JP7307500B2 - IL-2 receptor binding compounds - Google Patents

Description

This application is filed under 35 U.S.C. S. C. U.S. Provisional Application No. 62/785,754, filed Dec. 28, 2018, and U.S. Provisional Application No. 62/715,097, filed Aug. 6, 2018, pursuant to §119(e) , each of which is incorporated by reference in its entirety.

The present disclosure relates to IL-2Rβ ligands, IL-2Rγc ligands, and compounds with IL-2Rβ and/or IL-2Rγc ligands. Compounds such as synthetic heterodimers and recombinant fusion proteins comprising IL-2Rβ and/or IL-2Rγc ligands can be IL-2 receptor agonists.

SEQUENCE LISTING This application contains a Sequence Listing which has been submitted electronically in ASCII format and is hereby incorporated by reference in its entirety. The above ASCII copy made on July 31, 2019 is 62AJ-000120PC-297328_SL. txt and is 328,997 bytes in size.

Recombinant human interleukin-2 (IL-2) was one of the first immuno-oncology agents studied in the clinic and was used in several particularly difficult cancers, melanoma and renal cancer, in the 1990s. Approved by the FDA for use against Although IL-2 is effective and produces durable responses in up to 10% of patients with these tumors, its usefulness is limited by very severe dose-limiting toxicities. In addition, the efficacy of IL-2 to induce T cell-mediated anti-tumor responses is compromised by concurrent IL-2-driven upregulation of the T cell suppressive system. Strategies to reduce the toxicity of IL-2 therapy and circumvent the immunosuppressive limitations on its anti-tumor activity have been continuously explored. To date, moderately effective strategies have been developed to control systemic exposure, and thus toxicity, of this potent biologic. Unraveling the complex biology of IL-2 has led to modifications of the native IL-2 molecule to alter the balance between tumor toxicity and suppression. However, these approaches are limited by the use of native IL-2 as a template, thus retaining the undesirable structure-driven bioactivity component of the parent molecule.

IL-2 is important for its anti-tumor properties and exerts potent stimulatory effects on NK and cytotoxic CD8+ T cells. However, anti-tumor effects are paradoxically suppressed by IL-2-directed stimulation of T regulatory cells (Tregs), which effectively dampens anti-tumor immune responses. This dual effect of IL-2 is largely controlled by the properties of the IL-2 receptor (IL-2R) subunits expressed on various cells responsible for immune homeostasis. IL-2 is recognized by a combination of three receptor subunits that are differentially and conditionally expressed on many types of immune cells. Two signaling subunits, known as IL-2Rβ (β) and IL-2Rγ common (γc), initiate signaling when binding IL-2 brings them into juxtaposition in the correct orientation. IL-2 binds IL-2Rβγc with an affinity of approximately 1 nM to form an active ternary complex. Most immune cells express IL-2Rβ and IL-2Rγc subunits at varying levels. There is also a third non-signaling IL-2R subunit, IL-2Rα (also known as CD25), expressed on a subset of immune cells, particularly Tregs. The complex of IL-2Rαβγc has a very high affinity for IL-2 (approximately 10 pM), thus cells expressing all three subunits are much more sensitive to IL-2. is high. A common and well-supported strategy for improving the efficacy of IL-2 receptor agonists against tumors is to manipulate IL-2R selectivity to reduce IL-2 binding to the IL-2Rα subunit. It involves reducing but maintaining IL-2Rβγc binding and signaling to favor the infiltration and stimulation of cytotoxic effector T cells (Teff cells) over Tregs at the tumor site.

The cause of IL-2 toxicity in the clinical setting is poorly understood, but is thought to be the result of exaggerated peripheral immune stimulation of IL-2Rβγc-expressing T cells with excessive release of inflammatory cytokines. Toxicity is induced by frequent administration of high doses of IL-2 required to maintain adequate tumor exposure due to the short half-life of natural cytokines.

Strategies to address the limitations of IL-2 as a useful immuno-oncology therapy utilize mutants, fusion proteins, or chemically modified IL-2 to alter the complex biology of immunomodulators. One example is a modified form of IL-2 that serves the dual purpose of altering receptor subunit binding specificity and extending the circulating half-life of reversibly inactive prodrugs of IL-2. decorated with two large cleavable polyethylene glycol (PEG) moieties. As the prodrug circulates systemically, the cascade of PEG removal confers a complex pharmacokinetic (PK) profile for the various active and inactive forms of the cytokine, leading to a low sustained to active IL-2 agonism. It provides peripheral exposure, thereby avoiding the C _max -driven severe side effects of high-dose IL-2. The last two PEGs that are cleaved are located near the IL-2Rα binding site and interfere with IL-2Rα binding, but allow IL-2Rβγc signaling and are therefore more cytotoxic than suppressive Treg activity. Favors T cell activity. This provides a promising therapeutic molecule that addresses two major deficiencies of IL-2 as an anticancer therapeutic: (a) avoiding activation of IL-2Rαβγc on Tregs, and ( b) Prolonged half-life of IL-2Rβγc activating compounds. However, these effects are necessarily intertwined and difficult to optimize separately, as is often required during preclinical and clinical development. This limits the use of bioactive IL-2 proteins as a starting point for conferring multiple new properties.

According to the present invention, IL-2Rβ ligands exhibit a binding affinity (IC50) for human IL-2Rβ subunits of less than 100 μM.

According to the present invention, the IL-2Rβ ligand has an amino acid sequence selected from any one of SEQ ID NOs: 1-193, SEQ ID NOs: 578-903, and SEQ ID NOs: 1028-1050. include.

According to the present invention, IL-2Rγc ligands exhibit a binding affinity (IC50) for human IL-2Rγc subunits of less than 100 μM.

According to the present invention, the IL-2Rγc ligand comprises an amino acid sequence selected from any one of SEQ ID NOs:194-267 and SEQ ID NOs:904-1027.

According to the invention, the compound comprises at least one of an IL-2Rβ ligand according to the invention and/or an IL-2Rγc ligand according to the invention.

According to the invention, a pharmaceutical composition comprises an IL-2Rβ ligand according to the invention and/or an IL-2Rγc ligand according to the invention, a compound according to the invention, or a combination thereof.

According to the invention, a method of treating cancer in a patient comprises administering to a patient in need of such treatment a therapeutically effective amount of a pharmaceutical composition according to the invention.

According to the invention, a method of treating an autoimmune disease in a patient comprises administering to a patient in need of such treatment a therapeutically effective amount of a pharmaceutical composition according to the invention.

In accordance with the present invention, methods are provided for screening compounds for IL-2Rβ and/or IL-2Rγc activity, methods for activating the human IL-2 receptor, the IL-2 receptor signaling pathway being responsible for disease pathogenesis. method of treating a disease in a patient associated with , a method of treating a disease in a patient wherein activation of the IL-2 receptor is effective in treating the disease, inhibition of the IL-2 receptor is effective in treating the disease Methods of treating diseases in patients where cells expressing IL-2Rβ and/or IL-2Rγc subunits are implicated in disease etiology, IL-2Rγc subunits and/or L-2Rγc subunits A method of treating a disease in a patient in which cells expressing the unit are implicated in the etiology of the disease, a method of treating a disease in a patient in which reducing the sensitivity of Treg cells to IL-2 is effective in treating the disease, IL- Methods of Imaging Cells Expressing 2Rβ Subunit and/or IL-2Rγc Subunit, Diagnosing Disease in Patients Where Disease is Associated with Cells Expressing IL-2Rβ Subunit and/or IL-2Rγc Subunit Methods, methods of targeting compounds to cells expressing IL-2Rβ subunit and/or IL-2Rγc subunit, and compounds cytotoxic to cells expressing IL-2Rβ subunit and/or IL-2Rγc subunit including methods of delivering

According to the present invention, a group of IL-2Rβ ligands having amino acid sequences of SEQ ID NOS: 1-163, 164-182, 578-808 and 1028-1042, wherein the IL-2Rβ subunit binding site is An IL-2Rβ ligand that competitively binds to the binding site with each of the other IL-2Rβ ligands in the group and has the amino acid sequence of SEQ ID NO: 1041 competes with the group of IL-2Rβ ligands for binding to the binding site. First, IL-2 is a binding site for IL-2Rβ subunits that does not compete with a group of IL-2Rβ ligands for binding to the binding site.

According to the invention, the binding site of the IL-2Rγc subunit comprising SEQ ID NOs: 194-210, 904-913, 211-233, 914-920, 234-245, 246-254, 921-922, 265- 267, and a group of IL-2Rγc ligands having amino acid sequences of 932-940 that competitively bind to the binding site with each of the other IL-2Rγc ligands within the group, resulting in an IL having the amino acid sequence of SEQ ID NO:948. - IL-2Rγc subunit binding, wherein the 2Rγc ligand does not compete with the group of IL-2Rγc ligands for binding to the binding site, and IL-2 does not compete with the group of IL-2Rγc ligands for binding to the binding site. It is a part.

Reference is now made to specific compounds and methods. The disclosed embodiments are not intended to limit the scope of the claims. On the contrary, the claims are intended to cover all alternatives, modifications and equivalents.

The drawings described herein are for illustration purposes only. The drawings are not intended to limit the scope of the disclosure.

STAT5 phosphorylation in NK-92 cells by IL-2Rβγc agonists according to the present disclosure by Western blot analysis. STAT5 phosphorylation in TF-1 cells by IL-2Rβγc agonists according to the present disclosure by Western blot analysis. RT-qPCR gene expression profiling comparing several transfected TF-1 cell populations is shown. Activation of STAT5, ERK1/2, and AKT in NK-92 cells by IL-2Rβγc agonists according to the present disclosure by Western blot analysis. STAT5 phosphorylation in NK-92 cells by IL-2Rβγc agonists according to the present disclosure by Western blot analysis. FIG. 4A shows dose-response curves of pSTAT5 in NK-92 cells with the IL-2Rβγc agonists evaluated in FIG. 4A. Results of NK-92 proliferation assays using IL-2Rβγc agonists according to the present disclosure (FIG. 5A), with rhIL-2 (FIG. 5B), and overlay comparisons (FIG. 5C) are shown. ELISA signals for different phage binding at pH 6.0 and pH 7.5 are shown. ELISA signals of phages showing pH-dependent binding compared to non-pH-dependent clones that showed similar binding affinities at pH 6.0 and pH 7.5 are shown. Phage binding _IC50 curves of pH-independent clones (Fig. 8A) and pH-dependent phages (Fig. 8B) at pH 6.0 and pH 7.5 are shown. IC ₅₀ curves for peptide binding to IL-2Rβ-Fc are shown for pH-independent clones (FIG. 9A) and peptides from the screen that showed pH-dependent binding (FIG. 9B). STAT5 phosphorylation in NK-92 cells by heterodimeric IL-2Rβγc agonists provided by the present disclosure.

A dash (“-”) that is not between two letters or symbols is used to indicate a point of attachment for a moiety or substituent. For example, -CONH ₂ is linked through a carbon atom and -X ¹ -X ² - indicates amino acids X ¹ and X ² covalently linked through a single bond.

"Alkyl" means a saturated or unsaturated, branched or straight chain, monovalent hydrocarbon derived by removing one hydrogen atom from a single carbon atom of a parent alkane, alkene, or alkyne refers to radicals. Examples of alkyl groups include ethyl such as methyl, ethanyl, ethenyl, and ethynyl, propan-1-yl, propan-2-yl, prop-1-en-1-yl, prop-1-en-2-yl , prop-2-en-1-yl (allyl), prop-1-yn-1-yl, prop-2-yn-1-yl and the like propyl, butan-1-yl, butan-2-yl, 2 -methylpropan-1-yl, 2-methylpropan-2-yl, but-1-en-1-yl, but-1-en-2-yl, 2-methyl-prop-1-en-1-yl , but-2-en-1-yl, but-2-en-2-yl, but-1,3-dien-1-yl, but-1,3-dien-2-yl, but-1-yne Butyls such as -1-yl, but-1-yn-3-yl, but-3-yn-1-yl and the like can be mentioned. The term “alkyl” includes groups having any degree or level of saturation, i.e., groups having exclusively carbon-carbon single bonds, groups having one or more carbon-carbon double bonds, groups having one or more carbon It is specifically intended to include groups with carbon-carbon triple bonds, as well as groups with combinations of carbon-carbon single, double, and triple bonds. Where specific saturation levels are intended, the terms alkanyl, alkenyl, and alkynyl are used. In certain embodiments, the alkyl group is C _1-6 alkyl, C _1-5 alkyl, C _1-4 alkyl, C _1-3 alkyl, and in certain embodiments ethyl or methyl.

"Cycloalkyl" refers to saturated or partially unsaturated cyclic alkyl radicals. In certain embodiments, the cycloalkyl group is C _3-6 cycloalkyl, C _3-5 cycloalkyl, C _5-6 cycloalkyl, cyclopropyl, cyclopentyl, and in certain embodiments cyclohexyl. . In certain embodiments, cycloalkyl is selected from cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

"Heterocycloalkyl" by itself or as part of another substituent, a saturated or unsaturated cyclic alkyl in which one or more carbon atoms (and certain attached hydrogen atoms) are independently replaced with the same or different heteroatoms refers to a radical or parent aromatic ring system in which one or more carbon atoms (and certain attached hydrogen atoms) are independently replaced with the same or different heteroatoms and the ring system violates the Hackel rule . Examples of heteroatom(s) to replace carbon atoms include N, P, O, S, and Si. Examples of heterocycloalkyl groups include groups derived from epoxide, azirine, thiirane, imidazolidine, morpholine, piperazine, piperidine, pyrazolidine, pyrrolidine, quinuclidine, and the like. In certain embodiments, heterocycloalkyl is _C5 heterocycloalkyl and is selected from pyrrolidinyl, tetrahydrofuranyl, tetrahydrothiophenyl, imidazolidinyl, oxazolidinyl, thiazolidinyl, doxolanyl, and dithiolanyl. In certain embodiments, heterocycloalkyl is _C6 heterocycloalkyl and is selected from piperidinyl, tetrahydropyranyl, piperidinyl, oxazinyl, dithianyl, and dioxanyl. In certain embodiments, the heterocycloalkyl group is C _3-6 heterocycloalkyl, C _3-5 heterocycloalkyl, C _5-6 heterocycloalkyl, and in certain embodiments C ₅ heterocyclo alkyl or _C6 heterocycloalkyl. In certain embodiments of heterocycloalkyl, the heteroatom groups are —O—, —S—, —NH—, —N(—CH ₃ )—, —SO—, and —SO ₂ —, certain In embodiments, the heteroatom group is selected from -O- and -NH-, and in certain embodiments the heteroatom group is -O- or -NH-.

An "agonist" binds to its complementary biologically active receptor or subunit(s) and elicits a biological response mediated by the receptor, or an existing receptor-mediated refers to a biologically active ligand that activates a receptor to enhance the biological activity of

"Affinity" refers to the strength of the binding interaction between a single biomolecule and its ligand/binding partner. Affinity is expressed as _IC50 .

"Antagonist" refers to a biologically active ligand or compound that binds to its complementary receptor or subunit(s) and blocks or reduces the biological response of the receptor.

Amino acid residues are abbreviated as follows: alanine is Ala or A, arginine is Arg is R, asparagine is Asn or N, aspartic acid is Asp or D, cysteine is Cys or C, glutamic acid is Glu or E, glutamine is Gln or Q, glycine is Gly or G, histidine is His or H, isoleucine is Ile or I, leucine is Leu or L. , lysine is Lys or K, methionine is Met or M, phenylalanine is Phe or F, proline is Pro or P, serine is Ser or S, threonine is Thr or T, Tryptophan is Trp or W, tyrosine is Tyr or Y, and valine is Val or V.

"Unnatural amino acids" include, for example, β-amino acids, homo-amino acids, proline and pyruvic acid derivatives, histidine derivatives with alkyl or heteroatom moieties attached to the imidazole ring, amino acids with pyridine-containing side chains, 3-substituted alanines derivatives, glycine derivatives, ring-substituted phenylalanine and tyrosine derivatives, and N-methyl amino acids.

Amino acids with large hydrophobic side chains include isoleucine (I), leucine (L), methionine (M), valine (V), phenylalanine (F), tyrosine (Y), and tryptophan (W).

Amino acids with small hydrophobic side chains include alanine (A), glycine (G), proline (P), serine (S), and threonine (T).

Amino acids with basic side chains include arginine (R), lysine (K), and histidine (H).

Amino acids with acidic side chains include aspartic acid (D) and glutamic acid (E).

Amino acids with polar/neutral side chains include histidine (H), asparagine (N), glutamine (Q), serine (S), threonine (T), and tyrosine (Y).

Amino acids with aromatic side chains include phenylalanine (F), histidine (H), tryptophan (W), and tyrosine (Y).

Amino acids with hydroxyl side chains include serine (S), threonine (T), and tyrosine (Y).

A "conservative amino acid substitution" means that each amino acid of the following groups may be replaced with another amino acid within the group: alanine (A), glycine (G), proline (P), serine ( S), and amino acids with small hydrophobic side chains, including threonine (T); amino acids with hydroxyl-containing side chains, including serine (S), threonine (T), and tyrosine (Y); aspartic acid (D) and Amino acids with acidic side chains including glutamic acid (E); polar neutral side chains including histidine (H), asparagine (N), glutamine (Q), serine (S), threonine (T), and tyrosine (Y) amino acids with basic side chains, including arginine (R), lysine (K), and histidine (H); isoleucine (I), leucine (L), methionine (M), valine (V), phenylalanine (F), amino acids with large hydrophobic side chains including tyrosine (Y) and tryptophan (W); and aromatic sides including phenylalanine (F), histidine (H), tryptophan (W), and tyrosine (Y). Amino acids with chains.

"PEG", "polyethylene glycol", and "poly(ethylene glycol)" refer to any non-peptidic, water-soluble poly(ethylene oxide). PEG can include the structure -(OCH ₂ CH ₂ ) _n -, where n is an integer from 1 to 4,000, for example. PEG can be -CH ₂ CH ₂ -O(CH 2 CH ₂ O) _n -CH ₂ CH ₂ - and/or _- (OCH Moieties such as ₂ CH ₂ ) _n O- moieties can also be included. PEG can be capped with suitable end groups. At least 50% of the repeating subunits of PEG can have the structure -CH ₂ CH ₂ -. PEGs can have any suitable molecular weight, structure, and/or geometry, such as branched, linear, forked, or multifunctional.

Molecular weight in the context of polymers refers to number average molecular weight as determined by gel permeation chromatography using polystyrene standards. The polymer has, for example, a polydispersity index (i.e., the number average molecular weight and weight average molecular weight of the polymer that are not equal to ).

Linker refers to a moiety that binds at least one IL-2R ligand, such as IL-2Rα ligand, IL-2Rβ ligand, and/or IL-2Rγc ligand. A linker can bind another IL-2R ligand, which can be the same IL-2R ligand or a different IL-2R ligand. A linker can also join one or more additional moieties that provide a desired physiological function. Linkers can be bivalent or multivalent. Linkers may be hydrolytically stable or may comprise physiologically hydrolyzable or enzymatically degradable linkages. Linkers can bind IL-2R ligands to form dimers, trimers, or higher order multiligand peptides (heteromers) and compounds.

A “physiologically cleavable” or “hydrolyzable” or “cleavable” bond is a bond that reacts with water (ie, is hydrolyzed) under physiological conditions. The tendency of a bond to hydrolyze in water will depend not only on the general type of bond connecting two central atoms, but also on the substituents attached to these central atoms. Suitable hydrolytically labile or weak linkages include, without limitation, carboxylate esters, phosphate esters, anhydrides, acetals, ketals, acyloxyalkyl ethers, imines, orthoesters, peptides, and oligonucleotides.

"Enzymatically degradable bond" means a bond that can be degraded or cleaved by one or more enzymes.

A "hydrolytically stable" bond or bond is substantially stable in water such that a chemical bond does not undergo hydrolysis to any significant degree over an extended period of time under physiological conditions, such as a covalent bond refers to the chemical bond of Examples of hydrolytically stable bonds include, but are not limited to: carbon-carbon bonds (eg, in aliphatic chains), ethers, amides, urethanes, and the like. Generally, a hydrolytically stable bond is a bond that exhibits a hydrolysis rate of less than about 1% to 2% per day under physiological conditions.

"IL-2Rα ligand" refers to a peptide capable of binding the IL-2Rα subunit of the mammalian IL-2 receptor with an affinity (IC ₅₀ ) of less than 100 µM.

"IL-2Rβ ligand" refers to a peptide capable of binding the IL-2Rβ subunit of the mammalian IL-2 receptor with an affinity (IC ₅₀ ) of less than 100 µM.

“IL-2Rγc ligand” refers to a peptide capable of binding the IL-2Rγc subunit of the mammalian IL-2 receptor with an affinity (IC ₅₀ ) of less than 100 μM.

"Human IL-2Rα ligand" is meant to refer to a peptide capable of binding the IL-2Rα subunit of the human IL-2 receptor with an affinity (IC ₅₀ ) of less than 100 μM.

A "human IL-2Rβ ligand" refers to a peptide capable of binding the IL-2Rβ subunit of the human IL-2 receptor with an affinity (IC ₅₀ ) of less than 100 μM.

A "human IL-2Rγc ligand" refers to a peptide capable of binding the IL-2Rγc subunit of the mammalian IL-2 receptor with an affinity (IC ₅₀ ) of less than 100 μM.

"Human IL-2Rα ligand" is meant to refer to a peptide capable of binding the IL-2Rα subunit of the human IL-2 receptor with an affinity (IC ₅₀ ) of less than 100 μM.

"IL-2R beta subunit" refers to the human (Homo sapiens) interleukin-2 receptor subunit beta precursor NCBI reference sequence NP_000689.1.

"IL-2Rγc subunit" refers to the human (Homo sapiens) interleukin-2 receptor subunit gamma precursor NCBI reference sequence NP_000197.1.

An "IL-2R ligand fusion protein" is one in which the translational reading frame of a ligand for a mammalian IL-2 receptor is fused to the open reading frame of another protein, i.e., an IL-2 receptor fusion partner, into a single set. Refers to a protein produced by recombinant DNA technology that produces a recombinant polypeptide. An IL-2R ligand fusion protein can include an IL-2Rβ ligand, an IL-2Rγc ligand, or both an IL-2Rβ ligand and an IL-2Rγc ligand. IL-2R ligand fusion proteins can be produced as disulfide-linked dimers linked together by disulfide bonds located in the hinge region. An IL-2R ligand fusion protein may comprise a peptide linker, eg, an amino acid sequence located between two proteins, including the fusion protein, such that the linker peptide sequence is not derived from either partner protein. can. Peptide linkers are incorporated into fusion proteins as spacers to promote proper protein folding and stability of the component protein moieties, improve protein expression, and/or enable better biological activity of the two fusion partners. can be done. Peptide linkers can be classified into the categories of unstructured flexible peptides or rigid structured peptides.

A bioisostere is an atom or molecule that fits the broadest definition of isostere. The concept of bioisosterism is based on the concept that single atoms, groups, moieties, or whole molecules having chemical and physical similarity produce similar biological effects. A bioisostere of a parent compound can still be recognized and accepted by its appropriate target, but its function is altered compared to the parent molecule. Parameters affected by bioisosteric substitution include, for example, size, conformation, induction and mesomeric effects, polarization properties, electrostatic interaction capacity, charge distribution, H-bond forming capacity, pKa (acidic), solubility , hydrophobicity, lipophilicity, hydrophilicity, polarity, potency, selectivity, reactivity, or chemical and metabolic stability, ADME (absorption, distribution, metabolism, and excretion). As is common in pharmaceuticals, a carboxyl or carboxylic acid functional group (--CO ₂ H) in a parent molecule is the desired one of the parent molecule having one or more carboxyl or carboxylic acid functional groups (--CO ₂ H). can be replaced by suitable substitutes or (bio)isosteres to overcome chemical or biological drawbacks while retaining the properties of

An "isostere" or "isosteric substitution" refers to any amino acid or other analogous moiety that has similar biochemical and/or structural properties as a particular amino acid. An "isostere" or "preferred isostere" of an amino acid is another amino acid of the same class, amino acids belonging to the following classes based on the propensity of their side chains to contact polar solvents, such as water: Hydrophobic (low tendency to contact water), polar or charged (energetically good contact with water). Examples of charged amino acid residues include lysine (+), arginine (+), aspartic acid (-), and glutamic acid (-). Examples of polar amino acids include serine, threonine, asparagine, glutamine, histidine, tyrosine. Exemplary hydrophobic amino acids include alanine, valine, leucine, isoleucine, proline, phenylalanine, tryptophan, cysteine, and methionine. The amino acid glycine has no side chain and is difficult to assign to one of the above classes. However, glycines are often found on the surface of proteins, often within loops, providing high flexibility to these regions, and isosteres may have similar characteristics. Prolines have the opposite effect, providing rigidity to the protein structure by adding specific torsion angles to segments of the polypeptide chain. An isostere can be a derivative of an amino acid, eg, a derivative having one or more modified side chains compared to a reference amino acid.

"Cyclization" means that one portion of a peptide or polypeptide molecule is linked to another portion of the peptide or polypeptide molecule, e.g., by forming a disulfide bridge or other similar bond, e.g., a lactam bond. Refers to a reaction that forms a closed ring. In certain embodiments, a monomeric subunit of a peptide monomeric compound or peptide dimeric compound described herein has a is cyclized through an intramolecular bond of

"Patient" refers to a mammal, eg, a human.

"Peptide" refers to a polymer whose monomers are α-amino acids linked together through amide bonds. A peptide can contain, for example, less than 100 amino acids, less than 50 amino acids, less than 40 amino acids, less than 30 amino acids, or less than 20 amino acids. Peptides can include naturally occurring alpha amino acids, non-naturally occurring amino acids, or combinations thereof.

In addition to peptides consisting solely of naturally occurring amino acids, peptidomimetics or peptide analogues are also provided. Peptide mimetics that are structurally similar to therapeutically useful peptides may be used to provide an equivalent or enhanced therapeutic or prophylactic effect. Generally, peptidomimetics have biological or pharmacological activity, such as paradigm peptides, eg, naturally occurring receptor binding peptides, by methods known in the art, but optionally —CH ₂ — NH—, —CH ₂ —S—, —CH ₂ —CH ₂ —, —CH═CH— (cis and trans), —COCH ₂ —, —CH(OH)CH ₂ —, and —CH ₂ SO—, etc. Structurally similar to peptides with one or more peptide bonds replaced by

Systematic substitution of one or more amino acids of a consensus sequence with a D-amino acid of the same type, such as D-lysine in place of L-lysine, can be used to generate more stable peptides. In addition, constrained peptides containing a consensus sequence or substantially identical consensus sequence mutations are capable of forming intramolecular disulfide bridges that cyclize the peptide by methods known in the art, e.g., internal cysteine residues can be generated by adding

A synthetic or non-naturally occurring amino acid refers to an amino acid that is not naturally occurring in vivo, yet can be incorporated into the peptide ligands provided by this disclosure. Suitable examples of synthetic amino acids include D-α-amino acids of naturally occurring L-α-amino acids, and non-naturally occurring D- and L-α-amino acids of the formula H ₂ NCHR ⁵ COOH. wherein R ⁵ is 1 on an aromatic nucleus selected from the group consisting of _C1-6 alkyl, C _3-8 cycloalkyl, C _3-8 heterocycloalkyl; hydroxyl, lower alkoxy, amino and carboxyl; Aromatic residue of 6 to 10 carbon atoms optionally with ~3 substituents; -alkylene-Y, where alkylene is an alkylene group of 1 to 7 carbon atoms, Y is hydroxyl, 1 to 3 substituents on the aromatic nucleus selected from the group consisting of hydroxyl, lower alkoxy, amino and carboxyl; aryl of 6 to 10 carbon atoms, such as; heterocyclic of 3 to 7 carbon atoms and 1 to 2 heteroatoms selected from the group consisting of oxygen, sulfur, and nitrogen; -C(O) R ² , wherein R ² is selected from the group consisting of hydrogen, hydroxy, lower alkyl, lower alkoxy, and —NR ³ R ⁴ , wherein R ³ and R ⁴ are independently hydrogen and lower alkyl; —S(O) _n R ⁶ , wherein n is 1 or 2 and R ² is C _1-6 alkyl, provided that R ⁵ is a naturally occurring amino acid undefined side chains).

Examples of other synthetic amino acids include amino acids, where the amino group is separated from the carboxyl group by more than one carbon atom, such as b-alanine, g-aminobutyric acid, and the like.

Examples of suitable synthetic amino acids include D-amino acids of naturally occurring L-amino acids, L-1-naphthyl-alanine, L-2-naphthylalanine, L-cyclohexylalanine, L-2-aminoisobutyric acid, methionine sulfoxide and sulfone derivatives, namely HOOC-(H ₂ NCH)CH ₂ CH ₂ -S(O) _n R ⁶ , where n and R ⁶ are as defined above, and lower alkoxy of methionine; Derivatives, namely HOOC-(H ₂ NCH)CH ₂ CH ₂ OR ⁶ where R ⁶ is as defined above.

"Polypeptide" refers to a polymer in which the monomers are alpha-amino acids linked together through amide bonds and containing more than 100 amino acids.

"N-terminus" refers to the end of a peptide or polypeptide, such as the N-terminus of an IL-2Rβ or IL-2Rγc ligand, having an amino group, as opposed to the carboxyl terminus having a carboxylic acid group.

"C-terminus" refers to the terminus of a peptide or polypeptide, such as the C-terminus of an IL-2Rβ or IL-2Rγc ligand, which has a carboxylic acid group, as opposed to the amino terminus, which has an amino group.

"Pharmaceutically acceptable" means approved or approved by federal or state governmental regulatory agencies, or the United States Pharmacopoeia or other generally recognized, for use in animals, and more particularly in humans. It refers to those listed in the pharmacopoeia.

"Pharmaceutically acceptable salt" refers to a salt of a compound that possesses the desired pharmacological activity of the parent compound. Such salts include inorganic acids as well as acid addition salts formed with one or more protonatable functional groups such as primary, secondary, or tertiary amines within the parent compound. Examples of inorganic acids include hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like. In certain embodiments, the salts are organic acids such as acetic acid, propionic acid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid. , tartaric acid, citric acid, benzoic acid, 3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethanedisulfonic acid, 2-hydroxyethanesulfonic acid, benzene sulfonic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, 4-toluenesulfonic acid, camphorsulfonic acid, 4-methylbicyclo[2.2.2]-oct-2-ene-1-carboxylic acid, glucoheptonic acid , 3-phenylpropionic acid, trimethylacetic acid, tertiary butyric acid, laurylsulfonic acid, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid, muconic acid and the like. In certain embodiments, when one or more acidic protons present in the parent compound are replaced by metal ions, such as alkali metal ions, alkaline earth ions or aluminum ions, or combinations thereof, or ethanolamine, Salts are formed when coordinated with organic bases such as diethanolamine, triethanolamine, N-methylglucamine. In certain embodiments, a pharmaceutically acceptable salt is the hydrochloride salt. In certain embodiments, a pharmaceutically acceptable salt is the sodium salt. In certain embodiments where a compound has more than one ionizable group, pharmaceutically acceptable salts include one or more counterions such as acid salts, eg, dihydrochloride salts.

The term "pharmaceutically acceptable salt" includes hydrates and other solvates as well as salts in crystalline or non-crystalline form. Where specific pharmaceutically acceptable salts are disclosed, the specific salt (e.g., hydrochloride) is an example of a salt and other salts may be formed using techniques known to those skilled in the art. Please understand. Additionally, one skilled in the art would be able to convert pharmaceutically acceptable salts to the corresponding compounds, free bases, and/or free acids using techniques generally known in the art. Also, Stahl and Wermuth, C.; G. (Editors), Handbook of Pharmaceutical Salts, Wiley-VCH, Weinheim, Germany, 2008.

"Pharmaceutically acceptable vehicle" means a pharmaceutically acceptable diluent, pharmaceutically acceptable adjuvant, pharmaceutically acceptable excipient, pharmaceutically acceptable carrier, or with which a compound provided by this disclosure can be administered to a patient at a dose sufficient to provide a therapeutically effective amount of the compound without destroying its pharmacological activity Non-toxic when administered.

A "solvate" refers to a molecular complex of a compound with stoichiometric or non-stoichiometric amounts of one or more solvent molecules. Such solvent molecules are those commonly used in the pharmaceutical arts and known to be harmless to patients, such as water, ethanol, and the like. A molecular complex of a compound or portion of a compound and a solvent can be stabilized by, for example, non-covalent intramolecular forces such as electrostatic forces, van der Waals forces, or hydrogen bonding. The term "hydrate" refers to solvates in which one or more of the solvent molecules is water.

"Pharmaceutical composition" means an IL-2R binding compound provided by the present disclosure or a pharmaceutically acceptable salt thereof, and at least one pharmaceutical composition administered with the compound or a pharmaceutically acceptable salt thereof to a patient. refers to a legally acceptable vehicle. Pharmaceutically acceptable vehicles are known in the art.

"Disease" refers to a disease, disorder, condition or symptom of any of the foregoing.

"Preventing" or "prevention" refers to reducing the risk of acquiring a disease or disorder (i.e., at least one of the clinical manifestations of a disease may expose or predispose to the disease, but not develop in patients who do not yet experience or display symptoms of disease). In some embodiments, "preventing" or "prevention" refers to reducing symptoms of a disease by taking a compound in a prophylactic manner. The application of therapeutic agents to prevent or prevent disease of a disorder is known as "prophylaxis."

"Prodrug" refers to a derivative of a therapeutic compound that requires a transformation within the body to release the active therapeutic compound. Prodrugs are often, but not necessarily, pharmacologically inactive until converted into the parent drug.

"Promoiety" refers to a group attached to a therapeutic compound, typically a functional group of the therapeutic compound, via a bond(s) that is cleavable under specified conditions of use. The bond(s) between drug and pro-moiety may be cleaved by enzymatic or non-enzymatic means. Under conditions of use, eg, after administration to a patient, the bond(s) between the therapeutic compound and the promoiety may be cleaved, releasing the parent therapeutic compound. Cleavage of the pro-moiety may proceed spontaneously, for example via a hydrolytic reaction, or it may proceed by another agent, such as by an enzyme, by light, by acid, or by physical or environmental parameters such as temperature. , pH, etc., or may be catalyzed or induced by exposure thereto. The drug may be endogenous to the conditions of use, eg, enzymes present in the systemic circulation of the patient to whom the prodrug is administered or the acidic conditions of the stomach, or the drug may be supplied exogenously.

"Substantially" means, for example, greater than 90%, greater than 95%, greater than 98%, or greater than 99%.

A "therapeutically effective amount" refers to an amount of a compound sufficient to treat a disease or a symptom thereof when administered to a subject to treat the disease or at least one of the clinical symptoms of the disease. . A "therapeutically effective amount" includes, for example, a compound, a disease and/or disease symptoms, the severity of the disease and/or the symptoms of the disease or disorder, the age, weight, and/or health of the patient being treated, and the prescribing physician's It can change depending on your judgment. The appropriate amount in any given case can be ascertained by those skilled in the art or determined by routine experimentation.

A "therapeutically effective dose" refers to a dose that provides effective treatment of a patient's disease or disorder. A therapeutically effective dose may vary from compound to compound and from patient to patient, depending on factors such as the patient's condition and route of delivery. A therapeutically effective dose can be determined according to routine pharmacological procedures known to those skilled in the art.

"Treating" or "treatment" of a disease means stopping or alleviating at least one of the disease or clinical symptoms of the disease or disorder, reducing the risk of acquiring the disease or at least one of the clinical symptoms of the disease , refers to reducing the onset of at least one disease or clinical symptom of a disease, or reducing or reducing the risk of developing at least one disease or clinical symptom of a disease. "Treating" or "treatment" can also mean treating a disease either physically (e.g., stabilization of identifiable symptoms), physiologically (e.g., stabilization of a physical parameter), or both. Refers to inhibiting as well as inhibiting at least one physical parameter or sign, which may or may not be discernible to the patient. In certain embodiments, "treating" or "treatment" means that the patient may be exposed to or be exposed to a disease or disorder even though the patient has not yet experienced or exhibited symptoms of the disease. Refers to delaying the onset of the disease or at least one or more symptoms thereof in a susceptible patient.

"Treg" or "Treg cell" refers to regulatory T cells. Regulatory T cells are a class of T cells that suppress the activity of other immune cells and are characterized by cell marker phenotypes CD4+/CD25+/FOXP3+, CD4+CD25+CD127lo, or CD4+/CD25+/FOXP3+/CD127lo using flow cytometry. Defined. Since FOXP3 is an intracellular protein and requires cell fixation and permeabilization for staining, the cell surface phenotype CD4+CD25+CD127lo- can be used to define viable Tregs. Tregs also include various Treg subclasses such as tTreg (thymus-derived) and pTreg (peripherally-derived, differentiated from naive T cells in the periphery). All Tregs express IL-2Rαβγ receptors, do not produce IL-2, and are dependent on IL-2 for proliferation. Those skilled in the art will recognize that Tregs are more potently activated by IL-2Rαβγc-biased agonists.

"CD4+ T cells" are lymphocytes that function to coordinate the immune response and fight infection by stimulating other immune cells such as macrophages, B lymphocytes (B cells), CD8 lymphocytes (CD8 cells). Kind. CD4+ T cells recognize peptides presented on MHC class II molecules found on antigen presenting cells.

"CD8+ (cytotoxic) T cells" are generated within the thymus and express T cell receptors. Cytotoxic T cells typically express a dimeric co-receptor CD8 comprising one CD8α and one CD8β chain. CD8+ T cells recognize peptides presented by MHC class 1 molecules found on all nucleated cells. CD8 heterodimers bind a conserved portion of MHC class 1 during T cell/antigen presenting cell interactions. CD8+ T cells (cytotoxic T lymphocytes, or CTLs) are important in immune defense against intracellular pathogens, including viruses and bacteria, and in tumor surveillance.

"NK (natural killer) cells" are lymphocytes within the same family as T-cells and B-cells and, as cells of the innate immune system, are classified as Group I innate lymphocytes (ILCs). NK cells respond to diverse pathological challenges, including killing virus-infected cells and detecting and controlling early signs of cancer.

"T cell functional activation" is defined as an IL-2 mediated response in T cells. Assays for functional activation of T cells include stimulation of pSTAT5, Treg cell proliferation or proliferation markers (such as Ki67), changes in immune cell type ratios, and stimulation of levels of T cell effector proteins.

Reference will now be made in detail to specific embodiments of compounds, compositions, and methods. The disclosed embodiments are not intended to limit the scope of the claims. On the contrary, the claims are intended to cover all alternatives, modifications and equivalents.

Interleukin-2 (IL-2) plays an important role in regulating immune responses and maintaining peripheral self-tolerance by having both immunostimulatory and immunomodulatory functions. IL-2 acts primarily as a T cell growth factor essential for T cell proliferation and survival and the generation of effector and memory T cells. IL-2 is a four alpha-helical bundle cytokine that belongs to a family of structurally related cytokines that includes IL-4, IL-7, IL-9, IL-15, and IL-21. IL-2 is produced by activated CD4+ T cells in response to antigenic stimulation and can be produced by CD8+ T cells and innate immune cells such as activated dendritic cells (DC) and natural killer (NK) cells.

IL-2 binds to various forms of the IL-2 receptor (IL-2R), particularly the monomeric, dimeric and trimeric forms. Monomeric IL-2R consists of the membrane-associated IL-2Rα (CD25) chain, which also exists in a soluble form but is incapable of inducing signaling events. The trimeric IL-2R consists of IL-2Rα, IL2Rβ (CD122), and IL-2Rγc, also known as common γ chain (γc) or CD132, shared by all members of the IL-2 cytokine family. be. Dimeric IL-2R comprises IL-2Rγc and IL-2Rβ subunits. In contrast to the monomeric IL-2R, both dimeric and trimeric IL-2 receptors effect downstream signaling cascades upon IL-2 binding. IL-2 binds with high affinity to trimeric IL-2R, but with low to moderate affinity to dimeric IL-2R, reducing the sensitivity of cells to IL-2. change. In addition, IL-2 is expressed on the surface of activated dendritic cells for transpresentation to neighboring cells, including antigen-specific naive T cells and NK cells, which express both IL-2Rβ and IL-2Rγc subunits. It is capable of binding to IL-2Rα expressed above. This transpresentation of IL-2 has been shown to promote early high-affinity IL-2 signaling that is required early in the immune response to prime naive T cells to produce IL-2. .

IL-2 is first captured by IL2Rα, causing a conformational change in IL-2 and increasing its affinity for IL-2Rβ. association of IL-2 with IL-2Rβγc induces dimerization of signaling motifs in the cytoplasmic tails of IL-2Rβ and IL-2Rγc, resulting in phosphorylation/activation of Janus kinases, JAK1 and JAK3; It then exerts kinase activity on key tyrosine residues in the tail of the IL-2Rβ subunit.

Downstream signaling occurs through three major pathways, the JAK-STAT pathway, the phosphoinositide 3-kinase (PI3K)-AKT pathway, and the mitogen-activated protein kinase (MAPK) pathway. These pathways ultimately, through recruitment of the adapter protein Shc and the transcription factor STAT5, lead to transcription of target genes that contribute to IL-2-dependent biological effects. Target genes for IL-2 signaling include cyclin D2, bcl-2, fasL, cd25 (encoding IL-2Rα), socs1-2, and the IL-2 silencing gene prdm1, which encodes the transcription factor BLIMP1. mentioned. Production of the IL-2 negative regulator BLIMP1 is essential for maintaining the balance between effector T cells and Treg cells that are important for immune homeostasis.

IL-2 plays a dual role in T cell activation by stimulating T cell proliferation and differentiation and by maintaining and expanding the population of immunosuppressive Treg cells. Conventional naive CD4+ and CD8+ T cells express dimeric IL-2R and thus require high concentrations of IL-2 to induce their initial proliferation. Upon activation, these T cells express the high-affinity trimeric IL-2R, promoting differentiation of T cells into either effector (Teff) or memory cells. This differentiation depends on the strength and duration of the IL-2 signal.

During primary expansion of CD8+ T cells in the presence of low to moderate levels of IL-2, a subset of CD8+ T cells differentiate into memory T cells. Cells do this by downregulating CD25 and upregulating CD127 (IL-7R) and CD62 (L-selectin), a key receptor for secondary responses upon reinfection. During acute infection, sustained high levels of IL-2 lead to rapid upregulation of CD25 and differentiation of CD8+ cells into cytotoxic effector cells. Upregulation induces IL-2-driven expression of the death receptors fas and fasL, causing activation-induced cell death (AICD) upon pathogen clearance. For CD4+ T cells, activation of STAT5 signaling by IL-2 leads to their differentiation into multiple helper T cell populations, including Th1, Th2, and Th17, by modulating the expression of appropriate receptors for each response. affect.

Constitutive or background levels of IL-2 are essential for Treg cell survival and function by maintaining expression of FOXP3 and CD25. Treg cells naturally develop within the thymus and are activated upon contact with self-peptides. In addition, Treg cells can be generated by conventional stimulation of CD4+ T cells in interacting with antigens in peripheral lymphoid organs. Treg cells do not produce IL-2 and therefore rely on IL-2 producing cells such as conventional T cells. In addition, due to their high expression of IL-2Rα (CD25), Tregs are able to consume and limit systemic levels of IL-2, ensuring regulation of immune balance. In the absence of IL-2, Treg cell numbers decrease and effector T cell numbers increase, resulting in increased susceptibility to autoimmune and inflammatory disorders. Thus, the unique activation of Treg cells with low levels of IL-2 that did not activate CD4+ or CD8+ T cells allowed the development of IL-2 as a potential therapeutic agent in autoimmune and inflammatory diseases.

The production of IL-2 from both arms of the immune system underscores the importance of this cytokine early in infection, as well as in secondary adaptive responses. Furthermore, due to IL-2's dual functions in both protective immunity and immune tolerance, IL-2 appears to be both an immunostimulatory and an immunosuppressive agent for cancer and autoimmune diseases, respectively. It could be a potential therapeutic agent in various therapies.

The present disclosure is directed to IL-2Rβ ligands, IL-2Rγc ligands, and compounds comprising IL-2Rβ ligands and/or IL-2Rγc ligands. Compounds containing IL-2Rβ ligands and IL-2Rγc ligands can be IL-2R agonists, including selective IL-2R agonists.

The IL-2Rβ and IL-2Rγc ligands provided by the present disclosure simultaneously mask peripheral biological activity, target delivery to tumors, selectively activate cytotoxic anti-tumor cells, and promote cytotoxicity at tumor sites. It contains peptide domains suitable for strategies to induce IL-2 receptor activation. IL-2Rβ and IL-2Rγc ligands provided by the present disclosure, and compounds comprising IL-2Rβ and/or IL-2Rγc ligands, can also be used to treat autoimmune diseases.

IL-2Rα ligands and compounds containing IL-2Rα ligands are disclosed in US Provisional Application No. 62/856,305, filed June 3, 2019, which is incorporated by reference in its entirety.

Compounds containing IL-2R agonists and IL-2Rβ and/or IL-2Rγc ligands can be designed to selectively activate particular forms of the IL-2 receptor. Small peptide IL-2R ligands with amino acid sequences unrelated to those of natural cytokines selectively bind and activate IL-2Rβ and/or IL-2Rγc subunits, resulting in therapeutic IL-2 activity. However, the limited clinical success of IL-2-based compounds in cancer therapy can be circumvented. Because IL-2R peptide ligands are small, ie, 5-30 amino acids, and have very low immunogenic potential, small peptide IL-2R ligands can be incorporated into compounds to enhance therapeutic efficacy. For example, this allows the affinity of IL-2R peptide ligands for each of the three IL-2R subunits to be tailored and thereby chemically targeted to induce responsiveness of specific immune cell populations. Provides flexibility to the tumor site.

Peptide ligands for the IL-2Rβ and IL-2Rγc subunits can be identified from highly complex peptide diversity libraries, such as phage display libraries, optimized by peptide synthesis and can be monomeric, homo-oligomeric, heterozygous. It can be assembled into dimers, or other compounds, and the IL-2Rβ and IL-2Rγc binding subunits can be designed to be competent signaling structures.

Small agonists of IL-2R provided by the present disclosure can activate cells that do not express IL-2Rα subunits with similar potency as cells that do express IL-2Rα subunits, thereby activating cells such as Tregs. Avoid preferential activation of cells expressing IL-2Rα. These IL-2R agonists are termed selective IL-2Rβγc agonists. Since the selective IL-2R agonists provided by the present disclosure can be chemically synthesized, the IL-2R agonists can be administered to each receptor in order to optimize potency and efficacy and improve metabolic stability. Modifications can be made using natural and/or unnatural amino acids to independently adjust the binding affinity for the subunits. Small agonists also allow such modifications to be made with less potential to induce immunogenicity. Also, because of their chemical plasticity, peptides can be “caged” to construct reversibly inactive prodrugs using complexes that are sensitive to tumor-specific environmental triggers, such as proteases, or low pH. can be. For example, the pH-dependent binding properties of peptides can be optimized by using unnatural amino acids with side chain ionizable groups with pKa's in the range of pH 5.0 to pH 8.0. As with proteins, pharmacokinetic-enhancing moieties such as PEG can be added to peptides either as part of a "cage" strategy or independently of a "cage" strategy. Finally, sites on the peptide can be reserved for attachment of various tumor-targeting moieties such as tumor-specific antibodies. These characteristics of IL-2Rβγc agonists can be exploited in designing optimal therapeutic candidates based on IL-2Rβγc agonists provided by this disclosure.

Certain compounds provided by this disclosure include ligands that selectively activate certain forms of the IL-2 receptor. These agonists can stimulate cytotoxic tumor target cell populations without inducing Treg immunosuppressive activity at the tumor site. Cellular selectivity of agonists can enhance the effectiveness of IL-2 for anti-tumor therapy.

Peptides with binding affinity for the IL-2Rβγc subunit can be identified by random peptide diversity generation systems, eg, using peptides on plasmids or peptides on phage systems, in conjunction with the affinity enrichment process. Synthetic peptide library technology, such as DNA-encoded peptide libraries, can also be used.

Using such systems, random peptides are generally designed to have a defined number of amino acid residues in length, such as 6-20 amino acids. To generate a collection of oligonucleotides encoding random peptides, codon motifs (NNK) _x , where N is the nucleotide A, C, G, or T (equimolar; depending on the method used) , other nucleotides can be used), K is G or T (equimolar), and x is an integer corresponding to the number of amino acids in the peptide, eg, 6-20), the NNX motif ( 1 of every 12 amino acids, 2 of every 5 amino acids, 3 of every 3 amino acids, and only one of the three stop codons). can be used for Thus, the NNK motif encodes all amino acids and only one stop codon, reducing codon bias.

In the systems employed, random peptides are placed on the surface of phage particles, as part of fusion proteins containing either the pIII or pVIII coat proteins of phage fd derivatives (peptides on phage), or as LacI peptides bound to plasmids. It can be presented as a fusion protein with a fusion protein (peptide on a plasmid). Phage or plasmids containing DNA encoding peptides can be identified and isolated by an affinity enrichment process using immobilized IL-2Rβ and/or IL-2Rγc subunits. The affinity enrichment process, sometimes referred to as "panning," involves incubating phage or plasmids with immobilized receptor, collecting phage or plasmids (along with their associated DNA) that bind to the receptor, and harvesting Include multiple rounds to produce more phage or plasmid (with accompanying LacI-peptide fusion protein). The extracellular domain (ECD) of the IL-2Rβ and/or IL-2Rγc subunits can be used during panning. Engineered forms of IL-2Rβ and IL-2Rγc can be expressed in host cells such as CHO cells. After receptor collection, receptors can be tested for binding to IL-2Rβ or IL-2Rγc specific phage clones. Peptides can also be identified by panning for IL-2R subunits fused to the Fc domain. IL-2 receptor, and its extracellular domain, can be produced in recombinant host cells.

After several rounds of affinity enrichment, the phage or plasmid and associated peptides can be tested by ELISA to determine whether the peptide specifically binds to IL-2Rβ and/or IL-2Rγc subunits. The assay is described for an affinity enrichment process, except that unbound phage can be removed, followed by treatment with antibodies such as rabbit anti-phage antibodies, followed by alkaline phosphatase (AP)-conjugated goat anti-rabbit antibodies. It can be done using methods similar to those in The amount of alkaline phosphatase in each well can be determined by standard methods.

By comparing test wells to control wells without IL-2 receptor, it can be determined whether the fusion protein specifically binds to the receptor. Phage pools found to bind IL-2Rβ and/or IL-2Rγc subunits can be screened in a colony lift probe format using radiolabeled monovalent receptors. This probe can be generated using protein kinase A to phosphorylate a peptide sequence fused to the C-terminus of the soluble receptor. The receptor can then be labeled with ³³ P to high specific activity for use as a monovalent probe to identify high affinity ligands using colony lift.

Peptides found to specifically bind to the IL-2Rβ subunit or IL-2Rγc subunit can then be synthesized as free peptides (eg, without phage) and tested in blocking assays. Blocking assays are performed in a similar manner to ELISAs, except that IL-2Rβ or IL-2Rγc binding peptides or reference peptides can be added to wells prior to tracer compounds that can detect receptor binding. can do. Examples of tracer compounds include specific phage clones with receptor binding peptides, radiolabeled or biotinylated peptides known to bind receptors, or labeled variants of IL-2. Peptides that block tracer binding to IL-2Rβ or IL-2Rγc are preferred compounds of the invention.

When using random peptide generation systems that allow multivalent ligand-receptor interactions, the density of immobilized receptors is important in determining the affinity of ligands that can bind to immobilized receptors. can be a factor. At higher receptor densities, e.g., when each anti-receptor antibody-coated well is treated with 0.25 mg to 0.5 mg of receptor, lower receptor densities (e.g., each anti-receptor antibody-coated wells treated with 0.5-1 ng of receptor), multivalent binding is more likely to occur. If multivalent binding is occurring, the relatively low A ligand with affinity will be more likely to be isolated.

A screening method that can be used to identify peptides that bind IL-2R is to first identify a lead peptide that binds to the extracellular domain of the receptor and then synthesize other peptides that are similar to the lead peptide. can include doing For example, the peptide on phage system can be used to screen random libraries to discover phage displaying peptides that bind to IL-2Rβ and/or IL-2Rγc subunits. Phage DNA can be sequenced to determine the sequence of peptides displayed on the surface of the phage.

For example, clones capable of specifically binding IL-2R can be identified from random linear libraries. The sequences of these peptides can serve as the basis for the construction of other peptide libraries designed to frequently contain derivatives of the peptides originally identified. These libraries can be synthesized to favor the production of peptides that differ at only a few residues from the binding peptide. This approach uses a mixture of the four nucleoside triphosphates (i.e., 55% of the "correct" nucleotides, and the other three 15% of each of the nucleotides is one preferred mixture for this purpose, 70% of the "correct" nucleotides and 10% of each of the other 3 nucleotides is another preferred mixture for this purpose) is used to synthesize oligonucleotides containing the connecting peptide-encoding sequences, except that derivatives of the connecting peptide-encoding sequences can be generated.

Generate a "thematic mutagenesis" library based on the consensus sequence and derivatize the lead peptide by mutating with 5 NNK codons at each end at a frequency of 70:10:10:10 Various strategies can be used for this (probe with radiolabeled monovalent receptor, and with or without peptide elution).

The "peptide on plasmid" method can also be used for peptide screening and mutagenesis studies. According to this approach, random peptides can be fused at the C-terminus of LacI through expression from a plasmid vector carrying the fusion gene. Binding of the LacI-peptide fusion to its encoding DNA occurs via the lacO sequence on the plasmid, resulting in stable peptide-LacI-plasmid complexes that can be screened by affinity purification (panning) on immobilized receptors. form the body. The plasmid thus isolated is then cloned into E. coli by electroporation. E. coli to amplify selected populations for additional rounds of screening or for examination of individual clones.

Additionally, random peptide screening and mutagenesis studies can be performed using a modified C-terminal Lac-I display system that can reduce display value. Libraries can be screened and the resulting DNA inserts can be cloned as pools into maltose binding protein (MBP) vectors, allowing their expression as C-terminal fusion proteins. Crude cell lysates from randomly picked individual MBP fusion clones can then be assayed for IL-2R binding in an ELISA format.

Various methods can be used to assess _IC50 binding affinity values. For example, using a competitive binding ELISA assay, an IL-2 tracer, an IL-2Rβ ligand tracer, or an IL-2Rγc ligand tracer, the peptide inhibits binding of the tracer to the extracellular domain of the IL-2 receptor. can determine whether _IC50 values can be determined using free peptides, which can optionally be C-terminally amidated or prepared as esters or other carboxamides. One or two glycine residues may precede the N-terminal and C-terminal amino acids of the synthetic peptide to reproduce the exact sequence displayed by the phage. These glycines are not believed to be required for binding or activity.

Generally, peptides and peptidomimetics with IC ₅₀ greater than about 100 μM lack sufficient binding affinity (IC ₅₀ ) to be useful for imaging, targeting, diagnostic, and therapeutic applications. For imaging, targeting, and diagnostic purposes, peptides and peptidomimetics can have an _IC50 of, for example, about 1 μM or less; can have an _IC50 of less than, or less than 100 nM.

A robust and quantitative bioassay can be used to characterize the biological activity of compounds exhibiting IL-2Rβγc agonist activity. The assay measures activation of the IL-2 receptor, measures downstream functional consequences of IL-2R activation, and/or measures therapeutically relevant consequences and functions of activated IL-2 receptor. can do. This assay can be used to measure the therapeutic activity and efficacy of IL-2R selective agonists and compounds comprising IL-2Rβ and/or IL-2Rγc ligands, and IL-2 selective agonists in animals or humans. can also be used to measure the pharmacodynamics of

Activation of the IL-2Rβγc subunit can be determined using a suitable assay. One example is an assay that can measure phosphorylation of the signaling protein STAT5, flow cytometry as measured by an antibody specific for the phosphorylated protein (pSTAT5). Phosphorylation of STAT5 is an essential step in the IL-2 signaling pathway. Therefore, measurement of phosphorylated STAT5 (pSTAT5) in cells is recognized as reflecting IL-2 activation in these cells, and given appropriate exposure times and conditions, other effects of IL-2 treatment would predict biological consequences.

Another assay for assessing functional activation is to use an assay that measures IL-2 stimulated proliferation of cells. Cell proliferation is measured, for example, by measuring tritiated thymidine incorporation into a purified T cell population, using flow cytometry to measure an increase in the number of cells of a particular T cell type in a mixed population of cells. or by measuring the cell division-related dilution of an adult fluorochrome such as carboxyfluorescein succinimidyl ester (CFSE) by flow cytometry.

IL-2Rβ ligands provided by the present disclosure can exhibit binding affinities (IC ₅₀ ) for human IL-2Rβ subunits of less than 100 μM, less than 10 μM, less than 1 μM, less than 0.1 μM, or less than 0.01 μM. can.

IL-2Rβ ligands can exhibit binding affinities (IC ₅₀ ) for human IL-2Rβ subunits of 1 pM to 100 μM, 10 pM to 10 μM, 100 pM to 1 μM, 0.001 μM to 1 μM, or 0.01 μM to 1 μM. can.

IL-2Rβ ligands provided by the present disclosure exhibit binding affinities (IC ₅₀ ) for mammalian IL-2Rβ subunits of less than 100 μM, less than 10 μM, less than 1 μM, less than 0.1 μM, or less than 0.01 μM be able to.

The IL-2Rβ ligand should exhibit a binding affinity (IC ₅₀ ) for the mammalian IL-2Rβ subunit of 1 pM to 100 μM, 10 pM to 10 μM, 100 pM to 1 μM, 0.001 μM to 1 μM, or 0.01 μM to 1 μM. can be done.

IL-2Rβ ligands provided by the present disclosure have less than 100 μM, less than 10 μM, less than 1 μM, less than 0.1 μM, or less than 0.01 μM binding to human IL-2Rβ subunit and human IL-2Rγc subunit, respectively. Affinity ( _IC50 ) can be shown.

IL-2Rβ ligands have binding affinities of 1 pM to 100 μM, 10 pM to 10 μM, 100 pM to 1 μM, 0.001 μM to 1 μM, or 0.01 μM to 1 μM for human IL-2Rβ subunit and human IL-2Rγc subunit, respectively. properties ( _IC50 ).

IL-2Rβ ligands can exhibit binding affinities (IC ₅₀ ) for human IL-2Rα (CD25) subunits greater than 100 μM, greater than 1 mM, greater than 10 mM, or greater than 100 mM.

The IL-2Rβ ligand has at least 10-fold, at least 50-fold, at least 100-fold, at least 500-fold, or at least 1,000-fold or more the binding affinity (IC ₅₀ ) of the IL-2Rβ ligand for the human IL-2Rα subunit. A certain binding affinity (IC ₅₀ ) for the human IL-2Rβ subunit can be demonstrated.

The IL-2Rβ ligand can comprise an amino acid sequence selected from SEQ ID NOS:1-193, SEQ ID NOS:578-903, and SEQ ID NOS:1028-1050.

The IL-2Rβ ligand can comprise an amino acid sequence selected from SEQ ID NOs:1-193, SEQ ID NOs:578-903, and SEQ ID NOs:1028-1050, independently with the following conservative substitutions: Amino acids with small hydrophobic side chains, including alanine (A), glycine (G), proline (P), serine (S), or threonine (T), or tyrosine (Y); serine (S), threonine (T ); amino acids with acidic side chains, including aspartic acid (D), or glutamic acid (E); histidine (H), asparagine (N), glutamine (Q), serine (S) , threonine (T), or tyrosine (Y); amino acids with basic side chains, including arginine (R), lysine (K), or histidine (H); isoleucine (I ), leucine (L), methionine (M), valine (V), phenylalanine (F), tyrosine (Y), or tryptophan (W); and phenylalanine (F), histidine. It may include one or more of amino acids with aromatic side chains, including (H), tryptophan (W), or tyrosine (Y).

The IL-2Rβ ligand can have the amino acid sequence of formula (1) (SEQ ID NO:1), the amino acid sequence of formula (1a) (SEQ ID NO:2), or the amino acid sequence of formula (1b) (SEQ ID NO:3);
^{-X3 -X4 -X5 -X6 -X7 -X8 -X9 -X10} - (1)
^-X2 -CX3 ^{-X4 -X5 -X6 -X7 -X8 -X9 -X10 -CX11} - ( ^1a )
^{-X1 -X2} -CX3 ^{-X4 -X5 -X6 -X7 -X8 -X9 -X10 -CX11 -X12 -} (1b)
wherein X ¹ is selected from A, D, E, F, G, I, K, L, M, N, P, Q, S, T, V, W, and ^Y ; , C, D, E, F, G, H, K, L, N, P, R, S, T, W, and Y, and X ³ are selected from A, D, E, F, G, H , M, N, Q, R, S, T, W, and Y, and X ⁴ is selected from A, D, E, F, G, I, K, L, M, N, Q, R, S , T, V, and Y, X ⁵ is selected from A, G, I, Q, S, T, V, and W, X ⁶ is selected from A, D, E, G, H, K , L, M, N, P, Q, R, S, T, and V; X ⁷ is selected from F, I, K, L, Q, and V; X ⁸ is selected from D, F , G, H, M, N, W, and Y, X ⁹ is selected from A, D, E, M, P, Q, S, T, V, and W, and X ¹⁰ is selected from D , F, I, L, M, S, T, V, and Y, and X ¹¹ is selected from D, E, F, H, I, L, M, Q, S, T, V, W, and Y and X ¹² is selected from F, I, L, M, N, S, V, W, and Y;

In the IL-2Rβ ligands of formulas (1)-(1b), ^X1 may be selected from F, I, L, M, and V.

In the IL-2Rβ ligands of formulas (1)-(1b), ^X2 may be selected from D, E, F, G, H, L, N, P, R, S, T, W, and Y.

In the IL-2Rβ ligands of formulas (1)-(1b), ^X5 can be A.

In the IL-2Rβ ligands of formulas (1)-(1b), ^X6 may be selected from D, E, and Q.

In the IL-2Rβ ligands of formulas (1)-(1b), ^X7 may be selected from F, I, L, and V.

In the IL-2Rβ ligands of formulas (1)-(1b), ^X8 can be G.

In the IL-2Rβ ligands of formulas (1)-(1b), ^X9 may be selected from D, E, and Q.

X ¹⁰ may be selected from F, I, L, M, V, and Y in the IL-2Rβ ligands of formulas (1)-(1b).

In IL-2Rβ ligands of formulas (1)-(1b), X ¹¹ may be selected from D and E.

In the IL-2Rβ ligands of formulas (1)-(1b), ^X12 may be selected from F, I, L, M, and V.

In the IL-2Rβ ligands of formulas (1)-(1b), X ¹ can be selected from F, I, L, M, and V, X ² can be D, E, F, G, H, L, N, P, R, S, T, W, and Y, and X ³ are A, D, E, F, G, H, M, N, Q, R, S, T , W, and Y, and X ⁴ is selected from A, D, E, F, G, I, K, L, M, N, Q, R, S, T, V, and Y X ⁵ can be A, X ⁶ can be selected from D, E, and Q, X ⁷ can be selected from F, I, L, and V , X ⁸ can be G, X ⁹ can be selected from D, E, and Q, and X ¹⁰ can be selected from F, I, L, M, V, and Y , X ¹¹ can be selected from D and E, and X ¹² can be selected from F, I, L, M, and V.

The IL-2Rβ ligand can comprise the amino acid sequence of formula (1) (SEQ ID NO:1), the amino acid sequence of formula (1a) (SEQ ID NO:2), or the amino acid sequence of formula (1b) (SEQ ID NO:3);
^{-X3 -X4 -X5 -X6 -X7 -X8 -X9 -X10} - (1)
^-X2 -CX3 ^{-X4 -X5 -X6 -X7 -X8 -X9 -X10 -CX11} - ( ^1a )
^{-X1 -X2} -CX3 ^{-X4 -X5 -X6 -X7 -X8 -X9 -X10 -CX11 -X12 -} (1b)
wherein X ¹ can be selected from amino acids, X ² can be selected from amino acids, X ³ can be selected from amino acids, X ⁴ can be selected from amino acids, X ⁵ can be selected from Can be selected from amino acids containing small hydrophobic side chains, X ⁶ can be selected from amino acids, X ⁷ can be selected from amino acids containing large hydrophobic side chains, X ⁸ can be selected from small hydrophobic can be selected from amino acids containing side chains, X ⁹ can be selected from amino acids containing polar neutral or acidic side chains, X ¹⁰ can be selected from amino acids containing large hydrophobic side chains, X ¹¹ can be selected from amino acids and X ¹² can be selected from amino acids containing large hydrophobic side chains.

In the IL-2Rβ ligands of formulas (1)-(1b), X ¹ can be selected from amino acids containing large hydrophobic side chains, X ² can be selected from amino acids, and X ³ can be selected from can be selected from amino acids, X ⁴ can be selected from amino acids, X ⁵ can be selected from amino acids containing small hydrophobic side chains, X ⁶ can be selected from polar neutral or acidic side chains X ⁷ can be selected from amino acids containing large hydrophobic side chains, X ⁸ can be selected from amino acids containing small hydrophobic side chains, X ⁹ can be selected from amino acids containing polar neutral or acidic side chains, X ¹⁰ can be selected from amino acids containing large hydrophobic side chains, and X ¹¹ can be selected from amino acids containing polar neutral or acidic side chains. and X ¹² can be selected from amino acids containing large hydrophobic side chains.

In the IL-2Rβ ligands of formulas (1)-(1b), X ¹ can be selected from I, L, M, V, F, W, and Y, and X ² can be selected from amino acids. X ³ can be selected from amino acids, X ⁴ can be selected from amino acids, X ⁵ can be selected from A, G, P, S, and T, and X ⁶ can be selected from , H, N, Q, S, T, Y, D, and E; X ⁷ may be selected from I, L, M, V, F, W, and Y; ⁸ can be selected from A, G, P, S, ^and T; X ⁹ can be selected from H, N, Q, S, T, Y, D, and E; , I, L, M, V, F, W, and Y; X ¹¹ may be selected from H, N, Q, S, T, Y, D, and E; ¹² can be selected from I, L, M, V, F, W, and Y;

In the IL-2Rβ ligands of formulas (1)-(1b), X ¹ can be selected from I, L, M, V, F, W, and Y, and X ² can be selected from amino acids. X ³ can be selected from amino acids, X ⁴ can be selected from amino acids, X ⁵ can be A, X ⁶ can be H, N, Q, S, T , Y, D, and E; X ⁷ can be selected from I, L, M, V, F, W, and Y; X ⁸ can be G; X ⁹ can be selected from H, N, Q, S, T, Y, D, and E; X ¹⁰ can be selected from I, L, M, V, F, W, and Y; X ¹¹ can be selected from H, N, Q, S, T, Y, D, and E, and X ¹² can be selected from I, L, M, V, F, W, and Y can do.

In the IL-2Rβ ligands of formulas (1)-(1b), ^X1 may be selected from I, L, M, and V.

In the IL-2Rβ ligands of formulas (1)-(1b), ^X2 may be selected from D and E.

In IL-2Rβ ligands of formulas (1)-(1b), ^X6 may be selected from Q, E, and D.

In the IL-2Rβ ligands of formulas (1)-(1b), ^X7 may be selected from V, L, and I.

In the IL-2Rβ ligands of formulas (1)-(1b), ^X9 may be selected from E, D, and Q.

X ¹⁰ may be selected from L, V, I, and Y in the IL-2Rβ ligands of formulas (1)-(1b).

In IL-2Rβ ligands of formulas (1)-(1b), X ¹¹ may be selected from D and E.

In the IL-2Rβ ligands of formulas (1)-(1b), ^X12 may be selected from L, I, and F.

In the IL-2Rβ ligands of formulas (1)-(1b), X ¹ can be selected from L, I, F, and V, X ² can be selected from D and E, X ⁶ can be selected from Q, E, and D, X ⁷ can be selected from V, L, and I, X ⁸ can be G, X ⁹ can be E, D , and Q, X ¹⁰ can be selected from L, V, I, and Y, X ¹¹ can be selected from D and E, X ¹² can be selected from L, I , and F.

In the IL-2Rβ ligands of formulas (1)-(1b), X ¹ can be selected from F, I, M, and Y, and X ² can be selected from E, D, and R. , X ³ can be selected from and amino acids, X ⁴ can be selected from amino acids, X ⁵ can be A, and X ⁶ can be selected from A, P, and Q X ⁷ can be selected from I and V, X ⁸ can be G, X ⁹ can be selected from E and Q, X ¹⁰ can be selected from I, L , and V, X ¹¹ can be selected from E, D, and Q, and X ¹² can be selected from I and L.

IL-2Rβ ligands may comprise an amino acid sequence selected from any one of SEQ ID NO:4-SEQ ID NO:163.

The IL-2Rβ ligand can comprise an amino acid sequence selected from any one of SEQ ID NOs: 1-163, wherein the amino acid sequence is on the N-terminus, the C-terminus, or both the N-terminus and the C-terminus. It may end with the above amino acids -GG-.

The IL-2Rβ ligand may comprise an amino acid sequence selected from any one of SEQ ID NOs: 1-163, each amino acid independently having the following conservative substitutions: alanine (A), glycine (G), amino acids with small hydrophobic side chains including proline (P), serine (S), or threonine (T); hydroxyl-containing sides including serine (S), threonine (T), or tyrosine (Y) amino acids with acidic side chains, including aspartic acid (D), or glutamic acid (E); histidine (H), asparagine (N), glutamine (Q), serine (S), threonine (T), or amino acids with polar neutral side chains, including tyrosine (Y); amino acids with basic side chains, including arginine (R), lysine (K), or histidine (H); isoleucine (I), leucine (L) , methionine (M), valine (V), phenylalanine (F), tyrosine (Y), or tryptophan (W); and phenylalanine (F), histidine (H), tryptophan ( W), or amino acids with aromatic side chains, including tyrosine (Y).

IL-2Rβ ligands of SEQ ID NOs: 1-163 exhibit affinities for IL-2Rβ subunits of less than 100 μM.

The IL-2Rβ ligand can comprise the amino acid sequence of formula (2) (SEQ ID NO: 164), the amino acid sequence of formula (2a) (SEQ ID NO: 165), or the amino acid sequence of formula (2b) (SEQ ID NO: 166);
^{-X15 -X16 -X17 -X18 -X19 -X20 -X21 -X22} - (2)
^{-X14 -CX15 -X16 -X17 -X18 -X19 -X20 -X21 -X22 -CX23} - (2a)
^{-X13 -X14 -CX15 -X16 -X17 -X18 -X19 -X20 -X21 -X22 -CX23 -X24} - (2b)
wherein X ¹³ can be selected from A, D, E, G, N, Q, R, and V; and X ¹⁴ can be selected from E, F, I, L, M, and Q; , X ¹⁵ can be selected from D, G, L, and N, X ¹⁶ can be selected from L, P, V, and Y, and X ¹⁷ can be selected from F, G, and M, X ¹⁸ can be selected from A, D, N, and Q; X ¹⁹ can be selected from F, I, L, S, V, W, and Y; X ²⁰ can be selected from D and W; X ²¹ can be selected from P and Y; X ²² can be selected from A, D, Q, and S; ²³ can be selected from I, L, Q, W, and Y; X ²⁴ can be selected from E, F, I, L, T, V, and W;

X ¹⁶ can be V in the IL-2Rβ ligands of formulas (2)-(2b).

X ¹⁷ can be G in the IL-2Rβ ligands of formulas (2)-(2b).

X ²⁰ can be W in the IL-2Rβ ligands of formulas (2)-(2b).

In the IL-2Rβ ligands of formulas (2)-(2b), ^X21 can be P.

In the IL-2Rβ ligands of formulas (2)-(2b), X ¹³ can be selected from E, N, and Q, X ¹⁴ can be selected from I and M, and X ¹⁵ can be: D, L, and N, X ¹⁶ can be V, X ¹⁷ can be G, X ¹⁸ can be selected from D and Q, X ¹⁹ can be selected from V, W, and Y, X ²⁰ can be W, X ²¹ can be P, and X ²² can be selected from D and S X ²³ can be selected from L and Q, and X ²⁴ can be selected from I, L, and V.

The IL-2Rβ ligand can comprise an amino acid sequence selected from any one of SEQ ID NO:167-SEQ ID NO:182.

The IL-2Rβ ligand can comprise an amino acid sequence selected from any one of SEQ ID NOS: 164-182, wherein the amino acid sequence is on the N-terminus, the C-terminus, or both the N-terminus and the C-terminus. It may end with the above amino acids -GG-.

The IL-2Rβ ligand may comprise an amino acid sequence selected from any one of SEQ ID NOs: 164-182, each amino acid independently having the following conservative substitutions: alanine (A), glycine (G), amino acids with small hydrophobic side chains including proline (P), serine (S), or threonine (T); hydroxyl-containing sides including serine (S), threonine (T), or tyrosine (Y) amino acids with acidic side chains, including aspartic acid (D), or glutamic acid (E); histidine (H), asparagine (N), glutamine (Q), serine (S), threonine (T), or amino acids with polar neutral side chains, including tyrosine (Y); amino acids with basic side chains, including arginine (R), lysine (K), or histidine (H); isoleucine (I), leucine (L) , methionine (M), valine (V), phenylalanine (F), tyrosine (Y), or tryptophan (W); and phenylalanine (F), histidine (H), tryptophan ( W), or amino acids with aromatic side chains, including tyrosine (Y).

The IL-2Rβ ligands of SEQ ID NOs:164-182 exhibit affinities for IL-2Rβ subunits of less than 100 μM.

The IL-2Rβ ligand can comprise the amino acid sequence of formula (2) (SEQ ID NO: 164), the amino acid sequence of formula (2a) (SEQ ID NO: 165), or the amino acid sequence of formula (2b) (SEQ ID NO: 166);
^{-X15 -X16 -X17 -X18 -X19 -X20 -X21 -X22} - (2)
^{-X14 -CX15 -X16 -X17 -X18 -X19 -X20 -X21 -X22 -CX23} - (2a)
^{-X13 -X14 -CX15 -X16 -X17 -X18 -X19 -X20 -X21 -X22 -CX23 -X24} - (2b)
wherein X ¹³ can be selected from amino acids, X ¹⁴ can be selected from amino acids containing large hydrophobic side chains, X ¹⁵ can be selected from amino acids, and X ¹⁶ can be selected from can be selected from amino acids containing large hydrophobic side chains, X ²⁷ can be selected from amino acids containing small hydrophobic side chains, X ¹⁸ can be selected from amino acids, and X ¹⁹ can be selected from can be selected from amino acids, X ²⁰ can be selected from amino acids containing large hydrophobic side chains, X ²¹ can be selected from amino acids containing small hydrophobic side chains, and X ²² can be selected from It can be selected from amino acids, ^X23 can be selected from amino acids, and ^X24 can be selected from amino acids containing large hydrophobic side chains.

In the IL-2Rβ ligands of formulas (2)-(2b), X ¹³ can be selected from amino acids, X ¹⁴ can be selected from amino acids containing large hydrophobic side chains, and X ¹⁵ can be selected from can be selected from amino acids, X ¹⁶ can be selected from amino acids containing large hydrophobic side chains, X ¹⁷ can be selected from amino acids containing small hydrophobic side chains, and X ¹⁸ can be selected from can be selected from amino acids containing polar neutral or acidic side chains, X ¹⁹ can be selected from amino acids containing large hydrophobic or neutral side chains, and X ²⁰ can be selected from amino acids containing large hydrophobic can be selected from amino acids containing side chains, X ²¹ can be selected from amino acids containing small hydrophobic side chains, X ²² can be selected from amino acids, X ²³ can be selected from amino acids and ^X24 can be selected from amino acids containing large hydrophobic side chains.

In the IL-2Rβ ligands of formulas (2)-(2b), X ¹³ can be selected from amino acids and X ¹⁴ can be selected from I, L, M, V, F, W, and Y. X ¹⁵ can be selected from D, E, I, L, M, V, F, Y, and W; X ¹⁶ can be selected from I, L, M, N, V, F, Y, and W, X ¹⁷ can be selected from A, G, P, S, and T, X ¹⁸ can be selected from H, N, Q, S, T, Y, D, and E X ¹⁹ can be selected from I, L, M, V, F, W, and Y; X ²⁰ can be selected from I, L, M, N, V, F, Y, and W, X ²¹ can be selected from A, G, P, S, and T, X ²² can be selected from amino acids, X ²³ can be selected from amino acids and X ²⁴ can be selected from I, L, M, V, F, W, and Y.

In the IL-2Rβ ligands of formulas (2)-(2b), ^X14 may be selected from I and M.

X ¹⁶ can be V in the IL-2Rβ ligands of formulas (2)-(2b).

X ¹⁷ can be G in the IL-2Rβ ligands of formulas (2)-(2b).

In IL-2Rβ ligands of formulas (2)-(2b), X ¹⁸ may be selected from D and Q.

X ²⁰ can be W in the IL-2Rβ ligands of formulas (2)-(2b).

In the IL-2Rβ ligands of formulas (2)-(2b), ^X21 can be P.

In the IL-2Rβ ligands of formulas (2)-(2b), ^X23 may be selected from F, I, L, and V.

In the IL-2Rβ ligands of formulas (2)-(2b), X ¹³ can be selected from amino acids, X ¹⁴ can be selected from I and M, and X ¹⁵ can be selected from amino acids. , X ¹⁶ can be V, X ¹⁷ can be G, X ¹⁸ can be selected from D and Q, X ¹⁹ can be I, L, M, V , F, W, and Y, X ²⁰ can be W, X ²¹ can be P, X ²² can be selected from amino acids, X ²³ can be selected from amino acids and ^X24 can be selected from F, I, L, and V.

The IL-2Rβ ligand can comprise the amino acid sequence of formula (3) (SEQ ID NO: 183) or the amino acid sequence of formula (3a) (SEQ ID NO: 184),
^{-X26 -X27 -X28 -X29 -X30 -X31 -X32 -X33 -X34 -X35} - ( ³ )
^{-X25 -CX26 -X27 -X28 -X29 -X30 -X31 -X32 -X33 -X34 -X35 -CX36} - (3a)
wherein X ²⁵ can be selected from amino acids, X ²⁶ can be selected from amino acids, X ²⁷ can be selected from I and V, X ²⁸ can be G X ²⁹ can be selected from D, E, and N, X ³⁰ can be selected from F, L, and Y, and X ³¹ can be selected from F, I, and V X ³² can be selected from D and Q, X ³³ can be selected from amino acids, X ³⁴ can be selected from amino acids, X ³⁵ can be selected from amino acids and X ³⁶ can be selected from amino acids.

In the IL-2Rβ ligands of formulas (3)-(3a), X ²⁵ can be selected from L, S, T, and Y, X ²⁶ can be selected from H and Q, X ²⁷ can be selected from I and V, X ²⁸ can be G, X ²⁹ can be selected from D, E, and N, X ³⁰ can be selected from F, L, and Y X ³¹ may be selected from F, I, and V; X ³² may be selected from D and Q; X ³³ may be selected from D, L, and W X ³⁴ can be selected from G, L, and T; X ³⁵ can be selected from D, I, and S; and X ³⁶ can be selected from A and M can be done.

The IL-2Rβ ligand can comprise the amino acid sequence of formula (3) (SEQ ID NO: 183) or the amino acid sequence of formula (3a) (SEQ ID NO: 184),
^{-X26 -X27 -X28 -X29 -X30} -X31 ^{-X32 -X33 -X34 -X35} - ( ³ )
^{-X25 -CX26 -X27 -X28 -X29 -X30 -X31 -X32 -X33 -X34 -X35 -CX36} - (3a)
wherein X ²⁵ can be selected from amino acids, X ²⁶ can be selected from amino acids, X ²⁷ can be selected from amino acids containing large hydrophobic side chains, and X ²⁸ can be selected from small hydrophobic side chains chain, X ²⁹ may be selected from amino acids containing an acidic side chain or a polar neutral side chain, X ³⁰ may be selected from amino acids, X ³¹ may be selected from amino acids X ³² can be selected from amino acids containing polar neutral or acidic side chains, X ³³ can be selected from amino acids, X ³⁴ can be selected from amino acids, X ³⁵ can be selected from amino acids and X ³⁶ can be selected from amino acids.

In the IL-2Rβ ligands of formulas (3)-(3a), X ²⁵ can be selected from amino acids, X ²⁶ can be selected from amino acids, and X ²⁷ contains a large hydrophobic side chain. can be selected from amino acids, X ²⁸ can be selected from amino acids containing small hydrophobic side chains, X ²⁹ can be selected from amino acids containing acidic or polar neutral side chains, ^X30 can be selected from amino acids containing large hydrophobic side chains, ^X31 can be selected from amino acids containing large hydrophobic side chains, and ^X32 can be selected from polar neutral side chains or acidic side chains. X ³³ can be selected from amino acids, X ³⁴ can be selected from amino acids, X ³⁵ can be selected from amino acids, X ³⁶ can be selected from , amino acids.

The IL-2Rβ ligand can comprise the amino acid sequence of formula (3) (SEQ ID NO: 183) or the amino acid sequence of formula (3a) (SEQ ID NO: 184),
^{-X26 -X27 -X28 -X29 -X30 -X31 -X32 -X33 -X34 -X35} - ( ³ )
^{-X25 -CX26 -X27 -X28 -X29 -X30 -X31 -X32 -X33 -X34 -X35 -CX36} - (3a)
wherein X ²⁵ can be selected from amino acids, X ²⁶ can be selected from amino acids, X ²⁷ can be selected from I, L, M, V, F, Y, and W X ²⁸ can be selected from A, G, P, S, and T; X ²⁹ can be selected from D, E, H, N, Q, S, T, and Y; X ³⁰ can be selected from I, L, M, V, F, Y, and W; X ³¹ can be selected from I, L, M, V, F, Y, and W; X ³² can be selected from D, E, H, N, Q, T, and Y; X ³³ can be selected from amino acids; X ³⁴ can be selected from amino acids; ³⁵ can be selected from amino acids and X ³⁶ can be selected from amino acids.

X ²⁷ may be selected from V and I in the IL-2Rβ ligands of formulas (3)-(3a).

In the IL-2Rβ ligands of formulas (3)-(3a), ^X28 can be G.

In the IL-2Rβ ligands of formulas (3)-(3a), ^X29 may be selected from D and E.

In IL-2Rβ ligands of formulas (3)-(3a), X ³⁰ may be selected from V, L, F, and Y.

In the IL-2Rβ ligands of formulas (3)-(3a), ^X31 may be selected from I, V, and F.

In the IL-2Rβ ligands of formulas (3)-(3a), ^X32 may be selected from Q and D.

In the IL-2Rβ ligands of formulas (3)-(3a), X ²⁵ can be selected from amino acids, X ²⁶ can be selected from amino acids, and X ²⁷ can be selected from V and I. , X ²⁸ can be G, X ²⁹ can be selected from D and E, X ³⁰ can be selected from V, L, F, and Y, and X ³¹ can be , I, V, and F, X ³² can be selected from Q and D, X ³³ can be selected from amino acids, X ³⁴ can be selected from amino acids X ³⁵ can be selected from amino acids and X ³⁶ can be selected from amino acids.

