IP Library Granted Patent US 12,365,879
Granted Patent B2
US 12,365,879 · App. 17/827,320 · Granted Jul 22, 2025

Synthetic chimeric poxviruses

Inventors: David Evans (Edmonton, CA); Ryan Noyce (Edmonton, CA); Seth Lederman (Austin, TX)
Assignee: TONIX PHARMA LIMITED
C12N7/00A61K39/12A61K39/275A61P31/20A61K2039/5254C12N2710/24021C12N2710/24034C12N2710/24121C12N2710/24134
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Quick Facts
Patent No.
US 12,365,879
App. No.
17/827,320
Granted
Jul 22, 2025
Kind
B2
Abstract

The invention relates, in general, to synthetic chimeric poxviruses, compositions comprising such viruses, and the development and use of systems and methods for producing such synthetic chimeric poxviruses. The synthetic chimeric poxviruses are well suited for live virus vaccines and pharmaceutical formulations.

Claims (33)

1. A method of producing a synthetic chimeric orthopoxvirus (scOPV) comprising the steps of:

(i) chemically synthesizing overlapping DNA fragments that correspond to substantially all of a wild type genome selected from the group consisting of a camelpox virus, a cowpox virus, an ectromelia virus, a horsepox virus, a monkeypox virus, a vaccinia virus, a variola virus, a rabbitpox virus, a raccoon poxvirus, a skunkpox virus, a Taterapox virus, an Uasin Gishu disease virus, and a volepox virus;

(ii) chemically synthesizing terminal hairpin loops independently derived from an orthopoxvirus and ligating the terminal hairpin loops derived from the chemically synthesized hairpin loops to the left and right termini of the overlapping DNA fragments derived from the chemically synthesized DNA of step (i), and wherein the orthopoxvirus is selected from the group consisting of a camelpox virus, a cowpox virus, an ectromelia virus, a horsepox virus, a monkeypox virus, a vaccinia virus, a variola virus, a rabbitpox virus, a raccoon poxvirus, a skunkpox virus, a Taterapox virus, an Uasin Gishu disease virus, and a volepox virus;

(iii) transfecting the overlapping DNA fragments derived from the overlapping chemically synthesized DNA fragments of step (i) and the ligated terminal hairpin loops of step (ii) into helper virus-infected cells;

(iv) culturing said cells to produce a mixture of helper virus and scOPV particles in said cells; and

(v) plating the mixture on OPV-specific host cells to recover the scOPV.

2. The method of claim 1 , wherein the helper virus is a leporipox virus.

3. The method of claim 2 , wherein the leporipox virus is selected from the group consisting of: Shope fibroma virus (SFV), hare fibroma virus, squirrel fibroma virus, and myxoma virus.

4. The method of claim 1 , wherein the helper virus is a fowlpox virus or a psoralen-inactivated helper virus.

5. The method of claim 1 , wherein the helper virus-infected cells are BGMK cells.

6. The method of claim 1 , wherein the OPV-specific host cells are BSC-40 cells.

7. A method of producing a synthetic chimeric horsepox virus (scHPXV) comprising the steps of:

(i) chemically synthesizing overlapping DNA fragments that correspond to substantially all of a wild type HPXV genome;

(ii) chemically synthesizing terminal hairpin loops independently derived from an orthopoxvirus and ligating the terminal hairpin loops derived from the chemically synthesized hairpin loops to the left and right termini of the overlapping DNA fragments derived from the chemically synthesized DNA of step (i), and wherein the orthopoxvirus is selected from the group consisting of a camelpox virus, a cowpox virus, an ectromelia virus, a horsepox virus, a monkeypox virus, a vaccinia virus, a variola virus, a rabbitpox virus, a raccoon poxvirus, a skunkpox virus, a Taterapox virus, an Uasin Gishu disease virus, and a volepox virus;

(iii) transfecting the overlapping DNA fragments derived from the overlapping chemically synthesized DNA fragments of step (i) and the ligated terminal hairpin loops of step (ii) into helper virus-infected cells;

(iv) culturing said cells to produce a mixture of helper virus and scHPXV particles in said cells; and

(v) plating the mixture on HPXV-specific host cells to recover the scHPXV.

8. The method of claim 7 , wherein the helper virus is a leporipox virus.

9. The method of claim 8 , wherein the leporipox virus is selected from the group consisting of: Shope fibroma virus (SFV), hare fibroma virus, squirrel fibroma virus, and myxoma virus.

10. The method of claim 7 , wherein the helper virus is a fowlpox virus or a psoralen-inactivated helper virus.

11. The method of claim 7 , wherein the helper virus-infected cells are BGMK cells.

12. The method of claim 7 , wherein the HPXV-specific host cells are BSC-40 cells.

13. The method of claim 9 , wherein the helper virus is a Shope fibroma virus (SFV).

14. The method of claim 13 , wherein the SFV-infected cells are BGMK cells.

15. The method of claim 13 , wherein the HPXV-specific host cells are BSC-40 cells.

16. A method of (i) triggering or boosting an immune response against one or more of a variola virus, a vaccinia virus, or a monkeypox virus, (ii) immunizing a subject to protect said subject from one or more of a variola virus infection, a vaccinia virus infection, or a monkeypox virus infection, or (iii) treating one or more of a variola virus infection, a vaccinia virus infection, or a monkeypox virus infection comprising administering to a subject in need thereof a composition comprising a synthetic chimeric orthopoxvirus (scOPV) of claim 1 .

17. A method of (i) triggering or boosting an immune response against one or more of a variola virus, a vaccinia virus, or a monkeypox virus, (ii) immunizing a subject to protect said subject from one or more of a variola virus infection, a vaccinia virus infection, or a monkeypox virus infection, or (iii) treating one or more of a variola virus infection, a vaccinia virus infection, or a monkeypox virus infection comprising administering to a subject in need thereof a composition comprising a synthetic chimeric horsepox virus (scHPXV) of claim 7 .

18. The method of claim 7 , wherein the overlapping DNA fragments comprise:

(i) nucleotide sequences that are at least 90% identical to the sequences of SEQ ID NOs: 1-10;

(ii) nucleotide sequences that are at least 95% identical to the sequences of SEQ ID NOs: 1-10; or

(iii) nucleotide sequences that consist of the sequences of SEQ ID NOs: 1-10.

19. The method of claim 16 , wherein the subject is a human.

20. The method of claim 17 , wherein the subject is a human.

Assignments (4)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Nov 21, 2024
From: TONIX PHARMACEUTICALS HOLDING CORP.
To: TONIX PHARMA LIMITED
Reel/Frame 069356/0314 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Jun 29, 2022
From: THE GOVERNORS OF THE UNIVERSITY OF ALBERTA
To: TONIX PHARMA LIMITED
Reel/Frame 060350/0620 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Jun 29, 2022
From: EVANS, DAVID; NOYCE, RYAN
To: THE GOVERNORS OF THE UNIVERSITY OF ALBERTA
Reel/Frame 060350/0666 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded May 27, 2022
From: LEDERMAN, SETH
To: TONIX PHARMACEUTICALS HOLDING CORP.
Reel/Frame 060042/0801 →
Continuity (4)
Division 15802189 · Nov 2, 2017
Provisional Application 62434794 · Dec 15, 2016
Provisional Application 62416577 · Nov 2, 2016
Related Publication 20230002741A1 · Jan 5, 2023
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Cited By (1)
US 12,529,036