Compositions and uses of vasoactive intestinal peptide (VIP) antagonists

Abstract

The present disclosure relates to VIP antagonists for use in managing the treatment or prevention of cancer and viral infections. In certain embodiments, the present disclosure relates to chimeric variants of VIP antagonists as peptides disclosed herein and pharmaceutical compositions comprising the same. In certain embodiments, the present disclosure contemplates a method of targeting cancer by stimulating immune cells, combining the immune cells in vitro with the peptides disclosed herein, and administering an effective amount of the stimulated immune cells to a subject in need of cancer treatment.

Description

Detailed Description of the Invention

[0001] [Technical field] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Provisional Patent Application No. 62 / 768,060, filed November 15, 2018, the entirety of which is incorporated herein by reference for all purposes.

[0002] Incorporation by reference of material submitted as a text file via the Office Electronic Filing System (EFS-WEB) The sequence listing associated with this application is provided in text form in lieu of a paper copy and is incorporated herein by reference. The name of the text file containing the sequence listing is 18223PCT_ST25.txt. The text file is 12 KB, was created on November 15, 2019, and is submitted electronically via EFSWeb. [Background technology]

[0003] Vasoactive intestinal peptide (VIP) is produced by various cells including immune cells, neurons and endocrine cells in the central nervous system. Endogenous VIP is present in the nerves of airway smooth muscle and pulmonary blood vessels in the lungs, where VIP functions as a bronchodilator. VIP also has the ability to alter cell proliferation and the production of inflammatory signals through the VIP receptors VPAC1 and VPAC2. A chimeric peptide called VIPhyb was expressed with an N-terminal sequence that provides the C-terminal 22 amino acid sequence of VIP followed by membrane permeabilization. As the six N-terminal amino acids of native VIP were replaced, VIPhyb had altered biological activity acting as a VIP antagonist. VIP antagonists have also been reported in U.S. Patents 6,630,124 and 5,217,953.

[0004] Traditionally, cancer treatments usually use surgery, chemotherapy and radiation therapy. However, alternative methods have been reported to enhance the immune system to attack cancerous cells. These methods include harvesting, expanding and altering T cells to target and stimulate the immune system to aggressively eliminate cancerous cells. In chimeric antigen receptor (CAR) T cell therapy, isolated T cells are engineered to express a chimeric protein and are then infused back into the patient. However, there is a need to identify improved treatments.

[0005] Petersen et al. report that administration of VIPhyb enhances autologous anti-leukemic T cell responses in a mouse model of acute leukemia. Oncoimmunology, 2017, 6(5):e1304336. Petersen et al. report that administration of VIPhyb improves T cell proliferation and functions for adoptive T cell therapy using ex vivo treatment with a PI3Kδ inhibitor and a VIP antagonist. Blood Adv. 2018, 2(3):210-223.

[0006] The references cited herein are not admissions of prior art. [Summary of the Invention]

[0007] The present disclosure relates to VIP antagonists for use in managing the treatment or prevention of cancer and viral infection.In certain embodiments, the present disclosure relates to chimeric variants of VIP antagonists as peptides disclosed herein and pharmaceutical compositions comprising the variants.In certain embodiments, the present disclosure contemplates a method of targeting cancer by stimulating immune cells, mixing immune cells in vitro with peptides disclosed herein, and administering an effective amount of stimulated immune cells to a subject in need of cancer treatment.

[0008] In certain embodiments, the VIP antagonist is KPRRPYX. 1 X 2 NX 3 TX 4 LRKQX 5 AVX6 KYX 7 NX 8 A peptide containing ILN (SEQ ID NO: 11), 1 is A or any amino acid, and X 2 is V or any amino acid, and X 3 is C or any amino acid, and X 4 is S or any amino acid, and X 5 is I or any amino acid, and X 6 is N or any amino acid, and X 7 is M or any amino acid, and X 8 is I or any amino acid, and X is not a peptide of KPRRPYTDNYTRLRKQMAVKKYLNSILN (SEQ ID NO:1) or a combination. 1 is T and X 2 is D and X 3 is Y and X 4 is R and X 5 is M and X 6 is K and X 7 is L and X 8 is S.

[0009] In certain embodiments, the peptide is KPRRPYX 1 X 2 NX 3 TX 4 LRKQX 5 AVX 6 KYX 7 NX 8 ILN (SEQ ID NO: 11), 1 is A or T, and X 2 is V and X 3 is C and X 4 is S and X 5 is I and X 6 is N and X 7 is M, or X 8 is I if and only if X 1 is T;X 2 is V or D, and X 1 is A and X 3is C and X 4 is S and X 5 is I and X 6 is N and X 7 is M, or X 8 is I if and only if X 2 is D;X 3 is C or Y, and X 1 is A and X 2 is V and X 4 is S and X 5 is I and X 6 is N and X 7 is M, or X 8 is I if and only if X 3 is Y;X 4 is S or R, and X 1 is A and X 2 is V and X 3 is C and X 5 is I and X 6 is N and X 7 is M, or X 8 is I if and only if X 4 is R;X 5 is I or M, and X 1 is A and X 2 is V and X 3 is C and X 4 is S and X 6 is N and X 7 is M, or X 8 is I if and only if X 5 is M;X 6 is N or K, and X 1 is A and X 2 is V and X 3 is C and X 4 is S and X 5 is I and X 7 is M, or X 8 is I if and only if X 6 is K;X 7 is M or L, and X 1 is A and X 2 is V and X 3 is C and X 4 is S and X 5is I and X 6 is N, or X 8 is I if and only if X 7 is L;X 8 is I or S, and X 1 is A and X 2 is V and X 3 is C and X 4 is S and X 5 is I and X 6 is N, or X 7 is X if and only if M 8 is S.

[0010] In certain embodiments, the peptide is KPRRPYADNYTRLRKQMAVKKYLNSILN(SEQ ID NO:3), KPRRPYTVNYTRLRKQMAVKKYLNSILN(SEQ ID NO:4), KPRRPYTDNCTRLRKQMAVKKYLNSILN(SEQ ID NO:5), KPRRPYTDNYTSLRKQMAVKKYLNSILN(SEQ ID NO:6), KPRRPYTDNYTRLRKQIAVKKYLNSILN(SEQ ID NO:7), KPRRPYTDNYTRLRKQMAVNKYLNSILN(SEQ ID NO:8), KPRRPYTDNYTRLRKQMAVKKYMNSILN (SEQ ID NO:9), or KPRRPYTDNYTRLRKQMAVKKYLNLILN (SEQ ID NO:10).

[0011] In certain embodiments, the amino, carboxyl, hydroxyl, or thiol groups of the peptides disclosed herein are substituted. In certain embodiments, the peptides are attached to nanoparticles. In certain embodiments, the present disclosure contemplates the peptides disclosed herein having a label, e.g., fluorescent or radioactive.

[0012] In certain embodiments, the present disclosure relates to compositions, such as pharmaceutical compositions, and cell growth media comprising the peptides disclosed herein. In certain embodiments, the present disclosure relates to pharmaceutical compositions comprising the peptides disclosed herein and pharma- ceutically acceptable excipients. In certain embodiments, the pharmaceutical composition is in the form of a capsule, tablet, pill, powder, or granules. In certain embodiments, the pharmaceutical composition is in the form of a sterile pH-buffered aqueous salt solution. In certain embodiments, the pharmaceutical composition is in the form of a container configured to spray a liquid or a sealed container having a spray agent.

[0013] In certain embodiments, the present disclosure relates to a nucleic acid encoding a peptide disclosed herein in operable combination with a promoter. In certain embodiments, the present disclosure relates to a recombinant vector comprising a nucleic acid encoding a peptide disclosed herein in operable combination with a promoter. In certain embodiments, the present disclosure relates to an expression system or cell comprising a recombinant vector disclosed herein.

[0014] In certain embodiments, the present disclosure relates to a method of treating or enhancing an immune response against cancer or treating cancer, comprising administering to a subject in need thereof an effective amount of a peptide disclosed herein, hi certain embodiments, the peptide is administered in combination with another chemotherapeutic agent.

[0015] In certain embodiments, the present disclosure relates to a method of increasing T cell activity and ex vivo expansion, comprising mixing T cells with a peptide disclosed herein. In certain embodiments, the mixing of T cells is combined with an anti-CD3 antibody and / or an anti-CD28 antibody. In certain embodiments, the mixing of T cells is combined with a phosphatidylinositol 3-kinase delta (PI3Kδ) inhibitor.

[0016] In certain embodiments, the present disclosure relates to a method of treating or preventing host-versus-graft disease in a subject, comprising administering an effective amount of a peptide disclosed herein to a subject who is to receive, or has been administered, transplanted allogeneic tissue or cells.

[0017] In certain embodiments, the present disclosure relates to methods of treating cancer comprising exposing a subject to radiation and / or administering a chemotherapeutic agent to the subject, transplanting allogeneic hematopoietic stem cells to the subject, administering a peptide disclosed herein to the subject.

[0018] In certain embodiments, the present disclosure relates to a method of managing a viral infection comprising administering to a subject in need thereof an effective amount of a peptide disclosed herein.

[0019] In certain embodiments, the subject is a mammal, typically a human. [Brief description of the drawings]

[0020] [Figure 1]FIG. 1 shows antagonistic peptides that differ from VIPhyb of SEQ ID NO:1 by internal amino acids. Native VIP is SEQ ID NO:2. The sequences are named ANT-1-8 depending on the presence or absence of an internal amino acid (labeled "ANT"). In particular, ANT-1 has an amino acid substitution of T to A at amino acid position 7, SEQ ID NO:3. The ANT-2 sequence has an amino acid substitution of D to V at amino acid position number 8, SEQ ID NO:4. The ANT-3 peptide has an amino acid substitution of Y to C at amino acid position number 10, SEQ ID NO:5. The ANT-4 sequence has an amino acid substitution of R to S at amino acid position number 12, SEQ ID NO:6. The ANT-5 sequence has an amino acid substitution of M to I at amino acid position number 17, SEQ ID NO:7. The ANT-6 sequence has an amino acid substitution of K to N at amino acid position number 20, SEQ ID NO:8. The ANT-7 sequence has an L to M amino acid substitution at amino acid position 23, SEQ ID NO:9. The ANT-8 sequence has an S to L amino acid substitution at amino acid position 25, SEQ ID NO:10. [Diagram 2] Figure 2 shows data on T cell proliferation over 24 hours in the presence of original VIPhyb and Ant1 to Ant8. T cells from luciferase+ C57 / BL6 mice were collected and cultured in 96-well plates with 1 μg / ml anti-CD3 antibody and 30 U / ml IL-2 in the presence of 1 μM original VIPhyb, Ant-1, Ant-2, Ant-3, Ant-4, Ant-5, Ant-6, Ant-7 or Ant-8. [Diagram 3] Figure 3 shows data on T cell proliferation over 24 hours in the presence of original VIPhyb and Ant8. T cells from luciferase+ C57 / BL6 mice were harvested and cultured in 96-well plates with either 0.5 μg / ml or 1 μg / ml anti-CD3 antibody and 30 U / ml IL-2 in the presence of 0.5 μM, 1 μM or 3 μM original VIPhyb or Ant-8. [Figure 4]FIG. 4 shows data from mice subcutaneously injected with VIPhyb, Ant8, or PBS for 7 days following intravenous inoculation with leukemia cells. [Diagram 5] FIG. 5 shows survival data for leukemia-bearing mice (C1498) treated with ANT-8. [Figure 6A] Figure 6A shows data from immunocompetent mice injected with mouse luciferase transfected pancreatic cancer cell lines into the tail of the pancreas using KPC luc and treated with Ant-08 and / or PD1. A slower tumor growth rate was observed in mice treated with Ant-08 + anti-PD1. [Figure 6B] Figure 6B shows data on tumor burden in orthotopic KPC model. At the time of sacrifice (day 24), the tumor burden of the pancreas isolated from the above mice was weighed. Significantly smaller tumor burden was observed in mice treated with Ant-08+anti-PD1. [Figure 6C] Figure 6C shows data on CD4+ T cell infiltration in the orthotopic KPC model. Increased infiltration of CD4+ T cells was observed in tumors from mice treated with Ant-08 + anti-PD1. [Figure 6D] Figure 6D shows IVIS and MRI images of mice on day 26. Tumors from these mice were stained with H&E and CD4. [Figure 7] FIG. 7 shows data using ANT8 demonstrating increased yield of T cells from chronic lymphocytic leukemia (CLL) donors during ex vivo expansion with anti-CD3 / 28 beads and 30 U / mL IL-2. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0021] Before describing the present disclosure in more detail, it is to be understood that the present disclosure is not limited to particular embodiments described, which may, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting, as the scope of the present disclosure will be limited only by the appended claims.