IL-2Rβ ligands may comprise an amino acid sequence selected from any one of SEQ ID NOs:185-193.

The IL-2Rβ ligand can comprise an amino acid sequence selected from any one of SEQ ID NOs: 183-193, wherein the amino acid sequence is on the N-terminus, the C-terminus, or both the N-terminus and the C-terminus. It may end with the above amino acids -GG-.

The IL-2Rβ ligand may comprise an amino acid sequence selected from any one of SEQ ID NOS: 183-193, each amino acid independently having the following conservative substitutions: alanine (A), glycine (G), amino acids with small hydrophobic side chains including proline (P), serine (S), or threonine (T); hydroxyl-containing sides including serine (S), threonine (T), or tyrosine (Y) amino acids with acidic side chains, including aspartic acid (D), or glutamic acid (E); histidine (H), asparagine (N), glutamine (Q), serine (S), threonine (T), or amino acids with polar neutral side chains, including tyrosine (Y); amino acids with basic side chains, including arginine (R), lysine (K), or histidine (H); isoleucine (I), leucine (L) , methionine (M), valine (V), phenylalanine (F), tyrosine (Y), or tryptophan (W); and phenylalanine (F), histidine (H), tryptophan ( W), or amino acids with aromatic side chains, including tyrosine (Y).

The IL-2Rβ ligands of SEQ ID NOs:183-193 exhibit affinities for IL-2Rβ subunits of less than 100 μM.

The IL-2Rβ ligand can comprise the amino acid sequence of formula (13) (SEQ ID NO: 1028),
-X ²⁰¹ -X ²⁰² -X ²⁰³ -X ²⁰⁴ -X ²⁰⁵ -X ²⁰⁶ -X ²⁰⁷ -X ²⁰⁸ -X ²⁰⁹ -X ²¹⁰ -X ²¹¹ -X ²¹² - (13)
wherein X ²⁰¹ can be selected from amino acids, X ²⁰² can be selected from amino acids, X ²⁰³ can be selected from amino acids containing acidic side chains, X ²⁰⁴ can be selected from large hydrophobic X ²⁰⁵ can be selected from amino acids containing small hydrophobic side chains, X ²⁰⁶ can be selected from amino acids containing acidic side chains, X ²⁰⁷ can be selected from amino acids containing large hydrophobic side chains, X ²⁰⁸ can be selected from amino acids, X ²⁰⁹ can be selected from amino acids containing acidic side chains, X ²¹⁰ can be selected from , amino acids, X ²¹¹ can be selected from amino acids, and X ²¹² can be selected from amino acids containing large hydrophobic side chains.

In the IL-2Rβ ligand of formula (13), X ²⁰¹ can be selected from amino acids, X ²⁰² can be selected from amino acids, X ²⁰³ can be selected from D and E, X ²⁰⁴ can be selected from I, L, M, V, F, Y, ^and W; X ²⁰⁵ can be selected from A, G, P, S, and T; and E, X ²⁰⁷ can be selected from I, L, M, V, F, Y, and W, X ²⁰⁸ can be selected from amino acids, and X ²⁰⁹ can be selected from D and E, X ²¹⁰ can be selected from amino acids, X ²¹¹ can be selected from amino acids, X ²¹² can be selected from I, L, M, V, F, Y, and W.

In the IL-2Rβ ligands of formula (13), X ²⁰¹ can be selected from C, F, L, S, and W, and X ²⁰² can be C, D, F, G, L, M, Q, S, V, W, and Y; X ²⁰³ may be selected from A, C, D, E, L, M, ^N , S, W, and Y; A, D, I, M, V, and W, X ²⁰⁵ can be selected from D, E, G, and I, X ²⁰⁶ can be selected from C, D, G, H, L, Q, S, and T; X ²⁰⁷ can be selected from C, D, I, L, V, W, and Y; X ²⁰⁸ can be selected from C, D, L, V, and W, X ²⁰⁹ can be selected from C, D, G, I, M, N, P, Q, and W, and X ²¹⁰ can be selected from D, F, L, M, P, S, T, and Y; X ²¹¹ can be selected from C, F, L, V, and W; X ²¹² can be selected from L, N, S, T, and V.

In IL-2Rβ ligands of formula (13), X ²⁰¹ may be selected from C, F, L, S, and W.

In IL-2Rβ ligands of formula (13), X ²⁰² may be selected from C, D, F, G, L, M, Q, S, V, W, and Y.

In IL-2Rβ ligands of formula (13), ^X203 may be selected from D and E.

X ²⁰⁴ can be V in the IL-2Rβ ligand of formula (13).

X ²⁰⁵ can be G in the IL-2Rβ ligand of formula (13).

X ²⁰⁶ can be D in the IL-2Rβ ligand of formula (13).

X ²⁰⁷ may be selected from I, W, and Y in the IL-2Rβ ligand of formula (13).

X ²⁰⁸ may be selected from C, D, L, V, and W in IL-2Rβ ligands of formula (13).

X ²⁰⁹ can be D in the IL-2Rβ ligand of formula (13).

In IL-2Rβ ligands of formula (13), X ²¹⁰ may be selected from D, F, L, M, P, S, T, and Y.

In IL-2Rβ ligands of formula (13), X ²¹¹ may be selected from C, F, L, V, and W.

In IL-2Rβ ligands of formula (13), X ²¹² may be selected from L and V.

In the IL-2Rβ ligand of formula (13), X ²⁰¹ can be selected from amino acids, X ²⁰² can be selected from amino acids, X ²⁰³ can be selected from D and E, X ²⁰⁴ can be V, X ²⁰⁵ can be G, X ²⁰⁶ can be D, X ²⁰⁷ can be selected from I, Y, and W; ²⁰⁸ can be selected from amino acids, X ²⁰⁹ can be D, X ²¹⁰ can be selected from amino acids, X ²¹¹ can be selected from amino acids, X ²¹² can be selected from I, L, M, V, F, Y, and W can be selected.

The IL-2Rβ ligand can comprise an amino acid sequence selected from any one of SEQ ID NOs:1029-1043.

The IL-2Rβ ligand can comprise an amino acid sequence selected from any one of SEQ ID NOs: 1028-1043, wherein the amino acid sequence is on the N-terminus, the C-terminus, or both the N-terminus and the C-terminus. It may end with the above amino acids -GG-.

The IL-2Rβ ligand may comprise an amino acid sequence selected from any one of SEQ ID NOS: 1028-1043, each amino acid independently having the following conservative substitutions: alanine (A), glycine (G), amino acids with small hydrophobic side chains including proline (P), serine (S), or threonine (T); hydroxyl-containing sides including serine (S), threonine (T), or tyrosine (Y) amino acids with acidic side chains, including aspartic acid (D), or glutamic acid (E); histidine (H), asparagine (N), glutamine (Q), serine (S), threonine (T), or amino acids with polar neutral side chains, including tyrosine (Y); amino acids with basic side chains, including arginine (R), lysine (K), or histidine (H); isoleucine (I), leucine (L) , methionine (M), valine (V), phenylalanine (F), tyrosine (Y), or tryptophan (W); and phenylalanine (F), histidine (H), tryptophan ( W), or amino acids with aromatic side chains, including tyrosine (Y).

The IL-2β ligand can comprise an amino acid sequence selected from any one of SEQ ID NOS:1044-1050.

The IL-2Rβ ligand can comprise an amino acid sequence selected from any one of SEQ ID NOs: 1044-1050, wherein the amino acid sequence is on the N-terminus, the C-terminus, or both the N-terminus and the C-terminus. It may end with the above amino acids -GG-.
The IL-2Rβ ligand may comprise an amino acid sequence selected from any one of SEQ ID NOs: 1044-1050, each amino acid independently having the following conservative substitutions: alanine (A), glycine (G), amino acids with small hydrophobic side chains including proline (P), serine (S), or threonine (T); hydroxyl-containing sides including serine (S), threonine (T), or tyrosine (Y) amino acids with acidic side chains, including aspartic acid (D), or glutamic acid (E); histidine (H), asparagine (N), glutamine (Q), serine (S), threonine (T), or amino acids with polar neutral side chains, including tyrosine (Y); amino acids with basic side chains, including arginine (R), lysine (K), or histidine (H); isoleucine (I), leucine (L) , methionine (M), valine (V), phenylalanine (F), tyrosine (Y), or tryptophan (W); and phenylalanine (F), histidine (H), tryptophan ( W), or amino acids with aromatic side chains, including tyrosine (Y).
IL-2Rβ ligands of SEQ ID NOs:144-150 exhibit affinities for IL-2Rβ subunits of less than 100 μM.

The IL-2β ligand can comprise the amino acid sequence of formula (10) (SEQ ID NO:578),
-X ¹⁹¹ -X ¹⁹² -X ¹⁹³ -X ¹⁹⁴ -X ¹⁹⁵ -CX ^{196 -X} 197 ^-X 198 -X ¹⁹⁹ -X ²⁰⁰ -X ²⁰¹ -X ²⁰² -X ²⁰³ -CX ²⁰⁴ -X ²⁰⁵ - X ²⁰⁶ -X ²⁰⁷ -X ²⁰⁸ - (10)
wherein X ¹⁹¹ can be selected from amino acids containing large hydrophobic or aromatic side chains, X ¹⁹² can be selected from amino acids, and X ¹⁹³ can be selected from large hydrophobic or aromatic side chains. X ¹⁹⁴ can be selected from amino acids containing a large hydrophobic side chain or a basic side chain, X ¹⁹⁵ can be selected from amino acids containing an acidic side chain or a small hydrophobic side chain X ¹⁹⁶ can be selected from amino acids containing large hydrophobic or basic side chains, X ¹⁹⁷ can be selected from amino acids containing large hydrophobic side chains X ¹⁹⁸ can be selected from amino acids containing small hydrophobic side chains, X ¹⁹⁹ can be selected from amino acids containing polar/neutral side chains or basic side chains, and X ²⁰⁰ can be , can be selected from amino acids containing large hydrophobic side chains, X ²⁰¹ can be selected from amino acids containing small hydrophobic side chains, and X ²⁰² can be selected from amino acids containing acidic or polar/neutral side chains. X ²⁰³ can be selected from amino acids containing a large hydrophobic side chain, X ²⁰⁴ can be selected from amino acids containing an acidic side chain, X ²⁰⁵ can be selected from amino acids containing a large can be selected from amino acids containing hydrophobic side chains, X ²⁰⁶ can be selected from amino acids containing acidic or aromatic side chains, and X ²⁰⁷ can be selected from amino acids containing amino acids. , X ²⁰⁸ can be selected from amino acids containing acidic side chains.

In the IL-2Rβ ligands of formula (10), X ¹⁹¹ can be selected from F, H, I, L, M, V, W, and Y, X ¹⁹² can be selected from amino acids, X ¹⁹³ can be selected from F, H, I, L, V, W, and Y; X ¹⁹⁴ can be selected from F, I, L, M, V, W, Y, H, K, and R; X ¹⁹⁵ can be selected from D, E, A, G, P, S, and T; X ¹⁹⁶ can be selected from F, I, L, M, V, W, Y, H , K, and R, X ¹⁹⁷ can be selected from F, I, L, M, V, W, and Y, and X ¹⁹⁸ can be selected from A, G, P, S, and T, X ¹⁹⁹ can be selected from H, N, Q, S, T, Y, H, K, and R; X ²⁰⁰ can be selected from F, I, L, M, V , W, and Y, X ²⁰¹ can be selected from A, G, P, S, and T, and X ²⁰² can be selected from D, E, H, N, Q, S, T , and Y, X ²⁰³ may be selected from F, I, L, M, V, W, and Y, X ²⁰⁴ may be selected from D and E, and X ²⁰⁵ can be selected from F, I, L, M, V, W, and Y, and X ²⁰⁶ can be selected from D, E, F, H, I, L, M, V, W, and Y X ²⁰⁷ can be selected from amino acids and X ²⁰⁸ can be selected from D and E.

In IL-2Rβ ligands of formula (10), X ¹⁹¹ may be selected from F, H, W, and Y.

X ¹⁹¹ can be W in the IL-2Rβ ligand of formula (10).

In IL-2Rβ ligands of formula (10), ^X192 may be selected from amino acids.

X ¹⁹³ may be selected from F, H, W, and Y in the IL-2Rβ ligand of formula (10).

X ¹⁹³ may be selected from F, W, and Y in the IL-2Rβ ligand of formula (10).

X ¹⁹⁴ may be selected from H, L, and Y in the IL-2Rβ ligands of formula (10).

X ¹⁹⁴ can be L in the IL-2Rβ ligand of formula (10).

X ¹⁹⁴ can be Y in the IL-2Rβ ligand of formula (10).

In IL-2Rβ ligands of formula (10), ^X195 may be selected from D and P.

X ¹⁹⁵ can be D in the IL-2Rβ ligand of formula (10).

X ¹⁹⁵ can be P in the IL-2Rβ ligand of formula (10).

X ¹⁹⁶ may be selected from H and W in the IL-2Rβ ligand of formula (10).

X ¹⁹⁶ can be H in IL-2Rβ ligands of formula (10).

X ¹⁹⁶ can be W in the IL-2Rβ ligand of formula (10).

X ¹⁹⁷ can be M in the IL-2Rβ ligand of formula (10).

X ¹⁹⁸ can be A in the IL-2Rβ ligand of formula (10).

In IL-2Rβ ligands of formula (10), X ¹⁹⁹ may be selected from H, K, R, and Q.

X ¹⁹⁹ can be Q in the IL-2Rβ ligand of formula (10).

In IL-2Rβ ligands of formula (10), X ¹⁹⁹ may be selected from H, K, and R.

In IL-2Rβ ligands of formula (10), ^X200 may be selected from L and V.

X ²⁰⁰ can be L in the IL-2Rβ ligand of formula (10).

X ²⁰¹ can be G in the IL-2Rβ ligand of formula (10).

In IL-2Rβ ligands of formula (10), X ²⁰² may be selected from D, E, and Q.

X ²⁰² can be E in the IL-2Rβ ligand of formula (10).

X ²⁰³ can be L in the IL-2Rβ ligand of formula (10).

In IL-2Rβ ligands of formula (10), ^X204 may be selected from D and E.

X ²⁰⁴ can be D in the IL-2Rβ ligand of formula (10).

X ²⁰⁵ can be L in the IL-2Rβ ligand of formula (10).

In IL-2Rβ ligands of formula (10), X ²⁰⁶ may be selected from D and E.

In IL-2Rβ ligands of formula (10), X ²⁰⁷ may be selected from amino acids.

In IL-2Rβ ligands of formula (10), X ²⁰⁸ may be selected from D and E.

In the IL-2Rβ ligands of formula (10), X ¹⁹¹ can be selected from F, H, W, and Y, X ¹⁹² can be selected from amino acids, X ¹⁹³ can be F, H, W, and Y; X ¹⁹⁴ can be selected from H, L, ^and Y; X ¹⁹⁵ can be selected from D and P; and W, X ¹⁹⁷ can be M, X ¹⁹⁸ can be A, X ¹⁹⁹ can be selected from H, K, R, and Q; X ²⁰⁰ can be selected from L and V, X ²⁰¹ can be G, X ²⁰² can be selected from D, E, and Q, and X ²⁰³ can be L , X ²⁰⁴ can be selected from D and E, X ²⁰⁵ can be L, and X ²⁰⁶ can be selected from D, E, H, F, W, and Y , X ²⁰⁷ can be selected from amino acids and X ²⁰⁸ can be selected from D and E.

In the IL-2Rβ ligand of formula (10), X ¹⁹¹ can be selected from F, H, W, and Y, X ¹⁹² can be selected from amino acids, and X ¹⁹³ is Y and X ¹⁹⁴ can be selected from H, L, and Y, X ¹⁹⁵ can be D, X ¹⁹⁶ can be W, and X ¹⁹⁷ can be M can be, X ¹⁹⁸ can be A, X ¹⁹⁹ can be Q, X ²⁰⁰ can be selected from L and V, X ²⁰¹ can be G, X ²⁰² can be selected from D, E, and Q, X ²⁰³ can be L, X ²⁰⁴ can be selected from D and E, and X ²⁰⁵ can be L X ²⁰⁶ can be selected from D and E, X ²⁰⁷ can be selected from amino acids, and X ²⁰⁸ can be selected from D and E.

In the IL-2Rβ ligand of formula (10), X ¹⁹¹ can be selected from F, H, W, and Y, X ¹⁹² can be selected from amino acids, and X ¹⁹³ is Y and X ¹⁹⁴ can be selected from H, L, and Y, X ¹⁹⁵ can be D, X ¹⁹⁶ can be H, and X ¹⁹⁷ can be M can be, X ¹⁹⁸ can be A, X ¹⁹⁹ can be Q, X ²⁰⁰ can be selected from L and V, X ²⁰¹ can be G, X ²⁰² can be selected from D, E, and Q, X ²⁰³ can be L, X ²⁰⁴ can be selected from D and E, and X ²⁰⁵ can be L X ²⁰⁶ can be selected from D and E, X ²⁰⁷ can be selected from amino acids, and X ²⁰⁸ can be selected from D and E.

In the IL-2Rβ ligand of formula (10), X ¹⁹¹ can be selected from F, H, W, and Y, X ¹⁹² can be selected from amino acids, and X ¹⁹³ is Y and X ¹⁹⁴ can be selected from H, L, and Y, X ¹⁹⁵ can be D, X ¹⁹⁶ can be R, and X ¹⁹⁷ can be M can be, X ¹⁹⁸ can be A, X ¹⁹⁹ can be Q, X ²⁰⁰ can be selected from L and V, X ²⁰¹ can be G, X ²⁰² can be selected from D, E, and Q, X ²⁰³ can be L, X ²⁰⁴ can be selected from D and E, and X ²⁰⁵ can be L X ²⁰⁶ can be selected from D and E, X ²⁰⁷ can be selected from amino acids, and X ²⁰⁸ can be selected from D and E.

In the IL-2Rβ ligand of formula (10), X ¹⁹¹ can be selected from F, H, W, and Y, X ¹⁹² can be selected from amino acids, and X ¹⁹³ is Y and X ¹⁹⁴ can be selected from H, L, and Y, X ¹⁹⁵ can be P, X ¹⁹⁶ can be W, and X ¹⁹⁷ can be M can be, X ¹⁹⁸ can be A, X ¹⁹⁹ can be Q, X ²⁰⁰ can be selected from L and V, X ²⁰¹ can be G, X ²⁰² can be selected from D, E, and Q, X ²⁰³ can be L, X ²⁰⁴ can be selected from D and E, and X ²⁰⁵ can be L X ²⁰⁶ can be selected from D and E, X ²⁰⁷ can be selected from amino acids, and X ²⁰⁸ can be selected from D and E.

In the IL-2Rβ ligand of formula (10), X ¹⁹¹ can be selected from F, H, W, and Y, X ¹⁹² can be selected from amino acids, and X ¹⁹³ is Y and X ¹⁹⁴ can be selected from H, L, and Y, X ¹⁹⁵ can be D, X ¹⁹⁶ can be W, and X ¹⁹⁷ can be M , X ¹⁹⁸ can be A, X ¹⁹⁹ can be selected from H, K, and R, X ²⁰⁰ can be selected from L and V, X ²⁰¹ can be G , X ²⁰² can be selected from D, E, and Q, X ²⁰³ can be L, X ²⁰⁴ can be selected from D and E, X ²⁰⁵ can be L, X ²⁰⁶ can be selected from D and E, X ²⁰⁷ can be selected from amino acids, and X ²⁰⁸ can be selected from D and E.

In the IL-2Rβ ligand of formula (10), X ¹⁹¹ can be selected from F, H, W, and Y, X ¹⁹² can be selected from amino acids, and X ¹⁹³ is Y and X ¹⁹⁴ can be selected from H, L, and Y, X ¹⁹⁵ can be D, X ¹⁹⁶ can be W, and X ¹⁹⁷ can be M can be, X ¹⁹⁸ can be A, X ¹⁹⁹ can be Q, X ²⁰⁰ can be selected from L and V, X ²⁰¹ can be G, X ²⁰² can be selected from D, E, and Q, X ²⁰³ can be L, X ²⁰⁴ can be selected from D and E, and X ²⁰⁵ can be L X ²⁰⁶ can be selected from F, H, W, and Y, X ²⁰⁷ can be selected from amino acids, and X ²⁰⁸ can be selected from D and E.

In the IL-2Rβ ligand of formula (10), X ¹⁹¹ is A, D, E, F, G, H, I, K, L, N, M, P, Q, R, S, T, V, W , and Y, and X ¹⁹² can be selected from A, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, and Y, and X ¹⁹³ may be selected from A, C, D, F, G, H, I, L, M, NP, R, S, T, V, W, and Y and X ¹⁹⁴ can be selected from F, H, I, K, L, N, P, Q, R, S, T, V, W, and Y, and X ¹⁹⁵ can be selected from A, D, E, F, G, H, K, L, M, N, P, Q, S, W, and Y, where X ¹⁹⁶ is A, E, F, G, H, Q, R , S, W, and Y, and X ¹⁹⁷ can be selected from A, D, E, F, I, K, L, M, N, Q, R, S, T, V, W, and Y and X ¹⁹⁸ can be A and X ¹⁹⁹ can be selected from A, D, H, K, L, N, P, Q, R, S, and Y , X ²⁰⁰ can be selected from I, L, M, P, and V, X ²⁰¹ can be selected from G, H, and W, and X ²⁰² can be selected from D, E, and Q X ²⁰³ can be L, X ²⁰⁴ can be selected from A, D, E, H, I, L, T, V, and Y, and X ²⁰⁵ can be: F, I, L, M, V, W, and Y, and X ²⁰⁶ is A, D, E, F, G, H, I, K, L, M, N, P, Q , R, S, T, V, W, and Y, and X ²⁰⁷ can be selected from A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, and Y, and X ²⁰⁸ is A, D, E, F, G, H, K, L, M, N, P, Q, R, S , T, V, W, and Y.

In the IL-2Rβ ligands of formula (10), X ¹⁹¹ can be selected from F, H, W, and Y, and X ¹⁹² can be A, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, and Y; X ¹⁹³ can be selected from F ^, H, W, and Y; can be selected from F, H, I, L, V, W, and Y, X ¹⁹⁵ can be selected from D, E, and P, and X ¹⁹⁶ can be selected from F, H, R, S, W, and Y, X ¹⁹⁷ can be selected from F, I, L, M, and V, X ¹⁹⁸ can be A, X ¹⁹⁹ can be H, K , N, Q, and R, X ²⁰⁰ can be selected from I, L, and V, X ²⁰¹ can be G, and X ²⁰² can be from D, E, and Q X ²⁰³ can be selected from F, I, L, M, V, and Y, X ²⁰⁴ can be selected from D and E, and X ²⁰⁵ can be L , X ²⁰⁶ can be selected from D, E, N, and Q, and X ²⁰⁷ can be selected from A, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, and Y can be selected, and X ²⁰⁸ can be selected from D and E.

In the IL-2Rβ ligand of formula (10), X ¹⁹¹ can be W and X ¹⁹² are A, D, E, F, G, H, I, K, L, M, N, P, Q , R, S, T, V, W, and Y; X ¹⁹³ may be selected from F, H, W, and Y; X ¹⁹⁴ may be Y; ¹⁹⁵ can be selected from D, E, and P; X ¹⁹⁶ can be selected from H, R, ^and W; X ¹⁹⁷ can be selected from I and M; A, X ¹⁹⁹ can be selected from K, Q, and R, X ²⁰⁰ can be selected from I, L, and V, and X ²⁰¹ can be G X ²⁰² can be E, X ²⁰³ can be L, X ²⁰⁴ can be D, X ²⁰⁵ can be L, X ²⁰⁶ can be from D and E and X ²⁰⁷ is selected from A, D, E, F, G, H, I, L, M, N, Q, P, R, S, T, V, W, and Y and X ²⁰⁸ can be selected from D and E.

In the IL-2Rβ ligand of formula (10), X ¹⁹¹ is A, D, E, F, G, H, I, K, L, N, M, P, Q, R, S, T, V, W , and Y, and X ¹⁹² can be selected from A, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, and Y; X ¹⁹³ may be selected from F, H, W, and Y; X ¹⁹⁴ may be selected from F, H, L, W, and Y; ¹⁹⁵ can be selected from D, E, and P; X ¹⁹⁶ can be selected from F, H, R, S, W, and Y; X ¹⁹⁷ can be selected from F, I, L, M , and V, X ¹⁹⁸ can be A, X ¹⁹⁹ can be selected from H, K, Q, N, and R, X ²⁰⁰ can be I, L, and V, X ²⁰¹ can be G, X ²⁰² can be selected from D, E, and Q, X ²⁰³ can be L, X ²⁰⁴ can be selected from D and E, X ²⁰⁵ can be selected from F, I, L, M, V, and W, and X ²⁰⁶ can be selected from D, E, F, I, L, M , V, W, and Y. X ²⁰⁷ can be selected from A, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, and Y ^; can be selected from D and E.

In the IL-2Rβ ligand of formula (10), X ¹⁹¹ is A, D, E, F, G, H, I, K, L, N, M, P, Q, R, S, T, V, W , and Y.

In IL-2Rβ ligands of formula (10), X ¹⁹¹ may be selected from A, G, P, S, and T.

In IL-2Rβ ligands of formula (10), X ¹⁹¹ may be selected from F, H, I, L, M, V, W, and Y.

In IL-2Rβ ligands of formula (10), X ¹⁹¹ may be selected from F, H, W, and Y.

In the IL-2Rβ ligand of formula (10), X ¹⁹² is A, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W , and Y.

In IL-2Rβ ligands of formula (10), X ¹⁹² may be selected from A, G, P, S, and T.

In IL-2Rβ ligands of formula (10), X ¹⁹² may be selected from F, H, I, L, M, V, W, and Y.

X ¹⁹³ may be selected from F, H, W, and Y in the IL-2Rβ ligand of formula (10).

X ¹⁹³ can be W in the IL-2Rβ ligand of formula (10).

In IL-2Rβ ligands of formula (10), X ¹⁹⁴ may be selected from F, H, L, W, and Y.

X ¹⁹⁴ may be selected from H, L, and Y in the IL-2Rβ ligands of formula (10).

X ¹⁹⁴ can be Y in the IL-2Rβ ligand of formula (10).

In IL-2Rβ ligands of formula (10), ^X195 may be selected from D, E, and P.

X ¹⁹⁵ can be D in the IL-2Rβ ligand of formula (10).

X ¹⁹⁵ can be P in the IL-2Rβ ligand of formula (10).

In IL-2Rβ ligands of formula (10), X ¹⁹⁶ may be selected from F, H, R, S, W, and Y.

X ¹⁹⁶ may be selected from H, R, and W in the IL-2Rβ ligand of formula (10).

X ¹⁹⁶ can be W in the IL-2Rβ ligand of formula (10).

In IL-2Rβ ligands of formula (10), X ¹⁹⁷ may be selected from F, I, L, M, and V.

In IL-2Rβ ligands of formula (10), X ¹⁹⁷ may be selected from I and M.

X ¹⁹⁷ can be M in the IL-2Rβ ligand of formula (10).

X ¹⁹⁸ can be A in the IL-2Rβ ligand of formula (10).

In IL-2Rβ ligands of formula (10), ^X199 may be selected from H, K, Q, N, and R.

In IL-2Rβ ligands of formula (10), X ¹⁹⁹ may be selected from H, K, and R.

X ¹⁹⁹ can be Q in the IL-2Rβ ligand of formula (10).

In IL-2Rβ ligands of formula (10), ^X200 may be selected from I, L, and V.

In IL-2Rβ ligands of formula (10), ^X200 may be selected from L and V.

X ²⁰¹ can be G in the IL-2Rβ ligand of formula (10).

In IL-2Rβ ligands of formula (10), X ²⁰² may be selected from D, E, and Q.

X ²⁰² can be E in the IL-2Rβ ligand of formula (10).

X ²⁰³ can be L in the IL-2Rβ ligand of formula (10).

In IL-2Rβ ligands of formula (10), ^X204 may be selected from D and E.

X ²⁰⁴ can be D in the IL-2Rβ ligand of formula (10).

In IL-2Rβ ligands of formula (10), X ²⁰⁵ may be selected from F, I, L, M, V, and W.

X ²⁰⁵ can be L in the IL-2Rβ ligand of formula (10).

In IL-2Rβ ligands of formula (10), X ²⁰⁶ may be selected from D and E.

X ²⁰⁶ can be D in the IL-2Rβ ligand of formula (10).

In IL-2Rβ ligands of formula (10), X ²⁰⁶ may be selected from F, I, L, M, V, W, and Y.

In the IL-2Rβ ligand of formula (10), X ²⁰⁷ is A, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, and can be selected from Y;

In IL-2Rβ ligands of formula (10), X ²⁰⁷ may be selected from A, G, P, S, and T.

In IL-2Rβ ligands of formula (10), X ²⁰⁷ may be selected from F, I, L, M, V, W, and Y.

In IL-2Rβ ligands of formula (10), X ²⁰⁸ may be selected from D and E.

In the IL-2Rβ ligands of formula (10), X ¹⁹¹ can be selected from F, I, L, M, V, W, and Y, X ¹⁹² can be selected from A, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, and Y, and X ¹⁹³ is selected from F, H, W, and Y X ¹⁹⁴ can be selected from H, L, and Y, X ¹⁹⁵ can be selected from D and P, and X ¹⁹⁶ can be selected from H, R, and W X ¹⁹⁷ can be selected from I and M; X ¹⁹⁸ can be A; X ¹⁹⁹ can be selected from H ^, K, Q, and R; L and V, X ²⁰¹ can be G, X ²⁰² can be selected from D, E, and Q, X ²⁰³ can be L, X ²⁰⁴ can be , D and E, X ²⁰⁵ can be L, X ²⁰⁶ can be selected from D, E, F, I, L, M, V, W, and Y; X ²⁰⁷ can be selected from A, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, and Y, and X ²⁰⁸ is , D and E.

In the IL-2Rβ ligands of formula (10), X ¹⁹¹ can be selected from F, I, L, M, V, W, and Y, X ¹⁹² can be selected from A, D, E, F, G, can be selected from H, I, K, L, M, N, P, Q ^, R, S, T, V, W, and Y; X ¹⁹³ can be W; Y, X ¹⁹⁵ can be selected from D and P, X ¹⁹⁶ can be W, X ¹⁹⁷ can be M, X ¹⁹⁸ can be A X ¹⁹⁹ can be Q, X ¹⁹⁹ can be selected from H, K, and R, X ²⁰⁰ can be selected from L and V, and X ²⁰¹ can be can be G, X ²⁰² can be E, X ²⁰³ can be L, X ²⁰⁴ can be selected from D and E, X ²⁰⁵ is L X ²⁰⁶ can be D and X ²⁰⁷ can be A, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V , and W, and Y, and X ²⁰⁸ can be selected from D and E.

In IL-2Rβ ligands of formula (10), X ¹⁹¹ can be selected from F, I, L, M, V, W, and Y, X ¹⁹³ can be W, and X ¹⁹⁴ can be , Y, X ¹⁹⁵ can be selected from D and P, X ¹⁹⁶ can be selected from H, R, and W, X ¹⁹⁷ can be M, X ¹⁹⁸ can be A, X ¹⁹⁹ can be selected from H, K, Q, and R, X ²⁰⁰ can be selected from L and V, X ²⁰¹ can be G X ²⁰² can be E, X ²⁰³ can be L, X ²⁰⁴ can be D, X ²⁰⁵ can be L, X ²⁰⁶ can be D and X ²⁰⁷ can be A, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, and Y and X ²⁰⁸ can be selected from D and E.

IL-2Rβ ligands may comprise an amino acid sequence selected from any one of SEQ ID NO:579-SEQ ID NO:808.

The IL-2Rβ ligand can comprise an amino acid sequence selected from any one of SEQ ID NOs:578-808, wherein the amino acid sequence is on the N-terminus, the C-terminus, or both the N-terminus and the C-terminus. It may end with the above amino acids -GG-.

The IL-2Rβ ligand may comprise an amino acid sequence selected from any one of SEQ ID NOs:578-808, each amino acid independently having the following conservative substitutions: alanine (A), glycine (G), amino acids with small hydrophobic side chains including proline (P), serine (S), or threonine (T); hydroxyl-containing sides including serine (S), threonine (T), or tyrosine (Y) amino acids with acidic side chains, including aspartic acid (D), or glutamic acid (E); histidine (H), asparagine (N), glutamine (Q), serine (S), threonine (T), or amino acids with polar neutral side chains, including tyrosine (Y); amino acids with basic side chains, including arginine (R), lysine (K), or histidine (H); isoleucine (I), leucine (L) , methionine (M), valine (V), phenylalanine (F), tyrosine (Y), or tryptophan (W); and phenylalanine (F), histidine (H), tryptophan ( W), or amino acids with aromatic side chains, including tyrosine (Y).

The IL-2Rβ ligands of SEQ ID NO:578-SEQ ID NO:808 exhibit affinities for IL-2Rβ subunits of less than 100 μM.

The IL-2β ligand can comprise the amino acid sequence of formula (11) (SEQ ID NO:809),
-X ²¹¹ -X ²¹² -X ²¹³ -X ²¹⁴ -CX ²¹⁵ -X ^{216 -X} 217 -X 218 ^{-X 219} -X ²²⁰ -X ²²¹ -X ²²² -CX ²²³ -X ²²⁴ -X ²²⁵ - (11)
wherein X ²¹¹ can be selected from amino acids, X ²¹² can be selected from amino acids containing aromatic side chains, X ²¹³ can be selected from amino acids containing large hydrophobic or aromatic side chains X ²¹⁴ can be P, X ²¹⁵ can be selected from amino acids containing aromatic side chains, X ²¹⁶ can be selected from amino acids containing large hydrophobic side chains X ²¹⁷ can be A, X ²¹⁸ can be selected from amino acids containing basic or polar/neutral side chains, X ²¹⁹ can be a large hydrophobic side chain X ²²⁰ can be G, X ²²¹ can be selected from amino acids containing an acidic or polar/neutral side chain, X ²²² can be selected from L X ²²³ can be D, X ²²⁴ can be selected from amino acids containing large hydrophobic side chains, and X ²²⁵ can be selected from amino acids containing acidic side chains can.

In the IL-2Rβ ligands of formula (11), X ²¹¹ can be selected from amino acids, X ²¹² can be selected from F, H, W and Y, X ²¹³ can be selected from F, H, X ²¹⁴ can be selected from I, L, M, V, W, and Y; X ²¹⁵ can be selected from F, H, W, and Y; ²¹⁶ can be selected from F, I, L, M, V, W, and Y, X ²¹⁷ can be A, X ²¹⁸ can be K, R, H, N, Q, S , T, and Y, X ²¹⁹ can be selected from F, I, L, M, V, W, and Y, X ²²⁰ can be G, and X ²²¹ can be selected from D, E, H, N, Q, S, T, and Y, X ²²² can be L, X ²²³ can be D, and X ²²⁴ can be , F, I, L, M, V, W, and Y, and X ²²⁵ can be selected from D and E.

In IL-2Rβ ligands of formula (11), X ²¹¹ may be selected from amino acids.

In IL-2Rβ ligands of formula (11), X ²¹¹ may be selected from H, K, and R.

In IL-2Rβ ligands of formula (11), X ²¹¹ may be selected from H and R.

In IL-2Rβ ligands of formula (11), X ²¹² may be selected from F, H, W, and Y.

X ²¹² can be W in the IL-2Rβ ligand of formula (11).

In IL-2Rβ ligands of formula (11), X ²¹³ may be selected from F, H, I, L, M, V, W, and Y.

X ²¹³ can be L in the IL-2Rβ ligand of formula (11).

X ²¹³ can be Y in the IL-2Rβ ligand of formula (11).

X ²¹⁴ can be P in the IL-2Rβ ligand of formula (11).

In IL-2Rβ ligands of formula (11), X ²¹⁵ may be selected from F, H, W, and Y.

X ²¹⁵ can be W in the IL-2Rβ ligand of formula (11).

In IL-2Rβ ligands of formula (11), X ²¹⁶ may be selected from F, I, L, M, V, W, and Y.

X ²¹⁶ can be M in IL-2Rβ ligands of formula (11).

X ²¹⁷ can be A in the IL-2Rβ ligand of formula (11).

In IL-2Rβ ligands of formula (11), X ²¹⁸ may be selected from K, R, H, N, Q, S, T, and Y.

In IL-2Rβ ligands of formula (11), X ²¹⁸ may be selected from K and R.

X ²¹⁸ can be Q in the IL-2Rβ ligand of formula (11).

In IL-2Rβ ligands of formula (11), X ²¹⁹ may be selected from F, I, L, M, V, W, and Y.

X ²¹⁹ can be L in the IL-2Rβ ligand of formula (11).

X ²²⁰ can be G in the IL-2Rβ ligand of formula (11).

In IL-2Rβ ligands of formula (11), X ²²¹ may be selected from D, E, H, N, Q, S, T, and Y.

X ²²¹ can be E in the IL-2Rβ ligand of formula (11).

X ²²² can be L in the IL-2Rβ ligand of formula (11).

X ²²³ can be D in the IL-2Rβ ligand of formula (11).

In IL-2Rβ ligands of formula (11), X ²²⁴ may be selected from F, I, L, M, V, W, and Y.

X ²²⁴ can be L in the IL-2Rβ ligand of formula (11).

In IL-2Rβ ligands of formula (11), X ²²⁵ may be selected from D and E.

In IL-2Rβ ligands of formula (11), X ²¹¹ can be selected from H, K, and R, X ²¹² can be W, X ²¹³ can be Y, X ²¹⁴ can be P, X ²¹⁵ can be W, X ²¹⁶ can be M, X ²¹⁷ can be A, X ²¹⁸ can be N and Q, X ²¹⁹ can be selected from L and V, X ²²⁰ can be G, X ²²¹ can be selected from E, D, and Q; X ²²² can be L, X ²²³ can be D, X ²²⁴ can be selected from L and M, and X ²²⁵ can be selected from D and E.

In the IL-2Rβ ligand of formula (11), X ²¹¹ can be selected from A, D, E, G, H, L, M, N, Q, R, S, T, and ^V ; can be selected from C, F, W, and Y; X ²¹³ can be selected ^from F, H, K, L, N, Q, R, S, W, and Y; can be P, X ²¹⁵ can be selected from W and Y, and X ²¹⁶ can be selected from F, I, K, L, M, R, S, T, and V X ²¹⁷ can be A, X ²¹⁸ can be selected from D, E, G, H, K, L, N, Q, R, S, and Y, and X ²¹⁹ can be L, P, and V can be selected, X ²²⁰ can be selected from G, H, and W, X ²²¹ can be selected from D, E, and Q, and X ²²² can be selected from , L and M, X ²²³ can be D, X ²²⁴ can be selected from L, M, Q, and V, and X ²²⁵ can be A, D, E , F, G, H, L, Q, N, T, and V.

In the IL-2Rβ ligands of formula (11), X ²¹¹ can be selected from H and R, X ²¹² can be selected from F and W, X ²¹³ can be F, L, W, and Y, X ²¹⁴ can be P, X ²¹⁵ can be selected from W and Y, X ²¹⁶ can be selected from F, I, L, M, and V X ²¹⁷ can be A, X ²¹⁸ can be selected from D, E, H, K, N, Q, and R, and X ²¹⁹ can be selected from L and V ^{X 220} can be G; X ²²¹ can be selected from D, E, and Q; X ²²² can be selected from L and M; D, X ²²⁴ can be selected from L, M, and V, and X ²²⁵ can be selected from D and E.

In IL-2Rβ ligands of formula (11), X ²¹¹ can be selected from H and R, X ²¹² can be W, X ²¹³ can be Y and X ²¹⁴ is P X ²¹⁵ can be W, X ²¹⁶ can be M, X ²¹⁷ can be A, X ²¹⁸ can be Q, X ²¹⁹ can be L X ²²⁰ can be G, X ²²¹ can be Q, X ²²² can be L, X ²²³ can be D, X ²²⁴ can be L and X ²²⁵ can be selected from D and E.

In the IL-2Rβ ligand of formula (11), X ²¹¹ can be selected from H and R, X ²¹² can be W, X ²¹³ can be L, X ²¹⁴ can be P X ²¹⁵ can be W, X ²¹⁶ can be M, X ²¹⁷ can be A, X ²¹⁸ can be Q, X ²¹⁹ can be L X ²²⁰ can be G, X ²²¹ can be Q, X ²²² can be L, X ²²³ can be D, X ²²⁴ can be L and X ²²⁵ can be selected from D and E.

In IL-2Rβ ligands of formula (11), X ²¹¹ can be selected from H and R, X ²¹² can be W, X ²¹³ can be Y and X ²¹⁴ is P X ²¹⁵ can be W, X ²¹⁶ can be M, X ²¹⁷ can be A, X ²¹⁸ can be selected from K and R, X ²¹⁹ can be L, X ²²⁰ can be G, X ²²¹ can be Q, X ²²² can be L, X ²²³ can be D, X ²²⁴ can be L and X ²²⁵ can be selected from D and E.

IL-2Rβ ligands may comprise an amino acid sequence selected from any one of SEQ ID NO:810-SEQ ID NO:903.

The IL-2Rβ ligand can comprise an amino acid sequence selected from any one of SEQ ID NOs:809-903, wherein the amino acid sequence is on the N-terminus, the C-terminus, or both the N-terminus and the C-terminus. It may end with the above amino acids -GG-.

The IL-2Rβ ligand may comprise an amino acid sequence selected from any one of SEQ ID NOs:809-903, each amino acid independently having the following conservative substitutions: alanine (A), glycine (G), amino acids with small hydrophobic side chains including proline (P), serine (S), or threonine (T); hydroxyl-containing sides including serine (S), threonine (T), or tyrosine (Y) amino acids with acidic side chains, including aspartic acid (D), or glutamic acid (E); histidine (H), asparagine (N), glutamine (Q), serine (S), threonine (T), or amino acids with polar neutral side chains, including tyrosine (Y); amino acids with basic side chains, including arginine (R), lysine (K), or histidine (H); isoleucine (I), leucine (L) , methionine (M), valine (V), phenylalanine (F), tyrosine (Y), or tryptophan (W); and phenylalanine (F), histidine (H), tryptophan ( W), or amino acids with aromatic side chains, including tyrosine (Y).

IL-2Rβ ligands of SEQ ID NOs:809-903 can exhibit binding affinities (IC ₅₀ ) for IL-2Rβ subunits of less than 100 μM.

IL-2Rβ ligands can include, for example, 5-50 amino acids, 5-40 amino acids, 5-30 amino acids, 5-30 amino acids, 6-25 amino acids, or 7-20 amino acids.

IL-2Rβ ligand is 1 pM to 100 μM, 10 pM to 10 μM, to human IL-2Rβ subunit, to mammalian IL-2Rβ subunit, or to both human IL-2Rβ subunit and mammalian IL-2Rβ subunit; Binding affinities (IC ₅₀ ) of 100 pM to 1 μM, 0.001 μM to 1 μM, or 0.01 μM to 1 μM can be exhibited.

IL-2Rβ ligands provided by the present disclosure can exhibit binding affinities (IC ₅₀ ) for human IL-2Rβ subunits of, for example, 0.1 μM to 50 μM.

IL-2Rβ ligand is less than 100 μM, less than 10 μM, less than 1 μM to human IL-2Rβ subunit, to mammalian IL-2Rβ subunit, or to both human IL-2Rβ subunit and mammalian IL-2Rβ subunit , less than 0.1 μM, or less than 0.01 μM binding affinities (IC ₅₀ ).

The IL-2Rβ ligand has a binding affinity (IC ₅₀ ) of less than 100 μM, less than 10 μM, less than 1 μM, less than 0.1 μM, or less than 0.01 μM for each of the human IL-2Rβ subunit and the human IL-2Rγc subunit can be shown.

IL-2Rβ ligands have binding affinities of 1 pM to 100 μM, 10 pM to 10 μM, 100 pM to 1 μM, 0.001 μM to 1 μM, or 0.01 μM to 1 μM for human IL-2Rβ subunit and human IL-2Rγc subunit, respectively. properties ( _IC50 ).

IL-2Rβ ligands provided by the present disclosure can exhibit binding affinities (IC ₅₀ ) for human IL-2Rα (CD25) subunits greater than 100 μM, greater than 1 mM, greater than 10 mM, or greater than 100 mM.

The IL-2Rβ ligand is at least 10-fold greater, at least 50-fold greater, at least 100-fold greater, at least 500-fold greater, or at least 1,000-fold greater than the binding affinity of the IL-2Rβ ligand for the human IL-2Rα subunit Binding affinities (IC ₅₀ ) for human IL-2Rβ subunits can be demonstrated.

IL-2Rγc ligands provided by the present disclosure can exhibit a binding affinity (IC ₅₀ ) for the human IL-2Rγc subunit of less than 100 μM, less than 10 μM, less than 1 μM, less than 0.1 μM, or less than 0.01 μM. can.

IL-2Rγc ligands provided by the present disclosure have binding affinities (IC ₅₀ ) can be shown.

IL-2Rγc ligands provided by the present disclosure have binding affinities (IC ₅₀ ) for mammalian IL-2Rγc subunits of, for example, less than 100 μM, less than 10 μM, less than 1 μM, less than 0.1 μM, or less than 0.01 μM. can be shown.

IL-2Rγc ligands provided by the present disclosure have binding affinities for mammalian IL-2Rγc subunits of, for example, 1 pM to 100 μM, 10 pM to 10 μM, 100 pM to 1 μM, 0.001 μM to 1 μM, or 0.01 μM to 1 μM. properties ( _IC50 ).

The IL-2Rγc ligands provided by this disclosure can comprise an amino acid sequence selected from any one of SEQ ID NOs:194-267 and SEQ ID NOs:904-1027.

The IL-2Rγc ligands provided by this disclosure can comprise the amino acid sequence of any one of SEQ ID NOs: 194-267 and SEQ ID NOs: 904-1027, independently of the following conservative Substitutions: amino acids with small hydrophobic side chains, including alanine (A) or glycine (G); amino acids with hydroxyl-containing side chains, including serine (S), threonine (T), or tyrosine (Y); asparagine (I ) or amino acids with acidic and polar neutral side chains, including glutamic acid (E); asparagine (N), or glutamine (Q); phenylalanine (F), tryptophan (W), tyrosine (Y), or histidine (H). amino acids with aromatic side chains, including; amino acids with basic side chains, including arginine (R), lysine (K), or histidine (H); isoleucine (I), leucine (L), methionine (M), Amino acids with large hydrophobic side chains including valine (V), phenylalanine (F), tyrosine (Y), or tryptophan (W), phenylalanine (F), histidine (H), tryptophan (W), or tyrosine (Y) ) containing one or more of the amino acids with aromatic side chains.

The IL-2Rγc ligand can comprise the amino acid sequence of formula (4) (SEQ ID NO: 194) or the amino acid sequence of formula (4a) (SEQ ID NO: 195),
^{-X53 -X54 -X55 -X56 -X57 -X58 -X59 -X60} - (4)
^{-X51 -X52 -CX53 -X54 -X55 -X56 -X57 -X58 -X59 -X60 -CX61 -X62} - (4a)
wherein X ⁵¹ can be selected from G, I, K, L, Q, R, T, Y, and V; X ⁵² can be selected from A, D, E, H, I, L, M, R, S, T, V, and W; X ⁵³ can be selected from D, E, F, N, Q, S, and T; X ⁵⁴ can be selected from A, D, E, G, I, M, N, Q, R, S, and T, and X ⁵⁵ can be selected from D, E, F, Q, S, T, W, and Y X ⁵⁶ can be selected from D, E, F, G, L, M, N, Q, and Y; X ⁵⁷ can be selected from E, G, N, S, and Q; X ⁵⁸ can be selected from I, K, M, P, T, and V; X ⁵⁹ can be selected from I ^, L, M, S, T, and V; can be selected from F, I, and L, X ⁶¹ can be selected from F, T, and W, X ⁶² can be selected from A, E, F, G, I, K, L, M, N, P, Q, T, V, W, and Y can be selected.

In IL-2Rγc ligands of formulas (4) and (4a), ^X51 may be selected from I, L, and V.

In IL-2Rγc ligands of formulas (4) and (4a), ^X52 may be selected from S and T.

In IL-2Rγc ligands of formulas (4) and (4a), ^X53 may be selected from D, E, N, and Q.

In IL-2Rγc ligands of formulas (4) and (4a), ^X54 may be selected from D, E, N, and Q.

In IL-2Rγc ligands of formulas (4) and (4a), ^X55 may be selected from F, W, and Y.

In IL-2Rγc ligands of formulas (4) and (4a), ^X56 may be selected from D, E, N, and Q.

^X57 can be G in the IL-2Rγc ligands of formulas (4) and (4a).

In IL-2Rγc ligands of formulas (4) and (4a), ^X58 may be selected from I and V.

In IL-2Rγc ligands of formulas (4) and (4a), ^X59 may be selected from I, L, M, and V.

In IL-2Rγc ligands of formulas (4) and (4a), ^X60 may be selected from F, I, and L.

In IL-2Rγc ligands of formulas (4) and (4a), ^X61 can be W.

In IL-2Rγc ligands of formulas (4) and (4a), ^X62 may be selected from N and Q.

In IL-2Rγc ligands of formulas (4) and (4a), X ⁵¹ can be selected from I, L, and V, X ⁵² can be selected from S and T, and X ⁵³ can be selected from D, E, N, and Q; X ⁵⁴ can be selected from D ^and N; X ⁵⁵ can be selected from F, W, and Y; D, E, N, and Q, X ⁵⁷ can be G, X ⁵⁸ can be selected from I and V, X ⁵⁹ can be I, L, M, and V, X ⁶⁰ can be selected from F, I, and L, X ⁶¹ can be W, and X ⁶² can be selected from N and Q .

The IL-2Rγc ligand can comprise an amino acid sequence selected from any one of SEQ ID NOs:196-210 and SEQ ID NOs:904-913.

The IL-2Rγc ligand can comprise an amino acid sequence selected from any one of SEQ ID NOs: 194-210 and SEQ ID NOs: 904-913, wherein the amino acid sequence is on the N-terminus, on the C-terminus , or with amino acids -GG- on both the N- and C-termini.

The IL-2Rγc ligand can comprise an amino acid sequence selected from any one of SEQ ID NOs:194-210, and SEQ ID NOs:904-913, each amino acid independently having the following conservation substitution: amino acids with small hydrophobic side chains, including alanine (A), glycine (G), proline (P), serine (S), or threonine (T); serine (S), threonine (T), or Amino acids with hydroxyl-containing side chains, including tyrosine (Y); amino acids with acidic side chains, including aspartic acid (D), or glutamic acid (E); histidine (H), asparagine (N), glutamine (Q), serine. (S), threonine (T), or tyrosine (Y); amino acids with basic side chains, including arginine (R), lysine (K), or histidine (H); amino acids with large hydrophobic side chains including isoleucine (I), leucine (L), methionine (M), valine (V), phenylalanine (F), tyrosine (Y), or tryptophan (W); ), histidine (H), tryptophan (W), or tyrosine (Y).

The IL-2Rγc ligands of SEQ ID NOs:194-210 and SEQ ID NOs:904-913 exhibit binding affinities (IC ₅₀ ) for the IL-2Rγc subunit of less than 100 μM.

The IL-2Rγc ligand can comprise the amino acid sequence of formula (4) (SEQ ID NO: 194) or the amino acid sequence of formula (4a) (SEQ ID NO: 195),
^{-X53 -X54 -X55 -X56 -X57 -X58 -X59 -X60} - (4)
^{-X51 -X52 -CX53 -X54 -X55 -X56 -X57 -X58 -X59 -X60 -CX61 -X62} - (4a)
wherein X ⁵¹ can be selected from amino acids, X ⁵² can be selected from amino acids, X ⁵³ can be selected from amino acids containing polar neutral side chains or acidic side chains, X ⁵⁴ can be selected from amino acids comprising a polar neutral side chain or an acidic side chain, X ⁵⁵ can be selected from amino acids, X ⁵⁶ can be selected from amino acids, X ⁵⁷ can be selected from , can be selected from amino acids containing small hydrophobic side chains, X ⁵⁸ can be selected from amino acids containing large hydrophobic side chains, and X ⁵⁹ can be selected from amino acids containing large hydrophobic side chains. X ⁶⁰ can be selected from amino acids with large hydrophobic side chains, X ⁶¹ can be selected from amino acids with large hydrophobic side chains, and X ⁶² is selected from amino acids be able to.

In the IL-2Rγc ligands of formulas (4) and (4a), ^X51 can be selected from amino acids containing large hydrophobic and basic side chains, and ^X52 can be selected from hydroxyl-containing side chains and large hydrophobic side chains. ^X53 may be selected from amino acids containing a polar neutral side chain or an acidic side chain; ^X54 may be selected from amino acids containing a polar neutral side chain or an acidic side chain; ^X55 can be selected from amino acids containing a large hydrophobic side chain, ^X56 can be selected from amino acids containing a polar neutral side chain or an acidic side chain, ^X57 can be selected from amino acids containing a small hydrophobic side chain X ⁵⁸ can be selected from amino acids containing large hydrophobic side chains, X ⁵⁹ can be selected from amino acids containing large hydrophobic side chains, X ⁶⁰ can be selected from amino acids containing large hydrophobic side chains, X ⁶¹ can be selected from amino acids containing large hydrophobic side chains, and X ⁶² can be selected from amino acids containing polar neutral side chains. can be selected.

In IL-2Rγc ligands of formulas (4) and (4a), X ⁵¹ can be selected from R, K, H, F, I, L, M, V, Y, and W, and X ⁵² is S, T, F, I, L, M, V, Y, and W, and X ⁵³ can be selected from D, E, H, N, Q, S, T, and Y X ⁵⁴ can be selected from D, E, H, N, Q, S, T, and Y, and X ⁵⁵ can be selected from F, I, L, M, V, Y, and W X ⁵⁶ can be selected from D, E, H, N, Q, S, T, and Y, and X ⁵⁷ can be selected from A, G, P, S, and T X ⁵⁸ can be selected from F, I, L, M, V, Y, and W; X ⁵⁹ can be selected from F, I, L, M, V, Y, and W; X ⁶⁰ can be selected from F, I, L, M, V, Y, and W; X ⁶¹ can be selected from F, I, L, M, V, Y, and W; and X ⁶² can be selected from H, N, Q, S, T, and Y.

In IL-2Rγc ligands of formulas (4) and (4a), ^X51 may be selected from I, L, and V.

In IL-2Rγc ligands of formulas (4) and (4a), ^X52 may be selected from S and T.

In IL-2Rγc ligands of formulas (4) and (4a), ^X53 may be selected from D, E, and Q.

In IL-2Rγc ligands of formulas (4) and (4a), ^X54 may be selected from D, E, and N.

In IL-2Rγc ligands of formulas (4) and (4a), ^X55 may be selected from F, Y, and W.

In IL-2Rγc ligands of formulas (4) and (4a), ^X56 may be selected from D, E, N, and Q.

^X57 can be G in the IL-2Rγc ligands of formulas (4) and (4a).

In IL-2Rγc ligands of formulas (4) and (4a), ^X58 may be selected from I and V.

In IL-2Rγc ligands of formulas (4) and (4a), ^X59 may be selected from I, L, M, and V.

In IL-2Rγc ligands of formulas (4) and (4a), ^X60 may be selected from F, I, and L.

In IL-2Rγc ligands of formulas (4) and (4a), ^X61 can be W.

In IL-2Rγc ligands of formulas (4) and (4a), ^X62 may be selected from N and Q.

In IL-2Rγc ligands of formulas (4) and (4a), X ⁵¹ can be selected from I, L, and V, X ⁵² can be selected from S and T, and X ⁵³ can be selected from D, E, and Q, X ⁵⁴ can be selected from D, E, and N, X ⁵⁵ can be selected from F, Y, and W, and X ⁵⁶ can be selected from , D, E, N, and Q, X ⁵⁷ can be G, X ⁵⁸ can be selected from I and V, X ⁵⁹ can be I, L, M , and V, X ⁶⁰ may be selected from F, I, and L, X ⁶¹ may be W, and X ⁶² may be selected from N and Q can be done.

In the IL-2Rγc ligands of formulas (4) and (4a), X ⁵¹ can be selected from G, I, K, L, Q, R, and V, and X ⁵² can be A, D, E, H, I, L, M, R, S, T, V, and W; X ⁵³ can be selected from D, E, F, N, Q, S, and T; X ⁵⁴ can be selected from A, D, E, G, I, M, N, R, S, and T, and X ⁵⁵ can be selected from D, E, F, Q, S, T, W, and Y, X ⁵⁶ can be selected from D, E, F, G, L, M, N, Q, S, and Y; X ⁵⁷ can be selected from C, E, G, N , Q, and S, X ⁵⁸ can be selected from I, P, T, and V, and X ⁵⁹ can be selected from I, K, L, M, P, S, T, and V can be selected, X ⁶⁰ can be selected from F, I, and L, X ⁶¹ can be selected from F, T, and W, X ⁶² can be selected from A, E, F, G, I, K, M, N, Q, T, and W can be selected.

In IL-2Rγc ligands of formulas (4) and (4a), X ⁵¹ can be selected from I, L, and V, X ⁵² can be selected from S and T, and X ⁵³ can be selected from D, E, N, and Q; X ⁵⁴ can be selected from D, E, N, S, and T; X ⁵⁵ can be selected from F, S, T, W, and Y X ⁵⁶ can be selected from D, E, N, and Q; X ⁵⁷ can be selected from G and N; X ⁵⁸ can be selected from I and V X ⁵⁹ can be selected from I, L, M, and V; X ⁶⁰ can be selected from F, I, and L; X ⁶¹ is ^W ; can be selected from N and Q.

In IL-2Rγc ligands of formulas (4) and (4a), X ⁵¹ can be selected from I, L, and V, X ⁵² can be selected from S and T, and X ⁵³ can be selected from Q, X ⁵⁴ can be selected from D, E, N, S, and T, X ⁵⁵ can be selected from S, T, and W, X ⁵⁶ can be selected from D , E, N, and Q, X ⁵⁷ can be G, X ⁵⁸ can be V, X ⁵⁹ can be L, and X ⁶⁰ can be , L, X ⁶¹ can be W, and X ⁶² can be selected from N and Q.

The IL-2Rγc ligand may comprise the amino acid sequence of formula (5) (SEQ ID NO:211) or formula (5a) (SEQ ID NO:212),
^{-X73 -X74 -X75 -X76 -X77 -X78 -X79 -X80} - (5)
^{-X71 -X72 -CX73 -X74 -X75 -X76 -X77 -X78 -X79 -X80 -CX81 -X82} - (5a)
wherein X ⁷¹ can be selected from F, G, I, L, P, Q, R, T, and V; X ⁷² can be selected from A, D, E, I, M, R, S, T, and V; X ⁷³ can be selected from D, E, F, M, N, Q, S, T, V, W, and Y; X ⁷⁴ can be selected from D, E, F, G, I, L, M, P, R, S, T, and V; X ⁷⁵ can be selected from F, H, L, W, and Y; X ⁷⁶ can be selected from D, E, H, L, N, Q, S, and T; ^{X 77} can be selected from G, T, Q, and E; I, L, M, Q, and V, X ⁷⁹ can be selected from D, E, N, Q, and R, X ⁸⁰ can be selected from D, F, I, and L X ⁸¹ can be selected from F, I, L, R, T, W, and Y; X ⁸² can be selected from A, F, G, H, I, N, P, Q , S, T, and W.

In IL-2Rγc ligands of formulas (5) and (5a), ^X71 may be selected from I, L, and V.

In IL-2Rγc ligands of formulas (5) and (5a), ^X72 can be selected from A, D, E, I, M, and V.

In IL-2Rγc ligands of formulas (5) and (5a), ^X73 may be selected from E, Q, and N.

In IL-2Rγc ligands of formulas (5) and (5a), ^X74 may be selected from D and E.

In IL-2Rγc ligands of formulas (5) and (5a), ^X75 can be selected from F, W, and Y.

In IL-2Rγc ligands of formulas (5) and (5a), ^X76 may be selected from D, E, L, N, and Q.

^X77 can be G in IL-2Rγc ligands of formulas (5) and (5a).

In IL-2Rγc ligands of formulas (5) and (5a), ^X78 may be selected from I, M, and V.

In IL-2Rγc ligands of formulas (5) and (5a), ^X79 may be selected from D, E, Q, and R.

In IL-2Rγc ligands of formulas (5) and (5a), X ⁸⁰ may be selected from F, I, and L.

In IL-2Rγc ligands of formulas (5) and (5a), ^X81 can be W.

In IL-2Rγc ligands of formulas (5) and (5a), ^X82 may be selected from N and Q.

In IL-2Rγc ligands of formulas (5) and (5a), X ⁷¹ can be selected from I, L, and V, and X ⁷² is selected from A, D, E, I, M, and V. X ⁷³ can be selected from E, Q, and N; X ⁷⁴ can be selected from D and E; and X ⁷⁵ can be selected from F, W, and Y , X ⁷⁶ can be selected from D, E, L, N, and Q, X ⁷⁷ can be G, and X ⁷⁸ can be selected from I, M, and V X ⁷⁹ can be selected from D, E, Q, and R; X ⁸⁰ can be selected from F, I, and L; X ⁸¹ can be W; ⁸² can be selected from N and Q;

The IL-2Rγc ligand may comprise the amino acid sequence of formula (5) (SEQ ID NO:211) or formula (5a) (SEQ ID NO:212),
^{-X73 -X74 -X75 -X76 -X77 -X78 -X79 -X80} - (5)
^{-X71 -X72 -CX73 -X74 -X75 -X76 -X77 -X78 -X79 -X80 -CX81 -X82} - (5a)
wherein X ⁶¹ can be selected from amino acids, X ⁶² can be selected from amino acids, X ⁶³ can be selected from amino acids, X ⁶⁴ can be selected from amino acids , X ⁶⁵ can be selected from amino acids containing large hydrophobic side chains, X ⁶⁶ can be selected from amino acids, X ⁶⁷ can be selected from small hydrophobic side chains, X ⁶⁸ can be selected from amino acids containing large hydrophobic side chains, X ⁶⁹ can be selected from amino acids containing basic, acidic, or polar neutral side chains, and X ⁷⁰ can be selected from It can be selected from amino acids containing large hydrophobic side chains, ^X71 can be selected from amino acids containing large hydrophobic side chains, and ^X72 can be selected from amino acids.

In the IL-2Rγc ligands of formulas (5) and (5a), ^X71 can be selected from amino acids containing large hydrophobic side chains and ^X72 can be selected from amino acids containing acidic or large hydrophobic side chains. X ⁷³ can be selected from an amino acid containing an acidic side chain, a hydroxyl containing side chain, or a polar neutral side chain, X ⁷⁴ can be selected from an acidic side chain, a hydroxyl containing side chain, or can be selected from amino acids containing large hydrophobic side chains, X ⁷⁵ can be selected from amino acids containing large hydrophobic side chains, X ⁷⁶ can be selected from acidic side chains, hydroxyl-containing side chains, or X ⁷⁷ can be selected from amino acids containing small hydrophobic side chains, X ⁷⁸ can be selected from amino acids containing large hydrophobic side chains, X ⁷⁹ can be selected from amino acids containing basic, acidic or polar neutral side chains, ^X80 can be selected from amino acids containing large hydrophobic side chains, and ^X81 can be selected from It can be selected from amino acids containing large hydrophobic side chains and ^X82 can be selected from amino acids containing polar neutral side chains.

In the IL-2Rγc ligands of formulas (5) and (5a), X ⁷¹ can be selected from F, I, L, M, V, Y, and W, and X ⁷² can be D, E, F, I, L, M, V, Y, and W; X ⁷³ may be selected from D, E, S, T, H, N, Q, S, T, and Y; X ⁷⁴ can be selected from D, E, S, T, F, I, L, V, Y, and W; X ⁷⁵ can be selected from F, I, L, M, V, Y, and W; X ⁷⁶ can be selected from D, E, S, T, H, N, Q, S, T, and Y; X ⁷⁷ can be selected from A, G, P, S, and T, X ⁷⁸ can be selected from F, I, L, M, V, Y, and W; X ⁷⁹ can be selected from R, K, H, D, E, H, N , Q, S, T, and Y; X ⁸⁰ may be selected from F, I, L, M, V, Y, and W; X ⁸¹ may be selected from F, I, L , M, V, Y, and W, and X ⁸² may be selected from H, N, Q, S, T, and Y.

In IL-2Rγc ligands of formulas (5) and (5a), ^X71 may be selected from I, L, and V.

In IL-2Rγc ligands of formulas (5) and (5a), ^X72 can be selected from D, E, I, M, and V.

In IL-2Rγc ligands of formulas (5) and (5a), ^X73 may be selected from E, N, and Q.

In IL-2Rγc ligands of formulas (5) and (5a), ^X74 may be selected from D and E.

In IL-2Rγc ligands of formulas (5) and (5a), ^X75 can be selected from F, W, and Y.

In IL-2Rγc ligands of formulas (5) and (5a), ^X76 may be selected from D, E, and N.

^X77 can be G in IL-2Rγc ligands of formulas (5) and (5a).

In IL-2Rγc ligands of formulas (5) and (5a), ^X78 may be selected from I, M, and V.

In IL-2Rγc ligands of formulas (5) and (5a), ^X79 may be selected from D, E, N, Q, and R.

In IL-2Rγc ligands of formulas (5) and (5a), X ⁸⁰ may be selected from F, I, and L.

In IL-2Rγc ligands of formulas (5) and (5a), ^X81 can be W.

In IL-2Rγc ligands of formulas (5) and (5a), ^X82 may be selected from N and Q.

In the IL-2Rγc ligands of formulas (5) and (5a), ^X71 can be selected from I, L, and V, and ^X72 can be selected from D, E, I, M, and V. , X ⁷³ can be selected from E, N, and Q, X ⁷⁴ can be selected from D and E, and X ⁷⁵ can be selected from F, W, and Y , X ⁷⁶ can be selected from D, E, and N, X ⁷⁷ can be G, X ⁷⁸ can be selected from I, M, and V, and X ⁷⁹ can be selected from D, E, N, Q, and R; X ⁸⁰ can be selected from F ^, I, and L; X ⁸¹ can be W; and Q.

In the IL-2Rγc ligands of formulas (5) and (5a), ^{X 71} can be selected from F, G, I, L, P, Q, R, T, and V; D, E, I, M, L, M, R, S, T, and V, where X ⁷³ is D, E, F, M, N, Q, S, T, V, W , and Y, X ⁷⁴ can be selected from D, E, F, G, I, L, M, P, R, S, T, and V, and X ⁷⁵ can be selected from F , H, L, W, and Y, X ⁷⁶ can be selected from D, E, H, L, N, Q, S, and T, and X ⁷⁷ can be selected from E, G , Q, and T, X ⁷⁸ can be selected from I, L, M, Q, and V, and X ⁷⁹ can be selected from D, E, N, Q, and R X ⁸⁰ can be selected from D, F, I, and L, and X ⁸¹ can be selected from C, F, I, L, Q, R, T, W, and Y and X ⁸² can be selected from A, F, G, H, I, L, N, P, Q, T, and W.

In IL-2Rγc ligands of formulas (5) and (5a), X ⁷¹ can be selected from F, I, L, and V; X ⁷² can be D, E, I, S, T, and V; X ⁷³ can be selected from D, E, N, and Q; X ⁷⁴ can be selected from D, E, F, I, L, M, and V , X ⁷⁵ can be selected from F, W, and Y; X ⁷⁶ can be selected from D, E, N, and Q; X ⁷⁷ can be ^G ; can be selected from I, L, M, and V, X ⁷⁹ can be selected from D, E, N, Q, and R, and X ⁸⁰ can be selected from D, F, I, and L and X ⁸¹ may be selected from F, I, L, and W, and X ⁸² may be selected from F, I, L, N, Q, and W.

In IL-2Rγc ligands of formulas (5) and (5a), X ⁷¹ can be selected from F, I, L, and V; X ⁷² can be D, E, I, S, T, and V; X ⁷³ can be selected from D, E, N, and Q; X ⁷⁴ can be selected from D, E, F, I, L, M, and V , X ⁷⁵ can be W, X ⁷⁶ can be selected from D, E, N, and Q, X ⁷⁷ can be G, and X ⁷⁸ can be V , X ⁷⁹ can be selected from D, E, N, Q, and R, X ⁸⁰ can be L, X ⁸¹ can be W, and X ⁸² can be , F, I, L, N, Q, and W.

The IL-2Rγc ligand can comprise an amino acid sequence selected from any one of SEQ ID NOs:213-233 and SEQ ID NOs:914-920.

The IL-2Rγc ligand can comprise an amino acid sequence selected from any one of SEQ ID NOs:211-233 and SEQ ID NOs:914-920, wherein the amino acid sequence is on the N-terminus, on the C-terminus , or with amino acids -GG- on both the N- and C-termini.

The IL-2Rγc ligand can comprise an amino acid sequence selected from any one of SEQ ID NOs:211-233, and SEQ ID NOs:914-920, each amino acid independently having the following conservation substitution: amino acids with small hydrophobic side chains, including alanine (A), glycine (G), proline (P), serine (S), or threonine (T); serine (S), threonine (T), or Amino acids with hydroxyl-containing side chains, including tyrosine (Y); amino acids with acidic side chains, including aspartic acid (D), or glutamic acid (E); histidine (H), asparagine (N), glutamine (Q), serine. (S), threonine (T), or tyrosine (Y); amino acids with basic side chains, including arginine (R), lysine (K), or histidine (H); amino acids with large hydrophobic side chains including isoleucine (I), leucine (L), methionine (M), valine (V), phenylalanine (F), tyrosine (Y), or tryptophan (W); ), histidine (H), tryptophan (W), or tyrosine (Y).

The IL-2Rγc ligands of SEQ ID NOs:211-233 and SEQ ID NOs:914-920 exhibit binding affinities (IC ₅₀ ) for the IL-2Rγc subunit of less than 100 μM.

The IL-2Rγc ligand may comprise the amino acid sequence of formula (6) (SEQ ID NO:234) or formula (6a) (SEQ ID NO:235),
^{-X93 -X94 -X95 -X96 -X97 -X98 -X99 -X100 -X101} - (6)
^{-X91 -X92 -X93 -X94 -X95 -X96 -X97 -X98 -X99 -X100 -X101 -X102 -X103} - (6a)
wherein X ⁹¹ can be selected from C, D, E, and L; X ⁹² can be selected from C, L, M, R, S, V, and ^W ; , C, D, F, P, and R, X ⁹⁴ can be selected from A, D, L, Q, S, and W, and X ⁹⁵ can be selected from D, E, F , L, and V, X ⁹⁶ can be selected from A, D, E, F, G, K, Q, and S, and X ⁹⁷ can be selected from E, L, M, and W, X ⁹⁸ can be selected from G, I, L, W, and Y, X ⁹⁹ can be selected from E, I, R, T, and V; X ¹⁰⁰ can be W, X ¹⁰¹ can be selected from C, A, I, L, P, and V, and X ¹⁰² can be selected from C, D, G, H and X ¹⁰³ can be selected from C, D, E, H, S, and T.

In IL-2Rγc ligands of formulas (6) and (6a), ^X91 may be selected from D and E.

In IL-2Rγc ligands of formulas (6) and (6a), ^X92 can be selected from L, M, R, S, V, and W.

In IL-2Rγc ligands of formulas (6) and (6a), ^X93 may be selected from D and F.

^X94 can be S in IL-2Rγc ligands of formulas (6) and (6a).

In IL-2Rγc ligands of formulas (6) and (6a), ^X95 may be selected from D and E.

In IL-2Rγc ligands of formulas (6) and (6a), ^X96 may be selected from D and E.

In IL-2Rγc ligands of formulas (6) and (6a), ^X97 can be selected from L, M, and W.

X ⁹⁸ can be G in IL-2Rγc ligands of formulas (6) and (6a).

X ⁹⁹ can be E in IL-2Rγc ligands of formulas (6) and (6a).

X ¹⁰⁰ can be W in IL-2Rγc ligands of formulas (6) and (6a).

In IL-2Rγc ligands of formulas (6) and (6a), X ¹⁰¹ may be selected from I, L, and V.

In IL-2Rγc ligands of formulas (6) and (6a), X ¹⁰² may be selected from D and G.

In IL-2Rγc ligands of formulas (6) and (6a), X ¹⁰³ may be selected from S and T.

In IL-2Rγc ligands of formulas (6) and (6a), X ⁹¹ can be selected from D and E and X ⁹² can be selected from L, M, R, S, V, and W. X ⁹³ can be selected from D and F, X ⁹⁴ can be S, X ⁹⁵ can be selected from D and E, X ⁹⁶ can be selected from D and E X ⁹⁷ can be selected from L, M, and W; ^{X 98} can be G; X ⁹⁹ can be E; and X ¹⁰¹ can be selected from I, L, and V, X ¹⁰² can be selected from D and G, and X ¹⁰³ can be selected from S and T.

IL-2Rγc ligands comprise an amino acid sequence selected from any one of SEQ ID NOs:236-245.

The IL-2Rγc ligand can comprise an amino acid sequence selected from any one of SEQ ID NOS:234-245, wherein the amino acid sequence is on the N-terminus, the C-terminus, or both the N-terminus and the C-terminus. It may end with the above amino acids -GG-.

The IL-2Rγc ligand may comprise an amino acid sequence selected from any one of SEQ ID NOS:234-245, each amino acid independently having the following conservative substitutions: alanine (A), glycine (G), amino acids with small hydrophobic side chains including proline (P), serine (S), or threonine (T); hydroxyl-containing sides including serine (S), threonine (T), or tyrosine (Y) amino acids with acidic side chains, including aspartic acid (D), or glutamic acid (E); histidine (H), asparagine (N), glutamine (Q), serine (S), threonine (T), or amino acids with polar neutral side chains, including tyrosine (Y); amino acids with basic side chains, including arginine (R), lysine (K), or histidine (H); isoleucine (I), leucine (L) , methionine (M), valine (V), phenylalanine (F), tyrosine (Y), or tryptophan (W); and phenylalanine (F), histidine (H), tryptophan ( W), or amino acids with aromatic side chains, including tyrosine (Y).

The IL-2Rγc ligands of SEQ ID NOs:234-245 exhibit binding affinities (IC ₅₀ ) for the IL-2Rγc subunit of less than 100 μM.

The IL-2Rγc ligand may comprise the amino acid sequence of formula (6) (SEQ ID NO:234) or formula (6a) (SEQ ID NO:235),
^{-X93 -X94 -X95 -X96 -X97 -X98 -X99 -X100 -X101} - (6)
^{-X91 -X92 -X93 -X94 -X95 -X96 -X97 -X98 -X99 -X100 -X101 -X102 -X103} - (6a)
wherein X ⁹¹ can be selected from amino acids or cysteine with an acidic side chain, X ⁹² can be selected from amino acids, X ⁹³ is an amino acid with an acidic side chain or a large hydrophobic side chain X ⁹⁴ can be selected from amino acids, X ⁹⁵ can be selected from amino acids, X ⁹⁶ can be selected from amino acids, X ⁹⁷ can be selected from a large hydrophobic can be selected from amino acids containing side chains, X ⁹⁸ can be selected from amino acids containing small hydrophobic side chains or large hydrophobic side chains, X ⁹⁹ can be selected from amino acids, X ¹⁰⁰ can be selected from amino acids containing large hydrophobic side chains, X ¹⁰¹ can be selected from amino acids containing large hydrophobic side chains, and X ¹⁰² can be selected from small hydrophobic or acidic side chains. or cysteine containing amino acids and X ¹⁰³ can be selected from amino acids containing acidic side chains or hydroxyl containing side chains or cysteine.

In the IL-2Rγc ligands of formulas (6) and (6a), X ⁹¹ can be selected from amino acids containing acidic side chains, X ⁹² can be selected from amino acids, and X ⁹³ can be selected from acidic side chains. or a large hydrophobic side chain, ^X94 can be selected from amino acids with an acidic side chain or a hydroxyl-containing side chain, ^X95 can be selected from an amino acid with an acidic side chain. can be selected, X ⁹⁶ can be selected from amino acids, X ⁹⁷ can be selected from amino acids containing large hydrophobic side chains, X ⁹⁸ can be selected from small hydrophobic side chains or large hydrophobic can be selected from amino acids containing side chains, X ⁹⁹ can be selected from amino acids containing acidic or large hydrophobic side chains, and X ¹⁰⁰ can be selected from amino acids containing large hydrophobic side chains X ¹⁰¹ can be selected from amino acids containing large hydrophobic side chains, X ¹⁰² can be selected from amino acids containing small hydrophobic or acidic side chains, and X ¹⁰³ can be selected from It can be selected from amino acids containing acidic side chains or hydroxyl containing side chains.

In the IL-2Rγc ligands of formulas (6) and (6a), X ⁹¹ can be selected from D and E, X ⁹² can be selected from amino acids, X ⁹³ can be D, E, F , I, L, M, V, Y, and W; X ⁹⁴ may be selected from D, E, S, and T; X ⁹⁵ may be selected from D and E; X ⁹⁶ can be selected from amino acids, X ⁹⁷ can be selected from F, I, L, M, V, Y, and W, X ⁹⁸ can be selected from A, G, P, S, T, F, I, L, M, V, Y, and W, and X ⁹⁹ is selected from D, E, F, I, L, M, V, Y, and W X ¹⁰⁰ can be selected from F, I, L, M, V, Y, and W, and X ¹⁰¹ can be selected from F, I, L, M, V, Y, and W , X ¹⁰² can be selected from D, E, A, G, P, S, and T, and X ¹⁰³ can be selected from D, E, S, and T.

In the IL-2Rγc ligands of formulas (6)-(6a), ^X91 may be selected from D and E.

In IL-2Rγc ligands of formulas (6)-(6a), ^X92 may be selected from amino acids.

In IL-2Rγc ligands of formulas (6) and (6a), ^X93 may be selected from D and F.

^X94 can be S in IL-2Rγc ligands of formulas (6) and (6a).

In IL-2Rγc ligands of formulas (6) and (6a), ^X95 may be selected from D and E.

In IL-2Rγc ligands of formulas (6) and (6a), ^X96 may be selected from amino acids.

In IL-2Rγc ligands of formulas (6) and (6a), ^X97 can be selected from L, M, and W.

X ⁹⁸ can be G in IL-2Rγc ligands of formulas (6) and (6a).

X ⁹⁹ can be E in IL-2Rγc ligands of formulas (6) and (6a).

X ¹⁰⁰ can be W in IL-2Rγc ligands of formulas (6) and (6a).

In IL-2Rγc ligands of formulas (6) and (6a), X ¹⁰¹ may be selected from I, L, and V.

In IL-2Rγc ligands of formulas (6) and (6a), X ¹⁰² may be selected from D and G.

In IL-2Rγc ligands of formulas (6) and (6a), X ¹⁰³ may be selected from S and T.

In the IL-2Rγc ligands of formulas (6) and (6a), X ⁹¹ can be selected from D and E, X ⁹² can be selected from amino acids, X ⁹³ can be selected from D and F X ⁹⁴ can be S, X ⁹⁵ can be selected from D and E, X ⁹⁶ can be selected from amino acids, X ⁹⁷ can be L, M, and W, X ⁹⁸ can be G, X ⁹⁹ can be E, X ¹⁰⁰ can be W, X ¹⁰¹ can be I, L, and V, X ¹⁰² can be selected from D and G, and X ¹⁰³ can be selected from S and T.

The IL-2Rγc ligand may comprise the amino acid sequence of formula (7) (SEQ ID NO:246) or formula (7a) (SEQ ID NO:247),
^{-X114 -X115 -CX116 -X117 -X118} - (7)
^{-X111 -X112 -X113 -X114 -X115 -CX116 -X117 -X108 -X119 -X120 -X121} - (7a)
wherein X ¹¹¹ can be selected from D, G, I, and Q, X ¹¹² can be selected from D, I, and L, X ¹¹³ can be selected from G, L, M, Q , R, S, and Y, X ¹¹⁴ can be selected from D, E, G, L, S, T, and Y, and X ¹¹⁵ can be selected from E, L, P, and Q, X ¹¹⁶ can be selected from D, E, K, L, S, and T, X ¹¹⁷ can be selected from D, F, S, and W; X ¹¹⁸ can be selected from F, N, W, and Y; X ¹¹⁹ can be selected from F, I, L, R, and W; X ¹²⁰ can be selected from A, E, L, and S, and X ¹²¹ can be selected from H, I, K, N, Q, and V.

In IL-2Rγc ligands of formulas (7) and (7a), X ¹¹¹ may be selected from D and Q.

In IL-2Rγc ligands of formulas (7) and (7a), X ¹¹² may be selected from I and L.

In IL-2Rγc ligands of formulas (7) and (7a), X ¹¹³ may be selected from G, L, M, R, S, and Y.

X ¹¹⁴ can be L in IL-2Rγc ligands of formulas (7) and (7a).

In IL-2Rγc ligands of formulas (7) and (7a), X ¹⁵ may be selected from E and Q.

In IL-2Rγc ligands of formulas (7) and (7a), X ¹¹⁶ may be selected from D and E.

In IL-2Rγc ligands of formulas (7) and (7a), X ¹¹⁷ may be selected from F and W.

In IL-2Rγc ligands of formulas (7) and (7a), X ¹¹⁸ can be selected from F, W, and Y.

In IL-2Rγc ligands of formulas (7) and (7a), X ¹¹⁹ may be selected from F, I, and L.

X ¹²⁰ can be S in IL-2Rγc ligands of formulas (7) and (7a).

In IL-2Rγc ligands of formulas (7) and (7a), X ¹²¹ may be selected from N and Q.

In the IL-2Rγc ligands of formulas (7) and (7a), X ¹¹¹ can be selected from D and Q, X ¹¹² can be selected from I and L, X ¹¹³ can be G, L , M, R, S, and Y, X ¹¹⁴ can be L, X ¹¹⁵ can be selected from E and Q, and X ¹¹⁶ can be selected from D and E , X ¹¹⁷ can be selected from F and W, X ¹¹⁸ can be selected from F, W, and Y, and X ¹¹⁹ can be selected from F, I, and L , X ¹²⁰ can be S, and X ¹²¹ can be selected from N and Q.

In the IL-2Rγc ligands of formulas (7) and (7a), X ¹¹¹ can be selected from D, G, I, Q, and W, and X ¹¹² is selected from C, D, I, and L. X ¹¹³ can be selected from G, L, M, Q, R, S, and Y, and X ¹¹⁴ can be selected from D, E, G, L, Q, S, T, and Y , X ¹¹⁵ can be selected from E, G, L, P, and Q, and X ¹¹⁶ can be selected from D, E, K, L, S, and T , X ¹¹⁷ can be selected from D, F, S, and W, X ¹¹⁸ can be selected from F, N, W, and Y, and X ¹¹⁹ can be selected from F, I, L, R , and W, X ¹²⁰ can be selected from A, C, E, L, and S, and X ¹²¹ can be selected from H, I, K, N, Q, and V be able to.

