Vaccines for the Treatment and Prevention of Zoonotic Infections

Abstract

The invention relates to compositions comprising a nucleic acid that encodes a peptide or a peptide that induces an immune response in an animal or a mammal that is protective against infection by one or more pathogens. In addition, the invention relates to vaccines comprising compositions and to method for treating and preventing an infection in animals and mammals such as humans.

Claims

1. An immunogenic composition comprising a viral or bacterial antigen of a pathogen containing an epitope that, upon administration to a human, a mammal or an animal, generates an immune response to the pathogen.

2. The composition of claim 1, wherein the antigen is obtained or derived from an HA protein, an NA protein, an M1 protein, an M2 protein, an M2e protein of an influenza virus, and/or a fragment, derivative, or modification thereof.

3. The composition of claim 1, wherein the antigen contains a sequence of any one or more of SEQ ID NOs. 1-106, any one or more of SEQ ID NOs 107-115, any one of more of SEQ ID NOs 116-131, or a combination thereof.

4. The composition of claim 1, further comprising T cell stimulating epitope obtained or derived from tetanus toxin, tetanus toxin heavy chain proteins, diphtheria toxoid, CRM, recombinant CRM, tetanus toxoid, Pseudomonas exoprotein A, Pseudomonas aeruginosa toxoid, Bordetella pertusis toxoid, Clostridium perfringens toxoid, Escherichia coli heat-labile toxin B subunit, Neisseria meningitidis outer membrane complex, Hemophilus influenzae protein D, Flagellin Fli C, Horseshoe crab Haemocyanin, and/or a fragment, derivative, or modification thereof.

5. The composition of claim 1, wherein the T cell stimulating epitope is at an N-terminus or a C-terminus of the antigen.

6. The composition of claim 1, which comprises multiple influenza virus epitopes and/or multiple T cell stimulating epitope.

7. The composition of claim 1, further comprising an adjuvant.

8. The composition of claim 7, wherein the adjuvant comprises Freund's adjuvant, ALFQ, ALFQA, ALFA, AS01, AS01b, a liposome adjuvant, saponin, lipid A, squalene, and/or modifications, derivatives and combinations thereof.

9. The composition of claim 1, which is a vaccine that treats or prevents a viral infection.

10. The composition of claim 1, which is a vaccine that treats or prevents a bacterial infection.

11. A method to treat or prevent an infection by the pathogen by administering the composition of claim 1 to a collection of animals suspected of being or determined to be infected with the pathogen.

12. The method of claim 11, wherein the composition produces a systemic and/or mucosal immune response by the animal.

13. The method of claim 11, wherein administration is to a water or food supply or as an aerosol.

14. The method of claim 11, wherein administration is oral, sub-cutaneous, intra-muscular, or intra-nasal.

15. A method to treat or prevent an infection by the pathogen by administering the composition of claim 1 to a human suspected of being or determined to be infected with the pathogen.

16. The method of claim 15, wherein the composition produces a systemic and/or mucosal immune response by the animal.

17. The method of claim 15, wherein administration is oral, sub-cutaneous, intra-muscular, or intra-nasal.

18. An immunogenic composition comprising a viral or bacterial nucleic acid that encodes an antigen of a pathogen containing an epitope that, upon administration to a human, a mammal or an animal, generates an immune repose to the pathogen.

19. The composition of claim 18, wherein the nucleic acid comprises DNA.

20. The composition of claim 18, wherein the nucleic acid comprises RNA.

Description

EXAMPLES

Example 1 Peptides and Sequences

[0070] The following is a list of exemplary peptide sequences. These sequences include composite sequences as well as sequences of interest that can be combined to form composite sequences:

Influenza Virus

[0071]

