Self-assembled nanoparticles

Abstract

The present disclosure relates to vaccines 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, where E1 is an N-terminal extension, E2 is a C-terminal extension, A is a peptide antigen, H is a hydrophobic block, one or more drug molecules (D) are optionally attached to each H directly or via a suitable linker X1, U is a linker, [ ] indicates that the group is optional, and - indicates that two adjacent groups are covalently linked directly to each other or indirectly to each other via a suitable linker X. The vaccines are useful for treating or preventing cancer, autoimmune diseases, allergies, or infectious diseases.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Application No. 63 / 380,931, filed October 25, 2022, the disclosure of which is incorporated herein by reference in its entirety.

[0002] This invention was created in the performance of a Cooperative Research and Development Agreement with the National Institutes of Health, an agency of the Department of Health and Human Services. The United States Government has certain rights in this invention.

[0003] 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 PEGylated peptide-antigen conjugate compositions, processes for formulating drug molecules with PEGylated peptide-antigen conjugate compositions to form nanoparticles, and therapeutic uses of the nanoparticles for drug delivery.

[0004] Reference to sequence listing A Sequence Listing, submitted in XML format, is hereby submitted and incorporated by reference pursuant to U.S.C. § 1.52(e). The ASCII text file name for the Sequence Listing is VNA002WO.xml, the file creation date is October 24, 2023, and the file size is 199 KB. [Background technology]

[0005] Various peptide-based vaccine technologies are known and have been developed for delivering peptide antigens to induce an immune response. Patent Application Publication No. 2020 / 0054741 relates to a novel peptide-based vaccine, a method for producing the novel peptide-based vaccine, and its use for delivering peptide antigens to induce an immune response. Patent Application Publication No. 2021 / 0113705 discloses an improved method for producing a peptide-based vaccine.

[0006] PEGylation has been widely utilized as a means to protect molecules from the immune system. Numerous PEGylated recombinant proteins (Ramos-de-la-Pena, AM, et al. International Journal of Peptide Research and Therapeutics, 2020, 26:333-348) have been approved by the FDA based on improved pharmacokinetics, which has been attributed to improved ability to evade the immune system. PEGylated liposome carriers, including DOXIL, have also been developed based on the similar principle that PEGylation reduces immune recognition. Our previous results using PEGylated peptide antigen conjugates, as disclosed in U.S. Patent Application No. 2020 / 0054741, indicated that such PEGylation is likely to be detrimental to the immune response generated by peptide antigens. Recently, there has been growing interest in the use of peptide-based vaccines to induce tolerance for the treatment of autoimmune conditions. WO 2022 / 177993 relates to a vaccine comprising a novel amphiphilic composition and at least one peptide antigen conjugate having a charged block, wherein the amphiphilic material and / or at least one peptide antigen conjugate comprises a dendron amplifier. A potential problem has been found to be that peptide antigen conjugates having a charged block cause dose-dependent hemolysis of red blood cells. Amphiphilic carriers have been introduced to overcome this problem, but the associated reduction in hydrodynamic stability remains. Therefore, there is currently a need for improved peptide antigen conjugate compositions that maintain hydrodynamic stability while reducing hemolytic activity, as well as improved methods for delivering multiple immunomodulators and peptide antigens in particles to induce tolerance. It is an object of the present disclosure to provide improved compositions, methods for producing vaccines, and uses thereof to induce immune responses that address the above-mentioned challenges. [Prior art documents] [Patent documents]

[0007] [Patent Document 1] U.S. Patent Application Publication No. 2020 / 0054741 [Patent Document 2] International Publication No. 2022 / 177993 [Non-patent literature]

[0008] [Non-Patent Document 1] Ramos-de-la-Pena,AM,et al.International Journal of Peptide Research and Therapeutics,2020,26:333-348 Summary of the Invention [Means for solving the problem]

[0009] The present disclosure provides novel PEGylated peptide-antigen conjugate compositions that can be used to form nanoparticles, including micellar structures or polymersomes, methods for making PEGylated peptide-antigen conjugate compositions, processes for formulating drug molecules with PEGylated peptide-antigen conjugate compositions to form nanoparticles, and therapeutic uses of the nanoparticles for drug delivery. In a first aspect of the present disclosure, there is provided 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, During the ceremony, A is a peptide antigen, E1 is the N-terminal extension, E2 is a C-terminal extension, H, independently at each occurrence, is a hydrophobic block; and one or more drug molecules (D) are optionally attached to each H, either directly or via a suitable linker, X1; U is, independently at each occurrence, a linker; [ ] means that the group is optional; - indicates that two adjacent groups are directly linked to each other by a covalent bond, or indirectly linked to each other via a suitable linker X.

[0010] In a second aspect of the present disclosure, there is provided 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, During the ceremony, A is a peptide antigen, E1 is the N-terminal extension, E2 is a C-terminal extension, H, independently at each occurrence, is a hydrophobic block; and one or more drug molecules (D) are optionally attached to each H, either directly or via a suitable linker, X1; S is a solubilizing block, B is a spacer, U is, independently at each occurrence, a linker; [ ] means that the group is optional; - indicates that two adjacent groups are directly linked to each other by a covalent bond, or indirectly linked to each other via a suitable linker X.

[0011] In one embodiment of the vaccine, the amphiphile S comprises a dendron amplifier.

[0012] In a third aspect of the present disclosure, there is provided a vaccine for inducing tolerance 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, During the ceremony, A is a peptide antigen, E1 is the N-terminal extension, E2 is a C-terminal extension, H, independently at each occurrence, is a hydrophobic block; and one or more drug molecules (D) are optionally attached to each H, either directly or via a suitable linker, X1; S is a solubilizing block, B is a spacer, U is, independently at each occurrence, a linker; [ ] means that the group is optional; - denotes that two adjacent groups are directly linked to each other by a covalent bond or indirectly linked to each other via a suitable linker X; wherein the amphiphilic material comprises a dendron amplification material; The at least one peptide antigen A is selected from an autoantigen, an alloantigen and an allergen.

[0013] In a fourth aspect of the present disclosure, there is provided herein 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; During the ceremony, A is a peptide antigen, E1 is the N-terminal extension, E2 is a C-terminal extension, H, independently at each occurrence, is a hydrophobic block; and one or more drug molecules (D) are optionally attached to each H, either directly or via a suitable linker, X1; S is a solubilizing block, B is a spacer, U is, independently at each occurrence, a linker; [ ] means that the group is optional; - denotes that two adjacent groups are directly linked to each other by a covalent bond or indirectly linked to each other via a suitable linker X; wherein the amphiphilic material comprises a dendron amplification material; wherein at least one A comprises a sequence in which one or more cysteine ​​residues are replaced with alpha-aminobutyric acid and / or one or more methionine residues are replaced with norleucine.

[0014] In a fifth aspect of the present disclosure, there is provided a vaccine comprising at least one peptide antigen (A), wherein at least one A comprises a sequence in which one or more cysteine ​​residues are replaced with alpha aminobutyric acid and / or one or more methionine residues are replaced with norleucine.

[0015] In one embodiment of the present disclosure, a vaccine is provided, wherein at least one peptide antigen (A) comprises alpha aminobutyric acid and / or norleucine.

[0016] In a sixth aspect of the present disclosure, there is provided a vaccine for inducing tolerance 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, During the ceremony, A is a peptide antigen, E1 is the N-terminal extension, E2 is a C-terminal extension, H, independently at each occurrence, is a hydrophobic block; and one or more drug molecules (D) are optionally attached to each H, either directly or via a suitable linker, X1; S is a solubilizing block, B is a spacer, U is, independently at each occurrence, a linker; [ ] means that the group is optional; - denotes that two adjacent groups are directly linked to each other by a covalent bond or indirectly linked to each other via a suitable linker X; wherein the amphiphilic material comprises a dendron amplification material; At least one A is selected from an autoantigen, an alloantigen and an allergen, and at least one D is present.

[0017] In one embodiment of a vaccine for inducing tolerance, at least one D is selected from the group consisting of inhibitors of mTOR, RORgt, CDK8 / 19, and HDAC, as well as inhibitors of AHR, RAR, and A 2a The agonist is selected from the group consisting of:

[0018] In another embodiment of the vaccine for inducing tolerance, at least one D is selected from ATP-competitive mTOR inhibitors.

[0019] In some embodiments of a vaccine for inducing tolerance, 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.

[0020] 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: A is a peptide antigen, E1 is the N-terminal extension, E2 is a C-terminal extension, H, independently at each occurrence, is a hydrophobic block; and one or more drug molecules (D) are optionally attached to each H, either directly or via a suitable linker, X1; U is, independently at each occurrence, a linker; [ ] means that the group is optional; - indicates that two adjacent groups are directly bonded to each other by a covalent bond, or indirectly bonded to each other via a suitable linker.

[0021] In an eighth aspect of the present disclosure, there is provided herein 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 PEG-[E1]-A-[E2]-[U]-H and H-[U]-[E1]-A-[E2]-PEG and an amphiphile having the formula S-[B]-[U]-H; During the ceremony, A is a peptide antigen, E1 is the N-terminal extension, E2 is a C-terminal extension, H, independently at each occurrence, is a hydrophobic block; and one or more drug molecules (D) are optionally attached to each H, either directly or via a suitable linker, X1; S is a solubilizing block, B is a spacer, U is, independently at each occurrence, a linker; [ ] means that the group is optional; - indicates that two adjacent groups are directly bonded to each other by a covalent bond or indirectly bonded to each other via a suitable linker; wherein the amphiphilic material comprises a dendron amplifier material and the at least one peptide antigen is selected from a self-antigen and a foreign antigen.

[0022] In one embodiment of the method of treating an autoimmune disease, the vaccine is administered intravenously, subcutaneously, or intramuscularly.

[0023] In a ninth aspect of the present disclosure, there is provided herein a method for improving vaccine efficacy and / or tolerability, the method comprising administering to a subject 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; During the ceremony, E1 is the N-terminal extension, E2 is a C-terminal extension, H, independently at each occurrence, is a hydrophobic block; and one or more drug molecules (D) are optionally attached to each H, either directly or via a suitable linker, X1; S is a solubilizing block, B is a spacer, U is, independently at each occurrence, a linker; [ ] means that the group is optional; - indicates that two adjacent groups are directly bonded to each other by a covalent bond or indirectly bonded to each other via a suitable linker; wherein the amphiphilic material comprises a dendron amplifier material and the at least one peptide antigen is selected from a self-antigen and a foreign antigen.

[0024] In a tenth aspect of the present disclosure, provided herein are methods for preparing peptide-antigen conjugates having the formula PEG-[E1]-A-[E2]-[U]-H and H-[U]-[E1]-A-[E2]-PEG.

[0025] In an eleventh aspect of the present disclosure, there is provided herein a method of preparing a vaccine comprising a peptide antigen conjugate of formula PEG-[E1]-A-[E2]-[U]-H and an amphiphile having formula S-[B]-[U]-H, During the ceremony, A is a peptide antigen, E1 is the N-terminal extension, E2 is a C-terminal extension, H, independently at each occurrence, is a hydrophobic block; and one or more drug molecules (D) are optionally attached to each H, either directly or via a suitable linker, X1; S is a solubilizing block, B is a spacer, U is, independently at each occurrence, a linker; [ ] means that the group is optional; - indicates that two adjacent groups are directly linked to each other by a covalent bond, or indirectly linked to each other via a suitable linker X.

[0026] In a twelfth aspect of the present disclosure, there is provided herein a vaccine for inducing tolerance comprising at least one peptide antigen conjugate having a formula selected from 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, and E2 is a C-terminal extension; H, independently at each occurrence, is a hydrophobic block; and at least one drug molecule (D) is optionally attached to each H, either directly or via a suitable linker, X1; U is, independently at each occurrence, a linker; [ ] means that the group is optional; - indicates that two adjacent groups are directly linked to each other by a covalent bond, or indirectly linked to each other via a suitable linker X.

[0027] In some embodiments of a vaccine for inducing tolerance, when at least one peptide antigen has a grand mean hydropathy value of >0 and / or a mean peptide antigen solubility in aqueous solution at pH 5.5-8.5 of <1 mg / mL, the vaccine further comprises an amphiphile having the formula S-[B]-[U]-H, where S is a solubilizing block; B is a spacer, H is a hydrophobic block, U is a linker, [ ] means that the group is optional; - denotes that two adjacent groups are directly linked to each other by a covalent bond or indirectly linked to each other via a suitable linker X; Here, the amphiphilic material S comprises a dendron amplifier.

[0028] In some embodiments of a vaccine for inducing tolerance, the vaccine does not contain an amphiphile having the formula S-[B]-[U]-H.

[0029] In some embodiments of a vaccine for inducing tolerance, if at least one peptide antigen has a grand mean hydropathy value of ≦0 and / or a mean peptide antigen solubility in aqueous solution at pH 5.5-8.5 of ≧1 mg / mL, the vaccine does not contain an amphiphile having the formula S-[B]-[U]-H.

[0030] In some embodiments of a vaccine for inducing tolerance, the vaccine comprises at least one D selected from an ATP-competitive mTOR inhibitor, 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.

[0031] In some embodiments of the vaccine for inducing tolerance, at least one D is covalently linked to a hydrophobic block (H) directly or indirectly via a linker X1.

[0032] In some embodiments of the vaccine for inducing tolerance, the linker X1 comprises an amide, carbamate, hydrazone, ketal, or silyl ether moiety.

[0033] In some specific embodiments of the vaccine for inducing tolerance, the linker X1 comprises a degradable peptide comprising 2 to 6 amino acids.

[0034] In some embodiments of the vaccine for inducing tolerance, the linker X1 comprising an enzyme-degradable peptide comprises an amino acid residue P1 selected from arginine, lysine, acetyl lysine, boc-protected lysine, citrulline, glutamine, threonine, leucine, norleucine, alpha-aminobutyric acid, and methionine, and an amino acid residue P2 selected from beta-alanine, glycine, serine, leucine, valine, and isoleucine.

[0035] In a thirteenth aspect of the present disclosure, there is provided herein 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: E1 is the N-terminal extension, E2 is a C-terminal extension, H, independently at each occurrence, is a hydrophobic block; and one or more drug molecules (D) are optionally attached to each H, either directly or via a suitable linker, X1; U is, independently at each occurrence, a linker; [ ] means that the group is optional; - denotes that two adjacent groups are directly linked to each other by a covalent bond or indirectly linked to each other via a suitable linker X; A is 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), PTPLQPE The peptide antigen is selected from the group consisting of QPFPQQPQQPQQPFPQPEQPFPWQPQ (SEQ ID 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).

[0036] In one embodiment of a vaccine of the present disclosure, a 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) that is water soluble to at least 1 mg / mL, or wherein the peptide antigen conjugate exhibits a tendency to aggregate after 24 hours at room temperature when the total peptide antigen conjugate concentration is ≧0.5 mM in an aqueous formulation buffer containing ≦20% organic solvent.

[0037] In some embodiments of the vaccines of the present disclosure, the micelles are about 5 nm to about 50 nm in diameter, or about 10 nm to about 30 nm in diameter.

[0038] In one embodiment of the vaccine of the present disclosure, at least one drug molecule (D) is non-covalently associated with the micelle.

[0039] In a fourteenth aspect of the present disclosure, there is provided 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: A is a peptide antigen, E1 is the N-terminal extension, E2 is a C-terminal extension, H, independently at each occurrence, is a hydrophobic block; and one or more drug molecules (D) are optionally attached to each H, either directly or via a suitable linker, X1; U is, independently at each occurrence, a linker; [ ] means that the group is optional; - denotes that two adjacent groups are directly linked to each other by a covalent bond or indirectly linked to each other via a suitable linker X; Provided herein are vaccine formulations comprising compositions with a formulation buffer comprising 10% DMSO (v / v) in phosphate buffered saline at pH 7.4 or tris(hydroxymethyl)aminomethane in saline (0.9% NaCl) at pH 6.5-8.5.

[0040] In one embodiment of the vaccine formulation, a non-ionic surfactant is further included.

[0041] In some embodiments of the vaccine formulation, the non-ionic surfactant is selected from polysorbate-20 and sodium dodecyl sulfate. [Brief explanation of the drawings]

[0042] [Figure 1] Figures A-B show the turbidity (Figure 1A) and particle size (Figure 1B) of vaccine formulations containing peptide-antigen conjugates with different N-terminal groups, i.e., charged block (C) or PEG. See Table 1C and the Experimental Section for a description of materials and methods. [Figure 2]We demonstrate that replacing the positively charged block (C) of a peptide antigen conjugate of formula C-E1-A-E2-UH with a PEG group to produce a conjugate of formula PEG-E1-A-E2-UH abrogates the dose-dependent hemolytic activity associated with the charged block (see Table 2B and Experimental Section for a description of Materials and Methods). [Figure 3] Figures A to C show the effect of the N-terminal group of the peptide antigen conjugate on tolerability after intravenous administration. These data indicate that vaccine formulations containing peptide antigen conjugates of the formula PEG-E1-A-E2-UH were better tolerated after intravenous administration than peptide antigen conjugates of the formula C-E1-A-E2-UH. Figure A shows the experimental design and vaccination schedule. Figure B shows body weight dynamics using body weight normalized to day 0. Figure C shows survival rates with Kaplan-Meier curves. See Table 3B and the Experimental section for a description of materials and methods. [Figure 4A] Figure 4 shows the kinetics of disability scores for mice with experimental autoimmune encephalomyelitis (EAE) given different treatments (Table 4) following EAE disease induction at time 0. The experimental design in terms of EAE induction and treatment days is shown. [Figure 4B] Figure 1 shows the kinetics of the lesion score for mice with experimental autoimmune encephalomyelitis (EAE) given different treatments (Table 4) following EAE disease induction at time 0. Figure 2 shows the kinetics of the lesion score for the intravenous route group. [Figure 4C] Figure 4 shows the kinetics of lesion scores for mice with experimental autoimmune encephalomyelitis (EAE) given different treatments (Table 4) following EAE disease induction at time 0. Figure 5 shows the kinetics of lesion scores for the subcutaneous route group. These data indicate that treatment with a vaccine composition comprising a peptide antigen conjugate of formula PEG-E1-A-E2-UH or C-E1-A-E2-UH ameliorates the disease, but that the peptide antigen conjugate of formula PEG-E1-A-E2-UH provided superior efficacy compared to C-E1-A-E2-UH when administered by the SC route. [Figure 5]Figures 5A-B show the effects of different treatments (Table 4) on T cell phenotype in mice with experimental autoimmune encephalomyelitis (EAE). These 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 percentage of CD4 T cells expressing IFN-gamma (Th1 CD4 T cells, Figure 5A) and IL-17 (Th17 CD4 T cells, Figure 5B) compared with untreated animals (Group 1), whereas peptide-antigen conjugates of the formula PEG-E1-A-E2-UH resulted in a greater reduction in IFN-gamma-producing cells compared with C-E1-A-E2-UH. An asterisk (*) indicates p<0.05 for a Student's t-test comparing stimulated and unstimulated samples. [Figure 6A] Figure 5 shows the kinetics of disability scores for mice with experimental autoimmune encephalomyelitis (EAE) given different treatments (Table 5) following EAE disease induction on days 0 and 28. An experimental design including multiple EAE induction and treatment days is shown. [Figure 6B] Figure 1 shows the kinetics of the disability score for mice with experimental autoimmune encephalomyelitis (EAE) given different treatments (Table 5) following EAE disease induction on days 0 and 28. Figure 1 shows the kinetics of the disability score for the group given treatment by the intramuscular route (IM). [Figure 6C] Figure 1 shows the kinetics of the lesion score for mice with experimental autoimmune encephalomyelitis (EAE) given different treatments (Table 5) following EAE disease induction on days 0 and 28. The kinetics of the lesion score for the group given treatment by intravenous route (IV) is shown. [Figure 6D]Table 5 shows the kinetics of lesion scores for mice with experimental autoimmune encephalomyelitis (EAE) given different treatments (Table 5) following EAE disease induction on days 0 and 28. Table 5 shows the kinetics of lesion scores for IM-treated mice after a second induction of EAE. These data indicate that treatment with a vaccine composition comprising a peptide antigen conjugate of formula PEG-E1-A-E2-UH or C-E1-A-E2-UH ameliorates the disease, but that the peptide antigen conjugate of formula PEG-E1-A-E2-UH provides superior efficacy compared to C-E1-A-E2-UH, with PEG-E1-A-E2-UH showing comparable efficacy via both the IM and IV routes. [Figure 7] Figures 7A-B show the effects of different treatments (Table 5) on T cell phenotype in mice with experimental autoimmune encephalomyelitis. These 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 percentage of CD4 T cells expressing IFN-gamma (Th1 CD4 T cells, Figure 7A) and IL-17 (Th17 CD4 T cells, Figure 7B) compared with untreated animals (Group 1), and vaccines containing Torin reduced the percentage of CD4 T cells expressing IFN-gamma compared with animals treated with vaccines containing rapamycin (Group 7). An asterisk (*) indicates p<0.05 for a Student's t-test comparing stimulated and unstimulated samples. [Figure 8] Panels A to B show the turbidity at 1 hour (hours) and 24 hours (hours) (hours) for representative vaccines (Table 8) with varying storage temperatures and peptide-antigen conjugate concentrations. Panel B shows the filtration recovery rate for representative vaccines (Table 8) with varying storage temperatures and peptide-antigen conjugate concentrations. 24 hours (hours) after formulation and storage, each vaccine composition was filtered through a 0.2 μm filter, and the area under the curve at 220 nm was then assessed by HPLC, where the pre-filtration AUC was divided by the post-filtration AUC. [Figure 9]A shows the turbidity at 1 hour (hours (hr)) and 24 hours (hours (hr)) for representative vaccines (Table 10) that varied in peptide antigen conjugate (PAC) formula, surfactant identity and concentration, amphiphile presence, and formulation method. B shows the filtration recovery at 24 hours (hours (hr)) for representative vaccines (Table 10) that varied in PAC formula, surfactant identity and concentration, amphiphile presence, and formulation method. [Figure 10] A shows the turbidity at 1 hour, 24 hours, and 48 hours at room temperature (approximately 23°C) for representative vaccines (Table 13) that varied in PAC formula, surfactant identity, presence of amphiphile, and formulation method. B shows the filtration recovery at 24 hours and 48 hours at room temperature (approximately 23°C) for representative vaccines (Table 13) that varied in PAC formula, emulsifier identity, presence of amphiphile, and formulation method. [Figure 11] A shows turbidity at 24 and 48 hours at room temperature (approximately 23°C) for representative vaccines varying mTORi identity, mTORi molar ratio, PAC concentration, and formulation buffer (Table 14). B shows filtration recovery at 1 hour, 24 hours, and 48 hours at room temperature (approximately 23°C) for representative vaccines varying mTORi identity, mTORi molar ratio, PAC concentration, and formulation buffer (Table 14). [Figure 12] A shows the turbidity at 24 and 48 hours at room temperature (approximately 23°C) for representative vaccines with varying mTORi identity, mTORi molar ratio, and PAC concentration (Table 15). B shows the filtration recovery at 1 hour, 24 hours, and 48 hours at room temperature (approximately 23°C) for representative vaccines with varying mTORi identity, mTORi molar ratio, and PAC concentration (Table 15). [Figure 13A] Figure 17 shows the kinetics of disability scores for mice with experimental autoimmune encephalomyelitis (EAE) given different treatments (Table 17) following EAE disease induction on day 0, followed by treatment on days 0, 7, and 14. The experimental design including EAE induction and treatment days is shown. [Figure 13B]Figure 17 shows the kinetics of lesion scores for mice with experimental autoimmune encephalomyelitis (EAE) given different treatments (Table 17) on days 0, 7, and 14 following EAE disease induction. Figure 17 shows the kinetics of lesion scores for the group given treatment by intramuscular route (IM). It shows that incorporation of an irrelevant peptide antigen (CPNE1) does not affect efficacy. [Figure 13C] Figure 17 shows the kinetics of the lesion score for mice with experimental autoimmune encephalomyelitis (EAE) given different treatments (Table 17) following EAE disease induction on day 0, followed by treatment on days 0, 7, and 14. Figure 17 shows the kinetics of the lesion score for the group given treatment by intramuscular route (IM), demonstrating that incorporation of Torin-1 or rapamycin improves efficacy. [Figure 13D] Table 17 shows the kinetics of lesion scores for mice with experimental autoimmune encephalomyelitis (EAE) given different treatments (Table 17) on days 0, 7, and 14 following EAE disease induction on day 0. Table 17 shows the kinetics of lesion scores for groups given treatment by the intramuscular route (IM). Table 17 shows the kinetics of lesion scores for groups given treatment by the intramuscular route (IM). Table 17 shows that incorporation of rapamycin at a molar ratio of 1:1 drug molecule to peptide antigen conjugate demonstrated the highest efficacy compared to no rapamycin incorporation or a molar ratio of 0.5:1 rapamycin to peptide antigen conjugate. These data demonstrate that treatment with a vaccine composition comprising a peptide antigen conjugate of the formula PEG-E1-A-E2-UH provides protection from the development of EAE disease, but that a peptide antigen conjugate of the formula PEG-E1-A-E2-U-H+D results in superior efficacy compared to the same formulation lacking the mTOR inhibitor. [Figure 14A] Figure 18 shows the kinetics of disability scores for mice with experimental autoimmune encephalomyelitis (EAE) given different treatments (Table 18) following EAE disease induction on day 0 followed by treatment on days 0, 7, and 14. The experimental design including EAE induction and treatment days is shown. [Figure 14B]Table 18 shows the kinetics of lesion scores for mice with experimental autoimmune encephalomyelitis (EAE) treated with different treatments (Table 18) on days 0, 7, and 14 following EAE disease induction on day 0. Table 18 shows the kinetics of lesion scores for the group treated intramuscularly (IM) at a dose of 2.5 nmol of peptide antigen. At equivalent doses, the four formulations tested showed virtually equal efficacy. The data show that a formulation composed of PEG-E1-A-E2-U-H is equivalent to or superior in efficacy to formulations composed of either (i) E1-A-E2-U-H + SB-[U]-H, (ii) PEG-E1-A-E2-U-H + SB-[U]-H + surfactant, or (iii) PEG-E1-A-E2-U-H + SB-[U]-H. [Figure 14C] Table 18 shows the kinetics of lesion scores for mice with experimental autoimmune encephalomyelitis (EAE) treated with different treatments (Table 18) on days 0, 7, and 14 following EAE disease induction on day 0. Table 18 shows the kinetics of lesion scores for groups treated intramuscularly (IM) at a dose of 10 nmol of peptide antigen. At equivalent doses, the four formulations tested showed virtually equal efficacy. The data show that a formulation composed of PEG-E1-A-E2-UH is equivalent to or superior in efficacy to formulations composed of either (i) E1-A-E2-U-H + SB-[U]-H, (ii) PEG-E1-A-E2-U-H + SB-[U]-H + surfactant, or (iii) PEG-E1-A-E2-U-H + SB-[U]-H. [Figure 14D]Table 18 shows the kinetics of lesion scores for mice with experimental autoimmune encephalomyelitis (EAE) treated with different treatments (Table 18) on days 0, 7, and 14 following EAE disease induction on day 0. Table 18 shows the kinetics of lesion scores for groups treated intramuscularly (IM) with a dose of 40 nmol of peptide antigen. At equivalent doses, the four formulations tested demonstrated virtually equal efficacy. The data show that a formulation composed of PEG-E1-A-E2-UH is equivalent to or superior in efficacy to formulations composed of either (i) E1-A-E2-U-H + SB-[U]-H, (ii) PEG-E1-A-E2-U-H + SB-[U]-H + surfactant, or (iii) PEG-E1-A-E2-U-H + SB-[U]-H. [Figure 14E] Figure 18 shows the kinetics of the disability score for mice with experimental autoimmune encephalomyelitis (EAE) given different treatments (Table 18) following EAE disease induction on day 0, followed by treatment on days 0, 7, and 14. A comparison of the efficacy of PEG-El-A-E2-UH formulations at doses of 2.5, 10, or 40 nmol, respectively, on EAE disease score is shown. [Figure 14F] Table 18 shows the kinetics of the disability score for mice with experimental autoimmune encephalomyelitis (EAE) given different treatments (Table 18) on days 0, 7, and 14 following EAE disease induction on day 0. Statistical evaluation between groups (one-way ANOVA with Tukey's correction for multiple comparisons; asterisks represent p-values ​​≦0.0001) was used to assess the area under the curve for EAE disease scores for days 7 through 28. These data demonstrate a strong relationship between the dose of peptide antigen construct administered and the EAE disease score. [Figure 15A] Table 19 shows the kinetics of disability scores and the area under the curve (AUC) of disease scores for mice with experimental autoimmune encephalomyelitis (EAE) for mice receiving different treatments (Table 19) on days 0, 7, and 14 following EAE disease induction on day 0. The experimental design, including EAE induction and treatment days, is shown. Groups