In the IL-2Rγc ligands of formulas (7) and (7a), X ¹¹¹ can be selected from D and Q, X ¹¹² can be selected from I and L, X ¹¹³ can be G, L , M, Q, R, S, and Y, X ¹¹⁴ can be selected from D and S, X ¹¹⁵ can be L, X ¹¹⁶ can be D and E X ¹¹⁷ can be selected from F and W; X ¹¹⁸ can be selected from F, W, and Y; X ¹¹⁹ can be selected from F, I, L, and W X ¹²⁰ can be selected from L and S, and X ¹²¹ can be selected from N and Q.

The IL-2Rγc ligand can comprise an amino acid sequence selected from any one of SEQ ID NOs:248-254 and SEQ ID NOs:921-922.

The IL-2Rγc ligand can comprise an amino acid sequence selected from any one of SEQ ID NOs:246-254 and SEQ ID NOs:921-922, wherein the amino acid sequence is on the N-terminus, on the C-terminus , or with amino acids -GG- on both the N- and C-termini.

The IL-2Rγc ligand can comprise an amino acid sequence selected from any one of SEQ ID NOs:246-254, and SEQ ID NOs:921-922, each amino acid independently having the following conservation substitution: amino acids with small hydrophobic side chains, including alanine (A), glycine (G), proline (P), serine (S), or threonine (T); serine (S), threonine (T), or Amino acids with hydroxyl-containing side chains, including tyrosine (Y); amino acids with acidic side chains, including aspartic acid (D), or glutamic acid (E); histidine (H), asparagine (N), glutamine (Q), serine. (S), threonine (T), or tyrosine (Y); amino acids with basic side chains, including arginine (R), lysine (K), or histidine (H); amino acids with large hydrophobic side chains including isoleucine (I), leucine (L), methionine (M), valine (V), phenylalanine (F), tyrosine (Y), or tryptophan (W); ), histidine (H), tryptophan (W), or tyrosine (Y).

The IL-2Rγc ligands of SEQ ID NOs:246-254, SEQ ID NOs:921-922 exhibit binding affinities (IC ₅₀ ) for the IL-2Rγc subunit of less than 100 μM.

The IL-2Rγc ligand may comprise the amino acid sequence of formula (7) (SEQ ID NO:246) or formula (7a) (SEQ ID NO:247),
^{-X114 -X115 -CX116 -X117 -X118} - (7)
^{-X111 -X112 -X113 -X114 -X115 -CX116 -X117 -X118 -X119 -X120 -X121} - (7a)
wherein X ¹¹¹ can be selected from amino acids, X ¹¹² can be selected from amino acids containing large hydrophobic or acidic side chains, X ¹¹³ can be selected from amino acids, X ¹¹⁴ can be selected from amino acids containing acidic or hydroxyl-containing side chains, X ¹¹⁵ can be selected from amino acids, X ¹¹⁶ can be selected from amino acids, X ¹¹⁷ can be selected from can be selected from amino acids with large hydrophobic side chains, X ¹¹⁸ can be selected from amino acids with large hydrophobic side chains, and X ¹¹⁹ can be selected from amino acids with large hydrophobic side chains; X ¹²⁰ can be selected from amino acids and X ¹²¹ can be selected from amino acids.

In the IL-2Rγc ligands of formulas (7) and (7a), X ¹¹¹ can be selected from amino acids, X ¹¹² can be selected from amino acids containing large hydrophobic or acidic side chains, X ¹¹³ can be selected from amino acids, X ¹¹⁴ can be selected from amino acids containing acidic or hydroxyl-containing side chains, and X ¹¹⁵ can be selected from amino acids containing large hydrophobic side chains. X ¹¹⁶ can be selected from amino acids containing acidic side chains, X ¹¹⁷ can be selected from amino acids containing large hydrophobic side chains, and X ¹¹⁸ can be selected from amino acids containing large hydrophobic side chains. can be selected from amino acids, X ¹¹⁹ can be selected from amino acids containing large hydrophobic side chains, X ¹²⁰ can be selected from amino acids, and X ¹²¹ can be selected from amino acids containing polar neutral side chains It can be selected from amino acids.

In the IL-2Rγc ligands of formulas (7) and (7a), X ¹¹¹ can be selected from amino acids and X ¹¹² can be selected from D, E, F, I, L, M, V, Y, and W. X ¹¹³ can be selected from amino acids, X ¹¹⁴ can be selected from D, E, S, and T, X ¹¹⁵ can be selected from F, I, L, M, V , Y, and W, X ¹¹⁶ can be selected from D and E, and X ¹¹⁷ can be selected from F, I, L, M, V, Y, and W , X ¹¹⁸ can be selected from F, I, L, M, V, Y, and W, and X ¹¹⁹ can be selected from F, I, L, M, V, Y, and W. , X ¹²⁰ can be selected from amino acids and X ¹²¹ can be selected from H, N, Q, S, T, and Y.

In IL-2Rγc ligands of formulas (7) and (7a), X ¹¹¹ may be selected from amino acids.

In IL-2Rγc ligands of formulas (7) and (7a), X ¹¹² may be selected from I and L.

In IL-2Rγc ligands of formulas (7) and (7a), X ¹¹³ may be selected from amino acids.

In IL-2Rγc ligands of formulas (7) and (7a), X ¹¹⁴ may be selected from D, E, and S.

X ¹¹⁵ can be L in IL-2Rγc ligands of formulas (7) and (7a).

In IL-2Rγc ligands of formulas (7) and (7a), X ¹¹⁶ may be selected from D and E.

In IL-2Rγc ligands of formulas (7) and (7a), X ¹¹⁷ may be selected from F and W.

In IL-2Rγc ligands of formulas (7) and (7a), X ¹¹⁸ can be selected from F, W, and Y.

In IL-2Rγc ligands of formulas (7) and (7a), X ¹¹⁹ may be selected from F, I, and L.

In IL-2Rγc ligands of formulas (7) and (7a), X ¹²⁰ may be selected from amino acids.

In IL-2Rγc ligands of formulas (7) and (7a), X ¹²¹ may be selected from Q and N.

In the IL-2Rγc ligands of formulas (7) and (7a), X ¹¹¹ can be selected from amino acids, X ¹¹² can be selected from I and L, and X ¹¹³ can be selected from amino acids. , X ¹¹⁴ can be selected from D, E, and S, X ¹¹⁵ can be L, X ¹¹⁶ can be selected from D and E, X ¹¹⁷ can be selected from F and W, X ¹¹⁸ can be selected from F, W, and Y, X ¹¹⁹ can be selected from F, I, and L, X ¹²⁰ can be selected from amino acids and X ¹²¹ can be selected from Q and N.

The IL-2Rγc ligand can comprise an amino acid sequence selected from any one of SEQ ID NOs:255-264 and SEQ ID NOs:923-930.

The IL-2Rγc ligand can comprise an amino acid sequence selected from any one of SEQ ID NOs:255-264 and SEQ ID NOs:923-930, wherein the amino acid sequence is on the N-terminus, on the C-terminus , or with amino acids -GG- on both the N- and C-termini.

The IL-2Rγc ligand can comprise an amino acid sequence selected from any one of SEQ ID NOs:255-264 and SEQ ID NOs:923-930, each amino acid independently having the following conservation substitution: amino acids with small hydrophobic side chains, including alanine (A), glycine (G), proline (P), serine (S), or threonine (T); serine (S), threonine (T), or Amino acids with hydroxyl-containing side chains, including tyrosine (Y); amino acids with acidic side chains, including aspartic acid (D), or glutamic acid (E); histidine (H), asparagine (N), glutamine (Q), serine. (S), threonine (T), or tyrosine (Y); amino acids with basic side chains, including arginine (R), lysine (K), or histidine (H); amino acids with large hydrophobic side chains including isoleucine (I), leucine (L), methionine (M), valine (V), phenylalanine (F), tyrosine (Y), or tryptophan (W); ), histidine (H), tryptophan (W), or tyrosine (Y).

The IL-2Rγc ligands of SEQ ID NOs:255-264 and SEQ ID NOs:923-930 exhibit binding affinities (IC ₅₀ ) for the IL-2Rγc subunit of less than 100 μM.

The IL-2Rγc ligand can comprise the amino acid sequence of formula (8) (SEQ ID NO:931),
^-CX131 -X132 ^{-X133 -X134 -X135 -X136 -X137 -X138 -X139 -X140 -X141 -X142 -C-} (8)
wherein X ¹³¹ can be selected from amino acids containing large hydrophobic side chains, X ¹³² can be selected from amino acids containing large hydrophobic side chains, and X ¹³³ can be selected from amino acids containing large hydrophobic side chains. X ¹³⁴ can be selected from amino acids containing a large hydrophobic or aromatic side chain, X ¹³⁵ can be selected from amino acids containing a basic side chain as well as an acidic or polar neutral side chain X ¹³⁶ can be selected from amino acids, X ¹³⁷ can be selected from amino acids containing small hydrophobic side chains, X ¹³⁸ can be selected from acidic or polar neutral can be selected from amino acids containing side chains, X ¹³⁹ can be selected from amino acids containing large hydrophobic or aromatic side chains, and X ¹⁴⁰ can be selected from amino acids containing small hydrophobic or hydroxyl-containing side chains X ¹⁴¹ can be selected from amino acids containing large hydrophobic or aromatic side chains, and X ¹⁴² can be selected from amino acids containing large hydrophobic side chains can be done.

In the IL-2Rγc ligands of formula (8), X ¹³¹ can be selected from F, I, L, M, V, Y, and W, and X ¹³² can be F, I, L, M, V, Y, and W, X ¹³³ can be selected from F, I, M, V, Y, and W, X ¹³⁴ can be selected from F, H, I, L, M, V, Y and W, X ¹³⁵ can be selected from R, K, H, D, E, N, and Q, X ¹³⁶ can be selected from amino acids, X ¹³⁷ can be selected from A, G, P, S, and T; X ¹³⁸ can be selected from D, E, N, and Q; X ¹³⁹ can be selected from F, H, I, L, M, V, Y, and W, X ¹⁴⁰ can be selected from A, G, P, S, T, and Y, X ¹⁴¹ can be selected from F, H, I, L, M, V, Y, and W can be selected, and X ¹⁴² can be selected from F, I, L, M, V, Y, and W;

In the IL-2Rγc ligand of formula (8), X ¹³¹ can be selected from F and Y, X ¹³² can be I, X ¹³³ can be F, I, L, M, V, Y and W, X ¹³⁴ can be Y, X ¹³⁵ can be R, X ¹³⁶ can be selected from amino acids, X ¹³⁷ can be G X ¹³⁸ can be E, X ¹³⁹ can be F, X ¹⁴⁰ can be selected from S, T, and Y, X ¹⁴¹ can be Y and X ¹⁴² can be selected from F, I, L, M, V, Y, and W.

In IL-2Rγc ligands of formula (8), X ¹³¹ may be selected from F and Y.

In IL-2Rγc ligands of formula (8), X ¹³² may be selected from I, V, and L.

X ¹³² can be I in IL-2Rγc ligands of formula (8).

In IL-2Rγc ligands of formula (8), X ¹³³ may be selected from M, L, Y, and I.

X ¹³⁴ may be selected from F, H, and Y in the IL-2Rγc ligands of formula (8).

In IL-2Rγc ligands of formula (8), X ¹³⁴ can be Y.

In IL-2Rγc ligands of formula (8), X ¹³⁵ may be selected from R, K, D, and E.

In IL-2Rγc ligands of formula (8), X ¹³⁵ can be R.

In IL-2Rγc ligands of formula (8), X ¹³⁶ may be selected from amino acids.

X ¹³⁷ can be G in the IL-2Rγc ligand of formula (8).

X ¹³⁸ may be selected from D and E in IL-2Rγc ligands of formula (8).

X ¹³⁸ can be E in IL-2Rγc ligands of formula (8).

In IL-2Rγc ligands of formula (8), X ¹³⁹ may be selected from F, Y, and W.

X ¹³⁹ can be F in IL-2Rγc ligands of formula (8).

In IL-2Rγc ligands of formula (8), X ¹⁴⁰ may be selected from S and T.

In IL-2Rγc ligands of formula (8), X ¹⁴¹ may be selected from F, I, L, M, V, Y, and W.

In IL-2Rγc ligands of formula (8), X ¹⁴¹ can be Y.

In IL-2Rγc ligands of formula (8), X ¹⁴² may be selected from I, L, M, V, and Y.

In the IL-2Rγc ligands of formula (8), X ¹³¹ can be selected from F and Y, X ¹³² can be I, and X ¹³³ is selected from M, L, Y, and I. X ¹³⁴ can be Y, X ¹³⁵ can be R, X ¹³⁶ can be selected from amino acids, X ¹³⁷ can be G, X ¹³⁸ can be E, X ¹³⁹ can be F, X ¹⁴⁰ can be selected from S and T, X ¹⁴¹ can be Y, X ¹⁴² can be , F, I, L, M, V, Y, and W.

In the IL-2Rγc ligands of formula (8), X ¹³¹ can be selected from A, C, D, E, F, G, L, P, and Y, and X ¹³² can be C, I, L, N, S, and V, X ¹³³ can be selected from A, I, L, M, Q, R, and Y; X ¹³⁴ can be selected from F, H, K, L, T, and Y; X ¹³⁵ can be selected from D ^, E, G, H, I, K, L, P, Q, R, S, and Y; E, F, G, H, I, L, N, Q, R, S, and T, and X ¹³⁷ is C, D, E, G, K, N, P, Q, and T, X ¹³⁸ can be selected from D, E, F, K, P, R, and T; X ¹³⁹ can be selected from A, F, L, R, T, V, W , and Y, X ¹⁴⁰ can be selected from D, E, G, L, N, T, W, and Y, and X ¹⁴¹ can be selected from A, C, F, G, I , M, and Y, and X ¹⁴² may be selected from C, E, I, L, M, V, and Y.

In the IL-2Rγc ligands of formula (8), X ¹³¹ can be selected from F and Y, X ¹³² can be selected from I, L and V, X ¹³³ can be I, M, R, and Y, X ¹³⁴ can be selected from F, H, and Y, X ¹³⁵ can be selected from D, E, K, and R, and X ¹³⁶ can be selected from , E, F, G, H, I, L, N, Q, R, S, and T, X ¹³⁷ can be G, and X ¹³⁸ can be selected from D and E X ¹³⁹ can be selected from F, W, and Y; X ¹⁴⁰ can be selected from S and T; X ¹⁴¹ can be selected from F, I, L, M, and Y and X ¹⁴² can be selected from I, L, M, V, and Y.

In the IL-2Rγc ligand of formula (8), X ¹³¹ can be F, X ¹³² can be I, X ¹³³ can be selected from I, M, R, and Y; ¹³⁴ can be Y, X ¹³⁵ can be selected from D, E, K, and R, X ¹³⁶ can be E, F, G, H, I, L, N, Q, R, S, and T, X ¹³⁷ can be G, X ¹³⁸ can be E, X ¹³⁹ can be F, X ¹⁴⁰ can be selected from S and T X ¹⁴¹ can be Y and X ¹⁴² can be selected from I, L, M, V, and Y.

In IL-2Rγc ligands of formula (8), X ¹³¹ can be F, X ¹³² can be I, X ¹³⁴ can be Y and X ¹²⁵ can be R , X ¹³⁷ can be G, X ¹³⁸ can be E, X ¹³⁹ can be F, and X ¹⁴¹ can be Y.

The IL-2Rγc ligand can comprise an amino acid sequence selected from any one of SEQ ID NOs:265-267 and SEQ ID NOs:932-940.

The IL-2Rγc ligand can comprise an amino acid sequence selected from any one of SEQ ID NOs:265-267 and SEQ ID NOs:931-940, wherein the amino acid sequence is on the N-terminus, on the C-terminus , or with amino acids -GG- on both the N- and C-termini.

The IL-2Rγc ligand can comprise an amino acid sequence selected from any one of SEQ ID NOs:265-267 and SEQ ID NOs:931-940, each amino acid independently having the following conservation substitution: amino acids with small hydrophobic side chains, including alanine (A), glycine (G), proline (P), serine (S), or threonine (T); serine (S), threonine (T), or Amino acids with hydroxyl-containing side chains, including tyrosine (Y); amino acids with acidic side chains, including aspartic acid (D), or glutamic acid (E); histidine (H), asparagine (N), glutamine (Q), serine. (S), threonine (T), or tyrosine (Y); amino acids with basic side chains, including arginine (R), lysine (K), or histidine (H); amino acids with large hydrophobic side chains including isoleucine (I), leucine (L), methionine (M), valine (V), phenylalanine (F), tyrosine (Y), or tryptophan (W); ), histidine (H), tryptophan (W), or tyrosine (Y).

The IL-2Rγc ligands of SEQ ID NOs:265-267 and SEQ ID NOs:931-940 exhibit binding affinities (IC ₅₀ ) for the IL-2Rγc subunit of less than 100 μM.

The IL-2Rγc ligand can comprise the amino acid sequence of formula (9) (SEQ ID NO:941) or the amino acid sequence of formula (9a) (SEQ ID NO:942),
^{-X155 -X156 -X157 -X158 -X159} - (9)
^{-X151 -X152 -X153 -X154 -CX155} -X156 ^{-X157 -X158 -X159 -CX160 -X161 -X162 -X163 -} (9a)
wherein X ¹⁵¹ can be selected from amino acids containing small hydrophobic or hydroxyl-containing side chains, X ¹⁵² can be selected from amino acids containing large hydrophobic side chains, and X ¹⁵³ is Can be selected from amino acids containing acidic or polar neutral side chains, X ¹⁵⁴ can be selected from amino acids containing basic side chains, and X ¹⁵⁵ can be selected from amino acids containing large hydrophobic side chains. X ¹⁵⁶ can be selected from amino acids containing small hydrophobic or hydroxyl-containing side chains, X ¹⁵⁷ can be selected from amino acids containing small hydrophobic side chains, and X ¹⁵⁸ can be selected from , can be selected from amino acids containing small hydrophobic or hydroxyl-containing side chains, X ¹⁵⁹ can be selected from amino acids containing small hydrophobic or hydroxyl-containing side chains, and X ¹⁶⁰ can be selected from amino acids containing small can be selected from amino acids containing hydrophobic or hydroxyl-containing side chains, X ¹⁶¹ can be selected from amino acids and X ¹⁶² can be selected from amino acids containing large hydrophobic or basic side chains and X ¹⁶³ can be selected from amino acids containing large hydrophobic side chains.

In the IL-2Rγc ligands of Formula (9) and Formula (9a), X ¹⁵¹ can be selected from A, G, P, S, and T, and X ¹⁵² can be F, I, L, M, V , Y, and W; X ¹⁵³ may be selected from D, E, N, and Q; X ¹⁵⁴ may be selected from H ^, K, and R; can be selected from F, I, L, M, V, Y, and W, X ¹⁵⁶ can be selected from A, G, P, S, and T, and X ¹⁵⁷ can be selected from A, G, P, S, and T, X ¹⁵⁸ can be selected from A, G, P, S, and T, X ¹⁵⁹ can be selected from A, G, P, S, and T X ¹⁶⁰ can be selected from A, G, P, S, and T; X ¹⁶¹ can be selected from amino acids; X ¹⁶² can be selected from F, I, L, M, V, Y, W, K, and H, and X ¹⁶³ can be selected from F, I, L, M, V, Y, and W.

In the IL-2Rγc ligands of Formula (9) and Formula (9a), X ¹⁵¹ can be selected from K, M, N, and K, and X ¹⁵² can be selected from M, L, and Y. X ¹⁵³ can be selected from N, Y, and L, X ¹⁵⁴ can be K, and X ¹⁵⁵ can be selected from A, W, R, Y, and N , X ¹⁵⁶ can be selected from T, N, and S, X ¹⁵⁷ can be selected from P and A, and X ¹⁵⁸ can be selected from S, R, F, and L. , X ¹⁵⁹ can be selected from Q, S, E, and T, X ¹⁶⁰ can be selected from S, Q, and A, and X ¹⁶¹ can be selected from V, S, G, L, and N, X ¹⁶² can be selected from I, K, R, and V, and X ¹⁶³ can be selected from F and L.

In IL-2Rγc ligands of formula (9) and formula (9a), X ¹⁵¹ may be selected from S and T.

In IL-2Rγc ligands of formula (9) and formula (9a), X ¹⁵² may be selected from L and M.

X ¹⁵² can be L in IL-2Rγc ligands of formula (9) and formula (9a).

X ¹⁵³ can be N in IL-2Rγc ligands of formula (9) and formula (9a).

X ¹⁵⁴ can be K in IL-2Rγc ligands of formula (9) and formula (9a).

In IL-2Rγc ligands of formula (9) and formula (9a), X ¹⁵⁵ may be selected from W and Y.

In IL-2Rγc ligands of formula (9) and formula (9a), X ¹⁵⁶ may be selected from S and T.

X ¹⁵⁶ can be S in IL-2Rγc ligands of formula (9) and formula (9a).

X ¹⁵⁷ can be P in IL-2Rγc ligands of formula (9) and formula (9a).

X ¹⁵⁸ can be S in IL-2Rγc ligands of formula (9) and formula (9a).

In IL-2Rγc ligands of formula (9) and formula (9a), X ¹⁵⁹ may be selected from S and T.

X ¹⁵⁹ can be S in IL-2Rγc ligands of formula (9) and formula (9a).

X ¹⁶⁰ can be S in IL-2Rγc ligands of formula (9) and formula (9a).

In IL-2Rγc ligands of formula (9) and formula (9a), X ¹⁶¹ may be selected from amino acids.

In IL-2Rγc ligands of formula (9) and formula (9a), X ¹⁶² may be selected from I, V, R, and K.

In IL-2Rγc ligands of formula (9) and formula (9a), X ¹⁶² may be selected from I and V.

In IL-2Rγc ligands of formula (9) and formula (9a), X ¹⁶² may be selected from R and K.

In IL-2Rγc ligands of formula (9) and formula (9a), X ¹⁶³ may be selected from F and L.

X ¹⁶³ can be L in IL-2Rγc ligands of formula (9) and formula (9a).

In the IL-2Rγc ligands of formula (9) and formula (9a), X ¹⁵¹ can be selected from S and T, X ¹⁵² can be L and X ¹⁵³ can be N X ¹⁵⁴ can be K, X ¹⁵⁵ can be selected from W and Y, X ¹⁵⁶ can be S, X ¹⁵⁷ can be P, X ¹⁵⁸ can be S, X ¹⁵⁹ can be S, X ¹⁶⁰ can be S, T, X ¹⁶¹ can be selected from amino acids, X ¹⁶² can be can be I and X ¹⁶³ can be F;

In the IL-2Rγc ligands of formula (9) and formula (9a), X ¹⁵² can be L, X ¹⁵³ can be N, X ¹⁵⁴ can be K, X ¹⁵⁶ can be S X ¹⁵⁷ can be P, X ¹⁵⁸ can be S, X ¹⁵⁹ can be S, X ¹⁶⁰ can be S, X ¹⁶² can be I and X ¹⁶³ can be F.

IL-2Rγc ligands comprise an amino acid sequence selected from any one of SEQ ID NOs:943-948.

The IL-2Rγc ligand can comprise an amino acid sequence selected from any one of SEQ ID NOs:941-948, wherein the amino acid sequence is on the N-terminus, the C-terminus, or both the N-terminus and the C-terminus. It may end with the above amino acids -GG-.

The IL-2Rγc ligand may comprise an amino acid sequence selected from any one of SEQ ID NOs:941-948, each amino acid independently having the following conservative substitutions: alanine (A), glycine (G), amino acids with small hydrophobic side chains including proline (P), serine (S), or threonine (T); hydroxyl-containing sides including serine (S), threonine (T), or tyrosine (Y) amino acids with acidic side chains, including aspartic acid (D), or glutamic acid (E); histidine (H), asparagine (N), glutamine (Q), serine (S), threonine (T), or amino acids with polar neutral side chains, including tyrosine (Y); amino acids with basic side chains, including arginine (R), lysine (K), or histidine (H); isoleucine (I), leucine (L) , methionine (M), valine (V), phenylalanine (F), tyrosine (Y), or tryptophan (W); and phenylalanine (F), histidine (H), tryptophan ( W), or amino acids with aromatic side chains, including tyrosine (Y).

The IL-2Rγc ligands of SEQ ID NOs:941-948 exhibit binding affinities (IC ₅₀ ) for the IL-2Rγc subunit of less than 100 μM.

The IL-2Rγc ligand can comprise the amino acid sequence of formula (12) (SEQ ID NO:949),
-X ¹⁷¹ -X ¹⁷² -X ¹⁷³ -X ¹⁷⁴ -X ¹⁷⁵ -CX ^{176 -X} 177 ^-X 178 -X ¹⁷⁹ -X ¹⁸⁰ -X ¹⁸¹ -X ¹⁸² -X ¹⁸³ -CX ¹⁸⁴ -X ¹⁸⁵ - X ¹⁸⁶ -X ¹⁸⁷ -X ¹⁸⁸ - (12)
wherein X ¹⁷¹ can be selected from amino acids containing basic side chains, X ¹⁷² can be selected from amino acids containing hydroxyl-containing side chains, and X ¹⁷³ can be selected from acidic side chains or large hydrophobic Can be selected from amino acids containing a side chain, X ¹⁷⁴ can be selected from amino acids containing a large hydrophobic side chain, and X ¹⁷⁵ can be selected from amino acids containing an acidic side chain or a large hydrophobic side chain. X ¹⁷⁶ can be selected from amino acids containing acidic or polar/neutral side chains, X ¹⁷⁷ can be selected from amino acids containing acidic side chains, and X ¹⁷⁸ can be selected from large can be selected from amino acids containing hydrophobic or aromatic side chains, X ¹⁷⁹ can be selected from amino acids containing acidic or polar/neutral side chains, and X ¹⁸⁰ is G can be, X ¹⁸¹ can be V, X ¹⁸² can be E, X ¹⁸³ can be L, X ¹⁸⁴ can be W, X ¹⁸⁵ can be a large hydrophobic can be selected from amino acids containing side chains, X ¹⁸⁶ can be E, X ¹⁸⁷ can be selected from amino acids, and X ¹⁸⁸ can be selected from amino acids containing acidic side chains .

In the IL-2Rγc ligands of formula (12), X ¹⁷¹ can be selected from H, K and R, X ¹⁷² can be selected from S, T and Y, X ¹⁷³ can be selected from D , E, F, I, L, M, V, W, and Y; X ¹⁷⁴ may be selected from F, I, L, M, V, W, and Y; ¹⁷⁵ can be selected from D, E, F, I, L, M, V, W, and Y; X ¹⁷⁶ can be selected from D, E, H, N, Q, S, T, and Y; and X ¹⁷⁷ can be selected from D and E, X ¹⁷⁸ can be selected from F, H, I, L, M, V, W, and Y, and X ¹⁷⁹ can be selected from D, E, H, N, Q, S, T, and Y, X ¹⁸⁰ can be G, X ¹⁸¹ can be V, X ¹⁸² can be E , X ¹⁸³ can be L, X ¹⁸⁴ can be W, and X ¹⁸⁵ can be selected from F, I, L, M, V, W, and Y X ¹⁸⁶ can be E, X ¹⁸⁷ can be selected from amino acids and X ¹⁸⁸ can be selected from D and E.

In the IL-2Rγc ligands of formula (12), X ¹⁷¹ can be selected from D, E, G, H, K, M, N, P, Q, R, S, and T, and X ¹⁷² is A, D, E, G, I, K, L, P, Q, R, S, T, V, W, and Y, and X ¹⁷³ is A, D, E, F, G , I, Q, S, T, V, W, and Y, and X ¹⁷⁴ can be selected from A, I, E, I, L, M, N, Q, R, S, T, and V X ¹⁷⁵ can be selected from A, E, I, L, M, N, Q, R, S, T, and V; X ¹⁷⁶ can be selected from D, E, H, L, Q, R, and V can be selected, X ¹⁷⁷ can be selected from D, E, N, T, and V, and X ¹⁷⁸ can be selected from F, S, W, and Y X ¹⁷⁹ can be selected from A, D, E, G, H, K, N, Q, R, and Y; X ¹⁸⁰ can be selected from G and R; X ¹⁸¹ can be V, X ¹⁸² can be selected from D, E, and Y, X ¹⁸³ can be selected from F, I, and L, and X ¹⁸⁴ is W X ¹⁸⁵ can be selected from C, H, I, L, P, Q, T, V, and Y; X ¹⁸⁶ can be selected from A, D, E, G, M, R, S , T, and V, and X ¹⁸⁷ is selected from A, D, E, F, G, I, M, N, P, Q, R, S, T, V, W, and Y and X ¹⁸⁸ can be selected from A, C, D, E, F, G, I, K, L, N, P, Q, R, S, and V.

In the IL-2Rγc ligands of formula (12), X ¹⁷¹ can be selected from H, K and R, X ¹⁷² can be selected from S, T and Y, X ¹⁷³ can be selected from D , E, F, I, and V, X ¹⁷⁴ can be selected from I and V, and X ¹⁷⁵ can be selected from E, I, L, M, and V. , X ¹⁷⁶ can be selected from D, E, and Q, X ¹⁷⁷ can be selected from D and E, X ¹⁷⁸ can be selected from F and W, and X ¹⁷⁹ can be selected from D, E, N, and Q, X ¹⁸⁰ can be G, X ¹⁸¹ can be V, and X ¹⁸² can be selected from D and E , X ¹⁸³ can be L, X ¹⁸⁴ can be W, X ¹⁸⁵ can be selected from I, L, Q, and V, and X ¹⁸⁶ can be from D and E X ¹⁸⁷ can be selected from A, D, E, F, G, I, M, N, P, Q, R, S, T, V, and W, and Y; X ¹⁸⁸ can be selected from D, E, N, and Q.

In the IL-2Rγc ligands of formula (12), X ¹⁷¹ can be selected from K and R, X ¹⁷² can be selected from S, T and Y, X ¹⁷³ can be D, E, F, I, and V can be selected, X ¹⁷⁴ can be V, X ¹⁷⁵ can be selected from E, L, M, and V, and X ¹⁷⁶ can be Q X ¹⁷⁷ can be selected from D and E; X ¹⁷⁸ can be W; can be selected ^from X ¹⁷⁹ , D, E, N, and Q; G, X ¹⁸¹ can be V, X ¹⁸² can be E, X ¹⁸³ can be L, X ¹⁸⁴ can be W , X ¹⁸⁵ can be selected from I, L, Q, and V, X ¹⁸⁶ can be selected from D and E, and X ¹⁸⁷ can be selected from A, D, E, F, G, I, M, N, P, Q, R, S, T, V, W, and Y can be selected, and X ¹⁸⁸ can be selected from D, E, N, and Q.

In IL-2Rγc ligands of formula (12), X ¹⁷¹ may be selected from H, K, and R.

In IL-2Rγc ligands of formula (12), X ¹⁷² may be selected from S, T, and Y.

In IL-2Rγc ligands of formula (12), X ¹⁷³ may be selected from D, E, F, I, L, M, V, W, and Y.

X ¹⁷³ may be selected from D and E in IL-2Rγc ligands of formula (12).

In IL-2Rγc ligands of formula (12), X ¹⁷³ may be selected from F, I, L, M, V, W, and Y.

In IL-2Rγc ligands of formula (12), X ¹⁷⁴ may be selected from F, I, L, M, V, W, and Y.

X ¹⁷⁴ can be V in the IL-2Rγc ligand of formula (12).

In IL-2Rγc ligands of formula (12), X ¹⁷⁵ may be selected from D, E, F, I, L, M, V, W, and Y.

X ¹⁷⁵ may be selected from D and E in IL-2Rγc ligands of formula (12).

In IL-2Rγc ligands of formula (12), X ¹⁷⁵ may be selected from F, I, L, M, V, W, and Y.

In IL-2Rγc ligands of formula (12), X ¹⁷⁶ may be selected from D, E, H, N, Q, S, T, and Y.

In IL-2Rγc ligands of formula (12), X ¹⁷⁶ may be selected from E and Q.

X ¹⁷⁷ may be selected from D and E in IL-2Rγc ligands of formula (12).

In IL-2Rγc ligands of formula (12), X ¹⁷⁸ can be selected from F, H, I, L, M, V, W, and Y.

In IL-2Rγc ligands of formula (12), X ¹⁷⁸ may be selected from F, H, W, and Y.

X ¹⁷⁸ can be W in IL-2Rγc ligands of formula (12).

In IL-2Rγc ligands of formula (12), X ¹⁷⁹ may be selected from D, E, H, N, Q, S, T, and Y.

X ¹⁷⁹ may be selected from D, E, and Q in IL-2Rγc ligands of formula (12).

X ¹⁸⁰ can be G in the IL-2Rγc ligand of formula (12).

X ¹⁸¹ can be V in the IL-2Rγc ligand of formula (12).

X ¹⁸² can be E in IL-2Rγc ligands of formula (12).

X ¹⁸³ can be L in IL-2Rγc ligands of formula (12).

In IL-2Rγc ligands of formula (12), X ¹⁸⁴ can be W.

In IL-2Rγc ligands of formula (12), X ¹⁸⁵ may be selected from F, I, L, M, V, W, and Y.

X ¹⁸⁵ can be L in IL-2Rγc ligands of formula (12).

X ¹⁸⁶ can be E in IL-2Rγc ligands of formula (12).

In IL-2Rγc ligands of formula (12), X ¹⁸⁷ may be selected from amino acids.

X ¹⁸⁸ may be selected from D and E in IL-2Rγc ligands of formula (12).

In the IL-2Rγc ligands of formula (12), X ¹⁷¹ can be selected from H, K and R, X ¹⁷² can be selected from S, T and Y, X ¹⁷³ can be selected from D , E, F, I, L, M, V, W, and Y; X ¹⁷⁴ may be selected from F, I, L, M, V, W, and Y; ¹⁷⁵ can be selected from D, E, F, I, L, M, V, W, and Y; X ¹⁷⁶ can be selected from D, E, H, N, Q, S, T, and Y; and X ¹⁷⁷ can be selected from D and E, X ¹⁷⁸ can be selected from F, H, I, L, M, V, W, and Y, and X ¹⁷⁹ can be selected from D, E, H, N, Q, S, T, and Y, X ¹⁸⁰ can be G, X ¹⁸¹ can be V, X ¹⁸² can be E , X ¹⁸³ can be L, X ¹⁸⁴ can be selected from W, and X ¹⁸⁵ can be selected from F, I, L, M, V, W, and Y X ¹⁸⁶ can be E, X ¹⁸⁷ can be selected from amino acids and X ¹⁸⁸ can be selected from D and E.

In the IL-2Rγc ligands of formula (12), X ¹⁷¹ can be selected from H, K and R, X ¹⁷² can be selected from S, T and Y, X ¹⁷³ can be selected from D and E, X ¹⁷⁴ can be V, X ¹⁷⁵ can be selected from D and E, X ¹⁷⁶ can be selected from E and Q, X ¹⁷⁷ can be selected from D ^and E; X ¹⁷⁸ can be selected from F, H, W, and Y; X ¹⁷⁹ can be selected from D, E, and Q; can be G, X ¹⁸¹ can be V, X ¹⁸² can be E, X ¹⁸³ can be L, X ¹⁸⁴ can be W X ¹⁸⁵ can be selected from F, I, L, M, V, W, and Y, X ¹⁸⁶ can be E, and X ¹⁸⁷ can be selected from amino acids , X ¹⁸⁸ can be selected from D and E.

In the IL-2Rγc ligands of formula (12), X ¹⁷¹ can be selected from H, K and R, X ¹⁷² can be selected from S, T and Y, X ¹⁷³ can be selected from F , I, L, M, V, W, and Y, X ¹⁷⁴ can be V, and X ¹⁷⁵ can be selected from F, I, L, M, V, W, and Y X ¹⁷⁶ can be selected from E and Q, X ¹⁷⁷ can be selected from D and E, X ¹⁷⁸ can be selected from F, H, W, and Y and X ¹⁷⁹ can be selected from D, E, and Q, X ¹⁸⁰ can be G, X ¹⁸¹ can be V, and X ¹⁸² can be E can be, X ¹⁸³ can be L, X ¹⁸⁴ can be W, X ¹⁸⁵ can be selected from F, I, L, M, V, W, and Y; X ¹⁸⁶ can be E, X ¹⁸⁷ can be selected from amino acids and X ¹⁸⁸ can be selected from D and E.

In IL-2Rγc ligands of formula (12), X ¹⁷¹ can be selected from H, K, and R, X ¹⁷² can be selected from S, T, and Y, and X ¹⁷³ can be selected from D , E, F, I, L, M, V, W, and Y, X ¹⁷⁴ can be V, X ¹⁷⁵ can be D, E, F, I, L, M , V, W, and Y, X ¹⁷⁶ can be selected from D, E, H, N, Q, S, T, and Y, and X ¹⁷⁶ can be selected from E and Q , X ¹⁷⁷ can be selected from D and E, X ¹⁷⁸ can be W, X ¹⁷⁹ can be selected from D, E, and Q, and X ¹⁸⁰ can be , G, X ¹⁸¹ can be V, X ¹⁸² can be E, X ¹⁸³ can be L, X ¹⁸⁴ can be W X ¹⁸⁵ can be selected from F, I, L, M, V, W, and Y; X ¹⁸⁶ can be E; X ¹⁸⁷ can be selected from amino acids; X ¹⁸⁸ can be selected from D and E;

IL-2Rγc ligands comprise an amino acid sequence selected from any one of SEQ ID NO:950-SEQ ID NO:1027.

The IL-2Rγc ligand can comprise an amino acid sequence selected from any one of SEQ ID NO:949 to SEQ ID NO:1027, wherein the amino acid sequence is on the N-terminus, the C-terminus, or both the N-terminus and the C-terminus. It may end with the above amino acids -GG-.

The IL-2Rγc ligand may comprise an amino acid sequence selected from any one of SEQ ID NOs:949-1027, each amino acid independently having the following conservative substitutions: alanine (A), glycine (G), amino acids with small hydrophobic side chains including proline (P), serine (S), or threonine (T); hydroxyl-containing sides including serine (S), threonine (T), or tyrosine (Y) amino acids with acidic side chains, including aspartic acid (D), or glutamic acid (E); histidine (H), asparagine (N), glutamine (Q), serine (S), threonine (T), or amino acids with polar neutral side chains, including tyrosine (Y); amino acids with basic side chains, including arginine (R), lysine (K), or histidine (H); isoleucine (I), leucine (L) , methionine (M), valine (V), phenylalanine (F), tyrosine (Y), or tryptophan (W); and phenylalanine (F), histidine (H), tryptophan ( W), or amino acids with aromatic side chains, including tyrosine (Y).

The IL-2Rγc ligands of SEQ ID NOs:949-1027 exhibit binding affinities (IC ₅₀ ) for the IL-2Rγc subunit of less than 100 μM.

IL-2Rγc ligands can include, for example, 5-50 amino acids, 5-40 amino acids, 5-30 amino acids, 5-30 amino acids, 6-25 amino acids, or 7-20 amino acids.

IL-2Rγc ligand is 1 pM to 100 μM, 10 pM to 10 μM, to human IL-2Rγc subunit, to mammalian IL-2Rγc subunit, or to both human IL-2Rγc subunit and mammalian IL-2Rγc subunit; Binding affinities (IC ₅₀ ) of 100 pM to 1 μM, 0.001 μM to 1 μM, or 0.01 μM to 1 μM can be exhibited.

IL-2Rγc ligands provided by the present disclosure can exhibit binding affinities (IC ₅₀ ) for human IL-2Rγc subunits of, for example, 0.1 μM to 50 μM.

IL-2Rγc ligand is less than 100 μM, less than 10 μM, less than 1 μM to human IL-2Rγc subunit, to mammalian IL-2Rγc subunit, or to both human IL-2Rγc subunit and mammalian IL-2Rγc subunit , less than 0.1 μM, or less than 0.01 μM binding affinities (IC ₅₀ ).

The IL-2Rγc ligand has a binding affinity (IC ₅₀ ) of less than 100 μM, less than 10 μM, less than 1 μM, less than 0.1 μM, or less than 0.01 μM for human IL-2Rγc subunit and human IL-2Rγc subunit, respectively can be shown.

The IL-2Rγc ligand has a binding affinity of 1 pM to 100 μM, 10 pM to 10 μM, 100 pM to 1 μM, 0.001 μM to 1 μM, or 0.01 μM to 1 μM for human IL-2Rγc subunit and human IL-2Rγc subunit, respectively. properties ( _IC50 ).

IL-2Rγc ligands provided by the present disclosure can exhibit a binding affinity (IC ₅₀ ) for the human IL-2Rα (CD25) subunit of greater than 100 μM, greater than 1 mM, greater than 10 mM, or greater than 100 mM.

The IL-2Rγc ligand is at least 10-fold greater, at least 50-fold greater, at least 100-fold greater, at least 500-fold greater, or at least 1-fold greater than the binding affinity (IC ₅₀ ) of the IL-2Rγc ligand for the human IL-2Rα subunit. ,000-fold greater binding affinity (IC ₅₀ ) for the human IL-2Rγc subunit.

Amino acid sequences having SEQ ID NO:268 to SEQ ID NO:376 are excluded from the scope of amino acid sequences according to the present invention. In other respects, in the genus or subgenus of amino acid sequences encompassing any one of SEQ ID NO:268-SEQ ID NO:376, the amino acid sequence of SEQ ID NO:268-SEQ ID NO:376 covers the genus or subgenus not included in

Solid tumors exhibit metabolic differences from normal tissue. The greater reliance of solid tumors on glycolytic metabolism produces a more acidic tumor microenvironment. For example, a solid tumor microenvironment can have a pH that is 1-2 pH lower than that of most normal tissues. This pH difference can be used to enhance the activity of therapeutic agents in solid tumors relative to their activity in normal peripheral tissues.

Suitable pH-selective screening methods can be used to identify peptides with greater binding affinity for IL-2 at lower pH and weaker binding affinity at neutral pH. IL-2 agonists and antagonists can be constructed based on the identified pH-selective peptides. These pH-selective agonists and antagonists can exhibit enhanced therapeutic indices reflecting increased cytotoxicity targeting solid tumors and reduced toxicity to normal tissues.

Acid-biased affinity selection showed receptor ligands with increased affinity at pH<6.5, comparable to solid tumor microenvironment, and decreased affinity at neutral pH>7.0, comparable to normal tissue affinity. is used to identify These peptide ligands can serve as building blocks for constructing pH-targeted and pH-selective antagonists and agonists.

IL-2Rβ ligands provided by this disclosure can include pH-selective IL-2Rβ ligands.

pH-selective IL-2Rβ ligands comprise the amino acid sequence of formula (1) (SEQ ID NO:1), the amino acid sequence of formula (1a) (SEQ ID NO:2), or the amino acid sequence of formula (1b) (SEQ ID NO:3). get,
^{-X3 -X4 -X5 -X6 -X7 -X8 -X9 -X10} - (1)
^-X2 -CX3 ^{-X4 -X5 -X6 -X7 -X8 -X9 -X10 -CX11} - ( ^1a )
^{-X1 -X2} -CX3 ^{-X4 -X5 -X6 -X7 -X8 -X9 -X10 -CX11 -X12 -} (1b)
wherein X ¹ can be selected from amino acids containing large hydrophobic side chains, X ² can be selected from amino acids, X ³ can be selected from amino acids, and X ⁴ can be selected from can be selected from amino acids, X ⁵ can be selected from amino acids, X ⁶ can be selected from amino acids, X ⁷ can be selected from amino acids, X ⁸ can be selected from can be selected from amino acids comprising a polar or acidic side chain, X ⁹ can be selected from amino acids comprising a polar neutral or acidic side chain, X ¹⁰ can be selected from amino acids, X ¹¹ can be selected from amino acids and X ¹² can be selected from amino acids.

In the pH-selective IL-2Rβ ligands of formulas (1)-(1b), X ¹ can be selected from amino acids containing large hydrophobic side chains and X ² is selected from amino acids containing acidic side chains. X ³ can be selected from amino acids, X ⁴ can be selected from amino acids, X ⁵ can be selected from amino acids containing small hydrophobic side chains, X ⁶ can be selected from , can be selected from amino acids containing polar neutral or acidic side chains, X ⁷ can be selected from amino acids containing polar neutral or large hydrophobic side chains, and X ⁸ can be selected from amino acids containing small hydrophobic side chains X ⁹ can be selected from amino acids containing polar neutral or acidic side chains, X ¹⁰ can be selected from amino acids containing large hydrophobic side chains, X ¹¹ can be selected from amino acids containing acidic side chains and X ¹² can be selected from amino acids containing large hydrophobic side chains.

In the pH-selective IL-2Rβ ligands of formulas (1)-(1b), X ¹ can be selected from I, L, M, ^V , F, W, and Y; X ³ can be selected from amino acids, X ⁴ can be selected from amino acids, X ⁵ can be selected from A, G, P, S, and T X ⁶ can be selected from H, N, Q, S, T, Y, D, and E; X ⁷ can be selected from H, N, Q, S, T, Y, I, L, M , V, F, W, and Y, X ⁸ can be selected from A, G, P, S, and T, X ⁹ can be selected from H, N, Q, S, T , Y, D, and E; X ¹⁰ may be selected from I, L, M, V, F, W, and Y; X ¹¹ may be selected from D and E; and X ¹² can be selected from I, L, M, V, F, W, and Y.

In the pH-selective IL-2Rβ ligands of formulas (1)-(1b), X ¹ can be selected from I, L, M, V ^, F, W, and Y; X ³ can be selected from amino acids, X ⁴ can be selected from amino acids, X ⁵ can be A, X ⁶ can be H, N, Q, S, T, Y, D, and E, and X ⁷ from H, N, Q, S, T, Y, I, L, M, V, F, W, and Y X ⁸ can be G; X ⁹ can be selected from H, N, Q, S, T, Y, D, and E; X ¹⁰ can be selected from I, L, M, V, F, W, and Y, X ¹¹ can be selected from D and E, and X ¹² can be selected from I, L, M, V, F, W, and Y.

In the pH-selective IL-2Rβ ligands of formulas (1)-(1b), ^X1 may be selected from L, I, F and V.

In the pH-selective IL-2Rβ ligands of formulas (1)-(1b), ^X2 may be selected from D and E.

In the pH-selective IL-2Rβ ligands of formulas (1)-(1b), ^X6 may be selected from Q, E, and D.

^X7 may be selected from V, L, and I in the pH-selective IL-2Rβ ligands of formulas (1)-(1b).

In the pH-selective IL-2Rβ ligands of formulas (1)-(1b), ^X9 may be selected from E, D, and Q.

X ¹⁰ may be selected from L, V, I, and Y in the pH-selective IL-2Rβ ligands of formulas (1)-(1b).

In the pH-selective IL-2Rβ ligands of formulas (1)-(1b), X ¹¹ may be selected from D and E.

In pH-selective IL-2Rβ ligands of formulas (1)-(1b), X ¹² may be selected from L, I, and F.

In the pH-selective IL-2Rβ ligands of formulas (1)-(1b), X ¹ can be selected from L, I, F, and V, and X ² can be selected from D and E. , X ⁶ can be selected from Q, E, and D, X ⁷ can be selected from V, L, and I, and X ⁹ can be selected from E, D, and Q X ¹⁰ can be selected from L, V, I, and Y, X ¹¹ can be selected from D and E, and X ¹² can be selected from L, I, and F can.

In the pH-selective IL-2Rβ ligands of formulas (1)-(1b), X ¹ can be selected from F, I, M, and Y and X ² is selected from E, D, and R X ³ can be selected from and amino acids, X ⁴ can be selected from amino acids, X ⁵ can be A, X ⁶ can be A, P, and Q X ⁷ can be selected from I and V, X ⁸ can be G, X ⁹ can be selected from E and Q, X ¹⁰ can be selected from I, L, and V can be selected, X ¹¹ can be selected from E, D, and Q, and X ¹² can be selected from I and L.

A pH-selective IL-2Rβ ligand comprises an amino acid sequence selected from any one of SEQ ID NOs:400-577.

pH-selective IL-2Rβ ligands that show a greater than 15% reduction in binding to the IL-2Rβ receptor at pH 7.5 compared to pH 6.0 include SEQ ID NOS: 400, 402-405, 407, 409, 410, 411, 413, 415, 416, 418, 419, 420, 421, 423, 425-432, 436, 438-440, 442-446, 448, 450, 452, 453-456, 459-461, 463-468, 470, 471, 473-477, 479, 481-486, 489, 491, 493-496, 498-507, 510-519, 521-524, 526-531, 534-537, 543, 545-548, 550, 551, 558-564, 566, 568-573, and 575, and pH-selective binding determined as described in Examples 9-12.

pH-selective IL-2Rβ ligands that show a greater than 50% reduction in binding to the IL-2Rβ receptor at pH 7.5 compared to pH 6.0 include SEQ ID NOS: 400, 404-405, 407, 409, 410, 413, 415, 420, 426, 431, 432, 438-440, 442, 444-446, 450, 452, 453, 455, 459, 464-467, 473-475, 479, 480, 482-484, 486, 489, 493, 496, 498, 502, 504, 510-514, 516-519, 521-523, 527-530, 537, 543, 545-547, 550, 558-564, 571-573, and 575 and pH-selective binding is determined as described in Examples 9-12.

pH-selective pH-selective IL-2Rβ ligands showed greater than 99% reduction in binding to IL-2Rβ receptors at pH 7.5 compared to pH 6.0; can exhibit a greater than 90%, greater than 80%, greater than 70%, greater than 60%, greater than 50%, greater than 40%, greater than 30%, greater than 20%, or greater than 10% reduction in binding to the IL-2Rβ receptor; pH-selective binding is determined as described in Examples 9-12.

A pH-selective pH-selective IL-2Rβ ligand can comprise 5-30 amino acids.

pH-selective pH-selective IL-2Rβ ligands can exhibit binding affinities from 1 pM to 100 μM for human IL-2Rβ subunits at pH 6.0.

pH-selective pH-selective IL-2Rβ ligands can exhibit binding affinities of 0.1 μM to 50 μM for human IL-2Rβ subunits at pH 6.0.

A pH-selective pH-selective IL-2Rβ ligand can exhibit a binding affinity for human IL-2Rβ subunits of less than 100 μM at pH 6.0.

A pH-selective pH-selective IL-2Rβ ligand can exhibit a binding affinity for mammalian IL-2Rβ subunits of less than 100 μM at pH 6.0.

A pH-selective pH-selective IL-2Rβ ligand can exhibit a binding affinity of less than 100 μM for each of the human IL-2Rβ subunit and the human IL-2Rγc subunit.

pH-selective pH-selective IL-2Rβ ligands can exhibit binding affinities greater than 100 μM for human IL-2Rα (CD25) subunits.

A pH-selective pH-selective IL-2Rβ ligand can exhibit a binding affinity for a human IL-2Rβ subunit that is at least 10-fold higher than the binding affinity of the IL-2Rβ ligand for a human IL-2Rα subunit.

Suitable pH-selective screening methods can be used to identify peptides with greater binding affinity for IL-2 at lower pH and weaker binding affinity at neutral pH. IL-2 agonists and antagonists can be constructed based on the identified pH-selective peptides. These pH-selective agonists and antagonists can exhibit enhanced therapeutic indices reflecting increased cytotoxicity targeting solid tumors and reduced toxicity to normal tissues.

Acid-biased affinity selection showed receptor ligands with increased affinity at pH<6.5, comparable to solid tumor microenvironment, and decreased affinity at neutral pH>7.0, comparable to normal tissue affinity. is used to identify These peptide ligands can serve as building blocks for constructing pH-targeted and pH-selective antagonists and agonists.

Certain IL-2Rβ ligands provided by this disclosure exhibit binding affinities for IL-2Rβ subunits of less than 10 μM and binding affinities (IC ₅₀ ) for IL-2Rγc subunits of greater than 100 μM.

Certain families of IL-2Rβ ligands bind IL-2Rβ subunits competitively with certain other families of IL-2Rβ ligands, bind non-competitively with other families of IL-2Rβ ligands, and It binds IL-2 non-competitively and has no detectable binding to the IL-2Rγc subunit.

The specific binding site of the IL-2Rβ subunit is that certain IL-2Rβ ligands of the IL-2Rβ ligand group having the amino acid sequences of SEQ ID NOs: 1-163, 164-182, 578-808, and 1028-1043 - Competitively binds to the binding site with each of the other IL-2Rβ ligands within the group of 2Rβ ligands, wherein the IL-2Rβ ligand having the amino acid sequence of SEQ ID NO: 1044 competes with the group of IL-2Rβ ligands for binding to the binding site However, IL-2 can be defined as a binding site that does not compete with IL-2Rβ ligands for binding to the binding site.

Certain IL-2Rβ ligands of the IL-2Rβ ligand group (SEQ ID NOs: 1-163, 164-182, 578-808, and 1028-1043) have binding affinities (IC ₅₀ ) for IL-2Rβ subunits of less than 100 μM with no detectable binding affinity (IC ₅₀ ) to IL-2Rγc.

An IL-2Rγc ligand having the amino acid sequence of SEQ ID NO:224 does not compete with a group of IL-2Rβ ligands for binding to the binding site.

IL-2Rβ ligands (SEQ ID NOs: 1-163, 164-182, 578-808, and 1028-1043) are IL-2Rβ ligands having the amino acid sequences of SEQ ID NOs: 58, 83, 142, 169, 170, 1033, and 663. 2Rβ ligands may be included.

This IL-2R binding site of the IL-2Rβ ligand can be characterized using the competitive binding assay described in Example 20, for example.

Certain IL-2Rγc ligands provided by this disclosure exhibit binding affinities for IL-2Rγc subunits of less than 100 μM and binding affinities (IC ₅₀ ) for IL-2Rβ subunits of greater than 100 μM.

Certain families of IL-2Rγc ligands bind IL-2Rγc subunits competitively with certain other families of IL-2Rγc ligands and non-competitively with other families of IL-2Rγc ligands.

Specific binding sites for the IL-2Rγc subunit are SEQ ID NOs: 194-210, 904-913, 211-233, 914-920, 234-245, 246-254, 921-922, 265-267, and 932-940 competitively binds to each of the other IL-2Rγc ligands within the group of IL-2Rγc ligands, and the IL-2Rγc ligand having the amino acid sequence of SEQ ID NO: 948 is A group of IL-2Rγc ligands can be defined as a binding site that does not compete for binding to the binding site.

Specific IL-2Rγc ligands of the group of IL-2Rγc ligands (SEQ ID NOS: 194-210, 904-913, 211-233, 914-920, 234-245, 246-254, 921-922, 265-267, and 932 ˜940) has a binding affinity (IC ₅₀ ) for the IL-2Rγc subunit of less than 100 μM and a binding affinity (IC ₅₀ ) for the IL-2Rβ subunit of greater than 100 μM.

An IL-2Rβ ligand having the amino acid sequence of SEQ ID NO:58 does not compete with the group of IL-2Rγc ligands for binding to the binding site.

IL-2Rγc ligands can include IL-2Rγc ligands having the amino acid sequences of SEQ ID NOs: 198, 202, 224, 236, 248, and 266.

The IL-2R binding sites of these IL-2Rγc ligands can be characterized using the competitive binding assays described in Example 20, for example.

Compounds provided by this disclosure, including IL-2Rβ ligands having the amino acid sequence of SEQ ID NOs: 1-163, 164-182, 578-80, or 1028-1043, bind to specific binding sites on IL-2Rβ subunits. IL capable of binding and having the amino acid sequence of SEQ ID NOs: 194-210, 904-913, 211-233, 914-920, 234-245, 246-254, 921-922, 265-267, or 932-940 Compounds provided by this disclosure, including -2Rγc ligands, can bind to specific binding sites on the IL-2Rγc subunit.

Methods of treating a disease in a patient provided by the present disclosure include administering to the patient a therapeutically effective amount of an IL-2Rβ ligand having the amino acid sequence of SEQ ID NOs: 1-163, 164-182, 578-80, or 1028-1043. , and/or an IL-2Rγc having the amino acid sequence of SEQ ID NOS: 194-210, 904-913, 211-233, 914-920, 234-245, 246-254, 921-922, 265-267, or 932-940 It can include administering a compound that includes a ligand.

A method of treating a disease in a patient provided by the present disclosure comprises administering to the patient a therapeutically effective amount of a compound that binds to the specific binding site of IL-2Rβ and/or the specific binding site of the IL-2Rγc subunit. can contain.

Methods provided by the present disclosure modulate the activity of IL-2R by providing a ligand or compound that interacts with a specific binding site of IL-2Rβ and/or a specific binding site of IL-2Rγc subunit. including how to

A method of diagnosing a disease in a patient provided by the present disclosure comprises administering to the patient an effective amount of a ligand or compound that binds to a specific binding site of IL-2Rβ and/or a specific binding site of IL-2Rγc subunit Including.

Peptides provided by the present disclosure can be synthesized by methods known in the art, for example, by using standard solid phase techniques. Standard methods include exclusive solid phase synthesis, partial solid phase synthesis methods, fragment condensation, classical solution synthesis, as well as recombinant DNA techniques. On solid phase, synthesis can be initiated from the C-terminus of the peptide using an α-amino protected resin. Suitable starting materials can be prepared, for example, by coupling the requisite α-amino acid to chloromethylated resin, hydroxymethyl resin, or benzhydrylamine resin. Peptides provided by the present disclosure can be prepared by coupling an α-amino protected amino acid to a chloromethylated resin using, for example, a cesium bicarbonate catalyst. After initial coupling, the α-amino protecting group can be removed by a choice of reagents including trifluoroacetic acid (TFA) or hydrochloric acid (HCl) solutions in organic solvents at room temperature.

α-Amino protecting groups are those known to be useful in the art of stepwise synthesis of peptides. Acyl-type protecting groups such as formyl, trifluoroacetyl, and acetyl, aromatic urethane-type protecting groups such as benzyloxycarbonyl (Cbz) and substituted Cbz, tert-butyloxycarbonyl (Boc), isopropyloxycarbonyl, and cyclohexyloxycarbonyl and alkyl-type protecting groups such as benzyl and triphenylmethyl. Boc and Fmoc are preferred protecting groups. Side chain protecting groups remain intact during coupling and are not cleaved during deprotection of the amino terminal protecting group or during coupling. Side chain protecting groups must be removable under reaction conditions that do not alter the target peptide once synthesis of the final peptide is complete.

Suitable side chain protecting groups for Tyr include tetrahydropyranyl, tert-butyl, trityl, benzyl, Cbz, Z-Br-Cbz, and 2,5-dichlorobenzyl. Suitable side chain protecting groups for Asp include benzyl, 2,6-dichlorobenzyl, methyl, ethyl, and cyclohexyl. Suitable side chain protecting groups for Thr and Ser include acetyl, benzoyl, trityl, tetrahydropyranyl, benzyl, 2,6-dichlorobenzyl, and Cbz. The side chain protecting group for Thr and Ser is benzyl. Suitable side chain protecting groups for Arg include nitro, tosyl (Tos), Cbz, adamantyloxycarbonylmesitoylsulfonyl (Mts), or Boc. Suitable side chain protecting groups for Lys include Cbz, 2-chlorobenzyloxycarbonyl (2-ClCbz), 2-bromobenzyloxycarbonyl (2-BrCbz), Tos, or Boc.

After removal of the α-amino protecting group, the remaining protected amino acids can be attached stepwise in the desired order. An excess of each protected amino acid is generally used in, for example, methylene chloride (CH ₂ Cl ₂ ), dimethylformamide (DMF) mixtures with a suitable carboxyl group activating agent such as dicyclohexylcarbodiimide (DCC) in solution. .

After the desired amino acid sequence is completed, the desired peptide is treated with reagents such as trifluoroacetic acid or hydrogen fluoride (HF), which not only cleaves the peptide from the resin, but also cleaves remaining side chain protecting groups. can be separated from the resin support by Hydrogen fluoride treatment results in the formation of the free peptide acid when chloromethylated resins are used. When using a benzhydrylamine resin, hydrogen fluoride treatment gives the free peptide amide directly. Alternatively, when using a chloromethylated resin, side-chain protected peptides can be treated with ammonia to give the desired side-chain protected amides, or with alkylamines to give side-chain protected alkylamides or dialkylamides, on the peptide resin. can be separated by processing Side chain protection is then removed in the usual manner by treatment with hydrogen fluoride to give the free amide, alkylamide, or dialkylamide. These solid-phase peptide synthesis procedures are well known in the art and can be used to determine the minimum size of peptides with such activity, as well as one, two, or more residues. It is also possible to generate all peptides forming a group of peptides that differ in group from the preferred motif (or the minimum size of that motif). Individual peptides can then be screened for their ability to bind to the IL-2Rβ and/or IL-2Rγc subunits. Peptide synthesis methods can also be used to synthesize truncated and deleted analogs and combinations of truncated and deleted analogs of all peptide compounds provided by this disclosure.

These procedures also apply to peptides in which amino acids other than the 20 naturally occurring genetically encoded amino acids are substituted at one, two, or more positions of any of the compounds of the invention. can also be used to synthesize For example, naphthylalanine can be replaced with tryptophan to facilitate synthesis. Other synthetic amino acids that can be substituted for the peptides of the present invention include d-amino acids such as L-hydroxypropyl, L-3,4-dihydroxyphenylalanyl, Ld-hydroxylysyl and Dd-methylalanyl, L- Included are a-methylalanyl, b-amino acids, and isoquinolyl. D-amino acids and non-naturally occurring synthetic amino acids can also be incorporated into the peptides provided by this disclosure.

The naturally-occurring side chains of the 20 genetically encoded amino acids (or D-amino acids) are complemented by other side chains such as alkyl, lower alkyl, cyclic 4-, 5-, 6-, to 7-membered alkyl, amido , amido-lower-alkyl, amido-di(lower-alkyl), lower-alkoxy, hydroxy, carboxy, and lower ester derivatives thereof, and 4-, 5-, 6-, to 7-membered heterocyclics, and the like. For example, proline analogs can be used in which the ring size of the proline residue varies from 5-, 4-, 6-, or 7-membered. Cyclic groups can be saturated or unsaturated, and if unsaturated, can be aromatic or non-aromatic.

Cyclic groups can be saturated or unsaturated, and if unsaturated, can be aromatic or non-aromatic. A heterocyclic group, for example, can contain one or more nitrogen, oxygen, and/or sulfur heteroatoms. Examples of such groups include furazanyl, furyl, imidazolidinyl, imidazolyl, imidazolinyl, isothiazolyl, isoxazolyl, morpholinyl (eg, morpholino), oxazolyl, piperazinyl (eg, 1-piperazinyl), piperidyl (eg, 1-piperidyl, and piperidino), pyranyl, pyrazinyl, pyrazolidinyl, pyrazolinyl, pyrazolyl, pyridazinyl, pyridyl, pyrimidinyl, pyrrolidinyl (e.g., 1-pyrrolidinyl), pyrrolinyl, pyrrolyl, thiadiazolyl, thiazolyl, thienyl, thiomorpholinyl (e.g., thiomorpholino), and triazolyl. be These heterocyclic groups may be substituted or unsubstituted. If the group is substituted, the substituent can be alkyl, alkoxy, halogen, oxygen, or substituted or unsubstituted phenyl.

Peptides provided by this disclosure can be modified, for example, by phosphorylation and by other methods known in the art. Thus, the peptides of this disclosure can also serve as a basis for preparing peptidomimetics with similar biological activity.

A variety of techniques for constructing peptidomimetics that have the same or similar desired biological activity as the corresponding peptide, but that have advantageous activities over the peptide with respect to solubility, stability, and susceptibility to hydrolysis and proteolysis. is available. The following describes methods for preparing peptidomimetics modified at the N-terminal amino group, C-terminal carboxyl group, and/or changing one or more of the amide bonds within the peptide to non-amide bonds. Two or more such modifications can be combined in one peptidomimetic structure (eg, modification at the C-terminal carboxyl group and inclusion of a —CH ₂ —carbamate bond between two amino acids within the peptide).

Peptides can be synthesized as free products or can be prepared as the corresponding amides or esters. The amino-terminus and/or carboxy-terminus of the peptide can also be modified. Amino terminal modifications include methylation (i.e., -NHCH ₃ or -NH(CH ₃ ) ₂ ), acetylation, addition of a carbobenzoyl group, or any blocking containing a carboxylate function defined by RCOO-. and blocking the amino terminus with a group, wherein R may be selected from the group consisting of naphthyl, acridinyl, steroidyl, and similar groups. Carboxy terminal modifications include replacing the free acid with a carboxamide group or forming a cyclic lactam at the carboxy terminus to introduce structural constraints. Amino terminal modifications also include, for example, alkylation, acetylation, addition of carbobenzoyl groups, and formation of succinimide groups.

In the preparation of peptidomimetics in which the C-terminal carboxyl group is replaced with an ester such as -C(O)OR, the side-chain protected peptide is treated with a base and a suitable alcohol such as methanol using a resin used in the preparation of peptide acids. disconnect. The side chain protecting groups can then be removed in the usual manner by treatment with hydrogen fluoride to give the desired ester.

Benzhydrylamine resin is used as a solid support for peptide synthesis in the preparation of peptidomimetics in which the C-terminal carboxyl group is substituted with amide-C(O)NR ³ R ⁴ . Once the synthesis is complete, hydrogen fluoride treatment to release the peptide from the support directly yields the free peptide amide (ie the C-terminus is -C(O) _NH2 ). Alternatively, use of a chloromethylating resin during peptide synthesis in combination with reaction with ammonia to cleave the side chain protected peptide from the support yields the free peptide amide, reaction with an alkyl or dialkylamine yields Side chain protected alkylamides or dialkylamides are obtained (ie the C-terminus is -C(O)NRR ¹ , where R and R ¹ are as defined above). Side chain protection can then be removed in the usual manner by treatment with hydrogen fluoride to give the free amide, alkylamide, or dialkylamide.

The C-terminal carboxyl group or C-terminal ester can be induced to cyclize by internal substitution of the —OH or ester (—OR) of the carboxyl group or ester with the N-terminal amino group, respectively, to form a cyclic peptide. . For example, after synthesis and cleavage to give the peptide acid, the free acid can be added to a suitable carboxyl group such as dicyclohexylcarbodiimide (DCC) in solution, for example methylene chloride ( _CH2Cl2 ), dimethylformamide (DMF) _mixtures . It is converted to an activated ester by an activating agent. A cyclic peptide can then be formed by internal displacement of the activated ester with the N-terminal amine.

Peptides can be cyclized so that there are no terminal amino or carboxyl groups, or desamino or a descarboxy residue can be incorporated. C-terminal functional groups of peptides include, for example, amido, amido-lower-alkyl, amido-di(lower-alkyl), lower-alkoxy, hydroxy, and carboxy, and lower ester derivatives thereof, and pharmaceutically acceptable salts thereof. be done.

The peptide compounds provided by this disclosure can also serve as structural models for non-peptide compounds with similar biological activity. One skilled in the art will construct compounds that have the same or similar desired biological activity as a particular peptide compound, but have preferred activity over the peptide compound with respect to, for example, solubility, stability, and susceptibility to hydrolysis and proteolysis. Recognize that a variety of techniques are available for These techniques involve replacing the peptide backbone with backbones composed of phosphonates, amidates, carbamates, sulfonamides, secondary amines, and N-methylamino acids.

Peptidyl mimetics with one or more of the peptidyl linkages -C(O)NH- include -CH ₂ -carbamate linkages, phosphonate linkages, -CH ₂ -sulfonamide linkages, urea linkages, secondary amine (-CH ₂ NH—) bond, or a bond as an alkylated peptidyl bond —C(O)NR ⁶ —, where R ⁶ is C _1-6 alkyl, and can be replaced at appropriate points during the synthesis. They can be prepared during conventional peptide synthesis by substituting appropriately protected amino acid analogues for amino acid reagents.

Suitable reagents include, for example, amino acid analogues in which the carboxyl group of the amino acid is replaced with a suitable moiety to form one of the above bonds. For example, if a -C(O)NR- in a peptide can be replaced by a _-CH2 -carbamate bond ( _-CH2OC (O)NR-), then the carboxyl (-COOH) group of a suitably protected amino acid is , is first reduced to a —CH ₂ OH group and then converted to —OC(O)Cl functional group or paranitrocarbonate —OC(O)O—C ₆ H ₄ —p—NO ₂ functional group by conventional methods. . Reaction of any such functional group with a free or alkylated amine on the N-terminus of a partially engineered peptide found on a solid support results in a —CH ₂ OC(O)NR— linkage. bring formation. Similarly, amide bonds in peptides can be replaced with phosphonate bonds.

Replacement of an amide bond in a peptide with a —CH ₂ -sulfonamide bond can be achieved by reducing the carboxyl (—COOH) group of a suitably protected amino acid to a —CH ₂ OH group, followed by The hydroxyl group is converted to a suitable leaving group such as a tosyl group by conventional methods. Reaction of the tosylated derivative with, for example, thioacetic acid, followed by hydrolysis and oxidative chlorination, replaces the carboxyl group of an otherwise appropriately protected amino acid --CH ₂ --S(O) ₂ Cl functionality. provide a base. Use of this appropriately protected amino acid analog in peptide synthesis provides for inclusion of a -CH ₂ S(O) ₂ NR- bond which replaces an amide bond within the peptide, thereby providing a peptidomimetic. Replacement of amide bonds within peptides with urea bonds can be accomplished using similar methods.

A secondary amine bond, where the —CH ₂ NH— bond replaces the amide bond in the peptide, can be used, for example, by using an appropriately protected dipeptide analog in which the carbonyl bond of the amide bond has been reduced to a CH ₂ group by conventional methods. can be prepared by For example, in the case of diglycine, reduction of the amide to the amine is followed by _{deprotection H2NCH2CH2NHCH2COOH} , which is then _used in the N-protected _form in the next coupling reaction. Preparation of such analogs by reduction of the carbonyl group of the amide bond within the dipeptide is known in the art.

Suitably protected amino acid analogs can be used in conventional peptide synthesis in a manner analogous to the corresponding amino acids. For example, typically about 3 equivalents of protected amino acid analog can be used in this reaction. When an inert organic diluent such as methylene chloride or DMF is used and acid is produced as a reaction by-product, the reaction solvent is typically used in excess to remove the acid produced during the reaction. Contains tertiary amines. One particularly preferred tertiary amine is diisopropylethylamine, typically used in about a 10-fold excess. The reaction results in the incorporation of amino acid analogs with non-peptidyl bonds into the peptidomimetic. Such substitutions can be repeated as desired such that zero to all amide bonds within the peptide are replaced with non-amide bonds.

Peptides provided by the present disclosure can be cyclized so that there are no terminal amino or carboxyl groups to reduce susceptibility to proteases or restrict the conformation of the peptide, or the peptide can incorporate a desamine or decarboxy residue at the end of the The C-terminal functional groups of the compounds of the present invention include amido, amido-lower alkyl, amido-di(lower alkyl), lower alkoxy, hydroxy, and carboxy, and lower ester derivatives thereof and pharmaceutically acceptable salts thereof. included.

Peptides provided by the present disclosure can exist in cyclized forms with intramolecular disulfide bonds between cysteine thiol groups. Alternatively, an intermolecular disulfide bond between the cysteine thiol groups can be generated, resulting in a dimeric (or higher oligomeric) compound. One or more of the cysteine residues may be replaced with homocysteine. One of the sulfurs can be replaced by a _CH2 group or other isostere in place of the sulfur. These analogs can be made through intramolecular or intermolecular substitutions. Those skilled in the art will readily appreciate that this substitution can also occur using the a-amino-g-butyric acid derivatives shown above and other homologues of homocysteine.

Alternatively, the amino terminus of the peptide can be capped with an α-substituted acetic acid, the α-substituent leaving an α-haloacetic acid such as α-chloroacetic acid, α-bromoacetic acid, or α-iodoacetic acid. is the base. Peptides provided by the present disclosure can be cyclized or dimerized through replacement of the leaving group by a sulfur of a cysteine or homocysteine residue.

Compounds comprising IL-2R agonists and IL-2Rβ ligands and/or IL-2Rγc ligands can be, for example, synthetic peptides, conjugates of peptides to another peptide or protein, recombinant fusion proteins, or single chain peptides. could be.

An IL-2R agonist compound can be an IL-2Rβ ligand provided by the disclosure, an IL-2Rγc ligand provided by the disclosure, or an IL-2Rβ ligand provided by the disclosure and an IL-2Rγc ligand provided by the disclosure. Both ligands can be included.

IL-2R agonist compounds are IL-2Rβ ligands provided by this disclosure, such as the IL-2Rβ ligands of SEQ ID NOs: 1-193, 578-903, or 1028-1050, the IL-2Rβ ligands of SEQ ID NOs: 194-267 or 904-1027. IL-2Rγc ligands provided by the present disclosure, such as -2Rγc ligands, or both IL-2Rβ ligands provided by the present disclosure and IL-2Rγc ligands provided by the present disclosure can be included.

Peptides provided by the present disclosure can comprise an IL-2Rβ ligand or an IL-2Rγc ligand and additional amino acids.

The additional amino acids are at the C-terminus of the IL-2Rβ or IL-2Rγc ligand, the N-terminus of the IL-2Rβ or IL-2Rγc ligand, or both the C-terminus and the N-terminus of the IL-2Rβ or IL-2Rγc ligand. can be combined.

A peptide comprising an IL-2Rβ ligand and/or an IL-2Rγc ligand can contain, for example, 10-50 amino acids, 10-40 amino acids, 10-30 amino acids, or 15-25 amino acids.

A peptide comprising an IL-2Rβ ligand and/or an IL-2Rγc ligand can contain, for example, 5-300 amino acids, 10-200 amino acids, or 10-100 amino acids.

Peptides comprising an IL-2Rβ ligand and an IL-2Rγc ligand can have at least substantially the same binding affinity (IC ₅₀ ) for their respective human IL-2 subunits as their respective IL-2R ligands alone. can.

A compound comprising an IL-2Rβ ligand and/or an IL-2Rγc ligand can be a conjugate.

A conjugate can comprise one or more IL-2Rβ ligands, one or more IL-2Rγc ligands, or a combination thereof.

A conjugate can be a homodimer comprising two IL-2Rβ ligands or two IL-2Rγc ligands.

A conjugate can be a heterodimer comprising at least one IL-2Rβ ligand and at least one IL-2Rγc ligand.

A conjugate can be a heterodimer comprising at least one IL-2Rβ ligand and at least one IL-2Rγc ligand covalently attached to a protein.

A conjugate can include a linker, which is configured to link an IL-2Rβ ligand and/or IL-2Rγc ligand to one or more other IL-2Rβ and/or IL-2Rγc ligands. The linker can be attached to the IL-2Rβ and/or IL-2Rγc ligand and any additional moieties by covalent bonds or by non-covalent bonds such as ionic bonds.

Each IL-2Rβ ligand and/or IL-2Rγc ligand can be independently attached to a linker via the C-terminus, the N-terminus, or both the C-terminus and the N-terminus.

For example, in a homodimer, the C-terminus of a first IL-2Rβ ligand and the C-terminus of a second IL-2Rβ ligand can be attached to a linker, the N-terminus of the first IL-2Rβ ligand and The N-terminus of the second IL-2Rβ ligand can be attached to the linker, or the C-terminus of the first IL-2Rβ ligand and the N-terminus of the second IL-2Rβ ligand can be attached to the linker. can.

For example, in a homodimer, the C-terminus of a first IL-2Rγc ligand and the C-terminus of a second IL-2Rγc ligand can be attached to a linker, the N-terminus of the first IL-2Rγc ligand and The N-terminus of the second IL-2Rγc ligand can be attached to the linker, or the C-terminus of the first IL-2Rγc ligand and the N-terminus of the second IL-2Rγc ligand can be attached to the linker. can.

For example, in the heterodimer, the C-terminus of the IL-2Rβ ligand and the C-terminus of the IL-2Rγc ligand can be attached to a linker, and the N-terminus of the IL-2Rβ ligand and the N-terminus of the IL-2Rγc ligand are The C-terminus of the IL-2Rβ ligand and the N-terminus of the IL-2Rγc ligand can be conjugated to a linker, or the N-terminus of the IL-2Rβ ligand and the C-terminus of the IL-2Rγc ligand can be conjugated to , can be attached to the linker.

A heterodimeric compound comprising an IL-2Rβ ligand and an IL-2Rγc ligand can be configured to activate the IL-2 receptor.

A heterodimeric compound comprising an IL-2Rβ ligand and an IL-2Rγc ligand can be configured to activate the IL-2 receptor without activating cells expressing the IL-2Rα ligand.

For example, primary human peripheral blood mononuclear cells (PBMC) expressing IL-2Rβγc subunits phosphorylate transcription 5 (STAT5) when incubated with a heterodimeric compound containing an IL-2Rβ ligand and an IL-2Rγc ligand. Primary human peripheral blood mononuclear cells (PBMC) that oxidize and express the IL-2Rα (CD25) subunit do not phosphorylate transcription 5 (STAT5).

The heterodimer can comprise an IL-2Rβ ligand, an IL-2Rγc ligand, and a linker configured such that the heterodimer is an agonist of the IL-2 receptor.

The linker can include lengths that facilitate binding of IL-2Rβ and/or IL-2Rγc ligands to the IL-2 receptor. For example, linkers can have a length, eg, from 20 Å to 100 Å, 30 Å to 80 Å, or 40 Å to 60 Å.

The linker can include chemical structures that facilitate binding of IL-2Rβ and/or IL-2Rγc ligands to the IL-2 receptor. For example, linkers can include peptides, or carbohydrates.

Peptide linkers can include, for example, 5-100 amino acids, 5-80 amino acids, 5-60 amino acids, 5-40 amino acids, 5-20 amino acids, or 5-10 amino acids. Peptide linkers can include, for example, 2-100 amino acids, 2-80 amino acids, 2-60 amino acids, 2-40 amino acids, 2-20 amino acids, 5-10 amino acids, or 2-5 amino acids.

A hydrocarbon linker can be polyethylene oxide. Polyethylene oxide can have a structure of the formula -(O-(CH ₂ ) ₂ -) _n -(CH ₂ ) ₂ -, where n is an integer from 1 to 30.

Hydrocarbon linkers can be derived from polyethylene oxide having a structure of the formula H ₂ N—(O—(CH ₂ ) ₂ —) _n —(CH ₂ ) ₂ —COOH, where n is an integer from 1 to 30. .

A peptide dimeric compound can comprise two monomeric subunits, wherein the peptide dimeric compound comprises an IL-2Rβ ligand and/or an IL-2Rγc ligand. The monomeric subunits present in the peptide dimeric compound can be linked at either their C- or N-termini or via internal amino acid residues such as linker moieties. Both monomeric subunits can be linked via their respective N-termini, both monomeric subunits can be linked via their respective C-termini, or both monomeric subunits can be linked via their respective C-termini Subunits can be linked through internal amino acid residues. One monomeric subunit is linked to another monomeric subunit either through its N-terminus, C-terminus, or by an internal amino acid. and the linkage can occur through the same or different amino acid residues on the two monomeric subunits of the peptide dimeric compound. The monomeric subunits of a peptide dimeric compound can be linked via both their N- and C-termini. The two N-termini of the monomeric subunits can be linked, the two C-termini of the monomeric subunits can be linked, and the N-terminus of the first monomeric subunit can be linked to the peptide dimer. can be linked to the C-terminus of the second monomeric subunit of the compound, the C-terminus of the first monomeric subunit to the N-terminus of the second monomeric subunit of the peptide dimeric compound; can be concatenated.

Linkers can comprise peptides or non-peptides. A linker portion can comprise any suitable structure, length, and/or size. Linker moieties are, for example, DIG, PEG13, PEG25, PEG1K, PEG2K, PEG3.4K, PEG4K, PEG5K, IDA, IDA-Palm, IDA-Boc, IDA-Ac, IDA- isovaleric acid, ADA triazine, triazine-Boc, isophthalic acid, 1,3-phenylenediacetic acid, Glu, Asp, D-Glu, D-Asp, 1,4-phenylenediacetic acid, biphenyldiacetic acid, cyclopropylacetic acid, Succinic acid, glutaric acid, dodecanedioic acid, suitable aliphatic diacids, suitable aromatic diacids, heteroaromatic compounds, and polyethylene glycols having molecular weights of, for example, 400 Da to 40,000 Da can be included. When the linker is IDA, ADA, or any linker with a free amine, 2-me-trifluorobutyl, trifluoropentyl, acetyl, octonyl, butyl, pentyl, hexyl, palmityl, lauryl, oleoyl, lauryl, tri fluoromethylbutyric acid, cyclopentanecarboxylic acid, cyclopropylacetic acid, 4-fluorobenzoic acid, 4-fluorophenylacetic acid, 3-phenylpropionic acid, tetrahydro-2H-pyran-4-carboxylic acid, succinic acid, and glutaric acid, 10 It can be acylated with acylating organic compounds such as linear fatty acids with ˜20 carbon units, cholic acid, and other bile acids. Small PEGs (PEG4-PEG13), Glu, IsoGlu, or Asp can be used as spacers prior to acylation.

A linker can connect two monomeric subunits by connecting two sulfur containing C-terminal or N-terminal amino acids. Two sulfur-containing amino acids can be connected by a linker comprising a dihalide, an aliphatic chain, or PEG. A linker can connect two monomeric subunits by linking a sulfur containing C-terminal amino acid to the C-terminus of each monomeric subunit. A linker can connect two monomeric subunits by linking the sulfur containing N-terminal amino acids to the N-terminus of each monomeric subunit. A linker can connect two monomeric subunits by connecting the sulfur-containing C-terminal amino acid of one monomeric subunit to the sulfur-containing N-terminal amino acid of the other monomeric subunit. Two sulfur-containing amino acids are homobifunctional maleimide crosslinkers, dihalides, 1,2-bis(bromomethyl)benzene, 1,2-bis(chloromethyl)benzene, 1,3-bis(bromomethyl)benzene, 1, 3-bis(chloromethyl)benzene, 1,4-bis(bromomethyl)benzene, 1,4-bis(chloromethyl)benzene, 3,3'-bis-bromomethyl-biphenyl, or 2,2'-bis-bromomethyl -biphenyl-containing linkers. Examples of haloacetyl crosslinkers contain iodoacetyl or bromoacetyl groups. These homobifunctional linkers may include spacers containing PEG or aliphatic chains. Linkers can be bifunctional linkers (e.g., diacids, diamines, dihalides, N-hydroxysuccinamine (NHS) activated diester, bismaleimide).