[0022] Unless otherwise defined, 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. Although any methods and materials similar or equivalent to those described herein can also be used in the practice or testing of this disclosure, preferred methods and materials are described.

[0023] All publications and patents mentioned herein are incorporated by reference herein to the same extent as if each individual publication or patent was specifically and individually indicated to be incorporated by reference to describe the methods and / or materials in connection with which the publications are described. The mention of any publication is for its prior disclosure prior to the filing date and should not be construed as an admission that the present disclosure is not entitled to antedate such publication by virtue of prior disclosure. Further, the dates of publication provided may be different from the actual publication dates which may need to be independently confirmed.

[0024] As will be apparent to one of ordinary skill in the art upon reading this disclosure, each of the individual embodiments described and illustrated herein has separate elements and features which may be readily separated or combined with the features of any of the other several embodiments without departing from the scope or spirit of the disclosure. Any method described can be carried out in the order of events described or in any other order which is logically possible.

[0025] Embodiments of the present disclosure employ, unless otherwise indicated, techniques of immunology, pharmacology, organic chemistry, biochemistry, molecular biology, pharmacology, physics, and the like, which are within the skill of the art and which are fully explained in the literature.

[0026] It should be noted that as used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural references unless the context clearly dictates otherwise. In this specification and the following claims, reference is made to certain terms that are defined to have the following meanings, unless a contrary intention is apparent.

[0027] The terms "protein" and "peptide" refer to compounds that contain amino acids joined via peptide bonds and are used interchangeably. The amino acids may be naturally occurring or non-naturally occurring. A "chimeric protein" or "fusion protein" is a molecule in which different portions of a protein are derived from different sources and the entire molecule is not naturally occurring. A chimeric protein may contain amino acid sequences from the same species of different species, so long as they are not aligned together as they would be in nature. Examples of chimeric proteins include sequences disclosed herein that contain one, two or more amino acids attached to the C-terminus or N-terminus that are not identical to any naturally occurring protein, such as amino acids that contain amine side groups, e.g., lysine, amino acids that contain carboxylic acid side groups, such as aspartic acid or glutamic acid, polyhistidine tags, e.g., adding four or more histidine amino acids, as is typical.

[0028] The term "comprising" with reference to a peptide having an amino acid sequence refers to a peptide that may include additional N-terminal (amine end) or C-terminal (carboxylic acid end) amino acids, i.e., the term is intended to include the amino acid sequence in a larger peptide. The term "consisting of" with reference to a peptide having an amino acid sequence refers to a peptide having a precise number of amino acids or less, or only a range of amino acids explicitly specified in a claim. In certain embodiments, the present disclosure contemplates that "the N-terminus of the peptide can consist of an amino acid sequence," which refers to the N-terminus of a peptide having a precise number of amino acids or less, or only a range of amino acids explicitly specified in a claim, however, the C-terminus can be joined to additional amino acids, e.g., as part of a larger peptide. Similarly, the present disclosure contemplates that "the C-terminus of the peptide can consist of an amino acid sequence," which refers to the C-terminus of a peptide having a precise number of amino acids or less, or only a range of amino acids explicitly specified in a claim, however, the N-terminus can be joined to additional amino acids, e.g., as part of a larger peptide.

[0029] In certain embodiments, the present disclosure relates to recombinant peptides or fusions thereof comprising the sequences disclosed herein, where the amino or carbon terminus of the amino acid sequence is optionally attached to a heterologous amino acid sequence, a label, or a reporter molecule. "Label" refers to a detectable compound or composition that binds directly or indirectly to another molecule, such as an antibody or a protein, to facilitate detection of the molecule. Specific non-limiting examples of labels include fluorescent tags, enzyme conjugation, and radioactive isotopes. In one example, "labeled receptor" refers to the incorporation of a heterologous polypeptide into a receptor. Labels include the incorporation of a radiolabeled amino acid or the covalent attachment of a biotinyl moiety to a polypeptide that can be detected by labeled avidin (e.g., streptavidin containing a fluorescent marker or an enzymatic activity that can be detected by optical or colorimetric methods). Various methods of labeling polypeptides and glycoproteins are well known in the art and can be used. Examples of labels for polypeptides include, but are not limited to, the following: radioisotopes or radionuclides ( 35 S or 131 I, etc. fluorescent labels (fluorescein isothiocyanate (FITC), rhodamine, lanthanide fluorophores, etc.), enzyme labels (horseradish peroxidase, β-galactosidase, luciferase, alkaline phosphatase), chemiluminescent markers, biotinyl groups, defined peptide epitopes recognized by secondary reporters (leucine zipper pair sequences, binding sites for secondary antibodies, metal binding domains, epitope tags), or magnetic agents such as gadolinium chelators. In some embodiments, labels are attached by spacer arms of various lengths to reduce potential steric hindrance.

[0030] As used herein, the term "derivative" refers to a structurally similar peptide that retains sufficient functional attributes of the identified analog. A derivative may be structurally similar because it lacks one or more atoms, for example, an amino group, a hydroxyl, or a thiol group is replaced with hydrogen, a salt substituted with a different hydration / oxidation state, or because one or more atoms in the molecule are switched, such as, but not limited to, replacing an oxygen atom with a sulfur atom or replacing an amino group with a hydroxyl group. A derivative may be a prodrug and may include lipids, polyethylene glycols, sugars, polysaccharides. A derivative may be two or more peptides linked together by a linking group. It is contemplated that the linking group may be biodegradable. The derivatives can be prepared by any of the various synthetic methods or suitable adaptations of the synthesis provided in March's Advanced Organic Chemistry: Reactions, Mechanisms, and Structure, Wiley, 6th Edition (2007) Michael B. Smith or Domino Reactions in Organic Synthesis, Wiley (2006) Lutz F. Tietze, incorporated by reference, or shown in organic chemistry textbooks.

[0031] In certain embodiments, the peptides disclosed herein may be modified with, for example, polyethylene glycol attachment, attachment of another peptide resulting in a chimeric peptide, fluorescent dyes containing aromatic groups, fluorescent peptides, 18The peptides have at least one non-naturally occurring molecular modification, such as a chelator capable of binding a radionuclide such as F, an N-terminal acetyl, propionyl group, myristoyl and palmitoyl group, or an N-terminal methylation, or a C-terminal alkyl ester attachment. In certain embodiments, the present disclosure contemplates peptides disclosed herein that are labeled using commercially available biotinylation reagents. Biotinylated peptides can be used for streptavidin affinity binding, purification, and detection. In certain embodiments, the present disclosure contemplates peptides disclosed herein that include azido derivatives of naturally occurring monosaccharides, such as N-azidoacetylglucosamine and N-azidoacetylgalactosamine.

[0032] In certain embodiments, the present disclosure contemplates derivatives of the peptides disclosed herein, where one or more amino acids are replaced with a chemical group to improve pharmacokinetic properties such as solubility and serum half-life, and optionally connected by a linker. In certain embodiments, such derivatives may be prodrugs, where the substituent or linker is biodegradable, or where the substituent or linker is not biodegradable. In certain embodiments, contemplated substituents include sugars, polysaccharides, acetyl, fatty acids, lipids, and / or polyethylene glycol. The substituent may be covalently attached by forming an amide bond at the C-terminus or N-terminus of the peptide, optionally connected by a linker. In certain embodiments, it is contemplated that the substituent may be covalently attached by an amino acid in the peptide, for example, by an amine side group, such as lysine or an amino acid containing a carboxylic acid side group, such as aspartic acid or glutamic acid, in a peptide comprising a sequence disclosed herein. In certain embodiments, it is contemplated that the substituent may be covalently attached by a cysteine ​​in a sequence disclosed herein, which may be connected by a linker. In certain embodiments, the substituents are connected by a linker that forms a disulfide with a cysteine ​​amino acid side group.

[0033] The term "substituted" refers to a molecule in which at least one hydrogen atom is replaced with a substituent. When substituted, one or more of the group is a "substituent." The molecule may be multiply substituted. In the case of an oxo substituent ("=O"), two hydrogen atoms are replaced. Exemplary substituents in this context include halogen, hydroxy, alkyl, alkoxy, nitro, cyano, oxo, carbocyclyl, carbocycloalkyl, heterocarbocyclyl, heterocarbocycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, -NRaRb, -NRaC(=O)Rb, -NRaC(=O)NRaNRb, -NRaC(=O)ORb, -NRaSO 2 Rb, -C(=O)Ra, -C(=O)ORa, -C(=O)NRaRb, -OC(=O)NRaRb, -ORa, -SRa, -SORa, -S(=O) 2 Ra, -OS(=O) 2 Ra and -S(=O) 2 ORa can be mentioned. Ra and Rb in this context can be the same or different and can be independently hydrogen, halogen, hydroxyl, alkyl, alkoxy, alkyl, amino, alkylamino, dialkylamino, carbocyclyl, carbocycloalkyl, heterocarbocyclyl, heterocarbocycloalkyl, aryl, arylalkyl, heteroaryl, and heteroarylalkyl. The substituent can be further substituted.

[0034] As used herein, a "lipid" group refers to a naturally occurring or non-naturally occurring hydrophobic group that is highly insoluble in water. As used herein, a lipid group refers to a compound in which the lipid attachment points are replaced with hydrogen, resulting in a compound that is soluble by weight in water at a concentration of 0.63x10 -4A lipid is considered to be highly water insoluble when it has a solubility in water of less than 100% w / w (25°C). See Solvent Recovery Handbook, 2nd Edition, Smallwood, 2002 by Blackwell Science, p. 195. Examples of naturally occurring lipids include the saturated or unsaturated hydrocarbon chains found in fatty acids, glycerolipids, cholesterol, steroids, polyketides and derivatives. Non-naturally occurring lipids include naturally occurring lipids, acrylic polymers, aromatic, and alkylated compounds and their derivatives.

[0035] The term "prodrug" refers to an agent that is converted in vivo into a biologically active form. Prodrugs are often useful because in some situations they may be easier to administer than the parent compound. Prodrugs may also have improved solubility in pharmaceutical compositions over the parent drug. Prodrugs may be converted to the parent drug by various mechanisms, including enzymatic processes and metabolic hydrolysis. Exemplary prodrugs are pharma-ceutically acceptable esters. Prodrugs include compounds in which a hydroxy, amino, or mercapto (thiol) group is bonded to any group that cleaves to form a free hydroxy, free amino, or free mercapto group, respectively, when the prodrug of the active compound is administered to a subject. Examples of prodrugs include, but are not limited to, acetate, formate, and benzoic acid derivatives of alcohols or acetamides, as well as formamide and benzamide derivatives of amine functional groups of such active compounds.

[0036] For example, if a disclosed peptide or a pharma- ceutically acceptable form of a peptide contains a carboxylic acid functional group, the prodrug may be a prodrug that replaces the hydrogen atom of the acidic group with (C 1 -C 8 ) alkyl, (C 2 -C 12)alkanoyloxymethyl, 1-(alkanoyloxy)ethyl having 4 to 9 carbon atoms, 1-methyl-1-(alkanoyloxy)-ethyl having 5 to 10 carbon atoms, alkoxycarbonyloxymethyl having 3 to 6 carbon atoms, 1-(alkoxycarbonyloxy)ethyl having 4 to 7 carbon atoms, 1-methyl-1-(alkoxycarbonyloxy)ethyl having 5 to 8 carbon atoms, N-(alkoxycarbonyl)aminomethyl having 3 to 9 carbon atoms, 1-(N-(alkoxycarbonyl)amino)ethyl having 4 to 10 carbon atoms, 3-phthalidyl, 4-crotonolactonyl, gamma-butyrolactone-4-yl, di-N,N-(C 1 -C 2 ) Alkylamino(C 2 -C 3 ) alkyl (beta-dimethylaminoethyl, etc.), carbamoyl-(C 1 -C 2 ) alkyl, N,N-di(C 1 -C 2 ) alkylcarbamoyl-(C 1 -C 2 ) alkyl and piperidino-, pyrrolidino- or morpholino (C 2 -C 3 ) alkyl or the like.

[0037] When the disclosed peptide or pharma- ceutically acceptable form of the peptide contains an alcohol functional group, the prodrug may be a prodrug that converts the hydrogen atom of the alcohol group to (C 1 -C 6 ) alkanoyloxymethyl, 1-((C 1 -C 6 )alkanoyloxy)ethyl, 1-methyl-1((C 1 -C 6 )Alkanoyloxy)ethyl(C 1 -C 6 ) alkoxycarbonyloxymethyl, -N-(C 1 -C 6 ) alkoxycarbonylaminomethyl, succinoyl, (C 1 -C 6) Alkanoyl, alpha-amino (C 1 -C 4 ) alkanoyl, arylacyl and alpha-aminoacyl, or alpha-aminoacyl-alpha-aminoacyl groups, each of which independently represents a naturally occurring L-amino acid P(O)(OH) 2 , -P(O)(O(C 1 -C 6 )Alkyl) 2 and glycosyl (the radical resulting from removal of the hydroxyl group of the hemiacetal form of a carbohydrate).

