Methods and compositions for chimeric coronavirus spike proteins

Patent metadata

Field	Value
Publication number	US-9884895-B2
Application number	US-201515124992-A
Country	US
Kind code	B2
Filing date	Mar 20, 2015
Priority date	Mar 20, 2014
Publication date	Feb 6, 2018
Grant date	Feb 6, 2018

Abstract

The present invention provides compositions and methods comprising a chimeric coronavirus spike protein.

First claim

What is claimed is:

1. A chimeric coronavirus spike protein comprising, in orientation from amino to carboxy terminus:
a) a first region comprising a portion of a coronavirus spike protein ectodomain that precedes a coronavirus spike protein receptor binding domain (RBD) as located in a nonchimeric coronavirus spike protein, of a first coronavirus;
b) a second region comprising a coronavirus spike protein receptor binding domain (RBD) of a second coronavirus that is different from said first coronavirus;
c) a third region comprising a portion of a coronavirus spike protein S1 domain as located in a nonchimeric coronavirus spike protein immediately downstream of the RBD, contiguous with a portion of a coronavirus spike protein S2 domain as located immediately upstream of a fusion protein domain in a nonchimeric coronavirus spike protein, wherein said third region is of said first coronavirus; and
d) a fourth region comprising a portion of a coronavirus spike protein from the start of the fusion protein domain through the carboxy terminal end as located in a nonchimeric coronavirus spike protein of a third coronavirus that is different from said first coronavirus and said second coronavirus.

2. The chimeric coronavirus spike protein of claim 1 , wherein the chimeric coronavirus spike protein is derived from subgroup 1a coronaviruses, subgroup 1b coronaviruses, subgroup 2a coronaviruses, subgroup 2b coronaviruses, subgroup 2c coronaviruses, subgroup 2d coronaviruses or subgroup 3 coronaviruses.

3. The chimeric coronavirus of claim 2 , derived from subgroup 2b coronaviruses wherein said first, second and third subgroup 2b coronaviruses are different from one another and wherein the subgroup 2b coronaviruses are selected from the group consisting of Bat SARS CoV (GenBank Accession No. FJ211859), SARS CoV (GenBank Accession No. FJ211860), BtSARS.HKU3.1 (GenBank Accession No. DQ022305), BtSARS.HKU3.2 (GenBank Accession No. DQ084199), BtSARS.HKU3.3 (GenBank Accession No. DQ084200), BtSARS.Rm1 (GenBank Accession No. DQ412043), BtCoV.279.2005 (GenBank Accession No. DQ648857), BtSARS.Rf1 (GenBank Accession No. DQ412042), BtCoV.273.2005 (GenBank Accession No. DQ648856), BtSARS.Rp3 (GenBank Accession No. DQ071615), SARS CoV.A022 (GenBank Accession No. AY686863), SARSCoV.CUHK-W1 (GenBank Accession No. AY278554), SARSCoV.GD01 (GenBank Accession No. AY278489), SARSCoV.HC.SZ.61.03 (GenBank Accession No. AY515512), SARSCoV.SZ16 (GenBank Accession No. AY304488), SARSCoV.Urbani (GenBank Accession No. AY278741), SARSCoV.civet010 (GenBank Accession No. AY572035), and SARSCoV.MA.15 (GenBank Accession No. DQ497008).

4. The chimeric subgroup 2b coronavirus spike protein of claim 3 , wherein said first subgroup 2b coronavirus is Bat SARS CoV-HKU3 (GenBank Accession No. FJ211859), said second subgroup 2b coronavirus is SARSCoV.Urbani (GenBank Accession No. AY278741.1), and said third subgroup 2b coronavirus is BtCoV 279.2005 (DQ648857).

5. The chimeric coronavirus spike protein of claim 1 , comprising the amino acid sequence:

(SEQ ID NO: 1)

1

MKILIFAFLA NLAKAQEGCG IISRKPQPKM AQVSSSRRGV YYNDDIFRSD VLHLTQDYFL

61

PFDSNLTQYF SLNVDSDRYT YFDNPILDFG DGVYFAATEK SNVIRGWIFG SSFDNTTQSA

121

VIVNNSTHII IRVCNFNLCK EPMYTVSRGT QQNAWVYQSA FNCTYDRVEK SFQLDTTPKT

181

GNFKDLREYV FKNRDGFLSV YQTYTAVNLP RGLPTGFSVL KPILKLPFGI NITSYRVVMA

241

MFSQTTSNFL PESAAYYVGN LKYSTFMLRF NENGTITDAV DCSQNPLAEL KCTIKNFNVD

301

KGIYQTSNFR VSPTQEVIRF PNITN
LCPFG EVFNATKFPS VYAWERKKIS NCVADYSVLY

361

NSTFFSTFKC YGVSATKLND LCFSNVYADS FVVKGDDVRQ IAPGQTGVIA DYNYKLPDDF

421

MGCVLAWNTR NIDATSTGNY NYKYRYLRHG KLRPFERDIS NVPFSPDGKP CTPPALNCYW

481

PLNDYGFYTT TGIGYQPYRV VVLS
FELLNA PATVCGPKLS TDLVKNQCVN FNFNGLKGTG

541

VLTSSSKRFQ SFQQFGRDTS DFTDSVRDPQ TLEILDISPC SFGGVSVITP GTNASSEVAV

601

LYQDVNCTDV PTAIRADQLT PAWRVYSTGV NVFQTQAGCL IGAEHVNASY ECDIPIGAGI

661

CASYHTASVL RSTGQKSIVA YTMSLGAENS IAYANNSIAI PTNFSISVTT EVMPVSMAKT

721

AVDCTMYICG DSLECSNLLL QYGSFCTQLN RALTGIAIEQ DKNTQEVFAQ VKQMYKTPAI

781

KDFGGFNFSQ ILPDPSKPTK RSFIEDLLFN KVTLADAGFM KQYGDCLGDV SARDLICAQK

841

FNGLTVLPPL LTDEMVAAY
T AALVSGTATA GWTFGAGSAL QIPFAMQMAY RFNGIGVTQN

901

VLYENQKQIA NQFNKAISQI QESLTTTSTA LGKLQDVVND NAQALNTLVK QLSSNFGAIS

961

SVLNDILSRL DKVEAEVQID RLITGRLQSL QTYVTQQLIR AAEIRASANL AATKMSECVL

1021

GQSKRVDFCG KGYHLMSFPQ AAPHGVVFLH VTYVPSQERN FTTAPAICHE GKAYFPREGV

1081

FVSNGTSWFI TQRNFYSPQI ITTDNTFVAG NCDVVIGIIN NTVYDPLQPE LDSFKEELDK

1141

YFKNHTSPDV DLGDISGINA SVVNIQKEID RLNEVAKNLN ESLIDLQELG KYEQYIKWPW

1201
YVWLGFIAGL IAIVMVTILL CCMTSCCSCL KGACSCGSCC KFDEDDSEPV LKGVKLHYT .

Assignees

• Univ North Carolina Chapel Hill

Inventors

• Baric Ralph
• Agnihothram Sudhakar
• Yount Boyd

Classifications

• C12N2770/36144

  Chimeric viral vector comprising heterologous viral elements for production of another viral vector · CPC title

• A61K2039/5256

  expressing foreign proteins · CPC title

• A61P31/12

  Antivirals · CPC title

• C07K2317/76

  Antagonist effect on antigen, e.g. neutralization or inhibition of binding · CPC title

• C12N2770/20022

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