Self-assembled nanoparticles based on amphiphilic peptides

Abstract

The present disclosure relates to a vaccine comprising an amphiphile having the formula S-[B]-[U]-H and at least one peptide antigen conjugate having a formula selected from [S]-[E1]-A-[E2]-[U]-H and H-[U]-[E1]-A-[E2]-[S], wherein the amphiphile and / or the at least one peptide antigen conjugate comprises a dendron amplifier. The vaccine is useful for treating or preventing cancer, autoimmune disease, allergy, infectious disease, cardiovascular disease, or neurodegenerative disease.

Description

[Technical Field]

[0001] This application claims the benefit of priority to U.S. Provisional Patent Application No. 63 / 197,092, filed June 4, 2021, and U.S. Provisional Patent Application No. 63 / 149,996, filed February 16, 2021, each of which is hereby incorporated by reference in its entirety.

[0002] This invention was made in fulfillment of a Cooperative Research and Development Agreement with the U.S. Department of Health and Human Services, National Institutes of Health. The U.S. Government has certain rights in this invention.

[0003] Technical Field The present disclosure relates to novel amphiphilic compositions, particularly peptide amphiphilic compositions, that can be used to form nanoparticles, including micellar structures or polymersomes, to methods of making the amphiphilic compositions, to processes for formulating drug molecules with the amphiphilic compositions to form nanoparticles, and to therapeutic uses of the nanoparticles for drug delivery. [Background technology]

[0004] background Vaccines can be used to generate antibody and / or T cell responses against any pathogen or molecule associated with a disease as a means of preventing disease onset or reducing disease severity. The choice of vaccination platform utilized will depend in part on the target pathogen or molecule and the type of immune response required.

[0005] Peptide-based vaccines include synthetic peptide antigens derived from any protein associated with an infectious organism, toxin, or pathology (e.g., prions, misfolded proteins, etc.), which are used to induce antibody and T cell responses against peptide antigens that cross-react with natural proteins present on the infectious organism, toxins, or multiple types of disease-related proteins. Because peptides alone are not immunogenic, peptide antigens are typically formulated with immune stimulants and / or delivery vehicles called vaccine adjuvants. A major advantage of peptide-based vaccines is that small fragments of pathogen-derived or disease-causing proteins, including those with post-translational modifications (e.g., glycans, phosphate groups, etc.), can be synthetically produced and used as antigens to induce a highly focused immune response against the conserved structures (i.e., peptide sequences and any higher-order structures) of pathogens, toxins, and disease-causing proteins. The benefit of strictly targeting the immune response to a specific site of interest is that it avoids off-target antibody and T cell responses that could weaken the immune response and potentially lead to pathology.

[0006] Based on these characteristics, peptide-based vaccines are particularly well suited to focusing T cell responses against tumor antigens to prevent or treat cancer. Therefore, tumor-derived peptide antigens can be used to focus immune responses against cancer cells while avoiding immune responses against healthy tissues. Similarly, peptide-based vaccines are particularly well suited to generating antibody responses against proteins related to cardiovascular disease, neurodegenerative disease, and other age-related pathologies. The advantage of peptide-based vaccines is that they can generate antibody responses against only a small portion of the target protein, or against abnormal versions of proteins, such as misfolded proteins or abnormal proteins that are the product of splice variants, without inducing off-target antibodies.

[0007] Another application of peptide-based vaccines is to induce neutralizing antibodies against infectious organisms, such as HIV, malaria, SARS, and other coronaviruses, where a highly focused antibody response against conserved neutralizing sites may be required to prevent infection.

[0008] In addition, peptide-based vaccines may be administered alone or in combination with immunosuppressants to induce tolerance for the treatment of allergy and autoimmunity.

[0009] Although numerous peptide-based vaccine technologies have been developed, a major challenge is ensuring consistent production of uniform formulations, given the tremendous physiochemical heterogeneity of peptide antigens. Improved formulation and delivery approaches are therefore needed to ensure consistent formulations with any peptide antigen.

[0010] Another challenge is focusing antibody responses against minimal immunogens corresponding to fragments of full-length protein antigens. Such an approach may be required for the development of successful prophylactic vaccination strategies against certain highly variable viruses, thus requiring focused antibody induction against conserved sites. Minimal immunogen vaccines may also be beneficial for the treatment of certain neurodegenerative diseases, cardiovascular diseases, and cancers, where focused antibody responses may be required to balance safety and efficacy. However, a current challenge is that most vaccine platforms, such as whole organism, expression systems (e.g., DNA and viral), and protein subunit vaccines, generate broad antibody responses and are not well suited to focusing antibodies against minimal immunogens, while the most commonly used minimal immunogen vaccine platforms, i.e., KLH and virus-like particles, rely on the conjugation of synthetic peptides to recombinant carriers, often resulting in formulation variability and increased costs due to the use of recombinant manufacturing processes. Therefore, an alternative, preferably fully synthetic, system for presenting minimal immunogens is needed.

[0011] Finally, there has been growing interest in the use of peptide-based vaccines to induce tolerance for treating allergies and autoimmunity, but currently used PLGA particles and liposomes rely on empirical formulation processes that are not well suited to producing particle formulations with the multiple peptide antigens and immunomodulators that may be required. Furthermore, the optimal combination of peptide antigens and immunomodulators is unknown. Therefore, improved methods for delivering multiple immunomodulators and peptide antigens in particles to induce tolerance for the treatment of allergies and autoimmunity are needed. Improved compositions and methods for producing vaccines that address the above-mentioned challenges are discussed herein. Summary of the Invention [Means for solving the problem]

[0012] overview The present disclosure provides a method for preparing a compound comprising an amphiphile having the formula S-[B]-[U]-H, at least one peptide-antigen conjugate having a formula selected from [S]-[E1]-A-[E2]-[U]-H and H-[U]-[E1]-A-[E2]-[S]; With respect to vaccines containing During the ceremony, S, independently for each occurrence, is a solubilizing block; B is a spacer, H, independently for each occurrence, is a hydrophobic block; one or more drug molecules (D) are optionally attached to each H, either directly or via a suitable linker X1; A is, independently for each occurrence, a peptide antigen; E1, independently for each occurrence, is an N-terminal extension, E2 is, independently for each occurrence, a C-terminal extension, U is, independently for each occurrence, a linker; [ ] 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; The amphiphile and / or at least one peptide antigen conjugate comprises a dendron amplifier.

[0013] The present disclosure provides a method for producing an amphiphile having the formula S-[B]-[U]-H, and at least one peptide antigen conjugate having a formula selected from [S]-[E1]-A-[E2]-[U]-H and H-[U]-[E1]-A-[E2]-[S]. It also relates to a vaccine for inducing tolerance, comprising During the ceremony, S, independently for each occurrence, is a solubilizing block; B is a spacer, H, independently for each occurrence, is a hydrophobic block; one or more drug molecules (D) are optionally attached to each H, either directly or via a suitable linker X1; A is, independently for each occurrence, a peptide antigen; E1, independently for each occurrence, is an N-terminal extension, E2 is, independently for each occurrence, a C-terminal extension, U is, independently for each occurrence, a linker; [ ] 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; the amphiphile and / or at least one peptide-antigen conjugate comprises a dendron amplifier; The at least one peptide antigen is selected from an autoantigen, an alloantigen and an allergen.

[0014] The present disclosure also relates to a vaccine comprising an amphiphile having the formula S-[B]-[U]-H and at least one peptide antigen conjugate having a formula selected from [S]-[E1]-A-[E2]-[U]-H and H-[U]-[E1]-A-[E2]-[S], During the ceremony, S, independently for each occurrence, is a solubilizing block; B is a spacer, H, independently for each occurrence, is a hydrophobic block; one or more drug molecules (D) are optionally attached to each H, either directly or via a suitable linker X1; A is, independently for each occurrence, a peptide antigen; E1, independently for each occurrence, is an N-terminal extension, E2 is, independently for each occurrence, a C-terminal extension, U is, independently for each occurrence, a linker; [ ] 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; the amphiphile and / or at least one peptide-antigen conjugate comprises a dendron amplifier; At least one A comprises a sequence in which one or more cysteine ​​residues are replaced with alpha amino-butyric acid and / or one or more methionine residues are replaced with norleucine.

[0015] The present disclosure also relates to vaccines 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 amino-butyric acid and / or one or more methionine residues are replaced with norleucine.

[0016] The present disclosure also relates to a vaccine comprising at least one peptide-antigen conjugate having a formula selected from [S]-[E1]-A-[E2]-[U]-H and H-[U]-[E1]-A-[E2]-[S], During the ceremony, S, independently for each occurrence, is a solubilizing block; H, independently for each occurrence, is a hydrophobic block; one or more drug molecules (D) are optionally attached to each H, either directly or via a suitable linker X1; A is, independently for each occurrence, a peptide antigen; E1, independently for each occurrence, is an N-terminal extension, E2 is, independently for each occurrence, a C-terminal extension, U is, independently for each occurrence, a linker; where: [ ] indicates that the group is optional; - indicates that two adjacent groups are covalently linked to each other directly or indirectly via a suitable linker X; the at least one peptide antigen is selected from an autoantigen, an alloantigen and an allergen, and the at least one drug molecule D is present and is selected from an inhibitor of mTOR; or The at least one peptide antigen is a tumor antigen or an infectious disease antigen, at least one drug molecule D is present and is selected from agonists of TLR-3, TLR-7, TLR-8, TLR-7 / 8 and STING, and a second drug molecule (D2) is present and is selected from inhibitors of mTORC1.

[0017] In certain embodiments of the above vaccines, at least one A comprises a sequence in which one or more cysteine ​​residues are replaced with alpha amino-butyric acid and / or one or more methionine residues are replaced with norleucine.

[0018] The present disclosure also relates to a vaccine 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 amino-butyric acid and / or one or more methionine residues are replaced with norleucine.

[0019] The present disclosure also relates to a vaccine comprising at least one peptide-antigen conjugate having a formula selected from [S]-[E1]-A-[E2]-[U]-H and H-[U]-[E1]-A-[E2]-[S], During the ceremony, S, independently for each occurrence, is a solubilizing block; H, independently for each occurrence, is a hydrophobic block; one or more drug molecules (D) are optionally attached to each H, either directly or via a suitable linker X1; A is, independently for each occurrence, a peptide antigen; E1, independently for each occurrence, is an N-terminal extension, E2 is, independently for each occurrence, a C-terminal extension, U is, independently for 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, and at least one D is present and selected from an agonist of TLR-7 / 8, and the vaccine further comprises a second drug molecule (D2) selected from an inhibitor of mTOR; (iii) at least one A is a glycopeptide; or (iv) at least one A is selected from an autoantigen, an allergen, and an alloantigen, and at least one D is present and selected from an ATP-competitive mTOR inhibitor; Either [ ] 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.

[0020] The present disclosure also relates to a vaccine comprising an expression system comprising DNA or RNA encoding at least one peptide antigen (A), further comprising at least one drug molecule (D) selected from a Treg-promoting immunomodulator.

[0021] The present disclosure also relates to a peptide antigen conjugate having a formula selected from S-[E1]-A-[E2]-[U]-H-[D] and [D]-H-[U]-[E1]-A-[E2]-S, or a peptide antigen fragment having a formula selected from S-[E1]-A-[E2]-[U1] and [U1]-[E1]-A-[E2]-S, During the ceremony, S is a solubilizing block, H is a hydrophobic block, one or more drug molecules (D) are optionally attached to each H, either 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, [ ] indicates that the group is optional; - indicates that two adjacent groups are covalently linked to each other directly or indirectly via a suitable linker X; S comprises one or more amino acids.

[0022] The present disclosure also relates to a method of inducing an immune response in a subject in need thereof, comprising administering to the subject at least one dose of a first vaccine (V1), followed by at least one dose of a second vaccine (V2), where V1 is a vaccine disclosed herein and V2 is a viral vaccine. [Brief explanation of the drawings]

[0023] [Figure 1] Figure 1 shows the effect of varying (i) net charge, (ii) spacer composition (PEG vs. peptide), (iii) spacer length, and (iv) hydrophobic block length on the hydrodynamic behavior of linear amphiphiles of formula SBUH. The data show that increasing net charge promotes nanoparticle micellization, that amphiphiles with ethylene oxide (PEG)-containing spacers, B, are less likely to form aggregates and / or supramolecular associations compared to amphiphiles with amino acid-containing spacers, and that the net charge required to induce micellization in linear amphiphiles increases with increasing hydrophobic block (H) length and / or hydrophobic surface area.

[0024] [Figure 2] FIG. 2 shows a schematic diagram of amphiphilic carriers of formula SB-[U]-HD with either (A) linear, (B) dendron, or (C) brush structures.

[0025] [Figure 3] Figure 3 shows the effect of varying solubilizing block functional group (i) composition, (ii) number, and (iii) net charge on the hydrodynamic behavior of amphiphiles of formula SBUHD with either linear, dendron, or brush structures. The groups NH, COOH, OH, and Man refer to primary amine, carboxylic acid, hydroxyl, and mannose groups, respectively. Particle size was assessed by dynamic light scattering, and the number-average particle size in diameter (d, nm) is reported.

[0026] [Figure 4] FIG. 4 shows a schematic diagram illustrating a vaccine composition based on a linear amphiphilic carrier of formula SB-[U]-HD mixed with a peptide antigen conjugate of formula A-[E2]-[U]-HD.

[0027] [Figure 5] Figure 5 shows the effect of various solubilizing blocking functional groups (i) composition, (ii) number, and (iii) net charge on the hydrodynamic behavior of a vaccine containing a peptide antigen conjugate of formula AUHD mixed with an amphiphile of formula SBUHD with either a linear, dendron, or brush structure, where the molar ratio of peptide antigen conjugate to amphiphile is 1:1. The peptide antigen conjugate is compound 198. Note that amphiphiles with linear structures require a net charge greater than or equal to +4 or less than or equal to -4 to induce stable nanoparticle micelles, whereas amphiphiles with dendron structures, which have a net neutral charge, formed nanoparticle micelles with more uniform size and consistency than amphiphiles with brush structures. Particle size was assessed by dynamic light scattering, and the number-average particle size in diameter (d, nm) is reported.

[0028] [Figure 6]Figure 6 shows the T cell responses induced in mice (n = 3 / group) after vaccination with different cancer vaccine compositions or controls. Mice were vaccinated with either a vehicle control (column 1), nanoparticles containing a peptide antigen conjugate of the formula S-E1-A-E2-UHD (i.e., compound 249, column 2), or mosaic nanoparticles containing peptide antigen conjugates of the formula AUHD and AU-E2-HD (compounds 202 and 225, respectively) and an amphiphile of the formula SBUHD with either a net positive charge and a linear structure (compound 245, column 3), a net negative charge and a linear structure (compound 246, column 4), or a net neutral charge and a dendron structure (compound 162, column 5). (A) CD8 and (B) CD4 T cell responses were determined from whole blood using intracellular cytokine staining (ICS) on day 13 post-immunization. Data are reported as geometric means with 95% confidence intervals. Note that peptide-antigen conjugates Compounds 249 and 202 contain the neoantigen Adpgk, derived from the melanoma cell line MC-38, and peptide-antigen conjugate Compound 225 contains a universal CD4 T cell epitope called PADRE.

[0029] [Figure 7A] Figure 7A shows the study design and T cell responses in mice (n=5 / group) after immunization with different cancer vaccine compositions containing neoantigens (Reps1, Adpgk, Cpne1, and Irgq) used alone or in combination with vaccinia virus as an adjuvant source. (A) Treatment groups are listed in a table summarizing the compositions used for prime (first immunization on day 0) and boost (second immunization on day 14) immunizations. Groups 1-11 were treated with a vaccine containing nanoparticles further comprising a peptide-antigen conjugate, given alone or in combination with vaccinia virus ("virus"). Except for group 2, the vaccinia virus did not encode a neoantigen.

[0030] [Figure 7BC]Figures 7B-C show antigen-specific CD8 T cell responses determined from whole blood using intracellular cytokine staining (ICS) on (B) day 7 and (C) day 28 after immunization. Data are reported as geometric means with 95% confidence intervals. Note: Group 11, treated with mosaic nanoparticles containing an amphiphile with a neutral charge and dendron structure of formula SBUH and a peptide-antigen conjugate of formula AUH, induced the largest CD8 T cell response after a single immunization.

[0031] [Figure 8] Figure 8 shows a schematic diagram of mosaic nanoparticles formed by mixing an amphiphilic carrier of formula SB-[U]-HD, which has a brush structure, with a peptide antigen conjugate of formula SA-E2-[U]-HD, which has either short or long extension (E2) sequences designed to control the degree to which the antigen (A) is solvent exposed.

[0032] [Figure 9] Figure 9 shows the effect of varying peptide-antigen conjugate composition and amphiphile carrier net charge, spacer length, and structure on the hydrodynamic behavior of mosaic nanoparticle-based vaccines for inducing antibody responses. Each data point on the graph represents the particle size determined for a single mosaic nanoparticle composition containing an amphiphile of formula SBUHD and a peptide-antigen conjugate in a 1:1 molar ratio. Mosaic nanoparticles were formulated by combining the amphiphile and peptide-antigen conjugate in a 1:1 molar ratio in DMSO and then diluting the DMSO solution to a final concentration of 0.05 mM with PBS buffer, pH 7.4. Particle size was assessed by dynamic light scattering, and the number-average particle size in diameter (d, nm) is reported. Note that amphiphiles with linear structures yielded nanoparticle micelles of uniform size, while mosaic nanoparticles containing amphiphiles with brush structures showed a higher degree of particle size variability.

[0033] [Figure 10]Figure 10 shows the hydrodynamic behavior of different compositions of vaccines against SARS-CoV-2. Peptide antigen conjugates with the formula HDU-E1-A (compounds 207, 212, and 216) or peptide antigen conjugates with the formula HD-U-E1-AS containing a solubilizing block (compounds 208, 213, and 217) were mixed with a peptide antigen conjugate with the formula AUHD containing a universal CD4 T cell epitope (compound 225) and an amphiphile with the formula SBUHD at a 1:1 molar ratio of total peptide antigen conjugate to total amphiphile. Mosaic nanoparticles were formulated by combining the amphiphile and peptide antigen conjugate in a 1:1 molar ratio in DMSO and then diluting the DMSO solution to a final concentration of 0.05 mM with PBS buffer, pH 7.4. Particle size was assessed by dynamic light scattering; the number-average particle size in diameter (d, nm) is reported. It should be noted that amphiphiles with linear or dendron structures resulted in nanoparticle micelles of uniform size, while there was a higher degree of particle size variability for the mosaic nanoparticles containing amphiphiles with brush structures.

[0034] [Figure 11] Figure 11 shows anti-SARS-CoV-2 antibody titers induced with different vaccine compositions. C57BL / 6 (n=3 / group) mice were immunized with the indicated vaccine compositions on days 0 and 14, and antibody titers against three unique SARS-CoV-2 epitopes were assessed from whole blood by sandwich ELISA on day 28. The midpoint antibody titers elicited against each of the epitopes are shown.

[0035] [Figure 12] Figure 12 shows anti-SARS-CoV-2 antibody titers induced with different vaccine compositions. C57BL / 6 (n=3 / group) mice were immunized with the indicated vaccine compositions on days 0 and 14, and antibody titers against single SARS-CoV-2 epitopes were assessed from whole blood by sandwich ELISA on day 28. Midpoint antibody titers elicited against the epitopes are shown.

[0036] [Figure 13] FIG. 13 shows the chemical structures of (a) methionine (M or Met) and the amino acid norleucine (n or nLeu) used in place of methionine, and (b) cysteine ​​alpha-aminobutyric acid (B or aBut) used in place of cysteine.

[0037] [Figure 14-1]Figures 14A-E show that peptide-antigen conjugates containing a peptide antigen further comprising a T cell epitope in which a naturally occurring (or "native") methionine residue is replaced with norleucine elicit de novo T cell responses that recognize both the native antigen and the non-native antigen (i.e., the antigen in which the methionine is replaced with norleucine). (a) Sequences of the tumor autoantigen (native Trp1) and norleucine-substituted Trp1 (norTrp1) contained within compounds 257 and 258. (b) C57BL / 6 mice (n=10-15 per group) were subcutaneously (SC) vaccinated on days 0 and 14 (with 8 nmol of 257 or 258), bled on days 7 and 21, challenged subcutaneously with 105 cells of B16-F10 on day 28, and treated with anti-PD-L1 on days 35 and 42. (c) CD8 T cell responses measured by intracellular cytokine staining for IFN-γ after 6 hours of peptide stimulation in vitro on day 21. Each sample was split and stimulated with either 2.5 μg / mL of native Trp1 min or norTrp1 min. Histograms show concatenated samples of all acquisition events for each condition (left) and individual samples quantified with lines connecting paired samples (right). P values ​​are from a one-way ANOVA (Kruskal-Wallis with Dunn's correction) comparing responses to Trp1 min between groups. (d) CD8 T cell responses measured by tetramer staining with TAPDNLGYM:H-2Db-PE on day 21. Histograms show concatenated samples of all acquisition events for each vaccine group (left), and bar graphs show quantification of each individual sample (right). P values ​​are from a one-way ANOVA (Kruskal-Wallis with Dunn's correction) comparing responses across all groups. Data are mean ± standard deviation. (e) Tumor growth curves (left) and survival rates (right) after tumor challenge. P values ​​are for survival curves and are from the log-rank test with correction for multiple comparisons. [Figure 14-2] Same as above.

[0038] [Figure 15-1]Figures 15A-E show that peptide-antigen conjugates containing a peptide antigen further comprising a T cell epitope in which all three naturally occurring (or "native") methionine residues are replaced with norleucine elicit de novo T cell responses that recognize both the native antigen and the non-native antigen (i.e., an antigen in which each methionine is replaced with norleucine). (a) Sequences of tumor neoantigens (native Adpgk) and norleucine-substituted Adpgk (norAdpgk) contained within compounds 259 and 260. (b) C57BL / 6 mice (n = 10-15 per group) were subcutaneously (SC) vaccinated on days 0 and 14 (with 8 nmol of 295 or 260). (b) C57BL / 6 mice (n = 10–15 per group) were vaccinated subcutaneously (SC) on days 0 and 14, bled on days 7 and 21, challenged subcutaneously with 10 cells of B16-BFP-Adpgk on day 28, and treated with anti-PD-L1 on days 35 and 42. (c) CD8 T cell responses measured by intracellular cytokine staining for IFN-γ after 6 hours of in vitro peptide stimulation on day 21. Each sample was split and stimulated with either 2.5 μg / mL of native Adpgk min or norAdpgk min. Histograms show concatenated samples of all acquisition events for each condition (left) and individual samples quantified with lines connecting paired samples (right). P values ​​are from a one-way ANOVA (Kruskal-Wallis with Dunn's correction) comparing responses to Adpgk min between groups. (d) CD8 T cell responses measured by tetramer staining with ASMTNMELM:H-2Db-PE on day 21. Histograms show combined samples of all acquisition events for each vaccine group (left), and bar graphs show quantification of each individual sample (right). P values ​​are from one-way ANOVA (Kruskal-Wallis with Dunn's correction) comparing responses across all groups. Data are means ± standard deviations. (e) Tumor growth curves (left) and survival rates (right) after tumor challenge. P values ​​for survival curves are from the log-rank test with correction for multiple comparisons. [Figure 15-2] Same as above.

