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US-20260124297-A1 - NOVEL ADJUVANT FOR ANIMAL AND HUMAN VACCINES

US20260124297A1US 20260124297 A1US20260124297 A1US 20260124297A1US-20260124297-A1

Abstract

Quil-A chitosan spherical nanostructure complexes as well as methods of making and using such complexes are disclosed herein. Also provided are Quil-A chitosan spherical nanostrucutres loaded with one or more RNA, DNA, or protein payload molecules as well as methods of making and using such loaded complexes.

Inventors

  • Adel M. Talaat
  • Shaswath Chandrasekar

Assignees

  • WISCONSIN ALUMNI RESEARCH FOUNDATION

Dates

Publication Date
20260507
Application Date
20251222

Claims (20)

  1. 1 . A method of immunizing a subject against an antigen comprising the step of administering to the subject a vaccine formulation comprising a composition comprising disaggregated spherical nanostructures comprising Quil-A and chitosan and a payload, wherein the Quil-A and chitosan are present at a ratio between 1:15 weight:weight, and 1:100 weight:weight, and wherein the payload comprises the antigen or a polynucleotide encoding the antigen.
  2. 2 . The method of claim 1 , wherein the subject is selected from the group consisting of a human, a mouse, a rat, a cow, a horse, a pig, a goat, a sheep, a cat, a dog, or a bird.
  3. 3 . The method of claim 1 , wherein the antigen comprises a polypeptide.
  4. 4 . The method of claim 3 , wherein the polypeptide is derived from a virus, bacteria, or fungus.
  5. 5 . The method of claim 1 , wherein the polynucleotide encoding the antigen is a DNA molecule or an RNA molecule.
  6. 6 . The method of claim 5 , wherein the polynucleotide encoding the antigen is a DNA molecule and is operably liked to a promoter.
  7. 7 . The method of claim 1 , wherein the antigen is negatively charged.
  8. 8 . The method of claim 1 , wherein the Quil-A and chitosan are present at a ratio between 1:15 weight:weight, and 1:50 weight:weight.
  9. 9 . The method of claim 1 , wherein the Quil-A and chitosan are present at a ratio of about 1:20 weight:weight.
  10. 10 . The method of claim 1 , wherein administration comprises intranasal administration, transmucosal administration, subcutaneous administration, or intramuscular administration.
  11. 11 . A method of eliciting an immune response to an antigen in a subject, the method comprising administering to the subject a composition comprising disaggregated spherical nanostructures comprising Quil-A and chitosan and a payload, wherein the Quil-A and chitosan are present at a ratio between 1:15 weight:weight, and 1:100 weight:weight, and wherein the payload comprises the antigen or a polynucleotide encoding the antigen.
  12. 12 . The method of claim 11 , wherein the subject is selected from the group consisting of a human, a mouse, a rat, a cow, a horse, a pig, a goat, a sheep, a cat, a dog, or a bird.
  13. 13 . The method of claim 11 , wherein the antigen comprises a polypeptide.
  14. 14 . The method of claim 13 , wherein the polypeptide is derived from a virus, bacteria, or fungus.
  15. 15 . The method of claim 11 , wherein the polynucleotide encoding the antigen is a DNA molecule or an RNA molecule.
  16. 16 . The method of claim 15 , wherein the polynucleotide encoding the antigen is a DNA molecule and is operably liked to a promoter.
  17. 17 . The method of claim 11 , wherein the antigen is negatively charged.
  18. 18 . The method of claim 11 , wherein the Quil-A and chitosan are present at a ratio between 1:15 weight:weight, and 1:50 weight:weight.
  19. 19 . The method of claim 11 , wherein the Quil-A and chitosan are present at a ratio of about 1:20 weight:weight.
  20. 20 . The method of claim 11 , wherein administration comprises intranasal administration, transmucosal administration, subcutaneous administration, or intramuscular administration.