WO2024243542A1

WO2024243542A1 - Compositions containing p53 peptide amphiphiles and methods of use thereof

Info

Publication number: WO2024243542A1
Application number: PCT/US2024/031064
Authority: WO
Inventors: Peter C. DEMUTH; Christopher M. HAQQ; Martin P. STEINBUCK
Original assignee: Elicio Therapeutics, Inc.
Priority date: 2023-05-25
Filing date: 2024-05-24
Publication date: 2024-11-28

Abstract

Disclosed herein are compounds including albumin-binding domain and a mutant or wild-type p53 peptide, as well as pharmaceutically acceptable salts thereof. Furthermore, disclosed herein are methods for inducing an immune response in a subject, and methods of administering such compounds to induce an immune response in a subject.

Description

COMPOSITIONS CONTAINING P53 PEPTIDE AMPHIPHILES AND METHODS OF USE THEREOF

SEQUENCE LISTING

The instant application contains a Sequence Listing which has been submitted electronically in XML format and is hereby incorporated by reference in its entirety. Said XML copy, created May 23, 2024, is named 51026-059WO3_Sequence_Listing_5_23_24.xml and is 127,781 bytes in size.

BACKGROUND OF THE INVENTION

Vaccines are used to stimulate an immune response in an individual to provide protection against and/or treatment for a particular disease. Some vaccines include an antigen to induce an immune response. Immune responses as a result of vaccination have made an enormous contribution to both human and animal health. Since the invention of the first vaccine in 1796, vaccines have come to be considered the most successful method for preventing many infectious diseases by provoking an immune response in a subject. According to the World Health Organization, immunization currently prevents 2-3 million deaths every year across all age groups. The purpose of vaccination is to generate a strong and lasting immune response providing long-term protection against infection. However, many vaccines do not currently induce optimal immunity.

There remains a need to develop new and improved compositions and methods for inducing immune responses in a subject thereof.

SUMMARY OF THE INVENTION

In an aspect, the disclosure provides a compound comprising an albumin-binding domain and a p53 peptide or a mutant p53 peptide, or a pharmaceutically acceptable salt thereof. In some embodiments, the p53 peptide 5 to 50 (e.g., 5, 6, 7, 8, 9, 11 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 35, 36, 37, 38, 39, 40, 41 , 42, 43, 44, 45, 46, 47, 48, 49, or 50) amino acid fragment of SEQ ID NO: 1 . In some embodiments, the p53 peptide is a 20 to 40 (e.g., 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 35, 36, 37, 38, 39, or 40) amino acid fragment of SEQ ID NO: 1 . In some embodiments, the p53 peptide is a 20 to 30 (e.g., 21 , 22, 23, 24, 25, 26, 27, 28, 29, or 30) amino acid fragment of SEQ ID NO: 1 . In some embodiments, the p53 peptide is a 30 amino acid fragment of SEQ ID NO: 1 .In some embodiments, the mutant p53 peptide comprises at least one amino acid substitution relative to the corresponding sequence of SEQ ID NO: 1 . In some embodiments, the mutant p53 peptide 5 to 50 (e.g., 5, 6, 7, 8, 9, 11 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 35, 36, 37, 38, 39, 40, 41 , 42, 43, 44, 45, 46, 47, 48, 49, or 50) amino acid fragment of SEQ ID NO: 1 comprising at least one amino acid substitution relative to the corresponding sequence of SEQ ID NO: 1 . In some embodiments, the mutant p53 peptide is a 20 to 40 (e.g., 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 35, 36, 37, 38, 39, or 40) amino acid fragment of SEQ ID NO: 1 comprising at least one amino acid substitution relative to the corresponding sequence of SEQ ID NO: 1 . In some embodiments, the mutant p53 peptide is a 20 to 30 (e.g., 21 , 22, 23, 24, 25, 26, 27, 28, 29, or 30) amino acid fragment of SEQ ID NO: 1 comprising at least one amino acid substitution relative to the corresponding sequence of SEQ ID NO: 1 . In some embodiments, the mutant p53 peptide is a 30 amino acid fragment of SEQ ID NO: 1 comprising at least one amino acid substitution relative to the corresponding sequence of SEQ ID NO: 1 . In some embodiments, the mutant p53 peptide comprises at least one of the amino acid substitutions described in TABLE 1 .

In some embodiments, the mutant p53 peptide comprises a fragment of SEQ ID NO: 1 , wherein the fragment comprises an amino acid substitution at the Cys occupying amino acid position 135 from the N-terminus of SEQ ID NO: 1 . In some embodiments, the amino acid substitution at the Cys occupying position 135 of SEQ ID NO: 1 is a C135Y substitution. In some embodiments, the mutant p53 peptide comprises a fragment of SEQ ID NO: 1 , wherein the fragment comprises an amino acid substitution at the Cys occupying amino acid position 141 from the N-terminus of SEQ ID NO: 1 . In some embodiments, the amino acid substitution at the Cys occupying position 141 of SEQ ID NO: 1 is a C141 Y substitution. In some embodiments, the mutant p53 peptide comprises a fragment of SEQ ID NO: 1 , wherein the fragment comprises an amino acid substitution at the Pro occupying amino acid position 151 from the N- terminus of SEQ ID NO: 1 . In some embodiments, the amino acid substitution at the Pro occupying position 151 of SEQ ID NO: 1 is a P151 S substitution. In some embodiments, the mutant p53 peptide comprises a fragment of SEQ ID NO: 1 , wherein the fragment comprises an amino acid substitution at the Pro occupying amino acid position 152 from the N-terminus of SEQ ID NO: 1 . In some embodiments, the amino acid substitution at the Pro occupying position 152 of SEQ ID NO: 1 is a P152L substitution. In some embodiments, the mutant p53 peptide comprises a fragment of SEQ ID NO: 1 , wherein the fragment comprises an amino acid substitution at the Vai occupying amino acid position 157 from the N- terminus of SEQ ID NO: 1 . In some embodiments, the amino acid substitution at the Vai occupying position 157 of SEQ ID NO: 1 is a V157F substitution. In some embodiments, the mutant p53 peptide comprises a fragment of SEQ ID NO: 1 , wherein the fragment comprises an amino acid substitution at the Arg occupying amino acid position 158 from the N-terminus of SEQ ID NO: 1 . In some embodiments, the amino acid substitution at the Arg occupying position 158 of SEQ ID NO: 1 is a R158H substitution. In some embodiments, the amino acid substitution at the Arg occupying position 158 of SEQ ID NO: 1 is a R158L substitution. In some embodiments, the mutant p53 peptide comprises a fragment of SEQ ID NO: 1 , wherein the fragment comprises an amino acid substitution at the Ala occupying amino acid position 161 from the N-terminus of SEQ ID NO: 1 . In some embodiments, the amino acid substitution at the Ala occupying position 161 of SEQ ID NO: 1 is a A161T substitution. In some embodiments, the mutant p53 peptide comprises a fragment of SEQ ID NO: 1 , wherein the fragment comprises an amino acid substitution at the Tyr occupying amino acid position 163 from the N-terminus of SEQ ID NO: 1 . In some embodiments, the amino acid substitution at the Tyr occupying position 163 of SEQ ID NO: 1 is a Y163C substitution. In some embodiments, the mutant p53 peptide comprises a fragment of SEQ ID NO: 1 , wherein the fragment comprises an amino acid substitution at the Vai occupying amino acid position 173 from the N-terminus of SEQ ID NO: 1 . In some embodiments, the amino acid substitution at the Vai occupying position 173 of SEQ ID NO: 1 is a V173M substitution. In some embodiments, the amino acid substitution at the Vai occupying position 173 of SEQ ID NO: 1 is a V173L substitution. In some embodiments, the mutant p53 peptide comprises a fragment of SEQ ID NO: 1 , wherein the fragment comprises an amino acid substitution at the Arg occupying amino acid position 175 from the N-terminus of SEQ ID NO: 1 . In some embodiments, the amino acid substitution at the Arg occupying position 175 of SEQ ID NO: 1 is a R175H substitution. In some embodiments, the mutant p53 peptide comprises a fragment of SEQ ID NO: 1 , wherein the fragment comprises an amino acid substitution at the Cys occupying amino acid position 176 from the N-terminus of SEQ ID NO: 1 . In some embodiments, the amino acid substitution at the Cys occupying position 176 of SEQ ID NO: 1 is a C176F substitution. In some embodiments, the amino acid substitution at the Cys occupying position 176 of SEQ ID NO: 1 is a C176Y substitution. In some embodiments, the mutant p53 peptide comprises a fragment of SEQ ID NO: 1 , wherein the fragment comprises an amino acid substitution at the His occupying amino acid position 179 from the N-terminus of SEQ ID NO: 1 . In some embodiments, the amino acid substitution at the His occupying position 179 of SEQ ID NO: 1 is a H179Y substitution. In some embodiments, the amino acid substitution at the His occupying position 179 of SEQ ID NO: 1 is a H179R substitution. In some embodiments, the mutant p53 peptide comprises a fragment of SEQ ID NO: 1 , wherein the fragment comprises an amino acid substitution at the His occupying amino acid position 193 from the N-terminus of SEQ ID NO: 1 . In some embodiments, the amino acid substitution at the His occupying position 193 of SEQ ID NO: 1 is a H193R substitution. In some embodiments, the mutant p53 peptide comprises a fragment of SEQ ID NO: 1 , wherein the fragment comprises an amino acid substitution at the lie occupying amino acid position 195 from the N-terminus of SEQ ID NO: 1 . In some embodiments, the amino acid substitution at the lie occupying position 195 of SEQ ID NO: 1 is a 1195T substitution. In some embodiments, the mutant p53 peptide comprises a fragment of SEQ ID NO: 1 , wherein the fragment comprises an amino acid substitution at the Tyr occupying amino acid position 205 from the N- terminus of SEQ ID NO: 1 . In some embodiments, the amino acid substitution at the Tyr occupying position 205 of SEQ ID NO: 1 is a Y205C substitution. In some embodiments, the mutant p53 peptide comprises a fragment of SEQ ID NO: 1 , wherein the fragment comprises an amino acid substitution at the His occupying amino acid position 214 from the N-terminus of SEQ ID NO: 1 . In some embodiments, the amino acid substitution at the His occupying position 214 of SEQ ID NO: 1 is a H214R substitution. In some embodiments, the mutant p53 peptide comprises a fragment of SEQ ID NO: 1 , wherein the fragment comprises an amino acid substitution at the Vai occupying amino acid position 216 from the N- terminus of SEQ ID NO: 1 . In some embodiments, the amino acid substitution at the lie occupying position 216 of SEQ ID NO: 1 is a V216M substitution. In some embodiments, the mutant p53 peptide comprises a fragment of SEQ ID NO: 1 , wherein the fragment comprises an amino acid substitution at the Tyr occupying amino acid position 220 from the N-terminus of SEQ ID NO: 1 . In some embodiments, the amino acid substitution at the Tyr occupying position 220 of SEQ ID NO: 1 is a Y220C substitution. In some embodiments, the mutant p53 peptide comprises a fragment of SEQ ID NO: 1 , wherein the fragment comprises an amino acid substitution at the Tyr occupying amino acid position 234 from the N- terminus of SEQ ID NO: 1 . In some embodiments, the amino acid substitution at the Tyr occupying position 234 of SEQ ID NO: 1 is a Y234C substitution.

In some embodiments, the mutant p53 peptide comprises a fragment of SEQ ID NO: 1 , wherein the fragment comprises an amino acid substitution at the Tyr occupying amino acid position 236 from the N-terminus of SEQ ID NO: 1 . In some embodiments, the amino acid substitution at the Tyr occupying position 236 of SEQ ID NO: 1 is a Y236C substitution. In some embodiments, the mutant p53 peptide comprises a fragment of SEQ ID NO: 1 , wherein the fragment comprises an amino acid substitution at the Met occupying amino acid position 237 from the N-terminus of SEQ ID NO: 1 . In some embodiments, the amino acid substitution at the Met occupying position 237 of SEQ ID NO: 1 is a M237I substitution. In some embodiments, the mutant p53 peptide comprises a fragment of SEQ ID NO: 1 , wherein the fragment comprises an amino acid substitution at the Cys occupying amino acid position 238 from the N- terminus of SEQ ID NO: 1 . In some embodiments, the amino acid substitution at the Cys occupying position 238 of SEQ ID NO: 1 is a C238Y substitution. In some embodiments, the mutant p53 peptide comprises a fragment of SEQ ID NO: 1 , wherein the fragment comprises an amino acid substitution at the Ser occupying amino acid position 241 from the N-terminus of SEQ ID NO: 1 . In some embodiments, the amino acid substitution at the Ser occupying position 241 of SEQ ID NO: 1 is a S241 F substitution. In some embodiments, the mutant p53 peptide comprises a fragment of SEQ ID NO: 1 , wherein the fragment comprises an amino acid substitution at the Cys occupying amino acid position 242 from the N- terminus of SEQ ID NO: 1 . In some embodiments, the amino acid substitution at the Cys occupying position 242 of SEQ ID NO: 1 is a C242F substitution. In some embodiments, the mutant p53 peptide comprises a fragment of SEQ ID NO: 1 , wherein the fragment comprises an amino acid substitution at the Gly occupying amino acid position 245 from the N-terminus of SEQ ID NO: 1 . In some embodiments, the amino acid substitution at the Gly occupying position 245 of SEQ ID NO: 1 is a G245C substitution. In some embodiments, the amino acid substitution at the Gly occupying position 245 of SEQ ID NO: 1 is a G245D substitution. In some embodiments, the amino acid substitution at the Gly occupying position 245 of SEQ ID NO: 1 is a G245S substitution. In some embodiments, the amino acid substitution at the Gly occupying position 245 of SEQ ID NO: 1 is a G245V substitution. In some embodiments, the mutant p53 peptide comprises a fragment of SEQ ID NO: 1 , wherein the fragment comprises an amino acid substitution at the Arg occupying amino acid position 248 from the N-terminus of SEQ ID NO: 1 . In some embodiments, the amino acid substitution at the Arg occupying position 248 of SEQ ID NO: 1 is a R248L substitution. In some embodiments, the amino acid substitution at the Arg occupying position 248 of SEQ ID NO: 1 is a R248Q substitution. In some embodiments, the amino acid substitution at the Arg occupying position 248 of SEQ ID NO: 1 is a R248W substitution. In some embodiments, the mutant p53 peptide comprises a fragment of SEQ ID NO: 1 , wherein the fragment comprises an amino acid substitution at the Arg occupying amino acid position 249 from the N-terminus of SEQ ID NO: 1 . In some embodiments, the amino acid substitution at the Arg occupying position 249 of SEQ ID NO: 1 is a R249S substitution. In some embodiments, the mutant p53 peptide comprises a fragment of SEQ ID NO: 1 , wherein the fragment comprises an amino acid substitution at the Gly occupying amino acid position 266 from the N-terminus of SEQ ID NO: 1 . In some embodiments, the amino acid substitution at the Gly occupying position 266 of SEQ ID NO: 1 is a G266E substitution. In some embodiments, the amino acid substitution at the Gly occupying position 266 of SEQ ID NO: 1 is a G266R substitution. In some embodiments, the mutant p53 peptide comprises a fragment of SEQ ID NO: 1 , wherein the fragment comprises an amino acid substitution at the Vai occupying amino acid position 272 from the N-terminus of SEQ ID NO: 1 . In some embodiments, the amino acid substitution at the Vai occupying position 272 of SEQ ID NO: 1 is a V272M substitution. In some embodiments, the mutant p53 peptide comprises a fragment of SEQ ID NO: 1 , wherein the fragment comprises an amino acid substitution at the Arg occupying amino acid position 273 from the N-terminus of SEQ ID NO: 1 . In some embodiments, the amino acid substitution at the Arg occupying position 273 of SEQ ID NO: 1 is a R273H substitution. In some embodiments, the amino acid substitution at the Arg occupying position 273 of SEQ ID NO: 1 is a R273C substitution. In some embodiments, the amino acid substitution at the Arg occupying position 273 of SEQ ID NO: 1 is a R273L substitution. In some embodiments, the mutant p53 peptide comprises a fragment of SEQ ID NO: 1 , wherein the fragment comprises an amino acid substitution at the Cys occupying amino acid position 275 from the N-terminus of SEQ ID NO: 1 . In some embodiments, the amino acid substitution at the Cys occupying position 275 of SEQ ID NO: 1 is a C275Y substitution. In some embodiments, the mutant p53 peptide comprises a fragment of SEQ ID NO: 1 , wherein the fragment comprises an amino acid substitution at the Pro occupying amino acid position 278 from the N- terminus of SEQ ID NO: 1 . In some embodiments, the amino acid substitution at the Pro occupying position 278 of SEQ ID NO: 1 is a P278S substitution. In some embodiments, the amino acid substitution at the Pro occupying position 278 of SEQ ID NO: 1 is a P278L substitution. In some embodiments, the mutant p53 peptide comprises a fragment of SEQ ID NO: 1 , wherein the fragment comprises an amino acid substitution at the Arg occupying amino acid position 280 from the N-terminus of SEQ ID NO: 1 . In some embodiments, the amino acid substitution at the Arg occupying position 280 of SEQ ID NO: 1 is a R280T substitution. In some embodiments, the amino acid substitution at the Arg occupying position 280 of SEQ ID NO: 1 is a R280K substitution. In some embodiments, the mutant p53 peptide comprises a fragment of SEQ ID NO: 1 , wherein the fragment comprises an amino acid substitution at the Arg occupying amino acid position 282 from the N-terminus of SEQ ID NO: 1 . In some embodiments, the amino acid substitution at the Arg occupying position 282 of SEQ ID NO: 1 is a R282W substitution. In some embodiments, the mutant p53 peptide comprises a fragment of SEQ ID NO: 1 , wherein the fragment comprises an amino acid substitution at the Glu occupying amino acid position 285 from the N- terminus of SEQ ID NO: 1 . In some embodiments, the amino acid substitution at the Glu occupying position 285 of SEQ ID NO: 1 is a E285K substitution. In some embodiments, the mutant p53 peptide comprises a fragment of SEQ ID NO: 1 , wherein the fragment comprises an amino acid substitution at the Glu occupying amino acid position 286 from the N-terminus of SEQ ID NO: 1 . In some embodiments, the amino acid substitution at the Glu occupying position 286 of SEQ ID NO: 1 is a E286K substitution.

In some embodiments, the mutant p53 peptide consists of the amino acid sequence of YMCNSSCMGGMNWRPILTIITLEDS (SEQ ID NO: 2), or a fragment thereof. In some embodiments, the mutant p53 peptide consists of the amino acid sequence of YMCNSSCMGGMNQRPILTIITLEDS (SEQ ID NO:3), or a fragment thereof. In some embodiments, the mutant p53 peptide comprises a 10 amino acid fragment of SEQ ID NO: 2 or 3.

In some embodiments, the mutant p53 peptide comprises the amino acid sequence NWRPILTIIT (SEQ ID NO: 46) or NQRPILTIIT (SEQ ID NO: 47). In some embodiments, the mutant p53 peptide comprises a 10 or 25 (e.g., 10, 11 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, or 25) amino acid fragment of SEQ ID NO: 2 or 3. In some embodiments, the mutant p53 peptide comprises the amino acid sequence NSSCMGGMNWRPILTIIT (SEQ ID NO: 48) or NSSCMGGMNQRPILTIIT (SEQ ID NO: 49).

In some embodiments, the mutant p53 peptide consists of the amino acid sequence of DSSGNLLGRNSFEVHVCACPGRDRRTEEEN (SEQ ID NO: 99), or a fragment thereof.

In some embodiments, the mutant p53 peptide consists of the amino acid sequence of DSSGNLLGRNSFEVCVCACPGRDRRTEEEN (SEQ ID NO: 113), or a fragment thereof.

In some embodiments, the mutant p53 peptide consists of the amino acid sequence of NSFEVCVCACPGRDWRTEEENLRKKGEPHH (SEQ ID NO: 114), or a fragment thereof.

In some embodiments, the mutant p53 peptide consists of the amino acid sequence of TIHYNYMCNSSCMGSMNRRPILTIITLEDS (SEQ ID NO: 115), or a fragment thereof. In some embodiments, the mutant p53 peptide consists of the amino acid sequence of NYMCNSSCMGGMNRSPILTIITLEDSSGNL (SEQ ID NO: 116), or a fragment thereof.

In some embodiments, the mutant p53 peptide consists of the amino acid sequence of LDDRNTFRHSVVVPCEPPEVGSDCTTIHYN (SEQ ID NO: 117), or a fragment thereof.

In some embodiments, the mutant p53 peptide consists of the amino acid sequence of RLGFLHSGTAKSVTCTYSPALNKMFYQLAK (SEQ ID NO: 118), or a fragment thereof.

In some embodiments, the mutant p53 peptide consists of the amino acid sequence of PVQLWVDSTPPPGTRVHAMAIYKQSQHMTE (SEQ ID NO: 119), or a fragment thereof.

In some embodiments, the mutant p53 peptide consists of the amino acid sequence of LRVEYLDDRNTFRRSVVVPYEPPEVGSDCT (SEQ ID NO: 120), or a fragment thereof.

In some embodiments, the mutant p53 peptide consists of the amino acid sequence of RAMAIYKQSQHMTEVVRHCPHHERCSDSDG (SEQ ID NO: 122), or a fragment thereof.

In another aspect, the disclosure provides a compound comprising an albumin-binding domain and a p53 peptide, or a pharmaceutically acceptable salt thereof, where the p53 peptide comprises the sequence MAIYKQSQHMTEVVRRCPHHERCSDSDGLAP (SEQ ID NO:100), or a fragment thereof. In one embodiment, the fragment comprises the sequence AIYKQSQHM (SEQ ID NO:101 ).

In another aspect, the disclosure provides a compound comprising an albumin-binding domain and a p53 peptide, or a pharmaceutically acceptable salt thereof, where the p53 peptide comprises the sequence EGNLRVEYLDDRNTFRHSVVVPCEPPEVGSD (SEQ ID NO:102), or a fragment thereof. IN one embodiment, the fragment comprises the sequence EYLDDRNTF (SEQ ID NO:103).

In another aspect, the disclosure provides a compound comprising an albumin-binding domain and a p53 peptide, or a pharmaceutically acceptable salt thereof, where the p53 peptide comprises the sequence YLDDRNTFRHSVVVPYEPPEVGSDCTTIHYN (SEQ ID NO:104) or a fragment thereof. In one embodiment, the fragment comprises the sequence VVPYEPPEV (SEQ ID NO:105.

In another aspect, the disclosure provides a compound comprising an albumin-binding domain and a p53 peptide, or a pharmaceutically acceptable salt thereof, where the p53 peptide comprises the sequence TTIHYNYMCNSSCMGGMNRRPILTIITLEDSSGNL (SEQ ID NO:106) or a fragment thereof.

In another aspect, the disclosure provides a compound comprising an albumin-binding domain and a p53 peptide, or a pharmaceutically acceptable salt thereof, where the p53 peptide comprises the sequence EDSSGNLLGRNSFEVRVCACPGRDRRTEEENLRKK (SEQ ID NQ:107), or a fragment thereof. In one embodiment, the fragment comprises the sequence LLGRNSFEV (SEQ ID NO:108.

In another aspect, the disclosure provides a compound comprising an albumin-binding domain and a p53 peptide, or a pharmaceutically acceptable salt thereof, where the p53 peptide comprises the sequence RLGFLHSGTAKSVTC (SEQ ID NO:109), or a fragment thereof.

In another aspect, the disclosure provides a compound comprising an albumin-binding domain and a p53 peptide, or a pharmaceutically acceptable salt thereof, where the p53 peptide comprises the sequence STPPPGTRV (SEQ ID NO:110), or a fragment thereof.

In another aspect, the disclosure provides a compound comprising an albumin-binding domain and a p53 peptide, or a pharmaceutically acceptable salt thereof, where the p53 peptide comprises the sequence TYPALNKMF (SEQ ID NO:111 ), or a fragment thereof. In another aspect, the disclosure provides a compound comprising an albumin-binding domain and a p53 peptide, or a pharmaceutically acceptable salt thereof, where the p53 peptide comprises the sequence RMPEAAPPV (SEQ ID NO:112), or a fragment thereof.

In another aspect, the disclosure provides a compound comprising an albumin-binding domain and a p53 peptide, or a pharmaceutically acceptable salt thereof, where the p53 peptide comprises the sequence TEDPGPDEAPRMPEAAPPVAPAPAAPTPAA (SEQ ID NO:121 ), or a fragment thereof.

In some embodiments, the peptide optionally includes an N-terminal modification. In some embodiments, the peptide includes an N-terminal modification. In some embodiments, the N-terminal modification is the addition of an acetylcysteine. In some embodiments, the N-terminal modification is the addition of a des-aminocysteine homolog. In some embodiments, the des-aminocysteine homolog is 3- mercaptopropionic acid or mercaptoacetic acid. In some embodiments, the N-terminus of the peptide is bonded or linked to the albumin-binding domain. In some embodiments, the C-terminus of the peptide is bonded or linked to the albumin-binding domain.