[0038] When a disclosed peptide or a pharma- ceutically acceptable form of a peptide incorporates an amine functional group, a prodrug can be formed by replacing the hydrogen atom of the amine group with a group such as R-carbonyl, RO-carbonyl, NRR′-carbonyl, or the like, where R and R′ are each independently (C 1 -C 10 ) alkyl, (C 3 -C 7 ) Cycloalkyl, benzyl, natural alpha-aminoacyl, -C(OH)C(O)OY 1 and Y 1 is H, (C 1 -C 6 ) alkyl or benzyl, -C(OY 2 )Y 3 and Y 2 (C 1 -C 4 ) alkyl, and Y 3 (C 1 -C 6 ) alkyl, carboxy (C 1 -C 6 ) Alkyl, Amino (C 1 -C 4 ) alkyl or mono-N or di-N,N-(C 1 -C 6 ) alkylaminoalkyl, -C(Y 4 )Y 5 and Y 4 is H or methyl, Y 5is mono-N- or di-N,N-(C 1 -C 6 ) alkylamino, morpholino, piperidin-1-yl or pyrrolidin-1-yl.

[0039] As used herein, "pharmacologically acceptable esters" include, but are not limited to, alkyl, alkenyl, alkynyl, aryl, arylalkyl, and cycloalkyl esters of acidic groups, including, but not limited to, carboxylic acids, phosphoric acids, phosphinic acids, sulfonic acids, sulfinic acids, and boronic acids.

[0040] As used herein, "pharmacologically acceptable enol ethers" include, but are not limited to, derivatives of the formula -C=C(OR), where R can be selected from alkyl, alkenyl, alkynyl, aryl, aralkyl, and cycloalkyl. Pharmaceutically acceptable enol esters include, but are not limited to, derivatives of the formula -C=C(OC(O)R), where R can be selected from hydrogen, alkyl, alkenyl, alkynyl, aryl, aralkyl, and cycloalkyl.

[0041] "Linking group" refers to any of a variety of molecular arrangements that can be used to link molecular moieties together. An exemplary formula is -R m R may be -CR m R m -, -CHR m -, -CH-, -C-, -CH 2 -, -C(OH)R m , -C(OH)(OH)-, -C(OH)H, -C(Hal)R m -, -C(Hal)(Hal)-, -C(Hal)H-, -C(N 3 )R m -, -C(CN)R m -, -C(CN)(CN)-, -C(CN)H-, -C(N 3 )(N 3 )-, -C(N 3 )H-, -O-, -S-, -N-, -NH-, -NR m-, -(C=O)-, -(C=NH)-, -(C=S)-, -(C=CH 2 )-, at each occurrence, is individually and independently selected and may individually and independently include a single bond, double bond, or triple bond between the R groups. m If branched at -CH 3 , -H, -CH=CH 2 , -CCH, -OH, -SH, -NH 2 , -N 3 Rs may terminate in a group such as -CN or -Hal, or two branched Rs may form a cyclic structure. In certain instances, it is contemplated that the total Rs or "m" may be less than 100, or 50, or 25, or 10. Examples of linking groups include bridging alkyl groups and alkoxyalkyl groups. The linking groups may be substituted with one or more substituents.

[0042] The term "nucleic acid" refers to a polymer of nucleotides, or polynucleotide. This term is used to indicate a molecule or a collection of molecules. A nucleic acid may be single-stranded or double-stranded and may contain a coding region and various regulatory element regions, as described below.

[0043] The term "nucleic acid sequence encoding" a particular peptide refers to a nucleic acid sequence that comprises a coding region for the peptide, or in other words, a nucleic acid sequence that encodes a peptide product. The coding region may be present in either cDNA, genomic DNA or RNA form. If present in DNA form, the oligonucleotide, polynucleotide or nucleic acid may be single-stranded (i.e., the sense strand) or double-stranded. Appropriate control elements such as enhancers / promoters, splice junctions, polyadenylation signals, etc. may be placed in close proximity to the coding region as necessary to allow proper initiation of transcription and / or correct processing of the primary RNA transcript. Alternatively, the coding region used in an expression vector may contain endogenous enhancers / promoters, splice junctions, intervening sequences, polyadenylation signals, etc. or a combination of both endogenous and exogenous control elements.

[0044] The term "vector" or "expression vector" refers to a recombinant nucleic acid that contains a desired coding sequence and appropriate nucleic acid sequences necessary for expression of an operably linked coding sequence in a particular host organism or expression system, e.g., cellular or cell-free. Nucleic acid sequences necessary for expression in prokaryotes usually include a promoter, an operator (optional), and a ribosome binding site, often along with other sequences. Eukaryotic cells are known to use promoters, enhancers, and termination and polyadenylation signals.

[0045] Protein "expression systems" refer to in vivo and in vitro (cell-free) systems. Systems for recombinant protein expression usually use cells that are transfected with a DNA expression vector containing a template. The cells are cultured under conditions that translate the desired protein. The expressed protein is extracted for subsequent purification. In vivo protein expression systems using prokaryotic and eukaryotic cells are well known. Some proteins are also recovered using denaturants and protein refolding procedures. In vitro (cell-free) protein expression systems usually use translation-compatible extracts of whole cells or compositions that contain sufficient components for transcription, translation, and any post-translational modifications, such as RNA polymerase, regulatory protein factors, transcription factors, ribosomes, tRNA cofactors, amino acids, and nucleotides. In the presence of an expression vector, these extracts and components can synthesize the protein of interest. Cell-free systems usually do not contain proteases and can label proteins with modified amino acids. Some cell-free systems have incorporated components coded for translation into expression vectors. See, e.g., Shimizu et al., Cell-free translation reconstituted with purified components, 2001, Nat. BioTechnol., 19, 751-755, and Asahara & Chong, Nucleic Acids Research, 2010, 38(13):e141, both of which are incorporated by reference in their entireties.

[0046] "Cancer" refers to any of a variety of cellular diseases with malignant tumors characterized by cell proliferation. It is not intended that the cells of the disease must actually invade surrounding tissues and metastasize to new body sites. Cancer can involve any tissue of the body and can have many different forms in each body area. In the context of certain embodiments, whether "cancer is reduced" can be identified by various diagnostic modalities known to those skilled in the art, including but not limited to observing a reduction in tumor mass or number, or observing an increase in apoptosis of cancer cells, for example, observing a greater than 5% increase in apoptosis of cancer cells for a sample compound compared to a control without the compound. It can also be identified by changes in relevant biomarkers or gene expression profiles, such as PSA for prostate cancer, HER2 for breast cancer, or others.

[0047] The cancer to be treated in the context of this disclosure may be any type of cancer or tumor. These tumors or cancers include, but are not limited to, tumors of hematopoietic and lymphoid tissues, or hematopoietic and lymphoid malignancies, tumors affecting the blood, bone marrow, lymph, and lymphatic systems. Hematopoietic malignancies may originate from either of two major blood cell lineages: myeloid cell lineages and lymphoid cell lineages. Myeloid cell lineages usually produce granulocytes, red blood cells, platelets, macrophages, and mast cells, while lymphoid cell lineages produce B, T, NK, and plasma cells. Lymphomas, lymphocytic leukemias, and myelomas originate from lymphoid lineages, while acute and chronic myeloid leukemias, myelodysplastic syndromes, and myeloproliferative disorders originate from the bone marrow.

[0048] Tumors located in the colon, abdomen, bone, breast, digestive system, liver, pancreas, peritoneum, endocrine glands (adrenal, parathyroid, pituitary, testes, ovaries, thymus, thyroid), eye, head and neck, nervous system (central and peripheral), lymphatic system, pelvis, skin, soft tissue, spleen, chest, and genitourinary tract are also contemplated, more particularly childhood acute lymphoblastic leukemia, acute lymphoblastic leukemia, acute lymphocytic leukemia, acute myeloid leukemia, adrenal cortical carcinoma, adult (primary) hepatocellular carcinoma, adult (primary) Liver cancer, adult acute lymphocytic leukemia, adult acute myeloid leukemia, adult Hodgkin's disease, adult Hodgkin's lymphoma, adult lymphocytic leukemia, adult non-Hodgkin's lymphoma, adult primary liver cancer, adult soft tissue sarcoma, AIDS-related lymphoma, AIDS-related malignant tumor, anal cancer, astrocytoma, cancer of the biliary tract, cancer of the bladder, bone cancer, brain stem glioma, brain tumor, breast cancer, cancer of the renal pelvis and ureter, primary central nervous system lymphoma, central nervous system lymphoma, cerebellar astrocytoma, brain astrocytoma, cancer of the cervix cervix), childhood (primary) hepatocellular carcinoma, childhood (primary) liver cancer, childhood acute lymphoblastic leukemia, childhood acute myeloid leukemia, childhood brain stem glioma, childhood cerebellar astrocytoma, childhood brain astrocytoma, childhood extracranial germ cell tumor, childhood Hodgkin's lymphoma, childhood Hodgkin's lymphoma, childhood visual pathway and hypothalamic glioma, childhood lymphocytic leukemia, childhood medulloblastoma, childhood non-Hodgkin's lymphoma, childhood supratentorial primitive neuroectodermal and pineal tumor, childhood primary liver cancer, childhood rhabdomyosarcoma, childhood soft tissue sarcoma, childhood visual pathway and hypothalamic glioma, chronic Lymphocytic leukemia, chronic myeloid leukemia, colon cancer, cutaneous T-cell lymphoma, endocrine islet cell carcinoma, endometrial cancer, ependymoma, epithelial carcinoma, esophageal cancer, Ewing's sarcoma and related tumors, cancer of the exocrine pancreas, extracranial germ cell tumors, extragonadal germ cell tumors, extrahepatic bile duct cancer, eye cancer, female breast cancer, Gaucher's disease, gallbladder cancer, gastric cancer, gastrointestinal carcinoids, gastrointestinal tumors, germ cell tumors, gestational trophoblastic tumors, head and neck cancer, hepatocellular carcinoma, Hodgkin's disease, Hodgkin's lymphoma, hypergammaglobulinemia, hypopharyngeal cancer, intestinal cancer, intraocular melanoma, islet cell carcinoma, islet cell pancreatic cancer, Kaposi's sarcoma, kidney cancer, laryngeal cancer, cancer of the lip and lipmouth, liver cancer, lung cancer, lymphoproliferative disorders, macroglobulinemia, male breast cancer, malignant mesothelioma, malignant thymoma, medulloblastoma, melanoma, mesothelioma, occult primary metastatic squamous neck cancer, primary metastatic squamous neck cancer, metastatic squamous neck cancer, multiple myeloma, multiple myeloma / plasmatic cell neoplasia, myelodysplastic syndromes, myeloid leukemia, myeloproliferative disorders, paranasal sinus and nasal cancer, nasopharyngeal carcinoma, neuroblastoma, non-Hodgkin's lymphoma during pregnancy, non-melanoma skin cancer, non-small cell lung cancer, metastatic squamous neck cancer with occult primary primary), buccopharyngeal cancer, malignant fibrous histiocytoma, malignant fibrous osteosarcoma / histiocytoma of bone, epithelial ovarian cancer, ovarian germ cell tumor, ovarian low malignant potential tumor, pancreatic cancer, paraproteinemia, purpura, parathyroid cancer, penile cancer, pheochromocytoma, pituitary tumor (hypophysis tumor), neoplasms of plasma cells / multiple myeloma, primary central nervous system lymphoma, primary liver cancer, prostate cancer, rectal cancer, renal cell carcinoma, cancer of the renal pelvis and ureter, retinoblastoma, rhabdomyosarcoma, cancer of the salivary glands, sarcoidosis, sarcoma, skin cancer, small cell lung cancer, small intestine cancer, soft tissue sarcoma, squamous cell cervical cancer, gastric cancer, pineal and supratentorial primary neuroectodermal tumor, T-cell lymphoma, testicular cancer, thymoma, thyroid cancer, transitional cell carcinoma of the renal pelvis and ureter, transitional renal pelvis and ureter cancer, trophoblastic tumor, cancer of the renal pelvis and ureter, cancer of the urethra, uterine cancer, uterine sarcoma, vaginal cancer, visual pathway and hypothalamic glioma, cancer of the vulva, Waldenstrom's hypergammaglobulinemia, Wilms' tumor and any other hyperproliferative disease and tumor are also contemplated.