TABLE-US-00001 SEQ ID NO 1 DWSGYSGSFVQHPELTGLD (N1 sequence; H1 N5) SEQ ID NO 2 ETPIRNE (M2e epitope) SEQ ID NO 3 FVIREPFISCSHLEC SEQ ID NO 4 GNFIAP (HA epitope; Pep 1) SEQ ID NO 5 GNLIAP (HA epitope; Pep 2) SEQ ID NO 6 GNLFIAP (composite sequence of SEQ ID NOs 4 and 5; Pep 3) SEQ ID NO 7 GNLIFAP (composite sequence of SEQ ID NOs 4 and 5) SEQ ID NO 8 HYEECSCY (NA epitope; Pep 10) SEQ ID NO 9 LLTEVETPIR (highly conserved region M1/M2e) SEQ ID NO 10 LLTEVETPIRN (highly conserved region M1/M2e) SEQ ID NO 11 LLTEVETPIRNE (highly conserved region M1/M2e) SEQ ID NO 12 DWSGYSGSFVQHPELTGL (N1 sequence; H1 N5) SEQ ID NO 13 EVETPIRNE (highly conserved region M1/M2e) SEQ ID NO 14 FLLPEDETPIRNEWGLLTDDETPIRYIKANSKFIGITE SEQ ID NO 15 GNLFIAPGNLFIAPHYEECSCYHYEECSCYQYIKANSKFIGITEHYEECSCYTPIRNETPIRNE SEQ ID NO 16 GNLFIAPGNLFIAPQYIKANSKFIGITEGNLFIAP (composite of SEQ ID NO 6, SEQ ID NO 6, SEQ ID NO 61, and SEQ ID NO 6) SEQ ID NO 17 HYEECSCYDWSGYSGSFVQHPELTGLHYEECSCYQYIKANSKFIGITE SEQ ID NO 18 ITGFAPFSKDNSIRLSAGGDIWVTREPYVSCDP SEQ ID NO 19 IWGIHHP (HA epitope) SEQ ID NO 20 IWGVHHP (HA epitope) SEQ ID NO 21 IWGVIHHP (composite of SEQ ID NOs. 19 and 20) SEQ ID NO 22 IWGIVHHP (composite of SEQ ID NOs. 19 and 20) SEQ ID NO 23 KSCINRCFYVELIRGR (N3 conserved epitope) SEQ ID NO 24 LLTEVETPIRNESLLTEVETPIRNEWG (M2e epitope) SEQ ID NO 25 LLTEVETPIRNEW (M2e epitope) SEQ ID NO 26 LLTEVETPIRNEWG (M2e epitope) SEQ ID NO 27 LTEVETPIRNE (M2e epitope) SEQ ID NO 28 LTEVETPIRNEW (M2e epitope) SEQ ID NO 29 LTEVETPIRNEWG (M2e epitope) SEQ ID NO 30 MSLLTEVET (M2e epitope) SEQ ID NO 31 MSLLTEVETP (M2e epitope) SEQ ID NO 32 MSLLTEVETPI (M2e epitope) SEQ ID NO 33 MSLLTEVETPIR (M2e epitope) SEQ ID NO 34 MSLLTEVETPIRN (M2e epitope) SEQ ID NO 35 MSLLTEVETPIRNE (M2e epitopes) SEQ ID NO 36 MSLLTEVETPIRNETPIRNE (M2e epitope) SEQ ID NO 37 MSLLTEVETPIRNEW (M2e epitope) SEQ ID NO 38 MSLLTEVETPIRNEWG (M2e epitope) SEQ ID NO 39 MSLLTEVETPIRNEWGCRCNDSSD (M2e epitope) SEQ ID NO 40 SLLTEVET (M2e epitope) SEQ ID NO 41 SLLTEVETPIR (M2e epitope) SEQ ID NO 42 SLLTEVETPIRNE (M2e epitope) SEQ ID NO 43 SLLTEVETPIRNEW (M2e epitope) SEQ ID NO 44 SLLTEVETPIRNEWG (M2e epitope) SEQ ID NO 45 SLLTEVETPIRNEWGTPIRNE (M2e epitope) SEQ ID NO 46 SLLTEVETPIRNEWGTPIRNETPIRNE (M2e epitope) SEQ ID NO 47 SLLTEVETPIRNEWGTPIRNETPIRNETPIRNE (M2e epitopes) SEQ ID NO 48 SLLTEVETPIRNEWGLLTEVETPIR (M1/M2e conserved region) SEQ ID NO 49 TEVETPIRNE (M2e epitope) SEQ ID NO 50 TPIRNE (M2e epitope) SEQ ID NO 51 VETPIRNE (M2e epitope) SEQ ID NO 52 VTREPYVSCDPKSCINRCFYVELIRGRVTREPYVSCDPWYIKANSKFIGITE SEQ ID NO 53 WGIHHP (HA conserved region; Pep 5) SEQ ID NO 54 WGVHHP (HA conserved region; Pep 4) SEQ ID NO 55 WGVIHHP (composite of SEQ ID NOs 53 and 54; Pep 6) SEQ ID NO 56 WGIVHHP (composite of SEQ ID NOs 53 and 54; Pep 7) SEQ ID NO 57 YIWGIHHP (HA conserved region) SEQ ID NO 58 YIWGVHHP (HA conserved region) SEQ ID NO 59 YIWGVIHHP (composite of SEQ ID NOs 57 and 58) SEQ ID NO 60 YIWGIVHHP (composite of SEQ ID NOs 57 and 58) SEQ ID NO 61 QYIKANSKFIGITE (T cell epitope) SEQ ID NO 62 PIRNEWGCRCNDSSD (M2e epitope) SEQ ID NO 63 GNLFIAPWGVIHHPHYEECSCY (underlined sequences are epitopes HA {composite} (SEQ ID NO 6) and NA (SEQ ID NO 8), respectively, with middle as SEQ ID NO 55, of Influenza A; Pep 11) SEQ ID NO 64 CAGAGNFIAP SEQ ID NO 65 CAGAGNLIAP SEQ ID NO 66 CAGAGNLFIAP SEQ ID NO 67 CAGAWGVHHP SEQ ID NO 68 CAGAWGIHHP SEQ ID NO 69 CAGAWGVIHHP SEQ ID NO 70 CAGAWGIVHHP SEQ ID NO 71 GNLIAPWGVIHHP SEQ ID NO 72 CAGAGNLIAPWGVIHHP SEQ ID NO 73 GNLFIAPWGVIHHP SEQ ID NO 74 CAGAGNLFIAPWGVIHHP SEQ ID NO 75 HYEECSCY SEQ ID NO 76 CAGAHYEECSCY SEQ ID NO 77 CAGAGNLFIAPWGVIHHPHYEECSCY SEQ ID NO 78 GNLFIAPWGVIHHPGNLFIAPWGVIHHP SEQ ID NO 79 CAGAGNLFIAPWGVIHHPGNLFIAPWGVIHHP SEQ ID NO 80 HYEECSCYGNLFIAPWGVIHHP SEQ ID NO 81 GNLFIAPHYEECSCYWGVIHHP SEQ ID NO 82 SLLTEVETPIRNEWGLLTEVETPIRQYIKANSKFIGITE SEQ ID NO 83 GNLFIAPGNLFIAPQYIKANSKFIGITEGNLFIAP SEQ ID NO 84 HYEECSCYDWSGYSGSFVQHPELTGLHYEECSCYQYIKANSKFIGITE SEQ ID NO 85 VTREPYVSCDPKSCINRCFYVELIRGRVTREPYVSCDPQYIKANSKFIGITE SEQ ID NO 86 DWSGYSGSFVQHPELTGL SEQ ID NO 87 ITGFAPFSKDNSIRLSAGGDIWVTREPYVSCDP SEQ ID NO 88 KSCINRCFYVELIRGR SEQ ID NO 89 GNLFIAPRYAFA SEQ ID NO 90 CAGAGNLFIAPRYAFA SEQ ID NO 91 GNLVVPRYAFA SEQ ID NO 92 CAGAGNLVVPRYAFA SEQ ID NO 93 GNLIAPRYAFA SEQ ID NO 94 CAGAGNLIAPRYAFA SEQ ID NO 95 GNLVVP SEQ ID NO 96 CAGAGNLVVP SEQ ID NO 97 FVIREPFISCSHLEC SEQ ID NO 98 CAGAFVIREPFISCSHLEC SEQ ID NO 99 GNLFIAPWGVIHHPHYEECSCY (Pep 11) SEQ ID NO 100 GNLFIAPWGVIHHPHYEECSCYQYIKANSKFIGITE (Pep 11 with C terminal T cell epitope = Pep 63) SEQ ID NO 101 QYIKANSKFIGITEGNLFIAPWGVIHHPHYEECSCY (Pep 11 with N terminal T cell epitope = Pep 64) SEQ ID NO 102 HVEECSY (N1 and N2) SEQ ID NO 103 WFIHHP (H5) SEQ ID NO 104 DLWSYNAELLV (stem peptide) SEQ ID NO 105 DIWTYNAELLV (stem peptide) SEQ ID NO 106 HXXXW-matrix peptide common to Flu A and B that constitutes the main functional element of the M2 channel