[12] and

[13] received treatment only on day 0. [Figure 15B]Figure 1 shows the kinetics of disability scores for mice with experimental autoimmune encephalomyelitis (EAE) for mice given different treatments (Table 19) following EAE disease induction on day 0, followed by treatment on days 0, 7, and 14. A comparison of the effectiveness of EAE disease scores for IM or SC administered mice where treatment was administered either three times or only once on day 0 is shown. [Figure 15C] Figure 1 shows the area under the curve (AUC) of disease scores for mice with experimental autoimmune encephalomyelitis (EAE) for mice given different treatments (Table 19) following EAE disease induction on day 0, followed by treatment on days 0, 7, and 14. Evaluation of the area under the curve of EAE disease scores for days 7-21 is shown. [Figure 16A]

[0033] Figure 2 shows the experimental design for testing GLU peptide-antigen conjugates in Sprague Dawley rats using the formulations (Table 21). The experimental design 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]

[0033] Figure 2 shows the immunological results of a study of GLU peptide antigen conjugates in Sprague Dawley rats using the formulations (Table 21). Anti-gliadin antibody titers are shown on day 35 of the study in both gliadin-sensitized and non-sensitized (naive) rats. [Figure 16C]

[0033] Figure 2 shows immunological results of a study of GLU peptide-antigen conjugates in Sprague Dawley rats using formulations (Table 21). Activation levels of phosphorylated S6 are assessed in antigen-presenting cells (RT1B+, CD3-) isolated from blood 4 hours after treatment on study day 0. [Figure 16D]

[0033] Figure 2 shows immunological results of a study of GLU peptide-antigen conjugates in Sprague Dawley rats using the formulations (Table 21). The frequency (%) of IFN-γ cells among the CD4+ cell population isolated from blood on day 35 is shown. [Figure 16E]2 shows immunological results of a study of GLU peptide-antigen conjugates in Sprague Dawley rats using the formulations (Table 21). The frequency (%) of IL-17+ cells among the CD4+ cell population isolated from blood on day 35 is shown. [Figure 16F]

[0033] Figure 2 shows immunological results of a study of GLU peptide-antigen conjugates in Sprague Dawley rats using the formulations (Table 21). The frequency (%) of IFN-γ+ cells among the CD8+ cell population isolated from blood on day 35 is shown. [Figure 16G]

[0033] Figure 2 shows immunological results of a study of GLU peptide-antigen conjugates in Sprague Dawley rats using the formulations (Table 21). The frequency (%) of IL-17+ cells among the CD8+ cell population isolated from blood on day 35 is shown. [Figure 17A] Figure 2 shows the results of an in vitro mTOR inhibitor assay in splenocyte populations following in vitro stimulation with LPS. Splenocytes from C57BL / 6 mice were dissociated into single-cell suspensions and plated in vitro for stimulation. Test article compounds were added to the cells at 0.5, 5, 50, or 500 nM concentrations for 1 hour, after which lipopolysaccharide (LPS) was added to the cells for 2 hours to stimulate mTOR activity and downstream phosphorylation of S6 to phosphorylated S6 (pS6). Cells were then fixed and stained for flow cytometry analysis. Test article compounds (Table 24) were resuspended in PBS before being added to splenocyte cultures. mTOR inhibition, as measured by the pS6 assay, using small molecule drug analogs of everolimus with conjugable handles is shown. [Figure 17B]Figure 2 shows the results of an in vitro mTOR inhibitor assay in splenocyte populations following in vitro stimulation with LPS. Splenocytes from C57BL / 6 mice were dissociated into single-cell suspensions and plated in vitro for stimulation. Test article compounds were added to the cells at 0.5, 5, 50, or 500 nM concentrations for 1 hour, after which lipopolysaccharide (LPS) was added to the cells for 2 hours to stimulate mTOR activity and downstream phosphorylation of S6 to phosphorylated S6 (pS6). Cells were then fixed and stained for flow cytometry analysis. Test article compounds (Table 24) were resuspended in PBS before being added to splenocyte cultures. mTOR inhibition, measured by the pS6 assay, is shown using a peptide antigen conjugate of an everolimus derivative conjugated to an H-block. These two examples demonstrate that everolimus or rapamycin can be modified to be conjugated to other molecules, and that the conjugated versions of the mTOR inhibitors remain active when conjugated via either an amide bond (group [7]) or a cleavable bond (group [8]). [Figure 17C] Figure 2 shows the results of an in vitro mTOR inhibitor assay in splenocyte populations following in vitro stimulation with LPS. Splenocytes from C57BL / 6 mice were dissociated into single-cell suspensions and plated in vitro for stimulation. Test article compounds were added to the cells at 0.5, 5, 50, or 500 nM concentrations for 1 hour, after which lipopolysaccharide (LPS) was added to the cells for 2 hours to stimulate mTOR activity and downstream phosphorylation of S6 to phosphorylated S6 (pS6). Cells were then fixed and stained for flow cytometry analysis. Test article compounds (Table 24) were resuspended in PBS before being added to splenocyte cultures. mTOR inhibition, as measured by the pS6 assay, using small molecule analogs of Torin-1 is shown. DETAILED DESCRIPTION OF THE INVENTION

[0043] The above-mentioned aspects, as well as other aspects, features, and advantages of the present disclosure, are described below in connection with various embodiments and with reference to the accompanying figures. definition For the purpose of guiding those skilled in the art in the practice of the present disclosure, details of terms and methods are given below to provide greater clarity regarding the compounds, compositions, methods and their use(s). It is understood that the terms in this disclosure are useful for the purpose of providing a better description of certain embodiments and should not be considered limiting.

[0044] About: In the context of this disclosure, "about" when referring to a measurable value, such as an amount, time duration, etc., is intended to encompass variations of ±20%, ±10%, ±5%, ±1%, or ±0.1% from the specified value, where such variations are appropriate for practicing 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.

[0045] Administration: Providing or giving a pharmaceutical agent, e.g., an immunogenic composition comprising an amphiphilic block copolymer described herein and a drug(s), to a subject by any effective route. Exemplary routes of administration include, but are not limited to, oral, injection (such as subcutaneous, intramuscular, intradermal, intraperitoneal, and intravenous), transdermal, topical, intranasal, vaginal, and inhalation routes.

[0046] "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 described herein. The compound or composition can be administered to a subject by another person, or it can be self-administered by the subject.

[0047] Antigen: Any molecule containing an epitope capable of binding to a T cell or B cell receptor and stimulating an immune response, particularly a B cell response and / or a T cell response, in a subject. An epitope may include a peptide, glycopeptide, lipid, or any suitable molecule containing an epitope capable of interacting with a specific B cell or T cell receptor component. Such an interaction may result in a response by the immune cell. "Epitope" refers to the region of a peptide antigen with which a B cell and / or T cell protein, i.e., a B cell receptor and a T cell receptor, interacts. Antigens used in embodiments of the present disclosure may be selected from pathogens, cancerous cells, autoantigens, alloantigens, or allergens. Many such antigens may be used in accordance with the inventive embodiments of the present disclosure and are discussed in more detail throughout the specification.

[0048] Antigen-presenting cell (APC): Any cell that presents antigen bound to an MHC class I or class II molecule to a T cell, including but not limited to monocytes, macrophages, dendritic cells, B cells, T cells, and Langerhans cells.

[0049] Amphiphilic: The term "amphiphilic" is used herein to mean a substance that contains both hydrophilic or polar groups and hydrophobic groups.

[0050] 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 expresses CD4, and these cells are commonly referred to as helper T cells or CD4 T cells.

[0051] 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 expresses CD8, and these cells are commonly referred to as cytotoxic T cells (CTLs), killer T cells, or CD8 T cells.

[0052] 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 from molecules that do not carry a full integer charge, such as polar molecules. Two charged molecules with similar charges repel each other, while two charged molecules with different charges attract each other. Charge is often described as a positive or negative integer. A molecular charge can be easily estimated based on the molecule's Lewis structure and accepted methods known to those skilled in the art. Charge can arise from inductive effects; for example, atoms bonded together with different electron affinities can result in a polar covalent bond, resulting in a partially negatively charged atom and a partially positively charged atom. For example, the bonding of nitrogen 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 charge if the number of electrons assigned to that atom is less than or equal to the atom's atomic number. 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 may be permanently charged or may have a charge depending on pH. A charged functional group may have a partial or complete integer charge, which may be positive or negative, and 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 may 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 may include a positive functional group, a negative functional group, or both a positive and a negative functional group. The net charge of a charged group may be positive, negative, or neutral. A charged monomer refers to a monomer that contains a charged group. A charged amino acid is a type of charged monomer. NOTE: The net charge of particles containing amphiphile and / or peptide-antigen conjugates that further contain charged groups, e.g., charged monomers such as charged amino acids, can be estimated by summing the charges of each functional group within the amphiphile and / or peptide-antigen conjugate. The charged block (C) is a type of solubilization block.

[0053] Click chemistry reaction: a bioorthogonal reaction that joins two compounds together under mild conditions in a high-yield reaction that produces minimal biocompatible and / or harmless by-products. 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.

[0054] Copolymer: A polymer derived from two (or more) different monomers, as opposed to homopolymers, in which only one monomer is used. Because copolymers contain at least two types of constitutional units (also known as structural units), copolymers can be classified based on how these units are arranged along the chain. A copolymer may be a statistical (or random) copolymer, in which the two or more monomer units are randomly distributed; a copolymer may be an alternating copolymer, in which two or more monomer units are distributed in an alternating sequence; or, for example, a copolymer, such as a poly(amino acid), may be produced by solid-phase peptide synthesis (SPPS) and have a specific order of monomer units. The term "block copolymer" refers generically to a polymer composed of two or more consecutive blocks of different constitutional monomers or comonomers (where the blocks contain two or more different monomers). A block copolymer may be used herein to refer to a copolymer comprising two or more homopolymer subunits, two or more copolymer subunits, or one or more homopolymer subunits and one or more copolymer subunits, where the subunits may be directly linked by a covalent bond or the subunits may be indirectly linked through an intermediate non-repeating subunit, such as a junction block or linker. The blocks may be based on linear and / or brush structures. Block copolymers having two or three distinct blocks are referred to herein as "diblock copolymers" and "triblock copolymers," respectively. Copolymers may be collectively referred to as polymers; for example, statistical copolymers may be referred to as polymers or copolymers. Similarly, block copolymers may be collectively referred to as polymers. As used herein, a copolymer refers to a polymer containing two or more types of monomers, while a terpolymer is a copolymer having three monomer units.

[0055] Critical micelle concentration (CMC): refers to the concentration of a material above which micelles spontaneously form to satisfy thermodynamic equilibrium.

[0056] Drug: refers to any pharmaceutically active molecule, including but not limited to proteins, peptides, sugars, saccharides, nucleosides, inorganic compounds, lipids, nucleic acids, small synthetic chemical compounds, macrocyclic compounds, etc., that has a physiological effect when ingested or otherwise introduced into the body. Pharmaceutically active compounds include, for example, analgesics, anesthetics, anti-inflammatory agents, anthelmintics, antiarrhythmic agents, antiasthmatic agents, antibiotics (including penicillin), anticancer agents, anticoagulants, antidepressants, antidiabetics, antiepileptics, antihistamines, antitussives, antihypertensives, antimuscarinics, antimycobacterial agents, antitumor agents, antioxidants, antipyretics, immunosuppressants, immunostimulants, antithyroid agents, antivirals, anxiolytics and sedatives (hypnotics and neuroleptics), astringents, bacteriostatics, beta-adrenergic receptor blocking agents, blood products and blood substitutes, bronchodilators, buffers, cardiac inotropes, chemotherapeutic agents, contrast media, corticosteroids, antitussives (expectorants and mucolytics), diagnostic agents, imaging diagnostics. Drugs may be selected from a variety of known classes of compounds, including anti-inflammatory drugs, diuretics, dopamine agonists (anti-Parkinson's agents), free radical scavengers, growth factors, hemostatic agents, immunological agents, lipid regulating agents, muscle relaxants, proteins such as therapeutic antibodies and antibody fragments, MHC-peptide complexes, cytokines and growth factors, glycoproteins, peptides and polypeptides, parasympathomimetics, parathyroid calcitonin, biphosphonates, prostaglandins, radiopharmaceuticals, hormones, sex hormones (including steroids), anti-allergic agents, stimulants and appetite suppressants, steroids, sympathomimetics, thyroid agents, vaccines, vasodilators, and xanthines. Drugs may also be referred to as pharmaceutically active agents, pharmaceutically active substances, or biologically active compounds or bioactive molecules. Any drug molecule in the formulas described herein is abbreviated as "D."

[0057] Drug delivery: The method or process of administering a pharmaceutical compound to achieve a therapeutic effect in a human or animal.

[0058] Effective amount: An amount of a compound, material, or composition effective to achieve a particular biological result, such as, but not limited to, a biological result disclosed, described, or exemplified herein. Such a result may include, but is not limited to, an effective reduction in symptoms associated with any of the conditions referred to herein, as determined by any means suitable in the art.

[0059] Hydropathy index / GRAVY value: A numerical value representing the hydrophobic or hydrophilic properties of an amino acid or amino acid sequence. There are various scales that can be used to describe the relative hydrophobic and hydrophilic properties of the amino acids that make up a peptide. In the present disclosure, the Kyte and Doolittle hydropathy scale (Kyte J, Doolittle RF, J. Mol. Biol. 157:105-32, 1983) is used to calculate the grand average hydropathy value (GRAVY) (sometimes referred to as the GRAVY score). The GRAVY value of a peptide is the sum of the hydropathy values ​​of all 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 higher the GRAVY value, the more hydrophobic the peptide sequence is considered to be, while the lower the GRAVY value, the more hydrophilic the peptide sequence is considered to be.

[0060] Hydrophilicity: Refers to the tendency of a material to disperse or solubilize freely 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 interactions 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 not depending on what it is being compared to. Hydrophilic molecules are often polar and / or charged and have good aqueous solubility, e.g., soluble at concentrations of at least 1.0 mg / mL or higher. A hydrophilic group refers to a portion of a molecule that is polar and / or charged and has good aqueous solubility.

[0061] Hydrophobicity: Refers to a material's tendency to avoid contact with water. A material is considered hydrophobic if it prefers to interact with other hydrophobic materials and avoids interactions with hydrophilic materials. Hydrophobicity is a relative term, and the same molecule may be described as hydrophobic or not depending on what it is being compared to. Hydrophobic molecules are often non-polar and uncharged and poorly soluble in water, e.g., insoluble in water, or soluble in water only at concentrations below 1 mg / mL, typically 0.1 mg / mL or less, or more preferably 0.01 mg / mL or less. Hydrophobic monomers are monomers, e.g., hydrophobic amino acids, that contain hydrophobic groups and form polymers that are insoluble in water or insoluble in water at certain temperatures, pHs, and salt concentrations. The hydrophobic group refers to the portion of a molecule that is hydrophobic. 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 or that are soluble only at a concentration of about 1.0 mg / mL or less in aqueous solution at a pH of about pH 7.4. Amphipathic drugs are drug molecules that have a tendency to organize 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.

[0062] Immune response: A change in the activity of a cell of the immune system, such as a B cell, T cell, or monocyte, as a result of stimulation, either directly or indirectly, such as through cell or cytokine mediation. In certain embodiments, the response is specific for a particular antigen (an "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. Such an immune response may result in the production of additional T cell progeny and / or T cell migration. In other embodiments, the response is a B cell response, resulting 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. An antigen can be used to stimulate an immune response, leading to the activation of cytotoxic T cells that kill virally infected or cancerous cells. In other embodiments, an antigen can be used to induce tolerance or immunosuppression. A tolerogenic response can result from the unresponsiveness of T cells or B cells to an antigen. Suppressive immune responses can result from the priming and / or activation of regulatory cells, such as regulatory T cells, or the transdifferentiation of effector cells into regulatory cells that downregulate, i.e., attenuate, the immune response.

[0063] Immunogenic composition: A formulation of material that includes an antigen and optionally an immunomodulatory agent that induces a measurable immune response to the antigen. For example, a vaccine is a type of immunogenic composition.

[0064] Immunomodulator: Refers to a class of drugs that modulate the activity of cells of the immune system, including immunostimulants and immunosuppressants.

[0065] Immunostimulator: refers to any synthetic or naturally occurring drug that promotes proinflammatory and / or cytotoxic activity by immune cells. Exemplary immunostimulators include pattern recognition receptor (PRR) agonists, such as synthetic or naturally occurring agonists of toll-like receptors (TLRs), stimulator of interferon genes agonists (STINGa), nucleotide-binding oligomerization domain-like receptor (NLR) agonists, retinoic acid-inducible gene-I-like receptor (RLR) agonists, and certain C-type lectin receptors (CLRs), as well as 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 by binding to stimulatory receptors, such as anti-CD40; or certain antibodies, antibody fragments, or synthetic peptides that activate immune cells by blocking inhibitory receptors, such as anti-CTLA4 and anti-PD1. Various immunostimulants suitable for practicing the present disclosure are described throughout this specification. For clarity, certain pharmaceutically active compounds that stimulate the immune system may be referred to as immunostimulants, or more generally as drug molecules (abbreviated as "D" herein).

[0066] Immunosuppressant: refers to any synthetic or naturally occurring drug that suppresses proinflammatory and / or cytotoxic activity by immune cells or the humoral immune system, e.g., antibodies and complement proteins. Immunosuppressants may mediate their effects through one or more of the following mechanisms of action: priming suppressor cells, e.g., regulatory T cells; killing, inhibiting, or inactivating proinflammatory, cytotoxic, and / or B cells; transdifferentiating proinflammatory and / or cytotoxic T cells into suppressor cells; and / or by sequestering and / or limiting the mobility of proinflammatory, cytotoxic, and / or B 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; PD1 agonists; and other molecules that suppress the pro-inflammatory or cytotoxic activity of immune cells or antibodies.Various immunosuppressants suitable for implementing the present disclosure are described throughout this specification, including Treg-promoting immunomodulators.For clarity, immunosuppressants can be more generally referred to as drug molecules (abbreviated as "D" in the formula).

[0067] In vivo delivery: Administration of a composition, such as a composition comprising an amphiphilic block copolymer and a drug(s), to a subject by topical, transdermal, suppository (rectal, vaginal), pessary (vaginal), intravenous, oral, subcutaneous, intraperitoneal, intrathecal, intramuscular, intracranial, inhalation, oral, or any other suitable route.

[0068] Linked or Bonded: The terms "linked" and "bonded" mean joined together, either directly or indirectly. A first moiety may be covalently or non-covalently linked to a second moiety. In some embodiments, a first molecule is covalently linked to another molecule. In some embodiments, a first molecule is linked to another molecule by electrostatic attraction. In some embodiments, a first molecule is linked to another molecule by dipole-dipole forces (e.g., hydrogen bonds). In some embodiments, a first molecule is linked to another molecule by van der Waals forces (also known as London forces). A first molecule may be linked to another molecule by any and all combinations of such bonds. Molecules may be indirectly linked, such as by using a linker (sometimes referred to as a linker molecule). Molecules may be indirectly linked through the intermediation of a component that independently binds both molecules non-covalently. As used in the formulas herein, the term "linker," sometimes abbreviated as "X," refers to any suitable linker molecule. Certain preferred linkers may be designated by other symbols such as X1, X2, X3, X4, X5 and U. A variety of linkers are described throughout the specification.

[0069] A "bilayer membrane" or "bilayer(s)" is a self-assembled film of amphiphiles or supraamphiphiles in aqueous solution.

[0070] Micelle: A spherical receptacle with a single monolayer that defines a closed compartment. Generally, amphiphilic molecules spontaneously form micellar structures in polar solvents. In contrast to bilayers, such as liposomal bilayers, micelles are "faceted" in that they present a protruding hydrophilic polar outer surface and a hydrophobic inner surface.

[0071] 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, of which the density (or "mol %) of monomer A equals 10 mol %, has 10 monomer units of A, and the remaining 90 monomer units (or "monomers") may be monomer B or another monomer, unless otherwise specified.

[0072] Monomeric unit: The term "monomeric unit" or "monomer unit" is used herein to mean a unit of a polymer molecule that contains the same or a similar number of atoms as one of the monomers. A monomeric unit, as used herein, can be of a single type (homogeneous) or of various types (heterogeneous). For example, a poly(amino acid) contains monomeric units of amino acids. A monomeric unit may also be referred to as a monomer or a monomer unit, etc.

[0073] Net Charge: The total electrostatic charge carried by a molecule, or, where specified, by a portion or segment of a molecule.

[0074] Particle: A nano- or micro-sized supramolecular structure composed of an assembly of molecules. For example, the amphiphile and peptide antigen conjugates of the present disclosure form particles in aqueous solution. In some embodiments, particle formation by the amphiphile and / or peptide antigen conjugate is pH or temperature dependent. In some embodiments, nanoparticles composed of the amphiphile and / or peptide antigen conjugate have an average diameter of 5 nanometers (nm) to 500 nm. In some embodiments, nanoparticles composed of the amphiphile and / or peptide antigen conjugate form micelles and have an average diameter of 5 nanometers (nm) to 50 nm, e.g., 10 to 30 nm. In some embodiments, nanoparticles composed of the amphiphile and / or peptide antigen conjugate may be larger than 100 nm.

[0075] Pattern recognition receptors (PRRs): Receptors expressed by various cell populations, particularly innate immune cells, that bind to a diverse group of synthetic and naturally occurring molecules. Several classes of PRRs exist. 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 called immunostimulants 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.

[0076] Peptide or Polypeptide: Two or more natural or unnatural amino acid residues linked together sequentially by one or more amide bonds. The amino acid residues may contain post-translational modification(s) (e.g., glycosylation, citrullination, homocitrullination, oxidation, and / or phosphorylation). Such modifications may mimic post-translational modifications that occur naturally in vivo or may be unnatural. Any one or more of the components of the amphiphile and / or peptide antigen conjugate may comprise a peptide.

[0077] Peptide modifications: Peptides can be altered or otherwise synthesized with one or more of the several modifications described below. Furthermore, analogs (non-peptide organic molecules), derivatives (chemically functionalized peptide molecules obtained starting from peptides), and variants (homologs) of these peptides can be utilized in the methods described herein. The peptides described herein include amino acid sequences, analogs, derivatives, and variants, which may be either L- and / or D-versions. Unless otherwise specified, any peptide sequence referred to herein contains L-amino acids, preferably only L-amino acids. Such peptides can include naturally occurring and non-naturally occurring peptides, analogs, derivatives, and variants.

[0078] Peptides can be modified by any of a variety of chemical techniques to produce derivatives that have similar activity to the unmodified peptide, and optionally, other desirable properties. For example, the carboxylic acid group of the peptide, whether at the carboxyl terminus or in a side chain, can be provided in the form of a salt of a pharmaceutically acceptable cation, or esterified to form a compound such as CC1-CC2. 16Esters (where CC refers to a carbon chain (hence CC1 refers to a single carbon and CC16 refers to 16 carbons)) can be formed or converted to amides. The amino group of the peptide, whether at the amino terminus or in the side chain, can be in the form of a pharmaceutically acceptable acid addition salt, such as HCl, HBr, trifluoroacetate, formate, benzoate, toluenesulfonate, maleate, tartrate, and other organic salts, or can be modified or converted to an amide, for example, by acetylation.

[0079] The peptides may be modified to contain substituents that contain a positive charge or a negative charge, or both. The positive and / or negative charges may be affected by the pH at which the peptide is present.

[0080] Hydroxyl groups on peptide side chains are cleaved from C1 to C6 using well-recognized techniques. 16 Alkoxy or C1-C 16 The phenyl and phenol rings of the peptide side chains may be converted to esters, or the hydroxyl groups may be converted to introduce a negative charge (e.g., sulfated or phosphorylated). The phenyl and phenol rings of the peptide side chains may be substituted with one or more halogen atoms, such as fluorine, chlorine, bromine, or iodine, or with C1-C 16 Alkyl, C1-C 16The functional groups may be substituted with alkoxy, carboxylic acids and their esters, or amides of such carboxylic acids. Methylene groups in peptide side chains can be extended to analogous C2-C4 alkylenes. Thiols can be used to form disulfide bonds or thioethers, for example, by reaction with maleimide. Thiols can be protected with any of several well-recognized protecting groups, such as acetamide groups. Those skilled in the art will also recognize methods for introducing cyclic structures into the peptides of the present invention to provide selected conformational constraints and thereby improve stability. For details of additional modifications that can be made to functional groups, see Greene et al., "Greene's Protective Groups in Organic Synthesis," Fourth Edition, John Wiley & Sons, Inc. 2006.

[0081] Cysteine ​​residues of naturally occurring peptide antigens can be replaced with alpha-aminobutyric acid or serine, and methionine residues with norleucine to obtain non-naturally occurring peptide antigens that induce immune responses that are cross-reactive with the naturally occurring peptide antigens. Preferred methods for preparing and using peptide antigens with non-native sequences are described throughout this specification and in WO2022 / 177993, which is incorporated herein by reference.

[0082] Pharmaceutically acceptable vehicles: Pharmaceutically acceptable vehicles (or carriers) useful in the present disclosure include conventional carriers, excipients, and diluents. Remington's Pharmaceutical Sciences, by E. W. Martin, Mack Publishing Co., Easton, PA, 15th Edition (1975), describes compositions and formulations suitable for pharmaceutical delivery of one or more therapeutic compositions, such as one or more therapeutic cancer vaccines and additional pharmaceutical agents.

[0083] 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 preferred embodiments, when such pharmaceutical compositions are intended for human administration, particularly via invasive routes of administration (i.e., routes such as injection or implantation that bypass transport or diffusion across epithelial barriers), the aqueous solutions are pyrogen-free or substantially pyrogen-free. Excipients can be selected, for example, to provide delayed release of the drug or to selectively target one or more cells, tissues, or organs. Pharmaceutical compositions can be in unit dosage forms such as tablets, capsules (including sprinkle capsules and gelatin capsules), granules, lyophiles for reconstitution, powders, liquids, syrups, suppositories, injectables, and the like. The compositions can also be present in transdermal delivery systems, e.g., skin patches. The compositions can also be present in liquid formulations suitable for topical administration, such as ointments or creams.

[0084] Pharmaceutically acceptable carriers may contain physiologically acceptable agents that act, for example, to stabilize, increase the solubility, or increase the absorption of a compound such as a compound of the present invention. 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 selection of a pharmaceutically acceptable carrier, including a physiologically acceptable agent, depends, for example, on the route of administration of the composition. The pharmaceutical composition preparation can be a self-emulsifying or self-microemulsifying drug delivery system. The pharmaceutical composition (preparation) can also be, for example, a liposome or other polymer matrix that can have a compound of the present invention incorporated therein. For example, liposomes containing phospholipids or other lipids are non-toxic, physiologically acceptable, and metabolizable carriers that are relatively simple to prepare and administer.

[0085] Polarity: A description of a property of a substance. Polarity is a relative term and can describe molecules or portions of molecules that have a partial charge resulting from differences in electronegativity between atoms bonded together within a molecule, such as the bond between nitrogen and hydrogen. Polar molecules prefer to interact with other polar molecules and typically do not associate with non-polar molecules. In specific, non-limiting examples, polar groups can contain hydroxyl groups, or amino groups, or carboxyl groups, or charged groups. In specific, non-limiting examples, polar groups can prefer to interact with polar solvents such as water. In specific, non-limiting examples, the introduction of additional polar groups can increase the solubility of portions of a molecule.

