Serial No. <Unassigned>
`Int’?] Application No: PCT/EP2020/075541
`Docket No. KATO060PA/P186468USPC
`Date: March 14, 2022
`Preliminary Amendment at National-Stage Entry
`
`AMENDMENTS TO THE CLAIMS
`
`This listing of claims replaces all prior versions andlistings of claims in the application.
`
`Listing of Claims
`
`1-15.
`
`(Canceled)
`
`16.
`
`(New) A polynucleotide comprising a sequence ofa live, infectious, attenuated
`
`Flavivirus, the polynucleotide comprising:
`
`a nucleotide sequence encodingat least a part of an arenavirus glycoprotein, the
`
`arenavirus glycoprotein comprising a GP1 domain and a GP2 domain,the
`
`arenavirus glycoprotein lacking an N terminal signal peptide and a GP2
`
`transmembrane domain,
`
`wherein:
`
`the nucleotide sequenceis located at an intergenic region between an E geneofthe
`
`Flavivirus and an NS1 gene ofthe Flavivirus, the E gene encoding an E protein,
`
`the NS1 gene encoding an NS1 protein, the NS1 protein comprising a signal
`
`peptide, such that a chimeric virus is expressed; and
`
`the nucleotide sequenceis translatable such that the nucleotide sequence encodes a
`
`chimeric viral peptide, the chimeric viral peptide comprising:
`
`(a)
`
`a further signal peptide, positioned C terminally of the E protein, wherein
`
`the further signal peptide has the same sequenceasthe signal peptide of
`
`the NS1 protein;
`
`(b)
`
`the arenavirus glycoprotein, positioned C terminally of the further signal
`
`peptide; and
`
`(c)
`
`a TM1 and a TM2 domainofa further flaviviral E protein, positioned C
`
`terminally of the arenavirus glycoprotein; and
`
`the chimeric viral peptide is positioned N terminally of the NS1 protein of the Flavivirus.
`
`Page 4 of 12
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`Serial No. <Unassigned>
`Int’?] Application No: PCT/EP2020/075541
`Docket No. KATO060PA/P186468USPC
`Date: March 14, 2022
`Preliminary Amendment at National-Stage Entry
`
`17.
`
`(New) The polynucleotide according to claim 16, wherein the Flavivirus is Yellow Fever
`
`virus.
`
`18.
`
`(New) The polynucleotide according to claim 17, wherein the Yellow Fevervirusis the
`
`YF17Dstrain.
`
`19.
`
`(New) The polynucleotide according to claim 16, wherein the arenavirus glycoprotein is
`
`a Mammarenavirus glycoprotein.
`
`20.
`
`(New) The polynucleotide according to claim 16, wherein the arenavirus glycoprotein is
`
`a Lassa virus glycoprotein.
`
`21.
`
`(New) The polynucleotide according to claim 16, wherein the sequence of the chimeric
`
`viral peptide comprises the amino acid sequenceset forth in SEQ ID NO:2 or SEQ ID NO:4.
`
`22.
`
`(New) The polynucleotide according to claim 16, wherein the arenavirus glycoprotein
`
`comprises a R207C stabilizing mutation, a G360C stabilizing mutation, and an E329Pstabilizing
`
`mutation.
`
`23.
`
`(New) The polynucleotide according to claim 16, wherein the arenavirus glycoprotein
`
`comprises a R246A proteolytic cleavagesite.
`
`24.
`
`(New) The polynucleotide according to claim 16, wherein the further signal peptide
`
`comprises the sequenceset forth by SEQ ID NO:10.
`
`25.
`
`(New) The polynucleotide according to claim 16, wherein the chimeric viral peptide
`
`further comprises:
`
`the TM1 domain having the sequenceset forth in SEQ ID NO:14; or
`
`the TM2 domain having the sequenceset forth in SEQ ID NO:15.
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`Page 5 of 12
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`Serial No. <Unassigned>
`Int’?] Application No: PCT/EP2020/075541
`Docket No. KATO060PA/P186468USPC
`Date: March 14, 2022
`Preliminary Amendment at National-Stage Entry
`
`26.
