Self-assembled nanoparticles

Abstract

The present disclosure relates to a vaccine comprising at least one peptide antigen conjugate having the formula selected from the group consisting of PEG-[E1]-A-[E2]-[U]-H and H-[U]-[E1]-A-[E2]-PEG wherein E1 is the N-terminal extension, E2 is the C-terminal extension, A is a peptide antigen, H is a hydrophobic block wherein one or more drug molecules (D) are optionally attached to each H either directly or via a suitable linker X1; u is a linker, [] represents a group being optional, and-represents that two adjacent groups are attached to each other directly via a covalent bond or indirectly via a suitable linker X. The vaccine can be used to treat or prevent cancer, autoimmune disease, allergy or infectious disease.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims priority to U.S. Provisional Application No. 63 / 380,931, filed on October 25, 2022, the disclosure of which is hereby incorporated by reference in its entirety.

[0003] This invention was made under a cooperative research and development agreement with the National Institutes of Health, U.S. Department of Health and Human Services. The U.S. Government has certain rights in this invention. Technical Field

[0004] The present disclosure relates to novel PEGylated peptide antigen conjugate compositions that can be used to form nanoparticles, including micellar structures or polymersomes; methods for making the PEGylated peptide antigen conjugate compositions; processes for formulating drug molecules with the PEGylated peptide antigen conjugate compositions that form nanoparticles; and therapeutic uses of the nanoparticles for drug delivery.

[0005] Reference to a sequence listing

[0006] Pursuant to 35 USC § 1.52(e), a sequence listing submitted in XML format is filed with this application and is hereby incorporated by reference. The name of the ASCII text file of the sequence listing is VNA002WO.xml, the creation date of the file is October 24, 2023, and the size of the file is 199 KB. Background Art

[0007] Various peptide-based vaccine technologies are known and have been developed for delivering peptide antigens to induce an immune response. U.S. Patent Application No. 2020 / 0054741 relates to novel peptide-based vaccines, methods for making such novel peptide-based vaccines, and uses of such vaccines for delivering peptide antigens to induce an immune response. U.S. Patent Application No. 2021 / 0113705 discloses improved methods for making peptide-based vaccines.

[0008] Pegylation has been widely used as a means of shielding molecules from immune system attacks. The FDA has approved a variety of pegylated recombinant proteins (Ramos-de-la-Peña, AM et al. International Journal of Peptide Research and Therapeutics, 2020, 26: 333–348) based on improved pharmacokinetics, which are attributed to the improved ability to escape the immune system. Pegylated liposome carriers, including DOXIL, have been developed based on similar principles of pegylation to reduce immune recognition. Our previous results on pegylated peptide antigen conjugates as disclosed in U.S. Patent Application No. 2020 / 0054741 indicate that this pegylation may be harmful to the immune response generated using peptide antigens.

[0009] Recently, there has been increasing interest in the use of peptide-based vaccines for inducing tolerance for the treatment of autoimmunity. WO 2022 / 177993 relates to a vaccine comprising a novel amphiphilic composition and at least one peptide antigen conjugate with a charged block, wherein the amphiphilic and / or at least one peptide antigen conjugate comprises a dendron amplifier. A potential challenge is that peptide antigen conjugates with charged blocks are found to cause dose-dependent hemolysis of erythrocytes. Although amphiphilic carriers have been introduced to overcome the problem, there is still a related decrease in hydrodynamic stability. Therefore, there is currently a need for improved peptide antigen conjugate compositions having reduced hemolytic activity while maintaining hydrodynamic stability, and for improved methods of delivering multiple immunomodulators and peptide antigens in particles for inducing tolerance. The goal of the present disclosure is to provide improved compositions, vaccine manufacturing methods, and their use for inducing an immune response that address the aforementioned challenges. Summary of the Invention

[0010] The present disclosure provides novel PEGylated peptide antigen conjugate compositions that can be used to form nanoparticles, including micellar structures or polymer vesicles; methods for making the PEGylated peptide antigen conjugate compositions; processes for formulating drug molecules with the PEGylated peptide antigen conjugate compositions that form nanoparticles; and therapeutic uses of the nanoparticles for drug delivery.

[0011] In a first aspect of the present disclosure, a vaccine is provided, comprising at least one peptide antigen conjugate having a formula selected from the group consisting of PEG-[E1]-A-[E2]-[U]-H and H-[U]-[E1]-A-[E2]-PEG

[0012] in

[0013] A is a peptide antigen;

[0014] E1 is the N-terminal extension;

[0015] E2 is the C-terminal extension;

[0016] H is independently a hydrophobic block at each occurrence, wherein one or more drug molecules (D) are optionally attached to each H directly or via a suitable linker X1;

[0017] U is independently a linker at each occurrence;

[0018] [ ] indicates that the group is optional, and

[0019] - means that two adjacent groups are directly attached to each other by a covalent bond or indirectly attached to each other via a suitable linker X.

[0020] In a second aspect of the present disclosure, a vaccine is provided, comprising at least one peptide antigen conjugate having a formula selected from PEG-[E1]-A-[E2]-[U]-H and H-[U]-[E1]-A-[E2]-PEG, and an amphiphile having the formula S-[B]-[U]-H,

[0021] in

[0022] A is a peptide antigen;

[0023] E1 is the N-terminal extension;

[0024] E2 is the C-terminal extension;

[0025] H is independently a hydrophobic block at each occurrence, wherein one or more drug molecules (D) are optionally attached to each H directly or via a suitable linker X1;

[0026] S is the solubilization block;

[0027] B is a spacer;

[0028] U is independently a linker at each occurrence;

[0029] [ ] indicates that the group is optional, and

[0030] - means that two adjacent groups are directly attached to each other by a covalent bond or indirectly attached to each other via a suitable linker X.

[0031] In one embodiment of the vaccine, S of the amphiphile comprises a dendrimer amplifier.

[0032] In a third aspect of the present disclosure, there is provided a vaccine for inducing tolerance, the vaccine comprising at least one peptide antigen conjugate having a formula selected from PEG-[E1]-A-[E2]-[U]-H and H-[U]-[E1]-A-[E2]-PEG, and an amphiphile having the formula S-[B]-[U]-H,

[0033] in

[0034] A is a peptide antigen;

[0035] E1 is the N-terminal extension;

[0036] E2 is the C-terminal extension;

[0037] H is independently a hydrophobic block at each occurrence, wherein one or more drug molecules (D) are optionally attached to each H directly or via a suitable linker X1;

[0038] S is the solubilization block;

[0039] B is a spacer;

[0040] U is independently a linker at each occurrence;

[0041] [ ] indicates that the group is optional,

[0042] - means that two adjacent groups are directly attached to each other by a covalent bond or indirectly attached to each other via a suitable linker X;

[0043] wherein the amphiphile comprises a dendrimer amplifier; and

[0044] The at least one peptide antigen A is selected from the group consisting of an autoantigen, an alloantigen, and an allergen.

[0045] In a fourth aspect of the present disclosure, provided herein is a vaccine comprising at least one peptide antigen conjugate having a formula selected from PEG-[E1]-A-[E2]-[U]-H and H-[U]-[E1]-A-[E2]-PEG, and an amphiphile having the formula S-[B]-[U]-H,

[0046] in

[0047] A is a peptide antigen;

[0048] E1 is the N-terminal extension;

[0049] E2 is the C-terminal extension;

[0050] H is independently a hydrophobic block at each occurrence, wherein one or more drug molecules (D) are optionally attached to each H directly or via a suitable linker X1;

[0051] S is the solubilization block;

[0052] B is a spacer;

[0053] U is independently a linker at each occurrence;

[0054] [ ] indicates that the group is optional,

[0055] - means that two adjacent groups are directly attached to each other by a covalent bond or indirectly attached to each other via a suitable linker X;

[0056] wherein the amphiphile comprises a dendrimer amplifier; and

[0057] Wherein at least one A comprises a sequence in which one or more cysteine ​​residues have been replaced by α-aminobutyric acid and / or one or more methionine residues have been replaced by norleucine.

[0058] In a fifth aspect of the present disclosure, a vaccine is provided, comprising at least one peptide antigen (A), wherein at least one A comprises a sequence in which one or more cysteine ​​residues have been replaced by α-aminobutyric acid and / or one or more methionine residues have been replaced by norleucine.

[0059] In one embodiment of the present disclosure, a vaccine is provided, wherein at least one peptide antigen (A) comprises α-aminobutyric acid and / or norleucine.

[0060] In a sixth aspect of the present disclosure, there is provided a vaccine for inducing tolerance, the vaccine comprising at least one peptide antigen conjugate having a formula selected from PEG-[E1]-A-[E2]-[U]-H and H-[U]-[E1]-A-[E2]-PEG, and an amphiphile having the formula S-[B]-[U]-H,

[0061] in

[0062] A is a peptide antigen;

[0063] E1 is the N-terminal extension;

[0064] E2 is the C-terminal extension;

[0065] H is independently a hydrophobic block at each occurrence, wherein one or more drug molecules (D) are optionally attached to each H directly or via a suitable linker X1;

[0066] S is the solubilization block;

[0067] B is a spacer;

[0068] U is independently a linker at each occurrence;

[0069] [ ] indicates that the group is optional,

[0070] - means that two adjacent groups are directly attached to each other by a covalent bond or indirectly attached to each other via a suitable linker X;

[0071] wherein the amphiphile comprises a dendrimer amplifier; and

[0072] At least one A is selected from the group consisting of an autoantigen, an alloantigen, and an allergen, and at least one D is present.

[0073] In one embodiment of the vaccine for inducing tolerance, the at least one D is selected from inhibitors of mTOR, RORγt, CDK8 / 19 and HDAC, and inhibitors of AHR, RAR and A 2a agonists.

[0074] In another embodiment of said vaccine for inducing tolerance, said at least one D is selected from ATP-competitive mTOR inhibitors.

[0075] In some embodiments of the vaccine for inducing tolerance, the at least one D is selected from AZD-8055, AZD-2016, KU-0063794, CC223, Torin-1, Torin-2, INK-128, WYE354, WYE132, OSI-027, OXA-01, PI-103, NVP-BEZ235, GNE-493, GSK2126458, rapamycin, tacrolimus, everolimus, RAD001, CCI-779 and AP23573.

[0076] In a seventh aspect of the present disclosure, there is provided a peptide antigen conjugate having a formula selected from PEG-[E1]-A-[E2]-[U]-H and H-[U]-[E1]-A-[E2]-PEG, wherein

[0077] A is a peptide antigen;

[0078] E1 is the N-terminal extension;

[0079] E2 is the C-terminal extension;

[0080] H is independently a hydrophobic block at each occurrence, wherein one or more drug molecules (D) are optionally attached to each H directly or via a suitable linker X1;

[0081] U is independently a linker at each occurrence;

[0082] [ ] indicates that the group is optional, and

[0083] - means that two adjacent groups are directly attached to each other by a covalent bond or indirectly attached to each other via a suitable linker.

[0084] In an eighth aspect of the present disclosure, provided herein is a method of treating or preventing an inflammatory disease in a subject in need thereof, the method comprising administering to the subject a vaccine comprising at least one peptide antigen conjugate having a formula selected from the group consisting of PEG-[E1]-A-[E2]-[U]-H and H-[U]-[E1]-A-[E2]-PEG, and an amphiphile having the formula S-[B]-[U]-H,

[0085] in

[0086] A is a peptide antigen;

[0087] E1 is the N-terminal extension;

[0088] E2 is the C-terminal extension;

[0089] H is independently a hydrophobic block at each occurrence, wherein one or more drug molecules (D) are optionally attached to each H directly or via a suitable linker X1;

[0090] S is the solubilization block;

[0091] B is a spacer;

[0092] U is independently a linker at each occurrence;

[0093] [ ] indicates that the group is optional,

[0094] - means that two adjacent groups are attached to each other directly by a covalent bond or indirectly via a suitable linker; and

[0095] wherein the amphiphile comprises a dendrimer amplifier and the at least one peptide antigen is selected from the group consisting of a self antigen and a foreign antigen.

[0096] In one embodiment of the method of treating an autoimmune disease, the vaccine is administered intravenously, subcutaneously, or intramuscularly.

[0097] In a ninth aspect of the present disclosure, provided herein is a method for enhancing the efficacy and / or tolerability of a vaccine, the method comprising administering to the subject a vaccine comprising at least one peptide antigen conjugate having a formula selected from the group consisting of PEG-[E1]-A-[E2]-[U]-H and H-[U]-[E1]-A-[E2]-PEG, and an amphiphile having the formula S-[B]-[U]-H,

[0098] in

[0099] E1 is the N-terminal extension;

[0100] E2 is the C-terminal extension;

[0101] H is independently a hydrophobic block at each occurrence, wherein one or more drug molecules (D) are optionally attached to each H directly or via a suitable linker X1;

[0102] S is the solubilization block;

[0103] B is a spacer;

[0104] U is independently a linker at each occurrence;

[0105] [ ] indicates that the group is optional,

[0106] - means that two adjacent groups are attached to each other directly by a covalent bond or indirectly via a suitable linker; and

[0107] wherein the amphiphile comprises a dendrimer amplifier and the at least one peptide antigen is selected from the group consisting of a self antigen and a foreign antigen.

[0108] In a tenth aspect of the present disclosure, provided herein is a method for preparing a peptide antigen conjugate having the formula PEG-[E1]-A-[E2]-[U]-H and H-[U]-[E1]-A-[E2]-PEG.

[0109] In the eleventh aspect of the present disclosure, provided herein is a method for preparing a vaccine comprising a peptide antigen conjugate of the formula PEG-[E1]-A-[E2]-[U]-H, and an amphiphile having the formula S-[B]-[U]-H,

[0110] in

[0111] A is a peptide antigen;

[0112] E1 is the N-terminal extension;

[0113] E2 is the C-terminal extension;

[0114] H is independently a hydrophobic block at each occurrence, wherein one or more drug molecules (D) are optionally attached to each H directly or via a suitable linker X1;

[0115] S is the solubilization block;

[0116] B is a spacer;

[0117] U is independently a linker at each occurrence;

[0118] [ ] indicates that the group is optional, and

[0119] - means that two adjacent groups are directly attached to each other by a covalent bond or indirectly attached to each other via a suitable linker X.

[0120] In a twelfth aspect of the present disclosure, provided herein is a vaccine for inducing tolerance, the vaccine comprising at least one peptide antigen conjugate having a formula selected from the group consisting of PEG-[E1]-A-[E2]-[U]-H-[D] and [D]-H-[U]-[E1]-A-[E2]-PEG, wherein A is a peptide antigen; E1 is an N-terminal extension; E2 is a C-terminal extension;

[0121] H is independently a hydrophobic block at each occurrence, wherein at least one drug molecule (D) is optionally attached to each H directly or via a suitable linker X1;

[0122] U is independently a linker at each occurrence;

[0123] [ ] indicates that the group is optional, and

[0124] - means that two adjacent groups are directly attached to each other by a covalent bond or indirectly attached to each other via a suitable linker X.

[0125] In some embodiments of the vaccine for inducing tolerance, when the overall average hydrophilicity of the at least one peptide antigen is >0 and / or the average peptide antigen solubility in aqueous solution at a pH between 5.5 and 8.5 is <1 mg / mL, the vaccine further comprises an amphiphile having the formula S-[B]-[U]-H, wherein S is a solubilizing block;

[0126] B is a spacer;

[0127] H is a hydrophobic block;

[0128] U is a connector;

[0129] [ ] indicates that the group is optional; and

[0130] – means that two adjacent groups are directly attached to each other by a covalent bond or indirectly attached to each other via a suitable linker X,

[0131] wherein said amphiphile S comprises a dendrimer amplifier.

[0132] In some embodiments of the vaccine for inducing tolerance, the vaccine does not contain an amphiphile having the formula S-[B]-[U]-H.

[0133] In some embodiments of the vaccine for inducing tolerance, wherein the vaccine does not contain an amphiphile having the formula S-[B]-[U]-H when the overall average hydrophilicity of the at least one peptide antigen is ≤0 and / or the average peptide antigen solubility in aqueous solution at a pH between 5.5 and 8.5 is ≥1 mg / mL.

[0134] In some embodiments of the vaccine for inducing tolerance, the vaccine comprises at least one D selected from ATP-competitive mTOR inhibitors; preferably wherein the at least one D is selected from AZD-8055, AZD-2016, KU-0063794, CC223, Torin-1, Torin-2, INK-128, WYE354, WYE132, OSI-027, OXA-01, PI-103, NVP-BEZ235, GNE-493, GSK2126458, rapamycin, tacrolimus, everolimus, RAD001, CCI-779 and AP23573.

[0135] In some embodiments of the vaccine for inducing tolerance, the at least one D is covalently linked to the hydrophobic block (H) directly or indirectly via a linker X1.

[0136] In some embodiments of the vaccine for inducing tolerance, the linker X1 comprises an amide, carbamate, hydrazone, ketal, or silyl ether moiety.

[0137] In some specific embodiments of the vaccine for inducing tolerance, the linker X1 comprises a degradable peptide containing 2 to 6 amino acids.

[0138] In some embodiments of the vaccine for inducing tolerance, the linker X1 comprising an enzymatically degradable peptide comprises an amino acid residue P1 selected from the group consisting of arginine, lysine, acetyl lysine, boc-protected lysine, citrulline, glutamine, threonine, leucine, norleucine, α-aminobutyric acid, and methionine; and an amino acid residue P2 selected from the group consisting of β-alanine, glycine, serine, leucine, valine, and isoleucine.

[0139] In a thirteenth aspect of the present disclosure, provided herein is a vaccine comprising at least one peptide antigen conjugate having a formula selected from PEG-[E1]-A-[E2]-[U]-H and H-[U]-[E1]-A-[E2]-PEG, wherein

[0140] E1 is the N-terminal extension;

[0141] E2 is the C-terminal extension;

[0142] H is independently a hydrophobic block at each occurrence, wherein one or more drug molecules (D) are optionally attached to each H directly or via a suitable linker X1;

[0143] U is independently a linker at each occurrence;

[0144] [ ] indicates that the group is optional, and

[0145] - means that two adjacent groups are directly attached to each other by a covalent bond or indirectly attached to each other via a suitable linker X; and

[0146] A is a peptide antigen selected from the group consisting of QLQPFPQPELPYPQPQLPYPQPQPFR (SEQ ID NO:486), PQLPYPQPELPYPQPQPFRPEQPYPQPQP (SEQ ID NO:487), QGIIQPEQPAQLEVI (SEQ ID NO:464), PQPQQPEQPFPQPEQEFPQPQQPQQSFPEQQPPL (SEQ ID NO:488), PQQPFPQPEQPFCQQPQ (SEQ ID NO:489), QQFLQPEQPFPQQPEQPYPQQPEQPFPQPQQ (SEQ ID NO:490), QQFSQPEQEFPQPQQPQQSFPEQQPPF (SEQ ID NO:491), PTPLQPEQPFPQQPQQPQQPFPQPEQPFPWQPQ (SEQ ID NO:492). NO:492), SSPLQPEQPFPQQPQQPFPEQPQQPQ (SEQ ID NO:493), QSIPQPEQPFPQPEQPFPQSQE (SEQ ID NO:494), PQQPFPQQPQQIIPQ (SEQ ID NO:495), PQQPIPEQPQPYPEQPQPYPQQ (SEQ ID NO: 496), QQPPFSEQEQPVLPQ (SEQ ID NO:484), QPPFSQQQESPFSQQ (SEQ ID NO:485), and PQQPFPQPEQPFBQQPQ (SEQ ID NO:497).

[0147] In one embodiment of the vaccine of the present disclosure, the peptide antigen conjugate having a formula selected from PEG-[E1]-A-[E2]-[U]-H and H-[U]-[E1]-A-[E2]-PEG self-assembles into nanoparticle micelles when the peptide antigen conjugate comprises a peptide antigen (A) having a water solubility of at least 1 mg / mL, or wherein the peptide antigen conjugate exhibits an aggregation tendency after 24 h at room temperature when the total peptide antigen conjugate concentration is ≥0.5 mM in an aqueous formulation buffer containing no more than 20% organic solvent.

[0148] In some embodiments of the vaccines of the present disclosure, the micelles have a diameter between about 5 nm and about 50 nm, or a diameter between about 10 nm and about 30 nm.

[0149] In one embodiment of the vaccine of the present disclosure, at least one drug molecule (D) is non-covalently associated with the micelle.

[0150] In a fourteenth aspect of the present disclosure, provided herein is a vaccine formulation comprising the following composition: at least one peptide antigen conjugate having a formula selected from PEG-[E1]-A-[E2]-[U]-H and H-[U]-[E1]-A-[E2]-PEG, wherein

[0151] A is a peptide antigen;

[0152] E1 is the N-terminal extension;

[0153] E2 is the C-terminal extension;

[0154] H is independently a hydrophobic block at each occurrence, wherein one or more drug molecules (D) are optionally attached to each H directly or via a suitable linker X1;

[0155] U is independently a linker at each occurrence;

[0156] [ ] indicates that the group is optional, and

[0157] - means that two adjacent groups are directly attached to each other by a covalent bond or indirectly attached to each other via a suitable linker X

[0158] and a formulation buffer comprising 10% DMSO (v / v) in phosphate-buffered saline (pH 7.4) or tris(hydroxymethyl)aminomethane in saline (0.9% NaCl) (pH 6.5-8.5).

[0159] In one embodiment of the vaccine formulation, a non-ionic surfactant is further included.

[0160] In some embodiments of the vaccine formulation, the nonionic surfactant is selected from polysorbate-20 and sodium lauryl sulfate. BRIEF DESCRIPTION OF THE DRAWINGS

[0161] Figure 1A-1B Shows the turbidity of the vaccine formulation ( Figure 1A ) and particle size ( Figure 1B ), the vaccine formulations comprise peptide antigen conjugates with different N-terminal groups (i.e., charged block (C) or PEG). For a description of materials and methods, see Table 1C and the Experimental section.

[0162] Figure 2 It was shown that replacing the positively charged block (C) of the peptide antigen conjugate of formula C-E1-A-E2-UH with a PEG group to generate a conjugate of formula PEG-E1-A-E2-UH abolished the dose-dependent hemolytic activity associated with the charged block. For a description of materials and methods, see Table 2B and the Experimental section.

[0163] Figure 3A-3C The data show that the N-terminal group of the peptide antigen conjugate has an effect on tolerability after intravenous administration. The data show that the vaccine formulation containing the peptide antigen conjugate of the formula PEG-E1-A-E2-UH is better tolerated than the peptide antigen conjugate of the formula C-E1-A-E2-UH after intravenous administration. Figure 3A Shows the experimental plan and vaccination schedule. Figure 3B Body weight kinetics are shown, with body weight normalized to day 0. Figure 3C Survival rates are shown on Kaplan-Meier curves. For a description of materials and methods, see Table 3B and the Experimental section.

[0164] Figures 4A-4C Shown are the kinetics of disability scores in mice with experimental autoimmune encephalomyelitis (EAE) that received different treatments after EAE disease induction at time 0 (Table 4). The data show that treatment with a vaccine composition comprising a peptide antigen conjugate of the formula PEG-E1-A-E2-UH or C-E1-A-E2-UH reversed the disease, but when administered by the SC route, the peptide antigen conjugate of the formula PEG-E1-A-E2-UH provided superior efficacy compared to the peptide antigen conjugate of the formula C-E1-A-E2-UH. Figure 4A Experimental plan showing the dates of EAE induction and treatment. Figure 4B-4C The kinetics of disability scores are shown for the groups administered by intravenous and subcutaneous routes.

[0165] Figure 5A-5BThe effects of different treatments (Table 4) on the T cell phenotype of mice with experimental autoimmune encephalomyelitis (EAE) are shown. The data show that vaccines containing peptide antigen conjugates of the formula PEG-E1-A-E2-[U]-H and C-E1-A-E2-[U]-H reduced the number of IFN-γ expressing CD4 T cells (Th1 CD4 T cells, Figure 5A ) and IL-17-expressing CD4 T cells (Th17 CD4 T cells, Figure 5B ), but the peptide antigen conjugate of formula PEG-E1-A-E2-UH provided a greater reduction in IFN-γ-producing cells as compared to the peptide antigen conjugate of formula C-E1-A-E2-UH. Asterisks (*) indicate that p < 0.05 for a Student's T-test comparing stimulated samples to unstimulated samples.

[0166] Figures 6A-6D Shown are the kinetics of disability scores in mice with experimental autoimmune encephalomyelitis (EAE) that received different treatments on days 0 and 28 after EAE disease induction (Table 5). Figure 6A Shown is the experimental plan including multiple days of EAE induction and treatment. Figure 6B-6C The kinetics of disability scores are shown for the groups receiving treatment by the intravenous (IV) or intramuscular route (IM). Figure 6D Shown are the kinetics of disability scores in mice treated with IM after the second induction of EAE. The data demonstrate that treatment with a vaccine composition comprising a peptide antigen conjugate of either PEG-E1-A-E2-UH or C-E1-A-E2-UH reversed the disease, but that the peptide antigen conjugate of the formula PEG-E1-A-E2-UH provided superior efficacy compared to the peptide antigen conjugate of the formula C-E1-A-E2-UH, and that PEG-E1-A-E2-UH exhibited comparable efficacy via IM and IV routes.

[0167] Figure 7A-7B The effects of different treatments (Table 5) on the T cell phenotype of mice with experimental autoimmune encephalomyelitis are shown. The data show that vaccines containing peptide antigen conjugates of the formula PEG-E1-A-E2-[U]-H and C-E1-A-E2-[U]-H reduced the number of IFN-γ expressing CD4 T cells (Th1 CD4 T cells, Figure 7A ) and IL-17-expressing CD4 T cells (Th17 CD4 T cells, Figure 7B), and the vaccine containing Torin resulted in a decrease in the proportion of CD4 T cells expressing IFN-γ compared to animals treated with the vaccine containing rapamycin (Group 7). Asterisks (*) indicate Student's T-test comparing stimulated samples to unstimulated samples, p < 0.05.

[0168] Figure 8A-8B Shown are the turbidity at 1 hour (h) and 24 hours (h) for representative vaccines (Table 8) at varying storage temperatures and peptide antigen conjugate concentrations. Figure 8B Filtration recoveries for representative vaccines (Table 8) at varying storage temperatures and peptide antigen conjugate concentrations are shown. Filtration recoveries were assessed 24 hours (h) after formulation and storage by filtering each vaccine composition through a 0.2 µm filter and then assessing the area under the curve by HPLC at 220 nm, dividing the pre-filtration AUC by the post-filtration AUC.

[0169] Figure 9A Shown are the turbidity at 1 hour (h) and 24 hours (h) for representative vaccines (Table 10) with varying peptide antigen conjugate (PAC) formula, surfactant identity and concentration, presence of amphiphile, and formulation method. Figure 9B Shown are 24 hour (h) filtration recoveries for representative vaccines (Table 10) with varying PAC formula, surfactant identity and concentration, presence of amphiphile, and formulation method.

[0170] Figure 10A Shown are the turbidity at room temperature (approximately 23° C.) for 1 hour, 24 hours, and 48 hours for representative vaccines (Table 13) with varying PAC formula, surfactant identity, presence of amphiphile, and formulation method. Figure 10B Shown are 24 and 48 hour filtration recoveries at room temperature (approximately 23°C) for representative vaccines (Table 13) with varying PAC formula, emulsifier identity, presence of amphiphile, and formulation method.

[0171] Figure 11A Shown are the turbidity of representative vaccines (Table 14) at room temperature (approximately 23°C) at 24 and 48 hours with varying mTORi identity, mTORi molar ratio, PAC concentration, and formulation buffer. Figure 11B Shown are filtration recoveries of representative vaccines (Table 14) at room temperature (approximately 23°C) for 1 hour, 24 hours, and 48 hours, under varying mTORi identities, mTORi molar ratios, PAC concentrations, and formulation buffers.

[0172] Figure 12A Shown are the turbidity of representative vaccines (Table 15) at room temperature (approximately 23°C) at 24 and 48 hours with varying mTORi identities, mTORi molar ratios, and PAC concentrations. Figure 12B Shown are filtration recoveries of representative vaccines (Table 15) at room temperature (approximately 23°C) for 1 hour, 24 hours, and 48 hours at varying mTORi identities, mTORi molar ratios, and PAC concentrations.

[0173] Figures 13A-13D Shown are the kinetics of disability scores in mice with experimental autoimmune encephalomyelitis (EAE) that received different treatments on day 0, day 7, and day 14 of treatment following EAE disease induction (Table 17). Figure 13A The experimental plan including EAE induction and treatment days is shown. Figures 13B-13D Shown are the kinetics of disability scores for the groups receiving treatment by the intramuscular route (IM). Figure 13B It was shown that inclusion of an irrelevant peptide antigen (CPNE1) did not affect efficacy. Figure 13C showed that inclusion of Torin-1 or rapamycin improved efficacy. Figure 13D The results showed that the inclusion of rapamycin at a 1:1 molar ratio of drug molecule to peptide antigen conjugate showed the highest efficacy compared to the absence of rapamycin or a 0.5:1 molar ratio of rapamycin to peptide antigen conjugate. The data showed that treatment with a vaccine composition comprising a peptide antigen conjugate of PEG-E1-A-E2-UH provided protection from EAE disease, but the peptide antigen conjugate of the formula PEG-E1-A-E2-UH + D provided superior efficacy compared to the same formulation lacking the mTOR inhibitor drug.

[0174] Figures 14A-14F Shown are the kinetics of disability scores in mice with experimental autoimmune encephalomyelitis (EAE) that received different treatments on day 0, day 7, and day 14 of treatment following EAE disease induction (Table 18). Figure 14A The experimental plan including EAE induction and treatment days is shown. Figures 14B-14D The kinetics of disability scores for groups treated with 2.5, 10, or 40 nmol of peptide antigen, respectively, are shown. At equivalent doses, the four formulations tested showed effectively equivalent efficacy. The data show that formulations consisting of PEG-E1-A-E2-UH were equivalent or superior in efficacy to formulations consisting of (i) E1-A-E2-UH + SB-[U]-H, (ii) PEG-E1-A-E2-UH + SB-[U]-H + surfactant, or (iii) PEG-E1-A-E2-UH + SB-[U]-H. Figure 14E Shown is a comparison of EAE disease score efficacy for PEG-E1-A-E2-UH formulations at doses of 2.5, 10, or 40 nmol, respectively. Figure 14F Shown are estimates of the area under the curve for EAE disease scores from day 7 to day 28 using between-group statistical evaluation (one-way ANOVA with Tukey correction for multiple comparisons, where asterisks indicate p-values ​​≤ 0.0001). The data demonstrate a strong correlation between the dose of peptide antigen construct administered and EAE disease score.

[0175] Figures 15A-15C Shown are the disability score kinetics of mice with experimental autoimmune encephalomyelitis (EAE) and the area under the disease score curve (AUC) of mice that received different treatments on day 0, day 7, and day 14 of treatment following EAE disease induction (Table 19). Figure 15A The experimental plan including EAE induction and treatment days is shown. Groups

[12] and

[13] received treatment only on day 0. Figure 15B Shown is a comparison of efficacy on EAE disease scores in mice dosed IM or SC, given three treatments or a single treatment on day 0 only. Figure 15C Shown are estimates of the area under the curve for EAE disease scores from day 7 to day 21.

[0176] Figures 16A-16G Shown are the experimental plan and immunological outcomes for testing GLU peptide antigen conjugates using formulations (Table 21) in Sprague Dawley rats. Figure 16A The experimental plan is shown, which includes gliadin and gluten peptide sensitization on days -14 and -7, followed by peptide antigen conjugate treatment on days 0, 7, 14, 21 and 28. Figure 16B Shown are anti-gliadin antibody titers in both gliadin-sensitized and non-sensitized (naive) rats on day 35 of the study. Figure 16C Shown are assessments of phospho-S6 activation levels in antigen presenting cells (RT1B+, CD3-) isolated from blood four hours after treatment on study day 0. Figure 16D The frequency (%) of IFN-γ+ cells in the CD4+ cell population isolated from blood on day 35 is shown. Figure 16E The frequency (%) of IL-17+ cells in the CD4+ cell population isolated from blood on day 35 is shown. Figure 16F The frequency (%) of IFN-γ+ cells in the CD8+ cell population isolated from blood on day 35 is shown. Figure 16G The frequency (%) of IL-17+ cells in the CD8+ cell population isolated from blood on day 35 is shown.

[0177] Figures 17A-17CThe results of the in vitro mTOR inhibitor assays in splenocyte groups are shown after in vitro stimulation with LPS. Splenocytes from C57BL / 6 mice are dissociated into single cell suspensions and plated in vitro for stimulation. Test compound is added to cells at 0.5, 5, 50 or 500 nM concentrations and kept for one hour. Thereafter, lipopolysaccharide (LPS) is added to cells and kept for two hours to stimulate the phosphorylation of mTOR activity and downstream S6 to phospho-S6 (pS6). Cells are then fixed and stained for flow cytometry analysis. Before being added to splenocyte cultures, test compound (Table 24) is resuspended in PBS. Figure 17A Demonstrating mTOR inhibition as measured by the pS6 assay using a small molecule drug analog of everolimus with a conjugatable handle. Figure 17B Demonstration of mTOR inhibition as measured by the pS6 assay using peptide antigen conjugates with everolimus derivatives conjugated to an H block. These two examples show that everolimus or rapamycin can be modified for conjugation to other molecules and that the conjugated forms of the mTOR inhibitors retain activity when conjugated via an amide linkage (group [7]) or a cleavable linkage (group [8]). Figure 17C Showing mTOR inhibition as measured by the pS6 assay using a small molecule analog of Torin-1. DETAILED DESCRIPTION

[0178] The above-mentioned aspects and other aspects, features and advantages of the present disclosure are described below in conjunction with various embodiments with reference to the accompanying drawings.

[0179] definition

[0180] For the purpose of guiding those skilled in the art in the practice of this disclosure, the details of terms and methods are given below to make people more aware of compounds, compositions, methods and one or more uses thereof. The terms in this disclosure should be understood to be used for the purpose of better describing specific embodiments and should not be regarded as limiting.