In some embodiments, the albumin-binding domain includes a lipid. In some embodiments, the lipid is a diacyl lipid. In some embodiments, the diacyl lipid comprises acyl chains comprising 12-30 hydrocarbon units, 14-25 hydrocarbon units, 16-20 hydrocarbon units, or 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, or 30 hydrocarbon units. In some embodiments, the lipid is 1 ,2- distearoyl-sn-glycero-3-phosphoethanolamine (DSPE). In some embodiments, the peptide is bonded or linked by a linker to the following lipid:

or a salt thereof, wherein X is O or S. In some embodiments, the linker is selected from the group consisting of a hydrophilic polymer, a string of hydrophilic amino acids, a polysaccharide, and an oligonucleotide, or a combination thereof. In some embodiments, the linker comprises "N" polyethylene glycol units, wherein N is between 24-50. In some embodiments, the linker comprises PEG24-amido-PEG24.

In another aspect, the disclosure provides a method of inducing an immune response in a subject including administering any one of the compounds described herein or pharmaceutically acceptable salt thereof to the subject. In some embodiments, the method further includes administering an adjuvant to the subject. In some embodiments, the compound or pharmaceutically acceptable salt thereof is administered subcutaneously, intramuscularly, intravenously, or transmucosally. In some embodiments, the subject is a mammal. In some embodiments, the subject is a human.

In another aspect, the disclosure provides a compound or pharmaceutically acceptable salt thereof comprising an albumin-binding domain and a mutant p53 peptide for use in a method of inducing an immune response in a subject, wherein the method comprises administering any one of the compounds described herein to the subject. In some embodiments, the method further includes administering an adjuvant to the subject. In some embodiments, the compound or pharmaceutically acceptable salt thereof is administered subcutaneously, intramuscularly, intravenously, or transmucosally. In some embodiments, the subject is a mammal. In some embodiments, the subject is a human.

In another aspect, the disclosure provides a pharmaceutical composition comprising any one of the compounds described herein or pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

In another aspect, the disclosure provides a kit comprising any one of the compounds described herein or pharmaceutically acceptable salt thereof or the pharmaceutical composition the described herein and instructions for administration.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 A is a drawing of a single-stranded amphiphile (AMP) conjugated to a PEG-48 linker which is conjugated to a mutant p53 peptide, which is a 10,18, or 30 amino acid fragment of the amino acid sequence of SEQ ID NO: 2. A 10,18, or 30 amino acid fragment of the amino acid sequence of SEQ ID NO: 126 is also shown.

FIG. 1B is a table of mutant p53 peptides including R248W, R248Q, R175H, R273H, R273C, R282W, G245S, R249S, Y220C, C135Y, R158H, and H214R and wildtype as shown in SEQ ID NOs: 99, 115-122, and 127-129.

FIG. 2A and FIG. 2B are graphs showing the splenocyte IFNy ELISpot responses of mice that were administered a vaccine including a soluble or an amphiphilic peptide as shown in FIG. 1 after two doses (FIG. 2A) and three doses (FIG. 2B).

FIG. 3A and FIG. 3B are graphs showing the splenocyte IFNy ELISpot responses of mice that were administered a vaccine including a soluble or an amphiphilic mutant p53 peptide after two doses (FIG. 3A) and three doses (FIG. 3B), wherein the mutant p53 peptide is a 10 or 18 amino acid fragment of the amino acid sequence of SEQ ID NO: 3.

FIG. 4 is a graph showing the splenocyte IFNy ELISpot responses of C57BI6 mice that were administered a vaccine of a soluble (SOL) or amphiphile (AMP) p53 R248W 18-mer peptide at a concentration of 1 .25 nmol, 5 nmol, 10 nmol or 20 nmol.

FIG. 5A-FIG. 5E are graphs showing the concentration of Granzyme B (FIG. 5A), INFy (FIG. 5B), TNFa (FIG. 5C), GM-CSF (FIG. 5D), and IL2 (FIG. 5E) from splenocytes 7 days after dose 3 for C57BL/6J mice that were administered a vaccine a soluble (SOL) or amphiphile (AMP) p53 R248W 18- mer peptide at a concentration of 1 .25 nmol, 5 nmol, 10 nmol or 20 nmol.

FIG. 6A and FIG. 6B are graphs showing the percentage of cytokines, including (from top to bottom in each column) IFNy and TNFa, only IFNy, and only TNFa in CD4+ cells (FIG. 6A) and CD8+ cells (FIG. 6B) found in peripheral blood cells collected C57BL/6J mice that were administered a vaccine a soluble (SOL) or amphiphile (AMP) p53 R248W 18-mer peptide at a concentration of 1 .25 nmol, 5 nmol, 10 nmol or 20 nmol.

FIG. 7 is a graph showing the splenocyte IFNy ELISpot responses of C57BI6 mice that were administered a vaccine of a soluble (SOL) or amphiphile (AMP) p53 R248W 18-mer peptide at a concentration of 1 .25 nmol and an adjuvant at a concentration of 5 nmol on a dosing schedule of three doses administered weekly, four doses administered weekly, three doses administered weekly with a skipped week on the third consecutive week, three doses administered biweekly (AMP), or five doses administered weekly (SOL).

FIG. 8A-FIG. 8E are graphs showing the concentration of Granzyme B (FIG. 8A), INFy (FIG. 8B), TNFa (FIG. 8C), GM-CSF (FIG. 8D), and IL2 (FIG. 8E) from splenocytes 7 days after dose 3 for C57BI6 mice that were administered a vaccine of a soluble (SOL) or amphiphile (AMP) p53 R248W 18-mer peptide at a concentration of 1 .25 nmol and an adjuvant at a concentration of 5 nmol on a dosing schedule of three doses administered weekly, four doses administered weekly, three doses administered weekly with a skipped week on the third consecutive week, three doses administered biweekly (AMP), or five doses administered weekly (SOL).

FIG. 9 is a graph showing the percentage of cytokines, including (from top to bottom in each column) IFNy and TNFa, only TNFa, and only IFNy, in CD4+ cells found in peripheral blood cells collected C57BL/6J mice that were administered a vaccine of a soluble (SOL) or amphiphile (AMP) p53 R248W 18-mer peptide at a concentration of 1 .25 nmol and an adjuvant at a concentration of 5 nmol on a dosing schedule of three doses administered weekly, four doses administered weekly, three doses administered weekly with a skipped week on the third consecutive week, three doses administered bi weekly (AMP), or five doses administered weekly (SOL).

FIG. 10A and FIG. 10B are graphs showing the percentage of target cell killing that occurred after administration of p53 R248W 18-mer peptide at a concentration of 1 .25 nmol and an adjuvant at a concentration of 5 nmol that were dosed five times weekly and were analyzed post-dose 3 (PD3) and post-dose 5 (PD5).

FIG. 11A-FIG. 11D are graphs showing the splenocyte post-dose 2 (PD2) (FIG. 11 A), post-dose 3 (PD3) (FIG. 11 B), wild-type control (FIG. 11 C) and lung-resident lymphocytes (FIG. 11 D) IFNy ELISpot responses of C57BI6 mice that were administered a vaccine of a soluble (SOL) or amphiphile (AMP) p53 R248W 10-mer, 18-mer, or 30-mer peptide at a concentration of 1 .25 nmol and an adjuvant at a concentration of 5 nmol administered on days 7, 14 and 28.

FIG. 12A and FIG. 12B are graphs showing the amount of various cytokines, including (from top to bottom in each column) IFNy+TNFa+IL2, IFNy and IL2, IFNy and TNFa, IL2, TNFa, and IFNy, found in splenocytes and lung-resident lymphocytes of C57BI6 mice that were administered a vaccine of a soluble (SOL) or amphiphile (AMP) p53 R248W 10-mer, 18-mer, or 30-mer peptide at a concentration of 1 .25 nmol and an adjuvant at a concentration of 5 nmol administered on days 7, 14 and 28.

FIG. 13 is a graph showing the splenocyte IFNy ELISpot analysis for Granzyme B post-dose 3 (PD3) of C57BI6 mice that were administered a vaccine of a soluble (SOL) or amphiphile (AMP) p53 R248W 10-mer, 18-mer, or 30-mer peptide at a concentration of 1 .25 nmol and an adjuvant at a concentration of 5 nmol administered on days 7, 14 and 28.

FIG. 14A and FIG. 14B are graphs showing the percentage of cytokines, including (from top to bottom in each column) IFNy and TNFa, only IFNy, and only TNFa in CD4+ cells (FIG. 14A) and CD8+ - cells (FIG. 14B) found in peripheral blood cells collected from C57BL/6J mice that were administered a vaccine a soluble (SOL) or amphiphile (AMP) p53 R248W 10-mer, 18-mer, or 30-mer at a concentration of 1 .25 nmol and an adjuvant at a concentration of 5 nmol.

FIG. 15A-FIG 15F are graphs showing the percentage of cytokines, including (from top to bottom in each column) IFNy and TNFa, only IFNy, and only TNFa found in CD4+ and CD8+ cells isolated from the spleen (FIG. 15A and FIG. 15D), CD4+ and CD8+ cells isolated lung (FIG. 15B and FIG. 15E), and CD4+ and CD8+ cells isolated peripheral blood cells (FIG. 15C and FIG. 15F) collected from C57BL/6J mice that were administered a vaccine a soluble (SOL) or amphiphile (AMP) p53 R248W 10-mer, 18-mer, or 30-mer at a concentration of 1 .25 nmol and an adjuvant at a concentration of 5 nmol .

FIG. 16A-FIG. 16E are graphs showing the concentration of Granzyme B (FIG. 16A), INFy (FIG. 16B), TNFa (FIG. 16C), GM-CSF (FIG. 16D), and IL2 (FIG. 16E) from splenocytes 7 days after dose 3 for C57BL/6J mice that were administered a vaccine a soluble (SOL) or amphiphile (AMP) p53 R248W 10- mer, 18-mer, or 30-mer at a concentration of 1 .25 nmol and an adjuvant at a concentration of 5 nmol .

FIG. 17 is a graph showing the splenocyte IFNy ELISpot response of post-dose 5 (PD5) of C57BI6 mice that were administered a vaccine of a soluble (SOL) or amphiphile (AMP) p53 R248W 18- mer peptide at a concentration of 1 .25 nmol and an adjuvant at a concentration of 5 nmol administered weekly.

FIG. 18A-FIG. 18E are graphs showing the concentration of Granzyme B (FIG. 18A), INFy (FIG. 18B), TNFa (FIG. 18C), GM-CSF (FIG. 18D), and IL2 (FIG. 18E) from splenocytes 7 days after dose 5 for C57BI6 mice that were administered a vaccine of a soluble (SOL) or amphiphile (AMP) p53 R248W 18- mer peptide at a concentration of 1 .25 nmol and an adjuvant at a concentration of 5 nmol administered weekly.

FIG. 19 is a series of graphs showing the splenocyte IFNy ELISpot response of post-dose 3 (PD3) of C57BI6 mice that were administered a vaccine of soluble (SOL) or amphiphile (AMP) p53 R248W, R175H, R273H, G245S, Y220C, C135Y, R158H, and H214R 30-mer peptides at a concentration of 1 .25 nmol and an adjuvant at a concentration of 5 nmol administered bi-weekly.

FIG. 20A-FIG. 20C are graphs showing the splenocyte post-dose 2 (PD2) (FIG. 20A) and postdose 3 (PD3) (FIG.20B) IFNy ELISpot responses of C57BI6 mice that were administered a vaccine of a SOL or AMP p53 R248Q 30-mer peptide at a concentration of 1 .25 nmol and an adjuvant at a concentration of 5 nmol, and PD3 mice that were administered a vaccine of an AMP p53 R248Q 30-mer peptide at a concentration of 5 nmol and an adjuvant at a concentration of 10 nmol (FIG. 20C).

FIG. 21A-FIG. 21C are graphs showing the splenocyte IFNy ELISpot analysis for Granzyme B (FIG. 21 A), cytokine FluoroSpot analysis (FIG. 21 B), and the percentage of cytokines, including (from top to bottom in each column) IFNy and TNFa, only IFNy, and only TNFa in CD8+ cells (FIG. 21 C) post-dose 3 (PD3) of C57BI6 mice that were administered a vaccine of amphiphile (AMP) p53 R248Q 30-mer peptide at a concentration of 5 nmol and an adjuvant at a concentration of 10 nmol.

FIG. 22A-FIG. 22E are graphs showing the concentration of Granzyme B (FIG. 22A), IFNy (FIG. 22B), TNFa (FIG. 22C), GM-CSF (FIG. 22D), and IL2 (FIG. 22E) from splenocytes 7 days after dose 3 for C57BI6 mice that were administered a vaccine of a amphiphilic (AMP) p53 R248Q 30-mer peptide at a concentration of 5 nmol and an adjuvant at a concentration of 10 nmol.

FIG. 23A and FIG. 23B are graphs showing the splenocyte post-dose 2 (PD2) (FIG. 23A), and post-dose 3 (PD3) (FIG. 23B) IFNy ELISpot responses of C57BI6 mice that were administered a vaccine of a soluble (SOL) or amphiphile (AMP) p53 R175H 30-mer peptide at a concentration of 1 .25 nmol and an adjuvant at a concentration of 5 nmol.

FIG. 24A-FIG. 24C are graphs the splenocyte IFNy ELISpot analysis for Granzyme B (FIG. 24A), cytokine FluoroSpot analysis (FIG. 24B), and the percentage of cytokines, including (from top to bottom in each column) IFNy and TNFa, only IFNy, and only TNFa in CD8+ cells (FIG. 21 C) post-dose 3 (PD3) of C57BI6 mice that were administered a vaccine of amphiphile (AMP) p53 R175H 30-mer peptide at a concentration of 5 nmol and an adjuvant at a concentration of 10 nmol.

FIG. 25A- FIG. 25E are graphs showing the concentration of Granzyme B (FIG. 25A), IFNy (FIG. 25B), TNFa (FIG. 25C), GM-CSF (FIG. 25D), and IL2 (FIG. 25E) from splenocytes 7 days after dose 3 for C57BI6 mice that were administered a vaccine of a soluble (SOL) or amphiphilic (AMP) p53 R175H 30- mer peptide at a concentration of 5 nmol and an adjuvant at a concentration of 10 nmol.

FIG. 26A and FIG. 26B are graphs showing the splenocyte post-dose 2 (PD2) (FIG. 26A), and post-dose 3 (PD3) (FIG. 26B) IFNy ELISpot responses of C57BI6 mice that were administered a vaccine of a soluble (SOL) or amphiphile (AMP) p53 R273H 30-mer peptide at a concentration of 1 .25 nmol and an adjuvant at a concentration of 5 nmol.

FIG. 27A-FIG. 27E are graphs showing the concentration of Granzyme B (FIG. 27A), IFNy (FIG. 27B), TNFa (FIG. 27C), GM-CSF (FIG. 27D), and IL2 (FIG. 27E) from splenocytes 7 days after dose 3 for C57BI6 mice that were administered a vaccine of a soluble (SOL) or amphiphilic (AMP) p53 R273H 30- mer peptide at a concentration of 1 .25 nmol and an adjuvant at a concentration of 5 nmol.

FIG. 28 is a graph showing the percentage of target cell killing that occurred after administration of p53 R273H 30-mer peptide at a concentration of 1 .25 nmol and an adjuvant at a concentration of 5 nmol and were analyzed post-dose 4 (PD4).

FIG. 29A-FIG. 29C are graphs showing the splenocyte ELISpot analysis for IFNy (FIG. 29A), the splenocyte ELISpot analysis for Granzyme B (FIG. 29B) and cytokine FluoroSpot analysis for IFNy, TNFa, and IL2 (FIG. 29C) post-dose 3 (PD3) of C57BI6 mice that were administered a vaccine of a soluble (SOL) or amphiphile (AMP) p53 R273C 30-mer peptide at a concentration of 1 .25 nmol and an adjuvant at a concentration of 5 nmol or 5 nmol peptide and an adjuvant at a concentration of 10 nmol, as indicated.

FIG. 30A-FIG. 30E are graphs showing the concentration of Granzyme B (FIG. 30A), IFNy (FIG. 30B), TNFa (FIG. 30C), GM-CSF (FIG. 30D), and IL2 (FIG. 30E) from splenocytes 7 days after dose 3 for C57BI6 mice that were administered a vaccine of a soluble (SOL) or amphiphilic (AMP) p53 R273C 30- mer peptide at a concentration of 5 nmol and an adjuvant at a concentration of 10 nmol.

FIG. 31A-FIG. 31C are graphs showing the splenocyte ELISpot analysis for IFNy (FIG. 31 A), the splenocyte ELISpot analysis for Granzyme B (FIG. 31 B) and the percentage of cytokines, including (from top to bottom in each column) IFNy and TNFa, only IFNy, and only TNFa in CD8+ cells (FIG. 31 C) postdose 3 (PD3) of C57BI6 mice that were administered a vaccine of amphiphile (AMP) p53 R282W 30-mer peptide at a concentration of 5 nmol and an adjuvant at a concentration of 10 nmol.

FIG. 32A-FIG. 32E are graphs showing the concentration of Granzyme B (FIG. 32A), IFNy (FIG. 32B), TNFa (FIG. 32C), GM-CSF (FIG. 32D), and IL2 (FIG. 32E) from splenocytes 7 days after dose 3 for C57BI6 mice that were administered a vaccine of a soluble (SOL) or amphiphilic (AMP) p53 R282W 30- mer peptide at a concentration of 5 nmol and an adjuvant at a concentration of 10 nmol.

FIG. 33A-FIG. 33C are graphs showing the splenocyte post-dose 2 (PD2) (FIG. 33A) and postdose 3 (PD3) (FIG.33B) IFNy ELISpot responses of C57BI6 mice that were administered a vaccine of a SOL or AMP p53 G245S 30-mer peptide at a concentration of 1 .25 nmol and an adjuvant at a concentration of 5 nmol, and PD3 mice that were administered a vaccine of an AMP p53 G245S 30-mer peptide at a concentration of 5 nmol and an adjuvant at a concentration of 5 nmol (FIG. 33C).

FIG. 34A and FIG. 34B are graphs showing the splenocyte ELISpot analysis for Granzyme B (FIG. 34A) and cytokine FluoroSpot analysis (FIG. 34B) post-dose 3 (PD3) of C57BI6 mice that were administered a vaccine of a soluble (SOL) or amphiphile (AMP) p53 G245S 30-mer peptide at a concentration of 5 nmol and an adjuvant at a concentration of 5 nmol.

FIG. 35A-FIG. 35E are graphs showing the concentration of Granzyme B (FIG. 35A), IFNy (FIG. 35B), TNFa (FIG. 35C), GM-CSF (FIG. 35D), and IL2 (FIG. 35E) from splenocytes 7 days after dose 3 for C57BI6 mice that were administered a vaccine of a soluble (SOL) or amphiphilic (AMP) p53 G245S 30- mer peptide at a concentration of 5 nmol and an adjuvant at a concentration of 5 nmol.

FIG. 36A-FIG. 36C are graphs showing the splenocyte ELISpot analysis for IFNy (FIG. 36A), the splenocyte ELISpot analysis for Granzyme B (FIG. 36B) and cytokine FluoroSpot analysis (FIG. 36C) post-dose 3 (PD3) of C57BI6 mice that were administered a vaccine of a soluble (SOL) or amphiphile (AMP) p53 R249S 30-mer peptide at a concentration of 5 nmol and an adjuvant at a concentration of 10 nmol.

FIG. 37A-FIG. 37C are graphs showing the splenocyte ELISpot analysis for IFNy (FIG. 31 A), the splenocyte ELISpot analysis for Granzyme B (FIG. 31 B) and the percentage of cytokines, including (from top to bottom in each column) IFNy and TNFa, only IFNy, and only TNFa in CD8+ cells (FIG. 31 C) postdose 3 (PD3) of C57BI6 mice that were administered a vaccine of amphiphile (AMP) p53 Y220C 30-mer peptide at a concentration of 1 .25 nmol and an adjuvant at a concentration of 5 nmol.

FIG. 38A-FIG. 38E are graphs showing the concentration of Granzyme B (FIG. 38A), IFNy (FIG. 38B), TNFa (FIG. 38C), GM-CSF (FIG. 38D), and IL2 (FIG. 38E) from splenocytes 7 days after dose 3 for C57BI6 mice that were administered a vaccine of a soluble (SOL) or amphiphilic (AMP) p53 Y220C 30- mer peptide at a concentration of 1 .25 nmol and an adjuvant at a concentration of 5 nmol.

FIG. 39A-FIG. 39C are graphs showing the splenocyte ELISpot analysis for IFNy (FIG. 39A), the splenocyte ELISpot analysis for Granzyme B (FIG. 39B) and the percentage of cytokines, including (from top to bottom in each column) IFNy and TNFa, only IFNy, and only TNFa in CD4+ cells (FIG. 39C) postdose 3 (PD3) of C57BI6 mice that were administered a vaccine of amphiphile (AMP) p53 C135Y 30-mer peptide at a concentration of 1 .25 nmol and an adjuvant at a concentration of 5 nmol.

FIG. 40A-FIG. 40E are graphs showing the concentration of Granzyme B (FIG. 40A), IFNy (FIG. 40B), TNFa (FIG. 40C), GM-CSF (FIG. 40D), and IL2 (FIG. 40E) from splenocytes 7 days after dose 3 for C57BI6 mice that were administered a vaccine of a soluble (SOL) or amphiphilic (AMP) p53 C135Y 30- mer peptide at a concentration of 1 .25 nmol and an adjuvant at a concentration of 5 nmol.

FIG. 41 is a graph showing the percentage of target cell killing that occurred after administration of p53 C135Y 30-mer peptide at a concentration of 1 .25 nmol and an adjuvant at a concentration of 5 nmol and were analyzed post-dose 4 (PD4).

FIG. 42A and FIG. 42B are graphs showing the splenocyte post-dose 2 (PD2) (FIG. 42A) and post-dose 3 (PD3) (FIG.42B) IFNy ELISpot responses of C57BI6 mice that were administered a vaccine of a SOL or AMP p53 R158H 30-mer peptide at a concentration of 1 .25 nmol and an adjuvant at a concentration of 5 nmol. FIG. 43A and FIG. 43B are graphs showing the splenocyte post-dose 2 (PD2) (FIG. 43A) and post-dose 3 (PD3) (FIG.43B) IFNy ELISpot responses of C57BI6 mice that were administered a vaccine of a SOL or AMP p53 H214R 30-mer peptide at a concentration of 1 .25 nmol and an adjuvant at a concentration of 5 nmol.

FIG. 44A and FIG. 44B are graphs showing the splenocyte ELISpot analysis for Granzyme B (FIG. 44A) and cytokine FluoroSpot analysis (FIG. 44B) post-dose 3 (PD3) of C57BI6 mice that were administered a vaccine of a soluble (SOL) or amphiphile (AMP) p53 H214R 30-mer peptide at a concentration of 1 .25 nmol and an adjuvant at a concentration of 5 nmol.

FIG. 45A-FIG. 45E are graphs showing the concentration of Granzyme B (FIG. 45A), IFNy (FIG. 45B), TNFa (FIG. 45C), GM-CSF (FIG. 45D), and IL2 (FIG. 45E) from splenocytes 7 days after dose 3 for C57BI6 mice that were administered a vaccine of a soluble (SOL) or amphiphilic (AMP) p53 H214R 30- mer peptide at a concentration of 1 .25 nmol and an adjuvant at a concentration of 5 nmol.

FIG. 46A and FIG. 46B are graphs showing the splenocyte post-dose 2 (PD2) (FIG. 46A) and post-dose 3 (PD3) (FIG.46B) IFNy ELISpot responses of C57BI6 mice that were administered a vaccine of a SOL or AMP p53 wildtype 30-mer peptide at a concentration of 1 .25 nmol and an adjuvant at a concentration of 5 nmol.

FIG. 47A and FIG. 47B are graphs showing the percentage of cytokines, including (from top to bottom in each column) IFNy and TNFa, only IFNy, and only TNFa in CD4+ cells (FIG. 47A) and CD8+ cells (FIG. 47B) found in peripheral blood cells from C57BL/6J mice that were administered a vaccine of a soluble (SOL) or amphiphile (AMP) p53 wildtype 30-mer peptide at a concentration of 1 .25 nmol and an adjuvant at a concentration of 5 nmol.

FIG. 48A-FIG. 48E are graphs showing the concentration of Granzyme B (FIG. 48A), IFNy (FIG. 48B), TNFa (FIG. 48C), GM-CSF (FIG. 48D), and IL2 (FIG. 48E) from splenocytes 7 days after dose 3 for C57BI6 mice that were administered a vaccine of a soluble (SOL) or amphiphilic (AMP) p53 wildtype 30- mer peptide at a concentration of 1 .25 nmol and an adjuvant at a concentration of 5 nmol.

DETAILED DESCRIPTION

Definitions

Terms used in the claims and specification are defined as set forth below unless otherwise specified.

It must be noted that, as used in the specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise.

As used herein, "about" will be understood by persons of ordinary skill and will vary to some extent depending on the context in which it is used. If there are uses of the term which are not clear to persons of ordinary skill given the context in which it is used, "about" will mean up to plus or minus 10% of the particular value.