[0039] [Figure 16-1] Figures 16A-F show that peptide-antigen conjugates containing a peptide antigen further comprising a T cell epitope in which all three naturally occurring (or "native") methionine residues are replaced with norleucine, but not norvaline or leucine, elicit de novo T cell responses that recognize both the native antigen and the non-native antigen (i.e., the antigen in which each methionine is replaced with norleucine). (a) Chemical structures of methionine, norleucine, norvaline, and leucine. (b) Sequences of tumor neoantigen (native Adpgk), norleucine-substituted Adpgk (norAdpgk or norleucine-Adpgk), norvaline-substituted Adpgk (norvaline-Adpgk), and leucine-substituted Adpgk (leucine-Adpgk), contained within compounds 259, 260, 261, and 262, respectively. (c) Reverse-phase HPLC traces of compounds 259, 260, 261, and 262; the y-axis shows absorbance at 320 nm in arbitrary absorbance units (mAU). (d) C57BL / 6 mice (n = 5 per group) were subcutaneously (SC) vaccinated with 8 nmol of either 259, 260, 261, or 261 on days 0 and 14 and bled on days 7 and 21. (e) CD8 T cell responses measured after 6 hours of in vitro peptide stimulation on day 7 by tetramer staining with ASMTNMELM:H-2Db-PE (left) or intracellular cytokine staining for IFN-γ (right). Each sample was stimulated with 2.5 μg / mL of Adpgk min. P values ​​are from one-way ANOVA (Kruskal-Wallis with Dunn's correction). Data are means ± standard error. (f) CD8 T cell responses measured as in e on day 21.

[0040] [Figure 16-2] Same as above.

[0041] [Figure 17]Figures 17A-D show that peptide-antigen conjugates containing a peptide antigen further comprising a T cell epitope in which a naturally occurring (or "native") cysteine ​​residue is replaced with alpha-aminobutyric acid (B or aBut) elicit de novo T cell responses that recognize both the native antigen and the non-native antigen (i.e., the antigen in which the cysteine ​​is replaced with aBut). (a) Sequences of the viral antigen (native gp33) and aBut-substituted gp33 (aBut gp33) contained within compounds 263 and 264. (b) C57BL / 6 mice (n=9 per group) were subcutaneously (SC) vaccinated with 8 nmol of compound 263 or 264 on days 0 and 14 and bled on days 7 and 21. (c) CD8 T cell responses measured by intracellular cytokine staining for IFN-γ after 6 hours of peptide stimulation in vitro on day 21. Each sample was stimulated with 2.5 μg / mL of gp33 min. P values ​​are from one-way ANOVA (Kruskal-Wallis with Dunn's correction) comparing responses to gp33 min between groups. (d) Splenocytes were harvested (n = 4 mice per group; each symbol represents a different mouse), stimulated with the indicated concentrations of gp33 min for 6 hours, and assessed for IFN-γ by intracellular cytokine staining. The y-axis is the % IFN-γ response at a given concentration as a percent of the response at 10,000 nM. The line is a nonlinear sigmoidal regression (left). Interpolated EC50 values ​​are graphed for each mouse (right). P values ​​are from Mann-Whitney.

[0042] [Figure 18]Figures 18A-D show that peptide-antigen conjugates containing a peptide antigen further comprising a T cell epitope in which a naturally occurring (or "native") cysteine ​​residue is replaced with alpha-aminobutyric acid (B or aBut) elicit de novo T cell responses that recognize both the native antigen and the non-native antigen (i.e., the antigen in which the cysteine ​​is replaced with aBut). (a) Sequences of tumor neoantigens (native m27) and aBut-substituted m27 (aBut m27) contained within compounds 266 and 267. (b) C57BL / 6 mice (n=10 per group) were subcutaneously (SC) vaccinated with 8 nmol of compound 266 or 267 on days 0 and 14 and bled on days 7 and 21. (c) CD8 T cell responses measured by intracellular cytokine staining for IFN-γ after 6 hours of peptide stimulation in vitro on day 21. Each sample was stimulated with 2.5 μg / mL of M27 min or aBut M27 min. P values ​​are from one-way ANOVA (Kruskal-Wallis with Dunn's correction) comparing the response to M27 min between groups. (d) Memory phenotype of IFN-γ+ cells after stimulation with M27 min on day 21. Data are means ± standard deviation.

[0043] [Figure 19]Figures 19A-E show that peptide-antigen conjugates containing peptide antigens that further contain T cell epitopes in which the naturally occurring (or "native") cysteine ​​residue is replaced with serine (S or Ser) elicit significantly smaller T cell responses than those containing the native sequence. (a) Chemical structures of cysteine ​​and serine. (b) Sequences of viral antigens (native gp33) and ser-substituted gp33 (ser gp33) contained within compounds 263 and 265. (c) C57BL / 6 mice (n=5 per group) were subcutaneously (SC) vaccinated with 8 nmol of compound 263 or 265 on days 0 and 14 and bled on days 7 and 21. (d-e) CD8 T cell responses measured by intracellular cytokine staining for IFN-γ after 6 hours of peptide stimulation in vitro on days 7 (d) and 21 (e). Each sample was split and stimulated with either 2.5 μg / mL of gp33 min or ser gp33 min. Lines connect paired samples (right). P values ​​are from a one-way ANOVA (Kruskal-Wallis with Dunn's correction) comparing the response to gp33 min between groups. Note: The naive group was stimulated only with gp33 min.

[0044] [Figure 20] Figures 20A-C show that peptide-antigen conjugates containing a peptide antigen in which all three naturally occurring (or "native") cysteine ​​residues are replaced with aBut elicit de novo T cell responses that recognize both the native antigen and the non-native antigen (i.e., the antigen in which each cysteine ​​is replaced with aBut). (a) Sequences of viral antigens (native E7) and aBut-substituted E7 (aBut E7) contained within compounds 268 and 269. (b) C57BL / 6 mice (n=5 per group) were subcutaneously (SC) vaccinated with 8 nmol of compound 268 or 269 on days 0 and 14 and bled on day 21. (c) CD8 T cell responses on day 21 are shown.

[0045] [Figure 21-1]Figures 21A-21B show the pH response of exemplary amphiphiles with S blocks containing carboxylic acid solubilizing groups. (a) Amphiphile structures: Compounds 160, 270, and 271 each contain a PEG-based dendron amplifier with R25 selected from beta-alanine, glycine, and -OH. Compound 272 contains a peptide-based dendron amplifier with R26 selected from glutaric acid. Compounds 272 and 273 both contain peptide-based dendron amplifiers with R26 selected from succinic acid, except for compound 273, where the spacer B is absent. U contains a triazole, and the H block is Ahx-2B3W2. (b) Absorbance (turbidity) at 490 nm was evaluated for a range of different amphiphiles at 0.1 mM in PBS buffer at the indicated pH. In this study, absorbance values ​​greater than approximately 0.04 indicate aggregation. [Figure 21-2] Same as above.

[0046] [Figure 22-1] 22A-22B show the effect of peptide-antigen conjugate net charge, the presence of amphiphile, and amphiphile spacer (B) length on the zeta potential, hemolysis, and particle size stability of different vaccine compositions. The results show that (a) decreasing the net charge and the presence of a neutral amphiphile reduces RBC lysis, and (b) increasing the net charge leads to improved particle size stability. [Figure 22-2] Same as above.

[0047] [Figure 23] Figure 23 shows the effect of peptide antigen conjugate net charge on particle size stability of vaccine compositions with various amphiphile compositions. The results show that increasing peptide antigen conjugate net charge leads to improved particle size stability that is independent of amphiphile composition.

[0048] [Figure 24]Figure 24 shows the effect of peptide antigen conjugate net charge and the presence of amphiphile on particle size stability of vaccine compositions subjected to multiple freeze-thaw cycles. The results show that increasing peptide antigen conjugate net charge leads to improved particle size stability, and that formulations containing amphiphiles with PEG24 (or PEG36, data not shown) spacers have greater particle size stability compared to those with PEG48 spacers, which show increased aggregation tendency (indicated in this study by an absorbance ("turbidity") greater than 0.05).

[0049] [Figure 25] FIG. 25 shows the effect of different inhibitors and / or immunostimulatory agents on the percentage of T cells expressing either Tbet or Foxp3.

[0050] [Figure 26] Figures 26A-B show the effect of Torin-1, an ATP-competitive mTOR inhibitor, on the magnitude of Tregs induced by tolerance vaccines with different immunostimulants. (a) The results show that Torin-1 (or "Torin") in combination with TLR-7 / 8a, 2BXy, resulted in the largest magnitude of Treg responses, and (b) these responses were two-fold greater than those induced by Torin-1 alone and eight-fold greater than vaccines without Torin-1. Note: The concentration of peptide antigen conjugate was 500 nM, and the molar ratio of total peptide antigen conjugate to Treg-promoting immunomodulator was 1:1 for 500 nM inhibitor, 1:0.25 for 125 nM inhibitor, and 1:0.1 for 50 nM inhibitor. The results show that a vaccine for inducing tolerance comprising the exemplary mTORC1 / 2 inhibitor Torin1 results in a higher percentage of antigen-specific CD4 T cells expressing FOXP3 compared to a vaccine comprising the exemplary RORγt inhibitor (SR1555), the exemplary mTORC1 inhibitor rapamycin, and the exemplary AHR agonist ITE.

[0051] [Figure 27-1]Figures 27A-27B show the effect of E2 composition on the antibody response generated against a peptide antigen (A) that also contains a minimal immunogen. (a) shows the vaccination and sampling schedule, and (b) shows the magnitude of the antibody response (total IgG) against the SARS-CoV2 spike protein assessed from the serum of immunized mice 28 days after the first vaccination. [Figure 27-2] Same as above.

[0052] [Figure 28-1] Figures 28A-28B show the effects of E1 and / or E2 on T cell activation in vitro. (a, b) In vitro antigen presentation was assessed by incubating different peptide antigen fragments of the formula [S]-[E1]-A-[E2]-[U1] or [U1]-[E1]-A-[E2]-[S] (where A is RAHYNIVTF or AQLANDVVL) for (a) and (b), respectively, over a range of concentrations (0.001-10,000 nmol) with CD8 T cells and measuring IFN-γ responses by intracellular cytokine staining. The EC50 for each peptide antigen fragment was determined by curve fitting to the data, expressed as % maximal response versus concentration, and calculating the concentration at half-maximal response. [Figure 28-2] Same as above.

[0053] [Figure 29-1] Figures 29A-29B show how the vaccine regimen (allogeneic or heterologous prime-boost) and route of boost affect T cell responses. (a) shows the vaccination and sampling schedule. Note: Peptide-antigen conjugates were administered at 32 nmol / mouse in 100 μl PBS, and ChAdOx was administered at 1e8 infectious units (IU) in 100 μl PBS. (b) shows the magnitude of antigen-specific T cells (% tetramer+ of total CD8+ T cells) induced by different vaccine regimens as assessed by flow cytometry on days 7 and 28. [Figure 29-2] Same as above.

[0054] [Figure 30-1] Figures 30A-30B show how route and vaccine regimen (allogeneic or heterologous prime-boost) affect T cell responses. (a) shows the vaccination and sampling schedule. Note: Peptide-antigen conjugates were administered IV at 32 nmol / mouse in 100 μL PBS or IM at 8 nmol / mouse in 100 μL PBS, and ChAdOx was administered at 1e8 infectious units (IU) in 100 μL PBS. (b) shows the magnitude of antigen-specific T cells (% tetramer+ of total CD8+ T cells) induced by different vaccine regimens assessed by flow cytometry on days 7 and 28. [Figure 30-2] Same as above.

[0055] [Figure 31-1] Figures 31A-31B show how peptide-antigen conjugate net charge and / or the presence of amphiphiles affect T cell responses. (a) Vaccination and sampling schedule. Note: Peptide-antigen conjugates were administered at 32 nmol / mouse in 100 μl PBS, and ChAdOx was administered at 1e8 infectious units (IU) in 100 μl PBS. (b) shows the magnitude of antigen-specific T cells (% tetramer+ of total CD8+ T cells) induced by different vaccine regimens as assessed by flow cytometry on days 7 and 28. [Figure 31-2] Same as above.

[0056] [Figure 32-1]Figures 32A-32B show that ChAdOx was used as a biological adjuvant. (a) Vaccination and sampling schedule. Note: Peptide-antigen conjugate was administered at 32 nmol / mouse in 100 μl PBS for prime, but 32 nmol / mouse in 100 μL PBS for boost, and ChAdOx was administered at 1e8 infectious units (IU) in 100 μl PBS. (b) shows the magnitude of antigen-specific T cells (% tetramer+ of total CD8+ T cells) induced by different vaccine regimens assessed by flow cytometry on days 7 and 28. [Figure 32-2] Same as above. DETAILED DESCRIPTION OF THE INVENTION

[0057] 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 more clarity regarding the compounds, compositions, methods, and their use. The terminology in this disclosure is understood to be useful for the purpose of providing a better description of certain embodiments, and should not be considered limiting.

[0058] About: In the context of this disclosure, "about" when referring to a measurable value, such as an amount, duration, and the like, is intended to encompass variations of ±20%, ±10%, ±5%, ±1%, or ±0.1% from the specified value, as such variations are appropriate for carrying out 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.

[0059] Administration: Providing or giving to a subject an agent, e.g., an immunogenic composition comprising an amphiphilic block copolymer and a drug described herein, by any effective route. Exemplary routes of administration include, but are not limited to, oral, injection (e.g., subcutaneous, intramuscular, intradermal, intraperitoneal, and intravenous), transdermal, topical, intranasal, vaginal, and inhalation routes.

[0060] "Administration of" a compound and "administering" should be understood to mean providing a compound, a prodrug of a compound, or a pharmaceutical composition described herein. The compound or composition may be administered to the subject by another person or may be self-administered by the subject.

[0061] Antigen: Any molecule that contains an epitope that binds to a T cell or B cell receptor and can stimulate an immune response, particularly a B cell response and / or a T cell response, in a subject. Epitopes can include peptides, glycopeptides, lipids, or any suitable molecule that contains an epitope that can interact with a component of a specific B cell or T cell protein. Such an interaction can generate a response by an immune cell. "Epitope" refers to the region of a peptide antigen with which a B and / or T cell receptor, i.e., a B cell receptor and a T cell receptor, interacts. Antigens used in embodiments of the present disclosure can be selected from pathogens, cancerous cells, autoantigens, alloantigens, or allergens. Many such antigens can be used in accordance with the inventive embodiments of the present disclosure, and many such antigens are discussed in more detail throughout this specification.

[0062] 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.

[0063] Amphiphilic: The term "amphiphilic" is used herein to mean a substance that contains both hydrophilic or polar groups and hydrophobic groups.

[0064] 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 called helper T cells or CD4 T cells.

[0065] 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 called cytotoxic T cells (CTLs), killer T cells, or CD8 T cells.

[0066] Charge: A physical property of a substance that affects its interactions with other atoms and molecules (including solutes and solvents). Charged substances are subject to 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 of the same charge repel each other, while two charged molecules of different charges attract each other. Charge is often described as a positive or negative integer. A molecule's charge can be easily estimated based on the molecule's Lewis structure and accepted methods known to those skilled in the art. Charge can also result from inductive effects; for example, atoms bonding together with different electron affinities can result in polar covalent bonds that result in partially negatively charged atoms and partially positively charged atoms. For example, nitrogen bonding with hydrogen results in a negative partial charge on the nitrogen and a positive partial 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 adding up the charges of each atom that constitutes the molecule. Those skilled in the art are familiar with the process of estimating the charge of a molecule by adding up 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 that may be positive or negative, and may have a partial or full integer charge, 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 that 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 contain 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 comprising amphiphile and / or peptide antigen conjugates that further comprise charged groups, e.g., charged monomers, e.g., charged amino acids, can be estimated by summing the charges of each functional group within the amphiphile and / or peptide antigen conjugate.

[0067] Chemotherapeutic Agent: A type of drug molecule (D) broadly defined as any pharmaceutically active molecule useful in the treatment of cancer, including growth inhibitory or cytotoxic agents, such as alkylating agents, antimetabolites, anti-microtubule inhibitors, topoisomerase inhibitors, receptor tyrosine kinase inhibitors, angiogenesis inhibitors, and the like. Examples of chemotherapeutic agents include: alkylating agents, such as thiotepa and cyclophosphamide (CYTOXAN®); alkyl sulfonates, such as busulfan, improsulfan, and piposulfan; aziridines, such as benzodopa, carboquone, meturedopa, and uredopa; ethyleneimines and methylmelamines (altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramine); and trimethylolmelamine); nitrogen mustards, such as chlorambucil, chlornaphazine, cyclophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novembucin, fenesterine, prednimustine, trofosfamide, uracil mustard; nitrosoureas, such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, ranimustine; anti-inflammatory drugs Biologics such as aclacinomycin, actinomycin, austramycin, azaserine, bleomycin, cactinomycin, calicheamicin, carabicin, carminomycin, carzinophilin, chromomycin, dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, doxorubicin, epirubicin, esorubicin, idarubicin, marcellomycin, mitomycin, mycophenolic acid, nogalamycin, ointmentheptamin ... Ribomycin, peplomycin, potfilomycin, puromycin, queramycin, rodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin, zorubicin; antimetabolites such as methotrexate and 5-FU; folic acid analogs such as denopterin, methotrexate, pteropterin, trimetrexate; purine analogs such as fludarabine, 6-mercaptopurine, thiamiprine, thioguanine;Pyrimidine analogues, such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine; androgens, such as calsterone, dromostanolone propionate, epithiostanol, mepitiostane, testolactone; adrenocortical hormone synthesis inhibitors, such as aminoglutethimide, mitotane, trilostane; folic acid supplements, such as folinic acid acid); aceglatone; aldophosphamide glycoside; aminolevulinic acid; amsacrine; bestravcil; bisantrene; edatrexate; defofamine; demecolcine; diaziquone; eflornithine; elliptinium acetate; etoglucide; gallium nitrate, hydroxyurea; lentinan; lonidamine; mitoguazone; mitoxantrone; mopidamol; nitracrine; pentostatin; fenamet; pirarubicin; podophyllic acid; 2-ethylhydrazide; procarbazine; PSK®; razoxane; sizofiran; spirogermanium; tenuazonic acid; triaziquone; 2,2',2"-trichlorotriethylamine; urethane; vindesine; dacarbazine; mannomustine; mito Bronitol; Mitolactol; Pipobroman; Gacytosine; Arabinoside ("Ara-C"); Cyclophosphamide; Thiotepa; Members of the taxoid or taxane family, e.g., paclitaxel (TAXOL®), docetaxel (TAXOTERE®) and their analogs; Chlorambucil; Gemcitabine; 6-thioguanine; Mercaptopurine; Methotrexate; Platinum analogs, e.g., cisplatin and carboplatin; Vinblastine; Platinum; Etoposide (VP-16); Ifosfamide; Mitomycin C; Mitoxantrone; Vincristine; Vinorelbine; Navelbine; Novantrone; Teniposide; Daunomycin; Aminopterin; Xeloda; Ibandronate; CPT-11; The topoisomerase inhibitor RFS 2000; difluoromethylornithine (DMFO); retinoic acid; esperamicin; capecitabine;Receptor tyrosine kinase and / or angiogenesis inhibitors (sorafenib (NEXAVAR®), sunitinib (SUTENT®), pazopanib (VOTRIENT™), toceranib (PALLADIA™), vandetanib (ZACTIMA™), cediranib (RECENTIN®), regorafenib (BAY 73-4506), axitinib (AG013736), lestaurtinib (CEP-701), erlotinib (TARCEVA®), gefitinib (IRESSA™), BIBW 2992 (TOVOK™), lapatinib (TYKERB®), neratinib (HKI-272), and the like), and pharmaceutically acceptable salts, acids, or derivatives of any of the foregoing. Also included within this definition are antihormonal drugs that act to regulate or inhibit hormone action on tumors, such as antiestrogens (including, for example, tamoxifen, raloxifene, aromatase-inhibiting 4(5)-imidazoles, 4-hydroxytamoxifen, trioxifene, keoxifene, LY 117018, onapristone, and toremifene (FARESTON®)); and antiandrogens, such as flutamide, nilutamide, bicalutamide, leuprolide, and goserelin; and pharmaceutically acceptable salts, acids, or derivatives of any of the foregoing. Other conventional cytotoxic chemicals, such as those disclosed in Wiemann et al., 1985, in Medical Oncology (Calabresi et al., eds.), Chapter 10, McMillan Publishing, are also suitable chemotherapeutic agents.

[0068] A chemotherapeutic drug (also called a chemotherapeutic agent) is a pharmaceutically active compound and is therefore sometimes referred to generally herein as a drug, or a drug molecule, or as "D" in a formula. For clarity, the terms chemotherapeutic drug and chemotherapeutic agent are used herein to describe any synthetic or naturally occurring molecule useful in cancer treatment, although certain classes of drug molecules may alternatively be described by their mechanism of action; for example, angiogenesis inhibitors are chemotherapeutic drugs that inhibit angiogenesis. Certain immunomodulators, such as immunostimulators, may be useful in cancer treatment, but immunomodulators, including immunostimulators and immunosuppressants, are not referred to herein as chemotherapeutic drugs.

[0069] Click chemistry reaction: A bioorthogonal reaction that couples two compounds together under mild conditions in a high-yielding 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.

[0070] Copolymer: A polymer derived from two (or more) different monomers, as opposed to a homopolymer, in which only one monomer is used. Because copolymers contain at least two types of constitutional units (also structural units), copolymers can be classified based on how these units are arranged along the chain. A copolymer can be a statistical (or random) copolymer, in which the two or more monomer units are randomly distributed; a copolymer can be an alternating copolymer, in which two or more monomer units are distributed in an alternating sequence; or, for example, copolymers such as poly(amino acids) produced by solid-phase peptide synthesis (SPPS) can have a specific order of monomer units. The term "block copolymer" generally refers to a polymer composed of two or more building blocks of different constitutional monomers or comonomers (where one block contains two or more different monomers). A block copolymer may be used herein to refer to a copolymer comprising two or more homopolymer subunits, a copolymer comprising two or more copolymer subunits, or a copolymer comprising 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 via intermediate non-repeating subunits, e.g., a joining 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" or "triblock copolymers," respectively. Copolymers may be generally referred to as polymers; for example, statistical copolymers may be referred to as polymers or copolymers. Similarly, block copolymers may be generally referred to as polymers. As used herein, a copolymer refers to a polymer comprising two or more types of monomers, while a terpolymer is a copolymer having three monomer units.

[0071] Critical micelle concentration (CMC): refers to the concentration of a material above which micelles spontaneously form to establish thermodynamic equilibrium.