[0086] Polymer: A molecule containing repeating structural units (monomers). As described in more detail throughout this disclosure, polymers may be used in any number of components of amphiphiles, peptide-antigen conjugates, and drug molecule conjugates, and may 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 broadly to encompass molecules containing as few as three or more monomers, which may sometimes be referred to as oligomers.

[0087] Polymerization: A chemical reaction, usually carried out with a catalyst, heat, or light, in which monomers combine to form linear, branched, or crosslinked macromolecules (polymers). Chain, branched, or crosslinked macromolecules can be further modified by additional chemical synthesis using appropriate substituents and chemical reactions. Polymerization generally occurs 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, generating a free electron on the other side. This free electron then reacts with another monomer, causing the chain to become self-propagating, thus 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 cleavage from a water molecule. In other forms of polymerization, monomers are added to a growing chain one at a time by the stepwise introduction of activated monomers, such as during solid-phase peptide synthesis (SPPS).

[0088] Polymersomes: vesicles assembled from synthetic multiblock polymers in aqueous solution. Unlike liposomes, polymersomes do not contain lipids or phospholipids as their predominant components. As a result, polymersomes can be thermally, mechanically, and chemically unique, and in particular, potentially more durable and resilient than even the most stable lipid vesicles. Polymersomes organize during the swelling process of lamellar structures, for example, by film or bulk rehydration, as described below, or through an additional migration step, or by other known methods. Like liposomes, polymersomes form by "self-assembly," a spontaneous, entropy-driven process that prepares a closed, semipermeable membrane.

[0089] Purified: A substance or composition that is relatively free of impurities or materials that degrade or contaminate the substance or composition. The term purified is a relative term and does not require absolute purity. Substantial purification refers to purification from impurities. A substantially purified substance or composition is typically at least 60%, 70%, 80%, 90%, 95%, 98%, or 99% pure.

[0090] Soluble: Able to be molecularly or ionically dispersed in a solvent to form a homogeneous solution. When referring to amphiphiles, peptide-antigen conjugates, drug molecule conjugates, and / or drug molecules, soluble is understood to be a single molecule in solution that does not organize into multimers or other supramolecular structures due to hydrophobic or other non-covalent interactions. Soluble molecules are understood to be freely dispersed as single molecules in solution. The hydrophobic block (H) described herein is insoluble or only soluble up to a concentration of about 0.1 mg / mL or less. Solubility can be determined by visual inspection, turbidity measurement, or dynamic light scattering.

[0091] Subject and patient: These terms may be used interchangeably herein to refer to both humans and non-human animals, including avian and non-human mammals, such as rodents (e.g., mice and rats), non-human primates (e.g., rhesus monkeys), companion animals (e.g., domestic dogs and cats), livestock (e.g., pigs, sheep, cattle, llamas, and camels), and non-domesticated animals (e.g., big cats).

[0092] Targeting molecule: Broadly defined as a molecule that directs a drug molecule to a specific tissue or cell population. Targeting molecules are defined by their intended use and therefore include structurally diverse molecules, including, but not limited to, antibodies, Fabs, peptides, aptamers, saccharides (e.g., saccharides that bind to lectin receptors and / or are recognized by cellular transporters), amino acids, neurotransmitters, etc. Targeting molecules are often selected from molecules that bind to cellular receptors that can activate downstream signaling cascades and / or affect the activity of other linked molecules, and therefore are often classified as drug molecules (D) in the present disclosure. Additionally, targeting molecules can also have a solubilizing effect and can be considered either or both of a drug molecule (D) group and / or a solubilizing (SG) group.

[0093] T cell: A type of white blood cell that is part of the immune system and can participate in immune responses. T cells include, but are not limited to, CD4 T cells and CD8 T cells. CD4 T cells present the CD4 glycoprotein on their surface, and these cells are often referred to as helper T cells. These cells often orchestrate immune responses, including antibody responses and cytotoxic T cell responses, although CD4 T cells (e.g., regulatory T cells) can also suppress immune responses, or CD4 T cells can act as cytotoxic T cells. CD8 T cells present the CD8 glycoprotein on their surface, and these cells are often referred to as cytotoxic or killer T cells, although CD8 T cells can also suppress immune responses.

[0094] Treating, preventing, or ameliorating a disease: "Treating" refers to an intervention that reduces signs or symptoms or markers of a disease or pathological condition after it has begun to develop. For example, treating a disease may result in a reduction in tumor burden, i.e., a decrease in the number or size of tumors and / or metastases, or treating a disease may result in immune tolerance, which reduces systems associated with an autoimmune condition. "Preventing" a disease refers to preventing the disease from developing completely. A disease may be prevented from developing completely. A disease may be prevented from progressing, such as in severity or extent. "Ameliorating" refers to a reduction in the number or severity of signs or symptoms or markers of a disease, such as cancer.

[0095] Reducing the signs or symptoms or markers of disease or disease-related pathological conditions refers to any observable beneficial effect of treatment and / or any observable effect on proximal surrogate endpoints, such as tumor volume, regardless of whether it is symptomatic. Reducing the signs or symptoms associated with tumors or viral infections can be evidenced, for example, by delaying the onset of clinical symptoms of disease in susceptible subjects (such as subjects with tumors that have not yet metastasized, or subjects who may be exposed to viral infections), or by reducing the severity of some or all clinical symptoms of disease, slowing the progression of disease (for example, by extending the lifespan of subjects with tumors or viral infections), reducing the number of disease recurrences, improving the overall health or well-being of subjects, or other parameters well known in the art (for example, specific to specific tumors or viral infections). "Preventive" treatment is a treatment administered to subjects who do not show signs of disease or who only show early signs, with the aim of reducing the risk or severity of developing pathology.

[0096] Tumor or cancer or neoplasm: An abnormal growth of cells, which may be benign or malignant, that often, but not always, causes clinical symptoms. "Neoplastic" cell growth refers to cell growth that is unresponsive to physiological cues such as growth factors and inhibitory factors.

[0097] A "tumor" is a collection of neoplastic cells. In most cases, tumor refers to a collection of neoplastic cells that form 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.

[0098] Cancer refers to the malignant growth of neoplastic cells, which can be either solid or liquid. Characteristics of cancer that define it as malignant include metastasis, interference with the normal function of nearby cells, release of abnormal levels of cytokines or other secretory products, and suppression or exacerbation of inflammatory or immune response(s), infiltration of nearby or distant tissues or organs such as lymph nodes, etc.

[0099] Tumors that do not produce substantial adverse clinical symptoms and / or grow slowly are termed "benign."

[0100] "Malignant" means causing or likely to cause significant clinical symptoms. A tumor is said to be "malignant" if it causes substantial clinical symptoms by invading surrounding tissues and / or metastasizing and / or by producing and secreting chemical mediators that have effects on nearby or distant body systems.

[0101] "Metastatic disease" refers to cancer cells that have left the original tumor site and traveled to other parts of the body, for example, through the bloodstream, through the lymphatic system, or through a body cavity such as the abdominal or thoracic cavity.

[0102] 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 often assessed by physical examination, radiography, or pathological examination.

[0103] An "established" or "pre-existing" tumor is one that is present at the time treatment is initiated. In many cases, an established tumor can be identified by a diagnostic test. In some embodiments, an established tumor can be palpated. In some embodiments, an established tumor is at least 500 mm 3 , e.g., at least 600 mm 3 , at least 700mm 3 , or at least 800mm 3 In other embodiments, the tumor is at least 1 cm in size. With respect to solid tumors, established tumors generally have a newly established, robust blood supply and may induce regulatory T cells (Tregs) and myeloid-derived suppressor cells (MDSCs).

[0104] Unit dose: A discrete amount of a pharmaceutical composition containing a predetermined amount of an active ingredient.

[0105] Vesicle: A fluid-filled sac. In some embodiments, the vesicle is a sac containing an amphiphile. In some embodiments, the sac is a nanoparticle-based vesicle, which refers to a vesicle with a size or dimension in the nanometer range. In some embodiments, the polymeric vesicle is a vesicle formed from one or more polymers.

[0106] Definition: As used herein, the term "optionally" or "optionally" means that the subsequently described event or circumstance may or may not occur, and that the description includes both cases where the event or circumstance occurs and cases where it does not occur. For example, "optionally substituted alkyl" refers to an alkyl that may or may not be substituted. It is understood that the substituents and substitution patterns on the compounds of the present invention can be selected by one skilled in the art to result in chemically stable compounds that can be readily synthesized from readily available starting materials by techniques known in the art, as well as the methods described below. When a substituent is itself substituted with more than one group, it is understood that these multiple groups can be on the same carbon or on different carbons, so long as a stable structure results. As used herein, the term "optionally substituted" refers to the replacement of 1 to 6 hydrogen radicals in a given structure with the radical of the specified substituent, including, but not limited to, hydroxyl, hydroxyalkyl, alkoxy, halogen, alkyl, nitro, silyl, acyl, acyloxy, aryl, cycloalkyl, heterocyclyl, amino, aminoalkyl, cyano, haloalkyl, haloalkoxy, -OCO-CH-O-alkyl, -OP(O)(O-alkyl) or -CH-OP(O)(O-alkyl). Preferably, "optionally substituted" refers to the replacement of 1 to 4 hydrogen radicals in a given structure with a substituent as described above. More preferably, 1 to 3 hydrogen radicals are replaced by a substituent as described above. It is understood that a substituent may be further substituted. As used herein, the term "alkyl" refers to a C1-C 10 Straight chain alkyl group or C1-C 10 "Alkyl" refers to saturated aliphatic groups, including, but not limited to, branched alkyl groups. Preferably, "alkyl" refers to a C1-C6 straight chain alkyl group or a C1-C6 branched chain alkyl group. Most preferably, "alkyl" refers to a C1-C4 straight chain alkyl group or a C1-C4 branched chain alkyl group. 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, neo-pentyl, 1-hexyl, 2-hexyl, 3-hexyl, 1-heptyl, 2-heptyl, 3-heptyl, 4-heptyl, 1-octyl, 2-octyl, 3-octyl, or 4-octyl. "Alkyl" groups may be optionally substituted. The term "acyl" is art-recognized and refers to a group represented by the general formula hydrocarbylC(O)-, preferably alkylC(O)-. 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 hydrocarbylC(O)NH-. The term "acyloxy" is art-recognized and refers to a group represented by the general formula hydrocarbylC(O)O-, preferably alkylC(O)O-. 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. The term "alkoxyalkyl" refers to an alkyl group substituted with an alkoxy group and may be represented by the general formula alkyl-O-alkyl. The term "alkyl" refers to saturated aliphatic groups, including straight-chain alkyl groups, branched-chain alkyl groups, cycloalkyl (alicyclic) groups, alkyl-substituted cycloalkyl groups, and cycloalkyl-substituted alkyl groups. In preferred embodiments, a straight-chain or branched-chain alkyl has 30 or fewer carbon atoms in its backbone (e.g., C for straight chain).1-30 , C for branched chain 3-30 ), more preferably having 20 or fewer carbon atoms. Furthermore, the term "alkyl," as used throughout the specification, examples, and claims, is intended to include both unsubstituted and substituted alkyl groups, the latter of which refers to alkyl moieties in which substituents replace a hydrogen on one or more carbons of the hydrocarbon backbone, including haloalkyl groups such as trifluoromethyl and 2,2,2-trifluoroethyl. "C" when used with chemical moieties such as acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy. x-y " or "C x ~C y The term "alkyl" is intended to include groups containing x to y carbons in the chain. C0 alkyl indicates a hydrogen if the group is in a terminal position and a bond if it is internal. For example, C 1-6 Alkyl groups contain 1 to 6 carbon atoms in the chain. The term "alkylamino," as used herein, refers to an amino group substituted with at least one alkyl group. The term "alkylthio," as used herein, refers to a thiol group substituted with an alkyl group and may be represented by the general formula alkylS-. As used herein, the term "amide" refers to the group [ka] In the formula, R 22 and R 23 each independently represents hydrogen or a hydrocarbyl group, or R 22 and R 23 together with the N atom to which they are attached complete a heterocycle having 4 to 8 atoms in the ring structure. The terms "amine" and "amino" are art-recognized and refer to both unsubstituted and substituted amines and their salts, e.g., [ka] wherein R 22 , R 23 , and R24 each independently represents hydrogen or a hydrocarbyl group, or R 22 and R 23 together with the N atom to which they are attached complete a heterocycle having 4 to 8 atoms in the ring structure. As used herein, the term "aminoalkyl" refers to an alkyl group substituted with an amino group. As used herein, the term "aralkyl" refers to an alkyl group substituted with an aryl group. The term "aryl" as used herein includes substituted or unsubstituted aromatic carbocycles as well as heteroaryls. The term "aryl" is used interchangeably with the term "aromatic group" herein. Unless otherwise expressly defined herein, aryl moieties independently include alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl, trifluoromethoxy, nitro, trimethylsilanyl, -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 (wherein t is 1 or 2), -S(O) t OR a (wherein t is 1 or 2), -S(O)t N(R a )2 (wherein t is 1 or 2), or PO3(R a )2, wherein each R a are independently hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkyl, heteroaryl, or heteroarylalkyl. Aromatic carbocycles include monocyclic aromatic groups in which each atom of the ring is carbon. Preferably, the ring is a 5- to 7-membered ring, more preferably a 6-membered ring. The term "aryl" also includes polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjacent rings, at least one of which is aromatic; for example, the other cyclic rings may be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclyl. Aryl groups include benzene, naphthalene, phenanthrene, phenol, aniline, and the like. The term "carbamate" is art-recognized and [ka] In the formula, R 22 and R 23 independently represent hydrogen or a hydrocarbyl group. The term "carbocyclylalkyl," as used herein, refers to an alkyl group substituted with a carbocyclic group. The term "carbocycle" includes 5- to 7-membered monocyclic rings and 8- to 12-membered bicyclic rings. Each ring in a bicyclic carbocycle can be selected from saturated, unsaturated, and aromatic rings. Carbocycles include 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 in a fused carbocycle can be selected from saturated, unsaturated, and aromatic rings. For example, an aromatic ring, such as phenyl, can be fused to a saturated or unsaturated ring, such as cyclohexane, cyclopentane, or cyclohexene. Any combination of saturated, unsaturated, and aromatic bicyclic rings, valence permitting, is included in the definition of carbocycle. 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 adamantane. Exemplary fused carbocycles include decalin, 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 one or more positions capable of bearing a hydrogen atom. The term "carbocyclylalkyl," as used herein, refers to an alkyl group substituted with a carbocyclic group. The term "carbonate" is art-recognized and refers to the group --OCO.sub.2--. The term "carboxy," as used herein, refers to a group represented by the formula -CO2H. As used herein, the term "ester" refers to an ester of -C(O)OR 22 In the formula, R 22 represents a hydrocarbyl group. The term "ether" as used herein refers to a hydrocarbyl group linked to another hydrocarbyl group via an oxygen atom. Thus, the ether substituent of a hydrocarbyl group can be hydrocarbyl-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. The terms "halo" and "halogen" as used herein mean halogen and include chloro, fluoro, bromo, and iodo. The terms "hetaralkyl" and "heteroaralkyl," as used herein, refer to an alkyl group substituted with a hetaryl group. The terms "heteroaryl" and "hetaryl" include substituted or unsubstituted aromatic monocyclic ring structures, preferably 5- to 7-membered rings, more preferably 5- or 6-membered rings, which ring structures contain at least one heteroatom, preferably 1 to 4 heteroatoms, more preferably 1 or 2 heteroatoms. The terms "heteroaryl" and "hetaryl" also include polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjacent rings, and 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 groups include, for example, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyridine, pyrazine, pyridazine, pyrimidine, and the like. The term "heteroatom" as used herein means an atom of any element other than carbon or hydrogen. Preferred heteroatoms are nitrogen, oxygen, and sulfur. The term "heterocyclylalkyl," as used herein, refers to an alkyl group substituted with a heterocycle group. 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, which ring structures contain at least one heteroatom, preferably 1 to 4 heteroatoms, more preferably 1 or 2 heteroatoms. The terms "heterocyclyl" and "heterocyclic" also include polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjacent rings, and at least one of the rings is heterocyclic, e.g., 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. As used herein, the term "hydrocarbyl" refers to a group that does not have =0 or =5 substituents, typically has at least one carbon-hydrogen bond, and a primarily carbon backbone, but may optionally contain heteroatoms, bonded through carbon atoms. Thus, groups such as methyl, ethoxyethyl, 2-pyridyl, and even trifluoromethyl are considered hydrocarbyl for purposes of this application, while substituents such as acetyl (which has an =0 substituent on the connecting carbon) and ethoxy (which is connected through an oxygen rather than a carbon) are not considered hydrocarbyl. Hydrocarbyl groups include, but are not limited to, aryl, heteroaryl, carbocyclic, heterocyclic, alkyl, alkenyl, alkynyl, and combinations thereof. As used herein, the term "hydroxyalkyl" refers to an alkyl group substituted with a hydroxy group. The term "lower" when used in conjunction with chemical moieties such as acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy, is intended to include groups in which there are 10 or fewer atoms, preferably 6 or fewer atoms, in the substituent. For example, "lower alkyl" refers to an alkyl group containing 10 or fewer carbon atoms, preferably 6 or fewer carbon atoms. In certain embodiments, an acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy substituent defined herein is a lower acyl, lower acyloxy, lower alkyl, lower alkenyl, lower alkynyl, or lower alkoxy, respectively, whether they appear alone or in combination with other substituents, as in the recitation of hydroxyalkyl and aralkyl (where, for example, atoms in an aryl group are not counted when counting carbon atoms in an alkyl substituent). The terms "polycyclyl," "polycycle," and "polycyclic" refer to two or more rings (e.g., cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclyl) in which two or more atoms are common to two adjacent rings, e.g., the rings are "fused rings." Each of the rings of a polycycle can be substituted or unsubstituted. In certain embodiments, each ring of a polycycle contains 3 to 10 atoms, preferably 5 to 7 atoms, within the ring. The term "sulfate" is art-recognized and refers to the group --OSO.sub.3H, or a pharmaceutically acceptable salt thereof. The term "sulfonamide" is art-recognized and [ka] In the formula, R 22 and R 23 independently represent hydrogen or hydrocarbyl. The term "sulfoxide" is art-recognized and refers to the group --S(O)--. The term "sulfonate" is art-recognized and refers to the group SO3H, or a pharmaceutically acceptable salt thereof. The term "sulfone" is art-recognized and refers to the group -S(O)2-. The term "substituted" refers to a moiety in which a substituent replaces a hydrogen on one or more backbone carbons. It is understood that "substituted" or "substituted with" includes the implicit proviso that such substitution complies with the permissible valences of the substituted atom and substituent, and that the substitution results in a stable compound, e.g., a compound that does not undergo spontaneous transformation by rearrangement, cyclization, elimination, and the like. As used herein, the term "substituted" is intended to include all permissible substituents of organic compounds. In a broad aspect, the permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and nonaromatic substituents of organic compounds. The permissible substituents can be one or more and the same or different for appropriate organic compounds. For purposes of this invention, heteroatoms, such as nitrogen, can have hydrogen substituents and / or any permissible substituents of organic compounds described herein that satisfy the valences of the heteroatoms. Substituents may include any of the substituents described herein, for example, halogen, hydroxyl, carbonyl (such as carboxyl, alkoxycarbonyl, formyl, or acyl), thiocarbonyl (such as thioester, thioacetate, or thioformate), alkoxyl, 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. Those skilled in the art will understand that the moieties substituted on the hydrocarbon chain can themselves be substituted, if appropriate. The term "thioalkyl," as used herein, refers to an alkyl group substituted with a thiol group. As used herein, the term "thioester" refers to a thioester of -C(O)SR 22 or -SC(O)R22 In the formula, R 22 represents a hydrocarbyl. The term "thioether" as used herein is equivalent to an ether where the oxygen has been replaced with a sulfur. The term "urea" is art-recognized and [ka] where R 22 and R 23 independently represent hydrogen or hydrocarbyl. The term "aromatic amino acid" includes amino acids with a side chain containing an aromatic group, 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 typical aromatic amino acids.

[0107] Those of skill in the art will recognize that the definitions provided above are not intended to include impermissible substitution patterns (e.g., methyl substituted with five different groups, etc.). Such impermissible substitution patterns are readily recognized by those of skill in the art. Any functional group disclosed herein and / or defined above can be substituted or unsubstituted, unless otherwise indicated herein. Unless otherwise explained, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. The term "comprises" means "includes." Thus, comprising "A" or "B" refers to 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 provided for illustrative purposes. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present disclosure, suitable methods and materials are described herein. In case of conflict, the present specification, including explanations of terms, will control. Further, the materials, methods, and examples are illustrative only and not intended to be limiting.

[0108] Description of the embodiment Provided herein are compositions of particles comprising an amphiphile and a drug molecule useful for treating or preventing a disease, such as cancer(s), autoimmune disease(s), allergy(s), and / or infectious disease(s). Particles comprising certain compositions of amphiphile and peptide antigen conjugates have particular utility for use as vaccines to treat or prevent a disease, for example, to prevent or treat cancer(s), autoimmune disease(s), allergy(s), and / or infectious disease(s).

[0109] The present disclosure relates to 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, During the ceremony, A is a peptide antigen, E1 is the N-terminal extension, E2 is a C-terminal extension, H, independently at each occurrence, is a hydrophobic block; and one or more drug molecules (D) are optionally attached to each H, either directly or via a suitable linker, X1; U is, independently at each occurrence, a linker; [ ] means that the group is optional; - indicates that two adjacent groups are directly linked to each other by a covalent bond, or indirectly linked to each other via a suitable linker X.

[0110] In embodiments of the vaccine, an amphiphile is present if either (i) the average water 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.

[0111] The present disclosure relates to 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, During the ceremony, A is a peptide antigen, E1 is the N-terminal extension, E2 is a C-terminal extension, H, independently at each occurrence, is a hydrophobic block; and one or more drug molecules (D) are optionally attached to each H, either directly or via a suitable linker, X1; S is a solubilizing block, B is a spacer, U is, independently at each occurrence, a linker; [ ] means that the group is optional; - indicates that two adjacent groups are directly linked to each other by a covalent bond, or indirectly linked to each other via a suitable linker X.

[0112] In one embodiment of the vaccine, the PEG group of the peptide antigen conjugate comprises a terminal functional group selected from OH, MeO-, and NH2.

[0113] In certain embodiments of the vaccine, the PEG group of the peptide antigen conjugate is polyethylene glycol.

[0114] In some embodiments of the vaccine, the PEG group of the peptide antigen conjugate comprises between 4 and 36 monomer units.

[0115] In certain embodiments of the vaccine, the PEG group of the peptide antigen conjugate comprises 4 to 12 monomer units, or 12 to 36 monomer units, preferably 24 monomer units.

[0116] In some embodiments of the vaccine, when an amphiphile is present, the amphiphile comprises a dendron amplifier.

[0117] In some embodiments of the vaccine, the amphiphile S comprises a dendron amplifier, hi other embodiments, the amphiphile S has a dendritic structure.

[0118] In some embodiments of the vaccine, the amphiphile S comprises two or more solubilizing groups (SGs). In other embodiments, two or more SGs are connected to the remainder of S by a dendron amplifier, e.g., 4 to 8 SGs are connected to S.

[0119] In some embodiments of the vaccine, SG is independently selected from an amine, a hydroxyl, a carboxylic acid, and / or a sugar molecule, and the sugar molecule is independently selected from mannose, glucose, glucosamine, N-acetylglucose, galactose, galactosamine, N-acetylgalactosamine, N-acetylglucosamine, phosphoserine and any derivative thereof, an agonist of CD22a, sialyl Lewis x, and combinations thereof.

[0120] In some embodiments of the vaccine, the dendron amplifier comprises 1-10 generations of repeating monomer units with 2-6 branches per generation. In other embodiments, the dendron amplifier comprises 2-3 generations of repeating monomer units with 2-3 branches per generation. In some embodiments of the vaccine, the repeating monomer units are 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 is independently at each occurrence selected from: (CH2) y3 -FG2, (OCH2CH2) y3 -FG2 and CH2(OCH2CH2) y3 -FG2), where y2 and y3 are each, independently at each occurrence, an integer repeat unit of 1 to 6, FG1 is a first functional group, and FG2 is a second functional group. In some embodiments, FG1 is -NH2, and FG2 is, independently at each occurrence, -CO2- or -CO2H. In some embodiments, FG1 is, independently at each occurrence, -CO2- or -CO2H, and FG2 is -NH2.

[0121] In some embodiments of the vaccine, SG is linked to S via a suitable linker, X5. In some embodiments of the vaccine, a suitable linker, X5, linking SG to S is selected from lower alkyl and PEG groups. In some embodiments of the vaccine, two or more SGs are connected to the remainder of S by the dendron amplifier via a suitable linker, X5, which links two or more SGs to the terminal functional (FGt) group of the dendron amplifier via an amide bond. In some embodiments, the linker, X5, connecting SG to the dendron amplifier is -NH-R 19 , -NH-C(O)-R 19 , -C(O)-NH-R 19 -or-C(O)-R 19 wherein R 19 is, but is 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 -, where t and u are each independently an integer typically selected from 1 to 6, for example 1, 2, 3, 4, 5 or 6.

[0122] In some embodiments of the vaccine, the dendron amplifier comprises a polyethylene oxide (PEG) group.

[0123] In some embodiments of the vaccine, the amphiphile H comprises a higher alkane, an aromatic group, a fatty acid, a sterol, a polyunsaturated hydrocarbon, squalene, a saponin, and / or a polymer.

[0124] 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.

[0125] In some embodiments of the vaccine, each H independently comprises a poly(amino acid) comprising monomers selected from hydrophobic amino acids (M), reactive amino acids (N), spacer amino acids (O), charged amino acids (P), and combinations thereof, provided that at least one of M or N is present.

[0126] In some embodiments of the vaccine, each H independently has the formula: [ka] 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 of 1 to 100, and the sum of m, n, o, and p is 100 or less; R 3 are hydrogen, NH2, NH-CH3, NH-(CH2) y5 CH3, OH, or a drug molecule (D) connected either directly or via a suitable linker X1; y5 is an integer selected from 1 to 6.

[0127] In some embodiments of the vaccine, P is absent. In other embodiments, N, O, and P are each absent.

[0128] In some embodiments of the vaccine, P is [ka] where each R 5 are independently a group containing 1 to 2 charged functional groups.

[0129] In some embodiments of the vaccine, O is [ka] wherein each Q is independently (CH) y6 and (CH2CH2O) y7 CH2CH2, each y6 is independently selected from an integer of 1 to 6, and each y7 is independently selected from an integer of 1 to 4.

[0130] In some embodiments of the vaccine, N is [ka] 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 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 is included as an enzyme-degradable peptide and / or a self-immolative linker.

[0131] In some embodiments, X is present and -(CH) y10 -W and -(CH2) y10 -R 6 wherein y10 is an integer selected from 1 to 6; 6 is -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, wherein R 7 is -(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-CHR8 -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 each independently represent an integer selected from 1 to 6; 8is any amino acid side chain, and W can be independently selected from H (hydrogen), FG3, LG, and w, where FG3 can be selected from, but is not limited to, carboxylic acids, activated carboxylic acids (e.g., carbonylthiazolidine-2-thione ("TT"), NHS, or nitrophenol esters), carboxylic acid anhydrides, amines and protected amines (e.g., tert-butyloxycarbonyl-protected amines), OSi(CH3), alkenes, azides, alkynes, dye alkynes, halogens (e.g., fluorides, chlorides), olefins and endocyclic olefins (e.g., allyl), CN, OH, and epoxy, hydrazines (including hydrazides), carbohydrazides, aldehydes, ketones, carbamates, and activated carbamates. wherein w is any functional group suitable for attachment to a functional group present on a drug molecule ("FG4"), which may be selected from any suitable leaving group (e.g., NHS, TT, nitrophenol, etc.); w results from either the reaction of FG4 with FG3 or the displacement of LG with FG4 and is typically a group 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)-, where w is always linked to D either directly (i.e., w-D) or indirectly via X3 (i.e., w-X3-D).