As used herein, the term "adjuvant" refers to a compound that, with a specific immunogen or antigen, will augment or otherwise alter or modify the resultant immune response. Modification of the immune response includes intensification or broadening the specificity of either or both antibody and cellular immune responses. Modification of the immune response can also mean decreasing or suppressing certain antigen-specific immune responses. In certain embodiments, the adjuvant is a cyclic dinucleotide.

"Amino acid" refers to naturally occurring and synthetic amino acids, as well as amino acid analogs and amino acid mimetics that function in a manner similar to the naturally occurring amino acids. Naturally occurring amino acids are those encoded by the genetic code, as well as those amino acids that are later modified, e.g., hydroxyproline, y-carboxyglutamate, and phosphoserine. Amino acid analogs refers to compounds that have the same basic chemical structure as a naturally occurring amino acid, i.e. , a carbon that is bound to a hydrogen, a carboxyl group, an amino group, and an R group, e.g., homoserine, norleucine, methionine sulfoxide, methionine methyl sulfonium. Such analogs have modified R groups (e.g., norleucine) or modified polypeptide backbones, but retain the same basic chemical structure as a naturally occurring amino acid. Amino acid mimetics refers to chemical compounds that have a structure that is different from the general chemical structure of an amino acid, but that function in a manner similar to a naturally occurring amino acid. Amino acids can be referred to herein by either their commonly known three letter symbols or by the one-letter symbols recommended by the IUPAC-IUB Biochemical Nomenclature Commission. Nucleotides, likewise, can be referred to by their commonly accepted single-letter codes.

An "amino acid substitution" refers to the replacement of at least one existing amino acid residue in a predetermined amino acid sequence (an amino acid sequence of a starting polypeptide) with a second, different "replacement" amino acid residue. An "amino acid insertion" refers to the incorporation of at least one additional amino acid into a predetermined amino acid sequence. While the insertion will usually consist of the insertion of one or two amino acid residues, the present larger "peptide insertions," can be made, e.g., by insertion of about three to about five or even up to about ten, fifteen, or twenty amino acid residues. The inserted residue(s) may be naturally occurring or non-naturally occurring as disclosed above. An "amino acid deletion" refers to the removal of at least one amino acid residue from a predetermined amino acid sequence.

As used herein, "amphiphile" or "amphiphilic" refers to a conjugate comprising a hydrophilic head group and a hydrophobic tail, thereby forming an amphiphilic conjugate. In some embodiments, an amphiphile conjugate comprises a peptide, and one or more hydrophobic lipid tails.

A polypeptide or amino acid sequence "derived from" a designated polypeptide or protein or a "polypeptide fragment" refers to the origin of the polypeptide. Preferably, the polypeptide or amino acid sequence which is derived or is a fragment of is from a particular sequence that has an amino acid sequence that is essentially identical to that sequence or a portion thereof, wherein the portion consists of at least 10-20 amino acids, preferably at least 20-30 amino acids, more preferably at least 30-50 amino acids, or which is otherwise identifiable to one of ordinary skill in the art as having its origin in the sequence. Polypeptides derived from or that are fragments of another polypeptide may have one or more mutations relative to the starting polypeptide, e.g., one or more amino acid residues which have been substituted with another amino acid residue or which has one or more amino acid residue insertions or deletions.

A polypeptide can comprise an amino acid sequence which is not naturally occurring. Such variants necessarily have less than 100% sequence identity or similarity with the starting molecule. In a preferred embodiment, the variant will have an amino acid sequence from about 75% to less than 100% amino acid sequence identity or similarity with the amino acid sequence of the starting polypeptide, more preferably from about 80% to less than 100%, more preferably from about 85% to less than 100%, more preferably from about 90% to less than 100% (e.g., 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%) and most preferably from about 95% to less than 100%, e.g., over the length of the variant molecule.

In one embodiment, there is one amino acid difference between a starting polypeptide sequence and the sequence derived therefrom. Identity or similarity with respect to this sequence is defined herein as the percentage of amino acid residues in the candidate sequence that are identical (i.e., same residue) with the starting amino acid residues, after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent sequence identity.

As used herein, the term "cytotoxic T lymphocyte (CTL) response" refers to an immune response induced by cytotoxic T cells. CTL responses are mediated primarily by CD8+ T cells.

As used herein, the term "effective dose" or "effective dosage" is defined as an amount sufficient to achieve or at least partially achieve the desired effect.

The term "therapeutically effective dose" is defined as an amount sufficient to cure or at least partially arrest the disease and its complications in a patient already suffering from the disease. Amounts effective for this use will depend upon the severity of the disorder being treated and the general state of the patient's own immune system.

As used herein, "immune cell" is a cell of hematopoietic origin and that plays a role in the immune response. Immune cells include lymphocytes (e.g., B cells and T cells), natural killer cells, and myeloid cells (e.g., monocytes, macrophages, eosinophils, mast cells, basophils, and granulocytes). In particular embodiments, the immune cell is T cell.

As used herein, "immune response" refers to a response made by the immune system of an organism to a substance, which includes but is not limited to foreign or self proteins. Three general types of "immune response" include mucosal, humoral, and cellular immune responses. For example, the immune response can include the activation, expansion, and/or increased proliferation of an immune cell. An immune response may also include at least one of the following: cytokine production, T cell activation and/or proliferation, granzyme or perforin production, activation of antigen presenting cells or dendritic cells, antibody production, inflammation, developing immunity, developing hypersensitivity to an antigen, the response of antigen-specific lymphocytes to antigen, clearance of an infectious agent, and transplant or graft rejection.

The terms "inducing an immune response" and "enhancing an immune response" are used interchangeably and refer to the stimulation of an immune response (i.e., either passive or adaptive) to a particular antigen (e.g., a peptide (e.g., the mutant p53 peptide)).

The term "induce" as used with respect to inducing complement dependent cytotoxicity (CDC) or antibody-dependent cellular cytotoxicity (ADCC) refer to the stimulation of particular direct cell killing mechanisms.

As used herein, a subject "in need of prevention," "in need of treatment," or "in need thereof," refers to one, who by the judgment of an appropriate medical practitioner (e.g., a doctor, a nurse, or a nurse practitioner in the case of humans; a veterinarian in the case of non-human mammals), would reasonably benefit from a given treatment (such as treatment with a composition comprising an amphiphilic ligand conjugate). The term "in vivo" refers to processes that occur in a living organism.

The term "in vitro" refers to processes that occur outside a living organism, such as in a test tube, flask, or culture plate.

As used herein, the terms "linked," "operably linked," "fused," or "fusion," are used interchangeably. These terms refer to the joining together of two more elements or components or domains, by an appropriate means including chemical conjugation or recombinant DNA technology. Methods of chemical conjugation (e.g., using heterobifunctional crosslinking agents) are known in the art as are methods of recombinant DNA technology.

The term "lipid" refers to a biomolecule that is soluble in nonpolar solvents and insoluble in water. Lipids are often described as hydrophobic or amphiphilic molecules which allows them to form structures such as vesicles or membranes in aqueous environments. Lipids include fatty acids, glycerolipids, glycerophospholipids, sphingolipids, sterol lipids (including cholesterol), prenol lipids, saccharolipids, and polyketides. In some embodiments, the lipid suitable for the amphiphilic ligand conjugates of the disclosure binds to human serum albumin under physiological conditions. In some embodiments, the lipid suitable for the amphiphilic ligand conjugates of the disclosure inserts into a cell membrane under physiological conditions. In some embodiments, the lipid binds albumin and inserts into a cell membrane under physiological conditions. In some embodiments, the lipid is a diacyl lipid. In some embodiments, the diacyl lipid includes at least 12 carbons. In some embodiments, the diacyl lipid includes 12-30 hydrocarbon units, 14-25 hydrocarbon units, or 16-20 hydrocarbon units. In some embodiments, the diacyl lipid includes 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29 or 30 carbons.

"Nucleic acid" refers to deoxyribonucleotides or ribonucleotides and polymers thereof in either single- or double-stranded form. Unless specifically limited, the term encompasses nucleic acids containing known analogues of natural nucleotides that have similar binding properties as the reference nucleic acid and are metabolized in a manner similar to naturally occurring nucleotides. Unless otherwise indicated, a particular nucleic acid sequence also implicitly encompasses conservatively modified variants thereof (e.g., degenerate codon substitutions) and complementary sequences and as well as the sequence explicitly indicated. Specifically, degenerate codon substitutions can be achieved by generating sequences in which the third position of one or more selected (or all) codons is substituted with mixed-base and/or deoxyinosine residues (Batzer et al., Nucleic Acid Res. 19:5081 , 1991 ; Ohtsuka et al., J. Biol. Chem. 260:2605-2608, 1985); and Cassol et al., 1992; Rossolini et al., Mai. Cell. Probes 8:91 -98, 1994). For arginine and leucine, modifications at the second base can also be conservative. The term nucleic acid is used interchangeably with gene, cDNA, and mRNA encoded by a gene.

Polynucleotides of the present invention can be composed of any polyribonucleotide or polydeoxribonucleotide, which can be unmodified RNA or DNA or modified RNA or DNA. For example, polynucleotides can be composed of single- and double-stranded DNA, DNA that is a mixture of single- and double-stranded regions, single- and double-stranded RNA, and RNA that is mixture of single- and double-stranded regions, hybrid molecules comprising DNA and RNA that can be single-stranded or, more typically, double-stranded or a mixture of single- and double stranded regions. In addition, the polynucleotide can be composed of triple-stranded regions comprising RNA or DNA or both RNA and DNA. A polynucleotide can also contain one or more modified bases or DNA or RNA backbones modified for stability or for other reasons. "Modified" bases include, for example, tritylated bases and unusual bases such as inosine. A variety of modifications can be made to DNA and RNA; thus, "polynucleotide" embraces chemically, enzymatically, or metabolically modified forms. In some embodiments, the polypeptides of the invention are encoded by a nucleotide sequence. Nucleotide sequences of the invention can be useful for a number of applications, including: cloning, gene therapy, protein expression and purification, mutation introduction, DNA vaccination of a host in need thereof, antibody generation for, e.g., passive immunization, PCR, primer and probe generation, and the like.

As used herein, "parenteral administration," "administered parenterally," and other grammatically equivalent phrases, refer to modes of administration other than enteral and topical administration, usually by injection, and include, without limitation, intravenous, intranasal, intraocular, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal, epidural, intracerebral, intracranial, intracarotid and intrasternal injection and infusion.

As generally used herein, "pharmaceutically acceptable" refers to those compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues, organs, and/or bodily fluids of human beings and animals without excessive toxicity, irritation, allergic response, or other problems or complications commensurate with a reasonable benef it/risk ratio.

The term “pharmaceutically acceptable salt,” as used herein, means any pharmaceutically acceptable salt of a conjugate, oligonucleotide, or peptide disclosed herein. Pharmaceutically acceptable salts of any of the compounds and nucleic acid sequences described herein may include those that are within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and animals without undue toxicity, irritation, allergic response and are commensurate with a reasonable benef it/risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, pharmaceutically acceptable salts are described in: Berge et al., J. Pharmaceutical Sciences 66:1 -19, 1977 and in Pharmaceutical Salts: Properties, Selection, and Use (Eds. P.H. Stahl and C.G. Wermuth), Wiley-VCH, 2008. The salts can be prepared in situ during the final isolation and purification of the compounds described herein or separately by reacting a free base group with a suitable acid. Representative acid addition salts include acetate, adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, fumarate, glucoheptonate, glycerophosphate, hemisulfate, heptonate, hexanoate, hydrobromide, hydrochloride, hydroiodide, 2- hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, toluenesulfonate, undecanoate, valerate salts, and the like. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like, as well as nontoxic ammonium, quaternary ammonium, and amine cations, including, but not limited to ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, ethylamine, and the like. References to the compounds, nucleic acids, conjugates, oligonucleotides, or polypeptides in the claims and elsewhere herein optionally include pharmaceutically acceptable salts thereof unless otherwise indicated or not applicable. As used herein, the term “physiological conditions” refers to the in vivo condition of a subject. In some embodiments, physiological condition refers to a neutral pH (e.g., pH between 6-8).

As used herein, the term “peptide” refers to a polymer having 30 or fewer amino acid residues. "Polypeptide," "peptide", and "protein" are used interchangeably herein to refer to a polymer of amino acid residues. The terms apply to amino acid polymers in which one or more amino acid residue is an artificial chemical mimetic of a corresponding naturally occurring amino acid, as well as to naturally occurring amino acid polymers and non-naturally occurring amino acid polymer.

As used herein, the term "subject" or "mammal" or "patient" includes any human or non-human animal. For example, the methods and compositions of the present invention can be used to treat a subject with a disease or condition. The term "non-human animal" includes all vertebrates, e.g., mammals and non-mammals, such as non-human primates, sheep, dogs, cats, mice, horses, pigs, cows, chickens, amphibians, reptiles, etc.

The term "sufficient amount" or "amount sufficient to" means an amount sufficient to produce a desired effect, e.g., an amount sufficient to reduce the diameter of a tumor.

The term "T cell" refers to a type of white blood cell that can be distinguished from other white blood cells by the presence of a T cell receptor on the cell surface. There are several subsets of T cells, including, but not limited to, T helper cells ( a.k.a. TH cells or CD4⁺ T cells) and subtypes, including TH, TH2, TH3, TH17, TH9, and TFH cells, cytotoxic T cells (i.e., Tc cells, CD8⁺ T cells, cytotoxic T lymphocytes, T-killer cells, killer T cells), memory T cells and subtypes, including central memory T cells (TCM cells), effector memory T cells (TEM and TEMRA cells), and resident memory T cells (TRM cells), regulatory T cells (a.k.a. Treg cells or suppressor T cells) and subtypes, including CD4⁺ FOXP3⁺ T_reg cells, CD4+FOXP3- Treg cells, Tr1 cells, Th3 cells, and T_reg17 cells, natural killer T cells (a.k.a. NKT cells), mucosal associated invariant T cells (MAITs), and gamma delta T cells (yb T cells), including Vy9/V52 T cells. Any one or more of the aforementioned or unmentioned T cells may be the target cell type for a method of use of the invention.

The terms "treat," "treating," and "treatment," as used herein, refer to therapeutic or preventative measures described herein. The methods of "treatment" employ administration to a subject, in need of such treatment, a peptide and an albumin-binding domain of the present disclosure. In some embodiments, the mutant p53 peptide conjugated to an albumin-binding domain is administered to a subject in need of an enhanced immune response against a particular antigen or a subject who ultimately may acquire such a disorder, in order to prevent, cure, delay, reduce the severity of, or ameliorate one or more symptoms of the disorder or recurring disorder, or in order to prolong the survival of a subject beyond that expected in the absence of such treatment.

As used herein, "vaccine" refers to a formulation which contains an amphiphilic construct described herein, optionally combined with an adjuvant, which is in a form that is capable of being administered to a vertebrate and which induces a protective immune response sufficient to induce immunity to prevent and/or ameliorate a disease or condition and/or to reduce at least one symptom of a disease or condition. Typically, the vaccine comprises a conventional saline or buffered aqueous solution medium in which a composition as described herein is suspended or dissolved. In this form, a composition as described herein is used to prevent, ameliorate, or otherwise treat an infection or disease. Upon introduction into a host, the vaccine provokes an immune response including, but not limited to, the inducing a protective immune response to induce immunity to prevent and/or ameliorate a disease or condition and/or to reduce at least one symptom of a disease or condition.

Peptides

Described herein are compounds including a peptide, wherein the peptide is a p53 peptide or a mutant p53 peptide. The peptide is conjugated to an albumin-binding domain optionally by way of a linker.

In some embodiments, the peptide is a wild-type p53 peptide. In some embodiments, the wildtype p53 peptide includes an amino acid residue that is a mutation hot spot. The wild type p53 sequence with amino acid residues in bold that are mutation hot spots is shown below:

1 MEEPQSDPSV EPPLSQETFS DLWKLLPENN VLSPLPSQAM DDLMLSPDDI EQWFTEDPGP 61 DEAPRMPEAA PPVAPAPAAP TPAAPAPAPS WPLSSSVPSQ KTYQGSYGFR LGFLHSGTAK 121 SVTCTYSPAL NKMFCQLAKT CPVQLWVDST PPPGTRVRAM AIYKQSQHMT EVVRRCPHHE 181 RCSDSDGLAP PQHLIRVEGN LRVEYLDDRN TFRHSVVVPY EPPEVGSDCT TIHYNYMCNS 241 SCMGGMNRRP ILTIITLEDS SGNLLGRNSF EVRVCACPGR DRRTEEENLR KKGEPHHELP 301 PGSTKRALPN NTSSSPQPKK KPLDGEYFTL QIRGRERFEM FRELNEALEL KDAQAGKEPG 361 GSRAHSSHLK SKKGQSTSRH KKLMFKTEGP DSD (SEQ ID NO: 1 )

In some embodiments, the wild-type p53 peptide comprises a 5 to 50 (e.g., 5, 6, 7, 8, 9, 11 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 35, 36, 37, 38, 39, 40,

41 , 42, 43, 44, 45, 46, 47, 48, 49, or 50) amino acid fragment of SEQ ID NO: 1 . In some embodiments, the wild-type p53 peptide comprises a 5 to 50 (e.g., 5, 6, 7, 8, 9, 11 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 ,

22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 35, 36, 37, 38, 39, 40, 41 , 42, 43, 44, 45, 46, 47, 48, 49, or 50) amino acid fragment of SEQ ID NO: 1 and is 6 to 60 (e.g. 6, 7, 8 9, 10, 11 , 12, 13, 15 15, 16 ,17,

18, 19, 20 , 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 35, 36, 37, 38, 39, 40, 41 , 42, 43, 44, 45, 46, 47, 48, 49, 50, 51 , 52, 53, 54, 55, 56, 57, 58, 59, or 60) amino acids in length. In some embodiments, the wild-type p53 peptide is a 5 to 50 (e.g., 5, 6, 7, 8, 9, 11 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 35, 36, 37, 38, 39, 40, 41 , 42, 43, 44, 45, 46, 47, 48, 49, or 50) amino acid fragment of SEQ ID NO: 1 . In some embodiments, the wild-type p53 peptide is a 20 to 40 (e.g., 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 35, 36, 37, 38, 39, or 40) amino acid fragment of SEQ ID NO: 1 . In some embodiments, the wild-type p53 peptide is a 20 to 30 (e.g., 21 , 22,

23, 24, 25, 26, 27, 28, 29, or 30) amino acid fragment of SEQ ID NO: 1 . In some embodiments, the wildtype p53 peptide is a 30 amino acid fragment of SEQ ID NO: 1 .

In some embodiments, the wild-type p53 peptide has the sequence MAIYKQSQHMTEVVRRCPHHERCSDSDGLAP (SEQ ID NQ:100). The mutation hot spot residue is bolded. In some embodiments, the wild-type p53 peptide has the sequence AIYKQSQHM (SEQ ID NO:101 ), which has been shown to be immunogenic and to elicit T cell responses.

In some embodiments, the wild-type p53 peptide has the sequence EGNLRVEYLDDRNTFRHSVVVPCEPPEVGSD (SEQ ID NO:102). The mutation hot spot residue is bolded. In some embodiments, the wild-type p53 peptide has the sequence EYLDDRNTF (SEQ ID NO:103), which has been shown to be immunogenic and to elicit T cell responses. In some embodiments, the wild-type p53 peptide has the sequence YLDDRNTFRHSVVVPYEPPEVGSDCTTIHYN (SEQ ID NQ:104). The mutation hot spot residue is bolded. In some embodiments, the wild-type p53 peptide has the sequence VVPYEPPEV (SEQ ID NO:105), which has been shown to be immunogenic and to elicit T cell responses.

In some embodiments, the wild-type p53 peptide has the sequence TTIHYNYMCNSSCMGGMNRRPILTIITLEDSSGNL (SEQ ID NQ:106). The mutation hot spot residues are bolded.

In some embodiments, the wild-type p53 peptide has the sequence EDSSGNLLGRNSFEVRVCACPGRDRRTEEENLRKK (SEQ ID NQ:107). The mutation hot spot residues are bolded. In some embodiments, the wild-type p53 peptide has the sequence LLGRNSFEV (SEQ ID NQ:108), which has been shown to be immunogenic and to elicit T cell responses.

In some embodiments, the wild-type p53 peptide has the sequence RLGFLHSGTAKSVTC (SEQ ID NQ:109).

In some embodiments, the wild-type p53 peptide has the sequence STPPPGTRV (SEQ ID NQ:110).

In some embodiments, the wild-type p53 peptide has the sequence TYPALNKMF (SEQ ID NO:111 ).

In some embodiments, the wild-type p53 peptide has the sequence RMPEAAPPV (SEQ ID NO:112).

In some embodiments, the wild-type p53 peptide has the sequence TEDPGPDEAPRMPEAAPPVAPAPAAPTPAA (SEQ ID NO: 121 ).

In some embodiments, the peptide is a mutant p53 peptide. The mutant p53 peptide comprises a fragment of the wild-type p53 polypeptide having the amino acid sequence of SEQ ID NO: 1 , wherein the mutant p53 peptide includes one or more amino acid substitutions in comparison to the wildtype p53 polypeptide. In some embodiments, the wild-type p53 peptide comprises a 5 to 50 (e.g., 5, 6, 7, 8, 9, 11 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 35, 36, 37, 38, 39, 40, 41 , 42, 43, 44, 45, 46, 47, 48, 49, or 50) amino acid fragment of SEQ ID NO: 1 including at least one amino acid substitution relative to the corresponding sequence of SEQ ID NO:1 . In some embodiments, the wild-type p53 peptide comprises a 5 to 50 (e.g., 5, 6, 7, 8, 9, 11 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 35, 36, 37, 38, 39, 40, 41 , 42, 43, 44, 45, 46, 47, 48, 49, or 50) amino acid fragment of SEQ ID NO: 1 including at least one amino acid substitution relative to the corresponding sequence of SEQ ID NO:1 and is 6 to 60 (e.g. 6, 7, 8 9, 10, 11 , 12, 13, 15 15, 16 ,17, 18, 19, 20 , 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 35, 36, 37, 38, 39, 40, 41 , 42, 43, 44, 45, 46, 47, 48, 49, 50, 51 , 52, 53, 54, 55, 56, 57, 58, 59, or 60) amino acids in length. In some embodiments, the mutant p53 peptide is a fragment having 5 and 20 amino acids (e.g., 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, or 25 amino acids) of SEQ ID NO: 1 including at least one amino acid substitution relative to the corresponding sequence of SEQ ID NO:1 . In some embodiments, the mutant p53 peptide is a 10 amino acid fragment of SEQ ID NO: 1 including at least one amino acid substitution relative to the corresponding sequence of SEQ ID NO:1 . In some embodiments, the mutant p53 peptide is a 15 or 18 amino acid fragment of SEQ ID NO: 1 including at least one amino acid substitution relative to the corresponding sequence of SEQ ID NO:1 . In some embodiments, the p53 mutant peptide comprises a fragment of SEQ ID NO: 1 having an amino acid substitution at the 248^th amino acid from the N-terminus, which is an Arg residue. For example, the p53 mutant peptide may include an amino acid substitution at position 248 of SEQ ID NO: 1 wherein the Arg residue is substituted for a Gin residue (R248Q) or wherein the Arg residue is substituted for a Trp residue (R248W). In some embodiments, the mutant p53 peptide consists of the amino acid sequence of YMCNSSCMGGMNWRPILTIITLEDS (SEQ ID NO: 2), or a fragment thereof. In some embodiments, the mutant p53 peptide consists of the amino acid sequence of YMCNSSCMGGMNQRPILTIITLEDS (SEQ ID NO: 3), or a fragment thereof. In some embodiments, the mutant p53 peptide consists of the amino acid sequence of CNWRPILTIIT (SEQ ID NO: 123), or a fragment thereof. In some embodiments, the mutant p53 peptide consists of the amino acid sequence of CNSSCMGGMNWRPILTIIT (SEQ ID NO: 29), or a fragment thereof. In some embodiments, the mutant p53 peptide consists of the amino acid sequence of CYNYMCNSSCMGGMNWRPILTIITLEDSSGN (SEQ ID NO: 124), or a fragment thereof. In some embodiments, the mutant p53 peptide consists of the amino acid sequence of CNQRPILTIIT (SEQ ID NO: 125), or a fragment thereof. In some embodiments, the mutant p53 peptide consists of the amino acid sequence of CYNYMCNSSCMGGMNQRPILTIITLEDSSGN (SEQ ID NO: 126), or a fragment thereof. In some embodiments, the mutant p53 peptide consists of the amino acid sequence of CNSSCMGGMNQRPILTIIT (SEQ ID NO: 38), or a fragment thereof. In some embodiments, the mutant p53 peptide consists of the amino acid sequence of YNYMCNSSCMGGMNWRPILTIITLEDSSGN (SEQ ID NO: 127), or fragment thereof.

In some embodiments, the mutant p53 peptide consists of the amino acid sequence of YNYMCNSSCMGGMNQRPILTIITLEDSSGN (SEQ ID NO: 128), or fragment thereof.