[0181] About: In the context of the present disclosure, "about" when referring to a measurable value such as an amount, duration, etc., is intended to encompass variations of ±20%, ±10%, ±5%, ±1%, or ±0.1% of the specified value, as such variations are appropriate for performing the disclosed methods. For example, "about 10" refers to 9.5 to 10.5. A ratio of "about 5:1" refers to a ratio of 4.75:1 to 5.25:1.

[0182] Administration: providing or administering an agent, e.g., an immunogenic composition comprising an amphiphilic block copolymer and one or more drugs as described herein, to a subject by any effective route. Exemplary routes of administration include, but are not limited to, oral, injection (e.g., subcutaneous, intramuscular, intradermal, intraperitoneal, and intravenous), transdermal, topical, intranasal, vaginal, and inhalation routes.

[0183] "Administration" of a compound and "administering" a compound should be understood to mean providing a compound, a prodrug of a compound, or a pharmaceutical composition as described herein. The compound or composition can be administered to the subject by another person or can be self-administered by the subject.

[0184] Antigen: any molecule containing an immune response (particularly a B cell response and / or a T cell response) that binds to a T cell or B cell receptor and can stimulate a subject's immune response. An epitope can include a peptide, glycopeptide, lipid or any suitable molecule containing an epitope that can interact with a component of a specific B cell or T cell receptor. Such interactions can produce a response of an immune cell. An "epitope" refers to a region in a peptide antigen that interacts with a B and / or T cell protein (i.e., a B cell receptor and a T cell receptor). The antigen used in the embodiments of the present disclosure can be selected from pathogens, cancer cells, autoantigens, alloantigens or allergens. Many such antigens can be used according to embodiments of the present disclosure and are discussed in more detail throughout this specification.

[0185] Antigen presenting cell (APC): Any cell that presents antigen bound to MHC class I or class II molecules to T cells (including but not limited to monocytes, macrophages, dendritic cells, B cells, T cells, and Langerhans cells).

[0186] Amphiphilic: The term "amphiphilic" is used herein to refer to a substance that contains both hydrophilic or polar groups and hydrophobic groups.

[0187] CD4: Cluster of differentiation 4, a surface glycoprotein that interacts with MHC class II molecules present on the surface of other cells. A subset of T cells express CD4, and these cells are generally referred to as helper T cells or CD4 T cells.

[0188] CD8: Cluster of differentiation 8, a surface glycoprotein that interacts with MHC class I molecules present on the surface of other cells. A subset of T cells express CD8, and these cells are generally referred to as cytotoxic T cells (CTLs), killer T cells, or CD8 T cells.

[0189] Charge: A physical property of a substance that affects its interactions with other atoms and molecules, including solutes and solvents. Charged substances experience electrostatic forces from other types of charged substances and molecules that do not hold a full integer value of charge, such as polar molecules. Two charged molecules with the same charge repel each other, while two charged molecules with different charges attract each other. Charge is typically described in positive or negative integer units. The charge of a molecule can be easily estimated based on the Lewis structure of the molecule and recognized methods known to those skilled in the art. Charge may result from inductive effects, for example, atoms with different electron affinities when bonded together may produce polar covalent bonds, resulting in partially negatively charged atoms and partially positively charged atoms. For example, nitrogen bonded to hydrogen results in a partial negative charge on the nitrogen and a partial positive charge on the hydrogen atom. Alternatively, an atom in a molecule can be considered to have a full integer value of charge when the number of electrons assigned to it is less than or equal to the atomic number of the atom. The charge of a molecule is determined by summing the charges of each atom that makes up the molecule. Those skilled in the art are familiar with the process of estimating the charge of a molecule by summing the formal charges of each atom in the molecule. "A charged functional group refers to a functional group that can be permanently charged or charged depending on the pH. A charged functional group can have a partial or full integer value of charge (the charge can be positive or negative), which functional group is referred to as a positively charged functional group or a negatively charged functional group, respectively. A portion of a molecule that contains one or more charged functional groups (which can be positive or negative) is referred to as a "charged group," e.g., a positively charged group or a negatively charged group. A charged group can include a positive functional group, a negative functional group, or both positive and negative functional groups. The net charge of a charged group can be positive, negative, or neutral. A charged monomer refers to a monomer that contains a charged group. Charged amino acids are one type of charged monomer. Note: The net charge of a particle containing an amphiphile and / or peptide antigen conjugate that further contains a charged group, e.g., a charged monomer such as a charged amino acid, can be estimated by summing the charges of each functional group within the amphiphile and / or peptide antigen conjugate. A charged block (C) is a type of solubilizing block.

[0190] Click chemistry: A bioorthogonal reaction that joins two compounds together under mild conditions in a high-yield reaction with minimal, biocompatible, and / or harmless byproducts. An exemplary click chemistry reaction used in this disclosure is the reaction of an azide group with an alkyne to form a triazole linker via a strain-promoted [3+2] azide-alkyne cycloaddition.

[0191] Copolymer: A polymer derived from two (or more) different monomers, as opposed to a homopolymer, which uses only one monomer. Because copolymers include at least two types of constituent units (also structural units), they can be classified based on how these units are arranged along the chain. Copolymers can be statistical (or random) copolymers, in which the two or more monomer units are randomly distributed; copolymers can be alternating copolymers, in which the two or more monomer units are distributed in an alternating order; or, for example, copolymers such as poly(amino acids) can be produced by solid phase peptide synthesis (SPPS) and have a specific order of monomer units. The term "block copolymer" generally refers to a polymer composed of two or more adjacent blocks of different constituent monomers or comonomers (if the block contains two or more different monomers). Block copolymers can be used herein to refer to copolymers comprising two or more homopolymer subunits, two or more copolymer subunits, or one or more homopolymer subunits and one or more copolymer subunits, wherein the subunits can be directly linked by covalent bonds, or the subunits can be indirectly linked via intermediate non-repeating subunits (such as connecting blocks or linkers). Blocks can be based on linear and / or brush-like architectures. Block copolymers having two or three different blocks are referred to herein as "diblock copolymers" and "triblock copolymers," respectively. Copolymers may be generally referred to as polymers, for example, statistical copolymers may be referred to as polymers or copolymers. Similarly, block copolymers may be generally referred to as polymers. While copolymers as used herein mean polymers comprising two or more types of monomers, terpolymers are copolymers having three monomer units.

[0192] Critical Micelle Concentration (CMC): refers to the concentration of a material above which micelles spontaneously form to satisfy thermodynamic equilibrium.

[0193] Drug: refers to any pharmaceutically active molecule, including but not limited to proteins, peptides, carbohydrates, sugars, nucleosides, inorganic compounds, lipids, nucleic acids, small synthetic chemical compounds, macrocyclic compounds, etc., which has a physiological effect when ingested or otherwise introduced into the body. Pharmaceutically active compounds can be selected from a variety of known classes of compounds, including, for example, analgesics, anesthetics, anti-inflammatory agents, anthelmintics, antiarrhythmics, antiasthmatics, antibiotics (including penicillins), anticancer agents, anticoagulants, antidepressants, antidiabetics, antiepileptics, antihistamines, antitussives, antihypertensives, antimuscarinics, antimycobacterials, antitumor agents, antioxidants, antipyretics, immunosuppressants, immunostimulants, antithyroid agents, antivirals, antianxiety sedatives (hypnotics and neuroleptics), astringents, antibacterials, beta-adrenergic receptor blockers, blood products and substitutes, bronchodilators, buffers, myocardial inotropes, chemotherapeutic agents , contrast media, corticosteroids, antitussives (expectorants and mucolytics), diagnostic agents, diagnostic imaging agents, diuretics, dopamine mimetics (antiparkinsonian agents), free radical scavengers, growth factors, hemostatics, immune agents, lipid regulators, muscle relaxants, proteins (such as therapeutic antibodies and antibody fragments, MHC-peptide complexes, cytokines and growth factors, glycoproteins, peptides and polypeptides), parasympathomimetics, parathyroid calcitonin, bisphosphonates, prostaglandins, radiopharmaceuticals, hormones, sex hormones (including steroids), antiallergic agents, stimulants and anorexigens, steroids, sympathomimetics, thyroid agents, vaccines, vasodilators and xanthines. Drugs can also be referred to as pharmaceutically active agents, pharmaceutically active substances or biologically active compounds or biologically active molecules. Any drug molecule in the formula described herein is abbreviated as "D".

[0194] Drug delivery: The method or process of administering a pharmaceutical compound to achieve a therapeutic effect in humans or animals.

[0195] Effective amount: An amount of a compound, material, or composition effective to achieve a specific biological result (such as, but not limited to, those disclosed, described, or exemplified herein). Such results may include, but are not limited to, an effective reduction in symptoms associated with any disease state mentioned herein, as determined by any suitable means in the art.

[0196] Hydrophilicity index / GRAVY value: It is a number representing the hydrophobic or hydrophilic characteristics of an amino acid or amino acid sequence. There are a variety of scales that can be used to describe the relative hydrophobic and hydrophilic characteristics of the amino acids that make up a peptide. In the present disclosure, the hydrophilicity scale of Kyte and Doolittle (Kyte J, Doolittle RF, J. Mol. Biol 157: 105–32, 1983) is used to calculate the overall average hydrophilicity (GRAVY) value (sometimes referred to as the GRAVY score). The GRAVY value of a peptide is the sum of the hydrophilicity values ​​of all the amino acids that make up the peptide divided by the length of the peptide (i.e., the number of amino acids). The GRAVY value is a relative value. The larger the GRAVY value, the more hydrophobic the peptide sequence is, and the lower the GRAVY value, the more hydrophilic the peptide sequence is.

[0197] Hydrophilic: refers to the tendency of a material to disperse freely or be dissolved in an aqueous solution (sometimes referred to as an aqueous medium). A material is considered hydrophilic if it prefers to interact with other hydrophilic materials and avoids interacting with hydrophobic materials. In some cases, hydrophilicity can be used as a relative term, for example, the same molecule can be described as hydrophilic or non-hydrophilic, depending on what it is being compared to. Hydrophilic molecules are generally polar and / or charged and have good water solubility, for example, soluble at a concentration of at least 1.0 mg / mL or more. A hydrophilic group refers to a portion of a molecule that is polar and / or charged and has good water solubility.

[0198] Hydrophobic: refers to the tendency of a material to avoid contact with water. A material is considered hydrophobic if it prefers to interact with other hydrophobic materials and avoids interacting with hydrophilic materials. Hydrophobicity is a relative term; the same molecule can be described as hydrophobic or non-hydrophobic, depending on what it is compared with. Hydrophobic molecules are typically non-polar and uncharged and have poor water solubility, for example, insoluble in water, or soluble in water only at a concentration of less than 1 mg / mL, typically 0.1 mg / mL or lower, or more preferably 0.01 mg / mL or lower. A hydrophobic monomer is a monomer (e.g., a hydrophobic amino acid) that contains a hydrophobic group and forms a polymer that is insoluble in water or insoluble in water at certain temperatures, pH, and salt concentrations. A hydrophobic group refers to the hydrophobic portion of a molecule. For example, a styrene monomer can be referred to as a hydrophobic monomer because poly(styrene) is a water-insoluble polymer. Hydrophobic drugs refer to drug molecules that are insoluble in aqueous solution at a pH of about pH 7.4 or are soluble only at a concentration of about 1.0 mg / mL or less. Amphiphilic drugs are drug molecules that have a tendency to assemble into supramolecular structures (e.g., micelles) in aqueous solution and / or have limited solubility in aqueous solution at a pH of about pH 7.4.

[0199] Immune response: A change in the activity of cells of the immune system (such as B cells, T cells, or monocytes) due to a stimulus, either directly or indirectly (such as through cell or cytokine mediation). In certain embodiments, the response is specific for a particular antigen ("antigen-specific response"). The immune response may include a T cell response, such as a CD4 T cell response or a CD8 T cell response. This immune response may result in the production of additional T cell progeny and / or the migration of T cells. In other embodiments, the response is a B cell response and results in the production of specific antibodies or the production of additional B cell progeny. In yet other embodiments, the response is an antigen-presenting cell response. Antigens can be used to stimulate an immune response, resulting in the activation of cytotoxic T cells that kill virus-infected cells or cancer cells. In other embodiments, antigens can be used to induce tolerance or immunosuppression. Tolerogenic responses may result from the unresponsiveness of T cells or B cells to antigens. Suppressive immune responses may result from the priming and / or activation of regulatory cells, such as regulatory T cells, or the transdifferentiation of effector cells into regulatory cells, thereby downregulating the immune response, i.e., weakening the immune response.

[0200] Immunogenic composition: A formulation of materials comprising an antigen and, optionally, an immunomodulatory agent that induces a measurable immune response against the antigen. For example, vaccines are one type of immunogenic composition.

[0201] Immunomodulators: refers to a class of drugs that regulate the activity of cells of the immune system, including immunostimulants and immunosuppressants.

[0202] Immunostimulant: refers to any synthetic or naturally occurring drug that promotes the pro-inflammatory and / or cytotoxic activity of immune cells. Exemplary immunostimulants include pattern recognition receptor (PRR) agonists, such as synthetic or naturally occurring agonists of Toll-like receptors (TLRs), interferon gene stimulator agonists (STINGa), nucleotide binding oligomerization domain-like receptors (NLR) agonists, retinoic acid-inducible gene-I-like receptors (RLR) agonists and certain C-type lectin receptors (CLRs), and certain cytokines (e.g., certain interleukins, such as IL-2); certain chemokines or small molecules that bind to chemokine receptors; certain antibodies, antibody fragments or synthetic peptides that activate immune cells, for example, by binding to stimulatory receptors, for example, anti-CD40, or for example by blocking inhibitory receptors, for example, anti-CTLA4, anti-PD1, etc. Various immunostimulants suitable for the practice of the present disclosure are described throughout this specification. For the sake of clarity, certain pharmaceutically active compounds that stimulate the immune system may be referred to as immunostimulants, or more generally as drug molecules (abbreviated "D" in the formula).

[0203] Immunosuppressants: refers to any synthetic or naturally occurring drug that inhibits the proinflammatory and / or cytotoxic activity of immune cells or the humoral immune system (e.g., antibodies and complement proteins). Immunosuppressants can mediate effects via one or more of the following mechanisms of action: by activating suppressor cells, e.g., regulatory T cells; killing, inhibiting, or inactivating proinflammatory cells, cytotoxic cells, and / or B cells; transdifferentiating proinflammatory and / or cytotoxic T cells into suppressor cells; and / or isolating proinflammatory cells, cytotoxic cells, and / or B cells and / or limiting the motility of the cells. Exemplary immunosuppressants include synthetic or naturally occurring agonists of aryl hydrocarbon receptor (AHR); certain steroids, including glucocorticoids; certain histone deacetylase inhibitors (HDACS), such as inhibitors of HDAC9; retinoic acid receptor agonists; mammalian target of rapamycin (mTOR) inhibitors, such as rapamycin; certain cyclin-dependent kinase (CDK) inhibitors; certain adenosine receptor agonists; agonists of PD1; and other molecules that suppress the proinflammatory or cytotoxic activity of immune cells or antibodies. Various immunosuppressants suitable for the practice of the present disclosure are described throughout this specification, and include immunomodulators that promote Treg. For clarity, immunosuppressants can be more generally referred to as drug molecules (abbreviated as "D" in the formula).

[0204] In vivo delivery: A composition (e.g., a composition comprising an amphiphilic block copolymer and one or more drugs) is administered to a subject topically, transdermally, by suppository (rectal, vaginal), pessary (vaginal), intravenously, orally, subcutaneously, intraperitoneally, intrathecally, intramuscularly, intracranially, by inhalation, orally, or by any other suitable route.

[0205] Connect or couple: The terms "connect" and "couple" mean directly or indirectly joined together. The first part can be covalently or non-covalently linked to the second part. In some embodiments, the first molecule is connected to another molecule by a covalent bond. In some embodiments, the first molecule is connected to another molecule by electrostatic attraction. In some embodiments, the first molecule is connected to another molecule by dipole-dipole forces (e.g., hydrogen bonding). In some embodiments, the first molecule is connected to another molecule by van der Waals forces (also known as London forces). The first molecule can be connected to another molecule by any one and all combinations of such couplings. The molecules can be indirectly connected, such as by using a linker (sometimes referred to as a linker molecule). The molecules can be indirectly connected by the insertion of components that are independently non-covalently bound to the two molecules. The term "linker" (sometimes abbreviated as "X") used in the chemical formula herein means any suitable linker molecule. Particularly preferred linkers can be indicated by other symbols, such as X1, X2, X3, X4, X5 and U. Various linkers are described throughout this specification.

[0206] A "bilayer membrane" or "bilayer(s)" is a membrane of amphiphiles or superamphiphiles self-assembled in aqueous solution.

[0207] Micelles: Spherical containers with a single monolayer defining a closed compartment. Typically, amphiphilic molecules spontaneously form micellar structures in polar solvents. In contrast to bilayers such as liposomes, micelles are "sided" in that they have a prominent hydrophilic polar outer surface and exhibit a hydrophobic inner surface.

[0208] Mol%: refers to the percentage of a particular type of monomer unit (or "monomer") present in a polymer. For example, a polymer having 100 monomer units of A and B with a density (or "mol%") of monomer A equal to 10 mol% would have 10 monomer units of A, and the remaining 90 monomer units (or "monomers") could be monomer B or another monomer, unless otherwise specified.

[0209] Monomeric unit: The term "monomeric unit" or "monomer unit" is used herein to refer to a unit in a polymer molecule that contains the same or a similar number of atoms as one of the monomers. Monomeric units, as used in this specification, can be of a single type (homogeneous) or multiple types (heterogeneous). For example, a poly(amino acid) comprises amino acid monomer units. A monomeric unit may also be referred to as a monomer or monomer unit, etc.

[0210] Net Charge: The sum of the electrostatic charges carried by a molecule or (if specified) parts or segments of a molecule.

[0211] Particle: A nanoscale or micrometer-scale supramolecular structure composed of an assembly of molecules. For example, the amphiphiles and peptide antigen conjugates of the present disclosure form particles in an aqueous solution. In some embodiments, the particle formation of the amphiphiles and / or peptide antigen conjugates is dependent on pH or temperature. In some embodiments, the average diameter of the nanoparticles composed of the amphiphiles and / or peptide antigen conjugates is between 5 nanometers (nm) and 500 nm. In some embodiments, the nanoparticles composed of the amphiphiles and / or peptide antigen conjugates form micelles, and the average diameter is between 5 nanometers (nm) and 50 nm, such as between 10 nm and 30 nm. In some embodiments, the nanoparticles composed of the amphiphiles and / or peptide antigen conjugates may be larger than 100 nm.

[0212] Pattern recognition receptors (PRRs): receptors expressed by various cell populations (particularly innate immune cells) that bind to a variety of synthetic and naturally occurring molecules. There are several classes of PRRs. Non-limiting examples of PRRs include Toll-like receptors (TLRs), RIG-I-like receptors (RLRs), NOD-like receptors (NLRs), stimulator of interferon genes receptors (STINGs), and C-type lectin receptors (CLRs). Agonists of such PRRs are referred to as immunostimulant drugs and can be used to enhance and / or modify immune responses to antigens. For more information on pattern recognition receptors, see Wales et al., Biochem Soc Trans., 35: 1501-1503, 2007.

[0213] Peptide or polypeptide: two or more natural or non-natural amino acid residues joined together in series via one or more amide bonds. The amino acid residues may contain one or more post-translational modifications (e.g., glycosylation, citrullination, homocitrullination, oxidation and / or phosphorylation). Such modifications may mimic post-translational modifications that occur naturally in vivo or may be non-natural. Any one or more components of the amphiphile and / or peptide antigen conjugate may comprise a peptide.

[0214] Peptide modifications: Peptides can be altered or otherwise synthesized to have one or more of several modifications as described below. In addition, analogs (non-peptide organic molecules), derivatives (chemically functionalized peptide molecules obtained starting from the peptides) and variants (homologs) of these peptides can be utilized in the methods described herein. The peptides described herein comprise sequences of amino acids, analogs, derivatives and variants (which may be in L- and / or D- form). Unless otherwise specified, any peptide sequence mentioned herein comprises L amino acids, preferably only L amino acids. Such peptides may contain peptides, analogs, derivatives and variants of naturally occurring and other origins.

[0215] Peptides can be modified by any of a variety of chemical techniques to produce derivatives having similar activity to the unmodified peptide and optionally other desired properties. For example, the carboxylic acid group of the peptide (whether at the carboxyl terminus or at the side chain) can be provided in the form of a salt of a pharmaceutically acceptable cation or esterified to form a CC1-CC 16 Ester, where CC refers to a carbon chain (and thus, CC1 refers to a single carbon and CC16 refers to 16 carbons), or converted to an amide. The amino group of the peptide (whether at the amino terminus or at the side chain) can be in the form of a pharmaceutically acceptable acid addition salt, such as HCl salt, HBr salt, acetate, trifluoroacetate, formate, benzoate, tosylate, maleate, tartrate and other organic salts, or can be modified or converted to an amide, for example, by acetylation.

[0216] The peptides may be modified to include substituents that contain positive or negative charges or both. The positive and / or negative charges may be affected by the pH at which the peptide is exposed.

[0217] The hydroxyl groups of the peptide side chains can be converted to C1-C 16 Alkoxy or C1-C 16 The esters, or the hydroxyl groups can be converted (e.g., sulfated or phosphorylated) to introduce negative charges. The phenyl and phenol rings of the peptide side chains can be substituted with one or more halogen atoms such as fluorine, chlorine, bromine or iodine, or with C1-C 16 Alkyl, C1-C 16 The alkyl group of the present invention can be replaced by an alkyl group, a carboxylic acid and an ester thereof, or an amide of such a carboxylic acid. The methylene group of the peptide side chain can be extended to a homologous C2-C4 alkylene group. Thiols can be used to form disulfide bonds or thioethers, for example, by reacting with maleimide. Thiols can be protected with any of a variety of recognized protecting groups (such as acetamide groups). Those skilled in the art will also recognize that a method for introducing a cyclic structure into the peptide of the present invention to select and provide a conformational constraint on the structure that results in enhanced stability can be found in Greene et al., "Greene's Protective Groups in Organic Synthesis" Fourth Edition, John Wiley & Sons, Inc. 2006 for details of the additional modifications that can be performed on the functional group.

[0218] The cysteine ​​residues of naturally occurring peptide antigens can be replaced by α-aminobutyric acid or serine, and the methionine residues can be replaced by norleucine to produce non-natural peptide antigens that induce an immune response that cross-reacts with the naturally occurring peptide antigens. Preferred methods for preparing and using peptide antigens with non-natural sequences are described throughout this specification and in WO 2022 / 177993 (incorporated herein by reference).

[0219] Pharmaceutically acceptable vehicles: Pharmaceutically acceptable vehicles (or carriers) useful in the present disclosure include conventional carriers, excipients, and diluents. Remington's Pharmaceutical Sciences, E.W. Martin, Mack Publishing Co., Easton, Pennsylvania, 15th edition (1975) describes compositions and formulations suitable for drug delivery of one or more therapeutic compositions (such as one or more therapeutic cancer vaccines) and additional pharmaceutical preparations.

[0220] Pharmaceutically acceptable carriers are well known in the art and include, for example, aqueous solutions such as water or physiologically buffered saline; or other solvents or vehicles such as glycols, glycerol, oils such as olive oil or injectable organic esters. In a preferred embodiment, when such pharmaceutical compositions are used for human administration, particularly for invasive routes of administration (i.e., routes of transport or diffusion through epithelial barriers, such as injection or implantation), the aqueous solution is pyrogen-free or substantially pyrogen-free. The excipient can be selected to, for example, achieve delayed release of the agent or selectively target one or more cells, tissues or organs. The pharmaceutical composition can be in dosage unit form, such as tablets, capsules (including sprinkle capsules and gelatin capsules), granules, lyophiles for reconstruction, powders, solutions, syrups, suppositories, injections, etc. The composition can also be present in a transdermal delivery system (e.g., a skin patch). The composition can also be present in a solution suitable for topical administration, such as an ointment or cream.

[0221] A pharmaceutically acceptable carrier may contain a physiologically acceptable agent that acts, for example, to stabilize a compound (such as a compound of the present invention), improve the solubility of the compound, or increase the absorption of the compound. Such physiologically acceptable agents include, for example, carbohydrates such as glucose, sucrose, or dextran; antioxidants such as ascorbic acid or glutathione; chelating agents; low molecular weight proteins; or other stabilizers or excipients. The choice of a pharmaceutically acceptable carrier, including a physiologically acceptable agent, depends, for example, on the route of administration of the composition. The preparation of the pharmaceutical composition may be a self-emulsifying drug delivery system or a self-microemulsifying drug delivery system. The pharmaceutical composition (preparation) may also be a liposome or other polymer matrix into which, for example, a compound of the present invention may be incorporated. Liposomes (e.g., comprising phospholipids or other lipids) are non-toxic, physiologically acceptable, and metabolizable carriers that are relatively simple to prepare and administer.

[0222] Polarity: A description of the properties of a substance. Polarity is a relative term and can describe a molecule or portion of a molecule that has a partial charge resulting from differences in electronegativity between atoms bonded together in the molecule, such as the bond between nitrogen and hydrogen. Polar molecules prefer to interact with other polar molecules and generally do not associate with non-polar molecules. In certain non-limiting cases, a polar group can contain a hydroxyl group, an amino group, a carboxyl group, or a charged group. In certain non-limiting cases, a polar group may prefer to interact with polar solvents such as water. In certain non-limiting cases, the introduction of additional polar groups can increase the solubility of a portion of a molecule.

[0223] Polymer: A molecule containing repeating structural units (monomers). As described in more detail throughout this disclosure, polymers can be used for any number of components of amphiphiles, peptide antigen conjugates, and drug molecule conjugates, and can be natural or synthetic. Various compositions of polymers useful in the practice of the present invention are discussed in more detail elsewhere. Note: Polymer is used throughout this specification to broadly encompass molecules having as few as three or more monomers, which molecules may sometimes be referred to as oligomers.

[0224] Polymerization: A chemical reaction, usually carried out with a catalyst, heat or light, in which monomers combine to form chain-like, branched or cross-linked macromolecules (polymers). The chain, branched or cross-linked macromolecules can be further modified by additional chemical synthesis using appropriate substituents and chemical reactions. Polymerization is generally carried out by addition or condensation. Addition polymerization occurs when an initiator (usually a free radical) reacts with a double bond in a monomer. The free radical adds to one side of the double bond, thereby generating a free electron on the other side. The free electron then reacts with another monomer, and the chain begins to self-propagate, thereby adding one monomer unit at a time to the end of the growing chain. Condensation polymerization involves the reaction of two monomer units, resulting in the splitting of a water molecule. In other forms of polymerization, one monomer is added to the growing chain at a time by the staged introduction of activated monomers, such as during solid phase peptide synthesis (SPPS).

[0225] Polymersomes: Vesicles assembled from synthetic multiblock polymers in aqueous solution. Unlike liposomes, polymersomes do not include lipids or phospholipids as their main components. Therefore, compared to the most stable of lipid vesicles, polymersomes may be different in thermal, mechanical and chemical aspects and are particularly more durable and more elastic. Polymersomes assemble during a lamellar swelling process, for example, by film or bulk rehydration or by an additional particle transfer (phoresis) step, as described below, or by other known methods. Like liposomes, polymersomes are formed by "self-assembly", a spontaneous entropy-driven process that prepares a closed semipermeable membrane.

[0226] Purified: A substance or composition that is relatively free of impurities or substances that adulterate or contaminate the substance or composition. The term purified is a relative term and does not require absolute purity. Substantially purified means purified from impurities. A substantially purified substance or composition is typically at least 60%, 70%, 80%, 90%, 95%, 98%, or 99% pure.

[0227] Soluble: Capable of becoming molecularly or ionically dispersed in a solvent to form a homogenous solution. When referring to amphiphiles, peptide antigen conjugates, drug molecule conjugates, and / or drug molecules, soluble is understood to mean a single molecule in solution that does not assemble into multimers or other supramolecular structures via hydrophobic or other non-covalent interactions. A soluble molecule is understood to be freely dispersible in solution as a single molecule. The hydrophobic blocks (H) described herein are insoluble or only soluble to a concentration of about 0.1 mg / mL or less. Solubility can be determined by visual inspection, turbidity measurement, or dynamic light scattering.

[0228] Subject and patient: These terms are used interchangeably herein to refer to both human and non-human animals, including birds and non-human mammals, such as rodents (e.g., mice and rats), non-human primates (e.g., rhesus macaques), companion animals (e.g., domestic dogs and cats), livestock (e.g., pigs, sheep, cattle, llamas, and camels), and non-domestic animals (e.g., big cats).

[0229] Targeting molecules: broadly defined as molecules that guide drug molecules to specific tissues or cell populations. Targeting molecules are defined by their intended use and therefore include structurally diverse molecules, including but not limited to antibodies, Fabs, peptides, aptamers, sugars (e.g., sugars that bind to lectin receptors and / or are recognized by cellular transporters), amino acids, neurotransmitters, and the like. Because targeting molecules are typically selected from molecules that bind to cell receptors that can activate downstream signaling cascades and / or affect the activity of other associated molecules, targeting molecules are generally classified as drug molecules (D) in the present disclosure. In addition, targeting molecules can also have a solubilizing effect and can be regarded as either or both of drug molecules (D) and / or solubilizing groups (SG).

[0230] T cells: a type of white blood cell that is part of the immune system and can participate in the immune response. T cells include but are not limited to CD4 T cells and CD8 T cells. CD4 T cells display CD4 glycoprotein on their surface, and these cells are generally referred to as helper T cells. These cells generally cooperate with the immune response, including antibody response and cytotoxic T cell response, however, CD4 T cells (e.g., regulatory T cells) can also suppress the immune response, or CD4 T cells can function as cytotoxic T cells. CD8 T cells display CD8 glycoprotein on their surface, and these cells are generally referred to as cytotoxic or killer T cells, however, CD8 T cells can also suppress the immune response.

[0231] Treat, prevent, or ameliorate a disease: "Treating" refers to reducing the signs, symptoms, or markers of a disease or pathological condition after it has already begun to develop. For example, treating a disease can result in a reduction in tumor burden, meaning a decrease in the number or size of tumors and / or metastases, or treating a disease can result in immune tolerance, thereby reducing the systemic involvement of autoimmunity. "Preventing" a disease means inhibiting the overall development of the disease. The onset of the disease can be prevented entirely. The severity, extent, or type of the disease can be prevented from developing. "Ameliorging" refers to reducing the number or severity of the signs, symptoms, or markers of a disease, such as cancer.

[0232] Reducing the signs or symptoms or markers of a disease or a pathological state associated with the disease refers to any observable beneficial effect of the treatment and / or any observable effect on a neighboring surrogate endpoint (e.g., tumor volume, whether or not symptomatic). Reducing the signs or symptoms associated with a tumor or viral infection can be demonstrated, for example, by a delayed onset of clinical symptoms of the disease in susceptible subjects (e.g., subjects with tumors that have not yet metastasized or subjects who may be exposed to viral infections), a reduction in the severity of some or all of the clinical symptoms of the disease, a slowing of the progression of the disease (e.g., by extending the life of subjects with tumors or viral infections), a reduction in the number of recurrences of the disease, an improvement in the overall health or well-being of the subject, or by other parameters well known in the art (e.g., parameters specific to a specific tumor or viral infection). A "prophylactic" treatment is a treatment administered to a subject who does not exhibit signs of the disease or only exhibits early signs for the purpose of reducing the risk or severity of the developing condition.

[0233] Tumor or cancer or neoplasm: An abnormal growth of cells, which may be benign or malignant, usually but not always causing clinical symptoms. "Neoplastic" cell growth refers to cell growth that is unresponsive to physiological signals such as growth factors and inhibitory factors.

[0234] A "tumor" is a collection of neoplastic cells. In most cases, a tumor refers to a collection of neoplastic cells that forms a solid mass. Such tumors may be referred to as solid tumors. In some cases, the neoplastic cells may not form a solid mass, as in some leukemias. In such cases, the collection of neoplastic cells may be referred to as a liquid cancer.

[0235] Cancer is a malignant growth of neoplastic cells, either solid or fluid. Characteristics of cancer that define it as malignant include metastasis, interference with the normal function of neighboring cells, abnormal levels of cytokines or other secretions, suppression or exacerbation of one or more inflammatory or immune responses, and invasion of surrounding or distant tissues or organs, such as lymph nodes.

[0236] Tumors that present no substantial adverse clinical symptoms and / or grow slowly are termed "benign."

[0237] "Malignant" means that it is causing or may cause significant clinical symptoms in the future. Tumors that invade surrounding tissues and / or metastasize and / or produce substantial clinical symptoms through the production and secretion of chemical mediators that affect nearby or distant body systems are called "malignant."

[0238] "Metastatic disease" refers to cancer cells that have left the original tumor site and migrated to other parts of the body, for example, via the bloodstream, through the lymphatic system, or through a body cavity such as the peritoneal or thoracic cavity.

[0239] The amount of tumor in an individual is the “tumor burden.” Tumor burden can be measured as the number, volume, or mass of tumors and is typically assessed by physical examination, radiographic imaging, or pathological examination.

[0240] An "established" or "existing" tumor is one that is present at the time therapy is initiated. Typically, an established tumor can be identified by diagnostic testing. In some embodiments, an established tumor can be palpated. In some embodiments, an established tumor is at least 500 mm in size. 3 , such as at least 600 mm 3 , at least 700 mm 3 or at least 800 mm 3 In other embodiments, the tumor is at least 1 cm long. As for solid tumors, established tumors typically have a newly established and robust blood supply and may have induced regulatory T cells (Tregs) and myeloid-derived suppressor cells (MDSCs).

[0241] Unit Dose: A discrete quantity of a pharmaceutical composition containing a predetermined amount of the active ingredient.

[0242] Vesicle: A fluid-filled sac. In some embodiments, a vesicle is a sac comprising an amphiphilic substance. In some embodiments, the vesicle is a nanoparticle-based vesicle, which refers to a vesicle having a size or dimension on the nanometer scale. In some embodiments, a polymersome is a vesicle formed from one or more polymers.

