Peptides with immunomodulatory properties

Abstract

Novel peptides with immunomodulatory activity in vitro and in vivo are provided. The peptides may contain specific striapathic regions of alternating hydrophilic and hydrophobic modules that can adopt an amphipathic conformation under physiological conditions. Peptides capable of specifically binding to key functional regions on one or more signaling proteins, specifically pro-inflammatory cytokines, macrophage inhibitory proteins, and histone regulatory proteins, are provided. These peptides are sufficiently stable in the circulation to allow intravenous administration. Pharmaceutical compositions containing the subject peptides are also provided. The subject peptides find use in methods for modulating macrophage activity. In some cases, the peptides are CD206-binding agents. Methods of treating subjects for conditions associated with chronic inflammation using the peptides and compositions of the present disclosure are also provided.

Description

[Background technology]

[0001] Acute inflammation is the initial tissue response to harmful stimuli. It involves a complex and highly regulated process that is initiated when cells present within injured tissue, including macrophages, dendritic cells, histiocytes, Kupffer cells, and mast cells, sense injury-associated molecules and become activated. Upon activation, these cells release inflammatory mediators, such as vasodilators, which induce increased blood flow and vascular permeability in the vicinity of the injury. This, in turn, leads to increased migration of plasma and leukocytes (including neutrophils and macrophages) from the blood to the injured tissue. Because inflammatory mediators are generally rapidly degraded, a constant stimulus is required for acute inflammation to persist. Consequently, acute inflammation ends when the harmful stimulus is removed.

[0002] Various agents, including but not limited to bacteria, viruses, physical injury, chemical injury, cancer, chemotherapy, and radiation therapy, can cause prolonged and excessive inflammation, depending on the specific agent and genetic makeup of the animal exposed. Such inflammation, known as chronic inflammation, is thought to be a contributing factor to many widespread and debilitating diseases, including heart disease, cancer, respiratory disease, stroke, neurological diseases such as Alzheimer's disease, diabetes, and kidney disease. The result of chronic inflammation is the destruction of normal tissue and its replacement with collagen-rich connective tissue. Collagen-rich connective tissue, also known as scar tissue, exhibits reduced tissue function compared to normal tissue. In turn, the persistent and long-term formation of scar tissue leads to fibrosis. Fibrosis is one of the common symptoms of diseases affecting the lungs, skin, liver, heart, and bone marrow, and is an important factor in diseases such as idiopathic pulmonary fibrosis, scleroderma, keloids, liver cirrhosis, myocardial fibrosis, diabetic kidney disease, myelodysplastic syndrome, and other disorders.

[0003] Studies of chronic inflammation and fibrosis have shown that, regardless of the activating agent and the tissue affected, a common network of signaling proteins tends to function together to establish a pro-inflammatory state. This network of signaling proteins includes several different cytokines, cytokine receptors, transcription factors, and microRNAs, including TGFβ, TGFβRII, and miRNA19b. Therefore, therapeutic agents that reduce inflammation without adverse side effects are of great interest. Summary of the Invention

[0004] Novel peptides are provided that have immunomodulatory activity in vitro and in vivo. The peptides may contain specific striapathic regions of alternating hydrophilic and hydrophobic modules that can adopt an amphipathic conformation under physiological conditions. The peptides can specifically bind to important functional regions on one or more signaling proteins, specifically pro-inflammatory cytokines, macrophage inhibitory proteins, and / or histone regulatory proteins. The present disclosure includes peptides that are sufficiently stable in the in vivo circulation after administration to a subject. Pharmaceutical compositions containing the subject peptides are also provided.

[0005] The subject peptides find use in methods for modulating macrophage activity. In some cases, the peptides are CD206 binding agents. Methods of treating subjects for conditions associated with chronic inflammation using the peptides and compositions of the present disclosure are also provided.

[0006] The features and advantages of the compositions and methods of the present invention are set forth or become more fully apparent in the following description and appended claims. For example, suitable immunomodulatory polypeptides can be identified by using the formulas and sequences described herein. Moreover, the features and advantages of the described compositions and methods can be learned by practice of the methods or will be obvious from the description. [Brief explanation of the drawings]

[0007] [Figure 1] 1 shows a graph of the results of reducing bleomycin-induced pulmonary fibrosis in a mouse model of pulmonary fibrosis. Fibrosis measurement is the Ashcroft score after trichrome staining. Collagen score is a quantitative measurement after hydroxyproline staining. Further details are provided in the Examples below. [Figure 2] In a mouse tumor inhibition model, exemplary peptides of interest are demonstrated to synergize with PD-1 checkpoint inhibitors to reduce tumor volume. Further details are provided in the Examples below. [Figure 3]

[0023] Figure 1 demonstrates that exemplary peptides RP832C and RP837 reduce macrophage viability in human scleroderma patient samples. Macrophage samples were evaluated after 96 hours of incubation with various concentrations of peptides. [Figure 4A] 1 shows the properties of peptides RP832C and RP837 in samples from healthy controls with low arginase:IFNg (interferon-gamma) ratios. [Figure 4B] 1 shows the characterization of peptides RP832C and RP837 in macrophage samples from scleroderma patients with high arginase:IFNg ratios. DETAILED DESCRIPTION OF THE INVENTION

[0008] The following description provides specific details to provide a thorough understanding of the present invention. However, well-known structures, materials, processes, techniques, and operations are not shown or described in detail to avoid obscuring aspects of the described immunomodulatory peptides and related methods of treating a subject. In addition, those skilled in the art will understand that the described immunomodulatory peptides and related methods of treating a subject can be implemented and used without these specific details. Indeed, the described immunomodulatory peptides and methods can be practiced by modifying the illustrated peptides, compositions, kits, and methods, and can be used in combination with other conventionally used methods, treatments, devices, and techniques.

[0009] Immunomodulatory Polypeptides As summarized above, the present disclosure provides immunomodulatory peptides, particularly peptides with immunosuppressive properties, and methods for administering such immunomodulatory peptides to subjects, particularly subjects suffering from or at risk of developing a medical condition associated with persistent or chronic inflammation. The terms "immunomodulatory" and "immunomodulation" are used interchangeably herein. In some cases, the immunomodulatory peptides described herein may be referred to as anti-inflammatory peptides, and vice versa. In certain cases, the immunomodulatory peptides (e.g., described herein) are anti-inflammatory peptides, e.g., the peptides have at least one anti-inflammatory property.

[0010] Particular embodiments of immunomodulatory polypeptides of interest that can be applied to or adapted for use with the peptides of the present disclosure are described by Jaynes et al. in WO2016 / 061133, the disclosure of which is incorporated herein by reference in its entirety.