[0072] Drug: refers to any pharmaceutically active molecule, including but not limited to proteins, peptides, sugars, saccharides, nucleosides, inorganic compounds, lipids, nucleic acids, small synthetic chemicals, and macrocyclic molecules, 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, antineoplastic 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, cough suppressants (expectorants and mucolytics), diagnostic agents, imaging diagnostic agents. Drugs may be selected from a variety of known classes of compounds, including diuretics, dopamine agonists (anti-Parkinson's agents), free radical scavengers, growth factors, hemostatic agents, immunological agents, lipid regulating agents, muscle relaxants, proteins, e.g., 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), antiallergic agents, stimulants and appetite suppressants, steroids, sympathomimetics, thyroid medications, vaccines, vasodilators, and xanthines. Drugs may also be referred to as pharmaceutically active agents, pharmaceutically active substances, biologically active compounds, or biologically active molecules. Any drug molecule in the formulas described herein is abbreviated as "D."

[0073] Drug delivery: The method or process of administering a pharmaceutical compound to achieve a therapeutic effect in a human or animal.

[0074] 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 herein, described herein, or exemplified herein. Such a result includes, but is not limited to, an effective reduction in symptoms associated with any of the conditions referred to herein, as determined by any suitable means in the art.

[0075] Graft copolymer: A polymer having a main polymer chain (e.g., polymer A) with one or more side chains of a second polymer (e.g., polymer B). The first polymer A is linked to the second polymer B by its monomers and side chains, and polymer B is attached to individual monomers of polymer A and thus branches off from the polymer A chain. A first polymer linked to a second polymer by end groups is sometimes described as a block polymer (e.g., AB diblock) or end-graft polymer.

[0076] Hydropathy index / GRAVY value: A number representing the hydrophobic or hydrophilic properties of an amino acid or a sequence of amino acids. There are various scales that can be used to represent the relative hydrophobic and hydrophilic properties of the amino acids that make up a peptide. In this 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 (GRAVY) value, 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. A peptide sequence with a higher GRAVY value is considered to be more hydrophobic, while a peptide sequence with a lower GRAVY value is considered to be more hydrophilic.

[0077] Hydrophilicity: Refers to the tendency of a material to be freely dispersed or solubilized in an aqueous solution (sometimes called an aqueous medium). A material is considered hydrophilic if it prefers to interact with other hydrophilic materials and avoids interacting with hydrophobic materials. In some cases, hydrophilicity is used as a relative term; for example, the same molecule may or may not be described as hydrophilic, 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 polar and / or charged portion of a molecule that has good aqueous solubility.

[0078] 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 interacting with hydrophilic materials. Hydrophobicity is a relative term; the same molecule may or may not be described as hydrophobic, depending on what it is being compared to. Hydrophobic molecules are often non-polar and uncharged and have poor water solubility, e.g., insoluble in water or soluble in water only at concentrations of 1 mg / mL or less, usually 0.1 mg / mL or less, or more preferably 0.01 mg / mL or less. A hydrophobic monomer is a monomer containing a hydrophobic group that forms a water-insoluble polymer or a polymer that is insoluble in water at a certain temperature, pH, and salt concentration, such as a hydrophobic amino acid. The hydrophobic group refers to the portion of a molecule that is hydrophobic. For example, a styrene monomer is sometimes called a hydrophobic monomer because poly(styrene) is a water-insoluble polymer. Hydrophobic drugs refer to drug molecules that are insoluble in aqueous solutions at a pH of about pH 7.4 or that are soluble only at concentrations of about 1.0 mg / mL or less. Amphipathic drugs are drug molecules that tend to aggregate into supramolecular structures, such as micelles, in aqueous solutions and / or have limited solubility in aqueous solutions at a pH of about pH 7.4.

[0079] 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 (e.g., via 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, e.g., 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 may be used to stimulate an immune response, resulting in the activation of cytotoxic T cells that kill virally infected or cancerous cells. In other embodiments, an antigen may be used to induce tolerance or immunosuppression. A tolerogenic response may result from the unresponsiveness of T cells or B cells to an antigen. Suppressive immune responses can also result from the priming and / or activation of regulatory cells, such as regulatory T cells, or the transdifferentiation of effector cells into regulatory cells, which downregulate, i.e., attenuate, the immune response.

[0080] Immunogenic composition: A formulation of material containing an antigen and, optionally, an immunomodulatory substance that induces a measurable immune response to the antigen. For example, a vaccine is a type of immunogenic composition.

[0081] Immunomodulator: Refers to a class of drugs that modulate the activity of cells of the immune system, including immunostimulants and immunosuppressants.

[0082] Immunostimulant: refers to any synthetic or naturally occurring drug that promotes the pro-inflammatory and / or cytotoxic activity of immune cells. Exemplary immunostimulants include pattern recognition receptor (PRR) agonists, such as synthetic or naturally occurring agonists of Toll-like receptors (TLRs), 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 (such as certain interleukins), such as IL-2; certain chemokines or small molecules that bind to chemokine receptors; certain antibodies, antibody fragments, or synthetic peptides that activate immune cells, for example, by binding to stimulatory receptors, such as anti-CD40, or certain antibodies, antibody fragments, or synthetic peptides that activate immune cells, for example, by blocking inhibitory receptors, such as anti-CTLA4, anti-PD1, etc. Various immunostimulatory agents 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 immunostimulatory agents, or more commonly as drug molecules (abbreviated as "D" in formulas).

[0083] Immunosuppressant: refers to any synthetic or naturally occurring drug that suppresses pro-inflammatory and / or cytotoxic activity by immune cells or the humoral immune system, e.g., antibodies and complement proteins. Immunosuppressants may mediate their effects by one or more of the following mechanisms: priming suppressor cells, e.g., regulatory T cells; killing, inhibiting, or inactivating pro-inflammatory, cytotoxic, and / or B cells; transdifferentiating pro-inflammatory and / or cytotoxic T cells into suppressor cells; and / or sequestering pro-inflammatory, cytotoxic, and / or B cells and / or limiting their mobility. 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 may be more commonly referred to as drug molecules (abbreviated as "D" in the formula).

[0084] In vivo delivery: Administration of a composition, e.g., a composition comprising an amphiphilic block copolymer and a drug, 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.

[0085] Linked or Coupled: The terms "linked" and "coupled" mean attached to one another, 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 linked to another molecule by a covalent bond. 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 also be linked to another molecule by any and all combinations of such couplings. Molecules may also be linked indirectly, for example, by the use of a linker (sometimes referred to as a linker molecule). Molecules may also be indirectly linked through a moiety that is independently non-covalently bound to both molecules. As used herein in chemical formulas, the term "linker," sometimes abbreviated as "X," refers to any suitable linker molecule. Specific, preferred linkers may also be designated by other symbols, such as X, X, X, X, X, X, and U. A variety of linkers are described throughout this specification.

[0086] A "bilayer membrane" or "bilayer" is a self-assembled film of amphiphiles or supraamphiphiles in aqueous solution.

[0087] Micelle: A spherical container with a single monolayer that defines a closed compartment. Generally, amphiphilic molecules spontaneously form micellar structures in polar solvents. In contrast to bilayers, e.g., lipid bilayers, micelles are "sided" because they project a hydrophilic, polar exterior and present a hydrophobic interior.

[0088] 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 where the density (or mol%) of monomer A equals 10 mol%, will have 10 monomer units of A, and the remaining 90 monomer units (or "monomers") may be monomer B or another monomer, unless otherwise specified.

[0089] 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 that of a monomer. 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 amino acid monomer units. A monomeric unit may also be called a monomer or a monomer unit, or the like.

[0090] Net Charge: The total electrostatic charge carried by a molecule, or, where specified, a section of a molecule.

[0091] Particle: A nano- or micro-sized supramolecular structure composed of an aggregate 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 can be larger than 100 nm.

[0092] 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. There are several classes of PRRs. Non-limiting examples of PRRs include Toll-like receptors (TLRs), RIG-I-like receptors (RLRs), NOD-like receptors (NLRs), stimulator of interferon genes (STING) receptors, and C-type lectin receptors (CLRs). Agonists of such PRRs are called immunostimulants and can be used to enhance and / or regulate immune responses to antigens. For more information on pattern recognition receptors, see Wales et al., Biochem Soc Trans., 35:1501-1503, 2007.

[0093] Peptide or Polypeptide: Two or more natural or unnatural amino acid residues linked together in series by one or more amide bonds. The amino acid residues may have post-translational modifications (e.g., glycosylation, citrullination, homocitrullination, oxidation, and / or phosphorylation). Such modifications may mimic post-translational modifications that occur naturally in vivo or may be unnatural. Any one or more of the components of the amphiphile and / or peptide antigen conjugate may comprise a peptide.

[0094] Peptide modifications: Peptides may be altered or otherwise synthesized using one or more of several modifications, as shown below. In addition, 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 sequences of amino acids, 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 exclusively L-amino acids. Such peptides may contain naturally occurring and non-naturally occurring peptides, analogs, derivatives, and variants.

[0095] Peptides can be modified by any of a variety of chemical methods 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 CC1-CC2. 16It can be made into an ester (where CC refers to a carbon chain (thus CC1 refers to a single carbon and CC16 refers to 16 carbons)) or converted into an amide. 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, acetate, trifluoroacetate, formate, benzoate, toluenesulfonate, maleate, tartrate, and other organic salts, or can be modified or converted into an amide (e.g., by acetylation).

[0096] The peptides may be modified to contain substituents that contain positive or negative charges or both. The positive and / or negative charges may be affected by the pH at which the peptide is present.

[0097] Hydroxyl groups on the peptide side chains are cleaved from C1 to C6 using well-recognized methods. 16 Alkoxy or C1-C 16 They may be converted to esters, or the hydroxyl groups may be converted (e.g., sulfated or phosphorylated) to introduce a negative charge. The phenyl and phenol rings of the peptide side chains may be substituted with one or more halogen atoms, e.g., fluorine, chlorine, bromine, or iodine, or may be C1-C 16 Alkyl, C1-C 16Substitution with alkoxy, carboxylic acids and their esters, or amides of such carboxylic acids may be used. Methylene groups in peptide side chains can be extended to homologous 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 a number of 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 select and equip the structure with conformational constraints that result in enhanced stability. For further details on modifications that can be made to functional groups, see Greene et al., "Greene's Protective Groups in Organic Synthesis," 4th Edition, John Wiley & Sons, Inc., 2006.

[0098] The unexpected discovery reported herein is that cysteine ​​residues of naturally occurring peptide antigens can be replaced with alpha-aminobutyric acid or serine, and methionine residues with norleucine, to obtain non-natural peptide antigens that induce immune responses that cross-react with the naturally occurring peptide antigens. Preferred methods for preparing and using peptide antigens with non-native sequences are described throughout this specification.

[0099] Pharmaceutically acceptable vehicle: The pharmaceutically acceptable vehicle (or carrier) useful in the present disclosure includes conventional carriers, additives and diluents.Remington's Pharmaceutical Sciences by EW Martin, Mack Publishing Co., Easton, PA, 15th Edition (1975) describes compositions and formulations suitable for the pharmaceutical delivery of one or more therapeutic compositions, such as one or more therapeutic cancer vaccines and additional pharmaceutical agents.

[0100] Pharmaceutically acceptable carriers are well known in the art and include, for example, aqueous solutions, such as water or physiological buffered saline, or other solvents or vehicles, such as glycols, glycerol, oils, such as olive oil, or injectable organic esters. In a preferred embodiment, when such pharmaceutical compositions are intended for administration to humans, particularly for invasive administration routes (i.e., routes such as injection or placement that avoid transport or diffusion through epithelial barriers), the aqueous solutions are pyrogen-free or substantially pyrogen-free. Excipients can be selected, for example, to cause delayed release of the drug or to selectively target one or more cells, tissues, or organs. Pharmaceutical compositions can be in dosage unit form, such as tablets, capsules (including sprinkle capsules and gelatin capsules), granules, lipophilic substances for reconstitution, powders, solutions, syrups, suppositories, injections, or the like. The compositions can also be present in transdermal delivery systems, such as skin patches. The composition may also be in a solution suitable for topical administration, such as an ointment or cream.

[0101] Pharmaceutically acceptable carriers may contain physiologically acceptable agents that act to stabilize compounds, increase the solubility of compounds, or enhance absorption of compounds, such as compounds 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 choice of a pharmaceutically acceptable carrier containing a physiologically acceptable agent depends, for example, on the route of administration of the composition. The pharmaceutical composition preparation may be a self-emulsifying or self-microemulsifying drug delivery system. The pharmaceutical composition (preparation) may also be a liposome or other polymer matrix, which may incorporate, for example, a compound of the present invention. Liposomes, such as liposomes containing phospholipids or other lipids, are non-toxic, physiologically acceptable, and metabolizable carriers that are relatively easy to prepare and administer.

[0102] Polarity: A description of the properties of a substance. Polarity is a relative term and can describe a molecule or a portion of a molecule that has a partial charge resulting from the difference in electronegativity between atoms bonded to each other in the molecule, such as the bond between nitrogen and hydrogen. Polar molecules prefer to interact with other polar molecules and do not usually associate with non-polar molecules. In certain non-limiting cases, polar groups may contain hydroxyl groups, or amino groups, or carboxyl groups, or charged groups. In certain non-limiting cases, polar groups may also prefer to interact with polar solvents such as water. In certain non-limiting cases, the introduction of additional polar groups may increase the solubility of a portion of the molecule.

[0103] Polymer: A molecule containing repeating structural units (monomers). As explained in more detail throughout this disclosure, polymers can be used for any number of components for amphiphiles, peptide-antigen conjugates, and drug molecule conjugates, and can be natural or synthetic. Various compositions of polymers useful in the practice of the present invention are discussed in more detail elsewhere. Note: Polymer is used throughout this specification to broadly encompass molecules having as few as three or more monomers, which may also be referred to as oligomers.

[0104] Polymerization: A chemical reaction, usually performed with a catalyst, heat, or light, in which monomers combine to form a chain, branched, or crosslinked macromolecule (polymer). Chain, branched, or crosslinked macromolecules can be modified by further 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 of a monomer. The free radical adds to one side of the double bond, creating a free electron on the other side. This free electron then reacts with another monomer, and the chain self-propagates, thus allowing one monomer unit to be added to the end of the growing chain one at a time. 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, for example, during solid-phase peptide synthesis (SPPS).

[0105] Polymersomes: vesicles assembled from synthetic multiblock polymers in aqueous solution. Unlike liposomes, polymersomes do not contain lipids or phospholipids as their primary components. As a result, polymersomes are thermally, mechanically, and chemically distinct from the most stable lipid vesicles, and are particularly more durable and resilient. Polymersomes assemble during lamellar swelling, for example, by film or bulk rehydration, as described below, or by 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.

[0106] Purified: A substance or composition that is relatively free from impurities or substances that degrade or contaminate the substance or composition. The term purified is a relative term and does not require absolute purity. Substantially purified means purification from impurities. A substantially purified substance or composition is usually at least 60%, 70%, 80%, 90%, 95%, 98%, or 99% pure.

[0107] Soluble: Ability 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 assemble into multimers or other supramolecular structures through hydrophobic or other non-covalent interactions. Soluble molecules are understood to be freely dispersed in solution as single molecules. The hydrophobic block (H) described herein is insoluble or soluble only 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.

[0108] Subject and patient: These terms may be used interchangeably herein and refer to both humans and non-human animals, including birds 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).

[0109] 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 this disclosure. In addition, targeting molecules can also have a solubilizing effect and can be considered either a drug molecule (D) and / or a solubilizing (SG) group, or both.

[0110] T cell: a type of white blood cell that is part of the immune system and can participate in an immune response. T cells are composed of CD4 These include, but are not limited to, CD4 T cells and CD8 T cells. CD4 T cells present CD4 glycoprotein on their surface, and these cells are often called helper T cells. These cells often regulate immune responses, including antibody responses and cytotoxic T cell responses, although CD4 T cells (e.g., regulatory T cells) can also suppress immune responses, and CD4 T cells can act as cytotoxic T cells. CD8 T cells present CD8 glycoprotein on their surface, and are often called cytotoxic or killer T cells, but CD8 T cells can also suppress immune responses.

[0111] Treating, preventing, or ameliorating a disease: "Treating" refers to an intervention that reduces a sign or symptom or marker 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, which refers to a decrease in the number or size of tumors and / or metastases, or treating a disease may result in immune tolerance, which reduces systems related to autoimmunity. "Preventing" a disease means inhibiting the full development of the disease. A disease may also be prevented from occurring at all. A disease may also be prevented from progressing in severity or degree or type. "Ameliorating" refers to a reduction in the number or severity of signs or symptoms or markers of a disease, such as cancer.

[0112] Reducing the signs, symptoms, or markers of a disease or disease-related pathological condition refers to any observable beneficial effect of treatment, and / or any observable effect on a proximal surrogate endpoint, such as tumor volume, whether symptomatic or not. Reducing the signs or symptoms associated with tumors or viral infections can be demonstrated, for example, by delaying the onset of clinical symptoms of disease in susceptible subjects (e.g., subjects with tumors that have not yet metastasized, or subjects who may be exposed to viral infections), by reducing the severity of some or all clinical symptoms of disease, by slower disease progression (e.g., by extending the lifespan of subjects with tumors or infected with viruses), by reducing the number of disease relapses, by improving the subject's overall health or well-being, or by other parameters known in the art (e.g., parameters specific to a particular tumor or viral infection). A "prophylactic" 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 the onset of a pathological condition.

[0113] Tumor or cancer or neoplastic: An abnormal growth of cells that may be benign or malignant, but often does not necessarily produce clinical symptoms. "Neoplastic" cell growth refers to cell growth that is not responsive to physiological cues, such as growth and inhibitory factors.

[0114] 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, such as in some leukemias, neoplastic cells may not form a solid mass. In such cases, the collection of neoplastic cells may be referred to as a liquid cancer.

[0115] 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 neighboring cells, release of abnormal levels of cytokines or other secretory products, and suppressed or exacerbated inflammatory or immunological responses, and invasion of surrounding or distant tissues or organs, such as lymph nodes.

[0116] Tumors that do not exhibit substantial adverse clinical symptoms and / or grow slowly are termed "benign."

[0117] "Malignant" means causing or likely to cause significant clinical symptoms in the future. A tumor is said to be "malignant" if it produces substantial clinical symptoms by infiltrating surrounding tissues and / or metastasizing and / or producing and secreting chemical mediators that affect nearby or distant body systems.

[0118] "Metastatic disease" refers to cancer cells that have left the original tumor site and traveled to other parts of the body, for example, via the bloodstream, via the lymphatic system, or via body cavities such as the abdominal or thoracic cavity.

[0119] 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, radiological imaging, or pathological examination.

[0120] An "established" or "pre-existing" tumor is one that is present at the time treatment is initiated. Often, 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 about 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 are generally newly established, have a robust blood supply, and may have induced regulatory T cells (Tregs) and myeloid-derived suppressor cells (MDSCs).

[0121] Unit dose: A discrete amount of a pharmaceutical composition comprising a predetermined amount of an active ingredient.

[0122] Vesicle: A sac filled with fluid. In some embodiments, the vesicle is a sac containing an amphiphilic substance. In some embodiments, the sac is a nanoparticle-based vesicle, which refers to a vesicle having a size or dimension in the nanometer range. In some embodiments, the polymeric vesicle is a vesicle formed from one or more polymers.

[0123] 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 instances in which the event or circumstance occurs as well as instances in which it does not occur. For example, "optionally substituted alkyl" refers to an alkyl that may be substituted or unsubstituted. It is understood that the substituents and substitution patterns of 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 and 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, provided that a stable structure results. As used herein, the term "optionally substituted" refers to the replacement of one to six hydrogen radicals in a given structure with the radical of a 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 one to four hydrogen radicals in a given structure with the aforementioned substituents. More preferably, one to three hydrogen radicals are replaced by the aforementioned substituents. 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, an "alkyl" group refers to a C1-C6 straight chain alkyl group or a C1-C6 branched chain alkyl group. Most preferably, an "alkyl" group 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, neopentyl, 1-hexyl, 2-hexyl, 3-hexyl, 1-heptyl, 2-heptyl, 3-heptyl, 4-heptyl, 1-octyl, 2-octyl, 3-octyl, or 4-octyl, and the like. An "alkyl" group 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 to it. 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 chains 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 having substituents replacing a hydrogen on one or more carbons of the hydrocarbon backbone, including haloalkyl groups such as trifluoromethyl and 2,2,2-trifluoroethyl. The term “C x~y " or "C x ~C y " when used in conjunction with a chemical moiety such as acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy, is intended to include groups containing x to y carbons in the chain. CO alkyl denotes hydrogen, in which case the group is in a terminal position, and a bond if 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-. The term "amide" as used herein 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 with 4 to 8 atoms in the ring structure) Refers to... The terms "amine" and "amino" are art-recognized and refer to both unsubstituted and substituted amines and their salts, e.g., [ka] (In the formula, 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 with 4 to 8 atoms in the ring structure) This refers to a part that can be represented by The term "aminoalkyl," as used herein, refers to an alkyl group substituted with an amino group. The term "aralkyl," as used herein, 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 herein with the term "aromatic group." Unless otherwise specifically stated herein, aryl moieties are optionally substituted with one or more substituents, which substituents are independently 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(where t is 1 or 2), -S(O) t OR a (where t is 1 or 2), -S(O) t N(R a )2 (where t is 1 or 2), or PO3(R a )2, where each R a are independently hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, or heteroarylalkyl. Aromatic carbocycles include monocyclic aromatic groups in which each atom of the ring is carbon. Preferably, the ring is 5- to 7-membered, more preferably 6-membered. 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 the rings being aromatic, e.g., the other cyclic rings being 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 refers to a group [ka] (In the formula, R 22 and R 23 independently represent hydrogen or a hydrocarbyl group. Refers to... 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 and 8- to 12-membered bicyclic rings. Each ring in a bicyclic carbocycle may be selected from saturated, unsaturated, and aromatic rings. Carbocycles include bicyclic molecules in which one, two, 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 may be selected from saturated, unsaturated, and aromatic rings. For example, an aromatic ring, such as phenyl, may 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 that can have 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 -OCO2-. The term "carboxy," as used herein, refers to a group represented by the formula -CO2H. The term "ester" as used herein refers to the group -C(O)OR 22 R in this formula 22 refers to a hydrocarbyl group. The term "ether," as used herein, refers to a hydrocarbyl group linked to another hydrocarbyl group by an oxygen. Thus, an ether substituent of a hydrocarbyl group can be hydrocarbyl-O-. Ethers can be either 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 "hetaaralkyl" 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- to 6-membered rings, whose 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, e.g., the other cyclic rings are cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclyl. Heteroaryl groups include, for example, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyridine, pyrazine, pyridazine, and 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 systems whose ring structure contains at least one heteroatom, preferably 1 to 4 heteroatoms, more preferably 1 or 2 heteroatoms, and preferably include 3- to 10-membered rings, more preferably 3- to 7-membered rings. 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 are cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclyl. Heterocyclyl groups include, for example, piperidine, piperazine, pyrrolidine, morpholine, lactones, lactams, and the like. The term "hydrocarbyl," as used herein, refers to a group bonded by a carbon atom, having no =0 or =S substituents, typically having at least one carbon-hydrogen bond, and a primarily carbon backbone, but optionally containing heteroatoms. Thus, groups such as methyl, ethoxyethyl, 2-pyridyl, and even trifluoromethyl are considered hydrocarbyl herein, while substituents such as acetyl (having an =0 substituent on the linking carbon) and ethoxy (linked by oxygen rather than carbon) are not considered hydrocarbyl. Hydrocarbyl groups include, but are not limited to, aryl, heteroaryl, carbocyclic, heterocyclic, alkyl, alkenyl, alkynyl, and combinations thereof. The term "hydroxyalkyl," as used herein, 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. "Lower alkyl," for example, refers to an alkyl group containing 10 or fewer carbon atoms, preferably 6 or fewer carbon atoms. In certain embodiments, acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy substituents defined herein are lower acyl, lower acyloxy, lower alkyl, lower alkenyl, lower alkynyl, or lower alkoxy, respectively, whether they occur alone or in combination with other substituents, for example, hydroxyalkyl and aralkyl in this description (in which case, for example, atoms in an aryl group are not counted when counting the 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, in the ring. The term "sulfate" is art-recognized and refers to the group -OSO3H or a pharmaceutically acceptable salt thereof. The term "sulfonamide" is art-recognized and can be represented by the general formula [ka] (In the formula, R 22 and R 23 independently represent hydrogen or hydrocarbyl. It refers to a group represented by: 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 moieties having substituents replacing hydrogen on one or more carbons of the backbone. It will be understood that "substituted" or "substituted with" includes the implicit proviso that such substitution is subject to the permissible valence of the substituted atom and substituent, and that the substitution results in a stable compound, e.g., a stable compound that does not spontaneously undergo 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 the present invention, heteroatoms, such as nitrogen, can have hydrogen substituents and / or any permissible substituents of organic compounds described herein that satisfy the valence of the heteroatom. Substituents can include any of the substituents described herein, for example, halogen, hydroxyl, carbonyl (e.g., carboxyl, alkoxycarbonyl, formyl, or acyl), thiocarbonyl (e.g., 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. It will be understood by those skilled in the art 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. The term "thioester" as used herein refers to the group -C(O)SR 22 or -SC(O)R 22 R in the formula 22 refers to hydrocarbyl. The term "thioether," as used herein, is equivalent to an ether, where the oxygen has been replaced by a sulfur. The term "urea" is art-recognized and has the general formula [ka] (In the formula, R 22 and R 23 independently represent hydrogen or hydrocarbyl. It can be represented by: 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 prototypical aromatic amino acids.