[0243] definition:

[0244] As used herein, the term "optional" or "optionally" means that the subsequently described event or circumstance may or may not occur, and that the description includes instances where the event or circumstance occurs and instances where it does not. For example, "optionally substituted alkyl" means an alkyl group that may be substituted or unsubstituted.

[0245] It will be understood that substituents and substitution patterns on the compounds of the present invention can be selected by one skilled in the art to yield chemically stable compounds that can be readily synthesized from readily available starting materials by techniques known in the art, as well as those described below. If a substituent is itself substituted with more than one group, it will be understood that these multiple groups may be located on the same carbon or on different carbons, as long as a stable structure results.

[0246] As used herein, the term "optionally substituted" refers to the replacement of one to six hydrogen radicals in a given structure with radicals of specified substituents, including but not limited to hydroxy, hydroxyalkyl, alkoxy, halogen, alkyl, nitro, silyl, acyl, acyloxy, aryl, cycloalkyl, heterocyclyl, amino, aminoalkyl, cyano, haloalkyl, haloalkoxy, -OCO-CH2-O-alkyl, -OP(O)(O-alkyl)2 or -CH2-OP(O)(O-alkyl)2. Preferably, "optionally substituted" refers to the replacement of one to four hydrogen radicals in a given structure with the substituents mentioned above. More preferably, one to three hydrogen radicals are replaced by substituents as mentioned above. It should be understood that the substituents may be further substituted.

[0247] As used herein, the term "alkyl" refers to a saturated aliphatic group, including but not limited to C1-C 10 Straight chain alkyl or C1-C 10 Branched alkyl. Preferably, an "alkyl" group refers to a C1-C6 straight chain alkyl or a C1-C6 branched chain alkyl. Most preferably, an "alkyl" group refers to a C1-C4 straight chain alkyl or a C1-C4 branched chain alkyl. Examples of "alkyl" include, but are not limited to, methyl, ethyl, 1-propyl, 2-propyl, n-butyl, sec-butyl, tert-butyl, 1-pentyl, 2-pentyl, 3-pentyl, neopentyl, 1-hexyl, 2-hexyl, 3-hexyl, 1-heptyl, 2-heptyl, 3-heptyl, 4-heptyl, 1-octyl, 2-octyl, 3-octyl or 4-octyl. An "alkyl" group can be optionally substituted.

[0248] The term "acyl" is art-recognized and refers to a group represented by the general formula hydrocarbylC(O)-, preferably alkylC(O)-.

[0249] The term "acylamino" is art-recognized and refers to an amino group substituted with an acyl group and may be represented, for example, by the formula hydrocarbyl C(O)NH-.

[0250] The term "acyloxy" is art-recognized and refers to a group represented by the general formula hydrocarbylC(O)O-, preferably alkylC(O)O-.

[0251] The term "alkoxy" refers to an alkyl group having an oxygen attached thereto. Representative alkoxy groups include methoxy, ethoxy, propoxy, tert-butoxy, and the like.

[0252] The term "alkoxyalkyl" refers to an alkyl group substituted with an alkoxy group and can be represented by the general formula alkyl-O-alkyl.

[0253] The term "alkyl" refers to a saturated aliphatic group, including straight chain alkyl, branched chain alkyl, cycloalkyl (alicyclic) groups, alkyl substituted cycloalkyl and cycloalkyl substituted alkyl. In preferred embodiments, a straight chain or branched chain alkyl has 30 or fewer carbon atoms in its backbone (e.g., C for a straight chain 1-30 , for the branched chain C 3-30 ), and more preferably 20 or fewer carbon atoms.

[0254] Furthermore, the term "alkyl" as used throughout the specification, examples, and claims is intended to include both unsubstituted and substituted alkyl groups, wherein the latter refers to alkyl moieties having substituents replacing a hydrogen on one or more carbons in the hydrocarbon backbone, including haloalkyl groups such as trifluoromethyl and 2,2,2-trifluoroethyl, and the like.

[0255] The term "C x-y ” or “C x -C y " when used in conjunction with a chemical moiety such as acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy is intended to include groups containing x to y carbons in the chain. C0 alkyl indicates hydrogen if the group is in a terminal position and a bond if internal. For example, C 1-6 Alkyl groups contain one to six carbon atoms in the chain.

[0256] As used herein, the term "alkylamino" refers to an amino group substituted with at least one alkyl group.

[0257] As used herein, the term "alkylthio" refers to a thiol group substituted with an alkyl group, and can be represented by the general formula alkylS-.

[0258] As used herein, the term "amide" refers to a group

[0259] ,

[0260] where R 22 and R 23 Each independently represents hydrogen or a hydrocarbon group, or R 22 and R 23 Together with the N atom to which they are attached, the heterocycle has 4 to 8 atoms in the ring structure.

[0261] The terms "amine" and "amino" are art-recognized and refer to both unsubstituted and substituted amines and salts thereof, for example, a moiety represented by the formula

[0262] ,

[0263] where R 22 、R 23 and R24 Each independently represents hydrogen or a hydrocarbon group, or R 22 and R 23 Together with the N atom to which they are attached, the heterocycle has 4 to 8 atoms in the ring structure.

[0264] As used herein, the term "aminoalkyl" refers to an alkyl group substituted with an amino group.

[0265] As used herein, the term "aralkyl" refers to an alkyl group substituted with an aryl group.

[0266] As used herein, the term "aryl" includes substituted or unsubstituted aromatic carbocyclic rings and heteroaryl groups. The term "aryl" is used interchangeably with the term "aromatic group" herein. Unless otherwise specifically stated in the specification, the aryl moiety is optionally substituted with one or more substituents, which are independently alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl, trifluoromethoxy, nitro, trimethylsilyl, -OR a ,—SR a 、—OC(O)—Ra、—N(R a )2,—C(O)R a 、—C(O)OR a 、—OC(O)N(R a )2, —C(O)N(R a )2,—N(R a )C(O)OR a 、—N(R a )C(O)R a 、—N(Ra)C(O)N(R a )2,—N(R a )C(NR a )N(R a )2,—N(R a )S(O) t R a (where t is 1 or 2), —S(O) t OR a (where t is 1 or 2), —S(O) t N(R a )2 (where t is 1 or 2) or PO3(Ra )2, where each R a The term "aryl" also includes polycyclic ring systems having two or more cyclic rings, wherein two or more carbon atoms are shared by two adjacent rings, wherein at least one of the rings is aromatic, for example, the other cyclic rings can be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl and / or heterocyclic radicals. Aryl groups include benzene, naphthalene, phenanthrene, phenol, aniline, etc.

[0267] The term "carbamate" is art-recognized and refers to a group

[0268] ,

[0269] where R 22 and R 23 independently represents hydrogen or a hydrocarbon group.

[0270] As used herein, the term "carbocyclylalkyl" refers to an alkyl group substituted with a carbocyclyl group.

[0271] The term "carbocycle" includes 5-7 membered monocyclic and 8-12 membered bicyclic rings. Each ring of a bicyclic carbocycle can be selected from saturated, unsaturated, and aromatic rings. Carbocycle includes bicyclic molecules in which one, two, or three or more atoms are shared between the two rings. The term "fused carbocycle" refers to a bicyclic carbocycle in which each ring shares two adjacent atoms with the other ring. Each ring of a fused carbocycle can be selected from saturated, unsaturated, and aromatic rings. For example, an aromatic ring (e.g., phenyl) can be fused to a saturated or unsaturated ring (e.g., cyclohexane, cyclopentane, or cyclohexene). Any combination of saturated, unsaturated, and aromatic bicyclic rings is included in the definition of carbocycle, as long as valence permits. Exemplary "carbocycles" include cyclopentane, cyclohexane, bicyclo[2.2.1]heptane, 1,5-cyclooctadiene, 1,2,3,4-tetrahydronaphthalene, bicyclo[4.2.0]oct-3-ene, naphthalene, and adamantaneamine. Exemplary fused carbocycles include decahydronaphthalene, naphthalene, 1,2,3,4-tetrahydronaphthalene, bicyclo[4.2.0]octane, 4,5,6,7-tetrahydro-1H-indene, and bicyclo[4.1.0]hept-3-ene. A "carbocycle" may be substituted at any position or positions capable of carrying a hydrogen atom.

[0272] As used herein, the term "carbocyclylalkyl" refers to an alkyl group substituted with a carbocyclyl group.

[0273] The term "carbonate" is art-recognized and refers to the group -OCO2-.

[0274] As used herein, the term "carboxyl" refers to a group represented by the formula -CO2H.

[0275] As used herein, the term "ester" refers to the group -C(O)OR 22 , where R 22 Represents a hydrocarbon group.

[0276] As used herein, the term "ether" refers to a hydrocarbon group attached to another hydrocarbon group via an oxygen. Thus, an ether substituent of a hydrocarbon group can be hydrocarbon-O-. Ethers can be symmetrical or asymmetrical. Examples of ethers include, but are not limited to, heterocycle-O-heterocycle and aryl-O-heterocycle. Ethers include "alkoxyalkyl" groups, which can be represented by the general formula alkyl-O-alkyl.

[0277]

[0046] The terms "halo" and "halogen" as used herein mean halogen and include chlorine, fluorine, bromine and iodine.

[0278] As used herein, the terms "hetaralkyl" and "heteroaralkyl" refer to an alkyl group substituted with a heteroaryl group.

[0279] The terms "heteroaryl" and "hetaryl" include substituted or unsubstituted aromatic monocyclic ring structures, preferably 5- to 7-membered rings, more preferably 5- to 6-membered rings, whose ring structures include at least one heteroatom, preferably one to four heteroatoms, more preferably one or two heteroatoms. The terms "heteroaryl" and "hetaryl" also include polycyclic ring systems having two or more cyclic rings, wherein two or more carbon atoms are shared by two adjacent rings, and wherein at least one of the rings is heteroaromatic, for example, the other cyclic rings can be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl and / or heterocyclyl. Heteroaryl includes, for example, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyridine, pyrazine, pyridazine and pyrimidine.

[0280] As used herein, the term "heteroatom" means an atom of any element other than carbon or hydrogen. Preferred heteroatoms are nitrogen, oxygen, and sulfur.

[0281] As used herein, the term "heterocyclylalkyl" refers to an alkyl group substituted with a heterocyclyl group.

[0282] The terms "heterocyclyl," "heterocycle," and "heterocyclic" refer to substituted or unsubstituted non-aromatic ring structures, preferably 3 to 10-membered rings, more preferably 3 to 7-membered rings, whose ring structures include at least one heteroatom, preferably one to four heteroatoms, more preferably one or two heteroatoms. The terms "heterocyclyl" and "heterocyclic" also include polycyclic ring systems having two or more cyclic rings, wherein two or more carbon atoms are shared by two adjacent rings, and wherein at least one of the rings is heterocyclic, for example, the other cyclic rings can be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclyl. Heterocyclyl groups include, for example, piperidine, piperazine, pyrrolidine, morpholine, lactones, lactams, and the like.

[0283] As used herein, term "alkyl" refers to a group that is bonded to a carbon atom without an =O or =S substituent, and typically has at least one carbon-hydrogen bond and a primarily carbon backbone, but may optionally include heteroatoms. Therefore, for the purposes of this application, groups such as methyl, ethoxyethyl, 2-pyridyl, and even trifluoromethyl are considered to be alkyls, but substituents such as acetyl (which has a =O substituent on the connected carbon) and ethoxy (which is connected via oxygen rather than carbon) are not alkyls. alkyl includes, but is not limited to, aryl, heteroaryl, carbocycle, heterocycle, alkyl, alkenyl, alkynyl, and combinations thereof.

[0284] As used herein, the term "hydroxyalkyl" refers to an alkyl group substituted with a hydroxy group.

[0285] The term "lower" when used in conjunction with a chemical moiety such as acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy is intended to include groups having ten or fewer, preferably six or fewer atoms in the substituent. For example, "lower alkyl" refers to an alkyl group containing ten or fewer, preferably six or fewer carbon atoms. In certain embodiments, an acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy substituent as defined herein is a lower acyl, lower acyloxy, lower alkyl, lower alkenyl, lower alkynyl, or lower alkoxy group, respectively, whether occurring alone or in combination with other substituents, such as in the description of hydroxyalkyl and aralkyl (in such cases, for example, atoms in the aryl group are not counted when counting the carbon atoms in the alkyl substituent).

[0286] The terms "polycyclic group," "polycycle," and "polycyclic" refer to two or more rings (e.g., cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclyl) wherein two or more atoms are common to two adjacent rings, e.g., the rings are "fused rings." Each ring in the polycycle may be substituted or unsubstituted. In certain embodiments, each ring in the polycycle contains 3 to 10 atoms, preferably 5 to 7 atoms, in the ring.

[0287] The term "sulfate" is art-recognized and refers to the group -OSO3H or a pharmaceutically acceptable salt thereof.

[0288] The term "sulfonamide" is art-recognized and refers to a group represented by the general formula

[0289] ,

[0290] where R 22 and R 23 independently represents hydrogen or a hydrocarbon group.

[0291] The term "sulfoxide" is art-recognized and refers to the group -S(O)-.

[0292] The term "sulfonate" is art-recognized and refers to the group SO3H or a pharmaceutically acceptable salt thereof.

[0293] The term "sulfone" is art-recognized and refers to the group -S(O)2-.

[0294] The term "substituted" refers to a moiety that replaces a hydrogen on one or more carbons in the main chain with a substituent. It will be understood that "substituted" or "substituted with..." includes implicit conditions, i.e., such substitutions meet the allowed valences of the substituted atom and the substituent, and that the substitutions result in stable compounds, e.g., compounds that do not spontaneously undergo transformations such as those that occur through rearrangement, cyclization, elimination, etc. As used herein, the term "substituted" is considered to include all allowed substituents of organic compounds. Broadly speaking, allowed substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic substituents of organic compounds. For appropriate organic compounds, allowed substituents may be one or more and the same or different. For purposes of the present invention, heteroatoms such as nitrogen may have hydrogen substituents as described herein and / or any allowed substituents of organic compounds that satisfy the valences of the heteroatoms. Substituents may include any substituent described herein, for example, halogen, hydroxy, carbonyl (e.g., carboxyl, alkoxycarbonyl, formyl, or acyl), thiocarbonyl (e.g., thioester, thioacetate, or thioformate), alkoxy, phosphoryl, phosphate, phosphonate, phosphinate, amino, amido, amidine, imine, cyano, nitro, azido, sulfhydryl, alkylthio, sulfate, sulfonate, sulfamoyl, sulfonamido, sulfonyl, heterocyclyl, aralkyl, or aromatic or heteroaromatic moieties. It will be understood by those skilled in the art that the moieties substituted on the hydrocarbon chain may themselves be substituted, if appropriate.

[0295] As used herein, the term "thioalkyl" refers to an alkyl group substituted with a thiol group.

[0296] As used herein, the term "thioester" refers to the group -C(O)SR 22 or –SC(O)R 22 , where R22 Represents a hydrocarbon group.

[0297] As used herein, the term "thioether" is equivalent to an ether in which the oxygen is replaced by sulfur.

[0298] The term "urea" is art-recognized and may be represented by the general formula

[0299] ,

[0300] where R 22 and R 23 independently represents hydrogen or a hydrocarbon group.

[0301] The term "aromatic amino acid" includes amino acids with side chains containing aromatic groups, such as phenylalanine, tyrosine, or tryptophan. An aromatic group refers to the portion of a molecule that contains an aromatic ring. For example, phenylalanine is an aromatic amino acid that contains an aromatic group (i.e., a benzyl group). Phenylalanine (Phe) and tryptophan (Trp) are prototypical aromatic amino acids.

[0302] Those of ordinary skill in the art will appreciate that the definitions provided above are not intended to include substitution patterns that are not permitted (e.g., a methyl group substituted with 5 different groups, etc.). Those of ordinary skill in the art can easily identify such substitution patterns that are not permitted. Any functional group disclosed herein and / or defined above may be substituted or unsubstituted, unless otherwise indicated herein. Unless otherwise explained, all technical and scientific terms used herein have the same meaning as those generally understood by those skilled in the art to which the present disclosure pertains. The term "comprising" means "including." Thus, comprising "A" or "B" means including A, including B, or including both A and B. It should be further understood that all base sizes or amino acid sizes and all molecular weight or molecular mass values ​​given for nucleic acids or polypeptides are approximate and are provided for description. Although methods and materials similar or equivalent to the methods and materials described herein can be used in the practice or testing of the present disclosure, suitable methods and materials are described herein. In the event of a conflict, this specification (including term explanation) shall prevail. In addition, materials, methods, and examples are illustrative only and are not intended to be limiting.

[0303] Description of the implementation plan

[0304] Provided herein are compositions of particles comprising an amphiphile and a drug molecule that can be used for the treatment or prevention of a disease, such as one or more cancers, one or more autoimmune diseases, one or more allergies, and / or one or more infectious diseases. Particles comprising certain compositions of an amphiphile and a peptide antigen conjugate have particular utility for use as vaccines for the treatment or prevention of a disease, such as the prevention or treatment of one or more cancers, one or more autoimmune diseases, one or more allergies, and / or one or more infectious diseases.

[0305] The present disclosure relates to a vaccine comprising at least one peptide antigen conjugate having a formula selected from the group consisting of PEG-[E1]-A-[E2]-[U]-H and H-[U]-[E1]-A-[E2]-PEG

[0306] in

[0307] A is a peptide antigen;

[0308] E1 is the N-terminal extension;

[0309] E2 is the C-terminal extension;

[0310] H is independently a hydrophobic block at each occurrence, wherein one or more drug molecules (D) are optionally attached to each H directly or via a suitable linker X1;

[0311] U is independently a linker at each occurrence;

[0312] [ ] indicates that the group is optional, and

[0313] - means that two adjacent groups are directly attached to each other by a covalent bond or indirectly attached to each other via a suitable linker X.

[0314] In vaccine embodiments wherein (i) the average aqueous solubility of the peptide antigen (A) of one or more peptide antigen conjugates is less than 1 mg / mL, or (ii) the average GRAVY score of the peptide antigen (A) of one or more peptide antigen conjugates is > 0, an amphiphile is present.

[0315] The present disclosure relates to a vaccine comprising at least one peptide antigen conjugate having a formula selected from the group consisting of PEG-[E1]-A-[E2]-[U]-H and H-[U]-[E1]-A-[E2]-PEG, and an amphiphile having the formula S-[B]-[U]-H,

[0316] in

[0317] A is a peptide antigen;

[0318] E1 is the N-terminal extension;

[0319] E2 is the C-terminal extension;

[0320] H is independently a hydrophobic block at each occurrence, wherein one or more drug molecules (D) are optionally attached to each H directly or via a suitable linker X1;

[0321] S is the solubilization block;

[0322] B is a spacer;

[0323] U is independently a linker at each occurrence;

[0324] [ ] indicates that the group is optional, and

[0325] - means that two adjacent groups are directly attached to each other by a covalent bond or indirectly attached to each other via a suitable linker X.

[0326] In one embodiment of the vaccine, the PEG group of the peptide antigen conjugate includes a terminal functional group selected from OH, MeO- and NH2.

[0327] In a specific embodiment of the vaccine, the PEG group of the peptide antigen conjugate is polyethylene glycol.

[0328] In some embodiments of the vaccine, the PEG group of the peptide antigen conjugate comprises between 4 and 36 monomer units.

[0329] In specific embodiments of the vaccine, the PEG group of the peptide antigen conjugate comprises between 4 and 12 monomeric units, or between 12 and 36 monomeric units, preferably 24 monomeric units.

[0330] In some embodiments of the vaccine, where an amphiphile is present, the amphiphile comprises a dendrimer amplifier.

[0331] In some embodiments of the vaccine, the amphiphile S comprises a dendrimer amplifier. In other embodiments, the amphiphile S has a dendritic architecture.

[0332] In some embodiments of the vaccine, S of the amphiphile comprises two or more solubilizing groups (SG). In other embodiments, two or more SGs are linked to the rest of S via a dendrimer amplifier, for example, 4 to 8 SGs are linked to S.

[0333] In some embodiments of the vaccine, SG is independently selected from amines, hydroxyls, carboxylic acids and / or sugar molecules, wherein the sugar molecules are independently selected from mannose, glucose, glucosamine, N-acetylglucose, galactose, galactosamine, N-acetylgalactosamine, N-acetylglucosamine, phosphoserine and any derivatives thereof, agonists of CD22a, sialyl Lewis x, and combinations thereof.

[0334] In some embodiments of the vaccine, the dendrimer amplifier comprises 1 to 10 generations of repeating monomer units, each generation having between 2 and 6 branches. In other embodiments, the dendrimer amplifier comprises 2 to 3 generations of repeating monomer units, each generation having between 2 and 3 branches. In some embodiments of the vaccine, the repeating monomer units are selected from FG1-(CH2) y2 CH(R 1 )2、FG1-(CH2) y2 C(R 1 )3、FG1-(CH2CH2O) y2 CH(R 1 )2、FG1-(CH2CH2O) y2 C(R 1 )3、and FG1-CH(R 1 )2、FG1-C(R 1 )3, wherein R1 at each occurrence is independently selected from (CH2) y3 -FG2, (OCH2CH2) y3 -FG2 and CH2(OCH2CH2) y3 -FG2); y2 and y3 are each independently an integer of 1 to 6 repeating units at each occurrence; FG1 is a first functional group; and FG2 is a second functional group. In some embodiments, FG1 is -NH2; and FG2 is independently -CO2- or -CO2H at each occurrence. In some embodiments, FG1 is independently -CO2- or -CO2H at each occurrence; and FG2 is -NH2.

[0335] In some embodiments of the vaccine, SG is linked to S via a suitable linker X5. In some embodiments of the vaccine, the suitable linker X5 linking SG to S is selected from a lower alkyl group and a PEG group. In some embodiments of the vaccine, two or more SG are linked to the rest of S via a dendrimer amplifier via a suitable linker X5, which links the two or more SG to the terminal functional group (FGt) of the dendrimer amplifier via an amide bond. In some embodiments, the linker X5 linking SG to the dendrimer amplifier is selected from -NH-R 19 、-NH-C(O)-R 19 、-C(O)-NH-R 19 -or-C(O)-R 19 , where R 19 Can be selected from but not limited to -(CH2) t -、-(CH2CH2O) t -CH2CH2-, -(CH2)tC(O)-NH-(CH2) u-、-(CH2CH2O) t CH2CH2C(O)-NH-(CH2) u -、-(CH2) t -NH-C(O)-NH-(CH2) u -or-(CH2CH2O) t CH2CH2NH-C(O)-(CH2) u -, wherein t and u are each independently an integer generally selected from between 1 and 6, such as 1, 2, 3, 4, 5 or 6.

[0336] In some embodiments of the vaccine, the dendrimer amplifier comprises a polyethylene oxide (PEG) group.

[0337] In some embodiments of the vaccine, H of the amphiphile comprises a higher alkane, an aromatic group, a fatty acid, a sterol, a polyunsaturated hydrocarbon, squalene, a saponin, and / or a polymer.

[0338] In some embodiments of the vaccine, H of the peptide antigen conjugate comprises a higher alkane, an aromatic group, a fatty acid, a sterol, a polyunsaturated hydrocarbon, and / or a polymer.

[0339] In some embodiments of the vaccine, each H independently comprises a poly(amino acid) comprising a monomer selected from the group consisting of a hydrophobic amino acid (M), a reactive amino acid (N), a spacer amino acid (O), a charged amino acid (P), and combinations thereof, with the proviso that at least one of M or N is present.

[0340] In some embodiments of the vaccine, each H independently comprises a poly(amino acid) having the formula:

[0341] ,

[0342] wherein M, N, O and P are each independently present or absent, provided that at least one of M or N is present;

[0343] m, n, o and p each independently represent an integer from 1 to 100, and the sum of m, n, o and p is less than or equal to 100;

[0344] R 3 Selected from hydrogen, NH2, NH-CH3, NH-(CH2) y5 CH3, OH or a drug molecule (D) directly or via a suitable linker X1; and

[0345] y5 is an integer selected from 1 to 6.

[0346] In some embodiments of the vaccine, P is absent. In other embodiments, N, O, and P are each absent.

[0347] In some embodiments of the vaccine, P is , where each R 5 Independently, a group comprising 1 to 2 charged functional groups.

[0348] In some embodiments of the vaccine, O is , wherein each Q is independently selected from (CH2) y6 and (CH2CH2O) y7 CH2CH2; each y6 is independently selected from an integer from 1 to 6; and each y7 is independently selected from an integer from 1 to 4.

[0349] In some embodiments of the vaccine, N is , wherein each X1 is independently a suitable linker; and each D is independently a drug molecule. In some embodiments of the vaccine, X1 is absent. In other embodiments, X1 is present and is selected from a lower alkyl group and a PEG group. In other embodiments, X1 is present and is selected from an enzyme-cleavable linker and a pH-sensitive linker. In some embodiments of the vaccine, X1 is present and comprises an enzyme-degradable peptide and / or a self-immolative linker.

[0350] In some embodiments, X1 is present and is selected from -(CH2) y10 -W and -(CH2) y10 -R 6 , wherein y10 is an integer selected from 1 to 6, and R 6 Any one or more selected from the following: -C(O)-NH-R 7 、-NH-C(O)-R 7 、-NH-C(O)-OR 7 、-OC(O)-NH-R 7 、-OC(O)-R 7 、-C(O)-OR 7 、-OR 7 , OC(O)-W or -C(O)-W, where R 7 Any one or more selected from the following: -(CH2) y11 -W, -(CH2) y11 -(OCH2CH2) y12 -W, -(CH2) y11 -(OCH2CH2) y12 -(CH2) y13 -W, –CHR 8 -C(O)-W, –CHR 8-C(O)-(NH-CHR 8 -C(O)) j -W、–(CH2) y11 -C(O)-NH-CHR 8 -C(O)-W、–(CH2) y11 -C(O)-NH-CHR 8 -C(O)-(NH-CHR 8 -C(O)) j -W、–(CH2) y11 -(OCH2CH2) y12 -C(O)-NH-CHR 8 -C(O)-W、–(CH2) y11 -(OCH2CH2) y12 -(CH2) y13 C(O)-NH-CHR 8 -C(O)-W、–(CH2) y11 -(OCH2CH2) y12 -C(O)-NH-CHR 8 -C(O)-(NH-CHR 8 -C(O)) j -W、–(CH2) y11 -(OCH2CH2) y12 -(CH2) y13 -C(O)-NH-CHR 8 -C(O)-(NH-CHR 8 -C(O)) j -W、–CHR 8 -C(O)-NH-C6H4-CH2-O-C(O)-W、–CHR 8 -C(O)-NH(CH3)(CH2)2-O-C(O)-W、–CHR 8 -C(O)-(NH-CHR 8 -C(O)) j -NH-C6H4-CH2-O-C(O)-W、–CHR 8 -C(O)-(NH-CHR 8 -C(O)) j -NH(CH3)(CH2)2-O-C(O)-W、–(CH2) y11 -C(O)-(NH-CHR 8 -C(O)) j -NH-C6H4-CH2-O-C(O)-W、–(CH2) y11 -C(O)-(NH-CHR 8-C(O)) j -NH(CH3)(CH2)2-O-C(O)-W、–(CH2) y11 -(OCH2CH2) y12 -C(O)-(NH-CHR 8 -C(O)) j -NH-C6H4-CH2-O-C(O)-W、–(CH2) y11 -(OCH2CH2) y12 -C(O)-(NH-CHR 8 -C(O)) j -NH(CH3)(CH2)2-O-C(O)-W、–(CH2) y11 -(OCH2CH2) y12 -(CH2) y13 C(O)-(NH-CHR 8 -C(O)) j -NH-C6H4-CH2-O-C(O)-W、–(CH2) y11 -(OCH2CH2) y12 -(CH2) y13 C(O)-(NH-CHR 8 -C(O)) j -NH(CH3)(CH2)2-O-C(O)-W、–(CH2) y11 -(OCH2CH2) y12 -(CH2) y13 -C(O)-NH-(CH2) y14 -C(O)-(NH-CHR 8 -C(O)) j -NH-C6H4-CH2-O-C(O)-W、–(CH2) y11 -(OCH2CH2) y12 -(CH2) y13 C(O)-NH-(CH2) y14 -C(O)-(NH-CHR 8 -C(O)) j -NH(CH3)(CH2)2-O-C(O)-W、–(CH2) y11 -(OCH2CH2) y12 -C(O)-NH-(CH2) y14 -C(O)-(NH-CHR 8 -C(O)) j -NH-C6H4-CH2-O-C(O)-W、–(CH2) y11 -(OCH2CH2)y12 -C(O)-NH-(CH2) y14 -C(O)-(NH-CHR 8 -C(O)) j -NH(CH3)(CH2)2-OC(O)-W、-CHR 8 -C(O)-NH-(CH2) y15 -W, -CHR 8 -NH-C(O)-(CH2) y15 -W, -CHR 8 -C(O)-(NH-CHR 8 -C(O)) j -NH-(CH2) y15 -W, -CHR 8 -NH-(C(O)-CHR 8 -NH) j -C(O)-(CH2) y15 -W, wherein y11, y12, y13, y14, y15 and j are each independently selected from an integer selected from 1 to 6, R 8 is any amino acid side group, and W can be independently selected from H (hydrogen), FG3, LG and w; wherein FG3 is any suitable functional group for attachment to a functional group ("FG4") present on the drug molecule, said FG3 can be selected from, but not limited to, carboxylic acids, activated carboxylic acids (e.g., carbonyl thiazolidine-2-thione ("TT"), NHS or nitrophenol esters), carboxylic anhydrides, amines and protected amines (e.g., tert-butoxycarbonyl protected amines), OSi(CH3), alkenes, azides, alkynes, strained alkynes, halogens (e.g., fluorine, chlorine), alkenes and endocyclic alkenes (e.g., allyl), CN, OH and epoxy groups, hydrazines (including hydrazides), carbohydrazides, aldehydes, ketones, carbamates and activated carbamate, LG is any suitable leaving group, which can be selected from any suitable leaving group (e.g., NHS, TT, nitrophenol, etc.); and w is a group resulting from the reaction of FG4 with FG3 or the replacement of LG with FG4, and is typically selected from NH-, C(O)-, NH-C(O)-, C(O)-NH-, OC(O)-NH-, C(O)-NH-N=C(CH3)-, NH-N=C(CH3)- or -C(CH3)=N-NH-C(O)-, wherein w is always directly connected to D (i.e., wD) or indirectly connected to D via X3 (i.e., w-X3-D).

[0351] In some embodiments of the vaccine, M is , where each R 4 are independently hydrophobic groups.

[0352] In some embodiments of the vaccine, R4 yes

[0353]

[0354] in

[0355] α is aryl or heteroaryl;

[0356] X2 is present or absent and, when present, is a suitable linker;

[0357] y8 is an integer selected from 0 and 6; and

[0358] Z 1 、Z 2 and Z 3 Each is independently selected from H, F, hydroxy, amino, alkyl and fluoroalkyl.

[0359] In some embodiments of the vaccine, α is an aryl group, for example, phenyl or naphthyl. In other embodiments, α is a heteroaryl group, for example, pyridyl, quinolyl, isoquinolyl, indolyl, or benzimidazolyl.

[0360] In some embodiments of the vaccine, X2 is absent. In other embodiments, X2 is present and is selected from C(O), CO2(CH2) y9 、CO2、C(O)NH(CH2) y9 , NHC(O) and NHC(O)(CH2) y9 , wherein y9 is an integer generally selected from 1 to 6. In other embodiments, X2 is present and is selected from lower alkyl and PEG groups.

[0361] In some embodiments of the vaccine, each R 4 Independently selected from:

[0362] 、 、 、

[0363] 、 、 、

[0364] 、 、 、

[0365] 、 、 、

[0366] and ,

[0367] wherein each X2 is independently selected from a suitable linker, and each y8 is independently selected from an integer between 0 and 6. In other embodiments, each R 4 Independently selected from:

[0368] 、 、 、

[0369] 、 、 、

[0370] 、 、 、

[0371] 、 、 、

[0372] and ,

[0373] wherein each y8 is independently selected from an integer between 0 and 6. In other embodiments, each R 4 Independently selected from:

[0374] 、 、 、 、 、 、 、 、 、 、 、 and In other embodiments, each R 4 Independently selected from:

[0375] 、 、 、

[0376] 、 、

[0377] 、 、 、

[0378] 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 and In a preferred embodiment, each R 4 Independently selected from:

[0379] 、 、 、 and .

[0380] In some embodiments of the vaccine, wherein at least one D is:

[0381]

[0382] in,

[0383] R 20 is selected from H, alkyl, alkoxyalkyl, aryl, heteroaryl, aminoalkyl, amide, and ester; and X 3 is selected from the group consisting of alkyl, alkoxyalkyl, aralkyl, heteroaralkyl, aryl, heteroaryl and carboxyl.

[0384] In some embodiments of the vaccine, R 20 is selected from H, alkyl and alkoxyalkyl; and X 3 In other embodiments, R 20 It's butyl.

[0385] In some embodiments of the vaccine, X 3 It is an alkyl group.

[0386] In some embodiments of the vaccine, m, n, o, and p each independently represent an integer from 1 to 30, and the sum of m, n, o, and p is less than or equal to 30.

[0387] In some embodiments of the vaccine, m, n, o, and p each independently represent an integer from 1 to 10, and the sum of m, n, o, and p is less than or equal to 10.

[0388] In some embodiments of the vaccine, B is present and is a hydrophilic polymer, e.g., a PEG group. In other embodiments, B is present and is a hydrophilic peptide.

[0389] In some embodiments of the vaccine, the PEG group comprises between 4 and 36 monomeric units. In other embodiments, the PEG group comprises between 4 and 12 monomeric units.

[0390] In some embodiments of the vaccine, the hydrophilic peptide comprises between 4 and 36 amino acids. In other embodiments, the hydrophilic peptide comprises between 4 and 12 amino acids.

[0391] In some embodiments of the vaccine, the amphiphile has the formula SH. In other embodiments, the amphiphile has the formula SBUH. In other embodiments, the amphiphile has the formula SBUHD.