[0011] The terms "peptide" and "polypeptide" are used interchangeably herein to refer to polymers constructed from amino acid residues. The term "amino acid residue," as used herein, refers to any naturally occurring amino acid, non-naturally occurring amino acid, or amino acid mimetic (such as a peptoid monomer). The amino acid residue may be in the L- or D-form.

[0012] The present disclosure includes immunomodulatory peptides having a striapathic region comprising at least 25% of the length of the polypeptide and at least one immunomodulatory property. The term "striapathic region" refers to a region or portion of a peptide sequence consisting of a sequence of alternating hydrophobic and hydrophilic modules. A "hydrophobic module" is a peptide sequence consisting of 1 to 5 (e.g., 1 to 3 or 1 to 2) hydrophobic amino acid residues, e.g., 1, 2, 3, 4, or 5 hydrophobic amino acid residues. A "hydrophilic module" is a peptide sequence consisting of 1 to 5 (e.g., 1 to 3 or 1 to 2) hydrophilic amino acid residues, e.g., 1, 2, 3, 4, or 5 hydrophilic amino acid residues.

[0013] Therefore, the striated region is defined by the formula (X 1-5 J 1-5 ) n or (J 1-5 X 1-5 ) nwhere each X represents a hydrophilic amino acid residue, each J represents a hydrophobic amino acid residue, and each n is an integer from 1 to 10, such as 2 to 10, 2 to 8, 3 to 8, 4 to 8, or 5 to 10. As described in further detail below, aspects of the present disclosure include immunomodulatory peptides having a stripathic region with a particular degree of cationic charge. The immunomodulatory peptides of the present disclosure may include a stripathic region with a cationic surface. In certain embodiments, the stripathic region has a cationic charge (i.e., a charge greater than 0, e.g., +1, +2, +3, +4, +5, +6, or more). In certain embodiments, the immunomodulatory peptides include a tail region (e.g., a hydrophobic tail sequence). In certain embodiments, the immunomodulatory peptides include two or more stripathic regions. In such embodiments, the two amphipathic regions of the peptide are in the form of a dimer, and the two amphipathic regions may have the same or different amino acid sequences (i.e., a homodimer or a heterodimer). In certain embodiments, the two (or more) stripathic regions are connected via a linker or linking region. The linker may be a contiguous (or in-line) amino acid sequence or a non-amino acid moiety, as desired.

[0014] Hydrophobic amino acid residues are characterized by side chain groups with predominantly non-polar chemical or physical properties in, for example, physiological conditions, where the peptide finds use. Such hydrophobic amino acid residues may be naturally occurring or non-naturally occurring. Hydrophobic amino acid residues may be mimetics of naturally occurring amino acids with side chain groups that have predominantly non-polar chemical or physical properties. Conversely, hydrophilic amino acid residues are characterized by side chain groups that are predominantly polar (e.g., charged or neutrally hydrophilic) in, for example, physiological conditions, where the peptide finds use. Such hydrophilic amino acid residues may be naturally occurring or non-naturally occurring. Hydrophilic amino acid residues may be mimetics of naturally occurring amino acids with predominantly hydrophilic (charged or neutrally polar) side chain groups. Examples of hydrophilic and hydrophobic amino acid residues are shown in Table 1 below. Suitable non-naturally occurring amino acid residues and amino acid mimetics are known in the art. See, e.g., Liang et al. (2013), “An Index for Characterization of Natural and Non-Natural Amino Acids for Peptidomimetics,” PLoS ONE 8(7):e67844.

[0015] Although most amino acid residues can be considered either hydrophobic or hydrophilic, some can behave as either hydrophobic or hydrophilic depending on their context. For example, due to their relatively weak nonpolar character, glycine, proline, serine, and / or cysteine ​​can sometimes function as hydrophilic amino acid residues. Conversely, due to their bulky, slightly hydrophobic side chains, histidine and arginine can sometimes function as hydrophobic amino acid residues.

[0016] [Table 1]

[0017] The term "anti-inflammatory property," as used herein, refers to any property of a polypeptide that reduces or inhibits, or is expected to reduce or inhibit, a pro-inflammatory signal mediated by a protein target and / or reduces or inhibits inflammation in a subject, which can be assessed in silico, in vitro, and / or in vivo. The term "immunomodulatory property," as used herein, refers to any property of a polypeptide that modulates, or is expected to modulate, the expression or secretion of one or more cytokines involved in autoimmune and / or immune responses to infectious pathogens, or that can be assessed in silico, in vitro, and / or in vivo, or by modulating one or more components of a cytokine signaling pathway.

[0018] Selected immunomodulatory peptides of interest The exemplary immunomodulatory peptide sequences described herein are merely examples and are not the only immunomodulatory polypeptides provided herein. Indeed, fragments and variants of the disclosed peptide sequences are also within the scope of the present disclosure.

[0019] The present disclosure provides immunomodulatory polypeptides, sometimes referred to as "RP peptides," that satisfy one or more of the structural formulas set forth below. The present disclosure also provides immunomodulatory polypeptides that share a minimal degree of homology with any of the exemplary RP peptides disclosed herein, or variants thereof, or fragments thereof. Thus, the peptides or polypeptides of the present disclosure are immunomodulatory peptides that satisfy one of the formulas set forth herein or share a minimal degree of homology with any of the exemplary RP peptides disclosed herein.

[0020] "Fragments" of the present invention comprise at least 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, or 23 contiguous amino acid residues (or up to one less than the number of amino acid residues in the subject peptides) of a peptide disclosed herein, and retain at least one immunomodulatory property of the subject peptide. Thus, fragments of the present invention include peptides that lack one, two, three, four, or more amino acids from the N-terminus and / or C-terminus compared to a parent immunomodulatory peptide disclosed herein.

[0021] A "variant" of the present invention is a polypeptide that is substantially similar to a polypeptide disclosed herein and retains at least one immunomodulatory property of the subject polypeptide. A variant may include the deletion (i.e., truncation) of one or more amino acid residues at the N-terminus or C-terminus of a subject polypeptide disclosed herein, the deletion and / or addition of one or more amino acid residues at one or more internal sites in a subject polypeptide disclosed herein, and / or the substitution of one or more amino acid residues (e.g., one, two, three, or even more) at one or more positions in a subject polypeptide disclosed herein. In a subject polypeptide 12 amino acid residues in length or less, a variant polypeptide may include three or fewer (e.g., three, two, one, or none) deleted amino acid residues, whether located internally, at the N-terminus, and / or at the C-terminus.