[0132] In some embodiments of the vaccine, M is [ka] where each R 4 are independently hydrophobic groups.

[0133] In some embodiments of the vaccine, R 4 teeth, [ka] where: a is aryl or heteroaryl; X2 is present or absent and, if present, is a suitable linker; y8 is selected from integers from 0 to 6; Z 1 , Z 2 , and Z 3 are each independently selected from H, F, hydroxy, amino, alkyl, and fluoroalkyl.

[0134] In some embodiments of the vaccine, a is aryl, e.g., phenyl or naphthyl. In other embodiments, a is heteroaryl, e.g., pyridinyl, quinolinyl, isoquinolinyl, indolyl, or benzimidazolyl.

[0135] In some embodiments of the vaccine, X2 is absent. In other embodiments, X2 is present and is selected from the group consisting of C(O), CO2(CH2), y9 , CO2, C(O)NH(CH2) y9 , NHC(O) and NHC(O)(CH2) y9 where y9 is an integer typically selected from 1 to 6. In other embodiments, X2 is present and is selected from a lower alkyl and a PEG group.

[0136] In some embodiments of the vaccine, each R 4 is independent, [ka] wherein each X2 is independently selected from a suitable linker and each y8 is independently selected from an integer from 0 to 6. In other embodiments, each R 4 is independent, [ka] wherein each y is independently selected from an integer from 0 to 6. In other embodiments, each R 4 is independent, [ka] In other embodiments, each R 4 is independent, [ka] [ka] In a preferred embodiment, each R 4 is independent, [ka] is selected from.

[0137] In some embodiments of the vaccine, at least one D is [ka] where: R 20 is selected from H, alkyl, alkoxyalkyl, aryl, heteroaryl, aminoalkyl, amide and ester; X 3 is selected from alkyl, alkoxyalkyl, aralkyl, heteroaralkyl, aryl, heteroaryl, and carboxy.

[0138] In some embodiments of the vaccine, R 20 is selected from H, alkyl and alkoxyalkyl; X 3 is selected from alkyl and aralkyl. In other embodiments, R 20 is butyl.

[0139] In some embodiments of the vaccine, X 3 is alkyl.

[0140] In some embodiments of the vaccine, m, n, o, and p each independently represent an integer of 1 to 30, and the sum of m, n, o, and p is 30 or less.

[0141] 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 10 or less.

[0142] 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.

[0143] In some embodiments of the vaccine, the PEG group comprises 4 to 36 monomer units, while in other embodiments the PEG group comprises 4 to 12 monomer units.

[0144] In some embodiments of the vaccine, the hydrophilic peptide comprises 4 to 36 amino acids, hi other embodiments, the hydrophilic peptide comprises 4 to 12 amino acids.

[0145] 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.

[0146] In some embodiments of the vaccine, the vaccine comprises a molar ratio of peptide antigen conjugate to amphiphile of about 4:1 to about 1:20, preferably about 1:1.

[0147] 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.

[0148] In some embodiments of the vaccine, the peptide antigen (A) comprises a sequence in which one or more cysteine ​​residues are replaced with alpha-aminobutyric acid and / or one or more methionine residues are replaced with 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, infectious disease, and tolerance-inducing vaccines, as well as for the treatment of cardiovascular disease or cardiovascular disease.

[0149] In some embodiments of the vaccine, A is selected from the group consisting of RGYLTKILHVFHGLLPGFLVKMSGDLLE (SEQ ID NO: 52), PGFLVKMSGDLLE (SEQ ID NO: 53), PGFLVKnSGDLLE (where n = norleucine) (SEQ ID NO: 54), PGFLVKMSSDLLG (SEQ ID NO: 55), PGFLVKnSSDLLG (where n = norleucine) (SEQ ID NO: 56), SIPWNLERITPPR (SEQ ID NO: 57), SIPWNLERITPPR (SEQ ID NO: 58), SIPWN LE (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), NVPEEDG (SEQ ID NO: 65), NVPEEDATRFHRQGSK (SEQ ID NO: 66), LFAPGEDIIGASSDCSTCFVSQSGTSQAAA (SEQ ID NO: 67), CSTCFVSQSGTSQAAA (SEQ ID NO: 68), ST CFVSQSGTSQAAA (SEQ ID NO: 69), STBFVSQSGTSQAAA (SEQ ID NO: 70), STBFVSQ (SEQ ID NO: 71), MFTIKLLLFIVPLVISSRIDQDNSSFDSLSPEPKSRFAMLDDVKILANGLLQLGHGLKDFVHKTKGQIND (SEQ ID NO: 72), EPKSRFAMLDDVKILANGLLQLGHGLKDFVHKTKGQIND (SEQ ID NO: 73), EPKSRFAMLDDVKI (SEQ ID NO: 74), MLDDVKILANGLLQ (SEQ ID NO: 75), No. 75), LANGLLQLGHGLKD (SEQ ID NO: 76), LGHGLKDFVHKTKG (SEQ ID NO: 77), LKDFVHKTKGQIND (SEQ ID NO: 78), RFAMLDDVKILANGLLQLGH (SEQ ID NO: 79), GLLQLGHGLKDFVHKTKGQI (SEQ ID NO: 80), and IFQKLNIFDQSFYDLSLQTSEIKEEEKELRRTTYKLQVKNEEVKNMSLELNSKLESLLEEKILLQQKVK (SEQ ID NO: 81).

[0150] In some embodiments of the vaccine, A is covalently linked directly to E1, which is covalently linked directly to H or indirectly via U.

[0151] In some embodiments of the vaccine, A is covalently linked directly to E2, which is covalently linked directly to H or indirectly via U.

[0152] In some embodiments of the vaccine, E1 and E2 each comprise a PEG group of 4 to 36 monomer units, for example, a PEG group comprising 4 to 24 monomer units.

[0153] In some embodiments of the vaccine, E1 and E2 each comprise a peptide.

[0154] In some embodiments of the vaccine, the peptide comprises between 4 and 24 amino acids.

[0155] In some embodiments of the vaccine, at least one peptide antigen conjugate comprises A selected from an autoantigen, an alloantigen, and an allergen.

[0156] In some embodiments of the vaccine, the amphiphile, S, comprises two or more solubilizing groups (SG) independently selected from a carboxylic acid, a phosphoserine, and / or a sugar molecule, wherein the sugar molecule is independently selected from mannose, glucose, glucosamine, N-acetylglucose, galactose, galactosamine, and N-acetylgalactosamine, and an agonist of CD22a.

[0157] In some embodiments of the vaccine, the vaccine comprises an inhibitor of mTOR, RORgt, CDK8 / 19, and HDAC, as well as an inhibitor of AHR, RAR, and A 2a In some embodiments of the vaccine, the at least one D is selected from an ATP-competitive mTOR inhibitor.

[0158] In some embodiments of the vaccine, the vaccine comprises an inhibitor of mTOR, RORgt, CDK8 / 19, and HDAC, AHR, RAR, and A 2a and an immunostimulant selected from agonists of NLR, CLR, TLR and STING, provided that D and D2 bind to different receptors.

[0159] In some embodiments of the vaccine, at least one D is selected from an inhibitor of mTOR and an agonist of AHR, and D2 is selected from an agonist of NLR, CLR, TLR, and STING. In some embodiments of the vaccine, at least one D is selected from an ATP-competitive mTOR inhibitor, and D2 is selected from an agonist of NLR, CLR, TLR, and STING.

[0160] In some embodiments of the vaccine, D2 is selected from an agonist of TLR-3, TLR-7, TLR-8, TLR-7 / 8, TLR-9, and STING. In some embodiments of the vaccine, D2 is selected from RNA and an imidazoquinoline agonist of TLR-7, TLR-8, and TLR-7 / 8.

[0161] In some embodiments of the vaccine, the vaccine comprises an inhibitor of mTOR, RORgt, CDK8 / 19, and HDAC, AHR, RAR, and A 2a and an immunostimulant selected from agonists of NLR, CLR, TLR and STING, provided that D, D2 and D3 bind to different receptors.

[0162] In some embodiments of the vaccine, 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.

[0163] In some embodiments of the vaccine, the molar ratio of total peptide antigen conjugate to at least one D is about 20:1 to 1:2, or about 10:1 to about 1:1, or about 4:1 to about 2:1, preferably about 1:1.

[0164] In some embodiments of the vaccine, at least one peptide antigen conjugate comprises A selected from a tumor antigen.

[0165] In some embodiments of the vaccine, the amphiphile S comprises two or more solubilizing groups (SG) independently selected from an amine or a sugar molecule, wherein the sugar molecule is independently selected from mannose and sialyl Lewis x, and combinations thereof. In some embodiments of the vaccine, the amphiphile S comprises two or more solubilizing groups (SG) independently selected from an amine, a carboxylic acid, or a sugar molecule, wherein the sugar molecule is 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.

[0166] In some embodiments of the vaccine, each H of the amphiphile and / or peptide antigen conjugate independently comprises a poly(amino acid) comprising monomers of a hydrophobic amino acid (M) selected from tryptophan, 1-methyltryptophan, and para-aminophenylalanine. In other embodiments, each H of the amphiphile and / or peptide antigen conjugate comprises a poly(amino acid) comprising monomers of a reactive amino acid (N), wherein the monomers comprise a D selected from Glu-TLR-7 / 8a. In some embodiments of the vaccine, at least one D is present and is 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 peptide antigen conjugate to D2 is about 20:1 to 1:2, or about 10:1 to about 1:1, or about 4:1 to about 2:1, preferably about 1:1.

[0167] In some embodiments of the vaccine, A is a glycopeptide. In other embodiments, A is a glycopeptide. * 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), wherein: * is an O-linked glycan, and each occurrence is independently selected from sialyl Lewis x, sialyl Lewis a, Lewis y, Lewis x, Tn, sTn, TF, and sTF.

[0168] In some embodiments of the vaccine, the amphiphile S comprises a second or third generation dendrimer and B comprises 4 to 36 PEG monomer units.

[0169] In some embodiments of the vaccine, the amphiphile S comprises a second or third generation dendrimer and the amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0170] In some embodiments of the vaccine, B comprises 4 to 36 monomer units and the amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0171] In some embodiments of the vaccine, amphiphile S comprises a second or third generation dendrimer, B comprises 4 to 36 PEG monomer units, and amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0172] In some embodiments of the vaccine, amphiphile S comprises a second or third generation dendrimer and amphiphile H comprises a poly(amino acid) comprising para-amino-phenylalanine.

[0173] In some embodiments of the vaccine, B comprises 4 to 36 PEG monomer units and the amphiphile H comprises a polymer of para-amino-phenylalanine.

[0174] In some embodiments of the vaccine, amphiphile S comprises a second or third generation dendrimer, B comprises 4 to 36 PEG monomer units, and amphiphile H comprises a polymer of para-amino-phenylalanine.

[0175] In some embodiments of the vaccine, the amphiphile S comprises a second or third generation dendrimer and the amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and a reactive amino acid (N) comprising an imidazoquinoline.

[0176] In some embodiments of the vaccine, B comprises 4 to 36 PEG monomer units, and the amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and a reactive amino acid (N) including an imidazoquinoline.

[0177] In some embodiments of the vaccine, amphiphile S comprises a second or third generation dendrimer, B comprises 4 to 36 PEG monomer units, and amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and a reactive amino acid (N) including an imidazoquinoline.

[0178] In some embodiments of the vaccine, the amphiphile S comprises a second or third generation dendrimer and the amphiphile H comprises a poly(amino acid) of reactive amino acids (N) including tryptophan and imidazoquinoline.

[0179] In some embodiments of the vaccine, B comprises 4 to 36 PEG monomer units and the amphiphile H comprises a poly(amino acid) of reactive amino acids (N) including tryptophan and imidazoquinoline.

[0180] In some embodiments of the vaccine, amphiphile S comprises a second or third generation dendrimer, B comprises 4 to 36 PEG monomer units, and amphiphile H comprises a poly(amino acid) of reactive amino acids (N) including tryptophan and imidazoquinoline.

[0181] In some embodiments of the vaccine, amphiphile S comprises a second or third generation dendrimer, B comprises 4 to 36 PEG monomer units, and SG comprises mannose.

[0182] In some embodiments of the vaccine, the amphiphile S comprises a second or third generation dendrimer, the amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M), and the SG comprises mannose.

[0183] In some embodiments of the vaccine, B comprises 4 to 36 PEG monomer units, the amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M), and SG comprises mannose.

[0184] In some embodiments of the vaccine, amphiphile S comprises a second or third generation dendrimer, B comprises 4 to 36 PEG monomer units, amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M), and SG comprises mannose.

[0185] In some embodiments of the vaccine, amphiphile S comprises a second or third generation dendrimer, amphiphile H comprises a polymer of para-amino-phenylalanine, and SG comprises mannose.

[0186] In some embodiments of the vaccine, B comprises 4 to 36 PEG monomer units, the amphiphile H comprises a polymer of para-amino-phenylalanine, and SG comprises mannose.

[0187] In some embodiments of the vaccine, amphiphile S comprises a second or third generation dendrimer, B comprises 4 to 36 PEG monomer units, amphiphile H comprises a polymer of para-amino-phenylalanine, and SG comprises mannose.

[0188] In some embodiments of the vaccine, the amphiphile S comprises a second or third generation dendrimer, the amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and a reactive amino acid (N) comprising an imidazoquinoline, and the amphiphile SG comprises mannose.

[0189] In some embodiments of the vaccine, B comprises 4 to 36 PEG monomer units, the amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and a reactive amino acid (N) comprising an imidazoquinoline, and SG comprises mannose.

[0190] In some embodiments of the vaccine, amphiphile S comprises a second or third generation dendrimer, B comprises 4 to 36 PEG monomer units, amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and a reactive amino acid (N) comprising an imidazoquinoline, and SG comprises mannose.

[0191] In some embodiments of the vaccine, the amphiphile S comprises a second or third generation dendrimer, the amphiphile H comprises a poly(amino acid) of reactive amino acids (N) including tryptophan and imidazoquinoline, and the amphiphile SG comprises mannose.

[0192] In some embodiments of the vaccine, B comprises 4 to 36 PEG monomer units, the amphiphile H comprises a poly(amino acid) of reactive amino acids (N) including tryptophan and imidazoquinoline, and SG comprises mannose.

[0193] In some embodiments of the vaccine, amphiphile S comprises a second or third generation dendrimer, B comprises 4 to 36 PEG monomer units, amphiphile H comprises a poly(amino acid) of reactive amino acids (N) including tryptophan and imidazoquinoline, and SG comprises mannose.

[0194] In some embodiments of the vaccine, the amphiphile S comprises a second or third generation dendrimer, B comprises 4 to 36 PEG monomer units, and the peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0195] In some embodiments of the vaccine, the amphiphile S comprises a second or third generation dendrimer, the amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M), and the peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0196] 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).

[0197] In some embodiments of the vaccine, amphiphile S comprises a second or third generation dendrimer, B comprises 4 to 36 PEG monomer units, amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M), and peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0198] In some embodiments of the vaccine, the amphiphile S comprises a second or third generation dendrimer, the amphiphile H comprises a polymer of para-amino-phenylalanine, and the peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0199] In some embodiments of the vaccine, B comprises 4 to 36 PEG monomer units, the amphiphile H comprises a polymer of para-amino-phenylalanine, and the peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0200] In some embodiments of the vaccine, amphiphile S comprises a second or third generation dendrimer, B comprises 4 to 36 PEG monomer units, amphiphile H comprises a polymer of para-amino-phenylalanine, and peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0201] In some embodiments of the vaccine, the amphiphile S comprises a second or third generation dendrimer, the amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and a reactive amino acid (N) comprising an imidazoquinoline, and the peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0202] 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) comprising an imidazoquinoline, and H of the peptide antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0203] In some embodiments of the vaccine, amphiphile S comprises a second or third generation dendrimer, B comprises 4 to 36 PEG monomer units, amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and a reactive amino acid (N) comprising an imidazoquinoline, and peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0204] In some embodiments of the vaccine, the amphiphile S comprises a second or third generation dendrimer, the amphiphile H comprises a poly(amino acid) of reactive amino acids (N) including tryptophan and imidazoquinoline, and the peptide antigen conjugate H comprises a poly(amino acid) including a hydrophobic amino acid (M).

[0205] In some embodiments of the vaccine, B comprises 4-36 PEG monomer units, the amphiphile H comprises a poly(amino acid) of reactive amino acids (N) including tryptophan and imidazoquinoline, and the peptide antigen conjugate H comprises a poly(amino acid) including a hydrophobic amino acid (M).

[0206] In some embodiments of the vaccine, amphiphile S comprises a second or third generation dendrimer, B comprises 4 to 36 PEG monomer units, amphiphile H comprises a poly(amino acid) of reactive amino acids (N) including tryptophan and imidazoquinoline, and peptide antigen conjugate H comprises a poly(amino acid) including a hydrophobic amino acid (M).

[0207] In some embodiments of the vaccine, the amphiphile S 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) comprising a hydrophobic amino acid (M).

[0208] In some embodiments of the vaccine, the amphiphile S comprises a second or third generation dendrimer, the amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M), the SG comprises mannose, and the peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0209] In some embodiments of the vaccine, B comprises 4 to 36 PEG monomer units, the amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M), the SG comprises mannose, and the peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0210] In some embodiments of the vaccine, amphiphile S comprises a second or third generation dendrimer, B comprises 4 to 36 PEG monomer units, amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M), SG comprises mannose, and peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0211] In some embodiments of the vaccine, the amphiphile S comprises a second or third generation dendrimer, the amphiphile H comprises a polymer of para-amino-phenylalanine, the SG comprises mannose, and the peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0212] In some embodiments of the vaccine, B comprises 4 to 36 PEG monomer units, the amphiphile H comprises a polymer of para-amino-phenylalanine, SG comprises mannose, and the peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0213] In some embodiments of the vaccine, amphiphile S comprises a second or third generation dendrimer, B comprises 4 to 36 PEG monomer units, amphiphile H comprises a polymer of para-amino-phenylalanine, SG comprises mannose, and peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0214] In some embodiments of the vaccine, amphiphile S comprises a second or third generation dendrimer, amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and a reactive amino acid (N) comprising an imidazoquinoline, SG comprises mannose, and peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0215] 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) including an imidazoquinoline, SG comprises mannose, and H of the peptide antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0216] In some embodiments of the vaccine, amphiphile S comprises a second or third generation dendrimer, B comprises 4 to 36 PEG monomer units, amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and a reactive amino acid (N) including an imidazoquinoline, SG comprises mannose, and peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0217] In some embodiments of the vaccine, the amphiphile S comprises a second or third generation dendrimer, the amphiphile H comprises a poly(amino acid) of reactive amino acids (N) including tryptophan and imidazoquinoline, the SG comprises mannose, and the peptide antigen conjugate H comprises a poly(amino acid) including a hydrophobic amino acid (M).

[0218] In some embodiments of the vaccine, B comprises 4-36 PEG monomer units, the amphiphile H comprises a poly(amino acid) of reactive amino acids (N) including tryptophan and imidazoquinoline, SG comprises mannose, and the peptide antigen conjugate H comprises a poly(amino acid) including a hydrophobic amino acid (M).

[0219] In some embodiments of the vaccine, amphiphile S comprises a second or third generation dendrimer, B comprises 4 to 36 PEG monomer units, amphiphile H comprises a poly(amino acid) of reactive amino acids (N) including tryptophan and imidazoquinoline, SG comprises mannose, and peptide antigen conjugate H comprises a poly(amino acid) including a hydrophobic amino acid (M).

[0220] In some embodiments of the vaccine, the amphiphile S 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) comprising a hydrophobic amino acid (M).

[0221] In some embodiments of the vaccine, the amphiphile S comprises a second or third generation dendrimer, the amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M), the amphiphile comprises amino-hexanoic acid, and the peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0222] 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).

[0223] In some embodiments of the vaccine, amphiphile S comprises a second or third generation dendrimer, B comprises 4 to 36 PEG monomer units, amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M), the amphiphile comprises amino-hexanoic acid, and peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0224] In some embodiments of the vaccine, the amphiphile S comprises a second or third generation dendrimer, the amphiphile H comprises a polymer of para-amino-phenylalanine, the amphiphile comprises amino-hexanoic acid, and the peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0225] In some embodiments of the vaccine, B comprises 4 to 36 PEG monomer units, the amphiphile H comprises a polymer of para-amino-phenylalanine, the amphiphile comprises amino-hexanoic acid, and the peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0226] In some embodiments of the vaccine, the amphiphile S comprises a second or third generation dendrimer, B comprises 4 to 36 PEG monomer units, the amphiphile H comprises a polymer of para-amino-phenylalanine, the amphiphile comprises amino-hexanoic acid, and the peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0227] 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 a reactive amino acid (N) comprising an imidazoquinoline, the amphiphile comprises an amino-hexanoic acid, and H of the peptide antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0228] 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) comprising an imidazoquinoline, the amphiphile comprises an amino-hexanoic acid, and H of the peptide antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0229] In some embodiments of the vaccine, amphiphile S comprises a second or third generation dendrimer, B comprises 4 to 36 PEG monomer units, amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and a reactive amino acid (N) including an imidazoquinoline, the amphiphile comprises amino-hexanoic acid, and peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0230] In some embodiments of the vaccine, the amphiphile S comprises a second or third generation dendrimer, the amphiphile H comprises a poly(amino acid) of reactive amino acids (N) including tryptophan and imidazoquinoline, the amphiphile comprises amino-hexanoic acid, and the peptide antigen conjugate H comprises a poly(amino acid) including a hydrophobic amino acid (M).

[0231] In some embodiments of the vaccine, B comprises 4-36 PEG monomer units, the amphiphile H comprises a poly(amino acid) of reactive amino acids (N) including tryptophan and imidazoquinoline, the amphiphile comprises amino-hexanoic acid, and the peptide antigen conjugate H comprises a poly(amino acid) including a hydrophobic amino acid (M).

[0232] In some embodiments of the vaccine, amphiphile S comprises a second or third generation dendrimer, B comprises 4 to 36 PEG monomer units, amphiphile H comprises a poly(amino acid) of reactive amino acids (N) including tryptophan and imidazoquinoline, the amphiphile comprises amino-hexanoic acid, and peptide antigen conjugate H comprises a poly(amino acid) including a hydrophobic amino acid (M).

[0233] In some embodiments of the vaccine, the amphiphile S comprises a second or third generation dendrimer, B comprises 4 to 36 PEG monomer units, 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).

[0234] In some embodiments of the vaccine, the amphiphile S comprises a second or third generation dendrimer, the amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M), the dendrimer monomer comprises a hydroxy acid and an amino alcohol, and the peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0235] In some embodiments of the vaccine, B comprises 4-36 PEG monomer units, the amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M), the dendrimer monomer comprises a hydroxy acid and an amino alcohol, and the peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0236] In some embodiments of the vaccine, the amphiphile S comprises a second or third generation dendrimer, B comprises 4 to 36 PEG monomer units, the amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M), the dendrimer monomer comprises a hydroxy acid and an amino alcohol, and the peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0237] In some embodiments of the vaccine, the amphiphile S comprises a second or third generation dendrimer, the amphiphile H comprises a polymer of para-amino-phenylalanine, the dendrimer monomer comprises a hydroxy acid and an amino alcohol, and the peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0238] In some embodiments of the vaccine, B comprises 4-36 PEG monomer units, the amphiphile H comprises a polymer of para-amino-phenylalanine, the dendrimer monomer comprises a hydroxy acid and an amino alcohol, and the peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0239] In some embodiments of the vaccine, amphiphile S comprises a second or third generation dendrimer, B comprises 4 to 36 PEG monomer units, amphiphile H comprises a polymer of para-amino-phenylalanine, the dendrimer monomer comprises a hydroxy acid and an amino alcohol, and peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0240] In some embodiments of the vaccine, the amphiphile S comprises a second or third generation dendrimer, the amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and a reactive amino acid (N) including an imidazoquinoline, the dendrimer monomer comprises a hydroxy acid and an amino alcohol, and the peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0241] In some embodiments of the vaccine, B comprises 4-36 PEG monomer units, the amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and a reactive amino acid (N) including an imidazoquinoline, the dendrimer monomer comprises a hydroxy acid and an amino alcohol, and the peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0242] In some embodiments of the vaccine, amphiphile S comprises a second or third generation dendrimer, B comprises 4 to 36 PEG monomer units, amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and a reactive amino acid (N) including an imidazoquinoline, the dendrimer monomer comprises a hydroxy acid and an amino alcohol, and peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0243] In some embodiments of the vaccine, the amphiphile S comprises a second or third generation dendrimer, the amphiphile H comprises a poly(amino acid) of reactive amino acids (N) including tryptophan and imidazoquinoline, the dendrimer monomer comprises a hydroxy acid and an amino alcohol, and the peptide antigen conjugate H comprises a poly(amino acid) including a hydrophobic amino acid (M).

[0244] In some embodiments of the vaccine, B comprises 4-36 PEG monomer units, the amphiphile H comprises a poly(amino acid) of reactive amino acids (N) including tryptophan and imidazoquinoline, the dendrimer monomer comprises a hydroxy acid and an amino alcohol, and the peptide antigen conjugate H comprises a poly(amino acid) including a hydrophobic amino acid (M).

[0245] In some embodiments of the vaccine, amphiphile S comprises a second or third generation dendrimer, B comprises 4 to 36 PEG monomer units, amphiphile H comprises a poly(amino acid) of reactive amino acids (N) including tryptophan and imidazoquinoline, dendrimer monomers comprise hydroxy acids and amino alcohols, and peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0246] The present disclosure relates to a vaccine for inducing tolerance 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, During the ceremony, A is a peptide antigen, E1 is the N-terminal extension, E2 is a C-terminal extension, H, independently at each occurrence, is a hydrophobic block; and one or more drug molecules (D) are optionally attached to each H, either directly or via a suitable linker, X1; U is, independently at each occurrence, a linker; [ ] means that the group is optional; - denotes that two adjacent groups are directly linked to each other by a covalent bond or indirectly linked to each other via a suitable linker X, wherein at least one peptide antigen is selected from an autoantigen, an alloantigen and an allergen.

[0247] In some embodiments of the vaccine, an amphiphile is present when either (i) the average water 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.

[0248] The present disclosure also relates to a vaccine for inducing tolerance 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, During the ceremony, A is a peptide antigen, E1 is the N-terminal extension, E2 is a C-terminal extension, H, independently at each occurrence, is a hydrophobic block; and one or more drug molecules (D) are optionally attached to each H, either directly or via a suitable linker, X1; S is a solubilizing block, B is a spacer, U is, independently at each occurrence, a linker; [ ] means that the group is optional; - denotes that two adjacent groups are directly linked to each other by a covalent bond or indirectly linked to each other via a suitable linker X; wherein the amphiphilic material comprises a dendron amplifier material and the at least one peptide antigen is selected from an autoantigen, an alloantigen, and an allergen.

[0249] In one embodiment of the vaccine, the PEG group of the peptide antigen conjugate comprises a terminal functional group selected from OH, MeO-, and NH2.

[0250] In a particular embodiment of the vaccine for inducing tolerance, the PEG group of the peptide antigen conjugate is polyethylene glycol.

[0251] In some embodiments of the vaccine for inducing tolerance, the PEG group of the peptide antigen conjugate comprises between 4 and 36 monomer units.

[0252] In certain embodiments of the vaccine for inducing tolerance, the PEG group of the peptide antigen conjugate comprises 4 to 12 monomer units, or 12 to 36 monomer units, preferably 24 monomer units.

[0253] In some embodiments of the vaccine for inducing tolerance, A is an autoantigen. In other embodiments, A is an allergen. In other embodiments, A is an alloantigen.

[0254] In some embodiments of the vaccine for inducing tolerance, when an amphiphile is present, the amphiphile comprises a dendron amplifier.

[0255] In some embodiments of the vaccine for inducing tolerance, the amphiphile S comprises a dendron amplifier. In other embodiments, the amphiphile S has a dendritic structure.