[0392] In some embodiments of the vaccine, the vaccine comprises a molar ratio of peptide antigen conjugate to amphiphile between about 4:1 and about 1:20, preferably about 1:1.

[0393] In some embodiments of the vaccine, the vaccine is a cancer vaccine, an infectious disease vaccine, a tolerance-inducing allergy vaccine, a tolerance-inducing autoimmune disease vaccine, or a tolerance-inducing transplant rejection vaccine.

[0394] In some embodiments of the vaccine, the peptide antigen (A) comprises a sequence in which one or more cysteine ​​residues have been replaced by α-aminobutyric acid and / or one or more methionine residues have been replaced by norleucine. In some embodiments of the vaccine, at least one peptide antigen conjugate comprises A selected from minimal immunogens. Minimal immunogens are, for example, small peptide fragments derived from naturally occurring proteins that contain B cell epitopes. Minimal immunogens can be used in cancer vaccines, infectious disease vaccines, and vaccines that induce tolerance, as well as in the treatment of cardiovascular disease or neurodegenerative diseases.

[0395] In some embodiments of the vaccine, A is a peptide antigen selected from the group consisting of RGYLTKILHVFHGLLPGFLVKMSGDLLE (SEQ ID NO:52), PGFLVKMSGDLLE (SEQ ID NO:53), PGFLVKnSGDLLE, wherein n = norleucine (SEQ ID NO:54); PGFLVKMSSDLLG (SEQ ID NO:55), PGFLVKnSSDLLG, wherein n is norleucine (SEQ ID NO:56); SIPWNLERITPPR (SEQ ID NO:57); SIPWNLERITPPR (SEQ ID NO:58); SIPWNLE (SEQ ID NO:59); SIPWNLEKVTPPR (SEQ ID NO:60); SIPWNLDRVTPPR (SEQ ID NO:61); NVPEEDGTRFHRQASKC (SEQ ID NO:62); NVPEEDGTRFHRQASK (SEQ ID NO:63); PEEDGTR (SEQ ID NO:64). NO:64), NVPEEDG (SEQ ID NO:65); NVPEEDATRFHRQGSK (SEQ ID NO:66); LFAPGEDIIGASSDCSTCFVSQSGTSQAAA (SEQ ID NO:67);CSTCFVSQSGTSQAAA (SEQ ID NO:68); STCFVSQSGTSQAAA (SEQ ID NO:69), STBFVSQSGTSQAAA (SEQ ID NO:70); STBFVSQ (SEQ ID NO:71); NO:72); EPKSRFAMLDDVKILANGLLQLGHGLKDFVHKTKGQIND (SEQ ID NO:73); NO:77); LKDFVHKTKGQIND (SEQ ID NO:78); RFAMLDDVKILANGLLQLGH (SEQ ID NO:79); GLLQLGHGLKDFVHKTKGQI (SEQ ID NO:80); and IFQKLNIFDQSFYDLSLQTSEIKEEEKELRRTTYKLQVKNEEVKNMSLELNSKLESLLEEKILLQQKVK (SEQ ID NO:81). ;

[0396] In some embodiments of the vaccine, A is directly attached to El by a covalent bond, and El is directly attached to H or indirectly attached to H via U by a covalent bond.

[0397] In some embodiments of the vaccine, A is directly attached to E2 via a covalent bond, and E2 is directly attached to H or indirectly attached to H via U via a covalent bond.

[0398] In some embodiments of the vaccine, E1 and E2 each comprise a PEG group of between 4 and 36 monomeric units, eg, the PEG group comprises between 4 and 24 monomeric units.

[0399] In some embodiments of the vaccine, E1 and E2 each comprise a peptide.

[0400] In some embodiments of the vaccine, the peptide comprises 4 to 24 amino acids.

[0401] In some embodiments of the vaccine, at least one peptide antigen conjugate comprises A selected from the group consisting of an autoantigen, an alloantigen, and an allergen.

[0402] In some embodiments of the vaccine, the S of the amphiphile comprises two or more solubilizing groups (SG) independently selected from carboxylic acids, phosphoserines and / or sugar molecules, wherein the sugar molecules are independently selected from mannose, glucose, glucosamine, N-acetylglucose, galactose, galactosamine, and N-acetylgalactosamine and agonists of CD22a.

[0403] In some embodiments of the vaccine, the vaccine comprises at least one D selected from the group consisting of an inhibitor of mTOR, RORγt, CDK8 / 19, and HDAC, and an inhibitor of AHR, RAR, and A 2a In some embodiments of the vaccine, the at least one D is selected from ATP-competitive mTOR inhibitors.

[0404] In some embodiments of the vaccine, the vaccine further comprises a second drug molecule (D2) independently selected from the group consisting of inhibitors of mTOR, RORγt, CDK8 / 19, and HDAC; inhibitors of AHR, RAR, and A 2a and an immunostimulatory agent selected from the group consisting of an agonist of NLR, CLR, TLR, and STING, with the proviso that D and D2 bind to different receptors.

[0405] In some embodiments of the vaccine, the at least one D is selected from an inhibitor of mTOR and an agonist of AHR, and the D2 is selected from an agonist of NLR, CLR, TLR and STING. In some embodiments of the vaccine, the at least one D is selected from an ATP-competitive mTOR inhibitor, and the D2 is selected from an agonist of NLR, CLR, TLR and STING.

[0406] In some embodiments of the vaccine, the D2 is selected from agonists of TLR-3, TLR-7, TLR-8, TLR-7 / 8, TLR-9, and STING. In some embodiments of the vaccine, the D2 is selected from RNA and imidazoquinoline agonists of TLR-7, TLR-8, and TLR-7 / 8.

[0407] In some embodiments of the vaccine, the vaccine further comprises a third drug molecule (D3) independently selected from the group consisting of inhibitors of mTOR, RORγt, CDK8 / 19 and HDAC; inhibitors of AHR, RAR and A 2a and an immunostimulatory agent selected from the group consisting of an agonist of NLR, CLR, TLR and STING, with the proviso that D, D2 and D3 bind to different receptors.

[0408] In some embodiments of the vaccine, the at least one D is selected from AZD-8055, AZD-2016, KU-0063794, CC223, Torin-1, Torin-2, INK-128, WYE354, WYE132, OSI-027, OXA-01, PI-103, NVP-BEZ235, GNE-493, GSK2126458, rapamycin, tacrolimus, everolimus, RAD001, CCI-779, and AP23573.

[0409] In some embodiments of the vaccine, the molar ratio of total peptide antigen conjugate to at least one D is between about 20:1 and 1:2, or between about 10:1 and about 1:1, or between about 4:1 and about 2:1, preferably about 1:1.

[0410] In some embodiments of the vaccine, at least one peptide antigen conjugate comprises A selected from a tumor antigen.

[0411] In some embodiments of the vaccine, the S of the amphiphile comprises two or more solubilizing groups (SG) independently selected from amines or sugar molecules, wherein the sugar molecules are independently selected from mannose and sialyl Lewis x, and combinations thereof. In some embodiments of the vaccine, the S of the amphiphile comprises two or more solubilizing groups (SG) independently selected from amines, carboxylic acids, or sugar molecules, wherein the sugar molecules are independently selected from mannose, sialyl Lewis x, sialyl Lewis a, Lewis y, Lewis x, Tn, sTn, TF, sTF, Globo H, SSEA-3, GM2, GD2, GD3, and fucosyl GM1, and combinations thereof.

[0412] In some embodiments of the vaccine, each H of the amphiphile and / or peptide antigen conjugate independently comprises a poly(amino acid) containing a monomer comprising a hydrophobic amino acid (M) selected from the following: tryptophan, 1-methyltryptophan and p-aminophenylalanine. In other embodiments, each H of the amphiphile and / or peptide antigen conjugate comprises a poly(amino acid) containing a monomer comprising a reactive amino acid (N), wherein the monomer comprises a D selected from Glu-TLR-7 / 8a. In some embodiments of the vaccine, at least one D is present and selected from an agonist of TLR-3, TLR-7, TLR-8, TLR-7 / 8, TLR-9 and STING. In some embodiments of the vaccine, the vaccine further comprises a second drug molecule (D2) selected from an inhibitor of mTOR. In other embodiments, D2 is selected from rapamycin, tacrolimus, everolimus, RAD001, CCI-779 and AP23573. In some embodiments of the vaccine, the molar ratio of the peptide antigen conjugate to D2 is between about 20:1 and 1:2, or between about 10:1 and about 1:1, or between about 4:1 and about 2:1, preferably about 1:1.

[0413] In some embodiments of the vaccine, A is a glycopeptide. In other embodiments, A is selected from HGVT*S*APDT*RPAPGS*T*APPA (SEQ ID NO: 534), DT*RPAPGS*T*APPAHGVT*S*AP (SEQ ID NO: 535), GS*T*APPAHGVT*S*APDT*RPAPGS*T*APPA (SEQ ID NO: 536), GVT*S*APDT*RPAP (SEQ ID NO: 537), APDT*RPAPGS*T*A (SEQ ID NO: 538), GS*T*APPAHGVT*S*AP (SEQ ID NO: 539), VT*S*AP (SEQ ID NO: 540), DT*RPAP (SEQ ID NO: 541), and GS*T*AP (SEQ ID NO: 542). 542), wherein * is an O-linked glycan and is independently selected at each occurrence from sialyl Lewis x, sialyl Lewis a, Lewis y, Lewis x, Tn, sTn, TF, sTF.

[0414] In some embodiments of the vaccine, S of the amphiphile comprises a second or third generation dendrimer; and B comprises 4 to 36 PEG monomer units.

[0415] In some embodiments of the vaccine, S of the amphiphile comprises a second or third generation dendrimer; and H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0416] In some embodiments of the vaccine, B comprises 4 to 36 monomer units; and H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0417] In some embodiments of the vaccine, S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; and H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0418] In some embodiments of the vaccine, S of the amphiphile comprises a second or third generation dendrimer; and H of the amphiphile comprises a poly(amino acid) comprising p-aminophenylalanine.

[0419] In some embodiments of the vaccine, B comprises 4 to 36 PEG monomer units; and H of the amphiphile comprises a polymer of p-aminophenylalanine.

[0420] In some embodiments of the vaccine, S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; and H of the amphiphile comprises a polymer of p-aminophenylalanine.

[0421] In some embodiments of the vaccine, S of the amphiphile comprises a second or third generation dendrimer; and H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and an imidazoquinoline-containing reactive amino acid (N).

[0422] In some embodiments of the vaccine, B comprises 4 to 36 PEG monomer units; and H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and an imidazoquinoline-containing reactive amino acid (N).

[0423] In some embodiments of the vaccine, S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; and H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and an imidazoquinoline-containing reactive amino acid (N).

[0424] In some embodiments of the vaccine, S of the amphiphile comprises a second or third generation dendrimer; and H of the amphiphile comprises a poly(amino acid) comprising tryptophan and an imidazoquinoline-reactive amino acid (N).

[0425] In some embodiments of the vaccine, B comprises 4 to 36 PEG monomer units; and H of the amphiphile comprises a poly(amino acid) comprising tryptophan and an imidazoquinoline-containing reactive amino acid (N).

[0426] In some embodiments of the vaccine, S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; and H of the amphiphile comprises a poly(amino acid) of tryptophan and an imidazoquinoline-containing reactive amino acid (N).

[0427] In some embodiments of the vaccine, S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; and SG comprises mannose.

[0428] In some embodiments of the vaccine, S of the amphiphile comprises a second or third generation dendrimer; H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M); and SG comprises mannose.

[0429] In some embodiments of the vaccine, B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M); and SG comprises mannose.

[0430] In some embodiments of the vaccine, S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M); and SG comprises mannose.

[0431] In some embodiments of the vaccine, S of the amphiphile comprises a second or third generation dendrimer; H of the amphiphile comprises a polymer of p-aminophenylalanine; and SG comprises mannose.

[0432] In some embodiments of the vaccine, B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a polymer of p-aminophenylalanine; and SG comprises mannose.

[0433] In some embodiments of the vaccine, S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a polymer of p-aminophenylalanine; and SG comprises mannose.

[0434] In some embodiments of the vaccine, S of the amphiphile comprises a second or third generation dendrimer; and H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and an imidazoquinoline-containing reactive amino acid (N); and SG comprises mannose.

[0435] In some embodiments of the vaccine, B comprises 4 to 36 PEG monomer units; and H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and an imidazoquinoline-containing reactive amino acid (N); and SG comprises mannose.

[0436] In some embodiments of the vaccine, S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and an imidazoquinoline-containing reactive amino acid (N); and SG comprises mannose.

[0437] In some embodiments of the vaccine, S of the amphiphile comprises a second or third generation dendrimer; and H of the amphiphile comprises a poly(amino acid) of tryptophan and an imidazoquinoline-containing reactive amino acid (N); and SG comprises mannose.

[0438] In some embodiments of the vaccine, B comprises 4 to 36 PEG monomer units; and H of the amphiphile comprises a poly(amino acid) of tryptophan and an imidazoquinoline-containing reactive amino acid (N); and SG comprises mannose.

[0439] In some embodiments of the vaccine, S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; and H of the amphiphile comprises a poly(amino acid) of tryptophan and an imidazoquinoline-reactive amino acid (N); and SG comprises mannose.

[0440] In some embodiments of the vaccine, S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; and H of the peptide antigen conjugate comprises a poly(amino acid) containing a hydrophobic amino acid (M).

[0441] In some embodiments of the vaccine, S of the amphiphile comprises a second or third generation dendrimer; H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M); and H of the peptide antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0442] In some embodiments of the vaccine, B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M); and H of the peptide antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0443] In some embodiments of the vaccine, S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M); and H of the peptide antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0444] In some embodiments of the vaccine, S of the amphiphile comprises a second or third generation dendrimer; H of the amphiphile comprises a polymer of p-aminophenylalanine; and H of the peptide antigen conjugate comprises a poly(amino acid) containing a hydrophobic amino acid (M).

[0445] In some embodiments of the vaccine, B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a polymer of p-aminophenylalanine; and H of the peptide antigen conjugate comprises a poly(amino acid) containing a hydrophobic amino acid (M).

[0446] In some embodiments of the vaccine, S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a polymer of p-aminophenylalanine; and H of the peptide antigen conjugate comprises a poly(amino acid) containing a hydrophobic amino acid (M).

[0447] In some embodiments of the vaccine, S of the amphiphile comprises a second or third generation dendrimer; H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and an imidazoquinoline-containing reactive amino acid (N); and H of the peptide antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0448] In some embodiments of the vaccine, B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and a reactive amino acid (N) containing an imidazoquinoline; and H of the peptide antigen conjugate comprises a poly(amino acid) containing a hydrophobic amino acid (M).

[0449] In some embodiments of the vaccine, S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and an imidazoquinoline-containing reactive amino acid (N); and H of the peptide antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0450] In some embodiments of the vaccine, S of the amphiphile comprises a second or third generation dendrimer; H of the amphiphile comprises a poly(amino acid) comprising tryptophan and an imidazoquinoline-reactive amino acid (N); and H of the peptide antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0451] In some embodiments of the vaccine, B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a poly(amino acid) comprising tryptophan and an imidazoquinoline-reactive amino acid (N); and H of the peptide antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0452] In some embodiments of the vaccine, S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a poly(amino acid) comprising tryptophan and an imidazoquinoline-reactive amino acid (N); and H of the peptide antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0453] In some embodiments of the vaccine, S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; SG comprises mannose; and H of the peptide antigen conjugate comprises a poly(amino acid) containing a hydrophobic amino acid (M).

[0454] In some embodiments of the vaccine, S of the amphiphile comprises a second or third generation dendrimer; H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M); SG comprises mannose; and H of the peptide antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0455] In some embodiments of the vaccine, B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M); SG comprises mannose; and H of the peptide antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0456] In some embodiments of the vaccine, S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M); SG comprises mannose; and H of the peptide antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0457] In some embodiments of the vaccine, S of the amphiphile comprises a second or third generation dendrimer; H of the amphiphile comprises a polymer of p-aminophenylalanine; SG comprises mannose; and H of the peptide antigen conjugate comprises a poly(amino acid) containing a hydrophobic amino acid (M).

[0458] In some embodiments of the vaccine, B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a polymer of p-aminophenylalanine; SG comprises mannose; and H of the peptide antigen conjugate comprises a poly(amino acid) containing a hydrophobic amino acid (M).

[0459] In some embodiments of the vaccine, S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a polymer of p-aminophenylalanine; SG comprises mannose; and H of the peptide antigen conjugate comprises a poly(amino acid) containing a hydrophobic amino acid (M).

[0460] In some embodiments of the vaccine, S of the amphiphile comprises a second or third generation dendrimer; and H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and a reactive amino acid (N) containing an imidazoquinoline; SG comprises mannose; and H of the peptide antigen conjugate comprises a poly(amino acid) containing a hydrophobic amino acid (M).

[0461] In some embodiments of the vaccine, B comprises 4 to 36 PEG monomer units; and H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and a reactive amino acid (N) containing an imidazoquinoline; SG comprises mannose; and H of the peptide antigen conjugate comprises a poly(amino acid) containing a hydrophobic amino acid (M).

[0462] In some embodiments of the vaccine, S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and a reactive amino acid (N) containing an imidazoquinoline; SG comprises mannose; and H of the peptide antigen conjugate comprises a poly(amino acid) containing a hydrophobic amino acid (M).

[0463] In some embodiments of the vaccine, S of the amphiphile comprises a second or third generation dendrimer; and H of the amphiphile comprises a poly(amino acid) comprising tryptophan and an imidazoquinoline-reactive amino acid (N); SG comprises mannose; and H of the peptide antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0464] In some embodiments of the vaccine, B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a poly(amino acid) comprising tryptophan and an imidazoquinoline-reactive amino acid (N); SG comprises mannose; and H of the peptide antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0465] In some embodiments of the vaccine, S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a poly(amino acid) comprising tryptophan and an imidazoquinoline-reactive amino acid (N); SG comprises mannose; and H of the peptide antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0466] In some embodiments of the vaccine, S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; the amphiphile comprises amino-hexanoic acid; and H of the peptide antigen conjugate comprises a poly(amino acid) containing a hydrophobic amino acid (M).

[0467] In some embodiments of the vaccine, the S of the amphiphile comprises a second or third generation dendrimer; the H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M); the amphiphile comprises amino-hexanoic acid; and the H of the peptide antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0468] In some embodiments of the vaccine, B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M); the amphiphile comprises amino-hexanoic acid; and H of the peptide antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0469] In some embodiments of the vaccine, S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M); the amphiphile comprises amino-hexanoic acid; and H of the peptide antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0470] In some embodiments of the vaccine, S of the amphiphile comprises a second or third generation dendrimer; H of the amphiphile comprises a polymer of p-aminophenylalanine; the amphiphile comprises amino-hexanoic acid; and H of the peptide antigen conjugate comprises a poly(amino acid) containing a hydrophobic amino acid (M).

[0471] In some embodiments of the vaccine, B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a polymer of p-aminophenylalanine; the amphiphile comprises amino-hexanoic acid; and H of the peptide antigen conjugate comprises a poly(amino acid) containing a hydrophobic amino acid (M).

[0472] In some embodiments of the vaccine, S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a polymer of p-aminophenylalanine; the amphiphile comprises amino-hexanoic acid; and H of the peptide antigen conjugate comprises a poly(amino acid) containing a hydrophobic amino acid (M).

[0473] In some embodiments of the vaccine, S of the amphiphile comprises a second or third generation dendrimer; and H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and an imidazoquinoline-containing reactive amino acid (N); the amphiphile comprises amino-hexanoic acid; and H of the peptide antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0474] In some embodiments of the vaccine, B comprises 4 to 36 PEG monomer units; and H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and a reactive amino acid (N) containing an imidazoquinoline; the amphiphile comprises amino-hexanoic acid; and H of the peptide antigen conjugate comprises a poly(amino acid) containing a hydrophobic amino acid (M).

[0475] In some embodiments of the vaccine, S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and a reactive amino acid (N) containing an imidazoquinoline; the amphiphile comprises amino-hexanoic acid; and H of the peptide antigen conjugate comprises a poly(amino acid) containing a hydrophobic amino acid (M).

[0476] In some embodiments of the vaccine, S of the amphiphile comprises a second or third generation dendrimer; and H of the amphiphile comprises a poly(amino acid) comprising tryptophan and an imidazoquinoline-reactive amino acid (N); the amphiphile comprises amino-hexanoic acid; and H of the peptide antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0477] In some embodiments of the vaccine, B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises tryptophan and a poly(amino acid) containing an imidazoquinoline-reactive amino acid (N); the amphiphile comprises amino-hexanoic acid; and H of the peptide antigen conjugate comprises a poly(amino acid) containing a hydrophobic amino acid (M).

[0478] In some embodiments of the vaccine, S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a poly(amino acid) comprising tryptophan and an imidazoquinoline-reactive amino acid (N); the amphiphile comprises amino-hexanoic acid; and H of the peptide antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0479] In some embodiments of the vaccine, S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; the dendrimer monomers comprise hydroxy acids and amino alcohols; and H of the peptide antigen conjugate comprises a poly(amino acid) containing a hydrophobic amino acid (M).

[0480] In some embodiments of the vaccine, S of the amphiphile comprises a second or third generation dendrimer; H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M); the dendrimer monomers comprise a hydroxy acid and an amino alcohol; and H of the peptide antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0481] In some embodiments of the vaccine, B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M); the dendrimer monomer comprises a hydroxy acid and an amino alcohol; and H of the peptide antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0482] In some embodiments of the vaccine, S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M); the dendrimer monomers comprise hydroxy acids and amino alcohols; and H of the peptide antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0483] In some embodiments of the vaccine, S of the amphiphile comprises a second or third generation dendrimer; H of the amphiphile comprises a polymer of p-aminophenylalanine; the dendrimer monomers comprise hydroxy acids and amino alcohols; and H of the peptide antigen conjugate comprises a poly(amino acid) containing a hydrophobic amino acid (M).

[0484] In some embodiments of the vaccine, B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a polymer of p-aminophenylalanine; the dendrimer monomers comprise hydroxy acids and amino alcohols; and H of the peptide antigen conjugate comprises a poly(amino acid) containing a hydrophobic amino acid (M).

[0485] In some embodiments of the vaccine, S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a polymer of p-aminophenylalanine; the dendrimer monomers comprise hydroxy acids and amino alcohols; and H of the peptide antigen conjugate comprises a poly(amino acid) containing a hydrophobic amino acid (M).

[0486] In some embodiments of the vaccine, S of the amphiphile comprises a second or third generation dendrimer; and H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and a reactive amino acid (N) containing an imidazoquinoline; the dendrimer monomers comprise a hydroxy acid and an amino alcohol; and H of the peptide antigen conjugate comprises a poly(amino acid) containing a hydrophobic amino acid (M).

[0487] In some embodiments of the vaccine, B comprises 4 to 36 PEG monomer units; and H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and a reactive amino acid (N) containing an imidazoquinoline; the dendrimer monomer comprises a hydroxy acid and an amino alcohol; and H of the peptide antigen conjugate comprises a poly(amino acid) containing a hydrophobic amino acid (M).

[0488] In some embodiments of the vaccine, S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and a reactive amino acid (N) containing an imidazoquinoline; the dendrimer monomers comprise a hydroxy acid and an amino alcohol; and H of the peptide antigen conjugate comprises a poly(amino acid) containing a hydrophobic amino acid (M).

[0489] In some embodiments of the vaccine, S of the amphiphile comprises a second or third generation dendrimer; and H of the amphiphile comprises a poly(amino acid) comprising tryptophan and an imidazoquinoline-reactive amino acid (N); the dendrimer monomers comprise a hydroxy acid and an amino alcohol; and H of the peptide antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0490] In some embodiments of the vaccine, B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a poly(amino acid) comprising tryptophan and an imidazoquinoline-reactive amino acid (N); the dendrimer monomer comprises a hydroxy acid and an amino alcohol; and H of the peptide antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0491] In some embodiments of the vaccine, S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a poly(amino acid) comprising tryptophan and an imidazoquinoline-reactive amino acid (N); the dendrimer monomers comprise a hydroxy acid and an amino alcohol; and H of the peptide antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0492] The present disclosure relates to a vaccine for inducing tolerance, comprising at least one peptide antigen conjugate having a formula selected from the group consisting of PEG-[E1]-A-[E2]-[U]-H and H-[U]-[E1]-A-[E2]-PEG

[0493] in

[0494] A is a peptide antigen;

[0495] E1 is the N-terminal extension;

[0496] E2 is the C-terminal extension;

[0497] H is independently a hydrophobic block at each occurrence, wherein one or more drug molecules (D) are optionally attached to each H directly or via a suitable linker X1;

[0498] U is independently a linker at each occurrence;

[0499] [ ] indicates that the group is optional, and

[0500] - denotes that two adjacent groups are directly attached to each other by a covalent bond or indirectly attached to each other via a suitable linker X; and wherein the at least one peptide antigen is selected from the group consisting of an autoantigen, an alloantigen and an allergen.

[0501] In some embodiments of the vaccine, wherein (i) the average aqueous solubility of the peptide antigen (A) of the one or more peptide antigen conjugates is less than 1 mg / mL, or (ii) the average GRAVY score of the peptide antigen (A) of the one or more peptide antigen conjugates is >0, the amphiphile is present.

[0502] The present disclosure also relates to a vaccine for inducing tolerance, comprising at least one peptide antigen conjugate having a formula selected from the group consisting of PEG-[E1]-A-[E2]-[U]-H and H-[U]-[E1]-A-[E2]-PEG, and an amphiphile having the formula S-[B]-[U]-H,

[0503] in

[0504] A is a peptide antigen;

[0505] E1 is the N-terminal extension;

[0506] E2 is the C-terminal extension;

[0507] H is independently a hydrophobic block at each occurrence, wherein one or more drug molecules (D) are optionally attached to each H directly or via a suitable linker X1;

[0508] S is the solubilization block;

[0509] B is a spacer;

[0510] U is independently a linker at each occurrence;

[0511] [ ] indicates that the group is optional,

[0512] - means that two adjacent groups are directly attached to each other by a covalent bond or indirectly attached to each other via a suitable linker X; and

[0513] wherein the amphiphile comprises a dendrimer amplifier and the at least one peptide antigen is selected from the group consisting of an autoantigen, an alloantigen, and an allergen.

[0514] In one embodiment of the vaccine, the PEG group of the peptide antigen conjugate includes a terminal functional group selected from OH, MeO- and NH2.

[0515] In a specific embodiment of a vaccine for inducing tolerance, the PEG group of the peptide antigen conjugate is polyethylene glycol.

[0516] In some embodiments of the vaccine for inducing tolerance, the PEG group of the peptide antigen conjugate comprises between 4 and 36 monomer units.

[0517] In specific embodiments of the vaccine for inducing tolerance, the PEG group of the peptide antigen conjugate comprises between 4 and 12 monomeric units, or between 12 and 36 monomeric units, preferably 24 monomeric units.

[0518] In some embodiments of the vaccine for inducing tolerance, A is a self-antigen. In other embodiments, A is an allergen. In other embodiments, A is an allergen.

[0519] In some embodiments of the vaccine for inducing tolerance, wherein the amphiphile is present, the amphiphile comprises a dendrimer amplifier.

[0520] In some embodiments of the vaccine for inducing tolerance, the amphiphile S comprises a dendrimer amplifier. In other embodiments, the amphiphile S has a dendritic architecture.

[0521] In some embodiments of vaccines for inducing tolerance, the S of the amphiphile comprises two or more solubilizing groups (SG). In other embodiments, two or more SGs are linked to the rest of S via a dendrimer amplifier, for example, 4 to 8 SGs are linked to S.

[0522] In some embodiments of the vaccine for inducing tolerance, SG is independently selected from amines, hydroxyls, carboxylic acids and / or sugar molecules, wherein the sugar molecules are independently selected from mannose, glucose, glucosamine, N-acetylglucose, galactose, galactosamine, N-acetylgalactosamine, N-acetylglucosamine, phosphoserine and any derivatives thereof, agonists of CD22a, sialyl Lewis x, and combinations thereof.

[0523] In some embodiments of the vaccine for inducing tolerance, at least one SG is galactose. In other embodiments, at least one SG is phosphoserine. In other embodiments, at least one SG is an agonist of CD22a.

[0524] In some embodiments of vaccines for inducing tolerance, the dendrimer amplifier comprises 1 to 10 generations of repeating monomer units, each generation having between 2 and 6 branches. In other embodiments, the dendrimer amplifier comprises 2 to 3 generations of repeating monomer units, each generation having between 2 and 3 branches.

[0525] In some embodiments of the vaccine for inducing tolerance, the repeating monomer unit is selected from FG1-(CH2) y2 CH(R1 )2、FG1-(CH2) y2 C(R 1 )3、FG1-(CH2CH2O) y2 CH(R 1 )2、FG1-(CH2CH2O) y2 C(R 1 )3、and FG1-CH(R 1 )2、FG1-C(R 1 )3, where R 1 is independently selected at each occurrence from (CH2) y3 -FG2, (OCH2CH2) y3 -FG2 and CH2(OCH2CH2) y3 -FG2); y2 and y3 are independently an integer of repeating units from 1 to 6 at each occurrence; FG1 is a first functional group; and FG2 is a second functional group. In some embodiments, FG1 is -NH2; and FG2 is independently -CO2- or -CO2H at each occurrence. In some embodiments, FG1 is -CO2- or -CO2H; and FG2 is independently -NH2 at each occurrence.

[0526] In some embodiments of the vaccine for inducing tolerance, SG is linked to S via a suitable linker X5. In some embodiments of the vaccine for inducing tolerance, the suitable linker X5 linking SG to S is selected from a lower alkyl group and a PEG group. In some embodiments of the vaccine for inducing tolerance, two or more SG are linked to the rest of S via a dendrimer amplifier via a suitable linker X5, which links the two or more SG to the terminal functional group (FGt) of the dendrimer amplifier via an amide bond. In some embodiments of the vaccine for inducing tolerance, the linker X5 linking SG to the dendrimer amplifier is selected from a -NH-R 19 、-NH-C(O)-R 19 、-C(O)-NH-R 19 -or-C(O)-R 19 , where R 19 Can be selected from but not limited to -(CH2) t -、-(CH2CH2O) t -CH2CH2-, -(CH2)tC(O)-NH-(CH2) u -、-(CH2CH2O) t CH2CH2C(O)-NH-(CH2) u -、-(CH2) t -NH-C(O)-NH-(CH2) u-or-(CH2CH2O) t CH2CH2NH-C(O)-(CH2) u -, wherein t and u are each independently an integer generally selected from between 1 and 6, such as 1, 2, 3, 4, 5 or 6.

[0527] In some embodiments of vaccines for inducing tolerance, the dendrimer amplifier comprises a polyethylene oxide (PEG) group.

[0528] In some embodiments of the vaccine for inducing tolerance, H of the amphiphile comprises a higher alkane, an aromatic group, a fatty acid, a sterol, a polyunsaturated hydrocarbon, squalene, a saponin, and / or a polymer.

[0529] In some embodiments of the vaccine for inducing tolerance, H of the peptide antigen conjugate comprises a higher alkane, an aromatic group, a fatty acid, a sterol, a polyunsaturated hydrocarbon, and / or a polymer.

[0530] In some embodiments of the vaccine for inducing tolerance, each H independently comprises a poly(amino acid) comprising a monomer selected from the group consisting of a hydrophobic amino acid (M), a reactive amino acid (N), a spacer amino acid (O), a charged amino acid (P), and combinations thereof, with the proviso that at least one of M or N is present.

[0531] In some embodiments of the vaccine for inducing tolerance, each H independently comprises a poly(amino acid) having the formula:

[0532] ,

[0533] wherein M, N, O and P are each independently present or absent, provided that at least one of M or N is present;

[0534] m, n, o and p each independently represent an integer from 1 to 100, and the sum of m, n, o and p is less than or equal to 100;

[0535] R 3 Selected from hydrogen, NH2, NH-CH3, NH-(CH2) y5 CH3, OH or a drug molecule (D) directly or via a suitable linker X1; and

[0536] y5 is an integer selected from 1 to 6.

[0537] In some embodiments of vaccines for inducing tolerance, P is absent. In other embodiments, N, O, and P are each absent.

[0538] In some embodiments of vaccines for inducing tolerance, P is , where each R5 Independently, a group comprising 1 to 2 charged functional groups.

[0539] In some embodiments of the vaccine for inducing tolerance, O is , wherein each Q is independently selected from (CH2) y and (CH2CH2O) i CH2CH2; each y is independently selected from an integer from 1 to 6; and each i is independently selected from an integer from 1 to 4.

[0540] In some embodiments of vaccines for inducing tolerance, N is , wherein each X1 is independently a suitable linker; and each D is independently a drug molecule.

[0541] In some embodiments of the vaccine for inducing tolerance, M is , where each R 4 are independently hydrophobic groups.

[0542] In some embodiments of vaccines for inducing tolerance, R 4 yes

[0543]

[0544] in

[0545] α is aryl or heteroaryl;

[0546] X2 is present or absent and, when present, is a suitable linker;

[0547] Y8 is an integer selected from 0 and 6; and

[0548] Z 1 、Z 2 and Z 3 Each is independently selected from H, F, hydroxy, amino, alkyl and fluoroalkyl.

[0549] In some embodiments of vaccines for inducing tolerance, α is an aryl group, e.g., phenyl or naphthyl. In other embodiments, α is a heteroaryl group, e.g., pyridyl, quinolyl, isoquinolyl, indolyl, or benzimidazolyl.

[0550] In some embodiments of vaccines for inducing tolerance, X is absent. In other embodiments, X2 is present and is selected from C(O), CO2(CH2) y9 、CO2、C(O)NH(CH2) y9 , NHC(O) and NHC(O)(CH2) y9，wherein y9 is an integer generally selected from 1 to 6. In other embodiments, X2 is present and is selected from lower alkyl and PEG groups.