In some embodiments, the p53 mutant peptide comprises a fragment of SEQ ID NO: 1 having an amino acid substitution at position 273 of SEQ ID NO: 1 wherein the Arg residue is substituted for a His residue (R273H) or wherein the Arg residue is substituted for a Cys residue (R273C). In some embodiments, the mutant p53 peptide consists of the amino acid sequence DSSGNLLGRNSFEVHVCACPGRDRRTEEEN (SEQ ID NO: 99), or a fragment thereof. In some embodiments, the mutant p53 peptide consists of the amino acid sequence DSSGNLLGRNSFEVCVCACPGRDRRTEEEN (SEQ ID NO: 113), or a fragment thereof.

In some embodiments, the p53 mutant peptide comprises a fragment of SEQ ID NO: 1 having an amino acid substitution at position 282 of SEQ ID NO: 1 wherein the Arg residue is substituted for a Trp residue (R282W). In some embodiments, the mutant p53 peptide consists of the amino acid sequence NSFEVCVCACPGRDWRTEEENLRKKGEPHH (SEQ ID NO: 114), or a fragment thereof. In some embodiments, the mutant p53 peptide consists of the amino acid sequence NSFEVRVCACPGRDWRTEEENLRKKGEPHH (SEQ ID NO: 129), or a fragment thereof.

In some embodiments, the p53 mutant peptide comprises a fragment of SEQ ID NO: 1 having an amino acid substitution at position 245 of SEQ ID NO: 1 wherein the Gly residue is substituted for a Ser residue (G245S). In some embodiments, the mutant p53 peptide consists of the amino acid sequence TIHYNYMCNSSCMGSMNRRPILTIITLEDS (SEQ ID NO: 115), or a fragment thereof.

In some embodiments, the p53 mutant peptide comprises a fragment of SEQ ID NO: 1 having an amino acid substitution at position 249 of SEQ ID NO: 1 wherein the Arg residue is substituted for a Ser residue (R249S). In some embodiments, the mutant p53 peptide consists of the amino acid sequence NYMCNSSCMGGMNRSPILTIITLEDSSGNL (SEQ ID NO: 116), or a fragment thereof.

In some embodiments, the p53 mutant peptide comprises a fragment of SEQ ID NO: 1 having an amino acid substitution at position 220 of SEQ ID NO: 1 wherein the Tyr residue is substituted for a Cys residue (Y220C). In some embodiments, the mutant p53 peptide consists of the amino acid sequence LDDRNTFRHSVVVPCEPPEVGSDCTTIHYN (SEQ ID NO: 117), or a fragment thereof.

In some embodiments, the p53 mutant peptide comprises a fragment of SEQ ID NO: 1 having an amino acid substitution at position 135 of SEQ ID NO: 1 wherein the Cys residue is substituted for a Tyr residue (C135Y). In some embodiments, the mutant p53 peptide consists of the amino acid sequence RLGFLHSGTAKSVTCTYSPALNKMFYQLAK (SEQ ID NO: 118), or a fragment thereof.

In some embodiments, the p53 mutant peptide comprises a fragment of SEQ ID NO: 1 having an amino acid substitution at position 158 of SEQ ID NO: 1 wherein the Arg residue is substituted for a His residue (R158H). In some embodiments, the mutant p53 peptide consists of the amino acid sequence PVQLWVDSTPPPGTRVHAMAIYKQSQHMTE (SEQ ID NO: 119), or a fragment thereof.

In some embodiments, the p53 mutant peptide comprises a fragment of SEQ ID NO: 1 having an amino acid substitution at position 214 of SEQ ID NO: 1 wherein the His residue is substituted for an Arg residue (H214R). In some embodiments, the mutant p53 peptide consists of the amino acid sequence LRVEYLDDRNTFRRSVVVPYEPPEVGSDCT (SEQ ID NO: 120), or a fragment thereof.

In some embodiments, the p53 mutant peptide comprises a fragment of SEQ ID NO: 1 having an amino acid substitution at position 175 of SEQ ID NO: 1 wherein the Arg residue is substituted for a His residue (R175H). In some embodiments, the mutant p53 peptide consists of the amino acid sequence RAMAIYKQSQHMTEVVRHCPHHERCSDSDG (SEQ ID NO: 122), or a fragment thereof.

The mutant p53 peptide may include a 10 amino acid fragment of SEQ ID NO: 2. The mutant p53 peptide may include a 10 amino acid fragment of SEQ ID NO: 3. The mutant p53 peptide may include a 10 amino acid fragment of SEQ ID NO: 99. The mutant p53 peptide may include a 10 amino acid fragment of SEQ ID NO: 113. The mutant p53 peptide may include a 10 amino acid fragment of SEQ ID NO:114. The mutant p53 peptide may include a 10 amino acid fragment of SEQ ID NO:115. The mutant p53 peptide may include a 10 amino acid fragment of SEQ ID NO: 116. The mutant p53 peptide may include a 10 amino acid fragment of SEQ ID NO:117. The mutant p53 peptide may include a 10 amino acid fragment of SEQ ID NO: 118. The mutant p53 peptide may include a 10 amino acid fragment of SEQ ID NO: 119. The mutant p53 peptide may include a 10 amino acid fragment of SEQ ID NO: 120. The mutant p53 peptide may include a 10 amino acid fragment of SEQ ID NO: 122. The mutant p53 peptide may include a 12 amino acid fragment of SEQ ID NO: 2. The mutant p53 peptide may include a 12 amino acid fragment of SEQ ID NO: 3. The mutant p53 peptide may include a 12 amino acid fragment of SEQ ID NO: 99. The mutant p53 peptide may include a 12 amino acid fragment of SEQ ID NO: 113. The mutant p53 peptide may include a 12 amino acid fragment of SEQ ID NO: 114. The mutant p53 peptide may include a 12 amino acid fragment of SEQ ID NO: 115. The mutant p53 peptide may include a 12 amino acid fragment of SEQ ID NO: 116. The mutant p53 peptide may include a 12 amino acid fragment of SEQ ID NO: 117. The mutant p53 peptide may include a 12 amino acid fragment of SEQ ID NO: 118. The mutant p53 peptide may include a 12 amino acid fragment of SEQ ID NO: 119. The mutant p53 peptide may include a 12 amino acid fragment of SEQ ID NO: 120. The mutant p53 peptide may include a 12 amino acid fragment of SEQ ID NO: 122. The mutant p53 peptide may include a 14 amino acid fragment of SEQ ID NO: 2. The mutant p53 peptide may include a 14 amino acid fragment of SEQ ID NO: 3. The mutant p53 peptide may include a 14 amino acid fragment of SEQ ID NO: 99. The mutant p53 peptide may include a 14 amino acid fragment of SEQ ID NO: 113. The mutant p53 peptide may include a 14 amino acid fragment of SEQ ID NO: 114. The mutant p53 peptide may include a 14 amino acid fragment of SEQ ID NO: 115. The mutant p53 peptide may include a 14 amino acid fragment of SEQ ID NO: 116. The mutant p53 peptide may include a 14 amino acid fragment of SEQ ID NO: 117. The mutant p53 peptide may include a 14 amino acid fragment of SEQ ID NO: 118. The mutant p53 peptide may include a 14 amino acid fragment of SEQ ID NO: 119. The mutant p53 peptide may include a 14 amino acid fragment of SEQ ID NO: 120. The mutant p53 peptide may include a 14 amino acid fragment of SEQ ID NO: 122. The mutant p53 peptide may include a 16 amino acid fragment of SEQ ID NO: 2. The mutant p53 peptide may include a 16 amino acid fragment of SEQ ID NO: 3. The mutant p53 peptide may include a 16 amino acid fragment of SEQ ID NO: 99. The mutant p53 peptide may include a

16 amino acid fragment of SEQ ID NO: 113. The mutant p53 peptide may include a 16 amino acid fragment of SEQ ID NO: 114. The mutant p53 peptide may include a 16 amino acid fragment of SEQ ID NO: 115. The mutant p53 peptide may include a 16 amino acid fragment of SEQ ID NO: 116. The mutant p53 peptide may include a 16 amino acid fragment of SEQ ID NO: 117. The mutant p53 peptide may include a 16 amino acid fragment of SEQ ID NO: 118. The mutant p53 peptide may include a 16 amino acid fragment of SEQ ID NO: 119. The mutant p53 peptide may include a 16 amino acid fragment of SEQ ID NO: 120. The mutant p53 peptide may include a 16 amino acid fragment of SEQ ID NO: 122. The mutant p53 peptide may include a 17 amino acid fragment of SEQ ID NO: 2. The mutant p53 peptide may include a 17 amino acid fragment of SEQ ID NO: 3. The mutant p53 peptide may include a

17 amino acid fragment of SEQ ID NO: 99. The mutant p53 peptide may include a 17 amino acid fragment of SEQ ID NO: 113. The mutant p53 peptide may include a 17 amino acid fragment of SEQ ID NO: 114. The mutant p53 peptide may include a 17 amino acid fragment of SEQ ID NO: 115. The mutant p53 peptide may include a 17 amino acid fragment of SEQ ID NO: 116. The mutant p53 peptide may include a 17 amino acid fragment of SEQ ID NO: 117. The mutant p53 peptide may include a 17 amino acid fragment of SEQ ID NO: 118. The mutant p53 peptide may include a 17 amino acid fragment of SEQ ID NO: 119. The mutant p53 peptide may include a 17 amino acid fragment of SEQ ID NO: 120. The mutant p53 peptide may include a 17 amino acid fragment of SEQ ID NO: 122. The mutant p53 peptide may include an 18 amino acid fragment of SEQ ID NO: 2. The mutant p53 peptide may include an 18 amino acid fragment of SEQ ID NO: 3. The mutant p53 peptide may include an 18 amino acid fragment of SEQ ID NO: 99. The mutant p53 peptide may include an 18 amino acid fragment of SEQ ID NO: 113. The mutant p53 peptide may include an 18 amino acid fragment of SEQ ID NO: 114. The mutant p53 peptide may include an 18 amino acid fragment of SEQ ID NO: 115. The mutant p53 peptide may include an 18 amino acid fragment of SEQ ID NO: 116. The mutant p53 peptide may include an 18 amino acid fragment of SEQ ID NO: 117. The mutant p53 peptide may include an 18 amino acid fragment of SEQ ID NO: 118. The mutant p53 peptide may include an 18 amino acid fragment of SEQ ID NO: 119. The mutant p53 peptide may include an 18 amino acid fragment of SEQ ID NO: 120. The mutant p53 peptide may include an 18 amino acid fragment of SEQ ID NO: 122. The mutant p53 peptide may include a 19 amino acid fragment of SEQ ID NO: 2. The mutant p53 peptide may include a 19 amino acid fragment of SEQ ID NO: 3. The mutant p53 peptide may include a 19 amino acid fragment of SEQ ID NO: 99. The mutant p53 peptide may include a 19 amino acid fragment of SEQ ID NO: 113. The mutant p53 peptide may include a 19 amino acid fragment of SEQ ID NO: 114. The mutant p53 peptide may include a 19 amino acid fragment of SEQ ID NO: 115. The mutant p53 peptide may include a 19 amino acid fragment of SEQ ID NO: 116. The mutant p53 peptide may include a 19 amino acid fragment of SEQ ID NO: 117. The mutant p53 peptide may include a 19 amino acid fragment of SEQ ID NO: 118. The mutant p53 peptide may include a 19 amino acid fragment of SEQ ID NO: 119. The mutant p53 peptide may include a 19 amino acid fragment of SEQ ID NO: 120. The mutant p53 peptide may include a 19 amino acid fragment of SEQ ID NO: 122. The mutant p53 peptide may include a 20 amino acid fragment of SEQ ID NO: 2. The mutant p53 peptide may include a 20 amino acid fragment of SEQ ID NO: 3. The mutant p53 peptide may include a 20 amino acid fragment of SEQ ID NO: 99. The mutant p53 peptide may include a 20 amino acid fragment of SEQ ID NO: 113. The mutant p53 peptide may include a 20 amino acid fragment of SEQ ID NO: 114. The mutant p53 peptide may include a 20 amino acid fragment of SEQ ID NO: 115. The mutant p53 peptide may include a 20 amino acid fragment of SEQ ID NO: 116. The mutant p53 peptide may include a 20 amino acid fragment of SEQ ID NO: 117. The mutant p53 peptide may include a 20 amino acid fragment of SEQ ID NO: 118. The mutant p53 peptide may include a 20 amino acid fragment of SEQ ID NO: 119. The mutant p53 peptide may include a 20 amino acid fragment of SEQ ID NO: 120. The mutant p53 peptide may include a 20 amino acid fragment of SEQ ID NO: 122. The mutant p53 peptide may include a 25 amino acid fragment of SEQ ID NO: 2. The mutant p53 peptide may include a 25 amino acid fragment of SEQ ID NO: 3. The mutant p53 peptide may include a 25 amino acid fragment of SEQ ID NO: 99. The mutant p53 peptide may include a 25 amino acid fragment of SEQ ID NO: 113. The mutant p53 peptide may include a 25 amino acid fragment of SEQ ID NO: 114. The mutant p53 peptide may include a 25 amino acid fragment of SEQ ID NO: 115. The mutant p53 peptide may include a 25 amino acid fragment of SEQ ID NO: 116. The mutant p53 peptide may include a 25 amino acid fragment of SEQ ID NO: 117. The mutant p53 peptide may include a 25 amino acid fragment of SEQ ID NO: 118. The mutant p53 peptide may include a 25 amino acid fragment of SEQ ID NO: 119. The mutant p53 peptide may include a 25 amino acid fragment of SEQ ID NO: 120. The mutant p53 peptide may include a 25 amino acid fragment of SEQ ID NO: 122. The mutant p53 peptide may include a 29 amino acid fragment of SEQ ID NO: 2. The mutant p53 peptide may include a 29 amino acid fragment of SEQ ID NO: 3. The mutant p53 peptide may include a 29 amino acid fragment of SEQ ID NO: 99. The mutant p53 peptide may include a 29 amino acid fragment of SEQ ID NO: 113. The mutant p53 peptide may include a 29 amino acid fragment of SEQ ID NO: 114. The mutant p53 peptide may include a 29 amino acid fragment of SEQ ID NO: 115. The mutant p53 peptide may include a 29 amino acid fragment of SEQ ID NO: 116. The mutant p53 peptide may include a 29 amino acid fragment of SEQ ID NO: 117. The mutant p53 peptide may include a 29 amino acid fragment of SEQ ID NO: 118. The mutant p53 peptide may include a 29 amino acid fragment of SEQ ID NO: 119. The mutant p53 peptide may include a 29 amino acid fragment of SEQ ID NO: 120. The mutant p53 peptide may include a 29 amino acid fragment of SEQ ID NO: 122. In some embodiments, the mutant p53 peptide includes a 15 amino acid fragment of SEQ ID NO: 2. In some embodiments, the mutant p53 peptide includes a 15 amino acid fragment of SEQ ID NO: 3. In some embodiments, the mutant p53 peptide includes a 15 amino acid fragment of SEQ ID NO: 99. In some embodiments, the mutant p53 peptide includes a 15 amino acid fragment of SEQ ID NO: 1 13. In some embodiments, the mutant p53 peptide includes a 1 5 amino acid fragment of SEQ ID NO: 1 14. In some embodiments, the mutant p53 peptide includes a 1 5 amino acid fragment of SEQ ID NO: 1 15. In some embodiments, the mutant p53 peptide includes a 1 5 amino acid fragment of SEQ ID NO: 1 16. In some embodiments, the mutant p53 peptide includes a 1 5 amino acid fragment of SEQ ID NO: 1 17. In some embodiments, the mutant p53 peptide includes a 1 5 amino acid fragment of SEQ ID NO: 1 18. In some embodiments, the mutant p53 peptide includes a 1 5 amino acid fragment of SEQ ID NO: 1 19. In some embodiments, the mutant p53 peptide includes a 1 5 amino acid fragment of SEQ ID NO: 120. In some embodiments, the mutant p53 peptide includes a 1 5 amino acid fragment of SEQ ID NO: 122. In some embodiments, the mutant p53 peptide includes an 18 amino acid fragment of SEQ ID NO: 2. In some embodiments, the mutant p53 peptide includes an 18 amino acid fragment of SEQ ID NO: 3. In some embodiments, the mutant p53 peptide includes an 18 amino acid fragment of SEQ ID NO: 99. In some embodiments, the mutant p53 peptide includes an 18 amino acid fragment of SEQ ID NO: 1 13. In some embodiments, the mutant p53 peptide includes an 18 amino acid fragment of SEQ ID NO: 1 14. In some embodiments, the mutant p53 peptide includes an 18 amino acid fragment of SEQ ID NO: 1 15. In some embodiments, the mutant p53 peptide includes an 18 amino acid fragment of SEQ ID NO: 1 16. In some embodiments, the mutant p53 peptide includes an 18 amino acid fragment of SEQ ID NO: 1 17. In some embodiments, the mutant p53 peptide includes an 18 amino acid fragment of SEQ ID NO: 1 18. In some embodiments, the mutant p53 peptide includes an 18 amino acid fragment of SEQ ID NO: 1 19. In some embodiments, the mutant p53 peptide includes an 18 amino acid fragment of SEQ ID NO: 120. In some embodiments, the mutant p53 peptide includes an 18 amino acid fragment of SEQ ID NO: 122. In some embodiments, the 10 amino acid fragment of SEQ ID NO: 2 comprises or consists of the sequence NWRPILTIIT (SEQ ID NO: 46). In some embodiments, the 10 amino acid fragment of SEQ ID NO: 3 comprises or consists of the sequence NQRPILTIIT (SEQ ID NO: 47). In some embodiments, the 18 amino acid fragment of SEQ ID NO: 2 comprises or consists of the sequence

NSSCMGGMNWRPILTIIT (SEQ ID NO: 48). In some embodiments, the 18 amino acid fragment of SEQ ID NO: 3 comprises or consists of the sequence NSSCMGGMNQRPILTIIT (SEQ ID NO: 49).

The mutant p53 peptide comprises a fragment of any one of the mutant p53 polypeptides described in Table 1 . In some embodiments, the p53 mutant peptide comprises a fragment of SEQ ID NO: 1 having an amino acid substitution at the 135^th amino acid from the N-terminus, which is a Cys residue. For example, the p53 mutant peptide may include an amino acid substitution at position 135 of SEQ ID NO: 1 wherein the Cys residue is substituted for a Tyr residue (C135Y). In some embodiments, the p53 mutant peptide comprises a fragment of SEQ ID NO: 1 having an amino acid substitution at the 141^th amino acid from the N-terminus, which is a Cys residue. For example, the p53 mutant peptide may include an amino acid substitution at position 141 of SEQ ID NO: 1 wherein the Cys residue is substituted for a Tyr residue (C141 Y). In some embodiments, the p53 mutant peptide comprises a fragment of SEQ ID NO: 1 having an amino acid substitution at the 151 ^st amino acid from the N-terminus, which is a Pro residue. For example, the p53 mutant peptide may include an amino acid substitution at position 151 of SEQ ID NO: 1 wherein the Pro residue is substituted for a Ser residue (P151 S). In some embodiments, the p53 mutant peptide comprises a fragment of SEQ ID NO: 1 having an amino acid substitution at the 152^nd amino acid from the N-terminus, which is a Pro residue. For example, the p53 mutant peptide may include an amino acid substitution at position 152 of SEQ ID NO: 1 wherein the Pro residue is substituted for a Leu residue (P152L). In some embodiments, the p53 mutant peptide comprises a fragment of SEQ ID NO: 1 having an amino acid substitution at the 157^th amino acid from the N-terminus, which is a Vai residue. For example, the p53 mutant peptide may include an amino acid substitution at position 157 of SEQ ID NO: 1 wherein the Vai residue is substituted for a Phe residue (V157F). In some embodiments, the p53 mutant peptide comprises a fragment of SEQ ID NO: 1 having an amino acid substitution at the 158^th amino acid from the N-terminus, which is an Arg residue. For example, the p53 mutant peptide may include an amino acid substitution at position 158 of SEQ ID NO: 1 wherein the Arg residue is substituted for a His residue (R158H) or the Arg residue is substituted for a Leu residue (R158L). In some embodiments, the p53 mutant peptide comprises a fragment of SEQ ID NO: 1 having an amino acid substitution at the 161 ^st amino acid from the N-terminus, which is an Ala residue. For example, the p53 mutant peptide may include an amino acid substitution at position 161 of SEQ ID NO: 1 wherein the Ala residue is substituted for a Thr residue (A161 T). In some embodiments, the p53 mutant peptide comprises a fragment of SEQ ID NO: 1 having an amino acid substitution at the 163^rd amino acid from the N-terminus, which is a Tyr residue. For example, the p53 mutant peptide may include an amino acid substitution at position 163 of SEQ ID NO: 1 wherein the Tyr residue is substituted for a Cys residue (Y163C). In some embodiments, the p53 mutant peptide comprises a fragment of SEQ ID NO: 1 having an amino acid substitution at the 173^rd amino acid from the N-terminus, which is a Vai residue. For example, the p53 mutant peptide may include an amino acid substitution at position 173 of SEQ ID NO: 1 wherein the Vai residue is substituted for a Met residue (V173M), or the Vai residue is substituted for a Leu residue (V173L). In some embodiments, the p53 mutant peptide comprises a fragment of SEQ ID NO: 1 having an amino acid substitution at the 175^th amino acid from the N-terminus, which is an Arg residue. For example, the p53 mutant peptide may include an amino acid substitution at position 175 of SEQ ID NO: 1 wherein the Arg residue is substituted for a His residue (R175H). In some embodiments, the p53 mutant peptide comprises a fragment of SEQ ID NO: 1 having an amino acid substitution at the 176^th amino acid from the N-terminus, which is a Cys residue. For example, the p53 mutant peptide may include an amino acid substitution at position 176 of SEQ ID NO: 1 wherein the Cys residue is substituted for a Phe residue (C176F) or a Tyr residue (C176Y). In some embodiments, the p53 mutant peptide comprises a fragment of SEQ ID NO: 1 having an amino acid substitution at the 179^th amino acid from the N-terminus, which is a His residue. For example, the p53 mutant peptide may include an amino acid substitution at position 179 of SEQ ID NO: 1 wherein the His residue is substituted for a Tyr residue (H179Y) or an Arg residue (H179R). In some embodiments, the p53 mutant peptide comprises a fragment of SEQ ID NO: 1 having an amino acid substitution at the 193^rd amino acid from the N-terminus, which is a His residue. For example, the p53 mutant peptide may include an amino acid substitution at position 193 of SEQ ID NO: 1 wherein the His residue is substituted for an Arg residue (H193R). In some embodiments, the p53 mutant peptide comprises a fragment of SEQ ID NO: 1 having an amino acid substitution at the 195^th amino acid from the N-terminus, which is an lie residue. For example, the p53 mutant peptide may include an amino acid substitution at position 195 of SEQ ID NO: 1 wherein the lie residue is substituted for a Thr residue (I195T). In some embodiments, the p53 mutant peptide comprises a fragment of SEQ ID NO: 1 having an amino acid substitution at the 205^th amino acid from the N-terminus, which is a Tyr residue. For example, the p53 mutant peptide may include an amino acid substitution at position 205 of SEQ ID NO: 1 wherein the Tyr residue is substituted for a Cys residue (Y205C). In some embodiments, the p53 mutant peptide comprises a fragment of SEQ ID NO: 1 having an amino acid substitution at the 214^th amino acid from the N-terminus, which is a His residue. For example, the p53 mutant peptide may include an amino acid substitution at position 214 of SEQ ID NO: 1 wherein the His residue is substituted for an Arg residue (H214R). In some embodiments, the p53 mutant peptide comprises a fragment of SEQ ID NO: 1 having an amino acid substitution at the 216^th amino acid from the N-terminus, which is a Vai residue. For example, the p53 mutant peptide may include an amino acid substitution at position 216 of SEQ ID NO: 1 wherein the Vai residue is substituted for a Met residue (V216M). In some embodiments, the p53 mutant peptide comprises a fragment of SEQ ID NO: 1 having an amino acid substitution at the 220^th amino acid from the N-terminus, which is a Tyr residue. For example, the p53 mutant peptide may include an amino acid substitution at position 220 of SEQ ID NO: 1 wherein the Tyr residue is substituted for a Cys residue (Y220C). In some embodiments, the p53 mutant peptide comprises a fragment of SEQ ID NO: 1 having an amino acid substitution at the 234^th amino acid from the N-terminus, which is a Tyr residue. For example, the p53 mutant peptide may include an amino acid substitution at position 234 of SEQ ID NO: 1 wherein the Tyr residue is substituted for a Cys residue (Y234C). In some embodiments, the p53 mutant peptide comprises a fragment of SEQ ID NO: 1 having an amino acid substitution at the 236^th amino acid from the N-terminus, which is a Tyr residue. For example, the p53 mutant peptide may include an amino acid substitution at position 236 of SEQ ID NO: 1 wherein the Tyr residue is substituted for a Cys residue (Y236C). In some embodiments, the p53 mutant peptide comprises a fragment of SEQ ID NO: 1 having an amino acid substitution at the 237^th amino acid from the N-terminus, which is a Met residue. For example, the p53 mutant peptide may include an amino acid substitution at position 237 of SEQ ID NO: 1 wherein the Met residue is substituted for an lie residue (M237I). In some embodiments, the p53 mutant peptide comprises a fragment of SEQ ID NO: 1 having an amino acid substitution at the 238^th amino acid from the N-terminus, which is a Cys residue. For example, the p53 mutant peptide may include an amino acid substitution at position 238 of SEQ ID NO: 1 wherein the Cys residue is substituted for a Tyr residue (C238Y). In some embodiments, the p53 mutant peptide comprises a fragment of SEQ ID NO: 1 having an amino acid substitution at the 241 ^st amino acid from the N-terminus, which is a Ser residue. For example, the p53 mutant peptide may include an amino acid substitution at position 241 of SEQ ID NO: 1 wherein the Ser residue is substituted for a Phe residue (S241 F). In some embodiments, the p53 mutant peptide comprises a fragment of SEQ ID NO: 1 having an amino acid substitution at the 242^nd amino acid from the N-terminus, which is a Cys residue. For example, the p53 mutant peptide may include an amino acid substitution at position 242 of SEQ ID NO: 1 wherein the Cys residue is substituted for a Phe residue (C242F). In some embodiments, the p53 mutant peptide comprises a fragment of SEQ ID NO: 1 having an amino acid substitution at the 245^th amino acid from the N-terminus, which is a Gly residue. For example, the p53 mutant peptide may include an amino acid substitution at position 245 of SEQ ID NO: 1 wherein the Gly residue is substituted for a Cys residue (G245C), an Asp residue (G245D), a Ser residue (G245S), or a Vai residue (G245V). In some embodiments, the p53 mutant peptide comprises a fragment of SEQ ID NO: 1 having an amino acid substitution at the 248^th amino acid from the N-terminus, which is an Arg residue. For example, the p53 mutant peptide may include an amino acid substitution at position 248 of SEQ ID NO: 1 wherein the Arg residue is substituted for a Leu residue (R248L), a Gin residue (R248Q), or a Trp residue (R248W). In some embodiments, the p53 mutant peptide comprises a fragment of SEQ ID NO: 1 having an amino acid substitution at the 249^th amino acid from the N-terminus, which is an Arg residue. For example, the p53 mutant peptide may include an amino acid substitution at position 249 of SEQ ID NO: 1 wherein the Arg residue is substituted for a Ser residue (R249S). In some embodiments, the p53 mutant peptide comprises a fragment of SEQ ID NO: 1 having an amino acid substitution at the 266^th amino acid from the N-terminus, which is a Gly residue. For example, the p53 mutant peptide may include an amino acid substitution at position 266 of SEQ ID NO: 1 wherein the Gly residue is substituted for a Glu residue (G266E) or an Arg residue (G266R). In some embodiments, the p53 mutant peptide comprises a fragment of SEQ ID NO: 1 having an amino acid substitution at the 272^nd amino acid from the N-terminus, which is a Vai residue. For example, the p53 mutant peptide may include an amino acid substitution at position 272 of SEQ ID NO: 1 wherein the Vai residue is substituted for a Met residue (V272M). In some embodiments, the p53 mutant peptide comprises a fragment of SEQ ID NO: 1 having an amino acid substitution at the 273^rd amino acid from the N-terminus, which is an Arg residue. For example, the p53 mutant peptide may include an amino acid substitution at position 273 of SEQ ID NO: 1 wherein the Arg residue is substituted for a His residue (R273H), a Cys residue (R273C), or a Leu residue (R273L). In some embodiments, the p53 mutant peptide comprises a fragment of SEQ ID NO: 1 having an amino acid substitution at the 275^th amino acid from the N-terminus, which is a Cys residue. For example, the p53 mutant peptide may include an amino acid substitution at position 275 of SEQ ID NO: 1 wherein the Cys residue is substituted for a Tyr residue (C275Y). In some embodiments, the p53 mutant peptide comprises a fragment of SEQ ID NO: 1 having an amino acid substitution at the 278^th amino acid from the N-terminus, which is a Pro residue. For example, the p53 mutant peptide may include an amino acid substitution at position 278 of SEQ ID NO: 1 wherein the Pro residue is substituted for a Ser residue (P278S) or a Leu residue (P278L). In some embodiments, the p53 mutant peptide comprises a fragment of SEQ ID NO: 1 having an amino acid substitution at the 280^th amino acid from the N-terminus, which is an Arg residue. For example, the p53 mutant peptide may include an amino acid substitution at position 280 of SEQ ID NO: 1 wherein the Arg residue is substituted for a Thr residue (R280T) or a Lys residue (R280K). In some embodiments, the p53 mutant peptide comprises a fragment of SEQ ID NO: 1 having an amino acid substitution at the 282^th amino acid from the N-terminus, which is an Arg residue. For example, the p53 mutant peptide may include an amino acid substitution at position 282 of SEQ ID NO: 1 wherein the Arg residue is substituted for a Trp residue (R282W). In some embodiments, the p53 mutant peptide comprises a fragment of SEQ ID NO: 1 having an amino acid substitution at the 285^th amino acid from the N-terminus, which is a Glu residue. For example, the p53 mutant peptide may include an amino acid substitution at position 285 of SEQ ID NO: 1 wherein the Glu residue is substituted for a Lys residue (E285K). In some embodiments, the p53 mutant peptide comprises a fragment of SEQ ID NO: 1 having an amino acid substitution at the 286^th amino acid from the N-terminus, which is a Glu residue. For example, the p53 mutant peptide may include an amino acid substitution at position 286 of SEQ ID NO: 1 wherein the Glu residue is substituted for a Lys residue (E286K).