[0124] Those of ordinary 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, and the like). Such impermissible substitution patterns are readily recognized by those of ordinary skill in the art. Any functional group disclosed herein and / or defined above may be substituted or unsubstituted, unless otherwise indicated herein. Unless otherwise explained, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. The singular terms "a," "an," and "the" include plural referred to unless the context clearly dictates otherwise. 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 weights or molecular mass values ​​given for nucleic acid or polypeptide are approximate and are provided for illustration purposes.Methods and materials similar or equivalent to those described herein can be used in carrying out or testing this disclosure, and suitable methods and materials are described herein.In case of discrepancies, the present specification shall prevail, including explanations of terms.In addition, materials, methods and examples are only illustrative and are not intended to be limiting. Description of the embodiment

[0125] Provided herein are compositions of particles comprising amphiphiles and drug molecules that are useful for treating or preventing diseases, such as cancer, autoimmune diseases, allergies, and / or infectious diseases. Particles comprising certain compositions of amphiphiles and peptide antigen conjugates are particularly useful for treating or preventing diseases, such as cancer, autoimmune diseases, allergies, and / or infectious diseases, as vaccines.

[0126] The present disclosure relates to a vaccine comprising an amphiphile having the formula S-[B]-[U]-H and at least one peptide antigen conjugate having a formula selected from [S]-[E1]-A-[E2]-[U]-H and H-[U]-[E1]-A-[E2]-[S], During the ceremony, S, independently for each occurrence, is a solubilizing block; B is a spacer, H, independently for each occurrence, is a hydrophobic block; one or more drug molecules (D) are optionally attached to each H, either directly or via a suitable linker X1; A is, independently for each occurrence, a peptide antigen; E1, independently for each occurrence, is an N-terminal extension, E2 is, independently for each occurrence, a C-terminal extension, U is, independently for each occurrence, a linker; [ ] 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; The amphiphile and / or at least one peptide antigen conjugate comprises a dendron amplifier.

[0127] In some embodiments of the vaccine, the amphiphile comprises a dendron amplifier, hi other embodiments, at least one peptide antigen conjugate comprises a dendron amplifier.

[0128] In some embodiments of the vaccine, the amphiphile S comprises a dendron amplifier, hi other embodiments, the amphiphile S has a dendritic structure.

[0129] In some vaccine embodiments, 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 dendron amplifiers, e.g., four to eight SGs are connected to S.

[0130] In some embodiments of the vaccine, the 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, phosphoserine and any derivative thereof, an agonist of CD22a, sialyl Lewis x, and combinations thereof.

[0131] In some vaccine embodiments, the dendron amplifier comprises 1 to 10 generations of repeating monomer units with 2 to 6 branches per generation. In other embodiments, the dendron amplifier comprises 2 to 3 generations of repeating monomer units with 2 to 3 branches per generation. In some vaccine embodiments, 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 for each occurrence selected from (CH2) y3 -FG2, (OCH2CH2) y3 -FG2, and CH2(OCH2CH2) y3-FG2), where y2 and y3, independently for each occurrence, are each an integer from 1 to 6 repeat units, FG1 is a first functional group, and FG2 is a second functional group. In some embodiments, FG1 is -NH2, and FG2, independently for each occurrence, is -CO2- or -CO2H. In some embodiments, FG1, independently for each occurrence, is -CO2- or -CO2H, and FG2 is -NH2.

[0132] In some embodiments of the vaccine, SG is linked to S via a suitable linker X5. In some embodiments of the vaccine, the suitable linker X5 linking SG to S is selected from lower alkyl and PEG groups. In some embodiments of the vaccine, two or more SGs are connected to the remainder of S by a dendron amplifier by a suitable linker X5 that links the two or more SGs to the terminal functional group (FGt) of the dendron amplifier by an amide bond. In some embodiments of the vaccine for inducing tolerance, the linker X5 that joins SG to the dendron amplifier is -NH-R 19 , -NH-C(O)-R 19 , -C(O)-NH-R 19 -or-C(O)-R 19 R in these formulas is selected from 19 is -(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, such as 1, 2, 3, 4, 5, or 6.

[0133] In some embodiments of the vaccine, the dendron amplifier comprises a polyethylene oxide (PEG) group.

[0134] 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.

[0135] 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.

[0136] 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.

[0137] In some embodiments of the vaccine, each H independently represents the formula: -(M) m -(N) N -(O) o -(P) p -R 3 and a poly(amino acid) having the formula: M, N, O, and P are each independently present or absent, provided that at least one of M or N is present; m, n, o, and p each independently represent an integer from 1 to 100, and the sum of m, n, o, and p is less than or equal to 100; R 3 is a hydrogen, NH, NH-CH, NH-(CH) connected either directly or by a suitable linker X y5 CH3, OH or a drug molecule (D); y5 is an integer selected from 1 to 6.

[0138] In some embodiments of the vaccine, P is absent. In other embodiments, N, O and P are each absent.

[0139] In some embodiments of the vaccine, P is [ka] where each R 5 are independently groups containing 1 to 2 charged functional groups.

[0140] 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.

[0141] 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 vaccine embodiments, 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 vaccine embodiments, X1 is present and comprises an enzyme-degradable peptide and / or a self-immolative linker.

[0142] In some embodiments, X is present and is —(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)-R7 , -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-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-CHR8 -C(O)) j -NH-C6H4-CH2-OC(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-OC(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 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 is selected from carboxylic acid, activated carboxylic acid (e.g., carbonylthiazolidine-2-thione ("TT"), NHS, or nitrophenol ester), carboxylic acid anhydride, amine and protected amine (e.g., tert-butyloxycarbonyl-protected amine), OSi(CH3), alkene, azide, alkyne, strained alkyne, halogen (e.g., fluoride, chloride), olefin and endocyclic olefin (e.g., allyl), CN, OH, and epoxy, hydrazine (including hydrazide), carbohydrazide, aldehyde, ketone, carbamate, and activated carbamate. where D is any suitable functional group for attachment to a functional group ("FG4") present on a drug molecule, including but not limited to, any suitable leaving group that may be selected from any suitable leaving group (e.g., NHS, TT, nitrophenol, etc.); w is a group resulting from either the reaction of FG4 with FG3 or the displacement of LG on FG4, and is typically selected from NH-, C(O)-, NH-C(O)-, C(O)-NH-, OC(O)-NH-, C(O)-NH-N=C(CH3)-, NH-N=C(CH3)-, or -C(CH3)=N-NH-C(O)-, and w is always linked to D either directly (i.e., w-D) or indirectly via X3 (i.e., w-X3-D).

[0143] In some embodiments of the vaccine, M is [ka] where each R 4 are independently hydrophobic groups.

[0144] In some embodiments of the vaccine, R 4 teeth, [ka] wherein: a is aryl or heteroaryl; X2 is present or absent and, if present, is a suitable linker; y8 is selected from an integer between 0 and 6; Z 1 , Z 2 , and Z 3 are each independently selected from H, F, hydroxy, amino, alkyl, and fluoroalkyl.

[0145] 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.

[0146] 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 wherein y9 in these formulas is typically an integer selected from 1 to 6. In other embodiments, X2 is present and is selected from a lower alkyl and a PEG group.

[0147] In some embodiments of the vaccine, each R 4 is, independently, [ka] [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, independently, [ka] [ka] wherein each y is independently selected from an integer from 0 to 6. In other embodiments, each R 4 is, independently, [ka] In other embodiments, each R 4 is, independently, [ka] [ka] In a preferred embodiment, each R 4 is, independently, [ka] is selected from.

[0148] In some embodiments of the vaccine, at least one D is [ka] wherein: 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.

[0149] 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.

[0150] In some embodiments of the vaccine, X 3 is alkyl.

[0151] In some embodiments of the vaccine, m, n, o, and p each independently represent an integer from 1 to 30, and the sum of m, n, o, and p is less than or equal to 30.

[0152] In some embodiments of the vaccine, m, n, o, and p each independently represent an integer from 1 to 10, and the sum of m, n, o, and p is less than or equal to 10.

[0153] 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.

[0154] 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.

[0155] 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.

[0156] 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.

[0157] In some vaccine embodiments, the vaccine comprises a molar ratio of peptide antigen conjugate to amphiphile of about 4:1 to about 1:20.

[0158] 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, a tolerance-inducing transplant rejection vaccine, a cardiovascular vaccine, or a neurodegenerative disease vaccine.

[0159] In some vaccine embodiments, the peptide antigen (A) comprises a sequence in which one or more cysteine ​​residues are replaced with alpha amino-butyric acid and / or one or more methionine residues are replaced with norleucine. In some vaccine embodiments, 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 or neurodegenerative diseases.

[0160] In some embodiments of the vaccine, A is selected from the group consisting of RGYLTKILHVFHGLLPGFLVKMSGDLLE, PGFLVKMSGDLLE, PGFLVKnSGDLLE (where n = norleucine); PGFLVKMSSDLLG, PGFLVKnSSDLLG (where n = norleucine); SIPWNLERITPPR; SIPWNLERITPPR; SIPWNLE; SIPWNLEKVTPPR; SIPWNLDRVTPPR; NVPEEDGTRFHRQASKC; NVPEEDGTRFHRQASK; PEEDGTR; NVPEEDG; NVPEEDATRFHRQGSK; LFAPGEDIIGASSDCSTCFVSQSGTSQAAA; CSTCFVSQSGTSQAAA; STCFVSQSGTSQAAA, STBFV SQSGTSQAAA;STBFVSQ;MFTIKLLLFIVPLVISSRIDQDNSSFDSLSPEPKSRFAMLDDVKILANGLLQLGHGLKDFVHKTKGQIND;EPKSRFAMLDDVKILANGLLQLGHGLKDFVHKTKGQIND;EPKSRFAMLDDVKILANGLLQLGHGLKDFVHKTKGQIND;EPKSRFAMLDDVKI;MLDDVKILANGLLQ;LANGLLQLGHGLKD;LGHGLKDFVHKTKG;LKDFVHKTKGQIND;RFAMLDDVKILANGLLQLGH;GLLQLGHGLKDFVHKTKGQI; and IFQKLNIFDQSFYDLSLQTSEIKEEEKELRRTTYKLQVKNEEVKNMSLELNSKLESLLEEKILLQQKVK.

[0161] In some embodiments of the vaccine, A is directly covalently linked to E1, which is covalently linked directly or indirectly via U to H.

[0162] In some embodiments of the vaccine, A is covalently linked directly to E2, which is covalently linked directly or indirectly via U to H.

[0163] In some embodiments of the vaccine, E1 and E2 each comprise a PEG group of 4 to 36 monomer units, for example, the PEG group comprises 4 to 24 monomer units.

[0164] In some embodiments of the vaccine, each of E1 and E2 comprises a peptide.

[0165] In some embodiments of the vaccine, the peptide comprises between 4 and 24 amino acids.

[0166] In some embodiments of the vaccine, the peptide comprises an amino acid selected from glycine, serine, threonine, alanine, and proline, e.g., the peptide is (Gly-Ser) 2~12 , (Gly-Gly-Gly-Gly-Ser) 1~4 , and (Ala-Pro) 2~12 is selected from.

[0167] In some embodiments of the vaccine, the peptide comprises between 7 and 28 amino acids.

[0168] In some embodiments of the vaccine, the peptide has the formula (AA H -AA P -AA P -AA H -AA P -AA P -AA P ) 1~4 wherein AA H is a hydrophobic amino acid suitable for a coil domain selected from isoleucine, leucine, valine and norleucine, and AAP is a hydrophilic amino acid suitable for the coil domain selected from alanine, serine, lysine, aspartic acid and glutamic acid, for example, the peptide is (Ile-Ala-Ala-Ile-Glu-Ser-Lys) 1~4 ,(Ile-Ala-Ala-Ile-Lys-Ser-Lys) 1~4 , and (Ile-Ala-Ala-Ile-Glu-Ser-Glu) 1~4 is selected from.

[0169] In some embodiments of the vaccine, at least one peptide-antigen conjugate comprises A selected from an autoantigen, an alloantigen, and an allergen.

[0170] 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, and N-acetylgalactosamine, and an agonist of CD22a.

[0171] In some embodiments of the vaccine, the vaccine comprises an inhibitor of mTOR, RORγt, 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.

[0172] In some embodiments of the vaccine, the vaccine comprises an inhibitor of mTOR, RORγt, CDK8 / 19 and HDAC, AHR, RAR and A 2a and a second drug molecule (D2) independently selected from an agonist of an NLR, a CLR, a TLR, and an immunostimulant selected from an agonist of STING, with the proviso that D and D2 bind to different receptors.

[0173] 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 inhibitor of mTOR, and D2 is selected from an agonist of NLR, CLR, TLR, and STING.

[0174] 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 an RNA and an imidazoquinoline agonist of TLR-7, TLR-8, and TLR-7 / 8.

[0175] In some embodiments of the vaccine, the vaccine comprises an inhibitor of mTOR, RORγt, CDK8 / 19 and HDAC, AHR, RAR and A 2a and a third drug molecule (D3) independently selected from an agonist of an NLR, a CLR, a TLR, and an immunostimulant selected from an agonist of STING, with the proviso that D, D2, and D3 bind to different receptors.

[0176] 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.

[0177] In some embodiments of the vaccine, the molar ratio of total peptide antigen conjugate to at least one D is from about 20:1 to 1:2, or from about 10:1 to about 1:1, or from about 4:1 to about 2:1.

[0178] In some embodiments of the vaccine, at least one peptide antigen conjugate comprises A selected from a tumor antigen.

[0179] 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.

[0180] In some vaccine embodiments, each H of the amphiphile and / or peptide antigen conjugate independently comprises a poly(amino acid) comprising a monomer of a hydrophobic amino acid (M) selected from tryptophan, 1-methyltryptophan, and / or para-aminophenylalanine. In other embodiments, 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 a D selected from an agonist of TLR-7 / 8. In some vaccine embodiments, 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 vaccine embodiments, 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 from about 20:1 to 1:2, or from about 10:1 to about 1:1, or from about 4:1 to about 2:1.

[0181] In some embodiments of the vaccine, A is a glycopeptide. In other embodiments, A is selected from HGVT*S*APDT*RPAPGS*T*APPA, DT*RPAPGS*T*APPAHGVT*S*AP, GS*T*APPAHGVT*S*APDT*RPAPGS*T*APPA, GVT*S*APDT*RPAP, APDT*RPAPGS*T*A, GS*T*APPAHGVT*S*AP, VT*S*AP, DT*RPAP, and GS*T*AP, where * 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, sTF.

[0182] 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.

[0183] 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).

[0184] In some embodiments of the vaccine, B comprises 4-36 monomer units and the amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0185] 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).

[0186] 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.

[0187] In some vaccine embodiments, B comprises 4 to 36 PEG monomer units and the amphiphile H comprises a polymer of para-amino-phenylalanine.

[0188] 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.

[0189] 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.

[0190] In some vaccine embodiments, B comprises 4-36 PEG monomer units and the amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and a reactive amino acid (N) comprising an imidazoquinoline.

[0191] 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) comprising an imidazoquinoline.

[0192] 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) with reactive amino acids (N) including tryptophan and imidazoquinoline.

[0193] In some vaccine embodiments, B comprises 4-36 PEG monomer units and the amphiphile H comprises a poly(amino acid) with reactive amino acids (N) including tryptophan and imidazoquinoline.

[0194] 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) with reactive amino acids (N) including tryptophan and imidazoquinoline.

[0195] 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 SG comprises mannose.

[0196] 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.

[0197] In some vaccine embodiments, B comprises 4-36 PEG monomer units, the amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M), and SG comprises mannose.

[0198] 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.

[0199] 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.

[0200] In some embodiments of the vaccine, B comprises 4-36 PEG monomer units, the amphiphile H comprises a polymer of para-amino-phenylalanine, and SG comprises mannose.

[0201] 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.

[0202] 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 SG comprises mannose.

[0203] 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) comprising an imidazoquinoline, and SG comprises mannose.

[0204] In some embodiments of the vaccine, amphiphile S comprises a second or third generation dendrimer, B comprises 4-36 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.

[0205] In some embodiments of the vaccine, the amphiphile S comprises a second or third generation dendrimer, the amphiphile H comprises a poly(amino acid) with reactive amino acids (N) including tryptophan and imidazoquinoline, and the amphiphile SG comprises mannose.

[0206] In some embodiments of the vaccine, B comprises 4-36 PEG monomer units, the amphiphile H comprises a poly(amino acid) with reactive amino acids (N) including tryptophan and imidazoquinoline, and SG comprises mannose.

[0207] 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) with tryptophan and a reactive amino acid (N) comprising an imidazoquinoline, and SG comprises mannose.

[0208] In some vaccine embodiments, the amphiphile S comprises a second or third generation dendrimer, B comprises 4-36 PEG monomer units, and the peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0209] 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).

[0210] In some vaccine embodiments, B comprises 4-36 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).

[0211] In some embodiments of the vaccine, the amphiphile S comprises a second or third generation dendrimer, B comprises 4-36 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).

[0212] 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).

[0213] In some vaccine embodiments, 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).

[0214] In some vaccine embodiments, the amphiphile S comprises a second or third generation dendrimer, B comprises 4-36 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).

[0215] 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).

[0216] In some vaccine embodiments, 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) comprising an imidazoquinoline, and the peptide-antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0217] In some vaccine embodiments, the amphiphile S comprises a second or third generation dendrimer, 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) comprising an imidazoquinoline, and the peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0218] In some vaccine embodiments, the amphiphile S comprises a second or third generation dendrimer, the amphiphile H comprises a poly(amino acid) with 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).

[0219] In some vaccine embodiments, B comprises 4-36 PEG monomer units, the amphiphile H comprises a poly(amino acid) with reactive amino acids (N) including tryptophan and imidazoquinoline, and the peptide antigen conjugate H comprises a poly(amino acid) with a hydrophobic amino acid (M).

[0220] In some vaccine embodiments, the amphiphile S comprises a second or third generation dendrimer, B comprises 4-36 PEG monomer units, the amphiphile H comprises a poly(amino acid) with 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).

[0221] In some vaccine embodiments, the amphiphile S comprises a second or third generation dendrimer, B comprises 4-36 PEG monomer units, SG comprises mannose, and H of the peptide antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0222] 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).

[0223] 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), SG comprises mannose, and the peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0224] In some embodiments of the vaccine, amphiphile S comprises a second or third generation dendrimer, B comprises 4-36 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).

[0225] 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).

[0226] In some vaccine embodiments, B comprises 4-36 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).

[0227] In some embodiments of the vaccine, amphiphile S comprises a second or third generation dendrimer, B comprises 4-36 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).

[0228] 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, the SG comprises mannose, and the peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0229] In some vaccine embodiments, 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) comprising an imidazoquinoline, SG comprises mannose, and the peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0230] In some embodiments of the vaccine, amphiphile S comprises a second or third generation dendrimer, B comprises 4-36 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 mannose, and peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0231] In some embodiments of the vaccine, the amphiphile S comprises a second or third generation dendrimer, the amphiphile H comprises a poly(amino acid) with reactive amino acids (N) comprising tryptophan and imidazoquinoline, the SG comprises mannose, and the peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0232] In some vaccine embodiments, B comprises 4-36 PEG monomer units, the amphiphile H comprises a poly(amino acid) with 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).

[0233] In some vaccine embodiments, the amphiphile S comprises a second or third generation dendrimer, B comprises 4-36 PEG monomer units, the amphiphile H comprises a poly(amino acid) with 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).

[0234] 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).

[0235] 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).

[0236] In some embodiments of the vaccine, B comprises 4-36 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).

[0237] In some embodiments of the vaccine, the amphiphile S comprises a second or third generation dendrimer, B comprises 4-36 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).

[0238] 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).

[0239] In some embodiments of the vaccine, B comprises 4-36 PEG monomer units, H of the amphiphile comprises a polymer of para-amino-phenylalanine, the amphiphile comprises amino-hexanoic acid, and H of the peptide antigen conjugate 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, 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).

[0241] 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, the amphiphile comprises an amino-hexanoic acid, and the peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0242] In some embodiments of the vaccine, B comprises 4-36 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).

[0243] In some embodiments of the vaccine, the amphiphile S comprises a second or third generation dendrimer, 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) 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).

[0244] In some embodiments of the vaccine, the amphiphile S comprises a second or third generation dendrimer, the amphiphile H comprises a poly(amino acid) with 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).

[0245] In some vaccine embodiments, B comprises 4-36 PEG monomer units, the amphiphile H comprises a poly(amino acid) with 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) with a hydrophobic amino acid (M).

[0246] In some vaccine embodiments, the amphiphile S comprises a second or third generation dendrimer, B comprises 4-36 PEG monomer units, the amphiphile H comprises a poly(amino acid) with 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).