Examples of suitable linkers include Zig, PEG4, PEG4-Biotin, PEG13, PEG25, PEG1K, PEG2K, PEG3.4K, PEG4K, PEG5K, IDA, ADA, Boc-IDA, glutaric acid, isophthalic acid, 1,3- phenylenediacetic acid, 1,4-phenylenediacetic acid, 1,2-phenylenediacetic acid, triazines, Boc-triazines, IDA-biotin, PEG4-biotin, AADA, aliphatic groups, aromatic groups, heteroaromatic groups, and Polyethylene glycol-based linkers with molecular weights from 400 Da to 40,000 Da are included. Examples of suitable bifunctional linkers include bifunctional linkers (e.g., , diacids, diamines, dihalides, N-hydroxysuccinamine (NHS) activated diesters, bismaleimides).

Peptide monomers and compounds can form cyclized structures through disulfide, lactam, olefinic, triazole, selenoether or diselenide bonds. The cyclized structure of each peptide ligand can, in some cases, increase the potency and selectivity of ligands and compounds containing ligands.

For example, heterodimers are IL-2Rβ ligands provided by this disclosure, such as the IL-2Rβ ligands of SEQ ID NOs: 1-193, 578-903, or 1028-1050, SEQ ID NOs: 194-267 and 904-1027. IL-2Rγc ligands provided by the present disclosure, such as IL-2Rγc ligands, and linkers such as polyethylene glycol-based linkers having a length of 10 Å to 60 Å can be included.

Individual IL-2Rβ and IL-2Rγc ligands can be linked in a variety of ways to produce heterodimers that can be evaluated for IL-2R agonistic, antagonistic, or other related activities. Agonist activity may depend on the heterodimer binding simultaneously to both IL-2Rβ and IL-2Rγc subunits to induce proximity and orientation compatible with signal transduction. Several compound characteristics can affect heterodimer activity such as, for example, monomer peptide linker structure, linker length, peptide ligand orientation, and ECD binding site specificity.

Since IL-2 receptor signaling can be compatible with a range of induced subunit orientations, linkers can facilitate binding outside the IL-2R binding site(s). The dimensions of the quaternary complex of IL-2 with the IL-2Rα, IL-2Rβ, and IL-2Rγc subunits are that linkers with lengths up to about 50 Å connect the IL-2Rβ and IL-2Rγc ligands. , may be useful for inducing productive subunit alignment and agonist activity.

For example, a heterodimer comprising an IL-2Rβ ligand and an IL-2Rγc ligand can be attached via a PEG linker. An alkyne functional group can be attached to the distal end of the PEG linker of the peptide. The other peptide monomer can be functionalized with a terminal azide. Monomers are converted to triazoles under conditions compatible with maintenance of disulfide bridges present in certain peptide ligands using, for example, a 1,3 dipolar Huisgen cycloaddition reaction between an azide and an alkyne. It can be conjugated using click chemistry to form a bond. Alkyne and azide groups can be incorporated into each monomer using commercially available amino acid building blocks. The spacing between the two peptide ligands can be selected using PEG-linkers of various lengths. Total linker lengths from 10 Å or less to 50 Å or more can be synthesized using commercially available Fmoc-PEG amino acids, providing a wide range of heterodimeric linker length diversity.

The induced receptor subunit orientation and the likelihood of proper intracellular alignment and signaling may be, in part, a function of the orientation in which the peptide ligands ligate to form heterodimers. To determine the preferred induced subunit orientation, the peptide ligands are attached via a suitable linker at the C-terminus of both subunit-binding ligands and at the N-terminus of both subunit-binding ligands. linked via a linker, or the N-terminus of one linked subunit linked through the C-terminus of the other linked subunit via a suitable linker in any of four possible orientations. (two scenarios are possible). Heterodimers can also be linked through amino acid side chains. Heterodimers can also be linked through amino acid side chains. The heterodimer linkage orientation can be engineered, for example, by synthesizing ligand monomers with click functionalities, i.e., azide or alkyne, and a PEG-linker at either the N- or C-terminus. .

Peptides provided by the present disclosure may comprise at least one IL-2Rβ ligand and/or at least one IL-2Rγc ligand. Peptides can include, for example, less than 50 amino acids, which can include amino acids that constitute at least one IL-2Rβ ligand and/or at least one IL-2Rγc ligand.

Peptides comprising IL-2Rβ and/or IL-2Rγc ligands contain, for example, 5-100 amino acids, 5-80 amino acids, 5-50 amino acids, 10-40 amino acids, 10-30 amino acids, or 15-25 amino acids. be able to.

In addition to IL-2Rβ and/or IL-2Rγc ligands, peptides may be used, for example, to establish the conformation of IL-2Rβ and/or IL-2Rγc ligands, or IL-2Rβ and/or IL-2Rγc ligands. Additional amino acids can be included for linking the ligand to other compounds. Additional amino acids can be attached to the N-terminus and/or C-terminus of the IL-2Rβ ligand and/or IL-2Rγc ligand.

Conjugates provided by the present disclosure comprise at least one IL-2Rβ ligand and/or at least one IL-2Rγc ligand.

A conjugate can include a polypeptide.

A polypeptide can be a single chain peptide with more than one IL-2Rβ ligand and/or more than one IL-2Rγc ligand. IL-2Rβ ligand and/or IL-2Rγc ligand can be attached through an amino acid linker.

Amino acid linkers can include, for example, more than 1 amino acid, more than 5 amino acids, more than 10 amino acids, more than 50 amino acids, or more than 100 amino acids. Peptide linkers can comprise 1-100 amino acids, eg, 3-75 amino acids, 5-50 amino acids, or 10-25 amino acids.

For example, in homodimers, can the C-terminus of a first IL-2Rβ ligand and/or IL-2Rγc ligand and the C-terminus of a second IL-2Rβ ligand and/or IL-2Rγc ligand be attached to a linker? , the N-terminus of the first IL-2Rβ ligand and/or IL-2Rγc ligand and the N-terminus of the second IL-2Rβ ligand and/or IL-2Rγc ligand may be attached to a linker, or the first IL- The C-terminus of a 2Rβ ligand and/or IL-2Rγc ligand and the N-terminus of a second IL-2Rβ ligand and/or IL-2Rγc ligand can be attached to a linker.

Polypeptides comprising IL-2Rβ ligands and/or IL-2Rγc ligands provided by this disclosure can be, for example, 5 amino acids to 4,000 amino acids, 5 amino acids to 3,000 amino acids, 5 amino acids to 2,500 amino acids, or It can contain from 5 amino acids to 2,000 amino acids.

Polypeptides can be synthetic or recombinant polypeptides.

The single-chain peptide comprises at least one IL-2Rβ ligand and/or IL-2Rγc ligand, one or more IL-2Rα ligands, one or more IL-2Rβ ligands, and/or one or more IL-2Rγc ligands. It can be a heteromer having a combination of ligands. For example, a single peptide chain can include an IL-2Rα ligand, an IL-2Rβ ligand, and/or an IL-2Rγc ligand with an amino acid linker that couples adjacent ligands. Single-chain peptides can further comprise additional amino acids at the N-terminus and/or C-terminus of the polypeptide.

IL-2Rα, IL-2Rβ, and/or IL-2Rγc ligands can be arranged in any order.

Each of the adjacent ligands is connected via the N-terminus of each ligand, via the C-terminus of each ligand, via the N-terminus and C-terminus of adjacent ligands, or via side chains of ligands and/or linkers. Can be combined independently.

For example, in a heteromer, the C-terminus of an IL-2Rα ligand can be attached to a linker, the N-terminus of an IL-2Rβ ligand or the N-terminus of an IL-2Rγc ligand can be attached to a linker, and the IL-2Rβ ligand The C-terminus or N-terminus of an IL-2Rγc ligand can be attached to a linker, or the N-terminus of an IL-2Rα ligand can be attached to a linker.

Individual IL-2Rβ and/or IL-2Rγc ligands can be linked in a variety of ways to produce homodimers or homomers, heteromers that can be evaluated for IL-2R agonist and/or IL-2R antagonist activity. obtain. For example, homodimers of IL-2Rα ligands, or heterodimers of IL-2Rα ligands with IL-2Rβ ligands and/or IL-2Rγc can function as IL-2R antagonists. Agonist activity may depend on heteromers binding both IL-2Rβ and IL-2Rγc subunits simultaneously to induce proximity and orientation compatible with signaling or inhibition. Some compound characteristics, e.g., linker structure, linker length, peptide ligand orientation, ECD binding site specificity of monomeric peptides, and homodimer or It can affect the activity of the heteromer. IL-2R agonist and IL-2R antagonist activity can be measured, for example, by the affinity of IL-2Rα ligands for IL-2Rα subunits, the affinity of IL-2Rβ ligands for IL-2Rβ subunits, and/or the affinity of IL-2Rγc ligands for IL-2Rβ subunits. It may rely on increasing affinity for the IL-2Rγc subunit. The induced receptor subunit orientation and the likelihood of proper intracellular alignment and signaling may be, in part, a function of the orientation with which the peptide ligands are linked to form heteromers. To determine the preferred induced subunit orientation, adjacent ligands IL-2R are attached via a suitable linker to the C-terminus of both subunit-binding ligands and both subunit-binding ligands. or the N-terminus of one binding subunit through the C-terminus of the other binding subunit through a suitable linker in any of four possible orientations. can be combined with Homomers and heteromers can also be linked through amino acid side chains. Heteromeric linkage orientation can be engineered, for example, by synthesizing ligand monomers with a click functionality, ie, an azide or alkyne, and a PEG-linker at either the N- or C-terminus.

The polypeptide can be a synthetic modified polypeptide comprising one or more IL-2Rβ ligands and/or one or more IL-2Rγc ligands. Modifications can affect, for example, the activity of a polypeptide or the pharmacokinetics of a polypeptide. Examples include polypeptides incorporating polyethylene glycol moieties or albumin binding moieties.

Compounds containing an IL-2Rβ ligand, an IL-2Rγc ligand, or both an IL-2Rβ ligand and an IL-2Rγc ligand include fusion proteins.

The IL-2Rβ ligand and/or IL-2Rγc ligand can be fused to another protein that confers the desired functionality to the construct. For example, proteins can confer a desired pharmacokinetic profile or can be designed to target specific antigens.

Examples of suitable fusion partners include Fc fusion proteins, IgG fusion proteins, human serum albumin (HSA) fusion proteins, other human proteins and mutants and/or variants thereof and hydrophilic biodegradable protein polymers. is mentioned. Fusion protein partners can be naturally occurring proteins, modified naturally occurring proteins, or synthetic proteins.

For example, an IL-2Rβ ligand and/or an IL-2Rγc ligand provided by this disclosure can be fused to a protein that increases the circulating half-life of the compound. Fusion of a therapeutic protein to an IgG or IgG Fc domain increases the hydrodynamic radius of the protein, thus reducing renal clearance and the neonatal Fc receptor (FcRn)-mediated recycling of the fusion protein. It achieves this through, thus prolonging circulation half-life. Other fusion proteins can be designed to modify properties such as pharmacokinetics, biodistribution, pharmacodynamics, pharmacology, cytotoxicity, and/or targeting.

A fusion protein provided by the present disclosure can comprise a peptide provided by the present disclosure, or multiple tandem peptides, linked to one or more fusion protein partners. Fusion protein partners can be attached to the N-terminus and/or C-terminus of the tandem peptide. One or more fusion protein partners can be attached to the N-terminus and/or C-terminus of the tandem peptide. The IL-2Rβ ligand and/or IL-2Rγc ligand can be linked to one or more fusion protein partners, each fusion protein partner can be the same, or some of the fusion protein partners can be peptide can be different from others of the fusion protein partners linked to.

The amino acid sequence at the junction between the IL-2Rβ and/or IL-2Rγc ligand and the fusion partner protein can be either a direct fusion of the two protein sequences or a fusion with an intervening linker peptide. A linker peptide may be included as a spacer between the two protein moieties. Linker peptides can promote proper protein folding and stability of the component protein portions, improve protein expression, and enhance biological activity of the component protein portions. Peptide linkers used in fusion proteins can be designed to be unstructured flexible peptides. Peptide linkers are, for example, GS, GGS, GGGS, GGGGS, (GGGGGs)3, (Gly)8, (Gly)6, (EAAAK)1-3, A(EAAAK)4ALEA(EAAAK)4A, PAPAP, AEAAAAKEAAAKA, (Ala-Pro)a(10-34aa), disulfides, VSQTSKLTR AETVFPDV, PLG LWA, and GFLG, RVQDVIERFWDFIDQLSGSGSGK, and VDADGPLARLKKAIFSPGSGSGK, (PA)n (where n is an integer 1-20, such as (PA)10), and glycine- and serine-rich, such as repeating sequences such as (GS)n, where n is an integer 1-20, such as (GS)10. A flexible linker peptide with a fully extended β-strand conformation can have an end-to-end length of about 3.5 Å per residue. Thus, a 5-, 10-, 15-, or 10-residue linker peptide has a maximum overall length of 17.5 Å, 35 Å, 52.5 Å, 70 Å, 140 Å, or greater than 140 Å, respectively.

The linker peptide facilitates the engagement of the IL-2Rβ ligand and/or IL-2Rγc ligand with the IL-2Rβ subunit and/or IL-2Rγc subunit, and the IL-2Rβ ligand and/or IL-2Rγc ligand with IL facilitating proper conformation and orientation of the individual fusion protein moieties to facilitate binding to the -2 receptor, allow fusion protein recycling, and extend the circulating half-life of the active moiety; can be done. Because the factors that influence these interactions are difficult to predict, linker peptide requirements and appropriate lengths must be tested and determined empirically.

Multiple options exist for the design and construction of fusion proteins that can be selected to obtain molecules containing IL-2Rβ and/or IL-2Rγc ligands and having desired biological activity and pharmaceutical characteristics. Design options include, for example, the nature of the IL-2 selective agonist, the choice of partner protein portion, the organization of the fusion partner in the fusion protein, and the amino acid sequence at the junction between the IL-2R ligand and the fusion partner protein. mentioned.

In general, the preparation of fusion proteins provided by the present disclosure can be performed by, for example, polymerase chain amplification (PCR), preparation of plasmid DNA, digestion of DNA with restriction enzymes, preparation of oligonucleotides, ligation of DNA, isolation of mRNA, This can be accomplished by recognized recombinant DNA techniques, including introduction of the DNA into suitable cells, host transformation or transfection, and host culture. Additionally, fusion proteins can be isolated and purified using chaotropic agents and well-known electrophoretic, centrifugation, and chromatographic methods.

Genes encoding fusion proteins provided by this disclosure may include restriction enzyme digestion and ligation as the basic steps used to obtain the DNA encoding the desired fusion. The ends of the DNA fragments may require modification prior to ligation, which may include filling in overhangs, deleting the terminal portion of the fragment(s) with a nuclease (e.g. ExoIII), site-directed mutagenesis This can be achieved by introduction, or by adding new base pairs by PCR. Polylinkers and adapters can be used to facilitate joining of selected fragments. Expression constructs can be assembled step-by-step using rounds of restriction, ligation, and transformation in E. coli. Large numbers of cloning vectors suitable for construction of expression constructs are known in the art. The choice of cloning vector may be influenced by the gene transfer system chosen for introducing the expression construct into the host cell. At the end of each step, the resulting constructs may be analyzed by restriction, DNA sequencing, hybridization and PCR analysis.

Site-directed mutagenesis can be used to introduce specific mutations into genes encoding fusion proteins provided by this disclosure by methods known in the art. Any suitable site-directed mutagenesis procedure can be used in the present invention. There are many commercially available kits that can be used to prepare variants of the invention.

Various promoters (transcription initiation regulatory regions) may be used in accordance with the present invention. Selection of an appropriate promoter may depend on the proposed expression host. Promoters from heterologous sources may be used as long as they function in the host of choice.

Various signal sequences may be used to facilitate expression of the proteins described herein. A signal sequence can be selected or designed and used for efficient secretion and processing in the expression host. A signal sequence homologous to the human IL-2 coding sequence can be used for mammalian cells. Other suitable signal sequence/host cell pairs include B. B. subtilis for secretion in B. subtilis. subtilis sacB signal sequence and Saccharontyces cerevisiae α-mating factor, or P. P. pastoris secretion pastoris acid phosphatase phoI signal sequence. The signal sequence can be linked directly to the protein-coding sequence, through a sequence encoding a signal peptidase cleavage site, or through a short nucleotide bridge.

Elements for enhancing transcription and translation have been identified for eukaryotic protein expression systems. For example, flanking the cauliflower mosaic virus (CaMV) promoter 1000 bp to both sides of a heterologous promoter can increase transcription levels 10-400 fold in plant cells. The expression construct should also contain a suitable translation initiation sequence. Modification of expression constructs containing the Kozak consensus sequence for proper translation initiation can increase translation levels ten-fold.

The expression cassette is ligated into an appropriate vector compatible with the host being used. The vector must be able to accommodate the DNA sequence encoding the fusion protein to be expressed. Suitable host cells include eukaryotic and prokaryotic cells, such as cells that are easily transformed and exhibit rapid growth in culture medium. Examples of suitable host cells include E. Prokaryotes such as E. coli, Bacillus subtillus, as well as animal cells and yeast strains such as S. eukaryotes such as S. cerevisiae. Suitable mammalian cells include HEK, J558, NSO, SP2-O, or CHO. Other suitable hosts include, for example, insect cells such as Sf9. Conventional culture conditions can be used. Stable transformed or transfected cell lines can then be selected. In vitro transcription-translation systems can also be used as expression systems.

Nucleic acid encoding the desired fusion protein can be introduced into the host cell by standard techniques for transfecting cells. The term "transfecting" or "transfection" includes calcium phosphate co-precipitation, DEAE-dextran-mediated transfection, lipofection, electroporation, microinjection, viral transduction and/or integration of nucleic acids into host cells. It is intended to cover all conventional techniques for introduction.

Alternatively, synthetic genetic constructs can be used for all or part of the construction of the fusion proteins described herein. This may involve the in vitro synthesis of polynucleotide molecules designed to encode the polypeptide molecule of interest. Gene synthesis can be carried out using several techniques such as multiplex microchip-based techniques and similar techniques in which oligonucleotides are synthesized and assembled on an optically programmable microfluidic chip.

Fusion proteins provided by the present disclosure can be isolated from harvested host cells or from culture medium. Standard protein purification techniques are used to isolate the protein of interest from culture medium or from harvested cells. For example, purification techniques are used to express and purify the desired fusion protein at large scale (i.e., at least milligram quantities) from a variety of approaches including roller bottles, spinner flasks, tissue culture plates, bioreactors, or fermentors. can do.

Compounds provided by the present disclosure comprise at least one IL-2Rβ ligand and/or at least one IL-2Rγc ligand. The compound may be, for example, 1-10 IL-2Rβ and/or IL-2Rγc ligands, 1-6 IL-2Rβ and/or IL-2Rγc ligands, or 1-3 IL-2Rβ ligands and /or an IL-2Rγc ligand can be included. Examples of compounds comprising IL-2Rβ and/or IL-2Rγc ligands include peptides and conjugates. Examples of conjugates include one or more IL-2Rβ ligands and/or one or more IL-2Rγc bound to polypeptides, macromolecules such as polyethylene glycol, fusion proteins, or biological molecules such as antibodies. ligands.

Functionally, compounds comprising at least one IL-2Rβ ligand and/or at least one IL-2Rγc ligand are IL-2Rβγc agonists, IL-2Rαβγc agonists, IL-2Rβγc antagonists, IL-2Rαβγc antagonists, diagnostic reagents, imaging It can be a reagent, a targeting compound, a cytotoxic compound, and a compound that exhibits dual pharmacology.

Compounds comprising IL-2Rβ ligands and/or IL-2Rγc ligands provided by the present disclosure can be, for example, 1,000-400,000 Da, It can have a molecular weight of 000 Da to 20,000 Da, 1,500 Da to 15,000 Da, 2,000 Da to 10,000 Da, or 5,000 Da to 10,000 Da.

Compounds, including IL-2Rβ ligands and/or IL-2Rγc ligands provided by this disclosure, can be conjugated to one or more moieties that impart properties to the compounds that enhance therapeutic efficacy. Examples of properties include potency, water solubility, polarity, lipophilicity, pharmacokinetics, targeting, bioavailability, pH dependent binding, bioactivity, pharmacokinetics, cellular activity, metabolism, efficacy, non-reversible (caging). , selectivity, or a combination of any of the foregoing.

Compounds containing IL-2Rβ and/or IL-2Rγc ligands may contain one or more moieties that are cleavable in vivo. A moiety may be cleavable in a target-specific environment, eg, by a target-specific or target-enriching enzyme, or pH. Moieties may be cleavable upon exposure to electromagnetic energy, such as visible or infrared light, and/or by exposure to thermal energy.

Compounds containing IL-2Rβ and/or IL-2Rγc ligands can include polymers, peptides, or antibodies.

Compounds comprising IL-2Rβ ligands and/or IL-2Rγc ligands may include tumor targeting moieties such as tumor-specific antibodies, tumor-specific antibody fragments, tumor-specific proteins, tumor-specific peptides, non-peptidyl tumor cell ligands, or a combination of any of the foregoing.

Compounds comprising IL-2Rβ ligands and/or IL-2Rγc ligands can be administered with immune cell targeting moieties such as immune cell-specific antibodies, immune cell-specific antibody fragments, immune cell-specific proteins, immune cell-specific peptides, non- It can include peptidyl immune cell ligands, or combinations of any of the foregoing.

Compounds containing IL-2Rβ ligands and/or IL-2Rγc ligands can contain one or more caged molecules. Caged molecules can actually encapsulate compounds and serve to prevent biological activity in specific tissues, eg, protecting peripheral tissues from the toxicity of IL-2Rαβγc activation.

Compounds containing IL-2Rβ and/or IL-2Rγc ligands can include moieties, which include small molecules, peptides, polymers, or antibodies. Small molecules can be non-peptidyl molecules. Moieties can exhibit pharmacological effects. A pharmacological effect is exhibited upon binding of the moiety to an IL-2Rβ ligand and/or IL-2Rγc ligand and/or after cleavage of the moiety from a compound containing the IL-2Rβ ligand and/or IL-2Rγc ligand. can be done.

A compound comprising an IL-2Rβ ligand and/or an IL-2Rγc ligand can include moieties configured to maintain a circulating reservoir of a compound comprising an IL-2Rβ ligand and/or an IL-2Rγc ligand.

Compounds containing IL-2Rβ and/or IL-2Rγc ligands can include moieties configured to target IL-2R-directed immune stimulation of effector immune cells in tumors.

Compounds containing IL-2Rβ and/or IL-2Rγc ligands can include moieties designed to target specific immune cells, such as Treg cells.

Moieties can include compounds that are toxic to cells targeted by compounds containing IL-2Rβ ligands and/or IL-2Rγc ligands. Toxic moieties can be cleavable or otherwise activated, for example, by exposure to electromagnetic radiation. Toxic moieties can be activated, for example, by exposure to electromagnetic radiation such as visible radiation or ultraviolet radiation.
Compounds comprising IL-2Rβ ligands and/or IL-2Rγc ligands provided by the present disclosure can activate IL-2 receptors. Compounds comprising IL-2Rβ ligands and/or IL-2Rγc ligands provided by this disclosure can inhibit IL-2 receptors. Certain compounds comprising IL-2Rβ ligands and/or IL-2Rγc ligands provided by this disclosure bind IL-2Rβ subunits and/or IL-2Rγc subunits, and other compounds bind IL-2Rβ subunits. and/or can be prevented from binding to the IL-2Rγc subunit. Compounds comprising IL-2Rβ ligands and/or IL-2Rγc ligands reduce the efficacy of IL-2R agonists on cells that highly express IL-2Rβ subunits and/or IL-2Rγc subunits, or Can interfere with binding. Compounds containing IL-2Rβ ligands and/or IL-2Rγc ligands can desensitize cells to IL-2.

Compounds comprising IL-2Rβ ligands and/or IL-2Rγc ligands provided by this disclosure can include compounds that act as IL-2Rβγc agonists or IL-2Rαβγc agonists.

The IL-2Rβγc agonists or IL-2Rαβγc agonists provided by the present disclosure are for making single chain peptides comprising IL-2Rβ ligands, IL-2Rγc ligands, and in the case of IL-2Rαβγc agonists, IL-2Rα ligands. can include synthetic or recombinant peptides tandemly linked to The ligands can be in any order and separated by amino acid linkers. Synthetic peptides can include natural amino acids or peptides with natural amino acids, and suitable substitutions with non-natural amino acids. IL-2Rβγc agonists and IL-2Rαβγc agonists provided by this disclosure can be recombinant fusion proteins comprising IL-2Rγc ligand and IL-2Rβ ligand, and in the case of IL-2Rαβγc agonists, IL-2Rα ligand, and Fc Fusion partners can be proteins, IgG proteins, human serum albumin or other natural or engineered proteins, or hydrophilic biodegradable protein polymers. IL-2Rβγc agonists or IL-2Rαβγc agonists comprise one or more IL-2Rβ ligands and/or one or more IL-2Rγc ligands and, in the case of IL-2Rαβγc agonists, one or more IL-2Rα ligands can be done. IL-2Rβγc agonists or IL-2Rαβγc agonists can include IL-2Rβ ligands and IL-2Rγc ligands, and in the case of IL-2Rαβγc agonists can include IL-2Rα ligands, such as PEG or albumin binding moieties. It can further comprise one or more moieties selected to modify the pharmacokinetics of the IL-2R agonist.

IL-2Rβγc agonists can bind to IL-2Rβ subunits and IL-2Rγc subunits and can activate IL-2 receptors. The binding affinities (IC ₅₀ ) of IL-2Rβγc agonists for IL-2Rβ subunit and IL-2Rγc subunit are independently, e.g., less than 100 μM, less than 10 μM, less than 1 μM, less than 100 nM, less than 10 nM, or less than 1 nM can be IL-2Rβγc agonists can bind IL-2Rβ and IL-2Rγc competitively or non-competitively with IL-2.

IL-2Rαβγc agonists can bind IL-2Rα, IL-2Rβ, and IL-2Rγc and activate the IL-2 receptor. The binding affinity (IC ₅₀ ) of an IL-2Rαβγc agonist for IL-2Rα, IL-2Rβ, and IL-2Rγc can be, for example, less than 100 μM, less than 10 μM, less than 1 μM, less than 100 nM, less than 10 nM, or less than 1 nM. . IL-2Rαβγc agonists can bind IL-2Rα, IL-2Rβ and IL-2Rγc competitively or non-competitively with IL-2.

IL-2Rβγc agonists or IL-2Rαβγc agonists include IL-2Rβ ligands and IL-2Rγc ligands, and in the case of IL-2Rαβγc agonists include IL-2Rα ligands and express IL-2Rβ subunits and IL-2Rγc subunits. can be configured to activate more potently the cells that target cells, thereby enhancing the ability to differentially activate IL-2R expressed on the surface of different cell types by controlling the dose of the agonist. Facilitate. For example, primary human peripheral blood mononuclear cells (PBMC) expressing IL-2Rαβγc subunits phosphorylate transcription 5 (STAT5) when incubated with heteroisomeric compounds containing IL-2Rβ and IL-2Rγc ligands. . A heteromer can comprise an IL-2Rβ ligand, an IL-2Rγc ligand, and a linker, wherein the linker is configured such that the heteromer is an agonist of the IL-2 receptor. Linkers can include lengths that facilitate binding of IL-2Rβ and IL-2Rγc ligands to the IL-2 receptor. For example, the linker can have a length of 10 Å to 400 Å, 10 Å to 300 Å, 10 Å to 200 Å, 20 Å to 100 Å, 30 Å to 80 Å, or 40 Å to 60 Å. The linker can include chemical structures that facilitate simultaneous binding of IL-2Rβ and IL-2Rγc ligands to their respective IL-2 receptor subunits. For example, linkers can include peptides or carbohydrates.

IL-2Rβγc agonists or IL-2Rαβγc agonists can partially activate the IL-2 receptor. Partial activation refers to activation levels that are, for example, less than 75% of maximal activation, less than 50% of maximal activation, less than 25%, less than 10%, or less than 1% of maximal activation. Maximum activation (E _max ) is the amplitude of cellular signal (activation) achievable at high agonist concentrations, such as high concentrations of IL-2. Partial IL-2R agonists can be effective in modulating IL-2R response levels to activation of the IL-2Rβ and IL-2Rγc subunits among different IL-2R-expressing cell types. For example, different cell types are known to vary in the expression levels of each of the IL-2R subunits, IL-2Rα, IL-2Rβ, and IL-2Rγc, and exhibit different sensitivities to IL-2R agonists.

IL-2R agonists, including one or more IL-2Rα ligands, one or more IL-2Rβ ligands, and one or more IL-2Rγc ligands, are administered on cells expressing IL-2Rα subunits, such as Tregs. Increased binding and potency can be demonstrated.

IL-2Rαβγc agonists can include IL-2Rα ligands as well as modified IL-2Rβ and/or IL-2Rγc ligands. Modified IL-2Rβ and IL-2Rγc ligands can be selected or designed to bind and activate IL-2R, but with low or moderate affinity and potency for IL-2R. Such IL-2Rαβγc agonists may be useful between cells with high IL-2Rα expression and cells with low levels of IL-2Rα expression, e.g., Tregs with high IL-2Rα expression and low expression Teff cells with levels of IL-2Rα may have greater differential sensitivity for IL-2R activation.

IL-2Rβγc agonists or IL-2Rαβγc agonists comprise one or more IL-2Rβ ligands and/or one or more IL-2Rγc ligands and, in the case of IL-2Rαβγc agonists, one or more IL-2Rα ligands can be done. The presence of multiple IL-2Rα, IL-2Rβ, and/or IL-2Rγc ligands correlates with IL-2Rα, IL-2Rβ, and/or IL-2Rγc expression levels in cells with low levels of IL-2Rα, IL-2Rβ, and/or IL-2Rγc. Potency of IL-2R agonists can be preferentially increased on cells that highly express 2Rα, IL-2Rβ, and/or IL-2Rγc.

IL-2R agonists can include moieties that have additional pharmacological activities other than that mediated by activation of the IL-2 receptor. The pharmacological activity can be an activity that has synergistic therapeutic efficacy with the IL-2R agonist, or the pharmacological activity can be an activity that has non-synergistic therapeutic efficacy with the IL-2R agonist. For example, useful pharmacologically active moieties or molecules can include checkpoint inhibitors.

Compounds provided by this disclosure include IL-2Rβ antagonists and IL-2Rγc antagonists. IL-2R antagonists inhibit the binding of IL-2 and variants and modified forms thereof to the IL-2Rβ subunit or to the IL-2Rγc subunit, and/or inhibit IL-2 activity of the IL-2 receptor. Compounds comprising IL-2Rβ ligands or IL-2Rγc ligands that reduce cytotoxicity.

IL-2Rβ and IL-2Rγc antagonists can attenuate the sensitivity of cells expressing IL-2Rβ and/or IL-2Rγc subunits to activation by IL-2 or variants and modified forms thereof. .

IL-2Rβ antagonists and IL-2Rγc antagonists can include compounds that have more than one IL-2Rβ ligand or more than one IL-2Rγc ligand, and can inhibit IL-2 receptors competitively or non-competitively with IL-2. can bind to the body.

IL-2Rβ antagonists and IL-2Rγc antagonists can comprise one or more IL-2Rβ ligands or one or more IL-2Rγc ligands and moieties with useful pharmacological activity. A moiety can exhibit pharmacological activity that is synergistic with IL-2R inhibition or not synergistic with IL-2R inhibition.

IL-2Rβ antagonists and IL-2Rγc antagonists can further comprise recombinant fusion proteins.

Compounds provided by this disclosure include IL-2R antagonists.

IL-2R antagonists can include IL-2Rα and IL-2Rβ ligands, IL-2Rα and IL-2Rγc ligands, or IL-2Rβ and IL-2Rγc ligands.

IL-2R antagonists include compounds that bind to either the IL-2Rβ or IL-2Rγc subunits and inhibit activation of the IL-2 receptor.

IL-2R antagonists include compounds that bind to the IL-2Rβ and IL-2Rγc subunits and inhibit activation of the IL-2 receptor, IL-2Rβ and IL-2Rγc ligands interacting with IL-2 receptors. Constructed so as not to activate the body. Such compounds are high affinity antagonists of IL-2R activation and the presence of both IL-2Rβ and IL-2Rγc ligands enhances the potency of IL-2R antagonists.

IL-2R antagonists include compounds containing IL-2Rβ ligands and IL-2Rγc ligands, which are configured to exhibit partial activation of the IL-2 receptor. These compounds are examples of partial IL-2R antagonists. Such compounds are useful for modulating the level of cellular response to IL-2R agonists between cells with different expression levels of IL-2R subunits. The use of partial IL-2R agonists/antagonists can modulate the response of cells to IL-2R agonists among cells with different expression levels of IL-2Rβ and/or IL-2Rγc subunits.

IL-2R antagonists can include one or more IL-2Rβ and/or IL-2Rγc ligands. IL-2R antagonists can be synthetic or recombinant, can be peptides or polypeptides IL-2R ligands can be attached in any order, any orientation, and can be attached to a linker. Linkers can include natural and/or non-natural amino acids and/or non-peptidyl structures.

IL-2R antagonists can be chemically modified to include moieties that affect the pharmacokinetics of IL-2R antagonists, such as, for example, PEG and albumin binding moieties.

IL-2R antagonists can further include recombinant fusion proteins.

Compounds containing IL-2Rβ ligands and/or IL-2Rγc ligands include diagnostic reagents. As diagnostic agents, compounds containing IL-2Rβ ligands and/or IL-2Rγc ligands can be used to detect and/or measure cells expressing IL-2Rβ subunits and/or IL-2Rγc subunits. . Compounds can be used to determine the level of IL-2Rβ and/or IL-2Rγc expression in a cell, cell population, or tissue. Compounds can be used to assess the binding affinity of IL-2Rβ subunits and/or IL-2Rγc subunits to cells or cell populations. A compound may be used to determine a particular cell type based, for example, on IL-2Rβ and/or IL-2Rγc expression levels.

Compounds may be useful for in vitro and in vivo diagnostics.

Diagnostic compounds containing IL-2Rβ and/or IL-2Rγc ligands can include detectable markers. Detectable markers may be cleavable or non-cleavable.

Detectable markers can include, for example, radioactive labels, fluorescent labels, enzymatic labels.

Cells expressing IL-2Rβ subunits and/or IL-2Rγc subunits in biological samples, such as patient blood samples, and/or using diagnostic compounds containing IL-2Rβ ligands and/or IL-2Rγc ligands. Alternatively, expression levels of cells expressing IL-2Rβ subunit and/or IL-2Rγc subunit can be measured. Measurements can be made, for example, using flow cytometry. The number of cells expressing the IL-2Rβ subunit and/or the IL-2Rγc subunit and/or the expression level of the IL-2Rβ subunit and/or the IL-2Rγc subunit is an indication of the patient's disease or the drug for the patient's disease. When correlated with physiologically significant parameters, it can be used to inform disease treatment. For example, IL-2Rβ subunit and/or IL-2Rγc subunit expression levels are above or below therapeutically significant thresholds for a particular disease, IL-2Rβ ligands and/or Alternatively, a compound comprising an IL-2Rγc ligand can be administered to the patient to treat the disease.

Compounds containing IL-2Rβ ligands and/or IL-2Rγc ligands can be bound to solid supports. Based on the ability of a compound to bind to the IL-2Rβ subunit and/or IL-2Rγc subunit, the compound may be used as a reagent for detecting IL-2Rβ subunit and/or IL-2Rγc subunit, for example, It can be used as a natural reagent in cells, fixed cells, biofluids, tissue homogenates, purifications, and biomaterials. Additionally, based on their ability to bind the IL-2Rβ subunit and/or the IL-2Rγc subunit, the peptides of the invention can be tested, for example, by in situ staining, FACS (fluorescence activated cell sorting), Western blotting, and ELISA. Additionally, the compounds provided by the present disclosure can be used for receptor purification or to purify cells that express IL-2Rβ subunit and/or IL-2Rγc subunit on the cell surface. obtain.

Compounds comprising IL-2Rβ ligands and/or IL-2Rγc ligands provided by this disclosure can also be used as reagents for various medical research and diagnostic applications. Such uses include, for example, use as calibration standards to quantify the activity of candidate IL-2R agonists or IL-2R antagonists in functional assays, to maintain proliferation and growth of IL-2 dependent cell lines. (3) use in structural elucidation of IL-2 receptor by co-crystallization, use to investigate mechanisms of IL-2 signaling/receptor activation, and where IL-2 receptor is involved Other research and diagnostic uses are included.

Assessing a single patient's response to treatment and qualifying a patient for optimal treatment is one of the greatest challenges in modern medicine and is relevant to the trend of personalized medicine. Compounds comprising IL-2Rβ ligands and/or IL-2Rγc ligands can have target selectivity for diseases in which cells associated with disease etiology express IL-2Rβ subunits and/or IL-2Rγc subunits. . For example, using a compound containing IL-2Rβ and/or IL-2Rγc ligand labeled for positron emission tomography (PET) or single photon emission computed tomography (SPECT), a single case-by-case Targets of treatment can be predicted based on patient analysis of cases, so that subjects who are not expected to benefit from treatment with therapeutic compounds that affect the activity of the IL-2Rβ subunit and/or IL-2Rγc subunit. can be excluded. PET/SPECT scans using compounds containing IL-2Rβ and/or IL-2Rγc ligands provide three-dimensional distribution maps when correlated with concentrations of compounds containing IL-2Rβ and/or IL-2Rγc ligands. can then be used for macroscopic dose calculation.

Compounds containing IL-2Rβ ligands and/or IL-2Rγc ligands can include one or more imaging agents. IL-2Rβ ligands and/or IL-2Rγc ligands can direct and localize the compounds to cells, cell populations and tissues that express IL-2Rβ subunits and/or IL-2Rγc subunits. Imaging compounds can include one or more imaging agents such as radiolabels, fluorescent labels, enzymatic labels, or PET imaging agents.

Using an imaging agent, the number of cells expressing IL-2Rβ subunit and/or IL-2Rγc subunit, the expression level of cells expressing IL-2Rβ subunit and/or IL-2Rγc subunit, or IL a specific IL-2Rβ ligand and/or IL-2Rγc and/or IL-2Rβ of a -2Rβ subunit and/or IL-2Rγc subunit, such as IL-2Rβ subunit and/or IL-2Rγc subunit characteristics Affinities for compounds containing ligands and/or IL-2Rγc ligands can be determined. Imaging agents can be used, for example, to assess cancer cells that express IL-2Rβ subunits and/or IL-2Rγc subunits, or to assess Treg and/or Teff cells.

The label can be detected to determine the biodistribution of the compound in a patient or to assess potential therapeutic efficacy. For example, tumors that express high levels of the IL-2R receptor and/or IL-2Rβ subunit and/or IL-2Rγc subunit are treated with IL-2Rβ ligands and/or IL-2Rγc ligands provided by the present disclosure. may be attractive targets for therapeutic compounds including.

Imaging agents can be used to assess cells expressing IL-2Rβ subunits and/or IL-2Rγc subunits before, during, and/or after treatment.

Imaging agents comprising IL-2Rβ and/or IL-2Rγc ligands may further comprise moieties capable of binding to cell surfaces, particularly proteins expressed on cell surfaces. Proteins can be indicative of specific cell types and are referred to as cell surface markers. cells expressing both an IL-2Rβ subunit and/or an IL-2Rγc subunit and a cell surface marker using an imaging agent comprising both an IL-2Rβ ligand and/or an IL-2Rγc ligand and a cell surface marker; Cell populations and/or tissues can be evaluated. The assessment includes the number of cells expressing both the IL-2Rβ subunit and/or IL-2Rγc subunit and the cell surface marker, the expression level of the IL-2Rβ subunit and/or the IL-2Rγc subunit and the cell surface marker, and/or determining the affinity of the imaging agent for the IL-2Rβ subunit and/or the IL-2Rγc subunit and/or the cell surface marker.

Imaging agents can be used to assess cells expressing IL-2Rβ subunits and/or IL-2Rγc subunits and cell surface markers before, during, and/or after treatment.

As an illustration, T-cell infiltration of tumor lesions is a known prognostic factor in several tumor types and is used as a therapeutic mechanism in some of these tumor types. For example, in metastatic melanoma, treatment with immune checkpoint inhibitors induces clinical benefit in approximately 30-50% of patients. Tumor-infiltrating T cells express the IL-2 receptor on their surface. These T cells can therefore be visualized by molecular imaging using compounds containing IL-2Rβ and/or IL-2Rγc ligands and radiolabels such as PET tracers.

As another example, IL-2 is synthesized and secreted by activated T lymphocytes, particularly CD8+ CTL and CD4+ Th1 lymphocytes. T lymphocyte activation is observed in many types of inflammatory diseases such as inflammatory degenerative diseases, graft rejection, tumor inflammation, organ-specific autoimmune diseases, and lipoinflammatory insulin resistance. IL-2 binds with high affinity to plasma membrane IL-2 receptors that are predominantly expressed on the cell surface of activated T lymphocytes. Thus, PET imaging of activated T lymphocytes with radiolabeled IL-2Rβ and/or IL-2Rγc ligands provides a dynamic approach in vivo in studying immune cell infiltration in these inflammatory diseases.

Compounds provided by the present disclosure can be labeled. Labeled compounds may be useful in diagnosis.

Compounds comprising IL-2Rβ ligands and/or IL-2Rγc ligands and selective IL-2Rβγc agonists provided by this disclosure can be labeled with a detectable marker. A label can be used to determine the biodistribution of a compound in a patient or to assess potential therapeutic efficacy. For example, tumors that express high levels of the IL-2R receptor are attractive targets for selective IL-2Rβγc agonists and compounds containing IL-2Rβ and/or IL-2Rγc ligands provided by the present disclosure. can be

Accordingly, compounds provided by the present disclosure include labeled compounds. The labeling compound may be a detectable marker, such as a radiolabeled amino acid, or the binding of a biotinyl moiety to a polypeptide, the bound biotinyl moiety being labeled with avidin (e.g., a fluorescent marker or optical methods or streptavidin), which contains enzymatic activity that can be detected by colorimetric methods. Various methods of labeling polypeptides and glycoproteins are known in the art and can be used. Examples of labels for polypeptides include, for example, radioisotopes such as ³ H, ¹⁴ C, ³⁵ S, ¹²⁵ I, and ¹³¹ I; fluorescent labels such as FITC, rhodamine, and lanthanideline; horseradish peroxidase; Enzymatic labels such as galactosidase, luciferase, and alkaline phosphatase, biotinyl groups, predetermined polypeptide epitopes recognized by secondary reporters such as leucine zipper pair sequences, binding sites for secondary antibodies, metal ligands, and epitope tags. . Labels can be attached by spacer arms of various lengths to reduce potential steric hindrance.

Compounds containing IL-2Rβ ligands and/or IL-2Rγc ligands can include cell-specific targeting moieties or molecules.

Cell-specific targeting moieties can include moieties that have affinity for components on the surface of cells such as receptors, proteins, or epitopes. A moiety can include, for example, a ligand or an antibody that has an affinity for a cell surface component.

A targeting moiety can induce and concentrate compounds, including IL-2Rβ and/or IL-2Rγc ligands, in cells, cell populations, or tissues targeted by the targeting moiety.

A targeting moiety can enhance the potency of IL-2R agonism or IL-2R antagonism on a targeted cell or cell population.

A targeting moiety can provide a differential response to IL-2R agonism or IL-2R antagonism between targeted cells and cells not targeted by the targeting moiety.

The targeting moiety provides a differential response to IL-2R agonism or IL-2R antagonism between cells with high expression levels of the targeting component and cells with lower expression levels of the targeting component. can do.

Compounds comprising IL-2Rβ ligands and/or IL-2Rγc ligands may further comprise bioactive moieties or molecules. A compound comprising an IL-2Rβ ligand and/or an IL-2Rγc ligand is used to transfer a bioactive portion or molecule to a cell, cell population, or tissue expressing an IL-2Rβ subunit and/or an IL-2Rγc subunit. can be delivered to

A bioactive portion or molecule can be non-cleavable and can exert bioactivity when bound to a compound that includes an IL-2Rβ ligand and/or an IL-2Rγc ligand.

A bioactive portion or molecule may be cleavable. A moiety may be cleavable by any suitable mechanism, such as pH, enzymatic, thermal, and/or electromagnetic mechanisms. Electromagnetic mechanisms include, for example, exposing the compound to infrared, visible, or ultraviolet radiation, wherein the bioactive moiety binds IL-2Rβ ligand and/or IL-2Rβ ligand and/or IL-2Rβ ligand through a labile moiety that can be cleaved by radiation. It binds to compounds containing -2Rγc ligands.

A bioactive molecule can be non-cleavable, but otherwise activatable, eg, activatable by exposure to electromagnetic radiation.

IL-2Rβ and/or IL-2Rγc ligands may be selected to have enhanced binding to IL-2Rβ and/or IL-2Rγc subunits at a particular pH. For example, pH-selective IL-2Rβ ligands and/or IL-2Rγc ligands are more sensitive to IL-2Rβ subunits and/or IL-2Rγc subunits, respectively, at low pH comparable to that of the solid tumor microenvironment. It can have high affinity. Using compounds comprising low pH selective IL-2Rβ ligands and/or IL-2Rγc ligands, IL-2Rβ subunit and/or IL- Cells in low pH environments that express the 2Rγc subunit can be preferentially activated.

Accordingly, compounds comprising selective IL-2Rβ ligands and/or IL-2Rγc ligands, such as pH-selective IL-2Rβ ligands and/or pH-selective IL-2Rγc ligands, can be used along with other pH-selective biologically active moieties and molecules. can be used.

The bioactive portion or bioactive molecule itself can be selective for a particular cell population. For example, a bioactive portion or molecule can exhibit greater or lesser affinity, potency, and/or activity in cells targeted by selective IL-2Rβ and/or IL-2Rγc ligands. can. For example, bioactive portions or molecules can exhibit greater bioactivity in low pH tumor microenvironments when targeted by pH-selective IL-2Rβ and/or IL-2Rγc ligands. In this example, the bioactive moiety activates cells located in a low pH tumor microenvironment that express IL-2Rβ and/or IL-2Rγc subunits via pH-selective IL-2Rβ and/or IL-2Rγc ligands. set to target. Thus, the activity of pH-selective bioactive moieties is enhanced in the low-pH tumor microenvironment.

Compounds comprising IL-2Rβ ligands and/or IL-2Rγc ligands may further comprise cytotoxic moieties or molecules. Such compounds can be used to deliver cytotoxic moieties or compounds to cells that express IL-2Rβ subunits and/or IL-2Rγc subunits, such as T cells. A cytotoxic moiety or molecule can exert cytotoxicity when bound to a compound or can be cleavable, and a moiety or molecule can be cytotoxic when released from a compound. Alternatively, the cytotoxic moiety can be activated by electromagnetic radiation.

Cytotoxic moieties or molecules can be used to deplete cells expressing the targeted IL-2Rβ and/or IL-2Rγc subunits.

An IL-2Rβ ligand and/or IL-2Rγc ligand-containing cytotoxic compound may have more than one IL-2Rβ ligand and/or more than one IL-2Rγc ligand, whereby IL-2Rβ subunits and/or higher affinity for cells, cell populations and tissues that highly express the IL-2Rβ subunit and/or the IL-2Rγc subunit compared to cells that express low levels of the IL-2Rγc subunit can indicate a specificity and/or selectivity.

IL-2Rβ ligand and/or IL-2Rγc ligand-containing cytotoxic compounds can further comprise a cell surface targeting moiety. Such cytotoxic compounds can exhibit enhanced efficacy against cells, cell populations and tissues expressing IL-2Rβ subunits and/or IL-2Rγc subunits and surface targeting moieties. .

Examples of suitable cytotoxic molecules include antimicrotubule agents, alkylating agents, and DNA minor groove binding agents.

Compounds that include IL-2Rβ and/or IL-2Rγc ligands may further include moieties that have useful pharmacological activities unrelated to IL-2 activity.

The pharmacological moiety can function synergistically with IL-2R agonist activity, or it can function synergistically with IL-2R antagonist activity, or it can function synergistically with IL-2Rβ subunit and/or Synergy with IL-2Rγc subunit activity may not be shown.

Examples of suitable pharmacological moieties include antibodies and antibody fragments that are inhibitors of checkpoint molecules, pro- and anti-apoptotic molecules, cytotoxic molecules, chemokine agonists, chemokine antagonists, cytokines, growth factors. and other cell surface receptors, as well as ligands and inhibitors of cell surface adhesion molecules such as integrins.

Peptides provided by the present disclosure can be synthesized by methods known in the art, for example, by using standard solid phase techniques.

Peptides containing IL-2Rβ and/or IL-2Rγc ligands provided by this disclosure can be modified, for example, by phosphorylation and by other methods known in the art. Thus, the peptides provided by this disclosure can also serve as a basis for preparing peptidomimetics with similar biological activity.

A variety of techniques for constructing peptidomimetics that possess the same or similar desired biological activities as the corresponding peptides, but with advantageous activities over the peptides with respect to solubility, stability, and susceptibility to hydrolysis and proteolysis. is available.

Selective IL-2Rβγc agonists and compounds comprising IL-2Rβ ligands and/or IL-2Rγc ligands provided by the present disclosure can be administered intradermally, intramuscularly, intraperitoneally, intravenously, subcutaneously, intranasally, epidurally, It can be incorporated into pharmaceutical compositions that are administered to a patient by any suitable route of administration, including oral, oral, sublingual, intracerebral, intravaginal, transdermal, rectal, inhalation, or topical. In certain embodiments, pharmaceutical compositions provided by this disclosure are injectable formulations. A pharmaceutical composition provided by the present disclosure can be an injectable intravenous formulation. Pharmaceutical compositions provided by the present disclosure can be oral formulations. An oral formulation can be an oral dosage form. Pharmaceutical compositions may be formulated for intravenous or subcutaneous administration.

Pharmaceutical compositions provided by the present disclosure include a therapeutically effective amount of a selective IL-2Rβγc agonist, IL-2Rβ ligand and/or IL-2Rγc ligand to provide a composition for proper administration to a patient. or a pharmaceutically acceptable salt of any of the foregoing, and a suitable amount of one or more pharmaceutically acceptable vehicles. Suitable pharmaceutical vehicles and methods of preparing pharmaceutical compositions are described in the art.

Compounds containing selective IL-2Rβγc agonists or IL-2Rβ ligands and/or IL-2Rγc ligands can be administered by intravenous injection. Forms suitable for injection include sterile aqueous solutions or dispersions of selective IL-2Rβγc agonists or compounds, including IL-2Rβ ligands and/or IL-2Rγc ligands. Compounds containing selective IL-2Rβγc agonists or IL-2Rβ ligands and/or IL-2Rγc ligands can be formulated in physiological buffers. Prior to administration, a compound comprising a selective IL-2Rβγc agonist, an IL-2Rβ ligand and/or an IL-2Rγc ligand, or a pharmaceutically acceptable salt of any of the foregoing, is treated with an antibacterial or antifungal agent, For example, it may be sterilized by any art-recognized technique, including the addition of parabens, chlorobutanol, phenol, sorbic acid, thymersol, and the like. A selective IL-2Rβγc agonist, an IL-2Rβ ligand and/or a compound comprising an IL-2Rγc ligand, or a pharmaceutically acceptable salt of any of the foregoing, is sterilized by filtration prior to administration to a subject; The need for additional sterilants may thereby be minimized or eliminated. Injection volumes of compounds comprising selective IL-2Rβγc agonists or IL-2Rβ ligands and/or IL-2Rγc ligands are about 0.01 mL to about 10 mL, about 0.1 mL to about 10 mL, about 0.1 mL to about 5 mL, and in certain embodiments, from about 1 mL to about 5 mL.

Pharmaceutical compositions comprise a therapeutically effective amount of a compound comprising one or more selective IL-2Rβγc agonists or IL-2Rβ ligands and/or IL-2Rγc ligands in purified form together with a suitable amount of a pharmaceutically acceptable vehicle. to provide the patient with a form for proper administration. When administered to a patient, the compounds and pharmaceutically acceptable vehicles are preferably sterile. Water is a preferred vehicle when the compound is administered intravenously. Saline solutions and aqueous dextrose and glycerol solutions can also be employed as liquid vehicles, particularly for injectable solutions. Suitable pharmaceutical vehicles 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. Also included are excipients such as , water, ethanol, and the like. The pharmaceutical composition can also contain minor amounts of wetting or emulsifying agents, or pH buffering agents. Additionally, adjuvants, stabilizers, thickeners, lubricants, and colorants may be used.

A pharmaceutical composition containing a compound can be manufactured by conventional mixing, dissolving, granulating, floating, emulsifying, encapsulating, entrapping, or lyophilizing processes. Pharmaceutical compositions can be formulated in a conventional manner using one or more physiologically acceptable carriers, diluents, excipients or adjuvants, which are pharmaceutically acceptable preparations. facilitates processing of the compound into Proper formulation is dependent on the route of administration chosen. The pharmaceutical compositions provided by this disclosure can take the form of solutions, suspensions, emulsions, or any other form suitable for use. Examples of suitable pharmaceutical vehicles are described in the art.

For parenteral administration, compounds containing selective IL-2Rβγc agonists and IL-2Rβ ligands and/or IL-2Rγc ligands may be incorporated into a solution or suspension. Parenteral administration refers to administration by injection, eg, intravenous, intravesical, intrathecal, intrapleural, intratumoral, subcutaneous, or intraperitoneal injection, or intravesical administration. Selective IL-2Rβγc agonists or compounds, including IL-2Rβ ligands and/or IL-2Rγc ligands, can be administered intravenously.

The solutions or suspensions may also contain at least one of the following adjuvants: sterile dilutions such as water for injection, saline, fixed oils, polyethylene glycols, glycerol, propylene glycol or other synthetic solvents. antioxidants such as ascorbic acid or sodium bisulfite; buffers such as acetate, citrate or phosphate; and agents for adjusting osmotic pressure such as sodium chloride or dextrose. A parenteral preparation can be enclosed in ampoules, disposable syringes or multiple dose containers made of glass or plastic.

Assessing a single patient's response to treatment and qualifying a patient for the best treatment is one of the greatest challenges in modern medicine and is relevant to the trend of personalized medicine. Compounds comprising selective IL-2Rβγc agonists or IL-2Rβ ligands and/or IL-2Rγc ligands can have target selectivity for particular cancers. using selective IL-2Rβγc agonists and compounds, including radiolabeled IL-2Rβ and/or IL-2Rγc ligands for Positron Emission Tomography (PET) or Single Photon Emission Tomography (SPECT), Targets of treatment can be predicted based on single case-by-case patient analysis, thus excluding subjects who are not expected to benefit from treatment. PET/SPECT scans using compounds containing radiolabeled selective IL-2Rβγc agonists or IL-2Rβ ligands and/or IL-2Rγc ligands showed concentration-selective IL-2Rβγc agonists or IL-2Rβ ligands and/or IL-2Rγc When correlated with ligand-containing compounds, a three-dimensional distribution map can be provided, which can then be used for macroscopic dose calculations.

Thus, assaying and using selective IL-2Rβγc agonists, IL-2Rβ ligands and/or compounds comprising IL-2Rγc ligands, and/or pharmaceutical compositions thereof for therapy is within the capabilities of those skilled in the art. is within.

Selective IL-2Rβγc agonists, IL-2Rβ ligands and/or compounds comprising IL-2Rγc ligands, and/or pharmaceutical compositions thereof will generally be used in amounts effective to achieve their intended purpose. can. For use in treating diseases such as cancer, compounds comprising selective IL-2Rβγc agonists, IL-2Rβ ligands and/or IL-2Rγc ligands, and/or pharmaceutical compositions thereof are administered in therapeutically effective amounts can be administered or applied.

compounds comprising selective IL-2Rβγc agonists, IL-2Rβ ligands and/or IL-2Rγc ligands, and/or any of the foregoing, which would be effective in treating the particular disorders or conditions disclosed herein The amount of such pharmaceutical composition will depend in part on the nature of the disorder or condition, and can be determined by standard clinical techniques known in the art. In addition, in vitro or in vivo assays may optionally be employed to help identify optimal dosage ranges. The amount of a selective IL-2Rβγc agonist, an IL-2Rβ ligand and/or a compound comprising an IL-2Rγc ligand, and/or any of the foregoing pharmaceutical compositions may vary depending on, among other things, the subject to be treated, the weight of the subject, the degree of affliction. It depends on the severity, method of administration, and judgment of the prescribing physician.

Compounds comprising selective IL-2Rβγc agonists or IL-2Rβ ligands and/or IL-2Rγc ligands can be assayed in vitro and in vivo for the desired therapeutic activity prior to use in humans. For example, in vitro assays can be used to determine whether administration of a particular compound or combination of compounds is preferred. Compounds can also be demonstrated to be effective and safe using animal model systems.

In certain embodiments, a therapeutically effective amount of a selective IL-2Rβγc agonist, IL-2Rβ ligand and/or compound comprising an IL-2Rγc ligand, and/or the pharmaceutical composition of any of the foregoing substantially comprises provide therapeutic benefit without causing significant toxicity. Toxicity of selective IL-2Rβγc agonists, IL-2Rβ ligands and/or compounds comprising IL-2Rγc ligands, and/or pharmaceutical compositions of any of the foregoing is determined using standard pharmaceutical procedures. and can be readily ascertained by those skilled in the art. The dose ratio between toxic and therapeutic effects is the therapeutic index. In certain embodiments, selective IL-2Rβγc agonists, IL-2Rβ ligands and/or compounds comprising IL-2Rγc ligands, and/or pharmaceutical compositions of any of the foregoing are used in the treatment of diseases and disorders. It exhibits a particularly high therapeutic index. The dose of a selective IL-2Rβγc agonist, IL-2Rβ ligand and/or compound comprising an IL-2Rγc ligand, and/or pharmaceutical composition of any of the foregoing is circulating concentrations that include an effective dose with minimal toxicity. Within range.

Compounds or pharmaceutical compositions thereof comprising IL-2Rβ and/or IL-2Rγc ligands provided by this disclosure can be included in kits that can be used to administer the compounds to a patient for therapeutic purposes. A kit comprises a pharmaceutical composition comprising a selective IL-2Rβγc agonist, or a compound comprising an IL-2Rβ ligand and/or an IL-2Rγc ligand provided by the present disclosure suitable for administration to a patient, and a pharmaceutical composition to a patient. may include instructions for administering to. The kit can be a kit for treating cancer. Kits for use in treating cancer in a patient include selective IL-2Rβγc agonists or compounds comprising IL-2Rβ ligands and/or IL-2Rγc ligands provided by the present disclosure, for administering compounds and instructions for administering the compound to the patient.

Pharmaceutical compositions can be included in a container, pack, or dispenser together with instructions for administration.

Instructions supplied with the kit may be printed and/or supplied, for example, as an electronic readable medium, videocassette, audiotape, flash memory device, or may be published on an Internet website, or may be electronic. It may be distributed to the patient and/or healthcare provider as a communication.

Selective IL-2Rβγc agonists and compounds, including IL-2Rβ ligands and/or IL-2Rγc ligands provided by this disclosure, can be used to treat cancer in patients. A cancer can be, for example, a solid tumor or a metastasis.

Selective IL-2Rβγc agonists and compounds, including IL-2Rβ ligands and/or IL-2Rγc ligands, or pharmaceutical compositions thereof, provided by the present disclosure are treated by activating or inhibiting the IL-2 receptor. can be administered to treat cancers known to Selective IL-2Rβγc agonists and compounds, including IL-2Rβ ligands and/or IL-2Rγc ligands, or pharmaceutical compositions thereof, provided by the present disclosure are treated by activation or inhibition of the IL-2Rβγc subunit. are known and may be administered to treat cancers where co-activation of IL-2Rα subunits compromises therapeutic efficacy and/or minimizes undesirable side effects.

Selective IL-2Rβγc agonists and compounds, including IL-2Rβ ligands and/or IL-2Rγc ligands or pharmaceutical compositions thereof, provided by the present disclosure may be used, for example, to treat one or more of the following cancers: Can be used for: acute lymphoblastic leukemia, acute myelogenous leukemia, adrenocortical carcinoma, appendiceal carcinoma, astrocytoma, atypical terato/striated tumor, basal cell carcinoma (non-melanoma), B-cell lymphoma, bladder cancer, bone cancer, brain and spinal cord tumor, brain stem cancer, brain tumor, breast cancer, bronchial tumor, Burkitt's lymphoma, cancerous tumor, head and neck cancer, central nervous system embryonic tumor, cerebellar astrocytoma , cerebral astrocytoma/malignant glioma, uterine cancer, nodule, chronic lymphocytic lymphocytic leukemia, chronic myelogenous leukemia, colorectal cancer, craniopharyngioma, cutaneous T-cell lymphoma, deplastic small round cell tumor, Ductal carcinoma, dye carcinoma, endocrine pancreatic tumor (pancreatic islet cell tumor), endometrial cancer, renal pyeloma, esophageal cancer, sensory neuroblastoma, Ewing family tumor, extracranial germ cell tumor, extrahepatic cholangiocarcinoma, gallbladder Cancer, gastric cancer, gastrointestinal cancer, gastric cancer, carcinoid tumor, gastrointestinal stromal tumor, gestational neutrophilic tumor, glioblastoma, glioma, hairy cell leukemia, head and neck cancer, cardiac cancer, lymphoid hematopoiesis tumor, hepatocellular carcinoma, Hodgkin's lymphoma, subpharyngeal carcinoma, hypothalamic and visual pathway glioma, ID-related lymphoma, intraocular melanoma, pancreatic cell tumor, Kaposi's sarcoma, renal cancer, Langerhans cell-stimulating cells, laryngeal cancer, leukemia , lip and oral cancer, male breast cancer, malignant fibrous histiocytoma, malignant germ cell tumor, malignant mesothelioma, medulloblastoma, melanoma, Merkel cell carcinoma, mesothelioma, oral cancer, multiple endocrine neoplasms syndrome, multiple myeloma, mycosis fungoides, myelodysplastic syndrome, myeloproliferative neoplasia, nasal and sinus sinus cancer, nasopharyngeal carcinoma, neuroblastoma, non-Hodgkin's lymphoma, non-small cell lung cancer, oral cancer , oropharyngeal cancer, osteosarcoma, ovarian cancer, ovarian epithelial cancer, ovarian germ cell tumor, ovarian low malignant potential tumor, pancreatic cancer, pancreatic neuroendocrine tumor (islet cell tumor), papilloma, accessory glioma, paranasal sinus and nasal cavity carcinoma, parathyroid carcinoma, penile carcinoma, pharyngeal carcinoma, peripheral melanocytic carcinoma, cerebellar germ cell tumor, cerebellar tumor pinoblastoma and supratendinous primitive neuroectodermal tumor, pituitary tumor, plasma cell neoplasm/multifocal Myeloma, pleuropulmonary blastoma, pregnancy and breast cancer, primary central nervous system lymphoma, primary liver cancer, primary metastatic squamous neck cancer (including occult), prostate cancer, rectal cancer, renal cell carcinoma , renal pelvic and ureteral cancer, respiratory cancer, retinoblastoma, rhabdomyosarcoma, salivary gland carcinoma, sarcoma, Sézary syndrome, skin cancer, skin cancer, small bowel cancer, soft tissue sarcoma, squamous cell carcinoma (non-melanoma) ), gastric cancer, supratendinous primitive neuroectodermal tumor, T-cell lymphoma, testicular cancer, throat cancer, thyroid and thyroid cancer, transitional cell carcinoma, urethral cancer, uterine sarcoma, vaginal cancer, visual pathway and hypothalamic glioma, Vulvar cancer, Waldenstrom's macroglobulinemia, Wilms tumor, and systemic and central metastases of any of the foregoing.

Selective IL-2Rβγc agonists and compounds comprising IL-2Rβ and/or IL-2Rγc ligands provided by the present disclosure, or pharmaceutical compositions of any of the foregoing, can be used to treat solid tumors. can.

Selective IL-2Rβγc agonists, compounds comprising IL-2Rβ ligands and/or IL-2Rγc ligands provided by the present disclosure, or pharmaceutical compositions of any of the foregoing may be used to treat, for example, the following cancers: The cancers include primary adult and infant brain and CNS cancers including glioblastoma (GBM) and astrocytoma, skin cancer including melanoma, small cell lung cancer lung cancer, including non-small cell lung cancer (NSCLC), and large cell lung cancer, breast cancer, including triple-negative breast cancer (TNBC), hematologic cancers, including myelodysplastic syndrome (MDS), multiple myeloma (MM), and acute myeloma leukemia (AML), prostate cancer including castration-resistant prostate cancer (CRPC), hepatocellular carcinoma (HCC), esophageal and gastric cancer, and any systemic and central metastasis of any of the foregoing.

The amount of a selective IL-2Rβγc agonist, IL-2Rβ ligand and/or compound comprising an IL-2Rγc ligand provided by the present disclosure, or any of the foregoing pharmaceutical compositions that would be effective in treating cancer is It will depend, at least in part, on the nature of the disease and can be determined by standard clinical techniques known in the art. In addition, in vitro or in vivo assays may be employed to help identify optimal dosage ranges. Dosage regimens and intervals can also be determined by methods known to those of ordinary skill in the art. The amount of IL-2Rβ ligand and/or selective IL-2Rβγc agonist or compound comprising an IL-2Rγc ligand provided by the present disclosure to be administered may vary depending on, among other things, the subject to be treated, the weight of the subject, the severity of the disease, It may depend on the route of administration and the judgment of the prescribing physician.

For systemic administration, a therapeutically effective dose can be estimated initially from in vitro assays. Initial doses can also be estimated from in vivo data, such as animal models, using techniques known in the art. Such information can be used to more accurately determine useful doses in humans. One skilled in the art may optimize administration to humans based on animal data.

The dose and appropriate dosing interval of an IL-2Rβ ligand and/or a selective IL-2Rβγc agonist or compound comprising an IL-2Rγc ligand provided by the present disclosure are selected such that, in certain embodiments, a minimal adverse concentration is exceeded. A sustained therapeutically effective concentration of selective IL-2Rβγc agonists or compounds, including IL-2Rβ ligands and/or IL-2Rγc ligands provided by the present disclosure, can be maintained in the patient's blood without any effort.

In certain embodiments, pharmaceutical compositions comprising selective IL-2Rβγc agonists or compounds comprising IL-2Rβ ligands and/or IL-2Rγc ligands provided by the present disclosure are administered, for example, twice a week. It can be administered once a week, once every three weeks, once every four weeks, once every five weeks, or once every six weeks. Dosing may be provided alone or in combination with other drugs and may continue as long as necessary for effective treatment of the disease. Dosing can also be carried out using continuous or semi-continuous administration over a period of time. Dosing includes administering a pharmaceutical composition to a mammal, eg, a human, in a fed or fasted state.

Pharmaceutical compositions can be administered in single or multiple dosage forms, or as continuous or cumulative doses over a period of time. When multiple dosage forms are used, the amount of a selective IL-2Rβγc agonist or compound comprising an IL-2Rβ ligand and/or IL-2Rγc ligand provided by the present disclosure contained in each of the multiple dosage forms is , may be the same or different.

A suitable daily dosage range for administration is in the range of about 2 μg to about 200 mg of IL-2Rβ ligand and/or selective IL-2Rβγc agonists or compounds, including IL-2Rγc ligands provided by the present disclosure, per kg of body weight. could be.

A suitable daily dosage range for administration is about 1 μg of selective IL-2Rβγc agonists or compounds, including IL-2Rβ ligands and/or IL-2Rγc ligands provided by the present disclosure, per square meter (m ² ) of body surface. may range from to about 50 mg.

Selective IL-2Rβγc agonists or compounds, including IL-2Rβ ligands and/or IL-2Rγc ligands provided by the present disclosure, can be used to treat cancer in a subject, for example, in amounts of 0.001 mg/day to 100 mg/day, or Any other suitable daily dose may be administered for treatment. Doses can be, for example, 0.01 μg/kg body weight/week to 100 μg/kg body weight/week, or any other suitable dose.

A pharmaceutical composition comprising a selective IL-2Rβγc agonist or IL-2Rβ ligand and/or a compound comprising an IL-2Rγc ligand provided by the present disclosure is administered in the blood or plasma of a subject IL-2Rβ provided by the present disclosure. Selective IL-2Rβγc agonists or compounds, including ligands and/or IL-2Rγc ligands, may be administered to treat cancer in a subject to provide therapeutically effective concentrations. A therapeutically effective concentration of a selective IL-2Rβγc agonist or compound, including an IL-2Rβ ligand and/or an IL-2Rγc ligand provided by the present disclosure, in the blood of a subject is, for example, 0.01 μg/L to 1,000 μg/L. L, 0.1 μg/L to 500 μg/L, 1 μg/L to 250 μg/L, or about 10 μg/L to about 100 μg/L. A therapeutically effective concentration of a selective IL-2Rβγc agonist or compound, including an IL-2Rβ ligand and/or an IL-2Rγc ligand provided by the present disclosure, in the blood of a subject is, for example, at least 0.01 μg/L, at least 0.01 μg/L, It can be 1 μg/L, at least 1 μg/L, at least about 10 μg/L, or at least 100 μg/L. A therapeutically effective concentration of an IL-2Rβ ligand and/or a selective IL-2Rβγc agonist or compound, including an IL-2Rγc ligand, in the blood of a subject is below an amount that causes unacceptable adverse effects, including, for example, adverse effects on homeostasis. could be. A therapeutically effective concentration of an IL-2Rβ ligand and/or a selective IL-2Rβγc agonist or compound, including an IL-2Rγc ligand, in the blood of a subject is an amount sufficient to restore and/or maintain homeostasis in the subject. obtain.

A pharmaceutical composition comprising a selective IL-2Rβγc agonist or a compound comprising an IL-2Rβ ligand and/or an IL-2Rγc ligand is administered to treat a disease of a subject, wherein IL-2Rβ ligand and/or IL-2Rβ ligand and/or IL-2Rγc ligand is present in the blood of the subject. or a therapeutically effective concentration of a selective IL-2Rβγc agonist or compound, including an IL-2Rγc ligand, for example, at least about 4 hours, at least about 6 hours, at least about 12 hours, at least 1 day, at least 2 days, at least 3 days, or can be provided for at least one week.

The amount of IL-2Rβ ligand and/or selective IL-2Rβγc agonist or compound, including IL-2Rγc ligand, administered may vary during the treatment regimen.

Pharmaceutical compositions provided by the present disclosure are selective IL-2Rβγc agonists or compounds comprising IL-2Rβ ligands and/or IL-2Rγc ligands provided by the present disclosure, in addition to one or more pharmaceutically active compounds. can further include compounds such as Such compounds are useful, for example, to treat cancers that have been treated with compounds comprising selective IL-2Rβγc agonists, IL-2Rβ ligands and/or IL-2Rγc ligands, or to treat IL-2Rβ ligands and/or Or selective IL-2Rβ ligands and/or IL-2Rγc ligands to treat diseases, disorders or conditions other than cancer that have been treated with selective IL-2Rβγc agonists or compounds that include IL-2Rγc ligands. To enhance the efficacy of selective IL-2Rβγc agonists or compounds, including IL-2Rβ ligands and/or IL-2Rγc ligands, for treating side effects caused by administering IL-2Rβγc agonists or compounds, and/or to modulate the activity of selective IL-2Rβγc agonists or compounds, including IL-2Rβ ligands and/or IL-2Rγc ligands.

Selective IL-2Rβγc agonists or compounds, including IL-2Rβ ligands and/or IL-2Rγc ligands provided by the present disclosure, can be used in combination with at least one other therapeutic agent. Selective IL-2Rβγc agonists or compounds, including IL-2Rβ ligands and/or IL-2Rγc ligands, may be administered to a patient along with another compound for treating cancer in a subject. In certain embodiments, the at least one other therapeutic agent may be a selective IL-2Rβγc agonist or compound, including different selective IL-2Rβ ligands and/or IL-2Rγc ligands. Selective IL-2Rβγc agonists or compounds, including IL-2Rβ ligands and/or IL-2Rγc ligands and at least one other therapeutic agent, may act additively or, in certain embodiments, synergistically. The at least one additional therapeutic agent may be contained in the same pharmaceutical composition or vehicle comprising the IL-2Rβ ligand and/or the selective IL-2Rβγc agonist or compound, including the IL-2Rγc ligand, or in a separate pharmaceutical composition. or contained in a vehicle. Accordingly, methods provided by the present disclosure provide for the administration of selective IL-2Rβγc agonists or compounds, including IL-2Rβ ligands and/or IL-2Rγc ligands, in addition to cancer or a disease different from cancer, Further comprising administering one or more therapeutic agents effective to treat the disorder or condition. Methods provided by the present disclosure include administration of selective IL-2Rβγc agonists or compounds, including IL-2Rβ ligands and/or IL-2Rγc ligands and one or more other therapeutic agents, but in combination administration does not interfere with the therapeutic efficacy of the IL-2Rβ ligand and/or the selective IL-2Rβγc agonist or compound comprising the IL-2Rγc ligand or produce an adverse combination effect.

A pharmaceutical composition comprising a selective IL-2Rβγc agonist or compound comprising an IL-2Rβ ligand and/or an IL-2Rγc ligand is a selective IL-2Rβγc agonist comprising an IL-2Rβ ligand and/or an IL-2Rγc ligand and/or It may be administered concurrently with administration of another therapeutic agent, which may be part of the same pharmaceutical composition as the compound. Selective IL-2Rβγc agonists and/or compounds, including IL-2Rβ ligands and/or IL-2Rγc ligands, can be administered before or after administration of another therapeutic agent. In certain embodiments of combination therapy, the combination therapy includes selective IL-2Rβ ligands and/or IL-2Rγc ligands, eg, to minimize adverse drug effects associated with certain drugs. It can involve alternating administration between the 2Rβγc agonist or compound and a composition comprising another therapeutic agent. When selective IL-2Rβγc agonists or compounds, including IL-2Rβ ligands and/or IL-2Rγc ligands, are co-administered with another therapeutic agent that can potentially produce adverse drug effects, including, for example, toxicity, other can be administered at doses below the threshold for inducing adverse drug reactions.

Pharmaceutical compositions comprising selective IL-2Rβ and/or IL-2Rγc ligand-comprising IL-2Rβγc agonists or compounds provided by the present disclosure include, for example, IL-2Rβ and/or IL-2Rγc ligand-comprising selective In order to enhance, modulate and/or control the release, bioavailability, therapeutic efficacy, therapeutic potency and/or stability of an IL-2Rβγc agonist or compound, it may be administered with one or more substances. For example, a pharmaceutical composition comprising a selective IL-2Rβγc agonist or compound comprising an IL-2Rβ ligand and/or an IL-2Rγc ligand is a selective IL-2Rβγc agonist or compound comprising an IL-2Rβ ligand and/or an IL-2Rγc ligand. It can be co-administered with active agents that have pharmacological effects that enhance the therapeutic efficacy of the compounds.

A selective IL-2Rβγc agonist, a compound comprising an IL-2Rβ ligand and/or an IL-2Rγc ligand, or a pharmaceutical composition comprising any of the foregoing, comprises a selective IL-2Rβ ligand and/or IL-2Rγc ligand. It may be administered in conjunction with agents known or believed to be effective in treating cancer in a patient, such as the same cancer being treated with a selective IL-2Rβγc agonist or compound, including.

Selective IL-2Rβγc agonists, IL-2Rβ ligands and/or compounds comprising IL-2Rγc ligands, or pharmaceutical compositions comprising any of the foregoing are known to or believed to interfere with proliferation. can be administered in conjunction with an agent prescribed for

In certain embodiments, a selective IL-2Rβγc agonist, a compound comprising an IL-2Rβ ligand and/or an IL-2Rγc ligand, or a pharmaceutical composition comprising any of the foregoing is known to interfere with metabolism. or may be administered in conjunction with agents that are believed to interfere with metabolism. Selective IL-2Rβγc agonists, IL-2Rβ ligands and/or compounds comprising IL-2Rγc ligands, or pharmaceutical compositions comprising any of the foregoing are known to interfere with or are believed to interfere with mitochondrial metabolism. It can be administered in conjunction with possible agents. Selective IL-2Rβγc agonists, IL-2Rβ ligands and/or compounds comprising IL-2Rγc ligands, or pharmaceutical compositions comprising any of the foregoing are known to be anti-metabolites or anti-metabolites. It may also be administered in conjunction with drugs that are believed to be metabolites. Selective IL-2Rβγc agonists, IL-2Rβ ligands and/or compounds comprising IL-2Rγc ligands, or pharmaceutical compositions comprising any of the foregoing, are known to interfere with or are believed to interfere with RNA transcription. It may also be administered in conjunction with possible agents. A selective IL-2Rβγc agonist, a compound comprising an IL-2Rβ ligand and/or an IL-2Rγc ligand, or a pharmaceutical composition comprising a selective IL-2Rβγc agonist is known to interfere with or is believed to interfere with RNA translation. It may also be administered in conjunction with possible agents. Selective IL-2Rβγc agonists, IL-2Rβ ligands and/or compounds comprising IL-2Rγc ligands, or pharmaceutical compositions comprising any of the foregoing, are known to interfere with protein synthesis or are known to interfere with protein synthesis. It can be administered in conjunction with possible agents. A selective IL-2Rβγc agonist, an IL-2Rβ ligand and/or a compound comprising an IL-2Rγc ligand, or a pharmaceutical composition comprising any of the foregoing prevents the synthesis of precursors for DNA synthesis and replication. may be administered in conjunction with agents known to or thought to interfere with A selective IL-2Rβγc agonist, an IL-2Rβ ligand and/or a compound comprising an IL-2Rγc ligand, or a pharmaceutical composition comprising any of the foregoing, is known to interfere with or is believed to interfere with purine synthesis. It can be administered in conjunction with possible agents. A selective IL-2Rβγc agonist, an IL-2Rβ ligand and/or a compound comprising an IL-2Rγc ligand, or a pharmaceutical composition comprising any of the foregoing, is known to interfere with or is believed to interfere with nucleoside synthesis. It can be administered in conjunction with possible agents. A selective IL-2Rβγc agonist, an IL-2Rβ ligand and/or a compound comprising an IL-2Rγc ligand, or a pharmaceutical composition of any of the foregoing is known to interact with mTOR, or is capable of interacting with mTOR. It may also be administered in conjunction with an agent believed to do so. Selective IL-2Rβγc agonists, IL-2Rβ ligands and/or compounds comprising IL-2Rγc ligands, or pharmaceutical compositions of any of the foregoing are known or believed to interact as mTOR inhibitors. It may be administered in conjunction with a drug. Selective IL-2Rβγc agonists, IL-2Rβ ligands and/or compounds comprising IL-2Rγc ligands, or pharmaceutical compositions of any of the foregoing are known or believed to interfere with cell cycle checkpoints. It may be administered in conjunction with a drug.

A compound comprising an IL-2Rβγc agonist, an IL-2Rβ ligand and/or an IL-2Rγc ligand, or a pharmaceutical composition of any of the foregoing, is a CTLA-4 inhibitor such as ipilimumab, a PD1 inhibitor such as pembrolizumab and nivolumab , and PD-LI inhibitors such as atezolizumab, avelumab, and durvalumab. A compound comprising an IL-2Rβγc agonist, an IL-2Rβ ligand and/or an IL-2Rγc ligand, or a pharmaceutical composition of any of the foregoing, is an immunomodulator such as CD137/4-1BB, CD27, GIYR, and/or OC40 may be administered in conjunction with

Selective IL-2Rβγc agonists, IL-2Rβ ligands and/or compounds comprising IL-2Rγc ligands, or pharmaceutical compositions of any of the foregoing, are known to be cytotoxic or It may also be administered in conjunction with drugs that are believed to be sexually active. Selective IL-2Rβγc agonists, IL-2Rβ ligands and/or compounds comprising IL-2Rγc ligands, or pharmaceutical compositions of any of the foregoing, are known to be cytostatic or It may also be co-administered with agents that are believed to be anti-proliferative. Selective IL-2Rβγc agonists, IL-2Rβ ligands and/or compounds comprising IL-2Rγc ligands, or pharmaceutical compositions of any of the foregoing are known or believed to cause DNA damage. It may be administered in conjunction with an agent prescribed for Selective IL-2Rβγc agonists, IL-2Rβ ligands and/or compounds comprising IL-2Rγc ligands, or pharmaceutical compositions of any of the foregoing are known or believed to cause cell cycle arrest. It may be administered in conjunction with a drug. selective IL-2Rβγc agonists, IL-2Rβ ligands and/or compounds comprising IL-2Rγc ligands, or pharmaceutical compositions comprising any of the foregoing are known to cause mitotic cell death, or It may also be administered in conjunction with an agent believed to cause it.

Selective IL-2Rβγc agonists, IL-2Rβ ligands and/or compounds comprising IL-2Rγc ligands, or pharmaceutical compositions of any of the foregoing are known to modulate drug resistance or are believed to modulate drug resistance. It may also be administered in conjunction with possible agents. Selective IL-2Rβγc agonists, IL-2Rβ ligands and/or compounds comprising IL-2Rγc ligands, or pharmaceutical compositions of any of the foregoing are known to reduce multidrug resistance or It may also be co-administered with an agent believed to cause A selective IL-2Rβγc agonist, an IL-2Rβ ligand and/or a compound comprising an IL-2Rγc ligand, or a pharmaceutical composition of any of the foregoing is known to interact with membrane proteins or is believed to interact with membrane proteins. It may also be administered in conjunction with possible agents. Are selective IL-2Rβγc agonists, IL-2Rβ ligands and/or compounds comprising IL-2Rγc ligands, or pharmaceutical compositions comprising any of the foregoing known to interact with plasma membrane proteins? , or in conjunction with drugs with which they are believed to interact. selective IL-2Rβγc agonists, IL-2Rβ ligands and/or compounds comprising IL-2Rγc ligands, or pharmaceutical compositions comprising any of the foregoing are known to interact with nuclear membrane proteins, or Alternatively, it may be administered in conjunction with agents with which it is believed to interact. A selective IL-2Rβγc agonist, a compound comprising an IL-2Rβ ligand and/or an IL-2Rγc ligand, or a pharmaceutical composition comprising any of the foregoing can interact with the major vault protein(s). It may also be administered in conjunction with agents known to or suspected of interacting with them. A selective IL-2Rβγc agonist, a compound comprising an IL-2Rβ ligand and/or an IL-2Rγc ligand, or a pharmaceutical composition comprising any of the foregoing interacts with the product of the MVP (major vault protein) gene. It may also be administered in conjunction with drugs known to interact with the drug.