[0551] In some embodiments of vaccines for inducing tolerance, each R 4 Independently selected from:

[0552] 、 、 、

[0553] 、 、 、

[0554] 、 、 、

[0555] 、 、 、

[0556] and ,

[0557] wherein each X2 is independently selected from a suitable linker, and each y8 is independently selected from an integer between 0 and 6. In other embodiments, each R 4 Independently selected from:

[0558] 、 、 、

[0559] 、 、 、

[0560] 、 、 、

[0561] 、 、 、

[0562] and ,

[0563] wherein each y8 is independently selected from an integer between 0 and 6. In other embodiments, each R 4 Independently selected from:

[0564] 、 、 、 、 、 、 、 、 、 、 、 and In other embodiments, each R 4 Independently selected from:

[0565] 、 、 、

[0566] 、 、

[0567] 、 、 、

[0568] 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 and In a preferred embodiment, each R 4 Independently selected from:

[0569] 、 、 and , wherein y is selected from an integer between 1 and 6.

[0570] In some embodiments of the vaccine for inducing tolerance, wherein at least one D is:

[0571]

[0572] in,

[0573] R 20is selected from the group consisting of H, alkyl, alkoxyalkyl, aryl, heteroaryl, aminoalkyl, amide, and ester; and X3 is selected from the group consisting of alkyl, alkoxyalkyl, aralkyl, heteroaralkyl, aryl, heteroaryl, and carboxyl.

[0574] In some embodiments of vaccines for inducing tolerance, R 20 is selected from H, alkyl and alkoxyalkyl; and X3 is selected from alkyl and aralkyl. In other embodiments, R 20 It's butyl.

[0575] In some embodiments of the vaccine for inducing tolerance, X3 is alkyl.

[0576] In some embodiments of the vaccine for inducing tolerance, m, n, o, and p each independently represent an integer from 1 to 30, and the sum of m, n, o, and p is less than or equal to 30.

[0577] In some embodiments of the vaccine for inducing tolerance, m, n, o, and p each independently represent an integer from 1 to 10, and the sum of m, n, o, and p is less than or equal to 10.

[0578] In some embodiments of vaccines for inducing tolerance, B is present and is a hydrophilic polymer, e.g., a PEG group. In other embodiments, B is present and is a hydrophilic peptide.

[0579] In some embodiments of vaccines for inducing tolerance, the PEG group comprises between 4 and 36 monomeric units. In other embodiments, the PEG group comprises between 4 and 12 monomeric units.

[0580] In some embodiments of the vaccine for inducing tolerance, the hydrophilic peptide comprises between 4 and 36 amino acids. In other embodiments, the hydrophilic peptide comprises between 4 and 12 amino acids.

[0581] In some embodiments of vaccines for inducing tolerance, the amphiphile has the formula SH. In other embodiments, the amphiphile has the formula SBUH. In other embodiments, the amphiphile has the formula SBUHD.

[0582] In some embodiments of the vaccine for inducing tolerance, the vaccine comprises a molar ratio of peptide antigen conjugate to amphiphile of between about 4:1 to about 1:20, preferably about 1:1.

[0583] In some embodiments of the vaccine for inducing tolerance, the peptide antigen (A) comprises a sequence in which one or more cysteine ​​residues have been replaced by α-aminobutyric acid and / or one or more methionine residues have been replaced by norleucine.

[0584] In some embodiments of the vaccine for inducing tolerance, the peptide antigen (A) comprises alpha-aminobutyric acid and / or norleucine.

[0585] In some embodiments of the vaccine for inducing tolerance, the vaccine comprises at least one D selected from the group consisting of an inhibitor of mTOR, RORγt, CDK8 / 19, and HDAC, and an inhibitor of AHR, RAR, and A 2a In other embodiments, the at least one D is selected from ATP-competitive mTOR inhibitors.

[0586] In some embodiments of the vaccine for inducing tolerance, the vaccine further comprises a second drug molecule (D2) independently selected from the group consisting of inhibitors of mTOR, RORγt, CDK8 / 19 and HDAC; inhibitors of AHR, RAR and A 2a and an immunostimulatory agent selected from the group consisting of an agonist of NLR, CLR, TLR, and STING, with the proviso that D and D2 bind to different receptors.

[0587] In some embodiments of the vaccine for inducing tolerance, the at least one D is selected from an inhibitor of mTOR and an agonist of AHR, and the D2 is selected from an agonist of NLR, CLR, TLR, and STING.

[0588] In some embodiments of the vaccine for inducing tolerance, wherein the at least one D is selected from ATP-competitive mTOR inhibitors, and the D2 is selected from agonists of NLR, CLR, TLR, and STING.

[0589] In other embodiments of the vaccine for inducing tolerance, the D2 is selected from agonists of TLR-3, TLR-7, TLR-8, TLR-7 / 8, TLR-9 and STING. In other embodiments of the vaccine for inducing tolerance, the D2 is selected from RNA and imidazoquinoline agonists of TLR-7, TLR-8 and TLR-7 / 8.

[0590] In some embodiments of the vaccine for inducing tolerance, the vaccine further comprises a third drug molecule (D3) independently selected from the group consisting of inhibitors of mTOR, RORγt, CDK8 / 19 and HDAC; inhibitors of AHR, RAR and A 2a and an immunostimulatory agent selected from the group consisting of an agonist of NLR, CLR, TLR and STING, with the proviso that D, D2 and D3 bind to different receptors.

[0591] In some embodiments of the vaccine for inducing tolerance, the at least one D is selected from AZD-8055, AZD2016, KU-0063794, CC223, Torin-1, Torin-2, INK-128, WYE354, WYE132, OSI-027, OXA-01, PI-103, NVP-BEZ235, GNE-493, GSK2126458, rapamycin, tacrolimus, everolimus, RAD001, CCI-779 and AP23573.

[0592] In some embodiments of the vaccine for inducing tolerance, the molar ratio of total peptide antigen conjugate to at least one D is between about 20:1 and 1:2, or between about 10:1 and about 1:1, or between about 4:1 and about 2:1, or preferably about 1:1.

[0593] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; and B comprises 4 to 36 PEG monomer units.

[0594] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; and H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0595] In some embodiments of the vaccine for inducing tolerance, B comprises 4 to 36 PEG monomer units; and H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0596] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; and H of the amphiphile comprises a poly(amino acid) containing a hydrophobic amino acid (M).

[0597] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; and H of the amphiphile comprises a polymer of p-aminophenylalanine.

[0598] In some embodiments of the vaccine for inducing tolerance, B comprises 4 to 36 PEG monomer units; and H of the amphiphile comprises a polymer of p-aminophenylalanine.

[0599] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; and H of the amphiphile comprises a polymer of p-aminophenylalanine.

[0600] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; and H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and an imidazoquinoline-containing reactive amino acid (N).

[0601] In some embodiments of the vaccine for inducing tolerance, B comprises 4 to 36 PEG monomer units; and H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and an imidazoquinoline-containing reactive amino acid (N).

[0602] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; and H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and an imidazoquinoline-containing reactive amino acid (N).

[0603] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; and H of the amphiphile comprises a poly(amino acid) of tryptophan and an imidazoquinoline-reactive amino acid (N).

[0604] In some embodiments of the vaccine for inducing tolerance, B comprises 4 to 36 PEG monomer units; and H of the amphiphile comprises a poly(amino acid) of tryptophan and an imidazoquinoline-containing reactive amino acid (N).

[0605] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; and H of the amphiphile comprises a poly(amino acid) of tryptophan and an imidazoquinoline-reactive amino acid (N).

[0606] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; and SG comprises N-acetylgalactosamine.

[0607] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M); and SG comprises N-acetylgalactosamine.

[0608] In some embodiments of the vaccine for inducing tolerance, B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M); and SG comprises N-acetylgalactosamine.

[0609] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a poly(amino acid) containing a hydrophobic amino acid (M); and SG comprises N-acetylgalactosamine.

[0610] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; H of the amphiphile comprises a polymer of p-aminophenylalanine; and SG comprises N-acetylgalactosamine.

[0611] In some embodiments of the vaccine for inducing tolerance, B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a polymer of p-aminophenylalanine; and SG comprises N-acetylgalactosamine.

[0612] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a polymer of p-aminophenylalanine; and SG comprises N-acetylgalactosamine.

[0613] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; and H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and an imidazoquinoline-containing reactive amino acid (N); and SG comprises N-acetylgalactosamine.

[0614] In some embodiments of the vaccine for inducing tolerance, B comprises 4 to 36 PEG monomer units; and H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and an imidazoquinoline-containing reactive amino acid (N); and SG comprises N-acetylgalactosamine.

[0615] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and an imidazoquinoline-containing reactive amino acid (N); and SG comprises N-acetylgalactosamine.

[0616] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; and H of the amphiphile comprises a poly(amino acid) of tryptophan and an imidazoquinoline-reactive amino acid (N); and SG comprises N-acetylgalactosamine.

[0617] In some embodiments of the vaccine for inducing tolerance, B comprises 4 to 36 PEG monomer units; and H of the amphiphile comprises a poly(amino acid) of tryptophan and an imidazoquinoline-containing reactive amino acid (N); and SG comprises N-acetylgalactosamine.

[0618] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; and H of the amphiphile comprises a poly(amino acid) of tryptophan and an imidazoquinoline-reactive amino acid (N); and SG comprises N-acetylgalactosamine.

[0619] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; and B comprises 4 to 36 PEG monomer units; and the peptide antigen conjugate comprises an enzymatically degradable linker.

[0620] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; and H of the amphiphile comprises a poly(amino acid) containing a hydrophobic amino acid (M); and the peptide antigen conjugate comprises an enzymatically degradable linker.

[0621] In some embodiments of the vaccine for inducing tolerance, B comprises 4 to 36 PEG monomer units; and H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M); and the peptide antigen conjugate comprises an enzymatically degradable linker.

[0622] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a poly(amino acid) containing a hydrophobic amino acid (M); and the peptide antigen conjugate comprises an enzymatically degradable linker.

[0623] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; and H of the amphiphile comprises a polymer of p-aminophenylalanine; and the peptide antigen conjugate comprises an enzymatically degradable linker.

[0624] In some embodiments of the vaccine for inducing tolerance, B comprises 4 to 36 PEG monomer units; and H of the amphiphile comprises a polymer of p-aminophenylalanine; and the peptide antigen conjugate comprises an enzymatically degradable linker.

[0625] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; and H of the amphiphile comprises a polymer of p-aminophenylalanine; and the peptide antigen conjugate comprises an enzymatically degradable linker.

[0626] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; and H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and an imidazoquinoline-containing reactive amino acid (N); and the peptide antigen conjugate comprises an enzymatically degradable linker.

[0627] In some embodiments of the vaccine for inducing tolerance, B comprises 4 to 36 PEG monomer units; and H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and an imidazoquinoline-containing reactive amino acid (N); and the peptide antigen conjugate comprises an enzyme-degradable linker.

[0628] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; and H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and an imidazoquinoline-containing reactive amino acid (N); and the peptide antigen conjugate comprises an enzymatically degradable linker.

[0629] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; and H of the amphiphile comprises a poly(amino acid) of tryptophan and an imidazoquinoline-reactive amino acid (N); and the peptide antigen conjugate comprises an enzymatically degradable linker.

[0630] In some embodiments of the vaccine for inducing tolerance, B comprises 4 to 36 PEG monomer units; and H of the amphiphile comprises a poly(amino acid) of tryptophan and an imidazoquinoline-containing reactive amino acid (N); and the peptide antigen conjugate comprises an enzymatically degradable linker.

[0631] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; and H of the amphiphile comprises a poly(amino acid) of tryptophan and an imidazoquinoline-reactive amino acid (N); and the peptide antigen conjugate comprises an enzymatically degradable linker.

[0632] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; and H of the peptide antigen conjugate comprises a poly(amino acid) containing a hydrophobic amino acid (M).

[0633] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M); and H of the peptide antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0634] In some embodiments of the vaccine for inducing tolerance, B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M); and H of the peptide antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0635] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M); and H of the peptide antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0636] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; H of the amphiphile comprises a polymer of p-aminophenylalanine; and H of the peptide antigen conjugate comprises a poly(amino acid) containing a hydrophobic amino acid (M).

[0637] In some embodiments of the vaccine for inducing tolerance, B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a polymer of p-aminophenylalanine; and H of the peptide antigen conjugate comprises a poly(amino acid) containing a hydrophobic amino acid (M).

[0638] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a polymer of p-aminophenylalanine; and H of the peptide antigen conjugate comprises a poly(amino acid) containing a hydrophobic amino acid (M).

[0639] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and an imidazoquinoline-containing reactive amino acid (N); and H of the peptide antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0640] In some embodiments of the vaccine for inducing tolerance, B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and a reactive amino acid (N) containing an imidazoquinoline; and H of the peptide antigen conjugate comprises a poly(amino acid) containing a hydrophobic amino acid (M).

[0641] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and an imidazoquinoline-containing reactive amino acid (N); and H of the peptide antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0642] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; H of the amphiphile comprises a poly(amino acid) comprising tryptophan and an imidazoquinoline-reactive amino acid (N); and H of the peptide antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0643] In some embodiments of the vaccine for inducing tolerance, B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a poly(amino acid) comprising tryptophan and an imidazoquinoline-reactive amino acid (N); and H of the peptide antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0644] In some embodiments of the vaccine for inducing tolerance, wherein S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a poly(amino acid) comprising tryptophan and an imidazoquinoline-reactive amino acid (N); and H of the peptide antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0645] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; SG comprises N-acetylgalactosamine; and H of the peptide antigen conjugate comprises a poly(amino acid) containing a hydrophobic amino acid (M).

[0646] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M); SG comprises N-acetylgalactosamine; and H of the peptide antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0647] In some embodiments of the vaccine for inducing tolerance, B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M); SG comprises N-acetylgalactosamine; and H of the peptide antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0648] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M); SG comprises N-acetylgalactosamine; and H of the peptide antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0649] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; H of the amphiphile comprises a polymer of p-aminophenylalanine; SG comprises N-acetylgalactosamine; and H of the peptide antigen conjugate comprises a poly(amino acid) containing a hydrophobic amino acid (M).

[0650] In some embodiments of the vaccine for inducing tolerance, B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a polymer of p-aminophenylalanine; SG comprises N-acetylgalactosamine; and H of the peptide antigen conjugate comprises a poly(amino acid) containing a hydrophobic amino acid (M).

[0651] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a polymer of p-aminophenylalanine; SG comprises N-acetylgalactosamine; and H of the peptide antigen conjugate comprises a poly(amino acid) containing a hydrophobic amino acid (M).

[0652] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and a reactive amino acid (N) containing an imidazoquinoline; and SG comprises N-acetylgalactosamine; and H of the peptide antigen conjugate comprises a poly(amino acid) containing a hydrophobic amino acid (M).

[0653] In some embodiments of the vaccine for inducing tolerance, B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and a reactive amino acid (N) containing an imidazoquinoline; SG comprises N-acetylgalactosamine; and H of the peptide antigen conjugate comprises a poly(amino acid) containing a hydrophobic amino acid (M).

[0654] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and a reactive amino acid (N) containing an imidazoquinoline; SG comprises N-acetylgalactosamine; and H of the peptide antigen conjugate comprises a poly(amino acid) containing a hydrophobic amino acid (M).

[0655] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; H of the amphiphile comprises a poly(amino acid) comprising tryptophan and an imidazoquinoline-reactive amino acid (N); SG comprises N-acetylgalactosamine; and H of the peptide antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0656] In some embodiments of the vaccine for inducing tolerance, B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a poly(amino acid) comprising tryptophan and an imidazoquinoline-reactive amino acid (N); SG comprises N-acetylgalactosamine; and H of the peptide antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0657] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a poly(amino acid) comprising tryptophan and an imidazoquinoline-reactive amino acid (N); and SG comprises N-acetylgalactosamine; and H of the peptide antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0658] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; the peptide antigen conjugate comprises an enzymatically degradable linker; and H of the peptide antigen conjugate comprises a poly(amino acid) containing a hydrophobic amino acid (M).

[0659] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; and H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M); the peptide antigen conjugate comprises an enzymatically degradable linker; and H of the peptide antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0660] In some embodiments of the vaccine for inducing tolerance, B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M); the peptide antigen conjugate comprises an enzymatically degradable linker; and H of the peptide antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0661] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; the amphiphile comprises amino-hexanoic acid; and H of the peptide antigen conjugate comprises a poly(amino acid) containing a hydrophobic amino acid (M).

[0662] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M); the amphiphile comprises amino-hexanoic acid; and H of the peptide antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0663] In some embodiments of the vaccine for inducing tolerance, B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M); the amphiphile comprises amino-hexanoic acid; and H of the peptide antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0664] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M); the amphiphile comprises amino-hexanoic acid; and H of the peptide antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0665] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; H of the amphiphile comprises a polymer of p-aminophenylalanine; the amphiphile comprises amino-hexanoic acid; and H of the peptide antigen conjugate comprises a poly(amino acid) containing a hydrophobic amino acid (M).

[0666] In some embodiments of the vaccine for inducing tolerance, B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a polymer of p-aminophenylalanine; the amphiphile comprises amino-hexanoic acid; and H of the peptide antigen conjugate comprises a poly(amino acid) containing a hydrophobic amino acid (M).

[0667] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a polymer of p-aminophenylalanine; the amphiphile comprises amino-hexanoic acid; and H of the peptide antigen conjugate comprises a poly(amino acid) containing a hydrophobic amino acid (M).

[0668] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and a reactive amino acid (N) containing an imidazoquinoline; the amphiphile comprises amino-hexanoic acid; and H of the peptide antigen conjugate comprises a poly(amino acid) containing a hydrophobic amino acid (M).

[0669] In some embodiments of the vaccine for inducing tolerance, B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and a reactive amino acid (N) containing an imidazoquinoline; the amphiphile comprises amino-hexanoic acid; and H of the peptide antigen conjugate comprises a poly(amino acid) containing a hydrophobic amino acid (M).

[0670] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and a reactive amino acid (N) containing an imidazoquinoline; the amphiphile comprises amino-hexanoic acid; and H of the peptide antigen conjugate comprises a poly(amino acid) containing a hydrophobic amino acid (M).

[0671] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; H of the amphiphile comprises a poly(amino acid) comprising tryptophan and an imidazoquinoline-reactive amino acid (N); the amphiphile comprises amino-hexanoic acid; and H of the peptide antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0672] In some embodiments of the vaccine for inducing tolerance, B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises tryptophan and a poly(amino acid) containing an imidazoquinoline-reactive amino acid (N); the amphiphile comprises amino-hexanoic acid; and H of the peptide antigen conjugate comprises a poly(amino acid) containing a hydrophobic amino acid (M).

[0673] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a poly(amino acid) comprising tryptophan and an imidazoquinoline-reactive amino acid (N); the amphiphile comprises amino-hexanoic acid; and H of the peptide antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0674] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; the dendrimer monomers comprise hydroxy acids and amino alcohols; and H of the peptide antigen conjugate comprises a poly(amino acid) containing a hydrophobic amino acid (M).

[0675] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M); the dendrimer monomers comprise a hydroxy acid and an amino alcohol; and H of the peptide antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0676] In some embodiments of the vaccine for inducing tolerance, B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M); the dendrimer monomer comprises a hydroxy acid and an amino alcohol; and H of the peptide antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0677] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M); the dendrimer monomers comprise a hydroxy acid and an amino alcohol; and H of the peptide antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0678] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; H of the amphiphile comprises a polymer of p-aminophenylalanine; the dendrimer monomers comprise hydroxy acids and amino alcohols; and H of the peptide antigen conjugate comprises a poly(amino acid) containing a hydrophobic amino acid (M).

[0679] In some embodiments of the vaccine for inducing tolerance, B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a polymer of p-aminophenylalanine; the dendrimer monomers comprise a hydroxy acid and an amino alcohol; and H of the peptide antigen conjugate comprises a poly(amino acid) containing a hydrophobic amino acid (M).

[0680] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a polymer of p-aminophenylalanine; the dendrimer monomers comprise hydroxy acids and amino alcohols; and H of the peptide antigen conjugate comprises a poly(amino acid) containing a hydrophobic amino acid (M).

[0681] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and a reactive amino acid (N) containing an imidazoquinoline; the dendrimer monomer comprises a hydroxy acid and an amino alcohol; and H of the peptide antigen conjugate comprises a poly(amino acid) containing a hydrophobic amino acid (M).

[0682] In some embodiments of the vaccine for inducing tolerance, B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and a reactive amino acid (N) containing an imidazoquinoline; the dendrimer monomer comprises a hydroxy acid and an amino alcohol; and H of the peptide antigen conjugate comprises a poly(amino acid) containing a hydrophobic amino acid (M).

[0683] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and a reactive amino acid (N) containing an imidazoquinoline; the dendrimer monomers comprise a hydroxy acid and an amino alcohol; and H of the peptide antigen conjugate comprises a poly(amino acid) containing a hydrophobic amino acid (M).

[0684] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; H of the amphiphile comprises a poly(amino acid) comprising tryptophan and an imidazoquinoline-reactive amino acid (N); the dendrimer monomer comprises a hydroxy acid and an amino alcohol; and H of the peptide antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0685] In some embodiments of the vaccine for inducing tolerance, B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a poly(amino acid) comprising tryptophan and an imidazoquinoline-reactive amino acid (N); the dendrimer monomer comprises a hydroxy acid and an amino alcohol; and H of the peptide antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0686] In some embodiments of the vaccine for inducing tolerance, S of the amphiphile comprises a second or third generation dendrimer; B comprises 4 to 36 PEG monomer units; H of the amphiphile comprises a poly(amino acid) comprising tryptophan and an imidazoquinoline-containing reactive amino acid (N); the dendrimer monomers comprise a hydroxy acid and an amino alcohol; and H of the peptide antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0687] The present disclosure also relates to a vaccine comprising at least one peptide antigen conjugate having a formula selected from the group consisting of PEG-[E1]-A-[E2]-[U]-H and H-[U]-[E1]-A-[E2]-PEG, and an amphiphile having the formula S-[B]-[U]-H,

[0688] in

[0689] A is a peptide antigen;

[0690] E1 is the N-terminal extension;

[0691] E2 is the C-terminal extension;

[0692] H is independently a hydrophobic block at each occurrence, wherein one or more drug molecules (D) are optionally attached to each H directly or via a suitable linker X1;

[0693] S is the solubilization block;

[0694] B is a spacer;

[0695] U is independently a linker at each occurrence;

[0696] [ ] indicates that the group is optional,

[0697] - means that two adjacent groups are directly attached to each other by a covalent bond or indirectly attached to each other via a suitable linker X;

[0698] wherein the amphiphile comprises a dendrimer amplifier; and at least one A comprises a sequence in which one or more cysteine ​​residues have been replaced by α-aminobutyric acid and / or one or more methionine residues have been replaced by norleucine.

[0699] In some embodiments of the vaccine, S of the amphiphile comprises a carboxylic acid. In other embodiments of the vaccine, S of the amphiphile comprises succinic acid or beta-alanine.

[0700] In some embodiments of the vaccine, the molar ratio of peptide antigen conjugate to amphiphile is between about 4:1 and 1:20, preferably about 1:1.

[0701] In some embodiments of the vaccine, the average net charge of the at least one peptide antigen conjugate is positive at physiological pH, and the molar ratio of the peptide antigen conjugate to the amphiphile is between about 4:1 and about 2:1, or between about 1:2 and about 1:16, or between about 1:2 and about 1:4, preferably about 1:1.

[0702] The present disclosure also relates to a vaccine comprising at least one peptide antigen (A), wherein at least one peptide antigen (A) comprises a sequence in which one or more cysteine ​​residues have been replaced by α-aminobutyric acid and / or one or more methionine residues have been replaced by norleucine.

[0703] In some embodiments of the vaccine, the vaccine further comprises a particle delivery system selected from the group consisting of lipid emulsions, liposomes, PLGA particles, inorganic salt particles, and metal nanoparticles. In other embodiments of the vaccine, the vaccine further comprises at least one drug molecule (D) selected from the group consisting of immunostimulants and Treg-promoting immunomodulators.

[0704] The present disclosure also relates to a vaccine comprising at least one peptide antigen conjugate having a formula selected from the group consisting of PEG-[E1]-A-[E2]-[U]-H and H-[U]-[E1]-A-[E2]-PEG,

[0705] in

[0706] H is independently a hydrophobic block at each occurrence,

[0707] wherein one or more drug molecules (D) are optionally attached to each H directly or via a suitable linker X1;

[0708] A is independently a peptide antigen at each occurrence;

[0709] E1 is independently an N-terminal extension at each occurrence;

[0710] E2 is independently a C-terminal extension at each occurrence;

[0711] U is independently a linker at each occurrence;

[0712] in:

[0713] (i) at least one A comprises α-aminobutyric acid and / or norleucine;

[0714] (ii) at least one A is selected from a tumor antigen, at least one D is present and selected from an agonist of TLR-7 / 8, and the vaccine further comprises a second drug molecule (D2) selected from an inhibitor of mTOR;

[0715] (iii) at least one A is a glycopeptide; or

[0716] (iv) at least one A is selected from the group consisting of an autoantigen, an allergen, and an alloantigen, and at least one D is present and selected from the group consisting of an ATP-competitive mTOR inhibitor; [ ] indicates that the group is optional; and

[0717] – denotes that two adjacent groups are directly attached to each other by a covalent bond or indirectly attached to each other via a suitable linker X.

[0718] In some embodiments of the vaccine, at least one peptide antigen conjugate comprises at least one A selected from a tumor antigen.

[0719] In some embodiments of the vaccine, at least one D is selected from agonists of TLR-3, TLR-7, TLR-8, TLR-9, and STING.

[0720] In some embodiments of the vaccine, each H of the amphiphile and / or peptide antigen conjugate comprises a poly(amino acid) of a monomer comprising a reactive amino acid (N), wherein the monomer comprises a D selected from an agonist of TLR-7 / 8.

[0721] In some embodiments of the vaccine, D2 is present and is selected from rapamycin, tacrolimus, everolimus, RAD001, CCI-779, and AP23573.

[0722] In some embodiments of the vaccine, the molar ratio of the peptide antigen conjugate to D2 is between about 20:1 and 1:2, or between about 10:1 and about 1:1, or between about 4:1 and about 2:1, preferably about 1:1.

[0723] In some embodiments of the vaccine, at least one A is a glycopeptide, for example, A is a glycopeptide selected from the group consisting of HGVT*S*APDT*RPAPGS*T*APPA (SEQ ID NO: 534), DT*RPAPGS*T*APPAHGVT*S*AP (SEQ ID NO: 535), GS*T*APPAHGVT*S*APDT*RPAPGS*T*APPA (SEQ ID NO: 536), GVT*S*APDT*RPAP (SEQ ID NO: 537), APDT*RPAPGS*T*A (SEQ ID NO: 538), GS*T*APPAHGVT*S*AP (SEQ ID NO: 539), VT*S*AP (SEQ ID NO: 540), DT*RPAP (SEQ ID NO: 541), and GS*T*AP (SEQ ID NO: 542). 542), wherein * is an O-linked glycan and is independently selected at each occurrence from sialyl Lewis x, sialyl Lewis a, Lewis y, Lewis x, Tn, sTn, TF, sTF.

[0724] In some embodiments of the vaccine where A is a glycopeptide, S is absent. In other embodiments, S is present.

[0725] In some embodiments of the vaccine, the vaccine further comprises an amphiphile having the formula S-[B]-[U]-H,

[0726] Where S is the solubilization block;

[0727] B is a spacer;

[0728] H is a hydrophobic block;

[0729] U is a connector;

[0730] [ ] indicates that the group is optional;

[0731] – denotes that two adjacent groups are directly attached to each other by a covalent bond or indirectly attached to each other via a suitable linker X, and

[0732] wherein said amphiphile S comprises a dendrimer amplifier.

[0733] In some embodiments of the vaccine, S of the amphiphile comprises two or more solubilizing groups (SG) independently selected from amines, carboxylic acids or sugar molecules, wherein the sugar molecules are independently selected from mannose, sialyl Lewis x, sialyl Lewis a, Lewis y, Lewis x, Tn, sTn, TF, sTF, Globo H, SSEA-3, GM2, GD2, GD3 and fucosyl GM1 and combinations thereof.

[0734] In some embodiments of the vaccine, the at least one peptide antigen conjugate comprises at least one A selected from the group consisting of an autoantigen, an alloantigen, and an allergen.

[0735] In some embodiments of the vaccine, the vaccine further comprises at least one D selected from the group consisting of an inhibitor of mTOR, RORγt, CDK8 / 19, and HDAC, and an inhibitor of AHR, RAR, and A 2a agonists.

[0736] In some embodiments of the vaccine, the vaccine further comprises a second drug molecule (D2) independently selected from the group consisting of inhibitors of mTOR, RORγt, CDK8 / 19, and HDAC; inhibitors of AHR, RAR, and A 2a and an immunostimulatory agent selected from the group consisting of an agonist of NLR, CLR, TLR, and STING, with the proviso that D and D2 bind to different receptors.

[0737] In some embodiments of the vaccine, the D2 is selected from an agonist of an NLR, a CLR, a TLR, and STING. In other embodiments, the D2 is selected from an agonist of TLR-3, TLR-7, TLR-8, TLR-7 / 8, TLR-9, and STING. In other embodiments, the D2 is selected from an RNA and an imidazoquinoline agonist of TLR-7, TLR-8, and TLR-7 / 8.

[0738] In some embodiments of the vaccine, the vaccine further comprises a third drug molecule (D3) independently selected from the group consisting of inhibitors of mTOR, RORγt, CDK8 / 19 and HDAC; inhibitors of AHR, RAR and A 2a and an immunostimulatory agent selected from the group consisting of an agonist of NLR, CLR, TLR and STING, with the proviso that D, D2 and D3 bind to different receptors.

[0739] In some embodiments of the vaccine, the at least one D is selected from AZD-8055, AZD2016, KU-0063794, CC223, Torin-1, Torin-2, INK-128, WYE354, WYE132, OSI-027, OXA-01, PI-103, NVP-BEZ235, GNE-493, GSK2126458, rapamycin, tacrolimus, everolimus, RAD001, CCI-779 and AP23573.

[0740] In some embodiments of the vaccine, the molar ratio of total peptide antigen conjugate to at least one D is between about 20:1 and 1:2, or between about 10:1 and about 1:1, or between about 4:1 and about 2:1, preferably about 1:1.

[0741] In a preferred embodiment of the vaccine for inducing tolerance, the vaccine comprises at least one peptide antigen conjugate having a formula selected from the group consisting of PEG-[E1]-A-[E2]-[U]-H and H-[U]-[E1]-A-[E2]-PEG,

[0742] in

[0743] H is independently a hydrophobic block at each occurrence,

[0744] wherein one or more drug molecules (D) are optionally attached to each H directly or via a suitable linker X1;

[0745] A is independently a peptide antigen at each occurrence;

[0746] E1 is independently an N-terminal extension at each occurrence;

[0747] E2 is independently a C-terminal extension at each occurrence;

[0748] U is independently a linker at each occurrence;

[0749] wherein at least one A is selected from the group consisting of autoantigens, allergens, and alloantigens, and at least one D is present and selected from the group consisting of ATP-competitive mTOR inhibitors; [ ] indicates that the group is optional; and – indicates that two adjacent groups are directly attached to each other by a covalent bond or indirectly attached to each other via a suitable linker X.

[0750] In some embodiments of the vaccine, the at least one D is selected from AZD-8055, AZD2016, KU-0063794, CC223, Torin-1, Torin-2, INK-128, WYE354, WYE132, OSI-027, OXA-01, PI-103, NVP-BEZ235, GNE-493, GSK2126458, rapamycin, tacrolimus, everolimus, RAD001, CCI-779 and AP23573.

[0751] In some embodiments of the vaccine, the vaccine further comprises an amphiphile having the formula S-[B]-[U]-H,

[0752] Where S is the solubilization block;

[0753] B is a spacer;

[0754] H is a hydrophobic block;

[0755] U is a connector;

[0756] [ ] indicates that the group is optional; and

[0757] – means that two adjacent groups are directly attached to each other by a covalent bond or indirectly attached to each other via a suitable linker X,

[0758] wherein said amphiphile S comprises a dendrimer amplifier.

[0759] In some embodiments of the vaccine, the S of the amphiphile comprises two or more solubilizing groups (SG) independently selected from carboxylic acids, phosphoserines, and sugar molecules, wherein the sugar molecules are independently selected from mannose, glucose, glucosamine, N-acetylglucose, galactose, galactosamine, N-acetylgalactosamine, and agonists of CD22a.

[0760] In some embodiments of the vaccine, the peptide antigen conjugate has a net positive charge at physiological pH of between about +1 to about + 10. In other embodiments, the peptide antigen conjugate has a net positive charge at physiological pH of between about +2 to about +6 or between about +3 to about +5.

[0761] In some embodiments of the vaccine, the amphiphile is present and the molar ratio of peptide antigen conjugate to amphiphile is between about 4:1 and 1:20, preferably about 1:1.

[0762] In some embodiments of the vaccine, the amphiphile comprises a carboxylic acid and has a net negative charge. In other embodiments, the amphiphile comprises a carboxylic acid selected from beta alanine and succinic acid.

[0763] In some embodiments of the vaccine, the average net charge of the at least one peptide antigen conjugate is positive at physiological pH, and the molar ratio of the peptide antigen conjugate to the amphiphile is between about 4:1 and about 2:1, or between about 1:2 and about 1:16, or between about 1:2 and about 1:4. In certain preferred embodiments, the molar ratio is about 1:1.

[0764] In some embodiments of the vaccine, the vaccine comprises a particle further comprising an amphiphile and one or more peptide antigen conjugates. In a preferred embodiment of the vaccine, the vaccine comprises a particle comprising an amphiphile having the formula S-[B]-[U]-H and at least one peptide antigen conjugate having the formula PEG-[E1]-A-[E2]-[U]-H or H-[U]-[E1]-A-[E2]-PEG, wherein A is a peptide antigen, S is a solubilizing block; E1 and E2 are N-terminal extensions and C-terminal extensions, respectively; B is a spacer; U is a linker molecule; H is a hydrophobic block; [ ] indicates that a group is optional; – indicates that two adjacent groups are directly attached to each other by a covalent bond or indirectly attached to each other via a suitable linker X; and U and H of the amphiphile and the peptide antigen conjugate can be the same, different, or contain one or more identical functional groups or moieties.