Table 1. Mutant p53 Peptides

In some embodiments, the peptide comprises an N-terminal modification. In some embodiments, the N-terminal modification is the addition of a cysteine. In some embodiments, the N-terminal modification is the addition of an acetylcysteine. In some embodiments, the N-terminal modification is the addition of a des-aminocysteine homolog. In some embodiments, the des-aminocysteine homolog is 3- mercaptopropionic acid or mercaptoacetic acid.

Amphiphilic Peptides

Amphiphilic peptides include a peptide and conjugated to an albumin-binding domain, e.g., a lipid. In some embodiments, the amphiphilic peptide includes a p53 or a mutant p53 peptide conjugated to an albumin-binding domain, e.g., a lipid, optionally by way of a linker. In some embodiments, the N- terminus of the peptide is bonded or linked to the albumin-binding domain. In some embodiments, the C- terminus of the peptide is bonded or linked to the albumin-binding domain. Lipids

The compounds described herein include herein a p53 peptide or a p53 mutant peptide that are conjugated to an albumin-binding domain. In some embodiments, the albumin-binding domain is a lipid. The lipid can be linear, branched, or cyclic.

Examples of preferred lipids include, but are not limited to, fatty acids with aliphatic tails of 3-30 carbons including, but not limited to, linear unsaturated and saturated fatty acids, branched saturated and unsaturated fatty acids, and fatty acids derivatives, such as fatty acid esters, fatty acid amides, and fatty acid thioesters, diacyl lipids, cholesterol, cholesterol derivatives, and steroid acids such as bile acids, Lipid A or combinations thereof.

In certain embodiments, the lipid is a diacyl lipid or two-tailed lipid. In some embodiments, the tails in the diacyl lipid contain from about 12 to about 30 carbons (e.g., 13 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, or 29). In some embodiments the tails in the diacyl lipid contain about 14 to about 25 carbons (e.g., 15, 16, 17, 18, 19, 20, 21 , 22, 23, or 24). In some embodiments, the tails of the diacyl lipid contain from about 16 to about 20 carbons (e.g., 17, 18, or 19). In some embodiments, the diacyl lipid comprises 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29 or 30 carbons.

The carbon tails of the diacyl lipid can be saturated, unsaturated, or combinations thereof. The tails can be coupled to the head group via ester bond linkages, amide bond linkages, thioester bond linkages, or combinations thereof. In a particular embodiment, the diacyl lipids are phosphate lipids, glycolipids, sphingolipids, or combinations thereof.

In some embodiments, the lipid is 1 ,2-distearoyi-s/7-giycero-3-phosphoethanolamine (DSPE).

In some embodiments, the p53 peptide or mutant p53 peptide bonded or linked by a linker to the following lipid:

or a salt thereof, wherein X is O or S.

The p53 peptide or mutant p53 peptide may be directly bonded to the lipid. Alternatively, the p53 peptide or mutant p53 peptide may be linked to the lipid through a linker.

Reference to lipids herein, as well as amphiphiles including the lipid, is to be understood as including pharmaceutically acceptable salts thereof.

Linkers

In some embodiments, the compound includes a p53 peptide or a mutant p53 peptide linked to an albumin-binding domain, e.g., a lipid, by a linker. The linker may be a hydrophilic polymer, a string of hydrophilic amino acids, a polysaccharide, and an oligonucleotide, or a combination thereof. The linker may reduce or prevent the ability of the albumin-binding domain to insert into the plasma membrane of cells, such as cells in the tissue adjacent to the injection site. The linker can also reduce or prevent the ability of the amphiphilic peptide sequence from non-specifically associating with extracellular matrix proteins at the site of administration. For the amphiphilic p53 peptide or mutant p53 peptide to be trafficked efficiently to the lymph node, it should remain soluble. A polar block linker may be included between the p53 peptide or mutant p53 peptide and the albumin-binding domain to which it is conjugated to increase solubility of the amphiphilic p53 peptide or mutant p53 peptide.

The length and composition of the linker can be adjusted based on the albumin-binding domain and the peptide selected. For example, in certain embodiments, the polynucleotide itself may be polar enough to ensure solubility; for example, polynucleotides that are 10, 15, 20 or more nucleotides in length. Therefore, in some embodiments, no additional linker is required. However, in certain cases, it can be desirable to include a linker that mimics the effect of a polar oligonucleotide. A linker can be used as part of any of albumin-binding domain conjugates described herein, for example, lipid-oligonucleotide conjugates and lipid-peptide conjugates, which reduce cell membrane insertion/preferential portioning onto albumin.

Suitable linkers include, but are not limited to, oligonucleotides such as those discussed above, including a string of nucleic acids, a hydrophilic polymer including but not limited to polyethylene glycol) (MW: 500 Da to 20,000 Da), polyacrylamide (MW: 500 Da to 20,000 Da), polyacrylic acid; a string of hydrophilic amino acids such as serine, threonine, cysteine, tyrosine, asparagine, glutamine, aspartic acid, glutamic acid, lysine, arginine, histidine, or combinations thereof; polysaccharides, including but not limited to, dextran (MW: 1 ,000 Da to 2,000,000 Da), or combinations thereof. The hydrophobic albuminbinding domain and the linker/peptide (e.g., the p53 peptide or mutant p53 peptide) are covalently linked. The covalent bond may be a non-cleavable linkage or a cleavable linkage. The non-cleavable linkage can include an amide bond or phosphate bond, and the cleavable linkage can include a disulfide bond, acid- cleavable linkage, ester bond, anhydride bond, biodegradable bond, or enzyme-cleavable linkage.

In some embodiments, the linker is one or more ethylene glycol (EG) units, more preferably two or more EG units (i.e., polyethylene glycol (PEG)). For example, in some embodiments, the compound includes a p53 peptide or a mutant p53 peptide and a hydrophobic albumin-binding domain linked by a polyethylene glycol (PEG) molecule or a derivative or analog thereof.

In some embodiments, compounds described herein contain a p53 peptide or a mutant p53 peptide linked to PEG which is in turn linked to a hydrophobic albumin-binding domain, e.g., a lipid. The precise number of PEG units depends on the albumin-binding domain and the cargo, however, typically, a linker can have between about 1 and about 100 (e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 35, 36, 37, 38, 39, 40, 41 , 42, 43, 44,

45, 46, 47, 48, 49, 50, 51 , 52, 53, 54, 55, 56, 57, 58, 59, 60, 61 , 62, 63, 64, 65, 66, 67, 68, 69, 70, 71 , 72,

73, 74, 75, 76, 77, 78, 79, 80, 81 , 82, 83, 84, 85, 86, 87, 88, 89, 90, 91 , 92, 93, 94, 95, 96, 97, 98, 99, or

100). In some embodiments, the linker may be a PEG linker having between about 20 and about 80, between about 30 and about 70, or between about 40 and about 60 PEG units. In some embodiments, the number of PEG units is between 24 and 50 units (e.g., between 24 and 45, 24 and 40, 24 and 35, 24 and 30, 30 and 50, 35 and 50, 40 and 50, and 45 and 50 units). In some embodiments, the linker has between about 45 and 55 PEG units. For example, in some embodiments, the linker has 48 PEG units. In some embodiments, the linker includes a PEG4-amido-PEG4 linker.

As discussed above, in some embodiments, the linker is an oligonucleotide which includes a string of nucleic acids. In some embodiments, the compounds described herein include a p53 peptide or a mutant p53 peptide linked to a string of nucleic acids, which is in turn linked to a hydrophobic albumin- binding domain, e.g., a lipid. The linker can be any sequence, for example, the sequence of the oligonucleotide can be a random sequence, or a sequence specifically chosen for its molecular or biochemical properties (e.g., highly polar). In some embodiments, the linker includes 20 one or more series of consecutive adenine (A), cytosine (C), guanine (G), thymine (T), uracil (U), or analog thereof. In some embodiments, the linker consists of a series of consecutive adenine (A), cytosine (C), guanine (G), thymine (T), uracil (U), or analog thereof.

In some embodiments, the string of nucleic acids includes between 1 and 50 nucleic acid residues. In some embodiments, the string of nucleic acids includes between 5 and 30 nucleic acid residues. In some embodiments, the linker includes one or more guanines, for example between 1 -10 guanines.

In some embodiments, the linker is an oligonucleotide that includes a string of amino acids. In some embodiments, the amphiphilic p53 peptide or mutant p53 peptide which is in turn linked to a hydrophobic albumin-binding domain, e.g., a lipid. The linker can have any amino acid sequence, for example, the sequence of the oligonucleotide can be a random sequence, or a sequence chosen for its molecular or biochemical properties (e.g., high flexibility). In some embodiments, the linker includes a series of glycine residue to form a polyglycine linker. In some embodiments, the linker includes an amino acid sequence of (Gly)n, wherein n may be between 2 and 20 residues. Examples of polyglycine linkers include but are not limited to GGG, GGGA (SEQ ID NO: 7), GGGG (SEQ ID NO: 8), GGGAG (SEQ ID NO: 9), GGGAGG (SEQ ID NO: 10), GGGAGGG (SEQ ID NO: 11 ), GGAG (SEQ ID NO: 12),GGSG (SEQ ID NO: 13), AGGG (SEQ ID NO: 14), SGGG (SEQ ID NO: 15), GGAGGA (SEQ ID NO: 16), GGSGGS (SEQ ID NO: 17), GGAGGAGGA (SEQ ID NO: 18), GGSGGSGGS (SEQ ID NO: 19), GGAGGAGGAGGA (SEQ ID NO: 20), GGSGGSGGSGGS (SEQ ID NO: 21 ), GGAGGGAG (SEQ ID NO: 22), GGSGGGSG (SEQ ID NO: 23), GGAGGGAGGGAG (SEQ ID NO: 24), GGSGGGSGGGSG (SEQ ID NO: 25), GGGGAGGGGAGGGGA (SEQ ID NO: 26), and GGGGSGGGGSGGGGS (SEQ ID NO: 27).

Methods of Conjugation

Described herein are compounds including a p53 peptide or a mutant p53 peptide and an albumin-binding domain.

The peptide may be modified with N-terminal cysteine, acetyl-cysteine, sulfydryl, transcyclooctene, cyclooctyne, azide or alkyne for the conjugation with a mutant p53 peptide and an albuminbinding domain. In some embodiments, the peptide is modified with C-terminal cysteine, azide or alkyne for the conjugation with a p53 peptide or a mutant p53 peptide and an albumin-binding domain. In some embodiments, the internal cysteine or lysine of a peptide is used for the conjugation with an albuminbinding domain.

The p53 peptide or mutant p53 peptide and the albumin-binding domain may be bonded or linked to a linker. In some embodiments, the linker includes a functional group. In some embodiments, the functional group is capable of conjugating to a peptide. For example, the p53 peptide or mutant p53 peptide may be bound to a linker, wherein the linker is modified with a functional group. In some embodiments, the albumin-binding domain may be linked to a linker, wherein the linker is modified with a functional group. In some embodiments, the linker may be a PEG linker. In some embodiments, the p53 peptide or mutant p53 peptide is conjugated to the albuminbinding domain and/or linker by way of a reaction between a dithio group and a free thiol group.

Adjuvants

In some embodiments, a pharmaceutical composition described herein may be administered with one or more adjuvants. An adjuvant refers to a substance that cause stimulation of the immune system. In this context, an adjuvant is used to enhance an immune response to the peptide. An adjuvant may be administered to a subject before, in combination with, or after administration of the compositions described herein. In some embodiments, an additional adjuvant is administered to the subject in combination with the p53 peptide or mutant p53 peptide conjugated to an albumin-binding domain described herein. In some embodiments, an adjuvant may be conjugated to an albumin-binding domain, e.g., a lipid. The adjuvant may be without limitation lipids (e.g., monophosphoryl lipid A (MPLA)), alum (e.g., aluminum hydroxide, aluminum phosphate); Freund’s adjuvant; saponins purified from the bark of the Q. saponaria tree such as QS21 (a glycolipid that elutes in the 21st peak with HPLC fractionation; Antigenics, Inc., Worcester, Mass.); poly[di(carboxylatophenoxy)phosphazene (PCPP polymer; Virus Research Institute, USA), Flt3 ligand, Leishmania elongation factor (a purified Leishmania protein; Corixa Corporation, Seattle, Wash.), ISCOMS (immunostimulating complexes which contain mixed saponins, lipids and form virus-sized particles with pores that can hold an antigen; CSL, Melbourne, Australia), Pam3Cys, SB-AS4 (SmithKline Beecham adjuvant system #4 which contains alum and MPL; SBB, Belgium), non-ionic block copolymers that form micelles such as CRL 1005 (these contain a linear chain of hydrophobic polyoxypropylene flanked by chains of polyoxyethylene, Vaxcel, Inc., Norcross, Ga.), and Montanide IMS (e.g., IMS1312, water-based nanoparticles combined with a soluble immunostimulant, Seppic), and CDNs (cyclic di-nucleotides).

Adjuvants may be toll-like receptor (TLR) ligands. Adjuvants that act through TLR3 include without limitation double-stranded RNA. Adjuvants that act through TLR4 include without limitation derivatives of lipopolysaccharides such as monophosphoryl lipid A (MPLA; Ribi ImmunoChem Research, Inc., Hamilton, Mont.) and muramyl dipeptide (MDP; Ribi) andthreonyl-muramyl dipeptide (t-MDP; Ribi); OM-174 (a glucosamine disaccharide related to lipid A; OM Pharma SA, Meyrin, Switzerland). Adjuvants that act through TLR5 include without limitation flagellin. Adjuvants that act through TLR7 and/or TLR8 include single-stranded RNA, oligoribonucleotides (ORN), synthetic low molecular weight compounds such as imidazoquinolinamines (e.g., imiquimod (R-837), resiquimod (R-848)). Adjuvants 5 acting through TLR9 include DNA of viral or bacterial origin, or synthetic oligodeoxynucleotides (ODN), such as CpG ODN. For example, the CpG ODN may have a sequence of TCGTCGTTTTGTCGTTTTGTCGTT-3’ (SEQ ID NO: 4), 5’-TGACTGTGAACGTTCGAGATGA-3’ (SEQ ID NO: 5), or 5’- TCGTCGTTTTCGGCGCGCGCCG-3’ (SEQ ID NO: 6). The linkages of the CpG may be all phosphorothioate linkages.

Another adjuvant class is phosphorothioate containing molecules such as phosphorothioate nucleotide analogs and nucleic acids containing phosphorothioate backbone linkages. Pharmaceutical Compositions

Described herein are pharmaceutical compositions of the disclosure including a p53 peptide or a mutant p53 peptide conjugated to an albumin-binding domain. In addition to a therapeutic amount of the p53 peptide or mutant p53 peptide conjugated to an albumin-binding domain described herein, the pharmaceutical compositions may contain a pharmaceutically acceptable carrier or excipient, which can be formulated by methods known to those skilled in the art. Pharmaceutically acceptable salts of the components are also included, as described herein.

Acceptable carriers and excipients in the pharmaceutical compositions of the p53 peptide or mutant p53 peptide conjugated to an albumin-binding domain described herein are nontoxic to recipients at the dosages and concentrations employed. In certain embodiments, the formulation material(s) are for subcutaneous (s.c.), intramuscular (i.m.), and/or intravenous (i.v.) administration. In some embodiments, administration is by inhalation or intranasal administration. In some embodiments, the formulation material(s) intraperitoneal, topical, or oral administration. In some embodiments, the pharmaceutical composition can contain formulation materials for modifying, maintaining or preserving, for example, the pH, osmolality, viscosity, clarity, color, isotonicity, odor, sterility, stability, rate of dissolution or release, adsorption or penetration of the composition. In some embodiments, suitable formulation materials include, but are not limited to, amino acids (such as glycine, glutamine, asparagine, arginine or lysine); antimicrobials; antioxidants (such as ascorbic acid, methionine, sodium sulfite or sodium hydrogensulfite); buffers (such as borate, bicarbonate, Tris-HCI, citrates, HEPES, TAE, phosphates or other organic acids); bulking agents (such as mannitol or glycine); chelating agents (such as ethylenediamine tetraacetic acid (EDTA)); complexing agents (such as caffeine, polyvinylpyrrolidone, beta-cyclodextrin or hydroxypropyl-beta- cyclodextrin); fillers; monosaccharides; disaccharides; and other carbohydrates (such as glucose, sucrose, mannose or dextran); proteins (such as human serum albumin, gelatin, dextran, and immunoglobulins); coloring, flavoring and diluting agents; emulsifying agents; hydrophilic polymers (such as polyvinylpyrrolidone); low molecular weight polypeptides; salt-forming counterions (such as sodium); preservatives (such as hexamethonium chloride, octadecyldimethylbenzyl ammonium chloride, resorcinol, and benzalkonium chloride, benzoic acid, salicylic acid, thimerosal, phenethyl alcohol, methylparaben, propylparaben, chlorhexidine, sorbic acid or hydrogen peroxide); solvents (such as glycerin, propylene glycol or polyethylene glycol); sugar alcohols (such as mannitol or sorbitol); suspending agents; surfactants or wetting agents (such as pluronics, PEG, sorbitan esters, polysorbates such as polysorbate 20, polysorbate 80, triton, tromethamine, lecithin, cholesterol, tyloxapal); stability enhancing agents (such as sucrose or sorbitol); tonicity enhancing agents (such as alkali metal halides, preferably sodium or potassium chloride, mannitol sorbitol); delivery vehicles; diluents; excipients and/or pharmaceutical adjuvants. (Remington's Pharmaceutical Sciences, 18th Edition, A. R. Gennaro, ed., Mack Publishing Company (1995). In some embodiments, the optimal pharmaceutical composition will be determined by one skilled in the art depending upon, for example, the intended route of administration, delivery format and desired dosage. See, for example, Remington's Pharmaceutical Sciences, supra. In some embodiments, such compositions may influence the physical state, stability, rate of in vivo release and rate of in vivo clearance of the amphiphilic conjugate.

In some embodiments, the primary vehicle or carrier in a pharmaceutical composition, including the p53 peptide or mutant p53 peptide conjugated to an albumin-binding domain described herein can be either aqueous or non-aqueous in nature. For example, in some embodiments, a suitable vehicle or carrier can be water for injection, physiological saline solution, or artificial cerebrospinal fluid, possibly supplemented with other materials common in compositions for parenteral administration. In some embodiments, the saline includes isotonic phosphate-buffered saline. In certain embodiments, neutral buffered saline or saline mixed with serum albumin are further exemplary vehicles. In some embodiments, pharmaceutical compositions include Tris buffer of about pH 7.0-8.5, or acetate buffer of about pH 4.0-5.5, which can further include sorbitol or a suitable substitute therefore. In some embodiments, a composition including the p53 peptide or mutant p53 peptide conjugated to an albuminbinding domain described herein can be prepared for storage by mixing the selected composition having the desired degree of purity with optional formulation agents (Remington's Pharmaceutical Sciences, supra) in the form of a lyophilized cake or an aqueous solution. Further, in some embodiments, the composition including the p53 peptide or mutant p53 peptide conjugated to an albumin-binding domain described herein can be formulated as a lyophilizate using appropriate excipients such as sucrose.

In some embodiments, the pharmaceutical composition may be selected for parenteral delivery. The preparation of such pharmaceutically acceptable compositions is within the ability of one skilled in the art.

In some embodiments, the formulation components are present in concentrations that are acceptable to the site of administration. In some embodiments, buffers are used to maintain the composition at physiological pH or at a slightly lower pH, typically within a pH range of from about 5 to about 8.