Selective IL-2Rβγc agonists, IL-2Rβ ligands and/or compounds comprising IL-2Rγc ligands, or pharmaceutical compositions of any of the foregoing are known to modulate glutathione levels or will modulate glutathione levels. It may also be administered in conjunction with possible agents. Are selective IL-2Rβγc agonists, IL-2Rβ ligands and/or compounds comprising IL-2Rγc ligands, or pharmaceutical compositions comprising any of the foregoing known to modulate intracellular glutathione levels? , or in conjunction with an agent that is believed to modulate. Compounds comprising selective IL-2Rβγc agonists, IL-2Rβ ligands and/or IL-2Rγc ligands, or pharmaceutical compositions comprising selective IL-2Rβγc agonists, or compounds comprising IL-2Rβ ligands and/or IL-2Rγc ligands may be administered in conjunction with agents known or believed to lower intracellular glutathione levels. Compounds comprising selective IL-2Rβγc agonists, IL-2Rβ ligands and/or IL-2Rγc ligands, or pharmaceutical compositions comprising selective IL-2Rβγc agonists, or compounds comprising IL-2Rβ ligands and/or IL-2Rγc ligands may be administered in conjunction with agents known or believed to reduce glutathione uptake into cells. Selective IL-2Rβγc agonists, IL-2Rβ ligands and/or compounds comprising IL-2Rγc ligands, or pharmaceutical compositions comprising any of the foregoing, are known to reduce glutathione synthesis or reduce glutathione synthesis. It may also be administered in conjunction with an agent believed to do so. Are selective IL-2Rβγc agonists, IL-2Rβ ligands and/or compounds comprising IL-2Rγc ligands, or pharmaceutical compositions comprising any of the foregoing known to reduce intracellular glutathione synthesis? , or in conjunction with agents thought to reduce it.

Selective IL-2Rβγc agonists, IL-2Rβ ligands and/or compounds comprising IL-2Rγc ligands, or pharmaceutical compositions comprising any of the foregoing are known to or are believed to interfere with angiogenesis. It may also be administered in conjunction with possible agents. Selective IL-2Rβγc agonists, IL-2Rβ ligands and/or compounds comprising IL-2Rγc ligands, or pharmaceutical compositions comprising any of the foregoing, are known to reduce angiogenesis or reduce angiogenesis. It can be administered in conjunction with agents believed to do so. Selective IL-2Rβγc agonists, IL-2Rβ ligands and/or compounds comprising IL-2Rγc ligands, or pharmaceutical compositions comprising any of the foregoing, are known to promote angiogenesis or promote angiogenesis. It may also be administered in conjunction with an agent believed to do so.

Selective IL-2Rβγc agonists, IL-2Rβ ligands and/or compounds comprising IL-2Rγc ligands, or pharmaceutical compositions comprising any of the foregoing, are known to interfere with hormone homeostasis or are believed to interfere with hormone homeostasis. It can be administered in conjunction with possible agents. In certain embodiments, a selective IL-2Rβγc agonist, a compound comprising an IL-2Rβ ligand and/or an IL-2Rγc ligand, or a pharmaceutical composition comprising any of the foregoing is known to interfere with hormone synthesis. or may be administered in conjunction with agents that are believed to interfere. A selective IL-2Rβγc agonist, a compound comprising an IL-2Rβ ligand and/or an IL-2Rβ ligand and/or an IL-2Rγc ligand, or a pharmaceutical composition comprising any of the foregoing prevents hormone receptor binding. may be administered in conjunction with agents known to or thought to interfere with Selective IL-2Rβγc agonists, IL-2Rβ ligands and/or compounds comprising IL-2Rγc ligands, or pharmaceutical compositions comprising any of the foregoing are known to interfere with or interfere with hormone signaling. It can be administered in conjunction with an agent that is believed to be

selective IL-2Rβγc agonists, IL-2Rβ ligands and/or compounds comprising IL-2Rγc ligands, or pharmaceutical compositions comprising any of the foregoing are known to interfere with growth factor homeostasis, or It may also be administered in conjunction with drugs that are believed to interfere. Selective IL-2Rβγc agonists, IL-2Rβ ligands and/or compounds comprising IL-2Rγc ligands, or pharmaceutical compositions comprising any of the foregoing are known to interfere with or interfere with growth factor synthesis. It can be administered in conjunction with an agent that is believed to be selective IL-2Rβγc agonists, IL-2Rβ ligands and/or compounds comprising IL-2Rγc ligands, or pharmaceutical compositions comprising any of the foregoing are known to interfere with growth factor receptor expression, or Or it can be administered in conjunction with drugs that are thought to interfere. Selective IL-2Rβγc agonists, compounds comprising IL-2Rβ ligands and/or IL-2Rγc ligands, or pharmaceutical compositions comprising any of the foregoing are known to interfere with growth factor binding to growth factor receptors. It may be administered in conjunction with drugs that are considered to interfere with or are thought to interfere. Selective IL-2Rβγc agonists, compounds comprising IL-2Rβ ligands and/or IL-2Rγc ligands, or pharmaceutical compositions comprising any of the foregoing are known to interfere with growth factor binding to growth factor receptors. It may be administered in conjunction with drugs that are considered to interfere with or are thought to interfere. Are selective IL-2Rβγc agonists, IL-2Rβ ligands and/or compounds comprising IL-2Rγc ligands, or pharmaceutical compositions comprising any of the foregoing known to interfere with growth factor receptor signaling? , or in conjunction with drugs that are thought to interfere. Selective IL-2Rβγc agonists, compounds comprising IL-2Rβ ligands and/or IL-2Rγc ligands, or pharmaceutical compositions comprising any of the foregoing are known to interfere with Hedgehog (Hh) signaling. or may be administered in conjunction with drugs that are believed to interfere. Compounds comprising selective IL-2Rβγc agonists, IL-2Rβ ligands and/or IL-2Rγc ligands, or pharmaceutical compositions comprising any of the foregoing are known to inhibit hedgehog pathway signaling. , or in conjunction with an agent that is believed to inhibit. A selective IL-2Rβγc agonist, a compound comprising an IL-2Rβ ligand and/or an IL-2Rγc ligand, or a pharmaceutical composition comprising any of the foregoing inhibits ALK (aplastic lymphoma kinase) pathway signaling. may be administered in conjunction with agents known or believed to inhibit Selective IL-2Rβγc agonists, compounds comprising IL-2Rβ ligands and/or IL-2Rγc ligands, or pharmaceutical compositions comprising any of the foregoing are known to inhibit non-homologous end joining (NHEJ). or may be administered in conjunction with agents that are believed to inhibit it.

Selective IL-2Rβγc agonists, IL-2Rβ ligands and/or compounds comprising IL-2Rγc ligands, or pharmaceutical compositions of any of the foregoing, are VEGFR (vascular endothelial growth factor receptor) inhibitors, RTK (receptor tyrosine kinase) inhibitor, sodium channel current blocker, aFAK (focal adhesion kinase) inhibitor, GLI (glioma-associated oncogene) inhibitor, GLI1 inhibitor, GLI2 inhibitor, GLI3 inhibitor, MAPK (mitogen activated protein kinase) inhibitor, MAPK/ERK pathway (also known as Ras-Raf-MEK-ERK pathway) inhibitor, MEK1 inhibitor, MEK2 inhibitor, MEK5 inhibitor, MEK5/ERK5 inhibitor, RTA (renal tubular acidosis) inhibitor, ALK (aplastic lymphoma kinase) inhibitor, Aa LK kinase inhibitor, nuclear translocation inhibitor, PORCN (porcupin) inhibitor, 5-ARI (5α reductase inhibitor), topoisomerase inhibitor agents, Ras (rat sarcoma) inhibitors, K-ras inhibitors, CERK (ceramide kinase) inhibitors, PKB (protein kinase B aka AKT) inhibitors, AKT1 inhibitors, EZH2 (enhancer of zest homolog 2) inhibitors , Bet (bromodomain and extraterminal domain motifs) inhibitors, SYK (apreentyrosine kinase) inhibitors, JAK (Janas kinase) inhibitors, SYK/JAK inhibitors, IDO (indoleamine-pyrrole 2,3-dioxygenase ) inhibitor, IDO1 inhibitor, RXR (retinoic acid X receptor) activator, selective RXR activator, p-glycoprotein inhibitor, ERK inhibitor, PI3K (phosphatidylinositol-4,5-bisphosphate 3-kinase) inhibitor, BRD (bromodomain containing protein) inhibitor, BRD2 inhibitor, BRD3 inhibitor, BRD4 inhibitor, BRDT (bromodomain testis specific protein) inhibitor, reverse transcriptase inhibitor, NRT (nucleoside analog reverse transcriptase) inhibitors, PIM (preliminary integration of Moloney virus) inhibitors, EGFR (epidermal growth factor receptor) inhibitors, photosensitizers, radiosensitizers, ROS (proto-oncogene, receptor tyrosine kinase inhibitors such as somatic tyrosine kinase) inhibitors, ROS1 (proto-oncogene 1) inhibitors, CK (casein kinase) inhibitors, CK2 inhibitors, Bcr-Abl (cluster region-Abelson proto-oncogene) dasatinib , microtubule stabilizing agents, microtubule depolymerization/disassembly inhibitors, DNA intercalators, androgen receptor antagonists, chemoprotective agents, HDAC (histone deacetylase) inhibitors, DPP (dipeptidyl peptidase) inhibitors, DPP- 4 inhibitors, BTK (Bruton's tyrosine kinase) inhibitors, kinase inhibitors such as imatinib, tyrosine kinase inhibitors such as nilotinib, ARP (poly (ADP-ribose) polymerase) inhibitors, CDK (cyclin dependent kinase) inhibitors agents, CDK4 inhibitors, CDK6 inhibitors, CDK4/6 inhibitors, HIF1α (hypoxia-inducible factor 1-α) inhibitors, DNA ligase inhibitors, DNA ligase IV inhibitors, NHEJ (non-homologous end joining inhibitors), DNA ligase IV, NHEJ inhibitors and RAF inhibitors, TKI and RAF inhibitors, TKI and RAF inhibitors such as sorafenib, PDT (photodynamic therapy) sensitizers, ATR (Ataxia telangiectasia and Rad3-related protein kinase) inhibitors, or combinations of any of the above

A compound comprising a selective IL-2Rβγc agonist, an IL-2Rβ ligand and/or an IL-2Rγc ligand, or a pharmaceutical composition of any of the foregoing is a VEGFR inhibitor such as, for example, fluquintinib, motesanib/AMG-706, vatalanib, RTK inhibitors such as ponatinib, sodium channel blockers such as GS967, FAK inhibitors such as TAE226, GLI1 and GLI2 inhibitors such as GANT61, MEK inhibitors such as binimetinib, RTA inhibitors such as linifanib, ALK inhibitors such as brigustinib agents, DNA alkylating agents such as bromopyruvate and thiotepa, nuclear translocation factors such as JSH-23, PORCn inhibitors such as Wnt-C59, 5α reductase inhibitors such as dutasteride, topoisomerase inhibitors such as carbicin, Kobe0065 RAS inhibitors such as NVP-231, CerK inhibitors such as NVP-231, AKT inhibitors such as uprosertib, EZH2 inhibitors such as GSK-503, BET bromodomain inhibitors such as OTX015, MEK5/ERK5 inhibitors such as BIX02189, selduratinib Syl/JAK inhibitors such as NLG919, IDO 1 inhibitors such as NLG919, retinoic acid X receptor activators such as bexrotene, PGP inhibitors such as acotiamide or actiamide HCl, Erk inhibitors such as SCH772984, PI3K inhibitors such as gedatricib. agents, JAK inhibitors such as ruxolitinib, AKT inhibitors such as afresertib or afresertib HCl, ALK1 inhibitors such as ceritinib, HDAC inhibitors such as abexinostat, DPP inhibitors such as oamarigliptin, EGFR inhibitors such as gefitinib agents, EZH2 inhibitors such as GSK126, BTK inhibitors such as ibrutinib, kinase inhibitors such as imatinin HCl, IDO inhibitors such as INCB024360, DNA cross-linkers such as mitomycin C, tyrosine kinase inhibitors such as nilotinib, olaparib and the like. PARP inhibitors, tubilin stabilizing enhancers such as paclitaxel, CDK4/6 inhibitors such as palbociclib, RTK inhibitors such as sunitinib, PDT sensitizers such as tslsporfin, p-glycoprotein inhibitors such as tariquidar, VE-822 HDAC inhibitors such as PCI-24781, DPP inhibitors such as omarigliptin, EGFR inhibitors such as gefitinib, EZH2 inhibitors such as GSK126, BTK inhibitors such as irbutinib, IDO inhibitors such as INCB024360, Or it can be administered in combination with any of the above.

For example, a selective IL-2Rβγc agonist, an IL-2Rβ ligand and/or a compound comprising an IL-2Rγc ligand, or any of the foregoing may be combined with another chemotherapeutic agent such as N-acetylcysteine (NAC), adriamycin, alemtuzumab , amifostine, arsenic trioxide, ascorbic acid, bendamustine, bevacizumab, bortezomib, busulfan, buthionine sulfoxime, carfilzomib, carmustine, clofarabine, cyclophosphamide, cyclosporine, cytarabine, dasatinib, datinomycin, defibrotide, dexamethaxone, docetaxel, doxorubicin, Etoposide, Filgrastim, Floxalidine, Fludarabine, Gemcitabine, Interferon-α, Ipilimumab, Lenalidomide, Leucovorin, Melphalan, Mycophenolate Mofetil, Paclitaxel, Palifermin, Panobinostat, Pegfilastim, Prednisolone, Prednisone, Revlimid, Rituximab, Sirolimus, 2- It may be administered in combination with sodium mercaptoethanesulfonate (MESNA), sodium thiosulfate, tacrolimus, temozolomide, thalidomide, thioguanine, thiotepa, topotecan, Velcade, or any combination of the foregoing. In certain embodiments, selective IL-2Rβγc agonists and/or pharmaceutical compositions thereof comprise one or more antimetabolites such as folic acid analogs, pyrimidine analogs such as fluorouracil, floxalysine, and cytosine arabinoside; Purine analogues such as mercaptopurine, thiognaine, and pentostatin; natural products such as vinblastine, vincristine, etoposide, tertiposide, dactinomycin, daunorubicin, doxarubicin, bleomycin, mitamycin, mitomycin C, L-asparaginase, and interferon alpha; - platinum and platinum coordination complexes such as carboplatin, mitoxantrone, hydroxyurea, procarbazine, prednisone, hydroxyprogesterone caproate, medroxyprogesterone acetate, medroxyprogesterone acetate, megestrol acetate, diethylstilbestrol hormones and antagonists such as , ethinylestradiol, tamoxifen, testosterone propionate, fluoxymesterone, flutamide, and leuprolide; antiangiogenic agents or inhibitors such as angiostatin, retinoic acid, paclitaxel, estradiol derivatives, and thiazolopyrimidine derivatives; , anti-apoptotic agents, triptolide, colchicine, luliconazole, radiotherapy.

Selective IL-2Rβγc agonists, or compounds containing IL-2Rβ ligands and/or IL-2Rγc ligands, may be co-administered with compounds that inhibit DNA repair, such as O6-benzylguanine (O6-BG).

A selective IL-2Rβγc agonist, an IL-2Rβ ligand and/or a compound comprising an IL-2Rγc ligand, or a pharmaceutical composition comprising any of the foregoing may be combined with one or more chemotherapeutic agents such as abarelix, abiraterone, acetic acid Abiraterone, n-acetylcysteine, aclarubicin hydrochloride, adriamycin, adenine, afatinib, afatinib dimaleate, alemtuzumab, alendronate sodium, alitretinoin, allopurinol sodium, altretamine, amifostine, aminoglutethimide, aminolevulinic acid, amrubicin, amsacrine, ana strozol, angiostatin, apremilast, aprepitant, arsenic trioxide, ascorbic acid, l-asparaginase, azacitidine, azathioprine sodium, bazedoxifene (serum), belinostat, bendamustine hcl, o6-benzylguanine (o6-bg), bevacizumab, bexarotene, bicalutamide, viricodar, bleomycin sulfate, bortezomib, bosutinib, brivudine, buserelin, busulfan, buthionine sulfoxime, cabazitaxel, cabozantinib, capecitabine, carboplatin, carboquone, carfilzomib, carmoflu, carmustine, ceritinib, chlorambucil, cisplatin, cladribine, clodronate disodium , clofarabine, crizotinib, cyclophosphamide, cyclosporine, cytarabine, cytosine arabinoside, dabrafenib, dacarbazine, dactinomycin, dasatinib, datinomycin, daunorubicin, decitabine, defribrotide, degarelix acetate, dexamethasone, dexrazoxane hydrochloride, diaziquone, diethylstilbest rol, docetaxel, doxfluridine, doxorubicin hydrochloride, doxorubicin free base, drostanolone propionate, dutasteride, eltrombopag, enzalutamide, epirubicin hydrochloride, eribulin mesylate, erlotinib hydrochloride, estramustine sodium phosphate, ethinyl estradiol, etoposide phosphate, etoposide , everolimus, exemestane, fentanyl, filgrastim, fingolimod, floxuridine, fludarabine phosphate, fluorouracil, fluoxymesterone, flutamide, formestane, formylmelphalan, fosaprepitant, fotemustine, fulvestrant, gefitinib, gemcitabine hydrochloride , gemcitabine free base, glutathione, glyciphosphoramide, glyphosphine, goserelin acetate, granisetron hydrochloride, heptaplatin, hexyl 5-aminolevulinate, histrelin acetate, hydroxyprogesterone caproate, hydroxyurea, ibandronate sodium, ibrutinib, icotinib , idarubicin HCl, idelalisib, idoxuridine, ifosfamide, interferon-alpha, imatinib mesylate, imiquimod, ingenol mebutate, ipilimumab, irinotecan hydrochloride, ixabepilone, lanreotide acetate, lapatinib free base, lapatinib ditosylate, lasofoxifen, lenalidomide , letrozole, leucovorin calcium, leuprolide acetate, levamisole hydrochloride, levofolinate calcium, iobenguane, lobaplatin, lomustine, maropitant, masoprocol, mechlorethamine hydrochloride, megestrol acetate, medroxyprogesterone acetate, melphalan hydrochloride, mercaptopurine, mercaptoethanesulfonic acid sodium, methotrexate, methoxsalen, methyl aminolevulinate, methylene blue, methylisoindigotin, mifamurtide, miltefosine, miriplatin, mitamycin, mitobronitol, mitomycin C, mitotane, mitoxantrone hydrochloride, mycophenolate mofetil, naviximols, nafarelin, nandrolone, nedaplatin, Nelarabine, netupitant, nilotinib, nilutamide, nimustine, nintedanib, nocodazole, octreotide, olaparib, omacetaxine mepesuccinate, ondansetron hydrochloride, oxaliplatin, paclitaxel, palbociclib, palyfermin, palonosetron hydrochloride, pamidronate disodium, panobinostat, pasireotide , pazopanib hydrochloride, pegfilastim, pemetrexed disodium, pentostatin, peplomycin, pipobroman, pirarubicin, plelixafor, plicamycin, pomalidomide, ponatinib, porfimer sodium, porphyromycin, pralatrexate, prednimustine, prednisolone, prednisone, procarbazine hydrochloride , quinagolide hydrochloride, raloxifene, raltitrexed, radtinib, ranimustine, retinoic acid, levlimid, rituxinab, romidepsin, ruxolitinib, ruxolitinib phosphate, semustine, sirolimus, sodium thiosulfate, sorafenib free base, sorafenib tosylate, streptozocin, sufentanil, sunitinib , tacrolimus, talaporfin sodium, tamivalotene, tamoxifen citrate, tapentadol, temoporfin, temozolomide, temsirolimus, teniposide, teriflunomide, tertiposide, testolactone, testosterone propionate, thalidomide, thioguanine, thiotepa, thymalfasin, toceranib phosphate, topotecan hydrochloride, with toremifene citrate, trabectedine, trametinib, tretinoin, trilostane, triptorelin, tropisetron, uramustine, valrubicin, vandetanib, vedotin, vemurafenib, verteporfin, vinblastine, vincristine sulfate, vincristine free base, vindesine, vinorelbine tartrate, vorinostat, and zoledronic acid Can be administered in combination.

Selective IL-2Rβγc agonists, IL-2Rβ ligands and/or compounds comprising IL-2Rγc ligands, or pharmaceutical compositions comprising any of the foregoing are abemaciclib, abiraterone acetate, ABVD, ABVE, ABVE-PC, AC, acalabrutinib , AC-T, ADE, adtrastuzumab emtansine, afatinib dimaleate, aldesleukin, alectinib, alemtuzumab, alpelisib, amifostine, aminolevulinic acid hydrochloride, anastrozole, apalutamide, aprepitant, arsenic trioxide, asparaginase erwinia chrysanthemi, atezolizumab, avelumab, axicabutagene siloleucel, axitinib, azacitidine, BEACOPP, belinostat, bendamustine hydrochloride, BEP, bevacizumab, bexarotene, bicalutamide, binimetinib, bleomycin sulfate, blinatumomab, bortezomib, bosutinib, brentuximab vedotin, brigatinib, BuMel , busulfan, cabazitaxel, cabozantinib-s-malate, CAF, calaspargase pegol-mknl, capecitabine, capracitabine-yhdp, CAPOX, carboplatin, carboplatin-taxol, carfilzomib, carmustine, carmustine implant, CEM, semiplimab-rwlc, ceritinib, cetuximab, CEV, chlorambucil, chlorambucil-prednisone, CHOP, cisplatin, cladribine, clofarabine, CMF, cobimetinib, copanlisib hydrochloride, COPDAC, COPP, COPP-ABV, crizotinib, CVP, cyclophosphamide, cytarabine, cytarabine liposomes, mesyl dabrafenib acid, dacarbazine, dacomitinib, dactinomycin, daratumumab, darbepoetin alfa, dasatinib, daunorubicin hydrochloride, daunorubicin hydrochloride and cytarabine liposomes, decitabine, defibrotide sodium, degarelix, denileukin-diffitox, denosumab, dexamethasone, dexrazoxane hydrochloride, dinutuximab , docetaxel, doxorubicin hydrochloride, doxorubicin hydrochloride liposomes, durvalumab, duvelisib, elotuzumab, ertrombopaguolamine, emapalumab-lzsg, enacidenib mesylate, encorafenib, enzalutamide, epirubicin hydrochloride, EPOCH, epoetin alfa, erdafitinib, eribulin mesylate, erlotinib hydrochloride , etoposide, etoposide phosphate, everolimus, exemestane, fec, filgrastim, fludarabine phosphate, fluorouracil injection, fluorouracil-topical, flutamide, forfylli, forfylli-bevacizumab, forfylli-cetuximab, forfylinox, forfox, fostamatinib disodium , FU-LV, fulvestrant, gefitinib, gemcitabine hydrochloride, gemcitabine-cisplatin, gemcitabine-oxaliplatin, gemtuzumab ozogamicin, gilteritinib fumarate, glasdegib maleate, glucarpidase, goserelin acetate, granisetron, HPV bivalent vaccine , HPV bivalent vaccine, recombinant HPV nonvalent vaccine, HPV nonvalent vaccine, HPV nonvalent vaccine recombinant, HPV tetravalent vaccine, HPV uadrivalent vaccine recombinant, hydroxyurea, hyper-CVAD, ibrituzumab tiuxetan , ibrutinib, ICE, idarubicin hydrochloride, idelalisib, ifosfamide, imatinib mesylate, imiquimod, inotuzumab ozogamicin, interferon alpha-2b recombinant, iobenguane I ¹³¹ , ipilimumab, irinotecan hydrochloride, irinotecan hydrochloride liposomes, ivosidenib, ixabepilone , ixazomib citrate, JEB, lanreotide acetate, lapatinib ditosylate, larotrectinib sulfate, lenalidomide, lenvatinib mesylate, letrozole, leucovorin calcium, leuprolide acetate, lomustine, loratinib, lutetium Lu 177-dotatate, mechlorethamine hydrochloride, megestrol acetate, melphalan, melphalan hydrochloride, mercaptopurine, mesna, methotrexate, methylnaltrexone bromide, midostaurin, mitomycin c, mitoxantrone hydrochloride, mogamulizumab-kpkc, moxetumomab pasudotox-tdfk, MVAC, necitumumab, nerarabine, maleic acid neratinib, netupitant and palonosetron hydrochloride, nilotinib, nilutamide, tosylate niraparib monohydrate, nivolumab, obinutuzumab, OEPA, ofatumumab, OFF, olaparib, olalatumab, omacetaxine mepesuccinate, ondansetron hydrochloride, OPPA, osimertinib mesylate, oxaliplatin, paclitaxel, paclitaxel albumin-stabilized nanoparticles, PAD, palbociclib, palifermin, palonosetron hydrochloride, palonosetron hydrochloride and netupitant, pamidronate disodium, panitumumab, panobinostat, pazopanib hydrochloride, PCV, PEB, peguasparagase , pegfilgrastim, peginterferon alfa-2b, pembrolizumab, pemetrexed disodium, pertuzumab, plelixafor, polatuzumab vedotin-piiq, pomalidomide, ponatinib hydrochloride, pralatrexate, prednisone, procarbazine hydrochloride, propranolol hydrochloride, Radium 223 chloride, raloxifene hydrochloride, ramucirumab, rasburicase, rablizumab-cwvz, R-CHOP, R-CVP, recombinant HPV bivalent vaccine, recombinant HPV nonvalent vaccine, recombinant HPV tetravalent vaccine, recombinant interferon alpha-2b , regorafenib, R-EPOCH, ribociclib, R-ICE, rituximab, rituximab and hyaluronidase human, lorafenib hydrochloride, romidepsin, romiplostim, rucaparib camsylate, ruxolitinib phosphate, siltuximab, sipuleucel-t, sonitinib, sorafenib tosylate, Stanford V, sunitinib malate, TAC, tagraxofusp-erzs, talazoparbutosylate, talc, tarimogenlaherparepvec, tamoxifen citrate, temozolomide, temsirolimus, thalidomide, thioguanine, thiotepa, tisagenleklucel, tocilizumab, hydrochloride Topotecan, toremifene, TPF, trabectedin, trametinib, trastuzumab, trastuzumab and hyaluronidase-oysk, trifluridine and tipiracil hydrochloride, uridine triacetate, VAC, valrubicin, VAMP, vandetanib, VeIP, vemurafenib, venetoclax, vinblastine sulfate, vincristine sulfate liposomes, tartaric acid It can be administered in combination with one or more chemotherapeutic agents, including vinorelbine, vip, vismodegib, vorinostat, XELIRI, XELOX, Ziv-aflibercept, zoledronic acid, and combinations of any of the above.

The efficacy of administering selective IL-2Rβγc agonist compounds provided by the present disclosure or comprising IL-2Rβ ligands and/or IL-2Rγc ligands to treat cancer has been determined using in vitro and animal studies. as well as in clinical trials.

Selective IL-2Rβγc agonists, compounds comprising IL-2Rβ ligands and/or IL-2Rγc ligands provided by the present disclosure, and/or pharmaceutical compositions of any of the foregoing, in the treatment of cancers listed above can be determined by methods described in the art. For example, screens developed to demonstrate antitumor activity of oncolytic agents are known (Miller, et al., J Med Chem, 1977, 20(3), 409-413; Sweeney, et al., Cancer Res, 1978, 38(9), 2886-2891 and Weiss and Von Hoff, Semin Oncol, 1985, 12 (3Suppl 4), 69-74). Therefore, assaying and using compounds and/or pharmaceutical compositions thereof to treat the diseases or disorders described above is well within the capabilities of one of ordinary skill in the art.

Compounds provided by the present disclosure may be useful in treating autoimmune diseases. Autoimmune diseases are defined as human diseases in which the immune system attacks its own proteins, cells and tissues. A comprehensive list and review of autoimmune diseases can be found in The Autoimmune Diseases (Rose and Mackay, 2014, Academic Press).

IL-2Rβγc agonists, IL-2Rβ ligands and/or compounds comprising IL-2Rγc ligands provided by the present disclosure, and pharmaceutical compositions of any of the foregoing are used to treat inflammatory or autoimmune diseases. can be administered to the patient at

Examples of inflammatory diseases include allergies, Alzheimer's disease, anemia, ankylosing spondylitis, arthritis, atherosclerosis, asthma, autism, arthritis, carpal tunnel syndrome, celiac disease, colitis, Crohn's disease, Congestive heart failure, dermatitis, diabetes, diverticulitis, eczema, fibromyalgia, fibrosis, gallbladder disease, gastroesophageal reflux disease, Hashimoto thyroiditis, heart attack, hepatitis, irritable bowel syndrome, renal failure, lupus, multiple occurrence Sclerosis, nephritis, neuropathy, pancreatitis, Parkinson's disease, psoriasis, polymyalgia, rheumatism, rheumatoid arthritis, sclerosing dermatitis, stroke, surgical complications, and ulcerative colitis.

Examples of autoimmune diseases include Addison's disease, agamaglobulinemia, alopecia, amyloidosis, ankylosing spondylitis, anti-GBM/anti-TBN nephritis, antiphospholipid syndrome, autoimmune angioedema, autoimmune deficiency, autoimmune Immune encephalitis, autoimmune hepatitis, autoimmune inner ear disease, autoimmune myocarditis, autoimmune pancreatitis, autoimmune retinopathy, autoimmune urticaria, axonal neuropathy, Barro's disease, Behcet's disease, benign mucous membrane pemphigus, pemphigus major, Castleman's disease, celiac disease, Chagas disease, chronic inflammatory demyelinating polyneuropathy, chronic inflammatory demyelinating polymyelitis, Churg-Strauss, cicatricial pemphigoid, Cogan's syndrome, cold agglutinin disease, congenital heart block, cocaxily myocarditis, Crest syndrome, Crohn's disease, dermatitis herpetiformis, dermatomyositis, Devick's disease, discoid lupus erythematosus, Dressler's syndrome, endometriosis, eosinophilia Bulbar esophagitis, eosinophilic fasciitis, erythema nodosum, mixed cryoglobulinemia essential, Evans syndrome, fibromyalgia, fibrosing alveolitis, giant cell arteritis, giant cell myocardium glomerulonephritis, Goodpasture's syndrome, granulomatosis with polyangiitis, Graves' disease, Grain-Barré syndrome, Hashimoto's thyroiditis, hemolytic anemia, Henosh-Sonlein purpura, herpes pregnancy or pemphigus pregnancy , hypoammaglobulinemia, IgA nephropathy, IgG4-associated sclerosis, immune thrombocytopenic purpura, myositis inclusive, interstitial cystitis, juvenile arthritis, juvenile diabetes, juvenile myositis, Kawasaki disease, Lambert Eaton's syndrome, leukocytoclastic vasculitis, lichen planus, lichen sclerosus, xylem conjunctivitis, linear IgA disease, lupus, chronic Lyme disease, Meniere's disease, microscopic polyangiitis, mixed connective tissue disease, Mullen ulcer , Mucha-Habermann disease, multiple sclerosis, myasthenia gravis, myositis, narcolepsy, neuromyelitis optica, optic neuritis, neutropenia, ocular cicatricial pemphigoid, optic neuritis, palindromic rheumatoid arthritis, PANDAS , paraneoplastic cerebellar degeneration, paroxysmal nocturnal hemoglobinuria, Parry-Romberg syndrome, squamous epitheliitis, parsonage-Turner syndrome, penfig, peripheral neuropathy, perivenous encephalomyelitis, pernicious anemia, POEMS syndrome, polyarteriolar nodosa inflammation, polygonad syndrome, rheumatic myalgia, polymyositis, post-myocardial infarction syndrome, post-pericardiotomy syndrome, primary biliary cirrhosis, primary sclerosing cholangitis, progesterone dermatitis, psoriasis, psoriatic arthritis, pure erythropoiesis, ganglenosum pyoderma, Raynaud's phenomenon, reactive arthritis, reflex sympathetic dystrophy, relapsing polychondritis, restless leg syndrome, retroperitoneal fibrosis, rheumatic fever, rheumatoid arthritis, sarcoma , Schmidt syndrome, sclerosis, sclerosing dermatitis, subacute bacterial endocarditis, Susak syndrome, sympathetic ophthalmia, Takayasu arteritis, temporal arteritis, thrombocytopenic purpura, Tolosahunt syndrome, transverse Myelitis, type 1 diabetes, ulcerative colitis, undifferentiated connective tissue disease, uveitis, vasculitis, vitiligo, Wegener's granulomatosis.

A selective IL-2Rαβγc agonist, a compound comprising an IL-2Rβ ligand and/or an IL-2Rγc ligand, or a pharmaceutical composition comprising any of the foregoing, corticosteroids such as, for example, prednisone, budesonide, and prednisolone; Janus kinase inhibitors such as tofacitinib; calcineurin inhibitors such as cyclosporine and tacrolimus; mTOR inhibitors such as sirolimus and everolimus; IMDH inhibitors such as azathioprine, leflunomide, and mycophenolate; One or more immunosuppressants, including biologics such as golimumab, infliximab, ixekizumab, natalizumab, rituximab, sequinukinumab, tocilizumab, ustekinuscumab, and vedolizumab; and monoclonal antibodies such as basilixime and clidazumab may be administered in conjunction with

IL-2Rβγc agonists, compounds comprising IL-2Rβ and/or IL-2Rγc ligands provided by the present disclosure, and pharmaceutical compositions of any of the foregoing are administered to a patient to promote T cell activation, proliferation, and metabolism. , and/or differentiation-related diseases may be treated.

IL-2Rβγc agonists, compounds comprising IL-2Rβ and/or IL-2Rγc ligands provided by the present disclosure, and pharmaceutical compositions of any of the foregoing can be administered to a patient to treat organ transplantation.

IL-2Rβγc agonists, compounds comprising IL-2Rβ and/or IL-2Rγc ligands provided by the present disclosure, and pharmaceutical compositions of any of the foregoing are used for treating inflammatory or autoimmune diseases in a subject. It may be administered to the patient along with another compound. The at least one other therapeutic agent can be a different IL-2Rβγc agonist or compound, including IL-2Rβ and/or IL-2Rγc ligands provided by this disclosure. Compounds comprising an IL-2Rβγc agonist, or IL-2Rβ and/or IL-2Rγc ligand, and at least one other therapeutic agent may act additively or synergistically. The at least one additional therapeutic agent can be included in the same pharmaceutical composition or vehicle with the IL-2Rβγc agonist or compound, including IL-2Rβ and/or IL-2Rγc ligands, or in a separate pharmaceutical composition or vehicle. can be included. Accordingly, methods provided by the present disclosure include administering IL-2Rβγc agonists or compounds, including IL-2Rβ and/or IL-2Rγc ligands, to inflammatory or autoimmune diseases, or inflammatory diseases or It further comprises administering one or more therapeutic agents effective to treat diseases, disorders or conditions other than autoimmune diseases. Methods provided by the present disclosure include administration of IL-2Rβγc agonists or compounds, including IL-2Rβ and/or IL-2Rγc ligands and one or more other therapeutic agents, provided that the co-administration is IL-2Rβ and/or do not interfere with the therapeutic efficacy of the IL-2Rβγc agonist or compound, including the IL-2Rγc ligand, or produce adverse combined effects.

A pharmaceutical composition comprising an IL-2Rβγc agonist may be part of the same pharmaceutical composition as a compound comprising an IL-2Rβ and/or IL-2Rγc ligand and comprising an IL-2Rβ and/or IL-2Rγc ligand. It may be administered concurrently with administration of another therapeutic agent, which may be of a different pharmaceutical composition than the IL-2Rβγc agonist or compound. IL-2Rβγc agonists or compounds, including IL-2Rβ and/or IL-2Rγc ligands, can be administered before or after administration of another therapeutic agent. In combination therapy, the combination therapy is an IL-2Rβγc agonist or compound, including an IL-2Rβ and/or IL-2Rγc ligand, and another drug, e.g., to minimize adverse drug effects associated with a particular drug. Alternate administration between compositions containing the therapeutic agent may be included. Other therapeutic agents when IL-2Rβ and/or IL-2Rβγc agonists or compounds, including IL-2Rβ and/or IL-2Rγc ligands, are co-administered with another therapeutic agent that can potentially result in adverse drug effects, including, for example, toxicity can be administered at doses below the threshold for inducing adverse drug reactions.

Pharmaceutical compositions comprising IL-2Rβ and/or IL-2Rγc agonists or compounds, including IL-2Rβ and/or IL-2Rγc ligands, are compounds that are IL-2Rβγc agonists or IL-2Rβ and/or IL-2Rγc and/or IL-2Rβ and/or It can be administered with one or more substances to enhance, modulate and/or control the release, bioavailability, therapeutic efficacy, therapeutic potency, stability, etc. of compounds including IL-2Rγc ligands. compounds comprising IL-2Rβγc agonists or IL-2Rβ and/or IL-2Rγc ligands, for example to enhance the therapeutic efficacy of compounds comprising IL-2Rβγc agonists or IL-2Rβ and/or IL-2Rγc ligands, or The gastrointestinal tract of compounds comprising IL-2Rβγc agonists or IL-2Rβ and/or IL-2Rβ and/or IL-2Rγc ligands by co-administering any of the foregoing pharmaceutical compositions with one or more active agents. may increase absorption or diffusion from cells into the systemic circulation, or may inhibit degradation of compounds, including IL-2Rβγc agonists or IL-2Rβ and/or IL-2Rγc ligands, in the subject's blood. Pharmaceutical compositions comprising IL-2Rβγc agonists or compounds comprising IL-2Rβ and/or IL-2Rγc ligands enhance the therapeutic efficacy of IL-2Rβγc agonists or compounds comprising IL-2Rβ and/or IL-2Rγc ligands It can be co-administered with an active agent that has a pharmacological effect.

IL-2Rβγc agonists, compounds comprising IL-2Rβ and/or IL-2Rγc ligands, or pharmaceutical compositions comprising any of the foregoing are known for treating an inflammatory or autoimmune disease in a patient; Or it may be administered in conjunction with agents that are believed to be effective.

An IL-2Rβγc agonist, a compound comprising IL-2Rβ and/or an IL-2Rγc ligand, or a pharmaceutical composition comprising any of the foregoing, is an agent known or believed to interfere with proliferation. They can be administered together. IL-2Rβγc agonists, compounds comprising IL-2Rβ and/or IL-2Rγc ligands, or pharmaceutical compositions comprising any of the foregoing are known to or believed to interfere with mitosis. It can be administered in conjunction with a drug. An IL-2Rβγc agonist, a compound comprising IL-2Rβ and/or an IL-2Rγc ligand, or a pharmaceutical composition comprising any of the foregoing agents known or believed to interfere with DNA replication can be administered in conjunction with IL-2Rβγc agonists, compounds comprising IL-2Rβ and/or IL-2Rβ and/or IL-2Rγc ligands, or pharmaceutical compositions comprising IL-2Rβγc agonists are known to interfere with or interfere with DNA repair. It may be administered in conjunction with an agent that is believed to be

The compounds provided by this disclosure are useful for understanding the biological role of IL-2, including evaluating the many factors that influence and are thought to be affected by IL-2 production and receptor binding processes. can be useful in vitro as a tool for The compounds are also useful in developing other compounds that bind to and activate IL-2R. This is because the present compounds provide useful information regarding structure-activity relationships that should facilitate such development.

The compounds are also useful as competitive binders in assays for screening new IL-2 receptor antagonists. In such assay embodiments, the compounds of the invention can be used without modification, or can be modified in various ways. For example, it can be by labeling, such as covalently or non-covalently binding moieties that directly or indirectly provide a detectable signal. In any of these assays the materials can be labeled either directly or indirectly. Direct labeling possibilities include, for example, radiolabels such as ¹²⁵ I, enzymes such as peroxidase and alkaline phosphatase, and labeling groups such as fluorescent labels that allow monitoring of fluorescence intensity, wavelength shifts, or changes in fluorescence polarization. mentioned. Possibilities for indirect labeling include biotinylation of one component followed by binding to avidin bound to one of the above labeling groups. A compound may also include a spacer or linker when the compound is attached to a solid support.

Based on their ability to bind to the IL-2 receptor, the peptides provided by this disclosure can be used as reagents for detecting the IL-2 receptor, e.g., in living cells, fixed cells, biological fluids. , in tissue homogenates, in purified and native biological materials. For example, by labeling such peptides, cells with IL-2 receptors on their surface can be identified. Additionally, based on their ability to bind IL-2 receptors, the peptides of the invention can be used for, eg, in situ staining, FACS (fluorescence activated cell sorting), Western blotting, and ELISA. In addition, based on their ability to bind the IL-2 receptor, the peptides provided by this disclosure may be used for receptor purification or to express IL-2 receptor on the cell surface (or in permeabilized cells). can be used to purify cells that

The compounds provided by this disclosure are also available as commercial reagents for various medical research and diagnostic applications. Such uses include, for example, (1) use as a calibration standard to quantify the activity of candidate IL-2 agonists in various functional assays, (2) expansion and growth of IL-2 dependent cell lines. (3) use in structural analysis of the IL-2 receptor by co-crystallization; (4) use to investigate the mechanism of IL-2 signaling/receptor activation; ) other research and diagnostic applications in which the IL-2 receptor is preferably activated or such activation is conveniently calibrated against a known amount of IL-2R agonist.

The following examples detail the methods used to determine the activity of peptides with IL-2Rβ and IL-2Rγc subunits. It will be apparent to those skilled in the art that many modifications, both to materials and methods, can be practiced without departing from the scope of this disclosure. In the Examples, IL-2Rβ subunit refers to human IL-2Rβ (CD122 protein, Fc tag) (27-239), Accession No. NP_000869.1, available from ACRObiosystems, Inc.; , under product number ILB-H5253. "IL-2Rγc subunit" refers to human IL-2Rγc (CD132 protein, Fc tag) (23-254), Accession No. AAH14972, available from ACROBiosystems, Inc.; , under product number ILG-H5256.

Example 1
Phage Display pIII Library Panning Procedure for Fc-Receptor Fusions on Magnetic Beads (Acid Elution) For each library sample, 50 μL of Protein G Dynabeads® (Invitrogen) were used. After resuspending the stock bottle, the desired volume of beads was transferred to a sterile microfuge tube and applied to the magnet.

Beads were removed on a magnet and beads were washed with 1 mL of PT buffer (1×PBS, 0.05% Tween®20).

The supernatant was removed and 1 mL of PBS + 1% BSA + 0.05% Tween® 20 was added and mixed for at least 1 hour at 25°C to block the beads.

The tube was applied to the magnet and the blocking solution was removed. For each library tested, 5 μg of Fc-fusion receptor of interest was added to each library sample for each round to bring the total volume to at least 400 μL. The samples were mixed for at least 1 hour at 25°C. The sample was applied to the magnet and the supernatant was removed.

200 μL of PT buffer was added for every 50 μL of beads. Samples were mixed thoroughly and 200 μL aliquots were transferred to pre-labeled tubes for each library to be screened. An additional 500 μL of PT was added to each tube, the samples were mixed and then applied to the magnet. A total of 700 μL/tube was used for washing.

1 mL aliquot of library removed from −20° C. freezer. 100 μL (1000 μL of 10×BT buffer (5% BSA, 0.5% Tween® 20 in 1×PBS)) was added to each tube and vortexed. Samples were then incubated for at least 2 hours on a rotator at 4° C. For additional screening rounds, 1 mL aliquots of amplification beads from the previous round from each library were used. was collected with a magnet, the phage solution was removed, the beads were washed twice with 1 mL of PT buffer, 500 μL of PT buffer was added, and the suspension was transferred to a clean tube.The beads were collected with a magnet. and removed the final wash.

475 μL of phage elution buffer was added to each well (0.2 M glycine-HCL, pH 2.2, 1 mg/mL BSA). Samples were incubated on a rotator for 10 minutes at 25°C. Beads were collected on a magnet and eluted phages were transferred to clean tubes.

25 μL of Neutralization Buffer (2M Tris base) was added to 475 μL of eluate. Neutralized samples were kept at 4° C. until ready for amplification of TG1 cells. Samples were stored at -20°C after screening. 50 μL (approximately 10% of total volume) was transferred to a 1.5 mL microcentrifuge tube and stored at −20° C. for use in deep sequencing.

Example 2
TG1 Culture and Library Amplification Fresh TG1 (or OmniMax) cultures were grown for approximately 1-1.5 hours after adding the library to the beads. 2X-YT medium (10 mL) was placed in a 50 mL Falcon® tube. 200 μL of TG1 was added to the Falcon tube overnight. 2X-YT medium (600 μL) was placed in an OD600 blank cuvette. Cultures were grown at 250 rpm and 37° C. and the first OD measurement was taken after 60 minutes. TG1 cells should be in log phase with an OD600 of 0.5-0.7 at the time of use.

Eluted phage (400-450 μL) was added to 1 mL of TG1 cells at an OD600 of 0.5-0.7 in a 50 mL Falcon® tube. Phages and TG1 cells were incubated for 30 min at 37° C. without shaking. Approximately 50-100 μL was deposited for titration and characterization.

2YT medium (10.5 mL) was added to 12 μL of carbenicillin (carb) (100 μg/mL to make 100 mg/mL) and 24 μL of 50% glucose (to make 0.1% glucose) and the cells was incubated at 37° C. with shaking at 250 rpm for 1 hour.

M13K07 helper phage (5×10 ¹⁰ pfu, 24 μL stock, 2×10 ¹² pfu/mL) was then added and vortexed to mix. Phages and cells were incubated for 30 minutes at 37°C without shaking.

Kanamycin was diluted to 3 mg/mL and arabinose to 2.4% in 2YT medium/carbenicillin-100/0.1% glucose and 100 μL was added to each amplification. The mixture was incubated overnight at 37°C and 250 rpm.

The culture was transferred to a 50 mL high speed VWR centrifuge tube and centrifuged at 8,000 g for 15 minutes at 4° C. in a JSP-F50C centrifuge to pellet the cells.

Transfer the supernatant to a 50 mL high speed VWR centrifuge tube, add 0.2 volumes of PEG/NaCl (volume multiplied by 0.25 mL to give 3 mL of PEG/NaCl for 12 mL amplification), mix, Incubated on ice for 30 minutes.

Cells were then centrifuged at 10,500 g at 4° C. for 15 minutes in a JSP-F50C centrifuge. The supernatant was removed and the phage pellet was resuspended in a total of 1 mL PBT (1×PBS, 0.05% Tween® 20, 0.5% BSA) by pipetting.

Samples were transferred to Eppendorf tubes, vortexed, and centrifuged at 12,000 rpm for 30 seconds. The supernatant was transferred to a clean Eppendorf tube and stored at 4°C. This amplified phage sample (250-500 μL) was used for the next round of screening.

Example 3
Preparation of Cultures from Individual Colonies A 96-well deep well plate was filled with 1 mL of 2YT broth/ampicillin-50/0.1% glucose. 96 colonies were plated into wells using a P20 tip. A tip was left in the well to mark the position. After the entire plate was completed, the tip was removed using a multichannel pipette. The plate was covered with a breathable film.

The inoculated plate(s) were incubated in a shaker at 37° C. until the culture became cloudy, typically within 4 hours at 250 rpm.

Remove the plate(s) from the incubator and transfer 50 μL of culture from each well to another deep well block designated as "archive block" containing 1 mL of 2YT broth/ampicillin-50/0.1% glucose. I took it out. Plate(s) were covered with breathable film and incubated overnight at 37° C. and 250 rpm.

After overnight incubation, M13K07 helper phage was added to 2×10 ¹⁰ pfu/mL in 2YT broth/ampicillin-50/0.1% glucose (making 6.0 mL per block). 50 μL of diluted M13K07 was added to each culture well in the deep well block. Deep well blocks were covered with breathable film and incubated at 37° C. and 250 rpm for 30 minutes.

Dilute kanamycin to 0.5 mg/ml and arabinose to 0.4% in 2YT broth/ampicillin-50/0.1% glucose (make 6.0 ml per block) and add 50 μL to each well. was added to Plates were covered with breathable film and incubated overnight at 37° C. and 250 rpm.

The 'archive block' culture was removed from the incubator and 50 μL transferred to a 96-well plate containing 50 μL of 50% glycerol. Plates were sealed in foil and stored at -80°C. The remaining culture in the block was covered with a foil seal and stored at 4°C.

Blocks were centrifuged and inoculated with M13K07 at 4000 rpm for 15 minutes. Avoiding the bacterial pellet, 850 μL of phage supernatant was transferred to a fresh deep well plate, covered with a foil seal and stored at 4°C.

Example 4
ELISA Protocol for Fc-Receptor Fusions For each block to be assayed, 1×96-well ELISA plates were coated with 50 μL/well of Fc-receptor fusions (1 μg/mL in PBS). Wells were incubated at 25° C. for at least 1 hour.

Fc-receptor fusions were removed from each well. 300 μL of blocking buffer (1×PBS, 1% BSA) was added to each well of the receptor-coated plate. The plate was covered with film and left at 37°C for 1 hour or at 4°C overnight.

Plates were washed four times with PT (1×PBS, 0.05% Tween® 20) buffer.

50 μL of PBT was added to each well. 50 μL of phage supernatant from the block was added to each well and incubated for 1 hour at 4°C.

Plates were washed four times with cold PT.

To each well was added 100 μL of anti-M13-HRP antibody diluted 1:5000 in cold PBT. Wells were incubated for 1 hour at 4°C.

Plates were then washed four times with cold PT.

50 μL of TMB was then added to each well and the wells were incubated at 25° C. for 1-10 minutes. 50 μL of 'stop' solution was added and the plate was read at 450 nm.

Example 5
Evaluation of peptide heterodimer ability to dimerize IL-2Rβγc and activate IL-2 responsive cells Identification of ligands exhibiting IL-2Rβ and IL-2Rγc binding activity, followed by IL-2R agonist activity Compounds were identified. This involved assessing the ability of peptides to dimerize and signal IL-2Rβγc subunits in cell-based assays. Dimerization is a necessary but not sufficient step in the activation of receptor signaling. To assess agonist activity in cell-based assays, IL-2 responsive cell lines were tested for phosphorylation of STAT5, an indicator of IL-2 signaling. Compounds that showed IL-2Rβγc agonist activity in these cell lines were then treated with IL-2R agonism and cells expressing IL-2Rβγc subunit but low or no IL-2Rα (CD25) subunit expression. The desired selectivity in favor of type activation was tested in primary human peripheral blood mononuclear cells (PBMC).

Dimerization potential was assessed using the β-Gal complementation system. In the β-Gal complementation system, a portion of the intracellular domain of each respective IL-2 receptor subunit is replaced with a functionally complementary fragment of β-Gal and regains catalytic activity when in sufficient proximity. Cells expressing these constructs produce β-Gal activity with an IC ₅₀ of approximately 26 nM when treated with IL-2 (see DiscoverX product specifications). All synthetic, potentially agonistic peptides were tested using this assay.

Candidate compounds were scored for induction of STAT5 phosphorylation in two cell lines. (1) NK-92 cells, a human cell line that expressed all three IL-2 receptor subunits and was responsive to the IL-2Rβγc biased variant as well as wild-type IL-2; and (2) TF- 1β cells, derived from the human erythroleukemia line TF-1, naturally expressing only IL-2Rγc, were engineered to be IL-2 responsive by transfection of IL-2Rβ. TF-1β was constructed and IL-2R subunit expression levels in both cell lines were verified by QPCR and FACS analysis.

Compounds were tested in both cell lines. A dose-response assay was performed to determine the _IC50 of test compounds and compare test compounds to IL-2 as an indicator of IL-2Rβγc receptor bias.

Conferring IL-2 responsiveness to TF-1 cells by transfection and expression of the IL-2Rβ subunit demonstrates that peptide agonist activity is dependent on the presence of the IL-2Rβ subunit. Neutralizing antibodies against IL-2 and IL-15 (R&D Systems) was included in the activity assay and shown to inhibit IL-2 and IL-15 activity, but not to affect the agonist activity of the test peptides.

Compounds demonstrating IL-2R agonist activity on cell lines are tested on human primary immune cells, PBMC collected from individual donors (commercially available from Lonza) and optionally on purified CD4+ cells (Lonza). bottom. A substantial portion of PBMC from normal donors responded to IL-2. To assess IL-2 agonist activity of test compounds, cells were exposed to compound or IL-2 and scored for STAT5 phosphorylation by Western blot analysis. Those compounds demonstrating STAT5 activation of PBMCs were subjected to a follow-on assay designed to assess the subunit bias of the compounds compared to IL-2. This assay was scored by a FACS-based protocol that allowed detection of both intracellular pSTAT5 as an indicator of IL-2R activation and the IL-2Rα subunit, cell-surface CD25, over a 30-minute study. It involved determining the dose response of compound and IL-2 (1-1000 IU). Cells expressing the three IL-2R subunits, IL-2Rαβγc, bind IL-2 with very high affinity (approximately 10 pM) and are therefore sensitive to low concentrations of IL-2, whereas IL-2R Cells expressing only 2Rβγc (approximately 1 nM affinity) require exposure to very high IL-2 levels for activation. Compounds provided by the present disclosure were selected to bind to IL-2Rβ and IL-2Rγc subunits, but not to IL-2Rα subunits, so the potency of the compounds was limited to IL-2Rα on cells. and a comparison of the response profiles of cells treated with compounds provided by this disclosure or treated with IL-2 should reveal any bias. is.

Example 6
Identified Peptides Four probabilistic libraries, each containing approximately 10 ¹⁰ independent recombination, with each clone potentially representing a unique peptide sequence, were isolated from individual human IL-2Rβ and IL-2Rγc subgroups. Screened for binding to the unit. In screening these four initial libraries against the Rβ subunit extracellular domain (ECD), 98 unique peptide clones were identified and confirmed as ligands of IL-2Rβ. These IL-2Rβ ligands can be grouped into at least two sequence families, family 1 and family 2.

Fifteen unique peptide sequences were identified in screening the four initial libraries for the γc subunit, which in the first analysis of this limited number of clones exceeded one Suggesting that they may represent distinct sequence families. At the current level of resolution, these peptide sequences appear to have no sequence similarity to human IL-2. Most of the peptides recovered to date have been tested for binding to the IL-2Rβ and/or IL-2Rγc subunits, and all peptides tested show that they have a weak affinity (up to 10-50 μM was found to bind only to selected subunits (by phage ELISA, which can detect .

Analysis of samples of the identified peptides was performed to determine human and mouse receptor specificity. Although initially selected on the human IL-2 receptor, neither IL-2Rβ nor IL-2Rγc ligands bound exclusively to the corresponding human subunits.

Peptides having SEQ ID NO:377-SEQ ID NO:399 have been synthesized and evaluated for IL-2Rβ and IL-2Rγc activity. Peptides having SEQ ID NOS:377-385 include IL-2Rβ ligands and peptides having SEQ ID NOS:386-399 include IL-2Rγc ligands.

Example 7
Preparation of NK-92 Cells for STAT5 Activation Test NK-92 cells were seeded in 24-well plates at 4×10 ⁵ cells in 1 mL starvation medium (SM) and incubated overnight at 37° C., 5% CO _{2 .} bottom. Starvation medium contained RPMI 1640 + 20% FBS + 2 mM L-glutamine + 1 mM NaPyr + 10 mM HEPES + 0.1 mM BME (no rhIL-2 supplement).

Treatment mixtures were prepared with 1 μg/mL anti-hIL-2 neutralizing antibody (0.2 mg/mL stock solution) or goat IgG control (1 mg/mL stock solution).

The treatment mixture and antibody mixture were added to the cells for 30 minutes at 37°C, 5% _CO2 . Each sample was then transferred to a 1.5 mL microcentrifuge tube and spun down at 1,500 RPM for 5 minutes. Cells were washed in 1 mL PBS and centrifuged again.

A phosphatase and protease inhibitor cocktail (Thermo #78442) was added to the mPER buffer at a 1:100 dilution. After pelleting the cells, 50 μL of mPER buffer was added to each sample and homogenized by repeated pipetting.

Lysates were centrifuged at 14,000 RPM for 5 minutes at room temperature. The supernatant was transferred to a clean tube and stored frozen at -80°C.

Human IL-2 antibody (goat IgG) was obtained from R&D Systems at No. AF-202-NA and a normal goat IgG control from R&D Systems at No. Obtained at AB-108-C, anti-STAT5 antibody (rabbit), Cell Signaling No. 94205S, anti-pSTAT5 antibody (rabbit), Cell Signaling No. 4322S, and goat anti-rabbit IgG-HRP from Jackson Immunoresearch, no. Obtained at 111-035-144.

Antibodies, treatments, and working stock preparations for each of the samples are provided in Table 1. Compounds A and B are IL-2Rβγc agonists provided by this disclosure.

Samples were applied to Western blots and the results are shown in FIG. In the Western blot shown in FIG. 1, samples #1-10 were probed with anti-pSTAT5 antibody, samples #1-10 were probed with anti-pSTAT5 antibody, samples #11-20 were probed with anti-pSTAT5 antibody, sample #11 ~20 were probed with an anti-STAT5 antibody. Treatment reagents included anti-STAT5 antibody (rabbit), Cell Signaling, No. 94205S, anti-pSTAT5 antibody (rabbit), Cell Signaling, no. 4322S, and goat anti-rabbit IgG-HRP, Jackson Immunoresearch, no. 111-035-144 was included.

Compounds A and B induced STAT5 phosphorylation (pSTAT5) (in a dose-responsive manner). As shown in Figure 1, the first two sets of three lanes contain two test compounds and the next three lanes are the IL-2 positive control. The final lane (10) is "starved" cells showing no pSTAT5 background in the starting cell population. Compound A refers to the heterodimer of SEQ ID NO:58 and SEQ ID NO:224. Compound B refers to the heterodimer of SEQ ID NO:58 and SEQ ID NO:237.

At the top of the lane are the results of a control experiment designed to rule out the possibility that the test samples were contaminated with IL-2. To assess contamination, the same procedure was performed as for the lanes described above, except that an IL-2 neutralizing antibody was added to all samples. As shown in lanes 7-9, the IL-2 control was inhibited by this treatment, while the test compound lanes (1-6) gave similar results to the minus Ab lanes, indicating that the activity in the compound lanes was higher than that of IL-2. We have demonstrated that it is not due to contamination. In addition, the lane 9 control has a high amount of DMSO (1%), a diluent for the peptide compound. Addition of DMSO does not cause STAT5 phosphorylation.

Activation of STAT5, ERK1/2 and AKT in NK-92 cells by IL-2Rβγc agonists A and B provided by this disclosure is shown in FIG. IL-2Rβγc agonist A refers to the heterodimer of SEQ ID NO:58 and SEQ ID NO:224. IL-2Rβγc agonist B refers to the heterodimer of SEQ ID NO:58 and SEQ ID NO:237.

The pSTAT5 dose-response curves of L-2Rβγc agonists A and B in NK-92 cells are shown in FIGS. 4A and 4B.

Results of NK-92 cell proliferation assays using IL-2Rβγc agonists A and B, rhIL-2 and other peptides CG are shown in Figures 5A-5C. Compounds AG are defined as in Table 2.

To perform the assay, NK-92 cells were seeded in starvation medium at 20,000 cells/well in 96-well plates. Treatments were added to each well in 3-fold serial dilutions with peptides having a maximum concentration of 10 μM and rhL-2 having a maximum concentration of 6.67 nM. Cells were then incubated at 37° C. for 48 hours. CellTiter-Glo® reagent was added and the cells were incubated at 25° C. for 10 minutes before taking a luminescence reading. FIG. 5A shows RLU for peptide samples, FIG. 5B shows rhIL-2 samples, and FIG. 5C shows an overlay of FIGS. 5A and 5B.

Example 8
Preparation of TF-1β and TF-1 cells for STAT5 activation studies TF-1β and TF-1 parental cells were counted. Cells were harvested and 2.5×10 ⁶ cells were pelleted at 200×g for 5 minutes. Pelleted cells were washed with 25 mL RPMI without additives.

TF-1β and TF-1 parental cells were seeded at 5×10 ⁵ cells in 5 mL starvation medium (SM) in T25 flasks and incubated overnight at 37° C. in 5% CO ₂ with flasks upright.

TF-1β and TF-1 parental cells were counted and viability determined. If necessary, cells were diluted to 5×10 ⁵ cells/mL in SM, then 1 mL of suspension was added at 6 wells/cell line in 24-well dishes and incubated at 37° C. with 5% CO ₂ .

Treatments (see Example 7) were added to cells for 30 min at 37° C. under 5% CO ₂ . Treated cells were transferred to a 1.5 mL microcentrifuge tube and spun down at 1,500 RPM for 5 minutes. Cells were washed in 1 mL of PBS, centrifuged again, and the supernatant was aspirated. Treatment reagents included anti-STAT5 antibody (rabbit), Cell Signaling, No. 94205S, anti-pSTAT5 antibody (rabbit), Cell Signaling, no. 4322S, and goat anti-rabbit IgG-HRP, Jackson Immunoresearch, no. 111-035-144 was included.

A phosphatase and protease inhibitor cocktail (Thermo No. 78442) was added to the mPER buffer at a 1:100 dilution. After pelleting the cells, 50 μL of mPER buffer was added to each sample and the mixture was homogenized by repeated pipetting.

The lysate was centrifuged at 14,000 RPM for 5 minutes at 25°C. The supernatant was transferred to a clean tube and stored frozen at -80°C.

Antibodies, treatments, and working stock preparations for each of the samples are provided in Table 3. Compounds A and B are IL-2Rβγc agonists provided by this disclosure.

Samples were applied to Western blots and the results are shown in Figure 2A.

RT-qPCR gene expression profiling comparing several TF-1β cell populations (with varying concentrations of G418) transfected with full-length IL-2Rβ used for peptide simulation pSTAT5 evaluation are shown in Table 4 and Figure 2B. . In FIG. 2B, relative expression levels were normalized to GAPDH=1×10 ⁶ copies (REL=1×10 ⁶ ×2 ^{(Ctgraph−Cttarget)} ).

Example 9
pH-Selective Screening IL-2Rβ subunits were screened with two peptide libraries to identify peptides exhibiting pH-dependent affinity for the receptor subunits. The screening approach utilized cycles of binding and elution under various acidic and neutral pH conditions.

Example 10
Phage Screening ELISA Protocol for pH Dependence A phage ELISA at two target pH values was used to determine phage binding to IL-2Rβ-GPI and percent change in binding at pH 7.4 versus pH 6.0. was calculated. Results for several peptides are shown in FIG. Well G03 is an example of a phage that exhibits pH-dependent binding to IL-2Rβ.

Figure 7 shows pH dependent binding (17E01, 17G03, 18A08, 17H07) compared to non-pH dependent clones (negative clones) that showed similar binding affinities at both pH 6.0 and pH 7.4. Examples of phage clones are shown.

Example 11
ELISA protocol for pH-dependent phage titration The ELISA screening protocol described in Example 10 was used with the following differences: (1) All 96-well ELISA plates contained IL-2Rβ-GPI targets; , (2) titration of phage supernatants was prepared in two different PBT pH buffers (pH 6.0 and pH 7.4).

Phage titration was performed in 96-well polypropylene plates using the following procedure. Three-fold dilutions of phage in PBT pH 6 and pH 7.4 buffers were prepared. 100 μL of diluted phage was transferred to target-coated assay plates and incubated at 4° C. for 1 hour.

The pH 6.0 wells were washed three times with cold PT pH 6.0 and the pH 7.4 wells were washed twice with cold PT pH 7.4.

Bound phage were detected with anti-M13-HRP.

Binding curves for pH-independent peptides that showed similar binding at pH 6.0 and pH 7.4 are shown in Figure 8A, and binding curves for pH-dependent clones are shown in Figure 8B.

Example 12
ELISA protocol for biotinylated peptide pH dependent binding (IL-2Rβ/Fc-receptor binding/multivalent):
For each peptide assayed, 16 ELISA plate wells were coated with neutravidin (10 μg/mL in PBS pH 7.2) at 50 μL/well. Coated wells were incubated at 25° C. for at least 1 hour.

Neutravidin was removed from each well. 300 μL of blocking buffer (1×PBS pH 7.2, 1% BSA) was added to each well of the neutravidin-coated plate. All plates were covered and kept at 25°C for 1 hour or 4°C overnight.

Incubated plates were washed four times with PT (1×PBS pH 7.2, 0.05% Tween® 20) buffer.

Biotinylated peptides were diluted to 1 μM in PBT pH 7.2 buffer and 50 uL added to appropriate 16 wells (8 for each binding pH). Plates were incubated at 25° C. for at least 1 hour.

Duplicate titrations of IL-2Rβ-Fc protein were prepared in polypropylene plates using PBT pH 6.0 and pH 7.4, starting at 2 μg/mL and diluting 3-fold.

Plates were washed four times with PT (1×PBS pH 7.2, 0.05% Tween® 20) buffer.

50 μL of IL-2Rβ-Fc protein dilution was added to assay plates buffered at pH 6.0 or pH 7.4 and incubated at 4° C. for 1 hour.

Incubated plates were added to the corresponding pH buffer PT (50 mM PBS pH 6.0, 0.05% Tween® 20 or 50 mM PBS (pH 7.4, 0.05% Tween® 20). ) three times.

Goat anti-huIgG-HRP diluted 1:2500 in 50 μL of cold PBT (pH 6.0) was added to each well. Plates were then incubated for 1 hour at 4°C.

Plates were then washed four times with cold PT pH 6.0.
50 μL of TMB was then added to each well and the wells were incubated at 25° C. for 1-10 minutes. 50 μL of 'stop' solution was added to each well and the plate was read at 450 nm.

IL-2Rβ-Fc binding curves comparing peptides exhibiting pH-independent binding at pH 6.0 and pH 7.4 with pH-dependent peptides are shown in Figures 9A and 9B, respectively. Fc binding is a multivalent interaction and thus an overestimation of monovalent binding.

Example 13
Specific Heterodimeric IL-2Rβγc Agonists The C-terminus of IL-2Rγc ligand having SEQ ID NO: 58 is combined with the C-terminus of IL-2Rβ ligand having SEQ ID NO: 224 and four amino acids using standard click chemistry methods. The heterodimer was constructed by linking with a linker with a length of about 34 Å using .

Heterodimers were incubated with NK92 cells and STAT5 phosphorylation was measured as a function of concentration using methods described in Examples 7 and 8. The results are shown in FIG.

Example 14
Specific Heterodimers An IL-2Rβ ligand provided by the present disclosure was combined with a linker containing 3-6 amino acids and an IL-2Rγc ligand provided by the present disclosure using standard click chemistry methods. A heterodimer was constructed by ligation.

Example 15
pSTAT5 activation in NK92 and TF1-β cells Expression levels of IL-2R subunits in NK92 and TF-1β cells were determined using RT-qPCR gene expression profiling. Results are presented in Table 5.

The heterodimers of Example 14 were incubated with NK92 cells and TF-1β cells and STAT5 phosphorylation was measured as a function of concentration using methods described in Examples 7 and 8. Results are presented in Table 5.

Example 15
NK-92 Cell Proliferation NK-92 cell proliferation was determined using the procedure described in Example 7.

The EC ₅₀ for NK-92 cell proliferation for the heterodimer of Example 14 was less than 10 ⁻⁸ M.

Example 16
pSTAT5 Activation in Human T Cells pSTAT5 activation of resting CD8 T cells, resting CD4 T cells, and resting Treg (CD25hi CD127lo) cells by the heterodimer of Example 14 was was determined using

Incubation time was 30 minutes and pSTAT5 was measured by flow cytometry.

The heterodimer of Example 14 showed similar potency in different human T cell populations tested. In comparison, IL-2 has a potency bias towards Tregs and CD4 T cells.

Example 17
STAT5 phosphorylated CD8 T cells in resting and activated human CD-8 T cells were analyzed by STEMCELL™ Technologies Inc. Whole PBMC pools were isolated using the EasySep™ Human CD8+ T cell isolation kit commercially available from .

Treg cells were obtained from STEMCELL™ Technologies Inc. Whole PBMC pools were isolated by the EasySep™ Human CD4+CD127lowCD25+ Regulatory T Cell Isolation Kit, commercially available from .

For resting groups, pSTAT5 assays were pre-performed and lysates were frozen at -180°C until ELISA measurements.

For the activation group, cells were resuspended at 10 ⁶ cells/mL in CTS OpTmizer™ medium and prepared for CD28/CD3 activation.

For CD8 cells, a solution of 1 μg/μL aCD28 antibody was added to the cell suspension and seeded at 2-3 mL/well in 6-well plates pre-coated with 1 μg/mL aCD3 antibody.

For Treg cells, a solution of 10 μg/mL CD28 antibody was added to the cell suspension and seeded at 2-3 mL/well in 6-well plates pre-coated with 10 μg/mL CD3 antibody.

Cells were incubated for 3 days.

After incubation, cells were resuspended at 5×10 ³ cells/mL in fresh CTS™ OpTmizer™ media (ThermoFisher Scientific) and seeded at 2-3 mL/well in 6-well plates. Reseeded cells were incubated for 2 days.

Control and test compounds were added to the cells at various concentrations and incubated for 30 minutes at 37°C.

Cells were harvested, counted, lysed and assayed for pSTAT5 activation.

For resting CD8 T cells, the _EC50 for STAT5 phosphorylation was less than ^10-8M .

For activated CD8 T cells, the _EC50 for STAT5 phosphorylation was less than ^10-8M .

Example 18
STAT5 Phosphorylation in Resting and Activated Human Treg Cells Using the same procedure described in Example 17, STAT5 phosphorylation was measured.

For resting Treg cells, the _EC50 for STAT5 phosphorylation was less than ^10-8M .

For activated Treg cells, the _EC50 for STAT5 phosphorylation was less than ^10-8M .

Example 19
Binding of IL-2Rα and IL-2Rβ IL-2R subunit expression levels in TF-1 parental and TF-1β cells were determined using RT-qPCR gene expression profiling. Results are presented in Table 6.

TF-1 parental and TF-1β cells were incubated with the heterodimer of Example 14 and STAT5 activation was measured as a function of concentration. Results are presented in Table 6.

STAT5 was not activated for TF-1 parental cells.

For activated Treg cells, the _EC50 for STAT5 phosphorylation was less than ^10-8M .

These results demonstrate that activity of the heterodimer of Example 14 requires the presence of IL-2Rβ subunits. Also, in a separate experiment, it was determined that the activity of the heterodimer of Example 14 was not blocked by IL-2 and IL-15 antibodies.

Example 20
Competitive Binding Competitive binding assays were performed to characterize the IL-2R binding sites of specific IL-2Rβ and IL-2Rγc ligands.

Representative phage clones displaying peptides from the IL-2Rβ ligand family were allowed to bind to the extracellular domain (ECD) of IL-2Rβ immobilized on microtiter wells. Phage binding was performed in the presence and absence of synthetic test peptides to determine whether the phage peptide and test peptide competed for binding to the same site on IL-2Rβ. Synthetic test peptides were selected to represent peptides from the IL-2Rβ ligand family, as well as positive and negative control peptides. IL-2Rβ ligand family sequences and specific IL-2Rβ ligands evaluated are provided in Table 7.

IL-2Rβ ligands had binding affinities (IC ₅₀ ) for IL-2Rβ subunits less than 10 μM and binding affinities (IC ₅₀ ) for IL-2Rγc subunits greater than 100 μM.

Phage binding to immobilized IL-2Rβ ECD was scored by staining with antibodies to phage coat proteins (anti-phage Abs), labeled secondary antibodies to anti-phage Abs, and reading the OD in a microtiter plate optical reader. It was detected by

The ELISA signal for each phage binding in the presence and absence of test peptides was compared to determine which synthetic peptides competed for binding to IL-2Rβ subunits. Those peptide pairs that exhibited competitive binding (ie, cross-inhibition) were thought to bind at the same functional site on the IL-2 receptor. Results are presented in Table 8.

IL-2Rβ ligands did not competitively bind to the IL-2 and IL-2Rβ subunit binding sites.

Similar studies were performed to assess binding of IL-2Rγc ligands. IL-2Rγc ligand family sequences and specific IL-2Rγc ligands evaluated are provided in Table 9.

IL-2Rγc ligands had binding affinities (IC ₅₀ ) for IL-2Rγc subunits less than 10 μM and binding affinities (IC ₅₀ ) for IL-2Rβ subunits greater than 100 μM.

Results are presented in Table 10.

Aspects of the Invention The invention is further defined by the following aspects.

Aspect 1. An IL-2Rβ ligand, wherein the IL-2Rβ ligand exhibits a binding affinity for human IL-2Rβ subunits of less than 100 μM.

Aspect 2. An IL-2Rβ ligand according to aspect 1, wherein the IL-2Rβ ligand comprises 5-30 amino acids.

Aspect 3. An IL-2Rβ ligand according to any one of aspects 1-2, wherein the IL-2Rβ ligand exhibits a binding affinity for human IL-2Rβ subunits of between 1 pM and 100 μM.

Aspect 4. An IL-2Rβ ligand according to any one of aspects 1-2, wherein the IL-2Rβ ligand exhibits a binding affinity for human IL-2Rβ subunits of between 0.1 μM and 50 μM.

Aspect 5. An IL-2Rβ ligand according to any one of aspects 1-2, wherein the IL-2Rβ ligand exhibits a binding affinity for human IL-2Rβ subunits of less than 100 μM.

Aspect 6. An IL-2Rβ ligand according to any one of aspects 1-5, wherein the IL-2Rβ ligand exhibits a binding affinity for mammalian IL-2Rβ subunits of less than 100 μM.

Aspect 7. An IL-2Rβ ligand according to any one of aspects 1-6, wherein the IL-2Rβ ligand exhibits a binding affinity of less than 100 μM for each of the human IL-2Rβ subunit and the human IL-2Rγc subunit.

Aspect 8. An IL-2Rβ ligand according to any one of aspects 1-7, wherein the IL-2Rβ ligand exhibits a binding affinity for the human IL-2Rα (CD25) subunit of greater than 100 μM.

Aspect 9. Any one of aspects 1-8, wherein the IL-2Rβ ligand exhibits a binding affinity for the human IL-2Rβ subunit that is at least 10-fold greater than the binding affinity of the IL-2Rβ ligand for the human IL-2Rα subunit. IL-2Rβ ligand as described in .

Aspect 10. the IL-2Rβ ligand comprises the amino acid sequence of formula (1) (SEQ ID NO:1), the amino acid sequence of formula (1a) (SEQ ID NO:2), or the amino acid sequence of formula (1b) (SEQ ID NO:3);
^{-X3 -X4 -X5 -X6 -X7 -X8 -X9 -X10} - (1)
^-X2 -CX3 ^{-X4 -X5 -X6 -X7 -X8 -X9 -X10 -CX11} - ( ^1a )
^{-X1 -X2} -CX3 ^{-X4 -X5 -X6 -X7 -X8 -X9 -X10 -CX11 -X12 -} (1b)
wherein X ¹ is selected from A, D, E, F, G, I, K, L, M, N, P, Q, S, T, V, W, and ^Y ; , C, D, E, F, G, H, K, L, N, P, R, S, T, W, and Y, and X ³ are selected from A, D, E, F, G, H , M, N, Q, R, S, T, W, and Y, and X ⁴ is selected from A, D, E, F, G, I, K, L, M, N, Q, R, S , T, V, and Y, X ⁵ is selected from A, G, I, Q, S, T, V, and W, X ⁶ is selected from A, D, E, G, H, K , L, M, N, P, Q, R, S, T, and V; X ⁷ is selected from F, I, K, L, Q, and V; X ⁸ is selected from D, F , G, H, M, N, W, and Y, X ⁹ is selected from A, D, E, M, P, Q, S, T, V, and W, and X ¹⁰ is selected from D , F, I, L, M, S, T, V, and Y, and X ¹¹ is selected from D, E, F, H, I, L, M, Q, S, T, V, W, and 10. An IL-2Rβ ligand according to any one of aspects 1-9, wherein X ¹² is selected from F, I, L, M, N, S, V, W, and Y.

Aspect 11. 11. An IL-2Rβ ligand according to aspect 10, wherein X ¹ is selected from F, I, L, M, and V.

Aspect 12. IL-2Rβ according to any one of aspects 1-11, wherein ^X2 is selected from D, E, F, G, H, L, N, P, R, S, T, W, and Y ligand.

Aspect 13. 13. An IL-2Rβ ligand according to any one of aspects 1-12, wherein ^X5 is A.

Aspect 14. 14. An IL-2Rβ ligand according to any one of aspects 1-13, wherein ^X6 is selected from D, E and Q.

Aspect 15. 15. An IL-2Rβ ligand according to any one of aspects 1-14, wherein ^X7 is selected from F, I, L, and V.

Aspect 16. 16. An IL-2Rβ ligand according to any one of aspects 1-15, wherein ^X8 is G.

Aspect 17. 17. An IL-2Rβ ligand according to any one of aspects 1-16, wherein ^X9 is selected from D, E and Q.

Aspect 18. 18. An IL-2Rβ ligand according to any one of aspects 1-17, wherein ^X10 is selected from F, I, L, M, V, and Y.

Aspect 19. An IL-2Rβ ligand according to any one of aspects 1-18, wherein X ¹¹ is selected from D and E.

Aspect 20. 20. An IL-2Rβ ligand according to any one of aspects 1-19, wherein ^X12 is selected from F, I, L, M and V.

Aspect 21. X ¹ is selected from F, I, L, M, and V and X ² is selected from D, E, F, G, H, L, N, P, R, S, T, W, and Y and X ³ is selected from A, D, E, F, G, H, M, N, Q, R, S, T, W, and Y, and X ⁴ is selected from A, D, E, F, is selected from G, I, K, L, M, N, Q, R, S, T, V, and Y; X ⁵ is A; X ⁶ is selected from D, E, and Q; X ⁷ is selected from F, I, L, and V, X ⁸ is G, X ⁹ is selected from D, E, and Q, X ¹⁰ is F, I, L, M, 11. An IL-2Rβ ligand according to aspect 10, wherein X ¹¹ is selected from V, and Y, X 11 is selected from D and E, and X ¹² is selected from F, I, L, M, and V.

Aspect 22. the IL-2Rβ ligand comprises the amino acid sequence of formula (1) (SEQ ID NO:1), the amino acid sequence of formula (1a) (SEQ ID NO:2), or the amino acid sequence of formula (1b) (SEQ ID NO:3);
^{-X3 -X4 -X5 -X6 -X7 -X8 -X9 -X10} - (1)
^-X2 -CX3 ^{-X4 -X5 -X6 -X7 -X8 -X9 -X10 -CX11} - ( ^1a )
^{-X1 -X2} -CX3 ^{-X4 -X5 -X6 -X7 -X8 -X9 -X10 -CX11 -X12 -} (1b)
wherein X ¹ is selected from amino acids, X ² is selected from amino acids, X ³ is selected from amino acids, X ⁴ is selected from amino acids, and X ⁵ contains a small hydrophobic side chain is selected from amino acids, X ⁶ is selected from amino acids, X ⁷ is selected from amino acids containing large hydrophobic side chains, X ⁸ is selected from amino acids containing small hydrophobic side chains, X ⁹ is selected from selected from amino acids containing polar neutral or acidic side chains, X ¹⁰ is selected from amino acids containing large hydrophobic side chains, X ¹¹ is selected from amino acids, and X ¹² contains large hydrophobic side chains An IL-2Rβ ligand according to any one of aspects 1-9, selected from amino acids.

Aspect 23. X ¹ is selected from amino acids containing large hydrophobic side chains, X ² is selected from amino acids, X ³ is selected from amino acids, X ⁴ is selected from amino acids, X ⁵ is selected from small hydrophobic is selected from amino acids containing side chains, X ⁶ is selected from amino acids containing polar neutral or acidic side chains, X ⁷ is selected from amino acids containing large hydrophobic side chains, and X ⁸ is selected from amino acids containing small hydrophobic side chains; selected from amino acids containing side chains, X ⁹ is selected from amino acids containing polar neutral or acidic side chains, X ¹⁰ is selected from amino acids containing large hydrophobic side chains, X ¹¹ is polar neutral 23. An IL-2Rβ ligand according to aspect 22, wherein X ^{12 is selected from amino acids containing an acidic side chain and X 12} is selected from amino acids containing a large hydrophobic side chain.

Aspect 24. X ¹ is selected from I, L, M, V, F, W, and Y, X ² is selected from amino acids, X ³ is selected from amino acids, X ⁴ is selected from amino acids, X ⁵ is selected from A, G, P, S, and T; X ⁶ is selected from H, N, Q, S, T, Y, D, and E; X ⁷ is selected from I, L, M , V, F, W, and Y, X ⁸ is selected from A, G, P, S, and T, and X ⁹ is H, N, Q, S, T, Y, D, and E, X ¹⁰ is selected from I, L, M, V, F, W, and Y, and X ¹¹ is selected from H, N, Q, S, T, Y, D, and E , X ¹² is selected from I, L, M, V, F, W, and Y.

Aspect 25. X ¹ is selected from I, L, M, V, F, W, and Y, X ² is selected from amino acids, X ³ is selected from amino acids, X ⁴ is selected from amino acids, X ⁵ is A; X ⁶ is selected from H, N, Q, S, T, Y, D, and E; X ⁷ is selected from I, L, M, V, F, W, and Y is selected, X ⁸ is G, X ⁹ is selected from H, N, Q, S, T, Y, D, and E, and X ¹⁰ is I, L, M, V, F, W , and Y, X ¹¹ is selected from H, N, Q, S, T, Y, D, and E, and X ¹² is selected from I, L, M, V, F, W, and Y 23. An IL-2Rβ ligand according to aspect 22, which is selected.

Aspect 26. 26. An IL-2Rβ ligand according to aspect 25, wherein X ¹ is selected from I, L, M, and V.

Aspect 27. 27. An IL-2Rβ ligand according to any one of aspects 25-26, wherein ^X2 is selected from D and E.

Aspect 28. 28. An IL-2Rβ ligand according to any one of aspects 25-27, wherein ^X6 is selected from Q, E and D.

Aspect 29. 29. An IL-2Rβ ligand according to any one of aspects 25-28, wherein ^X7 is selected from V, L and I.

Aspect 30. 30. An IL-2Rβ ligand according to any one of aspects 25-29, wherein ^X9 is selected from E, D and Q.

Aspect 31. An IL-2Rβ ligand according to any one of aspects 25-30, wherein X ¹⁰ is selected from L, V, I, and Y.

Aspect 32. 32. An IL-2Rβ ligand according to any one of aspects 25-31, wherein X ¹¹ is selected from D and E.

Aspect 33. 33. An IL-2Rβ ligand according to any one of aspects 25-32, wherein ^X12 is selected from L, I, and F.

Aspect 34. X ¹ is selected from L, I, F, and V ^; X ² is selected from D and E; X ⁶ is selected from Q, E, and D; is selected from I, ^{X 9} is selected from E, D, and Q; X ¹⁰ is selected from L, V, I, and Y; X ¹¹ is selected from D and E; 26. An IL-2Rβ ligand according to aspect 25, selected from L, I, and F.

Aspect 35. X ¹ is selected from F, I, M, and Y; X ² is selected from E, D, and R; X ³ is selected from and amino acids; X ⁴ is selected from amino acids; ⁵ is A, X ⁶ is selected from A, P, and Q, X ⁷ is selected from I and V, X ⁸ is G, and X ⁹ is selected from E and Q , X ¹⁰ is selected from I, L, and V; X ¹¹ is selected from E, D, and Q; and X ¹² is selected from I and L. .

Aspect 36. 10. An IL-2Rβ ligand according to any one of aspects 1-9, wherein the IL-2Rβ ligand comprises an amino acid sequence selected from any one of SEQ ID NOs:4-163.