[0256] In some embodiments of vaccines for inducing tolerance, the amphiphile S comprises two or more solubilizing groups (SGs). In other embodiments, two or more SGs are connected to the remainder of the S by a dendron amplifier, e.g., 4 to 8 SGs are connected to the S.

[0257] In some embodiments of the vaccine for inducing tolerance, SG is independently selected from an amine, a hydroxyl, a carboxylic acid, and / or a sugar molecule, wherein the sugar molecule is independently selected from mannose, glucose, glucosamine, N-acetylglucose, galactose, galactosamine, N-acetylgalactosamine, N-acetylglucosamine, phosphoserine and any derivative thereof, an agonist of CD22a, sialyl Lewis x, and combinations thereof.

[0258] 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 a CD22a agonist.

[0259] In some embodiments of the vaccine for inducing tolerance, the dendron amplifier comprises 1 to 10 generations of repeating monomer units with 2 to 6 branches per generation, while in other embodiments, the dendron amplifier comprises 2 to 3 generations of repeating monomer units with 2 to 3 branches per generation.

[0260] In some embodiments of the vaccine for inducing tolerance, the repeating monomer unit is 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 R 1 is independently calculated for each occurrence, (CH2) y3 -FG2, (OCH2CH2) y3 -FG2 and CH2(OCH2CH2) y3-FG2), where y2 and y3 are, independently at each occurrence, an integer repeat unit of 1 to 6, FG1 is a first functional group, and FG2 is a second functional group. In some embodiments, FG1 is -NH2, and FG2 is, independently at each occurrence, -CO2- or -CO2H. In some embodiments, FG1 is -CO2- or -CO2H, and FG2 is, independently at each occurrence, -NH2.

[0261] 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 lower alkyl and PEG groups. In some embodiments of the vaccine for inducing tolerance, two or more SGs are connected to the remainder of S by the dendron amplifier via a suitable linker, X5, which links two or more SGs to the terminal functional (FGt) group of the dendron amplifier by an amide bond. In some embodiments of the vaccine for inducing tolerance, the linker X5 joining SG to the dendron amplifier is -NH-R 19 , -NH-C(O)-R 19 , -C(O)-NH-R 19 -or-C(O)-R 19 wherein R 19 is, but is 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 -, where t and u are each independently an integer typically selected from 1 to 6, for example 1, 2, 3, 4, 5 or 6.

[0262] In some embodiments of the vaccine for inducing tolerance, the dendron amplifier comprises a polyethylene oxide (PEG) group.

[0263] In some embodiments of the vaccine for inducing tolerance, the amphiphile H comprises a higher alkane, an aromatic group, a fatty acid, a sterol, a polyunsaturated hydrocarbon, squalene, a saponin, and / or a polymer.

[0264] In some embodiments of a 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.

[0265] In some embodiments of a vaccine for inducing tolerance, each H independently comprises a poly(amino acid) comprising monomers selected from hydrophobic amino acids (M), reactive amino acids (N), spacer amino acids (O), charged amino acids (P), and combinations thereof, provided that at least one of M or N is present.

[0266] In some embodiments of the vaccine for inducing tolerance, each H independently has the formula: [ka] 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 of 1 to 100, and the sum of m, n, o, and p is 100 or less; R 3 are hydrogen, NH2, NH-CH3, NH-(CH2) y5 CH3, OH, or a drug molecule (D) connected either directly or via a suitable linker X1; y5 is an integer selected from 1 to 6.

[0267] In some embodiments of the vaccine for inducing tolerance, P is absent. In other embodiments, N, O, and P are each absent.

[0268] In some embodiments of the vaccine for inducing tolerance, P is [ka] where each R 5 are independently a group containing 1 to 2 charged functional groups.

[0269] In some embodiments of the vaccine for inducing tolerance, O is [ka] wherein each Q is independently (CH) y and (CH2CH2O) i CH2CH2, each y is independently selected from an integer of 1 to 6, and each i is independently selected from an integer of 1 to 4.

[0270] In some embodiments of the vaccine for inducing tolerance, N is [ka] wherein each X1 is independently a suitable linker and each D is independently a drug molecule.

[0271] In some embodiments of the vaccine for inducing tolerance, M is [ka] where each R 4 are independently hydrophobic groups.

[0272] In some embodiments of the vaccine for inducing tolerance, R 4 teeth, [ka] where: a is aryl or heteroaryl; X2 is present or absent and, if present, is a suitable linker; Y8 is selected from integers from 0 to 6; Z 1 , Z 2 , and Z 3 are each independently selected from H, F, hydroxy, amino, alkyl, and fluoroalkyl.

[0273] In some embodiments of the vaccine for inducing tolerance, a is aryl, e.g., phenyl or naphthyl. In other embodiments, a is heteroaryl, e.g., pyridinyl, quinolinyl, isoquinolinyl, indolyl, or benzimidazolyl.

[0274] In some embodiments of the vaccine for inducing tolerance, X is absent. In other embodiments, X is present and is selected from the group consisting of C(O), CO(CH) y9 , CO2, C(O)NH(CH2) y9 , NHC(O) and NHC(O)(CH2) y9 where y9 is an integer typically selected from 1 to 6. In other embodiments, X2 is present and is selected from a lower alkyl and a PEG group.

[0275] In some embodiments of the vaccine for inducing tolerance, each R 4 is independent, [ka] wherein each X2 is independently selected from a suitable linker and each y8 is independently selected from an integer from 0 to 6. In other embodiments, each R 4 is independent, [ka] wherein each y is independently selected from an integer from 0 to 6. In other embodiments, each R 4is independent, [ka] In other embodiments, each R 4 is independent, [ka] [ka] In a preferred embodiment, each R 4 is independent, [ka] wherein y is selected from integers of 1 to 6.

[0276] In some embodiments of the vaccine for inducing tolerance, at least one D is [ka] where: R 20 is selected from H, alkyl, alkoxyalkyl, aryl, heteroaryl, aminoalkyl, amide, and ester; and X3 is selected from alkyl, alkoxyalkyl, aralkyl, heteroaralkyl, aryl, heteroaryl, and carboxy.

[0277] In some embodiments of the vaccine for inducing tolerance, R 20 is selected from H, alkyl and alkoxyalkyl, and X3 is selected from alkyl and aralkyl. 20 is butyl.

[0278] In some embodiments of the vaccine for inducing tolerance, X3 is alkyl.

[0279] In some embodiments of the vaccine for inducing tolerance, m, n, o, and p each independently represent an integer of 1 to 30, and the sum of m, n, o, and p is 30 or less.

[0280] In some embodiments of the vaccine for inducing tolerance, m, n, o, and p each independently represent an integer of 1 to 10, and the sum of m, n, o, and p is 10 or less.

[0281] In some embodiments of the vaccine 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.

[0282] In some embodiments of the vaccine for inducing tolerance, the PEG group comprises 4 to 36 monomer units, while in other embodiments, the PEG group comprises 4 to 12 monomer units.

[0283] In some embodiments of the vaccine for inducing tolerance, the hydrophilic peptide comprises 4 to 36 amino acids, hi other embodiments, the hydrophilic peptide comprises 4 to 12 amino acids.

[0284] In some embodiments of the vaccine 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.

[0285] In some embodiments of a vaccine for inducing tolerance, the vaccine comprises a molar ratio of peptide antigen conjugate to amphiphile of about 4:1 to about 1:20, preferably about 1:1.

[0286] In some embodiments of the vaccine for inducing tolerance, the peptide antigen (A) comprises a sequence in which one or more cysteine ​​residues are replaced with alpha-aminobutyric acid and / or one or more methionine residues are replaced with norleucine.

[0287] In some embodiments of the vaccine for inducing tolerance, the peptide antigen (A) comprises alpha aminobutyric acid and / or norleucine.

[0288] In some embodiments of a vaccine for inducing tolerance, the vaccine comprises an inhibitor of mTOR, RORgt, CDK8 / 19, and HDAC, as well as an inhibitor of AHR, RAR, and A. 2a In other embodiments, the at least one D is selected from an ATP-competitive mTOR inhibitor.

[0289] In some embodiments of a vaccine for inducing tolerance, the vaccine comprises an inhibitor of mTOR, RORgt, CDK8 / 19, and HDAC, AHR, RAR, and A 2a and an immunostimulant selected from agonists of NLR, CLR, TLR and STING, provided that D and D2 bind to different receptors.

[0290] In some embodiments of the vaccine for inducing tolerance, at least one D is selected from an inhibitor of mTOR and an agonist of AHR, and D2 is selected from an agonist of NLR, CLR, TLR, and STING.

[0291] In some embodiments of the vaccine for inducing tolerance, at least one D is selected from an ATP-competitive mTOR inhibitor and D2 is selected from an agonist of an NLR, a CLR, a TLR, and a STING.

[0292] In other embodiments of the vaccine for inducing tolerance, D2 is selected from an agonist of TLR-3, TLR-7, TLR-8, TLR-7 / 8, TLR-9, and STING. In other embodiments of the vaccine for inducing tolerance, D2 is selected from RNA and an imidazoquinoline agonist of TLR-7, TLR-8, and TLR-7 / 8.

[0293] In some embodiments of a vaccine for inducing tolerance, the vaccine comprises an inhibitor of mTOR, RORgt, CDK8 / 19, and HDAC, AHR, RAR, and A 2a and an immunostimulant selected from agonists of NLR, CLR, TLR and STING, provided that D, D2 and D3 bind to different receptors.

[0294] In some embodiments of a vaccine for inducing tolerance, 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.

[0295] In some embodiments of a vaccine for inducing tolerance, the molar ratio of total peptide antigen conjugate to at least one D is about 20:1 to 1:2, or about 10:1 to about 1:1, or about 4:1 to about 2:1, or preferably about 1:1.

[0296] In some embodiments of the vaccine for inducing tolerance, the amphiphile S comprises a second or third generation dendrimer and B comprises 4 to 36 PEG monomer units.

[0297] In some embodiments of the vaccine for inducing tolerance, the amphiphile S comprises a second or third generation dendrimer and the amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0298] In some embodiments of the vaccine for inducing tolerance, B comprises 4 to 36 PEG monomer units and the amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0299] In some embodiments of a vaccine for inducing tolerance, amphiphile S comprises a second or third generation dendrimer, amphiphile B comprises 4 to 36 PEG monomer units, and amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0300] In some embodiments of the vaccine for inducing tolerance, the amphiphile S comprises a second or third generation dendrimer and the amphiphile H comprises a polymer of para-amino-phenylalanine.

[0301] In some embodiments of the vaccine for inducing tolerance, B comprises 4 to 36 PEG monomer units and the amphiphile H comprises a polymer of para-amino-phenylalanine.

[0302] In some embodiments of the vaccine for inducing tolerance, amphiphile S comprises a second or third generation dendrimer, amphiphile B comprises 4 to 36 PEG monomer units, and amphiphile H comprises a polymer of para-amino-phenylalanine.

[0303] In some embodiments of a vaccine for inducing tolerance, the amphiphile S comprises a second or third generation dendrimer, and the amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and a reactive amino acid (N) comprising an imidazoquinoline.

[0304] In some embodiments of the vaccine for inducing tolerance, B comprises 4 to 36 PEG monomer units, and the amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and a reactive amino acid (N) including an imidazoquinoline.

[0305] In some embodiments of a vaccine for inducing tolerance, amphiphile S comprises a second or third generation dendrimer, amphiphile B comprises 4 to 36 PEG monomer units, and amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and a reactive amino acid (N) including an imidazoquinoline.

[0306] In some embodiments of the vaccine for inducing tolerance, the amphiphile S comprises a second or third generation dendrimer and the amphiphile H comprises a poly(amino acid) of reactive amino acids (N) including tryptophan and imidazoquinoline.

[0307] In some embodiments of the vaccine for inducing tolerance, B comprises 4 to 36 PEG monomer units, and the amphiphile H comprises a poly(amino acid) of reactive amino acids (N) including tryptophan and imidazoquinoline.

[0308] In some embodiments of the vaccine for inducing tolerance, the amphiphile S comprises a second or third generation dendrimer, the amphiphile B comprises 4 to 36 PEG monomer units, and the amphiphile H comprises a poly(amino acid) of reactive amino acids (N) including tryptophan and imidazoquinoline.

[0309] In some embodiments of the vaccine for inducing tolerance, the amphiphile S comprises a second or third generation dendrimer, B comprises 4 to 36 PEG monomer units, and SG comprises N-acetylgalactosamine.

[0310] In some embodiments of a vaccine for inducing tolerance, the amphiphile S comprises a second or third generation dendrimer, the amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M), and the amphiphile SG comprises N-acetylgalactosamine.

[0311] In some embodiments of the vaccine for inducing tolerance, B comprises 4 to 36 PEG monomer units, the amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M), and SG comprises N-acetylgalactosamine.

[0312] In some embodiments of a vaccine for inducing tolerance, amphiphile S comprises a second or third generation dendrimer, B comprises 4 to 36 PEG monomer units, amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M), and SG comprises N-acetylgalactosamine.

[0313] In some embodiments of the vaccine for inducing tolerance, the amphiphile S comprises a second or third generation dendrimer, the amphiphile H comprises a polymer of para-amino-phenylalanine, and the amphiphile SG comprises N-acetylgalactosamine.

[0314] In some embodiments of the vaccine for inducing tolerance, B comprises 4 to 36 PEG monomer units, the amphiphile H comprises a polymer of para-amino-phenylalanine, and SG comprises N-acetylgalactosamine.

[0315] In some embodiments of the vaccine for inducing tolerance, amphiphile S comprises a second or third generation dendrimer, B comprises 4 to 36 PEG monomer units, amphiphile H comprises a polymer of para-amino-phenylalanine, and SG comprises N-acetylgalactosamine.

[0316] In some embodiments of a vaccine for inducing tolerance, the amphiphile S comprises a second or third generation dendrimer, the amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and a reactive amino acid (N) comprising an imidazoquinoline, and the amphiphile SG comprises N-acetylgalactosamine.

[0317] In some embodiments of the vaccine for inducing tolerance, B comprises 4 to 36 PEG monomer units, the amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and a reactive amino acid (N) comprising an imidazoquinoline, and SG comprises N-acetylgalactosamine.

[0318] In some embodiments of a vaccine for inducing tolerance, amphiphile S comprises a second or third generation dendrimer, amphiphile B comprises 4 to 36 PEG monomer units, amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and a reactive amino acid (N) comprising an imidazoquinoline, and amphiphile SG comprises N-acetylgalactosamine.

[0319] In some embodiments of the vaccine for inducing tolerance, the amphiphile S comprises a second or third generation dendrimer, the amphiphile H comprises a poly(amino acid) of reactive amino acids (N) including tryptophan and imidazoquinoline, and the amphiphile SG comprises N-acetylgalactosamine.

[0320] In some embodiments of the vaccine for inducing tolerance, B comprises 4 to 36 PEG monomer units, the amphiphile H comprises a poly(amino acid) of reactive amino acids (N) including tryptophan and imidazoquinoline, and SG comprises N-acetylgalactosamine.

[0321] In some embodiments of the vaccine for inducing tolerance, amphiphile S comprises a second or third generation dendrimer, amphiphile B comprises 4 to 36 PEG monomer units, amphiphile H comprises a poly(amino acid) of reactive amino acids (N) including tryptophan and imidazoquinoline, and amphiphile SG comprises N-acetylgalactosamine.

[0322] In some embodiments of the vaccine for inducing tolerance, the amphiphile S comprises a second or third generation dendrimer, B comprises 4 to 36 PEG monomer units, and the peptide antigen conjugate comprises an enzyme-degradable linker.

[0323] In some embodiments of the vaccine for inducing tolerance, the amphiphile S comprises a second or third generation dendrimer, the amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M), and the peptide antigen conjugate comprises an enzymatically degradable linker.

[0324] In some embodiments of the vaccine for inducing tolerance, B comprises 4 to 36 PEG monomer units, the amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M), and the peptide-antigen conjugate comprises an enzymatically degradable linker.

[0325] In some embodiments of the vaccine for inducing tolerance, the amphiphile S comprises a second or third generation dendrimer, the amphiphile B comprises 4 to 36 PEG monomer units, the amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M), and the peptide-antigen conjugate comprises an enzymatically degradable linker.

[0326] In some embodiments of the vaccine for inducing tolerance, the amphiphile S comprises a second or third generation dendrimer, the amphiphile H comprises a polymer of para-amino-phenylalanine, and the peptide antigen conjugate comprises an enzyme-degradable linker.

[0327] In some embodiments of the vaccine for inducing tolerance, B comprises 4 to 36 PEG monomer units, the amphiphile H comprises a polymer of para-amino-phenylalanine, and the peptide-antigen conjugate comprises an enzyme-degradable linker.

[0328] In some embodiments of the vaccine for inducing tolerance, the amphiphile S comprises a second or third generation dendrimer, the amphiphile B comprises 4 to 36 PEG monomer units, the amphiphile H comprises a polymer of para-amino-phenylalanine, and the peptide-antigen conjugate comprises an enzyme-degradable linker.

[0329] In some embodiments of the vaccine for inducing tolerance, the amphiphile S comprises a second or third generation dendrimer, the amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and a reactive amino acid (N) comprising an imidazoquinoline, and the peptide-antigen conjugate comprises an enzyme-degradable linker.

[0330] In some embodiments of the vaccine for inducing tolerance, B comprises 4 to 36 PEG monomer units, the amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and a reactive amino acid (N) including an imidazoquinoline, and the peptide-antigen conjugate comprises an enzyme-degradable linker.

[0331] In some embodiments of the vaccine for inducing tolerance, the amphiphile S comprises a second or third generation dendrimer, the amphiphile B comprises 4 to 36 PEG monomer units, the amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and a reactive amino acid (N) including an imidazoquinoline, and the peptide-antigen conjugate comprises an enzyme-degradable linker.

[0332] In some embodiments of the vaccine for inducing tolerance, the amphiphile S comprises a second or third generation dendrimer, the amphiphile H comprises a poly(amino acid) of reactive amino acids (N) including tryptophan and imidazoquinoline, and the peptide-antigen conjugate comprises an enzyme-degradable linker.

[0333] In some embodiments of the vaccine for inducing tolerance, B comprises 4 to 36 PEG monomer units, the amphiphile H comprises a poly(amino acid) of reactive amino acids (N) including tryptophan and imidazoquinoline, and the peptide-antigen conjugate comprises an enzyme-degradable linker.

[0334] In some embodiments of the vaccine for inducing tolerance, the amphiphile S comprises a second or third generation dendrimer, the amphiphile B comprises 4 to 36 PEG monomer units, the amphiphile H comprises a poly(amino acid) of reactive amino acids (N) including tryptophan and imidazoquinoline, and the peptide-antigen conjugate comprises an enzyme-degradable linker.

[0335] In some embodiments of the vaccine for inducing tolerance, the amphiphile S 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) comprising a hydrophobic amino acid (M).

[0336] In some embodiments of a vaccine for inducing tolerance, the amphiphile S comprises a second or third generation dendrimer, the amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M), and the peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0337] 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).

[0338] In some embodiments of the vaccine for inducing tolerance, the amphiphile S comprises a second or third generation dendrimer, B comprises 4 to 36 PEG monomer units, the amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M), and the peptide-antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0339] In some embodiments of the vaccine for inducing tolerance, the amphiphile S comprises a second or third generation dendrimer, the amphiphile H comprises a polymer of para-amino-phenylalanine, and the peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0340] In some embodiments of the vaccine for inducing tolerance, B comprises 4-36 PEG monomer units, H of the amphiphile comprises a polymer of para-amino-phenylalanine, and H of the peptide-antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0341] In some embodiments of the vaccine for inducing tolerance, the amphiphile S comprises a second or third generation dendrimer, B comprises 4 to 36 PEG monomer units, the amphiphile H comprises a polymer of para-amino-phenylalanine, and the peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0342] In some embodiments of a vaccine for inducing tolerance, the amphiphile S comprises a second or third generation dendrimer, the amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and a reactive amino acid (N) comprising an imidazoquinoline, and the peptide-antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0343] 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) comprising an imidazoquinoline, and H of the peptide-antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0344] In some embodiments of the vaccine for inducing tolerance, the amphiphile S comprises a second or third generation dendrimer, B comprises 4 to 36 PEG monomer units, the amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and a reactive amino acid (N) comprising an imidazoquinoline, and the peptide-antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0345] In some embodiments of a vaccine for inducing tolerance, the amphiphile S comprises a second or third generation dendrimer, the amphiphile H comprises a poly(amino acid) of reactive amino acids (N) including tryptophan and imidazoquinoline, and the peptide antigen conjugate H comprises a poly(amino acid) including a hydrophobic amino acid (M).

[0346] In some embodiments of the vaccine for inducing tolerance, B comprises 4 to 36 PEG monomer units, the amphiphile H comprises a poly(amino acid) of reactive amino acids (N) including tryptophan and imidazoquinoline, and the peptide-antigen conjugate H comprises a poly(amino acid) including a hydrophobic amino acid (M).

[0347] In some embodiments of the vaccine for inducing tolerance, the amphiphile S comprises a second or third generation dendrimer, the amphiphile B comprises 4 to 36 PEG monomer units, the amphiphile H comprises a poly(amino acid) of reactive amino acids (N) including tryptophan and imidazoquinoline, and the peptide antigen conjugate H comprises a poly(amino acid) including a hydrophobic amino acid (M).

[0348] In some embodiments of the vaccine for inducing tolerance, the amphiphile S 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) comprising a hydrophobic amino acid (M).

[0349] In some embodiments of a vaccine for inducing tolerance, the amphiphile S comprises a second or third generation dendrimer, the amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M), the SG comprises N-acetylgalactosamine, and the peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0350] In some embodiments of the vaccine for inducing tolerance, B comprises 4-36 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).

[0351] In some embodiments of a vaccine for inducing tolerance, amphiphile S comprises a second or third generation dendrimer, B comprises 4 to 36 PEG monomer units, amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M), SG comprises N-acetylgalactosamine, and peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0352] In some embodiments of a vaccine for inducing tolerance, the amphiphile S comprises a second or third generation dendrimer, the amphiphile H comprises a polymer of para-amino-phenylalanine, the amphiphile SG comprises N-acetylgalactosamine, and the peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0353] In some embodiments of the vaccine for inducing tolerance, B comprises 4-36 PEG monomer units, the amphiphile H comprises a polymer of para-amino-phenylalanine, SG comprises N-acetylgalactosamine, and the peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0354] In some embodiments of the vaccine for inducing tolerance, the amphiphile S comprises a second or third generation dendrimer, B comprises 4 to 36 PEG monomer units, the amphiphile H comprises a polymer of para-amino-phenylalanine, SG comprises N-acetylgalactosamine, and the peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0355] In some embodiments of a vaccine for inducing tolerance, the amphiphile S comprises a second or third generation dendrimer, the amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and a reactive amino acid (N) comprising an imidazoquinoline, the amphiphile SG comprises N-acetylgalactosamine, and the peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0356] 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) comprising an imidazoquinoline, SG comprises N-acetylgalactosamine, and H of the peptide-antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0357] In some embodiments of a vaccine for inducing tolerance, amphiphile S comprises a second or third generation dendrimer, B comprises 4 to 36 PEG monomer units, amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and a reactive amino acid (N) comprising an imidazoquinoline, SG comprises N-acetylgalactosamine, and peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0358] In some embodiments of a vaccine for inducing tolerance, the amphiphile S comprises a second or third generation dendrimer, the amphiphile H comprises a poly(amino acid) of reactive amino acids (N) including tryptophan and imidazoquinoline, the SG comprises N-acetylgalactosamine, and the peptide antigen conjugate H comprises a poly(amino acid) including a hydrophobic amino acid (M).

[0359] In some embodiments of the vaccine for inducing tolerance, B comprises 4-36 PEG monomer units, the amphiphile H comprises a poly(amino acid) of reactive amino acids (N) including tryptophan and imidazoquinoline, SG comprises N-acetylgalactosamine, and the peptide antigen conjugate H comprises a poly(amino acid) including a hydrophobic amino acid (M).

[0360] In some embodiments of a vaccine for inducing tolerance, amphiphile S comprises a second or third generation dendrimer, B comprises 4 to 36 PEG monomer units, amphiphile H comprises a poly(amino acid) of reactive amino acids (N) including tryptophan and imidazoquinoline, SG comprises N-acetylgalactosamine, and peptide antigen conjugate H comprises a poly(amino acid) including a hydrophobic amino acid (M).

[0361] 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) comprising a hydrophobic amino acid (M).

[0362] In some embodiments of the vaccine for inducing tolerance, the amphiphile S comprises a second or third generation dendrimer, the amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M), and the peptide antigen conjugate comprises an enzymatically degradable linker, and the peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0363] 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 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).

[0364] In some embodiments of the vaccine for inducing tolerance, the amphiphile S 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) comprising a hydrophobic amino acid (M).

[0365] In some embodiments of a vaccine for inducing tolerance, the amphiphile S comprises a second or third generation dendrimer, the amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M), the amphiphile comprises amino-hexanoic acid, and the peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0366] 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).

[0367] In some embodiments of a vaccine for inducing tolerance, the amphiphile S comprises a second or third generation dendrimer, B comprises 4 to 36 PEG monomer units, the amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M), the amphiphile comprises amino-hexanoic acid, and the peptide-antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0368] In some embodiments of a vaccine for inducing tolerance, the amphiphile S comprises a second or third generation dendrimer, the amphiphile H comprises a polymer of para-amino-phenylalanine, the amphiphile comprises amino-hexanoic acid, and the peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0369] In some embodiments of the vaccine for inducing tolerance, B comprises 4 to 36 PEG monomer units, the amphiphile H comprises a polymer of para-amino-phenylalanine, the amphiphile comprises amino-hexanoic acid, and the peptide-antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0370] In some embodiments of a vaccine for inducing tolerance, the amphiphile S comprises a second or third generation dendrimer, B comprises 4 to 36 PEG monomer units, the amphiphile H comprises a polymer of para-amino-phenylalanine, the amphiphile comprises amino-hexanoic acid, and the peptide-antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0371] In some embodiments of a vaccine for inducing tolerance, the amphiphile S comprises a second or third generation dendrimer, the amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and a reactive amino acid (N) comprising an imidazoquinoline, the amphiphile comprises an amino-hexanoic acid, and the peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0372] 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) comprising an imidazoquinoline, the amphiphile comprises an amino-hexanoic acid, and H of the peptide-antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0373] In some embodiments of a vaccine for inducing tolerance, the amphiphile S comprises a second or third generation dendrimer, B comprises 4 to 36 PEG monomer units, the amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and a reactive amino acid (N) comprising an imidazoquinoline, the amphiphile comprises amino-hexanoic acid, and the peptide-antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0374] In some embodiments of a vaccine for inducing tolerance, the amphiphile S comprises a second or third generation dendrimer, the amphiphile H comprises a poly(amino acid) of reactive amino acids (N) including tryptophan and imidazoquinoline, the amphiphile comprises amino-hexanoic acid, and the peptide antigen conjugate H comprises a poly(amino acid) including a hydrophobic amino acid (M).

[0375] In some embodiments of the vaccine for inducing tolerance, B comprises 4-36 PEG monomer units, the amphiphile H comprises a poly(amino acid) of reactive amino acids (N) including tryptophan and imidazoquinoline, the amphiphile comprises amino-hexanoic acid, and the peptide-antigen conjugate H comprises a poly(amino acid) including a hydrophobic amino acid (M).

[0376] In some embodiments of a vaccine for inducing tolerance, the amphiphile S comprises a second or third generation dendrimer, B comprises 4 to 36 PEG monomer units, the amphiphile H comprises a poly(amino acid) of reactive amino acids (N) including tryptophan and imidazoquinoline, the amphiphile comprises amino-hexanoic acid, and the peptide antigen conjugate H comprises a poly(amino acid) including a hydrophobic amino acid (M).

[0377] In some embodiments of the vaccine for inducing tolerance, the amphiphile S comprises a second or third generation dendrimer, B comprises 4 to 36 PEG monomer units, 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).