[0247] In some vaccine embodiments, the amphiphile S comprises a second or third generation dendrimer, B comprises 4-36 PEG monomer units, the dendrimer monomers comprise hydroxy acids and amino alcohols, and the peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0248] 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).

[0249] In some vaccine embodiments, 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).

[0250] In some vaccine embodiments, the amphiphile S comprises a second or third generation dendrimer, 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).

[0251] 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).

[0252] In some vaccine embodiments, 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).

[0253] In some vaccine embodiments, the amphiphile S comprises a second or third generation dendrimer, 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).

[0254] 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, 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).

[0255] In some vaccine embodiments, 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).

[0256] In some vaccine embodiments, the amphiphile S comprises a second or third generation dendrimer, 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).

[0257] In some vaccine embodiments, the amphiphile S comprises a second or third generation dendrimer, the amphiphile H comprises a poly(amino acid) with 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) with a hydrophobic amino acid (M).

[0258] In some vaccine embodiments, B comprises 4-36 PEG monomer units, the amphiphile H comprises a poly(amino acid) with 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) with a hydrophobic amino acid (M).

[0259] In some vaccine embodiments, the amphiphile S comprises a second or third generation dendrimer, B comprises 4-36 PEG monomer units, the amphiphile H comprises a poly(amino acid) with reactive amino acids (N) including tryptophan and imidazoquinoline, the dendrimer monomers comprise hydroxy acids and amino alcohols, and the peptide antigen conjugate H comprises a poly(amino acid) with a hydrophobic amino acid (M).

[0260] The present disclosure also relates to a vaccine for inducing tolerance comprising an amphiphile having the formula S-[B]-[U]-H and at least one peptide antigen conjugate having a formula selected from [S]-[E1]-A-[E2]-[U]-H and H-[U]-[E1]-A-[E2]-[S], During the ceremony, S, independently for each occurrence, is a solubilizing block; B is a spacer, H, independently for each occurrence, is a hydrophobic block; one or more drug molecules (D) are optionally attached to each H, either directly or via a suitable linker X1; A is, independently for each occurrence, a peptide antigen; E1, independently for each occurrence, is an N-terminal extension, E2 is, independently for each occurrence, a C-terminal extension, U is, independently for each occurrence, a linker; [ ] 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; The amphiphile and / or the at least one peptide antigen conjugate comprises a dendron amplifier, and the at least one peptide antigen is selected from an autoantigen, an alloantigen, and an allergen.

[0261] 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.

[0262] In some embodiments of the vaccine for inducing tolerance, the amphiphile comprises a dendron amplifier, hi other embodiments, at least one peptide-antigen conjugate comprises a dendron amplifier.

[0263] In some embodiments of the vaccine for inducing tolerance, the amphiphile S comprises a dendron amplifier, hi other embodiments, the amphiphile S has a dendritic structure.

[0264] 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 dendron amplifiers, e.g., four to eight SGs are connected to the S.

[0265] In some embodiments of the vaccine for inducing tolerance, the 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, phosphoserine and any derivative thereof, an agonist of CD22a, sialyl Lewis x, and combinations thereof.

[0266] 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.

[0267] 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.

[0268] In some embodiments of a 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 for each occurrence, (CH2) y3 -FG2, (OCH2CH2) y3 -FG2, and CH2(OCH2CH2) y3 -FG2), where y2 and y3, independently for each occurrence, are integers from 1 to 6 repeat units, FG1 is a first functional group, and FG2 is a second functional group. In some embodiments, FG1 is -NH2, and FG2, independently for each occurrence, is -CO2- or -CO2H. In some embodiments, FG1 is -CO2- or -CO2H, and FG2, independently for each occurrence, is -NH2.

[0269] 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 a dendron amplifier by a suitable linker X5 that links the two or more SGs to the terminal functional group (FGt) of the dendron amplifier by an amide bond. In some embodiments of the vaccine for inducing tolerance, the linker X5 that joins SG to the dendron amplifier is -NH-R 19 , -NH-C(O)-R 19 , -C(O)-NH-R 19 -or-C(O)-R 19 R in these formulas is selected from 19 is -(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, such as 1, 2, 3, 4, 5, or 6.

[0270] In some embodiments of the vaccine for inducing tolerance, the dendron amplifier comprises a polyethylene oxide (PEG) group.

[0271] 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.

[0272] In some embodiments of the vaccine for inducing tolerance, H of the peptide-antigen conjugate comprises a higher alkane, an aromatic group, a fatty acid, a sterol, a polyunsaturated hydrocarbon, and / or a polymer.

[0273] 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.

[0274] In some embodiments of the vaccine for inducing tolerance, each H independently represents the formula: -(M) m -(N) N -(O) o -(P) p -R 3 and a poly(amino acid) having the formula: M, N, O, and P are each independently present or absent, provided that at least one of M or N is present; m, n, o, and p each independently represent an integer from 1 to 100, and the sum of m, n, o, and p is less than or equal to 100; R 3 is a hydrogen, NH, NH-CH, NH-(CH) connected either directly or by a suitable linker X y5 CH3, OH or a drug molecule (D); y5 is an integer selected from 1 to 6.

[0275] In some embodiments of the vaccine for inducing tolerance, P is absent. In other embodiments, N, O and P are each absent.

[0276] In some embodiments of the vaccine for inducing tolerance, P is [ka] where each R 5 are independently groups containing 1 to 2 charged functional groups.

[0277] 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.

[0278] In some embodiments of the vaccine for inducing tolerance, N is [ka] wherein each X is independently a suitable linker and each D is independently a drug molecule.

[0279] In some embodiments of the vaccine for inducing tolerance, M is [ka] where each R 4 are independently hydrophobic groups.

[0280] In some embodiments of the vaccine for inducing tolerance, R 4 teeth, [ka] wherein: a is aryl or heteroaryl; X2 is present or absent and, if present, is a suitable linker; Y8 is selected from an integer from 0 to 6; Z 1 , Z 2 , and Z3 are each independently selected from H, F, hydroxy, amino, alkyl, and fluoroalkyl.

[0281] 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.

[0282] 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 wherein y9 in these formulas is typically an integer selected from 1 to 6. In other embodiments, X2 is present and is selected from a lower alkyl and a PEG group.

[0283] In some embodiments of a vaccine for inducing tolerance, each R 4 is, independently, [ka] [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, independently, [ka] [ka] wherein each y is independently selected from an integer from 0 to 6. In other embodiments, each R 4 is, independently, [ka] In other embodiments, each R 4 is, independently, [ka] [ka] [ka] In a preferred embodiment, each R 4 is, independently, [ka] wherein y is selected from integers of 1 to 6.

[0284] In some embodiments of a vaccine for inducing tolerance, at least one D is [ka] wherein: R 20 is selected from H, alkyl, alkoxyalkyl, aryl, heteroaryl, aminoalkyl, amide, and ester; X3 is selected from alkyl, alkoxyalkyl, aralkyl, heteroaralkyl, aryl, heteroaryl, and carboxy.

[0285] In some embodiments of the vaccine for inducing tolerance, R 20 is selected from H, alkyl and alkoxyalkyl, and X is selected from alkyl and aralkyl. 20 is butyl.

[0286] In some embodiments of the vaccine for inducing tolerance, X3 is alkyl.

[0287] In some embodiments of the vaccine for inducing tolerance, m, n, o, and p each independently represent an integer from 1 to 30, and the sum of m, n, o, and p is less than or equal to 30.

[0288] In some embodiments of the vaccine for inducing tolerance, m, n, o, and p each independently represent an integer from 1 to 10, and the sum of m, n, o, and p is less than or equal to 10.

[0289] 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.

[0290] 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.

[0291] 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.

[0292] 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.

[0293] 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.

[0294] 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 amino-butyric acid and / or one or more methionine residues are replaced with norleucine.

[0295] In some embodiments of a vaccine for inducing tolerance, the vaccine comprises an inhibitor of mTOR, RORγt, 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.

[0296] In some embodiments of a vaccine for inducing tolerance, the vaccine comprises an inhibitor of mTOR, RORγt, CDK8 / 19 and HDAC, AHR, RAR and A 2a and a second drug molecule (D2) independently selected from an agonist of an NLR, a CLR, a TLR, and an immunostimulant selected from an agonist of STING, with the proviso that D and D2 bind to different receptors.

[0297] 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.

[0298] In some embodiments of the vaccine for inducing tolerance, at least one D is selected from an ATP-competitive inhibitor of mTOR and D2 is selected from an agonist of an NLR, a CLR, a TLR, and a STING.

[0299] In some 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 some embodiments of the vaccine for inducing tolerance, D2 is selected from RNA and imidazoquinoline agonists of TLR-7, TLR-8, and TLR-7 / 8.

[0300] In some embodiments of a vaccine for inducing tolerance, the vaccine comprises an inhibitor of mTOR, RORγt, CDK8 / 19 and HDAC, AHR, RAR and A 2aand a third drug molecule (D3) independently selected from an agonist of an NLR, a CLR, a TLR, and an immunostimulant selected from an agonist of STING, with the proviso that D, D2, and D3 bind to different receptors.

[0301] In some embodiments of the 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.

[0302] 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.

[0303] 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.

[0304] 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).

[0305] In some embodiments of the vaccine for inducing tolerance, B comprises 4-36 PEG monomer units and the amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0306] 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, and amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0307] 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.

[0308] 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.

[0309] 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, and amphiphile H comprises a polymer of para-amino-phenylalanine.

[0310] 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) and a reactive amino acid (N) comprising an imidazoquinoline.

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

[0312] 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) comprising a hydrophobic amino acid (M) and a reactive amino acid (N) comprising an imidazoquinoline.

[0313] 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) with reactive amino acids (N) including tryptophan and imidazoquinoline.

[0314] In some embodiments of the vaccine for inducing tolerance, B comprises 4-36 PEG monomer units and the amphiphile H comprises a poly(amino acid) with reactive amino acids (N) including tryptophan and imidazoquinoline.

[0315] 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) with reactive amino acids (N) including tryptophan and imidazoquinoline.

[0316] 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.

[0317] 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 SG comprises N-acetylgalactosamine.

[0318] 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 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 B comprises 4 to 36 PEG monomer units, the amphiphile H comprises a poly(amino acid) comprising a hydrophobic amino acid (M), and the amphiphile SG comprises N-acetylgalactosamine.

[0320] 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.

[0321] 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, and SG comprises N-acetylgalactosamine.

[0322] 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.

[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 a reactive amino acid (N) comprising an imidazoquinoline, and the amphiphile SG comprises N-acetylgalactosamine.

[0324] 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) comprising an imidazoquinoline, and SG comprises N-acetylgalactosamine.

[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 a reactive amino acid (N) comprising an imidazoquinoline, and the amphiphile SG comprises N-acetylgalactosamine.

[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 poly(amino acid) with reactive amino acids (N) including tryptophan and imidazoquinoline, and the amphiphile SG comprises N-acetylgalactosamine.

[0327] In some embodiments of the vaccine for inducing tolerance, B comprises 4-36 PEG monomer units, the amphiphile H comprises a poly(amino acid) with reactive amino acids (N) including tryptophan and imidazoquinoline, and SG comprises N-acetylgalactosamine.

[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 poly(amino acid) with tryptophan and a reactive amino acid (N) comprising an imidazoquinoline, and the amphiphile SG comprises N-acetylgalactosamine.

[0329] 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.

[0330] 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.

[0331] 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.

[0332] 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 comprises an enzymatically degradable linker.

[0333] 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.

[0334] 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.

[0335] 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.

[0336] 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.

[0337] 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) comprising an imidazoquinoline, and the peptide-antigen conjugate comprises an enzyme-degradable linker.

[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 a reactive amino acid (N) comprising an imidazoquinoline, and the peptide-antigen conjugate comprises an enzyme-degradable linker.

[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 poly(amino acid) with reactive amino acids (N) including tryptophan and imidazoquinoline, and the peptide antigen conjugate comprises an enzyme-degradable linker.

[0340] 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, and the peptide-antigen conjugate comprises an enzyme-degradable linker.

[0341] 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) with reactive amino acids (N) including tryptophan and imidazoquinoline, and the peptide antigen conjugate comprises an enzyme-degradable linker.

[0342] 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 H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0343] 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 H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0344] 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).

[0345] 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).

[0346] 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).

[0347] 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, and the peptide antigen conjugate H comprises a poly(amino acid) comprising 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, 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).

[0349] 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 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, 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).

[0351] 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).

[0352] 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) with 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).

[0353] In some embodiments of the vaccine for inducing tolerance, B comprises 4-36 PEG monomer units, the amphiphile H comprises a poly(amino acid) with 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).

[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 poly(amino acid) with 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).

[0355] 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).

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

[0357] 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), SG comprises N-acetylgalactosamine, and the peptide-antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0358] 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), SG comprises N-acetylgalactosamine, and the peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0359] 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, the SG comprises N-acetylgalactosamine, and the peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0360] 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).

[0361] 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).

[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 a reactive amino acid (N) comprising an imidazoquinoline, the SG comprises N-acetylgalactosamine, 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 4-36 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 SG comprises N-acetylgalactosamine, and the peptide-antigen conjugate H 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 H comprises a poly(amino acid) comprising a hydrophobic amino acid (M) and a reactive amino acid (N) comprising an imidazoquinoline, the SG comprises N-acetylgalactosamine, and the peptide-antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0365] 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) with reactive amino acids (N) comprising tryptophan and imidazoquinoline, the SG comprises N-acetylgalactosamine, 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 4-36 PEG monomer units, the amphiphile H comprises a poly(amino acid) with 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).

[0367] 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) with 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).

[0368] 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).

[0369] 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).

[0370] 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).

[0371] 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).

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

[0373] 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), the amphiphile comprises amino-hexanoic acid, and H of the peptide-antigen conjugate comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0374] 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), the amphiphile comprises amino-hexanoic acid, and the peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0375] 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, the amphiphile comprises amino-hexanoic acid, and the peptide antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0376] 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 amphiphile comprises amino-hexanoic acid, and the peptide antigen conjugate H comprises a poly(amino acid) comprising 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 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).

[0378] 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, the amphiphile comprises an amino-hexanoic acid, 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) 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).

[0380] 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, the amphiphile comprises amino-hexanoic acid, and the peptide-antigen conjugate H comprises a poly(amino acid) comprising a hydrophobic amino acid (M).

[0381] 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) with reactive amino acids (N) comprising tryptophan and imidazoquinoline, the amphiphile comprises amino-hexanoic acid, 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 poly(amino acid) with 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) with a hydrophobic amino acid (M).

[0383] In some embodiments of the vaccine for inducing tolerance, the amphiphile S comprises a second or third generation dendrimer, B comprises 4-36 PEG monomer units, the amphiphile H comprises a poly(amino acid) with 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).

[0384] 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).

[0385] 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), 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 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).

[0387] 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), 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).

[0388] 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, 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).

[0389] 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).

[0390] 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, 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).

[0391] 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, 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).

[0392] 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).

[0393] 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) 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).

[0394] 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) with 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).

[0395] In some embodiments of the vaccine for inducing tolerance, B comprises 4-36 PEG monomer units, the amphiphile H comprises a poly(amino acid) with 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) with a hydrophobic amino acid (M).

[0396] 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) with reactive amino acids (N) including tryptophan and imidazoquinoline, the dendrimer monomers comprise hydroxy acids and amino alcohols, and the peptide antigen conjugate H comprises a poly(amino acid) including a hydrophobic amino acid (M).

[0397] The present disclosure also relates to a vaccine comprising an amphiphile having the formula S-[B]-[U]-H and at least one peptide antigen conjugate having a formula selected from [S]-[E1]-A-[E2]-[U]-H and H-[U]-[E1]-A-[E2]-[S], wherein S is, independently for each occurrence, a solubilizing block; B is a spacer, H, independently for each occurrence, is a hydrophobic block; one or more drug molecules (D) are optionally attached to each H, either directly or via a suitable linker X1; A is, independently for each occurrence, a peptide antigen; E1, independently for each occurrence, is an N-terminal extension, E2 is, independently for each occurrence, a C-terminal extension, U is, independently for each occurrence, a linker; [ ] indicates that the group is optional; - indicates that two adjacent groups are covalently linked to each other directly or indirectly via a suitable linker X; the amphiphile and / or at least one peptide-antigen conjugate comprises a dendron amplifier; At least one A comprises a sequence in which one or more cysteine ​​residues are replaced with alpha amino-butyric acid and / or one or more methionine residues are replaced with norleucine.

[0398] In some embodiments of the vaccine, S is present in at least one peptide-antigen conjugate and comprises an SG selected from an amine or a carboxylic acid. In some embodiments of the vaccine, S comprises one or more lysine or ornithine residues.

[0399] In some embodiments of the vaccine, the peptide-antigen conjugate has a net positive charge of about +1 to about +10 at physiological pH.

[0400] In some embodiments of the vaccine, the peptide-antigen conjugate has a net positive charge of about +2 to about +6 or about +3 to about +5 at physiological pH.

[0401] In some embodiments of the vaccine, S comprises one or more glutamic acid or aspartic acid residues.

[0402] In some embodiments of the vaccine, the peptide-antigen conjugate has a net negative charge of about -1 to about -10 at physiological pH. In some embodiments of the vaccine, the peptide-antigen conjugate has a net negative charge of about -2 to about -6 or about -3 to about -5 at physiological pH.

[0403] In some embodiments of the vaccine, the amphiphile S comprises a carboxylic acid. In some embodiments of the vaccine, the amphiphile S comprises succinic acid or beta-alanine.

[0404] In some embodiments of the vaccine, the molar ratio of peptide antigen conjugate to amphiphile is about 4:1 to 1:20.

[0405] 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 from about 4:1 to about 2:1, or from about 1:2 to about 1:16, or from about 1:2 to about 1:4.

[0406] The present disclosure also relates to a vaccine 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 amino-butyric acid and / or one or more methionine residues are replaced with norleucine.

[0407] 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.

[0408] The present disclosure also relates to a vaccine comprising at least one peptide-antigen conjugate having a formula selected from [S]-[E1]-A-[E2]-[U]-H and H-[U]-[E1]-A-[E2]-[S], During the ceremony, S, independently for each occurrence, is a solubilizing block; H, independently for each occurrence, is a hydrophobic block; one or more drug molecules (D) are optionally attached to each H, either directly or via a suitable linker X1; A is, independently for each occurrence, a peptide antigen; E1, independently for each occurrence, is an N-terminal extension, E2 is, independently for each occurrence, a C-terminal extension, U is, independently for 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, and at least one D is present and selected from an agonist of TLR-7 / 8, and the vaccine further comprises a second drug molecule (D2) selected from an inhibitor of mTOR; (iii) at least one A is a glycopeptide; or (iv) at least one A is selected from an autoantigen, an allergen, and an alloantigen, and at least one D is present and selected from an ATP-competitive mTOR inhibitor; Either [ ] 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.

[0409] In some embodiments of the vaccine, at least one peptide antigen conjugate comprises at least one A selected from a tumor antigen.

[0410] 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.

[0411] In some embodiments of the vaccine, each H of the amphiphile and / or peptide antigen conjugate comprises a poly(amino acid) comprising monomers of reactive amino acids (N), wherein the monomers comprise D selected from an agonist of TLR-7 / 8.

[0412] In some embodiments of the vaccine, D2 is selected from rapamycin, tacrolimus, everolimus, RAD001, CCI-779, and AP23573.

[0413] In some embodiments of the vaccine, the molar ratio of peptide antigen conjugate to D2 is from about 20:1 to 1:2, or from about 10:1 to about 1:1, or from about 4:1 to about 2:1.

[0414] In some embodiments of the vaccine, at least one A is a glycopeptide, e.g., A is a glycopeptide selected from HGVT*S*APDT*RPAPGS*T*APPA, DT*RPAPGS*T*APPAHGVT*S*AP, GS*T*APPAHGVT*S*APDT*RPAPGS*T*APPA, GVT*S*APDT*RPAP, APDT*RPAPGS*T*A, GS*T*APPAHGVT*S*AP, VT*S*AP, DT*RPAP and GS*T*AP, where * 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, sTF.

[0415] In some embodiments, S is absent. In other embodiments, S is present.

[0416] In some embodiments of the vaccine, the vaccine further comprises an amphiphile having the formula S-[B]-[U]-H, During the ceremony, S is a solubilizing block, B is a spacer, H is a hydrophobic block, U is a linker, [ ] 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; The amphiphile S contains a dendron amplifier.

[0417] 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.

[0418] 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.

[0419] In some embodiments of the vaccine, the vaccine comprises an inhibitor of mTOR, RORγt, 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

[0420] In some embodiments of the vaccine, the vaccine comprises an inhibitor of mTOR, RORγt, CDK8 / 19 and HDAC, AHR, RAR and A 2a and a second drug molecule (D2) independently selected from an agonist of an NLR, a CLR, a TLR, and an immunostimulant selected from an agonist of STING, with the proviso that D and D2 bind to different receptors.

[0421] In some embodiments of the vaccine, D2 is selected from an agonist of NLR, CLR, TLR, and 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 an RNA and an imidazoquinoline agonist of TLR-7, TLR-8, and TLR-7 / 8.

[0422] In some embodiments of the vaccine, the vaccine comprises an inhibitor of mTOR, RORγt, CDK8 / 19 and HDAC, AHR, RAR and A 2aand a third drug molecule (D3) independently selected from an agonist of an NLR, a CLR, a TLR, and an immunostimulant selected from an agonist of STING, with the proviso that D, D2, and D3 bind to different receptors.

[0423] 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.

[0424] In some embodiments of the vaccine, the molar ratio of total peptide antigen conjugate to at least one D is from about 20:1 to 1:2, or from about 10:1 to about 1:1, or from about 4:1 to about 2:1.

[0425] In some embodiments of the vaccine, the vaccine further comprises an amphiphile having the formula S-[B]-[U]-H, During the ceremony, S is a solubilizing block, B is a spacer, H is a hydrophobic block, U is a linker, [ ] 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; The amphiphile S contains a dendron amplifier.

[0426] 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.

[0427] In some embodiments of the vaccine, S of at least one peptide antigen conjugate comprises SG selected from amines, hi other embodiments, S of the peptide antigen conjugate comprises one or more lysine or ornithine residues.

[0428] In some vaccine embodiments, 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.

[0429] 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.

[0430] 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.

[0431] In some vaccine embodiments, 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.

[0432] The present disclosure also relates to a vaccine comprising an expression system comprising DNA or RNA encoding at least one peptide antigen (A), further comprising at least one drug molecule (D) selected from a Treg-promoting immunomodulator.

[0433] In some embodiments of the vaccine comprising the expression system, at least one D is selected from an ATP-competitive mTOR inhibitor.

[0434] In some embodiments of the vaccine comprising the expression system, 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.

[0435] In some embodiments of the vaccine comprising the expression system, the peptide antigen (A) is selected from an autoantigen, an alloantigen, and an allergen.

[0436] In some embodiments of the vaccine comprising the expression system, the vaccine further comprises cationic liposome particles.