[0022] Thus, the present invention further provides immunomodulatory polypeptides that are at least 50% identical (i.e., at least 50% sequence identity) (e.g., at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95% or more) to any one of the immunomodulatory polypeptides disclosed in the tables disclosed herein (e.g., Table 3), and still retain at least one immunomodulatory property. Sequence identity is based on a comparison of two peptide sequences or fragments thereof of the same or similar length.

[0023] Thus, in certain embodiments, the present disclosure provides polypeptides comprising an amino acid sequence having 1 to 10 amino acid differences (e.g., 10 or fewer, 9 or fewer, 8 or fewer, 7 or fewer, 6 or fewer, 5 or fewer, 4 or fewer, 3 or fewer, 2 or fewer, or 1 amino acid difference) relative to any one of the polypeptides disclosed herein, while still retaining at least one immunomodulatory property. "Amino acid difference," as used herein, includes amino acid substitutions, amino acid insertions, terminal amino acid additions, amino acid deletions, terminal amino acid truncations, or any combination thereof. Differences between the stripathic region of a homologous immunomodulatory polypeptide and any one of the immunomodulatory polypeptides in Table 3 can include deletions, additions, and / or substitutions of amino acid residues as discussed herein. The substituted amino acid residue may be unrelated to the amino acid residue being substituted (e.g., unrelated in terms of hydrophobicity / hydrophilicity, size, charge, polarity, etc.), or the substituted amino acid residue may constitute a similar, conservative, or highly conservative amino acid substitution. As used herein, "similar," "conservative," and "highly conservative" amino acid substitutions are defined as set forth below in Table 2. The determination of whether an amino acid residue substitution is similar, conservative, or highly conservative is based solely on the side chains of the amino acid residues and not the peptide backbone, which may be modified to increase peptide stability, as discussed below.

[0024] [Table 2]

[0025] Particular immunomodulatory peptides of interest, as well as fragments and variants thereof, that find use in the subject pharmaceutical compositions and methods, are described in more detail below. In certain cases, the subject immunomodulatory peptides have macrophage-modulating activity.

[0026] The "length" of a polypeptide refers to the number of end-to-end amino acid residues comprising the polypeptide, excluding any non-peptide linkers and / or modifications the polypeptide may contain. In some embodiments, the peptide is 5-30 amino acid residues long (e.g., 5-25, 10-20, 5-18, 5-12, 5-10, 6-30, 6-25, 6-20, 6-18, 6-12, 6-10, 7-12, or 7-10 amino acid residues long) and comprises a stripathic region of alternating hydrophilic and hydrophobic modules that adopt an amphipathic conformation under physiological conditions (e.g., as described herein). In some embodiments, the peptide is 5-12 amino acid residues long (e.g., 6, 7, 8, 9, or 10 amino acid residues long) and comprises a stripathic region of alternating hydrophilic and hydrophobic modules that adopt an amphipathic conformation under physiological conditions. In certain cases, the stripathic region of the peptide is 5 to 18 amino acid residues in length (e.g., 6 to 18, 6 to 14, 6 to 12, 7 to 12, or 5, 6, 7, 8, 9, 10, 11, or 12 amino acids in length), and the peptide is optionally further modified (e.g., as described herein). The stripathic region may comprise two or more (e.g., three or more or four or more) hydrophobic modules and one or more (e.g., two or more, three or more, or four or more) hydrophilic modules (e.g., each comprising at least one cationic residue). In some embodiments, a subject immunomodulatory peptide (e.g., as described herein) is a CD206-binding peptide. In some cases, the stripathic region of the peptide has a length of 6 to 12 amino acid residues, such as a length of 7 to 12 amino acid residues. In some cases, the stripathic region of the peptide has a length of 6 to 10 amino acid residues.

[0027] The hydrophobic module can be composed of any suitable residue. In certain cases, the hydrophobic module comprises an amino acid residue selected from phenylalanine, tryptophan, alanine, valine, and glycine. The stripathic region can comprise a total of one, two, or more cationic amino acid residues, such as three or more, four or more, five or more, six or more, or even more. The immunomodulatory peptide can comprise two, three, or more hydrophilic modules composed of any suitable residues. In some cases, the hydrophilic module comprises an amino acid residue selected from lysine, arginine, histidine, aspartic acid, glutamic acid, asparagine, and glutamine.

[0028] In the formulas described herein, J(N) is used to refer to a particular hydrophobic module, where N indicates its position in the linear equation. Similarly, X(N) is used to refer to a particular hydrophilic module, where N indicates its position in the linear equation.

[0029] In the formulas described herein, J (nx) is used to refer to a particular hydrophobic amino acid residue, where n indicates in which module the residue is located and x indicates its position in the module. (nx) is used to refer to a particular hydrophilic amino acid residue, where n indicates in which module the residue is located and x indicates its position in the module.

[0030] In the particular case of the present immunomodulatory peptide, the stripathic region comprises hydrophobic and hydrophilic modules having the following formula: [J1]-[X1]-[J2] (Formula 1)

[0031] In some embodiments of the immunomodulatory peptide, the stripathic region comprises the following formula of hydrophilic and hydrophobic modules: [J1]-[X1]-[J2]-[X2] (Formula 2)

[0032] In some embodiments of the immunomodulatory peptide, the stripathic region comprises the following formula of hydrophilic and hydrophobic modules: [X1]-[J1]-[X2]-[J2] (Formula 3)

[0033] In some embodiments of the immunomodulatory peptides, the stripathic region comprises the following formula of hydrophobic and hydrophilic modules: [J1]-[X1]-[J2]-[X2]-[J3] (Formula 4)

[0034] In certain embodiments, the stripathic region comprises three or more hydrophilic modules and three or more hydrophobic modules and comprises one of the following formulas: [J1]-[X1]-[J2]-[X2]-[J3]-[X3] (Formula 5) [J1]-[X1]-[J2]-[X2]-[J3]-[X3]-[J4] (Formula 6)

[0035] In certain embodiments, the stripathic region comprises three or more hydrophilic modules and three or more hydrophobic modules and comprises the formula: [X1]-[J1]-[X2]-[J2]-[X3]-[J3] (Formula 7)

[0036] In some cases of Formula 1, the stripathic region has a sequence defined by one of the following formulas: [J 1a J 1b ]-[X 1a X 1b ]-[J 2a J 2b ](Formula 1A) [J 2b J 2a ]-[X 1b X 1a ]-[J 1b J 1a ](Formula 1B) During the ceremony, J 1a , J 1b , J 2a , and J. 2bare each independently selected from hydrophobic amino acid residues (e.g., phenylalanine, tryptophan, and valine); X 1a and X 1b are each independently selected from hydrophilic amino acid residues (e.g., lysine or arginine).

