AU2023200517B2

AU2023200517B2 - Consensus prostate antigens nucleic acid molecule encoding the same and vaccine and uses comprising the same

Info

Publication number: AU2023200517B2
Application number: AU2023200517A
Authority: AU
Inventors: Bernadette Ferraro; Mathura P. Ramanathan; Niranjan Y. Sardesai; David B. Weiner; Jian Yan
Original assignee: University of Pennsylvania Penn; Inovio Pharmaceuticals Inc
Current assignee: University of Pennsylvania Penn; Inovio Pharmaceuticals Inc
Priority date: 2010-11-12
Filing date: 2023-02-01
Publication date: 2025-09-04
Anticipated expiration: 2031-11-14
Also published as: AU2020203908B2; AU2015261742A1; AU2015261742B2; AU2020203908A1; AU2023200517A1; AU2018200542A1; AU2018200542B2

Abstract

#$%^&*AU2023200517B220250904.pdf##### ABSTRACT Provided herein are consensus amino acid sequences of prostate antigens that are capable of breaking tolerance in a targeted species, including PSA, PSMA, STEAP and PSCA antigens. Also provided are nucleic acid sequences that encode one or more consensus amino acid sequences of prostate antigens PSA, PSMA, STEAP and PSCA, as well as genetic constructs/vectors and vaccines expressing the sequences. Also provided herein are methods for generating an autoimmune response against prostate cancer cells by administering one or more of the vaccines, proteins, and/or nucleic acid sequences that are provided. ABSTRACT 2023200517 01 Feb 2023 Provided herein are consensus amino acid sequences of prostate antigens that are capable of breaking tolerance in a targeted species, including PSA, PSMA, STEAP and PSCA antigens. Also provided are nucleic acid sequences that encode one or more consensus amino acid sequences of prostate antigens PSA, PSMA, STEAP and PSCA, as well as genetic constructs/vectors and vaccines expressing the sequences. Also provided herein are methods for generating an autoimmune response against prostate cancer cells by administering one or more of the vaccines, proteins, and/or nucleic acid sequences that are provided.

Description

AUSTRALIA AUSTRALIA 2023200517 01 Feb

Patents Act Patents 1990 Act 1990

TheTrustees The Trusteesofofthe theUniversity Universityof of Pennsylvania Pennsylvania

Inovio Pharmaceuticals, Inovio Pharmaceuticals, Inc. Inc.

COMPLETESPECIFICATION COMPLETE SPECIFICATION STANDARD PATENT STANDARD PATENT

Invention Title: Invention Title:

Consensus prostate Consensus prostate antigens antigens nucleic nucleic acidacid molecule molecule encoding encoding the the same same and andand vaccine vaccine uses and uses

comprisingthe comprising the same same

-1-

CONSENSUS PROSTATEANTIGENS, CONSENSUS PROSTATE ANTIGENS,NUCLEIC NUCLEICACID ACIDMOLECULE MOLECULE ENCODING ENCODING THE SAME AND VACCINE AND USES COMPRISING THE SAME THE SAME AND VACCINE AND USES COMPRISING THE SAME

RELATEDAPPLICATION APPLICATIONDATA DATA 2023200517 01 Feb

RELATED This application This applicationisis aa divisional divisional application applicationofofAustralian AustralianApplication Application No. No. 2020203908, 2020203908,

whichitself which itself isisa adivisional application divisional of Australian application Application of Australian ApplicationNo. No. 2018200542, which 2018200542, which itself itself is is

a divisional application a application of ofAustralian AustralianApplication Application No.No. 2015261742 2015261742 pursuantpursuant to 79B to Section Section 79B of the Patents of Act1990 Patents Act 1990ofof Australian Australian Application Application No. No. 2011325893 2011325893 which corresponds which corresponds to to International Application International ApplicationNo. No.PCT/US2011/060592 filed 14 PCT/US2011/060592 filed 14 November November2011 2011 ininthe the Australiannational Australian nationalphase, phase,which which claims claims priority priority to Provisional to US US Provisional Application Application No. No. 61/413176 filed 61/413176 filed 12 12 November 2010and November 2010 andUSUSProvisional ProvisionalApplication ApplicationNo. No.61/417817 61/417817filed filed 29 November 29 November 2010, 2010, the complete the complete contents contents of are of which which are incorporated incorporated herein inherein in their entirety. their entirety.

FIELD FIELD OF OF THE THE INVENTION INVENTION Thepresent The presentinvention invention relatestotonucleic relates nucleic acid acid sequences sequences encoding encoding consensus consensus prostate prostate

proteins and proteins andfragments fragments thereof; thereof; to to improved improved prostate prostate cancer cancer vaccines, vaccines, improved improved methods methods for for inducingimmune inducing immune responses responses against against prostate prostate cancercancer cells,cells, improved improved methods methods for for prophylacticallyand/or prophylactically and/ortherapeutically therapeutically immunizing immunizing individuals individuals against against prostate prostate cancer. cancer.

BACKGROUND OFTHE BACKGROUND OF THE INVENTION INVENTION

Prostate cancer Prostate cancerisis an animportant importanttherapeutic therapeutic immune immune target. target. The development The development of an of an immune immune therapeutic therapeutic approach approach is complex, is complex, in immunogens in that that immunogens need need to be to be developed developed that are that are capableofofinducing capable inducingstrong strong immune immune responses responses including including preferably preferably CTL responses. CTL responses.

Thedirect The direct administration administrationofof nucleic nucleic acid acid sequences sequences to vaccinate to vaccinate against against animal animal and and human human diseases diseases hashas been been studied studied and much and much effort effort has focused has focused on effective on effective and efficient and efficient

meansofofnucleic means nucleicacid acid delivery delivery in in order order to to yield yield necessary necessary expression expression of the ofdesired antigens, the desired antigens, resulting immunogenic resulting immunogenic response response and ultimately and ultimately the success the success of thisof this technique. technique.

DNAvaccines DNA vaccineshave havemany many conceptual conceptual advantages advantages over over more more traditional vaccination traditional vaccination methods,such methods, such as as liveattenuated live attenuated viruses viruses andand recombinant recombinant protein-based protein-based vaccines. vaccines. DNA DNA vaccinesare vaccines aresafe, safe, stable, stable, easily easily produced, produced,and and well well tolerated tolerated in in humans humans with with preclinical preclinical trials trials

indicating little indicating little evidence of plasmid evidence of plasmidintegration integration[Martin,

[Martin,T.,T.,etetal., al., Plasmid PlasmidDNADNA malaria malaria

vaccine: the vaccine: the potential potential for for genomic genomic integration integration after after intramuscular intramuscular injection. injection. Hum Hum Gene Ther, Gene Ther, 1999. 10(5): 1999. 10(5): p. p. 759-68; 759-68;Nichols, Nichols,W.W., W.W., et al., et al., Potential Potential DNADNA vaccine vaccine integration integration into into host host cell genome. cell Ann genome. Ann NY N Y Acad Acad Sci, 772: Sci, 1995. 1995.p.772: p. In 30-9]. 30-9]. In addition, addition, DNAare DNA vaccines vaccines well are well suited for suited for repeated administrationduedue repeated administration to to the the factthat fact thatefficacy efficacyofof thevaccine the vaccine is is notnot influenced influenced

-2-

Feb 2023 by pre-existing by pre-existingantibody antibodytiters titerstotothe thevector vector[Chattergoon,

[Chattergoon,M.,M., J. Boyer, J. Boyer, and and D.B. D.B. Weiner, Weiner,

Geneticimmunization: Genetic immunization: a new a new eravaccines era in in vaccines and immune and immune therapeutics. therapeutics. FASEB J, FASEB 1997. J, 1997. 11(10): p. 11(10): p. 753-63]. 753-63]. However, However, one one majormajor obstacle obstacle forclinical for the the clinical adoption adoption of DNA of DNA vaccines vaccines has been has beena adecrease decreaseininthe theplatform's platform's immunogenicity immunogenicity when to when moving moving larger to larger[Liu, animals animals [Liu, 2023200517 01 M.A.and M.A. andJ.B. J.B.Ulmer, Ulmer, Human Human clinical clinical trialstrials of plasmid of plasmid DNA vaccines. DNA vaccines. Adv Adv Genet, Genet, 2005. 55: 2005. 55: p. 25-40]. p. 25-40]. Recent Recent technological technologicaladvances advances ininthe engineering the of of engineering DNADNAvaccine vaccineimmunogen, immunogen,

such has such hascodon codon optimization, optimization, RNARNA optimization optimization and theand the addition addition of immunoglobulin of immunoglobulin leader leader sequenceshave sequences have improved improved expression expression and immunogenicity and immunogenicity of DNA of DNA vaccines vaccines

[Andre, [Andre, S., et al., S., et al., Increasedimmune Increased immune response response elicited elicited by vaccination by DNA DNA vaccination with a synthetic with a synthetic gp120 gp120 sequence sequence with optimized with optimizedcodon codon usage. usage. J Virol, J Virol, 1998. 1998. 72(2): 72(2): p. 1497-503; p. 1497-503; Deml, Deml, L., etMultiple L., et al., al., Multiple effects of effects of codon usageoptimization codon usage optimization on expression on expression and immunogenicity and immunogenicity of DNA of DNA candidate candidate vaccinesencoding vaccines encodingthethe human human immunodeficiency immunodeficiency virus virus type type 1 Gag 1 GagJprotein. protein. J Virol, Virol, 2001. 2001. 75(22): p. 75(22): p. 10991-1001; 10991-1001; Laddy, Laddy, D.J., D.J., et al.,Immunogenicity et al., Immunogenicity of novel of novel consensus-based consensus-based DNA DNA vaccinesagainst vaccines againstavian avianinfluenza. influenza.Vaccine, Vaccine, 2007. 2007. 25(16): 25(16): p. 2984-9; p. 2984-9; Frelin, Frelin, L.,al., L., et et al., CodonCodon optimizationand optimization andmRNA mRNA amplification amplification effectively effectively enhances enhances the immunogenicity the immunogenicity of the of the hepatitis C hepatitis virus nonstructural C virus nonstructural3/4A 3/4A gene. gene. Gene Gene Ther, Ther, 2004.2004. 11(6):11(6): p. 522-33]. p. 522-33].

Recenttechnological Recent technologicaladvances advances in plasmid in plasmid delivery delivery systems systems have improved have improved expressionexpression

and immunogenicity and immunogenicity of DNA of DNA vaccines vaccines including including technologies technologies such as electroporation such as electroporation [Hirao, [Hirao, L.A., et L.A., et al., al., Intradermal/subcutaneous immunization Intradermal/subcutaneous immunization by electroporation by electroporation improves improves plasmid plasmid vaccinedelivery vaccine deliveryand andpotency potency in pigs in pigs and and rhesus rhesus macaques. macaques. Vaccine, Vaccine, 2008. p.26(3): 2008. 26(3): 440-8;p. 440-8; Luckay,A., Luckay, A.,etetal., al., Effect Effect of of plasmid plasmidDNA DNA vaccine vaccine design design and inand in electroporation vivo vivo electroporation on the on the resulting vaccine-specific resulting vaccine-specificimmune immune responses responses in rhesus in rhesus macaques. macaques. J Virol, J Virol, 2007. 81(10): 2007. 81(10): p. p. 5257-69;Ahlen, 5257-69; Ahlen,G.,G.,et etal., al., InInvivo vivoelectroporation electroporationenhances enhances the the immunogenicity immunogenicity of hepatitis of hepatitis

C virus C virus nonstructural nonstructural3/4A 3/4ADNADNA by increased by increased localuptake, local DNA DNA protein uptake,expression, protein expression, inflammation,andand inflammation, infiltrationofofCD3+ infiltration CD3+ T cells. T cells. J Immunol, J Immunol, 2007. 2007. 179(7):179(7): p. 4741-53]. p. 4741-53].

In addition, In studies have addition, studies havesuggested suggestedthat thatthetheuseuseof of consensus consensus immunogens immunogens can can be able be able to increase to the breadth increase the breadthofofthe thecellular cellular immune immune response response as compared as compared to native to native antigens antigens alone alone

[Yan, J.,

[Yan, J., et et al., al.,Enhanced cellular immune Enhanced cellular immune responses responses elicited elicited byengineered by an an engineered HIV-1 HIV-1 subtype subtype

B consensus-based B consensus-based envelope envelope DNA vaccine. DNA vaccine. Mol Mol Ther, Ther, 2007. 2007. 15(2): 15(2): p.Rolland, p. 411-21; 411-21;M.,Rolland, et M., et al., Reconstruction al., andfunction Reconstruction and functionof of ancestral ancestral center-of-tree center-of-tree human human immunodeficiency immunodeficiency virus virus type 11 proteins. type proteins. JJ Virol, Virol, 2007. 2007. 81(16): 81(16):p.p.8507-14]. 8507-14].However, However, it is itrecognized is recognized that breaking that breaking

immune immune tolerance tolerance forfor cancer cancer antigens antigens and generating and generating autoimmunity autoimmunity is aobstacle is a major major obstacle for for cancer vaccines. cancer vaccines.

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Feb 2023 Therestill There remains aa need still remains needfor fornucleic nucleicacid acidconstructs constructs that that encode encode prostate prostate cancer cancer

antigens and antigens andfor forcompositions compositions useful useful to induce to induce immune immune responses responses prostateprostate against against cancer cancer antigens and antigens andthus thusbreak breakimmune immune tolerance. tolerance. There There remainsremains a need a need for for effective effective prophylactic prophylactic

and therapeutic and therapeuticvaccines vaccinesagainst against prostate prostate cancer cancer thatthat areare economical economical and effective. and effective.

2023200517 01

SUMMARY OF SUMMARY OF THE THE PREFERRED PREFERRED EMBODIMENTS EMBODIMENTS

Aspects Aspects ofofthe thepresent presentinvention invention include include nucleic nucleic acidacid molecules molecules comprising comprising a coding a coding

sequenceencoding sequence encoding one one or more or more proteins proteins selected selected from from the the comprising: group group comprising: a) SEQ IDa) SEQ ID NO:2; NO:2; a aprotein proteinthat thatisis 98% 98% homologous homologous to SEQtoIDSEQ IDprovided NO:2, NO:2, provided amino amino acids acids 69, 78, 80, 69, 78, 80,

82, 102, 82, 102, 110, 110, 137, 137,139, 139,165, 165,189, 189,203, 203, 220, 220, 232232 and and 248SEQofIDSEQ 248 of NO:2ID NO:2 are are conserved; conserved; or or an immunogenic an fragmentofofSEQ immunogenic fragment SEQIDID NO:2 NO:2 comprising comprising amino amino acids acids corresponding corresponding to atleast to at least 256amino 256 aminoacid acid residues residues of of SEQSEQ ID NO:2, ID NO:2, provided provided amino amino acids 69,acids 69,82, 78, 80, 78,102, 80, 110, 82, 102, 110, 137, 139, 137, 139, 165, 165, 189, 189,203, 203,220, 220,232 232and 248 and 248ofof SEQ SEQID IDNO:2 NO:2 are are conserved; conserved;b)b)SEQ SEQ ID ID NO:4; NO:4;

a protein a that is protein that is 98% homologous 98% homologous to SEQ to SEQ ID NO:4, ID NO:4, providedprovided amino amino acids 21, acids 21,129, 86, 127, 86, 127, 129, 154, 156, 154, 156, 182, 195,206, 182, 195, 206,218, 218,220, 220, 237, 237, 249, 249, 265,265, 255,255, 271 271 andof275 and 275 SEQof ID SEQ NO:4 ID areNO:4 are conserved; or conserved; or an an immunogenic fragment of immunogenic fragment of SEQ SEQIDIDNO:4 NO:4comprising comprising amino amino acids acids

correspondingto toatatleast corresponding least274 274amino amino acidacid residues residues of SEQ of SEQ ID NO:4, ID NO:4, providedprovided amino amino acids 21, acids 21, 86, 127, 86, 127, 129, 129, 154, 154,156, 156,182, 182,195, 195, 206, 206, 218, 218, 220, 220, 237,237, 249,249, 255, 255, 265, 265, 271275 271 and andof 275 SEQ of ID SEQ ID NO:4 are conserved; NO:4 are conserved; c) c) SEQ IDNO:6; SEQ ID NO:6;a aprotein protein that that isis98% 98% homologous to SEQ homologous to IDNO:6, SEQ ID NO:6, providedamino provided amino acids acids 14, 14, 15, 15, 32, 32, 47, 47, 58, 58, 79, 79,111, 111, 157,157, 223,223, 320,320, 350,350, 475, 475, 499, 499, 569, 624, 569, 613, 613, 624, 653, 660, 653, 660, 663, 663, 733 733 and and 734 734 of ofSEQ ID NO:6 SEQ ID are conserved; NO:6 are conserved; or or an an immunogenic fragmentofof immunogenic fragment

SEQIDID SEQ NO:6 NO:6 comprising comprising amino amino acids corresponding acids corresponding to 735 to at least at least amino735 amino acid acidofresidues residues of SEQIDID SEQ NO:6, NO:6, provided provided aminoamino acids acids 14, 15,14, 32,15, 47,32, 58,47, 58, 79,111, 79,111, 157, 157, 223, 223, 320, 320, 350, 475,350, 499,475, 499, 569, 613, 569, 613, 624, 624, 653, 653,660, 660,663, 663,733 733and and734 734ofof SEQ SEQID ID NO:6 NO:6 are conserved; d) are conserved; d)SEQ SEQ ID ID NO:8; NO:8;

a protein a that is protein that is 98% homologous 98% homologous to SEQ to SEQ ID NO:8, ID NO:8, providedprovided amino amino acids 21, acids 31, 32,21, 49,31, 64,32, 49, 64, 75, 96, 75, 96, 128, 128, 174, 174,240, 337,367, 240, 337, 367,492, 516, 492, 516, 565, 565, 586,586, 630,630,641,670,677,680,750, and751 641, 670, 677, 680, 750, and 751 of of SEQIDIDNO:8 SEQ NO:8areareconserved; conserved;ororan an immunogenic immunogenicfragment fragment of of SEQ SEQ ID ID NO:8 NO:8 comprising comprising

aminoacids amino acidscorresponding corresponding to least to at at least 752 752 amino amino acid residues acid residues of SEQ of ID SEQ NO:8,ID NO:8, provided provided aminoacids amino acids21,21, 31,31, 32,32, 49,49, 64,64, 75,75, 96,96, 128, 128, 174,174, 240,240, 337,337, 367, 367, 492, 492, 516, 516, 565, 630, 565, 586, 586, 641, 630, 641, 670, 677, 670, 677, 680, 680,750, 750,and and751751 of of SEQSEQ ID NO:8 ID NO:8 are conserved; are conserved; e) NO:10; e) SEQ ID SEQ ID NO:10;that a protein a protein that is 98% is homologoustoto SEQ 98% homologous SEQIDIDNO:10; NO:10; or or an an immunogenic immunogenic fragment fragment of SEQ of SEQ ID NO:10 ID NO:10

comprisingamino comprising amino acids acids corresponding corresponding to at to at least least 333 amino 333 amino acid residues acid residues of NO:10; of SEQ ID SEQ ID NO:10; f) SEQ f) ID NO:12; SEQ ID NO:12;aa protein protein that thatisis98% 98%homologous to SEQ homologous to ID NO:12; SEQ ID NO:12;ororananimmunogenic immunogenic

-4- fragmentofofSEQ fragment SEQ ID NO:12 ID NO:12 comprising comprising amino amino acids acids corresponding corresponding to at leastto349 at amino least 349 acid amino acid residues of residues ofSEQ SEQ ID ID NO:12; g) SEQ NO:12; g) SEQIDIDNO:14; NO:14;a protein a proteinthat that is is 98% homologoustoto SEQ 98% homologous SEQIDID

2023200517 01 Feb NO:14 NO:14 ororan an immunogenic immunogenicfragment fragmentof ofSEQ SEQ ID ID NO:14 NO:14 comprising comprising amino amino acidsacids

correspondingtotoatatleast corresponding least129 129amino amino acidacid residues residues of SEQ of SEQ ID NO:14; ID NO:14; or h) a or h) a peptide signal signal peptide linked to linked to amino aminoacids acids19-131 19-131 of SEQ of SEQ ID NO:14; ID NO:14; a protein a protein that hasthat has a signal a signal peptidepeptide linked linked to to an amino an acid sequence amino acid sequence that that isis98% 98% homologous to amino homologous to acids 19-131 amino acids 19-131 of of SEQ ID NO:14; SEQ ID NO:14;oror protein that protein that has has aa signal signal peptide peptidelinked linkedtotoananimmunogenic immunogenic fragment fragment of acids of amino amino19-131 acids of19-131 of SEQIDIDNO:14, SEQ NO:14,the thefragment fragmentcomprising comprisingatatleast least 110 110 amino acid residues amino acid residues of ofSEQ SEQ ID ID NO:14 NO:14

and linked and linkedtoto aa signal signal peptide. peptide. InInsome some embodiments embodiments the nucleic the nucleic acid molecules acid molecules are are chosen chosen fromones from onesencoding encoding proteins proteins a), a), b),b), c),c),oror d). d).

In another In aspect, the another aspect, the invention inventionincludes includesmethods methods of treating of treating an individual an individual who who has has beendiagnosed been diagnosed with with prostate prostate cancer cancer comprising comprising administering administering a nucleic a nucleic acid molecule acid molecule

describedherein described hereintotoananindividual. individual. In another In aspect, there another aspect, there are are provided providedproteins proteins selected selected from from the the group group consisting consisting of: of: a) a) SEQIDIDNO:2; SEQ NO:2;a aprotein protein that that isis98% 98% homologous to SEQ homologous to SEQIDIDNO:2, NO:2,provided providedamino amino acids69, acids 69, 78, 80, 78, 80, 82, 102, 110, 82, 102, 110, 137, 137,139, 139,165, 165,189, 189, 203, 203, 220, 220, 232 232 and and 248SEQofIDSEQ 248 of NO:2ID NO:2 are are conserved; or conserved; or an an immunogenic fragment of immunogenic fragment of SEQ SEQIDIDNO:2 NO:2 comprising comprising amino amino acids acids

correspondingtotoatatleast corresponding least261 261amino amino acidacid residues residues of SEQ of SEQ ID NO:2, ID NO:2, providedprovided amino amino acids 69, acids 69, 78, 80, 78, 80, 82, 82, 102, 102, 110, 137,139, 110, 137, 139,165, 165,189, 189, 203, 203, 220, 220, and and 232 232 248SEQofIDSEQ 248 of NO:2ID NO:2 are are conserved; b) conserved; b) SEQ ID NO:4; SEQ ID NO:4;aa protein protein that thatisis98% 98%homologous to SEQ homologous to ID NO:4, SEQ ID NO:4,provided provided aminoacids amino acids21, 21,86,86,127, 127,129, 129, 154, 154, 156,156, 182,182, 195,195, 206,206, 218, 218, 220, 220, 237, 255, 237, 249, 249,265, 255,271 265, and271 and 275 of 275 of SEQ IDNO:4 SEQ ID NO:4are areconserved; conserved; oror an an immunogenic immunogenicfragment fragmentofofSEQ SEQ ID ID NO:4 NO:4

comprisingamino comprising amino acids acids corresponding corresponding to at to at least least 274 amino 274 amino acid residues acid residues of NO:4, of SEQ ID SEQ ID NO:4, providedamino provided amino acids acids 21,21, 86,86, 127,127, 129,129, 154,154, 156, 156, 182, 182, 195, 218, 195, 206, 206, 220, 218,237, 220,249, 237,255, 249, 255, 265, 271 265, 271 and and 275 275 of of SEQ ID NO:4 SEQ ID NO:4are areconserved; conserved; c) c) SEQ SEQIDIDNO:6; NO:6;a aprotein protein that that is is98% 98%

homologous homologous to SEQ to SEQ ID NO:6, ID NO:6, provided provided amino amino acids 14, acids 14,47, 15, 32, 15,58, 32,79,111, 47, 58,157, 79,111, 223, 157, 223, 320, 350, 320, 350, 475, 475,499, 499,569, 569,613, 613,624, 624, 653, 653, 660,660, 663,663, 733 733 andof734 and 734 SEQof IDSEQ NO:6 ID areNO:6 are conserved; or conserved; or an an immunogenic fragment of immunogenic fragment of SEQ SEQIDIDNO:6 NO:6 comprising comprising amino amino acids acids

correspondingtotoatatleast corresponding least735 735amino amino acid acid residues residues of SEQ of SEQ ID NO:6, ID NO:6, providedprovided amino amino acids 14, acids 14, 15, 32, 15, 32, 47, 47, 58, 79,111, 157, 58, 79,111, 157, 223, 223,320, 320,350, 350,475, 475, 499, 499, 569, 569, 613,613, 624,624, 653, 653, 660, 660, 663, 663, 733 733 and and 734 of 734 of SEQ IDNO:6 SEQ ID NO:6are areconserved; conserved; d) d) SEQ SEQIDIDNO:8; NO:8;a a proteinthat protein that is is 98% 98% homologous homologous toto

SEQIDID SEQ NO:8, NO:8, provided provided aminoamino acids acids 21, 31,21, 32,31, 49,32, 64,49, 75,64, 96,75, 96,174, 128, 128,240, 174, 240, 337, 337, 367, 492,367, 492, 516, 565, 516, 565, 586, 586,630, 630,641, 641,670, 670,677, 677, 680, 680, 750, 750, and and 751 751 ofIDSEQ of SEQ NO:8ID NO:8 are are conserved; conserved; or an or an immunogenicfragment immunogenic fragmentofofSEQ SEQID ID NO:8 NO:8 comprising comprising amino amino acids acids corresponding corresponding to atleast to at least

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752 amino 752 aminoacid acid residues residues of of SEQSEQ ID NO:8, ID NO:8, provided provided amino amino acids 21,acids 21,49,31,64,32,75,49,96,64, 31, 32, 75, 96, 128, 174, 128, 174, 240, 240,337, 337,367, 367,492, 492,516, 516, 565, 565, 586, 586, 630, 630, 641,641, 670,670, 677, 677, 680, 680, 750,751 750, and andof751 SEQ of SEQ

2023200517 01 Feb ID NO:8 ID NO:8are are conserved; conserved; e) e) SEQ ID NO:10; SEQ ID NO:10;a aprotein protein that that isis98% 98% homologous to SEQ homologous to SEQIDID NO:10; or an NO:10; or an immunogenic immunogenicfragment fragmentofofSEQ SEQID ID NO:10 NO:10 comprising comprising amino amino acids acids

correspondingto toatatleast corresponding least333 333amino amino acidacid residues residues of SEQ of SEQ ID NO:10; ID NO:10; f) NO:12; f) SEQ ID SEQ ID a NO:12; a protein that protein thatisis 98% 98%homologous homologous to to SEQ ID NO:12; SEQ ID NO:12;oror an an immunogenic immunogenicfragment fragmentofofSEQ SEQ ID ID NO:12 comprising NO:12 comprising amino amino acids acids corresponding corresponding to at349 to at least least 349acid amino amino acid residues residues of SEQ ID of SEQ ID

NO:12; g) SEQ NO:12; g) SEQIDIDNO:14; NO:14; a proteinthat a protein that is is 98% homologoustoto SEQ 98% homologous SEQIDIDNO:14; NO:14; or or an an

immunogenicfragment immunogenic fragmentofofSEQ SEQID ID NO:14 NO:14 comprising comprising amino amino acids acids corresponding corresponding to at to at least least

129 amino 129 aminoacid acidresidues residues of of SEQSEQ ID NO:14; ID NO:14; or h) aorsignal h) a signal peptidepeptide linked linked to aminotoacids amino19-acids 19 131 of 131 ofSEQ SEQID ID NO:14; NO:14; a protein a protein that that has ahas a signal signal peptide peptide linkedlinked to an to an amino amino acid sequence acid sequence

that is that is 98% homologous 98% homologous to amino to amino acidsacids 19-13119-131 of SEQ of ID SEQ IDorNO:14; NO:14; protein or protein that has a that has signal a signal peptide linked peptide linked totoananimmunogenic immunogenic fragment of amino fragment of amino acids acids 19-131 19-131 of of SEQ ID NO:14, SEQ ID NO:14, the the fragmentcomprising fragment comprising at least at least 110110 amino amino acid acid residues residues ofIDSEQ of SEQ IDand NO:14 NO:14 linkedand to linked a to a signal peptide. signal peptide. InInsome some embodiments, embodiments, the protein the protein is selected is selected fromgroup from the the comprising: group comprising: proteins a), b), c), or d). proteins a), b), c), or d).

Someaspects Some aspectsof of thethe invention invention include include methods methods of treating of treating an individual an individual who who has beenhas been diagnosedwith diagnosed withprostate prostatecancer cancer comprising comprising delivering delivering to individual to said said individual a protein a protein described described

herein. herein.

Otheraspects Other aspectsofofthe theinvention inventionarearepharmaceutical pharmaceutical compositions compositions comprising comprising the the nucleic acid nucleic acidmolecules moleculesprovided provided herein herein and and a pharmaceutically a pharmaceutically acceptable acceptable excipient. excipient.

BRIEF DESCRIPTION BRIEF OF THE DESCRIPTION OF THE FIGURES FIGURES Figure11 shows Figure shows resultsfrom results from in in vitro vitro translationperformed translation performed to confirm to confirm the expression the expression

of the of the PSA PSA and and PSMA antigens. PSMA antigens.

Figure 2A Figure showscellular 2A shows cellular immunogenicity data. Cellular immunogenicity data. Cellular immunogenicity of PSA immunogenicity of PSA

antigens was antigens was determined determined by by Interferon-gamma ELISpot. Interferon-gamma ELISpot.

Figure 2B Figure shows cellular 2B shows cellular immunogenicity data. Cellular immunogenicity data. Cellular immunogenicity of PSA immunogenicity of PSA

Figures 3A-C Figures showsCD4+ 3A-C shows CD4+ T cellresponses T cell responsesasascharacterized characterized by by flow flow cytometry cytometry by by

displayinggraphs displaying graphsshowing showing PSA-specific PSA-specific (left (left panel), panel), PSMA-specific PSMA-specific (middle (middle panel) panel) and total and total vaccine-specific(right vaccine-specific (rightpanel) panel)cytokine cytokineproduction: production: % IFNy % IFNy producing producing CD4+ T CD4+ T cells cells (Fig. (Fig. 3A); %%IL-2 3A); IL-2 producing producing CD4+ CD4+T Tcells cells(Fig. (Fig. 3B); 3B); and and % % TNFc producing TNF producing CD4+ CD4+ T cells T cells (Fig. (Fig.

3C). 3C).

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Feb 2023 Figures 4A-C Figures CD8+ showsCD8+ 4A-C shows T cellresponses T cell characterized by responsesasascharacterized by flow flow cytometry cytometry by by

displayinggraphs displaying graphsshowing showing PSA-specific PSA-specific (left (left panel), panel), PSMA-specific PSMA-specific (middle (middle panel) panel) and total and total vaccine-specific(right vaccine-specific (rightpanel) panel)cytokine cytokineproduction: production: %IFNy % IFNy producing producing CD8+(Fig. CD8+ T cells T cells (Fig. 4A); %%IL-2 4A); IL-2 producing producing CD8+ CD8+T T cells(Fig. cells (Fig. 4B); 4B); and and % TNFaproducing % TNFa producingCD8+ CD8+ T cells(Fig. T cells (Fig. 2023200517 01 4C). 4C).

Figures5A-B Figures 5A-B shows shows ELISA ELISA data data for for PSA-specific PSA-specific antibodies one week one antibodies week after after the final the final immunization.(Fig immunization. (Fig 5A)5A) PSA PSA IgG endpoint IgG endpoint titers.titers. (FigRepresentative (Fig 5B) 5B) Representative IgG titration IgG titration

curves. curves.

DETAILED DESCRIPTION DETAILED DESCRIPTION Providedherein Provided hereinareareconsensus consensus sequence sequence prostate prostate proteins proteins and isolated and isolated nucleic nucleic acid acid

moleculesthat molecules thatencode encode them, them, and and in particular, in particular, the the prostate prostate antigens antigens prostate prostate specific specific antigen antigen

(PSA), prostate (PSA), prostatespecific specificmembrane membrane antigen antigen (PSMA), (PSMA), six-transmembrane six-transmembrane epithelial epithelial antigen of antigen of the prostate the antigen (STEAP) prostate antigen (STEAP)and and prostate prostate specific specific stem stem cell antigen cell antigen (PSCA). (PSCA).