In some embodiments, when parenteral administration is contemplated, a therapeutic composition can be in the form of a pyrogen-free, parenterally acceptable aqueous solution including the p53 peptide or mutant p53 peptide conjugated to an albumin-binding domain described herein in a pharmaceutically acceptable vehicle. In some embodiments, a vehicle for parenteral injection is sterile distilled water in which a p53 peptide or a mutant p53 peptide conjugated to an albumin-binding domain described herein is formulated as a sterile, isotonic solution, properly preserved. In some embodiments, the preparation can involve the formulation of the desired molecule with an agent, such as injectable microspheres, bio-erodible particles, polymeric compounds (such as polylactic acid or polyglycolic acid), beads or liposomes, that can provide for the controlled or sustained release of the product which can then be delivered via a depot injection. In some embodiments, hyaluronic acid can also be used, and can have the effect of promoting sustained duration in the circulation. In some embodiments, implantable drug delivery devices can be used to introduce the desired molecule.

The pharmaceutical composition may be administered in therapeutically effective amount such as to induce an immune response. The therapeutically effective amount of the p53 peptide or mutant p53 peptide conjugated to an albumin-binding domain described herein included in the pharmaceutical preparations may be determined by one of skill in art, such that the dosage (e.g., a dose within the range of 0.01 -100 mg/kg of body weight) induces an immune response in the subject.

Vectors may be used as in vivo nucleic acid delivery vehicle include, but are not limited to, retroviral vectors, adenoviral vectors, poxviral vectors (e.g., vaccinia viral vectors, such as Modified Vaccinia Ankara (MVA)), adeno-associated viral vectors, and alphaviral vectors. In some embodiments, a vector can include internal ribosome entry site (IRES) that allows the expression of peptides described herein. Other vehicles and methods for nucleic acid delivery are described in US Patent Nos. 5,972,707, 5,697,901 , and 6,261 ,554, each of which is incorporated by reference herein in its entirety. Other methods of producing pharmaceutical compositions are described in, e.g., US Patent Nos. 5,478,925, 8,603,778, 7,662,367, and 7,892,558, all of which are incorporated by reference herein in their entireties.

In some embodiments, a pharmaceutical composition described herein may be administered with one or more adjuvants.

Routes, Dosage, and Timing of Administration

Pharmaceutical compositions of the disclosure that contain the p53 peptide or mutant p53 peptide conjugated to an albumin-binding domain described herein as the therapeutic agent may be formulated for parenteral administration, subcutaneous administration, intravenous administration, intramuscular administration, intranasal administration, or inhalation. In some embodiments, the therapeutic agent is formulated for transmucosal administration. In some embodiments, the therapeutic agent is formulated for buccal administration. In some embodiments, the therapeutic agent is formulated for sublingual administration. Methods of administering therapeutic proteins are known in the art. See, for example, US Patent Nos. 6,174,529, 6,613,332, 8,518,869, 7,402,155, and 6,591 ,129, and US Patent Application Publication Nos. US20140051634, WO1993000077, and US20110184145, the disclosures of which are incorporated by reference in their entireties.

One or more of these methods may be used to administer a pharmaceutical composition of the invention that contains a p53 peptide or mutant p53 peptide conjugated to an albumin-binding domain. For injectable formulations, various effective pharmaceutical carriers are known in the art. See, e.g., Pharmaceutics and Pharmacy Practice, J. B. Lippincott Company, Philadelphia, Pa., Banker and Chalmers, eds., pages 238-250 (1982), and ASHP Handbook on Injectable Drugs, Toissel, 4th ed., pages 622-630 (1986). The dosage of the pharmaceutical compositions of the invention depends on factors including the route of administration and the physical characteristics, e.g., age, weight, general health, of the subject. Typically, the amount of a p53 peptide or mutant p53 peptide conjugated to an albuminbinding domain described herein contained within a single dose may be an amount that effectively induces an immune response in the subject without inducing significant toxicity. A pharmaceutical composition of the invention may include a dosage of a p53 peptide or a mutant p53 peptide conjugated to an albumin-binding domain described herein ranging from 0.001 to 500 mg (e.g., 0.01 , 0.05, 0.1 , 0.2, 0.3, 0.5, 0.7, 0.8, 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 10 mg, 15 mg, 20 mg, 30 mg, 50 mg, 100 mg, 250 mg, or 500 mg) and, in a more specific embodiment, about 0.1 to about 100 mg. The dosage may be adapted by the clinician in accordance with the different parameters of the subject.

Pharmaceutical compositions of the invention that contain a p53 peptide or a mutant p53 peptide conjugated to an albumin-binding domain may be administered to a subject in need thereof, for example, one or more times (e.g., 1 -10 times or more) daily, weekly, monthly, biannually, annually, or as medically necessary. Methods of Inducing an Immune Response

The disclosure provides methods of inducing an immune response against the p53 peptide or mutant p53 peptide in a subject. The method includes administering any one of the compounds described herein to the subject.

In some embodiments, the disclosure provides a method of inducing an immune response against the p53 peptide or mutant p53 peptide in subject by administering any one of the p53 peptide or mutant p53 peptide conjugated to an albumin-binding domain to the subject and further administering an adjuvant to the subject. In some embodiments, the p53 peptide or mutant p53 peptide conjugated to an albumin-binding domain may be administered without one or more additional adjuvants.

In some embodiments, the method includes administering to the subject a therapeutically effective amount of the p53 peptide or mutant p53 peptide conjugated to an albumin-binding domain described herein. In some embodiments, the p53 peptide or mutant p53 peptide conjugated to an albumin-binding domain is administered substantially simultaneously. In some embodiments, the p53 peptide or mutant p53 peptide conjugated to an albumin-binding domain is administered separately.

In some embodiments, one or more of the components administered is a pharmaceutically acceptable salt of the indicated component, as described herein.

In some embodiments, the disclosure provides a method of inducing an immune response against the peptide in a subject by administering any one of the compounds or pharmaceutically acceptable salts described herein subcutaneously to the subject. In some embodiments, the disclosure provides a method of inducing an immune response against the peptide in a subject by administering the peptide intramuscularly, subcutaneously, intravenously, intraperitoneally, topically, orally/buccally, sublingually, transmucosally, intranasally, or by inhalation to the subject.

In some embodiments, the subject is a mammal. For example, the subject may be a human.

Kits

A kit can include the p53 peptide or mutant p53 peptide conjugated to an albumin-binding domains disclosed herein and instructions for use. The kits may include, in a suitable container, a p53 peptide or a mutant p53 peptide conjugated to an albumin-binding domain, one or more controls, and various buffers, reagents, enzymes and other standard ingredients well known in the art. In some embodiments, the kits further include an adjuvant. Accordingly, in some embodiments, the p53 peptide or mutant p53 peptide conjugated to an albumin-binding domain is in a vial. In some embodiments, the p53 peptide or mutant p53 peptide conjugated to an albumin-binding domain and the adjuvant are in separate vials. In some embodiments, the peptide and adjuvant are in the same vial. In some embodiments, the peptide and the adjuvant are in separate vials.

The container can include at least one vial, well, test tube, flask, bottle, syringe, or other container means, into which the p53 peptide or mutant p53 peptide conjugated to an albumin-binding domain and in some instances, suitably aliquoted. When an additional component is provided, the kit can contain additional containers into which this compound may be placed. The kits can also include a means for containing the p53 peptide or mutant p53 peptide conjugated to an albumin-binding domain and any other reagent containers in close confinement for commercial sale. Such containers may include injection or blow-molded plastic containers into which the desired vials are retained. Containers and/or kits can include labeling with instructions for use and/or warnings.

In some embodiments, the disclosure provides a kit including a medicament including a composition including a p53 peptide or a mutant p53 peptide conjugated to an albumin-binding domain, an optional pharmaceutically acceptable carrier, and a package insert including instructions for administration of the medicament alone or in combination with a composition including an adjuvant and an optional pharmaceutically acceptable carrier, for treating, delaying progression of, or preventing a disease or condition.

In some embodiments, the disclosure provides a kit including a container including a composition including a p53 peptide or a mutant p53 peptide conjugated to an albumin-binding domain, an optional pharmaceutically acceptable carrier, and a package insert including instructions for administration of composition vaccine in a subject, wherein the p53 peptide or mutant p53 peptide is conjugated to an albumin-binding domain and optionally includes a linker. In some embodiments, the kit further includes an adjuvant and instructions for administration of the adjuvant.

In some embodiments of the kits, one of more of the components of the kits is a pharmaceutically acceptable salt of the component as described herein.

EXAMPLES

The following examples, which are intended to illustrate, rather than limit, the disclosure, are put forth to provide those of ordinary skill in the art with a description of how the compositions and methods described herein may be used, made, and evaluated. The examples are intended to be purely exemplary of the disclosure and are not intended to limit the scope of what the inventors regard as their invention.

Example 1. Ability of p53 R248W peptides to elicit an immune response

The purpose of this experiment was to establish if an immune response could be elicited against p53 R248W with AMP-conjugated p53 peptides (801/802) and AMP-CpG7909.

5 groups of 10 C57BL/6J mice each were administered a vaccine including the components of Table 3. 5 mice in each group were taken down after two doses and the remaining 5 mice were taken down after three doses as shown in Table 2.

Table 2. Summary of Vaccine Administration in Mice

The amount of peptide-antigen used was 5 nmol per injection. Because of low solubility of some AMP-peptides, all AMP-peptide stocks were co-solubilized with AMP-CpG. To achieve this, lyophilized AMP-peptides were dissolved in 50% t-butanol first, followed by the addition of 1 :1 molar ratio of AMP- CpG and subsequent lyophilization. The obtained powder was then resuspended in 1 X PBS to a concentration of 1 mg/ml. The vaccine components are described in Table 3. AMP-vaccine stocks were further diluted to their final concentrations using 1 X PBS such that each injection contained 5 nmol AMP- antigen and 5 nmol AMP-adjuvant.

The soluble peptide stock solutions were prepared in either 1 .1 X PBS (802) or water (801 ), due to their solubility, at a concentration of 1 mg/ml and 0.25 mg/ml, respectively and further diluted with 1 X PBS such that the final concentration of soluble peptide was 5 nmol/100 pL injection. The soluble adjuvant stock solutions (CpG) were prepared in limulus amebocyte lysate (LAL) H2O and further diluted with 1 X PBS such that the final concentration was 5 nmol/100 pL injection.

The immunizations were administered subcutaneously (SC) into the tail base of female B6 mice, bilaterally with 50 pL per side. A booster dose was given at roughly 2-week intervals. SC injections ensured that the vaccine was optimally delivered into lymph nodes via natural lymph drainage, and biweekly injections were determined to be optimal in immune response.

Table 3. Vaccine Components

NMS - N-Methylsuccinimide

ELISpot analysis for IFNy was performed on splenocytes 7 days after dose 2, with 0.1 x10⁶ cells/well and 2 pg/ml of each peptide, and after dose 3, with 1 x10⁶ cells/well and 20 pg/ml of each peptide. Splenocytes were activated with the p53 peptides described in Table 4. IFNy plates were stimulated overnight. The results of this analysis are shown in FIGS. 2A and 2B.

Table 4. Re-Stimulation Peptides

Immunization with AMP-conjugated, but not with soluble, p53 R248W 10-mer and 18-mer elicited a significant immune response in mice after dose 2, which was further potentiated after administration of a third dose.

Example 2. Ability of p53 R248Q peptides to elicit an immune response

This experiment aimed to determine if an immune response could be elicited against p53 R248Q using AMP-conjugated p53 peptides and AMP-CpG7909.

5 groups of 10 C57BL/6J mice each were administered a vaccine including the components of Table 6. 5 mice in each group were taken down after two doses and the remaining 5 mice were taken down after three doses as shown in Table 5.

Table 5. Summary of Vaccine Administration in Mice

The amount of peptide-antigen used was 5 nmol per injection. Because of low solubility of some AMP-peptides, all AMP-peptide stocks were co-solubilized with AMP-CpG. To achieve this, lyophilized AMP-peptides were dissolved in 50% t-butanol first, followed by the addition of 1 :1 molar ratio of AMP- CpG and subsequent lyophilization. The obtained powder was then resuspended in 1X PBS to a concentration of 1 mg/ml. The vaccine components are described in Table 6. AMP-vaccine stocks were further diluted to their final concentrations using 1 X PBS such that each injection contained 5 nmol AMP- antigen and 5 nmol AMP-adjuvant.

Soluble peptide stock solutions were prepared in 1 .1 X PBS at a concentration of 0.5 mg/mL and further diluted with 1 X PBS such that the final concentration was 5 nmol/100 pL injection. The soluble adjuvant stock solutions (CpG) were prepared in limulus amebocyte lysate (LAL) H2O and further diluted with 1 X PBS such that the final concentration was 5 nmol/100 pL injection.

Table 6. Vaccine Components

NMS - N-Methylsuccinimide

ELISpot analysis for IFNy was performed on splenocytes 7 days after dose 2, with 0.1 x10⁶ cells/well and 2 pg/ml of each peptide, and after dose 3, with 1 x10⁶ cells/well and 20 pg/ml of each peptide. Splenocytes were activated with the p53 peptides described in Table 7. IFNy plates were stimulated overnight. The results of this analysis are shown in FIGS. 3A and 3B.

Table 7. Re-Stimulation Peptides

Immunization with AMP-conjugated, but not with soluble, p53 R248Q 10-mer and 18-mer elicited a detectable immune response in mice.

Example 3. Effect of dosage of p53 R248W peptides on immune response

This experiment aimed to establish the optimal dose for p53 R248W peptides when administered subcutaneously according to several candidate immunization regimens.

9 groups of 5 C57BL/6J mice each were administered a vaccine including the components of Table 9. 5 mice in each group were taken down after three doses as shown in Table 8. Table 8. Summary of Vaccine Administration in Mice

The amount of peptide-antigen used was a range of doses from 1 to 20 nmol peptide per injection. Because of low solubility of some AMP-peptides, all AMP-peptide stocks were co-solubilized with AMP-CpG. To achieve this, AMP-peptides were dissolved in 50% t-butanol first, followed by the addition of 1 :4, 1 :1 or 2:1 molar ratio of AMP-CpG and subsequent lyophilization. The obtained powder was then resuspended in 1 X PBS to a concentration of close to 1 mg/ml. The vaccine components are described in Table 9. AMP-vaccine stocks were further diluted to final concentrations using 1X PBS. AMP-vaccines contained 1 , 5, 10 or 20 nmol AMP-antigen and 5 or 10 nmol AMP-adjuvant (Table 8).

The immunizations were administered subcutaneously (SC) into the tail base of female B6 mice, bilaterally with 50 pL per side. 2 mice received only half a dose during Dose 1 (on right side): Group 3 mouse 5, and Group 6 mouse 5. SC injections ensured that the vaccine was optimally delivered into lymph nodes via natural lymph drainage, and bi-weekly injections were determined to be optimal in immune response.

Table 9. Vaccine Components

An Intracellular Stain (ICS) assay to assess TNFa and IFNy levels was performed on CD4⁺ and CD8⁺ T cells from spleen (1 x10⁶ cells/well) 7 days after dose 3, as shown in FIG. 6A and FIG. 6B, respectively. Cells were also stained for CD4, CD8, and CD3 using the antibodies described in Table 10. ICS samples were activated overnight (in the presence of Brefeldin A and Monensin) with 2 pg/ml of each peptide listed in Table 11 .

Table 10. Antibodies used for ICS

Table 11. Re-Stimulation Peptides

ELISpot analysis for IFNy was performed on splenocytes 7 days after dose 3, with 0.2x10⁶ cells/well and 2 pg/ml of each peptide. Splenocytes were activated with the p53 peptides described in Table 11 . IFNy plates were stimulated overnight. The results of this analysis are shown in FIG. 4.

Luminex analysis was conducted on splenocytes 7 days after dose 3. 1 x10⁶ cells/well were activated with 2 pg/ml of each p53 peptide (Table 11 ). Supernatant was collected, and tested with mouse cytokine/chemokine magnetic bead kit (See Error! Reference source not found.12) for the simultaneous quantification of the following 5 analytes: GM-CSF, IL2, TNFa, IN Fy, and Granzyme B, as shown in FIGS. 5A-5E.

Table 12. Luminex kit

Granzyme B Millipore Sigma MCD8MAG-48K-01

Example 4. Dosing schedules of p53 R248W peptides

This experiment aimed to determine the optimal dosing schedule for future mouse studies using the R248W peptides.

6 groups of 10 C57BL/6J mice in Groups 2-5 and 5 C57BL/6J mice in Groups 1 and 6 each were administered a vaccine including the components of Table 14. Groups were dosed at different times culminating in a common take-down date for all groups as described in Table 13.

Table 13. Summary of Vaccine Administration in Mice

The optimal amount of peptide-antigen was determined by previous experiments to be 1 .25 nmol of peptide per injection. Because of low solubility of some AMP-peptides, all AMP-peptide stocks were co-solubilized with AMP-CpG. To achieve this, AMP-peptides were dissolved in 50% t-butanol first, followed by the addition of 1 :4 molar ratio of AMP-CpG and subsequent lyophilization. The obtained powder was then resuspended in 1X PBS to a concentration of close to 1 mg/ml. The vaccine components are described in Table 14. AMP-vaccine stocks were further diluted to final concentrations using 1X PBS. AMP-vaccines contained 1 .25 nmol AMP-antigen and 5 nmol AMP-adjuvant.

Soluble peptides were prepared in 1 .1 X PBS at a concentration of 1 mg/ml and further diluted with 1 X PBS such that the final concentration of the soluble peptide was 1 .25 nmol/100 pL injection. The soluble adjuvant solutions (CpG) were prepared in limulus amebocyte lysate (LAL) H2O and further diluted with 1 X PBS such that the final concentration was 5 nmol/100 pL injection.

The immunizations were administered subcutaneously (SC) into the tail base of female B6 mice, bilaterally with 50 pL per side. SC injections ensured that the vaccine was optimally delivered into lymph nodes via natural lymph drainage, and bi-weekly injections were determined to be optimal in immune response. Table 14. Vaccine Components

An ICS assay to assess TNFa and IFNy levels was performed on splenocytes (1 x10⁶ cells/well) on day 35, as shown in FIG. 9. Cells were also stained for CD4, CD8, and CD3 using the antibodies described in Table 15. ICS samples were activated overnight (in the presence of Brefeldin A and Monensin) with 2 pg/ml of each peptide listed in Table 16.

Table 15. Antibodies used for ICS

Table 16. Re-Stimulation Peptides

ELISpot analysis for IFNy was performed on splenocytes on day 35 with 0.2x10⁶ cells/well and 2 pg/ml of each peptide, as shown in FIG. 7. Splenocytes were activated with the p53 peptides described in Table 16. IFNy plates were stimulated overnight.

Luminex analysis was conducted on splenocytes 7 days after dose 3. 1 x10⁶ cells/well were activated with 2 pg/ml of each p53 peptide (Table 16). Cells were stimulated overnight. Supernatant was collected, and tested with mouse cytokine/chemokine magnetic bead kit (See Error! Reference source not found.17) for the simultaneous quantification of the following analytes: GM-CSF, IL2, TNFa, IN Fy, and Granzyme B, as shown in FIGS. 8A-8E. Table 17. Luminex Kit

Example 5. Effects of p53 R248W peptides on in vivo killing

This experiment aimed to determine if the AMP-p53 R248W vaccine induced T cell response was capable of in vivo killing of antigen-pulsed target cells.

3 groups of 16 C57BL/6J mice each were administered a vaccine including the components of Table 19. Groups were dosed as described in Table 18. 8 mice were used for a post-dose 3 in vivo killing assay, while the remaining 8 were used for a second attempt at a post dose 5 in vivo killing assay.

Table 18. Summary of Vaccine Administration in Mice

The optimal amount of peptide-antigen was determined by previous experiments to be 1 .25 nmol of peptide per injection. Because of low solubility of some AMP-peptides, all AMP-peptide stocks were cosolubilized with AMP-CpG. To achieve this, AMP-peptides were dissolved in 50% t-butanol first, followed by the addition of 1 :4 molar ratio of AMP-CpG and subsequent lyophilization. The obtained powder was then resuspended in 1 X PBS to a concentration of close to 1 mg/ml. The vaccine components are described in Table 19. AMP-vaccine stocks were further diluted to final concentrations using 1 X PBS. AMP-vaccines contained 1 .25 nmol AMP-antigen and 5 nmol AMP-adjuvant.

The immunizations were administered subcutaneously (SC) into the tail base of female B6 mice, bilaterally with 50 pL per side. SC injections ensured that the vaccine was optimally delivered into lymph nodes via natural lymph drainage, and bi-weekly injections were determined to be optimal in immune response. Table 19. Vaccine Components

For the in-vivo killing assay, 25 B6 untreated mice were sacrificed for naive splenocytes. Those cells were split into un-pulsed and p53-peptide pulsed cells (peptides are listed in Table 20). Cells were then loaded with 0.5 pM (un-pulsed) and 5 pM (p53-pulsed) Tag-it cell trace dye and a mixture of 10 x10⁶ cells per specificity was injected into the tail vein of the vaccinated mice either PD3 (FIG. 10A) or PD5 (FIG. 10B). 24 hours after target cell transfer splenocytes were retrieved from these animals and analyzed by flow cytometry for the presence of dye-loaded target cells. Table 20. Pulsing Peptides

Example 6. Effects of amphiphilic p53 R248W peptide in comparison to soluble R248W peptide

This experiment aimed to compare the R248W peptide candidates, amphiphilic or soluble, against each other at the optimized doses. 7 groups of 10 C57BL/6J mice each were administered a vaccine including the components of

Table 22. Groups were dosed as described in Table 21 . 5 animals being analyzed post-dose 2 (PD2), and 5 animals being analyzed post-dose 3 (PD3). Table 21. Summary of Vaccine Administration in Mice

The optimal amount of peptide-antigen was determined by previous experiments to be 1 .25 nmol of peptide per injection. Because of low solubility of some AMP-peptides, all AMP-peptide stocks were cosolubilized with AMP-CpG. To achieve this, AMP-peptides were dissolved in 50% t-butanol first, followed by the addition of 1 :4 molar ratio of AMP-CpG and subsequent lyophilization. The obtained powder was then resuspended in 1 X PBS to a concentration of close to 1 mg/ml. The vaccine components are described in Table 22. AMP-vaccine stocks were further diluted to final concentrations using 1 X PBS. AMP-vaccines contained 1 .25 nmol AMP-antigen and 5 nmol AMP-adjuvant.

The immunizations were administered subcutaneously (SC) into the tail base of female B6 mice, bilaterally with 50 pL per side. SC injections ensured that the vaccine was optimally delivered into lymph nodes via natural lymph drainage, and bi-weekly injections were determined to be optimal in immune response.

Table 22. Vaccine Components

An ICS assay to assess TNFa and IFNy levels was performed on splenocytes 7 days after dose

2, and on splenocytes, lung-resident lymphocytes and PBMCs (10^A6 cells/well) 7 days after dose 3, as shown in FIG. 14A, FIG. 14B, and FIGS. 15A-15F. Cells were also stained for CD4, CD8, and CD3 using the antibodies described in Table 23. ICS samples were activated overnight (in the presence of Brefeldin A and Monensin) with 2 pg/ml of each peptide listed in Table 24.

Table 23. Antibodies used for ICS

Table 24. Re-Stimulation Peptides

ELISpot analysis for IFNy was performed on splenocytes 7 days post dose 2 as well as post dose 3, and on lung-resident lymphocytes 7 days post dose 3, as shown in FIGS. 11 A-11 D. 0.1 x10⁶ cells/well were activated either with peptide pools (listed in Table 24) or their WT counterparts of 2 pg/ml of each peptide. IFNy plates were stimulated overnight.

FluoroSpot analysis for IFNy/TNFa/IL2 was also performed on splenocytes and lung-resident lymphocytes 7 days post dose 3, as shown in FIGS. 12A and 12B. 0.1 x10⁶ splenocytes/well were activated with peptide pools (listed in Table 24) of 2 pg/ml of each peptide. The plates were stimulated overnight.

ELISpot analysis for Granzyme B was performed on splenocytes (0.1x10^A6 cells/well) 7 days post dose 3 as shown in FIG. 13. 0.1x10⁶ splenocytes/well were activated with peptide pools (listed in Table 24) of 2 pg/ml of each peptide. Plates were stimulated overnight.

Luminex analysis was conducted on splenocytes 7 days after dose 3. For PD3, 1 x10⁶ cells/well were activated with 2 pg/ml of each p53 peptide (Table 24). Cells were stimulated overnight. Supernatant was collected, and tested with mouse cytokine/chemokine magnetic bead kit (see, Error! Reference source not found. 25) for the simultaneous quantification of the following analytes: GM-CSF, IL2, TNFa, INFy, and Granzyme B, as shown in FIGS. 16A-16E. Table 25. Luminex Kit

Example 7. Effect of amphiphilic p53 R248W peptide in transgenic mice

This experiment aimed to determine the effect of the amphiphilic p53 R248W peptide vaccine in HLA transgenic mice (transgenic for HLA-A2 and HLA-DR1 and deleted for both H-2 class 1 and 2 molecules (p2m ^z- H-2Db ^z- 1 Ap ^z IAcr lEp ).