[0765] In some embodiments of the vaccine, the amphiphile and / or peptide antigen conjugate further comprises one or more drug molecules (D). The drug molecule (D) can be directly linked to the hydrophobic block (H) of the amphiphile and / or peptide antigen conjugate or indirectly linked via X1 (e.g., S-[B]-[U]-HD and / or PEG-[E1]-A-[E2]-[U]-HD). The drug molecule (D) can be mixed with the amphiphile and / or peptide antigen conjugate (e.g., D + S-[B]-[U]-H + PEG-[E1]-A-[E2]-[U]-H), or the drug molecule (D) can be in the form of a drug molecule conjugate (i.e., D-[U]-H or HD), which is mixed with the amphiphile and / or peptide antigen conjugate (e.g., DH + S-[B]-[U]-H + PEG-[E1]-A-[E2]-[U]-H). Preferred compositions of vaccines further comprising a drug molecule (D) are described throughout this specification. Said D is directly bonded to the adjacent group or indirectly bonded to the adjacent group as a side chain or as part of a side chain group.

[0766] In a preferred embodiment of the vaccine, the vaccine comprises particles comprising an amphiphile and one or more peptide antigen conjugates, the vaccine further comprising a drug molecule (D) selected from immunomodulators. The drug molecule (D) selected from the group consisting of immunomodulators can be directly linked to the hydrophobic block (H) of the amphiphile and / or peptide antigen conjugate or indirectly linked via X1 (e.g., S-[B]-[U]-HD and / or PEG-[E1]-A-[E2]-[U]-HD); the drug molecule (D) can be mixed with the amphiphile and peptide antigen conjugate (e.g., D + S-[B]-[U]-H + PEG-[E1]-A-[E2]-[U]-H); or, the drug molecule (D) can be in the form of a drug molecule conjugate (i.e., D-[U]-H or HD) that is mixed with the amphiphile and peptide antigen conjugate (e.g., DH + S-[B]-[U]-H + PEG-[E1]-A-[E2]-[U]-H). Preferred compositions of vaccines further comprising a drug molecule (D) are described throughout this specification.

[0767] In some embodiments of vaccines for treating or preventing autoimmune diseases, the peptide antigen conjugate comprises an antigen (A) selected from autoantigens (sometimes referred to as self-antigens). In some embodiments of vaccines for treating or preventing allergies, the peptide antigen conjugate comprises an antigen (A) selected from allergens. In some embodiments of vaccines for treating or preventing transplant rejection, the peptide antigen conjugate comprises an antigen (A) selected from alloantigens. In some embodiments of vaccines for treating or preventing cancer, the peptide antigen conjugate comprises an antigen (A) selected from autoantigens, neoantigens or viral antigens. In some embodiments of vaccines for treating or preventing infectious diseases, the peptide antigen conjugate comprises an antigen (A) selected from viruses, bacteria, protozoa or fungi. Preferred antigens for treating different diseases and preferred methods for selecting antigens are described throughout this specification.

[0768] Particles comprising certain compositions of amphiphiles have been found to have particular utility for the delivery of small molecule drugs for a variety of applications, including the treatment of cancer, inflammation, autoimmune diseases, macular degeneration, and diseases of vital organs (including the liver) and metabolic disorders.

[0769] In some embodiments of the composition for cancer treatment, the cancer treatment comprises a particle comprising an amphiphile and a drug molecule selected from a chemotherapeutic agent and / or an immunomodulatory agent. In a preferred embodiment of the cancer treatment, the particle comprises an amphiphile and a drug D having the formula S-[B]-[U]-H, wherein S is a solubilizing block; B is a spacer; U is a linker molecule; H is a hydrophobic block; [ ] indicates that a group is optional; and the drug D is associated with the particle via covalent or non-covalent interactions.

[0770] In some embodiments, the drug molecule (D) is linked to the hydrophobic block (H) (where present) of the amphiphile, e.g., S-[B]-[U]-HD, wherein one or more Ds are directly bonded to an adjacent group, or indirectly bonded to an adjacent group at one or more termini via X1 or as part of a side chain group. In other embodiments, the drug molecule is mixed with the amphiphile (e.g., D + S-[B]-[U]-H) or linked to the hydrophobic block (H) and mixed with the amphiphile (e.g., D-[B]-[U]-H + S-[B]-[U]-H, or HD + S-[B]-[U]-H), and the drug is incorporated into particles formed from the amphiphile. Preferred compositions for cancer treatment comprising an amphiphile and at least one chemotherapeutic agent and / or immunostimulatory agent are described throughout this specification.

[0771] The present disclosure also relates to a peptide antigen conjugate having a formula selected from PEG-[E1]-A-[E2]-[U]-H-[D] and [D]-H-[U]-[E1]-A-[E2]-PEG, or a peptide antigen fragment having a formula selected from PEG-[E1]-A-[E2]-[U1] and [U1]-[E1]-A-[E2]-PEG.

[0772] The present disclosure also relates to a peptide antigen conjugate having a formula selected from PEG-[E1]-A-[E2]-[U]-H-[D] and [D]-H-[U]-[E1]-A-[E2]-PEG, or a peptide antigen fragment having a formula selected from PEG-[E1]-A-[E2]-[U1] and [U1]-[E1]-A-[E2]-PEG,

[0773] in

[0774] H is a hydrophobic block,

[0775] wherein one or more drug molecules (D) are optionally attached to each H directly or via a suitable linker X1;

[0776] A is a peptide antigen;

[0777] E1 is the N-terminal extension;

[0778] E2 is the C-terminal extension;

[0779] U is a connector;

[0780] U1 is the linker precursor;

[0781] [ ] indicates that the group is optional; and

[0782] – denotes that two adjacent groups are directly attached to each other by a covalent bond or indirectly attached to each other via a suitable linker X.

[0783] In some embodiments, the peptide antigen fragment has the formula A-[E2]-PEG.

[0784] In some embodiments, El and / or E2 are present and are selected from a cathepsin-cleavable tetrapeptide of the formula P4-P3-P2-P1.

[0785] In some embodiments, E1 and / or E2 are present and are selected from Ser-Pro-Val-Arg, Ser-Pro-Val-Cit, Val-Cit, and Ser-Pro-Val-aBut.

[0786] In some embodiments, the peptide antigen (A) comprises at least one amino acid selected from norleucine and alpha-aminobutyric acid.

[0787] In some embodiments, disclosed herein is a vaccine comprising a peptide antigen conjugate having a formula selected from the group consisting of PEG-[E1]-A-[E2]-[U]-H-[D] and [D]-H-[U]-[E1]-A-[E2]-PEG, or a peptide antigen fragment having a formula selected from the group consisting of PEG-[E1]-A-[E2]-[U1] and [U1]-[E1]-A-[E2]-PEG,

[0788] in

[0789] H is a hydrophobic block,

[0790] wherein one or more drug molecules (D) are optionally attached to each H directly or via a suitable linker X1;

[0791] A is a peptide antigen;

[0792] E1 is the N-terminal extension;

[0793] E2 is the C-terminal extension;

[0794] U is a connector;

[0795] U1 is the linker precursor;

[0796] [ ] indicates that the group is optional; and

[0797] – denotes that two adjacent groups are directly attached to each other by a covalent bond or indirectly attached to each other via a suitable linker X.

[0798] In some embodiments, vaccines used to induce tolerance can be used to prevent or treat autoimmune diseases. Non-limiting examples of autoimmune diseases include, but are not limited to, multiple sclerosis, anti-MOG, celiac disease (including refractory disease), type 1 diabetes, vitiligo, autoimmune hepatitis, neuromyelitis optica, autoimmune uveitis, rheumatoid arthritis, myasthenia gravis, Lambert-Eaton syndrome, Graves' disease, optic neuritis, immune thrombocytopenic purpura, pemphigus vulgaris, bullous pemphigoid, Goodpasture's syndrome, eczema, Sjögren's syndrome, achalasia, myositis, dermatomyositis, systemic sclerosis, psoriasis, inflammatory bowel disease (including Crohn's disease and ulcerative colitis), primary sclerosing cholangitis, various vasculitides (including but not limited to Takayasu's arteritis, giant cell arteritis, polyarteritis nodosa, Kawasaki disease, anti-GBM disease, ANCA vasculitis), systemic lupus erythematosus, amyotrophic lateral sclerosis (ALS), Behçet's disease, and birch pollen allergy.

[0799] The present disclosure also relates to a method of treating or preventing an autoimmune disease in a subject in need thereof, the method comprising administering to the subject a vaccine comprising at least one peptide antigen conjugate having a formula selected from the group consisting of PEG-[E1]-A-[E2]-[U]-H and H-[U]-[E1]-A-[E2]-PEG, and an amphiphile having the formula S-[B]-[U]-H,

[0800] in

[0801] A is a peptide antigen;

[0802] E1 is the N-terminal extension;

[0803] E2 is the C-terminal extension;

[0804] H is independently a hydrophobic block at each occurrence, wherein one or more drug molecules (D) are optionally attached to each H directly or via a suitable linker X1;

[0805] S is the solubilization block;

[0806] B is a spacer;

[0807] U is independently a linker at each occurrence;

[0808] [ ] indicates that the group is optional,

[0809] - means that two adjacent groups are directly attached to each other by a covalent bond or indirectly attached to each other via a suitable linker X;

[0810] wherein the amphiphile comprises a dendrimer amplifier and the at least one peptide antigen is selected from a self antigen or a tumor antigen.

[0811] In one embodiment of the method of treating an autoimmune disease, the vaccine is administered intravenously, subcutaneously, or intramuscularly.

[0812] The present disclosure also relates to a method for enhancing the efficacy and / or tolerability of a vaccine, the method comprising administering to the subject a vaccine comprising at least one peptide antigen conjugate having a formula selected from the group consisting of PEG-[E1]-A-[E2]-[U]-H and H-[U]-[E1]-A-[E2]-PEG, and an amphiphile having the formula S-[B]-[U]-H,

[0813] in

[0814] A is a peptide antigen;

[0815] E1 is the N-terminal extension;

[0816] E2 is the C-terminal extension;

[0817] H is independently a hydrophobic block at each occurrence, wherein one or more drug molecules (D) are optionally attached to each H directly or via a suitable linker X1;

[0818] S is the solubilization block;

[0819] B is a spacer;

[0820] U is independently a linker at each occurrence;

[0821] [ ] indicates that the group is optional, and

[0822] - means that two adjacent groups are directly attached to each other by a covalent bond or indirectly attached to each other via a suitable linker;

[0823] wherein the amphiphile comprises a dendrimer amplifier and at least one peptide antigen is selected from autoantigens.

[0824] Vaccines for inducing tolerance can be used to prevent or treat inflammatory diseases, which can be characterized as diseases in which an unwanted immune response is directed in a subject against an antigen, which can be a self-antigen, an alloantigen, a foreign antigen (e.g., an allergen), a drug molecule, or a device.

[0825] The present disclosure also relates to a method of inducing an immune response in a subject in need thereof, comprising administering to the subject at least one dose of a first vaccine (V1) followed by at least one dose of a second vaccine (V2), wherein V1 is a vaccine disclosed herein; and V2 is a viral vaccine.

[0826] In some embodiments, the subject's T cell response is increased relative to administration of only at least one dose of the first vaccine (V1).

[0827] In some embodiments, the subject's T cell response is increased relative to administration of only at least one dose of the second vaccine (V2).

[0828] In some embodiments, one dose of V1 is administered at a first time (V1T1). In other embodiments, two doses of V1 are administered at a first time (V1T1) and a second time (V1T2). In other embodiments, three doses of V1 are administered at a first time (V1T1), a second time (V1T2), and a third time (V1T3).

[0829] In some embodiments, one dose of V2 is administered at the first time (V2T1). In other embodiments, two doses of V2 are administered at the first time (V2T1) and the second time (V2T2). In other embodiments, three doses of V2 are administered at the first time (V2T1), the second time (V2T2), and the third time (V2T3).

[0830] In some embodiments, V1 is administered by intramuscular or intravenous route.

[0831] In some embodiments, V2 is administered intravenously.

[0832] In some embodiments, the initial dose of V2 is administered 1 to 6 weeks after the last dose of V 1. In other embodiments, the initial dose of V2 is administered 1 to 12 weeks after the last dose of V 1.

[0833] In some embodiments, V2 is an adenoviral vector vaccine.

[0834] In some embodiments, the adenovirus encodes a peptide antigen (A) of V1.

[0835] In some embodiments, V2 is a ChAdOx vaccine.

[0836] connector

[0837] The term linker refers to any molecule that joins any two or more molecules (or "parts") (such as any two or more components of an amphiphile, peptide antigen conjugate, or drug conjugate) together, and which can additionally perform any one or more of the following functions: I) increase or decrease water solubility; II) increase the distance between any two components; III) impart rigidity or flexibility; or, IV) regulate the degradation rate of the connection between any two or more different molecules. As used herein, the term "linker" can be used to describe a linker (U), a suitable linker (X) (such as X1, X2, X3, X4, and X5), and an extension (E1 or E2).

[0838] The linkers with specific utility are named, and the specific preferred compositions of those named linkers are described throughout this specification. Thus, extensions E1 and E2 are optional peptide-based linkers extending from the N-terminus and C-terminus of the peptide antigen (A), respectively, which can be included between the solubilizing block (S) (such as a PEG group or a charged block (C)) and the antigen (A), or between the antigen (A) and the hydrophobic block (H), or between the antigen (A) and the optional linker U. Spacer (B) is a linker between the solubilizing block (S) and the hydrophobic block (H) on the amphiphile. The molecule produced by the reaction of linker precursor 1 ("U1") (directly connected to the solubilizing block or drug (D) or indirectly connected via spacer (B)) and linker precursor 2 ("U2") on the hydrophobic block (H) is called linker U. A suitable linker X refers to any linker suitable for connecting two or more adjacent groups. A suitable linker preferably for joining a drug molecule (D) to a hydrophobic block (H) is called X1. Suitable linkers preferred for attaching aryl or heteroaryl groups to the hydrophobic block are designated as X2. Suitable linkers for attaching reactive functional groups ("FG4") to the pharmacophore of the drug molecule (D) are designated as X3. Suitable linkers preferred for attaching charged groups to the hydrophobic block (H) are designated as X4. Suitable linkers preferred for attaching SG to S are designated as X5.

[0839] The linker can use covalent or non-covalent means to join any two or more components. In a preferred embodiment, the linker can join (i.e., connect) any two components via a covalent bond. A covalent bond is a preferred connection for joining any two components and ensures that no component can be dispersed with other components immediately after administration to a subject.

[0840] There are many suitable linkers well known to those skilled in the art and include, but are not limited to, linear or branched carbon linkers, heterocyclic carbon linkers, rigid aromatic linkers, flexible ethylene oxide linkers, peptide linkers, or combinations thereof, which, for covalent linkers, further comprise two or more functional groups, which may be the same or different, for linking any two molecules (e.g., any two components of an amphiphile, peptide antigen conjugate, and / or drug conjugate) via a covalent bond.

[0841] In some embodiments, the carbon linker can include a C1-C18 alkane linker, for example, a lower alkyl linker, such as C1–C6 (i.e., one to six methylene units), which can be used to increase the spacing between two or more molecules (i.e., different components), while longer chain alkane linkers can be used to impart hydrophobic characteristics. Alternatively, a hydrophilic linker (such as an ethylene oxide linker) can be used instead of an alkane linker to increase the spacing between any two or more heterologous molecules and improve water solubility. In other embodiments, the linker can be a cyclic and / or aromatic compound or a poly(aromatic) compound that imparts rigidity. The linker molecule can comprise a hydrophilic or hydrophobic linker. In several embodiments, the linker includes a degradable peptide sequence that can be cleaved by an intracellular enzyme (such as a cathepsin or an immunoproteasome).

[0842] For linking two components of amphiphiles, peptide antigen conjugates, and drug conjugates, wherein at least one of the components comprises a peptide, it has been found that linkers comprising between 2 and 7 methylene groups improve the coupling of the two or more components. In a non-limiting example, increasing the number of methylene units between the amide and amine in the N-terminal amino acid of the peptide-based hydrophobic block (H) results in improved coupling to other molecules, including U2, antigen (A), extension E2, spacer (B), and solubilizing block (S) (such as PEG and charged block (C)). Thus, in a preferred embodiment, the N-terminal amino acid of the poly(amino acid)-based hydrophobic block (H) comprises two or more (typically between 2 and 7, such as 1, 2, 3, 4, 5, 6, 7) methylene units. For clarity, the amino acid having 2 methylene units is β-alanine, and the amino acid having 5 methylene units is amino-hexanoic acid. In certain preferred embodiments, the N-terminal amino acid of the peptide-based hydrophobic block (H) is amino-hexanoic acid (sometimes referred to as Ahx; CAS number 60-32-3). In other embodiments, the N-terminal amino acid of the peptide-based hydrophobic block (H) is beta-alanine.

[0843] In some embodiments, the joint can include poly (ethylene oxide) (PEG). The length of the joint depends on the purpose of the joint. For example, the length of the joint (such as PEG joint) can be increased to separate any two or more components, for example, to reduce steric hindrance, or in the case of a hydrophilic PEG joint, can be used to improve water solubility. The length of the joint such as PEG can be between about 1 and about 24 monomers, such as a length of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 monomers or longer. When used as spacer (B), the length of PEG can be up to 45 monomers or longer, but the length is usually between 4 and 36 monomers.

[0844] In some embodiments where the linker comprises a carbon chain, the linker can comprise a chain between about 1 or 2 and about 18 carbons in length, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18 carbons or longer. In some embodiments where the linker comprises a carbon chain, the linker can comprise a chain between about 12 and about 20 carbons. In some embodiments where the linker comprises a carbon chain, the linker can comprise a chain of no more than 18 carbons, typically between about 1 and 6 carbon atoms.

[0845] The linkage used to join any two or more molecules (e.g., any two or more components of an amphiphile, peptide antigen conjugate, and / or drug conjugate) can comprise any suitable functional group, including but not limited to amides, esters, ethers, thioethers, silyl ethers, disulfides, carbamates, carbonamides, hydrazides, hydrazones, acetals, and triazoles.

[0846] In a non-limiting example of covalent attachment, a click chemistry reaction can result in a triazole that connects (joins together) any two components of an amphiphile, a peptide antigen conjugate, or a drug molecule conjugate. In several embodiments, the click chemistry reaction is a strain-promoted [3+2] azide-alkyne cycloaddition reaction. The alkyne group and the azide group can be provided on the corresponding molecules to be connected by "click chemistry". In some embodiments, an antigen (A) carrying an azide functional group is coupled to a hydrophobic block (H) having a suitable reactive group (such as an alkyne, for example, dibenzylcyclooctyne (DBCO)).

[0847] In some embodiments, an amine is provided on one molecule and can be attached to another molecule by reacting the amine with any suitable electrophilic group such as a carboxylic acid, an acyl chloride, an activated ester (e.g., an NHS ester) to produce an amide bond; the amine can react with olefins (via Michael addition); the amine can react with aldehydes and ketones (via a Schiff base); or, the amine can react with an activated carbonate or carbamate to produce a carbamate.

[0848] In some embodiments, the linker is cleavable under intracellular conditions such that cleavage of the linker results in the release of any component attached to the linker, e.g., a drug molecule (D).

[0849] For example, the linker can be cleavable by an enzyme localized in an intracellular vesicle (e.g., a lysosome or endosome or caveolae) or an enzyme in the cytosol (e.g., a proteasome or immunoproteasome). The linker can be, for example, a peptide linker cleaved by a protease, including but not limited to a protease localized in an intracellular vesicle, such as a cathepsin in the lysosomal or endosomal compartment of the cell.

[0850] The length of the peptide linker is generally between 1-10 amino acids, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more (such as up to 20) amino acids in length, such as 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more amino acids in length. When used as a spacer (B), the peptide linker can be up to about 45 amino acids. It is known that some dipeptides are hydrolyzed by proteases, including cathepsins (such as cathepsins B and D) and plasmin (see, for example, Dubowchik and Walker, 1999, Pharm. Therapeutics 83: 67-123). For example, a peptide linker (e.g., Phe-Leu or Gly-Phe-Leu-Gly (SEQ ID NO: 1) linker) cleavable by the thiol-dependent protease cathepsin B can be used. Other examples of such linkers are described in, for example, U.S. Patent No. 6,214,345, incorporated herein by reference. In certain such embodiments, the peptide linker cleavable by an intracellular protease is a Val-Cit linker or a Phe-Lys linker (see, e.g., U.S. Patent No. 6,214,345, which describes the synthesis of doxorubicin with a Val-Cit linker). Note: For examples of amino acids and peptides provided throughout the specification (within the text of the figures), unless otherwise specified, it is understood that the peptides and amino acids are L-amino acids.

[0851] The cleavable peptide linker can be selected to promote processing (i.e., hydrolysis) of the peptide linker after intracellular uptake by immune cells. The sequence of the cleavable peptide linker can be selected to promote processing by intracellular proteases, such as cathepsins in intracellular vesicles or proteasomes or immunoproteasomes in the cytoplasmic cavity.

[0852] In some embodiments, recognition by cathepsins is facilitated using a linker comprising a peptide sequence of the formula Pn...P4-P3-P2-P1, wherein P1 is selected from arginine, lysine, acetyl lysine (i.e., the epsilon amine is acetylated), boc-protected lysine (i.e., the epsilon amine is boc-protected), citrulline, glutamine, threonine, leucine, norleucine, alpha-aminobutyric acid (abbreviated herein as "a-But"), or methionine; P2 is selected from glycine, serine, leucine, valine, or isoleucine; P3 is selected from glycine, serine, alanine, proline, or leucine; and P4 is selected from glycine, serine, arginine, lysine, acetyl lysine (i.e., the epsilon amine is acetylated), boc-protected lysine, aspartic acid, glutamic acid, or beta-alanine. In a non-limiting example, a tetrapeptide linker of the formula P4-P3-P2-P1 is linked to another molecule via an amide bond and has the sequence Lys-Pro-Leu-Arg (SEQ ID NO: 2). For clarity, the amino acid residues (Pn) are numbered from proximal to distal from the cleavage site, which is at the C-terminus of the P1 residue, e.g., the amide bond between P1-P1' is hydrolyzed. Suitable peptide sequences that promote cleavage by endosomal and lysosomal proteases such as cathepsins are well described in the literature (see: Choe et al., J. Biol. Chem., 281: 12824-12832, 2006).

[0853] In several embodiments, a linker comprising a peptide sequence is selected to promote recognition by the proteasome or immunoproteasome. The peptide sequence of the formula Pn...P4-P3-P2-P1 is selected to promote recognition by the proteasome or immunoproteasome, wherein P1 is selected from a basic residue and a hydrophobic branched residue, such as arginine, lysine, leucine, isoleucine, and valine; P2, P3, and P4 are optionally selected from leucine, isoleucine, valine, lysine, and tyrosine. In a non-limiting example, a cleavable linker of the formula P4-P3-P2-P1 recognized by the proteasome is linked to another molecule via an amide bond at P1 and has the sequence Tyr-Leu-Leu-Leu (SEQ ID NO: 3). Sequences that promote degradation by the proteasome or immunoproteasome can be used alone or in combination with a cathepsin cleavable linker. In some embodiments, amino acids that promote immunoproteasome processing are linked to a linker that promotes processing by endosomal proteases. A variety of suitable sequences that promote cleavage by the immunoproteasome are well described in the literature (see: Kloetzel et al., Nat. Rev. Mol. Cell Biol., 2:179-187, 2001; Huber et al., Cell, 148:727-738, 2012; and Harris et al., Chem. Biol., 8:1131-1141, 2001).

[0854] In certain preferred embodiments, the drug molecule (D) is linked to the hydrophobic block (H) via a linker X1 comprising an enzyme-degradable peptide. Non-limiting examples are shown here:

[0855]

[0856] wherein D is a drug molecule; "linker" is any suitable linker molecule; j represents any integer, however, j is typically 1 to 6 amino acids, such as 1, 2, 3, 4, 5 or 6 amino acids; R 8 is any suitable amino acid side group; the N-terminal amine of the peptide is directly linked to the N-terminus or C-terminus of, for example, a hydrophobic block (H) comprising a poly(amino acid) or is linked via a terminal link, either directly or via a U link, or via a reactive monomer comprising a hydrophobic block (H); and brackets "[ ]" indicate that a group is optional.

[0857] In certain preferred embodiments where the drug molecule is linked to the hydrophobic block (H) via a linker X1 comprising an enzymatically degradable peptide, the drug molecule (D) is directly linked to the peptide via an amide bond, as shown here:

[0858]

[0859] In the non-limiting example of the above structure, wherein the N-terminal linker group is present and is selected from beta-alanine, the structure is:

[0860]

[0861] In some embodiments, the drug molecule (D) is linked to the peptide via a suicide carbamate linker. Non-limiting examples are shown here:

[0862]

[0863] In the example above, where j is 4 and the amino acids are Ser-Lys(Ac)-Val-Norleucine, the structure is:

[0864]

[0865] In some embodiments, the drug molecule (D) is connected to the hydrophobic block (H) via a sulfatase degradable linker X1, wherein the hydrolysis of sulfate by the sulfatase results in the release of the drug molecule from the linker. A variety of arylsulfatase and alkylsulfatase degradable linkers have been recently described (e.g., see: Bargh et al., 2020, Chem. Sci. 11, 2375). In some embodiments of the present disclosure, the drug molecule is connected to the hydrophobic block (H) via a sulfatase degradable linker. For clarity, non-limiting examples are shown here:

[0866]

[0867] wherein D is a drug molecule; "linker" is any suitable linker molecule that is directly linked to the N-terminus or C-terminus of, for example, a hydrophobic block (H) comprising a poly(amino acid) or is linked via a terminal end, either directly or via a U, or via a reactive monomer comprising a hydrophobic block (H); and brackets "[ ]" indicate that a group is optional.

[0868] For clarity, non-limiting examples of the above structures are shown here, where a "linker" is present and is selected from a short alkyl linker attached to the hydrophobic block via an amide:

[0869] .

[0870] In other embodiments, any two or more components can be joined together via a pH-sensitive linker X that is sensitive to hydrolysis under acidic conditions. A variety of pH-sensitive linkers are well known to those skilled in the art and include, for example, hydrazones, carbonyl hydrazones, semicarbazones, thiosemicarbazones, cis-aconitamides, orthoesters, acetals, ketals, silyl ethers, and the like (see, e.g., U.S. Pat. Nos. 5,122,368, 5,824,805, 5,622,929; Dubowchik and Walker, 1999, Pharm. Therapeutics 83:67-123; Neville et al., 1989, Biol. Chem. 264:14653-14661).

[0871] In certain embodiments, the different components (e.g., a drug molecule and a hydrophobic block (H)) are linked together via a pH-sensitive linker that is stable at blood pH (e.g., at a pH of approximately 7.4) but undergoes more rapid hydrolysis at endosomal / lysosomal pH (approximately pH 5–6.5). In certain preferred embodiments, the drug molecule (D) is linked to the hydrophobic block (H) via a reactive monomer via a pH-sensitive bond (e.g., a hydrazone bond derived from the reaction between a ketone and a hydrazine). A functional group hydrazine linked to a carbonyl group is sometimes referred to as a hydrazide, however, hydrazine is intended to refer broadly to an -NH-NH2 group, including when linked to a carbonyl group, e.g., C(O)-NH-NH2. pH-sensitive linkages such as hydrazones offer the advantage that the bond is stable at physiological pH (approximately pH 7.4) but is hydrolyzed at lower pH values ​​(e.g., the pH of intracellular vesicles).

[0872] In certain preferred embodiments, the drug molecule is linked via a ketone-containing linker X1 and can be represented by the following formula:

[0873]

[0874] wherein D is any drug molecule; “linker” is any suitable linker molecule; y1 represents an integer between 1 and 6, preferably 4; brackets “[ ]” indicate that the group is optional; and wherein a ketone as in the above example is used to link the linker-attached drug molecule (D) to the reactive monomer via a hydrazone bond.

[0875] In the example above, where y1 is 4 and the drug molecule is directly attached via an amide bond (i.e., a "linker" is not present), the structure is:

[0876]

[0877] In a preferred embodiment, the drug molecule linked to the ketone is linked to the hydrophobic block (H) via a hydrazone or carbonyl hydrazone bond. Non-limiting examples of drug molecules linked to glutamic acid-based reactive monomers (N) via hydrazone and carbonyl hydrazone bonds are shown here:

[0878] 、 .

[0879] In some embodiments, the drug molecule comprises a ketone and can be directly linked to the reactive monomer via a hydrazone or carbonylhydrazone.

[0880] In other embodiments, the linker comprises a connection that can be cleaved under reducing conditions, such as a reducible disulfide bond. Many different linkers for introducing disulfide bonds are known in the art (see, for example, Thorpe et al., 1987, Cancer Res. 47: 5924-5931; Wawrzynczak et al., In Immunoconjugates: Antibody Conjugates in Radioimagery and Therapy of Cancer (CW Vogel, ed., Oxford U. Press, 1987); Phillips et al., Cancer Res. 68: 9280-9290, 2008). See also U.S. Patent No. 4,880,935.

[0881] In a preferred embodiment, the linker X1 connecting the hydrophobic block (H) to the one or more drug molecules (D) is a short alkyl or PEG linker. In other preferred embodiments, the linker X1 connecting the hydrophobic block (H) to the one or more drug molecules (D) is an enzyme-degradable linker, such as a cathepsin-degradable peptide or a sulfatase-degradable linker. In other preferred embodiments, the linker X1 connecting the hydrophobic block (H) to the one or more drug molecules (D) comprises an enzyme-degradable peptide and a suicide linker.

[0882] X can be any suitable linker, however, in preferred embodiments, the linker X connecting any two or more groups is a short alkyl group (i.e., a lower alkyl group) or a PEG linker, for example, a PEG linker having between about 1 and about 24 monomer units.

[0883] Extensions (E1 and E2)

[0884] The optional N-terminal extension and C-terminal extension (E1 and E2) represent moieties attached to the N-terminus and C-terminus of the peptide antigen (A), respectively. The N-terminal extension and C-terminal extension E1 and E2 may comprise any one or more of the following: amino acids, including non-natural amino acids; hydrophilic ethylene oxide monomers (e.g., PEG); hydrophobic alkane chains; or the like; or combinations thereof. The N-terminal extension and C-terminal extension E1 and E2 are attached to the peptide antigen (A) via any suitable means (e.g., via an amide bond).

[0885] In some embodiments, the extensions (E1 and E2) function to control the degradation rate of the peptide antigen (A), but may also perform any one or more additional functions. In some embodiments, the N-terminal or C-terminal extension (E1 or E2) can be free (wherein one end of the N-terminal or C-terminal extension is linked to the peptide antigen (A) and the other end is not linked to another molecule) and is used to slow the degradation of the peptide antigen; for example, the E1 peptide-based extension can be linked to the N-terminus of the peptide antigen via an amide bond to slow degradation. In other embodiments, the N-terminal extension and / or the C-terminal extension (E1 and / or E2) can be linked to a heterologous molecule and can function as a linker and be used to regulate the degradation of the peptide antigen (A). The N-terminal extension and / or the C-terminal extension that provides the linker function can directly link the peptide antigen to the hydrophobic block (H) and / or the solubilizing block (S) (such as PEG) or indirectly via a linker U. In some embodiments, the extension (E1 and / or E2) functions to provide distance (i.e., spacing) between any two heterologous molecules. In other embodiments, the extensions (E1 and / or E2) function to impart hydrophobic or hydrophilic properties to the peptide antigen conjugate. In still other embodiments, the composition of the extensions (E1 and / or E2) can be selected to impart rigidity or flexibility. In other embodiments, the N-terminal extension and / or the C-terminal extension (E1 and / or E2) can help stabilize the particles formed by the peptide antigen conjugate.

[0886] In some embodiments, the extension (E1 and / or E2) comprises a charged functional group that imparts a charge at pH 7.4, for example, a charged amino acid residue (e.g., arginine, ornithine, lysine, glutamic acid, aspartic acid, etc.). The number of charged residues present in the extension can be used to adjust the net charge of the peptide antigen conjugate. Peptide-based extensions (E1 and / or E2) that are recognized by proteases and impart a specific electrostatic charge to stabilize particles formed by the peptide antigen conjugate are described below.

[0887] In addition, in some embodiments, the C-terminal extension (E2) added to the peptide antigen (A) is selected to facilitate the production of a peptide comprising the formula PEG-[E1]-A-E2-[U1], wherein [ ] indicates that a group is optional. Thus, the amino acid sequence of the peptide-based E2 can be selected to disrupt peptide β-sheet formation and prevent sequence truncation during solid-phase peptide synthesis. In a non-limiting example, a C-terminal dipeptide linker (E2) Gly-Ser (e.g., Gly-Ser(Psi(Me,Me)pro)) is incorporated as a pseudo-proline dipeptide during solid-phase peptide synthesis. In another embodiment, proline is included in E2, e.g., Ser-Pro-Leu-Arg (SEQ ID NO: 4); thereby, the proline is included to both facilitate the production of the extension and promote processing of the extension by endosomal proteases.

[0888] In some embodiments, the peptide antigen (A) is linked to an E2 extension at the C-terminus, which is directly linked to the hydrophobic block or indirectly linked via a linker (U), for example, wherein the peptide antigen conjugate has the structure A-E2-UH or A-E2-H. In some embodiments, the E1 extension is linked to the N-terminus of the peptide antigen (A) and the E2 extension is linked to the C-terminus of the peptide antigen (A), wherein E1 or E2 is directly linked to the hydrophobic block (H) or linked via a linker (U), for example, wherein the peptide antigen conjugate has the structure E1-A-E2-UH, HU-E1-A-E2, E1-A-E2-H, or H-E1-A-E2. In other embodiments, the peptide antigen (A) is linked to an E1 extension at the N-terminus, which is directly linked to the hydrophobic block (H) or linked via a linker (U), for example, wherein the peptide antigen conjugate has the structure HU-E1-A or H-E1-A. In some embodiments, the solubilizing block, such as PEG, is linked to an extension E1 or E2, which is linked to the N-terminus or C-terminus of the peptide antigen (A), respectively, wherein the extension not linked to the solubilizing block (S), such as PEG, is directly linked to the hydrophobic block (H) or linked via a linker (U), for example, wherein the peptide antigen conjugate has the structure PEG-E1-A-E2-UH, HU-E1-A-E2-PEG, PEG-E1-A-E2-H or H-E1-A-E2-PEG.