[0037] In some instances of Formula 1A, the peptide comprises the sequence FWKRFV(RP837N) (SEQ ID NO: 5), or a fragment or variant thereof (eg, a variant containing one substitution).

[0038] In some embodiments of Formula 2, the stripathic region has a sequence defined by the following formula: [J 1a J 1b ]-[X 1a X 1b ]-[J 2a ]-[X 2a ](Formula 2A) During the ceremony, J 1a , J 1b , and J. 2a are each independently selected from a hydrophobic amino acid residue (e.g., phenylalanine, tryptophan, or valine); X 1a , X 1b , and X 2a are each independently selected from hydrophilic amino acid residues (e.g., lysine or arginine).

[0039] In some instances of Formula 2A, the peptide has the sequence FVRKWR(RP837C 1 ) (SEQ ID NO: 6), or a fragment or variant thereof (eg, a variant containing one substitution).

[0040] In some embodiments of Formula 3, the stripathic region has a sequence defined by the following formula: [X 1a X 1b ]-[J 1a J 1b J 1c J1d ]-[X 2a X 2b ]-[J 2a J 2b ](Formula 3A) During the ceremony, J 1a , J 1b , J 1c , J 1d , J 2a , and J. 2b are each independently selected from a hydrophobic amino acid residue (e.g., leucine, serine, alanine, or phenylalanine); X 1a , X 1b , X 2a , and X 2b are each independently selected from hydrophilic amino acid residues (e.g., glutamic acid, aspartic acid, lysine, asparagine, or arginine).

[0041] In some embodiments of Formula 3A, the stripathic region has a sequence defined by the following formula: EX 1b LSAFX 2a NJ 2a J 2b (SEQ ID NO: 25) During the ceremony, J 2a and J. 2b are each independently selected from alanine and phenylalanine; X 1b and X 2a are each independently selected from lysine and arginine.

[0042] In some instances of Formula 3A, the peptide comprises the sequence EKLSAFRNFF(RP843) (SEQ ID NO: 9), or a fragment or variant thereof (eg, a variant containing one or two substitutions).

[0043] In the particular case of Formula 4, the stripathic region has a sequence defined by one of the following formulas: [J 1a J 1b ]-[X 1aX 1b ]-[J 2a J 2b ]-[X 2a X 2b ]-[J 3a J 3b ](Formula 4A) [J 3a J 3b ]-[X 2a X 2b ]-[J 2b J 2a ]-[X 1b X 1a ]-[J 1b J 1a ](Formula 4B) During the ceremony, J 1a , J 1b , J 2a , J 2b , J 3a , and J. 3b are each independently selected from a hydrophobic amino acid residue (e.g., phenylalanine, tyrosine, isoleucine, or leucine); X 1a , X 1b , X 2a , and X 2b are each independently selected from hydrophilic amino acid residues (e.g., lysine or arginine).

[0044] In some embodiments of Formulas 4A-4B, the striapathic region has a sequence defined by the following formula: LJ 1b KKIIKKJ 3a L (SEQ ID NO: 26) During the ceremony, J. 1b and J. 3a is independently phenylalanine, tyrosine, or leucine (e.g., tyrosine or leucine).

[0045] In some instances of Formulas 4A-4B, the peptide comprises the sequence LYKKIIKKLL (RP846) (SEQ ID NO: 12), or a fragment or variant thereof (eg, a variant containing one or two substitutions).

[0046] In some embodiments of Formula 4, the stripathic region has a sequence defined by the following formula: [J 1a J 1b J 1c ]-[X 1a ]-[J 2a J 2b ]-[X 2a X 2b ]-[J 3a J 3b ](Formula 4C) During the ceremony, J 1a , J 1b , J 1c , J 2a , J 2b , J 3a , and J. 3b are each independently selected from a hydrophobic amino acid residue (e.g., phenylalanine, tyrosine, or proline); X 1a , X 2a , and X 2b are each independently selected from hydrophilic amino acid residues (e.g., aspartic acid, lysine, or arginine).

[0047] In some embodiments of Formula 4C, the stripathic region has a sequence defined by the following formula: FYPDJ 2a J 2b X 2a X 2b J 3a J 3b (SEQ ID NO: 27) During the ceremony, J 2a , J 2b , J 3a , and J. 3b are each independently phenylalanine or tyrosine (e.g., phenylalanine); X 2a and X 2b are each independently lysine or arginine.

[0048] In some instances of Formula 4C, the peptide comprises the sequence FYPDFFKKFF(RP844) (SEQ ID NO: 10), or a fragment or variant thereof (eg, a variant containing one or two substitutions).

[0049] In some embodiments of Formula 4, the stripathic region has a sequence defined by the following formula: [J 1a J 1b ]-[X 1a X 1b ]-[J 2a ]-[X 2a X 2b X 2c ]-[J 3a J 3b ](Formula 4D) During the ceremony, J 1a , J 1b , J 2a , J 3a , and J. 3b are each independently selected from a hydrophobic amino acid residue (e.g., phenylalanine, serine, glycine, or isoleucine); X 1a , X 1b , X 2a , X 2b , and X 2c are each independently selected from hydrophilic amino acid residues (e.g., glutamic acid, aspartic acid, lysine, or arginine).

[0050] In some embodiments of Formula 4D, the stripathic region has a sequence defined by the following formula: J 1a J 1b X 1a X 1b SKEKIG (SEQ ID NO: 28) During the ceremony, J 1a and J. 1b are each independently phenylalanine or tyrosine (e.g., phenylalanine); X 1a and X 1b are each independently lysine or arginine.

[0051] In some instances of Formula 4D, the peptide comprises the sequence FFRKSKEKIG (RP853) (SEQ ID NO: 18), or a fragment or variant thereof (eg, a variant containing one or two substitutions).

[0052] In certain cases, the stripathic region has a sequence defined by the following formula: [J 1a J 1b ]-[X 1a X 1b ]-[J 2a J 2b ]-[X 2a X 2b ]-[J 3a ](Formula 4E) During the ceremony, J 1a , J 1b , J 2a , J 2b , and J. 3a are each independently selected from phenylalanine, alanine, and isoleucine; X 1a , X 1b , X 2a , X 2b , and X 2c are each independently selected from ornithine, lysine, and arginine.

[0053] In certain cases, the stripathic region has a sequence defined by the following formula: [J 1a J 1b ]-[X 1a X 1b ]-[J 2a J 2b ]-[X 2a X 2b ]-[J 3a ]-[X 3a ](Formula 5A) During the ceremony, J 1a , J 1b , J 2a , J 2b , and J. 3aare each independently selected from phenylalanine, tryptophan, alanine, and valine; X 1a , X 1b , X 2a , X 2b , and X 3a are each independently selected from ornithine, lysine, and arginine.