[0049] "Chemotherapeutic agent", "chemotherapeutic drug", "anticancer agent" and the like refer to molecules recognized to aid in the treatment of cancer. Contemplated examples include the following molecules or derivatives: alemtuzumab, trastuzumab, ibritumomab tiuxetan, brentuximab vedotin, temozolomide, ado-trastuzumab emtansine, denileukin diftitox, blinatumomab, interferon alpha, aldesleukin, carmustine, bevacizumab, procarbazine, lomustine, vincristine, gefitinib, erlotinib, cisplatin, carboplatin, oxaliplatin, 5-fluorouracil, gemcitabine, tegafur, raltitrexed, methotrexate, cytosine arabinoside, hydroxyurea, adriamycin, bleomycin, Doxorubicin, daunomycin, epirubicin, idarubicin, mitomycin-C, dactinomycin, mithramycin, vinblastine, vindesine, vinorelbine, paclitaxel, taxol, docetaxel, etoposide, teniposide, amsacrine, topotecan, camptothecin, bortezomib, anagrelide, tamoxifen, toremifene, raloxifene, droloxifene, fulvestrant, bicalutamide, flutamide, nilutamide, Cyproterone, goserelin, leuprorelin, buserelin, megestrol, anastrozole, letrozole, vorozole, exemestane, finasteride, marimastat, trastuzumab, cetuximab, dasatinib, imatinib, combretastatin, thalidomide, azacitidine, azathioprine, capecitabine, chlorambucil, cyclophosphamide, cytarabine, daunorubicin, doxifluridine, epothilone, irinotecan, mechlorethamine, tamoxifen, mercaptopurine, mitoxantrone, pemetrexed, thioguanine, valrubicin, rituximab, and / or lenalidomide or for example, cyclophosphamide, methotrexate, 5-fluorouracil (CMF); doxorubicin, cyclophosphamide (AC); mustine, vincristine, procarbazine, prednisolone (MOPP); adriamycin, bleomycin, vinblastine, dacarbazine (ABVD);Cyclophosphamide, doxorubicin, vincristine, prednisolone (CHOP); rituximab, cyclophosphamide, doxorubicin, vincristine, prednisolone (RCHOP); bleomycin, etoposide, cisplatin (BEP); epirubicin, cisplatin, 5-fluorouracil (ECF); epirubicin, cisplatin, capecitabine (ECX); methotrexate, vincristine, doxorubicin, cisplatin (MVAC) and combinations thereof.

[0050] In certain embodiments, the cancer treatment can be combined with another anti-cancer drug. In certain embodiments, the anti-cancer drug can be abemaciclib, abiraterone acetate, methotrexate, paclitaxel, adriamycin, acalabrutinib, brentuximab vedotin, ado-trastuzumab emtansine, aflibercept, afatinib, netupitant, palonosetron, imiquimod, aldesleukin, alectinib, alemtuzumab, pemetrexed disodium, copanlisib, melphalan, brigatinib, chlorambucil, amifostine, aminolevulinic acid, anastrozole, apalutamide, Aprepitant, pamidronate disodium, exemestane, nelarabine, arsenic trioxide, ofatumumab, atezolizumab, bevacizumab, avelumab, axicaptagene ciloreucel, axitinib, azacitidine, carmustine, belinostat, bendamustine, inotuzumab ozogamicin, bevacizumab, bexarotene, bicalutamide, bleomycin, blinatumomab, bortezomib, bosutinib, brentuximab vedotin, brigatinib, busulfan, irinotecan, capecitabine, fluorouracil, carbo Platinum, carfilzomib, ceritinib, daunorubicin, cetuximab, cisplatin, cladribine, cyclophosphamide, clofarabine, cobimetinib, cabozantinib-S-malate, dactinomycin, crizotinib, ifosfamide, ramucirumab, cytarabine, dabrafenib, dacarbazine, decitabine, daratumumab, dasatinib, defibrotide, degarelix, denileukin diftitox, denosumab, dexamethasone, dexrazoxane, dinutuximab, docetaxel, doxorubicin, durval Mab, rasburicase, epirubicin, elotuzumab, oxaliplatin, eltrombopag olamine, enasidenib, enzalutamide, eribulin, vismodegib, erlotinib, etoposide, everolimus, raloxifene, toremifene, panobinostat, fulvestrant, letrozole, filgrastim, fludarabine, flutamide, pralatrexate, obinutuzumab, gefitinib, gemcitabine, gemtuzumab ozogamicin, glucarpidase, goserelin, propranolol, trastuzumab, topotecan,Palbociclib, Ibritumomab tiuxetan, Ibrutinib, Ponatinib, Idarubicin, Idelalisib, Imatinib, Talimogene laherparepvec, Ipilimumab, Romidepsin, Ixabepilone, Ixazomib, Ruxolitinib, Cabazitaxel, Palifermin, Pembrolizumab, Ribociclib, Tisagenlecleucel, Lanreotide, Lapatinib, Olaratumab, Lenalidomide, Lenvatinib, Leucovorin, Leuprolide, Lomustine, Trifluridine, Olaparib, Vincristine, Procarbazine, Mechlorethamine, Megestrol, Trametinib, Temozolomide, Methylnaltrexone bromide, Midostaurin, Mitomycin C, Mitoxantrone, Plerixafor, vinorelbine, necitumumab, neratinib, sorafenib, nilutamide, nilotinib, niraparib, nivolumab, tamoxifen, romiplostim, sonidegib, omacetaxine, pegaspargase, ondansetron, osimertinib, panitumumab, pazopanib, interferon alpha-2b, pertuzumab, pomalidomide, mercaptopurine, regorafenib, rituximab, rolapitant, rucaparib, siltuximab, sunitinib, thioguanine, temsirolimus, thalidomide, thiotepa, trabectedin, valrubicin, vandetanib, vinblastine, vemurafenib, vorinostat, zoledronic acid, or a combination thereof.

[0051] In certain embodiments, the method of administration is to a subject having a lymphodepleted environment, hi certain embodiments, the lymphodepleting agents are cyclophosphamide and fludarabine.

[0052] As used herein, the term "idelalisib" refers to the compound (S)-2-(1-(9H-purin-6-ylamino)propyl)-5-fluoro-3-phenylquinazolin-4(3H)-one or a substitute salt thereof.

[0053] As used herein, "T cells that are CD28 and / or CD27 negative" refers to T cells that have low or poor expression of the relative concentrations of these markers when compared to normal T cells that express the CD3 surface antigen marker in a healthy subject.

[0054] The term "fluorescence activated cell sorting" or "FACS" refers to a method of sorting a mixture of cells into two or more regions, usually one cell at a time, based on the fluorescent characteristics of each cell, the charge applied to each, and separation by movement through an electrostatic field. A vibration mechanism usually breaks the stream of cells into individual droplets. Just prior to droplet formation, the cells in the fluid pass through a region for measuring the fluorescence of the cells. A charging mechanism is configured at the point where the stream breaks into droplets. As the droplets break from the stream, a respective charge is imposed on the droplets based on the fluorescence intensity measurement. The charged droplets are then moved through an electrostatic deflector that diverts the droplets into regions based on their relative charge. In some systems, a charge is applied directly to the stream, and the break state of the droplets retains a charge of the same sign as the stream. The stream then returns to neutral after the droplets break off. In other systems, a charge is provided to a conduit that induces an opposite charge on the droplets. The cells are usually made fluorescent by mixing the cells with an antibody that specifically binds to a marker that becomes fluorescent by binding to a fluorescent molecule. However, other methods of making cells fluorescent, such as by using molecular beacons, are contemplated.

[0055] "Basal medium" refers to a medium that includes calcium, magnesium, potassium, sodium, phosphate, and bicarbonate, vitamins, and salts of essential amino acids. The 12 essential amino acids are L-arginine, L-cysteine, L-glutamine, L-histidine, L-isoleucine, L-leucine, L-methionine, L-phenylalanine, L-threonine, L-tryptophan, L-tyrosine, and L-valine. MEM is often supplemented with components such as bicarbonate or glutamine. In certain embodiments, the present disclosure also contemplates a basal medium supplemented with non-essential amino acids: L-ala, L-asn, L-asp, L-glu, L-gly, L-pro, and L-ser. In certain embodiments, the present disclosure contemplates a basal medium supplemented with nucleosides (ribonucleosides and / or deoxyribonucleosides).

[0056] The term "recombinant" when referring to a nucleic acid molecule refers to a nucleic acid molecule that is comprised of segments of nucleic acid joined together by molecular biological techniques. The term "recombinant" when referring to a protein or polypeptide refers to a protein molecule that is expressed using a recombinant nucleic acid molecule. The term recombinant nucleic acid is distinguished from natural recombination, which results from crossing over between homologous chromosomes. Recombinant nucleic acid as used herein is the non-natural combination of nucleic acids, usually from heterologous sources from different organisms.

[0057] The term "vector" or "expression vector" refers to a recombinant nucleic acid that contains a desired coding sequence and appropriate nucleic acid sequences necessary for expression of an operably linked coding sequence in a particular host organism or expression system, e.g., cellular or cell-free. Nucleic acid sequences necessary for expression in prokaryotes usually include a promoter, an operator (optional), and a ribosome binding site, often along with other sequences. Eukaryotic cells are known to use promoters, enhancers, and termination and polyadenylation signals.

[0058] The terms "vasoactive intestinal peptide" and "VIP" refer to (SEQ ID NO:2) HSDAVFTDNYTRLRKQMAVKKYLNSILN unless the context suggests otherwise. VIP is a multifunctional endogenous polypeptide that regulates both innate and adaptive immunity at multiple levels of immune cell differentiation and activity. VIP is normally secreted by a variety of cells, such as neuronal B-chain cells, T cells, and accessory cells (in both the central and limbic nervous systems). VIP and the closely related neuropeptide pituitary adenylate cyclase-activating polypeptide (PACAP) bind to three well-known receptors VPAC1, VPAC2, and PAC1. T cells and dendritic cells (DCs) are thought to express VPAC1 and VPAC2, but not PAC1. PAC1 is expressed primarily in neurons and endocrine cells in the brain and pituitary and adrenal glands, and in its most abundant form selectively binds PACAP.

[0059] "Subject" refers to any animal, preferably a human patient, livestock or household pet.

[0060] As used herein, the terms "prevent" and "preventing" include prevention of recurrence, spread, or onset. The disclosure is not intended to be limited to complete prevention. In some embodiments, onset is delayed or the severity of the disease is reduced.

[0061] As used herein, the terms "treat" and "treating" are not limited to cases where a subject (e.g., a patient) is cured or a disease is eradicated. Rather, embodiments of the present disclosure also contemplate treatment that merely reduces symptoms and / or slows the progression of a disease.

[0062] As used herein, the term "in combination with" is used to describe administration of an additional therapeutic means where the agent can be administered before, together with, or after the additional treatment, or in combination.

[0063] composition In certain embodiments, the present disclosure contemplates a pharmaceutical composition comprising a peptide disclosed herein, or a nanoparticle thereof, or optionally another pharmaceutical agent, or a pharma- ceutically acceptable salt thereof, and a pharma- ceutically acceptable excipient.

[0064] In certain embodiments, the present disclosure relates to compositions, such as pharmaceutical compositions, and cell growth media comprising the peptides disclosed herein. In certain embodiments, the present disclosure relates to pharmaceutical compositions comprising the peptides disclosed herein and pharma- ceutically acceptable excipients. In certain embodiments, the pharmaceutical composition is in the form of a capsule, tablet, pill, powder, or granules. In certain embodiments, the pharmaceutical composition is in the form of a sterile pH-buffered aqueous salt solution. In certain embodiments, the pharmaceutical composition is in the form of a container configured to spray a liquid or a sealed container having a spray agent.

[0065] Effective amounts can be extrapolated from dose-response curves derived from in vitro or animal model test systems. For the peptides or nanoparticles thereof, or other agents disclosed herein, dosages of 0.0001 mg / kg to 100 mg / kg of patient body weight are typically administered. Preferably, a dose of 0.0001 mg / kg to 20 mg / kg, 0.0001 mg / kg to 10 mg / kg, 0.0001 mg / kg to 5 mg / kg, 0.0001 to 2 mg / kg, 0.0001 to 1 mg / kg, 0.0001 mg / kg to 0.75 mg / kg, 0.0001 mg / kg to 0.5 mg / kg, 0.0001 mg / kg to 0.25 mg / kg, 0.0001 to 0.15 mg / kg, 0.0001 to 0.10 mg / kg, 0.001 to 0.5 mg / kg, 0.01 to 0.25 mg / kg or 0.01 to 0.10 mg / kg of the patient's body weight is administered. Furthermore, the dose and frequency of administration of the peptides or nanoparticles or drugs disclosed herein can be reduced by increasing uptake and tissue penetration through modifications such as lipidation and encapsulation in natural or artificial pulmonary surfactants.