[0378] In some embodiments of a vaccine for inducing tolerance, the amphiphile S comprises a second or third generation dendrimer, the amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M), the dendrimer monomer comprises a hydroxy acid and an amino alcohol, and the peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0379] In some embodiments of the vaccine for inducing tolerance, B comprises 4-36 PEG monomer units, the amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M), the dendrimer monomer comprises a hydroxy acid and an amino alcohol, and the peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0380] In some embodiments of a vaccine for inducing tolerance, the amphiphile S comprises a second or third generation dendrimer, B comprises 4 to 36 PEG monomer units, the amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M), the dendrimer monomer comprises a hydroxy acid and an amino alcohol, and the peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0381] In some embodiments of a vaccine for inducing tolerance, the amphiphile S comprises a second or third generation dendrimer, the amphiphile H comprises a polymer of para-amino-phenylalanine, the dendrimer monomer comprises a hydroxy acid and an amino alcohol, and the peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0382] In some embodiments of the vaccine for inducing tolerance, B comprises 4-36 PEG monomer units, the amphiphile H comprises a polymer of para-amino-phenylalanine, the dendrimer monomer comprises a hydroxy acid and an amino alcohol, and the peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0383] In some embodiments of a vaccine for inducing tolerance, the amphiphile S comprises a second or third generation dendrimer, B comprises 4 to 36 PEG monomer units, the amphiphile H comprises a polymer of para-amino-phenylalanine, the dendrimer monomer comprises a hydroxy acid and an amino alcohol, and the peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0384] In some embodiments of a vaccine for inducing tolerance, the amphiphile S comprises a second or third generation dendrimer, the amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and a reactive amino acid (N) comprising an imidazoquinoline, the dendrimer monomer comprises a hydroxy acid and an amino alcohol, and the peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0385] In some embodiments of the vaccine for inducing tolerance, B comprises 4-36 PEG monomer units, the amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and a reactive amino acid (N) including an imidazoquinoline, the dendrimer monomer comprises a hydroxy acid and an amino alcohol, and the peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0386] In some embodiments of a vaccine for inducing tolerance, the amphiphile S comprises a second or third generation dendrimer, B comprises 4 to 36 PEG monomer units, the amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and a reactive amino acid (N) including an imidazoquinoline, the dendrimer monomer comprises a hydroxy acid and an amino alcohol, and the peptide-antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0387] In some embodiments of a vaccine for inducing tolerance, the amphiphile S comprises a second or third generation dendrimer, the amphiphile H comprises a poly(amino acid) of reactive amino acids (N) including tryptophan and imidazoquinoline, the dendrimer monomer comprises a hydroxy acid and an amino alcohol, and the peptide antigen conjugate H comprises a poly(amino acid) including a hydrophobic amino acid (M).

[0388] In some embodiments of the vaccine for inducing tolerance, B comprises 4-36 PEG monomer units, the amphiphile H comprises a poly(amino acid) of reactive amino acids (N) including tryptophan and imidazoquinoline, the dendrimer monomer comprises a hydroxy acid and an amino alcohol, and the peptide antigen conjugate H comprises a poly(amino acid) including a hydrophobic amino acid (M).

[0389] In some embodiments of a vaccine for inducing tolerance, the amphiphile S comprises a second or third generation dendrimer, B comprises 4 to 36 PEG monomer units, the amphiphile H comprises a poly(amino acid) of reactive amino acids (N) including tryptophan and imidazoquinoline, the dendrimer monomer comprises a hydroxy acid and an amino alcohol, and the peptide antigen conjugate H comprises a poly(amino acid) including a hydrophobic amino acid (M).

[0390] The present disclosure also relates to 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, During the ceremony, A is a peptide antigen, E1 is the N-terminal extension, E2 is a C-terminal extension, H, independently at each occurrence, is a hydrophobic block; and one or more drug molecules (D) are optionally attached to each H, either directly or via a suitable linker, X1; S is a solubilizing block, B is a spacer, U is, independently at each occurrence, a linker; [ ] means that the group is optional; - denotes that two adjacent groups are directly linked to each other by a covalent bond or indirectly linked to each other via a suitable linker X; wherein the amphiphile comprises a dendron amplifier, and at least one A comprises a sequence in which one or more cysteine ​​residues are replaced with alpha-aminobutyric acid and / or one or more methionine residues are replaced with norleucine.

[0391] In some embodiments of the vaccine, the amphiphile S comprises a carboxylic acid, hi other embodiments of the vaccine, the amphiphile S comprises succinic acid or beta-alanine.

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

[0393] In some embodiments of the vaccine, the average net charge of at least one peptide-antigen conjugate is positive at physiological pH, and the molar ratio of peptide-antigen conjugate to amphiphile is about 4:1 to about 2:1, or about 1:2 to about 1:16, or about 1:2 to about 1:4, preferably about 1:1.

[0394] The present disclosure also relates to vaccines comprising at least one peptide antigen (A), wherein the at least one peptide antigen (A) comprises a sequence in which one or more cysteine ​​residues are replaced with alpha-aminobutyric acid and / or one or more methionine residues are replaced with norleucine.

[0395] In some embodiments of the vaccine, the vaccine further comprises a particulate delivery system selected from a lipid emulsion, a liposome, a PLGA particle, an inorganic salt particle, and a metal nanoparticle. In other embodiments of the vaccine, the vaccine further comprises at least one drug molecule (D) selected from an immunostimulant and a Treg-promoting immunomodulator.

[0396] The present disclosure also relates to 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, During the ceremony, H, independently at each occurrence, is a hydrophobic block; one or more drug molecules (D) are optionally attached to each H directly or via a suitable linker X1; A, independently at each occurrence, is a peptide antigen; E1 is, independently at each occurrence, an N-terminal extension, E2 is, independently at each occurrence, a C-terminal extension, U is, independently at each occurrence, a linker; where: (i) at least one A comprises alpha amino butyric acid and / or norleucine; (ii) at least one A is selected from a tumor antigen, at least one D is present and is selected from an agonist of TLR-7 / 8, and the vaccine further comprises a second drug molecule (D2) selected from an inhibitor of mTOR; (iii) at least one A is a glycopeptide; or (iv) at least one A is selected from autoantigens, allergens, and alloantigens; at least one D is present and is selected from ATP-competitive mTOR inhibitors; and [ ] indicates that the group is optional; - indicates that two adjacent groups are directly linked to each other by a covalent bond, or indirectly linked to each other via a suitable linker X.

[0397] In some embodiments of the vaccine, at least one peptide antigen conjugate comprises at least one A selected from a tumor antigen.

[0398] In some embodiments of the vaccine, at least one D is selected from an agonist of TLR-3, TLR-7, TLR-8, TLR-9, and STING.

[0399] In some embodiments of the vaccine, each H of the amphiphile and / or peptide antigen conjugate comprises a poly(amino acid) comprising a monomer of a reactive amino acid (N), wherein the monomer comprises D selected from an agonist of TLR-7 / 8.

[0400] In some embodiments of the vaccine, D2 is present and is selected from rapamycin, tacrolimus, everolimus, RAD001, CCI-779, and AP23573.

[0401] In some embodiments of the vaccine, the molar ratio of peptide antigen conjugate to D2 is about 20:1 to 1:2, or about 10:1 to about 1:1, or about 4:1 to about 2:1, preferably about 1:1.

[0402] In some embodiments of the vaccine, at least one A is a glycopeptide, e.g., A is a glycopeptide 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), wherein: * is an O-linked glycan, and each occurrence is independently selected from sialyl Lewis x, sialyl Lewis a, Lewis y, Lewis x, Tn, sTn, TF, and sTF.

[0403] In some embodiments of the vaccine, where A is a glycopeptide, S is absent. In other embodiments, S is present.

[0404] In some embodiments of the vaccine, the vaccine further comprises an amphiphile having the formula S-[B]-[U]-H, where S is a solubilizing block; B is a spacer, H is a hydrophobic block, U is a linker, [ ] means that the group is optional; - denotes that two adjacent groups are directly linked to each other by a covalent bond or indirectly linked to each other via a suitable linker X; Here, the amphiphilic material S comprises a dendron amplifier.

[0405] In some embodiments of the vaccine, the amphiphile, S, comprises two or more solubilizing groups (SG) independently selected from an amine, a carboxylic acid, or a sugar molecule, wherein the sugar molecule is 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.

[0406] In some embodiments of the vaccine, at least one peptide antigen conjugate comprises at least one A selected from an autoantigen, an alloantigen, and an allergen.

[0407] In some embodiments of the vaccine, the vaccine comprises an inhibitor of mTOR, RORgt, CDK8 / 19, and HDAC, as well as an inhibitor of AHR, RAR, and A 2a and further comprising at least one D selected from the group consisting of agonists of

[0408] In some embodiments of the vaccine, the vaccine comprises an inhibitor of mTOR, RORgt, CDK8 / 19, and HDAC, AHR, RAR, and A 2a and an immunostimulant selected from agonists of NLR, CLR, TLR and STING, provided that D and D2 bind to different receptors.

[0409] In some embodiments of the vaccine, D2 is selected from an agonist of an NLR, a CLR, a TLR, and a STING. In other embodiments, D2 is selected from an agonist of TLR-3, TLR-7, TLR-8, TLR-7 / 8, TLR-9, and STING. In other embodiments, D2 is selected from RNA and an imidazoquinoline agonist of TLR-7, TLR-8, and TLR-7 / 8.

[0410] In some embodiments of the vaccine, the vaccine comprises an inhibitor of mTOR, RORgt, CDK8 / 19, and HDAC, AHR, RAR, and A 2a and an immunostimulant selected from agonists of NLR, CLR, TLR and STING, provided that D, D2 and D3 bind to different receptors.

[0411] In some embodiments of the vaccine, 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.

[0412] In some embodiments of the vaccine, the molar ratio of total peptide antigen conjugate to at least one D is about 20:1 to 1:2, or about 10:1 to about 1:1, or about 4:1 to about 2:1, preferably about 1:1.

[0413] In a preferred embodiment of the vaccine for inducing tolerance, the vaccine comprises at least one peptide-antigen conjugate having a formula selected from PEG-[E1]-A-[E2]-[U]-H and H-[U]-[E1]-A-[E2]-PEG, During the ceremony, H, independently at each occurrence, is a hydrophobic block; one or more drug molecules (D) are optionally attached to each H directly or via a suitable linker X1; A, independently at each occurrence, is a peptide antigen; E1 is, independently at each occurrence, an N-terminal extension, E2 is, independently at each occurrence, a C-terminal extension, U is, independently at each occurrence, a linker; wherein at least one A is selected from autoantigens, allergens and alloantigens; at least one D is present and is selected from ATP-competitive mTOR inhibitors; [ ] denotes that the group is optional; and - denotes that two adjacent groups are directly covalently bonded to each other or indirectly bonded to each other via a suitable linker, X.

[0414] In some embodiments of the vaccine, 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.

[0415] In some embodiments of the vaccine, the vaccine further comprises an amphiphile having the formula S-[B]-[U]-H, where S is a solubilizing block; B is a spacer, H is a hydrophobic block, U is a linker, [ ] means that the group is optional; - denotes that two adjacent groups are directly linked to each other by a covalent bond or indirectly linked to each other via a suitable linker X; Here, the amphiphilic material S comprises a dendron amplifier.

[0416] In some embodiments of the vaccine, the amphiphile S comprises two or more solubilizing groups (SG) independently selected from a carboxylic acid, a phosphoserine, and a sugar molecule, wherein the sugar molecule is independently selected from mannose, glucose, glucosamine, N-acetylglucose, galactose, galactosamine, N-acetylgalactosamine, and an agonist of CD22a.

[0417] In some embodiments of the vaccine, the peptide-antigen conjugate has a net positive charge of about +1 to about +10 at physiological pH, while in other embodiments, the peptide-antigen conjugate has a net positive charge of about +2 to about +6 or about +3 to about +5 at physiological pH.

[0418] In some embodiments of the vaccine, an amphiphile is present and the molar ratio of peptide antigen conjugate to amphiphile is about 4:1 to 1:20, preferably about 1:1.

[0419] In some embodiments of the vaccine, the amphiphile comprises a carboxylic acid and has a net negative charge, hi other embodiments, the amphiphile comprises a carboxylic acid selected from beta-alanine and succinic acid.

[0420] In some embodiments of the vaccine, the average net charge of at least one peptide-antigen conjugate is positive at physiological pH, and the molar ratio of peptide-antigen conjugate to amphiphile is about 4:1 to about 2:1, or about 1:2 to about 1:16, or about 1:2 to about 1:4. In certain preferred embodiments, the molar ratio is about 1:1.

[0421] In some embodiments of the vaccine, the vaccine comprises particles further comprising an amphiphile and one or more peptide antigen conjugates. In a preferred embodiment of the vaccine, the vaccine comprises particles 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 the peptide antigen, S is a solubilizing block, E1 and E2 are N-terminal and C-terminal extensions, respectively, B is a spacer, U is a linker molecule, H is a hydrophobic block, [ ] denotes that the group is optional, - denotes that two adjacent groups are directly linked to each other by a covalent bond or indirectly linked to each other via a suitable linker X, and U and H of the amphiphile and the peptide antigen conjugate may be the same or different or may comprise one or more of the same functional groups or moieties.

[0422] In some embodiments of the vaccine, the amphiphile and / or peptide antigen conjugate further comprises one or more drug molecules (D), which may be either 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) may be incorporated 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) may be in the form of a drug molecule conjugate (i.e., D-[U]-H or HD) incorporated with the amphiphile and / or peptide antigen conjugate (e.g., D-H + ​​S-[B]-[U]-H + PEG-[E1]-A-[E2]-[U]-H). Preferred vaccine compositions further comprising a drug molecule (D) are described throughout this specification. D is attached directly or indirectly to an adjacent group as a side chain or as part of a side chain group.

[0423] In a preferred embodiment of the vaccine, the vaccine comprises particles comprising an amphiphile and one or more peptide antigen conjugates, further comprising a drug molecule (D) selected from an immunomodulatory agent. The drug molecule (D) selected from an immunomodulatory agent may be linked directly to the hydrophobic block (H) of the amphiphile and / or peptide antigen conjugate or indirectly via X1 (e.g., S-[B]-[U]-HD and / or PEG-[E1]-A-[E2]-[U]-HD), or the drug molecule (D) may be admixed 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) may be in the form of a drug molecule conjugate (i.e., D-[U]-H or HD) admixed with the amphiphile and peptide antigen conjugate (e.g., D-H + ​​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.

[0424] In some embodiments of vaccines for treating or preventing autoimmune diseases, the peptide antigen conjugate comprises an antigen (A) selected from a self-antigen (sometimes referred to as an autoantigen). In some embodiments of vaccines for treating or preventing allergies, the peptide antigen conjugate comprises an antigen (A) selected from an allergen. In some embodiments of vaccines for treating or preventing transplant rejection, the peptide antigen conjugate comprises an antigen (A) selected from an alloantigen. In some embodiments of vaccines for treating or preventing cancer, the peptide antigen conjugate comprises an antigen (A) selected from a self-antigen, a neoantigen, or a viral antigen. In some embodiments of vaccines for treating infectious diseases, the peptide antigen conjugate comprises an antigen (A) selected from a virus, bacterium, protozoan, or fungus. Preferred antigens for treating different diseases as well as preferred methods for selecting antigens are described throughout this specification.

[0425] It has been discovered that particles comprising certain compositions of amphiphiles have particular utility in 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 diseases.

[0426] 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 having the formula S-[B]-[U]-H and a drug, D, where S is a solubilizing block, B is a spacer, U is a linker molecule, H is a hydrophobic block, [ ] denotes that the group is optional, and the drug, D, is associated with the particle by covalent or non-covalent interactions.

[0427] In some embodiments, drug molecules (D), if present, are linked to the hydrophobic block (H) of the amphiphile (e.g., S-[B]-[U]-H-D), where one or more Ds are attached directly or indirectly through X1 to adjacent groups at the termini(s) or as part of a side group. In other embodiments, drug molecules are intermixed with the amphiphile (e.g., D + S-[B]-[U]-H) or linked to the hydrophobic block (H) and intermixed with the amphiphile (e.g., D-[B]-[U]-H + S-[B]-[U]-H, or H-D + S-[B]-[U]-H), and the drug is incorporated into particles formed by the amphiphile. Preferred compositions of cancer therapeutics comprising an amphiphile and at least one chemotherapeutic agent and / or immunostimulant are described throughout this specification.

[0428] 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.

[0429] 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, During the ceremony, H is a hydrophobic block, one or more drug molecules (D) are optionally attached to each H directly or via a suitable linker X1; A is a peptide antigen, E1 is the N-terminal extension, E2 is a C-terminal extension, U is a linker, U1 is a linker precursor, [ ] means that the group is optional; - indicates that two adjacent groups are directly linked to each other by a covalent bond, or indirectly linked to each other via a suitable linker X.

[0430] In some embodiments, the peptidic antigen fragment has the formula A-[E2]-PEG.

[0431] In some embodiments, E1 and / or E2 are present and are selected from cathepsin-cleavable tetrapeptides of the formula P4-P3-P2-P1.

[0432] 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.

[0433] In some embodiments, the peptide antigen (A) comprises at least one amino acid selected from norleucine and alpha-aminobutyric acid.

[0434] Disclosed herein in some embodiments is a vaccine comprising 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; During the ceremony, H is a hydrophobic block, one or more drug molecules (D) are optionally attached to each H directly or via a suitable linker X1; A is a peptide antigen, E1 is the N-terminal extension, E2 is a C-terminal extension, U is a linker, U1 is a linker precursor, [ ] means that the group is optional; - indicates that two adjacent groups are directly linked to each other by a covalent bond, or indirectly linked to each other via a suitable linker X.

[0435] In some embodiments, the vaccine for inducing tolerance can be used to prevent or treat autoimmune diseases, including, but not limited to, multiple sclerosis, anti-MOG, celiac disease (including intractable disease), type 1 diabetes, vitiligo, autoimmune hepatitis, neuromyelitis optica, autoimmune uveitis, rheumatoid arthritis, myasthenia gravis, Lambert-Eaton syndrome, Graves' disease, optic neuropathy, immune thrombocytopenic purpura, pemphigus vulgaris, bullous pemphigoid, Gutpasture's syndrome, eczema, Sjögren's syndrome, achalasia, and myeloma. These include, but are not limited to, inflammatory bowel disease including psoriasis, dermatomyositis, systemic sclerosis, psoriasis, Crohn's disease and ulcerative colitis, primary sclerosing cholangitis, Takayasu's arteritis, giant cell arteritis, polyarteritis nodosa, Kawasaki disease, anti-GBM disease, various vasculitides including, but not limited to, ANCA vasculitis, systemic lupus erythematosus, amyotrophic lateral sclerosis (ALS), Behcet's disease, and birch pollen allergy.

[0436] 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 PEG-[E1]-A-[E2]-[U]-H and H-[U]-[E1]-A-[E2]-PEG, and an amphiphile having the formula S-[B]-[U]-H; During the ceremony, A is a peptide antigen, E1 is the N-terminal extension, E2 is a C-terminal extension, H, independently at each occurrence, is a hydrophobic block; and one or more drug molecules (D) are optionally attached to each H, either directly or via a suitable linker, X1; S is a solubilizing block, B is a spacer, U is, independently at each occurrence, a linker; [ ] means that the group is optional; - denotes that two adjacent groups are directly linked to each other by a covalent bond or indirectly linked to each other via a suitable linker X; wherein the amphiphile comprises a dendron amplifier and the at least one peptide antigen is selected from an autoantigen or a tumor antigen.

[0437] In one embodiment of the method of treating an autoimmune disease, the vaccine is administered intravenously, subcutaneously, or intramuscularly.

[0438] The present disclosure also relates to a method for improving the efficacy and / or tolerability of a vaccine, the method comprising administering to a subject 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; During the ceremony, A is a peptide antigen, E1 is the N-terminal extension, E2 is a C-terminal extension, H, independently at each occurrence, is a hydrophobic block; and one or more drug molecules (D) are optionally attached to each H, either directly or via a suitable linker, X1; S is a solubilizing block, B is a spacer, U is, independently at each occurrence, a linker; [ ] means that the group is optional; - indicates that two adjacent groups are directly bonded to each other by a covalent bond or indirectly bonded to each other via a suitable linker; wherein the amphiphile comprises a dendron amplifier and the at least one peptide antigen is selected from an autoantigen.

[0439] 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 in a subject is directed against an antigen, which can be an autoantigen, an alloantigen, a foreign antigen (e.g., an allergen), a drug molecule, or a device.

[0440] The present disclosure also relates to a method of inducing an immune response in a subject in need thereof, the method 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.

[0441] In some embodiments, the T cell response in the subject is increased compared to administration of only at least one dose of the first vaccine (V1).

[0442] In some embodiments, the T cell response in the subject is increased compared to administration of only at least one dose of the second vaccine (V2).

[0443] In some embodiments, one dose of V1 is administered at time 1 (V1T1). In other embodiments, two doses of V1 are administered at time 1 (V1T1) and time 2 (V1T2). In other embodiments, three doses of V1 are administered at time 1 (V1T1), time 2 (V1T2), and time 3 (V1T3).

[0444] In some embodiments, one dose of V2 is administered at time 1 (V2T1). In other embodiments, two doses of V2 are administered at time 1 (V2T1) and time 2 (V2T2). In other embodiments, three doses of V2 are administered at time 1 (V2T1), time 2 (V2T2), and time 3 (V2T3).

[0445] In some embodiments, V1 is administered by intramuscular or intravenous route.

[0446] In some embodiments, V2 is administered by the intravenous route.

[0447] In some embodiments, the first dose of V2 is administered 1 to 6 weeks after the last dose of V1. In other embodiments, the first dose of V2 is administered 1 to 12 weeks after the last dose of V1.

[0448] In some embodiments, V2 is an adenovirus vector vaccine.

[0449] In some embodiments, the adenovirus encodes a peptide antigen (A) of V1.

[0450] In some embodiments, V2 is a ChAdOx vaccine.

[0451] Linker The term linker refers to any molecule that joins together any two or more molecules (or "moieties"), such as any two or more components of an amphiphile, a peptide-antigen conjugate, or a drug conjugate, and may further perform any one or more of the following functions: I) increasing or decreasing aqueous solubility, II) increasing the distance between any two components, III) imparting rigidity or flexibility, or IV) modulating the degradation rate of the linkage between any two or more different molecules. As used herein, the term "linker" may 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).

[0452] Linkers with particular utility are listed, and specific preferred compositions of these listed 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. They may be included between a 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 resulting from the reaction of linker precursor 1 ("U1"), linked either directly or indirectly to the solubilizing block or drug (D) via spacer (B), with linker precursor 2 ("U2") on the hydrophobic block (H) is referred to as linker U. Suitable linker X refers to any linker suitable for linking two or more adjacent groups. A preferred linker for connecting a drug molecule (D) to a hydrophobic block (H) is designated X1. A preferred linker for connecting an aryl or heteroaryl group to a hydrophobic block is designated X2. A preferred linker used to connect a reactive functional group ("FG4") to the pharmacophore of a drug molecule (D) is designated X3. A preferred linker for connecting a charged group to a hydrophobic block (H) is designated X4. A preferred linker for connecting SG to S is designated X5.

[0453] Linker may use covalent or non-covalent means to connect any two or more components.In a preferred embodiment, linker may connect, i.e., link, any two components via covalent bond.Covalent bond is the preferred bond used to connect any two components, and ensures that no component can be readily dispersed from other components after administration to a subject.

[0454] There are many suitable linkers known to those skilled in the art, including, but not limited to, straight or branched chain carbon linkers, heterocyclic carbon linkers, rigid aromatic linkers, flexible ethylene oxide linkers, peptide linkers, or combinations thereof, which, in the case of covalent linkers, further comprise two or more functional groups, which may be the same or different, used to link any two molecules, e.g., any two components of an amphiphile, a peptide antigen conjugate, and / or a drug conjugate, via a covalent bond.

[0455] In some embodiments, the carbon linker can include a lower alkyl linker, such as a C1-C18 alkane linker, e.g., a C1-C6 (i.e., 1-6 methylene units), which serves to increase the spacing between two or more molecules, i.e., different components, while longer chain alkane linkers can be used to impart hydrophobic properties. Alternatively, a hydrophilic linker, such as an ethylene oxide linker, may be used in place of the alkane linker to increase the spacing between any two or more heterogeneous molecules and increase aqueous solubility. In other embodiments, the linker can be a cyclic and / or aromatic compound, or a poly(aromatic) compound to impart rigidity. The linker molecule can include a hydrophilic or hydrophobic linker. In some embodiments, the linker includes a degradable peptide sequence that is cleavable by intracellular enzymes (e.g., cathepsins or immunoproteasomes).

[0456] When linking two components of amphiphiles, peptide-antigen conjugates, and drug conjugates, at least one of which comprises a peptide, it has been found that linkers containing two to seven methylene groups improve the binding of the two components. In a non-limiting example, increasing the number of methylene units between the amide and amine of the N-terminal amino acid of the peptide-based hydrophobic block (H) results in improved binding to other molecules, including U2, antigens (A), extensions E2, spacers (B), and solubilizing blocks (S) such as PEG and charged blocks (C). Thus, in preferred embodiments, the N-terminal amino acid of the poly(amino acid)-based hydrophobic block (H) contains two or more methylene units, typically two to seven, e.g., 1, 2, 3, 4, 5, 6, or 7 methylene units. For clarity, an amino acid with two methylene units is beta-alanine, and an amino acid with five 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.

[0457] In some embodiments, the linker may comprise poly(ethylene oxide) (PEG). The length of the linker depends on the purpose of the linker. For example, the length of a linker, such as a PEG linker, can be increased to separate two or more components, e.g., to reduce steric hindrance, or, in the case of a hydrophilic PEG linker, to improve aqueous solubility. Linkers, such as PEG, may be from about 1 to about 24 monomers in length, e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or more monomers in length. When used as a spacer (B), PEG can be up to 45 or more monomers in length, but typically 4 to 36 monomers in length.

[0458] In some embodiments, when the linker comprises a carbon chain, the linker may comprise a chain of from about 1 or 2 to about 18 carbons, e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18 carbons or more in length. In some embodiments, when the linker comprises a carbon chain, the linker may comprise a chain of from about 12 to about 20 carbons. In some embodiments, when the linker comprises a carbon chain, the linker may comprise a chain of 18 carbons or less, typically about 1 to 6 carbon atoms.

[0459] 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, may comprise any suitable functional group, including, but not limited to, amide, ester, ether, thioether, silyl ether, disulfide, carbamate, carbamide, hydrazide, hydrazone, acetal, and triazole.

[0460] In a non-limiting example of a covalent bond, a click chemistry reaction can result in a triazole that links, i.e., joins together, any two components of an amphiphile, a peptide-antigen conjugate, or a drug molecule conjugate. In some embodiments, the click chemistry reaction is a strain-promoted [3+2] azide-alkyne cycloaddition reaction. An alkyne group and an azide group may be provided on each molecule to be linked by "click chemistry." In some embodiments, an antigen (A) bearing an azide functional group is coupled to a hydrophobic block (H) with a suitable reactive group, such as an alkyne, e.g., dibenzylcyclooctyne (DBCO).

[0461] In some embodiments, an amine is provided on one molecule and may be linked to another molecule by reacting the amine with any suitable electrophilic group, such as a carboxylic acid, an acid chloride, an activated ester (e.g., an NHS ester), thereby resulting in an amide bond; the amine may be reacted with an alkene (via a Michael addition); the amine may be reacted with an aldehyde and a ketone (via a Schiff base); or the amine may be reacted with an activated carbonate or carbamate to give a carbamate.

[0462] In some embodiments, the linker is cleavable under intracellular conditions such that cleavage of the linker results in release of any component linked to the linker, for example, a drug molecule (D).

[0463] For example, the linker can be cleavable by an enzyme localized in an intracellular vesicle (e.g., in a lysosome or endosome or caveolae) or by an enzyme in the cytosol, such as a proteasome or immunoproteasome. The linker can be a peptide linker that is cleaved by a protease enzyme, including, but not limited to, a protease localized in an intracellular vesicle, such as, for example, a cathepsin in the lysosomal or endosomal compartment of a cell.