3 groups of HLA-transgenic mice with 5 mice in the Mock group, 5 mice vaccinated with soluble peptide, and 10 mice vaccinated with the amphiphilic peptide; each mouse was administered a vaccine including the components of Table 27. Groups were dosed and analyzed post dose 5 as described in Table 26.

Table 26. Summary of Vaccine Administration in Mice

The optimal amount of peptide-antigen was determined by previous experiments to be 1 .25 nmol of peptide per injection. Because of low solubility of some AMP-peptides, all AMP-peptide stocks were cosolubilized with AMP-CpG. To achieve this, AMP-peptides were dissolved in 50% t-butanol first, followed by the addition of 1 :4 molar ratio of AMP-CpG and subsequent lyophilization. The obtained powder was then resuspended in 1 X PBS to a concentration of close to 1 mg/ml. The vaccine components are described in Table 27. AMP-vaccine stocks were further diluted to final concentrations using 1 X PBS. AMP-vaccines contained 1 .25 nmol AMP-antigen and 5 nmol AMP-adjuvant.

The immunizations were administered subcutaneously (SC) into the tail base of HLA-transgenic mice, bilaterally with 50 pL per side. SC injections ensured that the vaccine was optimally delivered into lymph nodes via natural lymph drainage, and bi-weekly injections were determined to be optimal in immune response. Table 27. Vaccine Components

ELISpot analysis for IFNy was performed on splenocytes 7 days post dose 5 with 0.2x10⁶ cells/well and 2 pg/ml of each peptide, as shown in FIG. 17. Splenocytes were activated with the p53 peptides described in Table 28. IFNy plates were stimulated overnight.

Table 28. Stimulating Peptides

Luminex analysis was conducted on splenocytes 7 days after dose 5. 1 x10⁶ cells/well were activated with 2 pg/ml of each p53 peptide (Table 28). Cells were stimulated overnight. Supernatant was collected, and tested with mouse cytokine/chemokine magnetic bead kit (See Error! Reference source not found. 29) for the simultaneous quantification of the following analytes: GM-CSF, IL2, TNFa, I NFy, and Granzyme B, as shown in FIGS. 18A-18E.

Table 29. Luminex kit

Half of the group administered amphiphilic p53 R248W peptide showed strong responses, with high expression for IFNy (ELISpot), after 5 doses. Additionally, Luminex results support the ELISpot findings, with elevated levels of IFNy, granzyme B, TNFa, and GM-CSF, and moderately higher results for IL-2 for 5 mice out of 10.

Example 8. Effects of various p53 mutant peptides to elicit an immune response

This experiment aimed to test additional p53 mutant AMP-peptides.

17 groups of 10 C57BL/6J mice each were administered a vaccine including the components of Table 31 . Groups were dosed and analyzed post dose 2 and 3 as described in Table 30. 5 animals from each group were analyzed post-dose 2 (PD2) and 5 animals from each group were analyzed post-dose 3 (PD3). Table 30. Summary of Vaccine Administration in Mice

The optimal amount of peptide-antigen was determined by previous experiments to be 1 .25 nmol of peptide per injection. Because of low solubility of some AMP-peptides, all AMP-peptide stocks were cosolubilized with AMP-CpG. To achieve this, AMP-peptides were dissolved in 50% t-butanol first, followed by the addition of 1 :4 molar ratio of AMP-CpG and subsequent lyophilization. The obtained powder was then resuspended in 1X PBS to a concentration of close to 1 mg/ml. The vaccine components are described in Table 31 . AMP-vaccine stocks were further diluted to final concentrations using 1 X PBS. AMP-vaccines contained 1 .25 nmol AMP-antigen and 5 nmol AMP-adjuvant.

Soluble peptides were prepared in 1 .1 X PBS at a concentration of 1 mg/ml and further diluted with 1 X PBS such that the final concentration of the soluble peptide was 1 .25 nmol/100 pL injection. The soluble adjuvant solutions (CpG) were prepared in limulus amebocyte lysate (LAL) H2O and further diluted with 1 X PBS such that the final concentration was 5 nmol/100 pL injection. The immunizations were administered subcutaneously (SC) into the tail base of HLA-transgenic mice, bilaterally with 50 pL per side. SC injections ensured that the vaccine was optimally delivered into lymph nodes via natural lymph drainage, and bi-weekly injections were determined to be optimal in immune response. Table 31. Vaccine Components

ELISpot analysis for IFNy was performed on splenocytes 7 days post dose 3 as shown in FIG.

19. Splenocytes (0.2x10⁶ cells/well) were activated with peptide pools listed in Table 32 in an amount of

2 pg/ml of each peptide. IFNy plates were stimulated overnight.

Table 32. Re-Stimulation Peptides

Each vaccine gets its respective peptide stim pool.

Example 9. Effect of amphiphilic p53 R248Q peptide in mice

This experiment aimed to characterize the immune response elicited upon administration of the AMP-p53 R248Q vaccine.

4 groups of 5-8 C57BL/6J mice each were administered a vaccine including the components of Table 34. Groups were dosed as described in Table 33.

Table 33. Summary of Vaccine Administration in Mice

The peptide-antigen was administered in a concentration of 1 .25 nmol or 5 nmol peptide per injection and the adjuvant was administered at a concentration of 5 nmol or 10 nmol. Because of low solubility of some AMP-peptides, all AMP-peptide stocks were co-solubilized with AMP-CpG. To achieve this, AMP-peptides were dissolved in 50% t-butanol first, followed by the addition of 1 :4 or 1 :2 molar ratio of AMP-CpG and subsequent lyophilization. The obtained powder was then resuspended in 1X PBS to a concentration of close to 1 mg/ml. The vaccine components are described in Table 34. AMP-vaccine stocks were further diluted to final concentrations using 1X PBS. AMP-vaccines contained 1 .25 nmol AMP-antigen and 5 nmol AMP-adjuvant or 5 nmol AMP-antigen and 10 nmol AMP-adjuvant.

Soluble peptides were prepared in 1 .1 X PBS at a concentration of 1 mg/ml and further diluted with 1 X PBS such that the final concentration of the soluble peptide was 1 .25 nmol/100 pL injection. The soluble adjuvant solutions (CpG) were prepared in limulus amebocyte lysate (LAL) H2O and further diluted with 1 X PBS such that the final concentration was 5 nmol/100 pL injection. The immunizations were administered subcutaneously (SC) into the tail base of female B6 mice, bilaterally with 50 pL per side. SC injections ensured that the vaccine was optimally delivered into lymph nodes via natural lymph drainage, and bi-weekly injections were determined to be optimal in immune response. Table 34. Vaccine Components

ELISpot analysis for IFNy was performed on splenocytes 7 days after dose 2 and dose 3, with 0.1 x10⁶ cells/well and 2 pg/ml of each peptide. Splenocytes were activated with the p53 peptides described in Table 36. IFNy plates were stimulated overnight. The results of this analysis are shown in FIGS. 20A-20C.

An Intracellular Stain (ICS) assay to assess TNFa and IFNy levels was performed on CD8⁺ T cells from spleen (1 x10⁶ cells/well) 7 days after dose 3, as shown in FIG. 21 C. Cells were also stained for CD4, CD8, and CD3 using the antibodies described in Table 35. ICS samples were activated overnight (in the presence of Brefeldin A and Monensin) with 2 pg/ml of each peptide listed in Table 36.

Table 35. Antibodies used for ICS

Table 36. Re-Stimulation Peptides

ELISpot analysis for Granzyme B was performed on splenocytes (0.1 x10^A6 cells/well) 7 days post dose 3 as shown in FIG. 21 A. 0.1 x10⁶ splenocytes/well were activated with peptide pools (listed in Table 36) of 2 pg/ml of each peptide. Plates were stimulated overnight.

FluoroSpot analysis for IFNy/TNFa/IL2 was also performed on splenocytes 7 days post dose 3, as shown in FIG. 21 B. 0.1 x10⁶ splenocytes/well were activated with peptide pools (listed in Table 36) of 2 pg/ml of each peptide. The plates were stimulated overnight.

Luminex analysis was conducted on splenocytes 7 days after dose 3. 1 x10⁶ cells/well were activated with 2 pg/ml of each p53 peptide (Table 36). Supernatant was collected, and tested with mouse cytokine/chemokine magnetic bead kit (See Error! Reference source not found.37) for the simultaneous quantification of the following 5 analytes: GM-CSF, IL2, TNFa, IFNy, and Granzyme B, as shown in FIGS. 22A-22E.

Table 37. Luminex kit

Example 10. Effect of amphiphilic p53 R175H peptide in mice

This experiment aimed to characterize the immune response elicited upon administration of the AMP-p53 R175H vaccine.

4 groups of 5-8 C57BL/6J mice each were administered a vaccine including the components of Table 39. Groups were dosed as described in Table 38.

Table 38. Summary of Vaccine Administration in Mice

The peptide-antigen was administered in a concentration of 1 .25 nmol or 5 nmol peptide per injection and the adjuvant was administered at a concentration of 5 nmol or 10 nmol. Because of low solubility of some AMP-peptides, all AMP-peptide stocks were co-solubilized with AMP-CpG. To achieve this, AMP-peptides were dissolved in 50% t-butanol first, followed by the addition of 1 :4 or 1 :2 molar ratio of AMP-CpG and subsequent lyophilization. The obtained powder was then resuspended in 1 X PBS to a concentration of close to 1 mg/ml. The vaccine components are described in Table 39. AMP-vaccine stocks were further diluted to final concentrations using 1 X PBS. AMP-vaccines contained 1 .25 nmol AMP-antigen and 5 nmol AMP-adjuvant or 5 nmol AMP-antigen and 10 nmol AMP-adjuvant. Soluble peptides were prepared in 1 .1 X PBS at a concentration of 1 mg/ml and further diluted with 1 X PBS such that the final concentration of the soluble peptide was 10 nmol/100 pL injection. The soluble adjuvant solutions (CpG) were prepared in limulus amebocyte lysate (LAL) H2O and further diluted with 1 X PBS such that the final concentration was 20 nmol/100 pL injection.

Table 39. Vaccine Components

ELISpot analysis for IFNy was performed on splenocytes 7 days after dose 2 and dose 3, with 0.1 x10⁶ cells/well and 2 pg/ml of each peptide. Splenocytes were activated with the p53 peptides described in Table 41 . IFNy plates were stimulated overnight. The results of this analysis are shown in FIG. 23A and FIG. 23B.

An Intracellular Stain (ICS) assay to assess TNFa and IFNy levels was performed on CD8⁺ T cells from spleen (1 x10⁶ cells/well) 7 days after dose 3, as shown in FIG. 24C. Cells were also stained for CD4, CD8, and CD3 using the antibodies described in Table 40. ICS samples were activated overnight (in the presence of Brefeldin A and Monensin) with 2 pg/ml of each peptide listed in Table 41 .

Table 40. Antibodies used for ICS

Table 41. Re-Stimulation Peptides

ELISpot analysis for Granzyme B was performed on splenocytes (0.1 x10^A6 cells/well) 7 days post dose 3 as shown in FIG. 24A. 0.1 x10⁶ splenocytes/well were activated with peptide pools (listed in Table 41 ) of 2 pg/ml of each peptide. Plates were stimulated overnight.

FluoroSpot analysis for IFNy/TNFa/IL2 was also performed on splenocytes 7 days post dose 3, as shown in FIG. 24B. 0.1 x10⁶ splenocytes/well were activated with peptide pools (listed in Table 41 ) of 2 pg/ml of each peptide. The plates were stimulated overnight.

Luminex analysis was conducted on splenocytes 7 days after dose 3. 1 x10⁶ cells/well were activated with 2 pg/ml of each p53 peptide (Table 41 ). Supernatant was collected, and tested with mouse cytokine/chemokine magnetic bead kit (See Error! Reference source not found.42) for the simultaneous quantification of the following 5 analytes: GM-CSF, IL2, TNFa, IFNy, and Granzyme B, as shown in FIGS. 25A-25E.

Table 42 Luminex kit

Example 11. Effect of amphiphilic p53 R273H peptide in mice

This experiment aimed to characterize the immune response elicited upon administration of the AMP-p53 R273H vaccine.

3 groups of 10-15 C57BL/6J mice each were administered a vaccine including the components of Table 44. Groups were dosed as described in Table 43.

Table 43. Summary of Vaccine Administration in Mice

The peptide-antigen was administered in a concentration of 1 .25 nmol peptide per injection and the adjuvant was administered at a concentration of 5 nmol. Because of low solubility of some AMP- peptides, all AMP-peptide stocks were co-solubilized with AMP-CpG. To achieve this, AMP-peptides were dissolved in 50% t-butanol first, followed by the addition of 1 :4 molar ratio of AMP-CpG and subsequent lyophilization. The obtained powder was then resuspended in 1 X PBS to a concentration of close to 1 mg/ml. The vaccine components are described in Table 44. AMP-vaccine stocks were further diluted to final concentrations using 1 X PBS. AMP-vaccines contained 1 .25 nmol AMP-antigen and 5 nmol AMP-adjuvant.

Table 44. Vaccine Components

ELISpot analysis for IFNy was performed on splenocytes 7 days after dose 2 and 3, with 0.1 x10⁶ cells/well and 2 pg/ml of each peptide. Splenocytes were activated with the p53 peptides described in Table 45. IFNy plates were stimulated overnight. The results of this analysis are shown in FIG. 26A and FIG. 26B.

Table 45. Re-Stimulation Peptides

Luminex analysis was conducted on splenocytes 7 days after dose 3. 1 x10⁶ cells/well were activated with 2 pg/ml of each p53 peptide (Table 45). Supernatant was collected, and tested with mouse cytokine/chemokine magnetic bead kit (See Error! Reference source not found.46) for the simultaneous quantification of the following 5 analytes: GM-CSF, IL2, TNFa, IFNy, and Granzyme B, as shown in FIGS. 27A-27E. Table 46 Luminex kit

For the in-vivo killing assay, 25 B6 untreated mice were sacrificed for naive splenocytes. Those cells were split into un-pulsed and p53-peptide pulsed cells (peptides are listed in Table 47). Cells were then loaded with 0.5 pM (un-pulsed) and 5 pM (p53-pulsed) Tag-it cell trace dye and a mixture of 10 x10⁶ cells per specificity was injected into the tail vein of the vaccinated mice PD4 (FIG. 28). 24 hours after target cell transfer splenocytes were retrieved from these animals and analyzed by flow cytometry for the presence of dye-loaded target cells.

Table 47. Pulsing Peptides

Example 12. Effect of amphiphilic p53 R273C peptide in mice

This experiment aimed to characterize the immune response elicited upon administration of the AMP-p53 R273C vaccine.

4 groups of 5-10 C57BL/6J mice each were administered a vaccine including the components of Table 49. Groups were dosed as described in Table 48.

Table 48. Summary of Vaccine Administration in Mice

The peptide-antigen was administered in a concentration of 1 .25 nmol or 5 nmol peptide per injection and the adjuvant was administered at a concentration of 5 nmol or 10 nmol. Because of low solubility of some AMP-peptides, all AMP-peptide stocks were co-solubilized with AMP-CpG. To achieve this, AMP-peptides were dissolved in 50% t-butanol first, followed by the addition of 1 :4 or 1 :2 molar ratio of AMP-CpG and subsequent lyophilization. The obtained powder was then resuspended in 1X PBS to a concentration of close to 1 mg/ml. The vaccine components are described in Table 49. AMP-vaccine stocks were further diluted to final concentrations using 1X PBS. AMP-vaccines contained 1 .25 nmol AMP-antigen and 5 nmol AMP-adjuvant or 5 nmol AMP-antigen and 10 nmol AMP-adjuvant.

Soluble peptides were prepared in 1 .1 X PBS at a concentration of 1 mg/ml and further diluted with 1 X PBS such that the final concentration of the soluble peptide was 10 nmol/100 pL injection. The soluble adjuvant solutions (CpG) were prepared in limulus amebocyte lysate (LAL) H2O and further diluted with 1 X PBS such that the final concentration was 20 nmol/100 pL injection.

Table 49. Vaccine Components

ELISpot analysis for IFNy was performed on splenocytes 7 days after dose 3, with 0.1 x10⁶ cells/well and 2 pg/ml of each peptide. Splenocytes were activated with the p53 peptides described in Table 50. IFNy plates were stimulated overnight. The results of this analysis are shown in FIG. 29A.

Table 50. Re-Stimulation Peptides

ELISpot analysis for Granzyme B was performed on splenocytes (0.1x10^A6 cells/well) 7 days post dose 3 as shown in FIG. 29B. 0.1 x10⁶ splenocytes/well were activated with peptide pools (listed in Table 50) of 2 pg/ml of each peptide. Plates were stimulated overnight.

FluoroSpot analysis for IFNy/TNFa/IL2 was also performed on splenocytes 7 days post dose 3, as shown in FIG. 29C. 0.1 x10⁶ splenocytes/well were activated with peptide pools (listed in Table 50) of 2 pg/ml of each peptide. The plates were stimulated overnight.

Luminex analysis was conducted on splenocytes 7 days after dose 3. 1 x10⁶ cells/well were activated with 2 pg/ml of each p53 peptide (Table 50). Supernatant was collected, and tested with mouse cytokine/chemokine magnetic bead kit (See Error! Reference source not found.51 ) for the simultaneous quantification of the following 5 analytes: GM-CSF, IL2, TNFa, IFNy, and Granzyme B, as shown in FIGS. 30A-30E. Table 51 Luminex kit

Example 13. Effect of amphiphilic p53 R282W peptide in mice

This experiment aimed to characterize the immune response elicited upon administration of the AMP-p53 R282W vaccine.

3 groups of 5-8 C57BL/6J mice each were administered a vaccine including the components of Table 53. Groups were dosed as described in Table 52.

Table 52. Summary of Vaccine Administration in Mice

The peptide-antigen was administered in a concentration of 5 nmol peptide per injection and the adjuvant was administered at a concentration of 10 nmol. Because of low solubility of some AMP- peptides, all AMP-peptide stocks were co-solubilized with AMP-CpG. To achieve this, AMP-peptides were dissolved in 50% t-butanol first, followed by the addition of 1 :2 molar ratio of AMP-CpG and subsequent lyophilization. The obtained powder was then resuspended in 1X PBS to a concentration of close to 1 mg/ml. The vaccine components are described in Table 53. AMP-vaccine stocks were further diluted to final concentrations using 1X PBS. AMP-vaccines contained 5 nmol AMP-antigen and 10 nmol AMP-adjuvant.

Table 53. Vaccine Components

ELISpot analysis for IFNy was performed on splenocytes 7 days after dose 3, with 0.1 x10⁶ cells/well and 2 pg/ml of each peptide. Splenocytes were activated with the p53 peptides described in Table 54. IFNy plates were stimulated overnight. The results of this analysis are shown in FIG. 31 A. Table 54. Re-Stimulation Peptides

ELISpot analysis for Granzyme B was performed on splenocytes (0.1x10^A6 cells/well) 7 days post dose 3 as shown in FIG. 31 B. 0.1 x10⁶ splenocytes/well were activated with peptide pools (listed in Table 54) of 2 pg/ml of each peptide. Plates were stimulated overnight.

FluoroSpot analysis for IFNy/TNFa/IL2 was also performed on splenocytes 7 days post dose 3, as shown in FIG. 31 C. 0.1 x10⁶ splenocytes/well were activated with peptide pools (listed in Table 54) of 2 pg/ml of each peptide. The plates were stimulated overnight.

Luminex analysis was conducted on splenocytes 7 days after dose 3. 1 x10⁶ cells/well were activated with 2 pg/ml of each p53 peptide (Table 54). Supernatant was collected, and tested with mouse cytokine/chemokine magnetic bead kit (See Error! Reference source not found.55) for the simultaneous quantification of the following 5 analytes: GM-CSF, IL2, TNFa, IFNy, and Granzyme B, as shown in FIGS. 32A-32E.

Table 55 Luminex kit

Example 14. Effect of amphiphilic p53 G245S peptide in mice

This experiment aimed to characterize the immune response elicited upon administration of the AMP-p53 G245S vaccine.

4 groups of 5-8 C57BL/6J mice each were administered a vaccine including the components of Table 57. Groups were dosed as described in Table 56.

Table 56. Summary of Vaccine Administration in Mice

The peptide-antigen was administered in a concentration of 1 .25 nmol or 5 nmol peptide per injection and the adjuvant was administered at a concentration of 5 nmol. Because of low solubility of some AMP-peptides, all AMP-peptide stocks were co-solubilized with AMP-CpG. To achieve this, AMP- peptides were dissolved in 50% t-butanol first, followed by the addition of 1 :4 or 1 :1 molar ratio of AMP- CpG and subsequent lyophilization. The obtained powder was then resuspended in 1X PBS to a concentration of close to 1 mg/ml. The vaccine components are described in Table 57. AMP-vaccine stocks were further diluted to final concentrations using 1X PBS. AMP-vaccines contained 1 .25 nmol AMP-antigen and 5 nmol AMP-adjuvant or 5 nmol AMP-antigen and 5 nmol AMP-adjuvant.

Soluble peptides were prepared in 1 .1 X PBS at a concentration of 1 mg/ml and further diluted with 1 X PBS such that the final concentration of the soluble peptide was 5 nmol/100 pL injection. The soluble adjuvant solutions (CpG) were prepared in limulus amebocyte lysate (LAL) H2O and further diluted with 1 X PBS such that the final concentration was 10 nmol/100 pL injection.

Table 57. Vaccine Components

ELISpot analysis for IFNy was performed on splenocytes 7 days after dose 2 and dose 3, with 0.1 x10⁶ cells/well and 2 pg/ml of each peptide. Splenocytes were activated with the p53 peptides described in Table 58. IFNy plates were stimulated overnight. The results of this analysis are shown in FIGS. 33A -33C.

Table 58. Re-Stimulation Peptides

ELISpot analysis for Granzyme B was performed on splenocytes (0.1x10^A6 cells/well) 7 days post dose 3 as shown in FIG. 34A. 0.1 x10⁶ splenocytes/well were activated with peptide pools (listed in Table 58) of 2 pg/ml of each peptide. Plates were stimulated overnight. FluoroSpot analysis for IFNy/TNFa/IL2 was also performed on splenocytes 7 days post dose 3, as shown in FIG. 34B. 0.1 x10⁶ splenocytes/well were activated with peptide pools (listed in Table 58) of 2 pg/ml of each peptide. The plates were stimulated overnight.

Luminex analysis was conducted on splenocytes 7 days after dose 3. 1 x10⁶ cells/well were activated with 2 pg/ml of each p53 peptide (Table 58). Supernatant was collected, and tested with mouse cytokine/chemokine magnetic bead kit (See Error! Reference source not found.59) for the simultaneous quantification of the following 5 analytes: GM-CSF, IL2, TNFa, IFNy, and Granzyme B, as shown in FIGS. 35A-35E.

Table 59 Luminex kit

Example 15. Effect of amphiphilic p53 R249S peptide in mice

This experiment aimed to characterize the immune response elicited upon administration of the AMP-p53 R249S vaccine.

3 groups of 5-8 C57BL/6J mice each were administered a vaccine including the components of Table 61 . Groups were dosed as described in Table 60.

Table 60. Summary of Vaccine Administration in Mice

The peptide-antigen was administered in a concentration of 5 nmol peptide per injection and the adjuvant was administered at a concentration of 10 nmol. Because of low solubility of some AMP- peptides, all AMP-peptide stocks were co-solubilized with AMP-CpG. To achieve this, AMP-peptides were dissolved in 50% t-butanol first, followed by the addition of 1 :2 molar ratio of AMP-CpG and subsequent lyophilization. The obtained powder was then resuspended in 1 X PBS to a concentration of close to 1 mg/ml. The vaccine components are described in Table 61 . AMP-vaccine stocks were further diluted to final concentrations using 1 X PBS. AMP-vaccines contained 5 nmol AMP-antigen and 10 nmol AMP-adjuvant.

Soluble peptides were prepared in 1 .1 X PBS at a concentration of 1 mg/ml and further diluted with 1 X PBS such that the final concentration of the soluble peptide was 5 nmol/100 pL injection. The soluble adjuvant solutions (CpG) were prepared in limulus amebocyte lysate (LAL) H2O and further diluted with 1 X PBS such that the final concentration was 10 nmol/100 pL injection. The immunizations were administered subcutaneously (SC) into the tail base of female B6 mice, bilaterally with 50 pL per side. SC injections ensured that the vaccine was optimally delivered into lymph nodes via natural lymph drainage, and bi-weekly injections were determined to be optimal in immune response.

Table 61. Vaccine Components

ELISpot analysis for IFNy was performed on splenocytes 7 days after dose 3, with 0.1 x10⁶ cells/well and 2 pg/ml of each peptide. Splenocytes were activated with the p53 peptides described in Table 62. IFNy plates were stimulated overnight. The results of this analysis are shown in FIG. 36A.

ELISpot analysis for Granzyme B was performed on splenocytes (0.1x10^A6 cells/well) 7 days post dose 3 as shown in FIG. 36B. 0.1 x10⁶ splenocytes/well were activated with peptide pools (listed in Table 62) of 2 pg/ml of each peptide. Plates were stimulated overnight.