[0066] Compositions include pharmaceutical compositions (e.g., impure or non-sterile compositions) and drug substance compositions useful for manufacturing pharmaceutical compositions (i.e., compositions suitable for administration to a subject or patient) that can be used to prepare unit dosage forms. Such compositions include a prophylactically or therapeutically effective amount of a prophylactic and / or therapeutic agent disclosed herein or a combination of such agents and a pharmaceutically acceptable carrier. In certain embodiments, the pharmaceutical composition includes a pharmaceutically acceptable excipient that is a solubilizing agent, such as a lipid, cholesterol, a fatty acid, a fatty acid alkyl ester, linoleic acid, oleic acid, arachidonic acid, a sugar, a polysaccharide, a cyclodextrin, 2-hydroxypropyl (cyclodextrin), or a combination thereof.

[0067] In certain embodiments, the pharma- ceutically acceptable excipient is selected from lactose, sucrose, mannitol, triethyl citrate, glucose, cellulose, methylcellulose, ethylcellulose, hydroxylpropylcellulose, hydroxypropylmethylcellulose, carboxymethylcellulose, croscarmellose sodium, polyvinyl N-pyrrolidone, crospovidone, ethylcellulose, povidone, methyl and ethyl acrylate copolymers, polyethylene glycol, fatty acid esters of sorbitol, lauryl sulfate, gelatin, glycerin, glyceryl monooleate, silicon dioxide, titanium dioxide, talc, corn starch, stearic acid, sorbic acid, magnesium stearate, calcium stearate, castor oil, mineral oil, calcium phosphate, starch, carboxymethyl ether of starch, iron oxide, triacetin, gum acacia, esters, or salts thereof.

[0068] In certain embodiments, the pharmaceutical composition is in a solid form surrounded by an enteric coating, i.e., a polymeric barrier applied to oral medication to prevent dissolution or disintegration in the intestinal environment. Compounds commonly found in enteric coatings include methyl acrylate-methacrylic acid copolymer, cellulose acetate phthalate (CAP), cellulose acetate succinate, hydroxypropyl methylcellulose phthalate, hydroxypropyl methylcellulose acetate succinate (hypromellose acetate succinate), polyvinyl acetate phthalate (PVAP), methyl methacrylic acid-methacrylic acid copolymer, and combinations thereof.

[0069] In certain embodiments, the term "pharmaceutical acceptable" means approved by a federal or state government regulatory agency or listed in the United States Pharmacopoeia or other generally recognized pharmacopoeias for use in animals and more particularly in humans. The term "carrier" refers to a diluent, adjuvant (e.g., Freund's adjuvant (complete and incomplete)), excipient, or vehicle with which the therapeutic is administered. Such pharmaceutical carriers can be sterile liquids of both natural and artificial pulmonary surfactants, water, and oils and sugars, including those of petroleum, animal, vegetable, or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil, and the like. Water is a preferred carrier when the pharmaceutical composition is administered intravenously. Saline solutions and aqueous dextrose and glycerol solutions can also be used as liquid carriers, particularly for injectable solutions. Suitable pharmaceutical excipients include starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk, glycerol, propylene, glycol, water, ethanol, etc. If desired, the compositions can also contain minor amounts of wetting or emulsifying agents, or pH buffering agents. These compositions can take the form of solutions, suspensions, emulsions, tablets, pills, capsules, powders, sustained-release formulations, and the like.

[0070] Generally, the components of the composition are supplied separately or mixed together in unit dosage form, for example as a lyophilized powder or water-free concentrate in a hermetically sealed container, such as an ampule indicating the quantity of active agent. When the composition is administered by infusion, it can be dispensed with an infusion bottle containing sterile pharmaceutical grade water or saline. When the composition is administered by injection, an ampoule of sterile water for injection or saline can be provided so that the components can be mixed prior to administration.

[0071] The compositions can be formulated as neutral or salt forms. Pharmaceutically acceptable salts include, but are not limited to, salts formed with anions such as those derived from hydrochloric acid, phosphoric acid, acetic acid, oxalic acid, tartaric acid, and the like, and salts formed with cations such as sodium, potassium, ammonium, calcium, ferric hydroxide, isopropylamine, triethylamine, 2-ethylaminoethanol, histidine, procaine, and the like.

[0072] An embodiment provides a pharmaceutical pack or kit comprising one or more containers filled with a peptide or nanoparticles thereof as disclosed herein or an agent as disclosed herein. Additionally, one or more other prophylactic or therapeutic agents useful for treating a disease may also be included in the pharmaceutical pack or kit. An embodiment provides a pharmaceutical pack or kit comprising one or more containers filled with one or more components of a pharmaceutical composition. Associated with such container(s) may be a notice in the form prescribed by a governmental agency regulating the manufacture, use or sale of pharmaceuticals or biological substances, which notice reflects approval by the period of manufacture, use or sale for administration to humans.

[0073] In certain embodiments, the present disclosure contemplates pharmaceutical compositions comprising the peptides or nanoparticles thereof disclosed herein, and agents disclosed herein and pharma- ceutical acceptable excipients. In certain embodiments, the present disclosure contemplates the manufacture of medicaments comprising the peptides or nanoparticles thereof disclosed herein, or agents disclosed herein, and uses for the methods disclosed herein.

[0074] In certain embodiments, the present disclosure relates to a pharmaceutical composition comprising a peptide or nanoparticle thereof disclosed herein, and an agent disclosed herein and a pharma- ceutical acceptable excipient. In certain embodiments, the composition is a pill or capsule, or the composition is an aqueous buffer, for example, at a pH of 6-8. In certain embodiments, the pharma- ceutical acceptable excipient is selected from a filler, a glidant, a binder, a disintegrant, a lubricant, and a sugar.

[0075] Compositions suitable for oral injection may include pharma- ceutically acceptable sterile aqueous or non-aqueous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Examples of suitable aqueous and non-aqueous carriers, diluents, solvents or vehicles include ethanol, polyols (propylene glycol, polyethylene glycol, glycerol, etc.), suitable mixtures thereof, vegetables (olive oil, sesame oil and viscoleo), preparations incorporated into pulmonary surfactants (both natural and synthetic), and injectable organic esters such as ethyl oleate.

[0076] Prevention of the action of microorganisms can be controlled by the addition of any of a variety of antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, etc. It may also be desirable to include isotonic agents, for example, sugars, sodium chloride, etc. Prolonged absorption of the injectable pharmaceutical form can be brought about by the use of agents delaying absorption, for example, aluminum monostearate and gelatin.

[0077] Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In such solid dosage forms, the peptides disclosed herein or nanoparticles thereof, or drugs, are mixed with at least one inert conventional excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with (a) fillers or extenders, such as starch, lactose, sucrose, glucose, mannitol and silicic acid, (b) binders, such as carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and acacia, (c) humectants, such as glycerol, (d) disintegrants, such as agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates, sodium carbonate, (e) solution inhibitors, such as glycerol, (f) glycerol, (g) glycerol, (h) glycerol, (i) glycerol, (j) glycerol, (k) glycerol, (l ... (f) absorption promoters, such as quaternary ammonium compounds, (g) wetting agents, such as cetyl alcohol, glycerol monostearate, (h) adsorbents, such as kaolin and bentonite, and (i) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, or mixtures thereof. In the case of capsules, tablets, and pills, the dosage form may also contain buffering agents.

[0078] Liquid dosage forms for oral administration include pharma- ceutically acceptable emulsions, solutions, suspensions, syrups and elixirs.Besides the peptide or nanoparticles thereof and the agent disclosed herein, liquid dosage forms may contain inert diluents, solubilizers and emulsifiers commonly used in the art, such as water or other solvents, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils, particularly cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil, glycerol, tetrahydrofurfuryl alcohol, polyethylene glycol and fatty acid esters of sorbitan, or mixtures of these substances.

[0079] In certain embodiments, a manufacturing process is contemplated in which the two components, the peptides disclosed herein or nanoparticles thereof, and the agent and pharmaceutical carrier disclosed herein, are already provided in a combined dry form ready to be reconstituted together. In other embodiments, it is contemplated to mix the peptides disclosed herein or nanoparticles thereof, any agent disclosed herein, and a pharmaceutical carrier to provide a pharmaceutical composition.

[0080] By simply adding a suitable pharma- ceutically acceptable diluent to the composition in the container, it is possible to provide the pharmaceutical composition in a one-step process. In certain embodiments, the container is preferably a syringe for administering the reconstituted pharmaceutical composition after contacting with the diluent. In certain embodiments, the peptides disclosed herein, or nanoparticles thereof, or drugs can be loaded into the syringe, which can then be closed with a stopper. The diluent is used in an amount to achieve the desired final concentration. The pharmaceutical composition may also include other useful components, such as ions, buffers, excipients, stabilizers, etc.

[0081] A "dry" pharmaceutical composition usually has only a residual moisture content, which can largely correspond to the moisture content of a comparable commercial product, for example, about 12% moisture as a dry product. Usually, a dry pharmaceutical composition according to the invention has a residual moisture content of preferably 10% or less, more preferably 5% or less, in particular 1% or less. The pharmaceutical composition can also have a lower moisture content, for example 0.1% or less. In certain embodiments, the pharmaceutical composition is provided in a dry state to prevent denaturation and allow storage stability.

[0082] The container may be any container suitable for containing (storing) the pharmaceutical composition, such as an inhaler, a syringe, a vial, a tube, etc. The pharmaceutical composition can then be applied via a specific needle of an actuation or syringe, or a suitable catheter. Common diluents include water for injection, NaCl (preferably 50-150 mM, in particular 110 mM), CaCl2 (preferably 10-80 mM, in particular 40 mM), sodium acetate (preferably 0-50 mM, in particular 20 mM) and mannitol (preferably up to 10% w / w, in particular 2% w / w). Preferably, the diluent may also contain a buffering agent or buffering system to buffer the pH of the reconstituted dry composition, preferably to pH 6.2-7.5, in particular pH 6.9-7.1.

[0083] In certain embodiments, the present disclosure contemplates a kit comprising a pharmaceutical composition as described herein, such as a container with a peptide, or nanoparticles thereof, or drug, as disclosed herein, and optionally a suitable diluent. Additional elements of the kit may be instructions for use, administration means, such as an inhaler, syringe, catheter, brush, etc. (if the composition is not already provided in the administration means), or other elements required for use in a medical (surgical) procedure, such as replacement needles or catheters, extra vials, or means for further covering the wound. In certain embodiments, the kit comprises a syringe containing the dry and stable hemostatic composition, and a syringe containing a diluent (or provided for removing the diluent from a separate diluent container).

[0084] In certain embodiments, the diluent is provided in a separate container. The container may preferably be a syringe. The diluent in the syringe can then be easily applied to the container to reconstitute the dry composition. If the container is also a syringe, both syringes can be completed together in a pack. Therefore, it is preferred to provide the dry composition in a syringe, which is completed with a diluent syringe together with a pharma- ceutically acceptable diluent to reconstitute this dry stable composition.

[0085] It is contemplated that any of the peptides disclosed herein may be modified with carbohydrate or polyethylene glycol groups to improve properties such as solubility, bioavailability, and / or biological degradation.

[0086] In certain embodiments, the present disclosure relates to a method of conjugating a peptide disclosed herein to a nanoparticle. In certain embodiments, the nanoparticle is composed of poloxamer-stabilized polypropylene sulfide. In certain embodiments, the nanoparticle has a diameter of 10-100 nm. In certain embodiments, the nanoparticle has a diameter of 20-50 nm, preferably 30 nm.

[0087] In certain embodiments, the present disclosure contemplates the use of the particles disclosed herein where the peptide sequence is attached to the nanoparticle and includes a peptide disclosed herein plus a C-terminal linker peptide, GGGGSC (SEQ ID NO:15). In certain embodiments, the particle includes a peptide sequence, for example, KPRRPYTDNYTRLRKQMAVKKYLNLILNGGGGSC (SEQ ID NO:12). In certain embodiments, the chemical bond between the peptide disclosed herein and the nanoparticle is a disulfide bond.

[0088] In certain embodiments, the pharma- ceutically acceptable excipient is an aerosolizing agent or a phospholipid. In certain embodiments, the aerosolizing agent is a hydrofluoroalkane, 1,1,1,2-tetrafluoroethane, 1,1,1,2,3,3,3-heptafluoropropane, propane, n-butane, isobutene, carbon dioxide, air, nitrogen, nitrous oxide, dimethyl ether, trans-1,3,3,3-tetrafluoroprop-1-ene, or a combination thereof. In certain embodiments, the phospholipid is dipalmitoyl phosphatidylcholine, palmitoyl-oleoyl phosphatidylglycerin, phosphatidylglycerin, or a combination thereof.