Theprostate The prostatecancer cancerantigens antigens described described herein herein are are consensus consensus sequences sequences derivedderived from a from a pool of pool ofhomologous homologous antigens antigens from from across across multiple multiple species, species, including including the specie the specie that thethat the vaccineisis targeted vaccine targeted for. for. The Theselected selectedspecies species from from which which antigen antigen sequences sequences are aligned are aligned to to formaaconsensus form consensus shallbe be shall chosen chosen based based on close on close proximity proximity of the of the species species on a phylogenic on a phylogenic

tree, e.g., tree, e.g.,H.sapiens (humans),M.mulatta H.sapiens (humans), M.mulatta (rhesus (rhesus macaques), macaques), and M.fascicularis and M.fascicularis (cynomolgus (cynomolgus

monkey).TheThe monkey). consensus consensus antigen antigen is notisidentical not identical to thetonative the native prostate prostate antigen antigen buthave but will will have close identity, close identity, which sequences which sequences share share at least at least 85%, 85%, and and preferably preferably 90%, 90%, 91%,93%, 91%, 92%, 92%, 93%, 94%, 95%, 94%, 95%, 96%, 96%,97%, 97%, 98%, 98%, or or 99%99% identity.These identity. These described described consensus consensus cancer cancer antigensare antigens are able to able to break toleranceininthe break tolerance thetargeted targetedspecie specie(or (orcause causeautoimmunity) autoimmunity) and generate and generate an an effective immune effective immune response response against against the the prostate prostate cancer cancer antigen. antigen. Provided Provided herein herein are are methods methods to generate to generate aa consensus consensuscancer cancer antigen antigen based based DNA vaccine. DNA vaccine.

Aspects Aspects ofofthe thepresent presentinvention invention include include nucleic nucleic acid acid molecules molecules comprising comprising a codinga coding

a protein a protein that that is is 98% homologous 98% homologous to SEQ to SEQ ID NO:4, ID NO:4, providedprovided amino amino acids 21, acids 21,129, 86, 127, 86, 127, 129, 154, 156, 154, 156, 182, 195,206, 182, 195, 206,218, 218,220, 220, 237, 237, 249, 249, 265,265, 255,255, 271 271 andof275 and 275 SEQof IDSEQ NO:4 ID areNO:4 are

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Feb 2023 conserved; or conserved; or an an immunogenic fragment of immunogenic fragment of SEQ SEQIDIDNO:4 NO:4comprising comprising amino amino acids acids

correspondingto toatatleast corresponding least274 274amino amino acidacid residues residues of SEQ of SEQ ID NO:4, ID NO:4, providedprovided amino amino acids 21, acids 21, 86, 127, 86, 127, 129, 129, 154, 154,156, 156,182, 182,195, 195, 206, 206, 218, 218, 220, 220, 237,237, 249,249, 255, 255, 265, 265, 271275 271 and andof 275 SEQ of ID SEQ ID NO:4 are conserved; NO:4 are conserved; c) c) SEQ IDNO:6; SEQ ID NO:6;a aprotein protein that that isis98% 98% homologous to SEQ homologous to IDNO:6, SEQ ID NO:6, 2023200517 01 providedamino provided amino acids acids 14, 14, 15, 15, 32, 32, 47, 47, 58, 58, 79,111, 79,111, 157, 157, 223, 223, 320, 320, 350, 350, 475, 569, 475, 499, 499,613, 569,624, 613, 624, 653, 660, 653, 660, 663, 663, 733 733 and and 734 734 of ofSEQ ID NO:6 SEQ ID are conserved; NO:6 are conserved; or or an an immunogenic fragmentofof immunogenic fragment

SEQIDID SEQ NO:6 NO:6 comprising comprising amino amino acids corresponding acids corresponding to 735 to at least at least amino735 amino acid acidofresidues residues of SEQIDID SEQ NO:6, NO:6, provided provided aminoamino acids acids 14, 15,14, 32,15, 47,32, 58,47, 58, 79,111, 79,111, 157, 157, 223, 223, 320, 320, 350, 475,350, 499,475, 499, 569, 613, 569, 613, 624, 624, 653, 653,660, 660,663, 663,733 733and 734ofof and734 SEQ SEQID IDNO:6 NO:6 are are conserved; conserved; d) SEQ ID d)SEQ ID NO:8; NO:8;

a protein a protein that that is is 98% homologous 98% homologous to SEQ to SEQ ID NO:8, ID NO:8, providedprovided amino amino acids 21, acids 21,49,31,64,32, 31, 32, 49, 64, 75, 96, 75, 96, 128, 128, 174, 174,240, 337,367, 240, 337, 367,492, 516, 492, 516, 565, 565, 586,586, 630,630,641,670,677,680,750, and751 641, 670, 677, 680, 750, and 751 of of SEQIDIDNO:8 SEQ NO:8areareconserved; conserved;ororan an immunogenic immunogenicfragment fragment of of SEQ SEQ ID ID NO:8 NO:8 comprising comprising

aminoacids amino acidscorresponding corresponding to least to at at least 752 752 amino amino acid residues acid residues of SEQ of ID SEQ NO:8,ID NO:8, provided provided aminoacids amino acids21,21, 31,31, 32,32, 49,49, 64,64, 75,75, 96,96, 128, 128, 174,174, 240,240, 337,337, 367, 367, 492, 492, 516, 516, 565, 630, 565, 586, 586, 641, 630, 641, 670, 677, 670, 677, 680, 680,750, 750,and and751751 of of SEQSEQ ID NO:8 ID NO:8 are conserved; are conserved; e) NO:10; e) SEQ ID SEQ ID NO:10;that a protein a protein that is 98% is homologoustoto SEQ 98% homologous SEQIDIDNO:10; NO:10; or or anan immunogenic immunogenic fragment fragment of SEQ of SEQ ID NO:10 ID NO:10

comprisingamino comprising amino acids acids corresponding corresponding to at to at least least 333 amino 333 amino acid residues acid residues of NO:10; of SEQ ID SEQ ID NO:10; f) SEQ f) ID NO:12; SEQ ID NO:12;aa protein protein that thatisis98% 98%homologous to SEQ homologous to ID NO:12; SEQ ID NO:12;ororananimmunogenic immunogenic fragmentofofSEQ fragment SEQ ID NO:12 ID NO:12 comprising comprising amino amino acids acids corresponding corresponding to at leastto349 at amino least 349 acid amino acid residues of residues ofSEQ SEQ ID ID NO:12; g) SEQ NO:12; g) SEQIDIDNO:14; NO:14;a protein a proteinthat that is is 98% homologoustoto SEQ 98% homologous SEQIDID NO:14; or an NO:14; or an immunogenic immunogenicfragment fragmentofofSEQ SEQID ID NO:14 NO:14 comprising comprising amino amino acids acids

correspondingto toatatleast corresponding least129 129amino amino acidacid residues residues of SEQ of SEQ ID NO:14; ID NO:14; or an immunogenic or an immunogenic

fragment of fragment of SEQ IDNO:14 SEQ ID NO:14comprising comprisingatatleast least 129 129 amino amino acid acid residues residues of of SEQ ID NO:14; SEQ ID NO:14; or h) or a signal h) a signal peptide linked toto amino peptide linked aminoacids acids19-131 19-131 of SEQ of SEQ ID NO:14; ID NO:14; a protein a protein that hasthat a has a signal peptide signal peptide linked linkedtotoananamino amino acid acid sequence sequence that that is 98% is 98% homologous homologous to aminotoacids amino 19- acids 19 131 of 131 ofSEQ SEQID ID NO:14; NO:14; or protein or protein that that has ahas a signal signal peptide peptide linkedlinked to an to an immunogenic immunogenic

fragmentofofamino fragment amino acids acids 19-131 19-131 of ID of SEQ SEQ ID NO:14, NO:14, the fragment the fragment comprisingcomprising at at least 110 least 110 aminoacid amino acidresidues residuesofof SEQ SEQ ID NO:14 ID NO:14 and linked and linked to a signal to a signal peptide. peptide. Two consensus Two consensus protein protein sequences for sequences for PSA are disclosed: PSA are disclosed: PSA PSA Consensus Antigen sequence Consensus Antigen sequence 11 (SEQ (SEQIDIDNO:2) NO:2)andand PSAConsensus PSA ConsensusAntigen Antigensequence sequence2 (SEQ 2 (SEQ ID ID NO:4). NO:4). Two Two consensus consensus protein protein sequences sequences for for PSMA PSMA aredisclosed: are disclosed: PSMA PSMA Consensus Consensus Antigen Antigen sequence sequence 1 (SEQ 1 (SEQ ID NO:6) ID NO:6) and and PSMA PSMA Consensus Antigen Consensus Antigensequence sequence22(SEQ (SEQIDIDNO:8). NO:8).TwoTwo consensus consensus protein protein sequences sequences forfor

STEAP(also STEAP (alsoreferred referred to to herein hereinasasSTEAPI) are disclosed: STEAP1) are disclosed:STEAP ConsensusAntigen STEAP Consensus Antigen sequence 11 (SEQ sequence (SEQ ID IDNO:10) NO:10)and andSTEAP STEAP Consensus Consensus Antigen Antigen sequence sequence 2 (SEQ 2 (SEQ ID NO:12). ID NO:12).

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Feb 2023 One consensus One protein sequence consensus protein for PSCA sequence for is disclosed: PSCA is disclosed: PSCA ConsensusAntigen PSCA Consensus Antigensequence sequence (SEQ IDIDNO:14). (SEQ NO:14).SEQSEQ ID ID NO:14 NO:14 includes includes an IgE an IgE signal signal some peptide.InInsome peptide. embodiments, embodiments, a a PSCAConsensus PSCA Consensus antigenmay antigen may includeamino include amino acids19-131 acids 19-131of of SEQ SEQ ID ID NO:14 NO:14 linked linked to to a a signal sequence signal sequence other otherthan thanthe IgE the signal IgE in SEQ signal IDID in SEQ NO:14. NO:14.InInsome someembodiments the embodiments the

2023200517 01 nucleic acid nucleic acid molecules moleculesareare chosen chosen fromfrom ones ones encoding encoding proteins proteins a), b),a), c),b),orc),d),orabove. d), above. In In other embodiments other embodiments the the nucleic nucleic acidacid molecules molecules are encoding are ones ones encoding one proteins one or more or more proteins selected from selected fromthe thegroup groupcomprising: comprising: at least at least oneone selected selected fromfrom ones ones encoding encoding either either proteins proteins

a) or a) or b), b), and and at at least leastone one selected fromones selected from onesencoding encoding either either proteins proteins c) d). c) or or d). Thenucleic The nucleicacid acidmolecules moleculescan can further further be molecules be molecules encoding encoding one or one more or more proteins proteins

selected from selected from the thegroup groupcomprising: comprising:SEQ SEQ ID ID NO:2; SEQIDIDNO:4; NO:2; SEQ NO:4;SEQSEQ ID ID NO:6; NO:6; SEQ SEQ ID ID NO:8; SEQIDIDNO:10; NO:8; SEQ NO:10;SEQSEQ ID NO:12; ID NO:12; or SEQ or SEQ ID NO:14; ID NO:14; and preferably, and preferably, SEQ SEQ ID ID NO:2; NO:2;

SEQIDIDNO:4; SEQ NO:4;SEQ SEQ ID ID NO:6; NO:6; or SEQ or SEQ ID NO:8. ID NO:8. In some In some embodiments, embodiments, the nucleic the nucleic acid acid moleculeofofcan molecule canbebeones ones that that encode encode one one or more or more proteins proteins selected selected from from the the group group comprising:atatleast comprising: leastone oneselected selectedfrom from either either SEQSEQ ID NO:2 ID NO:2 or SEQ or ID SEQ NO:4,ID andNO:4, andone at least at least one selected from selected from either eitherSEQ SEQ ID ID NO:6 or SEQ NO:6 or IDNO:8. SEQ ID NO:8. In another In another aspect, aspect, there there are are provided providedproteins proteins selected selected from from the the group group consisting consisting of: of: a) a) SEQIDIDNO:2; SEQ NO:2;a aprotein protein that that isis98% 98% homologous to SEQ homologous to SEQIDIDNO:2, NO:2,provided providedamino amino acids69, acids 69, 80, 82, 78, 80, 78, 82, 102, 102, 110, 110, 137, 137,139, 139,165, 165,189, 189, 203, 203, 220, 220, 232 232 and and 248SEQofIDSEQ 248 of areNO:2 NO:2ID are conserved; or conserved; or an an immunogenic fragment of immunogenic fragment of SEQ SEQIDIDNO:2 NO:2 comprising comprising amino amino acids acids

correspondingto toatatleast corresponding least256 256amino amino acidacid residues residues of SEQ of SEQ ID NO:2, ID NO:2, providedprovided amino amino acids 69, acids 69, 78, 80, 78, 80, 82, 102, 110, 82, 102, 110, 137, 137,139, 139,165, 165,189, 189, 203, 203, 220, 220, 232 232 and and 248SEQofIDSEQ 248 of NO:2ID NO:2 are are conserved; b) conserved; b) SEQ ID NO:4; SEQ ID NO:4;aa protein protein that thatisis98% 98%homologous to SEQ homologous to ID NO:4, SEQ ID NO:4,provided provided aminoacids amino acids21, 21,86,86,127, 127,129, 129, 154, 154, 156,156, 182,182, 195,195, 206,206, 218, 218, 220, 220, 237, 255, 237, 249, 249,265, 255,271 265, and271 and 275 of 275 of SEQ IDNO:4 SEQ ID NO:4are areconserved; conserved; oror an an immunogenic immunogenicfragment fragmentofofSEQ SEQ ID ID NO:4 NO:4

homologous homologous to SEQ to SEQ ID NO:6, ID NO:6, provided provided amino amino acids 14, acids 14,47, 15, 32, 15,58, 32,79,111, 47, 58,157, 79,111, 223, 157, 223, 320, 350, 320, 350, 475, 475,499, 499,569, 569,613, 613,624, 624, 653, 653, 660, 660, 663,663, 733 733 andof734 and 734 SEQof IDSEQ NO:6 ID areNO:6 are conserved; or conserved; or an an immunogenic fragment of immunogenic fragment of SEQ SEQIDIDNO:6 NO:6 comprising comprising amino amino acids acids

correspondingtotoatatleast corresponding least735 735amino amino acidacid residues residues of SEQ of SEQ ID NO:6, ID NO:6, providedprovided amino amino acids 14, acids 14, 15, 32, 15, 32, 47, 47, 58, 157, 223, 79,111, 157, 58, 79,111, 223,320, 320,350, 350,475, 475, 499, 499, 613,613, 569, 569, 624,624, 653, 653, 660, 660, 663, 663, 733 733 and and 734 of 734 of SEQ IDNO:6 SEQ ID NO:6are areconserved; conserved; d) d) SEQ SEQIDIDNO:8; NO:8;a a proteinthat protein that is is 98% 98% homologous homologous toto

SEQIDID SEQ NO:8, NO:8, provided provided aminoamino acids acids 21, 31,21, 32,31, 49,32, 64,49, 75,64, 96,75, 96,174, 128, 128,240, 174, 240, 337, 337, 367, 492,367, 492,

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Feb 2023 516, 565, 516, 565, 586, 586,630, 641,670, 630,641, 670,677, 677, 680, 680, 750, 750, and and 751SEQ 751 of NO:8ID ofIDSEQ NO:8 are are conserved; conserved; or an or an immunogenicfragment immunogenic fragmentofofSEQ SEQID ID NO:8 NO:8 comprising comprising amino amino acids acids corresponding corresponding to atleast to at least 752 amino 752 aminoacid acid residues residues of of SEQSEQ ID NO:8, ID NO:8, provided provided amino amino acids 21,acids 21,49,31,64,32,75,49,96,64, 31, 32, 75, 96, 128, 174, 128, 174, 240, 240,337, 337,367, 367,492, 492,516, 516, 565, 565, 586, 586, 630, 630, 641,641, 670,670, 677, 677, 680, 680, 750,751 750, and andof751 SEQ of SEQ 2023200517 01 ID NO:8 ID NO:8are are conserved; conserved; e) e) SEQ ID NO:10; SEQ ID NO:10;a aprotein protein that that isis98% 98% homologous to SEQ homologous to SEQIDID NO:10; or an NO:10; or an immunogenic immunogenicfragment fragmentofofSEQ SEQID ID NO:10 NO:10 comprising comprising amino amino acids acids

that is that is 98% homologous 98% homologous to amino to amino acids acids 19-13119-131 of SEQ of ID SEQ IDorNO:14; NO:14; protein or protein that has a that has signal a signal peptide linked peptide linked totoananimmunogenic immunogenic fragment of amino fragment of amino acids acids 19-131 19-131 of of SEQ ID NO:14, SEQ ID NO:14, the the fragmentcomprising fragment comprising at least at least 110110 amino amino acid acid residues residues ofIDSEQ of SEQ IDand NO:14 NO:14 linkedand to linked a to a signal peptide. signal peptide. InInsome some embodiments, embodiments, the protein the protein is selected is selected from from the the comprising: group group comprising: proteins a), proteins a), b), b), c), c),or ord). d). In In other other embodiments embodiments thethe proteins proteins areare ones ones encoding encoding onemore one or or more proteins selected proteins selected from fromthe thegroup group comprising: comprising: at least at least one one selected selected from from either either proteins proteins a) or a) or b), and b), at least and at least one selected from one selected fromeither eitherproteins proteinsc)c)orord). d). Theproteins The proteinscan canfurther furtherbebeproteins proteinsselected selected from from the the group group comprising: comprising: SEQ IDSEQ ID NO:2; SEQIDIDNO:4; NO:2; SEQ NO:4; SEQSEQ ID NO:6; ID NO:6; SEQ SEQ ID NO:8; ID NO:8; SEQ IDSEQ ID NO:10; NO:10; SEQ IDorNO:12; or SEQ ID NO:12;

SEQIDIDNO:14; SEQ NO:14;andandpreferably, preferably, SEQ SEQIDIDNO:2; NO:2; SEQSEQ ID NO:4; ID NO:4; SEQ SEQ ID NO:6; ID NO:6; or SEQorID SEQ ID NO:8. NO:8. In In some some embodiments, embodiments, the proteins the proteins can be can onesbe ones selected selected from the from groupthe group comprising: comprising:

at least at least one one selected fromeither selected from eitherSEQ SEQID ID NO:2 NO:2 or ID or SEQ SEQ IDand NO:4, NO:4, and at at least oneleast one selected selected from either from either SEQ ID NO:6 SEQ ID NO:6oror SEQ SEQIDIDNO:8. NO:8. Nucleic acidcoding Nucleic acid coding sequences sequences havehave been been generated generated to improve to improve and optimize and optimize

expression. The expression. The codons codons usedused in these in these nucleic nucleic acid acid molecules molecules were selected were selected to generate to generate RNA RNA havingreduced having reducedsecondary secondary structure structure formation formation due due to to intramolecular intramolecular hybridization. hybridization. Nucleic Nucleic acid sequences acid sequences encoding encoding PSA ConsensusAntigen PSA Consensus Antigensequence sequence1 (SEQ 1 (SEQ ID ID NO:1) NO:1) and and PSAPSA

Consensus Antigen Consensus Antigensequence sequence22(SEQ (SEQIDIDNO:3) NO:3) areare disclosed. Likewise, disclosed. Likewise,nucleic nucleic acid acid coding coding

sequence for sequence for PSMA Consensus PSMA Consensus Antigen Antigen sequence sequence 1 (SEQ 1 (SEQ ID NO:5 ID NO:5 of nucleotides of nucleotides 1-2250 1-2250 of of SEQIDIDNO:5) SEQ NO:5)and andPSMA PSMA Consensus Consensus Antigen Antigen sequence sequence 2 (SEQ 2 (SEQ ID or ID NO:7 NO:7 or nucleotides nucleotides 1- 1

2301 of 2301 of SEQ SEQIDIDNO:7) NO:7)asaswell wellasas STEAP STEAP Consensus Consensus Antigen Antigen sequence sequence 1 (SEQ 1 (SEQ ID NO:9), ID NO:9),

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Feb 2023 STEAPConsensus STEAP Consensus Antigen Antigen sequence sequence 2 (SEQ 2 (SEQ ID NO:11) ID NO:11) and PSCA and PSCA Consensus AntigenAntigen Consensus sequence(SEQ sequence (SEQ ID NO:13) ID NO:13) are provided. are provided. Also provides Also provides are acid are nucleic nucleic acid sequences sequences that are that are 98%homologous 98% homologousto to SEQ SEQ ID ID NO:1 NO:1 and and encode encode either either PSAPSA Consensus Consensus Antigen Antigen sequence sequence 1 1 (SEQ IDIDNO:2) (SEQ NO:2)orora aprotein protein up up to to 98% homologoustotoSEQ 98% homologous SEQIDID NO:2, NO:2, preferablyincluding preferably including

2023200517 01 aminoacids amino acids69, 69,78,78,80,80,82,82,102, 102,110, 110, 137, 137, 139,139, 165,165, 189,189, 203,203, 220, 220, 232248 232 and andof248 SEQ of ID SEQ ID NO:2, and nucleic NO:2, and nucleic acid acid sequences sequences that thatare 98% are 98%homologous to SEQ homologous to ID NO:3 SEQ ID NO:3and andencode encode either PSA either PSA Consensus Antigensequence Consensus Antigen sequence22 (SEQ (SEQIDIDNO:4) NO:4)or or a aprotein protein up up to to 98% 98% homologous homologous to SEQ to SEQ ID NO:4, ID NO:4, preferably preferably including including amino amino acids 21, acids 21, 129, 86, 127, 86, 127, 129, 154, 156, 154, 156, 182, 195, 182, 195, 206, 206,218, 218,220, 220,237, 237,249, 249, 255, 255, 265, 265, 271 271 and and 275SEQofIDSEQ 275 of IDLikewise, NO:4. NO:4. Likewise, nucleic acid nucleic acidsequences sequencesthat thatare 98% are 98%homologous homologous to to nucleotides nucleotides2250 2250ofofSEQ SEQ ID ID NO:5 and NO:5 and

encode either encode either PSMA ConsensusAntigen PSMA Consensus Antigen sequence sequence 1 (SEQ 1 (SEQ ID ID NO:6) NO:6) or aorprotein a protein upup toto98% 98% homologous homologous to SEQ to SEQ ID NO:6, ID NO:6, preferably preferably including including amino amino acids 14, acids 14,47,15,58,32,79,111, 15, 32, 47, 58, 79,111, 157, 223, 157, 223, 320, 320, 350, 350,475, 475,499, 569, 499,569, 613, 613, 624, 624, 653,653, 660,660, 663,663, andof734 733734 733 and SEQ of ID SEQ NO:6,ID or NO:6, or nucleic acid nucleic acidsequences sequencesthat thatare 98% are 98%homologous homologous to to nucleotides nucleotides2301 2301ofofSEQ SEQ ID ID NO:7 and NO:7 and

encode either encode either PSMA ConsensusAntigen PSMA Consensus Antigen sequence sequence 1 (SEQ 1 (SEQ ID ID NO:8) NO:8) or aorprotein a protein upup toto98% 98% homologous homologous to SEQ to SEQ ID NO:8, ID NO:8, preferably preferably including including amino amino acids 21, acids 21,49,31,64,32,75,49, 31, 32, 96,64, 75, 96, 128, 174, 128, 174, 240, 240,337, 337,367, 367,492, 492,516, 516,565, 565, 586, 586, 630, 630, 641,641, 670,670, 677, 677, 680, 680, 750,751 750, and andof751 SEQ of SEQ ID NO:8 ID NO:8asas well well as as nucleotides nucleotides 98% 98% homologous to SEQ homologous to SEQIDIDNONO 9 and 9 and encode encode eitherSTEAP either STEAP Consensus Antigen Consensus Antigensequence sequence1 1(SEQ (SEQIDIDNO:10) NO:10) or or a proteinthat a protein that is is up up toto98% 98% homologous to homologous to

SEQIDIDNO:10, SEQ NO:10,nucleotides nucleotides98% 98%homologous homologous to SEQ to SEQ ID NO:11 ID NO:11 and encode and encode either either STEAP STEAP

Consensus Antigen Consensus Antigensequence sequence22(SEQ (SEQIDIDNO:12) NO:12) or or a proteinthat a protein that is is up up toto98% 98% homologous to homologous to

SEQIDIDNO:12, SEQ NO:12,and andnucleotides nucleotides98% 98% homologous homologous to SEQ to SEQ ID NO:13 ID NO:13 and encodes and encodes with with PSCA PSCA Consensus Antigen Consensus Antigensequence sequence(SEQ (SEQIDIDNO:14) NO:14) or or a proteinthat a protein that is is up up to to98% 98% homologous to homologous to

SEQIDIDNO:14. SEQ NO:14.In In some some embodiments embodiments nucleic nucleic acid acid molecules molecules encode encode a proteinthat a protein that comprises an comprises an IgE IgE signal signalpeptide peptide(for example, (for SEQ example, SEQ ID IDNO:3 NO:3 which encodes SEQ which encodes SEQIDIDNO:4; NO:4; nucleotides 1-2301 nucleotides 1-2301 of of SEQ ID NO:7 SEQ ID NO:7which whichencodes encodesSEQ SEQ ID ID NO:8; NO:8; SEQ SEQ ID NO: ID NO:11 1 which which

encodes SEQ encodes SEQIDIDNO:12, NO:12,andand SEQ SEQ ID NO:13 ID NO:13 which which encodes encodes SEQ SEQ ID ID NO:14). NO:14).

Compositions comprising Compositions comprising nucleic nucleic acid acid molecules molecules which comprise the which comprise the coding coding

sequencesofofthe sequences theisolated isolatednucleic nucleicacid acid molecules molecules provided provided herein herein may bemay be useful useful for inducing for inducing

immune immune responses responses against against a prostate a prostate protein protein when when administered administered into an into an animal. animal.

Compositions Compositions containing containing one one or more or more of these of these nucleic nucleic acid sequences acid sequences mayasbe may be used used as vaccinesororvaccine vaccines vaccinecomponents components to prophylactically to prophylactically or therapeutically or therapeutically immunize immunize against against prostate cancer. prostate cancer. Likewise, Likewise, compositions compositions comprising comprising consensus consensus proteinsproteins may be may be useful foruseful for inducingimmune inducing immune responses responses against against a prostate a prostate protein protein when administered when administered into an into an animal. animal.

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Feb 2023 Combinations of Combinations of compositions compositions comprising comprising nucleic nucleic acid acid molecules molecules which comprise the which comprise the coding coding

sequencesofofthe sequences theisolated isolatednucleic nucleicacid acid molecules molecules provided provided herein herein may bemay be useful useful to to induce induce immune immune responses responses against against a prostate a prostate protein protein andcollectively and may may collectively be usedbeasused as vaccines vaccines or or vaccinecomponents vaccine components to prophylactically to prophylactically or therapeutically or therapeutically immunize immunize against against prostateprostate cancer. cancer. 2023200517 01 Likewise, compositions Likewise, comprising consensus compositions comprising consensus proteins proteins may be useful may be useful for forinducing inducingimmune immune

responsesagainst responses againsta aprostate prostateprotein proteinwhen when administered administered into into an animal. an animal. Compositions Compositions

containingone containing oneorormore more of of these these consensus consensus proteins proteins may may be beasused used as vaccines vaccines or or vaccine vaccine components components to to prophylactically prophylactically or therapeutically or therapeutically immunize immunize againstagainst prostate prostate cancer. cancer.

Vaccines areprovided Vaccines are provided which which comprises comprises nucleic nucleic acid sequences acid sequences providedprovided herein. In herein. In

some embodiments, some embodiments,vaccines vaccinesare are provided provided which whichcomprises comprisesnucleic nucleic acid acid sequences sequences encoding encoding one or one or more moreconsensus consensus prostate prostate antigens antigens selected selected from from the group the group consisting consisting of: consensus of: consensus

PSAantigen PSA antigen 1, 1, consensus consensus PSA antigen 2, PSA antigen 2, consensus consensus PSMA antigen1,1, consensus PSMA antigen consensusPSMA PSMA antigen 2,2,consensus antigen consensus STEAP antigen 1, STEAP antigen 1, consensus STEAPantigen consensus STEAP antigen2,2, and and consensus consensus PSCA. PSCA. Methodsof Methods of inducing inducing immune immuneresponses responsesusing usingnucleic nucleic acid acid sequences sequences encoding one or encoding one or more more

prostate antigens prostate antigensselected selectedfrom fromthethegroup group consisting consisting of: of: consensus consensus PSA antigen PSA antigen 1, consensus 1, consensus

PSAantigen PSA antigen 2, 2, consensus consensus PSMA antigen1,1, consensus PSMA antigen consensusPSMA PSMA antigen antigen 2, 2, consensusSTEAP consensus STEAP antigen 1,1,consensus antigen consensus STEAP antigen 2, STEAP antigen 2, and and consensus consensus PSCA. PSCA.

Vaccines which comprise Vaccines which compriseone oneoror more moreof of consensus consensus PSA PSAantigen antigen1,1, consensus consensus PSA PSA antigen 2,2,consensus antigen consensus PSMA antigen 1, PSMA antigen 1, consensus consensus PSMA antigen2,2,consensus PSMA antigen consensusSTEAP STEAP antigen 1,1,consensus antigen consensus STEAP antigen 2, STEAP antigen 2, and and consensus consensus PSCA are provided. PSCA are provided. Methods Methodsofof inducing immune inducing responsesusing immune responses using one one or or more more of of consensus consensus PSA PSAantigen antigen 1, 1, consensus PSA consensus PSA

antigen 2,2,consensus antigen consensus PSMA antigen 1,1, consensus PSMA antigen consensus PSMA antigen2,2,consensus PSMA antigen consensusSTEAP STEAP antigen 1,1, consensus antigen consensusSTEAP STEAP antigen antigen 2, and2,consensus and consensus PSCA PSCA are are also provided. also provided.

Methodsof of Methods protecting protecting an an individual individual against against prostate prostate cancer cancer or ofortreating of treating an an individual who individual whohashas been been identified identified as as having having prostate prostate cancer cancer are provided. are provided. The methods The methods

comprisethe comprise thestep stepof: of:administering administeringto to said said individual individual an an effective effective amount amount oforone of one or more more nucleic acid nucleic acid molecules moleculescomprising comprising one one or more or more nucleic nucleic acid sequences acid sequences providedprovided herein. herein. In In somemethods, some methods,thethe delivery delivery of the of the nucleic nucleic acidacid molecules molecules is facilitated is facilitated by electroporation by electroporation of of the targeted the tissue or targeted tissue or the tissue that the tissue that receives receives the nucleic acid the nucleic acid molecules. molecules.TheThe nucleic nucleic acidacid

sequenceisisexpressed sequence expressedin in cellsofofthe cells theindividual individualandand an an immune immune response response is induced is induced against against

the prostate the protein encoded prostate protein encodedbyby thethe nucleic nucleic acid acid sequence. sequence.

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Feb 2023 1. Definitions. 1. Definitions. Theterminology The terminology used used herein herein is for is for thethe purpose purpose of describing of describing particular particular embodiments embodiments

only and only andisis not not intended intendedtotobebelimiting. limiting.AsAs used used in the in the specification specification and and the the appended appended claims, claims,

the singular the singular forms forms"a," "a,""an" "an"andand "the" "the" include include plural plural referents referents unless unless the the context context clearly clearly

2023200517 01 dictates otherwise. dictates otherwise.

For recitation For recitation of of numeric numericranges ranges herein, herein, each each intervening intervening number number there there between between with with the same the samedegree degreeof of precision precision is is explicitlycontemplated. explicitly contemplated. For For example, example, forrange for the the range ofthe of 6-9, 6-9, the numbers7 and numbers 7 and 8 are 8 are contemplated contemplated in addition in addition to 6 to and6 9, and and9,for andthe forrange the range 6.0-7.0, 6.0-7.0, the the numbers6.0, numbers 6.0,6.1, 6.1,6.2, 6.2,6.3, 6.3, 6.4, 6.5, 6.6, 6.4, 6.5, 6.6, 6.7, 6.7, 6.8, 6.8, 6,9, 6,9,and and 7.0 7.0 are are explicitly explicitly contemplated. contemplated.

a. Adjuvant a. Adjuvant "Adjuvant" as "Adjuvant" as used used herein herein means any molecule means any added to molecule added to the the DNA plasmidvaccines DNA plasmid vaccines describedherein described hereintotoenhance enhancethethe immunogenicity immunogenicity of theof the antigens antigens encodedencoded by by the DNA the DNA plasmidsand plasmids andthetheencoding encoding nucleic nucleic acidacid sequences sequences described described hereinafter. hereinafter.

b. Antibody b. Antibody "Antibody"as as "Antibody" used used herein herein means means an antibody an antibody of classes of classes IgG,IgA, IgG, IgM, IgM,IgDIgA, IgD or IgE, or IgE, or fragments, or fragments fragments, fragments or or derivatives derivatives thereof, thereof, including including Fab,Fab, F(ab')2, F(ab')2, Fd, single Fd, and and single chainchain

antibodies, diabodies, antibodies, diabodies,bispecific bispecificantibodies, antibodies,bifunctional bifunctionalantibodies antibodies and and derivatives derivatives thereof. thereof.