[0464] Peptide linkers are typically 1 to 10 amino acids long, e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more (e.g., up to 20) amino acids long, e.g., 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more. When used as a spacer (B), the peptide linker can be up to about 45 amino acids long. Certain dipeptides are known to be hydrolyzed by proteases, including cathepsins such as cathepsin B and D and plasmin (see, e.g., Dubowchik and Walker, 1999, Pharm. Therapeutics 83:67-123). For example, a peptide linker cleavable by the thiol-dependent protease cathepsin B can be used (e.g., Phe-Leu or Gly-Phe-Leu-Gly (SEQ ID NO: 1) linker). Other examples of such linkers are described, for example, in U.S. Patent No. 6,214,345, which is 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 using a Val-Cit linker). Note: For example amino acids and peptide examples provided throughout this specification (whether in the text of figures), it is understood that the peptides and amino acids are L-amino acids unless otherwise specified.

[0465] The cleavable peptide linker can be selected to facilitate 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 facilitate processing by intracellular proteases, such as cathepsins in intracellular vesicles or proteasomes or immunoproteasomes in the cytosolic space.

[0466] In some embodiments, to facilitate recognition by cathepsins, linkers comprising peptide sequences of the formula Pn...P4-P3-P2-P1 are used, where 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 linked to another molecule by an amide bond 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 (e.g., where the amide bond between P1 and P1' is hydrolyzed), which is C-terminal to the P1 residue. 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).

[0467] In some embodiments, linkers comprising peptide sequences are selected to promote recognition by the proteasome or immunoproteasome. Peptide sequences of the formula Pn...P4-P3-P2-P1 are selected to promote recognition by the proteasome or immunoproteasome, where P1 is selected from basic residues and hydrophobic branched residues, e.g., arginine, lysine, leucine, isoleucine, and valine, and 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 by 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 may be used alone or in combination with cathepsin-cleavable linkers. In some embodiments, the amino acids that promote immunoproteasomal processing are linked to a linker that promotes processing by endosomal proteases. Some suitable sequences for promoting immunoproteasomal cleavage are described in detail 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).

[0468] In certain preferred embodiments, the drug molecule (D) is linked to the hydrophobic block (H) via a linker X1 that comprises an enzymatically degradable peptide. Non-limiting examples are provided here: [ka] where D is a drug molecule, "linker" is any suitable linker molecule, j represents any integer, but j is typically 1 to 6 amino acids, e.g., 1, 2, 3, 4, 5, or 6 amino acids, and R 8is any suitable amino acid side chain, and the N-terminal amine of the peptide is linked, for example, either directly or via a U, to the N-terminus or C-terminus of the hydrophobic block (H) comprising a poly(amino acid), either directly, or via a U, or via a reactive monomer that makes up the hydrophobic block (H), and the square brackets "[ ]" indicate that the group is optional.

[0469] In certain preferred embodiments of a drug molecule linked to a hydrophobic block (H) via a linker X1 comprising an enzymatically degradable peptide, the drug molecule (D) is linked directly to the peptide by an amide bond as shown here: [ka]

[0470] In a non-limiting example of the above structure, when an N-terminal linker group is present and selected from beta-alanine, the structure is: [ka]

[0471] In some embodiments, the drug molecule (D) is linked to the peptide via a self-immolative carbamate linker. Non-limiting examples are provided here: [ka]

[0472] In the above example, if j is 4 and the amino acids are serine-lysine (Ac)-valine-norleucine, the structure is: [ka]

[0473] In some embodiments, the drug molecule (D) is linked to the hydrophobic block (H) via a sulfatase-degradable linker X1, where hydrolysis of the sulfate by sulfatase results in release of the drug molecule from the linker. Several arylsulfatase- and alkylsulfatase-degradable linkers have recently been described (see, e.g., Bargh, et al., 2020, Chem. Sci. 11, 2375). In some embodiments of the present disclosure, the drug molecule is linked to the hydrophobic block (H) via a sulfatase-degradable linker. Non-limiting examples are provided here for clarity: [ka] In the formula, D is a drug molecule, the "linker" is any suitable linker molecule linked, for example, directly or via a U or via a reactive monomer that constitutes the hydrophobic block (H), to the N-terminus or C-terminus of the hydrophobic block (H) comprising a poly(amino acid), and the square brackets "[ ]" indicate that the group is optional.

[0474] A non-limiting example of the above structure is shown here for clarity, where the "linker" is present and is selected from a short alkyl linker connected to the hydrophobic block via an amide. [ka]

[0475] In other embodiments, any two or more components may be linked together via a pH-sensitive linker X that is susceptible to hydrolysis under acidic conditions. Several pH-sensitive linkers are well known to those skilled in the art, including hydrazones, carbohydrazones, semicarbazones, thiosemicarbazones, cis-aconitic amides, 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).

[0476] In certain embodiments, 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., about pH 7.4, but undergoes more rapid hydrolysis at endosomal / lysosomal pH, e.g., about pH 5-6.5. In certain preferred embodiments, a drug molecule (D) is linked to the hydrophobic block (H) via a reactive monomer via a pH-sensitive bond, such as a hydrazone bond resulting from the reaction between a ketone and a hydrazine. A carbonyl-linked functional group, hydrazine, is sometimes referred to as a hydrazide, although hydrazine is intended to refer broadly to the -NH-NH group, including when linked to a carbonyl (e.g., C(O)-NH-NH). pH-sensitive bonds, such as hydrazones, offer the advantage that the bond is stable at physiological pH, e.g., about pH 7.4, but is hydrolyzed at lower pH values, such as the pH of intracellular vesicles.

[0477] In certain preferred embodiments, the drug molecules are linked by a ketone-containing linker X1 and have the formula: [ka] where D is any drug molecule, "linker" is any suitable linker molecule, y1 represents an integer from 1 to 6, preferably 4, and square brackets "[ ]" indicate that the group is optional, and the ketone in the above example is used to link the linker-linked drug molecule (D) to the reactive monomer via a hydrazone bond.

[0478] In the above example, if y1 is 4 and the drug molecules are directly linked via an amide bond (ie, there is no "linker"), the structure is: [ka]

[0479] In a preferred embodiment, the ketone-linked drug molecule is linked to the hydrophobic block (H) via a hydrazone or carbohydrazone bond. Non-limiting examples of drug molecules linked to glutamic acid-based reactive monomers (N) via hydrazone and carbohydrazone bonds are shown here: [ka]

[0480] In some embodiments, the drug molecule comprises a ketone and may be directly linked to the reactive monomer via a hydrazone or carbohydrazone.

[0481] In other embodiments, the linker comprises a bond that can be cleaved under reducing conditions, such as a reducible disulfide bond. Many different linkers used to introduce disulfide bonds are known in the art (see, e.g., Thorpe et al., 1987, Cancer Res. 47:5924-5931; Wawrzynczak et al., In Immunoconjugates: Antibody Conjugates in Radioimagery and Therapy of Cancer (C.W. Vogel ed., Oxford U. Press, 1987); Phillips et al., Cancer Res. 68:9280-9290, 2008). See also U.S. Patent No. 4,880,935.

[0482] In a preferred embodiment, the linker X1 connecting the hydrophobic block (H) and one or more drug molecules (D) is a short alkyl or PEG linker. In another preferred embodiment, the linker X1 connecting the hydrophobic block (H) and one or more drug molecules (D) is an enzyme-degradable linker, such as a cathepsin-degradable peptide or a sulfatase-degradable linker. In another preferred embodiment, the linker X1 connecting the hydrophobic block (H) and one or more drug molecules (D) comprises an enzyme-degradable peptide or a self-immolative linker.

[0483] X can be any suitable linker, but in preferred embodiments, the linker X connecting any two or more groups is a short alkyl (i.e., lower alkyl) or PEG linker, for example, a PEG linker having from about 1 to about 24 monomer units.

[0484] Extensions (E1 and E2) The optional N- and C-terminal extensions (El and E2) represent moieties linked to the N- and C-termini of the peptidic antigen (A), respectively. The N- and C-terminal extensions El and E2 may comprise any one or more of the following: amino acids, including unnatural amino acids, hydrophilic ethylene oxide monomers (e.g., PEG), hydrophobic alkane chains, etc., or combinations thereof. The N- and C-terminal extensions El and E2 are linked to the peptidic antigen (A) by any suitable means, for example, by an amide bond.

[0485] In some embodiments, the extensions (E1 and E2) function to control the degradation rate of the peptidic antigen (A), but may also serve any one or more additional functions. In some embodiments, the N- or C-terminal extensions (E1 or E2) may be free (one end of the N- or C-terminal extension is linked to the peptidic antigen (A) and the other end is not linked to another molecule) and may serve to slow the degradation of the peptidic antigen; for example, an E1 peptide-based extension may be linked to the N-terminus of the peptidic antigen by an amide bond to slow degradation. In other embodiments, the N- and / or C-terminal extensions (E1 and / or E2) may be linked to heterologous molecules and may function not only as linkers but also to regulate the degradation of the peptidic antigen (A). The N- and / or C-terminal extensions providing the linker function may link the peptidic antigen to a hydrophobic block (H) and / or a solubilizing block (S), such as PEG, either directly or indirectly via a linker U. In some embodiments, the extensions (E1 and / or E2) function to provide distance, i.e., space, 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 and / or C-terminal extensions (E1 and / or E2) can serve to stabilize particles formed by the peptide-antigen conjugate.

[0486] In some embodiments, the extensions (El and / or E2) contain charged functional groups, e.g., charged amino acid residues (e.g., arginine, ornithine, lysine, glutamic acid, aspartic acid, etc.), that confer a charge at pH 7.4. The number of charged residues present in the extensions can be used to modulate the net charge of the peptide-antigen conjugate. Peptide-based extensions (El and / or E2) that are recognized by proteases and confer specific electrostatic charges to stabilize particles formed by the peptide-antigen conjugates are described below.

[0487] Additionally, 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], where [ ] denotes that the group is optional. Thus, the amino acid sequence of the peptide-based E2 can be selected to disrupt peptide beta-sheet formation and prevent sequence truncation during solid-phase peptide synthesis. In a non-limiting example, the C-terminal dipeptide linker (E2) Gly-Ser is incorporated as a pseudoproline dipeptide during solid-phase peptide synthesis (e.g., Gly-Ser(Psi(Me,Me)pro)). In additional embodiments, proline is included in E2 (e.g., Ser-Pro-Leu-Arg (SEQ ID NO: 4)), where proline is included to facilitate production and promote processing of the extension by endosomal proteases.

[0488] In some embodiments, the peptide antigen (A) is linked at its C-terminus to an E2 extension that is linked either directly or indirectly via a linker (U) to a hydrophobic block, e.g., where the peptide antigen conjugate has the structure A-E2-UH or A-E2-H. In some embodiments, an E1 extension is linked to the N-terminus of the peptide antigen (A) and an E2 extension is linked at the C-terminus of the peptide antigen (A), where either E1 or E2 is linked to a hydrophobic block (H) either directly or via a linker (U), e.g., where 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 at its N-terminus to an E1 extension that is linked to a hydrophobic block (H) either directly or via a linker (U), e.g., where the peptide antigen conjugate has the structure HU-E1-A or H-E1-A. In some embodiments, a solubilizing block, such as PEG, is linked to extensions E1 or E2 that are linked to the N- or C-terminus of the peptide antigen (A), respectively, where the extension not linked to a solubilizing block (S), such as PEG, is linked to the hydrophobic block (H) either directly or via a linker (U), e.g., where 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.

[0489] In additional embodiments, PEG groups are linked to both the E1 and E2 extensions linked to both the N- and C-termini of the peptidic antigen (A), respectively, or PEG is linked to the E1 extension linked to the N-terminus of the peptidic antigen (A) but not to the E2 extension attached to the C-terminus of the peptidic antigen (A), which E2 extension may be linked to the hydrophobic block (H) either directly or via a linker (U). The linker precursor U1 or linker (U) may be linked to either extension (E1 or E2) by any suitable means, such as an amide bond.

[0490] In a preferred embodiment, the extensions (E1 and E2) are peptide sequences selected for recognition and hydrolysis by an enzyme, such as a protease. The extensions (E1 and E2) are preferably cleavable peptides that include amino acids recognized by either or both endosomal proteases and / or immunoproteasomes.

[0491] In some embodiments, the N-terminal extension (E1) is a peptide sequence about 1 to 8 amino acids in length, e.g., 1, 2, 3, 4, 5, 6, 7, or 8 amino acids, typically 10 amino acids or less, that is linked to the peptidic antigen (A) by an amide bond formed between the carboxyl group of E1 and the alpha amine of the N-terminal residue of peptidic antigen (A). The amide bond between E1 and peptidic antigen (A) can be enzymatically cleaved.

[0492] Amino acid positions are typically numbered from proximal to distal from the cleavage site, with amino acid positions C-terminal to the cleavage site indicated by a prime (e.g., Pn'). For example, in the case of a tetrapeptide extension (PN4-PN3-PN2-PN1) linked to the N-terminus of a peptide antigen (A), which 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 and PA1' is recognized and hydrolyzed by the enzyme.

[0493] In some embodiments, the N-terminal extension (E1) is an enzymatically degradable tetrapeptide recognized by endosomal proteases, wherein position PN1 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 (El) is an enzymatically degradable tripeptide recognized by an endosomal protease, wherein the PNl position of the tripeptide extension (e.g., PN3-PN2-PNl) 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 (El) is an enzymatically degradable dipeptide recognized by an endosomal protease, wherein the PNl position of the dipeptide extension (e.g., PN2-PNl) is preferably selected from arginine, lysine, citrulline, glutamine, threonine, leucine, norleucine, or methionine, and PN2 is selected from glycine, valine, leucine, or isoleucine. In yet a further embodiment, the N-terminal extension (E1) is an amino acid recognized by an endosomal protease, wherein the PN1 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 containing dipeptide is valine-citrulline.

[0494] 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 (e.g., PN4-PN3-PN2-Leu).

[0495] In additional embodiments, the N-terminal extension (El) is an enzymatically degradable peptide recognized by both endosomal proteases and immunoproteasomes, wherein positions PN5 and PN1 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 position PN5, recognized by cathepsins, and isoleucine, leucine, norleucine, or valine for position PN1, recognized by immunoproteasomes (e.g., PN8-PN7-PN6-Arg-PN4-PN3-PN2-Leu). A non-limiting example of an N-terminal extension (El) recognized by cathepsins and immunoproteasomes is Lys-Pro-Leu-Arg-Tyr-Leu-Leu-Leu (SEQ ID NO: 5).

[0496] 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, and Gly-Val-Ser. Non-limiting examples of dipeptide N-terminal extensions (E1) include Val-Cit, Val-Leu, Val-Arg, and 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 with Lys, Lys can be replaced with Arg, Glu can be replaced with Asp, and Asp can be replaced with Glu. Note that Cit = citrulline.

[0497] In some embodiments, E2 is a degradable peptide linked to the C-terminal residue of the peptide antigen (A) and contains an amino acid sequence that is recognized and hydrolyzed by a specific protease. In some embodiments, the C-terminal extension (E2) is a peptide sequence approximately 1 to 8 amino acids long, e.g., 1, 2, 3, 4, 5, 6, 7, or 8 amino acids, typically 10 amino acids or less. In a preferred embodiment, the C-terminal extension (E2) is linked 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 typically numbered from proximal to distal from the cleavage site, with amino acid positions C-terminal to the cleavage site indicated by a prime (e.g., Pn'). For example, in the case of 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 and PC1' is recognized and hydrolyzed by the enzyme.

[0498] In a preferred embodiment of the C-terminal extension (E2), the C-terminal extension (E2) comprises an amino acid sequence selected to promote immunoproteasome recognition and cleavage, and optionally endosomal protease recognition. Because peptide antigens (A) typically contain a C-terminal residue, e.g., leucine, that promotes immunoproteasome hydrolysis at the amide bond proximal to the C-terminal residue of peptide antigen (A), the extension linked to the C-terminus of peptide antigen (A) should be selected to promote immunoproteasome recognition and cleavage at the amide bond proximal to the C-terminus of peptide antigen (A). The immunoproteasome prefers a small, uncharged amino acid at the PC1' position adjacent to the C-terminal amino acid PA1 of peptide antigen (A), e.g., the amide bond between PA1 and 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). Thus, C-terminal extensions can be selected to facilitate recognition by either or both classes of proteases.

[0499] 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 between 1 and 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 a further embodiment, C-terminal extension E2 is a dipeptide PC1'-PC2', in which 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, C-terminal extension E2 is a tripeptide PC1'-PC2'-PC3', in which 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.

[0500] In a further embodiment, the C-terminal extension E2 is the tetrapeptide extension PC1'-PC2'-PC3'-PC4', in which 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 a further embodiment, the C-terminal extension E2 is the pentapeptide PC1'-PC2'-PC3'-PC4'-PC5', in which 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 a further embodiment, the C-terminal extension E2 is the 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.

[0501] 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), Gly-Gly-Glu-Leu-Val-Leu (SEQ ID NO: 24).

[0502] 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).

[0503] 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).

[0504] 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, and Gly-Pro-Cit. Non-limiting examples of dipeptide C-terminal extensions (E2) include Gly-Ser, Gly-Pro, Val-Cit, Gly-Arg, and 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 with Lys, Lys can be replaced with Arg, Glu can be replaced with Asp, and Asp can be replaced with Glu.

[0505] The C-terminal linker (E2) linked to the C-terminus of the peptide antigen (A) can be selected for recognition (i.e., hydrolysis) by both immune proteasomes 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 its C-terminus to a C-terminal tetrapeptide extension (E2) having the sequence PC1'-PC2'-PC3'-PC4', where 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 its C-terminus to a C-terminal hexapeptide extension (E2) having the sequence PC1'-PC2'-PC3'-PC4'-PC5'-PC6', where 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 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) linked at the C-terminus of peptide antigens (A) that favor 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).

[0506] Spacer (B) The spacer (B) is an optional component of the amphiphile that links the solubilizing block (S) to the hydrophobic block (H) either directly or via a linker (U), e.g., where the amphiphile has the structure SBH or SBUH. The spacer (B) may comprise any one or more of the following: an amino acid, including an unnatural amino acid; a hydrophilic polymer, e.g., an ethylene oxide-based polymer (PEG), an acrylate, methacrylate, acrylamide, or methacrylamide-based monomer; an alkane chain, or the like; or a combination thereof. The spacer (B) may be linked to the solubilizing block (S) and the hydrophobic block (H) by any suitable means, e.g., directly or indirectly via a linker, although the bond typically comprises a covalent bond, e.g., an amide bond.

[0507] In some embodiments, the spacer (B) functions to provide distance, i.e., space, between the heterologous molecules S and H. In other embodiments, the spacer (B) functions to impart hydrophobic or hydrophilic properties. In yet other embodiments, the spacer composition may be selected to impart rigidity or flexibility. In other embodiments, the spacer composition may be selected for recognition by enzymes to facilitate degradation.

[0508] In some embodiments, the spacer (B) is a hydrophilic polymer in which the monomer units are selected from acrylates, (meth)acrylates, acrylamides, (meth)acrylamides, allyl ethers, vinyl acetates, vinylamides, substituted styrenes, amino acids, acrylonitriles, heterocyclic monomers (e.g., ethylene oxide), sugars, phosphate esters, phosphonamides, sulfonate esters, sulfonamides, or combinations thereof.

[0509] In some embodiments, the spacer (B) is a peptide sequence of about 1 to 45 amino acids in length, e.g., 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 45 amino acids or less, linked to the hydrophobic block (H) and solubilizing block (S) by, for example, an amide bond formed between the N-terminus and C-terminal carboxyl group of the spacer (B), respectively. The amide bond between the spacer (B) and the solubilizing block (S) and / or the hydrophobic block (H) can be recognized by an enzyme or selected for resistance to enzyme-mediated hydrolysis.

[0510] In other embodiments, the spacer (B) is a hydrophilic polymer comprising monomer units selected from non-natural hydrophilic monomers, e.g., ethylene oxide (PEG), HPMA, poly(sarcosine), or HEMA, linked to the hydrophobic block (H) and the solubilizing block (S) either directly or via a linker, and having a length (i.e., degree of polymerization) of about 1 to 48 monomers, e.g., 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 48 monomers long or less.

[0511] Specific spacer compositions that result in unexpected improvements in biological activity are described throughout this specification. Note: Both the spacer group (B) and the solubilizing block (S) may comprise hydrophilic polymers (e.g., hydrophilic poly(amino acids); hydrophilic methacrylate-based polymers such as HEMA; hydrophilic methacrylamide-based polymers 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 an amphiphile. Similarly, the PEG group in 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.

[0512] Linker(U) The linker (U) optionally connects the solubilizing block (S) fragment (S-[B]-U1) to the hydrophobic block (H) fragment (U2-H) by reaction of U1 with U2 to form the amphiphile (S-[B]-UH).

[0513] The linker (U) also, independently of the amphiphile linker U, connects the peptide antigen conjugate fragment (PEG-[E1]-A-[E2]-U1 or U1-[E1]-A-[E2]-PEG) to the hydrophobic block (H) fragment (U2-H) by reaction of U1 with U2 to form the peptide antigen conjugate (PEG-[E1]-A-[E2]-UH or HU-[E1]-A-[E2]-PEG).

[0514] Although the peptide antigen (A) may be linked to the hydrophobic block (H) directly (i.e., A-H) or via an extension (i.e., A-E2-H (or H-E1-A)) entirely on the resin by solid-phase peptide synthesis, under certain circumstances it may be beneficial to generate the antigen (A) and the hydrophobic block (H) as separate fragments comprising the linker precursor U1 (PEG-[E1]-A-[E2]-U1 or U1-[E1]-A-[E2]-PEG) and the linker precursor U2 (U2-H), which may be linked on the resin or in solution to give PEG-[E1]-A-[E2]-UH (or HU-[E1]-A-[E2]-PEG).

[0515] Similarly, the solubilizing block (S) on the amphiphile may be linked 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, although under certain circumstances it may be advantageous to generate the solubilizing block (S) and the hydrophobic block (H) as separate fragments comprising the linker precursor U1 (S-[B]-U1) and the linker precursor U2 (U2-H), which may be linked on the resin or in solution to give S-[B]-UH.

[0516] In preferred embodiments, the linker precursor used to form the linker U is selected for site selectivity (i.e., reaction occurs only between U1 and U2, and not between other groups). In some embodiments, the linker precursor U1 comprises an activated carboxylic acid and is reacted with an amine-containing linker precursor U2 to form an amide-containing linker U, or U1 comprises an amine and is reacted with an activated carboxylic acid-containing U2 to form an amide-containing linker U. In some embodiments, the linker precursor U1 comprises a maleimide and is reacted with a thiol-containing linker precursor U2 to form a thioether-containing linker U, or U1 comprises a thiol and is reacted with a maleimide-containing U2 to form a thioether-containing linker U. In some embodiments, the linker precursor U1 comprises an azide and is reacted with an alkyne-containing linker precursor U2 to form a triazole-containing linker U, or U1 comprises an alkyne and is reacted with an azide-containing U2 to form a triazole-containing linker US.

[0517] In a preferred embodiment, amphiphiles of formula S-[B]-UH are joined together by linking a solubilizing block fragment (S-[B]-U1) to a hydrophobic block fragment (U2-H), where the linker precursor U1 contains a strained alkyne (e.g., dibenzocyclooctyne (DBCO), bicyclononyne (BCN), etc.), which is reacted with an azide-containing linker precursor U2 to form a triazole-containing linker U.

[0518] In a preferred embodiment, peptide antigen conjugates of the formula PEG-[E1]-A-[E2]-UH or HU-[E1]-A-[E2]-PEG are 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), where the linker precursor U1 contains a strained alkyne (e.g., dibenzocyclooctyne (DBCO), bicyclononyne (BCN), etc.), which is reacted with an azide-containing linker precursor U2 to form a triazole-containing linker U.

[0519] In a preferred method, peptide antigen conjugates of the formula PEG-[E1]-A-[E2]-UH or HU-[E1]-A-[E2]-PEG are prepared by (i) adding 1 molar equivalent of peptide antigen fragment in DMSO at a concentration greater than 10 mM, preferably greater than 25 mM, to at least 1.05 equivalents of hydrophobic blocking fragment in DMSO at a concentration greater than 25 mM, most preferably greater than 50 mM, and (ii) adding an azide resin, such as agarose-azide, to the reaction mixture upon completion of the reaction. The peptide antigen fragments PEG-[E1]-A-[E2]-U1 or U1-[E1]-A-[E2]-PEG are linked together by linking them to the hydrophobic block fragment (U2-H) by adding HCl to remove any unreacted hydrophobic block fragments, and then (iii) removing the resin to yield the pure peptide antigen conjugate, where the linker precursor U1 contains DBCO and the linker precursor U2 contains azide.

[0520] In another preferred embodiment, a linker precursor U1 contains an azide, which is reacted with a linker precursor U2 containing a strained alkyne (e.g., dibenzocyclooctyne (DBCO), bicyclononyne (BCN), etc.) to form a triazole-containing linker U. In a non-limiting example, a linker precursor U2 comprising DBCO is linked to a 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 a solubilizing block fragment (S-[B]-U1) or a peptide antigen fragment (PEG-[E1]-A-[E2]-U1 or U1-[E1]-A-[E2]-PEG) via a suitable linker X.

[0521] In a preferred embodiment, the linker U preferably comprises an amide, a thioether or a triazole.

[0522] Dendron Amplifiers A dendron amplifier is a specific type of linker moiety that functions to increase the valency (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 a solubilizing block (S), a dendron amplifier is used to increase the valency of the solubilizing groups (referred to herein as "SG") present on the surface of the solubilizing block (S). In other embodiments, a dendron amplifier is used to increase the valency of the solubilizing block (S) and spacer (B) linked to the hydrophobic block (H).

[0523] Dendron amplifiers (also referred to as "dendrons") are regularly branched molecules that are often symmetrical and typically comprise repeating monomeric units containing three or more functional groups (FG) and branch points. Dendron amplifiers can be represented by the formula (FG')-T-(FGt)d, where FG' and FGt are focal 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 2 to 32, but more preferably 2 to 8, e.g., 2, 3, 4, 5, 6, 7, and 8. The factor by which the dendron amplifier increases the terminal functional group (FGt) is FGt=β γ where β is the number of branches that occur per dendron generation, and the symbol γ is the generation number, where the number of branches can be any integer, but typically between 2 and 6, and the number of generations can be any integer, but typically between 1 and 10. The free (i.e., unreacted) terminal functional groups present on the solubilizing block can also be referred to as solubilizing groups (SG).

[0524] A dendron amplification material may comprise repeating monomers comprising a first functional group (FG1) and a second functional group (FG2), where the first functional group is reactive to a second functional group. For example, a non-limiting example of a second generation dendron amplification material having β=2 is shown here for clarity, comprising repeating monomers comprising a first functional group (FG1) and a second functional group (FG2), where the first functional group is reactive to a second functional group: [ka] In the formula, the first functional group at the starting point is also referred to as the focal functional group (FG') and the terminal FG2 is referred to as the terminal functional group or FGt.

[0525] A non-limiting example of a third generation dendron formed from a monomer containing a first functional group and a second functional group, where β=2, is shown here for clarity. [ka]

[0526] A non-limiting example of a second generation dendron amplifier having β=3, comprising a repeating first monomer comprising a first functional group (FG1) and a second functional group (FG2), where the first functional group is reactive towards the second functional group, is shown here for clarity. [ka]

[0527] A monomer comprising a first functional group and a second functional group, where the first functional group is reactive to the second functional group, the monomer comprising at least one first functional group and two or more second functional groups can be selected from any suitable monomer. 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, wherein R 1 are independent, (CH2) y3 -FG2, (OCH2CH2) y3 -FG2 or CH2(OCH2CH2) y3 -FG2), and y2 and y3 are each a repeating unit of an integer selected from 1 to 6.