[0054] In some embodiments of Formula 5A, the striapathic region has a sequence defined by the following formula: J 1a J 1b OOJ 2a J 2b OOJ 3a O (SEQ ID NO: 29) During the ceremony, J. 1a , J 1b , J 2a , J 2b , and J. 3a are each independently selected from phenylalanine and alanine (e.g., each J1, J2, and J3 module includes both phenylalanine and alanine).

[0055] In some embodiments of Formula 5A, the striapathic region has a sequence defined by the following formula: FAX 1a X 1b FAX 2a X 2b J 3a FX 3a (SEQ ID NO: 30) In the formula, X 1a , X 1b , X 2a , X 2b , and X 3a are each independently selected from ornithine, lysine, and arginine.

[0056] In some instances of Formula 5A, the peptide comprises the sequence FAOOFAOOFO(RP850) (SEQ ID NO: 19), or a fragment or variant thereof (eg, a variant containing one or two substitutions).

[0057] In some embodiments of Formula 5A, the striapathic region has a sequence defined by the following formula: FWKX 1b FVX 2a KWX 3a (SEQ ID NO: 31) In the formula, X 1b , X 2a , and X 3a are each independently lysine or arginine.

[0058] In some cases of Formula 5A, the peptide comprises the sequence FWKRFVRKWR (RP837) (SEQ ID NO: 4) or FWKKFVKKWK (RP841) (SEQ ID NO: 7), or a fragment or variant thereof (e.g., a variant containing one or two substitutions).

[0059] In some cases, the immunomodulatory peptide of Formula 5A is not FFRKFAKRFK (RP183) (SEQ ID NO: 21) or FFKKFFKKFK (RP185) (SEQ ID NO: 22).

[0060] In certain cases, the stripathic region has a sequence defined by the following formula: [J 1a J 1b ]-[X 1a X 1b ]-[J 2a J 2b ]-[X 2a X 2b ]-[J 3a ]-[X 3a ](Formula 5A) During the ceremony, J 1a , J 1b , J 2a , J 2b , and J. 3a are each independently selected from hydrophobic amino acid residues (e.g., phenylalanine, tryptophan, alanine, valine, and glycine); X 1a , X 1b , X 2a , X 2b , and X 3aare each independently selected from hydrophilic amino acid residues (e.g., lysine, ornithine, arginine, histidine, aspartic acid, glutamic acid, asparagine, or glutamine).

[0061] In some cases of Equation 5A, J 1a , J 1b , J 2a , J 2b , and J. 3a are each independently selected from phenylalanine, tryptophan, alanine, and glycine; 1a , X 1b , X 2a , X 2b , and X 3a are each independently selected from lysine and arginine. In particular cases of Formula 5A, J 1a , J 1b , J 2a , J 2b , and J. 3a are each independently selected from phenylalanine, tryptophan, alanine, and valine; 1a , X 1b , X 2a , X 2b , and X 3a are each independently selected from ornithine, lysine, and arginine (e.g., Lys or Arg). In some instances of Formula 5A, J 1a , J 1b , J 2a , J 2b , and J. 3a are each independently selected from phenylalanine and alanine; 1a , X 1b , X 2a , X 2b , and X 3a are each independently selected from lysine and arginine. In the particular case of Formula 5A, J 1a , J 1b , J 2a , J 2b , and J. 3a are phenylalanines, and X 1a , X 1b , X 2a , X2b , and X 3a are each independently selected from lysine and arginine. 1a , J 1b , J 2a , J 2b , and J. 3a are tryptophan, and X 1a , X 1b , X 2a , X 2b , and X 3a are each independently selected from histidine, lysine, and arginine. In some instances of Formula 5A, J 1a , J 2a , and J. 3a are each independently selected from phenylalanine and tryptophan; 1b is selected from tryptophan and alanine, and J 2b is selected from valine, tryptophan, and alanine, and X 1a , X 1b , X 2a , X 2b , and X 3a are each independently selected from ornithine, lysine, arginine, or histidine.

[0062] In some embodiments of Formula 5A, the striapathic region has a sequence defined by the following formula: WWX 1a HWWHX 2b WX 3a (SEQ ID NO: 32) In the formula, X 1a , X 2b , and X 3a are each independently histidine, lysine, or arginine.

[0063] In some cases of Formula 5B, the peptide comprises the sequence WWHHWWHHWH (RP847) (SEQ ID NO: 13), WWRHWWHRWR (RP848) (SEQ ID NO: 14), or WWKHWWHKWK (RP849) (SEQ ID NO: 15), or a fragment or variant thereof (e.g., a variant containing one or two substitutions).

[0064] In some embodiments of Formula 5, the stripathic region has a sequence defined by the following formula: [J 1a J 1b ]-[X 1a X 1b ]-[J 2a J 2b J 2c ]-[X 2b ]-[J 3a ]-[X 3a ](Formula 5B) During the ceremony, J 1a , J 1b , J 2a , J 2b , and J. 3a are each independently selected from a hydrophobic amino acid residue (e.g., phenylalanine, alanine, threonine, or leucine); X 1a , X 1b , X 2a , X 2b , and X 3a are each independently selected from hydrophilic amino acid residues (e.g., histidine, aspartic acid, lysine, or arginine).

[0065] In some embodiments of Formula 5B, the stripathic region has a sequence defined by the following formula: J 1a J 1b X 1a HJ 2a J 2b THLD (SEQ ID NO: 33) During the ceremony, J 1a , J 1b , J 2a , and J. 2b are each independently selected from phenylalanine and alanine; X 1a is independently selected from lysine and arginine.

[0066] In some instances of Formula 5C, the peptide comprises the sequence FFRHFATHLD (RP845) (SEQ ID NO: 11), or a fragment or variant thereof (eg, a variant containing one or two substitutions).

[0067] In some embodiments of Formula 5, the stripathic region has a sequence defined by the following formula: [J 1a ]-[X 1a ]-[J 2a J 2b J 2c ]-[X 2a ]-[J 3a J 3b ]-[X 3a X 3b ](Formula 5C) During the ceremony, J 1a , J 2a , J 2b , J 2c , J 3a , and J. 3b are each independently selected from a hydrophobic amino acid residue (e.g., phenylalanine, tyrosine, leucine, glycine, or isoleucine); X 1a , X 2a , X 3a , and X 3b are each independently selected from hydrophilic amino acid residues (e.g., glutamine, lysine, or histidine).