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS This application is a divisional of U.S. application Ser. No. 18/450,908, filed Aug. 16, 2023, which is a divisional of U.S. application Ser. No. 16/900,070, filed Jun. 12, 2020, which claims priority to U.S. Provisional Application No. 62/860,640, filed Jun. 12, 2019, each of which are incorporated by reference herein in their entireties. STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH This invention was made with government support under 2016-67021-25042 awarded by the USDA/NIFA. The government has certain rights in the invention. REFERENCE TO A SEQUENCE LISTING SUBMITTED VIA PATENT CENTER The content of the xml file of the sequence listing named “960296_04450.xml” which is 64,839 bytes in size was created on Nov. 3, 2023 and electronically submitted via Patent Center is incorporated herein by reference in its entirety. BACKGROUND Vaccines have been hailed as one of the greatest achievements in public health during the past century. Vaccines have been a key factor for fighting infectious diseases that afflict humans and animals, with corresponding increases in human average life expectancy. The global eradication of Smallpox virus in humans and Rinderpest virus in animals, and the near eradication or successful prevention of other viral or bacterial infections, for example meningitis in children due to Hemophilus influenzae Type B, offer compelling examples. Adjuvants play a key role in the successful use of vaccines in human and animal medicines. However, only a handful of such adjuvants are approved for human and animal use. Needed in the art are additional vaccine adjuvant compositions for the improvement of human and animal medicines. SUMMARY OF THE INVENTION In a first aspect, provided herein is a composition comprising disaggregated spherical nanostructures comprising Quil-A and chitosan. The disaggregated spherical nanostructures may additionally comprising a payload molecule. In some embodiments, the payload molecule is selected from the group consisting of a DNA molecule, an RNA molecule, a polynucleotide, a protein, a polypeptide, a virus, a microbe, an attenuated virus, an attenuated microbe, a small molecule, an antibody, or a mixture thereof. In some embodiments, the payload molecule is negatively charged. In some embodiments, the payload is functionalized. In some embodiments, the payload molecule is an antigen specific for Mycobacterium avium subspecies paratuberculosis, Mycobacterium bovis, Mycobacterium tuberculosis, or Mycobacterium avium subspecies avium. In some embodiments, the chitosan is functionalized by treatment with 5-formyl-2-furan sulfonic acid and sodium borohydride such that the chitosan surface is negatively charged. In some embodiments, the spherical nanostructures are between about 5 nm and about 100 nm in diameter in the absence of a payload molecule. In a second aspect, provided herein is a vaccine formulation comprising an antigen, the disaggregated spherical nanostructures comprising Quil-A and chitosan described herein as an adjuvant, and a pharmaceutically acceptable carrier. In a third aspect, provided herein is a vaccine formulation comprising an antigen specific for Mycobacterium avium subspecies paratuberculosis, Mycobacterium bovis, Mycobacterium tuberculosis, or Mycobacterium avium subspecies avium and the disaggregated spherical nanostructures comprising Quil-A and chitosan described herein. In a forth aspect, provided herein is a method of forming a composition comprising Quil-A chitosan spherical nanostructures, comprising the steps of heating a first solution comprising Quil-A at about 55° C. for about 30 minutes; heating a second solution comprising chitosan at about 55° C. for about 30 minutes; mixing equal volumes of the first and second solution dropwise to form a combined solution; vortex mixing the combined solution for about 30 seconds to form a combined, vortexed solution; and incubating the combined vortexed solution whereby a composition comprising Quil-A chitosan spherical nanostructures is formed. In some embodiments, the combined vortexed solution is incubated at room temperature for about 1 hour. In some embodiments, the first solution additionally comprises a DNA antigen. In some embodiments, the combined vortexed solution is incubated at about 37 C with shaking at about 110 rpm for about 1 hour. In some embodiments, the first solution additionally comprises a protein antigen. In some embodiments, the first solution comprises about 0.002% Quil-A and the second solution comprises about 0.04% chitosan. In some embodiments, the first solution and the second solution each have a pH between 5.5 and 7.0. In a fifth aspect, provided herein is a composition comprising Quil-A chitosan spherical nanostructures produced by the methods described herein. In some embodiments the composition additionally comprises an antigen payload molecule. In a sixth aspect, provided herein is a method of immunizing a subject aga