Aspect 37. the IL-2Rβ ligand comprises an amino acid sequence selected from any one of SEQ ID NOs: 4-163, wherein the amino acid sequence is on the N-terminus, the C-terminus, or both the N-terminus and the C-terminus 37. An IL-2Rβ ligand according to aspect 36, which ends with the amino acids -GG.

Aspect 38. The IL-2Rβ ligand comprises an amino acid sequence selected from any one of SEQ ID NOs: 4-163, each amino acid independently having the following conservative substitutions: alanine (A), glycine ( G), amino acids with small hydrophobic side chains including proline (P), serine (S), and threonine (T); amino acids with hydroxyl-containing side chains including serine (S) or threonine (T); aspartic acid (D) and amino acids with acidic side chains, including glutamic acid (E); polar, including histidine (H), asparagine (N), glutamine (Q), serine (S), threonine (T), and tyrosine (Y). amino acids with neutral side chains; amino acids with basic side chains including arginine (R), lysine (K), or histidine (H); and isoleucine (I), leucine (L), methionine (M), valine 38. According to any one of aspects 36-37, comprising one or more of amino acids with large hydrophobic side chains, including (V), phenylphenyl (F), tyrosine (Y), or tyrosine (W). IL-2Rβ ligand of.

Aspect 39. the IL-2Rβ ligand comprises the amino acid sequence of formula (2) (SEQ ID NO: 164), the amino acid sequence of formula (2a) (SEQ ID NO: 165), or the amino acid sequence of formula (2b) (SEQ ID NO: 166);
^{-X15 -X16 -X17 -X18 -X19 -X20 -X21 -X22} - (2)
^{-X14 -CX15 -X16 -X17 -X18 -X19 -X20 -X21 -X22 -CX23} - (2a)
^{-X13 -X14 -CX15 -X16 -X17 -X18 -X19 -X20 -X21 -X22 -CX23 -X24} - (2b)
wherein X ¹³ is selected from A, D, E, G, N, Q, R, and V; ^{X 14} is selected from E, F, I, L, M, and Q; , D, G, L, and N, X ¹⁶ is selected from L, P, V, and Y, X ¹⁷ is selected from F, G, and M, and X ¹⁸ is selected from A, D , N, and Q; ^{X 19} is selected from F, I, L, S, V, W, and Y; X ²⁰ is selected from D and W; X ²² is selected from A, D, Q, and S; X ²³ is selected from I, L, Q, W, and Y; X ²⁴ is selected from E, F, I, L, T , V, and W. An IL-2Rβ ligand according to any one of aspects 1-9.

Aspect 40. 40. An IL-2Rβ ligand according to aspect 39, wherein ^X16 is V.

Aspect 41. An IL-2Rβ ligand according to any one of aspects 39-40, wherein ^X17 is G.

Aspect 42. 42. An IL-2Rβ ligand according to any one of aspects 39-41, wherein ^X20 is W.

Aspect 43. 43. An IL-2Rβ ligand according to any one of aspects 39-42, wherein ^X21 is P.

Aspect 44. X ¹³ is selected from E, N, and Q, X ¹⁴ is selected from I and M, X ¹⁵ is selected from D, L, and N, X ¹⁶ is V, and X ¹⁷ is , G, X ¹⁸ is selected from D and Q, X ¹⁹ is selected from V, W, and Y, X ²⁰ is W, X ²¹ is P, X ²² is 40. An IL-2Rβ ligand according to aspect 39, wherein ^X23 is selected from D and S, X23 is selected from L and Q, and ^X24 is selected from I, L, and V.

Aspect 45. An IL-2Rβ ligand according to any one of aspects 1-9, wherein the IL-2Rβ ligand comprises an amino acid sequence selected from any one of SEQ ID NOs:167-182.

Aspect 46. IL-2Rβ ligand comprises an amino acid sequence selected from any one of SEQ ID NOs: 167-182, wherein the amino acid sequence is on the N-terminus, the C-terminus, or both the N-terminus and the C-terminus 46. An IL-2Rβ ligand according to aspect 45, which ends with the amino acids -GG.

Aspect 47. The IL-2Rβ ligand comprises an amino acid sequence selected from any one of SEQ ID NOS: 167-182, each amino acid independently having the following conservative substitutions: alanine (A), glycine ( G), amino acids with small hydrophobic side chains including proline (P), serine (S), and threonine (T); amino acids with hydroxyl-containing side chains including serine (S) or threonine (T); aspartic acid (D) and amino acids with acidic side chains, including glutamic acid (E); polar, including histidine (H), asparagine (N), glutamine (Q), serine (S), threonine (T), and tyrosine (Y). amino acids with neutral side chains; amino acids with basic side chains including arginine (R), lysine (K), or histidine (H); and isoleucine (I), leucine (L), methionine (M), valine 47. According to any one of aspects 45-46, comprising one or more of amino acids with large hydrophobic side chains, including (V), phenylphenyl (F), tyrosine (Y), or tyrosine (W). IL-2Rβ ligand of.

Aspect 48. the IL-2Rβ ligand comprises the amino acid sequence of formula (2) (SEQ ID NO: 164), the amino acid sequence of formula (2a) (SEQ ID NO: 165), or the amino acid sequence of formula (2b) (SEQ ID NO: 166);
^{-X15 -X16 -X17 -X18 -X19 -X20 -X21 -X22} - (2)
^{-X14 -CX15 -X16 -X17 -X18 -X19 -X20 -X21 -X22 -CX23} - (2a)
^{-X13 -X14 -CX15 -X16 -X17 -X18 -X19 -X20 -X21 -X22 -CX23 -X24} - (2b)
wherein X ¹³ is selected from amino acids, X ¹⁴ is selected from amino acids containing large hydrophobic side chains, X ¹⁵ is selected from amino acids, and X ¹⁶ is selected from amino acids containing large hydrophobic side chains. selected wherein X ²⁷ is selected from amino acids containing small hydrophobic side chains, X ¹⁸ is selected from amino acids, X ¹⁹ is selected from amino acids, and X ²⁰ is selected from amino acids containing large hydrophobic side chains selected wherein ^X21 is selected from amino acids containing small hydrophobic side chains, ^X22 is selected from amino acids, ^X23 is selected from amino acids, and ^X24 is selected from amino acids containing large hydrophobic side chains An IL-2Rβ ligand according to any one of aspects 1-9, which is selected.

Aspect 49. X ¹³ is selected from amino acids, X ¹⁴ is selected from amino acids containing large hydrophobic side chains, X ¹⁵ is selected from amino acids, X ¹⁶ is selected from amino acids containing large hydrophobic side chains, X ¹⁷ is selected from amino acids containing small hydrophobic side chains, X ¹⁸ is selected from amino acids containing polar neutral or acidic side chains, and X ¹⁹ is selected from amino acids containing large hydrophobic or neutral side chains. is selected, X ²⁰ is selected from amino acids containing large hydrophobic side chains, X ²¹ is selected from amino acids containing small hydrophobic side chains, X ²² is selected from amino acids, and X ²³ is selected from amino acids 49. An IL-2Rβ ligand according to aspect 48, wherein ^X24 is selected from amino acids containing large hydrophobic side chains.

Aspect 50. X ¹³ is selected from amino acids, X ¹⁴ is selected from I, L, M, V, F, W, and Y, X ¹⁵ is D, E, I, L, M, V, F, Y, and W, X ¹⁶ is selected from I, L, M, N, V, F, Y, and W; X ¹⁷ is selected from A, ^G , P, S, and T; is selected from H, N, Q, S, T, Y, D, and E, X ¹⁹ is selected from I, L, M, V, F, W, and Y, and X ²⁰ is selected from I, L, M, N, V, F, Y, and W; X ²¹ is selected from A, G, P, S, and T; X ²² is selected from amino acids; X ²³ is an amino acid 49. An IL-2Rβ ligand according to aspect 48, wherein X ²⁴ is selected from I, L, M, V, F, W, and Y.

Aspect 51. 51. An IL-2Rβ ligand according to aspect 50, wherein ^X14 is selected from I and M.

Aspect 52. 52. An IL-2Rβ ligand according to any one of aspects 50-51, wherein ^X16 is V.

Aspect 53. 53. An IL-2Rβ ligand according to any one of aspects 50-52, wherein ^X17 is G.

Aspect 54. 54. An IL-2Rβ ligand according to any one of aspects 50-53, wherein ^X18 is selected from D and Q.

Aspect 55. 55. An IL-2Rβ ligand according to any one of aspects 50-54, wherein ^X20 is W.

Aspect 56. 56. An IL-2Rβ ligand according to any one of aspects 50-55, wherein ^X21 is P.

Aspect 57. 57. An IL-2Rβ ligand according to any one of aspects 50-56, wherein ^X23 is selected from F, I, L and V.

Aspect 58. X ¹³ is selected from amino acids, X ¹⁴ is selected from I and M, X ¹⁵ is selected from amino acids, X ¹⁶ is V, X ¹⁷ is G, X ¹⁸ is D and Q, X ¹⁹ is selected from I, L, M, V, F, W, and Y, X ²⁰ is W, X ²¹ is P, and X ²² is from an amino acid 51. An IL-2Rβ ligand according to aspect 50, wherein ^X23 is selected from amino acids and ^X24 is selected from F, I, L, and V.

Aspect 59. the IL-2Rβ ligand comprises the amino acid sequence of formula (3) (SEQ ID NO: 183) or the amino acid sequence of formula (3a) (SEQ ID NO: 184);
^{-X26 -X27 -X28 -X29 -X30} -X31 ^{-X32 -X33 -X34 -X35} - ( ³ )
^{-X25 -CX26 -X27 -X28 -X29 -X30 -X31 -X32 -X33 -X34 -X35 -CX36} - (3a)
wherein X ²⁵ is selected from amino acids, X ²⁶ is selected from amino acids, X ²⁷ is selected from I and V, X ²⁸ is selected from G, X ²⁹ is D, E, and N, X ³⁰ is selected from F, L, and Y, X ³¹ is selected from F, I, and V, X ³² is selected from D and Q, X ³³ is from amino acids 10. An IL-2Rβ ligand according to any one of aspects 1-9, wherein X ³⁴ is selected from amino acids, X ³⁵ is selected from amino acids, and X ³⁶ is selected from amino acids.

Aspect 60. X ²⁵ is selected from L, S, T, and Y ^; X ²⁶ is selected from ^H and Q; X ²⁷ is selected from I and V; D, E, and N; X ³⁰ is selected from F, L, and Y; X ³¹ is selected from F, I, and V; X ³² is selected from D and Q; ³³ is selected from D, L, and W; X ³⁴ is selected from G, L, ^{and T; X 35 is} selected from D, I, and S; 60. An IL-2Rβ ligand according to aspect 59, wherein the IL-2Rβ ligand is

Aspect 61. the IL-2Rβ ligand comprises the amino acid sequence of formula (3) (SEQ ID NO: 183) or the amino acid sequence of formula (3a) (SEQ ID NO: 184);
^{-X26 -X27 -X28 -X29 -X30 -X31 -X32 -X33 -X34 -X35} - ( ³ )
^{-X25 -CX26 -X27 -X28 -X29 -X30 -X31 -X32 -X33 -X34 -X35 -CX36} - (3a)
wherein X ²⁵ is selected from amino acids, X ²⁶ is selected from amino acids, X ²⁷ is selected from amino acids containing large hydrophobic side chains, and X ²⁸ is selected from amino acids containing small hydrophobic side chains. selected wherein X ²⁹ is selected from amino acids comprising an acidic side chain or a polar neutral side chain, X ³⁰ is selected from amino acids, X ³¹ is selected from amino acids, X ³² is a polar neutral side chain or selected from amino acids comprising an acidic side chain, X ³³ is selected from amino acids, X ³⁴ is selected from amino acids, X ³⁵ is selected from amino acids, and X ³⁶ is selected from amino acids, aspect 1 10. IL-2Rβ ligand according to any one of 1-9.

Aspect 62. X ²⁵ is selected from amino acids, X ²⁶ is selected from amino acids, X ²⁷ is selected from amino acids containing large hydrophobic side chains, X ²⁸ is selected from amino acids containing small hydrophobic side chains, X ²⁹ is selected from amino acids containing acidic or polar neutral side chains, X ³⁰ is selected from amino acids containing large hydrophobic side chains, and X ³¹ is selected from amino acids containing large hydrophobic side chains. X ³² is selected from amino acids comprising a polar neutral side chain or an acidic side chain, X ³³ is selected from amino acids, X ³⁴ is selected from amino acids, X ³⁵ is selected from amino acids, X 62. An IL-2Rβ ligand according to aspect 61, wherein ³⁶ are selected from amino acids.

Aspect 63. the IL-2Rβ ligand comprises the amino acid sequence of formula (3) (SEQ ID NO: 183) or the amino acid sequence of formula (3a) (SEQ ID NO: 184);
^{-X26 -X27 -X28 -X29 -X30} -X31 ^{-X32 -X33 -X34 -X35} - ( ³ )
^{-X25 -CX26 -X27 -X28 -X29 -X30 -X31 -X32 -X33 -X34 -X35 -CX36} - (3a)
wherein X ²⁵ is selected from amino acids; X ²⁶ is selected from amino acids; X ²⁷ is selected from I, ^L , M, V, F, Y, and W; , P, S, and T; X ²⁹ is selected from D, E, H, N, Q, S, T, and Y; X ³⁰ is selected from I, L, M, V, F, Y , and W, X ³¹ is selected from I, L, M, V, F, Y, and W, and X ³² is selected from D, E, H, N, Q, T, and Y , X ³³ is selected from amino acids, X ³⁴ is selected from amino acids, X ³⁵ is selected from amino acids, and X ³⁶ is selected from amino acids. IL-2Rβ ligand.

Aspect 64. 64. An IL-2Rβ ligand according to aspect 63, wherein ^X27 is selected from V and I.

Aspect 65. 65. An IL-2Rβ ligand according to any one of aspects 63-64, wherein ^X28 is G.

Aspect 67. 66. An IL-2Rβ ligand according to any one of aspects 63-65, wherein ^X29 is selected from D and E.

Aspect 68. 67. An IL-2Rβ ligand according to any one of aspects 63-66, wherein ^X30 is selected from V, L, F and Y.

Aspect 69. 68. An IL-2Rβ ligand according to any one of aspects 63-67, wherein ^X31 is selected from I, V and F.

Embodiment 69a. 69. An IL-2Rβ ligand according to any one of aspects 63-68, wherein ^X32 is selected from Q and D.

Aspect 70. X ²⁵ is selected from amino acids, X ²⁶ is selected from amino acids, X ²⁷ is selected from V and I, X ²⁸ is selected from G, X ²⁹ is selected from D and E, X ³⁰ is selected from V, L, F, and Y; X ³¹ is selected from I, V, and F; X ³² is selected from Q and D; X ³³ is selected from amino acids; ^64. An IL-2Rβ ligand according to aspect 63, wherein 34 is selected from amino acids, X ³⁵ is selected from amino acids and X ³⁶ is selected from amino acids.

Aspect 71. An IL-2Rβ ligand according to any one of aspects 1-9, wherein the IL-2Rβ ligand comprises an amino acid sequence selected from any one of SEQ ID NOs:185-193.

Aspect 72. IL-2Rβ ligand comprises an amino acid sequence selected from any one of SEQ ID NOs: 185-193, wherein the amino acid sequence is on the N-terminus, the C-terminus, or both the N-terminus and the C-terminus 72. An IL-2Rβ ligand according to aspect 71, which ends with the amino acids -GG.

Aspect 73. The IL-2Rβ ligand comprises an amino acid sequence selected from any one of SEQ ID NOS: 185-193, each amino acid independently having the following conservative substitutions: alanine (A), glycine ( G), amino acids with small hydrophobic side chains including proline (P), serine (S), and threonine (T); amino acids with hydroxyl-containing side chains including serine (S) or threonine (T); aspartic acid (D) and amino acids with acidic side chains, including glutamic acid (E); polar, including histidine (H), asparagine (N), glutamine (Q), serine (S), threonine (T), and tyrosine (Y). amino acids with neutral side chains; amino acids with basic side chains including arginine (R), lysine (K), or histidine (H); and isoleucine (I), leucine (L), methionine (M), valine 73. According to any one of aspects 71-72, comprising one or more of amino acids with large hydrophobic side chains, including (V), phenylphenyl (F), tyrosine (Y), or tyrosine (W). IL-2Rβ ligand of.

Aspect 74. An IL-2Rγc ligand, wherein the IL-2Rγc ligand exhibits a binding affinity for the human IL-2Rγc subunit of less than 100 μM.

Aspect 75. 75. An IL-2Rγc ligand according to aspect 74, wherein the IL-2Rγc ligand comprises 5-30 amino acids.

Aspect 76. 76. An IL-2Rγc ligand according to any one of aspects 74-75, wherein the IL-2Rγc ligand exhibits a binding affinity for human IL-2Rγc subunits ranging from 1 pM to 100 μM.

Aspect 77. 76. An IL-2Rγc ligand according to any one of aspects 74-75, wherein the IL-2Rγc ligand exhibits a binding affinity for human IL-2Rγc subunits ranging from 0.1 μM to 50 μM.

Aspect 78. 76. An IL-2Rγc ligand according to any one of aspects 74-75, wherein the IL-2Rγc ligand exhibits a binding affinity for human IL-2Rγc subunits of less than 100 μM.

Aspect 79. 79. An IL-2Rγc ligand according to any one of aspects 74-78, wherein the IL-2Rγc ligand exhibits a binding affinity of less than 100 μM for each of the human IL-2Rβ subunit and the human IL-2Rγc subunit.

Aspect 80. 80. An IL-2Rγc ligand according to any one of aspects 74-79, wherein the IL-2Rγc ligand exhibits binding affinity for the human IL-2Rα (CD25) subunit.

Aspect 81. 81. IL-2Rγc according to any one of aspects 74 to 80, wherein the IL-2Rγc ligand is at least 10 times the binding affinity for the human IL-2Rγc subunit of the IL-2Rγc ligand for the human IL-2Rγc subunit. ligand.

Aspect 82. the IL-2Rγc ligand comprises the amino acid sequence of formula (4) (SEQ ID NO: 194) or the amino acid sequence of formula (4a) (SEQ ID NO: 195);
^{-X53 -X54 -X55 -X56 -X57 -X58 -X59 -X60} - (4)
^{-X51 -X52 -CX53 -X54 -X55 -X56 -X57 -X58 -X59 -X60 -CX61 -X62} - (4a)
wherein X ⁵¹ is selected from G, I, K, L, Q, R, T, and V; and X ⁵² is selected from A, E, I, L, R, S, T, V, and W; is selected, X ⁵³ is selected from D, E, F, N, Q, S, and T; X ⁵⁴ is selected from D, E, I, M, N, Q, R, and S; ⁵⁵ is selected from D, ^E , F, S, T, W, and Y; X ⁵⁶ is selected from D, E, F, G, L, M, N, Q, and Y; , E, G, and N, X ⁵⁸ is selected from I, P, T, and V, X ⁵⁹ is selected from I, L, M, S, T, and V, and X ⁶⁰ is selected from , F, I, and L, X ⁶¹ is selected from F, T, and W, and X ⁶² is selected from A, E, G, L, N, P, Q, S, and W 82. An IL-2Rγc ligand according to any one of aspects 74-81.

Aspect 83. 83. An IL-2Rγc ligand according to aspect 82, wherein ^X51 is selected from I, L, and V.

Aspect 84. 84. An IL-2Rγc ligand according to any one of aspects 82-83, wherein ^X52 is selected from S and T.

Aspect 85. 85. An IL-2Rγc ligand according to any one of aspects 82-84, wherein ^X53 is selected from D, E, N, and Q.

Aspect 86. 86. An IL-2Rγc ligand according to any one of aspects 82-85, wherein ^X54 is selected from D, E, N, and Q.

Aspect 87. 87. An IL-2Rγc ligand according to any one of aspects 82-86, wherein ^X55 is selected from F, W and Y.

Aspect 88. 88. An IL-2Rγc ligand according to any one of aspects 82-87, wherein ^X56 is selected from D, E, N, and Q.

Aspect 89. 89. An IL-2Rγc ligand according to any one of aspects 82-88, wherein ^X57 is G.

Aspect 90. 89. An IL-2Rγc ligand according to any one of aspects 82-89, wherein ^X58 is selected from I and V.

Aspect 91. 91. An IL-2Rγc ligand according to any one of aspects 82-90, wherein ^X59 is selected from I, L, M and V.

Aspect 92. 92. An IL-2Rγc ligand according to any one of aspects 82-91, wherein ^X60 is selected from F, I, and L.

Aspect 93. An IL-2Rγc ligand according to any one of aspects 82-92, wherein ^X61 is W.

Aspect 94. 94. An IL-2Rγc ligand according to any one of aspects 82-93, wherein ^X62 is selected from N and Q.

Aspect 95. X ⁵¹ is selected from I, L, and V; X ⁵² is selected from S ^and T; X ⁵³ is selected from D, E, N, and Q; , X ⁵⁵ is selected from F, W, and Y, X ⁵⁶ is selected from D, E, N, and Q, X ⁵⁷ is selected from G, and X ⁵⁸ is selected from I and V , X ⁵⁹ is selected from I, L, M, and V, X ⁶⁰ is selected from F, I, and L, X ⁶¹ is W, and X ⁶² is selected from N and Q 83. An IL-2Rγc ligand according to aspect 82, wherein the IL-2Rγc ligand is

Aspect 96. 82. An IL-2Rγc ligand according to any one of aspects 74-81, wherein the IL-2Rγc ligand comprises an amino acid sequence selected from any one of SEQ ID NO:196-SEQ ID NO:210.

Aspect 97. IL-2Rγc ligand comprises an amino acid sequence selected from any one of SEQ ID NOs: 196-210, wherein the amino acid sequence is on the N-terminus, the C-terminus, or both the N-terminus and the C-terminus 97. An IL-2Rγc ligand according to aspect 96, which ends with the amino acids -GG.

Aspect 98. The IL-2Rβ ligand comprises an amino acid sequence selected from any one of SEQ ID NOs: 196-210, each amino acid independently having the following conservative substitutions: alanine (A), glycine ( G), amino acids with small hydrophobic side chains including proline (P), serine (S), and threonine (T); amino acids with hydroxyl-containing side chains including serine (S) or threonine (T); aspartic acid (D) and amino acids with acidic side chains, including glutamic acid (E); polar, including histidine (H), asparagine (N), glutamine (Q), serine (S), threonine (T), and tyrosine (Y). amino acids with neutral side chains; amino acids with basic side chains including arginine (R), lysine (K), or histidine (H); and isoleucine (I), leucine (L), methionine (M), valine. 98. According to any one of aspects 96-97, comprising one or more of amino acids with large hydrophobic side chains, including (V), phenylphenyl (F), tyrosine (Y), or tyrosine (W). IL-2Rβ ligand of.

Aspect 99. the IL-2Rγc ligand comprises the amino acid sequence of formula (4) (SEQ ID NO: 194) or the amino acid sequence of formula (4a) (SEQ ID NO: 195);
^{-X53 -X54 -X55 -X56 -X57 -X58 -X59 -X60} - (4)
^{-X51 -X52 -CX53 -X54 -X55 -X56 -X57 -X58 -X59 -X60 -CX61 -X62} - (4a)
wherein X ⁵¹ is selected from amino acids, X ⁵² is selected from amino acids, X ⁵³ is selected from amino acids containing polar neutral side chains or acidic side chains, X ⁵⁴ is selected from polar neutral side chains or selected from amino acids containing acidic side chains, X ⁵⁵ is selected from amino acids, X ⁵⁶ is selected from amino acids, X ⁵⁷ is selected from amino acids containing small hydrophobic side chains, X ⁵⁸ is selected from large is selected from amino acids containing a hydrophobic side chain, X ⁵⁹ is selected from amino acids containing a large hydrophobic side chain, X ⁶⁰ is selected from amino acids containing a large hydrophobic side chain, X ⁶¹ is selected from amino acids containing a large hydrophobic side chain 82. An IL-2Rγc ligand according to any one of aspects 74-81, wherein ^X62 is selected from amino acids comprising side chains, and wherein X62 is selected from amino acids.

Embodiment 100. ^X51 is selected from amino acids containing large hydrophobic and basic side chains, ^X52 is selected from amino acids containing hydroxyl-containing and large hydrophobic side chains, and ^X53 is polar neutral. X ⁵⁴ is selected from amino acids containing a polar neutral side chain or an acidic side chain, X ⁵⁵ is selected from amino acids containing a large hydrophobic side chain, X ⁵⁶ is selected from amino acids containing polar neutral or acidic side chains, X ⁵⁷ is selected from amino acids containing small hydrophobic side chains, X ⁵⁸ is selected from amino acids containing large hydrophobic side chains, X ⁵⁹ is selected from amino acids with large hydrophobic side chains, X ⁶⁰ is selected from amino acids with large hydrophobic side chains, X ⁶¹ is selected from amino acids with large hydrophobic side chains, X ⁶² is selected from amino acids containing polar neutral side chains.

Aspect 101. X ⁵¹ is selected from R, K, H, F, I, L, M, V, Y, and W, and X ⁵² is S, T, F, I, L, M, V, Y, and W with X ⁵³ selected from D, E, H, N, Q, S, T, and Y; X ⁵⁴ selected from D, E, H, N, Q, S, T, and Y , X ⁵⁵ is selected from F, I, L, M, V, Y, ^and W; X ⁵⁶ is selected from D, E, H, N, Q, S, T, and Y; is selected from A, G, P, S, and T; X ⁵⁸ is selected from F, I, L, M, V, Y, and W; X ⁵⁹ is selected from F, I, L, M, V, Y, and W; X ⁶⁰ is selected from F, I, L, M, V, Y, and W; X ⁶¹ is F, I, L, M, V, Y, and W and X ⁶² is selected from H, N, Q, S, and Y. IL-2Rγc ligand according to aspect 99.

Aspect 102. 102. An IL-2Rγc ligand according to aspect 101, wherein ^X51 is selected from I, L, and V.

Aspect 103. 103. An IL-2Rγc ligand according to any one of aspects 101-102, wherein ^X52 is selected from S and T.

Aspect 104. 103. An IL-2Rγc ligand according to any one of aspects 101-103, wherein ^X53 is selected from D, E, and Q.

Aspect 105. 105. An IL-2Rγc ligand according to any one of aspects 101-104, wherein ^X54 is selected from D, E, and N.

Aspect 106. 106. An IL-2Rγc ligand according to any one of aspects 101-105, wherein ^X55 is selected from F, Y and W.

Aspect 107. 107. An IL-2Rγc ligand according to any one of aspects 101-106, wherein ^X56 is selected from D, E, N, and Q.

Aspect 108. 107. An IL-2Rγc ligand according to any one of aspects 101-107, wherein ^X57 is G.

Aspect 109. 109. An IL-2Rγc ligand according to any one of aspects 101-108, wherein ^X58 is selected from I and V.

Aspect 110. 109. An IL-2Rγc ligand according to any one of aspects 101-109, wherein ^X59 is selected from I, L, M, and V.

Aspect 111. 111. An IL-2Rγc ligand according to any one of aspects 101-110, wherein ^X60 is selected from F, I, and L.

Aspect 112. An IL-2Rγc ligand according to any one of aspects 101-111, wherein ^X61 is W.

Aspect 113. 113. An IL-2Rγc ligand according to any one of aspects 101-112, wherein ^X62 is selected from N and Q.

Aspect 114. X ⁵¹ is selected from I, L, and V; X ⁵² is selected from S ^and T; X ⁵³ is selected from D, E, and Q; X ⁵⁵ is selected from F, Y, and W; X ⁵⁶ is selected from D ^{, E, N, and Q; X 57 is} selected from G; X ⁵⁹ is selected from I, L, M, and V, X ⁶⁰ is selected from F, I, and L, X ⁶¹ is W, and X ⁶² is selected from N and Q 102. An IL-2Rγc ligand according to aspect 101, wherein the IL-2Rγc ligand is

Aspect 115. the IL-2Rγc ligand comprises the amino acid sequence of Formula (5) (SEQ ID NO:211) or Formula (5a) (SEQ ID NO:212);
^{-X73 -X74 -X75 -X76 -X77 -X78 -X79 -X80} - (5)
^{-X71 -X72 -CX73 -X74 -X75 -X76 -X77 -X78 -X79 -X80 -CX81 -X82} - (5a)
wherein X ⁷¹ is selected from I, L, P, Q, R, T, and V; X ⁷² is selected from A, D, E, I, M, R, T, and V; ⁷³ is selected from E, M, N, Q, S, T, V, W, and Y, and X ⁷⁴ is selected from D, E, F, G, I, M, R, S, T, and V X ⁷⁵ is selected from F, W, and Y; X ⁷⁶ is selected from D, E, L, N, Q, and S; ^{X 77} is selected from G; is selected from I, M, and V; ^{X 79} is selected from D, E, N, Q, and R; X ⁸⁰ is selected from F, I, and L; any one of aspects 74 through 81, wherein X 82 is selected from R, T, W, and Y, and X ⁸² is selected from A, F, H, I, L, N, P, Q, S, and W An IL-2Rγc ligand as described in .

Aspect 116. 116. An IL-2Rγc ligand according to aspect 115, wherein ^X71 is selected from I, L, and V.

Aspect 117. 117. An IL-2Rγc ligand according to any one of aspects 115-116, wherein ^X72 is selected from A, D, E, I, M, and V.

Aspect 118. 118. An IL-2Rγc ligand according to any one of aspects 115-117, wherein ^X73 is selected from E, Q and N.

Aspect 119. 119. An IL-2Rγc ligand according to any one of aspects 115-118, wherein ^X74 is selected from D and E.

Aspect 120. 120. An IL-2Rγc ligand according to any one of aspects 115-119, wherein ^X75 is selected from F, W, and Y.

Aspect 121. 121. An IL-2Rγc ligand according to any one of aspects 115-120, wherein ^X76 is selected from D, E, L, N, and Q.

Aspect 122. 122. An IL-2Rγc ligand according to any one of aspects 115-121, wherein ^X77 is G.

Aspect 123. 123. An IL-2Rγc ligand according to any one of aspects 115-122, wherein ^X78 is selected from I, M, and V.

Aspect 124. 124. An IL-2Rγc ligand according to any one of aspects 115-123, wherein ^X79 is selected from D, E, Q and R.

Aspect 125. 125. An IL-2Rγc ligand according to any one of aspects 115-124, wherein ^X80 is selected from F, I, and L.

Aspect 126. 126. An IL-2Rγc ligand according to any one of aspects 115-125, wherein ^X81 is W.

Aspect 127. 127. An IL-2Rγc ligand according to any one of aspects 115-126, wherein ^X82 is selected from N and Q.

Aspect 128. X ⁷¹ is selected from I, L, and V; X ⁷² is selected from A, D, E, I, M, and V; X ⁷³ is selected from E, Q, and ^N ; is selected from D and E, X ⁷⁵ is selected from F, W, and Y, X ⁷⁶ is selected from D, E, L, N, and Q, X ⁷⁷ is G, X ⁷⁸ is selected from I, M, and V; ^{X 79} is selected from D, E, Q, and R; X ⁸⁰ is selected from F, I, and L; 116. An IL-2Rγc ligand according to aspect 115, wherein X ⁸² is selected from N and Q.

Aspect 129. the IL-2Rγc ligand comprises the amino acid sequence of Formula (5) (SEQ ID NO:211) or Formula (5a) (SEQ ID NO:212);
^{-X73 -X74 -X75 -X76 -X77 -X78 -X79 -X80} - (5)
^{-X71 -X72 -CX73 -X74 -X75 -X76 -X77 -X78 -X79 -X80 -CX81 -X82} - (5a)
wherein X ⁶¹ is selected from amino acids, X ⁶² is selected from amino acids, X ⁶³ is selected from amino acids, X ⁶⁴ is selected from amino acids, and X ⁶⁵ contains a large hydrophobic side chain is selected from amino acids, X ⁶⁶ is selected from amino acids, X ⁶⁷ is selected from small hydrophobic side chains, X ⁶⁸ is selected from amino acids containing large hydrophobic side chains, X ⁶⁹ is selected from basic side chains X ⁷⁰ is selected from amino acids containing large hydrophobic side chains, and X ⁷¹ is selected from amino acids containing large hydrophobic side chains. , X ⁷² are selected from amino acids.

Aspect 130. X ⁷¹ is selected from amino acids containing large hydrophobic side chains, X ⁷² is selected from amino acids containing acidic or large hydrophobic side chains, X ⁷³ is selected from acidic side chains, hydroxyl-containing side chains, or is selected from amino acids containing a polar neutral side chain, X ⁷⁴ is selected from amino acids containing an acidic side chain, a hydroxyl-containing side chain, or a large hydrophobic side chain, and X ⁷⁵ is an amino acid containing a large hydrophobic side chain wherein X ⁷⁶ is selected from an amino acid containing an acidic, hydroxyl-containing or polar neutral side chain, X ⁷⁷ is selected from a small hydrophobic side chain, X ⁷⁸ is a large hydrophobic is selected from amino acids containing side chains, X ⁷⁹ is selected from amino acids containing basic, acidic, or polar neutral side chains, and X ⁸⁰ is selected from amino acids containing large hydrophobic side chains; 130. An IL-2Rγc ligand according to aspect 129, wherein X ⁸¹ is selected from amino acids containing large hydrophobic side chains and X ⁸² is selected from amino acids containing polar neutral side chains.

Aspect 131. X ⁷¹ is selected from F, I, L, M, V, Y, and W; X ⁷² is selected from D, E, F, I, L, M, V ^, Y, and W; is selected from D, E, S, T, H, N, Q, S, T, and Y, and X ⁷⁴ is selected from D, E, S, T, F, I, L, M, V, Y, and W, X ⁷⁵ is selected from F, I, L, M, V, Y, and W, X ⁷⁶ is selected from D, E, S, T, H, N, Q, S, T, and Y; X ⁷⁷ is selected from A, G, P, S, and T; X ⁷⁸ is selected from F, I, L, M, V, Y, and ^W ; R, K, H, D, E, H, N, Q, S, T, and Y; X ⁸⁰ is selected from F, I, L, M, V, Y, and W; X ⁸¹ is selected from F, I, L, M, V, Y, and W; and X ⁸² is selected from H, N, Q, S, T, and Y. .

Aspect 132. 132. An IL-2Rγc ligand according to aspect 131, wherein ^X71 is selected from I, L, and V.

Aspect 133. 133. An IL-2Rγc ligand according to any one of aspects 131-132, wherein ^X72 is selected from D, E, I, M, and V.

Aspect 134. 134. An IL-2Rγc ligand according to any one of aspects 131-133, wherein ^X73 is selected from E, N, and Q.

Aspect 135. 135. An IL-2Rγc ligand according to any one of aspects 131-134, wherein ^X74 is selected from D and E.

Aspect 136. 136. An IL-2Rγc ligand according to any one of aspects 131-135, wherein ^X75 is selected from F, W, and Y.

Aspect 137. 137. An IL-2Rγc ligand according to any one of aspects 131-136, wherein ^X76 is selected from D, E, and N.

Aspect 138. 138. An IL-2Rγc ligand according to any one of aspects 131-137, wherein ^X77 is selected from G.

Aspect 139. 139. An IL-2Rγc ligand according to any one of aspects 131-138, wherein ^X78 is selected from I, M, and V.

Aspect 140. 139. An IL-2Rγc ligand according to any one of aspects 131-139, wherein ^X79 is selected from D, E, N, Q, and R.

Aspect 141. 141. An IL-2Rγc ligand according to any one of aspects 131-140, wherein ^X80 is selected from F, I, and L.

Aspect 142. 142. An IL-2Rγc ligand according to any one of aspects 131-141, wherein ^X81 is W.

Aspect 143. 143. An IL-2Rγc ligand according to any one of aspects 131-142, wherein ^X82 is selected from N and Q.

Aspect 144. X ⁷¹ is selected from I, L, and V; X ⁷² is selected from D, E, I, ^M , and V; X ⁷³ is selected from E, N, and Q; D and E, X ⁷⁵ is selected from F, W, and Y, X ⁷⁶ is selected from D, E, and N, X ⁷⁷ is selected from G, X ⁷⁸ is selected from I, M, and V; X ⁷⁹ is selected from D, E, N, Q, and R; X ⁸⁰ is selected from F, I, and L; X ⁸¹ is ^W ; is selected from N and Q. IL-2Rγc ligand according to aspect 131.

Aspect 145. 82. An IL-2Rγc ligand according to any one of aspects 74-81, wherein the IL-2Rγc ligand comprises an amino acid sequence selected from any one of SEQ ID NO:213-SEQ ID NO:233.

Aspect 146. The IL-2Rγc ligand comprises an amino acid sequence selected from any one of SEQ ID NOs:213-233, wherein the amino acid sequence is on the N-terminus, the C-terminus, or both the N-terminus and the C-terminus 146. An IL-2Rγc ligand according to aspect 145, which ends with amino acids -GG of.

Aspect 147. The IL-2Rβ ligand comprises an amino acid sequence selected from any one of SEQ ID NOS:213-233, each amino acid independently having the following conservative substitutions: alanine (A), glycine ( G), amino acids with small hydrophobic side chains including proline (P), serine (S), and threonine (T); amino acids with hydroxyl-containing side chains including serine (S) or threonine (T); aspartic acid (D) and amino acids with acidic side chains, including glutamic acid (E); polar, including histidine (H), asparagine (N), glutamine (Q), serine (S), threonine (T), and tyrosine (Y). amino acids with neutral side chains; amino acids with basic side chains including arginine (R), lysine (K), or histidine (H); and isoleucine (I), leucine (L), methionine (M), valine 147. according to any one of aspects 145-146, comprising one or more of amino acids with large hydrophobic side chains, including (V), phenylphenyl (F), tyrosine (Y), or tyrosine (W) IL-2Rβ ligand of.

Aspect 148. the IL-2Rγc ligand comprises the amino acid sequence of Formula (6) (SEQ ID NO:234) or Formula (6a) (SEQ ID NO:235);
^{-X93 -X94 -X95 -X96 -X97 -X98 -X99 -X100 -X101} - (6)
^{-X91 -X92 -X93 -X94 -X95 -X96 -X97 -X98 -X9 -X100 -X101 -X102 -X103} - (6a)
wherein X ⁹¹ is selected from C, D, E, and L; X ⁹² is selected from C, L, M, R, S, V, and W; X ⁹³ is selected from C, D, F , P, and R, X ⁹⁴ is selected from A, D, L, Q, S, and W, X ⁹⁵ is selected from D, E, F, L, and V, and X ⁹⁶ is selected from , A, D, E, F, G, K, Q, and S, X ⁹⁷ is selected from E, L, M, and W, X ⁹⁸ is G, I, L, W, and Y, X ⁹⁹ is selected from E, I, R, T, and V, X ¹⁰⁰ is W, and X ¹⁰¹ is selected from C, A, I, L, P, and V , X ¹⁰² is selected from C, D, G, H, and X ¹⁰³ is selected from C, D, E, H, S, and T. -2Rγc ligand.

Aspect 149. 149. The IL-2Rγc ligand of embodiment 148, wherein ^X91 is selected from D and E.

Aspect 150. 149. An IL-2Rγc ligand according to any one of aspects 148-149, wherein ^X92 is selected from L, M, R, S, V, and W.

Aspect 151. 151. An IL-2Rγc ligand according to any one of aspects 148-150, wherein ^X93 is selected from D and F.

Aspect 152. 152. An IL-2Rγc ligand according to any one of aspects 148-151, wherein ^X94 is S.

Aspect 153. 153. An IL-2Rγc ligand according to any one of aspects 148-152, wherein ^X95 is selected from D and E.

Aspect 154. 154. An IL-2Rγc ligand according to any one of aspects 148-153, wherein ^X96 is selected from D and E.

Aspect 155. 155. An IL-2Rγc ligand according to any one of aspects 148-154, wherein ^X97 is selected from L, M and W.

Aspect 156. 156. An IL-2Rγc ligand according to any one of aspects 148-155, wherein X ⁹⁸ is G.

Aspect 157. 157. An IL-2Rγc ligand according to any one of aspects 148-156, wherein ^X99 is E.

Aspect 158. 158. An IL-2Rγc ligand according to any one of aspects 148-157, wherein X ¹⁰⁰ is W.

Aspect 159. 159. An IL-2Rγc ligand according to any one of aspects 148-158, wherein X ¹⁰¹ is selected from I, L, and V.

Aspect 160. 159. An IL-2Rγc ligand according to any one of aspects 148-159, wherein X ¹⁰² is selected from D and G.

Aspect 161. 161. An IL-2Rγc ligand according to any one of aspects 148-160, wherein X ¹⁰³ is selected from S and T.

Aspect 162. X ⁹¹ is selected from D and E; X ⁹² is selected from L, M, R, S, V, and W; X ⁹³ is selected from D and F; X ⁹⁴ is S; X ⁹⁵ is selected from D and E, X ⁹⁶ is selected from D and E, X ⁹⁷ is selected from L, M and W, X ⁹⁸ is G and X ⁹⁹ is E 148, wherein X ¹⁰⁰ is W, X ¹⁰¹ is selected from I, L, and V, X ¹⁰² is selected from D and G, and X ¹⁰³ is selected from S and T IL-2Rγc ligands as described.

Aspect 163. 82. An IL-2Rγc ligand according to any one of aspects 74-81, wherein the IL-2Rγc ligand comprises an amino acid sequence selected from any one of SEQ ID NOs:236-245.

Aspect 164. the IL-2Rγc ligand comprises an amino acid sequence selected from any one of SEQ ID NOs:236-245, wherein the amino acid sequence is on the N-terminus, the C-terminus, or both the N-terminus and the C-terminus 164. An IL-2Rγc ligand according to aspect 163, which ends with the amino acids -GG.

Aspect 165. The IL-2Rβ ligand comprises an amino acid sequence selected from any one of SEQ ID NOs:236-245, each amino acid independently having the following conservative substitutions: alanine (A), glycine ( G), amino acids with small hydrophobic side chains including proline (P), serine (S), and threonine (T); amino acids with hydroxyl-containing side chains including serine (S) or threonine (T); aspartic acid (D) and amino acids with acidic side chains, including glutamic acid (E); polar, including histidine (H), asparagine (N), glutamine (Q), serine (S), threonine (T), and tyrosine (Y). amino acids with neutral side chains; amino acids with basic side chains including arginine (R), lysine (K), or histidine (H); and isoleucine (I), leucine (L), methionine (M), valine 164. According to any one of aspects 163-164, comprising one or more of amino acids with large hydrophobic side chains, including (V), phenylphenyl (F), tyrosine (Y), or tyrosine (W). IL-2Rβ ligand of.

Aspect 166. the IL-2Rγc ligand comprises the amino acid sequence of Formula (6) (SEQ ID NO:234) or Formula (6a) (SEQ ID NO:235);
^{-X93 -X94 -X95 -X96 -X97 -X98 -X99 -X100 -X101} - (6)
^{-X91 -X92 -X93 -X94 -X95 -X96 -X97 -X98 -X9 -X100 -X101 -X102 -X103} - (6a)
wherein X ⁹¹ is selected from amino acids containing acidic side chains or cysteine, X ⁹² is selected from amino acids, X ⁹³ is selected from amino acids containing acidic side chains or large hydrophobic side chains, and X ⁹⁴ is selected from amino acids, X ⁹⁵ is selected from amino acids, X ⁹⁶ is selected from amino acids, X ⁹⁷ is selected from amino acids containing large hydrophobic side chains, X ⁹⁸ is selected from small hydrophobic side chains or selected from amino acids with large hydrophobic side chains, X ⁹⁹ is selected from amino acids, X ¹⁰⁰ is selected from amino acids with large hydrophobic side chains, and X ¹⁰¹ is selected from amino acids with large hydrophobic side chains. wherein X ¹⁰² is selected from amino acids containing small hydrophobic or acidic side chains or cysteine, and X ¹⁰³ is selected from amino acids containing acidic or hydroxyl-containing side chains or cysteine, 81. The IL-2Rγc ligand of any one of 81.

Aspect 167. X ⁹¹ is selected from amino acids containing acidic side chains, X ⁹² is selected from amino acids, X ⁹³ is selected from amino acids containing acidic side chains or large hydrophobic side chains, and X ⁹⁴ is selected from acidic side chains. or selected from amino acids containing hydroxyl-containing side chains, X ⁹⁵ is selected from amino acids containing acidic side chains, X ⁹⁶ is selected from amino acids, and X ⁹⁷ is selected from amino acids containing large hydrophobic side chains. , X ⁹⁸ is selected from amino acids containing small or large hydrophobic side chains, X ⁹⁹ is selected from amino acids containing acidic or large hydrophobic side chains, and X ¹⁰⁰ is selected from amino acids containing large hydrophobic side chains. is selected from amino acids containing side chains, X ¹⁰¹ is selected from amino acids containing large hydrophobic side chains, X ¹⁰² is selected from amino acids containing small hydrophobic or acidic side chains, and X ¹⁰³ is acidic 167. An IL-2Rγc ligand according to aspect 166, selected from amino acids containing side chains or hydroxyl-containing side chains.

Aspect 168. X ⁹¹ is selected from D and E, X ⁹² is selected from amino acids, X ⁹³ is selected from D, E, F, I, L, M, V, Y, and W, X ⁹⁴ is D, E, S, and T; X ⁹⁵ is selected from D and E; X ⁹⁶ is selected from amino acids; X ⁹⁷ is F, I, L, M, V, Y, and W X ⁹⁸ is selected from A, G, P, S, T, F, I, L, M, V, Y, and W; X ⁹⁹ is selected from D, E, F, I, L, M, V, Y, and W; X ¹⁰⁰ is selected from F, I, L, M, V, Y, and W; X ¹⁰¹ is selected from F, I, L, M, V, Y, and W, X ¹⁰² is selected from D, E, A, G, P, S, and T, and X ¹⁰³ is selected from D, E, S, and T. IL-2Rγc ligand.

Aspect 169. 169. An IL-2Rγc ligand according to embodiment 168, wherein ^X91 is selected from D and E.

Aspect 170. 169. An IL-2Rγc ligand according to any one of aspects 168-169, wherein ^X92 is selected from amino acids.

Aspect 171. 171. An IL-2Rγc ligand according to any one of aspects 168-170, wherein ^X93 is selected from D and F.

Aspect 172. 172. An IL-2Rγc ligand according to any one of aspects 168-171, wherein ^X94 is S.

Aspect 173. 173. An IL-2Rγc ligand according to any one of aspects 168-172, wherein ^X95 is selected from D and E.

Aspect 174. 174. An IL-2Rγc ligand according to any one of aspects 168-173, wherein ^X96 is selected from amino acids.

Aspect 175. 175. An IL-2Rγc ligand according to any one of aspects 168-174, wherein ^X97 is selected from L, M and W.

Aspect 176. 176. An IL-2Rγc ligand according to any one of aspects 168-175, wherein X ⁹⁸ is G.

Aspect 177. 177. An IL-2Rγc ligand according to any one of aspects 168-176, wherein ^X99 is E.

Aspect 178. 178. An IL-2Rγc ligand according to any one of aspects 168-177, wherein X ¹⁰⁰ is W.

Aspect 179. 179. An IL-2Rγc ligand according to any one of aspects 168-178, wherein X ¹⁰¹ is selected from I, L, and V.

Aspect 180. 179. An IL-2Rγc ligand according to any one of aspects 168-179, wherein X ¹⁰² is selected from D and G.

Aspect 181. 181. An IL-2Rγc ligand according to any one of aspects 168-180, wherein X ¹⁰³ is selected from S and T.

Aspect 182. X ⁹¹ is selected from D and E, X ⁹² is selected from amino acids, X ⁹³ is selected from D and F, X ⁹⁴ is S, X ⁹⁵ is selected from D and E, X ⁹⁶ is selected from amino acids, X ⁹⁷ is selected from L, M and W, X ⁹⁸ is G, X ⁹⁹ is E, X ¹⁰⁰ is W, X ¹⁰¹ is 169. An IL-2Rγc ligand according to aspect 168, wherein X ¹⁰² is selected from D and G, and X ¹⁰³ is selected from S and T.

Aspect 183. the IL-2Rγc ligand comprises the amino acid sequence of Formula (7) (SEQ ID NO:246) or Formula (7a) (SEQ ID NO:247);
^{-X114 -X115 -CX116 -X117 -X118} - (7)
^{-X111 -X112 -X113 -X114 -X115 -CX116 -X117 -X108 -X119 -X120 -X121} - (7a)
wherein X ¹¹¹ is selected from D, G, I, and Q; X ¹¹² is selected from D, I, and L; X ¹¹³ is selected from G, L, M, R, S, and Y; X ¹¹⁴ is selected from D, E, S, T, and Y; X ¹¹⁵ is selected from E, L, P, and Q; X ¹¹⁶ is selected from D, E, L, S, and T, X ¹¹⁷ is selected from F, S, and W, X ¹¹⁸ is selected from F, N, W, and Y, and X ¹¹⁹ is selected from F, I, L, and R , X ¹²⁰ is selected from A, E, L, and S, and X ¹²¹ is selected from K, N, Q, and V. IL-2Rγc ligand according to any one of aspects 74-81. .

Aspect 184. 184. An IL-2Rγc ligand according to aspect 183, wherein X ¹¹¹ is selected from D and Q.

Aspect 185. 185. An IL-2Rγc ligand according to any one of aspects 183-184, wherein X ¹¹² is selected from I and L.

Aspect 186. 186. An IL-2Rγc ligand according to any one of aspects 183-185, wherein X ¹¹³ is selected from G, L, M, R, S, and Y.

Aspect 187. 187. An IL-2Rγc ligand according to any one of aspects 183-186, wherein X ¹¹⁴ is L.

Aspect 188. 188. An IL-2Rγc ligand according to any one of aspects 183-187, wherein X ¹⁵ is selected from E and Q.

Aspect 189. 189. An IL-2Rγc ligand according to any one of aspects 183-188, wherein X ¹¹⁶ is selected from D and E.

Aspect 190. 189. An IL-2Rγc ligand according to any one of aspects 183-189, wherein X ¹¹⁷ is selected from F and W.

Aspect 191. 191. An IL-2Rγc ligand according to any one of aspects 183-190, wherein X ¹¹⁸ is selected from F, W, and Y.

Aspect 192. 192. An IL-2Rγc ligand according to any one of aspects 183-191, wherein X ¹¹⁹ is selected from F, I, and L.

Aspect 193. 193. An IL-2Rγc ligand according to any one of aspects 183-192, wherein X ¹²⁰ is S.

Aspect 194. 194. An IL-2Rγc ligand according to any one of aspects 183-193, wherein X ¹²¹ is selected from N and Q.

Aspect 195. X ¹¹¹ is selected from D and Q, X ¹¹² is selected from I and L, X ¹¹³ is selected from G, L, M, R, S, and Y, X ¹¹⁴ is L, X ¹¹⁵ is selected from E and Q, X ¹¹⁶ is selected from D and E, X ¹¹⁷ is selected from F and W, X ¹¹⁸ is selected from F, W and Y, X ¹¹⁹ is 184. An IL-2Rγc ligand according to aspect 183, wherein X ¹²⁰ is S and X ¹²¹ is selected from N and Q.

Aspect 196. 82. An IL-2Rγc ligand according to any one of aspects 74-81, wherein the IL-2Rγc ligand comprises an amino acid sequence selected from any one of SEQ ID NOs:248-254.

Aspect 197. The IL-2Rγc ligand comprises an amino acid sequence selected from any one of SEQ ID NOs:248-254, wherein the amino acid sequence is on the N-terminus, the C-terminus, or both the N-terminus and the C-terminus 197. An IL-2Rγc ligand according to aspect 196, which ends with the amino acids -GG of.

Aspect 198. The IL-2Rβ ligand comprises an amino acid sequence selected from any one of SEQ ID NOs:248-254, each amino acid independently having the following conservative substitutions: alanine (A), glycine ( G), amino acids with small hydrophobic side chains including proline (P), serine (S), and threonine (T); amino acids with hydroxyl-containing side chains including serine (S) or threonine (T); aspartic acid (D) and amino acids with acidic side chains, including glutamic acid (E); polar, including histidine (H), asparagine (N), glutamine (Q), serine (S), threonine (T), and tyrosine (Y). amino acids with neutral side chains; amino acids with basic side chains including arginine (R), lysine (K), or histidine (H); and isoleucine (I), leucine (L), methionine (M), valine 198. according to any one of aspects 196-197, comprising one or more of amino acids with large hydrophobic side chains, including (V), phenylphenyl (F), tyrosine (Y), or tyrosine (W) IL-2Rβ ligand of.

Aspect 199. the IL-2Rγc ligand comprises the amino acid sequence of Formula (7) (SEQ ID NO:246) or Formula (7a) (SEQ ID NO:247);
^{-X114 -X115 -CX116 -X117 -X118} - (7)
^{-X111 -X112 -X113 -X114 -X115 -CX116 -X117 -X108 -X119 -X120 -X121} - (7a)
wherein X ¹¹¹ is selected from amino acids, X ¹¹² is selected from amino acids containing large hydrophobic or acidic side chains, X ¹¹³ is selected from amino acids, and X ¹¹⁴ is acidic side chains or hydroxyl X ¹¹⁵ is selected from amino acids, X ¹¹⁶ is selected from amino acids, X ¹¹⁷ is selected from amino acids containing large hydrophobic side chains, and X ¹¹⁸ is selected from amino acids containing large hydrophobic side chains. X ¹¹⁹ is selected from amino acids containing large hydrophobic side chains, X ¹²⁰ is selected from amino acids, and X ¹²¹ is selected from amino acids. An IL-2Rγc ligand according to any one.

Aspect 200. X ¹¹¹ is selected from amino acids, X ¹¹² is selected from amino acids containing large hydrophobic or acidic side chains, X ¹¹³ is selected from amino acids, and X ¹¹⁴ is acidic or hydroxyl-containing side chains. X ¹¹⁵ is selected from amino acids containing a large hydrophobic side chain, X ¹¹⁶ is selected from amino acids containing an acidic side chain, and X ¹¹⁷ is selected from amino acids containing a large hydrophobic side chain X ¹¹⁸ is selected from amino acids containing large hydrophobic side chains, X ¹¹⁹ is selected from amino acids containing large hydrophobic side chains, X ¹²⁰ is selected from amino acids, X ¹²¹ is medium polar 200. An IL-2Rγc ligand according to aspect 199, wherein the IL-2Rγc ligand is selected from amino acids comprising a positive side chain.

Aspect 201. X ¹¹¹ is selected from amino acids, X ¹¹² is selected from D, E, F, I, L, M, V, Y, and W, X ¹¹³ is selected from amino acids, X ¹¹⁴ is D, E, S, and T; X ¹¹⁵ is selected from F, I, L, M, ^V , Y, and W; X ¹¹⁶ is selected from D and E; , L, M, V, Y, and W, X ¹¹⁸ is selected from F, I, L, M, V, Y, and W, and X ¹¹⁹ is selected from F, I, L, M, V , Y, and W, X ¹²⁰ is selected from amino acids, and X ¹²¹ is selected from H, N, Q, S, T, and Y.

Aspect 202. 202. An IL-2Rγc ligand according to aspect 201, wherein X ¹¹¹ is selected from amino acids.

Aspect 203. 203. An IL-2Rγc ligand according to any one of aspects 201-202, wherein X ¹¹² is selected from I and L.

Aspect 204. 204. An IL-2Rγc ligand according to any one of aspects 201-203, wherein X ¹¹³ is selected from amino acids.

Aspect 205. 205. An IL-2Rγc ligand according to any one of aspects 201-204, wherein X ¹¹⁴ is selected from D, E, and S.

Aspect 206. 206. An IL-2Rγc ligand according to any one of aspects 201-205, wherein X ¹¹⁵ is L.

Aspect 207. 207. An IL-2Rγc ligand according to any one of aspects 201-206, wherein X ¹¹⁶ is selected from D and E.

Aspect 208. 208. An IL-2Rγc ligand according to any one of aspects 201-207, wherein X ¹¹⁷ is selected from F and W.

Aspect 209. 209. An IL-2Rγc ligand according to any one of aspects 201-208, wherein X ¹¹⁸ is selected from F, W and Y.

Aspect 210. 209. An IL-2Rγc ligand according to any one of aspects 201-209, wherein X ¹¹⁹ is selected from F, I, and L.

Aspect 211. 211. An IL-2Rγc ligand according to any one of aspects 201-210, wherein X ¹²⁰ is selected from amino acids.

Aspect 212. 212. An IL-2Rγc ligand according to any one of aspects 201-211, wherein X ¹²¹ is selected from Q and N.

Aspect 213. X ¹¹¹ is selected from amino acids, X ¹¹² is selected from I and L, X ¹¹³ is selected from amino acids, X ¹¹⁴ is selected from D, E and S, X ¹¹⁵ is selected from L , X ¹¹⁶ is selected from D and E, X ¹¹⁷ is selected from F and W, X ¹¹⁸ is selected from F, W, and Y, X ¹¹⁹ is selected from F, I, and L 202, wherein X ¹²⁰ is selected from amino acids and X ¹²¹ is selected from Q and N.

Aspect 214. 82. An IL-2Rγc ligand according to any one of aspects 74-81, wherein the IL-2Rγc ligand comprises an amino acid sequence selected from any one of SEQ ID NOs:255-267.

Aspect 215. wherein the IL-2Rγc ligand comprises an amino acid sequence selected from any one of SEQ ID NOs:255-267, wherein the amino acid sequence is on the N-terminus, the C-terminus, or both the N-terminus and the C-terminus 215. An IL-2Rγc ligand according to aspect 214, which ends with amino acids -GG of.

Aspect 216. The IL-2Rβ ligand comprises an amino acid sequence selected from any one of SEQ ID NOs:255-267, each amino acid independently having the following conservative substitutions: alanine (A), glycine ( G), amino acids with small hydrophobic side chains including proline (P), serine (S), and threonine (T); amino acids with hydroxyl-containing side chains including serine (S) or threonine (T); aspartic acid (D) and amino acids with acidic side chains, including glutamic acid (E); polar, including histidine (H), asparagine (N), glutamine (Q), serine (S), threonine (T), and tyrosine (Y). amino acids with neutral side chains; amino acids with basic side chains including arginine (R), lysine (K), or histidine (H); and isoleucine (I), leucine (L), methionine (M), valine 216. According to any one of aspects 214-215, comprising one or more of amino acids with large hydrophobic side chains, including (V), phenylphenyl (F), tyrosine (Y), or tyrosine (W). IL-2Rβ ligand of.

Aspect 217. A pH-selective IL-2Rβ ligand comprising an amino acid sequence selected from ligands having SEQ ID NOs:400-577.

Aspect 218. 218. The pH-selective IL-2Rβ ligand according to aspect 217, wherein the pH-selective IL-2Rβ ligand exhibits greater than 15% reduction in binding to IL-2Rβ receptor at pH 7.5 compared to pH 6.0.

Aspect 219. the ligand is SEQ ID NO: 400, 402-405, 407, 409, 410, 411, 413, 415, 416, 418, 419, 420, 421, 423, 425-432, 436, 438-440, 442-446, 448 , 450, 452, 453-456, 459-461, 463-468, 470, 471, 473-477, 479, 481-486, 489, 491, 493-496, 498-507, 510-519, 521-524 , 526-531, 534-537, 543, 545-548, 550, 551, 558-564, 566, 568-573, and 575, wherein pH-selective binding is performed A pH-selective IL-2Rβ ligand according to aspect 518, determined as described in Examples 9-12.

Aspect 220. 218. The pH-selective IL-2Rβ ligand according to aspect 217, wherein the pH-selective IL-2Rβ ligand exhibits greater than 50% reduction in binding to IL-2Rβ receptor at pH 7.5 compared to pH 6.0.

Aspect 221. the ligand is SEQ ID NO: 400, 404-405, 407, 409, 410, 413, 415, 420, 426, 431, 432, 438-440, 442, 444-446, 450, 452, 453, 455, 459, 464 ~467, 473-475, 479, 480, 482-484, 486, 489, 493, 496, 498, 502, 504, 510-514, 516-519, 521-523, 527-530, 537, 543, 545 Embodiment 520 comprising an amino acid sequence selected from ligands having -547, 550, 558-564, 571-573, and 575, wherein the pH-selective binding is determined as described in Examples 9-12. A pH-selective IL-2Rβ ligand as described in .

Aspect 222. The pH-selective IL-2Rβ ligand comprises an amino acid sequence selected from any one of SEQ ID NOs:400-577, wherein the amino acid sequence is on the N-terminus, the C-terminus, or both the N-terminus and the C-terminus. pH-selective IL-2Rβ ligand according to any one of aspects 217-221, ending with amino acids -GG above

Aspect 223. The pH-selective IL-2Rβ ligand is an amino acid sequence selected from any one of SEQ ID NOs:400-577, each amino acid independently having the following conservative substitutions: alanine (A); amino acids with small hydrophobic side chains including glycine (G), proline (P), serine (S), and threonine (T); amino acids with hydroxyl-containing side chains including serine (S) or threonine (T); Amino acids with acidic side chains including aspartic acid (D) and glutamic acid (E); histidine (H), asparagine (N), glutamine (Q), serine (S), threonine (T), and tyrosine (Y). amino acids with polar neutral side chains, including; amino acids with basic side chains, including arginine (R), lysine (K), or histidine (H); and isoleucine (I), leucine (L), methionine (M). , valine (V), phenylphenyl (F), tyrosine (Y), or tyrosine (W). A pH-selective IL-2Rβ ligand as described in .

Aspect 224. The pH-selective IL-2Rβ ligand according to any one of aspects 217-223, wherein the pH-selective IL-2Rβ ligand comprises 5-30 amino acids.

Aspect 225. pH-selective according to any one of aspects 217-224, wherein the pH-selective IL-2Rβ ligand exhibits a binding affinity (IC ₅₀ ) for human IL-2Rβ subunits from 1 pM to 100 μM at pH 6.0 IL-2Rβ ligand.

Aspect 226. pH according to any one of aspects 217-224, wherein the pH-selective IL-2Rβ ligand exhibits a binding affinity (IC ₅₀ ) for human IL-2Rβ subunits of 0.1 μM to 50 μM at pH 6.0 Selective IL-2Rβ ligand.

Aspect 227. pH-selective IL according to any one of aspects 217-224, wherein the pH-selective IL-2Rβ ligand exhibits a binding affinity (IC ₅₀ ) for human IL-2Rβ subunits of less than 100 μM at pH 6.0. -2Rβ ligand.

Aspect 228. pH-selective according to any one of aspects 217-224, wherein the pH-selective IL-2Rβ ligand exhibits a binding affinity (IC ₅₀ ) for mammalian IL-2Rβ subunits of less than 100 μM at pH 6.0 IL-2Rβ ligand.

Aspect 229. in any one of aspects 217-228, wherein the pH-selective IL-2Rβ ligand exhibits a binding affinity (IC ₅₀ ) of less than 100 μM for each of the human IL-2Rβ subunit and the human IL-2Rγc subunit A pH-selective IL-2Rβ ligand as described.

Aspect 230. pH-selective IL- according to any one of aspects 217-229, wherein the pH-selective IL-2Rβ ligand exhibits a binding affinity (IC ₅₀ ) for the human IL-2Rα (CD25) subunit of greater than 100 μM 2Rβ ligand.

Aspect 231. The pH-selective IL-2Rβ ligand exhibits a binding affinity (IC ₅₀ ) for the human IL-2Rβ subunit that is at least 10-fold greater than the binding affinity (IC ₅₀ ) of the IL-2Rβ ligand for the human IL-2Rα subunit. A pH-selective IL-2Rβ ligand according to any one of aspects 217-230, wherein:

Aspect 232. the IL-2Rβ ligand of any one of aspects 1-73 and 217-231, the IL-2Rγc ligand of any one of aspects 74-216, or the IL-2Rβ ligand of any one of aspects 1-73, and aspect 74 A compound comprising any one IL-2Rγc ligand of -216.

Aspect 233. 233. A compound according to aspect 232, wherein the compound is selected from peptides, conjugates, fusion proteins, and single peptide chains.

Aspect 234. 234. A compound according to aspect 233, wherein the compound is a peptide.

Aspect 235. 235. A compound according to aspect 234, wherein the peptide has a molecular weight in the range of 500 Daltons to 15,000 Daltons.

Aspect 236. A compound according to any one of aspects 234-235, wherein the peptide comprises 5-5,000 amino acids.

Aspect 237. A compound according to aspect 233, wherein the compound comprises a conjugate.

Aspect 238. 238. A compound according to aspect 237, wherein the conjugate comprises at least one IL-2Rβ ligand.

Aspect 239. 238. A compound according to aspect 237, wherein the conjugate comprises at least two IL-2Rβ ligands and at least one linker attached to each of said at least two IL-2Rβ ligands.

Aspect 240. The compound of any one of aspects 237-239, wherein the conjugate comprises at least one IL-2Rγc ligand.

Aspect 241. 238. A compound according to aspect 237, wherein the conjugate comprises at least two IL-2Rγc ligands and at least one linker attached to each of said at least two IL-2Rγc ligands.

Aspect 242. According to aspect 237, wherein the conjugate comprises at least one IL-2Rβ ligand, at least one IL-2Rγc ligand, and at least one linker attached to the at least one IL-2Rβ ligand and the at least one IL-2Rγc ligand compound.

Aspect 243. A compound according to any one of aspects 237-228, wherein the conjugate comprises at least one moiety, amino acid, or polypeptide configured to modify the properties of the conjugate.

Aspect 244. 244. A compound according to aspect 243, wherein the properties are selected from water solubility, polarity, lipophilicity, pharmacokinetic profile, targeting, bioavailability, pH dependent binding, and cage (reversible loss of potency).

Aspect 245. 245. A compound according to any one of aspects 243-244, wherein at least one moiety is cleavable in vivo.

Aspect 246. 246. A compound according to any one of aspects 243-245, wherein at least one moiety comprises an irreversibly cleavable pro-moiety.

Aspect 247. 247. A compound according to aspect 246, wherein the pro-ingredient is configured to be releasable in a target-specific environment.

Aspect 248. 248. A compound according to aspect 247, wherein the target-specific environment comprises an enzyme, pH, or a combination thereof.

Aspect 249. 249. A compound according to aspect 248, wherein the moiety comprises a polymer, peptide, antibody, or combination of any of the foregoing.

Aspect 250. 244. A compound according to aspect 243, comprising a pharmacokinetic moiety.

Aspect 251. 251. A compound according to aspect 250, wherein the pharmacokinetic moiety comprises polyethylene glycol.

Aspect 252. 244. A compound according to aspect 243, comprising a tumor-targeting moiety.

Aspect 253. 253. A compound according to aspect 252, wherein the tumor-targeting moiety comprises a tumor-specific antibody, tumor-specific antibody fragment, tumor-specific protein, tumor-specific peptide, or a combination of any of the foregoing.

Aspect 254. 233. A compound according to aspect 232, comprising a linker covalently attached to an IL-2Rβ ligand, an IL-2Rγc ligand, or a combination thereof.

Aspect 255. 255. A compound according to aspect 254, wherein the linker is a peptide having 5-50 amino acids.

Aspect 256. 255. A compound according to aspect 254, wherein the linker comprises polyethylene glycol.

Aspect 257. The compound comprises a heterodimer, the heterodimer comprising an IL-2Rβ ligand, an IL-2Rγc ligand, and a linker, each of the IL-2Rβ ligand and the IL-2Rγc ligand having an amino-terminus (N-terminus), the carboxy-terminus (C-terminus), and amino acid side chains, wherein the IL-2Rβ ligand is attached to the linker via the amino-terminus (N-terminus), the carboxy-terminus (C-terminus), the amino acid side chains, or any combination of the foregoing; 233. A compound according to aspect 232, wherein the IL-2Rγc ligand is attached to the linker via the amino-terminus (N-terminus), the carboxy-terminus (C-terminus), the amino acid side chain, or a combination of any of the foregoing.

Aspect 258. The conjugate comprises a heterodimer, the heterodimer comprising an IL-2Rβ ligand, an IL-2Rγc ligand, and a linker, wherein each of the IL-2Rβ ligand and the IL-2Rγc ligand are amino-terminal (N-terminal) and the carboxy terminus (C-terminus), wherein each of the IL-2Rβ ligand and IL-2Rγc ligand is covalently attached to a linker.

Aspect 259. 259. A compound according to aspect 258, wherein each of the IL-2Rβ ligand and IL-2Rγc ligand is covalently attached to the linker via their respective C-termini.

Aspect 260. 259. A compound according to aspect 258, wherein the N-terminus of the IL-2Rβ ligand is covalently linked to the linker and the C-terminus of the IL-2Rγc ligand is covalently linked to the linker.

Aspect 261. 259. A compound according to aspect 258, wherein the C-terminus of the IL-2Rβ ligand is covalently linked to the linker and the N-terminus of the IL-2Rγc ligand is covalently linked to the linker.

Aspect 262. 259. A compound according to aspect 258, wherein the IL-2Rβ ligand is covalently attached to the linker via an amino acid side chain and the IL-2Rγc ligand is covalently attached to the linker via an amino acid side chain.

Aspect 263. The IL-2Rβ ligand is covalently attached to the linker via the amino acid side chain, via the C-terminus, or via the N-terminus, and the IL-2Rγc ligand is covalently attached via the amino acid side chain, via the C-terminus, to the linker. or a compound according to aspect 258, covalently attached via the N-terminus to a linker.

Aspect 264. 259. A compound according to aspect 258, wherein the heterodimer is configured to activate the IL-2 receptor.

Aspect 265. A compound according to aspect 258, wherein the linker is configured such that the heterodimer activates the IL-2 receptor.

Aspect 266. 266. Aspect 258-265, wherein primary human peripheral blood mononuclear cells (PBMC) expressing the human IL-2Rβγc subunit phosphorylate transcription 5 (STAT5) when incubated with the heterodimer Compound as described.

Aspect 267. 267. Any of aspects 258-266, wherein primary human peripheral blood mononuclear cells (PBMC) expressing human IL-2Rα [CD25] subunits do not phosphorylate transcript 5 (STAT5) when incubated with heterodimers A compound according to item 1.

Aspect 268. 268. A compound according to any one of aspects 258-267, wherein the heterodimer comprises a conformation configured to activate the human IL-2Rβγc signaling pathway.

Aspect 269. 233. A compound according to aspect 232, wherein the compound comprises a single peptide chain.

Aspect 270. A compound according to aspect 269, wherein the single peptide chain comprises at least one IL-2Rβ ligand.

Aspect 271. 270. A compound according to aspect 269, wherein the single peptide chain comprises at least two IL-2Rβ ligands and at least one linker attached to the at least two IL-2Rβ ligands.

Aspect 272. 270. A compound according to aspect 269, wherein the single peptide chain comprises at least one IL-2Rγc ligand.

Aspect 273. 269. A compound according to aspect 269, wherein the single peptide chain comprises at least two IL-2Rγc ligands and at least one linker attached to the at least two IL-2Rγc ligands.

Aspect 274. Embodiment 269, wherein the single-chain peptide comprises at least one IL-2Rβ ligand, at least one IL-2Rγc ligand, and at least one linker attached to the at least one IL-2Rβ ligand and the at least one IL-2Rγc ligand The compound described in .

Aspect 275. 233. A compound according to aspect 232, wherein the compound is a fusion protein.

Aspect 276. 276. A compound according to aspect 275, wherein the fusion protein comprises an IL-2Rβ ligand, an IL-2Rγc ligand, and a peptide linker domain, wherein the peptide linker domain binds the IL-2Rβ ligand and the IL-2Rγc ligand.

Aspect 277. Each domain has an amino-terminus (N-terminus) and a carboxy-terminus (C-terminus), and the fusion protein is such that the C-terminus and the human IL-2 variant protein domain are fused through a peptide bond to the N-terminus of a peptide linker domain. 276. A compound according to aspect 275, wherein the N-terminus of the peptide linker domain is configured such that the N-terminus of the IgG Fc protein domain is fused through a peptide bond to the C-terminus of the peptide linker domain.

Aspect 278. A nucleic acid encoding the fusion protein of aspect 275.

Aspect 279. 233. A compound according to aspect 232, wherein the compound comprises a label.

Aspect 280. 280. A compound according to aspect 279, wherein the label is selected from a radioisotope, a fluorophore, or a combination thereof.

Aspect 281. 233. A compound according to aspect 232, wherein the compound comprises a cage to protect peripheral tissue against the toxicity of IL-2R activation.

Aspect 282. A compound according to aspect 232, wherein the compound comprises a prodrug.

Aspect 283. 283. A compound according to aspect 282, wherein the compound comprises a moiety configured to maintain a circulating reservoir of prodrug.

Aspect 284. 233. A compound according to aspect 232, wherein the compound comprises a moiety configured to target IL-2R-directed immune stimulation of effector immune cells in tumors.

Aspect 285. an IL-2Rβ ligand according to any one of aspects 1-73 and 217-231, an IL-2Rγc ligand according to any one of aspects 74-216, an IL-2Rγc ligand according to any one of aspects 232-284 A pharmaceutical composition comprising a compound, or a combination of any of the foregoing.

Aspect 286. 271. A method of treating cancer in a patient, comprising administering a therapeutically effective amount of a pharmaceutical composition according to aspect 270 to a patient in need of such treatment.

Aspect 287. A method of treating cancer in a patient comprising administering to a patient in need of such treatment a therapeutically effective amount of an IL-2Rβ ligand of any one of aspects 1-73 and 217-231, aspects 74-216 or a compound according to any one of aspects 232-284, or a combination of any of the foregoing.

Aspect 288. 288. The method of aspect 287, wherein the cancer comprises a solid tumor.

Aspect 289. 286. A method of treating an autoimmune disease in a patient, comprising administering a therapeutically effective amount of a pharmaceutical composition according to aspect 285 to a patient in need of such treatment.

Aspect 290. A method of treating an autoimmune disease in a patient comprising administering to the patient in need of such treatment a therapeutically effective amount of an IL-2Rβ ligand of any one of aspects 1-73 and 217-231, aspects 74- 216, a compound according to any one of aspects 232-284, or a combination of any of the foregoing.

Aspect 291. A method of screening a compound for IL-2 receptor activity, wherein a cell is treated with an IL-2Rβ ligand of any one of aspects 1-73 and 217-231, an IL-2Rγc ligand of any one of aspects 74-216 Aspects 232-284, or a combination of any of the foregoing, wherein the cell expresses the IL-2 receptor; A method comprising contacting with a test compound and determining the activity of the test compound.

Aspect 292. A method of activating a human IL-2Rβ subunit, a human IL-2Rγc subunit, or both a human IL-2Rβ subunit and a human IL-2Rγc subunit, wherein the IL-2 receptor is expressed in vivo the cell with an IL-2Rβ ligand of any one of aspects 1-73 and 217-231, an IL-2Rγc ligand of any one of aspects 74-216, a compound according to any one of aspects 232-284, or A method comprising contacting with a combination of any of the foregoing.

Aspect 293. A method of activating human IL-2Rβ subunit and human IL-2Rγc subunit in a patient comprising an effective amount of an IL-2Rβ ligand of any one of aspects 1-73 and 217-231, aspects 74-216 or a compound according to any one of aspects 232-284, or a combination of any of the foregoing, to the patient.

Aspect 294. A method of treating a disease in a patient, wherein the IL-2 receptor signaling pathway is implicated in disease etiology and in need of such treatment, administering to a patient in need of such treatment a therapeutically effective amount of embodiments 1-73 and 217- administering an IL-2Rβ ligand of any one of 231, an IL-2Rγc ligand of any one of aspects 74-216, a compound according to any one of aspects 232-284, or a combination of any of the foregoing A method comprising:

Aspect 295. A method of treating a disease in a patient, wherein activation of the IL-2 receptor is effective in treating the disease and administering to a patient in need of such treatment a therapeutically effective amount of aspects 1-73 and 217 231, an IL-2Rγc ligand of any one of aspects 74-216, a compound according to any one of aspects 232-284, or a combination of any of the foregoing A method comprising administering.

Embodiment 1A. An IL-2Rβ ligand that exhibits a binding affinity (IC ₅₀ ) for human IL-2Rβ subunits of less than 100 μM.

Embodiment 2A. An IL-2Rβ ligand according to aspect 1A, wherein the IL-2Rβ ligand comprises 5-30 amino acids.

Embodiment 3A. An IL-2Rβ ligand according to any one of aspects 1A-2A, wherein the IL-2Rβ ligand exhibits a binding affinity (IC ₅₀ ) for human IL-2Rβ subunits between 1 pM and 100 μM.

Embodiment 4A. An IL-2Rβ ligand according to any one of aspects 1A-2A, wherein the IL-2Rβ ligand exhibits a binding affinity (IC ₅₀ ) for human IL-2Rβ subunits between 0.1 μM and 50 μM.

Embodiment 5A. An IL-2Rβ ligand according to any one of aspects 1A-2A, wherein the IL-2Rβ ligand exhibits a binding affinity (IC ₅₀ ) for human IL-2Rβ subunits of less than 100 μM.

Embodiment 6A. An IL-2Rβ ligand according to any one of aspects 1A-2A, wherein the IL-2Rβ ligand exhibits a binding affinity (IC ₅₀ ) for mammalian IL-2Rβ subunits of less than 100 μM.

Embodiment 7A. The IL of any one of aspects 1A-6A, wherein the IL-2Rβ ligand exhibits a binding affinity (IC ₅₀ ) of less than 100 μM for each of the human IL-2Rβ subunit and the human IL-2Rγc subunit. -2Rβ ligand.

Embodiment 8A. An IL-2Rβ ligand according to any one of aspects 1A-7A, wherein the IL-2Rβ ligand exhibits a binding affinity (IC ₅₀ ) for the human IL-2Rα (CD25) subunit of greater than 100 μM.

Embodiment 9A. An embodiment wherein the IL-2Rβ ligand exhibits a binding affinity (IC ₅₀ ) for the human IL-2Rβ subunit that is at least 10-fold greater than the binding affinity (IC ₅₀ ) of the IL-2Rβ ligand for the human IL-2Rα subunit. IL-2Rβ ligand according to any one of 1A-7A.

Embodiment 10A. the IL-2Rβ ligand comprises the amino acid sequence of formula (1) (SEQ ID NO:1), the amino acid sequence of formula (1a) (SEQ ID NO:2), or the amino acid sequence of formula (1b) (SEQ ID NO:3);
^{-X3 -X4 -X5 -X6 -X7 -X8 -X9 -X10} - (1)
^-X2 -CX3 ^{-X4 -X5 -X6 -X7 -X8 -X9 -X10 -CX11} - ( ^1a )
^{-X1 -X2} -CX3 ^{-X4 -X5 -X6 -X7 -X8 -X9 -X10 -CX11 -X12 -} (1b)
wherein X ¹ is selected from A, D, E, F, G, I, K, L, M, N, P, Q, S, T, V, W, and ^Y ; , C, D, E, F, G, H, K, L, N, P, R, S, T, W, and Y, and X ³ are selected from A, D, E, F, G, H , M, N, Q, R, S, T, W, and Y, and X ⁴ is selected from A, D, E, F, G, I, K, L, M, N, Q, R, S , T, V, and Y, X ⁵ is selected from A, G, I, Q, S, T, V, and W, X ⁶ is selected from A, D, E, G, H, K , L, M, N, P, Q, R, S, T, and V; X ⁷ is selected from F, I, K, L, Q, and V; X ⁸ is selected from D, F , G, H, M, N, W, and Y, X ⁹ is selected from A, D, E, M, P, Q, S, T, V, and W, and X ¹⁰ is selected from D , F, I, L, M, S, T, V, and Y, and X ¹¹ is selected from D, E, F, H, I, L, M, Q, S, T, V, W, and An IL-2Rβ ligand according to any one of aspects 1A-9A, wherein X ¹² is selected from F, I, L, M, N, S, V, W, and Y.

Embodiment 11A. An IL-2Rβ ligand according to aspect 10A, wherein X ¹ is selected from F, I, L, M, and V.

Embodiment 12A. IL-2Rβ according to any one of aspects 10A-11A, wherein ^X2 is selected from D, E, F, G, H, L, N, P, R, S, T, W, and Y ligand.

Embodiment 13A. An IL-2Rβ ligand according to any one of aspects 10A-12A, wherein ^X5 is A.

Embodiment 14A. An IL-2Rβ ligand according to any one of aspects 10A-13A, wherein ^X6 is selected from D, E, and Q.

Embodiment 15A. An IL-2Rβ ligand according to any one of aspects 10A-14A, wherein ^X7 is selected from F, I, L, and V.

Embodiment 16A. An IL-2Rβ ligand according to any one of aspects 10A-15A, wherein X ⁸ is G.

Embodiment 17A. An IL-2Rβ ligand according to any one of aspects 10A-16A, wherein X ⁹ is selected from D, E, and Q.

Embodiment 18A. An IL-2Rβ ligand according to any one of aspects 10A-17A, wherein X ¹⁰ is selected from F, I, L, M, V, and Y.

Embodiment 19A. An IL-2Rβ ligand according to any one of aspects 10A-18A, wherein X ¹¹ is selected from D and E.

Embodiment 20A. An IL-2Rβ ligand according to any one of aspects 10A-19A, wherein ^X12 is selected from F, I, L, M, and V.

Embodiment 21A. X ¹ is selected from F, I, L, M, and V and X ² is selected from D, E, F, G, H, L, N, P, R, S, T, W, and Y and X ³ is selected from A, D, E, F, G, H, M, N, Q, R, S, T, W, and Y, and X ⁴ is selected from A, D, E, F, is selected from G, I, K, L, M, N, Q, R, S, T, V, and Y; X ⁵ is A; X ⁶ is selected from D, E, and Q; X ⁷ is selected from F, I, L, and V, X ⁸ is G, X ⁹ is selected from D, E, and Q, X ¹⁰ is F, I, L, M, An IL-2Rβ ligand according to aspect 10A, wherein X ¹¹ is selected from V and Y, X 11 is selected from D and E, and X ¹² is selected from F, I, L, M and V.

Embodiment 22A. the IL-2Rβ ligand comprises the amino acid sequence of formula (1) (SEQ ID NO:1), the amino acid sequence of formula (1a) (SEQ ID NO:2), or the amino acid sequence of formula (1b) (SEQ ID NO:3);
^{-X3 -X4 -X5 -X6 -X7 -X8 -X9 -X10} - (1)
^-X2 -CX3 ^{-X4 -X5 -X6 -X7 -X8 -X9 -X10 -CX11} - ( ^1a )
^{-X1 -X2} -CX3 ^{-X4 -X5 -X6 -X7 -X8 -X9 -X10 -CX11 -X12 -} (1b)
wherein X ¹ is selected from amino acids, X ² is selected from amino acids, X ³ is selected from amino acids, X ⁴ is selected from amino acids, and X ⁵ contains a small hydrophobic side chain is selected from amino acids, X ⁶ is selected from amino acids, X ⁷ is selected from amino acids containing large hydrophobic side chains, X ⁸ is selected from amino acids containing small hydrophobic side chains, X ⁹ is selected from selected from amino acids containing polar neutral or acidic side chains, X ¹⁰ is selected from amino acids containing large hydrophobic side chains, X ¹¹ is selected from amino acids, and X ¹² contains large hydrophobic side chains An IL-2Rβ ligand according to any one of aspects 1A-9A, selected from amino acids.