[0889] In another embodiment, the PEG group is attached to both the E1 and E2 extensions, which are attached to both the N-terminus and C-terminus of the peptide antigen (A), respectively; or, PEG is attached to the E1 extension attached to the N-terminus of the peptide antigen (A) but not to the E2 extension attached to the C-terminus of the peptide antigen (A), which extensions can be directly attached to the hydrophobic block (H) or connected via a linker (U). The linker precursor U1 or the linker (U) can be attached to either extension (E1 or E2) via any suitable means, such as an amide bond.

[0890] In a preferred embodiment, the extensions (E1 and E2) are peptide sequences that are selected for recognition and hydrolysis by enzymes such as proteases. The extensions (E1 and E2) are preferably cleavable peptides that include amino acids recognized by either or both of endosomal proteases and / or immunoproteasomes.

[0891] In some embodiments, the N-terminal extension (E1) is a peptide sequence of about 1 to 8 amino acids in length (e.g., 1, 2, 3, 4, 5, 6, 7, or 8 amino acids, typically no more than 10 amino acids in length), which is linked to the peptide antigen (A) via an amide bond formed between the carboxyl group of E1 and the alpha amine of the N-terminal residue of the peptide antigen (A). The amide bond between E1 and the peptide antigen (A) can be cleaved by an enzyme.

[0892] The amino acid positions are generally numbered in order from proximal to distal from the cleavage site, with the amino acid position at the C-terminal end of the cleavage site indicated by a prime (e.g., Pn'). For example, for a tetrapeptide extension (PN4-PN3-PN2-PN1) linked to the N-terminus of a peptide antigen (A) that is an octapeptide (PA1'-PA2'-PA3'-PA4'-PA5'-PA6'-PA7'-PA8'), e.g., PN4-PN3-PN2-PN1-PA1'-PA2'-PA3'-PA4'-PA5'-PA6'-PA7'-PA8', the amide bond between PN1-PA1' is recognized and hydrolyzed by the enzyme.

[0893] In some embodiments, the N-terminal extension (E1) is an enzymatically degradable tetrapeptide recognized by endosomal proteases, wherein the PN1 position of the tetrapeptide extension (e.g., PN4-PN3-PN2-PN1) is preferably selected from arginine, lysine, citrulline, glutamine, threonine, leucine, norleucine or methionine, e.g., PN4-PN3-PN2-Arg; PN2 is selected from glycine, valine, leucine or isoleucine; PN3 is selected from glycine, serine, alanine, proline or leucine; and PN4 is selected from glycine, serine, arginine, lysine, aspartic acid or glutamic acid. In some embodiments, the N-terminal extension (E1) is an enzymatically degradable tripeptide recognized by an endosomal protease, wherein the PN1 position of the tripeptide extension (e.g., PN3-PN2-PN1) is preferably selected from arginine, lysine, citrulline, glutamine, threonine, leucine, norleucine, or methionine; PN2 is selected from glycine, valine, leucine, or isoleucine; and PN3 is selected from glycine, serine, alanine, proline, or leucine. In some embodiments, the N-terminal extension (E1) is an enzymatically degradable dipeptide recognized by an endosomal protease, wherein the PN1 position of the dipeptide extension (e.g., PN2-PN1) is preferably selected from arginine, lysine, citrulline, glutamine, threonine, leucine, norleucine, or methionine; and PN2 is selected from glycine, valine, leucine, or isoleucine. In still other embodiments, the N-terminal extension (E1) is an amino acid recognized by endosomal proteases, wherein the N1 position is preferably selected from arginine, lysine, citrulline, glutamine, threonine, leucine, norleucine or methionine. In a preferred embodiment of a vaccine for inducing tolerance, the E1 comprising the dipeptide is valine-citrulline.

[0894] In other embodiments, the N-terminal extension (E1) is an enzymatically degradable peptide recognized by the immunoproteasome, wherein the P1 position of the tetrapeptide extension (PN4-PN3-PN2-PN1) is preferably selected from isoleucine, leucine, norleucine or valine, for example, PN4-PN3-PN2-Leu.

[0895] In another embodiment, the N-terminal extension (E1) is an enzymatically degradable peptide recognized by both endosomal proteases and immunoproteasomes, wherein the PN5 and PN1 positions of the octapeptide extension (PN8-PN7-PN6-PN5-PN4-PN3-PN2-PN1) are selected from arginine, lysine, citrulline, glutamine, threonine, leucine, norleucine or methionine (for the PN5 position recognized by cathepsins) and isoleucine, leucine, norleucine or valine (for the PN1 position recognized by immunoproteasomes); for example, PN8-PN7-PN6-Arg-PN4-PN3-PN2-Leu. A non-limiting example of an N-terminal extension (E1) recognized by both cathepsins and immunoproteasomes is Lys-Pro-Leu-Arg-Tyr-Leu-Leu-Leu (SEQ ID NO: 5).

[0896] Non-limiting examples of tetrapeptide N-terminal extensions (E1) recognized by the immunoproteasome include: Ser-Leu-Val-Cit (SEQ ID NO:6), Ser-Leu-Val-Leu (SEQ ID NO:7), Ser-Pro-Val-Cit (SEQ ID NO:8), Glu-Leu-Val-Arg (SEQ ID NO:9), Ser-Pro-Val-Arg (SEQ ID NO:10), Ser-Leu-Val-Arg (SEQ ID NO:11), Lys-Pro-Leu-Arg (SEQ ID NO:2), Lys-Pro-Val-Arg (SEQ ID NO:12), Glu-Leu-Val-Cit (SEQ ID NO:13), Glu-Leu-Val-Leu (SEQ ID NO:14), Glu-Pro-Val-Cit (SEQ ID NO:15), and Lys-Pro-Val-Cit (SEQ ID NO:16). Non-limiting examples of tripeptide N-terminal extensions (E1) include: Leu-Val-Cit, Leu-Val-Leu, Pro-Val-Cit, Leu-Val-Arg, Pro-Val-Arg, Pro-Leu-Arg, Gly-Val-Ser. Non-limiting examples of dipeptide N-terminal extensions (E1) include: Val-Cit, Val-Leu, Val-Arg, Leu-Arg. Non-limiting examples of single amino acid N-terminal extensions (E1) include Cit, Arg, Leu, or Lys. In the above examples, Arg can be replaced by Lys; Lys can be replaced by Arg; Glu can be replaced by Asp; and Asp can be replaced by Glu. Note that Cit = citrulline.

[0897] In some embodiments, E2 is a degradable peptide connected to the C-terminal residue of the peptide antigen (A) and comprises an amino acid sequence that is recognized and hydrolyzed by certain proteases. In some embodiments, the C-terminal extension (E2) is a peptide sequence having a length of about 1 to 8 amino acids (such as 1, 2, 3, 4, 5, 6, 7 or 8 amino acids, usually not more than 10 amino acids). In a preferred embodiment, the C-terminal extension (E2) is connected to the peptide antigen (A) via an amide bond formed between the C-terminal carboxyl group of the peptide antigen (A) and the alpha amine of the N-terminal residue of the extension (E2). The amide bond between E2 and the peptide antigen (A) can be cleaved by an enzyme. Note: Amino acid positions are usually numbered in order from the proximal to the distal end of the cleavage site, where the amino acid position at the C-terminal end of the cleavage site is represented by a prime symbol (e.g., Pn'). For example, for a tetrapeptide extension (PC1′-PC2′-PC3′-PC4′) linked to the C-terminus of an octapeptide antigen (PA8-PA7-PA6-PA5-PA4-PA3-PA2-PA1), e.g., PA8-PA7-PA6-PA5-PA4-PA3-PA2-PA1-PC1′-PC2′-PC3′-PC4′, the amide bond between PA1-PC1′ is recognized and hydrolyzed by the enzyme.

[0898] In a preferred embodiment of the C-terminal extension (E2), the C-terminal extension (E2) comprises an amino acid sequence selected to promote recognition and cleavage by immunoproteasomes and, optionally, recognition by endosomal proteases. Because peptide antigen (A) typically contains a C-terminal residue (e.g., leucine) that promotes hydrolysis by immunoproteasomes (e.g., at an amide bond near the C-terminal residue of the peptide antigen (A)), the extension attached to the C-terminus of the peptide antigen (A) should be selected to promote recognition and cleavage by immunoproteasomes at an amide bond near the C-terminus of the peptide antigen (A). Immunoproteasomes prefer small, uncharged amino acids at the PC1' position adjacent to the C-terminal amino acid PA1 of the peptide antigen (A), for example, the amide bond between PA1-PC1'. However, endosomal proteases prefer bulky, hydrophobic amino acids (e.g., leucine, norleucine, methionine, or glutamine) and basic amino acids (i.e., arginine and lysine). Therefore, the C-terminal extension can be selected to promote recognition by either or both classes of proteases.

[0899] In some embodiments, a peptide antigen (A) having the sequence PA8-PA7-PA6-PA5-PA4-PA3-PA2-PA1 is linked to a C-terminal peptide extension (E2) having the sequence PC1' ... PCn', where n is an integer value from 1 to 8, e.g., PA8-PA7-PA6-PA4-PA3-PA2-PA1-PC1' ... PCn'. The composition of the C-terminal extension (E2) depends on the length of the extension sequence used. In some embodiments, the C-terminal extension E2 is a single amino acid PC1' selected from Gly, Ala, Ser, Arg, Lys, Cit, Gln, Thr, Leu, Nle, or Met. In other embodiments, the C-terminal extension E2 is a dipeptide PC1'-PC2', where PC1' is selected from Gly, Ala, or Ser; and PC2' is selected from Gly, Ala, Ser, Pro, Arg, Lys, Cit, Gln, Thr, Leu, Nle, or Met. In a further embodiment, the C-terminal extension E2 is a tripeptide PC1′-PC2′-PC3′, wherein P1′ is selected from Gly, Ala or Ser; PC2′ is selected from Gly, Ala, Ser or Pro; and PC3′ is selected from Gly, Ser, Arg, Lys, Cit, Gln, Thr, Leu, Nle or Met.

[0900] In another embodiment, the C-terminal extension E2 is a tetrapeptide extension PC1'-PC2'-PC3'-PC4', wherein PC1' is selected from glycine, alanine or serine; PC2' is selected from glycine, alanine, serine, proline or leucine; PC3' is selected from glycine, alanine, serine, valine, leucine or isoleucine; and PC4' is selected from arginine, lysine, citrulline, glutamine, threonine, leucine, norleucine or methionine. In another embodiment, the C-terminal extension E2 is a pentapeptide PC1'-PC2'-PC3'-PC4'-PC5', wherein PC1' is selected from glycine, alanine or serine; PC2' is selected from glycine, alanine, serine, proline, arginine, lysine, glutamic acid or aspartic acid; PC3' is selected from glycine, alanine, serine, proline or leucine; PC4' is selected from glycine, alanine, valine, leucine or isoleucine; and PC5' is selected from arginine, lysine, citrulline, glutamine, threonine, leucine, norleucine or methionine. In another embodiment, the C-terminal extension E2 is a hexapeptide PC1'-PC2'-PC3'-PC4'-PC5'-PC6', wherein PC1' is selected from glycine, alanine or serine; PC2' is selected from glycine, alanine, serine or proline; PC3' is selected from glycine, serine, proline, arginine, lysine, glutamic acid or aspartic acid; PC4' is selected from proline or leucine; PC5' is selected from glycine, alanine, valine, leucine or isoleucine; and PC6' is selected from arginine, lysine, citrulline, glutamine, threonine, leucine, norleucine or methionine.

[0901] Non-limiting examples of hexapeptide C-terminal extensions (E2) include Gly-Gly-Lys-Leu-Val-Arg (SEQ ID NO: 17), Gly-Gly-Lys-Pro-Leu-Arg (SEQ ID NO: 18), Gly-Gly-Ser-Leu-Val-Arg (SEQ ID NO: 19), Gly-Gly-Ser-Leu-Val-Cit (SEQ ID NO: 20), Gly-Gly-Ser-Pro-Val-Cit (SEQ ID NO: 21), Gly-Gly-Ser-Leu-Val-Leu (SEQ ID NO: 22), Gly-Gly-Glu-Leu-Val-Arg (SEQ ID NO: 23), and Gly-Gly-Glu-Leu-Val-Leu (SEQ ID NO: 24).

[0902] Non-limiting examples of pentapeptide C-terminal extensions (E2) include Gly-Ser-Leu-Val-Arg (SEQ ID NO: 25), Gly-Ser-Leu-Val-Cit (SEQ ID NO: 26), Gly-Lys-Pro-Val-Cit (SEQ ID NO: 27), Gly-Lys-Pro-Val-Arg (SEQ ID NO: 28), Gly-Ser-Leu-Val-Leu (SEQ ID NO: 29), Gly-Glu-Leu-Val-Leu (SEQ ID NO: 30).

[0903] Non-limiting examples of tetrapeptide C-terminal extensions (E2) include Ser-Leu-Val-Cit (SEQ ID NO: 6), Ser-Leu-Val-Leu (SEQ ID NO: 7), Ser-Pro-Val-Cit (SEQ ID NO: 8), Glu-Leu-Val-Arg (SEQ ID NO: 9), Ser-Pro-Val-Arg (SEQ ID NO: 10), Ser-Leu-Val-Arg (SEQ ID NO: 11), Lys-Pro-Leu-Arg (SEQ ID NO: 2), Glu-Leu-Val-Cit (SEQ ID NO: 13), Glu-Leu-Val-Leu (SEQ ID NO: 14), Glu-Pro-Val-Cit (SEQ ID NO: 15), Glu-Gly-Val-Cit (SEQ ID NO: 31).

[0904] Non-limiting examples of tripeptide C-terminal extensions (E2) include Gly-Ser-Gly, Gly-Ser-Arg, Gly-Ser-Leu, Gly-Ser-Cit, Gly-Pro-Gly, Gly-Pro-Arg, Gly-Pro-Leu, Gly-Pro-Cit. Non-limiting examples of dipeptide C-terminal extensions (E2) include Gly-Ser, Gly-Pro, Val-Cit, Gly-Arg, Gly-Cit. Non-limiting examples of single amino acid C-terminal extensions (E2) include Gly, Ser, Ala, Arg, Lys, Cit, Val, Leu, Met, Thr, Gln, or Nle. In the above examples, Arg can be replaced by Lys; Lys can be replaced by Arg; Glu can be replaced by Asp; and Asp can be replaced by Glu.

[0905] The C-terminal linker (E2) attached to the C-terminus of the peptide antigen (A) can be selected for recognition (i.e., hydrolysis) by both immunoproteasomes and endosomal proteases. In a non-limiting example, a peptide antigen (A) having the sequence PA8-PA7-PA6-PA5-PA4-PA3-PA2-PA1 is linked at the C-terminus to a C-terminal tetrapeptide extension (E2) having the sequence PC1'-PC2'-PC3'-PC4', wherein PC1' is selected from glycine, alanine, or serine and PC4' is selected from arginine, lysine, citrulline, glutamine, threonine, leucine, norleucine, or methionine, e.g., Ser-P3-P2-Arg. In some embodiments, an antigen having the sequence PA8-PA7-PA6-PA5-PA4-PA3-PA2-PA1 is linked at the C-terminus to a C-terminal hexapeptide extension (E2) having the sequence PC1'-PC2'-PC3'-PC4'-PC5'-PC6', wherein PC1' and PC2' are selected from glycine, alanine, proline, or serine, and PC6' is selected from arginine, lysine, citrulline, glutamine, threonine, leucine, norleucine, or methionine, e.g., Gly-Gly-PC3'-PC4'-PC5'-Arg. A non-limiting example of a C-terminal extension (E2) linked to the C-terminus of the peptide antigen (A) that promotes processing by both the immunoproteasome and cathepsins is Gly-Gly-Lys-Pro-Leu-Arg (SEQ ID NO: 18). Additional non-limiting examples of C-terminal extensions (E2) attached at the C-terminus of the peptide antigen (A) that facilitate processing by immunoproteasomes and cathepsins are Gly-Gly-Ser-Leu-Val-Cit (SEQ ID NO: 20) or Gly-Gly-Ser-Pro-Val-Cit (SEQ ID NO: 21).

[0906] Spacer (B)

[0907] The spacer (B) is an optional component of the amphiphile that connects the solubilizing block (S) to the hydrophobic block (H) directly or via a linker (U), for example, where the amphiphile has the structure SBH or SBUH. The spacer (B) may comprise any one or more of the following: amino acids, including unnatural amino acids; hydrophilic polymers, for example, polymers based on: monomers based on ethylene oxide (PEG), acrylates, methacrylates, acrylamides, or methacrylamides; alkane chains; and the like; or combinations thereof. The spacer (B) may be linked to the solubilizing block (S) and the hydrophobic block (H) by any suitable means, for example, directly or indirectly via a linker, but the linkage typically comprises a covalent bond, for example, an amide bond.

[0908] In some embodiments, the spacer (B) functions to provide distance (i.e., spacing) between the heterologous molecules S and H. In other embodiments, the spacer (B) functions to impart hydrophobic or hydrophilic properties. In still other embodiments, the composition of the spacer can be selected to impart rigidity or flexibility. In other embodiments, the composition of the spacer can be selected for recognition by an enzyme and promote degradation.

[0909] In some embodiments, the spacer (B) is a hydrophilic polymer whose monomer units are selected from acrylates, (meth)acrylates, acrylamides, (meth)acrylamides, allyl ethers, vinyl acetate, vinylamides, substituted styrenes, amino acids, acrylonitrile, heterocyclic monomers (e.g., ethylene oxide), sugars, phosphates, phosphamides, sulfonates, sulfonamides, or combinations thereof.

[0910] In some embodiments, the spacer (B) is a peptide sequence of about 1 to 45 amino acids in length (such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 29, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44 or 45 amino acids, typically not more than 45 amino acids in length), which is connected to the hydrophobic block (H) and the solubilizing block (S), respectively, via, for example, an amide bond formed between the N-terminal carboxyl group and the C-terminal carboxyl group of the spacer (B). The amide bond between the spacer (B) and the solubilizing block (S) and / or the hydrophobic block (H) may be recognized by an enzyme or may be selected to resist enzyme-mediated hydrolysis.

[0911] In other embodiments, the spacer (B) is a hydrophilic polymer comprising monomeric units selected from non-natural hydrophilic monomers (e.g., ethylene oxide (PEG), HPMA, poly(sarcosine) or HEMA), the length of the hydrophilic polymer (i.e., the degree of polymerization) is about 1 to 48 monomers, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 29, 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 or 48 monomers, typically not more than 48 monomers in length, and the spacer is directly connected to the hydrophobic block (H) and the solubilizing block (S) or connected via a linker.

[0912] The specific composition of the spacer that leads to the unexpected improvement in biological activity is described throughout this specification. Note: The spacer group (B) and the solubilizing block (S) can both comprise a hydrophilic polymer (e.g., a hydrophilic poly(amino acid); a hydrophilic methacrylate-based polymer such as HEMA; a hydrophilic methacrylamide-based polymer such as HPMA, PEG, etc.); however, the distinction between S and B is based in part on function and is noted in the specific example of amphiphiles. Similarly, the PEG group of the peptide antigen conjugates of the formula PEG-[E1]-A-[E2]-[U]-H and H-[E1]-A-[E2]-[U]-PEG is a hydrophilic polymer and a type of solubilizing block.

[0913] Connector (U)

[0914] The linker (U) joins the solubilizing block (S) fragment (S-[B]-U1) to the hydrophobic block (H) fragment (U2-H), optionally via reaction of U1 with U2, to form an amphiphile (S-[B]-UH).

[0915] The linker (U) also joins the peptide antigen conjugate fragment (PEG-[E1]-A-[E2]-U1 or U1-[E1]-A-[E2]-PEG) to the hydrophobic block (H) fragment (U2-H) independently of the amphiphilic linker U via the reaction of U1 and U2 to form a peptide antigen conjugate (PEG-[E1]-A-[E2]-UH or HU-[E1]-A-[E2]-PEG).

[0916] Although the peptide antigen (A) can be conjugated to the hydrophobic block (H) entirely on the resin by solid phase peptide synthesis, either directly (i.e., AH) or via an extension (i.e., A-E2-H or H-E1-A), in some cases it may be beneficial to produce the antigen (A) and the hydrophobic block (H) as separate fragments comprising a linker precursor U1 (PEG-[E1]-A-[E2]-U1 or U1-[E1]-A-[E2]-PEG) and a linker precursor U2 (U2-H), which can be conjugated on the resin or in solution to produce PEG-[E1]-A-[E2]-UH (or HU-[E1]-A-[E2]-PEG).

[0917] Similarly, although the solubilizing block (S) on the amphiphile can be joined to the hydrophobic block (H) directly (i.e., SH) or via a spacer (i.e., SBH) entirely on the resin by solid phase peptide synthesis, in some cases it may be beneficial to produce the solubilizing block (S) and the hydrophobic block (H) as separate fragments comprising a linker precursor U1 (S-[B]-U1) and a linker precursor U2 (U2-H), which can be joined on the resin or in solution to produce S-[B]-UH.

[0918] In preferred embodiments, the linker precursor used to form linker U is selected for site selectivity, i.e., the reaction occurs only between U1 and U2 and not between other groups. In some embodiments, linker precursor U1 comprises an activated carboxylic acid and reacts with linker precursor U2 comprising an amine to form a linker U comprising an amide; alternatively, U1 comprises an amine and reacts with U2 comprising an activated carboxylic acid to form a linker U comprising an amide. In some embodiments, linker precursor U1 comprises a maleimide and reacts with linker precursor U2 comprising a thiol to form a linker U comprising a thioether bond; alternatively, U1 comprises a thiol and reacts with U2 comprising a maleimide to form a linker U comprising a thioether bond. In some embodiments, linker precursor U1 comprises an azide and reacts with linker precursor U2 comprising an alkyne to form a linker U comprising a triazole; alternatively, U1 comprises an alkyne and reacts with U2 comprising an azide to form a linker US comprising a triazole.

[0919] In a preferred embodiment, an amphiphile of formula S-[B]-UH is joined together by linking a solubilizing block segment (S-[B]-U1) to a hydrophobic block segment (U2-H), wherein the linker precursor U1 comprises a strained alkyne (e.g., dibenzocyclooctyne (DBCO), bicyclononyne (BCN), etc.), which reacts with an azide-containing linker precursor U2 to form a triazole-containing linker U.

[0920] In a preferred embodiment, the peptide antigen conjugate of the formula PEG-[E1]-A-[E2]-UH or HU-[E1]-A-[E2]-PEG is joined together by linking the peptide antigen fragment PEG-[E1]-A-[E2]-U1 or U1-[E1]-A-[E2]-PEG to the hydrophobic block fragment (U2-H), wherein the linker precursor U1 comprises a strained alkyne (e.g., dibenzocyclooctyne (DBCO), bicyclononyne (BCN), etc.), which reacts with the linker precursor U2 comprising an azide to form a linker U comprising a triazole.

[0921] In a preferred method of manufacture, a peptide antigen conjugate of the formula PEG-[E1]-A-[E2]-UH or HU-[E1]-A-[E2]-PEG is joined together by linking the peptide antigen fragment PEG-[E1]-A-[E2]-U1 or U1-[E1]-A-[E2]-PEG to a hydrophobic block fragment (U2-H), wherein the linker precursor U1 comprises DBCO and the linker precursor U2 comprises an azide, by: (i) adding 1 molar equivalent of the peptide antigen fragment at a concentration greater than 10 mM, preferably greater than 25 mM in DMSO to at least 1.05 equivalents of the hydrophobic block fragment at a concentration greater than 25 mM, most preferably greater than 50 mM in DMSO; and (ii) after the reaction is complete, removing any unreacted hydrophobic block fragment by adding an azide-resin (such as agarose-azide) to the reaction mixture; and then (iii) removing the resin to produce the pure peptide antigen conjugate.

[0922] In other preferred embodiments, linker precursor U1 comprises an azide that reacts with a linker precursor U2 comprising a strained alkyne (eg, dibenzocyclooctyne (DBCO), bicyclononyne (BCN), etc.) to form a linker U comprising a triazole. In a non-limiting example, a DBCO-containing linker precursor U2 is linked to the hydrophobic block (H) via a suitable linker X (e.g., DBCO-NHS, CAS No. 1353016-71-3), and a linker precursor U1 (e.g., an azido acid, such as azidopentanoic acid; an azido amino acid, such as azido-lysine (abbreviated as Lys(N3)), CAS No. 159610-92-1; or an azido amine, such as azido-butylamine) is linked to the solubilizing block fragment (S-[B]-U1) or the peptide antigen fragment (PEG-[E1]-A-[E2]-U1 or U1-[E1]-A-[E2]-PEG) via a suitable linker X.

[0923] In a preferred embodiment, the linker U preferably comprises an amide, a thioether or a triazole.

[0924] Dendrimer Amplifier

[0925] Dendrimer amplifiers are a specific class of linker moieties that function to increase the valence (i.e., number) of groups present on any component of the amphiphile, peptide antigen conjugate, or drug molecule conjugate described herein. For example, in a preferred embodiment of the solubilizing block (S), a dendrimer amplifier is used to increase the valence of the solubilizing groups (referred to as "SG" in the formula) present on the surface of the solubilizing block (S). In other embodiments, a dendrimer amplifier is used to increase the valence of the solubilizing block (S) and spacer (B) attached to the hydrophobic block (H).

[0926] Dendrimer amplifiers (also known as "dendrimers") are regularly branched molecules that are generally symmetrical and typically comprise repeating units of monomers containing three or more functional groups (FG) and branching points. Dendrimer amplifiers can be represented by the formula (FG')-T-(FGt)d, where FG' and FGt are the sink and terminal functional groups, respectively, selected from any suitable functional groups; T is any suitable linker, and "d" is any integer greater than 1, typically between 2 and 32, but more preferably between 2 and 8, such as 2, 3, 4, 5, 6, 7, and 8. The multiple by which a dendrimer amplifier increases the terminal functional group (FGt) can be represented by FGt = β γ , where β is the number of branches present per generation of dendrons, and the symbol γ is the number of generations, wherein the number of branches is any integer, but is typically between 2 and 6, and the number of generations is any integer, but is typically between 1 and 10. The free (i.e., unreacted) terminal functional groups present on the solubilizing block may also be referred to as solubilizing groups (SG).

[0927] A dendrimer amplifier can comprise repetitions of a monomer comprising a first functional group (FG1) and a second functional group (FG2), wherein the first functional group is reactive toward the second functional group. For example, for clarity, a non-limiting example of a second generation dendrimer amplifier with β = 2 is shown here, comprising repetitions of a monomer comprising a first functional group (FG1) and a second functional group (FG2), wherein the first functional group is reactive toward the second functional group:

[0928]

[0929] Among them, the first functional group at the starting point is also called the sink functional group (FG'), and the end FG2 is called the terminal functional group or FGt.

[0930] For clarity, a non-limiting example of a 3rd generation dendron formed from monomers comprising first and second functional groups is shown here, where β = 2:

[0931]

[0932] For clarity, a non-limiting example of a 2nd generation dendrimer amplifier with β = 3 comprising repeats of a first monomer comprising a first functional group (FG1) and a second functional group (FG2), wherein the first functional group is reactive toward the second functional group, is shown here:

[0933]

[0934] The monomer comprising a first functional group and a second functional group can be selected from any suitable monomer, wherein the first functional group is reactive toward the second functional group and the monomer comprises at least one first functional group and two or more second functional groups. Non-limiting examples include FG1-(CH2) y2 CH(R 1 )2、FG1-(CH2) y2 C(R 1 )3、FG1-(CH2CH2O) y2 CH(R 1 )2、FG1-(CH2CH2O) y2 C(R 1 )3、FG1-CH(R 1 )2、FG1-C(R 1 )3, where R 1 Independently selected from (CH2) y3 -FG2, (OCH2CH2) y3 -FG2 or CH2(OCH2CH2) y3 -FG2), and y2 and y3 are each an integer number selected from between 1 and 6 repeating units.

[0935] For clarity, FG1-CH(R 1 )2 is shown here as a non-limiting example, wherein FG1 is NH2, R 1 is CH2(OCH2CH2) y3 -FG2, y3 is 1 and FG2 is COOH:

[0936]

[0937] The above monomers were used to generate the second generation amplification linker, the structure of which is:

[0938]

[0939] Additional non-limiting examples of monomers comprising a first functional group and a second functional group, wherein the first functional group is reactive toward the second functional group, and the monomer comprises at least one first functional group and two or more second functional groups, include FG1-(CH2) y2 N(R 2 )2、FG1-(CH2CH2O) y2 CH2CH2N(R 2 )2, where R 2 Independently selected from (CH2) y3 -FG2, (CH2CH2O) y3 (CH2) y4 -FG2, (CH2OCH2CH2)y3 -FG2), and y2, y3 and y4 are each an integer selected from repeating units between 1 and 6. Note: In the above examples, FG' is an amine and the four FGt are carboxylic acids.

[0940] For clarity, FG1-(CH2CH2O) y1 CH2CH2N(R 2 )2 is shown here as a non-limiting example, wherein FG1 is NH2, R 2 is (CH2CH2O) y3 (CH2) y4 -FG2, y2 is 2, y3 is 1, y4 is 2 and FG2 is COOH:

[0941]

[0942] Still further non-limiting examples of monomers comprising a first functional group and a second functional group, wherein the first functional group is reactive toward the second functional group, and the monomer comprises at least one first functional group and two or more second functional groups, include certain amino acids, such as glutamic acid, aspartic acid, lysine, or ornithine. For clarity, non-limiting examples of 3rd generation lysine dendrimers are shown here:

[0943]

[0944] A dendrimer amplifier can comprise repeats of two monomers, wherein the first monomer comprises three or more first functional groups (FG1) and the second monomer comprises two or more second functional groups (FG2), wherein the first functional group is reactive toward the second functional group. For example, for clarity, a non-limiting example of a second generation dendrimer amplifier with β = 2 is shown here, comprising repeats of first and second monomers, wherein the first monomer comprises three first functional groups (FG1) and the second monomer comprises two second functional groups (FG2), wherein the first functional group is reactive toward the second functional group:

[0945]

[0946] For clarity, a non-limiting example of a 1st generation dendrimer amplifier with β = 2, comprising repeats of first and second monomers, wherein the first monomer comprises three first functional groups (FG1) and the second monomer comprises three second functional groups (FG2), wherein the first functional groups are reactive toward the second functional groups, is shown here:

[0947]

[0948] Any three or more components of the amphiphile, peptide antigen conjugate, and drug molecule conjugate can be joined together using dendrimer amplifiers. The sink functional group (FG') and the terminal functional group (FGt) can be further functionalized, ie, reacted to suit a specific purpose.

[0949] In a preferred embodiment of the amphiphile of formula S-[B]-[U]-H, the solubilizing block (S) comprises a dendrimer amplifier, wherein the sink is directly linked to the hydrophobic block (H) or indirectly linked via a spacer (B) and / or a linker U, and the terminal functional group (FGt) is unattached and serves as a solubilizing group or is linked to a solubilizing group (SG). The solubilizing group (SG) is any hydrophilic and / or charged molecule; preferably, the solubilizing group (SG) is described throughout this specification.

[0950] In some embodiments of the amphiphile of formula S-[B]-[U]-HD, the peptide antigen conjugate of formula PEG-[E1]-A-[E2]-[U]-HD or HD-[U]-[E1]-A-[E2]-PEG, and the drug molecule conjugate of formula HD, the hydrophobic block (H) comprises a dendrimer amplifier, wherein the sink is (i) directly connected to the solubilizing block (S) or indirectly connected via a spacer (B) and / or a linker U, (ii) directly connected to the antigen (A) or indirectly connected via an extension (E1 or E2) and / or a linker U; or (iii) directly connected to the drug molecule or connected via a linker X1.

[0951] In some embodiments, the hydrophobic block (H) comprises a dendrimer amplifier and a terminal functional group (FGt) is linked to a hydrophobic drug molecule. In such embodiments, the sink (i) is directly linked to the solubilizing block (S) or indirectly linked via a spacer (B) and / or a linker U, (ii) is directly linked to the antigen (A) or indirectly linked via an extension (E1 or E2) and / or a linker U; or (iii) is unreacted or capped with a terminal group such as an acetyl group. Capping or capping refers to modifying a functional group such as FGt to reduce its reactivity and / or to have a neutral charge at pH 7.4. For example, an amine can be capped with an activated carboxylic acid (e.g., acetyl chloride) to obtain a relatively less reactive amide; or, for example, a strained alkyne can be capped with an alkyl-azide to obtain a relatively less reactive triazole.

[0952] Hydrophobic block (H)

[0953] The hydrophobic block (sometimes referred to as "H" in the formula) is a molecule with significantly limited water solubility, or has amphiphilic properties, and is capable of assembling into supramolecular structures (e.g., micelles, nanoparticles, or microparticles) in aqueous solution. In certain embodiments, the hydrophobic block (H) is insoluble or forms micelles in aqueous solution at a concentration of less than about 1.0 mg / mL (e.g., about 0.1 mg / mL or about 0.01 mg / mL). In some embodiments, the hydrophobic block is soluble in aqueous solution within certain concentration, temperature, and / or pH ranges, but becomes insoluble in response to changes in concentration, temperature, and / or pH. For example, in some embodiments, the hydrophobic block is a temperature-responsive hydrophobic polymer, i.e., a hydrophobic polymer that is soluble in aqueous solution at a temperature below a transition temperature (T tr ), but becomes insoluble at temperatures above the transition temperature. Preferably, the hydrophobic block (H) is a molecule having a solubility of at least less than about 1.0 mg / mL, such as less than about 0.1 mg / mL or less than about 0.01 mg / mL, at or near physiological pH (about pH 7.4), between about pH 6.5 and pH 8.5, or between about pH 6.0 and pH 9.0, and at or near physiological temperature (about 37° C.), and at or near physiological salt concentration (about 10 g / L) and salt composition.