[0068] In some embodiments of Formula 5C, the stripathic region has a sequence defined by the following formula: J 1a QJ 2a LGX 2a IIHH (SEQ ID NO: 34) During the ceremony, J 1a and J. 2a are each independently selected from phenylalanine, tyrosine, and leucine; X 2a are lysine and arginine.

[0069] In some instances of Formula 5C, the peptide comprises the sequence FQFLGKIIHH(RP852) (SEQ ID NO: 17), or a fragment or variant thereof (eg, a variant containing one or two substitutions).

[0070] In some embodiments of Formula 6, the stripathic region has a sequence defined by the following formula: [J 1a ]-[X 1a X 1b ]-[J 2a J 2b ]-[X 2a ]-[J 3a ]-[X 3a X 3b ]-[J 4a J 4b ](Formula 6A) During the ceremony, J 1a , J 2a , J 2b , J 3a , J 4a , and J. 4b are each independently selected from hydrophobic amino acid residues (e.g., phenylalanine, tryptophan, alanine, isoleucine, valine, and glycine); X 1a , X 1b , X 2a , X 3a , and X 3b are each independently selected from hydrophilic amino acid residues (e.g., lysine, arginine, histidine, aspartic acid, glutamic acid, asparagine, or glutamine).

[0071] In some embodiments of Formula 6A, the stripathic region has a sequence defined by the following formula: GX 1a X 1b Good job! 2b X 2a GX 3a X 3b Good job! 4b (SEQ ID NO: 35) During the ceremony, J 2b and J. 4bare each independently selected from phenylalanine, tryptophan, alanine, isoleucine, and valine; X 1a , X 1b , X 2a , X 3a , and X 3b are each independently selected from lysine, arginine, histidine, aspartic acid, glutamic acid, asparagine, and glutamine.

[0072] In some embodiments of Formula 6A, the stripathic region has a sequence defined by the following formula: GDX 1b GIX 2a GHX 3b GF (SEQ ID NO: 36) In the formula, X 1b , X 2a , and X 3b are each independently selected from lysine and arginine.

[0073] In some instances of Formula 6A, the peptide comprises the sequence GDRGIKGHRGF(RP842) (SEQ ID NO:8), or a fragment or variant thereof (eg, a variant containing one or two substitutions).

[0074] In some embodiments of Formula 7, the stripathic region has a sequence defined by the following formula: [X 1a X 1b ]-[J 1a ]-[X 2a ]-[J 2a ]-[X 3a ]-[J 3a J 3b J 3c ](Formula 7A) During the ceremony, J 1a , J 2a , J 3a , J 3b , and J. 3c are each independently selected from a hydrophobic amino acid residue (e.g., isoleucine, valine, leucine, serine, or alanine); X 1a , X 1b , X 2a , and X 3a are each independently selected from hydrophilic amino acid residues (e.g., lysine or arginine).

[0075] In some embodiments of Formula 7A, the stripathic region has a sequence defined by the following formula: X 1a X 1b IX 2a VX 3a LSA (SEQ ID NO: 37) In the formula, X 1a , X 1b , X 2a , and X 3a are each independently selected from lysine and arginine.

[0076] In some instances of Formula 7A, the peptide comprises the sequence KKIRVRLSA (RP851) (SEQ ID NO: 16), or a fragment or variant thereof (eg, a variant containing one or two substitutions).

[0077] Multimeric Peptides The present disclosure includes multimers (e.g., dimers) of two or more immunomodulatory peptides (e.g., as described herein) connected via a branched or linear linker. Embodiments of the present disclosure include dimers of any of the subject immunomodulatory polypeptides. Dimers may be homodimers or heterodimers. Any two immunomodulatory polypeptides may be connected via a linker. Any convenient linker may be utilized. Linkers that can be used include, but are not limited to, covalent bonds, peptide linkers (e.g., glycine-containing linkers or Gly and Ser-containing linkers), C1-C12 linkers with terminal amino and / or carboxylic acid groups, or polymer linkers (e.g., PEG or modified PEG). Dimers may include a linker connecting the C-terminus of a first polypeptide to the N-terminus of a second polypeptide. In certain cases, two polypeptides may be linked via their C-terminus. In certain cases, two polypeptides may be linked via their N-terminus.

[0078] The present disclosure further includes any two immunomodulatory polypeptides linked together. The linkage can be formed by a peptide linker such as Gly-Gly-Gly (GGG), Gly-Gly-Gly-Arg (GGGR, SEQ ID NO: 40), Gly-Pro-Gly (GPG), or Gly-Pro-Gly-Arg (GPGR, SEQ ID NO: 41) sequences, linking the C-terminus of a first immunomodulatory polypeptide to the N-terminus of a second immunomodulatory polypeptide. Alternatively, the linkage can be a peptoid linker (e.g., a poly-N-substituted version of any of the aforementioned peptide linkers), a polymer containing g-amino acids (e.g., one corresponding to any of the aforementioned peptide linkers), or a non-peptide chemical linker. The linked immunomodulatory polypeptides can be any of the polypeptides disclosed herein (e.g., Table 3) and can include the same polypeptide linked to form a homodimer or different polypeptides linked to form a heterodimer. Techniques for linking peptides via peptide and non-peptide linkers are well known in the art, and the polypeptide combinations of the present invention are intended to encompass all such linkages.

[0079] Any two stripathic region-containing peptides (e.g., as described herein) can be linked. The two regions of a dimeric peptide can be homodimeric or heterodimeric with respect to each other. A homodimer means that the two peptide regions of a dimeric peptide have the same N-to-C sequence or the reverse C-to-N sequence. The subject immunomodulatory polypeptides described herein can be linked in any convenient configuration to produce multimers. In certain cases, a multimer comprises three or more immunomodulatory polypeptides (e.g., as described herein), and the polypeptides can be arranged in a linear or branched fashion. A linear multimer of an immunomodulatory polypeptide can comprise a head-to-tail arrangement of linked peptides linked via a covalent bond or any linker (e.g., a peptide linker). In some cases, a linear multimer may be referred to as an oligomer, e.g., a polypeptide chain comprising sequence segments of an immunomodulatory polypeptide (e.g., as described herein). Alternatively, the immunomodulatory polypeptides of linear multimers can be linked via a head-to-head (e.g., N-terminus to N-terminus linked) and / or tail-to-tail (e.g., C-terminus to C-terminus linked) configuration. In branched multimers, the immunomodulatory polypeptides can be linked via any convenient branched linker, for example, a group containing three functional groups for binding to amino acid residues such as lysine amino acids. In some cases, the multimer is a dimer.