[0089] In certain embodiments, the pharmaceutical composition may be stored in a nebulizer, inhaler, or other container, optionally sealed, under pressure to spray the medication(s). The container may include a manually actuated or pressurized nebulizer device. Metered dose inhalers (MDIs) typically have a handheld aerosol canister that, when pressed, releases an amount of medication to be inhaled. Dry powder inhalers (DPIs) do not use a propellant to release the medication. Instead, a dry powder form of the peptide or nanoparticles thereof or drug is drawn into the lungs after breathing. In certain embodiments, a container containing the peptide or nanoparticles thereof disclosed herein is inserted into the device. Pressing a button or section of the device punctures the container. The powder contained in the container can be breathed through the mouthpiece of the device.

[0090] In certain embodiments, the pharmaceutical composition may comprise a naturally occurring or non-natural pulmonary surfactant composition. Contemplated natural pulmonary surfactant compositions typically comprise 70-90% phospholipids (PC), such as dipalmitoylphosphatidylcholine (DPPC), phosphatidylcholine, and phosphatidylglycerol (PG), 1-10% surfactant-associated proteins, apolipoproteins SP-A (SFTPA1), B (SFTPB), C (SFTPC), and D (SFTPD) (SP is an abbreviation for "surfactant-associated protein"), and 1-10% cholesterol (neutral fat). Artificial lung surfactants include colfosseryl palmitate, a mixture of DPPC with hexadecanol and tyloxapol added as spreading agents, Pumactant (Artificial Lung Expanding Compound or ALEC), a mixture of DPPC and PG, KL-4, which consists of DPPC, palmitoyl-oleoylphosphatidylglycerol, and palmitic acid combined with a 21 amino acid synthetic peptide that mimics the structural features of SP-B; a composition consisting of DPPC, PG, palmitic acid, and recombinant SP-C shares a nearly identical sequence to human SP-C, except that the palmitoylated cysteines are absent and replaced with phenylalanines to eliminate protein oligomerization. Contemplated animal-derived surfactants include beractant (Alveoafct™), extracted from bovine bronchoalveolar lavage fluid (Survanta™), extracted from minced bovine lung with added DPPC, palmitic acid and tripalmitin, calfactant (Infasurf™), extracted from calf bronchoalveolar lavage fluid, and poractant alfa (Curosurf™), extracted from material derived from minced porcine lung.

[0091] In certain embodiments, the pharmaceutical compositions disclosed herein may further comprise a glucocorticoid receptor agonist, such as triamcinolone, triamcinolone acetonide, prednisone, mometasone furoate, loteprednol etabonate, fluticasone propionate, fluticasone furoate, fluocinolone acetonide, dexamethasone cipecylate, decisobutyryl ciclesonide, clobetasol propionate, ciclesonide, budesonide, beclomethasone propionate, alclomethasone propionate; a p38 antagonist, such as losmapimod; a phosphodiesterase (PDE) inhibitor, such as methylxanthines, theophylline, and aminophylline. Respiratory medications include those selected from: selective PDE isoenzyme inhibitors, PDE4 inhibitors and PDE4D isoforms such as tetomilast, roflumilast, oglemilast, ibudilast; modulators of chemokine receptor function such as maraviroc, cenicriviroc, navarixin; leukotriene biosynthesis inhibitors, 5-lipoxygenase (5-LO) inhibitors, and 5-lipoxygenase-activating protein (FLAP) antagonists such as TA270 (4-hydroxy-1-methyl-3-octyloxy-7-sinapinoylamino-2(1H)-quinolinone), such as licofelone, zileuton, zafirlukast, or montelukast; and myeloperoxidase antagonists such as resveratrol and piceatannol.

[0092] In certain embodiments, the present disclosure relates to an in vitro cell culture composition comprising a basal medium and a peptide or nanoparticle thereof disclosed herein.

[0093] How to use In certain embodiments, the present disclosure relates to a peptide disclosed herein, e.g., Ant-8, KPRRPYTDNYTRLRKQMAVKKYLNLILN (SEQ ID NO:10), for use in a method of cancer treatment. In certain embodiments, the present disclosure contemplates a method of cancer treatment comprising administering an effective amount of a peptide disclosed herein, optionally in combination with a chemotherapeutic agent. In certain embodiments, the present disclosure relates to a method of treating or increasing an immune response against cancer in a subject in need thereof using a peptide disclosed herein, optionally conjugated to a nanoparticle, in combination with an antibody to an immune checkpoint molecule.

[0094] In certain embodiments, the subject has been diagnosed with a cancer selected from the group of melanoma, lung cancer, renal cancer, leukemia, non-Hodgkin's lymphoma, Hodgkin's lymphoma, myeloma, bladder cancer, pancreatic cancer, gastric cancer, esophageal cancer, glioblastoma, colon cancer, breast cancer, and prostate cancer.

[0095] In certain embodiments, the antibody against an immune checkpoint molecule is selected from the group of anti-PD1 antibodies including pembrolizumab (Keytruda™) and nivolumab (Opdivo™). In certain embodiments, the antibody against an immune checkpoint molecule is selected from the group of anti-PDL1 antibodies including atezolizumab (Tecentriq™), avelumab (Bavencio), durvalumab (Imfinzi™). In certain embodiments, the antibody against an immune checkpoint molecule is ipilimumab (Yervoy™).

[0096] In certain embodiments, the peptide therapeutic is administered by intravenous or subcutaneous injection. In certain embodiments, the peptide therapeutic is administered by inhaling the delivery agent into the alveoli. In certain embodiments, the peptide therapeutic is administered in a handheld delivery device powered by compressed gas. In certain embodiments, the peptide therapeutic is dissolved in a solution of sterile saline and administered as an aerosol.

[0097] In certain embodiments, it is contemplated that the peptides disclosed herein are used in certain cellular immunotherapies that are effective in the treatment of cancer, such as lymphocyte infusion or allogeneic bone marrow transplantation. Donor immune cells, particularly NK and T cells, have anti-cancer cytotoxic activity. VIP antagonism of the peptides increases the cellular immune response in vivo. VIP antagonism increases the cytotoxic activity of antigen-specific T cells and NK cells. VIP antagonism is expected to increase the anti-cancer activity of NK cells or antigen-specific T cells. VIP antagonism in conjunction with cellular immunotherapy is expected to increase the efficacy of the therapy. The absence of VIP is not expected to increase the "off-target" graft-versus-host disease activity of donor lymphocytes in recipients of allogeneic bone marrow transplantation. Thus, administering a VIP antagonist to a subject with cancer undergoing cellular therapy, such as donor lymphocyte infusion or allogeneic bone marrow transplantation, increases the anti-cancer activity of the therapy.

[0098] In certain embodiments, the present disclosure relates to a method of increasing an immune response to cell therapy, comprising administering to a subject a peptide disclosed herein in combination with cells. In certain embodiments, the subject has been diagnosed with leukemia or lymphoma. In certain embodiments, the cells are blood cells, bone marrow cells, white blood cells, T cells, natural killer cells, hematopoietic stem cells, G-CSF mobilized or non-mobilized blood mononuclear cells.

[0099] In certain embodiments, the cells are selected from the group consisting of autologous T cells, allogeneic cells from an HLA-matched donor, or allogeneic cells from an HLA-mismatched donor. In certain embodiments, the cells are bone marrow cells. In certain embodiments, the cells are blood mononuclear cells that contain / express granulocyte colony stimulating factor. Cell therapy may be performed with non-mobilized blood mononuclear cells.

[0100] In certain embodiments, it is contemplated that the peptides disclosed herein can be administered to a subject before, during, or after cell-based immunotherapy, including to a recipient or donor. The immunotherapy may be performed in combination with chemotherapy and / or radiation therapy. It is contemplated that the peptides may be used in combination with other immune stimulants, including, but not limited to, CpG oligonucleotides, granulocyte colony stimulating factor, granulocyte-macrophage colony stimulating factor, interferon alpha, pegylated interferon, interleukin-12, interleukin-2, and pegfilgrastim.

[0101] In certain embodiments, the present disclosure relates to a method of treating or preventing graft-versus-host disease in a subject, comprising administering an effective amount of a peptide disclosed herein to a subject following hematopoietic stem cell transplantation, or to a subject receiving or having been administered transplanted allogeneic tissue or cells. In certain embodiments, the subject has been administered transplanted allogeneic hematopoietic stem cells. In certain embodiments, the subject has been administered allogeneic hematopoietic stem cells isolated from peripheral blood. In certain embodiments, the subject has undergone chemotherapy followed by radiation therapy prior to receiving allogeneic hematopoietic stem cells.

[0102] In certain embodiments, the present disclosure relates to a method of treating cancer by performing stem cell transplantation, comprising administering to a subject a peptide disclosed herein in combination with a transplant of pluripotent hematopoietic stem cells derived from the subject (autologous) or a donor. Stem cells can be harvested from peripheral blood, such as from umbilical cord blood or placenta-derived stem cells or bone marrow. To limit the risk of rejection of transplanted stem cells or severe graft-versus-host disease, the donor usually has substantially the same human leukocyte antigens (HLA) as the recipient, although the donor may have incompatibility for certain antigens.

[0103] In certain embodiments, the present disclosure relates to methods of providing lymphocyte infusion following hematopoietic progenitor cell transplantation to treat hematologic malignancies (e.g., cancers of the blood or bone marrow, such as leukemia or lymphoma). The recipient of the transplant is typically infused with lymphocytes obtained in a leukapheresis from the original allogeneic stem cell (hematopoietic progenitor cell) donor.

[0104] In certain embodiments, the present disclosure relates to the extraction of lymphocytes from blood and their in vitro proliferation against tumor antigen(s) and optional exposure of the cells with appropriate stimulatory cytokines and / or peptides disclosed herein.

[0105] In certain embodiments, the present disclosure relates to a method of enhancing localized immune therapy, comprising administering a peptide disclosed herein in combination with the provision of an immune enhancing cream, such as imiquimod, that contains an interferon-producing drug that induces T cell activation.

[0106] In certain embodiments, it is contemplated that the peptides disclosed herein can be used in combination with adoptive cell therapy. For example, T cells with natural reactivity to cancer are found to infiltrate the tumor of a subject. The tumor can be harvested and these tumor infiltrating lymphocytes (TILs) can be expanded or made more effective using interleukin-2 (IL-2), anti-CD3 and alloreactive feeders in vitro. These T cells can then be transferred to the subject along with administration of a VIP antagonist. Prior to reinfusion, lymphodepletion of the recipient is usually performed to remove regulatory T cells and normal endogenous lymphocytes that compete with the transferred cells. It is also contemplated that adoptive cell transfer of lymphocytes can be transduced with a vector encoding a T cell receptor (TCR) that recognizes a cancer antigen.

[0107] In certain embodiments, the present disclosure relates to a method for increasing T cell activation and ex vivo proliferation by co-incubating human T cells with nanoparticles comprising small molecule antagonists of VIP signaling. In certain embodiments, human T cells are activated with plate-bound anti-CD3 antibodies. In certain embodiments, human T cells are activated in a mixed lymphocyte reaction. In certain embodiments, human T cells are activated in vitro by co-incubation with tumor-associated antigens. In certain embodiments, tumor-associated antigens are presented on tumor microvesicles. In certain embodiments, activated human T cells are infused into a human patient with cancer.

[0108] In certain embodiments, the activated human T cells are infused into a human patient with cancer. In certain embodiments, the human patient with cancer has leukemia. In certain embodiments, the human patient with cancer has lymphoma. In certain embodiments, the human patient with cancer has multiple myeloma. In certain embodiments, the human patient with cancer has epithelial cancer. In certain embodiments, the human patient with cancer has lung cancer. In certain embodiments, the human patient with cancer has breast cancer. In certain embodiments, the human patient with cancer has colon cancer. In certain embodiments, the human patient with cancer has prostate cancer. In certain embodiments, the human patient with cancer has malignant melanoma. In certain embodiments, the human patient with cancer has brain cancer.

[0109] In certain embodiments, the disclosure relates to a method of treating a subject diagnosed with cancer, comprising administering to a subject in need thereof cells in combination with a peptide disclosed herein. In certain embodiments, the subject has been diagnosed with leukemia. In certain embodiments, the subject has been diagnosed with lymphoma. In certain embodiments, the cell is a blood mononuclear cell. In certain embodiments, the cell is a bone marrow cell. In certain embodiments, the cell is a white blood cell. In certain embodiments, the cell is a T cell. In certain embodiments, the cell is a natural killer cell. In certain embodiments, the cell is a hematopoietic stem cell. In certain embodiments, the cell is a G-CSF mobilized blood mononuclear cell. In certain embodiments, the cell is an HLA-matched or mismatched allogeneic cell. In certain embodiments, the cell is a syngeneic cell. In certain embodiments, the cell is an autologous cell. In certain embodiments, the peptide has a C-terminal amide and / or is optionally modified with a carbohydrate or polyethylene glycol group.