[0954] The hydrophobic block (H) may be selected from any molecule comprising higher alkanes, cyclic aromatic compounds, fatty acids, terpene / isoprene derived compounds, or polymers or oligomers with limited water solubility and / or amphiphilic character.

[0955] Exemplary higher alkanes include, but are not limited to, octane, nonane, decane, undecane, dodecane, tridecane, tetradecane, pentadecane, hexadecane, heptadecane, and octadecane. Exemplary cyclic aromatic compounds include, but are not limited to, phenyl. Exemplary saturated and unsaturated fatty acids include, but are not limited to, myristic acid, palmitic acid, stearic acid, or oleic acid. In some embodiments, the hydrophobic block (H) is a fatty acid, such as myristic acid. In other embodiments, the hydrophobic block (H) comprises a diacyl lipid, such as 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine or 1,2-distearoyl-sn-glycero-3-phosphoethanolamine; or a lipopeptide, such as Pam2Cys. In some embodiments, the fatty acid or lipid-based hydrophobic block (H) may further comprise PEG. Exemplary terpene / isoprene-derived compounds include sterol derivatives, such as cholesterol and squalene. In some embodiments, the hydrophobic block (H) comprises cholesterol. In some embodiments, the hydrophobic block (H) comprises a saponin, eg, QS-21.

[0956] In some embodiments, the hydrophobic block (H) is a linear, branched, or brush polymer (or oligomer). The hydrophobic block (H) may be a homopolymer or a copolymer. The hydrophobic block (H) may comprise one or more different types of monomer units. The hydrophobic block (H) may be a statistical copolymer or an alternating copolymer. The hydrophobic block (H) may be a block copolymer, such as an AB type, or the polymer may comprise a graft copolymer, whereby two or more polymers are linked by a polymer-analogous reaction.

[0957] The hydrophobic block (H) may comprise a polymer containing naturally occurring and / or non-natural monomers and combinations thereof.

[0958] In some embodiments, the hydrophobic block (H) is selected from natural biopolymers. Natural biopolymers can include peptides (sometimes referred to as poly (amino acids)) comprising hydrophobic amino acids. Non-limiting examples of hydrophobic amino acids include leucine, isoleucine, norleucine, valine, tryptophan, aniline, tyrosine and methionine, as well as hydrophilic amino acids that have been modified (such as by acetylation or benzoylation) to have hydrophobic characteristics. Natural biopolymers that are water-soluble in their natural form can be used, but they must be chemically modified to make such natural biopolymers water-insoluble and suitable for use as hydrophobic blocks (H). For example, biopolymers comprising hydrophilic amino acids such as glutamic acid or lysine residues can be modified at the γ carboxyl group or the ε amine group, respectively, for attaching hydrophobic molecules such as hydrophobic drug molecules to increase the hydrophobicity of the resulting modified biopolymer. Similarly, the biopolymer may be selected from hydrophilic polysaccharides, which may include but are not limited to glycogen, cellulose, dextran, alginate, and chitosan, but such polysaccharides should be chemically modified, for example by acetylation or benzoylation of the hydrophilic functional groups, to render the resulting modified polysaccharide water-insoluble. In still other embodiments, the hydrophobic block comprises monomers selected from lactic acid and / or glycolic acid.

[0959] The monomers constituting the hydrophobic block (H) may be selected from acrylates, (meth)acrylates, acrylamides, (meth)acrylamides, allyl ethers, vinyl acetate, vinylamides, substituted styrenes, amino acids, acrylonitriles, heterocyclic monomers (e.g., ethylene oxide), sugars, phosphates, phosphoramides, sulfonates, sulfonamides, or combinations thereof. Specific examples of (meth)acrylates and (meth)acrylamides include benzyl methacrylamide (BnMAM) and benzyl methacrylate (BnMA), respectively.

[0960] Certain monomers described herein as hydrophobic monomers may be water-soluble in aqueous solution under certain conditions, but hydrophobic and water-insoluble under certain conditions. Non-limiting examples include temperature-responsive monomers such as N-isopropylmethacrylamide (NIPMAM); a homopolymer composed entirely of NIPMAM may be water-soluble at room temperature, but may become insoluble and form particles at elevated temperatures. Such distinctions are made to facilitate the description of certain embodiments. In some embodiments, the hydrophobic block comprises a majority of monomer units selected from hydrophobic monomers having temperature responsiveness (sometimes referred to as "temperature responsive monomers"), such as NIPAM, NIPMAM, N,N'-diethylacrylamide (DEAAM), N-(L)-(1-hydroxymethyl)propylmethacrylamide (HMPMAM), N,N'-dimethylaminoethyl methacrylate (DMEMA), N-(N-ethylcarbonamido)propylmethacrylamide, N-vinylisobutyramide (PNVIBA), N-vinyl-n-butyramide (PNVBA), N-acryloyl-N-propylpiperazine (PNANPP), N-vinylcaprolactam (PVCa), DEGMA, TEGMA, or poly(amino acid) or γ-(2-methoxyethoxy)ester-L-glutamate. In still other embodiments, the hydrophobic block (H) may comprise monomers of ethylene oxide, propylene oxide, or a combination thereof.

[0961] The hydrophobic block (H) comprising a polymer generally comprises a hydrophobic monomer and one or more other types of monomers, such as reactive monomers, spacer monomers and / or charged monomers optionally linked to a drug molecule. In some embodiments of the hydrophobic block (H) comprising a polymer (or oligomer), the majority of monomer units are selected from hydrophobic monomers. In other embodiments of the hydrophobic block (H) comprising a polymer (or oligomer), the majority of monomer units are selected from reactive monomers linked to a hydrophobic drug molecule. In still other embodiments of the hydrophobic block (H) comprising a polymer (or oligomer), the polymer comprises a hydrophobic monomer and a reactive monomer linked to a hydrophobic drug molecule. In still other embodiments of the hydrophobic block (H) comprising a polymer (or oligomer), the polymer comprises a hydrophobic monomer and a charged monomer and a reactive monomer optionally linked to a hydrophobic drug molecule.

[0962] In preferred embodiments, the hydrophobic block (H) comprises a polymer (or oligomer) comprising a hydrophobic monomer further comprising an aromatic group. In certain embodiments, the hydrophobic block (H) comprises a heteroaryl group. In still other embodiments, the aryl or heteroaryl group of the hydrophobic block (H) comprises an amino substituent. The present inventors have discovered that hydrophobic blocks (H) comprising aminoaryl or aminoheteroaryl groups result in improved manufacturability and solubility in water-miscible solvents. The present inventors have also discovered that amphiphiles having a hydrophobic block (H) comprising an aromatic amine result in the formation of stable particles with a low CMC.

[0963] In a preferred embodiment, the hydrophobic block (H) comprises a monomer containing an aryl or heteroaryl group. Exemplary aryl groups (sometimes referred to as "aromatic compounds" or "aromatic rings") include, but are not limited to, phenyl, naphthyl, and quinolyl. Non-limiting examples include:

[0964]

[0965]

[0966] , where X is any suitable linker molecule and y is an integer value, typically between 1 and 6.

[0967] In preferred embodiments, aryl or heteroaryl groups include but are not limited to

[0968] 、 、 and .

[0969] In addition, one or more hydrogen atoms in the aforementioned aryl or heteroaryl groups may be replaced with one or more fluorine atoms. In certain embodiments, the hydrophobic block comprises a fluorinated aliphatic, aryl, or heteroaryl group, wherein one or more hydrogen atoms of the aforementioned group comprising the hydrophobic monomer may be replaced with one or more fluorine atoms. The following non-limiting examples of fluorinated aryl groups may be present in the hydrophobic monomer:

[0970]

[0971] , where X is any suitable linker molecule and y is an integer value, typically between 1 and 6.

[0972] The present inventors have surprisingly discovered that a hydrophobic block (H) comprising an aminoaryl or aminoheteroaryl group results in improved manufacturing and solubility in polar aprotic solvents and alcohols. Thus, in certain preferred embodiments, the hydrophobic block (H) comprises a moiety of the formula -Ar-NHR, wherein Ar can be aryl or heteroaryl, and R is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, or heteroarylalkyl. Non-limiting examples of aminoaryl or aminoheteroaryl groups include, but are not limited to:

[0973] and , where X is any suitable linker molecule and y is an integer value, typically between 1 and 6.

[0974] In some embodiments, the hydrophobic block (H) comprises a polymer (or oligomer) that further comprises a hydrophobic monomer having a fused aromatic group (e.g., naphthyl) or a fused heteroaromatic group (e.g., xanthene or quinolinyl). In some embodiments, the hydrophobic block (H) comprises a reactive monomer linked to a hydrophobic drug molecule. In some embodiments, the hydrophobic drug molecule (e.g., imidazoquinoline) is aromatic, and thus the reactive monomer linked to the hydrophobic drug molecule containing an aromatic group can also be described as a hydrophobic monomer containing an aromatic group or a reactive monomer linked to the drug.

[0975] In some embodiments, the hydrophobic block (H) comprises a poly(amino acid) of Formula I:

[0976]

[0977] wherein the poly(amino acid) of formula I comprises monomers selected from the group consisting of hydrophobic amino acids (M), reactive amino acids (N), spacer amino acids (O), charged amino acids (P), and combinations thereof, with the proviso that at least monomer M or N is present; m, n, o, and p represent integers of repeating units of monomers M, N, O, and P, respectively, which may be distributed along the polymer in a specific or random order; and R3 is typically selected from hydrogen, NH2, NH2-CH3, NH2-(CH2), y5 CH3, OH or a drug molecule (D) connected directly or via X1.

[0978] In some embodiments, P is absent. In other embodiments, N, O, and P are each absent.

[0979] In some embodiments, P is , where each R 5 Independently, a group comprising 1 to 2 charged functional groups.

[0980] In some embodiments, O is , wherein each Q is independently selected from (CH2) y6 and (CH2CH2O) y7 CH2CH2; each y6 is independently selected from an integer from 1 to 6; and each y7 is independently selected from an integer from 1 to 4.

[0981] In some embodiments, N is , wherein each X1 is independently a suitable linker; and each D is independently a drug molecule.

[0982] In some embodiments, M is , where each R 4 are independently hydrophobic groups.

[0983] In some embodiments, the hydrophobic block (H) comprises a poly(amino acid) of Formula I:

[0984]

[0985] wherein the poly(amino acid) of formula I comprises monomers selected from the group consisting of a hydrophobic amino acid (M), a reactive amino acid (N), a spacer amino acid (O), a charged amino acid (P), and combinations thereof, with the proviso that at least monomer M or N is present; m, n, o, and p represent integers of repeating units of monomers M, N, O, and P, respectively, which monomers may be distributed along the polymer in a specific or random order; R 3 Typically selected from hydrogen, NH2, NH2-CH3, NH2-(CH2) y5 CH3, OH or a drug molecule (D) directly connected or connected via X1; R 4 is any hydrophobic group typically selected from aryl or heteroaryl; R 5 is any group containing one or more functional groups that are charged in aqueous solution or are pH-responsive and charged in aqueous solution within a certain pH range; Q is typically selected from any lower alkyl or heteroalkyl group, including but not limited to (CH2) y6 and (CH2CH2O) y7 CH2CH2, wherein y6 is any integer from 1 to 6, and y7 is an integer generally selected from 1 to 4; and the N-terminus is (i) directly linked to the solubilizing block (S) or indirectly linked via a spacer (B) and / or a linker U; (ii) directly linked to the peptide antigen (A) or indirectly linked via an extension (E1 or E2) and / or a linker U; or (iii) directly linked to the drug molecule or linked via X1. Note: Hydrophobic amino acids, reactive amino acids, spacer amino acids, and charged amino acids are sometimes more generally described as hydrophobic monomers, reactive monomers, spacer monomers, and charged monomers, respectively.

[0986] In a preferred embodiment of the poly(amino acid) of formula I, R 4 yes

[0987]

[0988] in,

[0989] α is aryl or heteroaryl;

[0990] X2 is present or absent and, when present, is a suitable linker;

[0991] y8 is an integer selected from 0 and 6; and

[0992] Z 1 、Z 2 and Z 3 Each is independently selected from H, F, hydroxy, amino, alkyl and fluoroalkyl.

[0993] In preferred embodiments of the poly(amino acids) of formula I, α is aryl, for example, phenyl or naphthyl. In other embodiments, α is heteroaryl, for example, imidazolyl, pyridyl, quinolinyl, isoquinolinyl, indolyl, and benzimidazolyl.

[0994] In preferred embodiments of the poly(amino acid) of formula I, X2 is absent. In other embodiments, X2 is present and is selected from C(O), CO2(CH2) y9 , and C(O)NH(CH2) y9 , NHC(O) and NHC(O)(CH2) y9 , wherein y9 is an integer generally selected from 1 to 6. In other embodiments, X2 is present and is selected from lower alkyl and PEG groups.

[0995] In a preferred embodiment of the poly(amino acid) of formula I, the poly(amino acid) of formula I comprises a hydrophobic amino acid M selected from the group consisting of 4 In a preferred embodiment, R 4 is selected from the group consisting of hydrophobic groups comprising aryl, heteroaryl, aminoaryl and / or aminoheteroaryl groups. 4 Non-limiting examples include, but are not limited to:

[0996] 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 and , wherein X2 is any suitable linker molecule, and y8 is an integer value, typically between 0 and 6. In a preferred embodiment, y8 is 1.

[0997] In a non-limiting example, where R 4 yes , monomer M is:

[0998] .

[0999] In some embodiments, the poly(amino acid)-based hydrophobic block (H) of Formula I comprises a reactive amino acid N selected from any natural or unnatural amino acid, wherein the drug molecule (D) is linked to the monomer directly or via X. Suitable reactive amino acids include, but are not limited to, any amino acid carrying a group suitable for drug molecule attachment, including amino acids with the following groups: azide, alkyne, tetrazine, trans-cyclooctyne (TCO), protected hydrazine, ketone, aldehyde, certain hydroxyl, isocyanate, isothiocyanate, carboxylic acid, activated carboxylic acid, activated carbamate, activated carbamate, protected maleimide, thiol and / or amine groups.

[1000] X1 is any suitable linker for connecting the drug molecule D to the hydrophobic block (H) (including connecting to the reactive amino acid N of the poly(amino acid)), and is typically selected from -(CH2) y10 -FG3 and -(CH2) y10 -R 6 (or -C(O)-(CH2) y10 -FG3 and -C(O)-(CH2) y10 -R 6 , when the drug is attached at the N-terminus or outside the amine group, or -NH-(CH2) y10 -FG3 and -NH-(CH2) y10 -R 6 , when the drug is attached at the C-terminus or outside the carbonyl group), wherein y10 is any integer, generally selected from 1 to 6, and R 6 Typically selected from any one or more of the following: -C(O)-NH-R 7 、-NH-C(O)-R 7 、-NH-C(O)-OR 7 、-OC(O)-NH-R 7 、-OC(O)-R 7 、-C(O)-OR 7 、-OR 7 , OC(O)-W or -C(O)-W, where R 7 Typically selected from any one or more of the following: -(CH2) y11 -W, -(CH2) y11 -(OCH2CH2) y12 -W, -(CH2) y11 -(OCH2CH2) y12 -(CH2) y13 -W, –CHR8 -C(O)-W、–CHR 8 -C(O)-(NH-CHR 8 -C(O)) j -W、–(CH2) y11 -C(O)-NH-CHR 8 -C(O)-W、–(CH2) y11 -C(O)-NH-CHR 8 -C(O)-(NH-CHR 8 -C(O)) j -W、–(CH2) y11 -(OCH2CH2) y12 -C(O)-NH-CHR 8 -C(O)-W、–(CH2) y11 -(OCH2CH2) y12 -(CH2) y13 C(O)-NH-CHR 8 -C(O)-W、–(CH2) y11 -(OCH2CH2) y12 -C(O)-NH-CHR 8 -C(O)-(NH-CHR 8 -C(O)) j -W、–(CH2) y11 -(OCH2CH2) y12 -(CH2) y13 -C(O)-NH-CHR 8 -C(O)-(NH-CHR 8 -C(O)) j -W、–CHR 8 -C(O)-NH-C6H4-CH2-O-C(O)-W、–CHR 8 -C(O)-NH(CH3)(CH2)2-O-C(O)-W、–CHR 8 -C(O)-(NH-CHR 8 -C(O)) j -NH-C6H4-CH2-O-C(O)-W、–CHR 8 -C(O)-(NH-CHR 8 -C(O)) j -NH(CH3)(CH2)2-O-C(O)-W、–(CH2) y11 -C(O)-(NH-CHR 8 -C(O)) j -NH-C6H4-CH2-O-C(O)-W、–(CH2)y11 -C(O)-(NH-CHR 8 -C(O)) j -NH(CH3)(CH2)2-O-C(O)-W、–(CH2) y11 -(OCH2CH2) y12 -C(O)-(NH-CHR 8 -C(O)) j -NH-C6H4-CH2-O-C(O)-W、–(CH2) y11 -(OCH2CH2) y12 -C(O)-(NH-CHR 8 -C(O)) j -NH(CH3)(CH2)2-O-C(O)-W、–(CH2) y11 -(OCH2CH2) y12 -(CH2) y13 C(O)-(NH-CHR 8 -C(O)) j -NH-C6H4-CH2-O-C(O)-W、–(CH2) y11 -(OCH2CH2) y12 -(CH2) y13 C(O)-(NH-CHR 8 -C(O)) j -NH(CH3)(CH2)2-O-C(O)-W、–(CH2) y11 -(OCH2CH2) y12 -(CH2) y13 -C(O)-NH-(CH2) y14 -C(O)-(NH-CHR 8 -C(O)) j -NH-C6H4-CH2-O-C(O)-W、–(CH2) y11 -(OCH2CH2) y12 -(CH2) y13 C(O)-NH-(CH2) y14 -C(O)-(NH-CHR 8 -C(O)) j -NH(CH3)(CH2)2-O-C(O)-W、–(CH2) y11 -(OCH2CH2) y12 -C(O)-NH-(CH2) y14 -C(O)-(NH-CHR 8 -C(O)) j-NH-C6H4-CH2-OC(O)-W、–(CH2) y11 -(OCH2CH2) y12 -C(O)-NH-(CH2) y14 -C(O)-(NH-CHR 8 -C(O)) j -NH(CH3)(CH2)2-OC(O)-W、-CHR 8 -C(O)-NH-(CH2) y15 -W, -CHR 8 -NH-C(O)-(CH2) y15 -W, -CHR 8 -C(O)-(NH-CHR 8 -C(O)) j -NH-(CH2) y15 -W, -CHR 8 -NH-(C(O)-CHR 8 -NH) j -C(O)-(CH2) y15 -W, wherein y11, y12, y13, y14, y15 and j are each independently selected from any integer generally selected from 1 to 6, R 8 is any amino acid side group, and W can be independently selected from H (hydrogen), FG3, LG, and w; wherein FG3 is any suitable functional group for attachment to a drug molecule, which can be selected from, but is not limited to, carboxylic acids, activated carboxylic acids (e.g., carbonyl thiazolidine-2-thione ("TT"), NHS, or nitrophenol esters), carboxylic anhydrides, amines, and protected amines (e.g., tert-butoxycarbonyl protected amines), OSi(CH3), alkenes, azides, alkynes, strained alkynes, halogens (e.g., fluorine, chlorine), alkenes and endocyclic alkenes (e.g., allyl), CN, OH, and epoxy groups, hydrazines (including hydrazides), carbohydrazides, aldehydes, ketones, carbamates, and activated carbamates. , LG is any suitable leaving group, which can be selected from but not limited to any suitable leaving group (e.g., NHS, TT, nitrophenol, etc.), and w is a group derived from the reaction of FG4 with FG3 or the displacement of LG by FG4, and is typically selected from NH-, C(O)-, NH-C(O)-, C(O)-NH-, OC(O)-NH-, C(O)-NH-N=C(CH3)-, NH-N=C(CH3)- or -C(CH3)=N-NH-C(O)-, wherein w is always directly connected to D (i.e., wD) or indirectly connected via X3 (i.e., w-X3-D).

[1001] The drug molecule (D) can be attached to the reactive amino acid N directly or via X1 by reacting FG4 with FG3, wherein FG4 is any suitable functional group on the drug (D) that can react with FG3. Alternatively, the drug molecule (D) can be linked to the reactive amino acid N via X1 by replacing LG with any suitable FG4 containing a nucleophile (e.g., a primary amine), or the drug molecule (D) can be linked to the reactive amino acid N via X1 by replacing LG present on the drug molecule with any suitable FG3 containing a nucleophile.

[1002] In a preferred embodiment, FG3 is a carboxylic acid and FG4 is an amine, which react to form an amide. In a non-limiting example, X1 is selected from -(CH2) y10 -FG3, y10 is 2, FG3 is a carboxylic acid, and FG4 present on the drug is an amine (i.e., NH2-D), which react to form an amide, which can be represented as -(CH2)2-C(O)-D (amine not shown) or -(CH2)2-C(O)-NH-D (amine shown), thereby indicating that the drug is attached via an amide bond at the carbonyl group of X1, which (after amide bond formation) can be described as -(CH2) y10 -R 6 , where y10 is 2, R 6 = C(O)-W, and W is a group w that is NH- and is linked to D to give -(CH2)2-C(O)-NH-D.

[1003] The drug may further comprise a linker X3 between the reactive functional group FG4 and the pharmacophore, for example, FG4-X3-D. Specific preferred compositions of X3 are described elsewhere.

[1004] In other embodiments, FG3 is an amine and FG4 is a carboxylic acid, which react to form an amide. In a non-limiting example, X1 is -(CH2) y10 -FG3, y10 is 4, FG3 is an amine, and FG4 present on the drug is a carboxylic acid (i.e., COOH-D), which react to form an amide, which can be represented as -(CH2)4-NH-D (carbonyl not shown) or -(CH2)4-NH-C(...

Claims

1. A vaccine comprising at least one peptide antigen conjugate having a formula selected from the group consisting of PEG-[E1]-A-[E2]-[U]-H and H-[U]-[E1]-A-[E2]-PEG, wherein A is a peptide antigen; E1 is the N-terminal extension; E2 is the C-terminal extension; H is independently a hydrophobic block at each occurrence, wherein one or more drug molecules (D) are optionally attached to each H directly or via a suitable linker X1; U is independently a linker at each occurrence; [ ] indicates that the group is optional, and - means that two adjacent groups are directly attached to each other by a covalent bond or indirectly attached to each other via a suitable linker X.

2. The vaccine of claim 1 , wherein the vaccine further comprises an amphiphile having the formula S-[B]-[U]-H, wherein S is a solubilizing block; B is a spacer; H is a hydrophobic block; U is a connector; [ ] indicates that the group is optional; and – means that two adjacent groups are directly attached to each other by a covalent bond or indirectly attached to each other via a suitable linker X, wherein said S of said amphiphile comprises a dendron amplifier.

3. The vaccine of claim 1, wherein the PEG group of the peptide antigen conjugate comprises between 12 and 36 monomer units.

4. The vaccine of claim 1 , wherein E1 is present and comprises an enzymatically degradable peptide sequence comprising: i) a single amino acid PN1, wherein the PN1 is selected from arginine, lysine, citrulline, glutamine, threonine, leucine, norleucine and methionine; ii) a dipeptide PN2-PN1, wherein PN1 is selected from arginine, lysine, citrulline, glutamine, threonine, leucine, norleucine and methionine, and PN2 is selected from glycine, valine, leucine and isoleucine; iii) a tripeptide PN3-PN2-PN1, wherein PN1 is selected from arginine, lysine, citrulline, glutamine, threonine, leucine, norleucine, and methionine; PN2 is selected from glycine, valine, leucine, and isoleucine; and PN3 is selected from glycine, serine, alanine, proline, and leucine; or iv) the tetrapeptide PN4-PN3-PN2-PN1, wherein PN1 is selected from arginine, lysine, citrulline, glutamine, threonine, leucine, norleucine and methionine; PN2 is selected from glycine, valine, leucine and isoleucine; PN3 is selected from glycine, serine, alanine, proline and leucine; and PN4 is selected from glycine, serine, arginine, lysine, aspartic acid and glutamic acid.

5. The vaccine of claim 1 , wherein E2 is present and comprises an enzyme-degradable peptide sequence comprising: i) a single amino acid PC1′, wherein the PC1′ is selected from the group consisting of glycine, serine, arginine, lysine, citrulline, glutamine, threonine, leucine, norleucine, and methionine; ii) a dipeptide PC1′-PC2′, wherein PC1′ is selected from glycine and serine; and PC2′ is selected from glycine, serine, proline, arginine, lysine, citrulline, glutamine, threonine, leucine, norleucine, and methionine; iii) a tripeptide PC1′-PC2′-PC3′, wherein PC1′ is selected from glycine and serine; PC2′ is selected from glycine, serine, and proline; and PC3′ is selected from glycine, serine, arginine, lysine, citrulline, glutamine, threonine, leucine, norleucine, and methionine; iv) a tetrapeptide PC1′-PC2′-PC3′-PC4′, wherein PC1′ is selected from glycine and serine; PC2′ is selected from glycine, serine, proline, and leucine; PC3′ is selected from glycine, valine, leucine, and isoleucine; and PC4′ is selected from arginine, lysine, citrulline, glutamine, threonine, leucine, norleucine, and methionine; v) a pentapeptide PC1′-PC2′-PC3′-PC4′-PC5′, wherein PC1′ is selected from glycine and serine; PC2′ is selected from glycine, serine, proline, arginine, lysine, glutamic acid, and aspartic acid; PC3′ is selected from glycine, serine, proline, and leucine; PC4′ is selected from glycine, valine, leucine, and isoleucine; and PC5′ is selected from arginine, lysine, citrulline, glutamine, threonine, leucine, norleucine, and methionine; or vi) the hexapeptide PC1′-PC2′-PC3′-PC4′-PC5′-PC6′, wherein PC1′ is selected from glycine and serine; PC2′ is selected from glycine, serine and proline; PC3′ is selected from glycine, serine, proline, arginine, lysine, glutamic acid and aspartic acid; PC4′ is selected from proline and leucine; PC5′ is selected from glycine, valine, leucine and isoleucine; and PC6′ is selected from arginine, lysine, citrulline, glutamine, threonine, leucine, norleucine and methionine. The vaccine according to claim 2 , wherein the S of the amphiphile comprises two or more solubilizing groups (SG).

7. The vaccine according to claim 6, wherein the two or more SGs are connected to the rest of the S via a dendrimer amplifier.

8. The vaccine according to any one of claims 6-7, wherein the SG is independently selected from amines, hydroxyls, carboxylic acids and / or sugar molecules, wherein the sugar molecules are independently selected from mannose, glucose, glucosamine, N-acetylglucose, galactose, galactosamine and N-acetylgalactosamine, N-acetylglucosamine, phosphoserine and any derivatives thereof, agonists of CD22a, sialyl Lewis x and combinations thereof.

9. The vaccine according to any one of claims 7-8, wherein the dendrimer amplifier comprises 1 to 10 generations of repeating monomer units, each generation having between 2 and 6 branches.

10. The vaccine according to claim 9, wherein the repeating monomer unit is selected from FG1-(CH2) y2 CH(R 1 )2、FG1-(CH2) y2 C(R 1 )3、FG1-(CH2CH2O) y2 CH(R 1 )2、FG1-(CH2CH2O) y2 C(R 1 )3 and FG1-CH(R 1 )2、FG1-C(R 1 )3, in R 1 is independently selected at each occurrence from (CH2) y3 -FG2, (OCH2CH2) y3 -FG2 and CH2(OCH2CH2) y3 -FG2); y2 and y3 are independently at each occurrence an integer of 1 to 6 repeating units; FG1 is the first functional group; and FG2 is the second functional group. The vaccine according to claim 10 , wherein FG1 is —NH 2 ; and FG2 is —CO 2 H, or FG1 is —CO 2 H; and FG2 is —NH 2 .

12. The vaccine of any one of claims 7 to 11, wherein the dendrimer amplifier comprises a polyethylene oxide (PEG) group.

13. The vaccine according to any one of claims 2 to 12, wherein the H of the amphiphile comprises a higher alkane, an aromatic group, a fatty acid, a sterol, a polyunsaturated hydrocarbon, squalene, a saponin or a polymer.

14. The vaccine according to any one of claims 1 to 13, wherein the H of the peptide antigen conjugate comprises a higher alkane, an aromatic group, a fatty acid, a sterol, a polyunsaturated hydrocarbon or a polymer.

15. The vaccine of claim 13 or 14, wherein each H independently comprises a poly(amino acid) comprising a monomer selected from the group consisting of a hydrophobic amino acid (M), a reactive amino acid (N), a spacer amino acid (O), a charged amino acid (P), and combinations thereof, with the proviso that at least one of M or N is present.

16. The vaccine of claim 15, wherein each H independently comprises a poly(amino acid) having the formula: ， wherein M, N, O and P are each independently present or absent, provided that at least one of M or N is present; m, n, o and p each independently represent an integer from 1 to 100, and the sum of m, n, o and p is less than or equal to 100; R 3 Selected from hydrogen, NH2, NH-CH3, NH-(CH2) y5 CH3, OH or a drug molecule (D) directly or via a suitable linker X; and y5 is an integer selected from 1 to 6.

17. The vaccine of claim 16, wherein P, when present, is , where each R 5 Independently, a group comprising 1 to 2 charged functional groups.

18. The vaccine of claim 17, wherein O, when present, is , wherein each Q is independently selected from (CH2) y6 and (CH2CH2O) y7 CH2CH2; each y6 is independently selected from an integer from 1 to 6; and each y7 is independently selected from an integer from 1 to 4.

19. The vaccine according to any one of claims 16 to 18, wherein N, when present, is , wherein each X1 is independently a suitable linker; and each D is independently a drug molecule.

20. The vaccine according to any one of claims 16 to 19, wherein M, when present, is , where each R 4 are independently hydrophobic groups.

21. The vaccine according to claim 20, wherein R 4 yes in, α is aryl or heteroaryl; X2 is present or absent and, when present, is a suitable linker; y8 is an integer selected from 0 and 6; and Z 1 、Z 2 and Z 3 Each is independently selected from hydrogen, fluorine, hydroxy, amino, alkyl and fluoroalkyl.

22. The vaccine according to claim 20, wherein each R 4 Independently selected from: 、 、 、 、 、 、 、 、 、 、 、 、 and , wherein each X2 is directly linked or independently selected from a suitable linker, and each y8 is independently selected from an integer between 0 and 6.

23. The vaccine according to any one of claims 2 to 22, wherein the vaccine comprises at least one D selected from ATP-competitive mTOR inhibitors; preferably wherein the at least one D is selected from AZD-8055, AZD-2016, KU-0063794, CC223, Torin-1, Torin-2, INK-128, WYE354, WYE132, OSI-027, OXA-01, PI-103, NVP-BEZ235, GNE-493, GSK2126458, rapamycin, tacrolimus, everolimus, RAD001, CCI-779 and AP23573.

24. The vaccine of any one of claims 2-23, wherein B, when present, is a hydrophilic polymer or peptide.

25. The vaccine of any one of claims 2-24, wherein said U of said amphiphile, when present, comprises an amide, a thioether, or a triazole.

26. The vaccine of any one of claims 1-24, wherein the U of the peptide antigen conjugate, when present, comprises an amide, a thioether, or a triazole.

27. The vaccine according to any one of claims 1 to 26, wherein the vaccine comprises a molar ratio of peptide antigen conjugate to amphiphile of between about 4:1 to about 1:20, preferably 1:

1.

28. The vaccine of any one of claims 1-27, wherein the vaccine is a tolerance-inducing allergy vaccine, a tolerance-inducing autoimmune disease vaccine, or a tolerance-inducing transplant rejection vaccine.

29. A method of treating or preventing an autoimmune disease, an allergy, or an infectious disease in a subject in need thereof, the method comprising administering to the subject the vaccine of any one of claims 1-27.

30. The vaccine according to any one of claims 1-27, wherein the at least one peptide antigen conjugate comprises (A) selected from the group consisting of an autoantigen, an alloantigen, and an allergen, said (A) comprising a sequence of 7 to 45 amino acids in length.

31. The vaccine according to any one of claims 1 to 27, wherein at least one peptide antigen (A) comprises alpha-aminobutyric acid and / or norleucine.

32. The vaccine according to any one of claims 1-27, wherein A is selected from the group consisting of: QLQPFPQPELPYPQPQLPYPQPQPFR (SEQ ID NO: 486), PQLPYPQPELPYPQPQPFRPEQPYPQPQP (SEQ ID NO: 487), QGIIQPEQPAQLEVI (SEQ ID NO: 464), PQPQQPEQPFPQPEQEFPQPQQPQQSFPEQQPPL (SEQ ID NO: 488), PQQPFPQPEQPFCQQPQ (SEQ ID NO: 489), QQFLQPEQPFPQQPEQPYPQQPEQPFPQPQQ (SEQ ID NO: 490), QQFSQPEQEFPQPQQPQQSFPEQQPPF (SEQ ID NO: 491), PTPLQPEQPFPQQPQQPQQPFPQPEQPFPWQPQ (SEQ ID NO: 492). NO:492), SSPLQPEQPFPQQPQQPFPEQPQQPQ (SEQ ID NO:493), QSIPQPEQPFPQPEQPFPQSQE (SEQ ID NO:494), PQQPFPQQPQQIIPQ (SEQ ID NO:495), PQQPIPEQPQPYPEQPQPYPQQ (SEQ ID NO: 496), QQPPFSEQEQPVLPQ (SEQ ID NO:484), QPPFSQQQESPFSQQ (SEQ ID NO:485), and PQQPFPQPEQPFBQQPQ (SEQ ID NO:497).

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