Theantibody The antibodycancan be be an an antibody antibody isolated isolated from from the serum the serum samplesample of mammal, of mammal, a polyclonal a polyclonal

antibody, affinity antibody, affinity purified purified antibody, antibody,orormixtures mixtures thereof thereof which which exhibits exhibits sufficient sufficient binding binding

specificity to specificity to a desired epitope or desired epitope or aa sequence sequencederived derived therefrom. therefrom.

c. Coding c. Coding Sequence Sequence "Codingsequence" "Coding sequence" or "encoding or "encoding nucleic nucleic acid" acid" asherein as used used herein means means the the acids nucleic nucleic acids (RNAororDNA (RNA DNA molecule) molecule) thatcomprise that comprise a a nucleotide sequence nucleotide sequencewhich whichencodes encodesa aprotein. protein. The The codingsequence coding sequencecancan further further include include initiation initiation and and termination termination signals signals operably operably linkedlinked to to regulatory elements regulatory elementsincluding including a promoter a promoter and polyadenylation and polyadenylation signal signal capablecapable of directing of directing

expressionininthe expression thecells cells of ofan an individual individualorormammal mammal to whom to whom the nucleic the nucleic acid isacid is administered. administered.

d. Complement d. Complement "Complement"oror"complementary" "Complement" "complementary"as as used used hereinmeans herein means a nucleicacid a nucleic acidcan canmean mean Watson-Crick Watson-Crick (e.g.,A-T/U (e.g., A-T/U and and C-G) C-G) or Hoogsteen or Hoogsteen base between base pairing pairing nucleotides between nucleotides or or nucleotide analogs nucleotide analogsofofnucleic nucleicacid acid molecules. molecules.

e. Consensus e. ConsensusororConsensus Consensus Sequence Sequence

"Consensus" or "Consensus" or "consensus "consensus sequence" sequence" as as used used herein herein means means aa polypeptide polypeptide sequence sequence

basedononanalysis based analysisofofananalignment alignment of multiple of multiple subtypes subtypes of a of a particular particular prostate prostate antigen. antigen.

Nucleic acidsequences Nucleic acid sequences that that encode encode a consensus a consensus polypeptide polypeptide sequence sequence may be prepared. may be prepared.

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Feb 2023 Vaccinescomprising Vaccines comprising proteins proteins thatthat comprise comprise consensus consensus sequences sequences and/or acid and/or nucleic nucleic acid moleculesthat molecules thatencode encode such such proteins proteins can can be used be used to induce to induce broad broad immunity immunity against aagainst a particular prostate particular prostate antigen. antigen. f. Electroporation f. Electroporation 2023200517 01 "Electroporation," "electro-permeabilization," "Electroporation," "electro-permeabilization," or "electro-kinetic or "electro-kinetic enhancement" enhancement"

("EP")asasused ("EP") usedinterchangeably interchangeably herein herein means means theofuse the use of a transmembrane a transmembrane electric electric field field pulse pulse to induce to microscopicpathways induce microscopic pathways (pores) (pores) in a in a bio-membrane; bio-membrane; their presence their presence allows allows biomoleculessuch biomolecules such as as plasmids, plasmids, oligonucleotides, oligonucleotides, siRNA, siRNA, drugs, drugs, ions, ions, and andtowater water pass to pass from from one side one side of ofthe the cellular cellular membrane membrane to the to the other. other.

g. Fragment g. Fragment "Fragment"as as "Fragment" used used herein herein with with respect respect to nucleic to nucleic acid acid sequences sequences means means a a nucleic nucleic

acid sequence acid sequenceorora aportion portionthereof, thereof,that thatencodes encodes a polypeptide a polypeptide capable capable of eliciting of eliciting an immune an immune

responseininaamammal response mammalthatthat cross cross reacts reacts withwith a full a full length length prostate prostate antigen. antigen. The fragments The fragments can can be DNA be DNA fragments fragments selected selected from from at least at least onetheofvarious one of the various nucleotide nucleotide sequences sequences that that encode encode the consensus the consensus amino acid sequences amino acid sequences and and constructs constructscomprising comprising such suchsequences. sequences. DNA DNA

fragmentscan fragments cancomprise comprise coding coding sequences sequences forimmunoglobulin for the the immunoglobulin leader leader such such as IgE as IgE or IgG or IgG sequences.DNADNA sequences. fragments fragments can encode can encode the protein the protein fragments fragments set forth set forth below. below. "Fragment"with "Fragment" with respect respect to to polypeptide polypeptide sequences sequences means means a polypeptide a polypeptide capable capable of of eliciting an eliciting an immune response immune response in ainmammal a mammal that cross that cross reactsreacts with awith a prostate prostate antigen, antigen,

including, e.g. including, e.g.PSA, PSA,PSMA, STEAPandand PSMA, STEAP PSCA. PSCA.

The human The humanPSA PSA sequence sequence is is about261 about 261amino amino acids.Fragments acids. Fragments of of PSA PSA consensus consensus

antigen 11may antigen comprise at may comprise at least least90%, 90%,91%, 91%, 92%, 92%, 93%, 94%, 95%, 93%, 94%, 95%,96%, 96%,97%, 97%, 98%, 98%, or or 99%99%

of SEQ of ID NO:2, SEQ ID NO:2,and andpreferably preferably 98% 99%,provided 98%oror99%, providedthe the fragments fragments include include one one or or more of more of

aminoacids amino acids69, 69,78,78,80,80,82,82,102, 102,110, 110, 137, 137, 139,139, 165,165, 189,189, 203,203, 220, 220, 232248. 232 and andFragments 248. Fragments of PSA of consensus PSA consensus antigen antigen 1 may1 may comprise comprise 255,257, 255, 256, 256, 257, 258, 259258, 259amino or 260 or 260 amino acids of acids of SEQIDIDNO:2, SEQ NO:2,but butpreferably preferably 256 256 amino aminoacids acids or or more. FragmentsofofPSA more. Fragments PSAconsensus consensus antigen 22 may antigen comprise at may comprise at least least90%, 90%,91%, 91%, 92%, 92%, 93%, 94%,95%, 93%, 94%, 95%,96%, 96%,97%, 97%, 98%, 98%, or or 99%99%

of SEQ of ID NO:4, SEQ ID NO:4,and andpreferably preferably 98% 99%,provided 98%oror99%, providedthe the fragments fragments include include one one or or more of more of

aminoacids amino acids21, 21,86,86,127, 127,129, 129, 154, 154, 156,156, 182,182, 195,195, 206,206, 218, 218, 220, 220, 237, 255, 237, 249, 249,265, 255,271 265, and271 and 275. All 275. Allsuch suchfragments fragments of PSA of PSA consensus consensus antigenantigen 2 may 2 may also also optionally optionally exclude exclude amino amino acids 1-17. acids 1-17. In In some some embodiments, fragments of embodiments, fragments of PSA PSAconsensus consensusantigen antigen 22 may mayoptionally optionally compriseone comprise oneorormore more of amino of amino acidsacids 1-17ofand 1-17 and theof the amino amino acids acids from from amino amino acid 18 toacid 18 to aminoacid amino acid278, 278,fragments fragments of PSA of PSA consensus consensus antigenantigen 2 may 2 may also also comprise comprise 255, 256, 255, 257, 256, 257, 258, 259 258, 259oror260 260amino amino acids acids of SEQ of SEQ ID NO:4, ID NO:4, but preferably but preferably 274 274 amino amino acids acids or more. or more.

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Feb 2023 The human The humanPSMA PSMA sequence sequence is about is about 749-750 749-750 amino amino acids. acids. Fragments Fragments of PSMA of PSMA

consensus antigen consensus antigen 11 may may comprise compriseat least 90%, 91%, at least 90%,92%, 93%,92%, 91%, 94%, 93%, 95%, 96%, 94%,97%,95%, 96%, 97%, 98%, or 98%, or 99% 99%ofofSEQ SEQIDIDNO:6, NO:6, and and preferably98% preferably 98%or or 99%, 99%, provided provided thefragments the fragmentsinclude include one or one or more moreofofamino amino acids acids 14, 32, 14, 15, 15, 47, 32, 47, 58, 58, 79,111, 79, 111, 157, 157, 223, 223, 320, 320, 350, 499, 350, 475, 475, 569, 499, 569, 2023200517 01 613, 624, 613, 624, 653, 653, 660, 660,663, 663,733 733and and734. 734.Fragments Fragments of ofPSMA consensusantigen PSMA consensus antigen 11 may may comprise745, comprise 745,746, 746, 747, 747, 748748 or 749 or 749 amino amino acids acids of SEQof IDSEQ NO:6,ID NO:6, but but preferably preferably 735 amino 735 amino acidsormore. acids or more. Fragments Fragments of PSMA consensusantigen PSMA consensus antigen2 2may maycomprise comprise atatleast least 90%, 90%,91%, 91%,

92%,93%,94%,95%,96%,97%,98%, 92%, or 99% 93%, 94%, 95%, 96%, 97%, 98%, or 99% of SEQ of SEQ ID NO:8, ID NO:8, and preferably and preferably 98% 98% or or 99%,provided 99%, providedthethe fragments fragments include include onemore one or or more of acids of amino amino21, acids 31, 21, 32, 31, 49, 32, 64, 49, 75, 64, 96, 75, 96, 128, 174, 128, 174, 240, 240,337, 337,367, 367,492, 492,516, 516, 565, 565, 586, 586, 630, 630, 641,641, 670,670, 677, 677, 680, 680, 750,751. 750, and andAll 751.suchAll such fragmentsofofPSA fragments PSA consensus consensus antigen antigen 2 may2also mayoptionally also optionally exclude exclude amino amino acids acids 1-16. In 1-16. In some embodiments, some embodiments,fragments fragmentsofofPSA PSA consensus consensus antigen2 2may antigen may optionallycomprise optionally compriseone oneoror moreofofamino more amino acids acids 1-171-17 andthe and of ofamino the amino acids acids fromacid from amino amino acid 18 to 18 acid amino to amino 767, acid 767, fragments of fragments of PSMA consensusantigen PSMA consensus antigen2 2may mayalso alsocomprise comprise762, 762, 763, 763, 764, 764, 765 765 or or 766 amino 766 amino

acids of acids of SEQ SEQIDID NO:8, NO:8, but but preferably preferably 752 acids 752 amino aminooracids more.or more. The human The humanSTEAP STEAP sequence sequence is about is about 339339 amino amino acids.Consensus acids. Consensus STEAP STEAP sequences sequences

maycomprise may comprise amino amino acid acid sequences sequences forimmunoglobulin for the the immunoglobulin leader leader such such as IgE as IgE or IgG. or IgG. ConsensusSTEAP Consensus STEAP antigen antigen 2 contains 2 contains an 18acid an 18 amino amino acidsequence leader leader sequence in place ofinthe place of the methionine at methionine at position position1.1.Fragments Fragments of ofPSMA consensus antigen PSMA consensus antigen 22 may may comprise compriseaa leader leader sequence and sequence and at at least least90%, 90%,91%, 91%, 92%, 92%, 9%,9%, 95%,96%, 93%, 94%, 95%, 96%, 97%, 97%, 98%, 98%, or or 99%99% of amino of amino

acids 18-356 acids 18-356 of SEQ ID NO:12, SEQ ID NO:12,and andpreferably preferably 98% 98%oror 99%. 99%.Fragments Fragments of of PSMA PSMA consensus consensus

antigen 11may antigen maycomprise comprise amino amino acidsacids 1-350,1-350, 1-351,1-351, 1-352, 1-352, 1-353,or1-354 1-353, 1-354 1-355 or of 1-355 SEQ ID of SEQ ID NO:12. NO:12.

The human The humanPSCA PSCA sequence sequence is is about about 114114 amino amino acids.Consensus acids. Consensus STEAP STEAP sequences sequences

maycomprise may comprise amino amino acid acid sequences sequences forimmunoglobulin for the the immunoglobulin leader leader such such as IgE as IgE or IgG. or IgG. ConsensusPSCA Consensus PSCA antigen antigen contains contains an 18 an 18 acid amino amino acidsequence leader leader sequence in the in place of place of the methionine at methionine at position position1.1.Fragments Fragments of ofPSCA consensus antigen PSCA consensus antigen may comprise aa leader may comprise leader sequence and sequence and at at least least90%, 90%,91%, 91%, 92%, 92%, 9%,9%, 95%,96%, 93%, 94%, 95%, 96%, 97%, 97%, 98%, 98%, or or 99%99% of amino of amino

acids 18-131 acids 18-131 of SEQ ID NO:14, SEQ ID NO:14,and andpreferably preferably 98% 98%oror 99%,. 99%,.Fragments FragmentsofofPSMA PSMA consensusantigen consensus antigen 1 may 1 may comprise comprise amino amino acids 1-125, acids 1-125, 1-126, 1-128, 1-126, 1-127, 1-127,1-129 1-128, 1-129ofor or 1-130 1-130 of SEQ ID SEQ ID NO:14. NO:14. h. Genetic h. Geneticconstruct construct As usedherein, As used herein,the theterm term"genetic "genetic construct" construct" refers refers to to thethe DNADNA or RNAormolecules RNA molecules that that comprisea anucleotide comprise nucleotide sequence sequence which which encodes encodes a protein. a protein. The sequence The coding coding sequence includes includes

-15- initiation and initiation and termination signalsoperably termination signals operably linked linked to to regulatory regulatory elements elements including including a promoter a promoter and polyadenylation and polyadenylation signal signal capable capable of directing of directing expression expression in cells in the the cells of the the individual of individual to to

2023200517 01 Feb whom whom thethe nucleic nucleic acid acid molecule molecule is administered. is administered. Asherein, As used used herein, the termthe term "expressible "expressible form" form" refers to refers to gene constructs that gene constructs thatcontain containthe thenecessary necessaryregulatory regulatory elements elements operable operable linkedlinked to a to a codingsequence coding sequence that that encodes encodes a protein a protein suchsuch that that whenwhen present present in theincell theofcell theofindividual, the individual, the coding the codingsequence sequence will will be be expressed. expressed.

i. Homology i. Homology Homologyofofmultiple Homology multiple sequence sequencealignments alignmentsand andphylogram phylogramwere weregenerated generatedusing using ClustalW,a ageneral ClustalW, generalpurpose purpose multiple multiple sequence sequence alignment alignment programprogram forproteins. for DNA or DNA or proteins. j. Identical j. Identical "Identical" or "Identical" or "identity" "identity" as as used used herein hereinininthe thecontext contextofoftwo twoorormore more nucleic nucleic acids acids or or polypeptidesequences, polypeptide sequences, means means that that the sequences the sequences have ahave a specified specified percentage percentage of residues of residues that that are the are the same overa aspecified same over specifiedregion. region.TheThe percentage percentage can can be be calculated calculated by optimally by optimally aligningaligning

the two the sequences,comparing two sequences, comparing the sequences the two two sequences over over the the specified specified region, region, determining determining the the numberofofpositions number positions at at which which thethe identical identical residue residue occurs occurs in both in both sequences sequences to yield to yield the the numberofofmatched number matched positions, positions, dividing dividing the number the number of matched of matched positions positions by the by the total total number number of positions of in the positions in the specified specified region, region, and andmultiplying multiplyingthethe resultbyby result 100100 to yield to yield thethe percentage percentage

of sequence of identity. InIncases sequence identity. caseswhere where the the two two sequences sequences are ofare of different different lengths lengths or theor the alignmentproduces alignment produces oneone or more or more staggered staggered endstheand ends and the specified specified region region of comparison of comparison

includes only includes onlya asingle singlesequence, sequence,thethe residues residues of of single single sequence sequence are included are included in thein the denominator but denominator but not not the the numerator numerator of ofthe thecalculation. When calculation. Whencomparing comparing DNA andRNA, DNA and RNA, thymine(T) thymine (T)andand uracil(U)(U) uracil cancan be be considered considered equivalent. equivalent. Identity Identity can becan be performed performed manually manually

or by or by using using aacomputer computer sequence sequence algorithm algorithm such such as asBLAST or BLAST BLAST or BLAST 2.0. 2.0.

k. Immune k. ImmuneResponse Response "Immune "Immune response" response" as used as used herein herein meansmeans the activation the activation of a host's of a host's immune immune system, system, e.g., that e.g., that of ofaamammal, mammal, ininresponse responseto to theintroduction the introduction of of antigen antigen suchsuch as a as a prostate prostate consensus consensus

antigen. The antigen. Theimmune immune response response caninbe can be theinform the of form of a cellular a cellular orhumoral or humoral response, response, or both.or both. 1. Nucleic 1. Nucleic Acid Acid "Nucleicacid" "Nucleic acid"oror"oligonucleotide" "oligonucleotide" or "polynucleotide" or "polynucleotide" as herein as used used herein means means at leastat least twonucleotides two nucleotidescovalently covalently linked linked together. together. The The depiction depiction of a single of a single strand strand also defines also defines the the sequenceofofthe sequence thecomplementary complementary strand. strand. Thus, Thus, a nucleic a nucleic acidencompasses acid also also encompasses the the complementary complementary strand strand of aof a depicted depicted single single strand. strand. Many Many variants variants of a nucleic of a nucleic acid canacid be can be usedfor used for the the same samepurpose purpose as as a given a given nucleic nucleic acid. acid. Thus, Thus, a nucleic a nucleic acid encompasses acid also also encompasses substantially identical substantially identical nucleic nucleic acids acids and andcomplements complements thereof. thereof. A single A single strandstrand provides provides a a

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Feb 2023 probethat probe that can canhybridize hybridizetotoa atarget targetsequence sequence under under stringent stringent hybridization hybridization conditions. conditions. Thus, Thus, a nucleic a acid also nucleic acid also encompasses encompasses a probe a probe thatthat hybridizes hybridizes underunder stringent stringent hybridization hybridization

conditions. conditions.

Nucleic acidscan Nucleic acids canbebesingle singlestranded stranded or or double double stranded, stranded, or contain or can can contain portions portions of of 2023200517 01 both double both doublestranded strandedandand single single stranded stranded sequence. sequence. The nucleic The nucleic acid acid can can be be DNA, DNA, both both genomic and genomic andcDNA, cDNA,RNA, RNA, or aorhybrid, a hybrid,where wherethethenucleic nucleic acid acid can can contain contain combinations combinations of of

deoxyribo-and deoxyribo- andribo-nucleotides, ribo-nucleotides, andand combinations combinations of bases of bases including including uracil,uracil, adenine, adenine,

thymine,cytosine, thymine, cytosine,guanine, guanine, inosine, inosine, xanthine xanthine hypoxanthine, hypoxanthine, isocytosine isocytosine and isoguanine. and isoguanine.

Nucleic acidscan Nucleic acids canbebeobtained obtained by by chemical chemical synthesis synthesis methods methods or by recombinant or by recombinant methods. methods.

m. Operably m. Operably Linked Linked "Operablylinked" "Operably linked" as as used used herein herein means means that that expression expression of a is of a gene gene is under under the control the control

of aa promoter of withwhich promoter with which it isspatially it is spatiallyconnected. connected. A promoter A promoter can becan be positioned positioned 5' 5' (upstream)oror3'3'(downstream) (upstream) (downstream)of aofgene a gene under under its control. its control. The distance The distance between between the the promoterand promoter anda gene a gene cancan be approximately be approximately the as the same same the as the distance distance betweenbetween that promoter that promoter

and the and the gene geneitit controls controls inin the the gene genefrom from which which the the promoter promoter is derived. is derived. As is As is known known in the in the art, variation art, variation in in this thisdistance distance can can be be accommodated without accommodated without loss loss of promoter of promoter function. function.

n. Promoter n. Promoter "Promoter"as asused "Promoter" used herein herein means means a synthetic a synthetic or naturally-derived or naturally-derived molecule molecule which iswhich is capableofofconferring, capable conferring,activating activatingororenhancing enhancing expression expression of a of a nucleic nucleic acid acid in a in a cell. cell. A A promotercan promoter cancomprise comprise one one or more or more specific specific transcriptional transcriptional regulatory regulatory sequences sequences to to further further enhanceexpression enhance expression and/or and/or to alter to alter thethe spatialexpression spatial expression and/or and/or temporal temporal expression expression of of same. same. A promoter A promotercancan also also comprise comprise distal distal enhancer enhancer or repressor or repressor elements, elements, which which can can be as be located located as muchasasseveral much severalthousand thousand basebase pairs pairs fromfrom the start the start sitesite of transcription. of transcription. A promoter A promoter can becan be derived from derived fromsources sources including including viral, viral, bacterial,fungal, bacterial, fungal,plants, plants,insects, insects,andand animals. animals. A A promotercan promoter canregulate regulatethethe expression expression of aof a gene gene component component constitutively, constitutively, or differentially or differentially

with respect with respecttoto cell, cell, the the tissue tissue or or organ in which organ in expression which expression occurs occurs or,or, with with respect respect to the to the

developmental developmental stage stage at at which which expression expression occurs, occurs, or in or in response response to external to external stimuli stimuli such assuch as physiologicalstresses, physiological stresses, pathogens, pathogens,metal metal ions, ions, or or inducing inducing agents. agents. Representative Representative examples examples of of promotersinclude promoters include thethe bacteriophage bacteriophage T7 promoter, T7 promoter, bacteriophage bacteriophage T3 promoter, T3 promoter, SP6 SP6 promoter, promoter, lac operator-promoter, lac operator-promoter,tac promoter, tac SV40 promoter, SV40late promoter, late SV40 promoter, SV40early earlypromoter, RSV-LTR promoter, RSV-LTR

promoter, CMV promoter, CMV IEIE promoter,SV40 promoter, SV40 earlypromoter early promoter ororSV40 SV40 latepromoter late promoterand andthe theCMV CMVIE IE promoter. promoter.

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Feb 2023 o. Stringent 0. StringentHybridization HybridizationConditions Conditions "Stringent hybridization "Stringent hybridizationconditions" conditions" as as used used herein herein means means conditions conditions underawhich under which a first nucleic first nucleic acid acid sequence (e.g., probe) sequence (e.g., probe) will willhybridize hybridizetotoa asecond second nucleic nucleic acid acid sequence sequence (e.g., (e.g., target), such target), as in such as in aa complex mixture complex mixture of of nucleic nucleic acids. acids. Stringent Stringent conditions conditions are sequence are sequence-

2023200517 01 dependentand dependent and will will bebe different different in in differentcircumstances. different circumstances. Stringent Stringent conditions conditions can becan be selected to selected to be be about about5-10°C 5-10°C lower lower thanthan the the thermal thermal melting melting point point (Tm) (Tm) for thefor the specific specific

sequenceatata adefined sequence definedionic ionicstrength strength pH. pH. The The T canTm be can the be the temperature temperature (underionic (under defined defined ionic

strength, pH, strength, pH, and andnucleic nucleicconcentration) concentration) at at which which 50% 50% of theofprobes the probes complementary complementary to the to the target hybridize target to the hybridize to the target target sequence sequenceatatequilibrium equilibrium(as(as thethe target target sequences sequences are are present present in in excess, at excess, at T, Tm,50%50% of the of the probes probes are occupied are occupied at equilibrium). at equilibrium). Stringent Stringent conditions conditions can be can be

those in those in which whichthe thesalt saltconcentration concentrationis isless lessthan thanabout about 1.01.0 M sodium M sodium ion, ion, such such as about as about 0.01- 0.01 1.0 M 1.0 sodium M sodium ionion concentration concentration (or (or other other salts) salts) at pH at pH 7.0 7.0 to 8.3 to 8.3 and and the temperature the temperature is at is at least least

about 30°C about 30°Cforforshort shortprobes probes (e.g.,about (e.g., about 10-50 10-50 nucleotides) nucleotides) andleast and at at least about about 60°C 60°C for for long long probes(e.g., probes (e.g., greater greater than than about about5050nucleotides). nucleotides).Stringent Stringent conditions conditions can also can also be achieved be achieved

with the with the addition additionofofdestabilizing destabilizingagents agentssuch such as as formamide. formamide. For selective For selective or specific or specific

hybridization, aapositive hybridization, positive signal signalcan canbebeatatleast least22 toto 1010times timesbackground background hybridization. hybridization.

Exemplary Exemplary stringent stringent hybridization hybridization conditions conditions include include the following: the following: 50% formamide, 50% formamide, 5x 5x SSC, and SSC, and 1% 1%SDS, SDS,incubating incubatingatat 42°C, 42°C, or, or, 5x 5x SSC, 1%SDS, SSC, 1% SDS,incubating at 65°C, incubating at 65°C, with wash wash

in 0.2x in 0.2x SSC, SSC, and and 0.1% SDS atat 65°C. 0.1% SDS 65°C. p. Substantially p. Substantially Complementary Complementary "Substantially complementary" "Substantially complementary" as used as used herein herein means means that a sequence that a first first sequence is at is at least least 60%, 65%, 60%, 65%,70%, 70%,75%, 75%, 80%, 80%, 85%,,9% 85%, 90%, 95%, 97%,97%, 98% 98% or 99%oridentical 99% identical to the to the complement complement

of aa second of sequence second sequence over over a region a region of 9, of 8, 8, 9, 10,10, 11,11, 12,12, 13, 13, 14, 14, 15,15, 16,16, 17, 17, 18, 18, 19, 19, 20,20, 21,21, 22, 22,

23,24,25,30,35,40,45,50,55,60,65,70,75,80,85,90,95,100,180,270,360,450,540, 23, 24, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 180, 270, 360, 450, 540,

630,720,810,900,990,1080,1170,1260,1350,1440,1530,1620,1710,1800,1890,1980, 630, 720, 810, 900, 990, 1080, 1170, 1260, 1350, 1440, 1530, 1620, 1710, 1800, 1890, 1980,

2070orormore 2070 more nucleotides nucleotides or amino or amino acids, acids, or that or that the the two sequences two sequences hybridize hybridize under stringent under stringent

hybridizationconditions. hybridization conditions. q. Substantially q. Substantially Identical Identical "Substantially identical" "Substantially identical"asasused usedherein hereinmeans means thatthat a first a first andand second second sequence sequence are atare at least 60%, least 60%, 65%, 65%, 70%, 75%,80%, 70%, 75%, 80%,85%, 85%, 90%, 90%, 95%, 95%, 97%, 97%, 98% 98% or 99% or 99% identical identical overover a region a region

of8,9, of 8, 9, 10, 10, 11,12, 11, 12, 13,14, 13, 14, 15,16, 15, 16, 17, 17, 18,19,20,21,22,23,24,25,30,35,40,45,50,55,60, 18, 19, 20, 21, 22, 23, 24, 25, 30, 35, 40, 45, 50, 55, 60,

65,70,75,80,85,90,95,100,180,270,360,450,540,630,720,810,900,990,1080,1170, 65, 70, 75, 80, 85, 90, 95, 100, 180, 270, 360, 450, 540, 630, 720, 810, 900, 990, 1080, 1170,

1260, 1350, 1260, 1350,1440, 1440,1530, 1530, 1620, 1620, 1710, 1710, 1800,1800, 1890, 1890, 1980, 1980, 2070 or2070 more or more nucleotides nucleotides or amino or amino

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Feb 2023 acids, or acids, or with respect to with respect to nucleic nucleic acids, acids, ifif the the first firstsequence is substantially sequence is substantially complementary complementary to to the complement the complement of the of the second second sequence. sequence.

r. Subtype r. SubtypeororSerotype Serotype "Subtype"oror"serotype": "Subtype" "serotype": as as used used herein, herein, interchangeably, interchangeably, and and in in reference reference to prostate to prostate

2023200517 01 cancer antigens, cancer antigens,means means genetic genetic variants variants of aofprostate a prostate cancer cancer antigen antigen such such thatsubtype that one one subtype (or (or variant) is variant) is recognized byananimmune recognized by immune system system apart apart from afrom a different different subtype. subtype.

s. Variant S. Variant "Variant"used "Variant" usedherein herein with with respect respect tonucleic to a a nucleic acid acid means means (i) a(i) a portion portion or fragment or fragment of of a referenced a referenced nucleotide nucleotidesequence; sequence; (ii)(ii) thethecomplement complement of a referenced of a referenced nucleotide nucleotide sequence sequence or or portion thereof; portion thereof; (iii) (iii) aa nucleic nucleic acid acid that that is issubstantially substantially identical identical to to aareferenced referenced nucleic acid nucleic acid

or the or the complement thereof; complement thereof; or (iv) or (iv) a nucleic a nucleic acid acid that that hybridizes hybridizes under under stringent stringent conditions conditions to to the referenced the nucleicacid, referenced nucleic acid,complement complement thereof, thereof, or a or a sequences sequences substantially substantially identical identical

thereto. thereto.

"Variant"with "Variant" withrespect respecttotoa apeptide peptideor or polypeptide polypeptide thatthat differs differs in in amino amino acid acid sequence sequence

by the by the insertion, insertion, deletion, deletion, or or conservative conservativesubstitution substitutionofofamino amino acids, acids, butbut retain retain at at leastoneone least

biological activity. biological activity. Variant canalso Variant can alsomean mean a protein a protein with with an amino an amino acid acid sequence sequence that isthat is substantially identical substantially identical to to aa referenced proteinwith referenced protein withananamino amino acidacid sequence sequence that that retains retains at least at least

one biological one biologicalactivity. activity. AAconservative conservative substitution substitution of amino of an an amino acid,acid, i.e.,i.e., replacing replacing an amino an amino

acid with acid with aa different different amino aminoacid acidof of similarproperties similar properties (e.g.,hydrophilicity, (e.g., hydrophilicity,degree degree andand

distribution of distribution of charged chargedregions) regions)isisrecognized recognizedin in thethe artart asas typicallyinvolving typically involving a minor a minor change. change.

Theseminor These minor changes changes can can be identified, be identified, in part, in part, by considering by considering the hydropathic the hydropathic index index of of aminoacids, amino acids,asasunderstood understoodin in thethe art.Kyte art. Kyte et al.,J.J.Mol. et al., Mol.Biol. Biol.157:105-132 157:105-132 (1982). (1982). The The hydropathicindex hydropathic index of of an an amino amino acidacid is based is based on a on a consideration consideration of itsof its hydrophobicity hydrophobicity and and charge. ItIt is charge. is known known inin theartartthat the thatamino amino acids acids of of similar similar hydropathic hydropathic indexes indexes can can be be substituted and substituted andstill still retains retains protein protein function. In one function. In oneaspect, aspect,amino amino acids acids having having hydropathic hydropathic

indexesofof±2 2are indexes aresubstituted. substituted.TheThe hydrophilicity hydrophilicity of amino of amino acids acids can be can also also betoused used to reveal reveal

that would substitutions that substitutions resultininproteins would result proteinsretaining biologicalfunction. retainingbiological function.A consideration A consideration of of the hydrophilicity the ofamino hydrophilicity of amino acids acids in in thethe context context of of a peptide a peptide permits permits calculation calculation of of the the greatest local greatest local average averagehydrophilicity hydrophilicityof of thatpeptide, that peptide,a auseful usefulmeasure measure thatthat has has beenbeen reported reported

to correlate to correlate well with antigenicity well with antigenicityand andimmunogenicity. immunogenicity. U.S. Patent U.S. Patent No. 4,554,101, No. 4,554,101,

incorporatedfully incorporated fullyherein hereinbybyreference. reference.Substitution Substitution of amino of amino acidsacids having having similar similar

hydrophilicityvalues hydrophilicity valuescancanresult resultininpeptides peptidesretaining retainingbiological biological activity,forforexample activity, example immunogenicity, immunogenicity, as understood as is is understood in the in the art.art. Substitutions Substitutions can can be performed be performed with acids with amino amino acids havinghydrophilicity having hydrophilicityvalues values within within ±2 of2 each of each other. other. Both Both the hydrophobicity the hydrophobicity index index and the and the

-19- valueofofamino hydrophilicityvalue hydrophilicity amino acids acids are are influenced influenced by particular by the the particular side side chainchain of that of that aminoamino acid. Consistent acid. Consistentwith withthat thatobservation, observation, amino amino acid acid substitutions substitutions that that are compatible are compatible with with

2023200517 01 Feb biological function biological functionare areunderstood understoodto to depend depend on relative on the the relative similarity similarity of the of the amino amino acids, acids, and and particularly the particularly the side side chains chains of ofthose thoseamino aminoacids, acids,as asrevealed revealed by by the the hydrophobicity, hydrophobicity,

hydrophilicity, charge, hydrophilicity, charge,size, size, and andother otherproperties. properties. t. Vector t. Vector "Vector"asasused "Vector" usedherein hereinmeans means a nucleic a nucleic acidacid sequence sequence containing containing an origin an origin of of replication. AAvector replication. vectorcan canbebe a vector,bacteriophage, a vector, bacteriophage, bacterial bacterial artificialchromosome artificial chromosome or or yeast yeast artificial chromosome. artificial A vector chromosome. A vector can can be a be DNAa or DNARNA or RNAAvector. vector. A vector vector can can be a self- be a self replicating extrachromosomal replicating extrachromosomal vector, vector, and preferably, and preferably, is a is DNAa plasmid. DNA plasmid.