[0080] In certain cases, the immunomodulatory peptide dimer has the formula: Z 1 -TZ 2 During the ceremony, T is a linker, e.g., a peptide linker; Z 1 is a first polypeptide or region of 3 to 10 (e.g., 4 to 10, 5 to 10, or 3 to 6, or 3, 4, 5, or 6) amino acid residues that is composed of a mixture of hydrophilic and hydrophobic amino acid residues (e.g., as described herein); Z 2is a second polypeptide or region of 3 to 10 (e.g., 4 to 10, 5 to 10, or 3 to 6, or 3, 4, 5, or 6) amino acid residues that is composed of a mixture of hydrophilic and hydrophobic amino acid residues (e.g., as described herein).

[0081] In certain cases of the dimer, the hydrophilic module consists of amino acid residues selected from lysine and arginine, and the hydrophobic module consists of amino acid residues selected from phenylalanine and tryptophan. In certain cases, the first and second polypeptides (Z 1 and Z 2 ) contains four amino acid residues. In certain cases, Z 1 and Z 2 Each of the amino acid residues comprises four amino acid residues, two of which are hydrophilic residues (e.g., as described herein) and the remaining two of which are hydrophobic residues (e.g., as described herein).

[0082] In certain embodiments, the dimer has one of the following formulas: [X1]-[J1]-T-[J1]-[X1] (Formula 8) [J1]-[X1]-T-[X1]-[J1] (Formula 9) [X1]-[J1]-T-[J2]-[X2] (Formula 10) [J1]-[X1]-T-[X2]-[J2] (Formula 11) where T is a linker (e.g., a peptide linker).

[0083] In some cases of Formula 8 and Formula 9, the dimer has a sequence defined by one of the following formulas: [X 1a X 1b ]-[J 1a J 1b ]-T-[J 1b J 1a ]-[X 1b X 1a ](Formula 8A) [J 1a J 1b ]-[X1a X 1b ]-T-[X 1b X 1a ]-[J 1b J 1a ](Formula 9A) During the ceremony, T is a peptide linker (e.g., a polyglycine linker); J 1a and J. 1b are each independently selected from a hydrophobic amino acid residue (e.g., tryptophan or phenylalanine); X 1a and X 1b are each independently selected from hydrophilic amino acid residues (e.g., asparagine or arginine). In the particular cases of Formula 8A and Formula 9A, T is a peptide linker consisting of one, two, or three glycine residues.

[0084] In some embodiments of Formula 9A, the dimer has a sequence defined by the following formula: FW-[X 1a X 1b ]-T-[X 1b X 1a ]-WF (SEQ ID NO: 38) In the formula, X 1a , X 1b are each independently selected from lysine and arginine.

[0085] In some embodiments of Formula 9A, the dimer has a sequence defined by the following formula: [J 1a J 1b ]-KR-T-RK-[J 1b J 1a ] (SEQ ID NO: 39) During the ceremony, J. 1a and J. 1b are each independently selected from tryptophan and phenylalanine.

[0086] In some instances of Formula 7A, the peptide comprises the sequence FWKRGGRKWF(RP837A) (SEQ ID NO: 4), or a fragment or variant thereof (eg, a variant containing one or two substitutions).

[0087] In some cases of Formula 10 and Formula 11, the dimer has a sequence defined by one of the following formulas: [J 1a J 1b ]-[X 1a X 1b ]-T-[X 2a X 2b ]-[J 2a J 2b ](Formula 10A) [X 1a X 1b ]-[J 1a J 1b ]-T-[J 2a J 2b ]-[X 2a X 2b ](Formula 11A) During the ceremony, J 1a , J 1b , J 2a , and J. 2b are each independently selected from a hydrophobic amino acid residue (e.g., tryptophan or phenylalanine); X 1a , X 1b , X 2a , and X 2b are each independently selected from hydrophilic amino acid residues (e.g., asparagine or arginine). In the particular cases of Formula 10A and Formula 11A, T is a peptide linker consisting of one, two, or three glycine residues.

[0088] In certain cases, the first and second polypeptides of the dimer (Z 1 and Z 2 ) has the following formula of hydrophilic and hydrophobic modules: [X1]-[J1]-[X2]-[J2] (Formula 3) or [J1]-[X1]-[J2]-[X2] (Formula 2).

[0089] In certain embodiments, the dimer has one of the following formulas: [X1]-[J1]-[X2]-[J2]-T-[J2]-[X2]-[J1]-[X1] (Formula 12) [J1]-[X1]-[J2]-[X2]-T-[X2]-[J2]-[X1]-[J1] (Formula 13) [X1]-[J1]-[X2]-[J2]-T-[J3]-[X3]-[J4]-[X4] (Formula 14) [J1]-[X1]-[J2]-[X2]-T-[X3]-[J3]-[X4]-[J4] (Formula 15) where T is a peptide linker.

[0090] In some cases of Formula 12 and Formula 13, the dimer has one of the following formulas: [X 1a ]-[J 1a ]-[X 2a ]-[J 2a ]-T-[J 2a ]-[X 2a ]-[J 1a ]-[X 1a ](Formula 12A) [J 1a ]-[X 1a ]-[J 2a ]-[X 2a ]-T-[X 2a ]-[J 2a ]-[X 1a ]-[J 1a ](Formula 13A) During the ceremony, T is a peptide linker (e.g., a polyglycine linker); J 1a and J. 2a are each independently selected from phenylalanine and tryptophan; X 1a and X 2a are each independently selected from lysine and arginine.

[0091] In some instances of Formula 12A, the peptide comprises the sequence RWKFGGFKWR (RP832C) (SEQ ID NO: 1), or a fragment or variant thereof (eg, a variant containing one or two substitutions).

[0092] In some instances of Formula 13A, the peptide comprises the sequence FKWRGGRWKF(RP837C) (SEQ ID NO: 3), or a fragment or variant thereof (e.g., a variant containing one or two substitutions). In certain embodiments, the immunomodulatory peptide comprises a tail region.

[0093] In some instances of Formula 14 and Formula 15, the dimer has one of the following formulas: [X 1a ]-[J 1a ]-[X 2a ]-[J 2a ]-T-[J 3a ]-[X 3a ]-[J 4a ]-[X 4a ](Formula 14A) [J 1a ]-[X 1a ]-[J 2a ]-[X 2a ]-T-[X 3a ]-[J 3a ]-[X 4a ]-[J 4a ](Formula 15A) During the ceremony, T is a peptide linker (e.g., a polyglycine linker); J 1a , J 2a , J 3a , and J. 4a are each independently selected from phenylalanine and tryptophan; X 1a , X 2a , X 3a , and X 4a are each independently selected from lysine and arginine.