[0110] In certain embodiments, the present disclosure relates to a method of treating leukemia comprising administering to a subject a peptide disclosed herein in combination with hematopoietic stem cell transplantation. In certain embodiments, the present disclosure relates to a method comprising expanding lymphocytes in vitro, providing the expanded cells and exposing the expanded cells with a peptide disclosed herein.

[0111] In certain embodiments, lymphocytes are extracted from blood or obtained by leukapheresis, hi certain embodiments, the expanded cells are further exposed to a stimulatory cytokine or interferon.

[0112] In certain embodiments, the present disclosure relates to a method of increasing anti-cancer immune responses by injecting a peptide disclosed herein or a nanoparticle expressing a peptide disclosed herein. In certain embodiments, activated T cells are infused into a patient with chronic CMV infection. In certain embodiments, activated T cells are infused into a patient with chronic EBV infection. In certain embodiments, activated T cells are infused into a patient with chronic BK virus infection. In certain embodiments, activated T cells are infused into a patient with chronic adenovirus infection.

[0113] In certain embodiments, the present disclosure relates to compositions and methods for reversing T cell senescence by interrupting vasoactive intestinal peptide (VIP) signaling and / or inhibiting phosphatidylinositol-3-kinase (PI3 kinase) inhibitor signaling for use in managing cancer and chronic viral infections. In certain embodiments, the present disclosure contemplates compositions and methods for reversing T cell senescence by mixing T cells in vitro with peptides disclosed herein or nanoparticles comprising peptides disclosed herein that prevent VIP from interacting with the VIP receptor and / or adding PI3 kinase inhibitors. In certain embodiments, the present disclosure contemplates proliferation of senescent T cells by mixing with PI3 kinase inhibitors, nanoparticles or peptides disclosed herein, VIP degrading enzymes, and combinations thereof.

[0114] In certain embodiments, the present disclosure contemplates a method of stimulating isolated T cells or expanding senescent T cells by exposing the T cells in vitro to antibodies that bind CD3 and / or CD28 in combination with a PI3 kinase inhibitor, idelalisib, a peptide disclosed herein or a nanoparticle disclosed herein, VIP-degrading enzyme, and combinations thereof. In certain embodiments, the present disclosure contemplates the use of anti-CD3 and anti-CD28 antibodies or binding agents, optionally coupled to a solid substrate such as magnetic beads.

[0115] In certain embodiments, the present disclosure contemplates methods of expanding CD28 and / or CD27 negative T cells using the in vitro cell cultures disclosed herein to provide replicated T cells with increased expression of CD28 and / or CD27 compared to levels prior to replication.

[0116] In certain embodiments, the disclosure contemplates a method of expanding T cells, where before, during, or after expansion of the T cells, the T cells are mixed with a vector having a nucleic acid sequence encoding a chimeric antigen receptor, the chimeric antigen receptor comprising a cancer targeting sequence, a transmembrane domain, a T cell costimulatory molecule domain, and a signaling component of the T cell antigen receptor domain, under conditions such that the cells express the chimeric antigen receptor on the surface of the cells.

[0117] In certain embodiments, the present disclosure relates to an in vitro cell culture comprising a basal medium and T cells and a peptide disclosed herein or nanoparticles comprising a peptide disclosed herein and optionally further comprising anti-CD3 and anti-CD28 antibodies, optionally immobilized on a solid substrate such as beads, comprising a phosphatidylinositol-3-kinase inhibitor, a VIP degrading enzyme, and combinations thereof. In certain embodiments, the T cells are purified from bone marrow cells or blood cells, peripheral blood.

[0118] In certain embodiments, the phosphatidylinositol-3-kinase inhibitor is selected from idelalisib, wortmannin, demethoxyviridin, perifosine, bupallisib, duvelisib, copanlisib, and alpelisib. In certain embodiments, the phosphatidylinositol-3-kinase inhibitor is selected from idelalisib at a concentration of 0.001, 0.1, 1, 10, greater than 100 nM, or between 10 nM and 10 micromolar, or between 10 nM and 500 nM, or between 10 nM and 1 micromolar in the medium.

[0119] In certain embodiments, the medium comprises an enzyme that hydrolyzes VIP. In certain embodiments, the medium comprises a VIP-degrading enzyme, such as peptidase, serine peptidase, tryptase, chymase, or human chymase 1 (CMA1). In certain embodiments, the medium has at least 0.001, 0.01, 0.1, or 1 microgram of a VIP-degrading enzyme, such as mast cell chymase. In certain embodiments, the present disclosure contemplates a T cell medium comprising a basal medium and isolated cells expressing CD3 and / or CD4 and / or CD8, negative for CD27 and / or CD28, and a PI3 kinase inhibitor, a peptide disclosed herein or a nanoparticle comprising a peptide disclosed herein, and combinations thereof. The cells can be isolated by negative or positive selection using a binder attached to a solid support, such as beads, magnetic beads, or particles of a fluorescent binder.

[0120] In certain embodiments, the anti-CD3 antibody and the anti-CD28 antibody are immobilized on beads, magnetic beads or a solid surface. In certain embodiments, more than 5.0%, 10%, or 15% of the total cells of the culture express CD3 and / or CD4 and / or CD8. In certain embodiments, more than 20%, 25%, or 50% of the total cells express CD3 and / or CD4 and / or CD8. In certain embodiments, more than 15%, 20%, or 30% of the T cells of the culture are negative for CD28 and / or CD27. In certain embodiments, more than 20%, 25%, or 50% of the T cells are negative for CD28 and / or CD27.

[0121] In certain embodiments, purified T cells are obtained by centrifuging blood under conditions that separate plasma and red blood cells, providing purified T cells to a mixture of white blood cells between the plasma and red blood cells, hi certain embodiments, purified T cells are obtained by bone marrow aspirate or bone marrow biopsy.

[0122] In certain embodiments, purified T cells are obtained by mixing cells with a fluorescent marker that binds CD3 and purifying the cells by fluorescence activated cell sorting. In certain embodiments, purified T cells are obtained by mixing cells with a magnetic marker that binds CD3 and purifying the cells by magnetic sorting. In certain embodiments, purified T cells are obtained by mixing cells with a fluorescent marker that binds CD3 and / or CD4 and / or CD8 and purifying the cells by fluorescence activated cell sorting. In certain embodiments, purified T cells are obtained by mixing cells with a magnetic marker that binds CD3 and / or CD4 and / or CD8 and purifying the cells by magnetic sorting.

[0123] In certain embodiments, the present disclosure contemplates a solid substrate, such as beads, having anti-CD3 and anti-CD28 antibodies and having VIP degrading enzyme bound to the surface. In certain embodiments, the beads are arrayed in media and T cells are grown on top of the media such that the beads are intracellular. In a particular embodiment, the VIP degrading enzyme is human CMA1 Accession No. GenBank: AAI03975.1:

[0124] Contains MLLKLKEKASLTLAVGTLPFPSQFNFVPPGRMCRVAGWGRTGVLKPGSDTLQEVKLRLMDPQACSHFRDFDHNLQLCVGNPRKTKSAFKGDSGGPLLCAGVAQGIVSYGRSDAKPPAVFTRISHYRPWINQILQAN (SEQ ID NO:13).

[0125] In a particular embodiment, the VIP degrading enzyme is human recombinant enkephalinase (neutral endopeptidase EC 3.4.24.11) and has the following (SEQ ID NO:14): .

[0126] In certain embodiments, the cell cultures and methods described herein further comprise IL-12. In certain embodiments, it is contemplated that IL-12 enhances the effect of the peptides disclosed herein or nanoparticles thereof on the proliferation of T cells stimulated in vitro with antibodies against CD3 and CD28.

[0127] In certain embodiments, the present disclosure relates to expanding T cells with naturally occurring reactivity against cancer that may be found infiltrating a tumor in a subject, or expanding or reversing T cell senescence. The tumor can be harvested and these tumor infiltrating lymphocytes (TILs) can be expanded using the methods disclosed herein.

[0128] Some cancers are caused by viruses against which traditional vaccines, such as HPV vaccines and Hepatitis B, prevent these cancers. It is contemplated that the peptides disclosed herein can be administered in combination with these vaccines to improve therapeutic efficacy.

[0129] It is believed that cancer cells develop and are destroyed by the immune system, and cancer is formed when the immune system is unable to destroy the cancer cells. One approach to cancer vaccination is to isolate proteins from cancer cells, immunize cancer patients against these proteins, and stimulate an immune response that kills the cancer cells. Cancer vaccines are contemplated for the treatment of breast, lung, colon, skin, kidney, prostate and other cancers. In certain embodiments, the present disclosure relates to cancer treatment by administering the peptides disclosed herein in combination with cancer antigens.

[0130] In certain embodiments, the present disclosure relates to a method for treating or preventing a viral infection comprising administering a peptide disclosed herein to a subject diagnosed as being at risk for, exhibiting symptoms of, a viral infection. In certain embodiments, the subject is immunocompromised, or the subject is an allogeneic bone marrow transplant donor or recipient. In typical embodiments, the subject is an organ transplant recipient, undergoing hemodialysis, diagnosed with cancer, receiving immunosuppressants, and / or diagnosed with HIV infection. In certain embodiments, the present disclosure relates to preventing a viral infection in an immunocompromised subject at risk for infection by administering a peptide disclosed herein and, optionally, one or more antiviral drugs.

[0131] In some embodiments, the disclosure relates to the use of the peptides disclosed herein in the manufacture of antiviral medicines for the treatment of viral infection. In some embodiments, the subject has been diagnosed with a chronic viral infection. In certain embodiments, the subject is undergoing serological monitoring. In some embodiments, the administration is performed under conditions where no further viral infection is detected. In some embodiments, the subject has been diagnosed with an RNA virus, a DNA virus, or a retrovirus. In some embodiments, the subject has been diagnosed with a double-stranded DNA virus, a sense single-stranded DNA virus, a double-stranded RNA virus, a sense single-stranded RNA virus, an antisense single-stranded RNA virus, a sense single-stranded RNA retrovirus, or a double-stranded DNA retrovirus. In some embodiments, the subject has been diagnosed with a rotavirus, influenza virus, herpes virus, hepatitis virus, or lentivirus. In some embodiments, the subject's viral titer is reduced after treatment compared to before treatment.

[0132] In some embodiments, the subject is infected with influenza A virus, including subtype H1N1, influenza B virus, influenza C virus, rotavirus A, rotavirus B, rotavirus C, rotavirus D, rotavirus E, SARS coronavirus, human adenovirus types (HAdV-1-55), human papillomavirus (HPV) types 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, parvovirus B19, molluscum contagiosum virus, JC virus (JCV), BK virus, Merkel cell polyomavirus, Coxsackie A virus, norovirus, rubella virus, lymphocytic choriomeningitis virus (LCMV), yellow fever virus, measles virus, mumps virus, respiratory syncytial virus, The following viruses have been diagnosed: Rinderpest virus, California encephalitis virus, hantavirus, rabies virus, Ebola virus, Marburg virus, herpes simplex virus type 1 (HSV-1), herpes simplex virus type 2 (HSV-2), varicella zoster virus (VZV), Epstein-Barr virus (EBV), cytomegalovirus (CMV), herpes lymphotropic virus, roseolovirus, Kaposi's sarcoma-associated herpesvirus, hepatitis A (HAV), hepatitis B (HBV), hepatitis C (HCV), hepatitis D (HDV), hepatitis E (HEV), human immunodeficiency virus (HIV), human T-lymphotropic virus type 1 (HTLV-1), spleen focus-forming virus (SFFV), or xenotropic murine leukemia virus-related virus (XMRV).

[0133] In some embodiments, the disclosure relates to the treatment or prevention of viral infection by administering a peptide disclosed herein in combination with a second antiviral drug. In further embodiments, the subject is treated with abacavir, acyclovir, acyclovir, adefovir, amantadine, amprenavir, ampligen, arbidol, atazanavir, atripla, boceprevir, cidofovir, combivir, darunavir, delavirdine, didanosine, docosanol, edoxudine, efavirenz, emtricitabine, enfuvirtide, entecavir, famciclovir, fosamprenavir, foscarnet, ganciclovir, idoxuridine, imiquimod, indinavir, inosine, interferon type III, interferon type II, interferon type I, lamivudine, lopinavir. , maraviroc, moroxydine, methisazone, nelfinavir, nevirapine, oseltamivir (Tamiflu), peginterferon alfa-2a, penciclovir, peramivir, pleconaril, podophyllotoxin, raltegravir, ribavirin, rimantadine, ritonavir, saquinavir, stavudine, tenofovir, tenofovir disoproxil, tipranavir, trifluridine, trizivir, tromantadine, Truvada™ (emtricitabine / tenofovir disoproxil fumarate), valacyclovir, valacyclovir, vicriviroc, vidarabine, viramidine, zalcitabine, zanamivir, and / or zidovudine. In certain embodiments, the subject is administered a pharmaceutical composition comprising a peptide disclosed herein and a second antiviral agent.

