Stabilized Filovirus glycoprotein trimers
AssigneeJanssen Vaccines & Prevention B.V.
InventorsJohannes Langedijk, Lucy Rutten, Sven Blokland
Life of this patent
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Abstract
Filovirus glycoprotein mutations that stabilize the trimeric form of the glycoprotein are provided. The Filovirus glycoproteins have certain amino acid substitutions at specified positions in the glycoprotein sequence. The Filovirus glycoproteins described herein have an improved percentage of trimer formation and/or an improved trimer yield as compared to a Filovirus glycoprotein that does not have one or more of the indicated amino acid substitutions. Also provided are nucleic acid molecules and vectors encoding the Filovirus glycoproteins, as well as compositions containing the Filovirus glycoproteins, nucleic acid, and vectors.
Claims21
A recombinant glycoprotein obtained by replacing an amino acid residue of a Filovirus glycoprotein with a non-charged amino acid residue,
- wherein the Filovirus glycoprotein is selected from the group consisting of:
- 1) A Zaire Ebolavirus Mayinga glycoprotein comprising the amino acid sequence of SEQ ID NO:
- 2 or a Zaire Ebolavirus Mayinga glycoprotein with a deletion between amino acid residues 288 and 444, comprising the amino acid sequence of SEQ ID NO:
- 6;
- wherein the recombinant glycoprotein is obtained by replacing a lysine at position 556 with a hydrophobic amino acid residue selected from the group consisting of F, I, L, M, V, W, and Y;
- wherein the numbering of the positions is according to the numbering of the glycoprotein of SEQ ID NO:
- 2;
- 2) A Zaire Ebolavirus Makona GP glycoprotein comprising the amino acid sequence of SEQ ID NO:
- 10 or a Zaire Ebolavirus Makona GP glycoprotein with a deletion between amino acid residues 282 and 440, comprising the amino acid sequence of SEQ ID NO:
- 14;
- wherein the recombinant glycoprotein is obtained by replacing a lysine at position 556 with a hydrophobic amino acid residue selected from the group consisting of F, I, L, M, V, W, and Y;
- wherein the numbering of the positions is according to the numbering of the glycoprotein of SEQ ID NO:
- 10;
- 3) A Zaire Ebolavirus Kikwit glycoprotein comprising the amino acid sequence of SEQ ID NO:
- 19 or a Zaire Ebolavirus Kikwit glycoprotein with a deletion between amino acid residues 282 and 440, comprising the amino acid sequence of SEQ ID NO:
- 23;
- wherein a lysine at position 556 has been replaced with a hydrophobic amino acid residue selected from the group consisting of F, I, L, M, V, W, and Y;
- wherein the numbering of the positions is according to the numbering of the glycoprotein of SEQ ID NO:
- 19;
- and
- 4) a Marburg glycoprotein with a deleted mucin-like domain, comprising the amino acid sequence of SEQ ID NO:
- 27,
- wherein a histidine at position 402 has been replaced with a hydrophobic amino acid residue selected from the group consisting of F, I, L, M, V, W, and Y;
- wherein the numbering of the positions is according to the numbering of the glycoprotein of SEQ ID NO:
- 27.
A recombinant Filovirus glycoprotein, comprising a Zaire Ebolavirus Mayinga glycoprotein comprising the amino acid sequence of SEQ ID NO:
- 2 or a Zaire Ebolavirus Mayinga glycoprotein with a deletion between amino acid residues 288 and 444, comprising the amino acid sequence of SEQ ID NO:
- 6;
- wherein the recombinant glycoprotein is obtained by replacing a lysine at position 556 with a hydrophobic amino acid residue selected from the group consisting of F, I, L, M, V, W, and Y;
- wherein the numbering of the positions is according to the numbering of the glycoprotein of SEQ ID NO:
- 2.
A method of improving the trimer formation of a Filovirus glycoprotein, the method comprising substituting a lysine amino acid residue at position 556 of the Filovirus glycoprotein for a hydrophobic amino acid residue selected from the group consisting of F, I, L, M, V, W, and Y, wherein:
- the Filovirus glycoprotein is a Zaire Ebolavirus Mayinga glycoprotein comprising the amino acid sequence of SEQ ID NO:
- 2 or a Zaire Ebolavirus Mayinga glycoprotein with a deletion between amino acid residues 288 and 444, comprising the amino acid sequence of SEQ ID NO:
- 6;
- and
- the numbering of the positions is according to the numbering of the glycoprotein of SEQ ID NO:
- 2;
- wherein the substitution at position 556 increases the percentage of trimer formation.
A composition comprising:
- a)
- a recombinant Filovirus glycoprotein, comprising a Zaire Ebolavirus Mayinga glycoprotein comprising the amino acid sequence of SEQ ID NO:
- 2 or a Zaire Ebolavirus Mayinga glycoprotein with a deletion between amino acid residues 288 and 444, comprising the amino acid sequence of SEQ ID NO:
- 6;
- wherein the recombinant glycoprotein is obtained by replacing a lysine at position 556 with a hydrophobic amino acid residue selected from the group consisting of F, I, L, M, V, W, and Y;
- wherein the numbering of the positions is according to the numbering of the glycoprotein of SEQ ID NO:
- 2;
- and
- b) a pharmaceutically acceptable carrier.
A composition comprising:
- a) a trimeric complex comprising a noncovalent oligomer of three recombinant Filovirus glycoproteins, each recombinant Filovirus glycoprotein comprising a Zaire Ebolavirus Mayinga glycoprotein comprising the amino acid sequence of SEQ ID NO:
- 2 or a Zaire Ebolavirus Mayinga glycoprotein with a deletion between amino acid residues 288 and 444, comprising the amino acid sequence of SEQ ID NO:
- 6;
- wherein the recombinant glycoprotein is obtained by replacing a lysine at position 556 with a hydrophobic amino acid residue selected from the group consisting of F, I, L, M, V, W, and Y;
- wherein the numbering of the positions is according to the numbering of the glycoprotein of SEQ ID NO:
- 2;
- and
- b) a pharmaceutically acceptable carrier.
A composition comprising:
- a) a particle displaying on its surface a recombinant Filovirus glycoprotein comprising a Zaire Ebolavirus Mayinga glycoprotein comprising the amino acid sequence of SEQ ID NO:
- 2 or a Zaire Ebolavirus Mayinga glycoprotein with a deletion between amino acid residues 288 and 444, comprising the amino acid sequence of SEQ ID NO:
- 6;
- wherein the recombinant glycoprotein is obtained by replacing a lysine at position 556 with a hydrophobic amino acid residue selected from the group consisting of F, I, L, M, V, W, and Y;
- wherein the numbering of the positions is according to the numbering of the glycoprotein of SEQ ID NO:
- 2
- wherein the particle is a liposome, a virus-like particle (VLP), a nanoparticle, a virosome, or an exosome;
- and
- a) a pharmaceutically acceptable carrier.
Dependencies are inferred from the claim text.