Coronavirus

[0072]

TABLE-US-00002 SEQ ID NO 107 YPKCDRA = RNA Polymerase region SEQ ID NO 108 WDYPKCDRA = RNA Polymerase region SEQ ID NO 109 SLDQINVTFLDLEYEMKKLEESY w/ QYIKANSKFIGITE = spike protein w/ tetanus toxoid T cell epitope Five coronavirus composite sequences using conserved epitopes. SEQ ID NO 110 SLDQINVTFLDLEYEMKKLEESY (coronavirus spike protein conserved epitope (SP)) SEQ ID NO 111 SLDQINVTFLDLEYEMKKLEESYQYIKANSKFIGITE (tetanus toxoid T cell epitope + SP) SEQ ID NO 112 WDYPKCDRA (polymerase conserved epitope (POL)) SEQ ID NO 113 WDYPKCDRAQYIKANSKFIGITE (POL + tetanus T cell epitope) SEQ ID NO 114 WDYPKCDRASLDQINVTFLDLEYEMKKLEESYQYIKANSKFIGITE (Cor POL + SP + Tet) SEQ ID NO 115 WDYPKCDRATEVETPIRNEHYEECSCYQYIKANSKFIGITE Cor POL. Flu M2e. Flu NA. Tetanus T cell (One coronavirus conserved epitope and two Flu conserved epitopes that is a broader pandemic vaccine)

