SARS-CoV-2 subunit vaccine and microneedle array delivery system

Patent metadata

Field	Value
Publication number	US-11213482-B1
Application number	US-202017009121-A
Country	US
Kind code	B1
Filing date	Sep 1, 2020
Priority date	Mar 5, 2020
Publication date	Jan 4, 2022
Grant date	Jan 4, 2022

Abstract

A recombinant coronavirus vaccine is provided. Methods of making and delivering the coronavirus vaccine also are provided. A microneedle array is provided, along with methods of making and using the microneedle array.

First claim

What is claimed is:

1. A pharmaceutical composition comprising (a) a polypeptide having the sequence of amino acids 1-661 of SEQ ID NO: 2 or a derivative thereof having at least 95% sequence identity with amino acids 1-661 of SEQ ID NO: 2, and (b) a TLR3 agonist, a TLR 4 agonist, a TLR 5 agonist, a TLR 7/8 agonist, a TLR 9 agonist, a Stimulator of Interferon Genes (STING) pathway agonist, a stimulatory neuroimmune mediator, a neurokinin 1 (NK1) receptor agonist, a saponin related adjuvant, a purinoergic receptor agonist, or an oil-in-water emulsion adjuvant.

2. The composition of claim 1 , wherein said composition comprises said polypeptide having the sequence of amino acids 1-661 of SEQ ID NO: 2.

3. The composition of claim 2 , wherein the polypeptide is a fusion protein comprising a multimerization domain, a Toll-like receptor agonist domain, or a SARS-CoV-2 nucleoprotein amino acid sequence having the sequence of amino acids 664-1,082 of SEQ ID NO:16 or a derivative thereof having at least 95% sequence identity with amino acids 664-1,082 of SEQ ID NO: 16.

4. The composition of claim 1 , wherein said composition comprises said derivative having at least 95% sequence identity with amino acids 1-661 of SEQ ID NO:2.

5. The composition of claim 1 , wherein said composition comprises said oil-in-water emulsion adjuvant.

6. The composition of claim 1 , wherein said derivative has at least 99% sequence identity with amino acids 1-661 of SEQ ID NO:2.

7. A microneedle array comprising: a backing layer; and a plurality of microneedles, with a polypeptide having (a) the sequence of SEQ ID NO: 2, (b) amino acids 1-661 of SEQ ID NO: 2, (c) a sequence at least 95% sequence identity with amino acids 1-661 of SEQ ID NO: 2, or (d) a sequence having at least 95% sequence identity with SEQ ID NO: 2, incorporated into or onto each of said plurality of microneedles.

8. The microneedle array of claim 7 , wherein each of said plurality of microneedles comprises a matrix of a dissolvable or bioerodible, biocompatible material extending from said backing layer.

9. The microneedle array of claim 8 , wherein each of said plurality of microneedles comprises a plurality of layers of one or more dissolvable or bioerodible, biocompatible material.

10. The microneedle array of claim 8 , wherein each of said plurality of microneedles comprises carboxymethyl cellulose, trehalose, polyvinylpyrrolidone, maltodextrin, silk, hyaluronic acid, poly(lactic-co-glycolic acid), poly(lactic acid), poly(vinyl alcohol), polyethylene glycol, or a combination of any two or more of the preceding.

11. The microneedle array of claim 8 , wherein each of said plurality of microneedles comprises carboxymethyl cellulose.

12. The microneedle array of claim 11 , wherein each of said plurality of microneedles further comprises trehalose.

13. The microneedle array of claim 8 , wherein each of said plurality of microneedles has a shape that comprises a first cross-sectional dimension at a head portion distal to the backing layer, a second cross-sectional dimension at a stem portion proximal to the backing layer, and a third cross-sectional dimension at an intermediate portion located between said head portion and said stem portion having a cross-sectional dimension greater than the first and second cross-sectional dimensions.