2. Consensus 2. Consensusprostate prostateantigens antigens Providedherein Provided hereinareareconsensus consensus antigens antigens capable capable of eliciting of eliciting an immune an immune response response in a in a mammal mammal against against a prostate a prostate antigen. antigen. The consensus The consensus antigenantigen can comprise can comprise epitopes epitopes that make that make themparticularly them particularlyeffective effectiveasasimmunogens immunogens against against prostate prostate cancercancer cellsbecan cells can be induced. induced. The The consensusprostate consensus prostateantigen antigen cancan comprise comprise the full the full length length translation translation product, product, a variant a variant thereof, thereof,

a fragment a fragmentthereof thereoforora acombination combination thereof. thereof.

Sevendifferent Seven differentconsensus consensus prostate prostate antigens antigens havehave been been designed. designed. Two of Two the of the consensus prostate consensus prostate antigens antigensare areconsensus consensusPSA PSA antigen antigen1 1(SEQ (SEQ ID ID NO:2) NO:2) and and consensus PSA consensus PSA

antigen 22 (SEQ antigen ID NO:4). (SEQ ID Twoofofthe NO:4). Two theconsensus consensusprostate prostate antigens antigens are areconsensus consensusPSMA PSMA

antigen 11 (SEQ antigen ID NO:6) (SEQ ID andconsensus NO:6) and consensusPSMA PSMA antigen antigen 2 (SEQ 2 (SEQ ID ID NO:8). NO:8). Two Two of the of the

consensus prostate consensus prostate antigens antigensare areconsensus consensusSTEAP STEAP antigen antigen 11 (SEQ (SEQ ID NO:10)and ID NO:10) andconsensus consensus STEAPantigen STEAP antigen2 2(SEQ (SEQIDID NO:12). NO:12). OneOne of the of the consensus consensus prostateantigens prostate antigensisis consensus consensus PSCAantigen PSCA antigen(SEQ (SEQIDID NO:14). NO:14). Proteins Proteins may may comprise comprise sequences sequences homologous homologous to to the the prostate antigens, prostate antigens, fragments fragmentsof of theprostate the prostateantigens antigens andand proteins proteins withwith sequences sequences homologous homologous

to fragments to fragmentsofofthe theprostate prostateantigens. antigens. Consensus PSA Consensus PSAantigen antigen1 1(SEQ (SEQIDIDNO:2) NO:2) is is about91% about 91% homologous homologous to human to human PSA PSA sequences, about 95% sequences, about homologoustotoM.M.fascicuaris 95% homologous fascicuarisPSA PSAand andabout about96% 96%homologous homologous to to M. M.

mulatta PSA. mulatta PSA. Consensus Consensus PSA PSA antigen antigen 1 differsfrom 1 differs fromhuman humanPSAPSA sequences sequences at amino at amino acids acids

69, 78, 69, 78, 80, 80, 82, 82, 102, 102, 110, 110, 137, 137,139, 139,165, 165,189, 189, 203, 203, 220, 220, 232 232 and and 248SEQofIDSEQ 248 of ID NO:2. NO:2. Consensus PSA Consensus PSAantigen antigen2 2(SEQ (SEQIDIDNO:4) NO:4) is is about90-91% about 90-91% homologous homologous to human to human

PSAsequences, PSA sequences, about 95%homologous about 95% homologousto to M.M. fascicuarisPSA fascicuaris PSA and and about95%95% about homologous homologous

to M. to mulattaPSA. M. mulatta PSA. Consensus Consensus PSA antigen PSA antigen 2 comprises 2 comprises a leaderatsequence a leader sequence at its N its N terminus. terminus.

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Feb 2023 ConsensusPSAPSA Consensus antigen antigen 2 also 2 also differs differs from from humanhuman PSA sequences PSA sequences at amino at amino acids acids 21, 86, 127,21, 86, 127, 129, 154, 129, 154, 156, 156, 182, 182,195, 195,206, 206,218, 218,220, 220, 237, 237, 249,249, 255,255, 265,265, 271275 271 and andof275 SEQ of ID SEQ NO:4.ID NO:4. Consensus PSMA Consensus PSMA antigen antigen 1 (SEQ 1 (SEQ ID ID NO:6) NO:6) is about is about 96%96% homologous homologous to human to human

PSMA PSMA sequences sequences andand about about 94% 94% homologous homologous to mulatta to M. M. mulatta PSMA. PSMA. Consensus Consensus PSMA PSMA 2023200517 01 antigen 11differs antigen differs from fromhuman humanPSMAPSMA sequences sequences at aminoatacids amino 14,acids 14,47, 15, 32, 15,58, 32,79,111, 47, 58, 79,111, 157, 223, 157, 223, 320, 320, 350, 475,499, 350,475, 499,569, 569, 613, 613, 624, 624, 653,653, 660,660, 663,663, 733734 733 and andof734 SEQ of ID SEQ NO:6.ID NO:6. Consensus PSMA Consensus PSMA antigen antigen 2 (SEQ 2 (SEQ ID ID NO:8) NO:8) is about is about 96%96% homologous homologous to human to human PSA PSA sequences and sequences and about about 94% 94%homologous homologousto to M.M.mulatta mulattaPSA. PSA. Consensus Consensus PSMAPSMA antigen antigen 2 2 comprisesa aleader comprises leadersequence sequence at itsN terminus. at its N terminus. Consensus Consensus PSMA2 antigen PSMA antigen 2 alsofrom also differs differs from humanPSAPSA human sequences sequences at amino at amino acids acids 21, 31, 21, 32, 31, 49, 32, 64, 49, 75, 64, 96, 75, 128,96, 128, 174, 174, 240, 240, 337, 367,337, 367, 492, 516, 492, 516, 565, 565,586, 586,630, 630,641, 641, 670, 670, 677, 677, 680,680, 750,750, and and 751SEQofIDSEQ 751 of ID NO:8. NO:8. Consensus STEAP Consensus STEAP antigen1 (SEQ antigen 1 (SEQ ID ID NO:10) NO:10) is about is about 94%94% homologous homologous to some to some

humanSTEAP human STEAP sequences sequences and and about about 99%99% homologous homologous to other to other human human STEAPSTEAP sequences. sequences.

Consensus STEAP Consensus STEAP antigen antigen 1 (SEQ 1 (SEQ ID ID NO:10) NO:10) is alsoabout is also about94% 94% homologous homologous to mulatta to M. M. mulatta PSMA. PSMA. Consensus STEAP Consensus STEAP antigen2 2(SEQ antigen (SEQ ID ID NO:12) NO:12) is about is about 88%88% homologous homologous to some to some

humanSTEAP human STEAP sequences sequences and and about about 94%94% homologous homologous to other to other human human STEAPSTEAP sequences. sequences.

Consensus STEAP Consensus STEAP antigen antigen 2 2 (SEQ (SEQ ID ID NO:12) NO:12) is alsoabout is also about94% 94% homologous homologous to mulatta to M. M. mulatta PSMA.Consensus PSMA. Consensus STEAP STEAP antigen antigen 2 comprises 2 comprises a leader a leader sequence sequence at itsN Nterminus. at its terminus. Consensus PSCA Consensus PSCA antigen(SEQ antigen (SEQ ID ID NO:14) NO:14) is about is about 87%87% homologous homologous to human to human

PSCA.Consensus PSCA. Consensus PSCA PSCA antigen antigen (SEQ(SEQ ID NO:14) ID NO:14) differs differs fromfrom human human PSCA PSCA by inclusion by inclusion of of a leader a leader sequence sequenceatatits its NNterminus. terminus. Proteins may Proteins have sequences may have sequences 98% 98%homologous homologousto to PSA PSA Consensus Consensus Antigen Antigen sequence sequence

1 (SEQ 1 ID NO:2), (SEQ ID NO:2), PSA PSAConsensus Consensus Antigen Antigen sequence sequence 2 (SEQ 2 (SEQ ID NO:4), ID NO:4), PSMAPSMA Consensus Consensus

Antigen sequence Antigen sequence 11 (SEQ (SEQIDIDNO:6), NO:6),PSMA PSMA Consensus Consensus Antigen Antigen sequence sequence 2 (SEQ 2 (SEQ ID NO:8), ID NO:8),

STEAPConsensus STEAP Consensus Antigen Antigen sequence sequence 1 (SEQ 1 (SEQ ID NO:10), ID NO:10), STEAP STEAP Consensus Consensus AntigenAntigen

sequence 22 (SEQ sequence (SEQID IDNO:12) NO:12)ororPSCA PSCA Consensus Consensus Antigen Antigen sequence sequence (SEQ(SEQ ID NO:14). ID NO:14).

Proteins may Proteins have sequences may have sequences 99% 99%homologous homologousto to PSA PSA Consensus Consensus Antigen Antigen sequence sequence

As noted above, As noted above, some embodimentscomprise some embodiments comprisea aleader leadersequence sequenceatat the the N terminus. In N terminus. In someembodiments, some embodiments, the leader the leader sequence sequence is an is IgEan IgE leader leader sequence sequence thatIDis NO:16. that is SEQ SEQ ID In NO:16. In

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Feb 2023 some embodiments some embodimentsofofthe theprotein protein sequences sequences provided provided herein, herein, SEQ ID NO:16 SEQ ID NO:16isis removed removed therefrom. Likewise, therefrom. Likewise, in in some some embodiments embodiments of the nucleic of the nucleic acid sequences acid sequences provided provided herein, herein, SEQIDIDNO:15 SEQ NO:15 (which (which encodes encodes SEQSEQ ID NO:16) ID NO:16) is removed is removed therefrom. therefrom.

Accordingly, some Accordingly, some embodiments embodiments relatedrelated proteinprotein that comprise that comprise a signalapeptide signal peptide linked linked

2023200517 01 to SEQ to ID NO:2, SEQ ID NO:2,SEQ SEQIDID NO:6, NO:6, or or SEQSEQ ID NO:10 ID NO:10 in place in place of the of the N terminalmethionine N terminal methionine set set

forth in forth in the the claim (the coding claim (the codingsequence sequenceof of thethe signal signal peptide peptide typically typically includes includes a stsart a stsart codon codon

encodingananN N encoding terminal terminal methionine). methionine). Some embodiments Some embodiments relate to relate to athat a protein protein that comprises comprises a a signal peptide signal peptidelinked linkedtoto amino aminoacid 19-131 acid ofof 19-131 SEQ SEQIDIDNO:14. NO:14. Some embodimentsrelated Some embodiments relatedtoto proteins that proteins that comprise comprisea asignal signalpeptide peptidelinked linked to to a protein a protein 98%98 % homologous homologous to NO:2 to SEQ ID SEQ ID NO:2 providedamino provided amino acids acids 69,69, 78,78, 80, 80, 82, 82, 102,102, 110,110, 137, 137, 139, 139, 165, 165, 189, 220, 189, 203, 203, 232 220,and232 248and of 248 of SEQIDID SEQ NO:2 NO:2 are conserved. are conserved. Some embodiments Some embodiments related to related proteinstothat proteins thata comprise comprise signal a signal peptide linked peptide linked totoa a protein 98 98% protein % homologous to SEQ homologous to ID NO:6 SEQ ID NO:6provided providedamino aminoacids acids14, 14,15, 15, 32, 47, 32, 47, 58, 58, 79, 79,111, 157, 223, 111, 157, 223, 320, 320,350, 350,475, 475,499, 499,569, 569, 613, 613, 624, 624, 653,653, 660,660, 663, 663, 733734 733 and and 734 of SEQ of ID NO:6 SEQ ID NO:6are areconserved. conserved. Some Some embodiments embodiments related related to to proteinsthat proteins that comprise comprise aa signal peptide signal peptidelinked linkedtoto a protein 98 % a protein 98%homologous homologous to to SEQ ID NO:10, SEQ ID NO:10,InIneach eachinstance instance in in whichthe which thesignal signalpeptide peptideisislinked linkedatatthe theN Nterminal terminal it it isislinked linkedininplace placeofofthe theN Nterminal terminal methioninesetsetforth methionine forthininthe theclaim claim(the (thecoding coding sequence sequence of signal of the the signal peptide peptide typically typically includes includes

a stsart a stsart codon encodinganan codon encoding N terminal N terminal methionine). methionine). Some embodiments Some embodiments relate to arelate to protein a protein that comprises that comprises a asignal signalpeptide peptidelinked linked to to linked linked to to a protein a protein 98%98homologous % homologous to aminotoacid amino acid 19-131 ofofSEQ 19-131 SEQID ID NO:14. NO:14. Some embodiments Some embodiments relate to arelate to that protein a protein that comprises comprises a signal a signal peptide linked peptide linked totolinked to to linked an an immunogenic immunogenicfragment fragmentofofSEQ SEQ ID ID NO:2 comprising amino NO:2 comprising amino acids corresponding acids correspondingto toat atleast least256 256amino amino acidacid residues residues of SEQ of SEQ ID provided ID NO:2, NO:2, provided amino amino acids 69, acids 69, 78, 78, 80, 80, 82, 82, 102, 102, 110, 110,137, 137,139, 139,165, 165,189, 189, 203, 203, 220, 220, 232 232 and of and 248 248 SEQofIDSEQ NO:2ID NO:2 are conserved. are conserved.Some Some embodiments embodiments relate relate to a protein to a protein that comprises that comprises a signala peptide signal peptide linked linked to linked to linkedtotoananimmunogenic immunogenic fragment fragment of of SEQ ID NO:6 SEQ ID NO:6comprising comprisingamino aminoacids acids correspondingto toatatleast corresponding least735 735amino amino acidacid residues residues of SEQ of SEQ ID NO:6, ID NO:6, providedprovided amino amino acids 14, acids 14, 32, 47, 15, 32, 15, 58, 79,111, 47, 58, 79,111, 157, 157,223, 223,320, 320,350, 475, 350,475, 499, 499, 569, 569, 613,613, 624,624, 653, 653, 660, 660, 663, 663, 733 733 and and 734 ofofSEQ 734 SEQID ID NO:6 NO:6 are conserved. are conserved. Some embodiments Some embodiments relate to a relate tothat protein a protein that acomprises comprises a signal peptide signal peptidelinked linkedtoto an an immunogenic immunogenic fragment fragmentof ofSEQ SEQ ID ID NO:10 comprisingamino NO:10 comprising aminoacids acids corresponding to corresponding to at atleast least333333 amino aminoacid residues acid of SEQ residues IDID of SEQ NO:10. NO:10.Someembodiments Some embodiments

relate to relate to aa protein protein that that comprises comprises a asignal signal peptide peptidelinked linkedtotolinked linkedto toprotein proteinthat thathashasa signal a signal peptide linked peptide linked totoananimmunogenic immunogenic fragment of amino fragment of amino acids acids 19-131 19-131 of of SEQ ID NO:14, SEQ ID NO:14, the the fragmentcomprising fragment comprising at least at least 110110 amino amino acid acid residues residues ofIDSEQ of SEQ ID NO:14. NO:14.

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Feb 2023 3. Genetic 3. GeneticSequences, Sequences,Constructs Constructs and and Plasmids Plasmids

Nucleic acidmolecules Nucleic acid molecules encoding encoding the consensus the consensus amino amino acid acid sequences sequences were generated were generated

to optimize to stability and optimize stability and expression expressionin inhumans. humans. CodonCodon selection selection was determined was determined based based upon, upon, inter alia, inter alia, an an effort effort to tominimize intramolecularinteractions minimize intramolecular interactionsandand secondary secondary structure structure formation formation

2023200517 01 as well as as using well as using codons codonswhich which result result in in improved improved expression. expression. Vaccines Vaccines may comprise may comprise one or one or morenucleic more nucleicacid acidsequences sequences thatthat encode encode onemore one or or of more the of the consensus consensus versionsversions of the of the immunogenic immunogenic proteins proteins selected selected from from this group this group of sequences of sequences generated generated to optimize to optimize stability stability

and expression and expressionininhumans. humans. Nucleic Nucleic acid sequences acid sequences incorporating incorporating coding sequence coding sequence for the IgEfor the IgE leader at leader at the the 5' 5' end of the end of the optimized, optimized,consensus consensus encoding encoding nucleic nucleic acid acid sequence sequence were were generatedwhich generated which encoded encoded proteins proteins having having theleader the IgE IgE leader sequence sequence at the Natterminus the N terminus of the of the consensusamino consensus amino acid acid sequence. sequence. In embodiments, In some some embodiments, theacid the nucleic nucleic acid that sequence sequence that encodes the encodes the IgE IgE leader leaderisisSEQ SEQ ID ID NO:15 NO:15

Nucleic acid sequences Nucleic acid sequences are areprovided providedwhich which encode encode PSA ConsensusAntigen PSA Consensus Antigensequence sequence 1 (protein 1 (proteinsequence sequenceSEQ SEQ ID ID NO:2; nucleic acid NO:2; nucleic acid sequence sequence SEQ ID NO:1), SEQ ID NO:1), PSA PSAConsensus Consensus Antigen sequence Antigen sequence 22 (protein (protein sequence sequence SEQ ID NO:4; SEQ ID NO:4;nucleic nucleic acid acid sequence sequence SEQ SEQIDIDNO:3), NO:3), PSMA PSMA Consensus Consensus Antigen Antigen sequence sequence 1 (protein 1 (protein sequence sequence SEQSEQ ID NO:6; ID NO:6; nucleic nucleic acidacid sequence having sequence having nucleotides nucleotides 1-2250 1-2250 of of SEQ ID NO:5), SEQ ID NO:5), PSMA PSMA Consensus Consensus Antigen Antigen sequence sequence

2 (protein 2 (proteinsequence sequence SEQ ID NO:8; SEQ ID nucleic acid NO:8; nucleic acid sequence sequence having having nucleotides nucleotides 1-2301 1-2301 of ofSEQ SEQ

ID NO:7), ID NO:7), STEAP STEAP Consensus Consensus Antigen Antigen sequence sequence 1 (protein 1 (protein sequence sequence SEQSEQ ID NO:10; ID NO:10; nucleic nucleic

acid sequence acid sequence SEQ IDNO:9), SEQ ID NO:9),STEAP STEAP Consensus Consensus Antigen Antigen sequence sequence 2 (protein 2 (protein sequence sequence SEQ SEQ

ID NO:12; ID NO:12;nucleic nucleic acid acid sequence sequence SEQ IDNO:11) SEQ ID NO:11)ororPSCA PSCA Consensus Consensus Antigen Antigen sequence sequence

(protein sequence (protein sequence SEQ ID NO:14; SEQ ID NO:14;nucleic nucleic acid acid sequence sequence SEQ SEQIDIDNO:13). NO:13).TheThe nucleicacid nucleic acid sequence SEQ sequence SEQIDIDNO:5 NO:5 which which encodes encodes PSMA PSMA Consensus Consensus Antigen Antigen sequence sequence 1 comprises, 1 comprises, in in addition to addition to PSMA PSMA encoding encoding nucleotides, nucleotides, an additional an additional 9 codons 9 codons (27 nucleotides) (27 nucleotides) immediately immediately

before the before the stop stopcodons codonswhich which encode encode the theHA Tag (SEQ HA Tag (SEQIDIDNO:32), NO:32),not notshown shownininSEQ SEQID ID

NO:6. The NO:6. The HA is HA Tag Tag is peptide peptide sequence sequence that corresponds that corresponds to an influenza to an influenza epitope epitope useful foruseful for

amongother among other things things detection detection of protein. of protein. expression expression using using commercially commercially available available anti-HA anti-HA

Tag antibodies. Tag antibodies. SEQ IDNO:5 SEQ ID NO:5encodes encodesSEQSEQ ID ID NO:6 NO:6 plusplus an additional9 amino an additional 9 amino acid acid

sequenceSEQ sequence SEQ ID NO:32 ID NO:32 linkedlinked to atN its to at its N terminus terminus to the to the C terminus C terminus ofNO:6. of SEQ ID SEQ In ID NO:6. In some embodiments, some embodiments,the thePSMA-1 PSMA-1 Consensus Consensus antigen antigen is encoded is encoded by by SEQSEQ ID NO:5 ID NO:5 and and comprisesa aproteins comprises proteinshaving having an an amino amino acid acid sequence sequence of SEQ of ID SEQ NO:6 ID NO:6 linked linked at its at its C C terminus terminus to the to theNN terminus terminusof ofSEQ SEQ ID ID NO:32. In some NO:32. In someembodiments, embodiments,the thePSMA-1 PSMA-1 Consensus Consensus antigen antigen

is encoded is encoded by by nucleotides nucleotides1-2250 1-2250 of ofSEQ SEQ ID ID NO:5 and comprises NO:5 and comprises aa proteins proteins having having an an amino amino

acid sequence acid sequence of SEQ ID NO:6. SEQ ID NO:6. The Thecoding codingsequence sequencehaving havingnucleotides nucleotides1-2250 1-2250ofofSEQ SEQID ID

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Feb 2023 NO:5 hasoneone NO:5 has or or more more stopstop codons codons at itsat3' itsend. 3' end. The nucleic The nucleic acid sequence acid sequence SEQ ID NO:7 SEQ ID NO:7

which encodes which encodes PSMA PSMA Consensus Consensus Antigen Antigen sequence sequence 2 comprises, 2 comprises, in additiontotonucleotides in addition nucleotides encodingthe encoding theIgE IgEsignal signallinked linked to to thethe PSMA, PSMA, protein protein plus plus an an additional additional 9 codons 9 codons (27 (27 nucleotides) immediately nucleotides) immediately before before the thestop codons stop codonswhich whichencode encodethe theHA HA Tag Tag (SEQ ID NO:32), (SEQ ID NO:32), 2023200517 01 not shown not in SEQ shown in SEQID IDNO:8. NO:8.SEQSEQ ID NO:7 ID NO:7 encodes encodes SEQ SEQ ID ID plus NO:8 NO:8an plus an additional additional 9 9 aminoacid amino acidsequence sequence SEQ SEQ ID NO:32 ID NO:32 linked linked to to Natterminus at its its N terminus to the C to the C terminus terminus of SEQ ID of SEQ ID NO:8. In some NO:8. In someembodiments, embodiments, thePSMA-2 the PSMA-2 Consensus Consensus antigen antigen is encoded is encoded by SEQ by SEQ ID NO:7 ID NO:7

and comprises and comprisesa proteins a proteins having having an amino an amino acid sequence acid sequence of SEQ of ID SEQ ID NO:8 NO:8 linked linked at its C at its C terminus to terminus to the theNNterminus terminusof ofSEQ SEQ ID ID NO:32. In some NO:32. In someembodiments, embodiments,the thePSMA-2 PSMA-2 Consensus antigen Consensus antigen is is encoded encoded by by nucleotides nucleotides 1-2301 1-2301 of ofSEQ SEQ ID ID NO:7 and comprises NO:7 and comprises aa proteins having proteins having an an amino amino acid acid sequence sequence of ofSEQ SEQ ID ID NO:8. Thecoding NO:8. The codingsequence sequencehaving having nucleotides1-2301 nucleotides 1-2301of of SEQSEQ ID NO:7 ID NO:7 has has one or one moreor more stop stopatcodons codons its 3' at its end. 3' end. Isolated nucleic Isolated nucleic acid acidmolecules moleculescancan encode encode proteins proteins that that have have sequences sequences 98% 98% homologoustotoPSA homologous PSAConsensus Consensus Antigen Antigen sequence sequence 1 (SEQ 1 (SEQ ID NO:2), ID NO:2), provided provided amino amino acids acids

69, 78, 69, 78, 80, 80, 82, 82, 102, 102, 110, 110, 137, 137,139, 139,165, 165,189, 189, 203, 203, 220, 220, 232 232 and and 248SEQofIDSEQ 248 of NO:2ID NO:2 are are conserved, PSA conserved, ConsensusAntigen PSA Consensus Antigensequence sequence2 2(SEQ (SEQ ID ID NO:4), NO:4), provided provided amino amino acids acids 21,21,

86, 127, 86, 127, 129, 129, 154, 154,156, 156,182, 182,195, 195,206, 206, 218, 218, 220, 220, 237,237, 249,249, 255, 255, 265, 265, 271275 271 and andof 275 SEQ of ID SEQ ID NO:4 are conserved, NO:4 are conserved, PSMA PSMA Consensus Consensus Antigen Antigen sequence sequence 1 (SEQ 1 (SEQ ID NO:6), ID NO:6), provided provided amino amino

acids 14, acids 14, 15, 15, 32, 32, 47, 47, 58, 58, 79, 79,111, 157, 223, 111, 157, 223,320, 320,350, 350,475, 475,499, 499, 569, 569, 613, 613, 624,624, 653,653, 660,660, 663, 663,

733 and 733 and 734 734 of of SEQ IDNO:6 SEQ ID NO:6are areconserved, conserved, PSMA PSMA Consensus Consensus Antigen Antigen sequence sequence 2 (SEQ 2 (SEQ ID ID NO:8), provided NO:8), provided amino amino acids acids 21, 32, 21, 31, 31, 49, 32, 64, 49, 75, 64, 96, 75, 128, 96, 128, 174, 337, 174, 240, 240, 367, 337, 492, 367,516, 492, 516, 565, 586, 565, 586, 630, 630, 641, 641,670, 670,677, 677,680, 750, 680, andand 750, 751751 of of SEQ NO:8 are SEQIDIDNO:8 areconserved, conserved,STEAP STEAP

Consensus Antigen Consensus Antigensequence sequence1 1(SEQ (SEQIDIDNO:10), NO:10),STEAP STEAP Consensus Consensus Antigen Antigen sequence sequence 2 2 (SEQ IDIDNO:12) (SEQ NO:12)ororPSCA PSCA Consensus Consensus Antigen Antigen sequence sequence (SEQ(SEQ ID NO:14). ID NO:14).

Isolated nucleic Isolated nucleic acid acidmolecules moleculescancan encode encode proteins proteins that that have have sequences sequences 99% 99% homologoustotoPSA homologous PSAConsensus Consensus Antigen Antigen sequence sequence 1 (SEQ 1 (SEQ ID NO:2), ID NO:2), provided provided amino amino acids acids

86, 127, 86, 127, 129, 129, 154, 154,156, 156,182, 182,195, 195, 206, 206, 218, 218, 220, 220, 237,237, 249,249, 255, 255, 265, 265, 271275 271 and andof 275 SEQ of ID SEQ ID NO:4 are conserved, NO:4 are conserved, PSMA PSMA Consensus Consensus Antigen Antigen sequence sequence 1 (SEQ 1 (SEQ ID NO:6), ID NO:6), provided provided amino amino

733 and 733 and 734 734 of of SEQ IDNO:6 SEQ ID NO:6are areconserved, conserved, PSMA PSMA Consensus Consensus Antigen Antigen sequence sequence 2 (SEQ 2 (SEQ ID ID NO:8), provided NO:8), provided amino amino acids acids 21, 32, 21, 31, 31, 49, 32, 64, 49, 75, 64, 96, 75, 128, 96, 128, 174, 337, 174, 240, 240, 367, 337, 492, 367,516, 492, 516, 565, 586, 565, 630, 641, 586, 630, 641,670, 670,677, 677,680, 750, 680, andand 750, of of 751751 SEQSEQIDIDNO:8 NO:8 are areconserved, conserved,STEAP STEAP

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Feb 2023 Consensus Antigen Consensus Antigensequence (SEQIDIDNO:10), sequence1 1(SEQ NO:10),STEAP STEAP Consensus Consensus Antigen Antigen sequence sequence 2 2 (SEQ IDIDNO:12) (SEQ NO:12)ororPSCA PSCA Consensus Consensus Antigen Antigen sequence sequence (SEQ(SEQ ID NO:14). ID NO:14).

Isolated nucleic Isolated nucleic acid acidmolecules moleculescancan encode encode proteins proteins that that have have sequences sequences 98% 98% homologoustoto the homologous the sequence sequence encoding encoding PSA PSAConsensus Consensus Antigen Antigen sequence sequence 1 (SEQ 1 (SEQ ID NO:1), ID NO:1),

2023200517 01 PSAConsensus PSA ConsensusAntigen Antigensequence sequence 2 (SEQ 2 (SEQ ID ID NO:3), NO:3), PSMA PSMA Consensus Consensus Antigen Antigen sequence sequence 1 1 (SEQ IDIDNO:5 (SEQ NO:5ororpreferably preferably nucleotides nucleotides 1-2250 1-2250 of of SEQ IDNO:5), SEQ ID NO:5),PSMA PSMA Consensus Consensus

Antigen sequence Antigen sequence 22 (SEQ (SEQIDIDNO:7 NO:7ororpreferably preferably nucleotides nucleotides 1-2301 1-2301 of of SEQ IDNO:7), SEQ ID NO:7), STEAPConsensus STEAP Consensus Antigen Antigen sequence sequence 1 (SEQ 1 (SEQ ID NO:9), ID NO:9), STEAP STEAP Consensus Consensus Antigen Antigen

sequence 22 (SEQ sequence (SEQID IDNO:11) NO:11)ororPSCA PSCA Consensus Consensus Antigen Antigen sequence sequence (SEQ(SEQ ID NO:13). ID NO:13).