[0437] 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 [S]-[E1]-A-[E2]-[U]-H or H-[U]-[E1]-A-[E2]-[S], where 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, [ ] indicates that the group is optional, - indicates that two adjacent groups are covalently linked to each other directly or indirectly via a suitable linker X, and S, U and H of the amphiphile and the peptide antigen conjugate may be the same or different and may comprise one or more of the same functional groups or moieties.

[0438] In some vaccine embodiments, the amphiphile and / or peptide antigen conjugate further comprises one or more drug molecules (D), which may be linked either 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 [S]-[E1]-A-[E2]-[U]-HD). The drug molecule (D) may be mixed with the amphiphile and / or peptide antigen conjugate (e.g., D + S-[B]-[U]-H + [S]-[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), which is mixed with the amphiphile and / or peptide antigen conjugate (e.g., D-H + ​​S-[B]-[U]-H + [S]-[E1]-A-[E2]-[U]-H). Preferred compositions of vaccines further comprising a drug molecule (D) are described throughout this specification. D is attached directly or indirectly as a side chain of an adjacent group or as part of a side chain group.

[0439] In a preferred embodiment of the vaccine, the vaccine comprises particles comprising an amphiphile and one or more peptide antigen conjugates, which further comprise a drug molecule (D) selected from immunomodulators. A drug molecule (D) selected from an immunomodulatory agent may be linked to the hydrophobic block (H) of the amphiphile and / or peptide antigen conjugate either directly or indirectly via X1 (e.g., S-[B]-[U]-HD and / or [S]-[E1]-A-[E2]-[U]-HD); the drug molecule (D) may be mixed with the amphiphile and peptide antigen conjugate (e.g., D + S-[B]-[U]-H + [S]-[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), which is mixed with the amphiphile and peptide antigen conjugate (e.g., D-H + ​​S-[B]-[U]-H + [S]-[E1]-A-[E2]-[U]-H). Preferred compositions of vaccines further comprising a drug molecule (D) are described elsewhere herein.

[0440] 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 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 or preventing infectious diseases, the peptide antigen conjugate comprises an antigen (A) selected from a virus, bacterium, protozoan, or fungus. In still other embodiments, the vaccine comprises an antigen selected from an endogenously produced protein, and the vaccine is used to treat cardiovascular disease. In yet other embodiments of the vaccine, the antigen is selected from a small molecule hapten, and the vaccine is used to prevent toxicity upon exposure to chemical toxins, including nerve agents. Preferred antigens for treating different diseases, as well as preferred methods for selecting antigens for treating different diseases, are described throughout this specification.

[0441] Particles comprising certain compositions of amphiphiles have been found to be particularly useful for the delivery of small molecule drugs for a variety of applications, including the treatment of cancer, inflammation, autoimmune diseases, macular degeneration, and diseases of vital organs including the CNS, heart and liver, and metabolic diseases.

[0442] 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 drug and / or an immunomodulator. 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, and [ ] indicates an optional group, and the drug, D, is associated with the particle by covalent or non-covalent interactions.

[0443] The present disclosure also relates to a peptide antigen conjugate having a formula selected from S-[E1]-A-[E2]-[U]-H-[D] and [D]-H-[U]-[E1]-A-[E2]-S, or a peptide antigen fragment having a formula selected from S-[E1]-A-[E2]-[U1] and [U1]-[E1]-A-[E2]-S. In some embodiments, a drug molecule (D) is linked to the hydrophobic block (H) of the amphiphile, e.g., S-[B]-[U]-HD, where one or more Ds are attached to adjacent groups either directly or indirectly via X1, either at the terminus or as part of a side group. In other embodiments, a drug molecule is mixed with the amphiphile (e.g., D + S-[B]-[U]-H), or linked to the hydrophobic block (H) and mixed with the amphiphile (e.g., D-[B]-[U]-H + S-[B]-[U]-H, or H-D + S-[B]-[U]-H), and the drug is incorporated into particles formed by the amphiphile. Preferred compositions for cancer treatment comprising an amphiphile and at least one chemotherapeutic and / or immunostimulatory agent are described throughout this specification.

[0444] The present disclosure also relates to a peptide antigen conjugate having a formula selected from S-[E1]-A-[E2]-[U]-H-[D] and [D]-H-[U]-[E1]-A-[E2]-S, or a peptide antigen fragment having a formula selected from S-[E1]-A-[E2]-[U1] and [U1]-[E1]-A-[E2]-S, During the ceremony, S is a solubilizing block, H is a hydrophobic block, one or more drug molecules (D) are optionally attached to each H, either 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, [ ] indicates that the group is optional; - indicates that two adjacent groups are covalently linked to each other directly or indirectly via a suitable linker X; S comprises one or more amino acids.

[0445] In some embodiments, S comprises 2 to 12 amino acids. In some other embodiments, S comprises 2 to 8 amino acids. In some other embodiments, S comprises 4 to 6 amino acids. In some other embodiments, S comprises 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 amino acids.

[0446] In some embodiments, the S amino acids are selected from lysine, arginine, and ornithine.

[0447] In some embodiments, the peptide antigen fragment has the formula A-[E2]-S.

[0448] In some embodiments, E1 and / or E2 are present and are selected from cathepsin-cleavable tetrapeptides of the formula P4-P3-P2-P1.

[0449] In some embodiments, E1 and / or E2 are present and selected from Ser-Pro-Val-Arg, Ser-Pro-Val-Cit, and Ser-Pro-Val-aBut.

[0450] In some embodiments, the peptide antigen (A) comprises at least one amino acid selected from norleucine and alpha-aminobutyric acid.

[0451] In some embodiments, a vaccine comprising a peptide-antigen conjugate or peptide-antigen fragment disclosed herein.

[0452] In some embodiments, a method of activating, priming and / or expanding T cells, comprising adding to T cells in vitro or ex vivo an aqueous solution comprising a peptide-antigen conjugate or peptide-antigen fragment disclosed herein.

[0453] In some embodiments, at least one of E1 and E2 is present in a peptide antigen conjugate or peptide antigen fragment.

[0454] In some embodiments, each of E1 and / or E2 independently represents a group of formula (AA a -AA b -AA c -AA d -AA e -AA f -AA g ) e wherein: AA a and A.A. d is independently selected from leucine, isoleucine, norleucine, valine, norvaline, T-leucine, alloisoleucine, N-propylglycine, methionine, and O-methylserine; AA b , A.A. c and A.A. f is independently selected from alanine, cysteine, aspartic acid, glycine, asparagine, proline, serine, threonine, valine, alpha-amino-butyric acid, and norvaline; AA e and A.A. g is independently selected from a charged amino acid, including but not limited to aspartic acid, glutamic acid, lysine, arginine, ornithine, sulfo-serine, and phosphoserine; e is an integer selected from 1 to 6.

[0455] In some embodiments, AA a and A.A. dEach occurrence of is independently selected from leucine, isoleucine, and norleucine.

[0456] In some embodiments, AA b , A.A. c and A.A. f Each occurrence of is independently selected from alanine, proline, and serine.

[0457] In some embodiments, each occurrence of AAe and AAg is independently selected from aspartic acid, glutamic acid, lysine, arginine, and ornithine.

[0458] In some embodiments, each of E1 and / or E2 is independently (IAALESK) e , (IAALKSK) e , (IAALESE) e , (IAALKSE) e , (VAALKAE) e , (IAALKAE) e , (LAALKAE) e , (VSALKAE) e , (ISALKAE) e , (LSALKAE) e , (VASLKAE) e , (IASLKAE) e , (LASLKAE) e , (VSSLKAE) e , (ISSLKAE) e , (LSSLKAE) e , (VAALKSE) e , (LAALKSE) e , (VSALKSE) e , (ISALKSE) e , (LSALKSE) e , (VASLKSE) e, and (IASLKSE) e The heptad repeats are selected from:

[0459] In some embodiments, each of E1 and / or E2 is independently (KSELAAI) e , (KSKLAAI) e , (ESSLAAI) e , (ESKLAAI) e , (EAKLAAV) e , (EAKLAAI) e , (EAKLAAL) e , (EAKLASV) e , (EAKLASI) e , (EAKLASL) e , (EAKLSAV) e , (EAKLSAI) e , (EAKLSAL) e , (EAKLSSV) e , (EAKLSSI) e , (EAKLSSL) e , (ESKLAAV) e , (ESKLAAI) e , (ESKLAAL) e , (ESKLASV) e , (ESKLASI) e , (ESKLASL) e , (ESKLSAV) e , (ESKLSAI) e The heptad repeats are selected from:

[0460] In some embodiments, e is an integer selected from 1 to 4. In other embodiments, e is an integer selected from 2 or 3.

[0461] In some embodiments, 6 amino acids of each heptad are D amino acids, hi other some embodiments, 7 amino acids of each heptad are D amino acids.

[0462] In some embodiments, at least one of E1 and E2 is present in at least one peptide-antigen conjugate.

[0463] In some embodiments, each of E1 and / or E2 independently represents a group of formula (AA a -AA b -AA c -AA d -AA e -AA f -AA g ) e wherein: AA a and A.A. d is independently selected from leucine, isoleucine, norleucine, valine, norvaline, T-leucine, alloisoleucine, N-propylglycine, methionine, and O-methylserine; AA b , A.A. c and A.A. f is independently selected from alanine, cysteine, aspartic acid, glycine, asparagine, proline, serine, threonine, valine, alpha-amino-butyric acid, and norvaline; each occurrence of AAe and AAg is independently selected from a charged amino acid, including, but not limited to, aspartic acid, glutamic acid, lysine, arginine, ornithine, sulfo-serine, and phosphoserine; e is an integer selected from 1 to 6.

[0464] In some embodiments, AA a and A.A. d Each occurrence of is independently selected from leucine, isoleucine, and norleucine.

[0465] In some embodiments, AA b , A.A.c and A.A. f Each occurrence of is independently selected from alanine, proline, and serine.

[0466] In some embodiments, each occurrence of AAe and AAg is independently selected from aspartic acid, glutamic acid, lysine, arginine, and ornithine.

[0467] In some embodiments, each of E1 and / or E2 is independently (IAALESK) e , (IAALKSK) e , (IAALESE) e , (IAALKSE) e , (VAALKAE) e , (IAALKAE) e , (LAALKAE) e , (VSALKAE) e , (ISALKAE) e , (LSALKAE) e , (VASLKAE) e , (IASLKAE) e , (LASLKAE) e , (VSSLKAE) e , (ISSLKAE) e , (LSSLKAE) e , (VAALKSE) e , (LAALKSE) e , (VSALKSE) e , (ISALKSE) e , (LSALKSE) e , (VASLKSE) e , and (IASLKSE) e The heptad repeats are selected from:

[0468] In some embodiments, each of E1 and / or E2 is independently (KSELAAI) e , (KSKLAAI) e , (ESSLAAI) e , (ESKLAAI) e , (EAKLAAV) e , (EAKLAAI) e , (EAKLAAL) e , (EAKLASV) e , (EAKLASI) e , (EAKLASL) e , (EAKLSAV) e , (EAKLSAI) e , (EAKLSAL) e , (EAKLSSV) e , (EAKLSSI) e , (EAKLSSL) e , (ESKLAAV) e , (ESKLAAI) e , (ESKLAAL) e , (ESKLASV) e , (ESKLASI) e , (ESKLASL) e , (ESKLSAV) e , (ESKLSAI) e The heptad repeats are selected from:

[0469] In some embodiments, e is an integer selected from 1 to 4. In other embodiments, e is an integer selected from 2 or 3.

[0470] In some embodiments, 6 amino acids of each heptad are D amino acids, hi other some embodiments, 7 amino acids of each heptad are D amino acids.

[0471] The present disclosure also relates to a method of inducing an immune response in a subject in need thereof, comprising administering to the subject at least one dose of a first vaccine (V1), followed by at least one dose of a second vaccine (V2), where V1 is a vaccine disclosed herein and V2 is a viral vaccine.

[0472] 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).

[0473] 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).

[0474] In some embodiments, one dose of V1 is administered at a first time (V1T1). In some other embodiments, two doses of V1 are administered at a first time (V1T1) and a second time (V1T2). In some other embodiments, three doses of V1 are administered at a first time (V1T1), a second time (V1T2), and a third time (V1T3).

[0475] In some embodiments, one dose of V2 is administered at a first time (V2T1). In some other embodiments, two doses of V2 are administered at a first time (V2T1) and a second time (V2T2). In some other embodiments, three doses of V2 are administered at a first time (V2T1), a second time (V2T2), and a third time (V2T3).

[0476] In some embodiments, V1 is administered by intramuscular or intravenous route.

[0477] In some embodiments, V2 is administered by an intravenous route.

[0478] In some embodiments, the first dose of V2 is administered 1-6 weeks after the last dose of V1. In other embodiments, the first dose of V2 is administered 1-12 weeks after the last dose of V1.

[0479] In some embodiments, V2 is an adenovirus vector vaccine.

[0480] In some embodiments, the adenovirus encodes a peptide antigen (A) of V1.

[0481] In some embodiments, V2 is a ChAdOx vaccine. Linker

[0482] The term linker refers to any molecule that connects any two or more molecules (or "moieties") together, such as any two or more components of an amphiphile, peptide antigen conjugate, hapten conjugate, or drug conjugate, and may further perform any one or more of the following functions: I) increase or decrease aqueous solubility, II) increase the distance between any two components, III) affect rigidity or flexibility, or IV) modulate the degradation rate of the linkage between any two or more different molecules. As used herein, the term "linker" can be used to describe a linker (U), a suitable linker (X), such as X1, X2, X3, X4, and X5, and an extension (E1 or E2).

[0483] Particularly useful linkers are named, and specific preferred compositions of the named linkers are described throughout this specification. Thus, extensions E1 and E2 are optional peptide-based linkers extending from the N-terminus and C-terminus of the peptide antigen (A), respectively, and may be included between the solubilizing block (S) 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 to the solubilizing block or drug (D) or indirectly via the spacer (B), with linker precursor 2 ("U2") on the hydrophobic block (H), is referred to as linker U. A suitable linker X refers to any linker suitable for linking two or more adjacent groups. A suitable linker that is preferred for attaching a drug molecule (D) to the hydrophobic block (H) is referred to as X1. A suitable linker that is preferred for attaching an aryl or heteroaryl group to the hydrophobic block is referred to as X2. A suitable linker that is used to attach a reactive functional group ("FG4") to the pharmacophore of a drug molecule (D) is referred to as X3. A suitable linker that is preferred for attaching a charged group to the hydrophobic block (H) is referred to as X4. A suitable linker that is preferred for attaching SG to S is referred to as X5.

[0484] The linker can use covalent or non-covalent means to link any two or more components. In a preferred embodiment, the linker can link, i.e., connect, any two components by a covalent bond. A covalent bond is the preferred link used to link any two components, and the covalent bond ensures that neither component is readily dispersible from the other component after administration to a subject.

[0485] There are many suitable linkers well 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 for covalent linkers further comprise two or more functional groups, which may be the same or different, used to covalently link any two molecules, e.g., any two components of an amphiphile, a peptide antigen conjugate, and / or a drug conjugate.

[0486] In some embodiments, the carbon linker can include a C1-C18 alkane linker, e.g., a lower alkyl linker, e.g., C1-C6 (i.e., 1-6 methylene units), which can serve 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, can be used in place of an alkane linker to increase the spacing between any two or more heterogeneous molecules and to increase aqueous solubility. In other embodiments, the linker can be a cyclic and / or aromatic or 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).

[0487] For linking two components of amphiphiles, peptide-antigen conjugates, and drug conjugates, at least one of which comprises a peptide, linkers containing two to seven methylene groups have been found to improve coupling of two or more components. As 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) has improved coupling to other molecules, including U2, antigen (A), extension E2, spacer (B), and solubilizing block (S). Thus, in preferred embodiments, the N-terminal amino acid of the poly(amino acid)-based hydrophobic block (H) contains two or more, typically 2 to 7, 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.

[0488] In some embodiments, the linker can 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 any 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, can be 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 monomers in length, or longer. When used as a spacer (B), PEG can be up to 45 monomers in length, or longer, but is typically 4 to 36 monomers in length.

[0489] In some embodiments where the linker comprises a carbon chain, the linker can comprise a chain of about 1 or 2 to about 18 carbons in length, e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18 carbons, or more. In some embodiments where the linker comprises a carbon chain, the linker can comprise a chain of about 12 to about 20 carbons. In some embodiments where the linker comprises a carbon chain, the linker can comprise a chain of 18 carbons or less, typically about 1 to 6 carbon atoms.

[0490] The linkages used to join any two or more molecules, e.g., any two or more components of an amphiphile, a peptide antigen conjugate and / or a 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.

[0491] In a non-limiting example of covalent linkage, a click chemistry reaction can be used to obtain a triazole that links, i.e., bonds, any two components of an amphiphile, a peptide antigen conjugate, or a drug molecule conjugate to each other. In some embodiments, the click chemistry reaction is a strain-promoted [3+2] azide-alkyne cycloaddition reaction. An alkyne group and an azide group can be provided on each molecule and linked via "click chemistry." In some embodiments, an antigen (A) bearing an azide functional group is coupled with a hydrophobic block (H) bearing a suitable reactive group, such as an alkyne, e.g., dibenzylcyclooctyne (DBCO).

[0492] In some embodiments, amines may be provided on one molecule and linked to another molecule by reacting with any suitable electrophilic group, for example, a carboxylic acid, an acid chloride, an activated ester (e.g., an NHS ester), resulting in an amide bond; amines can be reacted with alkenes (via Michael addition); amines can be reacted with aldehydes and ketones (via Schiff bases); or amines can be reacted with activated carbonates or carbamates to give carbamates.

[0493] In some embodiments, the linker is cleavable under intracellular conditions, such that cleavage of the linker results in release of any moiety, e.g., a drug molecule (D), attached to the linker.

[0494] For example, the linker can be cleavable by an enzyme localized in an intracellular vesicle (e.g., in a lysosome, endosome, or caveolae) or by an enzyme in the cytosol, e.g., a proteasome or immunoproteasome. The linker can be, for example, a peptide linker that is cleaved by a protease enzyme, including, but not limited to, a protease localized in an intracellular vesicle, e.g., a cathepsin in the lysosomal or endosomal compartment of a cell.

[0495] Peptide linkers are typically 1 to 10 amino acids long, e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acids long, or longer (e.g., up to 20), e.g., 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 amino acids long, or longer. When used as a spacer (B), the peptide linker can be up to about 45 amino acids long. It is known that certain dipeptides are hydrolyzed by proteases, including cathepsins, e.g., cathepsins 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 (e.g., a Phe-Leu or Gly-Phe-Leu-Gly (SEQ ID NO: 1) linker) can be used. 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 examples of amino acids or peptides provided throughout this specification (including in the text of figures), unless otherwise specified, the peptides and amino acids should be understood to be L-amino acids.

[0496] 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.

[0497] In some embodiments, linkers comprising peptide sequences of the formula Pn...P4-P3-P2-P1 are used to facilitate recognition by cathepsins, 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 to the cleavage site C-terminal to the P1 residue, e.g., the amide bond between P1 and P1' is hydrolyzed. Suitable peptide sequences that promote cleavage by endosomal and lysosomal proteases, such as cathepsins, are well described in the literature (see Choe et al., J. Biol. Chem., 281:12824-12832, 2006).

[0498] 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, such as 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 that is 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 a cathepsin-cleavable linker. In some embodiments, the amino acid that promotes immunoproteasome processing is linked to a linker that promotes processing by endosomal proteases. Many suitable sequences for promoting immunoproteasome cleavage have been well described in the literature (see, for example, 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).

[0499] 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 shown here: [ka] where D is a drug molecule, "linker" is any suitable linker molecule, j is 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, either directly or indirectly through a terminal end, to the N- or C-terminus of a hydrophobic block (H) comprising, for example, a poly(amino acid), either directly or through a U, or by a reactive monomer that makes up the hydrophobic block (H), and the square brackets "[ ]" indicate that the group is optional.

[0500] 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]

[0501] In a non-limiting example of the above structure, if the N-terminal linker group is present and selected from beta-alanine, the structure becomes: [ka] is.

[0502] In some embodiments, the drug molecule (D) is linked to the peptide via a self-immolative carbamate linker. Non-limiting examples are shown here: [ka]

[0503] In the above example, if j is 4 and the amino acids are serine-lysine (Ac)-valine-norleucine, the structure is [ka] is.

[0504] In some embodiments, the drug molecule (D) is linked to the hydrophobic block (H) by a sulfatase-degradable linker X1, and the drug molecule is released from the linker as a result of hydrolysis of the sulfate by sulfatase. Several arylsulfatase- and alkylsulfatase-degradable linkers have recently been described (see, for example, 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) by a sulfatase-degradable linker. Non-limiting examples are provided here for clarity: [ka] wherein D is a drug molecule, "linker" is any suitable linker molecule linked, either directly or via a terminal, to the N-terminus or C-terminus of a hydrophobic block (H) comprising, for example, a poly(amino acid), either directly or via a U, or by a reactive monomer that makes up the hydrophobic block (H), and the square brackets "[ ]" indicate that the group is optional.

[0505] A non-limiting example of the above structure, where the "linker" is present and is selected from a short alkyl linker connected to the hydrophobic block by an amide, is shown here for clarity: [ka]

[0506] In other embodiments, any two or more components can be linked together by a pH-sensitive linker that is sensitive to hydrolysis under acidic conditions.Many pH-sensitive linkers are well known to those skilled in the art, including, for example, hydrazone, carbohydrazone, semicarbazone, thiosemicarbazone, cis-aconitic amide, orthoester, acetal, ketal, silyl ether or the like (see, for example, U.S. Patent 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).

[0507] In certain embodiments, different components (e.g., a drug molecule and a hydrophobic block (H)) are linked to each other by a pH-sensitive linker that is stable at blood pH, e.g., about pH 7.4, but hydrolyzes more rapidly at endosomal / lysosomal pH, e.g., about pH 5-6.5. In certain preferred embodiments, a drug molecule (D) is linked to a hydrophobic block (H) via a reactive monomer by a pH-sensitive bond, such as a hydrazone bond resulting from the reaction between a ketone and a hydrazine. While the functional group hydrazine linked to a carbonyl is sometimes referred to as a hydrazide, the term hydrazine is intended to refer broadly to the -NH-NH group, including when linked to a carbonyl, e.g., C(O)-NH-NH. A pH-sensitive linkage such as a hydrazone offers the advantage that the linkage is stable at physiological pH, e.g., about pH 7.4, but hydrolyzes at lower pH values, such as the pH of intracellular vesicles.

[0508] In certain preferred embodiments, the drug molecules are linked by a ketone-containing linker X1 and have the formula: [ka] (wherein D is any drug molecule, "linker" is any suitable linker molecule, y1 is an integer from 1 to 6, preferably 4, and square brackets "[ ]" indicate that the group is optional.) where the ketone in the above example is used to link the linker-linked drug molecule (D) to the reactive monomer via a hydrazone bond.

[0509] In the above example, if y1 is 4 and the drug molecules are directly linked by an amide bond (i.e., there is no "linker"), the structure would be: [ka] is.