Embodiment 23A. X ¹ is selected from amino acids containing large hydrophobic side chains, X ² is selected from amino acids, X ³ is selected from amino acids, X ⁴ is selected from amino acids, X ⁵ is selected from small hydrophobic is selected from amino acids containing side chains, X ⁶ is selected from amino acids containing polar neutral or acidic side chains, X ⁷ is selected from amino acids containing large hydrophobic side chains, and X ⁸ is selected from amino acids containing small hydrophobic side chains; selected from amino acids containing side chains, X ⁹ is selected from amino acids containing polar neutral or acidic side chains, X ¹⁰ is selected from amino acids containing large hydrophobic side chains, X ¹¹ is polar neutral or an IL-2Rβ ligand according to aspect 22A, selected from amino acids containing acidic side chains and X ¹² is selected from amino acids containing large hydrophobic side chains.

Embodiment 24A. X ¹ is selected from I, L, M, V, F, W, and Y, X ² is selected from amino acids, X ³ is selected from amino acids, X ⁴ is selected from amino acids, X ⁵ is selected from A, G, P, S, and T; X ⁶ is selected from H, N, Q, S, T, Y, D, and E; X ⁷ is selected from I, L, M , V, F, W, and Y, X ⁸ is selected from A, G, P, S, and T, and X ⁹ is H, N, Q, S, T, Y, D, and E, X ¹⁰ is selected from I, L, M, V, F, W, and Y, and X ¹¹ is selected from H, N, Q, S, T, Y, D, and E , X ¹² is selected from I, L, M, V, F, W, and Y. An IL-2Rβ ligand according to any one of aspects 22A-23A.

Embodiment 25A. X ¹ is selected from I, L, M, V, F, W, and Y, X ² is selected from amino acids, X ³ is selected from amino acids, X ⁴ is selected from amino acids, X ⁵ is A; X ⁶ is selected from H, N, Q, S, T, Y, D, and E; X ⁷ is selected from I, L, M, V, F, W, and Y is selected, X ⁸ is G, X ⁹ is selected from H, N, Q, S, T, Y, D, and E, and X ¹⁰ is I, L, M, V, F, W , and Y, X ¹¹ is selected from H, N, Q, S, T, Y, D, and E, and X ¹² is selected from I, L, M, V, F, W, and Y An IL-2Rβ ligand according to any one of aspects 22A-23A that is selected.

Embodiment 26A. An IL-2Rβ ligand according to aspect 25A, wherein X ¹ is selected from I, L, M, and V.

Embodiment 27A. An IL-2Rβ ligand according to any one of aspects 25A-26A, wherein ^X2 is selected from D and E.

Embodiment 28A. An IL-2Rβ ligand according to any one of aspects 25A-27A, wherein ^X6 is selected from Q, E, and D.

Embodiment 29A. An IL-2Rβ ligand according to any one of aspects 25A-28A, wherein ^X7 is selected from V, L, and I.

Embodiment 30A. An IL-2Rβ ligand according to any one of aspects 25A-29A, wherein X ⁹ is selected from E, D, and Q.

Embodiment 31A. An IL-2Rβ ligand according to any one of aspects 25A-30A, wherein X ¹⁰ is selected from L, V, I, and Y.

Embodiment 32A. An IL-2Rβ ligand according to any one of aspects 25A-31A, wherein X ¹¹ is selected from D and E.

Embodiment 33A. An IL-2Rβ ligand according to any one of aspects 25A-32A, wherein ^X12 is selected from L, I, and F.

Embodiment 34A. X ¹ is selected from L, I, F, and V ^; X ² is selected from D and E; X ⁶ is selected from Q, E, and D; is selected from I, X ⁹ is selected from E, D, and Q; X ¹⁰ is selected from L, V, ^{I, and Y; X 11 is} selected from D and E; An IL-2Rβ ligand according to aspect 25A, selected from L, I, and F.

Embodiment 35A. X ¹ is selected from F, I, M, and Y; X ² is selected from E, D, and R; X ³ is selected from and amino acids; X ⁴ is selected from amino acids; ⁵ is A, X ⁶ is selected from A, P, and Q, X ⁷ is selected from I and V, X ⁸ is G, and X ⁹ is selected from E and Q , X ¹⁰ is selected from I, L, and V; X ¹¹ is selected from E, D, and Q; and X ¹² is selected from I and L. .

Embodiment 36A. An IL-2Rβ ligand according to aspect 22A, wherein the IL-2Rβ ligand comprises an amino acid sequence selected from any one of SEQ ID NOs:4-163.

Embodiment 37A. the IL-2Rβ ligand comprises an amino acid sequence selected from any one of SEQ ID NOs: 4-163, wherein the amino acid sequence is on the N-terminus, the C-terminus, or both the N-terminus and the C-terminus IL-2Rβ ligand according to aspect 36A, ending with amino acids -GG-.

Embodiment 38A. The IL-2Rβ ligand comprises an amino acid sequence selected from any one of SEQ ID NOs: 4-163, each amino acid independently having the following conservative substitutions: alanine (A), glycine ( G) amino acids with small hydrophobic side chains, including proline (P), serine (S), or threonine (T); hydroxyl-containing side chains, including serine (S), threonine (T), or tyrosine (Y) amino acids with acidic side chains, including aspartic acid (D), or glutamic acid (E); histidine (H), asparagine (N), glutamine (Q), serine (S), threonine (T), or amino acids with polar neutral side chains, including tyrosine (Y); amino acids with basic side chains, including arginine (R), lysine (K), or histidine (H); isoleucine (I), leucine (L), Amino acids with large hydrophobic side chains including methionine (M), valine (V), phenylalanine (F), tyrosine (Y), or tryptophan (W); as well as phenylalanine (F), histidine (H), tryptophan (W ), or amino acids with aromatic side chains including tyrosine (Y).

Embodiment 39A. the IL-2Rβ ligand comprises the amino acid sequence of formula (2) (SEQ ID NO: 164), the amino acid sequence of formula (2a) (SEQ ID NO: 165), or the amino acid sequence of formula (2b) (SEQ ID NO: 166);
^{-X15 -X16 -X17 -X18 -X19 -X20 -X21 -X22} - (2)
^{-X14 -CX15 -X16 -X17 -X18 -X19 -X20 -X21 -X22 -CX23} - (2a)
^{-X13 -X14 -CX15 -X16 -X17 -X18 -X19 -X20 -X21 -X22 -CX23 -X24} - (2b)
wherein X ¹³ is selected from A, D, E, G, N, Q, R, and V; ^{X 14} is selected from E, F, I, L, M, and Q; , D, G, L, and N, X ¹⁶ is selected from L, P, V, and Y, X ¹⁷ is selected from F, G, and M, and X ¹⁸ is selected from A, D , N, and Q; ^{X 19} is selected from F, I, L, S, V, W, and Y; X ²⁰ is selected from D and W; X ²² is selected from A, D, Q, and S; X ²³ is selected from I, L, Q, W, and Y; X ²⁴ is selected from E, F, I, L, T , V, and W, according to any one of aspects 1A-9A.

Embodiment 40A. An IL-2Rβ ligand according to embodiment 39A, wherein ^X16 is V.

Embodiment 41A. An IL-2Rβ ligand according to any one of aspects 39A-40A, wherein ^X17 is G.

Embodiment 42A. An IL-2Rβ ligand according to any one of aspects 39A-41A, wherein ^X20 is W.

Embodiment 43A. An IL-2Rβ ligand according to any one of aspects 39A-42A, wherein ^X21 is P.

Embodiment 44A. X ¹³ is selected from E, N, and Q, X ¹⁴ is selected from I and M, X ¹⁵ is selected from D, L, and N, X ¹⁶ is V, and X ¹⁷ is , G, X ¹⁸ is selected from D and Q, X ¹⁹ is selected from V, W, and Y, X ²⁰ is W, X ²¹ is P, X ²² is An IL-2Rβ ligand according to embodiment 39A, wherein ^X23 is selected from D and S, X23 is selected from L and Q, and ^X24 is selected from I, L, and V.

Embodiment 45A. An IL-2Rβ ligand according to aspect 39A, wherein the IL-2Rβ ligand comprises an amino acid sequence selected from any one of SEQ ID NOs:167-182.

Embodiment 46A. IL-2Rβ ligand comprises an amino acid sequence selected from any one of SEQ ID NOs: 167-182, wherein the amino acid sequence is on the N-terminus, the C-terminus, or both the N-terminus and the C-terminus IL-2Rβ ligand according to aspect 45A, ending with amino acids -GG-.

Embodiment 47A. The IL-2Rβ ligand comprises an amino acid sequence selected from any one of SEQ ID NOS: 167-182, each amino acid independently having the following conservative substitutions: alanine (A), glycine ( G) amino acids with small hydrophobic side chains, including proline (P), serine (S), or threonine (T); hydroxyl-containing side chains, including serine (S), threonine (T), or tyrosine (Y) amino acids with acidic side chains, including aspartic acid (D), or glutamic acid (E); histidine (H), asparagine (N), glutamine (Q), serine (S), threonine (T), or amino acids with polar neutral side chains, including tyrosine (Y); amino acids with basic side chains, including arginine (R), lysine (K), or histidine (H); isoleucine (I), leucine (L), Amino acids with large hydrophobic side chains including methionine (M), valine (V), phenylalanine (F), tyrosine (Y), or tryptophan (W); as well as phenylalanine (F), histidine (H), tryptophan (W ), or amino acids with aromatic side chains including tyrosine (Y).

Embodiment 48A. the IL-2Rβ ligand comprises the amino acid sequence of formula (2) (SEQ ID NO: 164), the amino acid sequence of formula (2a) (SEQ ID NO: 165), or the amino acid sequence of formula (2b) (SEQ ID NO: 166);
^{-X15 -X16 -X17 -X18 -X19 -X20 -X21 -X22} - (2)
^{-X14 -CX15 -X16 -X17 -X18 -X19 -X20 -X21 -X22 -CX23} - (2a)
^{-X13 -X14 -CX15 -X16 -X17 -X18 -X19 -X20 -X21 -X22 -CX23 -X24} - (2b)
wherein X ¹³ is selected from amino acids, X ¹⁴ is selected from amino acids containing large hydrophobic side chains, X ¹⁵ is selected from amino acids, and X ¹⁶ is selected from amino acids containing large hydrophobic side chains. selected wherein X ²⁷ is selected from amino acids containing small hydrophobic side chains, X ¹⁸ is selected from amino acids, X ¹⁹ is selected from amino acids, and X ²⁰ is selected from amino acids containing large hydrophobic side chains selected wherein ^X21 is selected from amino acids containing small hydrophobic side chains, ^X22 is selected from amino acids, ^X23 is selected from amino acids, and ^X24 is selected from amino acids containing large hydrophobic side chains An IL-2Rβ ligand according to any one of aspects 1A-9A that is selected.

Embodiment 49A. X ¹³ is selected from amino acids, X ¹⁴ is selected from amino acids containing large hydrophobic side chains, X ¹⁵ is selected from amino acids, X ¹⁶ is selected from amino acids containing large hydrophobic side chains, X ¹⁷ is selected from amino acids containing small hydrophobic side chains, X ¹⁸ is selected from amino acids containing polar neutral or acidic side chains, and X ¹⁹ is selected from amino acids containing large hydrophobic or neutral side chains. is selected, X ²⁰ is selected from amino acids containing large hydrophobic side chains, X ²¹ is selected from amino acids containing small hydrophobic side chains, X ²² is selected from amino acids, and X ²³ is selected from amino acids IL-2Rβ ligand according to aspect 48A, wherein X ²⁴ is selected from amino acids containing large hydrophobic side chains.

Embodiment 50A. X ¹³ is selected from amino acids, X ¹⁴ is selected from I, L, M, V, F, W, and Y, X ¹⁵ is D, E, I, L, M, V, F, Y, and W, X ¹⁶ is selected from I, L, M, N, V, F, Y, and W; X ¹⁷ is selected from A, ^G , P, S, and T; is selected from H, N, Q, S, T, Y, D, and E, X ¹⁹ is selected from I, L, M, V, F, W, and Y, and X ²⁰ is selected from I, L, M, N, V, F, Y, and W; X ²¹ is selected from A, G, P, S, and T; X ²² is selected from amino acids; X ²³ is an amino acid and X ²⁴ is selected from I, L, M, V, F, W, and Y. IL-2Rβ ligand according to any one of aspects 48A-49A.

Embodiment 51A. An IL-2Rβ ligand according to aspect 50A, wherein ^X14 is selected from I and M.

Embodiment 52A. An IL-2Rβ ligand according to any one of aspects 50A-51A, wherein ^X16 is V.

Embodiment 53A. An IL-2Rβ ligand according to any one of aspects 50A-52A, wherein ^X17 is G.

Embodiment 54A. An IL-2Rβ ligand according to any one of aspects 50A-53A, wherein ^X18 is selected from D and Q.

Embodiment 55A. An IL-2Rβ ligand according to any one of aspects 50A-54A, wherein ^X20 is W.

Embodiment 56A. An IL-2Rβ ligand according to any one of aspects 50A-55A, wherein ^X21 is P.

Embodiment 57A. An IL-2Rβ ligand according to any one of aspects 50A-56A, wherein ^X23 is selected from F, I, L, and V.

Embodiment 58A. X ¹³ is selected from amino acids, X ¹⁴ is selected from I and M, X ¹⁵ is selected from amino acids, X ¹⁶ is V, X ¹⁷ is G, X ¹⁸ is D and Q, X ¹⁹ is selected from I, L, M, V, F, W, and Y, X ²⁰ is W, X ²¹ is P, and X ²² is from an amino acid 50A, wherein ^X23 is selected from amino acids and ^X24 is selected from F, I, L, and V. IL-2Rβ ligand according to aspect 50A.

Embodiment 59A. the IL-2Rβ ligand comprises the amino acid sequence of formula (3) (SEQ ID NO: 183) or the amino acid sequence of formula (3a) (SEQ ID NO: 184);
^{-X26 -X27 -X28 -X29 -X30 -X31 -X32 -X33 -X34 -X35} - ( ³ )
^{-X25 -CX26 -X27 -X28 -X29 -X30 -X31 -X32 -X33 -X34 -X35 -CX36} - (3a)
wherein X ²⁵ is selected from amino acids, X ²⁶ is selected from amino acids, X ²⁷ is selected from I and V, X ²⁸ is selected from G, X ²⁹ is D, E, and N, X ³⁰ is selected from F, L, and Y, X ³¹ is selected from F, I, and V, X ³² is selected from D and Q, X ³³ is from amino acids IL-2Rβ ligand according to any one of aspects 1A-9A, wherein X ³⁴ is selected from amino acids, X ³⁵ is selected from amino acids and X ³⁶ is selected from amino acids.

Embodiment 60A. X ²⁵ is selected from L, S, T, and Y ^; X ²⁶ is selected from ^H and Q; X ²⁷ is selected from I and V; D, E, and N; X ³⁰ is selected from F, L, and Y; X ³¹ is selected from F, I, and V; X ³² is selected from D and Q; ³³ is selected from D, L, and W; X ³⁴ is selected from G, L, ^{and T; X 35 is} selected from D, I, and S; An IL-2Rβ ligand according to aspect 59A.

Embodiment 61A. the IL-2Rβ ligand comprises the amino acid sequence of formula (3) (SEQ ID NO: 183) or the amino acid sequence of formula (3a) (SEQ ID NO: 184);
^{-X26 -X27 -X28 -X29 -X30} -X31 ^{-X32 -X33 -X34 -X35} - ( ³ )
^{-X25 -CX26 -X27 -X28 -X29 -X30 -X31 -X32 -X33 -X34 -X35 -CX36} - (3a)
wherein X ²⁵ is selected from amino acids, X ²⁶ is selected from amino acids, X ²⁷ is selected from amino acids containing large hydrophobic side chains, and X ²⁸ is selected from amino acids containing small hydrophobic side chains. selected wherein X ²⁹ is selected from amino acids comprising an acidic side chain or a polar neutral side chain, X ³⁰ is selected from amino acids, X ³¹ is selected from amino acids, X ³² is a polar neutral side chain or selected from amino acids comprising an acidic side chain, X ³³ is selected from amino acids, X ³⁴ is selected from amino acids, X ³⁵ is selected from amino acids, and X ³⁶ is selected from amino acids, Aspect 1A IL-2Rβ ligand according to any one of -9A.

Embodiment 62A. X ²⁵ is selected from amino acids, X ²⁶ is selected from amino acids, X ²⁷ is selected from amino acids containing large hydrophobic side chains, X ²⁸ is selected from amino acids containing small hydrophobic side chains, X ²⁹ is selected from amino acids containing acidic or polar neutral side chains, X ³⁰ is selected from amino acids containing large hydrophobic side chains, and X ³¹ is selected from amino acids containing large hydrophobic side chains. X ³² is selected from amino acids comprising a polar neutral side chain or an acidic side chain, X ³³ is selected from amino acids, X ³⁴ is selected from amino acids, X ³⁵ is selected from amino acids, X An IL-2Rβ ligand according to aspect 61A, wherein ³⁶ are selected from amino acids.

Embodiment 63A. the IL-2Rβ ligand comprises the amino acid sequence of formula (3) (SEQ ID NO: 183) or the amino acid sequence of formula (3a) (SEQ ID NO: 184);
^{-X26 -X27 -X28 -X29 -X30} -X31 ^{-X32 -X33 -X34 -X35} - ( ³ )
^{-X25 -CX26 -X27 -X28 -X29 -X30 -X31 -X32 -X33 -X34 -X35 -CX36} - (3a)
wherein ^{X 25} is selected from amino acids; X ²⁶ is selected from amino acids; X ²⁷ is selected from I, L, M, V, F, Y, and W; , P, S, and T; X ²⁹ is selected from D, E, H, N, Q, S, T, and Y; X ³⁰ is selected from I, L, M, V, F, Y , and W, X ³¹ is selected from I, L, M, V, F, Y, and W, and X ³² is selected from D, E, H, N, Q, T, and Y , X ³³ is selected from amino acids, X ³⁴ is selected from amino acids, X ³⁵ is selected from amino acids, and X ³⁶ is selected from amino acids. IL-2Rβ ligand.

Embodiment 64A. An IL-2Rβ ligand according to aspect 63A, wherein ^X27 is selected from V and I.

Embodiment 65A. An IL-2Rβ ligand according to any one of aspects 63A-64A, wherein ^X28 is G.

Embodiment 66A. An IL-2Rβ ligand according to any one of aspects 63A-65A, wherein ^X29 is selected from D and E.

Embodiment 67A. An IL-2Rβ ligand according to any one of aspects 63A-66A, wherein ^X30 is selected from V, L, F, and Y.

Embodiment 68A. An IL-2Rβ ligand according to any one of aspects 63A-67A, wherein ^X31 is selected from I, V, and F.

Embodiment 69A. An IL-2Rβ ligand according to any one of aspects 63A-68A, wherein ^X32 is selected from Q and D.

Embodiment 70A. X ²⁵ is selected from amino acids, X ²⁶ is selected from amino acids, X ²⁷ is selected from V and I, X ²⁸ is selected from G, X ²⁹ is selected from D and E, X ³⁰ is selected from V, L, F, and Y; X ³¹ is selected from I, V, and F; X ³² is selected from Q and D; X ³³ is selected from amino acids; IL-2Rβ ligand according to aspect 63A, wherein ³⁴ is selected from amino acids, X ³⁵ is selected from amino acids and X ³⁶ is selected from amino acids

Embodiment 71A. An IL-2Rβ ligand according to aspect 63A, wherein the IL-2Rβ ligand comprises an amino acid sequence selected from any one of SEQ ID NOs:185-193.

Embodiment 72A. IL-2Rβ ligand comprises an amino acid sequence selected from any one of SEQ ID NOs: 185-193, wherein the amino acid sequence is on the N-terminus, the C-terminus, or both the N-terminus and the C-terminus 71A, wherein the IL-2Rβ ligand according to aspect 71A ends with amino acids -GG-.

Embodiment 73A. The IL-2Rβ ligand comprises an amino acid sequence selected from any one of SEQ ID NOS: 185-193, each amino acid independently having the following conservative substitutions: alanine (A), glycine ( G) amino acids with small hydrophobic side chains, including proline (P), serine (S), or threonine (T); hydroxyl-containing side chains, including serine (S), threonine (T), or tyrosine (Y) amino acids with acidic side chains, including aspartic acid (D), or glutamic acid (E); histidine (H), asparagine (N), glutamine (Q), serine (S), threonine (T), or amino acids with polar neutral side chains, including tyrosine (Y); amino acids with basic side chains, including arginine (R), lysine (K), or histidine (H); isoleucine (I), leucine (L), Amino acids with large hydrophobic side chains including methionine (M), valine (V), phenylalanine (F), tyrosine (Y), or tryptophan (W); as well as phenylalanine (F), histidine (H), tryptophan (W ), or amino acids with aromatic side chains including tyrosine (Y).

Embodiment 74A. the IL-2Rβ ligand comprises the amino acid sequence of formula (13) (SEQ ID NO: 1028);
-X ²⁰¹ -X ²⁰² -X ²⁰³ -X ²⁰⁴ -X ²⁰⁵ -X ²⁰⁶ -X ²⁰⁷ -X ²⁰⁸ -X ²⁰⁹ -X ²¹⁰ -X ²¹¹ -X ²¹² - (13)
wherein X ²⁰¹ is selected from amino acids, X ²⁰² is selected from amino acids, X ²⁰³ is selected from amino acids containing acidic side chains, and X ²⁰⁴ is selected from amino acids containing large hydrophobic side chains. , X ²⁰⁵ is selected from amino acids containing small hydrophobic side chains, X ²⁰⁶ is selected from amino acids containing acidic side chains, X ²⁰⁷ is selected from amino acids containing large hydrophobic side chains, and X ²⁰⁸ is selected from , X ²⁰⁹ is selected from amino acids containing an acidic side chain, X ²¹⁰ is selected from amino acids, X ²¹¹ is selected from amino acids, and X ²¹² is an amino acid containing a large hydrophobic side chain. The IL-2Rβ ligand of any one of aspects 1A-9A, selected from:

Embodiment 75A. X ²⁰¹ is selected from amino acids, X ²⁰² is selected from amino acids, X ²⁰³ is selected from D and E, X ²⁰⁴ is selected from I, L, M, V, F, Y, and W , X ²⁰⁵ is selected from A, G, P, S, and T, X ²⁰⁶ is selected from D and E, and X ²⁰⁷ is selected from I, L, M, V, F, Y, and W X ²⁰⁸ is selected from amino acids, X ²⁰⁹ is selected from D and E, X ²¹⁰ is selected from amino acids, X ^{211 is} selected from amino acids, X ²¹² is selected from I, L, M, IL-2Rβ ligand according to aspect 74A, selected from V, F, Y, and W

Embodiment 76A. X ²⁰¹ is selected from C, F, L, S, and W; X ²⁰² is selected from C, D, F, G, L, M, Q, S, V, W, and ^Y ; is selected from A, C, D, E, L, M, N, S, W, and Y, X ²⁰⁴ is selected from A, D, I, M, V, and W, and X ²⁰⁵ is selected from D, E, G, and I, X ²⁰⁶ is selected from C, D, G, H, L, Q, S, and T, and X ²⁰⁷ is C, D, I, L, V, W, and Y, X ²⁰⁸ is selected from C, D, L, V, and W, and X ²⁰⁹ is selected from C, D, G, I, M, N, P, Q, and W , X ²¹⁰ is selected from C, F, L, M, P, S, T, and Y, X ²¹¹ is selected from C, F, L, V, and W, and X ²¹² is selected from L, IL-2Rβ ligand according to aspect 74A, selected from N, S, T, and V

Embodiment 77A. An IL-2Rβ ligand according to aspect 76A, wherein X ²⁰¹ is selected from C, F, L, S, and W.

Embodiment 78A. An IL-2Rβ ligand according to any one of aspects 76A-77A, wherein X ²⁰² is selected from C, D, F, G, L, M, Q, S, V, W, and Y.

Embodiment 79A. An IL-2Rβ ligand according to any one of aspects 76A-78A, wherein X ²⁰³ is selected from D and E.

Embodiment 80A. An IL-2Rβ ligand according to any one of aspects 76A-79A, wherein X ²⁰⁴ is V.

Embodiment 81A. An IL-2Rβ ligand according to any one of aspects 76A-80A, wherein X ²⁰⁵ is G.

Embodiment 82A. An IL-2Rβ ligand according to any one of aspects 76A-81A, wherein X ²⁰⁶ is D.

Embodiment 83A. IL-2Rβ ligand according to any one of aspects 76A-82A, wherein X ²⁰⁷ is selected from I, W, and Y

Embodiment 84A. An IL-2Rβ ligand according to any one of aspects 76A-83A, wherein X ²⁰⁸ is selected from C, D, L, V, and W.

Embodiment 85A. An IL-2Rβ ligand according to any one of aspects 76A-84A, wherein X ²⁰⁹ is D.

Embodiment 86A. An IL-2Rβ ligand according to any one of aspects 76A-85A, wherein X ²¹⁰ is selected from D, F, L, M, P, S, T, and Y.

Embodiment 87A. An IL-2Rβ ligand according to any one of aspects 76A-86A, wherein X ²¹¹ is selected from C, F, L, V, and W.

Embodiment 88A. IL-2Rβ ligand according to any one of aspects 76A-87A, wherein X ²¹² is selected from L and V

Embodiment 89A. X ²⁰¹ is selected from amino acids, X ²⁰² is selected from amino acids, X ²⁰³ is selected from D and E, X ²⁰⁴ is V, X ²⁰⁵ is G, X ²⁰⁶ is D and X ²⁰⁷ is selected from I, Y, and W, X ²⁰⁸ is selected from amino acids, X ²⁰⁹ is D, X ²¹⁰ is selected from amino acids, and X ²¹¹ is selected from amino acids 76A, wherein X ²¹² is selected from I, L, M, V, F, Y, and W.

Embodiment 90A. An IL-2Rβ ligand according to aspect 76A, wherein the IL-2Rβ ligand comprises an amino acid sequence selected from any one of SEQ ID NOS:1028-1042.

Embodiment 91A. the IL-2Rβ ligand comprises an amino acid sequence selected from any one of SEQ ID NOs: 1028-1042, wherein the amino acid sequence is on the N-terminus, the C-terminus, or both the N-terminus and the C-terminus 90A, wherein the IL-2Rβ ligand according to aspect 90A ends with amino acids -GG-.

Embodiment 92A. The IL-2Rβ ligand comprises an amino acid sequence selected from any one of SEQ ID NOS: 1028-1042, each amino acid independently having the following conservative substitutions: alanine (A), glycine ( G) amino acids with small hydrophobic side chains, including proline (P), serine (S), or threonine (T); hydroxyl-containing side chains, including serine (S), threonine (T), or tyrosine (Y) amino acids with acidic side chains, including aspartic acid (D), or glutamic acid (E); histidine (H), asparagine (N), glutamine (Q), serine (S), threonine (T), or amino acids with polar neutral side chains, including tyrosine (Y); amino acids with basic side chains, including arginine (R), lysine (K), or histidine (H); isoleucine (I), leucine (L), Amino acids with large hydrophobic side chains including methionine (M), valine (V), phenylalanine (F), tyrosine (Y), or tryptophan (W); as well as phenylalanine (F), histidine (H), tryptophan (W ), or amino acids with aromatic side chains including tyrosine (Y).

Embodiment 93A. An IL-2Rβ ligand according to any one of aspects 1A-9A, wherein the IL-2Rβ ligand comprises an amino acid sequence selected from any one of SEQ ID NOs:1044-1050.

Embodiment 94A. The IL-2Rβ ligand comprises an amino acid sequence selected from any one of SEQ ID NOs: 1044-1050, wherein the amino acid sequence is on the N-terminus, the C-terminus, or both the N-terminus and the C-terminus 93A, wherein the IL-2Rβ ligand according to aspect 93A ends with amino acids -GG-.

Embodiment 95A. The IL-2Rβ ligand comprises an amino acid sequence selected from any one of SEQ ID NOS: 1044-1050, each amino acid independently having the following conservative substitutions: alanine (A), glycine ( G) amino acids with small hydrophobic side chains, including proline (P), serine (S), or threonine (T); hydroxyl-containing side chains, including serine (S), threonine (T), or tyrosine (Y) amino acids with acidic side chains, including aspartic acid (D), or glutamic acid (E); histidine (H), asparagine (N), glutamine (Q), serine (S), threonine (T), or amino acids with polar neutral side chains, including tyrosine (Y); amino acids with basic side chains, including arginine (R), lysine (K), or histidine (H); isoleucine (I), leucine (L), Amino acids with large hydrophobic side chains including methionine (M), valine (V), phenylalanine (F), tyrosine (Y), or tryptophan (W); as well as phenylalanine (F), histidine (H), tryptophan (W ), or amino acids with aromatic side chains including tyrosine (Y).

Embodiment 96A. the IL-2Rβ ligand comprises the amino acid sequence of formula (10) (SEQ ID NO:578);
-X ¹⁹¹ -X ¹⁹² -X ¹⁹³ -X ¹⁹⁴ -X ¹⁹⁵ -CX ^{196 -X} 197 ^-X 198 -X ¹⁹⁹ -X ²⁰⁰ -X ²⁰¹ -X ²⁰² -X ²⁰³ -CX ²⁰⁴ -X ²⁰⁵ - X ²⁰⁶ -X ²⁰⁷ -X ²⁰⁸ - (10)
wherein X ¹⁹¹ is selected from amino acids containing large hydrophobic or aromatic side chains, X ¹⁹² is selected from amino acids, and X ¹⁹³ is amino acids containing large hydrophobic or aromatic side chains X ¹⁹⁴ is selected from amino acids containing large hydrophobic or basic side chains, X ¹⁹⁵ is selected from amino acids containing acidic or small hydrophobic side chains, X ¹⁹⁶ is selected from amino acids containing large is selected from hydrophobic or basic side chains, X ¹⁹⁷ is selected from amino acids containing large hydrophobic side chains, X ¹⁹⁸ is selected from amino acids containing small hydrophobic side chains, X ¹⁹⁹ is selected from polar / is selected from amino acids containing neutral or basic side chains, X ²⁰⁰ is selected from amino acids containing large hydrophobic side chains, X ²⁰¹ is selected from amino acids containing small hydrophobic side chains, X ²⁰² is selected from amino acids containing acidic or polar/neutral side chains, X ²⁰³ is selected from amino acids containing large hydrophobic side chains, X ²⁰⁴ is selected from amino acids containing acidic side chains, X ²⁰⁵ is selected from amino acids containing large hydrophobic side chains, X ²⁰⁶ is selected from amino acids containing acidic or aromatic side chains, X ²⁰⁷ is selected from amino acids containing amino acids, X ²⁰⁸ is , an amino acid comprising an acidic side chain.

Embodiment 97A. X ¹⁹¹ is selected from F, H, I, L, M, V, W, and Y; X ¹⁹² is selected from amino acids; X ¹⁹³ is F, H, I, L, M, V, W , and Y, X ¹⁹⁴ is selected from F, I, L, M, V, W, Y, H, K, and R, and X ¹⁹⁵ is selected from D, E, A, G, P, S , and T, X ¹⁹⁶ is selected from F, I, L, M, V, W, Y, H, K, and R, and X ¹⁹⁷ is selected from F, I, L, M, V, W , and Y, X ¹⁹⁸ is selected from A, G, P, S, and T, and X ¹⁹⁹ is selected from H, N, Q, S, T, Y, H, K, and R , X ²⁰⁰ is selected from F, I, L, M, V, W, and Y, X ²⁰¹ is selected from A, G, P, S, and T, X ²⁰² is selected from D, E, H , N, Q, S, T, and Y, X ²⁰³ is selected from F, I, L, M, V, W, and Y, X ²⁰⁴ is selected from D and E, and X ²⁰⁵ is selected from F, I, L, M, V, W, and Y; X ²⁰⁶ is selected from D, E, F, H, I, L, M, V ^, W, and Y; is selected from amino acids and X ²⁰⁸ is selected from D and E. IL-2Rβ ligand according to aspect 96A.

Embodiment 98A. An IL-2Rβ ligand according to aspect 97A, wherein X ¹⁹¹ is selected from F, H, W, and Y.

Embodiment 99A. An IL-2Rβ ligand according to any one of aspects 97A-98A, wherein X ¹⁹¹ is W.

Embodiment 100A. An IL-2Rβ ligand according to any one of aspects 97A-99A, wherein ^X192 is selected from amino acids.

Embodiment 101A. An IL-2Rβ ligand according to any one of aspects 97A-100A, wherein X ¹⁹³ is selected from F, H, W, and Y.

Embodiment 102A. An IL-2Rβ ligand according to any one of aspects 97A-100A, wherein X ¹⁹³ is selected from F, W, and Y.

Embodiment 103A. An IL-2Rβ ligand according to any one of aspects 97A-102A, wherein X ¹⁹⁴ is selected from H, L, and Y.

Embodiment 104A. An IL-2Rβ ligand according to any one of aspects 97A-102A, wherein X ¹⁹⁴ is L.

Embodiment 105A. An IL-2Rβ ligand according to any one of aspects 97A-102A, wherein X ¹⁹⁴ is Y.

Embodiment 106A. An IL-2Rβ ligand according to any one of aspects 97A-105A, wherein X ¹⁹⁵ is selected from D and P.

Embodiment 107A. An IL-2Rβ ligand according to any one of aspects 97A-105A, wherein ^X195 is D.

Embodiment 108A. An IL-2Rβ ligand according to any one of aspects 97A-105A, wherein X ¹⁹⁵ is P.

Embodiment 109A. An IL-2Rβ ligand according to any one of aspects 97A-108A, wherein X ¹⁹⁶ is selected from H and W.

Embodiment 110A. An IL-2Rβ ligand according to any one of aspects 97A-108A, wherein X ¹⁹⁶ is H.

Embodiment 111A. An IL-2Rβ ligand according to any one of aspects 97A-108A, wherein X ¹⁹⁶ is W.

Embodiment 112A. An IL-2Rβ ligand according to any one of aspects 97A-111A, wherein X ¹⁹⁷ is M.

Embodiment 113A. An IL-2Rβ ligand according to any one of aspects 97A-112A, wherein X ¹⁹⁸ is A.

Embodiment 114A. 113. An IL-2Rβ ligand according to any one of aspects 97AA-113, wherein X ¹⁹⁹ is selected from H, K, R, and Q.

Embodiment 115A. An IL-2Rβ ligand according to any one of aspects 97A-113A, wherein X ¹⁹⁹ is Q.

Embodiment 116A. An IL-2Rβ ligand according to any one of aspects 97A-113A, wherein X ¹⁹⁹ is selected from H, K and R.

Embodiment 117A. IL-2Rβ ligand according to any one of aspects 97A-116A, wherein X ²⁰⁰ is selected from L and V

Embodiment 118A. An IL-2Rβ ligand according to any one of aspects 97A-116A, wherein X ²⁰⁰ is L.

Embodiment 119A. An IL-2Rβ ligand according to any one of aspects 97A-118A, wherein X ²⁰¹ is G.

Embodiment 120A. An IL-2Rβ ligand according to any one of aspects 97A-119A, wherein X ²⁰² is selected from D, E, and Q.

Embodiment 121A. An IL-2Rβ ligand according to any one of aspects 97A-119A, wherein X ²⁰² is E.

Embodiment 122A. An IL-2Rβ ligand according to any one of aspects 97A-121A, wherein X ²⁰³ is L.

Embodiment 123A. An IL-2Rβ ligand according to any one of aspects 97A-122A, wherein X ²⁰⁴ is selected from D and E.

Embodiment 124A. An IL-2Rβ ligand according to any one of aspects 97A-122A, wherein X ²⁰⁴ is D.

Embodiment 125A. An IL-2Rβ ligand according to any one of aspects 97A-124A, wherein X ²⁰⁵ is L.

Embodiment 126A. An IL-2Rβ ligand according to any one of aspects 97A-125A, wherein X ²⁰⁶ is selected from D and E.

Embodiment 127A. An IL-2Rβ ligand according to any one of aspects 97A-126A, wherein X ²⁰⁷ is selected from amino acids.

Embodiment 128A. An IL-2Rβ ligand according to any one of aspects 97A-127A, wherein X ²⁰⁸ is selected from D and E.

Embodiment 129A. X ¹⁹¹ is selected from F, H, W, and Y; X ¹⁹² is selected from amino ^acids ; X ¹⁹³ is selected from F, H, W, and Y; is selected from Y, X ¹⁹⁵ is selected from D and P, X ¹⁹⁶ is selected from H, R, and W, X ¹⁹⁷ is M, X ¹⁹⁸ is A, X ¹⁹⁹ is H, K, R, and Q, X ²⁰⁰ is selected from L and V, X ²⁰¹ is G, X ²⁰² is selected from D, E, and Q, X ²⁰³ is L, and X ²⁰⁴ is selected from D and E, X ²⁰⁵ is L, X ²⁰⁶ is selected from D, E, H, F, W, and Y, X ²⁰⁷ is selected from amino acids, X ²⁰⁸ is An IL-2Rβ ligand according to aspect 97A, which is selected from D and E.

Embodiment 130A. X ¹⁹¹ is selected from F, H, W and Y, X ¹⁹² is selected from amino acids, X ¹⁹³ is Y, X ¹⁹⁴ is selected from H, L and Y, X ¹⁹⁵ is D X ¹⁹⁶ is W X ¹⁹⁷ is M X ¹⁹⁸ is A X ¹⁹⁹ is Q X ²⁰⁰ is selected from L and V X ²⁰¹ is G X ²⁰² is selected from D, E, and Q, X ²⁰³ is L, X ²⁰⁴ is selected from D and E, X ²⁰⁵ is L, X ²⁰⁶ is selected from D and E, X ^96A , wherein 207 is selected from amino acids and X ²⁰⁸ is selected from D and E. IL-2Rβ ligand according to aspect 96A.

Embodiment 131A. X ¹⁹¹ is selected from F, H, W and Y, X ¹⁹² is selected from amino acids, X ¹⁹³ is Y, X ¹⁹⁴ is selected from H, L and Y, X ¹⁹⁵ is D X ¹⁹⁶ is H, X ¹⁹⁷ is M, X ¹⁹⁸ is A, X ¹⁹⁹ is Q, X ²⁰⁰ is selected from L and V, X ²⁰¹ is G, X ²⁰² is selected from D, E, and Q, X ²⁰³ is L, X ²⁰⁴ is selected from D and E, X ²⁰⁵ is L, X ²⁰⁶ is selected from D and E, X ^96A , wherein 207 is selected from amino acids and X ²⁰⁸ is selected from D and E. IL-2Rβ ligand according to aspect 96A.

Embodiment 132A. X ¹⁹¹ is selected from F, H, W and Y, X ¹⁹² is selected from amino acids, X ¹⁹³ is Y, X ¹⁹⁴ is selected from H, L and Y, X ¹⁹⁵ is D X ¹⁹⁶ is R, X ¹⁹⁷ is M, X ¹⁹⁸ is A, X ¹⁹⁹ is Q, X ²⁰⁰ is selected from L and V, X ²⁰¹ is G, X ²⁰² is selected from D, E, and Q, X ²⁰³ is L, X ²⁰⁴ is selected from D and E, X ²⁰⁵ is L, X ²⁰⁶ is selected from D and E, X ^96A , wherein 207 is selected from amino acids and X ²⁰⁸ is selected from D and E. IL-2Rβ ligand according to aspect 96A.

Embodiment 133A. X ¹⁹¹ is selected from F, H, W and Y, X ¹⁹² is selected from amino acids, X ¹⁹³ is Y, X ¹⁹⁴ is selected from H, L and Y, X ¹⁹⁵ is P X ¹⁹⁶ is W X ¹⁹⁷ is M X ¹⁹⁸ is A X ¹⁹⁹ is Q X ²⁰⁰ is selected from L and V X ²⁰¹ is G X ²⁰² is selected from D, E, and Q, X ²⁰³ is L, X ²⁰⁴ is selected from D and E, X ²⁰⁵ is L, X ²⁰⁶ is selected from D and E, X ^96A , wherein 207 is selected from amino acids and X ²⁰⁸ is selected from D and E. IL-2Rβ ligand according to aspect 96A.

Embodiment 134A. X ¹⁹¹ is selected from F, H, W and Y, X ¹⁹² is selected from amino acids, X ¹⁹³ is Y, X ¹⁹⁴ is selected from H, L and Y, X ¹⁹⁵ is D and X ¹⁹⁶ is W, X ¹⁹⁷ is M, X ¹⁹⁸ is A, X ¹⁹⁹ is selected from H, K, and R, X ²⁰⁰ is selected from L and V, X ²⁰¹ is G, X ²⁰² is selected from D, E, and Q, X ²⁰³ is L, X ²⁰⁴ is selected from D and E, X ²⁰⁵ is L, X ²⁰⁶ is D and E, X ²⁰⁷ is selected from amino acids, and X ²⁰⁸ is selected from D and E. IL-2Rβ ligand according to aspect 96A.

Embodiment 135A. X ¹⁹¹ is selected from F, H, W and Y, X ¹⁹² is selected from amino acids, X ¹⁹³ is Y, X ¹⁹⁴ is selected from H, L and Y, X ¹⁹⁵ is D X ¹⁹⁶ is W X ¹⁹⁷ is M X ¹⁹⁸ is A X ¹⁹⁹ is Q X ²⁰⁰ is selected from L and V X ²⁰¹ is G X ²⁰² is selected from D, E, and Q, X ²⁰³ is L, X ²⁰⁴ is selected from D and E, X ²⁰⁵ is L, X ²⁰⁶ is F, H, W, and Y 96A, wherein X ²⁰⁷ is selected from amino acids and X ²⁰⁸ is selected from D and E.

Embodiment 136A. X ¹⁹¹ is selected from A, D, E, F, G, H, I, K, L, N, M, P, Q, R, S, T, V, W, and Y, and X ¹⁹² is is selected from A, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, and Y, and X ¹⁹³ is A, C, D, F, G, H, I, L, M, N, P, R, S, T, V, W, and Y, and X ¹⁹⁴ is F, H, I, K, L, N, P, Q, R, S, T, V, W, and Y, and X ¹⁹⁵ is selected from A, D, E, F, G, H, K, L, M, N, P, Q, S, W, and Y; X ¹⁹⁶ is selected from A, E, F, G, H, Q, R, S, W, and Y; X ¹⁹⁷ is selected from A, D, E, F, I, K, L, M, N, Q, R, S, T, V, W, and Y, wherein X ¹⁹⁸ is A and X ¹⁹⁹ is A, D, H, K, L, N, P , Q, R, S, and Y ^; X ²⁰⁰ is selected from I, L, M, P, and V; X ²⁰¹ is selected from G, H, and W; , E, and Q, X ²⁰³ is L, X ²⁰⁴ is selected from A, D, E, H, I, L, T, V, and Y, and X ²⁰⁵ is selected from F, I, L, M, V, W, and Y, and X ²⁰⁶ is A, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, and Y, and X ²⁰⁷ is A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, and Y, and X ²⁰⁸ is selected from A, D, E, F, G, H, K, L, M, N, P, Q, R, S, T, V, W, and Y A IL-2Rβ ligand according to aspect 96A.

Embodiment 137A. X ¹⁹¹ is selected from F, H, W, and Y, and X ¹⁹² is selected from A, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, is selected from T, V, W, and Y; X ¹⁹³ is selected from F, H, W, and Y; X ¹⁹⁴ is selected from F, H, I, L, V, W, and Y; X ¹⁹⁵ is selected from D, E, and P; X ¹⁹⁶ is selected from F, H, R, S, W, and Y; X ¹⁹⁷ is selected from F, I, L, M, and V X ¹⁹⁸ is selected from A; X ¹⁹⁹ is selected from H, K, N, Q, and R; X ²⁰⁰ is selected from I, L, and V; X ²⁰¹ is G; X ²⁰² is selected from D, E, and Q; ^{X 203} is selected from F, I, L, M, V, and Y; X ²⁰⁴ is selected from D and E; Yes, X ²⁰⁶ is selected from D, E, N, and Q; X ²⁰⁷ is selected from A, D, E, F, H, I, L, M, N, Q, T, and W; An IL-2Rβ ligand according to aspect 136A, wherein X ²⁰⁸ is selected from D and E.

Embodiment 138A. X ¹⁹¹ is W and X ¹⁹² is from A, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, and Y X ¹⁹³ is selected from F, H, W, ^and Y; X ¹⁹⁴ is Y; X ¹⁹⁵ is selected from D, E, and P; X ¹⁹⁷ is selected from I and M; X ¹⁹⁸ is A; X ¹⁹⁹ is selected from K, Q, and R; X ²⁰⁰ is selected from I, L, and V; X ²⁰¹ is G, X ²⁰² is E, X ²⁰³ is L, X ²⁰⁴ is D, X ²⁰⁵ is L, X ²⁰⁶ is selected from D and E, X ²⁰⁷ is A, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, and Y, and X ²⁰⁸ is selected from D and E An IL-2Rβ ligand according to aspect 136A, wherein

Embodiment 139A. X ¹⁹¹ is selected from A, D, E, F, G, H, I, K, L, N, M, P, Q, R, S, T, V, W, and Y, and X ¹⁹² is is selected from A, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, and Y, and X ¹⁹³ is F, H, W, and Y; X ¹⁹⁴ is selected from F, H, L, ^{W, and Y; X 195 is} selected from D, E, and P; is selected from S, W, and Y, X ¹⁹⁷ is selected from F, I, L, M, and V, X ¹⁹⁸ is A, and X ¹⁹⁹ is from H, K, Q, N, and R is selected, X ²⁰⁰ is selected from I, L, and V, X ²⁰¹ is G, X ²⁰² is selected from D, E, and Q, X ²⁰³ is L, and X ²⁰⁴ is D and E, X ²⁰⁵ is selected from F, I, L, M, V, and W, X ²⁰⁶ is selected from D, E, F, I, L, M, V, W, and Y is selected, X ²⁰⁷ is selected from A, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, and Y, and X ²⁰⁸ is , D and E. IL-2Rβ ligand according to aspect 136A.

Embodiment 140A. In aspect 139A, wherein X ¹⁹¹ is selected from A, D, E, F, G, H, I, K, L, N, M, P, Q, R, S, T, V, W, and Y. An IL-2Rβ ligand as described.

Embodiment 141A. An IL-2Rβ ligand according to any one of aspects 139A-140A, wherein X ¹⁹¹ is selected from A, G, P, S, and T.

Embodiment 142A. An IL-2Rβ ligand according to any one of aspects 139A-140A, wherein X ¹⁹¹ is selected from F, H, I, L, M, V, W, and Y.

Embodiment 143A. An IL-2Rβ ligand according to any one of aspects 139A-140A, wherein X ¹⁹¹ is selected from F, H, W, and Y.

Embodiment 144A. Embodiments 139A- wherein X ¹⁹² is selected from A, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, and Y 143A. IL-2Rβ ligand according to any one of 143A.

Embodiment 145A. An IL-2Rβ ligand according to any one of aspects 139A-143A, wherein X ¹⁹² is selected from A, G, P, S, and T.

Embodiment 146A. An IL-2Rβ ligand according to any one of aspects 139A-143A, wherein X ¹⁹² is selected from F, H, I, L, M, V, W, and Y.

Embodiment 147A. An IL-2Rβ ligand according to any one of aspects 139A-146A, wherein X ¹⁹³ is selected from F, H, W, and Y.

Embodiment 148A. An IL-2Rβ ligand according to any one of aspects 139A-146A, wherein X ¹⁹³ is W.

Embodiment 149A. An IL-2Rβ ligand according to any one of aspects 139A-148A, wherein X ¹⁹⁴ is selected from F, H, L, W, and Y.

Embodiment 150A. An IL-2Rβ ligand according to any one of aspects 139A-148A, wherein X ¹⁹⁴ is selected from H, L, and Y.

Embodiment 151A. An IL-2Rβ ligand according to any one of aspects 139A-148A, wherein X ¹⁹⁴ is Y.

Embodiment 152A. An IL-2Rβ ligand according to any one of aspects 139A-151A, wherein X ¹⁹⁵ is selected from D, E, and P.

Embodiment 153A. An IL-2Rβ ligand according to any one of aspects 139A-151A, wherein X ¹⁹⁵ is D.

Embodiment 154A. An IL-2Rβ ligand according to any one of aspects 139A-151A, wherein X ¹⁹⁵ is P.

Embodiment 155A. An IL-2Rβ ligand according to any one of aspects 139A-154A, wherein X ¹⁹⁶ is selected from F, H, R, S, W, and Y.

Embodiment 156A. An IL-2Rβ ligand according to any one of aspects 139A-154A, wherein X ¹⁹⁶ is selected from H, R, and W.

Embodiment 157A. An IL-2Rβ ligand according to any one of aspects 139A-154A, wherein X ¹⁹⁶ is W.

Embodiment 158A. An IL-2Rβ ligand according to any one of aspects 139A-157A, wherein X ¹⁹⁷ is selected from F, I, L, M, and V.

Embodiment 159A. An IL-2Rβ ligand according to any one of aspects 139A-157A, wherein X ¹⁹⁷ is selected from I and M.

Embodiment 160A. An IL-2Rβ ligand according to any one of aspects 139A-157A, wherein X ¹⁹⁷ is M.

Embodiment 161A. An IL-2Rβ ligand according to any one of aspects 139A-160A, wherein X ¹⁹⁸ is A.

Embodiment 162A. An IL-2Rβ ligand according to any one of aspects 139A-161A, wherein X ¹⁹⁹ is selected from H, K, Q, N, and R.

Embodiment 163A. An IL-2Rβ ligand according to any one of aspects 139A-161A, wherein X ¹⁹⁹ is selected from H, K, and R.

Embodiment 164A. An IL-2Rβ ligand according to any one of aspects 139A-161A, wherein X ¹⁹⁹ is Q.

Embodiment 165A. An IL-2Rβ ligand according to any one of aspects 139A-164A, wherein X ²⁰⁰ is selected from I, L, and V.

Embodiment 166A. IL-2Rβ ligand according to any one of aspects 139A-164A, wherein X ²⁰⁰ is selected from L and V

Embodiment 167A. An IL-2Rβ ligand according to any one of aspects 139A-166A, wherein X ²⁰¹ is G.

Embodiment 168A. An IL-2Rβ ligand according to any one of aspects 139A-167A, wherein X ²⁰² is selected from D, E, and Q.

Embodiment 169A. An IL-2Rβ ligand according to any one of aspects 139A-167A, wherein X ²⁰² is E.

Embodiment 170A. An IL-2Rβ ligand according to any one of aspects 139A-169A, wherein X ²⁰³ is L.

Embodiment 171A. An IL-2Rβ ligand according to any one of aspects 139A-170A, wherein X ²⁰⁴ is selected from D and E.

Embodiment 172A. An IL-2Rβ ligand according to any one of aspects 139A-170A, wherein X ²⁰⁴ is D.

Embodiment 173A. An IL-2Rβ ligand according to any one of aspects 139A-171A, wherein X ²⁰⁵ is selected from F, I, L, M, V, and W.

Embodiment 174A. An IL-2Rβ ligand according to any one of aspects 139A-171A, wherein X ²⁰⁵ is L.

Embodiment 175A. An IL-2Rβ ligand according to any one of aspects 139A-174A, wherein X ²⁰⁶ is selected from D and E.

Embodiment 176A. An IL-2Rβ ligand according to any one of aspects 139A-174A, wherein X ²⁰⁶ is D.

Embodiment 177A. An IL-2Rβ ligand according to any one of aspects 139A-174A, wherein X ²⁰⁶ is selected from F, I, L, M, V, W, and Y.

Embodiment 178A. of aspects 139A-177A, wherein X ²⁰⁷ is selected from A, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, and Y IL-2Rβ ligand according to any one.

Embodiment 179A. An IL-2Rβ ligand according to any one of aspects 139A-177A, wherein X ²⁰⁷ is selected from A, G, P, S, and T.

Embodiment 180A. An IL-2Rβ ligand according to any one of aspects 139A-177A, wherein X ²⁰⁷ is selected from F, I, L, M, V, W, and Y.

Embodiment 181A. An IL-2Rβ ligand according to any one of aspects 139A-180A, wherein X ²⁰⁸ is selected from D and E.

Embodiment 182A. X ¹⁹¹ is selected from F, I, L, M, V, W, and Y, and X ¹⁹² is selected from A, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, and Y; X ¹⁹³ is selected from F, H, W, and Y; X ¹⁹⁴ is selected from H ^, L, and Y; is selected from D and P, X ¹⁹⁶ is selected from H, R, and W, X ¹⁹⁷ is selected from I and M, X ¹⁹⁸ is A, X ^{199 is} H, K, Q , and R, X ²⁰⁰ is selected from L and V, X ²⁰¹ is G, X ²⁰² is selected from D, E, and Q, X ²⁰³ is L, and X ²⁰⁴ is D and E, X ²⁰⁵ is L, X ²⁰⁶ is selected from D, E, F, I, L, M, V, W, and Y, and X ²⁰⁷ is A, D, E, IL according to aspect 139A, wherein is selected from F, G, H, I, L, M, N, P, Q, R, S, T, W, and Y, and X ²⁰⁸ is selected from D and E -2Rβ ligand.

Embodiment 183A. X ¹⁹¹ is selected from F, I, L, M, V, W, and Y, and X ¹⁹² is selected from A, D, E, F, G, H, I, K, L, M, N, P, is selected from Q, R, S, T, V, W, and Y, X ¹⁹³ is W, X ¹⁹⁴ is Y, X ¹⁹⁵ is selected from D and P, X ¹⁹⁶ is W; X ¹⁹⁷ is M, X ¹⁹⁸ is A, X ¹⁹⁹ is Q, X ¹⁹⁹ is selected from H, K, and R, X ²⁰⁰ is selected from L and V, X ²⁰¹ is G , X ²⁰² is E, X ²⁰³ is L, X ²⁰⁴ is selected from D and E, X ²⁰⁵ is L, X ²⁰⁶ is D, X ²⁰⁷ is A, D, Embodiment 139A wherein X 208 is selected from E, F, G, H, I, L, M, N, Q, P, R, S, T, V, W, and Y, and X ²⁰⁸ is selected from D and E IL-2Rβ ligand as described in .

Embodiment 184A. X ¹⁹¹ is selected from F, I, L, M, V, W, and Y, X ¹⁹³ is W, X ¹⁹⁴ is Y, X ¹⁹⁵ is selected from D and P, X ¹⁹⁶ is selected from H, R ^, and W; X ¹⁹⁷ is M; X ¹⁹⁸ is A; X ¹⁹⁹ is selected from H, K, Q, and R; selected, X ²⁰¹ is G, X ²⁰² is E, X ²⁰³ is L, X ²⁰⁴ is D, X ²⁰⁵ is L, X ²⁰⁶ is D, X ²⁰⁷ is A , D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, and Y, and X ²⁰⁸ is selected from D and E , an IL-2Rβ ligand according to aspect 139A.

Embodiment 185A. An IL-2Rβ ligand according to aspect 97A, wherein the IL-2Rβ ligand comprises an amino acid sequence selected from any one of SEQ ID NO:579-SEQ ID NO:808.

Embodiment 186A. IL-2Rβ ligand comprises an amino acid sequence selected from any one of SEQ ID NOs:579-808, wherein the amino acid sequence is on the N-terminus, the C-terminus, or both the N-terminus and the C-terminus 185A, wherein the IL-2Rβ ligand according to aspect 185A ends with amino acids -GG-.

Embodiment 187A. The IL-2Rβ ligand comprises an amino acid sequence selected from any one of SEQ ID NO:579-SEQ ID NO:808, each amino acid independently having the following conservative substitutions: alanine (A), glycine ( G) amino acids with small hydrophobic side chains, including proline (P), serine (S), or threonine (T); hydroxyl-containing side chains, including serine (S), threonine (T), or tyrosine (Y) amino acids with acidic side chains, including aspartic acid (D), or glutamic acid (E); histidine (H), asparagine (N), glutamine (Q), serine (S), threonine (T), or amino acids with polar neutral side chains, including tyrosine (Y); amino acids with basic side chains, including arginine (R), lysine (K), or histidine (H); isoleucine (I), leucine (L), Amino acids with large hydrophobic side chains including methionine (M), valine (V), phenylalanine (F), tyrosine (Y), or tryptophan (W); as well as phenylalanine (F), histidine (H), tryptophan (W ), or amino acids with aromatic side chains including tyrosine (Y).

Embodiment 188A. the IL-2β ligand comprises the amino acid sequence of formula (11) (SEQ ID NO:809);
-X ²¹¹ -X ²¹² -X ²¹³ -X ²¹⁴ -CX ²¹⁵ -X ^{216 -X} 217 -X 218 ^{-X 219} -X ²²⁰ -X ²²¹ -X ²²² -CX ²²³ -X ²²⁴ -X ²²⁵ - (11)
wherein X ²¹¹ is selected from amino acids, X ²¹² is selected from amino acids containing aromatic side chains, X ²¹³ is selected from amino acids containing large hydrophobic or aromatic side chains, and X ²¹⁴ is P, X ²¹⁵ is selected from amino acids containing aromatic side chains, X ²¹⁶ is selected from amino acids containing large hydrophobic side chains, X ²¹⁷ is A, and X ²¹⁸ is selected from basic side chains. or an amino acid containing a polar/neutral side chain, X ²¹⁹ is selected from an amino acid containing a large hydrophobic side chain, X ²²⁰ is G, and X ²²¹ is an acidic side chain or a polar/neutral side chain. selected from amino acids containing side chains, X ²²² is L, X ²²³ is D, X ²²⁴ is selected from amino acids containing large hydrophobic side chains, X ²²⁵ is selected from amino acids containing acidic side chains The IL-2β ligand of any one of aspects 1A-9A that is selected.

Embodiment 189A. X ²¹¹ is selected from amino acids; X ²¹² is selected from F, H, W, and Y; X ^{213 is} selected from F, H, I, L, M, V, W, and Y; is P, X ²¹⁵ is selected from F, H, W, and Y, X ²¹⁶ is selected from F, I, L, M, V, W, and Y, X ²¹⁷ is A, X ²¹⁸ is selected from K, R, H, N, Q, S, T, and Y; X ²¹⁹ is selected from F, I, L, M, V ^, W, and Y; and X ²²¹ is selected from D, E, H, N, Q, S, T, and Y, X ²²² is L, X ²²³ is D, and X ²²⁴ is F, I, L , M, V, W, and Y, and X ²²⁵ is selected from D and E. IL-2Rβ ligand according to aspect 188A.

Embodiment 190A. An IL-2Rβ ligand according to aspect 189A, wherein X ²¹¹ is selected from amino acids.

Embodiment 191A. An IL-2Rβ ligand according to any one of aspects 189A-190A, wherein X ²¹¹ is selected from H, K, and R.

Embodiment 192A. An IL-2Rβ ligand according to any one of aspects 189A-190A, wherein X ²¹¹ is selected from H and R.

Embodiment 193A. An IL-2Rβ ligand according to any one of aspects 189A-192A, wherein X ²¹² is selected from F, H, W, and Y.

Embodiment 194A. An IL-2Rβ ligand according to any one of aspects 189A-192A, wherein X ²¹² is W.

Embodiment 195A. An IL-2Rβ ligand according to any one of aspects 189A-194A, wherein X ²¹³ is selected from F, H, I, L, M, V, W, and Y.

Embodiment 196A. An IL-2Rβ ligand according to any one of aspects 189A-194A, wherein X ²¹³ is L.

Embodiment 197A. An IL-2Rβ ligand according to any one of aspects 189A-194A, wherein X ²¹³ is Y.

Embodiment 198A. An IL-2Rβ ligand according to any one of aspects 189A-197A, wherein X ²¹⁴ is P.

Embodiment 199A. An IL-2Rβ ligand according to any one of aspects 189A-198A, wherein X ²¹⁵ is selected from F, H, W, and Y.

Embodiment 200A. An IL-2Rβ ligand according to any one of aspects 189A-198A, wherein X ²¹⁵ is W.

Embodiment 201A. An IL-2Rβ ligand according to any one of aspects 189A-200A, wherein X ²¹⁶ is selected from F, I, L, M, V, W, and Y.

Embodiment 202A. An IL-2Rβ ligand according to any one of aspects 189A-200A, wherein X ²¹⁶ is M.

Embodiment 203A. An IL-2Rβ ligand according to any one of aspects 189A-202A, wherein X ²¹⁷ is A.

Embodiment 204A. An IL-2Rβ ligand according to any one of aspects 189A-203A, wherein X ²¹⁸ is selected from K, R, H, N, Q, S, T, and Y.

Embodiment 205A. An IL-2Rβ ligand according to any one of aspects 189A-203A, wherein X ²¹⁸ is selected from K and R.

Embodiment 206A. An IL-2Rβ ligand according to any one of aspects 189A-203A, wherein X ²¹⁸ is Q.

Embodiment 207A. An IL-2Rβ ligand according to any one of aspects 189A-206A, wherein X ²¹⁹ is selected from F, I, L, M, V, W, and Y.

Embodiment 208A. An IL-2Rβ ligand according to any one of aspects 189A-206A, wherein X ²¹⁹ is L.

Embodiment 209A. An IL-2Rβ ligand according to any one of aspects 189A-208A, wherein X ²²⁰ is G.

Embodiment 210A. An IL-2Rβ ligand according to any one of aspects 189A-209A, wherein X ²²¹ is selected from D, E, H, N, Q, S, T, and Y.

Embodiment 211A. An IL-2Rβ ligand according to any one of aspects 189A-209A, wherein X ²²¹ is E.

Embodiment 212A. An IL-2Rβ ligand according to any one of aspects 189A-211A, wherein X ²²² is L.

Embodiment 213A. An IL-2Rβ ligand according to any one of aspects 189A-212A, wherein X ²²³ is D.

Embodiment 214A. An IL-2Rβ ligand according to any one of aspects 189A-213A, wherein X ²²⁴ is selected from F, I, L, M, V, W, and Y.

Embodiment 215A. An IL-2Rβ ligand according to any one of aspects 189A-213A, wherein X ²²⁴ is L.

Embodiment 216A. An IL-2Rβ ligand according to any one of aspects 189A-215A, wherein X ²²⁵ is selected from D and E.

Embodiment 217A. X ²¹¹ is selected from H, K, and R, X ²¹² is W, X ²¹³ is Y, X ²¹⁴ is P, X ²¹⁵ is W, X ²¹⁶ is M, X ²¹⁷ is A, X ²¹⁸ is selected from N and Q, X ²¹⁹ is selected from L and V, X ²²⁰ is G, X ²²¹ is selected from E, D, and Q, X An IL-2Rβ ligand according to aspect 188A, wherein ²²² is L, X ²²³ is D, X ²²⁴ is selected from L and M, and X ²²⁵ is selected from D and E.

Embodiment 218A. X ²¹¹ is selected from A, D, E, G, H, L, M, N, Q, R, S, T, and V; X ²¹² is selected from C, F, W, and Y; ^{X 213} is selected from F, H, K, L, N, Q, R, S, W, and Y; X ²¹⁴ is P; X ²¹⁵ is selected from W and Y; is selected from F, I, K, L, M, R, S, T, and V, X ²¹⁷ is A and X ²¹⁸ is D, E, G, H, K, L, N, Q, R , S, and Y, X ²¹⁹ is selected from L, P, and V, X ²²⁰ is selected from G, H, and W, and X ²²¹ is selected from D, E, and Q. , X ²²² is selected from L and M, X ²²³ is D, X ²²⁴ is selected from L, M, Q, and V, and X ²²⁵ is A, D, E, F, G, H , L, N, Q, T, and V. IL-2Rβ ligand according to aspect 188A.

Embodiment 219A. X ²¹¹ is selected from H and R, X ²¹² is selected from F and W, X ²¹³ is selected from F, L, W, and Y, X ²¹⁴ is P, X ²¹⁵ is is selected from W and Y, X ²¹⁶ is selected from F, I, L, M, and V, X ²¹⁷ is A, and X ²¹⁸ is D, E, H, K, N, Q, and R X ²¹⁹ is selected from L and V, X ²²⁰ is G, X ²²¹ is selected from D, E, and Q, X ²²² is selected from L and M, X ²²³ is selected from An IL-2Rβ ligand according to aspect 218A, wherein D, X ²²⁴ is selected from L, M, and V, and X ²²⁵ is selected from D and E.

Embodiment 220A. X ²¹¹ is selected from H and R, X ²¹² is W, X ²¹³ is Y, X ²¹⁴ is P, X ²¹⁵ is W, X ²¹⁶ is M, X ²¹⁷ is A , X ²¹⁸ is Q, X ²¹⁹ is L, X ²²⁰ is G, X ²²¹ is Q, X ²²² is L, X ²²³ is D, X ²²⁴ is L An IL-2Rβ ligand according to aspect 218A, wherein X ²²⁵ is selected from D and E.

Embodiment 221A. X ²¹¹ is selected from H and R, X ²¹² is W, X ²¹³ is L, X ²¹⁴ is P, X ²¹⁵ is W, X ²¹⁶ is M, X ²¹⁷ is A , X ²¹⁸ is Q, X ²¹⁹ is L, X ²²⁰ is G, X ²²¹ is Q, X ²²² is L, X ²²³ is D, X ²²⁴ is L An IL-2Rβ ligand according to aspect 218A, wherein X ²²⁵ is selected from D and E.

Embodiment 222A. X ²¹¹ is selected from H and R, X ²¹² is W, X ²¹³ is Y, X ²¹⁴ is P, X ²¹⁵ is W, X ²¹⁶ is M, X ²¹⁷ is A and X ²¹⁸ is selected from K and R, X ²¹⁹ is L, X ²²⁰ is G, X ²²¹ is Q, X ²²² is L, X ²²³ is D, X An IL-2Rβ ligand according to aspect 218A, wherein ²²⁴ is L and X ²²⁵ is selected from D and E.

Embodiment 223A. An IL-2Rβ ligand according to aspect 188A, wherein the IL-2Rβ ligand comprises an amino acid sequence selected from any one of SEQ ID NO:810-SEQ ID NO:903.

Embodiment 224A. IL-2Rβ ligand comprises an amino acid sequence selected from any one of SEQ ID NOs:810-903, wherein the amino acid sequence is on the N-terminus, the C-terminus, or both the N-terminus and the C-terminus An IL-2Rβ ligand according to aspect 223A, ending with amino acids -GG-.

Embodiment 225A. The IL-2Rβ ligand comprises an amino acid sequence selected from any one of SEQ ID NO:810-SEQ ID NO:903, each amino acid independently having the following conservative substitutions: alanine (A), glycine ( G) amino acids with small hydrophobic side chains, including proline (P), serine (S), or threonine (T); hydroxyl-containing side chains, including serine (S), threonine (T), or tyrosine (Y) amino acids with acidic side chains, including aspartic acid (D), or glutamic acid (E); histidine (H), asparagine (N), glutamine (Q), serine (S), threonine (T), or amino acids with polar neutral side chains, including tyrosine (Y); amino acids with basic side chains, including arginine (R), lysine (K), or histidine (H); isoleucine (I), leucine (L), Amino acids with large hydrophobic side chains including methionine (M), valine (V), phenylalanine (F), tyrosine (Y), or tryptophan (W); as well as phenylalanine (F), histidine (H), tryptophan (W ), or amino acids with aromatic side chains including tyrosine (Y).

Embodiment 226A. The IL of aspect 1A, wherein the IL-2Rβ ligand comprises an amino acid sequence selected from any one of SEQ ID NOs:1-193, 578-903, and 1028-1043. -2Rβ ligand.

Embodiment 227A. An IL-2Rβ ligand according to any one of aspects 1A-226A, wherein the IL-2Rβ ligand does not comprise the amino acid sequences of SEQ ID NO:268 and SEQ ID NO:374.

Embodiment 228A. A compound comprising at least one IL-2Rβ ligand of any one of aspects 1A-227A.

Embodiment 229A. The compound according to aspect 228A, wherein the compound further comprises at least one IL-2Rγc ligand of any one of SEQ ID NOs:194-267 and SEQ ID NOs:904-1027.

Embodiment 230A. The compound of any one of aspects 228A-229A, wherein the compound comprises at least one IL-2Rγc ligand.

Embodiment 231A. A compound according to any one of aspects 228A-230A, wherein the compound comprises an IL-2Rγc ligand, a linker, and an IL-2Rβ ligand.

Embodiment 232A. A compound according to aspect 231A, wherein the linker comprises a peptide linker.

Embodiment 233A. A compound according to aspect 231A, wherein the C-terminus of the IL-2Rγc ligand is covalently attached to the linker and the C-terminus of the IL-2Rβ ligand is attached to the linker.

Embodiment 234A. A compound according to aspect 231A, wherein the N-terminus of the IL-2Rγc ligand is covalently attached to the linker and the C-terminus of the IL-2Rβ ligand is attached to the linker.

Embodiment 235A. A compound according to aspect 231A, wherein the C-terminus of the IL-2Rγc ligand is covalently attached to the linker and the N-terminus of the IL-2Rβ ligand is attached to the linker.

Embodiment 236A. A compound according to aspect 231A, wherein the N-terminus of the IL-2Rγc ligand is covalently attached to the linker and the N-terminus of the IL-2Rβ ligand is attached to the linker.

Embodiment 237A. A compound according to any one of aspects 228A-236A, wherein the compound is selected from peptides, conjugates, fusion proteins, and single-chain peptides.

Embodiment 238A. A compound according to any one of aspects 228A-237A, wherein the compound comprises at least one moiety configured to modify properties of the conjugate.

Embodiment 239A. The properties are water solubility, polarity, lipophilicity, pharmacokinetic profile, targeting, bioavailability, pH dependent binding, bioactivity, pharmacodynamics, cellular activity, metabolism, efficacy, cage (irreversible), and the aforementioned A compound according to aspect 238A selected from any combination.

Embodiment 240A. A compound according to any one of aspects 238A-239A, wherein at least one portion comprises a small molecule, polymer, peptide, or antibody.

Embodiment 241A. A compound according to any one of aspects 228A-240A, which comprises a pharmacokinetic moiety.

Embodiment 242A. A compound according to aspect 241A, wherein the pharmacokinetic moiety comprises polyethylene glycol.

Embodiment 243A. A compound according to any one of aspects 228A-242A, which comprises a tumor-targeting moiety.

Embodiment 244A. A compound according to aspect 243A, wherein the tumor-targeting moiety comprises a tumor-specific antibody, tumor-specific antibody fragment, tumor-specific protein, or tumor-specific peptide.

Embodiment 245A. A compound according to any one of aspects 228A-244A, comprising an immune cell targeting moiety.

Embodiment 246A. A compound according to any one of aspects 228A-245A, wherein the compound is an IL-2R agonist.

Embodiment 247A. A compound according to any one of aspects 228A-246A, wherein the compound is an IL-2R antagonist.

Embodiment 248A. A compound according to any one of aspects 228A-247A, wherein the compound comprises a fusion protein and the IL-2Rβ ligand binds to the fusion partner.

Embodiment 249A. A compound according to aspect 248A, wherein the fusion protein partner comprises an IgG molecule, an IgG FAb fragment, or an Fc fragment.

Embodiment 250A. A compound according to aspect 248A, wherein the protein fusion partner comprises IL-2, a variant of IL-2, a mutant of IL-2, or an IL-2R agonist.

Embodiment 251A. A compound according to any one of aspects 228A-250A, wherein the compound comprises a label.

Embodiment 252A. A compound according to aspect 251A, wherein the label is selected from a radioisotope, a fluorophore, or a combination thereof.

Embodiment 253A. A compound according to any one of aspects 228A-252A, wherein the compound comprises a cage to protect peripheral tissue from the toxicity of IL-2R activation.

Embodiment 254A. A compound according to any one of aspects 228A-253A, wherein the compound comprises a moiety configured to target IL-2R-directed immune stimulation of effector immune cells in tumors.

Embodiment 255A. A compound according to any one of aspects 228A-254A, wherein the compound comprises a cleavable moiety.

Embodiment 256A. A compound according to any one of aspects 228A-240A, wherein the compound comprises a moiety that is toxic to cells expressing high levels of IL-2Rβ subunits.

Embodiment 257A. A compound according to aspect 256A, wherein the cells expressing high levels of IL-2Rβ subunits comprise cancer cells.

Embodiment 258A. A compound according to any one of aspects 228A-257A, wherein the compound comprises an imaging agent, a diagnostic agent, a targeting agent, a therapeutic agent, or a combination of any of the foregoing.

Embodiment 259A. A compound according to any one of aspects 228A-258A, wherein the compound comprises a moiety configured to target IL-2R-directed immune stimulation of effector immune cells in tumors.

Embodiment 260A. A pharmaceutical composition comprising an IL-2Rβ ligand according to any one of aspects 1A-227A, a compound according to any one of aspects 228A-259A, or a combination thereof.

Embodiment 261A. an IL-2Rγc ligand of any one of SEQ ID NOs: 194 to 267 and 904 to 1027, an IL-2Rγc ligand of any one of SEQ ID NOs: 194 to 267 and 904 to 1027 A pharmaceutical composition according to aspect 260A, further comprising a compound comprising, or a combination thereof.

Embodiment 262A. 260A. A method of treating cancer in a patient comprising administering a therapeutically effective amount of a pharmaceutical composition according to aspect 260A to a patient in need of such treatment.

Embodiment 263A. The method of aspect 262A, wherein the cancer comprises a solid tumor.

Embodiment 264A. A method of treating an autoimmune disease in a patient comprising administering to a patient in need of such treatment a therapeutically effective amount of a pharmaceutical composition according to any one of aspects 260A-261A. .

Embodiment 265A. A method of screening a compound for IL-2Rβ activity, wherein a cell is treated with an IL-2Rβ ligand according to any one of aspects 1A-227A, a compound according to any one of aspects 228A-259A, or the aforementioned wherein the cell expresses an IL-2Rβ subunit; contacting the cell with a test compound; determining the activity of the test compound; A method, including

Embodiment 266A. A method of activating the human IL-2 receptor, comprising activating a cell expressing the human IL-2 receptor in vivo with the IL-2Rβ ligand of any one of aspects 1A-227A and SEQ ID NO: 194 to number 267, and an IL-2Rγc ligand of any one of SEQ ID NOs:904-1027, or a compound of any one of aspects 228A-259A.

Embodiment 267A. A method of activating human IL-2 receptor in a patient, comprising treating a cell expressing human IL-2 receptor in vivo with the IL-2Rβ ligand of any one of aspects 1A-227A and SEQ ID NO: 194 SEQ ID NO:267, and any one of SEQ ID NOs:904-1027, or a compound of any one of aspects 228A-259A.

Embodiment 268A. A method of treating a disease in a patient, wherein the IL-2 receptor signaling pathway is implicated in disease etiology and the patient in need of such treatment is administered IL-2Rβ of any one of aspects 1A-227A. A method comprising administering a therapeutically effective amount of a compound comprising a ligand, or a compound of any one of aspects 228A-259A.

Embodiment 269A. A method of treating a disease in a patient, wherein activation of the IL-2 receptor is effective in treating the disease, and wherein the IL-2 receptor of any one of aspects 1A-227A is administered to the patient in need of such treatment. A method comprising administering a therapeutically effective amount of a compound comprising a 2Rβ ligand, or a compound of any one of aspects 228A-259A.

Embodiment 270A. A method of treating a disease in a patient, wherein inhibition of the IL-2 receptor is effective in treating the disease, wherein IL-2Rβ of any one of aspects 1A-227A is administered to the patient in need of such treatment. A method comprising administering a therapeutically effective amount of a compound comprising a ligand, or a compound of any one of aspects 228A-259A.

Aspect 271A A method of treating a disease in a patient, wherein cells expressing an IL-2Rβ subunit are implicated in the etiology of the disease, and wherein the patient in need of such treatment is treated with any one of aspects 1A-227A. A method comprising administering a therapeutically effective amount of a compound comprising an IL-2Rβ ligand, or a compound of any one of aspects 228A-259A.

Embodiment 272A. A method of treating a disease in a patient, wherein activation of IL-2R is effective in treating the disease, wherein the IL-2Rβ ligand of any one of aspects 1A-227A is administered to the patient in need of such treatment. or a compound of any one of aspects 228A-259A in a therapeutically effective amount.

Embodiment 273A. A method of treating a disease in a patient, wherein inhibition of IL-2R is effective in treating the disease, and the patient in need of such treatment is administered an IL-2Rβ ligand of any one of aspects 1A-227A. or a compound of any one of aspects 228A-259A in a therapeutically effective amount.

Embodiment 274A. A method of treating a disease in a patient, wherein reducing the sensitivity of Treg cells to IL-2 is effective in treating the disease, wherein the patient in need of such treatment is administered any one of aspects 1A-227A. A method comprising administering a therapeutically effective amount of a compound comprising an IL-2Rβ ligand, or a compound of any one of aspects 228A-259A.

Embodiment 275A. A method of imaging cells expressing IL-2Rβ subunits, wherein an effective amount of a compound comprising an IL-2Rβ ligand of any one of aspects 1A-227A or a compound of any one of aspects 228A-259A is administered to a patient. The above method, comprising administering to

Aspect 276A A method of diagnosing a disease in a patient, wherein the disease is associated with cells expressing IL-2Rβ subunits, and comprising an effective amount of an IL-2Rβ ligand according to any one of aspects 1A-227A. or administering to the patient a compound according to any one of aspects 228A-259A, and a compound comprising an IL-2Rβ ligand according to any one of aspects 1A-227A, or any of aspects 228A-259A determining the biodistribution of a compound according to one.

Embodiment 277A. A method of treating a disease in a patient, wherein a biological sample from the patient is treated with an IL-2Rβ ligand according to any one of aspects 1A-227A or an IL-2Rβ according to any one of aspects 1A-227A. contacting with a compound comprising a ligand; determining at least one property associated with binding of the IL-2Rβ ligand to cells of a biological sample; administering a compound comprising an IL-2Rβ ligand according to any one of aspects 1A-227A.

Embodiment 278A. The at least one property is the level of expression of the IL-2Rβ subunit in cells of the biological sample, a characterization of cells of the biological sample that express the IL-2Rβ subunit, and/or The method of aspect 277A, comprising cell binding affinity ( _IC50 ).

Embodiment 279A. A method of targeting a compound to cells expressing an IL-2Rβ subunit, comprising an effective amount of a compound comprising an IL-2Rβ ligand according to any one of aspects 1A-227A and a targeting moiety. A method comprising administering to a patient.

Embodiment 280A. A method of delivering a cytotoxic compound to a cell expressing an IL-2Rγc subunit, comprising an IL-2Rβ ligand according to any one of aspects 1A-227A, and a cytotoxic moiety, effective A method comprising administering an amount of a compound to a patient.

Embodiment 281A. The group of IL-2Rβ ligands having amino acid sequences of SEQ ID NOs: 1-163, 164-182, 578-808, and 1028-1043, the binding site of the IL-2Rβ subunit, are linked to other ILs within the group. - an IL-2Rβ ligand that competitively binds to the binding site with each of the 2Rβ ligands and has the amino acid sequence of SEQ ID NO: 1044 does not compete with the group of IL-2Rβ ligands for binding to the binding site, and IL-2 , binding sites for IL-2Rβ subunits that do not compete with the group of IL-2Rβ ligands for binding to the binding site.

Embodiment 282A. A binding site according to aspect 281A, wherein each IL-2Rβ ligand in the group of IL-2Rβ ligands has a binding affinity (IC ₅₀ ) for an IL-2Rβ subunit of less than 100 μM.

Embodiment 283A. A binding site according to any one of aspects 281A-282A, wherein each IL-2Rβ ligand of the group of IL-2Rβ ligands has a binding affinity (IC ₅₀ ) for the IL-2Rγc subunit of greater than 100 μM.

Embodiment 284A. A binding site according to any one of aspects 281A-283A, wherein the IL-2Rγc ligand having the amino acid sequence of SEQ ID NO:224 does not compete with the group of IL-2Rβ ligands for binding to the binding site.

Embodiment 285A. The binding site according to any one of aspects 281A-284A, wherein the group of IL-2Rβ ligands comprises IL-2Rβ ligands having the amino acid sequences of SEQ ID NOS:58, 83, 142, 169, 170 and 1042.

Aspect 1B. An IL-2Rγc ligand that exhibits a binding affinity (IC ₅₀ ) for the human IL-2Rγc subunit of less than 100 μM.

Aspect 2B. An IL-2Rγc ligand according to aspect 1B, wherein the IL-2Rγc ligand comprises 5-30 amino acids.

Aspect 3B. An IL-2Rγc ligand according to any one of aspects 1B-2B, wherein the IL-2Rγc ligand exhibits a binding affinity (IC ₅₀ ) for human IL-2Rγc subunits in the range of 1 pM to 100 μM.

Aspect 4B. An IL-2Rγc ligand according to any one of aspects 1B-2B, wherein the IL-2Rγc ligand exhibits a binding affinity (IC ₅₀ ) for human IL-2Rγc subunits in the range of 0.1 μM to 50 μM.

Aspect 5B. An IL-2Rγc ligand according to any one of aspects 1B-2B, wherein the IL-2Rγc ligand exhibits a binding affinity (IC ₅₀ ) for human IL-2Rγc subunits of less than 100 μM.

Aspect 6B. The IL of any one of aspects 1B-2B, wherein the IL-2Rγc ligand exhibits a binding affinity (IC ₅₀ ) of less than 100 μM for each of human IL-2Rβ subunit and human IL-2Rγc subunit. -2Rγc ligand.

Aspect 7B. An IL-2Rγc ligand according to any one of aspects 1B-2B, wherein the IL-2Rγc ligand exhibits a binding affinity (IC ₅₀ ) for the human IL-2Rα subunit of greater than 100 μM.

Aspect 8B. An embodiment wherein the IL-2Rγc ligand exhibits a binding affinity (IC ₅₀ ) for the human IL-2Rγc subunit that is at least 10-fold greater than the binding affinity (IC ₅₀ ) of the IL-2Rγc ligand for the human IL-2Rα subunit. IL-2Rγc ligand according to any one of 1B-2B.

Embodiment 9B. the IL-2Rγc ligand comprises the amino acid sequence of formula (4) (SEQ ID NO: 194) or the amino acid sequence of formula (4a) (SEQ ID NO: 195);
^{-X53 -X54 -X55 -X56 -X57 -X58 -X59 -X60} - (4)
^{-X51 -X52 -CX53 -X54 -X55 -X56 -X57 -X58 -X59 -X60 -CX61 -X62} - (4a)
wherein X ⁵¹ is selected from G, I, K, L, Q, R, T, Y, and V; X ⁵² is selected from A, D, E, H, I, L, M, R, S , T, V, and W, X ⁵³ is selected from D, E, F, N, Q, S, and T, and X ⁵⁴ is selected from A, D, E, G, I, M, N , Q, R, S, and T; X ⁵⁵ is selected from D, E, F, S, T, W, and Y; X ⁵⁶ is selected from D, E, F, G, L, M , N, Q, and Y; X ⁵⁷ is selected from E, G, N, S, and Q; X ⁵⁸ is selected from I, K, M, P, T, and V; ⁵⁹ is selected from I, L, M, S, T, and V ^; X ⁶⁰ is selected from F, I, and L; X ⁶¹ is selected from F, T, and W; , E, F, G, I, K, L, M, N, P, Q, S, V, W, and Y. .