14. The microneedle array of claim 8 , wherein each of said plurality of microneedles comprises a stem portion adjacent or proximal to the backing layer and a head portion attached to the stem portion distal to the backing layer, wherein each of the plurality of microneedles has a barbed or undercut profile in which the head portion having a cross section adjacent to the stem portion is larger than a cross section of the stem portion adjacent to the head portion.

15. The microneedle array of claim 8 , wherein each of said plurality of microneedles comprises a stem, a head, a filleted base, and at least one undercut feature.

16. A method of eliciting an immune response against SARS-CoV-2 in a patient, comprising administering to the patient an amount of the pharmaceutical composition of claim 1 effective to elicit an immune response against SARS-CoV-2 in a patient, thereby eliciting an immune response in the patient against SARS-CoV-2.

17. The method of claim 16 , wherein said composition comprises said polypeptide having the sequence of amino acids 1-661 of SEQ ID NO:2.

18. The method of claim 17 , wherein the patient is a human.

19. A method of forming a microneedle array comprising forming a plurality of microneedles having a filleted base attached to a backing, comprising incorporating into or onto each of said plurality of microneedles a polypeptide having (a) the sequence of SEQ ID NO: 2, (b) amino acids 1-661 of SEQ ID NO:2, (c) a sequence having at least 95% sequence identity with amino acids 1-661 of SEQ ID NO: 2, or (d) a sequence having at least 95% sequence identity with SEQ ID NO: 2.

20. The method of forming a microneedle array of claim 19 , wherein the polypeptide is incorporated into a first dissolvable or bioerodible material.

21. The method of forming a microneedle array of claim 20 , comprising:
forming or providing a production mold of a flexible material, the production mold comprising a plurality of cavities that are shaped to define a plurality of respective microneedles having a stem, a head, a filleted base, and at least one undercut feature;
delivering the first dissolvable or bioerodible material into at least a microneedle head portion defined by the respective cavities of the production mold, and prior to or during delivery of the first dissolvable or bioerodible material into at least the microneedle head portion, incorporating the polypeptide into the first dissolvable or bioerodible material to provide a biodegradable matrix;
delivering (a) the first dissolvable or bioerodible material, (b) one or more additional dissolvable or bioerodible materials, or (c) a combination thereof, into the cavity and forming a plurality of microneedles in the production mold that include the biodegradable matrix; and
removing the microneedles from the production mold by pulling the microneedles out of the mold,
wherein the flexible material of the production mold has sufficient elasticity to allow for the molded microneedle array to be removed from the production mold in a single pull, without damaging the integrity of the shape of the microneedles as defined by the mold.

22. The method of claim 21 , wherein the at least one undercut feature is directly below the microneedle head.

23. The method of claim 21 , wherein the stem is formed from the first dissolvable or bioerodible material.

24. The method of claim 21 , wherein the first dissolvable or bioerodible material comprises carboxymethyl cellulose, trehalose, polyvinylpyrrolidone, maltodextrin, silk, hyaluronic acid, poly(lactic-co-glycolic acid), poly(lactic acid), poly(vinyl alcohol), polyethylene glycol, or a combination of any two or more of the preceding.

25. The method of claim 21 , further comprising delivering a second dissolvable material into the production mold to form a dissolving layer at a portion of the stem.

26. The method of claim 25 , wherein the second dissolvable material of the dissolving layer is a material that dissolves more quickly than the biodegradable matrix that contains the polypeptide.

27. The method of claim 21 , wherein the mi

Assignees

• Univ Of Pittsburgh—Of The Commonwealth System Of Higher Education
• Univ Of Pittsburgh—Of The Commonwealth System Of Higher Educat

Inventors

• Gambotto Andrea
• Falo Jr Louis
• Kim Eun

Classifications

• C12N2770/20034

  Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein · CPC title

• C12N2710/10343

  viral genome or elements thereof as genetic vector · CPC title

• A61P31/14

  for RNA viruses · CPC title

• A61M2202/30

  Vaccines · CPC title

• A61M2037/0053

  Methods for producing microneedles · CPC title