`
`(New) The polynucleotide according to claim 16, further comprising a junction
`
`sequence, wherein the junction sequenceis selected from the group consisting of:
`
`the junction sequenceset forth in SEQ ID NO:11, positioned between the further signal
`
`peptide sequence and the GP1 domain;
`
`the junction sequenceset forth in SEQ ID NO:12, positioned between the GP2 domain
`
`and the TM1 domain; and
`
`the junction sequenceset forth in SEQ ID NO:13, positioned between the TM2 domain
`
`and the NS1 protein.
`
`27.
`
`(New) The polynucleotide according to claim 16, wherein the nucleotide sequence
`
`comprises:
`
`a nucleotide sequence that is 95% identical to the nucleotide sequence set forth in
`
`SEQ ID NO:1, or
`
`a nucleotide sequence that is 95% identical to the nucleotide sequence set forth in SEQ
`
`ID NO:3.
`
`28.
`
`(New) The polynucleotide according to claim 27, wherein the nucleotide sequence
`
`further comprises portions of the nucleotide sequence that encode the peptide sequences set forth
`
`in SEQ ID NO:11, SEQ ID NO:12, and SEQ ID NO:13.
`
`29.
`
`(New) A pharmaceutical composition comprising a polynucleotide according to claim 16
`
`and at least one pharmaceutically acceptable carrier, diluent, or excipient.
`
`30.
`
`(New) A method of vaccinating an individual against an arenavirus infection, the method
`
`comprising administering to the individual the polynucleotide according to claim 16.
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`Page 6 of 12
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`Serial No. <Unassigned>
`Int’?] Application No: PCT/EP2020/075541
`Docket No. KATO060PA/P186468USPC
`Date: March 14, 2022
`Preliminary Amendment at National-Stage Entry
`
`31.
`
`(New) A chimeric, live, infectious, attenuated Flavivirus comprising:
`
`an E protein;
`
`an NS1 protein comprising a signal peptide; and
`
`a chimeric protein comprising:
`
`at least a part of an arenavirus Glycoprotein, the arenavirus glycoprotein
`
`comprising a GP1 domain and a GP2 domain, wherein the arenavirus
`
`glycoprotein lacks an N terminal signal peptide and a GP2 transmembrane
`
`domain, and the arenavirus glycoprotein is located between the E protein
`
`and the NS1 protein;
`
`a further signal peptide, positioned C terminally of the E protein, wherein the
`
`further signal peptide has the same sequenceasthe signal peptide of the
`
`NS1 protein, and wherein the arenavirus glycoprotein is positioned C
`
`terminally of the further signal peptide; and
`
`a TM1 domain and a TM2 domainofa further Flaviviral E protein, positioned C
`
`terminally of the arenavirus glycoprotein,
`
`wherein the chimeric protein is positioned N terminally of the NS1 protein.
`
`32.
`
`(New) The chimeric, live, infectious, attenuated Flavivirus accordingto claim 31,
`
`wherein the Flavivirus is Yellow Fever Virus.
`
`33.
`
`(New) The chimeric, live, infectious, attenuated Flavivirus accordingto claim 31,
`
`wherein the arenavirus glycoprotein is Lassa virus glycoprotein.
`
`34.
`
`(New) A pharmaceutical composition comprising a chimeric, live, infectious, attenuated
`
`Flavivirus according to claim 31 and at least one pharmaceutically acceptable carrier, diluent, or
`
`excipient.
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`Page 7 of 12
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`Serial No. <Unassigned>
`Int’?] Application No: PCT/EP2020/075541
`Docket No. KATO060PA/P186468USPC
`Date: March 14, 2022
`Preliminary Amendment at National-Stage Entry
`
`35.
`
`(New) A method of vaccinating an individual against an arenavirus infection, the method
`
`comprising administering to the individual the chimeric, live, infectious, attenuated Flavivirus
`
`according to claim 31.
`
`Page 8 of 12
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