Isolated nucleic Isolated nucleic acid acidmolecules moleculescancan encode encode proteins proteins that that have have sequences sequences 99% 99% homologoustotothe homologous the sequence sequence encoding encoding PSA PSAConsensus Consensus Antigen Antigen sequence sequence 1 (SEQ 1 (SEQ ID NO:1), ID NO:1),

PSAConsensus PSA ConsensusAntigen Antigensequence sequence 2 (SEQ 2 (SEQ ID ID NO:3), NO:3), PSMA PSMA Consensus Consensus Antigen Antigen sequence sequence 1 1 (SEQ IDIDNO:5 (SEQ NO:5ororpreferably preferably nucleotides nucleotides 1-2250 1-2250 of of SEQ IDNO:5), SEQ ID NO:5),PSMA PSMA Consensus Consensus

Antigen sequence Antigen sequence 22 (SEQ (SEQIDIDNO:7 NO:7Or Orpreferably preferably nucleotides nucleotides 1-2301 1-2301 of of SEQ IDNO:7), SEQ ID NO:7), STEAPConsensus STEAP Consensus Antigen Antigen sequence sequence 1 (SEQ 1 (SEQ ID NO:9), ID NO:9), STEAP STEAP Consensus Consensus Antigen Antigen

Isolated nucleic Isolated nucleic acid acidmolecules moleculescancan encode encode proteins proteins that that comprise comprise a leader a leader sequence sequence

at the at the N terminus. InInsome N terminus. some embodiments, embodiments, the nucleic the nucleic acid molecules acid molecules canthe can encode encode IgE the IgE leader sequence leader sequencethat thatisisSEQ SEQID ID NO:16. NO:16. In embodiments In some some embodiments isolatedacid isolated nucleic nucleic acid moleculescancanencode molecules encode proteins proteins thatthat comprise comprise a signal a signal peptide peptide linkedlinked to SEQtoIDSEQ NO:2,ID NO:2, SEQ SEQ ID NO:6, ID NO:6,ororSEQSEQ ID NO:10 ID NO:10 in place in place of the of the N terminal N terminal methionine methionine set forthset in forth in the the claim claim (the (the codingsequence coding sequenceof of thethe signal signal peptide peptide typically typically includes includes a stsart a stsart codon codon encoding encoding an N an N terminal methionine). terminal methionine).In In some some embodiments embodiments isolatedisolated nucleic nucleic acid molecules acid molecules can encodecan encode proteins that proteins that comprise comprisea asignal signalpeptide peptidelinked linked to to amino amino acidacid 19-131 19-131 of SEQofIDSEQ ID In NO:14. NO:14. In someembodiments some embodiments isolated isolated nucleic nucleic acid molecules acid molecules can proteins can encode encode proteins that comprise that comprise a a signal peptide signal peptidelinked linkedtoto a protein 98 % a protein homologous 98% homologous to toSEQ SEQ ID ID NO:2 provided amino NO:2 provided aminoacids acids 69, 78, 69, 78, 80, 80, 82, 82, 102, 102, 110, 110, 137, 137,139, 139,165, 165,189, 189, 203, 203, 220, 220, 232 232 and and 248SEQofIDSEQ 248 of NO:2ID NO:2 are are conserved.InInsome conserved. some embodiments embodiments isolated isolated nucleicnucleic acid molecules acid molecules canproteins can encode encodethat proteins that comprise aa signal comprise signalpeptide peptidelinked to to linked a protein 98 % a protein homologous 98% homologous totoSEQ SEQ ID ID NO:6 provided NO:6 provided

aminoacids amino acids14,14,15,15,32,32,47,47,58,58,79,111, 79,111, 157, 157, 223, 223, 320,320, 350,350, 475, 475, 499, 499, 569, 569, 613, 653, 613, 624, 624,660, 653, 660, 663, 733 663, 733 and and 734 734 of of SEQ ID NO:6 SEQ ID NO:6are areconserved. conserved. InInsome someembodiments embodiments isolatednucleic isolated nucleic acid molecules acid moleculescancanencode encode proteins proteins thatthat comprise comprise a signal a signal peptide peptide linkedlinked to a protein to a protein 98% 98 %

homologous homologous to to SEQSEQ ID NO:10. ID NO:10. In instance In instance in whichincoding whichsequence coding for sequence a signalfor a signal peptide is peptide is provides, the provides, the signal signal peptide peptideisislinked linkedtotothe thepeptide peptidesequence sequence in place in place of the of the N terminal N terminal

-25-

Feb 2023 methioninesetsetforth methionine forthininthe thesequences sequences shown shown (the (the coding coding sequence sequence of the of the signal signal peptidepeptide

typically includes typically includesaastsart stsart codon encoding codon encoding an an N terminal N terminal methionine). methionine). In someInembodiments some embodiments isolated nucleic isolated nucleic acid acid molecules moleculescancan encode encode proteins proteins that that comprise comprise a signal a signal peptide peptide linked linked to to linked to linked toa aprotein protein98 98% % homologous to amino homologous to acid 19-131 amino acid 19-131 of of SEQ IDNO:14. SEQ ID NO:14.InInsome some 2023200517 01 embodiments embodiments isolated isolated nucleic nucleic acidacid molecules molecules can encode can encode proteins proteins that comprise that comprise a signal a signal peptide linked peptide linked totolinked to to linked an an immunogenic immunogenicfragment fragmentofofSEQ SEQ ID ID NO:2 comprising amino NO:2 comprising amino acids corresponding acids correspondingto toat atleast least256 256amino amino acidacid residues residues of SEQ of SEQ ID provided ID NO:2, NO:2, provided amino amino acids 69, acids 69, 78, 78, 80, 80, 82, 82, 102, 102, 110, 110,137, 137,139, 139,165, 165,189, 189, 203, 203, 220, 220, 232 232 and of and 248 248 SEQofIDSEQ NO:2ID NO:2 are conserved. are conserved.InInsome some embodiments embodiments isolated isolated nucleicnucleic acid molecules acid molecules canproteins can encode encode proteins that comprise that comprise a asignal signalpeptide peptidelinked linked to to linked linked to to an an immunogenic immunogenic fragment fragment of NO:6 of SEQ ID SEQ ID NO:6 comprisingamino comprising amino acids acids corresponding corresponding to at to at least least 735 amino 735 amino acid residues acid residues of NO:6, of SEQ ID SEQ ID NO:6, providedamino provided amino acids acids 14, 14, 15, 15, 32, 32, 47, 47, 58, 58, 79, 79,111, 111, 157,157, 223,223, 320,320, 350,350, 475, 475, 499, 499, 569, 624, 569, 613, 613, 624, 653, 660, 653, 660, 663, 663, 733 733 and and 734 734 of ofSEQ ID NO:6 SEQ ID are conserved. NO:6 are conserved. SS In In some some embodiments embodimentsisolated isolated nucleic acid nucleic acid molecules moleculescancan encode encode proteins proteins that that comprise comprise a signal a signal peptide peptide linked linked to an to an immunogenicfragment immunogenic fragmentofofSEQ SEQID ID NO:10 NO:10 comprising comprising amino amino acids acids corresponding corresponding to at to at least least

333 amino 333 amino acid acid residues residues of of SEQ ID NO:10. SEQ ID NO:10. InInsome someembodiments embodiments isolatednucleic isolated nucleicacid acid moleculescancanencode molecules encode proteins proteins thatthat comprise comprise a signal a signal peptide peptide linkedlinked to linked to linked to protein to protein that that has aa signal has signal peptide peptide linked linkedtotoananimmunogenic immunogenic fragment fragment of acids of amino amino19-131 acids of19-131 SEQ IDof SEQ ID NO:14, thefragment NO:14, the fragment comprising comprising at least at least 110 110 aminoamino acid residues acid residues of SEQ of ID SEQ NO:14.ID NO:14.

Providedherein Provided hereinarearegenetic geneticconstructs constructs that that cancan comprise comprise a nucleic a nucleic acid acid sequence sequence that that encodesconsensus encodes consensus prostate prostate antigen antigen disclosed disclosed herein herein including including consensus consensus protein protein sequences, sequences,

sequenceshomologous sequences homologous to consensus to consensus protein protein sequences, sequences, fragments fragments of consensus of consensus protein protein sequences and sequences and sequences sequences homologous homologoustotofragments fragmentsofofconsensus consensus protein protein sequences. The sequences. The

genetic construct genetic constructcan canbebepresent presentininthethecell cellasasa afunctioning functioningextrachromosomal extrachromosomal molecule. molecule. The The genetic construct genetic constructcan canbebelinear linearminichromosome minichromosome including including centromere, centromere, telomers telomers or plasmids or plasmids

or cosmids. or cosmids.

Thegenetic The geneticconstruct constructcancan alsobe be also part part of of a genome a genome of a of a recombinant recombinant viral vector, viral vector,

including recombinant including recombinant adenovirus, adenovirus, recombinant recombinant adenovirus adenovirus associated associated virus andvirus and recombinant recombinant

vaccinia. The vaccinia. Thegenetic geneticconstruct construct cancan be part be part of the of the genetic genetic material material in attenuated in attenuated live live microorganisms microorganisms or or recombinant recombinant microbial microbial vectors vectors which which live in live in cells. cells. Thegenetic The geneticconstructs constructscancan comprise comprise regulatory regulatory elements elements for expression for gene gene expression of the of the codingsequences coding sequencesof of thethe nucleic nucleic acid. acid. The The regulatory regulatory elements elements can be can be a promoter, a promoter, an an enhancerananinitiation enhancer initiationcodon, codon,a stop a stopcodon, codon, orpolyadenylation or a a polyadenylation signal. signal.

-26-

Thenucleic The nucleicacid acidsequences sequencesmay may make make up a genetic up a genetic construct construct that that can be can be a vector. a vector.

Thevector The vectorcan canbebecapable capable of of expressing expressing an antigen an antigen in cell in the the cell of aof a mammal mammal in a quantity in a quantity

2023200517 01 Feb effective to effective to elicit elicitan an immune response immune response in in thethe mammal. mammal. The vector The vector can be can be recombinant. recombinant. The The vector can vector cancomprise comprise heterologous heterologous nucleic nucleic acid acid encoding encoding the antigen. the antigen. The can The vector vector be a can be a plasmid.The plasmid. Thevector vectorcancan be be useful useful for for transfecting transfecting cells cells with with nucleic nucleic acidacid encoding encoding an antigen, an antigen,

whichthe which thetransformed transformed host host cellcell is is cultured cultured andand maintained maintained underunder conditions conditions whereinwherein

expressionofofthe expression theantigen antigentakes takesplace. place. In some In embodiments, some embodiments, coding coding sequences sequences for a single for a single consensus consensus prostateprostate antigen antigen is is providedonona asingle provided singlevector. vector.In In some some embodiments, embodiments, coding coding sequences sequences for a for a multiple multiple consensusprostate consensus prostateantigen antigen areare provided provided on aon a single single vector. vector. In some In some embodiments, embodiments,

compositionsareareprovided compositions provided comprising comprising coding coding sequences sequences for a multiple for a multiple consensus consensus prostate prostate antigens ononmultiple antigens multiplevectors, vectors,either eitherone oneantigen antigen perper vector vector or multiple or multiple antigens antigens per vector. per vector.

In some In embodiments, some embodiments, coding coding sequences sequences for twofor or two more or more different different consensusconsensus prostate prostate antigens may antigens maybebeprovided provided on aon a single single vector. vector. In some In some embodiments, embodiments, thesequences the coding coding sequences mayhave may haveseparate separate promoters promoters controlling controlling expression. expression. In someInembodiments, some embodiments, the coding the coding sequencesmaymay sequences have have a single a single promoters promoters controlling controlling expression expression with anwith IRES an IRES sequence sequence separating coding separating codingsequence. sequence. The The presence presence of theofIRES the sequence IRES sequence results results in in the separate the separate

translation of translation of the the transcription product. InInsome transcription product. some embodiments, embodiments, the coding the coding sequences sequences may may haveaa single have singlepromoters promoterscontrolling controlling expression expression withwith coding coding sequence sequence encoding encoding a proteolytic a proteolytic

cleavagepeptide cleavage peptidesequence sequence separating separating coding coding sequences sequences of the of the antigens. antigens. A singleA translation single translation productisis produced product producedwhich which is then is then processed processed by protease by the the protease that recognizes that recognizes the protease the protease

cleavagesite cleavage site to to generate generateseparate separateprotein proteinmolecules. molecules. The The protease protease cleavecleave sites sites used used is is typically recognized typically recognizedbybya protease a protease endogenously endogenously present present in theincell the where cell where expression expression occurs. occurs.

In some In embodiments, some embodiments, a separate a separate coding coding sequence sequence for a protease for a protease may be included may be included to provideto provide for the for the production ofthe production of theprotease proteaseneeded needed to to process process the the polyprotein polyprotein translation translation product. product. In In someembodiment, some embodiment, vectors vectors comprise comprise coding coding sequences sequences for one, for two,one, two,four, three, three, four, five, sixfive, or six or all seven all consensusprostate seven consensus prostateantigens. antigens. In each In each and andevery everyinstance instance setset forthherein, forth herein,coding coding sequences sequences may may be be optimized optimized for for stability and stability and high levels of high levels of expression. expression.InInsome some instances, instances, codons codons are selected are selected to reduce to reduce

secondarystructure secondary structureformation formation of of thethe RNARNA such such as asformed that that formed due to due to intramolecular intramolecular bonding. bonding. Thevector The vectorcan cancomprise comprise heterologous heterologous nucleic nucleic acid encoding acid encoding an antigen an antigen and can and can further comprise further compriseananinitiation initiationcodon, codon,which which can can be upstream be upstream of theof the antigen antigen codingcoding sequence, sequence,

and aa stop and stop codon, codon,which whichcancan be downstream be downstream of the of the antigen antigen coding coding sequence. sequence. The initiation The initiation

and termination and terminationcodon codon cancan be frame be in in frame with with the antigen the antigen codingcoding sequence. sequence. Thecanvector The vector also can also

-27-

Feb 2023 comprisea apromoter comprise promoter that that is is operably operably linked linked to the to the antigen antigen coding coding sequence. sequence. The promoter The promoter

linkedtotothe operablylinked operably theantigen antigencoding coding sequence sequence cana be can be a promoter promoter from simian simian40virus from virus 40 (SV40), aa mouse (SV40), mammary mouse mammary tumor tumor virus virus (MMTV) (MMTV) promoter, promoter, a human a human immunodeficiency immunodeficiency

virus (HIV) virus (HIV)promoter promoter such such as the as the bovine bovine immunodeficiency immunodeficiency virus virus (BIV) (BIV) long longrepeat terminal terminal repeat 2023200517 01 (LTR)promoter, (LTR) promoter, a Moloney a Moloney virusvirus promoter, promoter, an leukosis an avian avian leukosis viruspromoter, virus (ALV) (ALV) promoter, a a cytomegalovirus (CMV) cytomegalovirus (CMV)promoter promoter suchasasthe such theCMV CMV immediate immediate early early promoter, promoter, Epstein Epstein Barr Barr

virus (EBV) virus (EBV)promoter, promoter, orRous or a a Rous sarcoma sarcoma virus virus (RSV) (RSV) promoter. promoter. Thecan The promoter promoter also be can a also be a promoter from promoter from aa human humangene genesuch suchasas human humanactin, actin, human humanmyosin, myosin,human human hemoglobin, hemoglobin,

humanmuscle human muscle creatine, creatine, or human or human metalothionein. metalothionein. The promoter The promoter cana tissue can also be also bespecific a tissue specific promoter,such promoter, suchasasa amuscle muscle or or skin skin specific specific promoter, promoter, natural natural or synthetic. or synthetic. Examples Examples of suchof such promotersarearedescribed promoters described in in US US patent patent application application publication publication no.US20040175727, no. US20040175727, the the contents of contents ofwhich whichareareincorporated incorporated herein herein in its in its entirety. entirety.

Thevector The vectorcan canalso alsocomprise comprise a polyadenylation a polyadenylation signal, signal, whichwhich can be can be downstream downstream of of the consensus the consensusprostate prostateantigen antigen coding coding sequence. sequence. The polyadenylation The polyadenylation signal signal can be a can SV40be a SV40 polyadenylation signal, polyadenylation signal,LTR LTR polyadenylation polyadenylation signal, signal,bovine bovinegrowth growthhormone hormone (bGH) (bGH)

polyadenylation signal, polyadenylation signal,human human growth growth hormone (hGH)polyadenylation hormone (hGH) polyadenylationsignal, signal, or or humanp human ß-

globin polyadenylation globin polyadenylation signal.TheThe signal. SV40SV40 polyadenylation polyadenylation signal signal can be acan be a polyadenylation polyadenylation

signal from signal froma apCEP4 pCEP4 vector vector (Invitrogen, (Invitrogen, San Diego, San Diego, CA). CA). Thevector The vectorcan canalso alsocomprise comprise an enhancer an enhancer upstream upstream of the of the consensus consensus prostateprostate antigen antigen coding sequence. coding sequence. The The enhancer enhancercan can be be necessary necessary for for DNA expression. The DNA expression. Theenhancer enhancercan canbebe humanactin, human actin, human myosin,human human myosin, humanhemoglobin, hemoglobin, human human muscle muscle creatine creatine or or a viral enhancer a viral enhancer such as such as one one from from CMV, HA,RSV CMV, HA, RSV or or EBV. EBV. Polynucleotide Polynucleotide function function enhances enhances are are described described

in U.S. in Patent Nos. U.S. Patent Nos.5,593,972, 5,593,972,5,962,428, 5,962,428, and and W094/016737, WO94/016737, the contents the contents of each of each are fully are fully incorporatedbybyreference. incorporated reference. Thevector The vectorcan canalso alsocomprise comprise a mammalian a mammalian origin origin of replication of replication in to in order order to maintain maintain

the vector the extrachromosomally vector extrachromosomally and produce and produce multiple multiple copies copies of the in of the vector vector in aThe a cell. cell. The vector can vector can be be pVAXI, pCEP4 pVAX1, pCEP4 or or pREP4 pREP4 from from Invitrogen Invitrogen (San (San Diego, Diego, CA), CA), which which cancan

comprisethe comprise theEpstein Epstein Barr Barr virus virus origin origin of replication of replication andand nuclear nuclear antigen antigen EBNA-1 EBNA-1 coding coding region, which region, whichcan canproduce produce highhigh copycopy episomal episomal replication replication without without integration. integration. The The backbone backbone of the of the vector can be vector can bepAV0242. pAV0242. The vector The vector can becan be a replication a replication defective defective adenovirus adenovirus type 5 type 5 (Ad5)vector. (Ad5) vector. Thevector The vectorcan canalso alsocomprise comprise a regulatory a regulatory sequence, sequence, whichwhich can be can wellbe well for suited suited for geneexpression gene expressioninina amammalian mammalian or human or human cellwhich cell into into the which the is vector vector is administered. administered. The The

-28- consensusprostate consensus prostateantigen antigen coding coding sequence sequence can comprise can comprise a codon, a codon, which which can allowcan moreallow more efficient transcription efficient transcription of of the the coding sequence coding sequence in in thehost the host cell. cell.

Thevector The vectorcan canbebepSE420 pSE420 (Invitrogen, (Invitrogen, San Diego, San Diego, Calif.), Calif.), which which can be can used be forused for protein production protein productionininEscherichia Escherichia coli coli (E.(E. coli).TheThe coli). vector vector can can also also be pYES2 be pYES2 (Invitrogen, (Invitrogen,

San Diego, San Diego,Calif.), Calif.), which whichcancan be be used used for for protein protein production production in Saccharomyces in Saccharomyces cerevisiae cerevisiae

strains ofofyeast. strains The yeast. vector The cancan vector alsoalso be of be the of MAXBACTM the completecomplete baculovirus baculovirus expression expression

system(Invitrogen, system (Invitrogen,SanSan Diego, Diego, Calif.), Calif.), which which can can be used be used for protein for protein production production in insect in insect

cells. The cells. vector can The vector canalso alsobebepcDNA pcDNAI or IpcDNA3 or pcDNA3 (Invitrogen, (Invitrogen, SanCalif.), San Diego, Diego, which Calif.), which maybebeused may usedforforprotein protein production production in mammalian in mammalian cellsassuch cells such as Chinese Chinese hamster hamster ovary ovary (CHO)cells. (CHO) cells.TheThe vector vector can can be expression be expression vectors vectors or systems or systems to produce to produce protein protein by by routine routine techniquesand techniques andreadily readilyavailable availablestarting startingmaterials materials including including Sambrook Sambrook et al.,etMolecular al., Molecular Cloning and Cloning and Laboratory Laboratory Manual, Manual, Second SecondEd. , Cold Ed.Cold Spring Spring Harbor Harbor (1989),which (1989) ,which is is

incorporatedfully incorporated fullybybyreference. reference. Vaccines may Vaccines may comprise comprise onemore one or or more of the of the prostate prostate antigens antigens set herein set forth forth herein and/or and/or

vaccinesmay vaccines may comprise comprise one one or more or more nucleic nucleic acid sequences acid sequences that one that encode encode oneoforthe or more more of the consensusprostate consensus prostateantigen antigen selected selected from from thisthis group. group. Vaccines Vaccines may comprise may comprise oneofor one or more more of the consensus the consensusprostate prostateantigens antigens setset forthherein forth herein in in combination combination with with otherother immunogenic immunogenic

prostate proteins prostate proteins with withsequences sequences other other than than the the consensus consensus sequences sequences disclosed disclosed herein herein including native including nativesequences sequences and/or and/or vaccines vaccines may comprise may comprise one or one more or more acid nucleic nucleic acid sequences sequences

that encode that oneorormore encode one more of of thethe consensus consensus prostate prostate antigens antigens selected selected fromgroup from this this group in in combinationwith combination with nucleic nucleic acidacid molecules molecules that that encode encode other other prostate prostate antigens antigens with sequences with sequences

other than other than the the consensus consensussequences sequences disclosed disclosed herein. herein.

Whilenot While notbeing beingbound bound by scientific by scientific theory, theory, a vaccine a vaccine that that can can be used be used to elicit to elicit an an immune immune response response (humoral, (humoral, cellular, cellular, or both) or both) broadly broadly against against prostate prostate cancercancer cells cells may may compriseone comprise oneor ormore more of the of the following following nucleic nucleic acid acid sequences sequences that encodes that encodes one one or more or more proteins selected proteins selected from fromthethegroup group consisting consisting of: of: consensus, consensus, PSA antigen PSA antigen 1, consensus, 1, consensus, PSA PSA antigen 2,2,consensus, antigen consensus,PSMA antigen 1, PSMA antigen 1, consensus, consensus, PSMA antigen 2, PSMA antigen 2, consensus consensus STEAP STEAP antigen 1,1,consensus antigen consensus STEAP antigen 22 and STEAP antigen and consensus consensus PSCA PSCAantigen antigen1.1. Coding Codingsequences sequences mayalso may alsoinclude includethose those provided provided herein herein thatthat comprise comprise homologous homologous sequences, sequences, fragments,fragments, and and homologoussequences homologous sequencesofoffragments. fragments. Someembodiments Some embodiments provide provide methods methods of of generatingimmune generating immune responses responses against against prostate prostate

cancer cells cancer cells comprise compriseadministering administering to an to an individual individual one one or more or more compositions compositions which which collectively comprise collectively compriseoneone or or more more coding coding sequences sequences or combinations or combinations described described herein. herein. Some Some embodiments embodiments provide provide methods methods of prophylactically of prophylactically vaccinating vaccinating an individual an individual against against prostate prostate

-29- cancer comprise cancer compriseadministering administering one one or more or more compositions compositions which collectively which collectively comprise comprise one or one or more coding more coding sequences sequences or or combinations described herein. combinations described herein. Some Some embodiments provide embodiments provide

2023200517 01 Feb methodsofoftherapeutically methods therapeutically vaccinating vaccinating an individual an individual has prostate has prostate cancer cancer that comprise that comprise

administeringone administering oneor ormore more compositions compositions which which collectively collectively comprise comprise one coding one or more or more coding sequencesororcombinations sequences combinations described described herein. herein.

4. Pharmaceutical 4. Pharmaceutical compositions compositions

Providedherein Provided hereinarearepharmaceutical pharmaceutical compositions compositions according according to the present to the present invention invention

which comprise which comprise about about 11 nanogram nanogramtoto about about 10 10 mg mgofofDNA. DNA.In In some some embodiments, embodiments,

pharmaceuticalcompositions pharmaceutical compositions according according to thetopresent the present invention invention comprise comprise from between: from between: 1) at 1) at least 10, least 10, 15, 15, 20, 20, 25, 25, 30, 30, 35, 35, 40, 40, 45, 45, 50, 50, 55, 60, 65, 55, 60, 65, 70, 70, 75, 75, 80, 80, 85, 85, 90, 90, 95 95 or or 100 100nanograms, nanograms,or or atleast at least 1, 1,5,5,10, 10,15,20,25,30, 15, 20, 25, 30,35,40,45,50, 35, 40, 45, 50, 55, 55, 60,65,70,75, 60, 65, 70, 75, 80, 80, 85, 85, 90,95,100, 105, 110, 90, 95,100, 105, 110, 115,120,125,130,135,140,145,150,155,160,165,170,175,180,185,190,195,200, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, 195, 200,

205,210,215,220,225,230,235,240,245,250,255,260,265,270,275,280,285,290, 205, 210, 215, 220, 225, 230, 235, 240, 245, 250, 255, 260, 265, 270, 275, 280, 285, 290,

295,300,305,310,315,320,325,330,335,340,345,350,355,360,365,370,375,380, 295, 300, 305, 310, 315, 320, 325, 330, 335, 340, 345, 350, 355, 360, 365, 370, 375, 380,

385,390,395,400,405,410,415,420,425,430,435,440,445,450,455,460,465,470, 385, 390, 395, 400, 405, 410, 415, 420, 425, 430, 435, 440, 445, 450, 455, 460, 465, 470,

475,480,485,490,495,500,605,610,615,620,625,630,635,640,645,650,655,660, 475, 480, 485, 490, 495, 500, 605, 610, 615, 620, 625, 630, 635, 640, 645, 650, 655, 660,

665,670,675,680,685,690,695,700,705,710,715,720,725,730,735,740,745,750, 665, 670, 675, 680, 685, 690, 695, 700, 705, 710, 715, 720, 725, 730, 735, 740, 745, 750,

755,760,765,770,775,780,785,790,795,800,805,810,815,820,825,830,835,840, 755, 760, 765, 770, 775, 780, 785, 790, 795, 800, 805, 810, 815, 820, 825, 830, 835, 840,

845,850,855,860,865,870,875,880,885,890,895.900,905,910,915,920,925,930, 845, 850, 855, 860, 865, 870, 875, 880, 885, 890, 895. 900, 905, 910, 915, 920, 925, 930,

935, 940, 935, 940,945, 945,950, 950,955, 955,960, 960,965, 965, 970, 970, 975,975, 980,980, 985,985, 990, 990, 9951000 995 or or micrograms, 1000 micrograms, or at or at least 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5 or 10 mg or more; and 2) up least 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5 or 10 mg or more; and 2) up

to and to including15, and including 15,20, 20,25, 25,30, 30,35, 35,40, 40,45, 45,50, 50,55, 55,60, 60,65,65,70,70,75, 75,80,80,85,85,90,90,9595or or100100 nanograms,or orupup nanograms, to to andand including including 1, 10, 1, 5, 5, 10, 15, 15, 20, 20, 25, 25, 30, 30, 35, 35, 40, 40, 45, 45, 50, 50, 55, 55, 60, 60, 65, 65, 70, 70, 75, 75, 80, 85, 80, 85, 90, 90, 95, 95,100, 105, 110, 100, 105, 110, 115, 115,120, 120,125, 125,130, 130,135, 135, 140, 140, 145, 145, 150,150, 155,155, 160, 160, 165, 165, 170, 170, 175, 175, 180,185,190,195,200,205,210,215,220,225,230,235,240,245,250,255,260,265, 180, 185, 190, 195, 200, 205, 210, 215, 220, 225, 230, 235, 240, 245, 250, 255, 260, 265,

270,275,280,285,290,295,300,305,310,315,320,325,330,335,340,345,350,355, 270, 275, 280, 285, 290, 295, 300, 305, 310, 315, 320, 325, 330, 335, 340, 345, 350, 355,

360,365,370,375,380,385,390,395,400,405,410,415,420,425,430,435,440,445, 360, 365, 370, 375, 380, 385, 390, 395, 400, 405, 410, 415, 420, 425, 430, 435, 440, 445,

450,455,460,465,470,475,480,485,490,495,500,605,610,615,620,625,630,635, 450, 455, 460, 465, 470, 475, 480, 485, 490, 495, 500, 605, 610, 615, 620, 625, 630, 635,

640,645,650,655,660,665,670,675,680,685,690,695,700,705,710,715,720,725, 640, 645, 650, 655, 660, 665, 670, 675, 680, 685, 690, 695, 700, 705, 710, 715, 720, 725,

730,735,740,745,750,755,760,765,770,775,780,785,790,795,800,805,810,815, 730, 735, 740, 745, 750, 755, 760, 765, 770, 775, 780, 785, 790, 795, 800, 805, 810, 815,

820,825,830,835,840,845,850,855,860,865,870,875,880,885,890,895.900,905, 820, 825, 830, 835, 840, 845, 850, 855, 860, 865, 870, 875, 880, 885, 890, 895. 900, 905,

910,915,920,925,930,935,940,945,950,955,960,965,970,975,980,985,990,995,or 910, 915, 920, 925, 930, 935, 940, 945, 950, 955, 960, 965, 970, 975, 980, 985, 990, 995, or

1000micrograms, 1000 micrograms, or up or up to and to and including including 1.5, 1.5, 2, 2.5, 2, 2.5, 3, 3.5, 3, 3.5, 4, 4, 4.5,5, 5,5.5, 4.5, 5.5,6,6,6.5, 6.5,7,7, 7.5, 7.5, 8, 8, 8.5, 8.5,

-30-

9, 9.5 9, 9.5 or 10 mg. or 10 mg.InInsome some embodiments, embodiments, pharmaceutical pharmaceutical compositions compositions according according to the to the present invention present inventioncomprise comprise about about5 5nanogram nanogram to to about about 10 10mg mg of ofDNA. In some DNA. In some

2023200517 01 Feb embodiments, embodiments, pharmaceutical pharmaceutical compositions compositions according according to the invention to the present present invention comprise comprise about 25 about 25 nanogram to about nanogram to about 55 mg of DNA. mg of Insome DNA. In someembodiments, embodiments,thethepharmaceutical pharmaceutical compositions contain compositions contain about about 50 50 nanograms to about nanograms to 1 mg about 1 of DNA. mg of DNA. InInsome someembodiments, embodiments, thethe

pharmaceutical compositions pharmaceutical compositions contain contain about about 0.1 0.1totoabout about500 500micrograms micrograms of ofDNA. In some DNA. In some

embodiments, embodiments, thethe pharmaceutical pharmaceutical compositions compositions containcontain about 1 about 1 to350about to about 350 micrograms micrograms of of DNA.InInsome DNA. someembodiments, embodiments,thethe pharmaceuticalcompositions pharmaceutical compositionscontain containabout about5 5toto about about 250 250 microgramsofofDNA. micrograms DNA.InInsome someembodiments, embodiments, thethe pharmaceutical pharmaceutical compositions compositions containabout contain about 10 to 10 to about about 200 200 micrograms micrograms of of DNA. In some DNA. In someembodiments, embodiments,the thepharmaceutical pharmaceutical compositions contain compositions contain about about 15 15 to toabout about150 150micrograms micrograms of of DNA. In some DNA. In some embodiments, embodiments,the the pharmaceutical compositions pharmaceutical compositions contain contain about about 20 20 to toabout about100 100micrograms micrograms of of DNA. In some DNA. In some

embodiments, embodiments, thethe pharmaceutical pharmaceutical compositions compositions containcontain about 25about 25 to to about 75 about 75 micrograms micrograms of of DNA.InInsome DNA. someembodiments, embodiments,thethe pharmaceuticalcompositions pharmaceutical compositionscontain containabout about3030toto about about 50 50 microgramsofofDNA. micrograms DNA.InInsome someembodiments, embodiments, thethe pharmaceutical pharmaceutical compositions compositions containabout contain about 35 to 35 to about about 40 40 micrograms of DNA. micrograms of DNA. InInsome someembodiments, embodiments, thethe pharmaceuticalcompositions pharmaceutical compositions contain about contain about 100 100 to toabout about200 200microgram microgram DNA. DNA. InInsome someembodiments, embodiments, thethe pharmaceutical pharmaceutical

compositions comprise compositions comprise about about 10 10 microgram microgramtoto about about 100 100 micrograms microgramsofofDNA. DNA.In In some some

embodiments, the embodiments, the pharmaceutical pharmaceutical compositions compositions comprise comprise about about 20 20 micrograms microgramstotoabout about 80 80 microgramsofofDNA. micrograms DNA.In In some some embodiments, embodiments, the the pharmaceutical pharmaceutical compositions compositions comprise comprise

about 25 about 25 micrograms to about micrograms to about 60 60 micrograms of DNA. micrograms of DNA.InInsome some embodiments, embodiments, thethe

pharmaceutical compositions pharmaceutical compositions comprise comprise about about 30 30 nanograms nanogramstoto about about 50 50 micrograms microgramsofofDNA. DNA. In some In embodiments,the some embodiments, the pharmaceutical pharmaceutical compositions compositions comprise compriseabout about 35 35 nanograms nanogramstoto about 45 about 45 micrograms of DNA. micrograms of DNA.In Insome some preferredembodiments, preferred embodiments, thepharmaceutical the pharmaceutical compositionscontain compositions contain about about 0.1 0.1 to about to about 500 500 micrograms micrograms of DNA.of InDNA. In some some preferred preferred embodiments, embodiments, thethe pharmaceutical pharmaceutical compositions compositions containcontain about 1 about 1 to350about to about 350 micrograms micrograms of of DNA.InInsome DNA. somepreferred preferred embodiments, embodiments,the thepharmaceutical pharmaceutical compositions compositionscontain contain about about 25 25 to to about 250 about microgramsofofDNA. 250 micrograms DNA.InInsome somepreferred preferredembodiments, embodiments,the thepharmaceutical pharmaceutical compositions contain compositions contain about about 100 100 to toabout about200 200microgram microgram DNA. DNA.