FluoroSpot analysis for IFNy/TNFa/IL2 was also performed on splenocytes 7 days post dose 3, as shown in FIG. 36C. 0.1 x10⁶ splenocytes/well were activated with peptide pools (listed in Table 62) of 2 pg/ml of each peptide. The plates were stimulated overnight.

Table 62. Re-Stimulation Peptides

Example 16. Effect of amphiphilic p53 Y220C peptide in mice

This experiment aimed to characterize the immune response elicited upon administration of the AMP-p53 Y220C vaccine.

3 groups of 10-15 C57BL/6J mice each were administered a vaccine including the components of Table 64. Groups were dosed as described in Table 63. Table 63. Summary of Vaccine Administration in Mice

The peptide-antigen was administered in a concentration of 1 .25 nmol peptide per injection and the adjuvant was administered at a concentration of 5 nmol. Because of low solubility of some AMP- peptides, all AMP-peptide stocks were co-solubilized with AMP-CpG. To achieve this, AMP-peptides were dissolved in 50% t-butanol first, followed by the addition of 1 :4 molar ratio of AMP-CpG and subsequent lyophilization. The obtained powder was then resuspended in 1X PBS to a concentration of close to 1 mg/ml. The vaccine components are described in Table 64. AMP-vaccine stocks were further diluted to final concentrations using 1X PBS. AMP-vaccines contained 1 .25 nmol AMP-antigen and 5 nmol AMP-adjuvant.

Table 64. Vaccine Components

ELISpot analysis for IFNy was performed on splenocytes 7 days after dose 2 and 3, with 0.1 x10⁶ cells/well and 2 pg/ml of each peptide. Splenocytes were activated with the p53 peptides described in Table 66. IFNy plates were stimulated overnight. The results of this analysis are shown in FIG. 37A and FIG. 37B.

An Intracellular Stain (ICS) assay to assess TNFa and IFNy levels was performed on CD4⁺ T cells from spleen (1 x10⁶ cells/well) 7 days after dose 3, as shown in FIG. 37C. Cells were also stained for CD4, CD8, and CD3 using the antibodies described in Table 65. ICS samples were activated overnight (in the presence of Brefeldin A and Monensin) with 2 pg/ml of each peptide listed in Table 66.

Table 65. Antibodies used for ICS

Table 66. Re-Stimulation Peptides

Luminex analysis was conducted on splenocytes 7 days after dose 3. 1 x10⁶ cells/well were activated with 2 pg/ml of each p53 peptide (Table 66). Supernatant was collected, and tested with mouse cytokine/chemokine magnetic bead kit (See Error! Reference source not found.67) for the simultaneous quantification of the following 5 analytes: GM-CSF, IL2, TNFa, IFNy, and Granzyme B, as shown in FIGS. 38A-38E.

Table 67. Luminex kit

Example 17. Effect of amphiphilic p53 C135Y peptide in mice

This experiment aimed to characterize the immune response elicited upon administration of the AMP-p53 C135Y vaccine.

3 groups of 10 C57BL/6J mice each were administered a vaccine including the components of Table 69. Groups were dosed as described in Table 68. Table 68. Summary of Vaccine Administration in Mice

The peptide-antigen was administered in a concentration of 1 .25 nmol peptide per injection and the adjuvant was administered at a concentration of 5 nmol. Because of low solubility of some AMP- peptides, all AMP-peptide stocks were co-solubilized with AMP-CpG. To achieve this, AMP-peptides were dissolved in 50% t-butanol first, followed by the addition of 1 :4 molar ratio of AMP-CpG and subsequent lyophilization. The obtained powder was then resuspended in 1X PBS to a concentration of close to 1 mg/ml. The vaccine components are described in Table 69. AMP-vaccine stocks were further diluted to final concentrations using 1X PBS. AMP-vaccines contained 1 .25 nmol AMP-antigen and 5 nmol AMP-adjuvant.

Table 69. Vaccine Components

ELISpot analysis for IFNy was performed on splenocytes 7 days after dose 2 and 3, with 0.1 x10⁶ cells/well and 2 pg/ml of each peptide. Splenocytes were activated with the p53 peptides described in Table 71 . IFNy plates were stimulated overnight. The results of this analysis are shown in FIG. 39A and FIG. 39B.

An Intracellular Stain (ICS) assay to assess TNFa and IFNy levels was performed on CD4⁺ T cells from spleen (1 x10⁶ cells/well) 7 days after dose 3, as shown in FIG. 39C. Cells were also stained for CD4, CD8, and CD3 using the antibodies described in Table 70. ICS samples were activated overnight (in the presence of Brefeldin A and Monensin) with 2 pg/ml of each peptide listed in Table 71 .

Table 70. Antibodies used for ICS

Table 71. Re-Stimulation Peptides

Luminex analysis was conducted on splenocytes 7 days after dose 3. 1 x10⁶ cells/well were activated with 2 pg/ml of each p53 peptide (Table 71 ). Supernatant was collected, and tested with mouse cytokine/chemokine magnetic bead kit (See Error! Reference source not found.72) for the simultaneous quantification of the following 5 analytes: GM-CSF, IL2, TNFa, IFNy, and Granzyme B, as shown in FIGS. 40A-40E.

Table 72 Luminex kit

For the in-vivo killing assay, 25 B6 untreated mice were sacrificed for naive splenocytes. Those cells were split into un-pulsed and p53-peptide pulsed cells (peptides are listed in Table 73). Cells were then loaded with 0.5 pM (un-pulsed) and 5 pM (p53-pulsed) Tag-it cell trace dye and a mixture of 10 x10⁶ cells per specificity was injected into the tail vein of the vaccinated mice PD4 (FIG. 41 ). 24 hours after target cell transfer splenocytes were retrieved from these animals and analyzed by flow cytometry for the presence of dye-loaded target cells. Table 73. Pulsing Peptides

Example 18. Effect of amphiphilic p53 R158H peptide in mice

This experiment aimed to characterize the immune response elicited upon administration of the AMP-p53 R158H vaccine.

3 groups of 10 C57BL/6J mice each were administered a vaccine including the components of Table 75. Groups were dosed as described in Table 74.

Table 74. Summary of Vaccine Administration in Mice

The peptide-antigen was administered in a concentration of 1 .25 nmol peptide per injection and the adjuvant was administered at a concentration of 5 nmol. Because of low solubility of some AMP- peptides, all AMP-peptide stocks were co-solubilized with AMP-CpG. To achieve this, AMP-peptides were dissolved in 50% t-butanol first, followed by the addition of 1 :4 molar ratio of AMP-CpG and subsequent lyophilization. The obtained powder was then resuspended in 1X PBS to a concentration of close to 1 mg/ml. The vaccine components are described in Table 75. AMP-vaccine stocks were further diluted to final concentrations using 1X PBS. AMP-vaccines contained 1 .25 nmol AMP-antigen and 5 nmol AMP-adjuvant.

Table 75. Vaccine Components

ELISpot analysis for IFNy was performed on splenocytes 7 days after dose 2 and dose 3, with 0.1 x10⁶ cells/well and 2 pg/ml of each peptide. Splenocytes were activated with the p53 peptides described in Table 76. IFNy plates were stimulated overnight. The results of this analysis are shown in FIG. 42A and FIG. 42B.

Table 76. Re-Stimulation Peptides

Example 19. Effect of amphiphilic p53 H214R peptide in mice

This experiment aimed to characterize the immune response elicited upon administration of the AMP-p53 H214R vaccine.

3 groups of 10-15 C57BL/6J mice each were administered a vaccine including the components of Table 78. Groups were dosed as described in Table 77.

Table 77. Summary of Vaccine Administration in Mice

The peptide-antigen was administered in a concentration of 1 .25 nmol peptide per injection and the adjuvant was administered at a concentration of 5 nmol. Because of low solubility of some AMP- peptides, all AMP-peptide stocks were co-solubilized with AMP-CpG. To achieve this, AMP-peptides were dissolved in 50% t-butanol first, followed by the addition of 1 :4 molar ratio of AMP-CpG and subsequent lyophilization. The obtained powder was then resuspended in 1X PBS to a concentration of close to 1 mg/ml. The vaccine components are described in Table 78. AMP-vaccine stocks were further diluted to final concentrations using 1X PBS. AMP-vaccines contained 1 .25 nmol AMP-antigen and 5 nmol AMP-adjuvant.

Table 78. Vaccine Components

ELISpot analysis for IFNy was performed on splenocytes 7 days after dose 2 and dose 3, with 0.1 x10⁶ cells/well and 2 pg/ml of each peptide. Splenocytes were activated with the p53 peptides described in Table 79. IFNy plates were stimulated overnight. The results of this analysis are shown in FIG. 43A and FIG. 43B.

Table 79. Re-Stimulation Peptides

ELISpot analysis for Granzyme B was performed on splenocytes (0.1x10^A6 cells/well) 7 days post dose 3 as shown in FIG. 44A. 0.1 x10⁶ splenocytes/well were activated with peptide pools (listed in Table 79) of 2 pg/ml of each peptide. Plates were stimulated overnight.

FluoroSpot analysis for IFNy/TNFa/IL2 was also performed on splenocytes 7 days post dose 3, as shown in FIG. 44B. 0.1 x10⁶ splenocytes/well were activated with peptide pools (listed in Table 79) of 2 pg/ml of each peptide. The plates were stimulated overnight.

Luminex analysis was conducted on splenocytes 7 days after dose 3. 1 x10⁶ cells/well were activated with 2 pg/ml of each p53 peptide (Table 79). Supernatant was collected, and tested with mouse cytokine/chemokine magnetic bead kit (See Error! Reference source not found.80) for the simultaneous quantification of the following 5 analytes: GM-CSF, IL2, TNFa, IFNy, and Granzyme B, as shown in FIGS. 45A-45E.

Table 80 Luminex kit

Example 20. Effect of amphiphilic p53 wildtype peptide in mice

This experiment aimed to characterize the immune response elicited upon administration of the AMP-p53 wildtype vaccine.

3 groups of 10 C57BL/6J mice each were administered a vaccine including the components of Table 82. Groups were dosed as described in Table 81 .

Table 81. Summary of Vaccine Administration in Mice

The peptide-antigen was administered in a concentration of 1 .25 nmol peptide per injection and the adjuvant was administered at a concentration of 5 nmol. Because of low solubility of some AMP- peptides, all AMP-peptide stocks were co-solubilized with AMP-CpG. To achieve this, AMP-peptides were dissolved in 50% t-butanol first, followed by the addition of 1 :4 molar ratio of AMP-CpG and subsequent lyophilization. The obtained powder was then resuspended in 1X PBS to a concentration of close to 1 mg/ml. The vaccine components are described in Table 82. AMP-vaccine stocks were further diluted to final concentrations using 1X PBS. AMP-vaccines contained 1 .25 nmol AMP-antigen and 5 nmol AMP-adjuvant.

The immunizations were administered subcutaneously (SC) into the tail base of female B6 mice, bilaterally with 50 pL per side. SC injections ensured that the vaccine was optimally delivered into lymph nodes via natural lymph drainage, and bi-weekly injections were determined to be optimal in immune response. Table 82. Vaccine Components

ELISpot analysis for IFNy was performed on splenocytes 7 days after dose 2 and dose 3, with 0.1 x10⁶ cells/well and 2 pg/ml of each peptide. Splenocytes were activated with the p53 peptides described in Table 84. IFNy plates were stimulated overnight. The results of this analysis are shown in FIG. 46A and FIG. 46B.

An Intracellular Stain (ICS) assay to assess TNFa and IFNy levels was performed on CD4⁺ CD8⁺ T cells from spleen (1 x10⁶ cells/well) 7 days after dose 3, as shown in FIG. 47A and FIG. 47B, respectively. Cells were also stained for CD4, CD8, and CD3 using the antibodies described in Table 83. ICS samples were activated overnight (in the presence of Brefeldin A and Monensin) with 2 pg/ml of each peptide listed in Table 84.

Table 83. Antibodies used for ICS

Table 84. Re-Stimulation Peptides

Luminex analysis was conducted on splenocytes 7 days after dose 3. 1 x10⁶ cells/well were activated with 2 pg/ml of each p53 peptide (Table 84). Supernatant was collected, and tested with mouse cytokine/chemokine magnetic bead kit (See Error! Reference source not found.85) for the simultaneous quantification of the following 5 analytes: GM-CSF, IL2, TNFa, IFNy, and Granzyme B, as shown in FIGS. 48A-48E. Table 85 Luminex kit

Other Embodiments Various modifications and variations of the described compositions, methods, and uses of the invention will be apparent to those skilled in the art without departing from the scope and spirit of the invention. Although the invention has been described in connection with specific embodiments, it should be understood that the invention as claimed should not be unduly limited to such specific embodiments. Indeed, various modifications of the described modes for carrying out the invention that are obvious to those skilled in the art are intended to be within the scope of the invention.

All publications, patents, and patent applications are herein incorporated by reference in their entirety to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference in its entirety.

What is claimed is:

Claims

1 . A compound comprising an albumin-binding domain and a mutant p53 peptide, or a pharmaceutically acceptable salt thereof.

2. The compound or pharmaceutically acceptable salt thereof of claim 1 , wherein the mutant p53 peptide is a 5 to 50 amino acid fragment of SEQ ID NO: 1 comprising at least one amino acid substitution relative to the corresponding sequence of SEQ ID NO:1 .

3. The compound or pharmaceutically acceptable salt thereof of claim 1 , wherein the mutant p53 peptide is a 20 to 40 amino acid fragment of SEQ ID NO: 1 comprising at least one amino acid substitution relative to the corresponding sequence of SEQ ID NO:1 .

4. The compound or pharmaceutically acceptable salt thereof of claim 3, wherein the mutant p53 peptide is a 20 to 30 amino acid fragment of SEQ ID NO: 1 comprising at least one amino acid substitution relative to the corresponding sequence of SEQ ID NO: 1 .

5. The compound or pharmaceutically acceptable salt thereof of claim 4, wherein the mutant p53 peptide is a 30 amino acid fragment of SEQ ID NO: 1 comprising at least one amino acid substitution relative to the corresponding sequence of SEQ ID NO: 1 .

6. The compound or pharmaceutically acceptable salt thereof of any one of claims 1 -5, wherein the mutant p53 peptide comprises at least one of the amino acid substitutions described in TABLE 1 .

7. The compound or pharmaceutically acceptable salt thereof of claim 1 , wherein the mutant p53 peptide consists of the amino acid sequence of YMCNSSCMGGMNWRPILTIITLEDS (SEQ ID NO: 2).

8. The compound or pharmaceutically acceptable salt thereof of claim 1 , wherein the mutant p53 peptide consists of the amino acid sequence of YMCNSSCMGGMNQRPILTIITLEDS (SEQ ID NO:3).

9. The compound or pharmaceutically acceptable salt thereof of claim 1 , wherein the mutant p53 peptide comprises a 10 amino acid fragment of SEQ ID NO: 2 or 3.

10. The compound or pharmaceutically acceptable salt thereof of claim 9, wherein the mutant p53 peptide comprises the amino acid sequence NWRPILTIIT (SEQ ID NO: 46) or NQRPILTIIT (SEQ ID NO: 47).

11 . The compound or pharmaceutically acceptable salt thereof of claim 1 , wherein the mutant p53 peptide comprises a 10, 18, or 25 amino acid fragment of SEQ ID NO: 2 or 3.

12. The compound or pharmaceutically acceptable salt thereof of claim 11 , wherein the mutant p53 peptide comprises the amino acid sequence NSSCMGGMNWRPILTIIT (SEQ ID NO: 48) or NSSCMGGMNQRPILTIIT (SEQ ID NO: 49).

13. The compound or pharmaceutically acceptable salt thereof of claim 1 , wherein the mutant p53 peptide consists of the amino acid sequence of DSSGNLLGRNSFEVHVCACPGRDRRTEEEN (SEQ ID NO: 99), or a fragment thereof.

14. The compound or pharmaceutically acceptable salt thereof of claim 1 , wherein the mutant p53 peptide consists of the amino acid sequence of DSSGNLLGRNSFEVCVCACPGRDRRTEEEN (SEQ ID NO: 113), or a fragment thereof.

15. The compound or pharmaceutically acceptable salt thereof of claim 1 , wherein the mutant p53 peptide consists of the amino acid sequence of NSFEVCVCACPGRDWRTEEENLRKKGEPHH (SEQ ID NO: 114), or a fragment thereof.

16. The compound or pharmaceutically acceptable salt thereof of claim 1 , wherein the mutant p53 peptide consists of the amino acid sequence of TIHYNYMCNSSCMGSMNRRPILTIITLEDS (SEQ ID NO: 115), or a fragment thereof.

17. The compound or pharmaceutically acceptable salt thereof of claim 1 , wherein the mutant p53 peptide consists of the amino acid sequence of NYMCNSSCMGGMNRSPILTIITLEDSSGNL (SEQ ID NO: 116), or a fragment thereof.

18. The compound or pharmaceutically acceptable salt thereof of claim 1 , wherein the mutant p53 peptide consists of the amino acid sequence of LDDRNTFRHSVVVPCEPPEVGSDCTTIHYN (SEQ ID NO: 117), or a fragment thereof.

19. The compound or pharmaceutically acceptable salt thereof of claim 1 , wherein the mutant p53 peptide consists of the amino acid sequence of RLGFLHSGTAKSVTCTYSPALNKMFYQLAK (SEQ ID NO: 118), or a fragment thereof.

20. The compound or pharmaceutically acceptable salt thereof of claim 1 , wherein the mutant p53 peptide consists of the amino acid sequence of PVQLWVDSTPPPGTRVHAMAIYKQSQHMTE (SEQ ID NO: 119), or a fragment thereof.

21 . The compound or pharmaceutically acceptable salt thereof of claim 1 , wherein the mutant p53 peptide consists of the amino acid sequence of LRVEYLDDRNTFRRSVVVPYEPPEVGSDCT (SEQ ID NO: 120), or a fragment thereof.

22. The compound or pharmaceutically acceptable salt thereof of claim 1 , wherein the mutant p53 peptide consists of the amino acid sequence of RAMAIYKQSQHMTEVVRHCPHHERCSDSDG (SEQ ID NO: 122), or a fragment thereof.

23. A compound comprising an albumin-binding domain and a p53 peptide, or a pharmaceutically acceptable salt thereof, wherein the p53 peptide comprises the sequence MAIYKQSQHMTEVVRRCPHHERCSDSDGLAP (SEQ ID NO:100), or a fragment thereof.

24. The compound or pharmaceutically acceptable salt thereof of claim 23, wherein the fragment comprises the sequence AIYKQSQHM (SEQ ID NO:101 ).

25. A compound comprising an albumin-binding domain and a p53 peptide, or a pharmaceutically acceptable salt thereof, wherein the p53 peptide comprises the sequence EGNLRVEYLDDRNTFRHSVVVPCEPPEVGSD (SEQ ID NQ:102), or a fragment thereof.

26. The compound or pharmaceutically acceptable salt thereof of claim 25, wherein the fragment comprises the sequence EYLDDRNTF (SEQ ID NO:103).

27. A compound comprising an albumin-binding domain and a p53 peptide, or a pharmaceutically acceptable salt thereof, wherein the p53 peptide comprises the sequence YLDDRNTFRHSVVVPYEPPEVGSDCTTIHYN (SEQ ID NO:104) or a fragment thereof.

28. The compound or pharmaceutically acceptable salt thereof of claim 27, wherein the fragment comprises the sequence VVPYEPPEV (SEQ ID NO:105).

29. A compound comprising an albumin-binding domain and a p53 peptide, or a pharmaceutically acceptable salt thereof, wherein the p53 peptide comprises the sequence TTIHYNYMCNSSCMGGMNRRPILTIITLEDSSGNL (SEQ ID NQ:106) or a fragment thereof.

30. A compound comprising an albumin-binding domain and a p53 peptide, or a pharmaceutically acceptable salt thereof, wherein the p53 peptide comprises the sequence EDSSGNLLGRNSFEVRVCACPGRDRRTEEENLRKK (SEQ ID NO:107), or a fragment thereof.

31 . The compound or pharmaceutically acceptable salt thereof of claim 30, wherein the fragment comprises the sequence LLGRNSFEV (SEQ ID NQ:108).

32. A compound comprising an albumin-binding domain and a p53 peptide, or a pharmaceutically acceptable salt thereof, wherein the p53 peptide comprises the sequence RLGFLHSGTAKSVTC (SEQ ID NO:109), or a fragment thereof.

33. A compound comprising an albumin-binding domain and a p53 peptide, or a pharmaceutically acceptable salt thereof, wherein the p53 peptide comprises the sequence STPPPGTRV (SEQ ID NO:110), or a fragment thereof.

34. A compound comprising an albumin-binding domain and a p53 peptide, or a pharmaceutically acceptable salt thereof, wherein the p53 peptide comprises the sequence TYPALNKMF (SEQ ID NO:111 ), or a fragment thereof.

35. A compound comprising an albumin-binding domain and a p53 peptide, or a pharmaceutically acceptable salt thereof, wherein the p53 peptide comprises the sequence RMPEAAPPV (SEQ ID NO:112), or a fragment thereof.

36. A compound comprising an albumin-binding domain and a p53 peptide, or a pharmaceutically acceptable salt thereof, wherein the p53 peptide comprises the sequence TEDPGPDEAPRMPEAAPPVAPAPAAPTPAA (SEQ ID NO:121 ), or a fragment thereof.

37. The compound or pharmaceutically acceptable salt thereof of any one of claims 1 -36, wherein the peptide comprises an N-terminal modification.

38. The compound or pharmaceutically acceptable salt thereof of claim 37, wherein the N-terminal modification is the addition of an acetylcysteine.

39. The compound or pharmaceutically acceptable salt thereof of claim 37, wherein the N-terminal modification is the addition of a des-aminocysteine homolog.

40. The compound or pharmaceutically acceptable salt thereof of claim 39, wherein the des- aminocysteine homolog is 3-mercaptopropionic acid or mercaptoacetic acid.

41 . The compound or pharmaceutically acceptable salt thereof of any one of claims 1 -40, wherein the albumin-binding comprises a lipid.

42. The compound or pharmaceutically acceptable salt thereof of claim 41 , wherein the lipid is a diacyl lipid.

43. The compound or pharmaceutically acceptable salt thereof of claim 42, wherein the diacyl lipid comprises acyl chains comprising 12-30 hydrocarbon units, 14-25 hydrocarbon units, 16-20 hydrocarbon units, or 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, or 30 hydrocarbon units.

44. The compound or pharmaceutically acceptable salt thereof of claim 42 or 43, wherein the lipid is 1 ,2- distearoyl-sn-glycero-3-phosphoethanolamine (DSPE).

45. The compound or pharmaceutically acceptable salt thereof of any one of claims 1 -44, wherein the peptide is bonded or linked by a linker to the following lipid:

or a salt thereof, wherein X is O or S.

46. The compound or pharmaceutically acceptable salt thereof of claim 45, wherein the linker is selected from the group consisting of a hydrophilic polymer, a string of hydrophilic amino acids, a polysaccharide, and an oligonucleotide, or a combination thereof.

47. The compound or pharmaceutically acceptable salt thereof of claim 46, wherein the linker comprises "N" polyethylene glycol units, wherein N is between 24-50.

48. The compound or pharmaceutically acceptable salt thereof of claim 47, wherein the linker comprises PEG24-amido-PEG24.

49. A method of inducing an immune response in a subject, the method comprising administering a compound or pharmaceutically acceptable salt thereof of any one of claims 1 -48 to the subject.

50. The method of claim 49, further comprising administering an adjuvant to the subject.

51 . The method of claim 49 or 50, wherein the compound or pharmaceutically acceptable salt thereof is administered subcutaneously, intramuscularly, intravenously, or transmucosally.

52. The method of any one of claims 49-51 , wherein the subject is a mammal.

53. The method of claim 52, wherein the subject is a human.

54. A compound or pharmaceutically acceptable salt thereof comprising an albumin-binding domain and a p53 peptide or mutant p53 peptide for use in a method of inducing an immune response in a subject, wherein the method comprises administering the compound comprising the compound of any one of claims 1 -48 to the subject.

55. The compound or pharmaceutically acceptable salt thereof for use according to claim 54, further comprising administering an adjuvant to the subject.

56. The compound or pharmaceutically acceptable salt thereof for use according to claim 54 or 55, wherein the compound or pharmaceutically acceptable salt thereof is formulated for subcutaneous, intramuscular, intravenous, or transmucosal administration.

57. The compound or pharmaceutically acceptable salt thereof for use according to any one of claims 54- 56 wherein the subject is a mammal.

58. The compound or pharmaceutically acceptable salt thereof for use according to claim 57, wherein the subject is a human.

59. A pharmaceutical composition comprising compound or pharmaceutically acceptable salt thereof of any one of claims 1 -48 and a pharmaceutically acceptable carrier.

60. A kit comprising a compound or pharmaceutically acceptable salt thereof of any one of claims 1 -48 or the pharmaceutical composition of claim 59 and instructions for administration.