Coronavirus Peptides and Composite Coronavirus/Influenza Peptides

[0073]

TABLE-US-00003 SEQ ID NO 116 SLDQINVTFLDLEYEMKKLEESYQYIKANSKFIGITE (spike protein epitope with T-cell  stimulating epitope) SEQ ID NO 117 WDYPKCDRA (corona conserved seq-polymerase) neutralizing Ab SEQ ID NO 118 YPKCDRA (corona conserved seq-polymerase) SEQ ID NO 119 ARDLICAQ (highly conserved cor seq-spike attachment same is all three-Cor MERS SARS) SEQ ID NO 120 KWPWYIWLGFIAGL (highly conserved cor seq-spike attachment) SEQ ID NO 121 ENQKLIAN (highly conserved cor seq-spike attachment) SEQ ID NO 122 ARDLICAQKWPWYIWLGFIAGLENQKLIAN (combination of conserved seqs w/o T cell epitope) SEQ ID NO 123 ENQKLIANARDLICAQ (combination of conserved seqs w/o T cell epitope) SEQ ID NO 124 WDYPKCDRAENQKLIANARDLICAQ (combination of conserved seqs w/o T cell epitope) SEQ ID NO 125 WDYPKCDRAENQKLIANKWPWYIWLGFIAGL (combination of conserved seqs w/o T cell epitope) SEQ ID NO 126 ARDLICAQENQKLIANWDYPKCDRA (combinations of cor conserved seqs w/ T cell epitope) SEQ ID NO 127 KWPWYIWLGFIAGLWDYPKCDRAQYIKANSKFIGITEARDLICAQEN QKLIANWDYPKCDRA (combination of cor conserved seqs w/ T cell epitope) SEQ ID NO 128 ARDLICAQENQKLIANQYIKANSKFIGITE ARDLICAQENQKLIAN WDYPKCDRA (combination of cor conserved seqs w/ T cell epitope) SEQ ID NO 129 WDYPKCDRATEVETPIRNE ARDLICAQENQKLIANWDYPKCDRAQYIKANSKFIGITE (combination of cor plus Influenza conserved seqs w/ T cell epitope) Just bold = Cor Italicized = m2e Underlined = Flu Bold and underlined—T-cell SEQ ID NO 130 HYEECSCYWDYPKCDRAVETPIRNE (combination of cor plus Influenza conserved seqs w/ T cell epitope) SEQ ID NO 131 ENQKLIANTEVETPIRNE (combination of cor plus Influenza conserved seqs w/ T cell epitope)

Example 2 Induction of Neutralizing Antibodies with Composite Peptides

[0074] ICR mice and cotton rats were immunized with 1 μg of conjugated or unconjugated composite influenza peptide vaccine (influenza HA, NA and M2e composite peptides with a T-cell epitope) formulated with ALFQ by intramuscular, or intradermal routes (cotton rats were given both intramuscular, or intradermal injections). Both routes of administration induced serum IgG antibodies that bound to groups 1 and 2 influenza viruses (Flu A California H1N1/pdm09 and Flu A Hong Kong H3N2/4801/2014). In addition, 1 μg of composite influenza vaccine formulated in ALFQ induced neutralizing antibodies against both influenza viruses given by intradermal and intramuscular routes. These data demonstrate that composite influenza peptide vaccines formulated in ALFQ induced a strong immune response at a very low dose without conjugation to a carrier and when administered by different routes of immunization. This provides an advantage in efficiency of manufacturing and decreased cost of production. Low dose intradermal administration also decreases vaccine costs for mass global immunization of humans and for immunizing mammals such as humans or animals such as birds and pigs.

[0075] Other embodiments and uses of the invention will be apparent to those skilled in the art from consideration of the specification and practice of the invention disclosed herein. All references cited herein, including all publications, U.S. and foreign patents and patent applications, are specifically and entirely incorporated by reference. The term comprising, wherever used, is intended to include the terms consisting and consisting essentially of. Furthermore, the terms comprising, including, and containing are not intended to be limiting. It is intended that the specification and examples be considered exemplary only with the true scope and spirit of the invention indicated by the following claims.