Thepharmaceutical The pharmaceutical compositions compositions according according to the to the present present invention invention are formulated are formulated

accordingtotothe according themode modeof of administration administration to used. to be be used. In cases In cases wherewhere pharmaceutical pharmaceutical

compositionsareareinjectable compositions injectablepharmaceutical pharmaceutical compositions, compositions, they they are are sterile, sterile, pyrogen pyrogen free free and and particulate free. particulate free. An isotonic formulation An isotonic formulationis ispreferably preferably used. used. Generally, Generally, additives additives for for isotonicity can isotonicity can include includesodium sodium chloride, chloride, dextrose, dextrose, mannitol, mannitol, sorbitol sorbitol and lactose. and lactose. In some In some

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Feb 2023 cases, isotonic cases, isotonic solutions solutions such suchasasphosphate phosphate buffered buffered saline saline are are preferred. preferred. Stabilizers Stabilizers include include

gelatin and gelatin and albumin. albumin.InInsome some embodiments, embodiments, a vasoconstriction a vasoconstriction agent agent is addedisto added the to the formulation. formulation.

Preferablythe Preferably thepharmaceutical pharmaceutical composition composition is a vaccine, is a vaccine, and preferably and more more preferably a DNA a DNA 2023200517 01 vaccine. vaccine.

The vaccine The vaccine may maybe be aa DNA DNAvaccine. vaccine.The The DNA DNA vaccine vaccine may may comprise comprise a plurality a plurality of of the same the sameorordifferent differentplasmids plasmidscomprising comprising nucleic nucleic acid acid coding coding sequences sequences for one for one or or more of more of consensus prostate consensus prostate antigens. antigens.The TheDNA vaccine may DNA vaccine maycomprise compriseone oneorormore morenucleic nucleic acid acid sequences that sequences that encode encode one one or ormore more of of consensus consensus prostate prostateantigens. When antigens. When the theDNA vaccine DNA vaccine

comprisescoding comprises coding sequences sequences of more of more thanconsensus than one one consensus prostateprostate antigensantigens all such all such sequences sequences

maybebepresent may presentonon a single a single plasmid, plasmid, or each or each suchsuch sequences sequences may bemay be present present on a different on a different

plasmids. plasmids.

In some In embodiments,vaccines some embodiments, vaccines may maycomprise comprisenucleic nucleicacid acid sequences sequences that that encode one encode one

or more or ofconsensus more of consensus prostate prostate antigens antigens in combination in combination withorone with one moreorofmore of consensus consensus

prostate antigens. prostate antigens. DNA DNA vaccines vaccines are are disclosed disclosed inPatent in US US Patent Nos. 5,593,972, Nos. 5,593,972, 5,739,118, 5,739,118, 5,817,637, 5,817,637,

5,830,876, 5,962,428, 5,830,876, 5,962,428,5,981,505, 5,981,505, 5,580,859, 5,580,859, 5,703,055, 5,703,055, and 5,676,594, and 5,676,594, which which are are incorporatedherein incorporated hereinfully fullybybyreference. reference.TheThe DNA DNA vaccine vaccine can further can further comprisecomprise elements elements or or reagents that reagents that inhibit inhibit it it from integrating into from integrating into the the chromosome. chromosome. The vaccine The vaccine can be can be an an RNA of RNA of the prostate the antigen. The prostate antigen. TheRNARNA vaccine vaccine can becan be introduced introduced into into the the cell. cell. Thevaccine The vaccinecancan be be a recombinant a recombinant vaccine vaccine comprising comprising the genetic the genetic construct construct or or antigen antigen describedabove. described above.TheThe vaccine vaccine can also can also comprise comprise one or one moreorconsensus more consensus prostate in prostate antigens antigens in the form the formofofone oneorormore more protein protein subunits, subunits, or one or one or more or more attenuated attenuated viral viral particles particles comprising comprising

one or one or more moreconsensus consensus prostate prostate antigens. antigens. The attenuated The attenuated vaccine vaccine can be can be attenuated attenuated live live vaccines, killed vaccines, killed vaccines vaccinesand andvaccines vaccines that that useuse recombinant recombinant vectors vectors to deliver to deliver foreign foreign genes genes that encode that oneorormore encode one more consensus consensus prostate prostate antigens, antigens, and as and well well as subunit subunit and glycoprotein and glycoprotein

vaccines. Examples vaccines. Examplesof of attenuated attenuated livelive vaccines, vaccines, those those usingusing recombinant recombinant vectorsvectors to deliver to deliver

prostate antigens, prostate antigens, subunit subunitvaccines vaccinesandand glycoprotein glycoprotein vaccines vaccines are described are described in Patent in U.S. U.S. Patent Nos.: 4,510,245;4,797,368; Nos.: 4,510,245; 4,797,368; 4,722,848; 4,722,848; 4,790,987; 4,790,987; 4,920,209; 4,920,209; 5,017,487; 5,017,487; 5,077,044; 5,077,044;

5,110,587;5,112,749; 5,110,587; 5,112,749;5,174,993; 5,174,993; 5,223,424; 5,223,424; 5,225,336; 5,225,336; 5,240,703; 5,240,703; 5,242,829; 5,242,829; 5,294,441; 5,294,441;

5,294,548;5,310,668; 5,294,548; 5,310,668;5,387,744; 5,387,744; 5,389,368; 5,389,368; 5,424,065; 5,424,065; 5,451,499; 5,451,499; 5,453,3 5,453,3 64; 5,462,734; 64; 5,462,734;

5,470,734;5,474,935; 5,470,734; 5,474,935;5,482,713; 5,482,713; 5,591,439; 5,591,439; 5,643,579; 5,643,579; 5,650,309; 5,650,309; 5,698,202; 5,698,202; 5,955,088; 5,955,088;

6,034,298;6,042,836; 6,034,298; 6,042,836;6,156,319 6,156,319 and and 6,589,529, 6,589,529, which which areincorporated are each each incorporated herein byherein by reference. reference.

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Feb 2023 vaccineprovided Thevaccine The provided maymay be used be used immuneimmune to induce to induce responsesresponses including including therapeutic therapeutic

or prophylactic or immune prophylactic immune responses. responses. Antibodies Antibodies and/or and/or killer killer T cellsT may cellsbe may be generated generated which which are directed are directed to to the the consensus consensusprostate prostateantigen. antigen.SuchSuch antibodies antibodies and cells and cells may may be be isolated. isolated.

Thevaccine The vaccinecancan furthercomprise further comprise a pharmaceutically a pharmaceutically acceptable acceptable excipient. excipient. The The 2023200517 01 pharmaceuticallyacceptable pharmaceutically acceptable excipient excipient can can be functional be functional molecules molecules as vehicles, as vehicles, adjuvants, adjuvants,

carriers, or carriers, or diluents. diluents. The pharmaceutically The pharmaceutically acceptable acceptable excipient excipient cana be can be a transfection transfection

facilitating agent, facilitating agent, which caninclude which can includesurface surfaceactive activeagents, agents, such such as immune-stimulating as immune-stimulating

complexes (ISCOMS), complexes (ISCOMS),Freunds Freunds incomplete incomplete adjuvant,LPS adjuvant, LPS analog analog includingmonophosphoryl including monophosphoryl lipid A, lipid A, muramyl peptides, muramyl peptides, quinone quinone analogs, analogs, vesicles vesicles such such as squalene as squalene and squalene, and squalene,

hyaluronicacid, hyaluronic acid,lipids, lipids, liposomes, liposomes,calcium calcium ions, ions, viralproteins, viral proteins,polyanions, polyanions, polycations, polycations, or or nanoparticles, ororother nanoparticles, otherknown known transfection transfection facilitating facilitating agents. agents.

Thetransfection The transfectionfacilitating facilitating agent agentisis aa polyanion, polyanion,polycation, polycation,including including poly-L poly-L-

glutamate(LGS), glutamate (LGS),or or lipid.TheThe lipid. transfection transfection facilitating facilitating agent agent is poly-L-glutamate, is poly-L-glutamate, and and more more preferably, the preferably, the poly-L-glutamate poly-L-glutamate is present is present in in thethe vaccine vaccine at aatconcentration a concentration less less thanthan 6 6 mg/ml.TheThe mg/ml. transfection transfection facilitating facilitating agent agent can can alsoalso include include surface surface active active agents agents such such as as immune-stimulating complexes immune-stimulating complexes(ISCOMS), (ISCOMS), Freunds Freunds incomplete incomplete adjuvant, adjuvant, LPSLPS analog analog

including monophosphoryl including monophosphoryllipidlipid A, muramyl A, muramyl peptides, peptides, quinone quinone analogs analogs and andsuch vesicles vesicles as such as squaleneand squalene andsqualene, squalene, andand hyaluronic hyaluronic acid acid can also can also be used be used administered administered in conjunction in conjunction with with the genetic the construct. InIn some genetic construct. someembodiments, embodiments, the vector the DNA DNA vaccines vector vaccines can also can alsoa include include a transfection facilitating transfection facilitating agent suchasaslipids, agent such lipids, liposomes, liposomes,including includinglecithin lecithinliposomes liposomes or other or other

liposomes known liposomes knowninin the the art, art,asasa DNA-liposome a DNA-liposome mixture mixture (see (see for forexample example W09324640), W09324640),

calciumions, calcium ions,viral viral proteins, proteins, polyanions, polyanions,polycations, polycations,or or nanoparticles, nanoparticles, or other or other known known

transfection facilitating transfection facilitating agents. agents. Preferably, the transfection Preferably, the transfection facilitating facilitating agent agent isis aa polyanion, polyanion,

polycation, including polycation, includingpoly-L-glutamate poly-L-glutamate (LGS), (LGS), or lipid. or lipid. Concentration Concentration of the transfection of the transfection

agent in agent in the the vaccine vaccineisis less less than than 44 mg/ml, mg/ml,less lessthan than2 mg/ml, 2 mg/ml, lessless than than 1 mg/ml, 1 mg/ml, less less than than 0.750 mg/ml, 0.750 mg/ml,less lessthan than 0.500 0.500 mg/ml, mg/ml, less less thanthan 0.2500.250 mg/ml, mg/ml, less0.100 less than than mg/ml, 0.100 less mg/ml, thanless than 0.050 mg/ml, 0.050 mg/ml,or orless lessthan than0.010 0.010 mg/ml. mg/ml.

Thepharmaceutically The pharmaceutically acceptable acceptable excipient excipient may may be an be an adjuvant. adjuvant. The adjuvant The adjuvant may be may be other genes other genesthat that are are expressed expressedininalternative alternativeplasmid plasmid or or areare delivered delivered as proteins as proteins in in combinationwith combination with thethe plasmid plasmid above above in vaccine. in the the vaccine. The adjuvant The adjuvant may be from may be selected selected the from the groupconsisting group consistingof: of:-interferon(IFN- a-interferon(IFN- a),3-interferon o ß-interferon (IFN-P), (IFN-), y-interferon, y-interferon, plateletplatelet derived derived

growth factor growth factor (PDGF), TNFa,TNFß, (PDGF), TNF, TNFP, GM-CSF, GM-CSF, epidermal epidermal growth growth factor factor (EGF), (EGF), cutaneous cutaneous T T cell-attracting chemokine cell-attracting chemokine(CTACK), epithelial thymus-expressed (CTACK), epithelial thymus-expressed chemokine chemokine (TECK), (TECK),

mucosae-associated epithelial mucosae-associated epithelial chemokine chemokine (MEC), IL-12, IL-15, (MEC), IL-12, IL-15, MHC, CD80,CD86 MHC, CD80,CD86 including including

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Feb 2023 IL-15 having IL-15 havingthethesignal sequence signalsequence deleted deleted and and optionally optionally including including the signal the signal peptide peptide from from IgE. The IgE. The adjuvant maybe adjuvant may be IL-12, IL-12, IL-15, IL-15, IL-28, IL-28, CTACK, TECK, CTACK, TECK, plateletderived platelet derivedgrowth growth factor (PDGF), factor (PDGF), TNFc, TNF, TNF, TNFß, GM-CSF, GM-CSF, epidermal epidermal growth growth factor factor (EGF), (EGF), IL-1,IL-1, IL-2, IL-2, IL-4, IL-4, IL-IL

5, IL-6, 5, IL-6, IL-10, IL-12, IL-18, IL-10, IL-12, IL-18,orora acombination combination thereof. thereof.

2023200517 01 Othergenes Other geneswhich which maymay be useful be useful adjuvants adjuvants include include those encoding: those encoding: MCP-1, MCP-1, MIP-la, MIP-la, MIP-ip, IL-8, MIP-1p, IL-8, RANTES, L-selectin, P-selectin, RANTES, L-selectin, P-selectin, E-selectin, E-selectin,CD34, CD34,GlyCAM-1, MadCAM-1, GlyCAM-1, MadCAM-1,

LFA-1, VLA-1, LFA-1, VLA-1,Mac-1, Mac-1,pl50.95, p150.95,PECAM, PECAM, ICAM-1, ICAM-1, ICAM-2, ICAM-2, ICAM-3, ICAM-3, CD2,M-LFA-3, CD2, LFA-3, M CSF,G-CSF, CSF, G-CSF, IL-4, IL-4, mutant mutant formsforms of IL-18, of IL-18, CD40, CD40, CD40L,growth CD40L, vascular vascular growth factor, factor, fibroblast fibroblast growthfactor, growth factor, IL-7, IL-7,nerve nervegrowth growth factor, factor, vascular vascular endothelial endothelial growth growth factor, factor, Fas, Fas, TNF TNF receptor, Flt, receptor, Flt,Apo-1, p55, Apo-1, WSL-1, p55, WSL-1,DR3, DR3, TRAMP, Apo-3,AIR, TRAMP, Apo-3, AIR,LARD, LARD, NGRF, NGRF, DR4, DR4, DR5, DR5, KILLER,TRAIL-R2, KILLER, TRAIL-R2, TRICK2, TRICK2, DR6, DR6, Caspase Caspase ICE, c-jun, ICE, Fos, Fos, c-jun, Sp-1,Sp-1, Ap-1,Ap-1, Ap-2, Ap-2, p38,p38,

p65Rel, MyD88, p65Rel, MyD88,IRAK, IRAK, TRAF6, TRAF6, IkB,IkB, Inactive Inactive NIK, NIK, SAP SAP K, SAP-1, K, SAP-1, JNK, JNK, interferon interferon

response genes, response genes, NFkB, Bax, TRAIL, NFkB, Bax, TRAIL,TRAILrec, TRAILrec, TRAILrecDRC5, TRAILrecDRC5, TRAIL-R3, TRAIL-R3, TRAIL-R4, TRAIL-R4,

RANK, RANK RANK, RANKLIGAND, LIGAND,Ox40, Ox40, Ox40 Ox40 LIGAND, LIGAND, NKG2D, NKG2D, MICA, MICA, MICB, MICB, NKG2A, NKG2A, NKG2B, NKG2C, NKG2B, NKG2C, NKG2E, NKG2E, NKG2F,NKG2F, TAPI, TAP1, TAP2 and TAP2 and functional functional fragmentsfragments thereof. thereof.

Thevaccine The vaccinecancan further further comprise comprise a genetic a genetic vaccine vaccine facilitator facilitator agent agent as described as described in in U.S. Serial U.S. Serial No. No.021,579 021,579 filedApril filed April 1, 1, 1994, 1994, which which is fully is fully incorporated incorporated by reference. by reference.

5. Methods 5. MethodsofofDelivery Delivery Providedherein Provided hereinisisa amethod methodforfor delivering delivering the the pharmaceutical pharmaceutical formulations, formulations,

preferably vaccines, preferably vaccines,for forproviding providinggenetic genetic constructs constructs and and consensus consensus prostate prostate antigen antigen which which compriseepitopes comprise epitopesthat thatmake make themthem particular particular effective effective immunogens immunogens against against which an which immune an immune responsetotoprostate response prostatecancer cancercells cellscan canbebeinduced. induced. The The method method of delivering of delivering the vaccine, the vaccine, or or vaccination, can vaccination, canbebeprovided providedto to induce induce a therapeutic a therapeutic and/or and/or prophylactic prophylactic immuneimmune response. response.

Thevaccine The vaccinecancan be be delivered delivered to an to an individual individual to modulate to modulate the activity the activity ofmammal's of the the mammal's immunesystem immune systemand andenhance enhancethe theimmune immune response. response.

Upon delivery Upon delivery of of thethe vaccine vaccine to the to the mammal, mammal, and thereupon and thereupon the into the vector vector theinto the cells cells

of the of the mammal, mammal, thethe transfected transfected cells cells will will express express and and secrete secrete the corresponding the corresponding prostate prostate

consensusprotein. consensus protein.These These secreted secreted proteins, proteins, or synthetic or synthetic antigens, antigens, will will be recognized be recognized by theby the immunesystem, immune system,which whichwill will mount mountananimmune immune response response thatcan that caninclude: include: antibodies antibodies made made

against the against the antigens, antigens, and andT-cell T-cellresponse responsespecifically specifically against against thethe antigen. antigen. In some In some examples, examples,

a mammal a mammal vaccinated vaccinated with with the vaccines the vaccines discussed discussed herein herein willa have will have primeda immune primedsystem. immune system. Thevaccine The vaccinecancan be be delivered delivered to an to an individual individual to modulate to modulate the activity the activity ofindividual's of the the individual's immunesystem immune systemthereby therebyenhancing enhancingthe theimmune immune response. response.

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Feb 2023 Thevaccine The vaccinecancan be be delivered delivered in the in the form form of aof a DNA DNA vaccine vaccine and methods and methods of delivering of delivering

a DNA a DNA vaccines vaccines are are described described in U.S. in U.S. Patent Patent Nos. Nos. 4,945,050 4,945,050 and 5,036,006, and 5,036,006, which arewhich both are both incorporatedfully incorporated fullybybyreference. reference. Thevaccine The vaccinecancan be be administered administered to atomammal a mammal to elicit to elicit an immune an immune response response in a in a 2023200517 01 mammal.TheThe mammal. mammal mammal canhuman, can be be human, non-human non-human primate, primate, cow, cow, pig, sheep, pig, sheep, goat, goat, antelope, antelope,

bison, water bison, waterbuffalo, buffalo,bovids, bovids,deer, deer,hedgehogs, hedgehogs, elephants, elephants, llama, llama, alpaca, alpaca, mice,mice, rats,rats, or chicken, or chicken,

and preferably and preferablyhuman, human, cow, cow, pig,pig, or chicken. or chicken.

a. Combination a. Combination Treatments Treatments Thepharmaceutical The pharmaceutical compositions, compositions, preferably preferably vaccines, vaccines, can be can be administered administered in in combinationwith combination with oneone or more or more otherother prostate prostate proteins proteins or genes. or genes. The vaccine The vaccine can be can be administeredinincombination administered combinationwithwith proteins proteins or genes or genes encoding encoding adjuvants, adjuvants, which which can can include: include: a-interferon(IFN- -interferon(IFN- ), a), -interferon ß-interferon (IFN-P), (IFN-), -interferon, y-interferon, IL-12,IL-12, IL-15, IL-15, IL-28, IL-28, CTACK, CTACK, TECK,platelet TECK, platelet derived derived growth growth factor factor(PDGF), (PDGF), TNFc, TNF3, TNF, TNFß, GM-CSF, GM-CSF, epidermal epidermal growth growth

factor (EGF), factor (EGF),IL-1, IL-1,IL-2, IL-2,IL-4, IL-4,IL-5, IL-5,IL-6, IL-6,IL-10, IL-10,IL-12, IL-12, IL-18, IL-18, MCP-1, MCP-1, MIP-la, MIP-la, MIP-1p,MIP-Ip, IL- IL 8, RANTES, 8, L-selectin, P-selectin, RANTES, L-selectin, P-selectin, E-selectin, E-selectin,CD34, GlyCAM-1, CD34, MadCAM-1, GlyCAM-1, MadCAM-1, LFA-1, LFA-1,

VLA-1, Mac-1, VLA-1, Mac-1, p150.95, pl50.95, PECAM, PECAM, ICAM-1, ICAM-1, ICAM-2, ICAM-2, ICAM-3, ICAM-3, CD2, CD2, LFA-3, LFA-3, M-CSF, G M-CSF, G-

CSF,IL-4, CSF, IL-4,mutant mutant forms forms of IL-18, of IL-18, CD40, CD40, CD40L,CD40L, vascularvascular growthfibroblast growth factor, factor, fibroblast growth growth factor, IL-7, factor, IL-7, nerve growthfactor, nerve growth factor,vascular vascularendothelial endothelial growth growth factor, factor, Fas,Fas, TNF TNF receptor, receptor, Flt, Flt, Apo-1, p55, Apo-1, p55,WSL-1, WSL-1,DR3, DR3,TRAMP, Apo-3, AIR, TRAMP, Apo-3, AIR, LARD, LARD, NGRF, DR4, DR5, NGRF, DR4, DR5, KILLER, KILLER, TRAIL-R2,TRICK2, TRAIL-R2, TRICK2, DR6, DR6, Caspase Caspase ICE, ICE, Fos,Fos, c-jun, c-jun, Sp-1, Sp-1, Ap-1,Ap-2, Ap-1, Ap-2, p38,p65Rel, p38, p65Rel, MyD88,IRAK, MyD88, IRAK, TRAF6, TRAF6, IkB,IkB, Inactive Inactive NIK, NIK, SAP SAP K, SAP-1, K, SAP-1, JNK, JNK, interferon interferon response response genes, genes,

NFkB, Bax, TRAIL, NFkB, Bax, TRAIL, TRAILrec, TRAILrec, TRAILrecDRC5, TRAIL-R3,TRAIL-R4, TRAILrecDRC5, TRAIL-R3, TRAIL-R4,RANK, RANK, RANK RANK LIGAND, Ox40, LIGAND, Ox40, Ox40 Ox40LIGAND, LIGAND, NKG2D, NKG2D, MICA, MICA, MICB, MICB,NKG2A, NKG2A, NKG2B, NKG2B, NKG2C, NKG2C, NKG2E, NKG2F, NKG2E, NKG2F, TAPI, TAP1, or TAP2, or TAP2, or functional or functional fragments fragments thereof. thereof.

b. Routes b. RoutesofofAdministration Administration Thevaccine The vaccinecancan be be administered administered by different by different routes routes including including orally, orally, parenterally, parenterally,

sublingually, transdermally, sublingually, transdermally,rectally, rectally,transmucosally, transmucosally, topically, topically, viavia inhalation, inhalation, viavia buccal buccal

administration, intrapleurally, administration, intrapleurally, intravenous, intravenous,intraarterial, intraarterial, intraperitoneal, intraperitoneal, subcutaneous, subcutaneous, intramuscular, intranasal intramuscular, intranasalintrathecal, intrathecal,and andintraarticular intraarticularororcombinations combinations thereof. thereof. ForFor

veterinary use, veterinary use, the the composition compositioncancan be be administered administered as a as a suitably suitably acceptable acceptable formulation formulation in in accordancewith accordance with normal normal veterinary veterinary practice. practice. The The veterinarian veterinarian can readily can readily determine determine the the dosingregimen dosing regimenandand route route of administration of administration that that is most is most appropriate appropriate for a for a particular particular animal.. animal..

Thevaccine The vaccinecancan be be administered administered by traditional by traditional syringes, syringes, needleless needleless injection injection devices, devices,

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"microprojectilebombardment "microprojectile bombardmentgone gone guns",guns", or physical or other other physical methodsmethods such as electroporation such as electroporation

("EP"), "hydrodynamic ("EP"), "hydrodynamic method", method", or ultrasound. or ultrasound.

Thevector The vectorofofthe thevaccine vaccinecancan be be delivered delivered to the to the mammal mammal by several by several well well known known technologiesincluding technologies includingDNADNA injection injection (also(also referred referred to astoDNA as vaccination) DNA vaccination) with andwith and without without in vivo in electroporation, liposome vivo electroporation, liposome mediated, mediated, nanoparticle nanoparticle facilitated, facilitated, recombinant recombinant vectors vectors such such as recombinant as adenovirus, recombinant adenovirus, recombinant recombinant adenovirus adenovirus associated associated virus virus and and recombinant recombinant

vaccinia. The vaccinia. Theprostate prostateantigen antigencancan be be delivered delivered via via DNA DNA injection injection and with and along alonginwith vivo in vivo electroporation. electroporation.

c. Electroporation c. Electroporation Administration Administration of of thevaccine the vaccine viavia electroporation electroporation of the of the plasmids plasmids ofvaccine of the the vaccine may may be accomplished be accomplished using using electroporation electroporation devices devices that that canconfigured can be be configured to deliver to deliver to a desired to a desired

tissue of tissue of aa mammal a pulse mammal a pulse of energy of energy effective effective to cause to cause reversible reversible porespores to form to form in in cell cell membranes, membranes, andand in some in some embodiments, embodiments, theofpulse the pulse ofis energy energy is a constant a constant current tosimilar current similar a to a preset current preset current input inputby bya auser. user. In some In embodiments some embodiments wherewhere electroporation electroporation is utilized, is utilized, the electroporation the electroporation device device maycomprise may comprisean an electroporation electroporation component component and an and an electrode electrode assemblyassembly or handle or handle assembly. assembly. Theelectroporation The electroporationcomponent component may include may include and incorporate and incorporate one of one or more or the more of the various various elementsofofthe elements theelectroporation electroporationdevices, devices, including: including: controller, controller, current current waveform waveform generator, generator,

impedancetester, impedance tester,waveform waveform logger, logger, inputinput element, element, statusstatus reporting reporting element, element, communication communication

port, memory port, component,power memory component, powersource, source, and andpower powerswitch. switch. The Theelectroporation electroporation may maybebe accomplished using accomplished using an an in in vivo vivo electroporation electroporationdevice, forfor device, example CELLECTRA@ example CELLECTRA® EP EP

system(Inovio system (InovioPharmaceuticals, Pharmaceuticals, Inc., Inc., BlueBlue Bell, Bell, PA) PA) or Elgen or Elgen electroporator electroporator (Inovio(Inovio

Pharmaceuticals,Inc., Pharmaceuticals, Inc.,Blue Blue Bell,PA)PA) Bell, to facilitatetransfection to facilitate transfectionof of cellsbybythethe cells plasmid. plasmid.

Theelectroporation The electroporationcomponent component may function may function as one as one element element of the electroporation of the electroporation

devices, and devices, andthe theother otherelements elementsareare separate separate elements elements (or components) (or components) in communication in communication

with the with the electroporation electroporationcomponent. component. The The electroporation electroporation component component may as may function function more as more than one than oneelement elementof of theelectroporation the electroporation devices, devices, which which may may be in be in communication communication with stillwith still other elements other elementsofofthe theelectroporation electroporationdevices devices separate separate fromfrom the electroporation the electroporation component. component.

Theelements The elementsof of theelectroporation the electroporation devices devices existing existing as parts as parts of one of one electromechanical electromechanical or or mechanicaldevice mechanical device maymay not limited not limited as elements as the the elements can function can function as one as one or device device or as separate as separate

elements in elements in communication with one communication with one another. another. The The electroporation electroporationcomponent component may be capable may be capable of delivering of the pulse delivering the pulseofofenergy energythat thatproduces producesthethe constant constant current current in the in the desired desired tissue, tissue, and and

includes aa feedback includes feedbackmechanism. mechanism. The electrode The electrode assembly assembly mayaninclude may include an array electrode electrode array havinga aplurality having plurality ofofelectrodes electrodesininaaspatial spatial arrangement, arrangement,wherein wherein the the electrode electrode assembly assembly

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Feb 2023 receives the receives the pulse pulseofofenergy energyfrom from thethe electroporation electroporation component component and delivers and delivers same tosame the to the desired tissue desired tissue through throughthe theelectrodes. electrodes.AtAt leastoneone least of of thethe plurality plurality of of electrodes electrodes is is neutral neutral

deliveryofofthe during delivery during thepulse pulseofofenergy energyandand measures measures impedance impedance in the in the desired desired tissue tissue and and communicatesthe communicates theimpedance impedancetotothe the electroporation electroporation component. The feedback component. The feedback mechanism mechanism 2023200517 01 mayreceive may receivethethemeasured measured impedance impedance and and can can the adjust adjust theofpulse pulse of delivered energy energy delivered by the by the electroporation component electroporation component to maintain to maintain the constant the constant current. current.

A plurality of A plurality of electrodes electrodes may maydeliver deliver thethe pulse pulse of of energy energy in aindecentralized a decentralized pattern. pattern.

Theplurality The plurality ofofelectrodes electrodesmay may deliver deliver thethe pulse pulse of of energy energy in the in the decentralized decentralized pattern pattern

throughthe through thecontrol controlofofthe theelectrodes electrodesunder under a programmed a programmed sequence, sequence, and the and the programmed programmed

sequenceisisinput sequence inputbybya auser usertotothe theelectroporation electroporationcomponent. component. The programmed The programmed sequence sequence maycomprise may comprise a plurality a plurality of of pulses pulses delivered delivered in sequence, in sequence, wherein wherein each of each pulse pulse the of the plurality plurality

of pulses of is delivered pulses is byatat least delivered by least two two active active electrodes electrodeswith withoneone neutral neutral electrode electrode that that

measuresimpedance, measures impedance, and and wherein wherein a subsequent a subsequent pulse pulse of of the plurality the plurality of pulses of pulses is delivered is delivered

by aa different by different one oneofofatat least least two active electrodes two active electrodeswith withone oneneutral neutral electrode electrode that that measures measures

impedance. impedance.

The feedback The feedback mechanism mechanismmay may be be performed performed by by eitherhardware either hardwareororsoftware. software. The The feedback mechanism feedback mechanismmay maybe be performed performed by by an an analogclosed-loop analog closed-loopcircuit. circuit. The The feedback feedback occurs every occurs every5050µs,ps,2020µs,ps,10 10 µs ps or or 1 1 µs, s, butbut is is preferably preferably a real-time a real-time feedback feedback or or instantaneous(i.e., instantaneous (i.e., substantially substantially instantaneous instantaneousasasdetermined determined by available by available techniques techniques for for determiningresponse determining response time). time). TheThe neutral neutral electrode electrode may measure may measure the impedance the impedance in the in the desired desired tissue and tissue and communicates the impedance communicates the to the impedance to the feedback feedback mechanism, and the mechanism, and the feedback feedback

mechanism mechanism responds responds to impedance to the the impedance and adjusts and adjusts theofpulse the pulse of to energy energy to maintain maintain the the constant current constant currentatat aa value valuesimilar similartoto the the preset preset current. current. The The feedback feedback mechanism mechanism may may maintainthe maintain theconstant constantcurrent currentcontinuously continuously and and instantaneously instantaneously duringduring the delivery the delivery of the of the pulse of pulse ofenergy. energy. Examples Examples of of electroporation electroporation devices devices and and electroporation electroporation methods methods that that may may facilitate facilitate

delivery of delivery of the the DNA DNA vaccines vaccines of the of the present present invention, invention, include include those those described described in U.S.inPatent U.S. Patent No. 7,245,963by by No. 7,245,963 Draghia-Akli, Draghia-Akli, et al., et al., U.S.U.S. Patent Patent Pub.Pub. 2005/0052630 2005/0052630 submitted submitted by Smith,by et Smith, et

al., the al., thecontents contents of of which are hereby which are herebyincorporated incorporated by by reference reference in their in their entirety. entirety. Other Other

electroporation devices electroporation devicesandand electroporation electroporation methods methods that bemay that may befor used used for facilitating facilitating delivery delivery

of the of the DNA vaccines DNA vaccines include include those those provided provided in co-pending in co-pending and co-owned and co-owned U.S. U.S. Patent Patent Application,Serial Application, SerialNo. No.11/874072, 11/874072, filed filed October October 17, 2007, 17, 2007, which which claims claims the benefit the benefit under under 35 35 USC119(e) USC 119(e) to to U.S. U.S. Provisional Provisional Applications Applications Ser. Nos. Ser. Nos. 60/852,149, 60/852,149, filed October filed October 17, 2006,17, 2006, and 60/978,982, and 60/978,982,filed filedOctober October 10, 10, 2007, 2007, all all of which of which are hereby are hereby incorporated incorporated in entirety. in their their entirety.

