeptide molecule at amino acid position 2903 corresponding to NS5-412.

11 . 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 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 encoding a glutamic acid instead of a lysine in the polypeptide molecule at amino acid position 166 corresponding to prM-52, and an adenine to guanine mutation at position 8803 encoding a valine instead of an isoleucine in the polypeptide molecule at amino acid position 2903 corresponding to NS5-412;
and
b) introducing the RNA transcribed in step a) into cells for production of the modified live, attenuated dengue-2 virus strain PDK-53.

12 . The method of claim 11 , further comprising the following step:
c) passaging the modified live, attenuated dengue-2 virus strain PDK-53 produced in step b) up to 10 times in cells.

13 . The method of claim 11 , wherein the cells are Vero cells.

14 . An immunogenic composition comprising the modified live, attenuated dengue-2 virus strain PDK-53 according to claim 1 and a pharmaceutically acceptable carrier.

15 . The immunogenic composition of claim 14 , further comprising a dengue-1/dengue-2 chimera, wherein the dengue-1/dengue-2 chimera
is represented 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 encoding a leucine instead of an isoleucine in the dengue-1/dengue-2 polypeptide chimera at amino acid position 1243 corresponding to NS2A-116;
an adenine to thymine mutation

Assignees

• Takeda Vaccines Inc
• The Government Of The Us Secretary Of The Department

Inventors

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

Classifications

• Y02A50/30

  Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change · CPC title

• C12N2770/24161

  Methods of inactivation or attenuation · CPC title

• C07K14/005 Primary

  from viruses · CPC title

• C12N2770/24134

  Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein · CPC title

• C12N2770/24122

  New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes · CPC title