[0510] In a preferred embodiment, the drug molecule linked to the ketone is linked to the hydrophobic block (H) by a hydrazone or carbohydrazone bond. Non-limiting examples of drug molecules linked to glutamic acid-based reactive monomers (N) by hydrazone and carbohydrazone bonds are shown here: [ka]

[0511] In some embodiments, the drug molecule may comprise a ketone and be directly linked to the reactive monomer by a hydrazone or carbohydrazone.

[0512] In other embodiments, the linker comprises a linkage that can be cleaved under reducing conditions, such as a reducible disulfide bond.Many different linkers that can be used to introduce disulfide bonds are known in the art (see, for example, Thorpe et al., 1987, Cancer Res. 47:5924-5931; Wawrzynczak et al., In Immunoconjugates: Antibody Conjugates in Radioimagery and Therapy of Cancer (CW Vogel, ed., Oxford U. Press, 1987); Phillips et al., Cancer Res. 68:9280-9290, 2008).See also U.S. Patent No. 4,880,935.

[0513] 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) includes an enzyme-degradable peptide and a self-immolative linker.

[0514] Although X can be any suitable linker, in preferred embodiments, the linker X linking any two or more groups is a short alkyl (i.e., lower alkyl) or PEG linker, e.g., a PEG linker having from about 1 to about 24 monomer units.

[0515] Extensions (E1 and E2): The optional N- and C-terminal extensions (E1 and E2) refer to moieties linked to the N- and C-termini, respectively, of the peptidic antigen (A). The N- and C-terminal extensions E1 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; or the like; or combinations thereof. The N- and C-terminal extensions E1 and E2 are attached to the peptidic antigen (A) by any suitable means, for example, by an amide bond.

[0516] In some embodiments, the extensions (E1 and E2) function to control the degradation rate of the peptidic antigen (A), but can also serve any one or more additional functions. In some embodiments, the N- or C-terminal extensions (E1 or E2) are free (in which case 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 can function to slow the degradation of the peptidic antigen. For example, an E1 peptide-based extension can 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) can be linked to heterologous molecules, and these extensions can also function as linkers to regulate the degradation of the peptidic antigen (A). The N- and / or C-terminal extensions providing the linker function can link the peptidic antigen to the hydrophobic block (H) or the soluble block (S) either directly or indirectly via a linker U. In some embodiments, the extensions (E1 and / or E2) function to provide distance, i.e., spacing, between any two heterologous molecules. In other embodiments, the extensions (E1 and / or E2) function to impart hydrophobic or hydrophilic properties to the peptide-antigen conjugate. In still other embodiments, the composition of the extensions (E1 and / or E2) can be selected to impart rigidity or flexibility. In other embodiments, the N-terminal and / or C-terminal extensions (E1 and / or E2) can serve to stabilize particles formed by the peptide-antigen conjugate.

[0517] In some embodiments, the extensions (El and / or E2) comprise 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 adjust the net charge of the peptide-antigen conjugate. Described below are peptide-based extensions (El and / or E2) that are recognized by proteases and impart specific electrostatic charges that stabilize particles formed by the peptide-antigen conjugates.

[0518] Additionally, in some embodiments, the C-terminal extension (E2) added to the peptide antigen (A) is selected to facilitate production of a peptide comprising the formula [S]-[E1]-A-E2-[U1] (where [ ] indicates that the group is optional). The sequence of the peptide-based E2 can be selected to prevent peptide beta-sheet formation and prevent sequence shortening during solid-phase peptide synthesis. In a non-limiting example, the C-terminal dipeptide linker (E2), Gly-Ser, is incorporated as a pseudoproline dipeptide (e.g., Gly-Ser(Psi(Me,Me)pro)) during solid-phase peptide synthesis. In a further embodiment, proline is incorporated into E2, e.g., Ser-Pro-Leu-Arg (SEQ ID NO: 4), thereby incorporating a proline to both facilitate production and promote processing of the extension by endosomal proteases.

[0519] In some embodiments, the peptide antigen (A) is linked at its C-terminus to an E2 extension, which is linked to a hydrophobic block either directly or indirectly via a linker (U), e.g., in this case the peptide antigen conjugate has the structure A-E2-UH or A-E2-H. In some embodiments, the E1 extension is linked to the N-terminus of the peptide antigen (A), and the E2 extension is linked to the C-terminus of the peptide antigen (A), and either E1 or E2 is linked to a hydrophobic block (H), either directly or via a linker (U), e.g., in this case 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, which is linked to the hydrophobic block (H) either directly or through a linker (U), e.g., in this case the peptide antigen conjugate has the structure HU-E1-A or H-E1-A. In some embodiments, the solubility block is linked to an extension, E1 or E2, which is linked to the N-terminus or C-terminus of the peptide antigen (A), respectively, and the extension not linked to the solubility block (S) is linked to the hydrophobic block (H) either directly or through a linker (U), e.g., in this case the peptide antigen conjugate has the structure S-E1-A-E2-UH, HU-E1-A-E2-S, E1-A-E2-H, or H-E1-A-E2-S.

[0520] In further embodiments, the soluble block (S) is linked to both extensions E1 and E2, which are linked to both the N-terminus and C-terminus of the peptidic antigen (A), respectively; or the soluble block (S) is linked to an E1 extension that is linked to the N-terminus of the peptidic antigen (A), but not to an E2 extension that is attached to the C-terminus of the peptidic antigen (A), which may be linked either directly to the hydrophobic block (H) or by a linker (U). The linker precursor U1 or linker (U) can be linked to either of the extensions (E1 or E2) by any suitable means, such as an amide bond.

[0521] 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.

[0522] 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 in length, that is linked to the peptide antigen (A) by an amide bond formed between the carboxyl group of E1 and the alpha amine of the N-terminal residue of the peptide antigen (A). The amide bond between E1 and the peptide antigen (A) can be enzymatically cleaved.

[0523] It is conventional to number amino acid positions C-terminal to the cleavage site (e.g., Pn'), indicated by a dash. For example, for a tetrapeptide extension (PN4-PN3-PN2-PN1), e.g., PN4-PN3-PN2-PN1-PA1'-PA2'-PA3'-PA4'-PA5'-PA6'-PA7'-PA8', linked to the N-terminus of a peptide antigen (A), which is an octapeptide (PA1'-PA2'-PA3'-PA4'-PA5'-PA6'-PA7'-PA8'), the amide bond between PN1 and PA1' is recognized and hydrolyzed by the enzyme.

[0524] 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, wherein 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.

[0525] In another embodiment, the N-terminal extension (E1) is an enzymatically degradable peptide recognized by the immunoproteasome, in which 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.

[0526] In a further embodiment, the N-terminal extension (El) is an enzymatically degradable peptide recognized by both endosomal proteases and immunoproteasomes, wherein the PN5 and PN1 positions of the octapeptide extension (PN8-PN7-PN6-PN5-PN4-PN3-PN2-PN1) are selected from arginine, lysine, citrulline, glutamine, threonine, leucine, norleucine, or methionine for the PN5 position recognized by cathepsins, and isoleucine, leucine, norleucine, or valine for the PN1 position recognized by immunoproteasomes, 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).

[0527] 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.

[0528] In some embodiments, E2 is a degradable peptide linked to the C-terminal residue of the peptide antigen (A) and includes 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 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. In a preferred embodiment, the C-terminal extension (E2) is linked to the peptide antigen (A) by 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 enzymatically cleaved. Note that for amino acid positions C-terminal to the cleavage site (e.g., Pn') indicated by a dash, it is conventional to number the amino acid positions from proximal to distal to the cleavage site. For example, for a tetrapeptide extension (PC1'-PC2'-PC3'-PC4'), e.g., PA8-PA7-PA6-PA5-PA4-PA3-PA2-PA1-PC1'-PC2'-PC3'-PC4', linked to the C-terminus of an octapeptide antigen (PA8-PA7-PA6-PA5-PA4-PA3-PA2-PA1), the amide bond between PA1 and PC1' is recognized and hydrolyzed by the enzyme.

[0529] 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, such as leucine, that promotes immunoproteasome hydrolysis of 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 of the amide bond proximal to the C-terminus of peptide antigen (A). Immunoproteasomes prefer small, uncharged amino acids at the PC1' position adjacent to the C-terminal amino acid PA1 of peptide antigen (A), for example, at 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, the C-terminal extension can be selected to promote recognition by either or both classes of proteases.

[0530] 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, the C-terminal extension E2 is a dipeptide PC1'-PC2', where PC1' is selected from Gly, Ala or Ser and PC2' is selected from Gly, Ala, Ser, Pro, Arg, Lys, Cit, Gln, Thr, Leu, Nle or Met. In a further embodiment, the C-terminal extension E2 is a tripeptide PC1'-PC2'-PC3', where 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.

[0531] In a further embodiment, the C-terminal extension E2 is a tetrapeptide extension PC1'-PC2'-PC3'-PC4' (wherein PC1' is selected from glycine, alanine or serine, PC2' is selected from glycine, alanine, serine, proline or leucine, PC3' is selected from glycine, alanine, serine, valine, leucine or isoleucine, and PC4' is selected from arginine, lysine, citrulline, glutamine, threonine, leucine, norleucine or methionine). In a further embodiment, the C-terminal extension, E2, is a pentapeptide PC1'-PC2'-PC3'-PC4'-PC5', where 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 a hexapeptide PC1'-PC2'-PC3'-PC4'-PC5'-PC6', where 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.

[0532] 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).

[0533] 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).

[0534] 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).

[0535] 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.

[0536] 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 immunoproteasomes and endosomal proteases. In a non-limiting example, a peptide antigen (A) having the sequence PA8-PA7-PA6-PA5-PA4-PA3-PA2-PA1 is linked to the C-terminus of 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), such as Ser-P3-P2-Arg. In some embodiments, an antigen having the sequence PA8-PA7-PA6-PA5-PA4-PA3-PA2-PA1 is linked to the C-terminal side of a C-terminal hexapeptide extension (E2) having the sequence PC1'-PC2'-PC3'-PC4'-PC5'-PC6' (wherein PC1' and PC2' are selected from glycine, alanine, proline, or serine, and PC6' is selected from arginine, lysine, citrulline, glutamine, threonine, leucine, norleucine, or methionine), e.g., Gly-Gly-PC3'-PC4'-PC5'-Arg. A non-limiting example of a C-terminal extension (E2) linked to the C-terminus of peptide antigen (A) that promotes processing by both immunoproteasomes and cathepsins is Gly-Gly-Lys-Pro-Leu-Arg (SEQ ID NO: 18). Further non-limiting examples of C-terminal extensions (E2) linked to the C-terminus of peptide antigens (A) that favor processing by the immunoproteasome and cathepsins are Gly-Gly-Ser-Leu-Val-Cit (SEQ ID NO: 20) or Gly-Gly-Ser-Pro-Val-Cit (SEQ ID NO: 21).

[0537] In some embodiments of the vaccine, the extensions (E1 and / or E2) have the formula (AA H -AA P -AA P -AA H -AA P -AA P -AA P )e wherein AA H is usually a hydrophobic amino acid suitable for coil domains, and AA P is typically a hydrophilic or small amino acid suitable for a coil domain, and e typically represents an integer selected from 1 to 6, for example, 1, 2, 3, 4, 5 and 6, preferably 1 to 4, most preferably 2 or 3. H -AA P -AA P -AA H -AA P -AA P -AA P ) e Each amino acid in the heptad repeat of should be carefully selected to ensure that the resulting sequence forms a helical secondary structure capable of forming a coiled-coil superstructure. To help guide the selection, H -AA P -AA P -AA H -AA P -AA P -AA P ) e The heptad repeat of can be more precisely described according to the following formula: (AA a -AA b -AA c -AA d -AA e -AA f -AA g ) e , in this formula, AA a and A.A. d is usually selected from hydrophobic amino acids, and AA b , A.A. c and A.A. f is usually selected from small amino acids, AA e and A.A. g are usually selected from charged amino acids. For clarity, amino acids refers to both proteinogenic and non-proteinogenic amino acids.

[0538] For purposes of selecting amino acids for use in the heptad repeats described herein, the general properties of proteinogenic amino acids, also called natural amino acids, as well as non-limiting examples of non-proteinogenic amino acids, also known as unnatural amino acids, are summarized in the table below. [Table A-1] [Table A-2] [Table A-3] [Table A-4]

[0539] The amino acid names are provided in the first column with parenthesized single-letter and, where appropriate, three-letter abbreviations. The physicochemical properties of the amino acids ("small," "aliphatic," "aromatic," "hydrophobic," "charged") are indicated as T ("true") or F ("false") and are provided as a guide for selecting amino acids for use in the heptad repeats described herein. The notation "charge" refers to the state of the amino acid at physiological pH, pH 7.4. For example, aspartic acid (D, Asp) is indicated as a small amino acid that is charged, e.g., charged at physiological pH (pH 7.4).

[0540] Formula (AA a -AA b -AA c -AA d -AA e -AA f -AA g ) e In certain embodiments of the heptad repeat of a and A.A. dEach occurrence of is independently selected from hydrophobic amino acids, more preferably aliphatic amino acids, including, but not limited to, leucine, isoleucine, norleucine, valine, norvaline, T-leucine, alloisoleucine, and N-alkylated amino acids, such as N-propylglycine, methionine, and alkylated alcohols, such as O-methylserine. a -AA b -AA c -AA d -AA e -AA f -AA g ) e In a preferred embodiment of the heptad repeat of a and A.A. d Each occurrence of is independently selected from leucine, isoleucine, and norleucine.

[0541] Formula (AA a -AA b -AA c -AA d -AA e -AA f -AA g ) e In certain embodiments of the heptad repeat of b , A.A. c and A.A. f Each occurrence of is independently selected from small amino acids, including, but not limited to, alanine, cysteine, aspartic acid, glycine, asparagine, proline, serine, threonine, valine, alpha-amino-butyric acid, and norvaline. a -AA b -AA c -AA d -AA e -AA f -AA g ) e In a preferred embodiment of the heptad repeat of b , A.A. c and A.A. f Each occurrence of is independently selected from alanine, proline, and serine.

[0542] Formula (AA a -AA b -AA c -AA d -AA e -AA f -AA g ) e In certain embodiments of the heptad repeat of e and A.A. g Each occurrence of is independently selected from a charged amino acid, including, but not limited to, aspartic acid, glutamic acid, lysine, arginine, ornithine, sulfo-serine, and phosphoserine. a -AA b -AA c -AA d -AA e -AA f -AA g ) e In a preferred embodiment of the heptad repeat of e and A.A. g is independently selected from aspartic acid, glutamic acid, lysine, arginine, and ornithine.

[0543] Formula (AA a -AA b -AA c -AA d -AA e -AA f -AA g ) e Non-limiting examples of heptad repeats include (IAALESK) e , (IAALKSK) e , (IAALESE) e , (IAALKSE) e , (VAALKAE) e , (IAALKAE) e , (LAALKAE) e , (VSALKAE) e , (ISALKAE) e , (LSALKAE) e, (VASLKAE) e , (IASLKAE) e , (LASLKAE) e , (VSSLKAE) e , (ISSLKAE) e , (LSSLKAE) e , (VAALKSE) e , (LAALKSE) e , (VSALKSE) e , (ISALKSE) e , (LSALKSE) e , (VASLKSE) e and (IASLKSE) e These include, but are not limited to:

[0544] For clarity, unless otherwise specified, the heptad repeats used in the extension sequences or linkers described herein may start and end at any position. For example, (AA a -AA b -AA c -AA d -AA e -AA f -AA g ) e (AA c -AA d -AA e -AA f -AA g -AA a -AA b ) e or (AA e -AA f -AA g -AA a -AA b -AA c -AA d ) e A non-limiting example is the heptad repeat (IAALESK) used in the extension (E1 and / or E2). eis a heptad repeat (ESKIAAL) e can be substituted with

[0545] Formula (AA a -AA b -AA c -AA d -AA e -AA f -AA g ) e AA of the heptad repeat a , A.A. b , A.A. c , A.A. d , A.A. e , A.A. f and A.A. g each occurrence of may be the same or different. e AA e Each occurrence of the formula (AA a -AA b -AA c -AA d -AA e -AA f -AA g ) e Non-limiting examples of heptad repeats are shown here for clarity: IAALKSKIAAKESE, IAALKSKIAAKESK, and IAALESEIAAKESK.

[0546] The unexpected discovery disclosed herein is that L-amino acids of the formula (AA a -AA b -AA c -AA d -AA e -AA f -AA g ) e The vaccine containing certain heptad repeats of the formula (AA) induced antibodies that bound to certain peptide sequences derived from endogenous proteins, including myosin proteins, but contained one or more D amino acids. a -AA b -AAc -AA d -AA e -AA f -AA g ) e Based on these unexpected findings, in certain preferred embodiments, a vaccine containing a particular heptad repeat of (AA a -AA b -AA c -AA d -AA e -AA f -AA g ) e The formula (AA H -AA P -AA P -AA H -AA P -AA P -AA P ) e The heptad repeat of formula (AA) contains one or more amino acids selected from D amino acids. a -AA b -AA c -AA d -AA e -AA f -AA g ) e Non-limiting examples of heptad repeats include (IAALESK) e , (IAALKSK) e , (IAALESE) e , (IAALKSE) e , (VAALKAE) e , (IAALKAE) e , (LAALKAE) e , (VSALKAE) e , (ISALKAE) e , (LSALKAE) e , (VASLKAE) e , (IASLKAE) e, (LASLKAE) e , (VSSLKAE) e , (ISSLKAE) e , (LSSLKAE) e , (VAALKSE) e , (LAALKSE) e , (VSALKSE) e , (ISALKSE) e , (LSALKSE) e , (VASLKSE) e , (IASLKSE) e where an italicized letter indicates that the amino acid is a D amino acid, and optionally up to 6 of the 7 amino acids in the heptad repeat can be L amino acids, e.g., 1 of the 7 amino acids in the heptad repeat is a D amino acid.

[0547] A further noteworthy finding is that the formula (AA H -AA P -AA P -AA H -AA P -AA P -AA P ) e , for example, containing L amino acids (AA) that induce antibodies that bind to certain peptide sequences derived from the myosin protein. a -AA b -AA c -AA d -AA e -AA f -AA g ) e For a particular heptad repeat in , the orientation of the heptad repeat can be determined by, for example, (AA a -AA b -AA c -AA d -AA e -AA f -AA g ) e From (AA g -AAf -AA e -AA d -AA c -AA b -AA a ) e By changing the heptad repeat from the heptad repeat sequence found in myosin to the heptad repeat sequence found in myosin, a non-limiting example of an alternative heptad repeat sequence to that found in myosin is (KSELAAI). e , (KSKLAAI) e , (ESSLAAI) e , (ESKLAAI) e , (EAKLAAV) e , (EAKLAAI) e , (EAKLAAL) e , (EAKLASV) e , (EAKLASI) e , (EAKLASL) e , (EAKLSAV) e , (EAKLSAI) e , (EAKLSAL) e , (EAKLSSV) e , (EAKLSSI) e , (EAKLSSL) e , (ESKLAAV) e , (ESKLAAI) e , (ESKLAAL) e , (ESKLASV) e , (ESKLASI) e , (ESKLASL) e , (ESKLSAV) e , (ESKLSAI) e These include, but are not limited to:

[0548] In some embodiments of the vaccine, one or more peptide antigen conjugates have the formula (AA H -AA P -AA P -AA H -AA P -AA P -AA P ) e , for example, (AA a -AA b -AA c -AA d -AA e -AA f -AA g ) e In a non-limiting example, the first peptide-antigen conjugate may each comprise an extension further comprising a heptad repeat of (IAALESK) e and the second peptide antigen conjugate may comprise an extension (IAALESK) e Or (IAALKSE) e or the first peptide antigen conjugate may comprise an extension (IAALESE) e and the second peptide antigen conjugate may comprise an extension (IAALKSK) e Or (IAALRSR) e Generally, the formula (AA a -AA b -AA c -AA d -AA e -AA f -AA g ) e For a vaccine comprising one or more peptide antigen conjugates having an extension comprising a heptad repeat of position AA e and A.A. g The amino acids in are selected to have a net charge of zero. For example, for a vaccine comprising a first conjugate and a second conjugate, the first conjugate may have the sequence (AA a -AA b -AAc -AA d -E-AA f -E) e For example, if the second conjugate contains an extension having (IAALESE), the second conjugate is selected to have a net charge of zero, typically (AA a -AA b -AA c -AA d -K-AA f -K) e , for example (IAALKSK), where K (lysine) can be optionally replaced with R (arginine) or ornithine.

[0549] For vaccines used to induce antibody responses, the linker molecule connecting the heptad repeat-containing extension to the peptide antigen has been found to affect the magnitude and quality of the antibody response. Thus, vaccines containing short and / or rigid amino acid linkers connecting the heptad repeat-based extension (E1 or E2) to the peptide antigen (A) resulted in higher functional antibody titers than vaccines containing longer and / or flexible linkers connecting the heptad repeat to the peptide antigen (A). A non-limiting explanation is that the rigid linker constrains the antigen, allowing for enhanced B cell receptor clustering compared to longer and / or flexible linkers. Thus, in certain preferred embodiments of vaccines comprising an extension (E1 or E2) selected from a heptad repeat, the heptad repeat is linked to either of the other components of the vaccine without a linker (i.e., the linker is absent), or the linker is selected from short and / or rigid linkers including, but not limited to, a single amino acid selected from, but not limited to, alanine, proline, β-alanine, N-ethyl-β-alanine, hydroxyproline, pipecolic acid, and stachydrine. Spacer (B)

[0550] 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., the amphiphile has the structure SBH or SBUH. The spacer (B) can comprise any one or more of the following: an amino acid, including an unnatural amino acid; a hydrophilic polymer, e.g., a polymer based on ethylene oxide (PEG), acrylate, methacrylate, acrylamide, or methacrylamide-based monomers; an alkane chain; or the like; or a combination thereof. The spacer (B) can be linked to the solubilizing block (S) and the hydrophilic block (H) by any suitable means, e.g., directly or indirectly via a linker, although the linkage typically comprises a covalent bond, e.g., an amide bond.

[0551] 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 an enzyme and facilitate degradation.

[0552] In some embodiments, the spacer (B) is a hydrophilic polymer having monomer units 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, phosphoesters, phosphonamides, sulfonate esters, sulfonamides, or combinations thereof.

[0553] In some embodiments, spacer (B) is a peptide sequence 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 in length, which is linked to hydrophobic block (H) and solubilizing block (S), respectively, by, e.g., amide bonds formed between the N-terminal and C-terminal carboxyl groups of spacer (B). 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 can be selected for resistance to enzyme-mediated hydrolysis.

[0554] 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, or HEMA, 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 or less in length, and linked to the hydrophobic block (H) and the solubilizing block (S) either directly or via a linker.

[0555] Specific spacer compositions that result in unexpected enhancements in biological activity are described throughout this specification. Note: While both the spacer group (B) and the solubilizing block (S) can 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.), the distinction between S and B is based in part on function, and caution is invoked in the specific instance of amphiphiles. Linker(U)

[0556] 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).