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Feb 2023 U.S. No.7,245,963 PatentNo. U.S. Patent by by 7,245,963 Draghia-Akli, Draghia-Akli, et describes et al. al. describes modular modular electrode electrode

systemsand systems andtheir theiruse usefor forfacilitating theintroduction facilitating the introductionofofa abiomolecule biomoleculeintointo cells cells of of a selected a selected

tissue in tissue in aa body or plant. body or plant. The Themodular modular electrode electrode systems systems may comprise may comprise a plurality a plurality of of needle needle electrodes; aa hypodermic electrodes; hypodermic needle; needle; an electrical an electrical connector connector that that provides provides a conductive a conductive link link from from 2023200517 01 a programmable a programmable constant-current constant-current pulsepulse controller controller to plurality to the the plurality of needle of needle electrodes; electrodes; and a and a powersource. power source.AnAn operator operator can can grasp grasp the plurality the plurality of needle of needle electrodes electrodes thatmounted that are are mounted on a on a support structure support structure and andfirmly firmlyinsert insertthem them into into thethe selected selected tissue tissue in in a body a body or plant. or plant. TheThe

biomoleculesarearethen biomolecules then delivered delivered viavia thethe hypodermic hypodermic needleneedle intoselected into the the selected tissue.tissue. The The programmable programmable constant-current constant-current pulsepulse controller controller is activated is activated and constant-current and constant-current electrical electrical

pulse is pulse is applied to the applied to the plurality plurality of of needle needle electrodes. electrodes. The Theapplied applied constant-current constant-current electrical electrical

pulse facilitates pulse facilitates the the introduction of the introduction of the biomolecule biomoleculeinto intothethecell cellbetween betweenthe the plurality plurality of of electrodes. The electrodes. Theentire entirecontent contentofofU.S. U.S.Patent Patent No.No. 7,245,963 7,245,963 is hereby is hereby incorporated incorporated by by reference. reference.

U.S. PatentPub. U.S. Patent Pub.2005/0052630 2005/0052630 submitted submitted by Smith, by Smith, et al. describes et al. describes an electroporation an electroporation

device which device whichmaymay be used be used to effectively to effectively facilitate facilitate the the introduction introduction of aof a biomolecule biomolecule into into cells cells of aa selected of tissue in selected tissue in aa body or plant. body or plant. The Theelectroporation electroporationdevice device comprises comprises an electro-kinetic an electro-kinetic

device ("EKD device ("EKD device") device") whose whose operation operation is specified is specified by software by software or firmware. or firmware. The EKD The EKD device produces device producesa series a seriesofofprogrammable programmable constant-current constant-current pulse patterns pulse patterns betweenbetween electrodes electrodes

in an in array based an array basedononuser usercontrol controlandand input input of of thethe pulse pulse parameters, parameters, and and allows allows the storage the storage

and acquisition and acquisitionofofcurrent currentwaveform waveform data. data. The The electroporation electroporation devicedevice also comprises also comprises a a replaceable electrode replaceable electrodedisk diskhaving having an an array array of needle of needle electrodes, electrodes, a central a central injection injection channel channel for for an injection an injection needle, needle, and anda aremovable removable guide guide disk. disk. The The entire entire content content of U.S. of U.S. PatentPatent Pub. Pub. 2005/0052630 2005/0052630 is hereby is hereby incorporated incorporated by reference. by reference.

Theelectrode The electrodearrays arraysandand methods methods described described in U.S. in U.S. Patent Patent No. 7,245,963 No. 7,245,963 and U.S.and U.S. Patent Pub. Patent Pub.2005/0052630 2005/0052630 may may be be adapted adapted forpenetration for deep deep penetration into not into onlynot only such tissues tissues as such as muscle,but muscle, butalso alsoother othertissues tissuesorororgans. organs.Because Because of the of the configuration configuration of electrode of the the electrode array, array,

the injection the injection needle needle (to (to deliver deliver the the biomolecule biomoleculeof of choice) choice) is is also also inserted inserted completely completely into into the the target organ, target andthe organ, and theinjection injectionisis administered administeredperpendicular perpendicular to the to the target target issue, issue, in in thethe area area

that is that is pre-delineated by the pre-delineated by the electrodes electrodesThe Theelectrodes electrodes described described in U.S. in U.S. Patent Patent No. 7,245,963 No. 7,245,963

and U.S. and U.S. Patent PatentPub. Pub.2005/005263 2005/005263 are preferably are preferably 20 mm 20 mm long andlong and 21 21 gauge. gauge. Additionally, contemplated Additionally, contemplated in some in some embodiments embodiments that incorporate that incorporate electroporation electroporation

devices and devices anduses usesthereof, thereof,there thereare areelectroporation electroporation devices devices thatthat areare those those described described in in the the followingpatents: following patents:USUSPatent Patent 5,273,525 5,273,525 issued issued December December 28,US1993, 28, 1993, US6,110,161 Patents Patents 6,110,161 issued August issued August29,29,2000, 2000, 6,261,281 6,261,281 issued issued July July 17, 2001, 17, 2001, and 6,958,060 and 6,958,060 issued October issued October 25, 25,

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Feb 2023 2005, and 2005, andUSUS patent patent 6,939,862 6,939,862 issued issued September September 6, 2005. 2005. Furthermore, 6, Furthermore, patents patents covering covering subject matter subject matterprovided providedin inUSUS patent patent 6,697,669 6,697,669 issued issued February February 24, which 24, 2004, 2004,concerns which concerns delivery of delivery ofDNA DNA using using any any of a of a variety variety of devices, of devices, andpatent and US US patent 7,328,064 7,328,064 issued February issued February

5, 2008, 5, drawntotomethod 2008, drawn method of injecting of injecting DNA DNA are contemplated are contemplated herein. herein. The above-patents The above-patents are are 2023200517 01 incorporatedbybyreference incorporated referencein intheir theirentirety. entirety.Another Another embodiment embodiment of an electroporation of an electroporation device device to be to used with be used withthe thecancer cancerantigens antigens described described herein herein is the is the Elgen Elgen EP device EP device (Inovio (Inovio

Pharmaceuticals,Inc., Pharmaceuticals, Inc.,Blue Blue Bell,PA). Bell, PA). d. Method d. MethodofofPreparing Preparing Vaccine Vaccine

Provided herein Provided herein is ismethods methods for forpreparing preparingthe DNA the DNA plasmids plasmids that thatcomprise comprisethe DNA the DNA

vaccinesdiscussed vaccines discussedherein. herein.TheThe DNA DNA plasmids, plasmids, after after the the subcloning final final subcloning step step into theinto the mammalian mammalian expression expression plasmid, plasmid, can becan betoused used to inoculate inoculate a cell culture a cell culture in a scale in a large large scale fermentationtank, fermentation tank,using usingknown known methods methods in theinart. the art. TheDNA The DNA plasmids plasmids for with for use use with thedevices the EP EP devices of the of the present present invention invention can be can be formulatedorormanufactured formulated manufactured using using a combination a combination of devices of known known and devices and techniques, techniques, but but preferably they preferably theyare aremanufactured manufactured using using an optimized an optimized plasmid plasmid manufacturing manufacturing technique technique that is that is describedininaalicensed, described licensed, co-pending co-pending U.S. U.S. provisional provisional application application U.S. U.S. Serial Serial No. 60/939,792, No. 60/939,792,

whichwas which wasfiled filedononMayMay 23, 23, 2007. 2007. In some In some examples, examples, the DNA the DNA used plasmids plasmids used in these in these studies studies can be can be formulated formulatedat atconcentrations concentrations greater greater thanthan or equal or equal to mg/mL. to 10 10 mg/mL. The manufacturing The manufacturing

techniquesalso techniques alsoinclude includeororincorporate incorporate various various devices devices and protocols and protocols thatcommonly that are are commonly known known toto those those of of ordinary ordinary skillin inthetheart, skill art,ininaddition additiontotothose thosedescribed described in in U.S.Serial U.S. Serial No.No.

60/939792,including 60/939792, including those those described described in ain a licensed licensed patent, patent, US Patent US Patent No. 7,238,522, No. 7,238,522, which which issued on issued onJuly July3,3, 2007. 2007.TheThe above-referenced above-referenced application application and patent, and patent, US No. US Serial Serial No. 60/939,792andand 60/939,792 US US Patent Patent No. No. 7,238,522, 7,238,522, respectively, respectively, are hereby are hereby incorporated incorporated in their in their entirety. entirety.

EXAMPLES EXAMPLES Thepresent The presentinvention inventionis isfurther furtherillustrated illustratedininthe the following followingExamples. Examples. It should It should be be understoodthat understood thatthese theseExamples, Examples, while while indicating indicating preferred preferred embodiments embodiments of the invention, of the invention, are are given by given byway wayof of illustrationonly. illustration only.From From thethe above above discussion discussion and these and these Examples, Examples, one one skilled skilled in the in the art art can can ascertain ascertain the the essential essential characteristics characteristics of of this this invention, invention, and withoutdeparting and without departing fromthe from thespirit spirit and and scope scopethereof, thereof,can canmake make various various changes changes and modifications and modifications of the of the invention toto adapt invention adaptitit to to various various usages usagesand andconditions. conditions. Thus, Thus, various various modifications modifications of theof the invention inin addition invention additiontotothose thoseshown shownand and described described herein herein will will be apparent be apparent to those to those skilled skilled in in

-39- the art the art from the foregoing from the description.Such foregoingdescription. Such modifications modifications are also are also intended intended to fall to fall within within the the scope ofofthe scope appended theappended claims. claims.

2023200517 01 Feb

Example 11 Example

Consensus immunogens Consensus immunogensforfor PSA PSA andand PSMA PSMA were were designed designed from from the available the available full full-

length human length and macaque human and macaquesequences sequencesininthe the GenBank GenBank databaseasaspreviously database previously described described in in Laddy,D.J., Laddy, D.J., Yan, Yan,J.,J.,Corbitt, Corbitt, N., N., Kobasa, Kobasa,D.,D.,Kobinger, Kobinger, G.P., G.P., Weiner, Weiner, D.B. (2007). D.B. (2007).

Immunogenicityofofnovel Immunogenicity novel consensus-based consensus-basedDNA DNA vaccines vaccines againstavian against avianinfluenza. influenza. Vaccine. Vaccine. 25,2984-2989,andand 25,2984-2989, Laddy, Laddy, D.J.,D.J., Yan,Yan, J., Kutzler, J., Kutzler, M., Kobasa, M., Kobasa, D., Kobinger, D., Kobinger, G.P.,A.S., G.P., Khan, Khan, A.S., Greenhouse,J.,J.,Sardesai, Greenhouse, Sardesai,N.Y., N.Y., Draghia-Akli, Draghia-Akli, R., Weiner, R., Weiner, D.B. (2008). D.B. (2008). Heterosubtypic Heterosubtypic

Protection against Protection againstPathogenic Pathogenic Human Human and Avian and Avian Influenza Influenza Viruses Viruses via via In Vivo In Vivo Electroporation of Electroporation ofSynthetic SyntheticConsensus ConsensusDNA Antigens. PLoS DNA Antigens. PLoSONE. ONE. 3,e2517. 3,e2517.

Theconsensus The consensus antigen antigen sequences sequences were were synthesized synthesized by GeneScript by GeneScript (Piscataway, (Piscataway, NJ). NJ). AnHAHA An tagtag waswas included included in C-terminus in the the C-terminus of theof the antigen antigen sequence. sequence. Thesequences The antigen antigen sequences were optimized were optimized for for mRNA stability and mRNA stability and codon codon usage usage in in humans. Thefinal humans. The final sequences sequences were were clonedinin the cloned the BamHI BamHIand and XholXhoI sitessites of pVAX1 of the the pVAX Ivector vector (Invitrogen, (Invitrogen, Carlsbad,Carlsbad, CA). CA). A consensus PSA A consensus PSAantigen antigen11 (SEQ (SEQIDIDNO:2) NO:2) was was generated.This generated. This sequence,which sequence, which comprises261 comprises 261amino amino acids, acids, was was compared compared to eachtoofeach of the the PSA PSA sequences sequences setTable set forth in forth 1. in Table 1. The PSA The PSAsequences sequencesused usedinclude include two twohuman humansequences, sequences,a asequence sequencefrom fromM.M.fascicularis, fascicularis, and and

a sequence a sequence from from M. mulatta. Table M. mulatta. Table 11 includes includes the theSEQ SEQ ID ID NO: and Accession NO: and Accession number numberfor for each sequence each used in sequence used in the the comparison comparison with with consensus consensus PSA antigen 11 (SEQ PSA antigen ID NO:2). (SEQ ID NO:2). Table 11 Table

SEQ SEQ Speciesand Species andprotein protein Accession Accession Number Number Number of Number of %homology % homology ID NO ID NO amino acids amino acids to SEQ to ID SEQ ID NO:2 NO:2 17 17 H. sapiens H. sapiensPSA PSA isol isol NP001639.1 NP001639.1 261 261 91 91

18 18 H. sapiens H. sapiens PSA PSA gbAAA60193.1 gbAAA60193.1 262 262 91 91

19 19 M. fascicularis M. fascicularisKLK3 KLK3 Q6DT45.1 Q6DT45.1 261 261 95 95 20 20 M. mulatta M. mulatta PSA PSA NP001036241.1 pp NP001036241.1 261 261 96 96

A multiple sequence A multiple alignment of sequence alignment of H. H. Sapiens Sapiens (SEQ ID NO:17 (SEQ ID NO:17and andSEQ SEQID ID NO:18), NO:18),

M. mulatta (SEQ M. mulatta IDNO:20) (SEQ ID NO:20)and facicularis(SEQ andM.M.facicularis (SEQIDIDNO:19) NO:19) PSA PSA sequences sequences waswas

generatedwith generated withthe theconsensus consensus PSA PSA antigen antigen 1 (SEQ1 ID (SEQ ID KLK3 NO:2). NO:2). KLK3 (kallikrein (kallikrein 3) is the 3) is the gene encoding gene encoding PSA PSAand andisis pseudonymous pseudonymouswith withPSA. PSA. TheThe PSA PSA antigen antigen 1 is1 91% is 91% homologous homologous

to H. to sapiens,96% H. sapiens, 96% homologous homologous to M. to M. mulatta mulatta and 95%and 95% homologous homologous facicularisfull M. full- M. facicularis

length PSA length PSAprotein protein sequences. sequences.

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Example 22 Example

A consensus PSA A consensus (SEQIDIDNO:4) antigen22 (SEQ PSAantigen NO:4) was was generated. This generated.This sequence, which sequence,which comprises279 comprises 279 amino amino acids acids including including anleader an IgE IgE leader sequence, sequence, was compared was compared to each of to each the of the PSAsequences PSA sequencesset set forth forth in inTable Table2.2.The ThePSA used include PSA sequences used include two two human sequences, human sequences,

a sequence a sequencefrom fromM. M.fascicularis, fascicularis, andand a sequence a sequence from from M. mulatta. M. mulatta. Table 2 Table 2 includes includes the SEQ the SEQ ID NO: ID and Accession NO: and Accessionnumber numberfor foreach eachsequence sequenceused usedinin the the comparison with consensus comparison with consensus PSAantigen PSA antigen 22 (SEQ (SEQIDIDNO:4). NO:4). Table 22 Table

SEQ SEQ Speciesand Species andprotein protein Accession Accession Number Number Number of Number of % homology % homology ID NO ID NO amino acids amino acids to SEQ to ID SEQ ID NO:4 NO:4 17 17 H. sapiens H. PSA sapiensPSA isol isol NP001639.1 NP001639.1 261 261 91 91

18 18 H. sapiens H. sapiens PSA PSA gbAAA60193.1 gbAAA60193.1 262 262 90 90 19 19 M. fascicularis M. fascicularisKLK3 KLK3 Q6DT45.1 Q6DT45.1 261 261 95 95 21 21 M. mulatta M. mulatta PSA PSA AAZ82258.1 AAZ82258.1 244 244 95 95

A multiple sequence A multiple sequence alignment of H. H. Sapiens Sapiens (SEQ ID NO:17 (SEQ ID NO:17and andSEQ SEQID ID NO:18), NO:18),

M. mulatta (SEQ M. mulatta IDNO:21) (SEQ ID NO:21)and facicularis(SEQ andM.M.facicularis (SEQIDIDNO:19) NO:19) PSA PSA sequences sequences waswas

generatedwith generated withthe theconsensus consensus PSA PSA antigen antigen 1 (SEQ1 ID (SEQ ID KLK3 NO:4). NO:4). KLK3 (kallikrein (kallikrein 3) is 3) is the gene the gene encoding PSA encoding PSAand andisis pseudonymous pseudonymouswith withPSA. PSA. TheThe PSA PSA antigen antigen 1 is1 90-91% is 90-91% homologous homologous

to H. to sapiensand H. sapiens 95% and95% homologous homologous M.facicularis M. facicularis full-length full-length PSA sequences, PSA protein protein sequences, and and 95%homologous 95% homologousto to M.M.mulatta mulatta partial PSA partial PSAprotein protein sequence. sequence.

Example 33 Example A consensus PSMA A consensus PSMA antigen antigen 1 1 (SEQ (SEQ ID ID NO:6) NO:6) was was generated. generated. ThisThis sequence, sequence, which which

comprises750 comprises 750 amino amino acids acids was was compared compared to eachto ofeach of the the PSMA PSMA set sequences sequences forth inset forth Table in Table 3. The 3. PSMA The PSMA sequences sequences used used includetwo include two human human sequences sequences andand a sequence a sequence from from M. M. mulatta. Table mulatta. Table 33 includes includes the theSEQ SEQ ID ID NO: and Accession NO: and Accession number numberfor foreach each sequence sequenceused used in in the comparison the with consensus comparison with consensus PSMA antigen1 1(SEQ PSMA antigen (SEQIDID NO:6). NO:6).

Table 33 Table

SEQ SEQ Species and Species andprotein protein Accession Number Accession Number Number of Number of % homology % homology ID NO ID NO amino acids amino acids to SEQ to ID SEQ ID NO:6 NO:6 22 22 H. sapiens H. sapiens PSMA GCPII-isol PSMA GCPII_isol NP_004467.1 NP_004467.1 750 750 96 96

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23 H. sapiens H. sapiensPSMA PSMA AAC83972.1 AAC83972.1 749 749 96 96 24 24 M. mulatta M. mulatta GCPII GCPIIisol isol XP_001096141.2 XP_001096141.2 735 735 94 94

A multiple sequence A multiple sequence alignment alignment of of H. H. Sapiens Sapiens and and M. M. mulatta mulatta PSMA sequenceswas PSMA sequences was generated with generated with PSMA antigen1.1. The PSMA antigen ThePSMA PSMA antigen antigen 1 consensus 1 consensus sequence sequence (SEQ (SEQ ID NO:6) ID NO:6)

is is 96% homologoustoto H. 96% homologous H. sapiens sapiensPSMA PSMA (SEQ (SEQ ID ID NO:22 NO:22 and ID and SEQ SEQ ID NO:23) NO:23) and 94% and 94%

homologoustotoM. homologous M.mulatta mulattafull-length full-length PSMA protein sequence PSMA protein sequence (SEQ (SEQIDIDNO:24). NO:24).

Example 44 Example A consensus PSMA A consensus PSMA antigen2 2(SEQ antigen (SEQ ID ID NO:8) NO:8) was was generated. generated. ThisThis sequence, sequence, which which

comprises767767 comprises amino amino acidsacids including including an IgEanleader IgE leader sequence, sequence, was compared was compared to each of to theeach of the PSMA PSMA sequences sequences setsetforth forth in in Table Table 4. 4. The PSMA The PSMA sequencesused sequences used includetwo include twohuman human sequences and sequences and aa sequence from M. sequence from M. mulatta. mulatta. Table Table 44 includes includes the theSEQ SEQ ID ID NO: and Accession NO: and Accession numberfor number for each each sequence sequence used used in in the the comparison comparison with with consensus consensus PSMA antigen22(SEQ PSMA antigen (SEQIDID NO:8). NO:8).

Table 44 Table

SEQ SEQ Speciesand Species andprotein protein Accession Number Accession Number Number of Number of %homology % homology ID NO ID NO amino acids amino acids to SEQ to ID SEQ ID NO:8 NO:8 22 22 H. sapiens H. sapiens PSMA GCPII-isol PSMA GCPII_isol NP_004467.1 NP_004467.1 750 750 96 96 23 23 H. sapiens H. sapiensPSMA PSMA AAC83972.1 AAC83972.1 749 749 96 96 24 24 M. mulatta M. mulatta GCPII GCPIIisol isol XP_001096141.2 XP_001096141.2 735 735 94 94 25 25 M. mulatta M. mulatta GCPII GCPIIiso2 iso2 XP_002799784.1 XP_002799784.1 704 704 94 94

A multiple sequence A multiple sequence alignment alignment of of H. H. sapiens sapiens (SEQ (SEQ ID ID NO:22 andSEQ NO:22 and SEQIDID NO:23) NO:23)

and M. and mulatta PSMA M. mulatta sequences(SEQ PSMA sequences (SEQ ID ID NO:24 NO:24 and and SEQ SEQ ID NO:25) ID NO:25) was generated was generated with with PSMA PSMA antigen2.2.The antigen ThePSMA PSMA antigen antigen 2 consensus 2 consensus sequence sequence (SEQ(SEQ ID NO:8) ID NO:8) is is 96% 96% homologoustotoH. homologous H. sapiens sapiensPSMA PSMA proteinsequences protein sequencesand and94% 94% homologous homologous to mulatta to M. M. mulatta PSMA PSMA proteinsequences. protein sequences.

Example 55 Example A consensus STEAP A consensus STEAP antigen1 1(SEQ antigen (SEQ ID ID NO:10) NO:10) was was generated. generated. ThisThis sequence, sequence,

whichcomprises which comprises 339339 amino amino acids acids was compared was compared to the to each of eachSTEAP of the STEAPset sequences sequences forth in set forth in Table5.5. The Table TheSTEAP STEAP sequences sequences used include used include two fulltwo fullhuman length length human sequences, sequences, a full a full length length sequence from sequence from M. M. mulatta mulattaand and two two shorter shorter human sequences. Table human sequences. Table55 includes includes the the SEQ ID SEQ ID

NO: and Accession NO: and Accessionnumber numberfor foreach eachsequence sequenceused usedinin the the comparison with consensus comparison with consensus STEAPantigen STEAP antigen1 1(SEQ (SEQIDID NO:10). NO:10).

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Table 55 Table

SEQ SEQ Species and Species andprotein protein Accession Number Accession Number Number of Number of % homology % homology ID NO ID NO amino acids amino acids to SEQ to ID SEQ ID NO:10 NO:10 26 26 H. sapiens H. sapiens STEAP1 STEAP1 NP_036581.1 NP_036581.1 339 339 99 99 27 27 H. sapiens H. sapiens STEAPI STEAP1 GbEAL24167.1 Gb_EAL24167.1 339 339 99 99 28 28 M. mulatta M. mulatta STEAP1 STEAP1 XP_001103605.1 XP_001103605.1 339 339 98 98 29 29 H. sapiens H. sapiensSTEAP STEAP1ICRA CRA bb EAW93751.1 EAW93751.1 259 259 94 94 30 30 H. sapiens H. sapiens STEAPI isofor STEAP1 isofor EAW93749.1 EAW93749.1 258 258 94 94

A multiple sequence A multiple sequence alignment alignment of of H. H. Sapiens Sapiens and and M. M. mulatta mulatta STEAP sequenceswas STEAP sequences was generated with generated with the theconsensus consensus STEAP antigen 1. STEAP antigen 1. The The STEAP STEAP antigen1 1consensus antigen consensussequence sequence (SEQ ID (SEQ IDNO:10) NO:10)isis99% 99%homologous homologous to to human human full-lengthisoforms full-length isoforms(SEQ (SEQ ID ID NO:26 NO:26 and and SEQIDIDNO:27), SEQ NO:27),94% 94% homologous homologous to shorter to shorter H.H. sapiens sapiens isoforms(SEQ isoforms (SEQ ID ID NO:29 NO:29 and and SEQ SEQ

ID NO:30), ID and 94% NO:30), and 94%homologous homologousto to M.M. mulatta mulatta full-length STEAP1 full-length STEAPIprotein proteinsequence sequence(SEQ (SEQ ID NO:28). ID NO:28).

Example 66 Example

A consensus STEAP A consensus STEAP antigen2 2(SEQ antigen (SEQID ID NO:12) NO:12) was was generated. generated. ThisThis sequence, sequence,

whichcomprises which comprises356356 amino amino acidsacids was compared was compared to the to each of eachSTEAP of the STEAPset sequences sequences forth in set forth in Table6.6. The Table TheSTEAP STEAP sequences sequences used include used include two fulltwo fullhuman length length human sequences, sequences, a full a full length length sequence from sequence from M. M. mulatta mulattaand and two two shorter shorter human sequences. Table human sequences. Table66 includes includes the the SEQ ID SEQ ID

NO: and Accession NO: and Accessionnumber numberfor foreach eachsequence sequenceused usedinin the the comparison with consensus comparison with consensus STEAPantigen STEAP antigen2 2(SEQ (SEQIDID NO:12). NO:12).

Table 66 Table

SEQ SEQ Species and Species andprotein protein Accession AccessionNumber Number Number of Number of % homology % homology ID NO ID NO amino acids amino acids to SEQ to ID SEQ ID NO:12 NO:12 26 26 H. sapiens H. sapiens STEAP1 STEAP1 NP_036581.1 NP_036581.1 339 339 94 94 27 27 H. sapiens H. sapiens STEAPI STEAP1 GbEAL24167.1 Gb_EAL24167.1 339 339 94 94 28 28 M. mulatta M. mulattaSTEAP1 STEAP1 XP_001103605.1 XP_001103605.1 339 339 94 94 29 29 H. sapiens H. sapiensSTEAP ICRA STEAP1 CRA bb EAW93751.1 EAW93751.1 259 259 88 88

30 30 H. sapiens H. sapiens STEAPI isofor STEAP1 isofor EAW93749.1 EAW93749.1 258 258 88 88

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A sequence alignment of H. multiple sequence A multiple H. Sapiens Sapiens and and M. mulatta STEAP M. mulatta STEAP1 Isequences was sequences was

generated with generated with the theconsensus consensus STEAP STEAP1 Iantigen 2. The antigen 2. TheSTEAP1 STEAPantigen Iantigen 2 consensus 2 consensus

sequence (SEQ sequence (SEQIDIDNO:12) 94%homologous NO:12)isis94% homologous to full-lengthhuman to full-length human isoforms(SEQ isoforms (SEQ ID ID NO:26 andSEQ NO:26 and SEQID ID NO:27), NO:27), 88%88% homologous homologous to shorter to shorter H. sapiens H. sapiens isoforms isoforms (SEQ (SEQ ID ID

NO:29 andSEQ NO:29 and SEQID ID NO:30), NO:30), andand 94%94% homologous homologous to M.tomulatta M. mulatta full-length full-length STEAP STEAP1 protein protein

sequences (SEQ sequences (SEQID IDNO:28). NO:28).

Example 77 Example A consensus PSCA A consensus PSCA antigen(SEQ antigen (SEQID ID NO:14) NO:14) waswas generated. generated. ThisThis sequence, sequence, which which

comprises 131 comprises 131 amino amino acids acids included included the the IgE IgE leader leadersequence sequencewas was compared compared to to PSCA PSCA

sequenceset sequence setforth Table7.7.TheThe forthininTable PSCA PSCA sequence sequence used used was waslength a full a full human lengthsequence. human sequence. Table77includes Table includesthe theSEQ SEQ ID NO: ID NO: and Accession and Accession number number for for the sequence the sequence used in theused in the comparison with comparison with consensus consensus PSCA PSCA antigen(SEQ antigen (SEQID ID NO:14). NO:14).

Table 77 Table

SEQ SEQ Speciesand Species andprotein protein Accession AccessionNumber Number Number of Number of % homology % homology ID NO ID NO amino acids amino acids to SEQ to ID SEQ ID NO:14 NO:14 31 31 H. sapiens H. sapiens PSCA PSCA NP_005663.2 NP_005663.2 114 114 87 87

A multiple sequence A multiple sequence alignment of H. H. Sapiens Sapiens PSCA sequence(SEQ PSCA sequence (SEQIDIDNO:31) NO:31) waswas

generated with generated with the theconsensus consensus PSCA antigen (SEQ PSCA antigen (SEQ ID IDNO:14). NO:14).The The PSCA PSCA antigen antigen consensus consensus

sequence is sequence is 87% homologoustotofull-length 87% homologous full-length H. H. sapiens sapiensPSCA. PSCA.

Example 88 Example

In vitro vitro translation performed performed totoconfirm confirmthethe expression expression of the of the PSA PSA and PSMA and PSMA

antigens. The antigens. The TNT® TNT@ Quick Quick Coupled Coupled Transcription/TranslationSystem Transcription/Translation System andand 35S-methionine 35S-methionine

(Promega) were (Promega) were used. used. The ThepVAX pVAX vector vector alone alone (negativecontrol) (negative control) or or pVAX backbone pVAX backbone with with

the PSA the PSA ororPSMA PSMA antigen antigen inserts inserts and 35S-methionine and 35S-methionine was addedwas added to the to themixture reaction reaction mixture accordingtotothe according themanufacturer's manufacturer's instructions. instructions. The The reaction reaction was carried was carried out atout at for 30°C 30°C2 for 2 hours. Labeled hours. Labeledproteins proteinswere were immunoprecipitated immunoprecipitated with anti-HA with anti-HA Affinity Affinity GelSt. Gel (Sigma, (Sigma, St. Louis, MO) Louis, MO) by by rotation rotation overnight overnight in radioimmunoprecipitation in radioimmunoprecipitation assaybuffer assay (RIPA) (RIPA)at buffer 4°C. at 4°C. The immunoprecipitated The immunoprecipitated proteins proteins were electrophoresed on were electrophoresed on aa SDS-PAGE gelthat SDS-PAGE gel thatwas was subsequentlyfixed subsequently fixedandand dried. dried. Expression Expression of the of the 35S-labeled 35S-labeled proteins proteins was detected was detected by by autoradiography.TheThe autoradiography. results results areare shown shown in Figure in Figure 1. 1.

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Example 99 Example Cellular immunogenicity Cellular of the immunogenicity of the PSA and PSMA PSA and PSMAantigens antigenswas wasdetermined determinedbyby

2023200517 01 Feb Interferon-gamma ELISpot. Interferon-gamma ELISpot. Female 44 to Female to 6-week-old BALB/cmice 6-week-old BALB/c micewere were purchasedfrom purchased from Jackson Jackson Laboratories Laboratories

(Bar Harbor, (Bar Harbor,ME). ME).AllAll animals animals werewere housed housed in a temperature-controlled, in a temperature-controlled, light-cycled light-cycled facility facility

at the at the University ofPennsylvania. University of Pennsylvania.Animal Animal carecare was carried was carried out according out according to the to the guidelines guidelines of of the National the Institutes of National Institutes ofHealth Healthand andthetheUniversity University of of Pennsylvania Pennsylvania Institutional Institutional Care Care and and Use Committee. Use Committee. For cellular For cellular immunogenicity immunogenicity studies, studies, 10 20or µg 10 or 20ofpgeach of each antigen antigen was delivered was delivered to the to the tibialis anterior tibialis anteriormuscle of Balb/c muscle of Balb/c mice micebybyintramuscular intramuscular injection injection followed followed by electroporation by electroporation

using the using the CELLECTRA@ adaptive CELLECTRA® adaptive constant constant current current device device (InovioPharmaceuticals, (Inovio Pharmaceuticals,Inc., Inc., BlueBell, Blue Bell, PA). PA).Mice Mice (n=5 (n=5 per per group) group) received received 2 immunizations 2 immunizations at weeksat0 weeks and 2. 0Two and 0.12. Two 0.1 Ampconstant Amp constant current current square-wave square-wave pulsespulses were delivered were delivered through through a triangular a triangular 3-electrode 3-electrode

array consisting array consisting ofof26-gauge 26-gauge solid solid stainlesssteel stainless steelelectrodes. electrodes.Each Each pulse pulse was was 52 milliseconds 52 milliseconds

in length in with aa 11 second length with seconddelay delaybetween between pulses. pulses. The The mice mice received received a totala of total of 2 immunizations 2 immunizations

that were that administered2 weeks were administered 2 weeks apart. apart. MiceMice were humanely were humanely sacrificed sacrificed 1 week 1 week after the after the secondimmunization second immunizationfor for analysis analysis of cellular of cellular and and humoral humoral immuneimmune responses. responses.