[0528] FG1-CH(R 1 )2 (wherein FG1 is NH2 and R 1 is CH2(OCH2CH2) y3 -FG2, y3 is 1, and FG2 is COOH) are shown here for clarity: [ka]

[0529] wherein the above monomers are used to generate a second generation amplification linker, the structure of which is: [ka]

[0530] Additional non-limiting examples of monomers comprising a first functional group and a second functional group, where the first functional group is reactive to the second functional group, include FG1-(CH2) y2 N(R 2 )2, FG1-(CH2CH2O) y2 CH2CH2N(R 2 ) 2, wherein R 2 are independent, (CH2) y3 -FG2, (CH2CH2O) y3 (CH2) y4 -FG2, (CH2OCH2CH2) y3 -FG2), where y2, y3, and y4 are each repeating units that are integers selected from 1 to 6. Note: In the above example, FG' is an amine, and the four FGt are carboxylic acids.

[0531] FG1-(CH2CH2O) y1 CH2CH2N(R 2 )2 (wherein FG1 is NH2 and R 2 is (CH2CH2O) y3 (CH2) y4 A non-limiting example of -FG2, y2 is 2, y3 is 1, y4 is 2, and FG2 is COOH) is provided here for clarity. [ka]

[0532] Still additional non-limiting examples of monomers comprising a first functional group and a second functional group, where the first functional group is reactive to the second functional group, that comprise 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. A non-limiting example of a third generation lysine dendron is provided here for clarity. [ka]

[0533] A dendron amplification material may comprise two repeating monomers, where the first monomer comprises three or more first functional groups (FG1) and the second monomer comprises two or more second functional groups (FG2), and the first functional groups are reactive to the second functional groups. For example, a non-limiting example of a second generation dendron amplification material having β=2, comprising repeating first and second monomers, where the first monomer comprises three first functional groups (FG1) and the second monomer comprises two second functional groups (FG2), and the first functional groups are reactive to the second functional groups, is provided here for clarity. [ka]

[0534] A non-limiting example of a first generation dendron amplifier having β=2, comprising a repeat of a first monomer and a second monomer, where the first monomer comprises three first functional groups (FG1) and the second monomer comprises three second functional groups (FG2), and the first functional groups are reactive towards the second functional groups, is shown here for clarity. [ka]

[0535] Dendron amplifiers can be used to link together any three or more components of amphiphiles, peptide-antigen conjugates, and drug molecule conjugates. The focal functional group (FG') and terminal functional group (FGt) can be further functionalized, i.e., reacted, to suit a particular purpose.

[0536] In a preferred embodiment of an amphiphile of formula S-[B]-[U]-H, the solubilizing block (S) comprises a dendron amplifier, where the focal point is linked to the hydrophobic block (H) either directly or indirectly via a spacer (B) and / or a linker U, and the terminal functional group (FGt) is either unlinked, serves as a solubilizing group, or is linked to a solubilizing group (SG). The solubilizing group (SG) is any molecule that is hydrophilic and / or charged, and preferred solubilizing groups (SG) are described throughout this specification.

[0537] In some embodiments of amphiphiles of formula S-[B]-[U]-HD, peptide antigen conjugates of formula PEG-[E1]-A-[E2]-[U]-HD or HD-[U]-[E1]-A-[E2]-PEG, and drug molecule conjugates of formula HD, the hydrophobic block (H) comprises a dendron amplifier, where the focal point is either (i) linked to the solubilizing block (S) either directly or indirectly via a spacer (B) and / or a linker U, (ii) linked to the antigen (A) either directly or indirectly via an extension (E1 or E2) and / or a linker U, or (iii) linked to the drug molecule either directly or via a linker X1.

[0538] In some embodiments, the hydrophobic block (H) comprises a dendron amplifier, and the terminal functional group (FGt) is linked to a hydrophobic drug molecule. In such embodiments, the focal point is either (i) linked to the solubilizing block (S) either directly or indirectly via a spacer (B) and / or a linker U, (ii) linked to the antigen (A) either directly or indirectly via an extension (E1 or E2) and / or a linker U, or (iii) unreacted or capped with a terminal group such as an acetyl group. Capping refers to the modification of 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 may be capped with an activated carboxylic acid (e.g., acetyl chloride) to give a less reactive amide, or, for example, a strained alkyne may be capped with an alkyl-azide to give a less reactive triazole. Hydrophobic block (H)

[0539] The hydrophobic block (sometimes designated "H" in the formula) is a molecule with substantially limited aqueous solubility or is amphiphilic in character and can organize 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 at a certain concentration, temperature, and / or pH range, but becomes insoluble in response to changes in concentration, temperature, and / or pH. For example, in some embodiments, the hydrophobic block is a hydrophobic polymer that is temperature-responsive, i.e., the hydrophobic polymer has a transition temperature (T tr), but becomes insoluble above the transition temperature. Preferred hydrophobic blocks (H) are molecules that have a solubility of at least less than about 1.0 mg / mL, e.g., less than about 0.1 mg / mL or less than about 0.01 mg / mL, at or near physiological pH (about pH 7.4), about pH 6.5 to pH 8.5, or about pH 6.0 to pH 9.0, and at or near physiological temperature (about 37°C) and physiological salt concentrations (about 10 g / L) and salt compositions.

[0540] The hydrophobic block (H) may be selected from any molecule including higher alkanes, cyclic aromatic compounds, fatty acids, terpenes / isoprene-derived compounds, or polymers or oligomers with limited water solubility and / or amphiphilic properties.

[0541] 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, e.g., myristic acid. In other embodiments, the hydrophobic block (H) comprises a diacyl lipid, such as 1,2-dioleyl-sn-glycero-3-phosphoethanolamine or 1,2-distearoyl-sn-glycero-3-phosphoethanolamine, or a lipopeptide, e.g., 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, such as QS-21.

[0542] In some embodiments, the hydrophobic block (H) is a linear, branched, or brush polymer (or oligomer). The hydrophobic block (H) can be a homopolymer or a copolymer. The hydrophobic block (H) can contain one or many different types of monomer units. The hydrophobic block (H) can be a statistical copolymer or an alternating copolymer. The hydrophobic block (H) can be a block copolymer, such as an AB type, or the polymer can comprise a graft copolymer, in which two or more polymers are linked by a polymer-analogous reaction.

[0543] The hydrophobic block (H) may comprise polymers containing naturally occurring and / or non-naturally occurring monomers and combinations thereof.

[0544] In some embodiments, the hydrophobic block (H) is selected from naturally occurring biopolymers. Naturally occurring biopolymers can include peptides containing hydrophobic amino acids (sometimes referred to as poly(amino acids)). Non-limiting examples of hydrophobic amino acids include leucine, isoleucine, norleucine, valine, tryptophan, phenylamine, tyrosine, and methionine, as well as hydrophilic amino acids that have been modified, such as by acetylation or benzoylation, to have hydrophobic properties. Naturally occurring biopolymers that are water-soluble in their natural form may be used, but must be chemically modified to render them water-insoluble and suitable for use as the hydrophobic block (H). For example, biopolymers composed of hydrophilic amino acids such as glutamic acid or lysine residues may be modified with gamma carboxyl or epsilon amine groups, respectively, to increase the hydrophobicity of the resulting modified biopolymer for attachment of hydrophobic molecules, such as hydrophobic drug molecules. Similarly, the biopolymer may be selected from hydrophilic polysaccharides, which may include, but are not limited to, glycogen, cellulose, dextran, alginate, and chitosan, provided that such polysaccharides are chemically modified, e.g., via acetylation or benzoylation of hydrophilic functional groups, to render the resulting modified polysaccharide water-insoluble. In yet a further embodiment, the hydrophobic block comprises a monomer selected from lactic acid and / or glycolic acid.

[0545] The monomers constituting the hydrophobic block (H) may be selected from acrylates, (meth)acrylates, acrylamides, (meth)acrylamides, allyl ethers, vinyl acetates, vinylamides, substituted styrenes, amino acids, acrylonitriles, heterocyclic monomers (e.g., ethylene oxide), sugars, phosphate esters, phosphonamides, sulfonate esters, sulfonamides, or combinations thereof. Specific examples of (meth)acrylates and (meth)acrylamides include benzyl methacrylamide (BnMAM) and benzyl methacrylate (BnMA), respectively.

[0546] Certain monomers described herein as hydrophobic monomers may be water-soluble under certain conditions, but are hydrophobic and water-insoluble under certain conditions in aqueous solution. Non-limiting examples include temperature-responsive monomers such as N-isopropylmethacrylamide (NIPMAM); homopolymers composed entirely of NIPMAM may be water-soluble at room temperature, but become insoluble and form particles at elevated temperatures. This distinction is made to facilitate the description of certain embodiments. In some embodiments, the hydrophobic block is composed predominantly of monomer units selected from NIPAM, NIPMAM, N,N'-diethylacrylamide (DEAAM), N-(L)-(1-hydroxymethyl)propylmethacrylamide (HMPMAM), N,N'-dimethylaminoethyl methacrylate (DMEMA), N-(N-ethylcarbamido)propylmethacrylamide, N-vinylisobutyramide (PNVIBA), N-vinyl-n-butylamide (PNVBA), N-acryloyl-N-propylpiperazine (PNANPP), N-vinylcaprolactam (PVCa), DEGMA, TEGMA, or hydrophobic monomers that are temperature responsive (sometimes referred to as "temperature responsive monomers"), such as poly(amino acid) or γ-(2-methoxyethoxy)esteryl-L-glutamate. In still other embodiments, the hydrophobic block (H) may comprise monomers of ethylene oxide, propylene oxide, or combinations thereof.

[0547] The hydrophobic block (H) comprising a polymer typically comprises a hydrophobic monomer and one or more other types of monomers, such as a reactive monomer, a spacer monomer, and / or a charged monomer, optionally linked to a drug molecule. In some embodiments of the hydrophobic block (H) comprising a polymer (or oligomer), the majority of the monomer units are selected from hydrophobic monomers. In other embodiments of the hydrophobic block (H) comprising a polymer (or oligomer), the majority of the monomer units are selected from reactive monomers linked to hydrophobic drug molecules. In yet 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 yet further embodiments of the hydrophobic block (H) comprising a polymer (or oligomer), the polymer comprises a hydrophobic monomer and a charged monomer, and optionally a reactive monomer linked to a hydrophobic drug molecule.

[0548] In a preferred embodiment, the hydrophobic block (H) comprises a polymer (or oligomer) comprising a hydrophobic monomer further comprising an aryl group. In certain embodiments, the hydrophobic block (H) comprises a heteroaryl group. In yet other embodiments, the aryl or heteroaryl group of the hydrophobic block (H) comprises an amino substituent. The inventors have found that hydrophobic blocks (H) comprising aminoaryl or aminoheteroaryl groups provide improved manufacturability and solubility in water-miscible solvents. The inventors have also found that amphiphiles having hydrophobic blocks (H) comprising aromatic amines provide for the formation of stable particles with low CMCs.

[0549] 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 quinolinyl. Non-limiting examples include: [ka] [ka] [ka] where X is any suitable linker molecule and y is an integer value, typically 1-6.

[0550] In a preferred embodiment, the aryl or heteroaryl group includes: [ka] These include, but are not limited to:

[0551] Furthermore, one or more hydrogen atoms in the aforementioned aryl or heteroaryl groups may be substituted with one or more fluorine atoms. In certain embodiments, the hydrophobic block comprises a fluorinated aliphatic group, aryl group, or heteroaryl group, wherein one or more hydrogen atoms in the aforementioned groups comprising the hydrophobic monomer may be substituted with one or more fluorine atoms. The following non-limiting examples of fluorinated aryl groups may be present in the hydrophobic monomer: [ka] where X is any suitable linker molecule and y is an integer value, typically from 1 to 6.

[0552] The present inventors have unexpectedly discovered that hydrophobic blocks (H) comprising aminoaryl or aminoheteroaryl groups provide 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, where Ar can be aryl or heteroaryl, and R is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkyl, heteroaryl, or heteroarylalkyl. Non-limiting examples of aminoaryl or aminoheteroaryl groups include: [ka] where X is any suitable linker molecule and y is an integer value, typically from 1 to 6.

[0553] In some embodiments, the hydrophobic block (H) comprises a polymer (or oligomer) further comprising a hydrophobic monomer having a fused aryl group (e.g., naphthyl) or a fused heteroaryl group (e.g., xanthenyl 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 therefore, a reactive monomer linked to a hydrophobic drug molecule comprising an aromatic group can also be described as a hydrophobic monomer or a reactive monomer linked to a drug comprising an aromatic group.

[0554] In some embodiments, the hydrophobic block (H) has formula I: [ka] wherein the poly(amino acid) of Formula I comprises monomers selected from hydrophobic amino acids (M), reactive amino acids (N), spacer amino acids (O), charged amino acids (P), and combinations thereof, provided that at least monomer M or N is present, and m, n, o, and p represent the presence of an integer number of repeating units of monomers M, N, O, and P, respectively, which may be distributed along the polymer in a specified or random order, and R is typically hydrogen, NH, NH—CH, NH—(CH) y5 selected from CH3, OH, or a drug molecule (D) linked either directly or via X1.

[0555] In some embodiments, P is absent. In other embodiments, N, O, and P are each absent.

[0556] In some embodiments, P is [ka] where each R 5 are independently a group containing 1 to 2 charged functional groups.

[0557] In some embodiments, O is [ka] wherein each Q is independently (CH) y6 and (CH2CH2O) y7 CH2CH2, each y6 is independently selected from an integer of 1 to 6, and each y7 is independently selected from an integer of 1 to 4.

[0558] In some embodiments, N is [ka] wherein each X1 is independently a suitable linker and each D is independently a drug molecule.

[0559] In some embodiments, M is [ka] where each R 4 are independently hydrophobic groups.

[0560] In some embodiments, the hydrophobic block (H) has formula I: [ka] wherein the poly(amino acid) of formula I comprises monomers selected from hydrophobic amino acids (M), reactive amino acids (N), spacer amino acids (O), charged amino acids (P), and combinations thereof, provided that at least monomer M or N is present, and m, n, o, and p represent the presence of an integer number of repeating units of monomers M, N, O, and P, respectively, which may be distributed along the polymer in a specified or random order; R 3 is typically hydrogen, NH2, NH2-CH3, NH2-(CH2) y5 CH3, OH, or a drug molecule (D) linked either directly or via X1; R 4 is any hydrophobic group, typically selected from an aryl or heteroaryl group, and R 5 is any group that is charged in aqueous solution or is pH-responsive and contains one or more functional groups that are charged in aqueous solution over a certain pH range, and Q is typically (CH2) y6 and (CH2CH2O) y7The N-terminus is selected from any lower alkyl or heteroalkyl, including, but not limited to, CH2CH2 (where y6 is any integer between 1 and 6, and y7 is typically an integer selected from 1 to 4), and is either (i) linked directly to the solubilizing block (S) or indirectly via a spacer (B) and / or a linker U, (ii) linked to the peptide antigen (A) either directly or indirectly via an extension (E1 or E2) and / or a linker U, or (iii) linked to a drug molecule either directly or via X1. Note: Hydrophobic amino acids, reactive amino acids, spacer amino acids, and charged amino acids are sometimes more commonly described as hydrophobic monomers, reactive monomers, spacer monomers, and charged monomers, respectively.

[0561] In a preferred embodiment of the poly(amino acid) of formula I, R 4 teeth, [ka] where: a is aryl or heteroaryl; X2 is present or absent and, if present, is a suitable linker; y8 is selected from integers from 0 to 6; Z 1 , Z 2 , and Z 3 are each independently selected from H, F, hydroxy, amino, alkyl, and fluoroalkyl.

[0562] In preferred embodiments of the poly(amino acid) of Formula I, α is aryl, such as phenyl or naphthyl. In other embodiments, α is heteroaryl, such as imidazolyl, pyridinyl, quinolinyl, isoquinolinyl, indolyl, and benzimidazolyl.

[0563] In preferred embodiments of the poly(amino acid) of Formula I, X2 is absent. In other embodiments, X2 is present and is selected from the group consisting of C(O), CO2(CH2),y9 , and C(O)NH(CH2) y9 , NHC(O) and NHC(O)(CH2) y9 where y9 is an integer typically selected from 1 to 6. In other embodiments, X2 is present and is selected from a lower alkyl and a PEG group.

[0564] In a preferred embodiment of the poly(amino acid) of Formula I, the poly(amino acid) of Formula I does not contain a hydrophobic group R 4 In a preferred embodiment, R 4 is selected from hydrophobic groups including aryl groups, heteroaryl groups, aminoaryl groups, and / or aminoheteroaryl groups. 4 Non-limiting examples of [ka] [ka] [ka] [ka] wherein X2 is any suitable linker molecule and y8 is an integer value, typically from 0 to 6. In a preferred embodiment, y8 is 1.

[0565] In a non-limiting example, R 4 but, [ka] and the monomer M is [ka] is.

[0566] 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, and a drug molecule (D) is linked to the monomer directly or via X. Suitable reactive amino acids include any amino acid bearing a group suitable for attachment of a drug molecule, including, but not limited to, azide, alkyne, tetrazine, transcyclooctyne (TCO), protected hydrazine, ketone, aldehyde, certain hydroxyl groups, isocyanate, isothiocyanate, carboxylic acid, activated carboxylic acid, activated carbamate, activated carbamate, protected maleimide, thiol, and / or amine groups.

[0567] X1 is any suitable linker for linking the drug molecule D to the hydrophobic block (H), including linking to a reactive amino acid N of a poly(amino acid), and is typically -(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 linked at the N-terminus or the amine group is removed), or -NH-(CH) y10 -FG3 and -NH-(CH2) y10 -R 6 (when the drug is linked at the C-terminus or the carbonyl group is removed), where y is an integer typically selected from 1 to 6, and R 6 is typically -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, wherein R 7 is typically -(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-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 typically selected from 1 to 6; 8is any amino acid side chain, and W can be independently selected from H (hydrogen), FG3, LG, and w, where FG3 can be selected from, but is not limited to, carboxylic acids, activated carboxylic acids (e.g., carbonylthiazolidine-2-thione ("TT"), NHS, or nitrophenol esters), carboxylic acid anhydrides, amines and protected amines (e.g., tert-butyloxycarbonyl-protected amines), OSi(CH3), alkenes, azides, alkynes, dye alkynes, halogens (e.g., fluoride, chloride), olefins and endocyclic olefins (e.g., allyl), CN, OH, and epoxy, hydrazines (including hydrazides), carbohydrazides, aldehydes, ketones, carbamates, and activated carbamates. , LG is any suitable leaving group which may be selected from, but is not limited to, any suitable leaving group (e.g., NHS, TT, nitrophenol, etc.); w results from either the reaction of FG4 with FG3 or the displacement of LG with FG4 and is typically a group 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)-, where w is always linked to D either directly (i.e., w-D) or indirectly via X3 (i.e., w-X3-D).

[0568] Drug molecule (D) may be attached to reactive amino acid N directly or through X1 by reaction of FG4 with FG3, where FG4 is any suitable functional group on drug (D) that is reactive with FG3. Alternatively, drug molecule (D) may be linked to reactive amino acid N through X1 by substitution of LG with any suitable FG4 that contains a nucleophile, e.g., a primary amine, or drug molecule (D) may be linked to reactive amino acid N through X1 by substitution of LG present on the drug molecule with any suitable FG3 that contains a nucleophile.

[0569] 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 -(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), indicating that the drug is linked via an amide bond at the carbonyl of X1, which (after amide bond formation) is -(CH2) y10 -R 6 where y10 is 2 and R 6 =C(O)-W, where W is a group w that is NH-, which is linked to D to give -(CH2)2-C(O)-NH-D.

[0570] The drug may additionally comprise a linker X3 between the reactive functional group FG4 and the pharmacophore (e.g., FG4-X3-D). Specific preferred compositions of X3 are described elsewhere.

[0571] 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(O)-D (carbonyl shown), indicating that the drug is linked at the amine of X1 via an amide bond.

[0572] In still other embodiments, FG3 is a ketone or aldehyde and FG4 is a hydrazide or carbohydrazide, which react to form a hydrazone. In a non-limiting example, X1 is -(CH2) y10 -R 6, y10 is 4, and R 6 is -NH-C(O)-R 7 and R 7 is (CH2) y11 -W, y11 is 2, W is C(O)-CH3, and FG4 present on the drug molecule is a hydrazide (NH2-NH2-C(O)-D), which reacts with X1, i.e., -(CH2)4-NH-C(O)-(CH2)2-C(O)-CH3, to form a hydrazone bond, i.e., -(CH2)4-NH-C(O)-(CH2)2-C(CH3)=N-NH-C(O)-D. In yet other embodiments, FG3 is a hydrazide or carbohydrazide, and FG4 is a ketone or aldehyde, which react to form a hydrazone. In a non-limiting example, X1 is -(CH2) y10 -R 6 , y10 is 2, R 6 is -C(O)-W, W is FG3, FG3 is -NH-NH2, and FG4 present on the drug molecule is a ketone C...

Claims

1. 1. 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: A is a peptide antigen, E1 is the N-terminal extension, E2 is a C-terminal extension, H, independently at each occurrence, is a hydrophobic block; and one or more drug molecules (D) are optionally attached to each H, either directly or via a suitable linker, X1; U is, independently at each occurrence, a linker; [ ] means that the group is optional; The above vaccine, wherein - indicates that two adjacent groups are directly linked to each other by a covalent bond, or indirectly linked to each other via a suitable linker X.

2. the vaccine further comprises an amphiphile having the formula S-[B]-[U]-H, where S is a solubilizing block; B is a spacer, H is a hydrophobic block; U is a linker, [ ] means that the group is optional; - indicates that two adjacent groups are directly linked to each other by a covalent bond or indirectly linked to each other via a suitable linker X; 2. The vaccine of claim 1, wherein the amphiphile S comprises a dendron amplifier.

3. 2. The vaccine of claim 1, wherein the PEG group of the peptide antigen conjugate comprises 12 to 36 monomer units.

4. E1 is present, i) a single amino acid PN1 selected from arginine, lysine, citrulline, glutamine, threonine, leucine, norleucine, and methionine; ii) dipeptides PN2-PN1, where PN1 is selected from arginine, lysine, citrulline, glutamine, threonine, leucine, norleucine, and methionine, and PN2 is selected from glycine, valine, leucine, and isoleucine; iii) the tripeptide PN3-PN2-PN1, where 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, where 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; 2. The vaccine of claim 1, comprising an enzyme-degradable peptide sequence comprising:

5. E2 is present, i) a single amino acid PC1′ selected from glycine, serine, arginine, lysine, citrulline, glutamine, threonine, leucine, norleucine and methionine; ii) dipeptides PC1'-PC2', where 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) the tripeptide PC1'-PC2'-PC3', where 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) the tetrapeptide PC1'-PC2'-PC3'-PC4', where 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) the 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; 2. The vaccine of claim 1, comprising an enzyme-degradable peptide sequence comprising:

6. 3. The vaccine of claim 2, wherein the S of the amphiphile comprises two or more solubilizing groups (SG).

7. 7. The vaccine of claim 6, wherein the two or more SGs are connected to the remainder of the S by a dendron amplifier.

8. 8. The vaccine of any one of claims 6 to 7, wherein the SGs are independently selected from amines, hydroxyls, carboxylic acids and / or sugar molecules, and the sugar molecules are independently selected from mannose, glucose, glucosamine, N-acetylglucose, galactose, galactosamine, and N-acetylgalactosamine, N-acetylglucosamine, phosphoserine and any derivatives thereof, an agonist of CD22a, sialyl Lewis x, and combinations thereof.

9. The vaccine of any one of claims 7 to 8, wherein the dendron amplifier comprises 1 to 10 generations of repeating monomer units with 2 to 6 branches per generation.

10. The repeating monomer unit is FG1-(CH ２ ) ｙ２ CH (R １ ) ２ , FG1-(CH ２ ) ｙ２ C (R １ ) ３ , FG1-(CH ２ CH ２ O) ｙ２ CH (R １ ) ２ , FG1-(CH ２ CH ２ O) ｙ２ C (R １ ) ３ , and FG1-CH(R １ ) ２ , FG1-C(R １ ) ３ is selected from During the ceremony, R １ is calculated independently for each occurrence as follows: (CH ２ ) ｙ３ -FG2, (OCH ２ CH ２ ) ｙ３ -FG2, and CH ２ (OCH ２ CH ２ ) ｙ３ -FG2), y2 and y3 are independently at each occurrence a repeat unit that is an integer from 1 to 6; FG1 is a first functional group, 10. The vaccine of claim 9, wherein FG2 is the second functional group.

11. FG1 is -NH ２ and FG2 is -CO ２ H or FG1 is —CO ２ H, and FG2 is —NH ２ The vaccine of claim 10, wherein

12. The vaccine of any one of claims 7 to 11, wherein the dendron amplifier comprises a polyethylene oxide (PEG) group.

13. 13. The vaccine of 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. 14. The vaccine of 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. 15. The vaccine of claim 13 or 14, wherein each H independently comprises a poly(amino acid) comprising monomers selected from hydrophobic amino acids (M), reactive amino acids (N), spacer amino acids (O), charged amino acids (P), and combinations thereof, provided that at least one of M or N is present.

16. Each H independently represents the formula: 【Chemistry 1】 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 of 1 to 100, and the sum of m, n, o, and p is 100 or less; R ３ is hydrogen, NH ２ , NH-CH ３ , NH-(CH ２ ) ｙ５ CH ３ , OH, or a drug molecule (D) attached either directly or via a suitable linker X; The vaccine of claim 15, wherein y5 is an integer selected from 1 to 6.

17. P, if present, 【Chemistry 2】 wherein each R ５ The vaccine of claim 16, wherein each is independently a group containing 1 to 2 charged functional groups.

18. O, if present, 【Transformation 3】 wherein each Q is independently (CH ２ ) ｙ６ and (CH ２ CH ２ O) ｙ７ CH ２ CH ２ wherein 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. N, if present, 【Chemistry 4】 wherein each X1 is independently a suitable linker and each D is independently a drug molecule.

20. M, if present, 【Transformation 5】 wherein each R ４ The vaccine of any one of claims 16 to 19, wherein are independently a hydrophobic group.

21. R ４ but, 【Transformation 6】 where: a is aryl or heteroaryl; X2 is present or absent and, if present, is a suitable linker; y8 is selected from integers from 0 to 6; Z １ , Z ２ , and Z ３ 21. The vaccine of claim 20, wherein each is independently selected from hydrogen, fluorine, hydroxy, amino, alkyl, and fluoroalkyl.

22. Each R ４ became independent, 【Transformation 7】 wherein each X2 is a direct linkage or is independently selected from a suitable linker, and each y8 is independently selected from an integer from 0 to 6.

23. 23. The vaccine of 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. 24. The vaccine of any one of claims 2 to 23, wherein B, if present, is a hydrophilic polymer or peptide.

25. 25. The vaccine of any one of claims 2 to 24, wherein the U of the amphiphile, when present, comprises an amide, a thioether, or a triazole.

26. 25. The vaccine of any one of claims 1 to 24, wherein U of the peptide antigen conjugate, when present, comprises an amide, a thioether, or a triazole.

27. 27. The vaccine of any one of claims 1 to 26, wherein the vaccine comprises a molar ratio of peptide antigen conjugate to amphiphile of about 4:1 to about 1:20, preferably 1:

1.

28. The vaccine of any one of claims 1 to 27, wherein the vaccine is a tolerogenic allergy vaccine, a tolerogenic autoimmune disease vaccine, or a tolerogenic transplant rejection vaccine.

29. 28. A method of treating or preventing an autoimmune disease, allergy or infectious disease in a subject in need thereof, the method comprising administering to the subject a vaccine according to any one of claims 1 to 27.

30. 28. The vaccine of any one of claims 1 to 27, wherein the at least one peptide antigen conjugate comprises (A) selected from an autoantigen, an alloantigen and an allergen, comprising a sequence of 7 to 45 amino acids in length.

31. The vaccine of any one of claims 1 to 27, wherein at least one peptide antigen (A) comprises alpha aminobutyric acid and / or norleucine.

32. A is 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), QQFLQPEQPFPQQPEQPEQPYPQQPEQPFPQPQQ (SEQ ID NO: 490), QQFSQPEQEFPQPQQPQQSFPEQQPPF (SEQ ID NO: 491), PTPLQPEQPFPQQP 28. The vaccine of any one of claims 1 to 27, selected from the group consisting of QQPQQPFPQPEQPFPWQPQ (SEQ ID 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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