[0557] The linker (U) also binds the peptide antigen conjugate fragment ([S]-[E1]-A-[E2]-U1 or U1-[E1]-A-[E2]-[S]) to the hydrophobic block (H) fragment (U2-H) by reaction of U1 with U2 to form the peptide antigen conjugate ([S]-[E1]-A-[E2]-UH or HU-[E1]-A-[E2]-[S]).

[0558] Although the peptide antigen (A) can be coupled to the hydrophobic block (H) either directly, i.e., AH, or via an extension, i.e., A-E2-H (or H-E1-A), by solid-phase peptide synthesis entirely on resin, it is advantageous under certain circumstances to generate the antigen (A) and the hydrophobic block (H) as separate fragments comprising the linker precursor U1 ([S]-[E1]-A-[E2]-U1 or U1-[E1]-A-[E2]-[S]) and the linker precursor U2 (U2-H), which can be coupled on resin or in solution to give [S]-[E1]-A-[E2]-UH (or HU-[E1]-A-[E2]-[S]).

[0559] Similarly, the solubilizing block (S) on the amphiphile can be attached to the hydrophobic block (H) either directly, i.e., SH, or via a spacer, i.e., SBH, entirely on the resin by solid-phase peptide synthesis. However, under certain circumstances, it may be beneficial 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 can be attached on the resin or in solution to give S-[B]-UH.

[0560] 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 any other groups. In some embodiments, the linker precursor U1 comprises an activated carboxylic acid, which is reacted with an amine-containing linker precursor U2 to form an amide-containing linker U, or U1 comprises an amine, which 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, which is reacted with a thiol-containing linker precursor U2 to form a thioether-containing linker U, or U1 comprises a thiol, which is reacted with a maleimide-containing U2 to form a thioether-containing linker U. In some embodiments, the linker precursor U1 comprises an azide, which is reacted with an alkyne-containing linker precursor U2 to form a triazole-containing linker U, or U1 comprises an alkyne, which is reacted with an azide-containing U2 to form a triazole-containing linker US.

[0561] In a preferred embodiment, amphiphiles of formula S-[B]-UH are conjugated to one another 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), or the like), which is reacted with an azide-containing linker precursor U2 to form a triazole-containing linker U.

[0562] In a preferred embodiment, peptide antigen conjugates of the formula [S]-[E1]-A-[E2]-UH or HU-[E1]-A-[E2]-[S] are attached to one another by linking the peptide antigen fragment [S]-[E1]-A-[E2]-U1 or U1-[E1]-A-[E2]-[S] to a hydrophobic blocking fragment (U2-H), where the linker precursor U1 contains a strained alkyne (e.g., dibenzocyclooctyne (DBCO), bicyclononyne (BCN), or the like) which is reacted with an azide-containing linker precursor U2 to form a triazole-containing linker U.

[0563] In another preferred embodiment, the linker precursor U1 contains an azide, which is reacted with a strained alkyne (e.g., dibenzocyclooctyne (DBCO), bicyclononyne (BCN), or the like)-containing linker precursor U2 to form a triazole-containing linker U. In a non-limiting example, a linker precursor U2 comprising DBCO (e.g., DBCO-NHS, CAS No. 1353016-71-3) is linked to a hydrophobic block (H) via a suitable linker X, and a linker precursor U1 (e.g., an azido acid, e.g., azidopentanoic acid; an azidoamino acid, e.g., azido-lysine (abbreviated as Lys(N3), CAS No. 159610-92-1); or an amidoamine, e.g., azido-butylamine) is linked to a solubilizing block fragment (S-[B]-U1) or a peptide antigen fragment ([S]-[E1]-A-[E2]-U1 or U1-[E1]-A-[E2]-[S]) via a suitable linker X.

[0564] In a preferred embodiment, the linker U preferably comprises an amide, a thioether or a triazole. Dendron Amplifier

[0565] 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 the solubilizing block (S), the dendron amplifier is used to increase the valency of the solubilizing groups (referred to in the formula as "SG") present on the surface of the solubilizing block (S). In other embodiments, the dendron amplifier is used to increase the valency of the solubilizing block (S) and spacer (B) linked to the hydrophobic block (H).

[0566] Dendron amplifiers (also called "dendrons") are regularly branched, often symmetrical, molecules that typically contain repeating units of monomers 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 central and terminal functional groups, respectively, selected from any suitable functional group, 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 defined as FGt=β. γ where β is the number of branches occurring in each generation of the dendron and the symbol γ is the number of generations, where the number of branches can be any integer but is typically 2 to 6, and the number of generations can be any integer but is typically 1 to 10. The terminal functional groups present on the solubilizing block that are free (i.e., unreacted) are also referred to as solubilizing groups (SG).

[0567] A dendron amplifier can comprise repeats of a monomer comprising a first functional group (FG1) and a second functional group (FG2), where the first functional group is reactive to the second functional group. For example, a non-limiting example of a second generation dendron amplifier, where β=2, comprising repeats of a monomer comprising a first functional group (FG1) and a second functional group (FG2), where the first functional group is reactive to the second functional group, is shown here for clarity: [ka] Here, the first functional group at the starting point is also called the central functional group (FG') and the terminal FG2 is also called the terminal functional group or FGt.

[0568] A non-limiting example of a third generation dendron formed from a monomer containing a first and second functional group, where β=2, is shown here for clarity: [ka]

[0569] A non-limiting example of a second generation dendron amplifier, β=3, comprising repeats of a first monomer comprising a first functional group (FG1) and a second functional group (FG2), where the first functional group is reactive towards a second functional group, is shown here for clarity: [ka]

[0570] The monomer comprising a first functional group and a second functional group, wherein the first functional group is reactive to the second functional group, and 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, in which R 1 are independently (CH2) y3 -FG2, (OCH2CH2) y3 -FG2 or CH2(OCH2CH2) y3 -FG2), and y2 and y3 are each an integer number of repeating units selected from 1 to 6.

[0571] 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]

[0572] When the above monomers are used to generate a second generation amplification linker, the structure is [ka] is.

[0573] Further 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 monomers comprising at least one first functional group and two or more second functional groups, such as FG1-(CH2) y2 N(R 2 )2, FG1-(CH2CH2O) y2 CH2CH2N(R 2 ) 2, in which R 2 are independently (CH2) y3 -FG2, (CH2CH2O) y3 (CH2) y4 -FG2, (CH2OCH2CH2) y3-FG2), where y2, y3, and y4 are each integers selected from 1 to 6 of the repeating units. Note: In the above example, FG' is an amine and 4FGt is a carboxylic acid.

[0574] FG1-(CH2CH2O) y1 CH2CH2N(R 2 )2 (wherein FG1 is NH2 and R 2 is (CH2CH2O) y3 (CH2) y4 -FG2, y2 is 2, y3 is 1, y4 is 2, and FG2 is COOH) are shown here for clarity: [ka]

[0575] In yet a further non-limiting example of a monomer comprising a first functional group and a second functional group, wherein the first functional group is reactive to the second functional group, the monomer comprises at least one first functional group and two or more second functional groups comprise certain amino acids, e.g., glutamic acid, aspartic acid, lysine, or ornithine. A non-limiting example of a third generation lysine dendron is provided here for clarity. [ka]

[0576] A dendron amplifier can include two repeats of monomers, where the first monomer includes three or more first functional groups (FG1) and the second monomer includes 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 amplifier, where β=2, including repeats of first and second monomers, where the first monomer includes three first functional groups (FG1) and the second monomer includes two second functional groups (FG2), and the first functional groups are reactive to the second functional groups, is shown here for clarity: [ka]

[0577] A non-limiting example of a first generation dendron amplifier, β=2, comprising a repeat of first and second monomers, where the first monomer comprises three first functional groups (FG1), 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]

[0578] The dendron amplifiers can be used to link any three or more components of the amphiphile, peptide antigen conjugate, and drug molecule conjugate together. The central functional group (FG') and terminal functional groups (FGt) can be further functionalized, i.e., reacted, for a specific purpose.

[0579] In a preferred embodiment of an amphiphile of formula S-[B]-[U]-H, the solubilizing block (S) comprises a dendron amplifier whose core is linked either directly to the hydrophobic block (H) or indirectly via a spacer (B) and / or a linker U, and whose terminal functional groups (FGt) are either unlinked and serve as solubilizing groups, or linked to solubilizing groups (SG). The solubilizing groups (SG) are any hydrophilic and / or charged molecules, and preferred solubilizing groups (SG) are described elsewhere herein.

[0580] In some embodiments of amphiphiles of formula S-[B]-[U]-HD, peptide antigen conjugates of formula [S]-[E1]-A-[E2]-[U]-HD or HD-[U]-[E1]-A-[E2]-[S], and drug molecule conjugates of formula HD, the hydrophobic block (H) comprises a core (i) linked to the solubilizing block (S), either directly or indirectly through a spacer (B) and / or a linker U, (ii) linked to the antigen (A), either directly or indirectly through an extension (E1 or E2) and / or a linker U, or (iii) linked to the drug molecule, either directly or through a linker X1.

[0581] 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 core is (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. Capped or 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 can be capped with an activated carboxylic acid (e.g., acetyl chloride) to give a relatively unreactive amide, or, for example, a strained alkyne can be capped with an alkyl-azide to give a relatively unreactive triazole. Hydrophobic block (H)

[0582] The hydrophobic block (sometimes designated "H" in the formula) is a molecule with substantially limited aqueous solubility or is amphiphilic in nature and can assemble in aqueous solution to construct supramolecular structures, e.g., micelles, nanoparticles, or microparticles. In certain embodiments, the hydrophobic block (H) is insoluble in aqueous solution or forms micelles in aqueous solution at a concentration of about 1.0 mg / mL or less, 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 thermoresponsive hydrophobic polymer, 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, at physiological temperature (about 37° C.) and at or near physiological salt concentrations (about 10 g / L) and salt compositions.

[0583] The hydrophobic block (H) can be selected from any molecule having limited water solubility and / or amphiphilic properties, including higher alkanes, cyclic aromatics, fatty acids, terpene / isoprene-derived compounds, or polymers or oligomers.

[0584] Exemplary higher alkanes include, but are not limited to, octane, nonane, decane, undecane, dodecane, tridecane, tetradecane, pentadecane, hexadecane, heptadecane, and octadecane. Exemplary cyclic aromatics include, but are not limited to, phenyl. Exemplary saturated and unsaturated fatty acids include, but are not limited to, myristic acid, palmitic acid, stearic acid, or oleic acid. In some embodiments, the hydrophobic block (H) is a fatty acid, such as myristic acid. In other embodiments, the hydrophobic block (H) comprises a diacyl lipid, such as 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine or 1,2-distearoyl-sn-glycero-3-phosphoethanolamine, or a lipopeptide, such as Pam2Cys. In some embodiments, the fatty acid or lipid-based hydrophobic block (H) may further comprise PEG. Exemplary compounds derived from terpenes / isoprenes 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 saponin, such as QS-21.

[0585] 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 type or many different types of monomer units. The hydrophobic block (H) can be a statistical copolymer or an alternating polymer. The hydrophobic block (H) can be a block copolymer, such as an AB type, or the polymer can comprise a graft type copolymer, in which two or more polymers are linked by a polymer-analogous reaction.

[0586] The hydrophobic block (H) may comprise a polymer containing naturally occurring and / or non-naturally occurring monomers and combinations thereof.

[0587] In some embodiments, the hydrophobic block (H) is selected from natural biopolymers. Natural biopolymers can include peptides (sometimes referred to as poly(amino acids)) containing hydrophobic 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, for example, by acetylation or benzoylation, to have hydrophobic properties. Natural biopolymers that are water-soluble in their natural form can be used, but must be chemically modified to render them water-insoluble or suitable for use as the hydrophobic block (H). For example, biopolymers composed of hydrophilic amino acids, such as glutamic acid or lysine residues, can be modified with gamma carboxyl or epsilon amine groups, respectively, for attachment of hydrophobic molecules, such as hydrophobic drug molecules, to increase the hydrophobicity of the resulting modified biopolymer. Similarly, the biopolymer can be selected from hydrophilic polysaccharides, which may include, but are not limited to, glycogen, cellulose, dextran, alginate, and chitosan, but such polysaccharides should be chemically modified, for example, by acetylation or benzoylation of 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.

[0588] The monomer constituting the hydrophobic block (H) can 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, phosphoesters, phosphonamides, sulfonate esters, sulfonamides, or combinations thereof. Specific examples of (meth)acrylates and (meth)acrylamides include benzyl methacrylamide (BnMAM) and benzyl methacrylate (BnMA), respectively.

[0589] 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 solutions. Non-limiting examples include temperature-responsive monomers, such as N-isopropylmethacrylamide (NIPMAM); a homopolymer composed entirely of NIPMAM may be water-soluble at room temperature, but may become insoluble and form particles at elevated temperatures. This distinction is made to facilitate the description of certain embodiments. In some embodiments, the hydrophobic block (H) comprises a majority of monomer units selected from temperature-responsive hydrophobic monomers (sometimes referred to as "temperature-responsive monomers"), such as NIPAM, NIPMAM, N,N'-diethylacrylamide (DEAAM), N-(L)-(1-hydroxymethyl)propylmethacrylamide (HMPMAM), N,N'-dimethylaminoethyl methacrylate (DMEMA), N-(N-ethylcarbamido)propylmethacrylamide, N-vinylisobutyramide (PNVIBA), N-vinyl-n-butylamide (PNVBA), N-acryloyl-N-propylpiperazine (PNANPP), N-vinylcaprolactam (PVCa), DEGMA, TEGMA, or poly(amino acid) or γ-(2-methoxyethoxy)esteryl-L-glutamate. In yet other embodiments, the hydrophobic block (H) may comprise monomers of ethylene oxide, propylene oxide, or combinations thereof.

[0590] The hydrophobic block (H) comprising a polymer may typically comprise 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 a hydrophobic drug molecule. 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 other 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.

[0591] In a preferred embodiment, the hydrophobic block (H) comprises a polymer (or oligomer) containing 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 g...

Claims

1. an amphiphile having the formula S-[B]-[U]-H; at least one peptide antigen conjugate having a formula selected from: [S]-[E1]-A-[E2]-[U]-H and H-[U]-[E1]-A-[E2]-[S]; 1. A vaccine comprising: During the ceremony, S is, independently for each occurrence, a solubilizing block; B is a spacer, H, independently for each occurrence, is a hydrophobic block; one or more drug molecules (D) are optionally attached to each H, either directly or via a suitable linker, X1; A is, independently for each occurrence, a peptide antigen; E1, independently for each occurrence, is an N-terminal extension; E2, independently for each occurrence, is a C-terminal extension, U is, independently for each occurrence, a linker; [ ] indicates that the group is optional, - indicates that two adjacent groups are covalently linked to each other directly or indirectly via a suitable linker X; A vaccine, wherein the amphiphile and / or the at least one peptide antigen conjugate comprises a dendron amplifier.

2. The vaccine of claim 1 , wherein the amphiphile S comprises a dendron amplifier.

3. The vaccine of claim 1 or 2, wherein the dendron amplifier comprises 1 to 10 generations of repeating monomeric units with between 2 and 6 branches per generation.

4. The repeating monomer units are (a) 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 , FG1-CH(R １ ) ２ , and FG1-C(R １ ) ３ is selected from During the ceremony, R １ is independently for each occurrence, (CH ２ ) ｙ３ -FG2, (OCH ２ CH ２ ) ｙ３ -FG2, and CH ２ (O.C.H. ２ CH ２ ) ｙ３ -FG2), y2 and y3, independently for each occurrence, are an integer from 1 to 6 repeat units; FG1 is a first functional group, FG2 is a second functional group; or (b) selected from hydroxy acids, amino acids, polyols, polyamines, and amino alcohols; The vaccine of claim 3.

5. The vaccine of claim 3 or 4, wherein the dendron amplifier comprises a polyethylene oxide (PEG) group.

6. 6. The vaccine of any one of claims 1 to 5, 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.

7. Each H is independently selected from the formula: -(M) ｍ -(N) Ｎ -(O) ｏ -(P) ｐ -R ３ and a poly(amino acid) having the formula: each M is independently a hydrophobic amino acid; each N is independently a reactive amino acid; each O is independently a spacer amino acid; each P is independently a charged amino acid; m, n, o and p each independently represent an integer from 0 to 100, the sum of m, n, o and p is less than or equal to 100, and at least one of m and n is not 0; R ３ is a hydrogen, NH, connected either directly or by a suitable linker X ２ , NH-CH ３ , NH-(CH ２ ) ｙ５ CH ３ , OH or D; y5 is an integer from 1 to 6; A vaccine according to any one of claims 1 to 5.

8. P, if present, 【Chemistry 251】 wherein each R ５ is independently a group containing 1-2 charged functional groups; and / or When O is present, 【Chemistry 252】 wherein each Q is independently selected from (CH 2 ) y6 and (CH 2 CH 2 O) y7 CH 2 CH 2 , 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, and / or N, if present, 【Chemistry 253】 wherein each X1 is independently a suitable linker and each D is independently a drug molecule; and / or M, if present, 【Chemical 254】 wherein each R 4 is independently 【255】 wherein a is aryl or heteroaryl; X2 is present or absent and, if present, is a suitable linker; y8 is selected from an integer from 0 and 6; and Z 1 , Z 2 , and Z 3 are each independently selected from hydrogen, fluorine, hydroxy, amino, alkyl, and fluoroalkyl. The vaccine of claim 7.

9. Each R ４ But independently, 【Chemistry 260-1】 【Chemistry 260-2】 【Chemistry 260-3】 The vaccine of claim 8, which is selected from the group consisting of

10. At least one D is 【Chemical 263】 where: R ２０ is selected from hydrogen, alkyl, alkoxyalkyl, aryl, heteroaryl, aminoalkyl, amide and ester; X3 is selected from alkyl, alkoxyalkyl, aralkyl, heteroaralkyl, aryl, heteroaryl and carboxy; A vaccine according to any one of claims 1 to 9.

11. 11. The vaccine of any one of claims 1 to 10, wherein B, if present, is a hydrophilic polymer or peptide.

12. 12. The vaccine of any one of claims 1 to 11, wherein the U of the amphiphile and / or the U of the peptide antigen conjugate, if present, comprises an amide, a thioether or a triazole.

13. A vaccine described in any one of claims 1 to 12, wherein at least one A comprises a sequence in which one or more cysteine ​​residues are replaced with alpha amino-butyric acid and / or one or more methionine residues are replaced with norleucine.

14. 14. The vaccine of any one of claims 1 to 13 for treating or preventing cancer, an autoimmune disease, an allergy, an infectious disease, a cardiovascular disease or a neurodegenerative disease in a subject in need thereof.

15. The at least one peptide antigen conjugate is (a) selected from minimal immunogens; (b) selected from autoantigens, alloantigens and allergens; or (c) selected from tumor antigens; 14. The vaccine of any one of claims 1 to 13, comprising A.

16. 16. The vaccine of any one of claims 1 to 13 and 15, wherein E1 and E2 each comprise a PEG group of between 4 and 36 monomer units, or wherein E1 and E2 each comprise a peptide.

17. The vaccine comprising at least one D and, optionally, a second drug molecule (D2) and / or a third drug molecule (D2); said at least one D is each independently selected from (a) an inhibitor of mTOR, RORγt, CDK8 / 19 and HDAC and an agonist of AHR, RAR and A2a, or (b) an agonist of TLR-3, TLR-7, TLR-8, TLR-7 / 8, TLR-9 and STING; D2 is selected from an immunostimulant selected from an inhibitor of mTOR, RORγt, CDK8 / 19 and HDAC, and an agonist of AHR, RAR and A2a, and an agonist of NLR, CLR, TLR and STING; D3 is selected from an immunostimulant selected from an inhibitor of mTOR, RORγt, CDK8 / 19 and HDAC, and an agonist of AHR, RAR and A2a, and an agonist of NLR, CLR, TLR and STING; provided that D, D2 and D3 bind to different receptors; 17. A vaccine according to any one of claims 1 to 13 and 15 to 16.

18. A vaccine as described in any one of claims 1 to 14 and 15 to 17 for inducing tolerance in a subject in need thereof, wherein at least one A is selected from an autoantigen, an alloantigen and an allergen.

19. 20. A vaccine according to any one of claims 1 to 14 and 15 to 18 for the treatment or prevention of allergy in a subject in need thereof.

20. 19. The vaccine of any one of claims 1 to 14 and 15 to 18, wherein S is present for at least one peptide-antigen conjugate and comprises SG selected from an amine or a carboxylic acid.

21. 20. The vaccine of any one of claims 1 to 14 and 15 to 19, wherein the peptide-antigen conjugate has a net positive charge of between about +1 to about +10 at physiological pH or a net negative charge of between about -1 to about -10 at physiological pH.

22. 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 amino-butyric acid and / or one or more methionine residues are replaced with norleucine.

23. 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; and / or 22. The vaccine of claim 21, wherein at least one D is selected from an immunostimulant and a Treg-promoting immunomodulator.

24. 1. A vaccine comprising at least one peptide antigen conjugate having a formula selected from: [S]-[E1]-A-[E2]-[U]-H and H-[U]-[E1]-A-[E2]-[S], During the ceremony, S is, independently for each occurrence, a solubilizing block; H, independently for each occurrence, is a hydrophobic block; one or more drug molecules (D) are optionally attached to each H, either directly or via a suitable linker, X1; A is, independently for each occurrence, a peptide antigen; E1, independently for each occurrence, is an N-terminal extension; E2, independently for each occurrence, is a C-terminal extension, U is, independently for 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 and at least one D is present and selected from an agonist of TLR-7 / 8, and the vaccine further comprises a second drug molecule (D2) selected from an inhibitor of mTOR; (iii) at least one A is a glycopeptide; or (iv) at least one A is selected from an autoantigen, an allergen, and an alloantigen, and at least one D is present and selected from an ATP-competitive mTOR inhibitor. Either [ ] indicates that the group is optional, - indicates that two adjacent groups are covalently linked directly to each other or indirectly to each other via a suitable linker X.

25. A vaccine comprising an expression system comprising DNA or RNA encoding at least one peptide antigen (A), further comprising at least one drug molecule (D) selected from Treg-promoting immunomodulators.

26. A peptide antigen conjugate having a formula selected from S-[E1]-A-[E2]-[U]-H-[D] and [D]-H-[U]-[E1]-A-[E2]-S, or a peptide antigen fragment having a formula selected from S-[E1]-A-[E2]-[U1] and [U1]-[E1]-A-[E2]-S, During the ceremony, S is a solubilizing block; H is a hydrophobic block; one or more drug molecules (D) are optionally attached to each H, either 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, [ ] indicates that the group is optional, - indicates that two adjacent groups are covalently linked to each other directly or indirectly via a suitable linker X; S comprises one or more amino acids; Peptide-antigen conjugates or peptide-antigen fragments.

27. A vaccine for activating, priming and / or expanding T cells comprising the peptide antigen conjugate or peptide antigen fragment of claim 26.

28. 21. The vaccine according to any one of claims 1 to 14 and 15 to 20 for inducing an immune response in a subject in need thereof, wherein the vaccine is administered for at least one dose, followed by administration of a second vaccine (V2) for at least one dose, V2 is a viral vaccine; vaccine.

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