Cellular and Cellular andresponses responseswere were assessed assessed 1 week 1 week after after the last the last immunization immunization (week (week 5). 5). ELISpotanalysis ELISpot analysiswaswas used used to determine to determine antigen-specific antigen-specific secretion secretion of IFNY. of IFNy. Mouse Mouse IFNy IFNy capture antibody capture antibody (R&D Systems,Minneapolis, (R&D Systems, Minneapolis, MN) MN)waswas used used totocoat coatflat-bottom flat-bottom Immobilon- Immobilon P plates P plates (Millipore, (Millipore, Billerica, Billerica, MA) MA)overnight overnight at 4°C. at 4°C. Splenocytes Splenocytes were were aseptically aseptically isolated isolated

and resuspended and resuspended at at in inRI1 R10 media media (Rosewell (Rosewell Park Park Memorial Institute medium Memorial Institute 1640 with medium 1640 with supplemented with supplemented with 10% 10%fetal fetal bovine bovine serum, serum, 1% 1%antibiotic-antimycotic antibiotic-antimycotic and and 0.1 %2 0.1%2-

mercaptoethanol).2x10 mercaptoethanol). 2x10 5 splenocytes splenocytes from immunized from immunized mice were mice addedwere in toadded in toofeach each well the well of the 96-wellplate 96-well plateand andstimulated stimulatedovernight overnight at 37° at 37° C, CO2, C, 5% 5% C02, in the in the presence presence of R10 of R1 (negative (negative

control), concanavalin control), concanavalin A A (positive (positive control) control) (Sigma, (Sigma, St. St. Louis, Louis, MO) MO) or antigen-specific or antigen-specific

peptide pools. peptide pools.The Thenext nextday, day,mouse mouse IFNy IFNy detection detectionantibody antibody(R&D Systems, Minneapolis, (R&D Systems, Minneapolis, MN)waswas MN) added added to the to the plates plates thatthat werewere then then incubated incubated overnight overnight at 4°C.atThe 4°C. The following following day, day, streptavidin-ALP streptavidin-ALP (MabTech, (MabTech, Sweden) Sweden) wastoadded was added to thefor the plates plates for and 2 hours 2 hours and antigen- antigen specific spots specific spotswere werevisualized with visualized BCIP/NPT with BCIP/NPT substrate substrate(MabTech, (MabTech, Sweden). Sweden). PSA and PSA and

PSMA PSMA peptides peptides werewere 15-mer 15-mer peptides peptides spanning spanning thelength the entire entire oflength of the consensus the consensus

immunogen, immunogen, not not including including thetag the HA HAortag or leader leader sequence, sequence, overlapping overlapping by 11 by 11 amino amino acids, acids, and were and were synthesized synthesized by by GenScript GenScript (Piscataway, (Piscataway, NJ). NJ).PSA PSA and and PSMA peptideswere PSMA peptides wereused usedatat a final a final concentration of1.0 concentration of 1.0 µg/mL pg/mLforfor each each peptide. peptide. IFNyIFNy ELISpot ELISpot wasto used was used to evaluate evaluate

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Feb 2023 antigen-specific cellular antigen-specific cellular responses responses1 week 1 week after after thethe last last immunization. immunization. For IFNy For PSA, PSA, IFNy responseswere responses weresimilar similar forfor thethe 10 10 µg pg (772.2 (772.2 +/- +/- 138.138. 2 SFU) 2 SFU) and and 20 µg 20 pg (771.1 (771.1 +/-SFU) +/- 155.2 155.2 SFU) vaccinedoses vaccine doses(Figure (Figure 2A). 2A). In contrast, In contrast, there there was was a dose-dependant a dose-dependant increase increase in in PSMA- PSMA specific FNy specific FNyresponses responses with with 20 microgram 20 microgram of theof the vaccine vaccine (1585.0(1585.0 +/-SFU) +/- 194.0 194.0 as SFU) as

2023200517 01 compared compared to to 10 10 µg pg of the of the vaccine vaccine (1047.2 (1047.2 +/- 160.7 +/- 160.7 SFU) (Figure SFU) (Figure 2B). Minimal 2B). Minimal backgroundbackground

was observed was observed for for PSA or PSMA PSA or PSMA responsesininnaïve responses navemice. mice.

Example 10 Example 10

Vaccine-inducedCD4+ Vaccine-induced CD4+ and and CD8+ CD8+ T cell T cell production production of ofIFN; IL-2 IFN, IL-2 andand TNFTNFa

Cellular immunogenicity Cellular immunogenicity was was further further characterized characterized by cytometry by flow flow cytometry for for the co- the co delivery of delivery ofthe thePSA PSA and and PSMA vaccines. Antigen-specific PSMA vaccines. Antigen-specific CD4+ andCD8+ CD4+ and CD8+ T cell T cell

productionofofIFN, production IFNy, IL-2 IL-2 and and TNFc TNF was was determined determined for the for the total total vaccine-specific vaccine-specific response response and the and the PSA PSA and and PSMA PSMA components components of the of the total total vaccine-specific vaccine-specific response response (n=5). (n=5). Cellular immune Cellular immune responses responses werewere also also determined determined by intracellular by intracellular cytokine cytokine stainingstaining

and flow and flowcytometry cytometry using using the the CytoFix/CytoPerm CytoFix/CytoPerm kit per kit per manufacteurer's manufacteurer's instructions instructions (BD (BD Biosciences, San Biosciences, San Diego, Diego, CA). CA). Splenocytes harvested harvested from from immunized micewere immunized mice werewashed washed with PBS with PBSandand then then resuspended resuspended in media in R10 RI media to a concentration to a final final concentration of 107 cells/ml. of 107 cells/ml. Cells Cells wereseeded were seededinin96-well 96-well round round bottom bottom plates plates in a in a volume volume of100pl of 100µl and an additional and an additional 100µl of100pl of RI media R10 media (negative (negative control), control), media media containing containing antigen-specific antigen-specific peptides peptides pools pools or mediaor media containingphorbol containing phorbolmyristate myristate acetate acetate (PMA, (PMA, 10 ng/ml) 10 ng/ml) and ionomycin and ionomycin (250 (250 ng/ml; ng/ml; positive positive control) (Sigma, control) (Sigma,St. St. Louis, Louis,MO) MO)was was addedadded and plates and plates were incubated were incubated at 37°C,at5%37°C, 5% 6C02, CO2, for for 6 hours. All hours. All stimulation stimulationmedia media contained contained 1 pg/pL 1 µg/µL each each of of GolgiPlug GolgiPlug and GolgiStop and GolgiStop (BD (BD Biosciences,San Biosciences, SanDiego, Diego, CA). CA). At end At the the of endtheofincubation the incubation periodperiod plates plates weredown were spun spun anddown and washedtwice washed twice with with PBS. PBS. Cells Cells werewere then then stained stained with awith a violet violet dye dye for for viability viability (LIVE/DEAD (LIVE/DEAD

Violet Viability Violet Viability Dye, Dye,Invitrogen; Invitrogen;Carlsbad, Carlsbad, CA)CA) forminutes for 30 30 minutes at After at 4°C. 4°C. After washingwashing as as abovewith above withPBS, PBS, cells cells were were stained stained externally externally forminutes for 30 30 minutes with anti-CD4 with anti-CD4 PerCPCy5.5 PerCPCy5.5 and and anti-CD8APC anti-CD8 APC at 4°C, at 4°C, followed followed by fixing by fixing and permeabilization. and permeabilization. Anti-CD3Anti-CD3 PE-Cy5, PE-Cy5, anti-IL-2 anti-IL-2 PE, anti-IFNy PE, anti-IFNy AlexaFluor-700 and anti-TNF AlexaFluor-700 and anti-TNFa FITC FITC (BD(BD Biosciences, Biosciences, SanSan Diego, Diego, CA)CA) were were

addedand added andcells cellswere were incubated incubated again again at 4°C at 4°C forminutes. for 30 30 minutes. Cells Cells were agiven were given final awash final wash with PBS with and fixed PBS and fixed in in 1% PFA. 1% PFA.

Co-delivery of the Co-delivery thePSA PSA and and PSMA vaccineinduced PSMA vaccine inducedrobust robustCD4+ CD4+ secretionofofIFNy, secretion IFNy, IL-2 and TNFc. IL-2 The TNF. The percentageofofPSA-specific percentage PSA-specific(0.21%) (0.21%)and andPSMA-specific PSMA-specific (0.24%) (0.24%) IFNy IFNy

producingCD4+ producing CD4+ T cells T cells contributed contributed equally equally to thetototal the total vaccine-specific vaccine-specific CD4+ TCD4+ T cell cell IFNy IFNy

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Feb 2023 response (0.44%) response (Figure 3A). (0.44%) (Figure 3A). PSMA-specific CD4+T T PSMA-specific CD4+ cellsproducing cells producingIL-2 IL-2(1.08%) (1.08%) comprisedthethemajority comprised majority of of thethe totalpercentage total percentage of CD4+ of CD4+ T cells T cells producing producing vaccine-specific vaccine-specific

IL-2 (1.40%) IL-2 (Figure 3B). (1.40%) (Figure 3B). The The percentage percentage of ofPSA PSA (0.31%) (0.31%) and and PSMA (0.29%) PSMA (0.29%) induced induced

CD4+ CD4+ T cellproduction T cell production of TNFc of TNF contributed contributed equally equally to thevaccine-specific to the total total vaccine-specific response response 2023200517 01 (0.60%) (Figure (0.60%) (Figure 3C). 3C). Overall, Overall,CD4+ T cell CD4+ T cell responses responseswere werewell wellbalanced balancedbetween betweenPSA PSA and and

PSMA,with PSMA, withthe theexception exception of of PSMA PSMA inducing inducing themajority the majorityofofthe the vaccine-specific vaccine-specificCD4+ CD4+ TT

cell IL-2 cell production. IL-2 production.

Thevaccine The vaccineinduced induced strong strong antigen-specific antigen-specific CD8+ CD8+ T cell T cell production production of IFNy of andIFNy IL-2 and IL-2 and, to and, to aalesser extent, lesser TNFc. extent, TNF.Both BothPSA PSA (0.70%) (0.70%) and and PSMA (0.67%) PSMA (0.67%) induced induced robustCD8+ robust CD8+ T T cell IFNy cell production.InInfact, IFNy production. fact,vaccine-specific vaccine-specificCD8+ CD8+ T cells T cells secreting secreting of IFNy of IFNy comprised comprised

1.37%ofofthe 1.37% thetotal totalCD8+ CD8+ T cell T cell population population (Figure (Figure 4A). 4A). The vaccine The vaccine also induced also induced a strong a strong CD8+T cell CD8+ T cellIL-2 IL-2 response response (1.54%). (1.54%). Similar Similar to thetoCD4+ the TCD4+ T cell cell IL-2 IL-2 response, response, the percentage the percentage

of PSMA-specific of (1.06%) PSMA-specific (1.06%) CD8+ CD8+ T cellsTsecreting cells secreting IL-2 IL-2 was was approximately approximately 2-fold 2-fold higher thanhigher than PSA-specific(0.47%) PSA-specific (0.47%) (Figure (Figure 4B).4B). The total The total percentage percentage of vaccine-specific of vaccine-specific CD8+ CD8+ T cell T cell production of production of TNFc (0.11%) TNF (0.11%) was was in in responsetotothe response the PSA PSAcomponent componentof of thethevaccine vaccine(Figure (Figure 4C). In 4C). In summary, summary, there there waswas a high a high percentage percentage of vaccine-specific of vaccine-specific CD8+ T CD8+ T cells production cells production of of IFNyand IFNy andIL-2. IL-2.Similar Similar to to CD4+ CD4+ T cell T cell responses, responses, IFNy production IFNy production was balanced was equally equally balanced between PSA between PSAand andPSMA PSMAand and the the magnitude magnitude of the of the IL-2 IL-2 PSMA-specific PSMA-specific response response waswas greater greater

than that than that of of the PSA-specificresponse. the PSA-specific response.

Example 11 Example 11 PSA-specific IgG PSA-specific IgG Seroconversion Seroconversion

Antibody response can Antibody response can play play an an important important role roleinin tumor immunotherapy. tumor immunotherapy. Accordingly Accordingly

wenext we nextexamined examinedthisthis parameter parameter of immune of the the immune response response to the to the PSA PSAbased antigen antigen on based on protein target availability. protein target availability.

To determine To determine PSA-specific PSA-specific sera sera antibody antibody titers, titers,96-well Nunc-Immuno 96-well Nunc-Immuno MaxiSorp MaxiSorp

plates (Nunc, plates (Nunc,Rochester, Rochester,NY)NY) werewere coated coated overnight overnight at 4°C at 4°C1 with with 1 ofg/well µg/well of recombinant recombinant

PSAprotein PSA protein(Fitzgerald (Fitzgerald Industries,Acton, Industries, Acton, MA) MA) diluted diluted in PBS. in PBS. Plates Plates werewith were washed washed with PBS, 0.05% PBS, 0.05%Tween Tween2020 (PBST), (PBST), blocked blocked forfor 1 1hour houratatroom roomtemperature temperaturewith with10% 10% BSA/PBST, BSA/PBST, and and incubated incubated with serial with serial dilutions dilutions of serum of serum from immunized from immunized or naive or naïve animals animals for 11 hour for at room hour at roomtemperature. temperature.Plates Plates werewere then then washed washed 3 times3 with timesPBST with andPBST and goat goat anti- anti mouseIgG mouse IgG(Santa (Santa Cruz, Cruz, Santa Santa Cruz, Cruz, CA) CA) was wasadded addeda adilution dilution of of 1:5,000 1:5,000ininPBST. PBST. Bound Bound

-47- enzymewas enzyme wasdetected detected by by SigmaFAST SigmaFAST 0-phenylenediamine O-phenylenediamine dihydrochloride dihydrochloride (OPD; (OPD; Sigma Sigma-

Aldrich, St. Aldrich, St. Louis, Louis, MO), MO),andand thethe optical optical density density waswas at 450atnm450 determined determined on nm on a a Biotek Biotek (Winooski,VT) (Winooski, VT) plate plate readeras readeras shown shown in Figure in Figure 5B. Endpoint 5B. Endpoint titersdetermined titers were were determined as as previouslydescribed previously described(Frey, (Frey, A. A. et et al.al.1998). 1998).Briefly, Briefly, thethe upper upper prediction prediction limitlimit was was calculated calculated

using the using the Student Studentt-distribution. t-distribution. The Themathematical mathematical formula formula that that defines defines the upper the upper prediction prediction

limit is limit is expressed as the expressed as the standard standarddeviation deviationmultiplied multiplied by by a factor a factor that that waswas based based on on the the numberofofnegative number negative controls controls (n=5) (n=5) and and the confidence the confidence (95%).(95%). level level The endpoint The endpoint titer wastiter was reported asas the reported the reciprocal reciprocal ofofthe thelast last dilution dilution above abovethe theupper upperpredication predication limit. limit.

In addition In addition to to conferring conferringrobust robustcellular-mediated cellular-mediated immunity, immunity, thevaccine the PSA PSA vaccine also induced also inducedstrong strongantigen-specific antigen-specific humoral humoral responses. responses. Antibody Antibody titers titers were determined were determined by by ELISA ELISA in in seraisolated sera isolatedfrom from mice mice one one week week after after the immunization the last last immunization (n=5). (n=5). The The vaccine vaccine inducedananaverage induced average PSA-specific PSA-specific antibody antibody endpoint endpoint titer titer of of 4,427 4,427 (range(range 1581-15,811) 1581-15,811)

(Figure 5A). (Figure 5A). The Thelongevity longevity of of these these responses responses may may be be important important as as well. well.

Example 12 Example 12 Prostate specific Prostate specific antigen antigenamino amino acid acid sequences sequences available available on GenBank on GenBank include include the the following: gbEAW71923._H.sapiens following: klk3_CRAb; gb_EAW71923.1_H.sapiens_klk3_CRAb;

001639.1_H.sapiensPSAiso lpreproprotein; _001639.1_H.sapiens_PSA_iso1_preproprotein,

gbAAA59995.1_H.sapiensPSAprecursor; gbAAA60193._H.sapiensPSA; gb_AAA59995.1_H.sapiens_PSA_precursor; gb_AAA60193.1_H.sapiens_PSA;

gbEAW71933.1_H.sapiens klk3_CRA_l; gb_EAW71933.1_H.sapiens_klk3_CRA_I:

NP_001025218.1_H.sapiensPSAiso3_preproprotein; gbCAD54617._H.sapiens_PSA; NP_001025218.1_H.sapiens_PSA_iso3_preproprotein,gb_CAD54617.1_Hsapiens_PSA;

gbCAD30844.1_H.sapiensPSA; gbAAA59996.1_H.sapiens_PSA-precursor; gb_CAD30844.1_Hsapiens_PSA,gb_AAA59996.1_H.sapiens_PSA_precursor;

gbAAD14185.1_H.sapiensPSA; Q6DT45.1_M.fascicularisKLK3; gb_AAD14185.1_H.sapiens_PSA;Q6DT45.1_M.fascicularis_KLK3;

NP_001036241.1_M.mulattaPSAprecursor; AAZ82258.1_M.mulattaPSA; NP_001036241.1_M.mulatta_PSA_precursor; AAZ82258.1_M.mulatta_PSA;

anubis]; gi|73746696|gb|AAZ82261.1| anubis]; gil73746696|gblAAZ82261.1 prostate prostate specific specific antigenantigen

[Papio [Papio anubis];anubis];

il73746692|gblAAZ82259.l1 i|73746692[gb[AAZ82259.1| prostate prostate specific specific antigen antigen [Erythrocebus

[Erythrocebus patas]; patas]; gil73746694|gblAAZ82260. prostate gi|73746694gb[AAZ82260.1 I prostate specific specific antigen antigen [Cercopithecus

[Cercopithecus cephus]; cephus];

gil73746682|gblAAZ82254. prostate gi|73746682|gb|AAZ82254.1 prostate specific specific antigenantigen [Pan paniscus];

[Pan paniscus];

gil73746680|gblAAZ82253. prostate gi|73746680|gb|AAZ82253.1| prostate specific specific antigen antigen [Pan troglodytes];

[Pan troglodytes];

gil73746686|gblAAZ82256.l prostate specific gi|73746686|gb|AAZ82256.1| prostate specific antigen antigen[Pongo

[Pongo pygmaeus]; pygmaeus]; and and

3746688|gblAAZ82257.1 13746688|gb|AAZ82257.1 prostate prostate specific specific antigen antigen [Nomascus

[Nomascus gabriellae]. gabriellae].

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PSMA PSMA amino amino sequencesavailable acidsequences acid availableononGenBank GenBank include include thethe following: following:

NP_004467.1_HumanGCPIIisol; HumanPSMA_AAC83972.1; NP_004467.1_Human_GCPII_iso1; Human_PSMA_AAC83972.1; M.mulattaGCPII-isol M.mulatta_GCPII_iso1

2023200517 01 Feb XP_001096141.2;andand XP_001096141.2; M.mulattaGCPIIiso2_XP_002799784.1. M.mulatta_GCPII_iso2_XP_002799784.1.

STEAPamino STEAP amino acidsequences acid sequencesavailable availableononGenBank GenBank includethethefollowing: include following: NP036581.1_HumanSTEAPI;EAL24167.1_HumanSTEAPI; NP036581.1_Human_STEAP1; EAL24167.1_Human_STEAP1;

XP001103605.lM.mulattaSTEAPI_iso3; EAW93751.1_Human_STEAP1_CRAb; XP001103605.1_M.mulatta_STEAP1_iso3; EAW93751.lHumanSTEAPICRAb; EAW93749.lHumanSTEAP1_CRAa; EAW93749.1_Human_STEAP1_CRAa; XP001164838.1_P.troglodytesSTEAPiso2; XP001164838.1_P.troglodytes_STEAPiso2; XP002818311.1_P.abeliiSTEAPI; NP001162459.1_P.anubis_STEAP1; XP002818311.1_P.abeli_STEAP1; NP001162459.1_P.anubisSTEAPi; NP_999470.1_S.scrofaSTEAPl; and NP_081675.2_M.musculusSTEAPI. NP_999470.1_S.scrofa_STEAP1; and NP_081675.2_M.musculus_STEAP1.

NP_005663.2_HumanPSCA NP_005663.2_Human_PSCA is theisaccession the accession number number of a of a PSCA PSCA amino amino acid acid sequence available sequence available on on GenBank. GenBank.

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Claims

THE CLAIMS DEFINING THE INVENTION ARE AS FOLLOWS 11 Aug 2025

1. A vaccine comprising a nucleic acid molecule encoding one or more proteins selected from the group consisting of: a) a protein comprising: (i) an amino acid sequence set forth in SEQ ID NO:2; (ii) an amino acid sequence that is at least 95% identical to the sequence set forth in SEQ ID NO:2, provided that amino acids at positions 69, 78, 80, 82, 102, 110, 137, 139, 165, 189, 203, 220, 2023200517

232 and 248 of the sequence set forth in SEQ ID NO:2 are conserved; or (iii) an immunogenic fragment of (i), wherein the immunogenic fragment comprises amino acids corresponding to at least 256 amino acid residues of the sequence set forth in SEQ ID NO:2, provided that amino acids 69, 78, 80, 82, 102, 110, 137, 139, 165, 189, 203, 220, 232 and 248 of the sequence set forth in SEQ ID NO:2 are conserved; b) a protein comprising: (i) an amino acid sequence set forth in SEQ ID NO:4; (ii) an amino acid sequence that is at least 95% identical to the sequence set forth in SEQ ID NO:4, provided that amino acids at positions 21, 86, 127, 129, 154, 156, 182, 195, 206, 218, 220, 237, 249, 255, 265, 271 or 275 of the sequence set forth in SEQ ID NO:4 are conserved; or (iii) or an immunogenic fragment of (i), wherein the immunogenic fragment comprises amino acids corresponding to at least 274 amino acid residues of the sequence set forth in SEQ ID NO:4, provided that amino acids 21, 86, 127, 129, 154, 156, 182, 195, 206, 218, 220, 237, 249, 255, 265, 271 or 275 of the sequence set forth in SEQ ID NO:4 are conserved; c) a protein comprising: (i) an amino acid sequence set forth in SEQ ID NO:6; (ii) an amino acid sequence that is at least 95% identical to the sequence set forth in SEQ ID NO:6, provided that amino acids at positions 14, 15, 32, 47, 58, 79,111, 157, 223, 320, 350, 475, 499, 569, 613, 624, 653, 660, 663, 733 and 734 of the sequence set forth in SEQ ID NO:6 are conserved; or (iii) an immunogenic fragment of (i), wherein the immunogenic fragment comprises amino acids corresponding to at least 735 amino acid residues of the sequence set forth in SEQ ID NO:6, provided that amino acids 14, 15, 32, 47, 58, 79,111, 157, 223, 320, 350, 475, 499, 569, 613, 624, 653, 660, 663, 733 and 734 of the sequence set forth in SEQ ID NO:6 are conserved; d) a protein comprising: (i) an amino acid sequence set forth in SEQ ID NO:8; (ii) an amino acid sequence that is at least 95% identical to the sequence set forth in SEQ ID NO:8, provided that amino acids at positions 20, 30, 31, 48, 63, 74, 95, 127, 173, 239, 336, 366, 491, 515, 564, 585, 629, 640, 669, 676, 679, 749 and 750 of the sequence set forth in SEQ ID NO:8 are conserved; or (iii) an immunogenic fragment of (i), wherein the immunogenic fragment comprises amino acids corresponding to at least 751 amino acid residues of the 11 Aug 2025 sequence set forth in SEQ ID NO:8, provided that amino acids 20, 30, 31, 48, 63, 74, 95, 127, 173, 239, 336, 366, 491, 515, 564, 585, 629, 640, 669, 676, 679, 749 and 750 of the sequence set forth in SEQ ID NO:8 are conserved; e) a protein comprising an amino acid sequence set forth in SEQ ID NO:10; f) a protein comprising (i) an amino acid sequence set forth in SEQ ID NO:12; (ii) an amino acid sequence that is at least 95% identical to the sequence set forth in SEQ ID NO:12; 2023200517 or (iii) an immunogenic fragment of (i), wherein the immunogenic fragment comprises amino acids corresponding to at least 349 amino acid residues of the sequence set forth in SEQ ID NO:12; and g) a protein comprising: (i) an amino acid sequence set forth in SEQ ID NO:14; (ii) an amino acid sequence that is at least 95% identical to the sequence set forth in SEQ ID NO:14; or (iii) an immunogenic fragment of (i), wherein the immunogenic fragment comprises amino acids corresponding to at least 129 amino acid residues of the sequence set forth in SEQ ID NO:14.

2. The vaccine of claim 1, comprising a nucleic acid molecule encoding at least one protein selected from a) or b) and at least one protein selected from c), d), e), f) and g).

3. The vaccine of claim 1, comprising a nucleic acid molecule encoding one or more proteins selected from the group consisting of: a protein comprising the amino acid sequence set forth in SEQ ID NO:2; a protein comprising the amino acid sequence set forth in SEQ ID NO:4; a protein comprising the amino acid sequence set forth in SEQ ID NO:6; a protein comprising the amino acid sequence set forth in SEQ ID NO:8; a protein comprising the amino acid sequence set forth in SEQ ID NO:10; a protein comprising the amino acid sequence set forth in SEQ ID NO:12; and a protein comprising the amino acid sequence set forth in SEQ ID NO:14.

4. The vaccine of claim 1, wherein the nucleic acid molecule comprises one or more sequence selected from the group consisting of: a) a nucleotide sequence set forth in SEQ ID NO:1, or a nucleotide sequence that is at least 95% identical to the sequence set forth in SEQ ID NO:1; b) a nucleotide sequence set forth in SEQ ID NO:3, or a nucleotide sequence that is at least 95% identical to the sequence set forth in SEQ ID NO:3; c) a nucleotide sequence set forth in nucleotides 1-2250 of SEQ ID NO:5, or a 11 Aug 2025 nucleotide sequence that is at least 95% identical to the nucleotide sequence set forth in nucleotides 1-2250 of SEQ ID NO:5; d) a nucleotide sequence set forth in nucleotides 1-2301 of SEQ ID NO:7, or a nucleotide sequence that is at least 95% identical to the nucleotide sequence set forth in nucleotides 1-2301 of SEQ ID NO:7; e) a nucleotide sequence set forth in SEQ ID NO:9; 2023200517 f) a nucleotide sequence set forth in SEQ ID NO:11, or a nucleotide sequence that is at least 95% identical to the sequence set forth in SEQ ID NO:11; and g) a nucleotide sequence set forth in SEQ ID NO:13, or a nucleotide sequence that is at least 95% identical to the sequence set forth in SEQ ID NO:13.

5. The vaccine of claim 4, comprising one or more nucleotide sequences selected from the group consisting of: the nucleotide sequence set forth in SEQ ID NO:1; the nucleotide sequence set forth in SEQ ID NO:3; the nucleotide sequence set forth in nucleotides 1-2250 of SEQ ID NO:5; the nucleotide sequence set forth in nucleotides 1-2301 of SEQ ID NO:7; the nucleotide sequence set forth in SEQ ID NO:9; SEQ ID NO:11; and the nucleotide sequence set forth in SEQ ID NO:13.

6. The vaccine of any one of claims 1-5, wherein the nucleic acid molecule is a plasmid.

7. The vaccine of any one of claims 1-6, wherein the nucleic acid molecule is an expression vector and sequences encoding said one or more proteins are operably linked to one or more regulatory elements.

8. A method of treating an individual who has been diagnosed with prostate cancer, comprising administering the vaccine of any one of claims 1-7 to the individual.

9. Use of the vaccine of any one of claims 1-7 in the preparation of a medicament for treating prostate cancer in an individual who has been diagnosed therewith.

10. A composition for inducing an immune response against a prostate cancer antigen, said composition comprising a protein cancer antigen selected from the group consisting of: a) a protein comprising: (i) an amino acid sequence set forth in SEQ ID NO:2; 11 Aug 2025

(ii) an amino acid sequence that is at least 95% identical to the sequence set forth in SEQ ID NO:2, provided that amino acids at positions 69, 78, 80, 82, 102, 110, 137, 139, 165, 189, 203, 220, 232 and 248 of the sequence set forth in SEQ ID NO:2 are conserved; or (iii) an immunogenic fragment of (i), wherein the immunogenic fragment comprises amino acids corresponding to at least 256 amino acid residues of the sequence set forth in SEQ ID NO:2, provided that amino acids 69, 78, 80, 82, 102, 110, 137, 139, 165, 189, 203, 220, 232 and 248 2023200517

of the sequence set forth in SEQ ID NO:2 are conserved; b) a protein comprising: (i) an amino acid sequence set forth in SEQ ID NO:4; (ii) an amino acid sequence that is at least 95% identical to the sequence set forth in SEQ ID NO:4, provided that amino acids at positions 21, 86, 127, 129, 154, 156, 182, 195, 206, 218, 220, 237, 249, 255, 265, 271 or 275 of the sequence set forth in SEQ ID NO:4 are conserved; or (iii) an immunogenic fragment of (i), wherein the immunogenic fragment comprises amino acids corresponding to at least 274 amino acid residues of the sequence set forth in SEQ ID NO:4, provided that amino acids 21, 86, 127, 129, 154, 156, 182, 195, 206, 218, 220, 237, 249, 255, 265, 271 or 275 of the sequence set forth in SEQ ID NO:4 are conserved; c) a protein comprising: (i) an amino acid sequence set forth in SEQ ID NO:6; (ii) an amino acid sequence that is at least 95% identical to the sequence set forth in SEQ ID NO:6, provided that amino acids at positions 14, 15, 32, 47, 58, 79,111, 157, 223, 320, 350, 475, 499, 569, 613, 624, 653, 660, 663, 733 and 734 of the sequence set forth in SEQ ID NO:6 are conserved; or (iii) an immunogenic fragment of (i), wherein the immunogenic fragment comprises amino acids corresponding to at least 735 amino acid residues of the sequence set forth in SEQ ID NO:6, provided that amino acids 14, 15, 32, 47, 58, 79,111, 157, 223, 320, 350, 475, 499, 569, 613, 624, 653, 660, 663, 733 and 734 of the sequence set forth in SEQ ID NO:6 are conserved; d) a protein comprising: (i) an amino acid sequence set forth in SEQ ID NO:8; an amino acid sequence that is at least 95% identical to the sequence set forth in SEQ ID NO:8, provided that amino acids at positions 20, 30, 31, 48, 63, 74, 95, 127, 173, 239, 336, 366, 491, 515, 564, 585, 629, 640, 669, 676, 679, 749 and 750 of the sequence set forth in SEQ ID NO:8 are conserved, or (iii) an immunogenic fragment of (i), wherein the immunogenic fragment comprises amino acids corresponding to at least 751 amino acid residues of the sequence set forth in SEQ ID NO:8, provided that amino acids 20, 30, 31, 48, 63, 74, 95, 127, 173, 239, 336, 366, 491, 515, 564, 585, 629, 640, 669, 676, 679, 749 and 750 of the sequence set forth in SEQ ID NO:8 are conserved; e) a protein comprising an amino acid sequence set forth in SEQ ID NO:10; 11 Aug 2025 f) a protein comprising: (i) an amino acid sequence set forth in SEQ ID NO:12; an amino acid sequence that is at least 95% identical to the sequence set forth in SEQ ID NO:12; or (iii) an immunogenic fragment of (i), wherein the immunogenic fragment comprises amino acids corresponding to at least 349 amino acid residues of the sequence set forth in SEQ ID NO:12; and g) a protein comprising: (i) an amino acid sequence set forth in SEQ ID NO:14; 2023200517 an amino acid sequence that is at least 95% identical to the sequence set forth in SEQ ID NO:14; or (iii) an immunogenic fragment of (i), wherein the immunogenic fragment comprises amino acids corresponding to at least 129 amino acid residues of the sequence set forth in SEQ ID NO:14.

11. The composition of claim 10, comprising at least one prostate cancer antigen selected from a) or b) and at least one prostate cancer antigen selected from the group consisting of elements c), d), e), f), and g).

12. The composition of claim 9 or 10, comprising at least one prostate cancer antigen selected from the group of proteins consisting of: a protein comprising the amino acid sequence set forth in SEQ ID NO:2; a protein comprising the amino acid sequence set forth in SEQ ID NO:4; a protein comprising the amino acid sequence set forth in SEQ ID NO:6; a protein comprising the amino acid sequence set forth in SEQ ID NO:8; a protein comprising the amino acid sequence set forth in SEQ ID NO:10; a protein comprising the amino acid sequence set forth in SEQ ID NO:12; and a protein comprising the amino acid sequence set forth in SEQ ID NO:14.

13. The composition of any one of claims 10-12, further comprising a pharmaceutically acceptable excipient.

14. A method of treating an individual who has been diagnosed with prostate cancer, comprising delivering to said individual the composition of any one of claims 10-13.

15. Use of the composition of any one of claims 10-14 in the manufacture of a medicament for treating prostate cancer in an individual who has been diagnosed therewith.