[0134] In certain embodiments, the present disclosure relates to the treatment of subjects with post-infection viral infections by administering the peptides and immunoglobulins disclosed herein.

[0135] In certain embodiments, the present disclosure relates to the treatment or prevention of viral infections by administering the peptides disclosed herein and a viral vaccine, or in the absence of a viral vaccine.

[0136] In certain embodiments, the present disclosure relates to enhancing an immune response to a vaccine comprising administering a peptide disclosed herein to a subject in need thereof. Typically, the vaccine is a vaccine, such as herpes zoster vaccine, smallpox vaccine, polio vaccine, pertussis vaccine, influenza vaccine, diphtheria vaccine, tetanus vaccine, meningococcal vaccine, influenza A vaccine including subtype H1N1 vaccine, influenza B vaccine, influenza C vaccine, rotavirus A vaccine, rotavirus B vaccine, rotavirus C vaccine, rotavirus D vaccine, rotavirus E vaccine, SARS coronavirus vaccine, human adenovirus (HAdV-1-55) vaccine, human papillomavirus (HPV) vaccine, parvovirus B19 vaccine, molluscum contagiosum vaccine, JC vaccine, BK vaccine, Merkel cell polyomavirus vaccine, coxsackie A vaccine, norovirus vaccine, rubella vaccine, lymphocytic choriomeningitis vaccine, yellow fever vaccine, measles vaccine, mumps vaccine, respiratory syncytial vaccine, or the like. In one embodiment, the vaccine is selected from the group consisting of rabies vaccine, ... In certain embodiments, the vaccine is for subjects who have been diagnosed with a chronic viral infection.

[0137] In certain embodiments, the vaccine comprises a protein or peptide, a carbohydrate, a sugar, a polysaccharide, or a nucleic acid. Typically, the vaccine is an attenuated, replication-competent or inactivated virus. In certain embodiments, the vaccine comprises live, killed or inactivated prokaryotic or eukaryotic cells.

[0138] In certain embodiments, human T cells are activated in vitro by co-incubation with viral antigens. In certain embodiments, viral antigens are presented on microvesicles. In certain embodiments, viral antigens are presented on dendritic cells.

[0139] Nucleic acid vaccines, usually DNA plasmids, are generally designed to encode and / or produce one or more antigens from a pathogen. The nucleic acid is transfected or infects a host cell, and the cell's internal machinery expresses proteins. These proteins are recognized as foreign and therefore, when processed and displayed on the surface by the host cell, trigger an immune response. Cytotoxic T lymphocyte responses can also be boosted by co-vaccination with costimulatory molecules such as GM-CSF, B7-1, or B7-2. In certain embodiments, the peptides disclosed herein can be administered in combination with a nucleic acid vaccine or other costimulatory molecules.

[0140] In certain embodiments, the present disclosure relates to vaccine compositions comprising the peptides disclosed herein and methods of administering the peptides disclosed herein in combination with a vaccine. In certain embodiments, the vaccine comprises an antigen from a pathogen, presented to the immune system from a weakened or killed form of the microorganism or its toxin. The antigen stimulates the immune system. Vaccines may be prophylactic (e.g., to prevent or reduce the effects of future infection with any pathogen) or therapeutic by administration after diagnosis of infection or disease.

[0141] Some vaccines contain killed but previously toxic microorganisms that have been destroyed with chemicals or heat. Influenza vaccines, cholera vaccines, bubonic plague vaccines, polio vaccines, Hepatitis A vaccines, and rabies vaccines are examples of killed vaccines contemplated by the present disclosure.

[0142] Some vaccines contain live attenuated microorganisms, usually live viruses cultivated under conditions that neutralize their specific toxicity, or use closely related but less dangerous organisms to generate a broad immune response, but which may be bacterial in nature.

[0143] In certain embodiments, the vaccine is a protein subunit. Rather than introducing an inactivated or attenuated microorganism into the immune system, fragments thereof can be used to generate an immune response. Examples include subunit vaccines against Hepatitis B virus consisting of only the surface proteins of the virus, virus-like particle (VLP) vaccines against human papillomavirus (HPV) consisting of the major capsid proteins of the virus, and hemagglutinin and neuraminidase subunits of the influenza virus.

[0144] In certain embodiments, the vaccine comprises a polysaccharide. Certain bacteria have polysaccharide coats that are normally immunogenic. By conjugating these polysaccharides to proteins (e.g., toxins), the immune system can be directed to recognize the polysaccharide as if it were a protein antigen.

[0145] Toxoid vaccines are made from inactivated toxic compounds. Examples of toxoid-based vaccines include diphtheria and tetanus toxoids. In certain embodiments, the peptides disclosed herein are administered in combination with DPT. DPT (also DTP and DTwP) refers to a class of combination vaccines against three infectious diseases in humans: diphtheria, pertussis (whooping cough) and tetanus. Vaccine components include diphtheria and tetanus toxoids and whole cells (wP) of the killed organism that causes whooping cough. DTaP (also known as Tdap, DTPa, and TDaP) refers to a similar combination vaccine in which the pertussis component is acellular. DT or TD vaccines lacking the pertussis component are also contemplated.

[0146] Other specific vaccines contemplated by the present disclosure include anthrax vaccines, e.g., culture filtrate of the non-virulent, non-encapsulated strain known as V770-NP1-R, Bacille Calmette-Guerin (BCG), e.g., a live attenuated strain of Mycobacterium bovis, Haemophilus influenzae B vaccines, e.g., a Hib polysaccharide-protein conjugate vaccine, Hepatitis A vaccines, e.g., inactivated Hepatitis A virus, Hepatitis B vaccines, e.g., Hepatitis B surface antigen, Human Papillomavirus (HPV) vaccines, e.g., non-infectious virus-like particles constructed from the L1 proteins of HPV types 6, 11, 16 and 18, and meningococcal vaccines, e.g., meningococcal subgroups A, C, Y, and W-135 strains individually bound to the diphtheria toxoid protein.

[0147] In certain embodiments, the disclosure relates to a method of treating an active cytomegalovirus infection comprising administering an effective amount of a vasoactive intestinal peptide antagonist disclosed herein to a subject diagnosed with, and exhibiting signs or symptoms of, an active cytomegalovirus infection, wherein the vasoactive intestinal peptide antagonist comprises a peptide having a C-terminal amide, optionally modified with a carbohydrate or polyethylene glycol group.

[0148] In certain embodiments, the subject has a compromised immune system, hi certain embodiments, the subject is a transplant recipient.

[0149] In certain embodiments, the present disclosure relates to a method of reducing an active cytomegalovirus infection comprising administering to a subject suffering from an active cytomegalovirus infection an effective amount of a vasoactive intestinal peptide antagonist disclosed herein, the vasoactive intestinal peptide antagonist comprising a peptide having a C-terminal amide, optionally modified with a carbohydrate or polyethylene glycol group, In certain embodiments, the subject's cytomegalovirus titer is reduced after administration of the vasoactive intestinal peptide antagonist compared to before treatment. EXAMPLES

[0150] Improved VIP Antagonists We evaluated whether tumor-specific expression of vasoactive intestinal polypeptide antagonists represents a mechanism of tumor-mediated immune escape. A spectrum of VIP expression in tumors with the highest expression is seen in pancreatic exocrine carcinomas and the lowest expression in melanomas. In general, the level of VIP expression by tumors is inversely proportional to the expression of other co-inhibitory pathway molecules such as PDL1. Tumors that express and secrete VIP may have mutations in the VIP coding sequence, and the resulting peptide molecule has improved pharmacokinetics or pharmacodynamics in the tumor microenvironment. Pharmacokinetic advantages may be the result of mutations that reduce VIP's low susceptibility to proteases, improving its half-life. Pharmacodynamic advantages may be the result of mutations that increase binding affinity for the receptor, thus enhancing signal transduction.

[0151] Experiments were performed to determine whether mutant VIP produced by tumors leads to more sustained suppression of anticancer T cells in the tumor microenvironment. Analysis of mutations in selected genes from accumulated tumor sequences revealed that multiple cancers have mutations in the coding sequence of VIP. In particular, there were 140 missense mutations and 17 truncation mutations within the VIP gene cluster listed in the Cancer Genome Atlas. Mutations were identified in the coding sequence of the 28 amino acid VIP peptide present in breast cancer, prostate adenocarcinoma, esophageal adenocarcinoma, cutaneous melanoma, small cell lung cancer, gastric adenocarcinoma, endometrial cancer, cutaneous melanoma, esophageal adenocarcinoma, colorectal adenocarcinoma, uterine cancer, hepatocellular adenoma, lung adenocarcinoma, and gastric adenocarcinoma. Eight specific mutations were present in the C-terminal amino acid, which includes the alpha helix of VIP that binds to the receptor.

[0152] The α-helical sequence is common in VIP, a peptide agonist, and is immunosuppressive. VIPhyb (SEQ ID NO:1) is an antagonistic peptide that differs by six internal amino acids from native VIP (SEQ ID NO:2) (see FIG. 1). Eight sequences, ANT-1-8, were prepared in the presence of the six N-terminal amino acids of VIPhyb (see SEQ ID NOs:3-10 in FIG. 1). In particular, the ANT-1 sequence has an amino acid substitution T-A at amino acid position 7; the ANT-2 sequence has an amino acid substitution D-V at amino acid position 8; ANT-3 has an amino acid substitution Y-C at amino acid position 10; ANT-4 has an amino acid substitution R-S at amino acid position 12; ANT-5 has an amino acid substitution M-I at amino acid position 17; and ANT-6 has an amino acid substitution K-N at amino acid position 20. ANT-7 has an amino acid substitution LM at amino acid position number 23. ANT-8 has an amino acid substitution S L at amino acid position number 25. Each of the alternative antagonistic peptide sequences ANT-1 through ANT-8 were synthesized.

[0153] Peptides (SEQ ID NOs:3-10 in FIG. 1) were added to the short-term medium of luciferase-positive T cells cultured in the presence of plate bound anti-CD3 antibody in 96-well plates. The concentration of anti-CD3 antibody was 0.5 mcg / ml or 1 mcg / ml. T cells from luciferase transgenic mice were added in the presence of low amounts of IL-2 and 0.5 micromolar, 1 micromolar, or 3 micromolar concentrations of the original VIPhyb peptides, or alternative ANT-1-ANT-8 peptide sequences. FIG. 2 shows T cell proliferation at 24 hours in the presence of ANT-1-ANT-8 at 1 micromolar concentration. FIG. 3 shows increased T cell proliferation at 24 hours in the presence of ANT-8 peptides at 0.5 micromolar, 1 micromolar, or 3 micromolar concentrations compared to the original VIPhyb peptides at the same corresponding concentrations.

[0154] Increased T cell proliferation using lower concentrations of anti-CD3 with ANT-8 peptide was more modest and not significantly different from control media containing plate bound anti-CD3 antibody without added peptide, resulting in only a slight increase over control media.A unique peptide sequence was identified, ANT-8 (SEQ ID NO:10), that has improved antagonist activity compared to native VIPhyb.

Claims

1. A peptide comprising: KPRRPYTDNYTRLRKQMAVKKYLNLILN (SEQ ID NO: 10).

2. 2. The peptide of claim 1, wherein an amino, carboxyl, hydroxyl, or thiol group is substituted in the peptide.

3. The peptide of claim 1 , wherein the peptide is attached to a nanoparticle.

4. A pharmaceutical composition comprising the peptide of claim 1 and a pharma- ceutically acceptable excipient.

5. 5. The pharmaceutical composition of claim 4 in the form of a capsule, tablet, pill, powder, or granules.

6. 5. The pharmaceutical composition of claim 4 in the form of a sterile pH buffered aqueous saline solution.

7. 7. The pharmaceutical composition of claim 6, in the form of a container configured to spray a liquid or a sealed container having a propellant.

8. A nucleic acid encoding the peptide of claim 1 or 2 operably combined with a promoter.

9. A recombinant vector comprising the nucleic acid of claim 8.

10. A cell comprising the recombinant vector of claim 9.

11. A method for increasing the activity and proliferation of T cells in vitro or ex vivo, comprising mixing T cells with a peptide according to claim 1 or 2.

12. The method of claim 11, wherein the admixture of T cells is in combination with an anti-CD3 antibody and / or an anti-CD28 antibody.

13. The method of claim 11, wherein the admixture of T cells is combined with a phosphatidylinositol 3-kinase delta (PI3Kδ) inhibitor.

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