Aspect 10B. An IL-2Rγc ligand according to aspect 9B, wherein ^X51 is selected from I, L, and V.

Aspect 11B. An IL-2Rγc ligand according to any one of aspects 9B-10B, wherein ^X52 is selected from S and T.

Aspect 12B. An IL-2Rγc ligand according to any one of aspects 9B-11B, wherein ^X53 is selected from D, E, N, and Q.

Aspect 13B. An IL-2Rγc ligand according to any one of aspects 9B-12B, wherein ^X54 is selected from D, E, N, and Q.

Aspect 14B. An IL-2Rγc ligand according to any one of aspects 9B-13B, wherein ^X55 is selected from F, W, and Y.

Aspect 15B. An IL-2Rγc ligand according to any one of aspects 9B-14B, wherein ^X56 is selected from D, E, N, and Q.

Aspect 16B. An IL-2Rγc ligand according to any one of aspects 9B-15B, wherein ^X57 is G.

Aspect 17B. An IL-2Rγc ligand according to any one of aspects 9B-16B, wherein ^X58 is selected from I and V.

Aspect 18B. An IL-2Rγc ligand according to any one of aspects 9B-17B, wherein ^X59 is selected from I, L, M, and V.

Aspect 19B. An IL-2Rγc ligand according to any one of aspects 9B-18B, wherein ^X60 is selected from F, I, and L.

Aspect 20B. An IL-2Rγc ligand according to any one of aspects 9B-19B, wherein ^X61 is W.

Aspect 21B. An IL-2Rγc ligand according to any one of aspects 9B-20B, wherein ^X62 is selected from N and Q.

Aspect 22B. X ⁵¹ is selected from I, L, and V; X ⁵² is selected from S ^and T; X ⁵³ is selected from D, E, N, and Q; , X ⁵⁵ is selected from F, W, and Y, X ⁵⁶ is selected from D, E, N, and Q, X ⁵⁷ is selected from G, and X ⁵⁸ is selected from I and V , X ⁵⁹ is selected from I, L, M, and V, X ⁶⁰ is selected from F, I, and L, X ⁶¹ is W, and X ⁶² is selected from N and Q The IL-2Rγc ligand according to aspect 9B, wherein

Aspect 23B. An IL-2Rγc ligand according to aspect 9B, wherein the IL-2Rγc ligand comprises an amino acid sequence selected from any one of SEQ ID NOs:196-210 and SEQ ID NOs:904-913.

Aspect 24B. The IL-2Rγc ligand comprises an amino acid sequence selected from any one of SEQ ID NOs:196-210 and SEQ ID NOs:904-913, wherein the amino acid sequence is on the N-terminus, on the C-terminus, or on the N-terminus The IL-2Rγc ligand according to aspect 23B, which terminates with amino acids -GG- on both the and C-terminus.

Aspect 25B. The IL-2Rγc ligand comprises an amino acid sequence selected from any one of SEQ ID NOs:196-210 and SEQ ID NOs:904-913, each amino acid independently having the following conservative substitution: alanine (A), amino acids with small hydrophobic side chains including glycine (G), proline (P), serine (S), or threonine (T); serine (S), threonine (T), or tyrosine (Y) amino acids with hydroxyl-containing side chains including; amino acids with acidic side chains including aspartic acid (D), or glutamic acid (E); histidine (H), asparagine (N), glutamine (Q), serine (S), Amino acids with polar neutral side chains, including threonine (T), or tyrosine (Y); amino acids with basic side chains, including arginine (R), lysine (K), or histidine (H); isoleucine (I) , leucine (L), methionine (M), valine (V), phenylalanine (F), tyrosine (Y), or tryptophan (W); and phenylalanine (F), histidine ( An IL-2Rγc ligand according to aspect 23B, comprising one or more of amino acids having aromatic side chains including H), tryptophan (W), or tyrosine (Y).

Aspect 26B. the IL-2Rγc ligand comprises the amino acid sequence of formula (4) (SEQ ID NO: 194) or the amino acid sequence of formula (4a) (SEQ ID NO: 195);
^{-X53 -X54 -X55 -X56 -X57 -X58 -X59 -X60} - (4)
^{-X51 -X52 -CX53 -X54 -X55 -X56 -X57 -X58 -X59 -X60 -CX61 -X62} - (4a)
wherein X ⁵¹ is selected from amino acids, X ⁵² is selected from amino acids, X ⁵³ is selected from amino acids containing polar neutral side chains or acidic side chains, X ⁵⁴ is selected from polar neutral side chains or selected from amino acids containing acidic side chains, X ⁵⁵ is selected from amino acids, X ⁵⁶ is selected from amino acids, X ⁵⁷ is selected from amino acids containing small hydrophobic side chains, X ⁵⁸ is selected from large is selected from amino acids containing a hydrophobic side chain, X ⁵⁹ is selected from amino acids containing a large hydrophobic side chain, X ⁶⁰ is selected from amino acids containing a large hydrophobic side chain, X ⁶¹ is selected from amino acids containing a large hydrophobic side chain An IL-2Rγc ligand according to any one of aspects 1B-8B, wherein ^X62 is selected from amino acids comprising side chains, and wherein X62 is selected from amino acids.

Aspect 27B. ^X51 is selected from amino acids containing large hydrophobic and basic side chains, ^X52 is selected from amino acids containing hydroxyl-containing and large hydrophobic side chains, and ^X53 is polar neutral. X ⁵⁴ is selected from amino acids containing a polar neutral side chain or an acidic side chain, X ⁵⁵ is selected from amino acids containing a large hydrophobic side chain, X ⁵⁶ is selected from amino acids containing polar neutral or acidic side chains, X ⁵⁷ is selected from amino acids containing small hydrophobic side chains, X ⁵⁸ is selected from amino acids containing large hydrophobic side chains, X ⁵⁹ is selected from amino acids with large hydrophobic side chains, X ⁶⁰ is selected from amino acids with large hydrophobic side chains, X ⁶¹ is selected from amino acids with large hydrophobic side chains, X ⁶² is selected from amino acids containing polar neutral side chains.

Aspect 28B. X ⁵¹ is selected from R, K, H, F, I, L, M, V, Y, and W, and X ⁵² is S, T, F, I, L, M, V, Y, and W with X ⁵³ selected from D, E, H, N, Q, S, T, and Y; X ⁵⁴ selected from D, E, H, N, Q, S, T, and Y , X ⁵⁵ is selected from F, I, L, M, V, Y, ^and W; X ⁵⁶ is selected from D, E, H, N, Q, S, T, and Y; is selected from A, G, P, S, and T; X ⁵⁸ is selected from F, I, L, M, V, Y, and W; X ⁵⁹ is selected from F, I, L, M, V, Y, and W; X ⁶⁰ is selected from F, I, L, M, V, Y, and W; X ⁶¹ is F, I, L, M, V, Y, and W and X ⁶² is selected from H, N, Q, S, and Y. An IL-2Rγc ligand according to any one of aspects 26B-27B.

Aspect 29B. An IL-2Rγc ligand according to aspect 28B, wherein ^X51 is selected from I, L, and V.

Aspect 30B. An IL-2Rγc ligand according to any one of aspects 28B-29B, wherein ^X52 is selected from S and T.

Embodiment 31B. An IL-2Rγc ligand according to any one of aspects 28B-30B, wherein ^X53 is selected from D, E, and Q.

Aspect 32B. An IL-2Rγc ligand according to any one of aspects 28B-31B, wherein ^X54 is selected from D, E, and N.

Aspect 33B. An IL-2Rγc ligand according to any one of aspects 28B-32B, wherein ^X55 is selected from F, Y, and W.

Aspect 34B. An IL-2Rγc ligand according to any one of aspects 28B-33B, wherein ^X56 is selected from D, E, N, and Q.

Aspect 35B. An IL-2Rγc ligand according to any one of aspects 28B-34B, wherein ^X57 is G.

Aspect 36B. An IL-2Rγc ligand according to any one of aspects 28B-35B, wherein ^X58 is selected from I and V.

Aspect 37B. An IL-2Rγc ligand according to any one of aspects 28B-36B, wherein ^X59 is selected from I, L, M, and V.

Aspect 38B. An IL-2Rγc ligand according to any one of aspects 28B-37B, wherein ^X60 is selected from F, I, and L.

Aspect 39B. An IL-2Rγc ligand according to any one of aspects 28B-38B, wherein ^X61 is W.

Aspect 40B. An IL-2Rγc ligand according to any one of aspects 28B-39B, wherein ^X62 is selected from N and Q.

Aspect 41B. X ⁵¹ is selected from I, L, and V; X ⁵² is selected from S ^and T; X ⁵³ is selected from D, E, and Q; X ⁵⁵ is selected from F, Y, and W; X ⁵⁶ is selected from D ^{, E, N, and Q; X 57 is} selected from G; X ⁵⁹ is selected from I, L, M, and V, X ⁶⁰ is selected from F, I, and L, X ⁶¹ is W, and X ⁶² is selected from N and Q IL-2Rγc ligand according to aspect 28B, wherein the IL-2Rγc ligand is

Aspect 42B. the IL-2Rγc ligand comprises the amino acid sequence of Formula (5) (SEQ ID NO:211) or Formula (5a) (SEQ ID NO:212);
^{-X73 -X74 -X75 -X76 -X77 -X78 -X79 -X80} - (5)
^{-X71 -X72 -CX73 -X74 -X75 -X76 -X77 -X78 -X79 -X80 -CX81 -X82} - (5a)
wherein X ⁷¹ is selected from F, G, I, L, P, Q, R, T, and V; and X ⁷² is selected from A, D, E, I, M, R, S, T, and V, X ⁷³ is selected from D, E, F, M, N, Q, ST, V, W, and Y, X ⁷⁴ is selected from D, E, F, G, I, L, M, P, R, S, T, and V; X ⁷⁵ is selected from F, H, L, W, and Y; X ⁷⁶ is D, E, H, L, N, Q, S, and T; ^{X 77} is selected from G, T, Q, and E; X ⁷⁸ is selected from I, L, M, Q, and V; N, Q, ^and R; X ⁸⁰ is selected from D, F, I, and L; X ⁸¹ is selected from F, I, L, R, T, W, and Y; is selected from A, F, G, H, I, L, N, P, Q, S, T, and W.

Aspect 43B. An IL-2Rγc ligand according to aspect 42B, wherein ^X71 is selected from I, L, and V.

Aspect 44B. An IL-2Rγc ligand according to any one of aspects 42B-43B, wherein ^X72 is selected from A, D, E, I, M, and V.

Aspect 45B. An IL-2Rγc ligand according to any one of aspects 42B-44B, wherein ^X73 is selected from E, Q, and N.

Aspect 46B. An IL-2Rγc ligand according to any one of aspects 42B-45B, wherein ^X74 is selected from D and E.

Aspect 47B. An IL-2Rγc ligand according to any one of aspects 42B-46B, wherein ^X75 is selected from F, W, and Y.

Aspect 48B. An IL-2Rγc ligand according to any one of aspects 42B-47B, wherein ^X76 is selected from D, E, L, N, and Q.

Aspect 49B. An IL-2Rγc ligand according to any one of aspects 42B-48B, wherein ^X77 is G.

Aspect 50B. An IL-2Rγc ligand according to any one of aspects 42B-49B, wherein ^X78 is selected from I, M, and V.

Embodiment 51B. An IL-2Rγc ligand according to any one of aspects 42B-50B, wherein ^X79 is selected from D, E, Q, and R.

Aspect 52B. An IL-2Rγc ligand according to any one of aspects 42B-51B, wherein ^X80 is selected from F, I, and L.

Aspect 53B. An IL-2Rγc ligand according to any one of aspects 42B-52B, wherein ^X81 is W.

Aspect 54B. An IL-2Rγc ligand according to any one of aspects 42B-53B, wherein ^X82 is selected from N and Q.

Aspect 55B. X ⁷¹ is selected from I, L, and V; X ⁷² is selected from A, D, E, I, M, and V; X ⁷³ is selected from E, Q, and ^N ; is selected from D and E, X ⁷⁵ is selected from F, W, and Y, X ⁷⁶ is selected from D, E, L, N, and Q, X ⁷⁷ is G, X ⁷⁸ is selected from I, M, and V; ^{X 79} is selected from D, E, Q, and R; X ⁸⁰ is selected from F, I, and L; , X ⁸² is selected from N and Q. The IL-2Rγc ligand according to aspect 42B.

Aspect 56B. the IL-2Rγc ligand comprises the amino acid sequence of Formula (5) (SEQ ID NO:211) or Formula (5a) (SEQ ID NO:212);
^{-X73 -X74 -X75 -X76 -X77 -X78 -X79 -X80} - (5)
^{-X71 -X72 -CX73 -X74 -X75 -X76 -X77 -X78 -X79 -X80 -CX81 -X82} - (5a)
wherein X ⁶¹ is selected from amino acids, X ⁶² is selected from amino acids, X ⁶³ is selected from amino acids, X ⁶⁴ is selected from amino acids, and X ⁶⁵ contains a large hydrophobic side chain is selected from amino acids, X ⁶⁶ is selected from amino acids, X ⁶⁷ is selected from small hydrophobic side chains, X ⁶⁸ is selected from amino acids containing large hydrophobic side chains, X ⁶⁹ is selected from basic side chains X ⁷⁰ is selected from amino acids containing large hydrophobic side chains, and X ⁷¹ is selected from amino acids containing large hydrophobic side chains. , ^X72 are selected from amino acids.

Aspect 57B. X ⁷¹ is selected from amino acids containing large hydrophobic side chains, X ⁷² is selected from amino acids containing acidic or large hydrophobic side chains, X ⁷³ is selected from acidic side chains, hydroxyl-containing side chains, or is selected from amino acids containing a polar neutral side chain, X ⁷⁴ is selected from amino acids containing an acidic side chain, a hydroxyl-containing side chain, or a large hydrophobic side chain, and X ⁷⁵ is an amino acid containing a large hydrophobic side chain wherein X ⁷⁶ is selected from an amino acid containing an acidic, hydroxyl-containing or polar neutral side chain, X ⁷⁷ is selected from a small hydrophobic side chain, X ⁷⁸ is a large hydrophobic is selected from amino acids containing side chains, X ⁷⁹ is selected from amino acids containing basic, acidic, or polar neutral side chains, and X ⁸⁰ is selected from amino acids containing large hydrophobic side chains; , X ⁸¹ is selected from amino acids containing large hydrophobic side chains, and X ⁸² is selected from amino acids containing polar neutral side chains.

Aspect 58B. X ⁷¹ is selected from F, I, L, M, V, Y, and W; X ⁷² is selected from D, E, F, I, L, M, V ^, Y, and W; is selected from D, E, S, T, H, N, Q, S, T, and Y, and X ⁷⁴ is selected from D, E, S, T, F, I, L, M, V, Y, and W, X ⁷⁵ is selected from F, I, L, M, V, Y, and W, X ⁷⁶ is selected from D, E, S, T, H, N, Q, S, T, and Y; X ⁷⁷ is selected from A, G, P, S, and T; X ⁷⁸ is selected from F, I, L, M, V, Y, and ^W ; R, K, H, D, E, H, N, Q, S, T, and Y; X ⁸⁰ is selected from F, I, L, M, V, Y, and W; X ⁸¹ is selected from F, I, L, M, V, Y, and W; and X ⁸² is selected from H, N, Q, S, T, and Y. An IL-2Rγc ligand as described in .

Aspect 59B. An IL-2Rγc ligand according to aspect 58B, wherein ^X71 is selected from I, L, and V.

Aspect 60B. An IL-2Rγc ligand according to any one of aspects 58B-59B, wherein ^X72 is selected from D, E, I, M, and V.

Embodiment 61B. An IL-2Rγc ligand according to any one of aspects 58B-60B, wherein ^X73 is selected from E, N, and Q.

Aspect 62B. An IL-2Rγc ligand according to any one of aspects 58B-61B, wherein ^X74 is selected from D and E.

Aspect 63B. An IL-2Rγc ligand according to any one of aspects 58B-62B, wherein ^X75 is selected from F, W, and Y.

Aspect 64B. An IL-2Rγc ligand according to any one of aspects 58B-63B, wherein ^X76 is selected from D, E, and N.

Aspect 65B. An IL-2Rγc ligand according to any one of aspects 58B-64B, wherein ^X77 is selected from G.

Aspect 66B. An IL-2Rγc ligand according to any one of aspects 58B-65B, wherein ^X78 is selected from I, M, and V.

Aspect 67B. An IL-2Rγc ligand according to any one of aspects 58B-66B, wherein ^X79 is selected from D, E, N, Q, and R.

Aspect 68B. An IL-2Rγc ligand according to any one of aspects 58B-67B, wherein ^X80 is selected from F, I, and L.

Aspect 69B. An IL-2Rγc ligand according to any one of aspects 58B-68B, wherein ^X81 is W.

Aspect 70B. An IL-2Rγc ligand according to any one of aspects 58B-69B, wherein ^X82 is selected from N and Q.

Embodiment 71B. X ⁷¹ is selected from I, L, and V; X ⁷² is selected from D, E, I, ^M , and V; X ⁷³ is selected from E, N, and Q; D and E, X ⁷⁵ is selected from F, W, and Y, X ⁷⁶ is selected from D, E, and N, X ⁷⁷ is selected from G, X ⁷⁸ is selected from I, M, and V; X ⁷⁹ is selected from D, E, N, Q, and R; X ⁸⁰ is selected from F, I, and L; X ⁸¹ is ^W ; is selected from N and Q. IL-2Rγc ligand according to aspect 58B.

Aspect 72B. An IL-2Rγc ligand according to aspect 56B, wherein the IL-2Rγc ligand comprises an amino acid sequence selected from any one of SEQ ID NOs:213-233 and SEQ ID NOs:914-920.

Aspect 73B. The IL-2Rγc ligand comprises an amino acid sequence selected from any one of SEQ ID NOs:213-233 and SEQ ID NOs:914-920, wherein the amino acid sequence is on the N-terminus, on the C-terminus, or on the N-terminus 72B, wherein the IL-2Rγc ligand according to aspect 72B ends with amino acids -GG- on both the and C-terminus.

Aspect 74B. The IL-2Rγc ligand comprises an amino acid sequence selected from any one of SEQ ID NOs:213-233 and SEQ ID NOs:914-920, each amino acid independently having the following conservative substitution: alanine (A), amino acids with small hydrophobic side chains including glycine (G), proline (P), serine (S), or threonine (T); serine (S), threonine (T), or tyrosine (Y) amino acids with hydroxyl-containing side chains including; amino acids with acidic side chains including aspartic acid (D), or glutamic acid (E); histidine (H), asparagine (N), glutamine (Q), serine (S), Amino acids with polar neutral side chains, including threonine (T), or tyrosine (Y); amino acids with basic side chains, including arginine (R), lysine (K), or histidine (H); isoleucine (I) , leucine (L), methionine (M), valine (V), phenylalanine (F), tyrosine (Y), or tryptophan (W); and phenylalanine (F), histidine ( An IL-2Rγc ligand according to aspect 72B, comprising one or more of amino acids having aromatic side chains including H), tryptophan (W), or tyrosine (Y).

Aspect 75B. the IL-2Rγc ligand comprises the amino acid sequence of Formula (6) (SEQ ID NO:234) or Formula (6a) (SEQ ID NO:235);
^{-X93 -X94 -X95 -X96 -X97 -X98 -X99 -X100 -X101} - (6)
^{-X91 -X92 -X93 -X94 -X95 -X96 -X97 -X98 -X99 -X100 -X101 -X102 -X103} - (6a)
wherein X ⁹¹ is selected from C, D, E, and L; X ⁹² is selected from C, L, M, R, S, V, and W; X ⁹³ is selected from C, D, F , P, and R, X ⁹⁴ is selected from A, D, L, Q, S, and W, X ⁹⁵ is selected from D, E, F, L, and V, and X ⁹⁶ is selected from , A, D, E, F, G, K, Q, and S, X ⁹⁷ is selected from E, L, M, and W, X ⁹⁸ is G, I, L, W, and Y, X ⁹⁹ is selected from E, I, R, T, and V, X ¹⁰⁰ is W, and X ¹⁰¹ is selected from C, A, I, L, P, and V , X ¹⁰² is selected from C, D, G, H, and X ¹⁰³ is selected from C, D, E, H, S, and T. IL-2Rγc ligand according to aspect 1B.

Aspect 76B. An IL-2Rγc ligand according to aspect 75B, wherein ^X91 is selected from D and E.

Aspect 77B. An IL-2Rγc ligand according to any one of aspects 75B-76B, wherein ^X92 is selected from L, M, R, S, V, and W.

Embodiment 78B. An IL-2Rγc ligand according to any one of aspects 75B-77B, wherein ^X93 is selected from D and F.

Aspect 79B. An IL-2Rγc ligand according to any one of aspects 75B-78B, wherein ^X94 is S.

Aspect 80B. An IL-2Rγc ligand according to any one of aspects 75B-79B, wherein ^X95 is selected from D and E.

Embodiment 81B. An IL-2Rγc ligand according to any one of aspects 75B-80B, wherein ^X96 is selected from D and E.

Embodiment 82B. An IL-2Rγc ligand according to any one of aspects 75B-81B, wherein ^X97 is selected from L, M, and W.

Aspect 83B. An IL-2Rγc ligand according to any one of aspects 75B-82B, wherein ^X98 is G.

Aspect 84B. An IL-2Rγc ligand according to any one of aspects 75B-83B, wherein ^X99 is E.

Embodiment 85B. An IL-2Rγc ligand according to any one of aspects 75B-84B, wherein X ¹⁰⁰ is W.

Embodiment 86B. An IL-2Rγc ligand according to any one of aspects 75B-85B, wherein X ¹⁰¹ is selected from I, L, and V.

Embodiment 87B. An IL-2Rγc ligand according to any one of aspects 75B-86B, wherein X ¹⁰² is selected from D and G.

Embodiment 88B. An IL-2Rγc ligand according to any one of aspects 75B-87B, wherein X ¹⁰³ is selected from S and T.

Embodiment 89B. X ⁹¹ is selected from D and E; X ⁹² is selected from L, M, R, S, V, and W; X ⁹³ is selected from D and F; X ⁹⁴ is S; X ⁹⁵ is selected from D and E, X ⁹⁶ is selected from D and E, X ⁹⁷ is selected from L, M and W, X ⁹⁸ is G and X ⁹⁹ is E and X ¹⁰⁰ is W, X ¹⁰¹ is selected from I, L, and V, X ¹⁰² is selected from D and G, and X ¹⁰³ is selected from S and T. IL-2Rγc ligands as described.

Aspect 90B. An IL-2Rγc ligand according to aspect 75B, wherein the IL-2Rγc ligand comprises an amino acid sequence selected from any one of SEQ ID NOs:236-245.

Embodiment 91B. the IL-2Rγc ligand comprises an amino acid sequence selected from any one of SEQ ID NOs:236-245, wherein the amino acid sequence is on the N-terminus, the C-terminus, or both the N-terminus and the C-terminus 90B, wherein the IL-2Rγc ligand according to aspect 90B ends with amino acids -GG-.

Embodiment 92B. The IL-2Rγc ligand comprises an amino acid sequence selected from any one of SEQ ID NO:236-SEQ ID NO:245, each amino acid independently having the following conservative substitutions: alanine (A), glycine ( G) amino acids with small hydrophobic side chains, including proline (P), serine (S), or threonine (T); hydroxyl-containing side chains, including serine (S), threonine (T), or tyrosine (Y) amino acids with acidic side chains, including aspartic acid (D), or glutamic acid (E); histidine (H), asparagine (N), glutamine (Q), serine (S), threonine (T), or amino acids with polar neutral side chains, including tyrosine (Y); amino acids with basic side chains, including arginine (R), lysine (K), or histidine (H); isoleucine (I), leucine (L), Amino acids with large hydrophobic side chains including methionine (M), valine (V), phenylalanine (F), tyrosine (Y), or tryptophan (W); as well as phenylalanine (F), histidine (H), tryptophan (W ), or amino acids with aromatic side chains including tyrosine (Y).

Aspect 93B. the IL-2Rγc ligand comprises the amino acid sequence of Formula (6) (SEQ ID NO:234) or Formula (6a) (SEQ ID NO:235);
^{-X93 -X94 -X95 -X96 -X97 -X98 -X99 -X100 -X101} - (6)
^{-X91 -X92 -X93 -X94 -X95 -X96 -X97 -X98 -X99 -X100 -X101 -X102 -X103} - (6a)
wherein X ⁹¹ is selected from amino acids containing acidic side chains or cysteine, X ⁹² is selected from amino acids, X ⁹³ is selected from amino acids containing acidic side chains or large hydrophobic side chains, and X ⁹⁴ is selected from amino acids, X ⁹⁵ is selected from amino acids, X ⁹⁶ is selected from amino acids, X ⁹⁷ is selected from amino acids containing large hydrophobic side chains, X ⁹⁸ is selected from small hydrophobic side chains or selected from amino acids with large hydrophobic side chains, X ⁹⁹ is selected from amino acids, X ¹⁰⁰ is selected from amino acids with large hydrophobic side chains, and X ¹⁰¹ is selected from amino acids with large hydrophobic side chains. wherein X ¹⁰² is selected from amino acids containing small hydrophobic or acidic side chains or cysteine and X ¹⁰³ is selected from amino acids containing acidic or hydroxyl-containing side chains or cysteine An IL-2Rγc ligand as described in .

Aspect 94B. X ⁹¹ is selected from amino acids containing acidic side chains, X ⁹² is selected from amino acids, X ⁹³ is selected from amino acids containing acidic side chains or large hydrophobic side chains, and X ⁹⁴ is selected from acidic side chains. or selected from amino acids containing hydroxyl-containing side chains, X ⁹⁵ is selected from amino acids containing acidic side chains, X ⁹⁶ is selected from amino acids, and X ⁹⁷ is selected from amino acids containing large hydrophobic side chains. , X ⁹⁸ is selected from amino acids containing small or large hydrophobic side chains, X ⁹⁹ is selected from amino acids containing acidic or large hydrophobic side chains, and X ¹⁰⁰ is selected from amino acids containing large hydrophobic side chains. is selected from amino acids containing side chains, X ¹⁰¹ is selected from amino acids containing large hydrophobic side chains, X ¹⁰² is selected from amino acids containing small hydrophobic or acidic side chains, and X ¹⁰³ is acidic An IL-2Rγc ligand according to aspect 93B, selected from amino acids containing side chains or hydroxyl-containing side chains.

Aspect 95B. X ⁹¹ is selected from D and E, X ⁹² is selected from amino acids, X ⁹³ is selected from D, E, F, I, L, M, V, Y, and W, X ⁹⁴ is D, E, S, and T; X ⁹⁵ is selected from D and E; X ⁹⁶ is selected from amino acids; X ⁹⁷ is F, I, L, M, V, Y, and W X ⁹⁸ is selected from A, G, P, S, T, F, I, L, M, V, Y, and W; X ⁹⁹ is selected from D, E, F, I, L, M, V, Y, and W; X ¹⁰⁰ is selected from F, I, L, M, V, Y, and W; X ¹⁰¹ is selected from F, I, L, M, V, Y, and W, X ¹⁰² is selected from D, E, A, G, P, S, and T, and X ¹⁰³ is selected from D, E, S, and T. An IL-2Rγc ligand according to any one.

Embodiment 96B. An IL-2Rγc ligand according to aspect 95B, wherein X ⁹¹ is selected from D and E.

Embodiment 97B. An IL-2Rγc ligand according to any one of aspects 95B-96B, wherein ^X92 is selected from amino acids.

Embodiment 98B. An IL-2Rγc ligand according to any one of aspects 95B-97B, wherein ^X93 is selected from D and F.

Aspect 99B. An IL-2Rγc ligand according to any one of aspects 95B-98B, wherein X ⁹⁴ is S.

Embodiment 100B. An IL-2Rγc ligand according to any one of aspects 95B-99B, wherein X ⁹⁵ is selected from D and E.

Aspect 101B. An IL-2Rγc ligand according to any one of aspects 95B-100B, wherein ^X96 is selected from amino acids.

Aspect 102B. An IL-2Rγc ligand according to any one of aspects 95B-101B, wherein X ⁹⁷ is selected from L, M, and W.

Aspect 103B. An IL-2Rγc ligand according to any one of aspects 95B-102B, wherein X ⁹⁸ is G.

Aspect 104B. An IL-2Rγc ligand according to any one of aspects 95B-103B, wherein X ⁹⁹ is E.

Aspect 105B. An IL-2Rγc ligand according to any one of aspects 95B-104B, wherein X ¹⁰⁰ is W.

Aspect 106B. An IL-2Rγc ligand according to any one of aspects 95B-105B, wherein X ¹⁰¹ is selected from I, L, and V.

Embodiment 107B. An IL-2Rγc ligand according to any one of aspects 95B-106B, wherein X ¹⁰² is selected from D and G.

Embodiment 108B. An IL-2Rγc ligand according to any one of aspects 95B-107B, wherein X ¹⁰³ is selected from S and T.

Aspect 109B. X ⁹¹ is selected from D and E, X ⁹² is selected from amino acids, X ⁹³ is selected from D and F, X ⁹⁴ is S, X ⁹⁵ is selected from D and E, X ⁹⁶ is selected from amino acids, X ⁹⁷ is selected from L, M and W, X ⁹⁸ is G, X ⁹⁹ is E, X ¹⁰⁰ is W, X ¹⁰¹ is , I, L, and V, X ¹⁰² is selected from D and G, and X ¹⁰³ is selected from S and T. IL-2Rγc ligand according to aspect 95B.

Embodiment 110B. the IL-2Rγc ligand comprises the amino acid sequence of Formula (7) (SEQ ID NO:246) or Formula (7a) (SEQ ID NO:247);
^{-X114 -X115 -CX116 -X117 -X118} - (7)
^{-X111 -X112 -X113 -X114 -X115 -CX116 -X117 -X118 -X119 -X120 -X121} - (7a)
wherein X ¹¹¹ is selected from D, G, I, and Q; X ¹¹² is selected from D, I, and L; X ¹¹³ is G, L, M, Q, R, S, and Y, X ¹¹⁴ is selected from D, E, G, L, S, T, and Y; X ¹¹⁵ is selected from E, L, P, and Q; X ¹¹⁶ is selected from D, E , K, L, S, and T, X ¹¹⁷ is selected from D, F, S, and W, X ¹¹⁸ is selected from F, N, W, and Y, and X ¹¹⁹ is selected from F , I, L, R, and W; X ¹²⁰ is selected from A, E, L, and S; and X ¹²¹ is selected from H, L, K, N, Q, and V; An IL-2Rγc ligand according to aspect 1B.

Embodiment 111B. An IL-2Rγc ligand according to aspect 110B, wherein X ¹¹¹ is selected from D and Q.

Embodiment 112B. An IL-2Rγc ligand according to any one of aspects 110B-111B, wherein X ¹¹² is selected from I and L.

Embodiment 113B. An IL-2Rγc ligand according to any one of aspects 110B-112B, wherein X ¹¹³ is selected from G, L, M, R, S, and Y.

Embodiment 114B. An IL-2Rγc ligand according to any one of aspects 110B-113B, wherein X ¹¹⁴ is L.

Embodiment 115B. An IL-2Rγc ligand according to any one of aspects 110B-114B, wherein X ¹⁵ is selected from E and Q.

Embodiment 116B. An IL-2Rγc ligand according to any one of aspects 110B-115B, wherein X ¹¹⁶ is selected from D and E.

Embodiment 117B. An IL-2Rγc ligand according to any one of aspects 110B-116B, wherein X ¹¹⁷ is selected from F and W.

Embodiment 118B. An IL-2Rγc ligand according to any one of aspects 110B-117B, wherein X ¹¹⁸ is selected from F, W, and Y.

Embodiment 119B. An IL-2Rγc ligand according to any one of aspects 110B-118B, wherein X ¹¹⁹ is selected from F, I, and L.

Aspect 120B. An IL-2Rγc ligand according to any one of aspects 110B-119B, wherein X ¹²⁰ is S.

Aspect 121B. An IL-2Rγc ligand according to any one of aspects 110B-120B, wherein X ¹²¹ is selected from N and Q.

Embodiment 122B. X ¹¹¹ is selected from D and Q, X ¹¹² is selected from I and L, X ¹¹³ is selected from G, L, M, R, S, and Y, X ¹¹⁴ is L, X ¹¹⁵ is selected from E and Q, X ¹¹⁶ is selected from D and E, X ¹¹⁷ is selected from F and W, X ¹¹⁸ is selected from F, W and Y, X ¹¹⁹ is , F, I, and L, X ¹²⁰ is S, and X ¹²¹ is selected from N and Q. IL-2Rγc ligand according to aspect 110B.

Aspect 123B. An IL-2Rγc ligand according to aspect 110B, wherein the IL-2Rγc ligand comprises an amino acid sequence selected from any one of SEQ ID NOs:248-254 and SEQ ID NOs:921-922.

Embodiment 124B. The IL-2Rγc ligand comprises an amino acid sequence selected from any one of SEQ ID NOs:248-254 and SEQ ID NOs:921-922, wherein the amino acid sequence is on the N-terminus, on the C-terminus, or on the N-terminus 123B, wherein the IL-2Rγc ligand according to aspect 123B ends with amino acids -GG- on both the and C-terminus.

Embodiment 125B. The IL-2Rγc ligand comprises an amino acid sequence selected from any one of SEQ ID NOs:248-254 and SEQ ID NOs:921-922, each amino acid independently having the following conservative substitution: alanine (A), amino acids with small hydrophobic side chains including glycine (G), proline (P), serine (S), or threonine (T); serine (S), threonine (T), or tyrosine (Y) amino acids with hydroxyl-containing side chains including; amino acids with acidic side chains including aspartic acid (D), or glutamic acid (E); histidine (H), asparagine (N), glutamine (Q), serine (S), Amino acids with polar neutral side chains, including threonine (T), or tyrosine (Y); amino acids with basic side chains, including arginine (R), lysine (K), or histidine (H); isoleucine (I) , leucine (L), methionine (M), valine (V), phenylalanine (F), tyrosine (Y), or tryptophan (W); and phenylalanine (F), histidine ( An IL-2Rγc ligand according to aspect 123B, comprising one or more of amino acids having aromatic side chains including H), tryptophan (W), or tyrosine (Y).

Embodiment 126B. the IL-2Rγc ligand comprises the amino acid sequence of Formula (7) (SEQ ID NO:246) or Formula (7a) (SEQ ID NO:247);
^{-X114 -X115 -CX116 -X117 -X118} - (7)
^{-X111 -X112 -X113 -X114 -X115 -CX116 -X117 -X118 -X119 -X120 -X121} - (7a)
wherein X ¹¹¹ is selected from amino acids, X ¹¹² is selected from amino acids containing large hydrophobic or acidic side chains, X ¹¹³ is selected from amino acids, and X ¹¹⁴ is acidic side chains or hydroxyl X ¹¹⁵ is selected from amino acids, X ¹¹⁶ is selected from amino acids, X ¹¹⁷ is selected from amino acids containing large hydrophobic side chains, and X ¹¹⁸ is selected from amino acids containing large hydrophobic side chains. X ¹¹⁹ is selected from amino acids containing a large hydrophobic side chain, X ¹²⁰ is selected from amino acids, and X ¹²¹ is selected from amino acids. IL-2Rγc ligand.

Embodiment 127B. X ¹¹¹ is selected from amino acids, X ¹¹² is selected from amino acids containing large hydrophobic or acidic side chains, X ¹¹³ is selected from amino acids, and X ¹¹⁴ is acidic or hydroxyl-containing side chains. X ¹¹⁵ is selected from amino acids containing a large hydrophobic side chain, X ¹¹⁶ is selected from amino acids containing an acidic side chain, and X ¹¹⁷ is selected from amino acids containing a large hydrophobic side chain X ¹¹⁸ is selected from amino acids containing large hydrophobic side chains, X ¹¹⁹ is selected from amino acids containing large hydrophobic side chains, X ¹²⁰ is selected from amino acids, X ¹²¹ is medium polar 125B, wherein the IL-2Rγc ligand according to aspect 125B is selected from amino acids comprising a positive side chain.

Embodiment 128B. X ¹¹¹ is selected from amino acids, X ¹¹² is selected from D, E, F, I, L, M, V, Y, and W, X ¹¹³ is selected from amino acids, X ¹¹⁴ is D, E, S, T, and Y; ^{X 115} is selected from F, I, L, M, V, Y, and W; X ¹¹⁶ is selected from D and E; , I, L, M, V, Y, and W, X ¹¹⁸ is selected from F, I, L, M, V, Y, and W, and X ¹¹⁹ is selected from F, I, L, M , V, Y, and W, X ¹²⁰ is selected from amino acids, and X ¹²¹ is selected from H, N, Q, S, T, and Y. An IL-2Rγc ligand as described in .

Embodiment 129B. An IL-2Rγc ligand according to aspect 128B, wherein X ¹¹¹ is selected from amino acids.

Aspect 130B. An IL-2Rγc ligand according to any one of aspects 128B-129B, wherein X ¹¹² is selected from I and L.

Aspect 131B. An IL-2Rγc ligand according to any one of aspects 128B-130B, wherein X ¹¹³ is selected from amino acids.

Aspect 132B. An IL-2Rγc ligand according to any one of aspects 128B-131B, wherein X ¹¹⁴ is selected from D, E, and S.

Aspect 133B. An IL-2Rγc ligand according to any one of aspects 128B-132B, wherein X ¹¹⁵ is L.

Embodiment 134B. An IL-2Rγc ligand according to any one of aspects 128B-133B, wherein X ¹¹⁶ is selected from D and E.

Embodiment 135B. An IL-2Rγc ligand according to any one of aspects 128B-134B, wherein X ¹¹⁷ is selected from F and W.

Embodiment 136B. An IL-2Rγc ligand according to any one of aspects 128B-135B, wherein X ¹¹⁸ is selected from F, W, and Y.

Embodiment 137B. An IL-2Rγc ligand according to any one of aspects 128B-136B, wherein X ¹¹⁹ is selected from F, I, and L.

Embodiment 138B. An IL-2Rγc ligand according to any one of aspects 128B-137B, wherein X ¹²⁰ is selected from amino acids.

Embodiment 139B. An IL-2Rγc ligand according to any one of aspects 128B-138B, wherein X ¹²¹ is selected from Q and N.

Aspect 140B. X ¹¹¹ is selected from amino acids, X ¹¹² is selected from I and L, X ¹¹³ is selected from amino acids, X ¹¹⁴ is selected from D, E and S, X ¹¹⁵ is selected from L , X ¹¹⁶ is selected from D and E, X ¹¹⁷ is selected from F and W, X ¹¹⁸ is selected from F, W, and Y, X ¹¹⁹ is selected from F, I, and L 128B, wherein X ¹²⁰ is selected from amino acids and X ¹²¹ is selected from Q and N.

Aspect 141B. An IL-2Rγc ligand according to aspect 126B, wherein the IL-2Rγc ligand comprises an amino acid sequence selected from any one of SEQ ID NOs:255-267 and SEQ ID NOs:923-930.

Embodiment 142B. The IL-2Rγc ligand comprises an amino acid sequence selected from any one of SEQ ID NOs:255-264 and SEQ ID NOs:923-930, wherein the amino acid sequence is on the N-terminus, on the C-terminus, or on the N-terminus An IL-2Rγc ligand according to aspect 141B, which terminates with amino acids -GG- on both the and C-terminus.

Aspect 143B. The IL-2Rγc ligand comprises an amino acid sequence selected from any one of SEQ ID NOs:255-264 and SEQ ID NOs:923-930, each amino acid independently having the following conservative substitution: alanine (A), amino acids with small hydrophobic side chains including glycine (G), proline (P), serine (S), or threonine (T); serine (S), threonine (T), or tyrosine (Y) amino acids with hydroxyl-containing side chains including; amino acids with acidic side chains including aspartic acid (D), or glutamic acid (E); histidine (H), asparagine (N), glutamine (Q), serine (S), Amino acids with polar neutral side chains, including threonine (T), or tyrosine (Y); amino acids with basic side chains, including arginine (R), lysine (K), or histidine (H); isoleucine (I) , leucine (L), methionine (M), valine (V), phenylalanine (F), tyrosine (Y), or tryptophan (W); and phenylalanine (F), histidine ( An IL-2Rγc ligand according to aspect 141B, comprising one or more of amino acids having aromatic side chains including H), tryptophan (W), or tyrosine (Y).

Embodiment 144B. the IL-2Rγc ligand comprises the amino acid sequence of formula (8) (SEQ ID NO:931);
^-CX131 -X132 ^{-X133 -X134 -X135 -X136 -X137 -X138 -X139 -X140 -X141 -X142 -C-} (8)
wherein X ¹³¹ is selected from amino acids with large hydrophobic side chains, X ¹³² is selected from amino acids with large hydrophobic side chains, and X ¹³³ is selected from amino acids with large hydrophobic side chains. , X ¹³⁴ is selected from amino acids containing large hydrophobic side chains, X ¹³⁵ is selected from amino acids containing basic side chains and acidic or polar neutral side chains, X ¹³⁶ is selected from amino acids, X ¹³⁷ is selected from amino acids containing small hydrophobic side chains, X ¹³⁸ is selected from amino acids containing acidic or polar neutral side chains, X ¹³⁹ is selected from amino acids containing large hydrophobic side chains, X ¹⁴⁰ is selected from amino acids containing small hydrophobic or hydroxyl-containing side chains, X ¹⁴¹ is selected from amino acids containing large hydrophobic side chains, and X ¹⁴² is selected from amino acids containing large hydrophobic side chains. The IL-2Rγc ligand according to aspect 1B, wherein:

Embodiment 145B. X ¹³¹ is selected from F, I, L, M, V, Y, and W; X ¹³² is selected from F, I, L, V, Y, and W; X ¹³³ is selected from F, I, L, M, V, Y, and W; X ¹³⁴ is selected from F, I, L, M, V, Y, and W; X ¹³⁵ is R, K, H, D, E, N, and Q, X ¹³⁶ is selected from amino acids, X ¹³⁷ is selected from A, G, P, S, and T, and X ¹³⁸ is selected from D, E, N, and Q , X ¹³⁹ is selected from F, I, L, M, V, Y, and W, X ¹⁴⁰ is selected from A, G, P, S, and T, and X ¹⁴¹ is selected from F, I, L , M, V, Y, and W, and X ¹⁴² is selected from F, I, L, M, V, Y, and W. IL-2Rγc ligand according to aspect 144B.

Embodiment 146B. X ¹³¹ is selected from G, G, A, E, F, G, L, and Y; X ¹³² is selected from I, S, N, L, I, and V; X ¹³³ is selected from A, M, L, Y, R, M, A, Y, and I; X ¹³⁴ is selected from Y, L, H, T, Y, F, K; X ¹³⁵ is R, Q, D , G, P, Y, L, I, K, and E, X ¹³⁶ is selected from S, G, T, I, F, Q, R, H, N, and L, and X ¹³⁷ is selected from , G, P, Q, T, D, G, N, and K, X ¹³⁸ is selected from E, D, K, F, T, X ¹³⁹ is selected from F, R, Y, V, L, and A; X ¹⁴⁰ is selected from T, W, N, E, D, S, T, and L; X ¹⁴¹ is selected from M, W, Y, F, A, and I and X ¹⁴² is selected from I, V, Y, L, Y, I, and E.

Embodiment 147B. An IL-2Rγc ligand according to aspect 146B, wherein X ¹³¹ is selected from F and Y.

Embodiment 148B. An IL-2Rγc ligand according to any one of aspects 146B-147B, wherein X ¹³² is selected from I, V, and L.

Embodiment 149B. An IL-2Rγc ligand according to any one of aspects 146B-147B, wherein X ¹³² is I.

Aspect 150B. An IL-2Rγc ligand according to any one of aspects 146B-149B, wherein X ¹³³ is selected from M, L, Y, and I.

Aspect 151B. An IL-2Rγc ligand according to any one of aspects 146B-150B, wherein X ¹³⁴ is selected from F, H, and Y.

Embodiment 152B. An IL-2Rγc ligand according to any one of aspects 146B-150B, wherein X ¹³⁴ is Y.

Aspect 153B. An IL-2Rγc ligand according to any one of aspects 146B-152B, wherein X ¹³⁵ is selected from R, K, D, and E.

Embodiment 154B. An IL-2Rγc ligand according to any one of aspects 146B-152B, wherein X ¹³⁵ is R.

Embodiment 155B. An IL-2Rγc ligand according to any one of aspects 146B-154B, wherein X ¹³⁶ is selected from amino acids.

Embodiment 156B. An IL-2Rγc ligand according to any one of aspects 146B-155B, wherein X ¹³⁷ is G.

Embodiment 157B. An IL-2Rγc ligand according to any one of aspects 146B-156B, wherein X ¹³⁸ is selected from D and E.

Embodiment 158B. An IL-2Rγc ligand according to any one of aspects 146B-156B, wherein X ¹³⁸ is E.

Embodiment 159B. An IL-2Rγc ligand according to any one of aspects 146B-158B, wherein X ¹³⁹ is selected from F, Y, and W.

Aspect 160B. An IL-2Rγc ligand according to any one of aspects 146B-158B, wherein X ¹³⁹ is F.

Aspect 161B. An IL-2Rγc ligand according to any one of aspects 146B-160B, wherein X ¹⁴⁰ is selected from S and T.

Embodiment 162B. An IL-2Rγc ligand according to any one of aspects 146B-161B, wherein X ¹⁴¹ is selected from F, I, L, M, V, Y, and W.

Aspect 163B. An IL-2Rγc ligand according to any one of aspects 146B-161B, wherein X ¹⁴¹ is Y.

Embodiment 164B. An IL-2Rγc ligand according to any one of aspects 146B-163B, wherein X ¹⁴² is selected from I, L, M, V, and Y.

Embodiment 165B. X ¹³¹ is selected from F and Y, X ¹³² is I, X ¹³³ is selected from M, L, Y, and I, X ¹³⁴ is Y, X ¹³⁵ is R, X ¹³⁶ is selected from amino acids, X ¹³⁷ is G, X ¹³⁸ is E, X ¹³⁹ is F, X ¹⁴⁰ is selected from S and T, X ¹⁴¹ is Y, X ¹⁴² is An IL-2Rγc ligand according to aspect 146B, which is selected from F, I, L, M, V, Y, and W.

Embodiment 166B. X ¹³¹ is F, X ¹³² is I, X ¹³⁴ is Y, X ¹²⁵ is R, X ¹³⁷ is G, X ¹³⁸ is E, X ¹³⁹ is F, X An IL-2Rγc ligand according to aspect 146B, wherein ¹⁴¹ is Y.

Embodiment 167B. An IL-2Rγc ligand according to aspect 144B, wherein the IL-2Rγc ligand comprises an amino acid sequence selected from any one of SEQ ID NOs:265-267 and SEQ ID NOs:932-940.

Embodiment 168B. The IL-2Rγc ligand comprises an amino acid sequence selected from any one of SEQ ID NOs:265-267 and SEQ ID NOs:932-940, wherein the amino acid sequence is on the N-terminus, on the C-terminus, or on the N-terminus 167B, wherein the IL-2Rγc ligand according to aspect 167B ends with amino acids -GG- on both the and C-terminus.

Embodiment 169B. The IL-2Rγc ligand comprises an amino acid sequence selected from any one of SEQ ID NOs:265-267 and SEQ ID NOs:932-940, each amino acid independently having the following conservative substitution: alanine (A), amino acids with small hydrophobic side chains including glycine (G), proline (P), serine (S), or threonine (T); serine (S), threonine (T), or tyrosine (Y) amino acids with hydroxyl-containing side chains including; amino acids with acidic side chains including aspartic acid (D), or glutamic acid (E); histidine (H), asparagine (N), glutamine (Q), serine (S), Amino acids with polar neutral side chains, including threonine (T), or tyrosine (Y); amino acids with basic side chains, including arginine (R), lysine (K), or histidine (H); isoleucine (I) , leucine (L), methionine (M), valine (V), phenylalanine (F), tyrosine (Y), or tryptophan (W); and phenylalanine (F), histidine ( An IL-2Rγc ligand according to aspect 167B, comprising one or more of amino acids having aromatic side chains including H), tryptophan (W), or tyrosine (Y).

Embodiment 170B. the IL-2Rγc ligand comprises the amino acid sequence of formula (9) (SEQ ID NO:941) or the amino acid sequence of formula (9a) (SEQ ID NO:942);
^{-X155 -X156 -X157 -X158 -X159} - (9)
^{-X151 -X152 -X153 -X154 -CX155 -X156 -X157 -X158 -X159 -CX160 -X161 -X162 -X163} - (9a)
wherein X ¹⁵¹ is selected from amino acids containing small hydrophobic side chains or hydroxyl-containing side chains, X ¹⁵² is selected from amino acids containing large hydrophobic side chains, and X ¹⁵³ is selected from acidic or polar neutral side chains. X ¹⁵⁴ is selected from amino acids containing a basic side chain, X ¹⁵⁵ is selected from amino acids containing a large hydrophobic side chain, X ¹⁵⁶ is selected from amino acids containing a small hydrophobic side chain or a hydroxyl containing side chains, X ¹⁵⁷ is selected from amino acids containing small hydrophobic side chains, X ¹⁵⁸ is selected from amino acids containing small hydrophobic or hydroxyl-containing side chains, and X ¹⁵⁹ is , amino acids containing small hydrophobic or hydroxyl-containing side chains, X ¹⁶⁰ is selected from amino acids containing small hydrophobic or hydroxyl-containing side chains, X ¹⁶¹ is selected from amino acids, X ¹⁶² is selected from amino acids containing large hydrophobic or basic side chains, and X ¹⁶³ is selected from amino acids containing large hydrophobic side chains.

Embodiment 171B. X ¹⁵¹ is selected from A, G, P, S, and T; X ¹⁵² is selected from F, I, L, M, V, Y, and W; X ¹⁵³ is selected from D, E, N, and Q, X ¹⁵⁴ is selected from H, K, and R, X ¹⁵⁵ is selected from F, I, L, M, V, Y, and W, and X ¹⁵⁶ is selected from A, G, is selected from P, S, T, and Y; X ¹⁵⁷ is selected from A, G, P, S, and T; X ¹⁵⁸ is selected from A, G, P, S, T, and Y; X ¹⁵⁹ is selected from A, G, P, S, T, and Y; X ¹⁶⁰ is selected from A, G, P, S, T, and Y; X ¹⁶¹ is selected from amino acids; Embodiment 170B, wherein ¹⁶² is selected from F, I, L, M, V, Y, W, R, K, and H, and X ¹⁶³ is selected from F, I, L, M, V, and W An IL-2Rγc ligand as described in .

Embodiment 172B. X ¹⁵¹ is selected from K, M, N, and K; X ¹⁵² is selected from M, L, ^and Y; X ¹⁵³ is selected from N, Y, and L; , X ¹⁵⁵ is selected from A, W, R, Y, and N, X ¹⁵⁶ is selected from T, N, and S, X ¹⁵⁷ is selected from P and A, X ¹⁵⁸ is selected from S, is selected from R, F, and L; X ¹⁵⁹ is selected from Q, S, E, ^{and T; X 160 is} selected from S, Q, and A; L, and N, X ¹⁶² is selected from I, K, R, and V, and X ¹⁶³ is selected from F and L- 2Rγc ligand.

Embodiment 173B. An IL-2Rγc ligand according to aspect 172B, wherein X ¹⁵¹ is selected from S and T.

Embodiment 174B. An IL-2Rγc ligand according to any one of aspects 172B-173B, wherein X ¹⁵² is selected from L and M.

Embodiment 175B. An IL-2Rγc ligand according to any one of aspects 172B-173B, wherein X ¹⁵² is L.

Embodiment 176B. An IL-2Rγc ligand according to any one of aspects 172B-175B, wherein X ¹⁵³ is N.

Embodiment 177B. An IL-2Rγc ligand according to any one of aspects 172B-176B, wherein X ¹⁵⁴ is K.

Embodiment 178B. An IL-2Rγc ligand according to any one of aspects 172B-177B, wherein X ¹⁵⁵ is selected from W and Y.

Embodiment 179B. An IL-2Rγc ligand according to any one of aspects 172B-178B, wherein X ¹⁵⁶ is selected from S and T.

Aspect 180B. An IL-2Rγc ligand according to any one of aspects 172B-178B, wherein X ¹⁵⁶ is S.

Embodiment 181B. An IL-2Rγc ligand according to any one of aspects 172B-180B, wherein X ¹⁵⁷ is P.

Embodiment 182B. An IL-2Rγc ligand according to any one of aspects 172B-181B, wherein X ¹⁵⁸ is S.

Embodiment 183B. An IL-2Rγc ligand according to any one of aspects 172B-182B, wherein X ¹⁵⁹ is selected from S and T.

Embodiment 184B. An IL-2Rγc ligand according to any one of aspects 172B-182B, wherein X ¹⁵⁹ is S.

Embodiment 185B. An IL-2Rγc ligand according to any one of aspects 172B-183B, wherein X ¹⁶⁰ is S.

Embodiment 186B. An IL-2Rγc ligand according to any one of aspects 172B-184B, wherein X ¹⁶¹ is selected from amino acids.

Embodiment 187B. An IL-2Rγc ligand according to any one of aspects 172B-185B, wherein X ¹⁶² is selected from I, V, R, and K.

Embodiment 188B. An IL-2Rγc ligand according to any one of aspects 172B-185B, wherein X ¹⁶² is selected from I and V.

Embodiment 189B. An IL-2Rγc ligand according to any one of aspects 172B-185B, wherein X ¹⁶² is selected from R and K.

Aspect 190B. An IL-2Rγc ligand according to any one of aspects 172B-186B, wherein X ¹⁶³ is selected from F and L.

Aspect 191B. An IL-2Rγc ligand according to any one of aspects 172B-186B, wherein X ¹⁶³ is L.

Aspect 192B. X ¹⁵¹ is selected from S and T, X ¹⁵² is L, X ¹⁵³ is N, X ¹⁵⁴ is K, X ¹⁵⁵ is selected from W and Y, X ¹⁵⁶ is S; X ¹⁵⁷ is P, X ¹⁵⁸ is S, X ¹⁵⁹ is S, X ¹⁶⁰ is S, T, X ¹⁶¹ is selected from amino acids, X ¹⁶² is I, X ¹⁶³ is An IL-2Rγc ligand according to aspect 172B, which is F.

Aspect 193B. X ¹⁵² is L, X ¹⁵³ is N, X ¹⁵⁴ is K, X ¹⁵⁶ is S, X ¹⁵⁷ is P, X ¹⁵⁸ is S, X ¹⁵⁹ is S, X An IL-2Rγc ligand according to aspect 172B, wherein ¹⁶⁰ is S, X ¹⁶² is I and X ¹⁶³ is F.

Embodiment 194B. An IL-2Rγc ligand according to aspect 170B, wherein the IL-2Rγc ligand comprises an amino acid sequence selected from any one of SEQ ID NOs:943-948.

Embodiment 195B. the IL-2Rγc ligand comprises an amino acid sequence selected from any one of SEQ ID NOs:943-948, wherein the amino acid sequence is on the N-terminus, the C-terminus, or both the N-terminus and the C-terminus 194B, wherein the IL-2Rγc ligand according to aspect 194B ends with amino acids -GG-.

Aspect 196B. The IL-2Rγc ligand comprises an amino acid sequence selected from any one of SEQ ID NOs:943-948, each amino acid independently having the following conservative substitutions: alanine (A), glycine ( G) amino acids with small hydrophobic side chains, including proline (P), serine (S), or threonine (T); hydroxyl-containing side chains, including serine (S), threonine (T), or tyrosine (Y) amino acids with acidic side chains, including aspartic acid (D), or glutamic acid (E); histidine (H), asparagine (N), glutamine (Q), serine (S), threonine (T), or amino acids with polar neutral side chains, including tyrosine (Y); amino acids with basic side chains, including arginine (R), lysine (K), or histidine (H); isoleucine (I), leucine (L), Amino acids with large hydrophobic side chains including methionine (M), valine (V), phenylalanine (F), tyrosine (Y), or tryptophan (W); as well as phenylalanine (F), histidine (H), tryptophan (W ), or amino acids with aromatic side chains including tyrosine (Y).

Embodiment 197B. the IL-2Rγc ligand comprises the amino acid sequence of formula (12) (SEQ ID NO:949);
-X ¹⁷¹ -X ¹⁷² -X ¹⁷³ -X ¹⁷⁴ -X ¹⁷⁵ -CX ^{176 -X} 177 ^-X 178 -X ¹⁷⁹ -X ¹⁸⁰ -X ¹⁸¹ -X ¹⁸² -X ¹⁸³ -CX ¹⁸⁴ -X ¹⁸⁵ - X ¹⁸⁶ -X ¹⁸⁷ -X ¹⁸⁸ -(12)
wherein X ¹⁷¹ is selected from amino acids containing a basic side chain, X ¹⁷² is selected from amino acids containing a hydroxyl-containing side chain, and X ¹⁷³ is selected from amino acids containing an acidic side chain or a large hydrophobic side chain. X ¹⁷⁴ is selected from amino acids containing a large hydrophobic side chain, X ¹⁷⁵ is selected from amino acids containing an acidic side chain or a large hydrophobic side chain, X ¹⁷⁶ is selected from an acidic side chain or a polar/medium X ¹⁷⁷ is selected from amino acids containing an acidic side chain, X ¹⁷⁸ is selected from amino acids containing a large hydrophobic or aromatic side chain, and X ¹⁷⁹ is selected from amino acids containing an acidic side chain. selected from amino acids containing side chains or polar/neutral side chains, X ¹⁸⁰ is G, X ¹⁸¹ is V, X ¹⁸² is E, X ¹⁸³ is L, X ¹⁸⁴ is W , X ¹⁸⁵ is selected from amino acids containing large hydrophobic side chains, X ¹⁸⁶ is E, X ¹⁸⁷ is selected from amino acids, and X ¹⁸⁸ is selected from amino acids containing acidic side chains. An IL-2Rγc ligand as described in .

Embodiment 198B. X ¹⁷¹ is selected from H, K, and R; X ¹⁷² is selected from S, T, and Y; X ¹⁷³ is selected from D, E, F, I, and ^V ; is selected from I and V, X ¹⁷⁵ is selected from E, I, L, M, and V, X ¹⁷⁶ is selected from D and E, X ¹⁷⁷ is selected from D and E, and X ¹⁷⁸ is , F and W, X ¹⁷⁹ is selected from D, E, N, and Q, X ¹⁸⁰ is G, X ¹⁸¹ is V, X ¹⁸² is selected from D and E, X ¹⁸³ is L, X ¹⁸⁴ is W, X ¹⁸⁵ is selected from I, L, Q, and V, X ¹⁸⁶ is selected from D and E, X ¹⁸⁷ is A, D, E, F, G, I, M, N, P, Q, R, S, T, V, W, and Y; and X ¹⁸⁸ is selected from D, E, N, and Q. IL-2Rγc ligands as described.

Embodiment 199B. An IL-2Rγc ligand according to aspect 198B, wherein X ¹⁷¹ is selected from H, K, and R.

Aspect 200B. An IL-2Rγc ligand according to any one of aspects 198B-199B, wherein X ¹⁷² is selected from S, T, and Y.

Aspect 201B. An IL-2Rγc ligand according to any one of aspects 198B-200B, wherein X ¹⁷³ is selected from D, E, F, I, L, M, V, W, and Y.

Aspect 202B. An IL-2Rγc ligand according to any one of aspects 198B-200B, wherein X ¹⁷³ is selected from D and E.

Aspect 203B. An IL-2Rγc ligand according to any one of aspects 198B-200B, wherein X ¹⁷³ is selected from F, I, L, M, V, W, and Y.

Aspect 204B. An IL-2Rγc ligand according to any one of aspects 198B-203B, wherein X ¹⁷⁴ is selected from F, I, L, M, V, W, and Y.

Aspect 205B. An IL-2Rγc ligand according to any one of aspects 198B-203B, wherein X ¹⁷⁴ is V.

Aspect 206B. An IL-2Rγc ligand according to any one of aspects 198B-205B, wherein X ¹⁷⁵ is selected from D, E, F, I, L, M, V, W, and Y.

Aspect 207B. An IL-2Rγc ligand according to any one of aspects 198B-205B, wherein X ¹⁷⁵ is selected from D and E.

Aspect 208B. An IL-2Rγc ligand according to any one of aspects 198B-205B, wherein X ¹⁷⁵ is selected from F, I, L, M, V, W, and Y.

Aspect 209B. An IL-2Rγc ligand according to any one of aspects 198B-208B, wherein X ¹⁷⁶ is selected from D, E, H, N, Q, S, T, and Y.

Aspect 210B. An IL-2Rγc ligand according to any one of aspects 198B-208B, wherein X ¹⁷⁶ is selected from E and Q.

Aspect 211B. An IL-2Rγc ligand according to any one of aspects 198B-210B, wherein X ¹⁷⁷ is selected from D and E.

Aspect 212B. An IL-2Rγc ligand according to any one of aspects 198B-211B, wherein X ¹⁷⁸ is selected from F, H, I, L, M, V, W, and Y.

Aspect 213B. An IL-2Rγc ligand according to any one of aspects 198B-211B, wherein X ¹⁷⁸ is selected from F, H, W, and Y.

Aspect 214B. An IL-2Rγc ligand according to any one of aspects 198B-211B, wherein X ¹⁷⁸ is W.

Aspect 215B. An IL-2Rγc ligand according to any one of aspects 198B-214B, wherein X ¹⁷⁹ is selected from D, E, H, N, Q, S, T, and Y.

Aspect 216B. An IL-2Rγc ligand according to any one of aspects 198B-214B, wherein X ¹⁷⁹ is selected from D, E, and Q.

Aspect 217B. An IL-2Rγc ligand according to any one of aspects 198B-216B, wherein X ¹⁸⁰ is G.

Aspect 218B. An IL-2Rγc ligand according to any one of aspects 198B-217B, wherein X ¹⁸¹ is V.

Aspect 219B. An IL-2Rγc ligand according to any one of aspects 198B-218B, wherein X ¹⁸² is E.

Aspect 220B. An IL-2Rγc ligand according to any one of aspects 198B-219B, wherein X ¹⁸³ is L.

Aspect 221B. An IL-2Rγc ligand according to any one of aspects 198B-220B, wherein X ¹⁸⁴ is W.

Aspect 222B. An IL-2Rγc ligand according to any one of aspects 198B-221B, wherein X ¹⁸⁵ is selected from F, I, L, M, V, W, and Y.

Aspect 223B. An IL-2Rγc ligand according to any one of aspects 198B-221B, wherein X ¹⁸⁵ is L.

Aspect 224B. An IL-2Rγc ligand according to any one of aspects 198B-223B, wherein X ¹⁸⁶ is E.

Aspect 225B. An IL-2Rγc ligand according to any one of aspects 198B-224B, wherein X ¹⁸⁷ is selected from amino acids.

Aspect 226B. An IL-2Rγc ligand according to any one of aspects 198B-225B, wherein X ¹⁸⁸ is selected from D and E.

Aspect 227B. X ¹⁷¹ is selected from H, K, and R, X ¹⁷² is selected from S, T, and Y, X ¹⁷³ is D, E, F, I, L, M, V, W, and Y with X ¹⁷⁴ selected from F, I, L, M, V, W, and Y; X ¹⁷⁵ selected from D, E, F, I, L, M, V, W, and Y , X ¹⁷⁶ is selected from D, E, H, N, Q, S, T, and Y, X ¹⁷⁷ is selected from D and E, and X ¹⁷⁸ is F, H, I, L, M , V, W, and Y, X ¹⁷⁹ is selected from D, E, H, N, Q, S, T, and Y, X ¹⁸⁰ is G, X ¹⁸¹ is V, and X ¹⁸² is E, X ¹⁸³ is L, X ¹⁸⁴ is selected from W, X ¹⁸⁵ is selected from F, L, M, V, W, and Y, X ¹⁸⁶ is E, X ¹⁸⁷ is selected from amino acids and X ¹⁸⁸ is selected from D and E. IL-2Rγc ligand according to aspect 198B.

Embodiment 228B. X ¹⁷¹ is selected from H, K, and R, X ¹⁷² is selected from S, T, and Y, X ¹⁷³ is D, E, F, I, L, M, V, W, and Y with X ¹⁷⁴ being V; X ¹⁷⁵ being selected from D, E, F, I, L, M, V, W, and Y; X ¹⁷⁶ being D, E, H, N, Q , S, T, and Y, X ¹⁷⁶ is selected from E and Q, X ¹⁷⁷ is selected from D and E, X ¹⁷⁸ is W, X ¹⁷⁹ is D, E, and Q wherein X ¹⁸⁰ is G, X ¹⁸¹ is V, X ¹⁸² is E, X ¹⁸³ is L, X ¹⁸⁴ is W, X ¹⁸⁵ is F, I, L, M 198B, wherein X ¹⁸⁶ is E, X ¹⁸⁷ is selected from amino acids, and X ¹⁸⁸ is selected from D and E.

Embodiment 229B. An IL-2Rγc ligand according to aspect 196B, wherein the IL-2Rγc ligand comprises the amino acid sequence of any one of SEQ ID NOs:950-1027.

Aspect 230B. the IL-2Rγc ligand comprises an amino acid sequence selected from any one of SEQ ID NOs:950 to 1027, wherein the amino acid sequence is on the N-terminus, the C-terminus, or both the N-terminus and the C-terminus 229B, wherein the IL-2Rγc ligand according to aspect 229B ends with amino acids -GG-.

Aspect 231B. The IL-2Rγc ligand comprises an amino acid sequence selected from any one of SEQ ID NOs:950-1027, each amino acid independently having the following conservative substitutions: alanine (A), glycine ( G) amino acids with small hydrophobic side chains, including proline (P), serine (S), or threonine (T); hydroxyl-containing side chains, including serine (S), threonine (T), or tyrosine (Y) amino acids with acidic side chains, including aspartic acid (D), or glutamic acid (E); histidine (H), asparagine (N), glutamine (Q), serine (S), threonine (T), or amino acids with polar neutral side chains, including tyrosine (Y); amino acids with basic side chains, including arginine (R), lysine (K), or histidine (H); isoleucine (I), leucine (L), Amino acids with large hydrophobic side chains including methionine (M), valine (V), phenylalanine (F), tyrosine (Y), or tryptophan (W); as well as phenylalanine (F), histidine (H), tryptophan (W ), or amino acids with aromatic side chains including tyrosine (Y).

Aspect 232B. An IL-2Rγc ligand according to aspect 1B, wherein the IL-2Rγc ligand comprises an amino acid sequence selected from any one of SEQ ID NOs:194-267 and SEQ ID NOs:904-1027.

Aspect 233B. An IL-2Rγc ligand according to any one of aspects 1B-232B, wherein the IL-2Rγc ligand does not comprise the amino acid sequence of SEQ ID NO:375 and SEQ ID NO:376.

Aspect 234B. A compound comprising at least one IL-2Rγc ligand of any one of embodiments 1B-233B.

Aspect 235B. The embodiment wherein the compound further comprises at least one IL-2Rβ ligand selected from any one of SEQ ID NOs:1-193, 578-903, and 1028-1043. 234B.

Aspect 236B. The compound of any one of aspects 234B-235B, wherein the compound comprises at least one IL-2Rγc ligand.

Aspect 237B. Embodiments wherein the compound comprises a linker, wherein the linker binds at least one IL-2Rγc ligand, at least one other IL-2Rγc ligand, at least one IL-2Rβ ligand, and/or at least one IL-2Rγc ligand A compound according to any one of 234B-235B.

Embodiment 238B. A compound according to any one of aspects 234B-235B, wherein the compound comprises an IL-2Rγc ligand, a linker, and an IL-2Rβ ligand.

Aspect 239B. A compound according to aspect 238B, wherein the linker comprises a peptide linker.

Aspect 240B. A compound according to any one of aspects 238B-239B, wherein the C-terminus of the IL-2Rγc ligand is covalently attached to the linker and the C-terminus of the IL-2Rβ ligand is attached to the linker.

Aspect 241B. A compound according to any one of aspects 238B-239B, wherein the N-terminus of the IL-2Rγc ligand is covalently attached to the linker and the C-terminus of the IL-2Rβ ligand is attached to the linker.

Aspect 242B. A compound according to any one of aspects 238B-239B, wherein the C-terminus of the IL-2Rγc ligand is covalently attached to the linker and the N-terminus of the IL-2Rβ ligand is attached to the linker.

Aspect 243B. A compound according to any one of aspects 238B-239B, wherein the N-terminus of the IL-2Rγc ligand is covalently attached to the linker and the N-terminus of the IL-2Rβ ligand is attached to the linker.

Aspect 244B. A compound according to any one of aspects 234B-243B, wherein the compound is selected from peptides, conjugates, fusion proteins, and single-chain peptides.

Aspect 245B. A compound according to any one of aspects 234B-244B, wherein the compound comprises at least one moiety configured to modify properties of the conjugate.

Aspect 246B. The properties are water solubility, polarity, lipophilicity, pharmacokinetic profile, targeting, bioavailability, pH dependent binding, bioactivity, pharmacodynamics, cellular activity, metabolism, efficacy, cage (irreversible), and the aforementioned A compound according to aspect 245B selected from any combination.

Aspect 247B. A compound according to any one of aspects 245B-246B, wherein at least one portion comprises a small molecule, polymer, peptide, or antibody.

Aspect 248B. A compound according to any one of aspects 234B-247B, which comprises a pharmacokinetic moiety.

Aspect 249B. A compound according to aspect 248B, wherein the pharmacokinetic moiety comprises polyethylene glycol.

Aspect 250B. A compound according to any one of aspects 234B-249B, comprising a tumor-targeting moiety.

Aspect 251B. A compound according to aspect 250B, wherein the tumor-targeting moiety comprises a tumor-specific antibody, tumor-specific antibody fragment, tumor-specific protein, or tumor-specific peptide.

Aspect 252B. A compound according to any one of aspects 234B-251B, comprising an immune cell targeting moiety.

Aspect 253B. A compound according to any one of aspects 234B-252B, wherein the compound is an IL-2R agonist.

Aspect 254B. A compound according to any one of aspects 234B-253B, wherein the compound is an IL-2R antagonist.

Aspect 255B. A compound according to any one of aspects 234B-254B, wherein the compound comprises a fusion protein and the IL-2Rα ligand binds to the fusion partner.

Aspect 256B. A compound according to aspect 255B, wherein the fusion protein partner comprises an IgG molecule, an IgG FAb fragment, or an Fc fragment.

Aspect 257B. A compound according to any one of aspects 255B-256B, wherein the protein fusion partner comprises IL-2, a variant of IL-2, a mutant of IL-2, or an IL-2R agonist.

Embodiment 258B. A compound according to any one of aspects 234B-257B, wherein the compound comprises a label.

Aspect 259B. A compound according to aspect 258B, wherein the label is selected from a radioisotope, a fluorophore, or a combination thereof.

Aspect 260B. A compound according to any one of aspects 234B-259B, wherein the compound comprises a cage to protect peripheral tissue from the toxicity of IL-2R activation.

Aspect 261B. A compound according to any one of aspects 234B-260B, wherein the compound comprises a moiety configured to target IL-2R-directed immune stimulation of effector immune cells in tumors.

Aspect 262B. A compound according to any one of aspects 234B-261B, wherein the compound comprises a cleavable moiety.

Aspect 263B. A compound according to any one of aspects 234B-262B, wherein the compound comprises a moiety that is toxic to cells expressing high levels of IL-2Rγc subunit.

Aspect 264B. A compound according to aspect 263B, wherein the cells expressing high levels of IL-2Rγc subunits comprise cancer cells.

Aspect 265B. A compound according to any one of aspects 234B-264B, wherein the compound comprises an imaging agent, a diagnostic agent, a targeting agent, a therapeutic agent, or a combination of any of the foregoing.

Aspect 266B. A compound according to any one of aspects 234B-265B, wherein the compound comprises a moiety configured to target IL-2R-directed immune stimulation of effector immune cells in tumors.

Aspect 267B. A pharmaceutical composition comprising an IL-2Rγc ligand according to any one of aspects 1B-233B, a compound according to any one of aspects 234B-266B, or a combination thereof.

Aspect 268B. an IL-2Rβ ligand selected from any one of SEQ ID NOs:1-193, SEQ ID NOs:578-903, and SEQ ID NOs:1028-1043; SEQ ID NOs:1-193, SEQ ID NOs: The pharmaceutical composition according to aspect 267B, further comprising a compound comprising an IL-2Rβ ligand selected from any one of SEQ ID NOs:578-903, and SEQ ID NOs:1028-1043; or combinations thereof.

Aspect 269B. A method of treating cancer in a patient comprising administering to a patient in need of such treatment a therapeutically effective amount of a pharmaceutical composition according to any one of aspects 267B-268B.

Aspect 270B. The method of aspect 269B, wherein the cancer comprises a solid tumor.

Aspect 271B. A method of treating an autoimmune disease in a patient comprising administering to the patient in need of such treatment a therapeutically effective amount of a pharmaceutical composition according to any one of aspects 267B-268B. .

Aspect 272B. A method of screening a compound for IL-2Rγc activity, wherein a cell is treated with an IL-2Rγc ligand according to any one of aspects 1B-233B, a compound according to any one of aspects 234B-266B, or the aforementioned wherein the cell expresses an IL-2Rγc subunit; contacting the cell with a test compound; determining the activity of the test compound; A method, including

Aspect 273B. A method of activating the human IL-2 receptor comprising treating a cell expressing the human IL-2 receptor in vivo with an IL-2Rγc ligand of any one of aspects 1B-233B and SEQ ID NOs: 1-193 , SEQ ID NOs:578-903, and SEQ ID NOs:1028-1043, or a compound of any one of aspects 234B-266B. including, method.

Aspect 274B. A method of activating human IL-2 receptor in a patient, comprising treating a cell expressing human IL-2 receptor in vivo with an IL-2Rγc ligand of any one of aspects 1B-233B and SEQ ID NO: 1 193, SEQ ID NOs:578-903, and SEQ ID NOs:1028-1043, or a compound of any one of aspects 234B-266B. method, including

Aspect 275B. A method of treating a disease in a patient, wherein the IL-2 receptor signaling pathway is implicated in the etiology of the disease and the patient in need of such treatment is administered IL-2Rγc of any one of aspects 1B-233B. A method comprising administering a therapeutically effective amount of a compound comprising a ligand, or a compound of any one of aspects 234B-266B.

Aspect 276B. A method of treating a disease in a patient, wherein activation of the IL-2 receptor is effective in treating the disease, and wherein the IL-2 receptor of any one of aspects 1B-233B is administered to the patient in need of such treatment. A method comprising administering a therapeutically effective amount of a compound comprising a 2Rγc ligand, or a compound of any one of aspects 234B-266B.

Embodiment 277B. A method of treating a disease in a patient, wherein inhibition of the IL-2 receptor is effective in treating the disease and administering to the patient in need of such treatment IL-2Rγc of any one of aspects 1B-233B. A method comprising administering a therapeutically effective amount of a compound comprising a ligand, or a compound of any one of aspects 234B-266B.

Embodiment 278B. A method of treating a disease in a patient, wherein cells expressing the IL-2Rγc subunit are implicated in the etiology of the disease, and wherein the patient in need of such treatment is administered the IL-2 of any one of aspects 1B-233B. A method comprising administering a therapeutically effective amount of a compound comprising a 2Rγc ligand, or a compound of any one of aspects 234B-266B.

Embodiment 279B. A method of treating a disease in a patient, wherein activation of IL-2R is effective in treating the disease, and wherein the IL-2Rγc ligand of any one of aspects 1B-233B is administered to the patient in need of such treatment. or a compound of any one of aspects 234B-266B in a therapeutically effective amount.

Aspect 280B. A method of treating a disease in a patient, wherein inhibition of IL-2R is effective in treating the disease, and wherein the patient in need of such treatment is administered an IL-2Rγc ligand of any one of aspects 1B-233B. or a compound of any one of aspects 234B-266B in a therapeutically effective amount.

Aspect 281B. A method of treating a disease in a patient, wherein reducing the sensitivity of Treg cells to IL-2 is effective in treating the disease, wherein the patient in need of such treatment is administered any one of aspects 1B-233B. A method comprising administering a therapeutically effective amount of a compound comprising an IL-2Rγc ligand, or a compound of any one of aspects 234B-266B.

Aspect 282B. A method of imaging cells expressing IL-2Rγc subunits, wherein an effective amount of a compound comprising an IL-2Rγc ligand of any one of aspects 1B-233B or a compound of any one of aspects 234B-266B is administered to a patient. The above method, comprising administering to

Aspect 283B. A method of diagnosing a disease in a patient, wherein the disease is associated with cells expressing an IL-2Rγc subunit and comprising an effective amount of an IL-2Rγc ligand according to any one of aspects 1B-233B, or aspect administering to the patient a compound according to any one of aspects 234B-266B and a compound comprising an IL-2Rγc ligand according to any one of aspects 1B-233B, or any one of aspects 234B-266B and determining the biodistribution of a compound according to A.

Embodiment 284B. A method of treating a disease in a patient, wherein a biological sample from the patient is treated with an IL-2Rγc ligand according to any one of aspects 1B-233B or an IL-2Rγc according to any one of aspects 1B-233B. contacting with a compound comprising a ligand; determining at least one property associated with binding of the IL-2Rγc ligand to cells of a biological sample; administering a compound comprising an IL-2Rγc ligand according to any one of aspects 1B-233B.

Embodiment 285B. The at least one characteristic is the level of expression of the IL-2Rγc subunit in cells of the biological sample, characterization of cells of the biological sample that express the IL-2Rγc subunit, and/or The method of aspect 284B, comprising cell binding affinity ( _IC50 ).

Aspect 286B. A method of targeting a compound to a cell expressing an IL-2Rγc subunit, comprising an effective amount of a compound comprising an IL-2Rγc ligand according to any one of aspects 1B-233B and a targeting moiety. A method comprising administering to a patient.

Embodiment 287B. A method of delivering a cytotoxic compound to a cell expressing an IL-2Rγc subunit, comprising an IL-2Rγc ligand according to any one of aspects 1B-233B, and a cytotoxic moiety, effective A method comprising administering an amount of a compound to a patient.

Embodiment 288B. IL-2Rγc subunit binding site, SEQ ID NOs: 194-210, 904-913, 211-233, 914-920, 234-245, 246-254, 921-922, 265-267, and 932-940 A group of IL-2Rγc ligands having the amino acid sequence of binds to the binding site competitively with each of the other IL-2Rγc ligands within the group, and an IL-2Rγc ligand having the amino acid sequence of SEQ ID NO:948 binds to IL - A binding site for an IL-2Rγc subunit that does not compete for binding to the binding site with the group of 2Rγc ligands and IL-2 does not compete with the group of IL-2Rγc ligands for binding to the binding site.

Embodiment 289B. A binding site according to aspect 288B, wherein each IL-2Rγc ligand of the IL-2Rγc ligand group has a binding affinity (IC ₅₀ ) for the IL-2Rγc subunit of less than 100 μM.

Aspect 290B. A binding site according to any one of aspects 288B-289B, wherein each IL-2Rγc ligand of the group of IL-2Rγc ligands has a binding affinity (IC ₅₀ ) for the IL-2Rβ subunit of greater than 100 μM.

Aspect 291B. A binding site according to any one of aspects 288B-290B, wherein the IL-2Rβ ligand having the amino acid sequence of SEQ ID NO:58 does not compete with the group of IL-2Rγc ligands for binding to the binding site.

Aspect 292B. The binding site according to any one of aspects 288B-291B, wherein the IL-2Rγc ligands consist of peptides having the amino acid sequences of SEQ ID NOs: 198, 202, 224, 236 and 248.

Finally, it should be noted that there are alternative ways of implementing the embodiments disclosed herein. Accordingly, the embodiments are illustrative and not limiting, and the claims are not to be limited to the details given herein, but are to be modified within their scope and equivalents. obtain.

Claims (13)

An IL-2Rγc ligand having the amino acid sequence of any one of SEQ ID NOS: 951-965, 967-969, 971-984, 986-1003, 1005-1008, and 1010-1027.

The IL-2Rγc ligand of claim 1 , wherein said IL-2Rγc ligand has the amino acid sequence of any one of SEQ ID NOs: 952-958, 961, 962, 965, 971, 972, 974, and 1012.

The IL-2Rγc ligand of claim 1 , wherein said IL-2Rγc ligand has the amino acid sequence of SEQ ID NO:965.

Each amino acid sequence independently has the following conservative substitutions:
amino acids with small hydrophobic side chains including alanine (A), glycine (G), proline (P), serine (S), or threonine (T);
amino acids with hydroxyl-containing side chains including serine (S), threonine (T), or tyrosine (Y);
amino acids with acidic side chains, including aspartic acid (D), or glutamic acid (E);
amino acids with polar neutral side chains including histidine (H), asparagine (N), glutamine (Q), serine (S), threonine (T), or tyrosine (Y);
amino acids with basic side chains including arginine (R), lysine (K), or histidine (H);
amino acids with large hydrophobic side chains including isoleucine (I), leucine (L), methionine (M), valine (V), phenylalanine (F), tyrosine (Y), or tryptophan (W); ) , histidine (H), tryptophan (W), or tyrosine ( Y ). ligand. The IL-2γc ligand of any one of claims 1-4 , wherein said IL-2Rγc ligand exhibits a binding affinity (IC ₅₀ ) for said human IL-2Rγc subunit of less than 100 μM. The IL-2Rγc ligand of any one of claims 1-5 , wherein said amino acid sequence ends with amino acids -GG- on the N-terminus, the C-terminus, or both the N-terminus and the C-terminus. . A compound comprising at least one IL-2Rγc ligand according to any one of claims 1-6 . 8. The compound of claim 7 , wherein said compound is selected from peptides, conjugates and fusion proteins. 9. The compound of claim 8 , wherein said compound comprises at least one IL-2Rβ ligand. 10. The compound of claim 9 , wherein said compound comprises said IL-2Rγc ligand, a linker and said IL-2Rβ ligand. 11. The compound of claim 10 , wherein said IL-2Rγc ligand binds to said IL-2Rβ ligand via said linker. A pharmaceutical composition comprising an IL-2Rγc ligand according to any one of claims 1-6 or a compound according to any one of claims 7-11 . 13. A pharmaceutical composition according to claim 12 for treating cancer, autoimmune diseases or inflammatory diseases.

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