[0094] Immunomodulatory peptides of interest include, but are not limited to, any one of the polypeptides in Table 3, a fragment thereof (e.g., as described herein), or a variant thereof (e.g., as described herein).

[0095] [Table 3]

[0096] In certain embodiments, a subject immunomodulatory polypeptide is a) a sequence selected from the peptide sequences of Table 3; b) a sequence having at least 75% sequence identity (e.g., at least 80%, at least 85%, at least 90%, or at least 95% sequence identity) with a sequence defined in a), and c) a sequence having one or two amino acid substitutions compared to the sequence defined in a), wherein the one or two amino acid substitutions are amino acid substitutions according to Table 2 (e.g., similar, conservative, or highly conservative amino acid substitutions). The sequence includes a sequence selected from:

[0097] In certain cases, the sequence described in a) is RP832C. In certain cases, the sequence described in a) is RP837. In certain cases, the sequence described in a) is RP837C. In certain cases, the sequence described in a) is RP837A. In certain cases, the sequence described in a) is RP837N. In certain cases, the sequence described in a) is RP837C. 1In particular cases, the sequence described in a) is RP841. In particular cases, the sequence described in a) is RP842. In particular cases, the sequence described in a) is RP843. In particular cases, the sequence described in a) is RP844. In particular cases, the sequence described in a) is RP845. In particular cases, the sequence described in a) is RP846. In particular cases, the sequence described in a) is RP847. In particular cases, the sequence described in a) is RP848. In particular cases, the sequence described in a) is RP849. In particular cases, the sequence described in a) is RP850. In particular cases, the sequence described in a) is RP851. In particular cases, the sequence described in a) is RP852. In particular cases, the sequence described in a) is RP853.

[0098] In certain cases, the sequence described in b) has a sequence that has at least 80% sequence identity with the sequence defined in a). In certain cases, the sequence described in b) has a sequence that has at least 85% sequence identity with the sequence defined in a). In certain cases, the sequence described in b) has a sequence that has at least 90% sequence identity with the sequence defined in a). In certain cases, the sequence described in b) has a sequence that has at least 95% sequence identity with the sequence defined in a).

[0099] In certain embodiments, the sequence shown in c) has one or two amino acid substitutions compared to the sequence defined in a), wherein the one or two amino acid substitutions are similar amino acid substitutions according to Table 2. In certain embodiments, the sequence shown in c) has one or two amino acid substitutions compared to the sequence defined in a), wherein the one or two amino acid substitutions are conservative amino acid substitutions according to Table 2. In certain embodiments, the sequence shown in c) has one or two amino acid substitutions compared to the sequence defined in a), wherein the one or two amino acid substitutions are highly conservative amino acid substitutions according to Table 2. Any of the variations of immunomodulatory peptides described herein may be applied to the parent peptides of Table 3.

[0100] Excluded Polypeptides The compositions of the present disclosure optionally exclude polypeptides described in U.S. Patent Application Nos. 2012 / 0270770 and 2003 / 0109452, and U.S. Patent No. 6,559,281, the disclosures of which are incorporated herein by reference in their entireties. Accordingly, one or more polypeptides described in such publications and / or the use of such polypeptides may be excluded from the scope of the compositions and / or methods of the present disclosure. Additionally, any of the polypeptides disclosed in Tables 3-9 of WO 2016 / 061133 by Jaynes et al. (the disclosures of which tables are incorporated herein by reference) may be optionally excluded from the compositions and / or methods of using such compounds disclosed herein. In some cases, any of the polypeptides disclosed in Table 4 below may be optionally excluded from the compositions and / or methods of using such compounds disclosed herein.

[0101] In some cases, the immunomodulatory peptide of the formulae described herein is not a polypeptide of Table 4.

[0102] [Table 4]

[0103] Modified Polypeptides Embodiments of the present disclosure include modification of any of the immunomodulatory polypeptides of the present disclosure by chemical or genetic means. Examples of such modifications include constructing partial or complete peptides with unnatural and / or natural amino acids in the L- or D-form. For example, any of the peptides disclosed herein and any variants thereof can be produced in an all-D form. Furthermore, polypeptides of the present disclosure can be modified to contain carbohydrate or lipid moieties, such as sugars or fatty acids, covalently attached to the side chains of amino acids or to the N- or C-terminus. In addition, polypeptides of the present disclosure can be modified to enhance solubility and / or half-life upon administration. For example, polyethylene glycol (PEG) and related polymers have been used to enhance the solubility and half-life of protein therapeutics in the blood. Thus, polypeptides of the present disclosure can be modified with PEG polymers, etc. Polypeptides of the present disclosure can also be modified to contain sulfur, phosphorus, halogens, metals, etc. Amino acid mimetics can be used to generate polypeptides of the present disclosure (e.g., having a structure based on a structural algorithm or a structure similar to any of the immunomodulatory polypeptides disclosed herein). In certain embodiments, polypeptides of the present disclosure that include amino acid mimetics have enhanced properties, such as resistance to degradation. For example, a polypeptide of the present disclosure can include one or more (e.g., all) peptoid monomers.

[0104] The immunomodulatory polypeptide can be linked to another molecule via a biodegradable linkage, such as a disulfide bond. The disulfide bond can be mediated by the sulfhydryl group of a cysteine ​​residue found in the immunomodulatory polypeptide and a sulfhydryl group in another molecule. The cysteine ​​residue can be located, for example, at either the C-terminus or N-terminus of the immunomodulatory polypeptide. This type of disulfide linkage can be used to conveniently link the polypeptide of the present disclosure to various types of useful molecules. For example, the linkage can be with another immunomodulatory polypeptide (optionally containing a C-terminus or N-terminus cysteine ​​residue), a fluorescent label (e.g., Dylight350), a chemotherapeutic agent (e.g., a taxol derivative formed by adding a sulfhydryl group to an appropriate site on the taxol ring structure and then oxidizing it with a cysteine-containing peptide of the present disclosure), etc.

[0105] The linked immunomodulatory polypeptides (e.g., homodimers or heterodimers) can bind to a target molecule (e.g., a target protein such as a pro-inflammatory signaling protein) with a binding energy greater than the binding energy of either monomeric polypeptide alone. Thus, for example, the binding energy of the linked immunomodulatory polypeptides to an NF-kB class II protein (e.g., RelB) can be at least -700 kcal / mol, and in certain embodiments, at least -750, -800, -900, -1000, -1100, -1200, -1250, -1300, -1350, -1400, -1425, -1450, -1475, -1500, -1525, -1550, -1575, -1600 kcal / mol, or more. The energy of binding can be determined using methods well known in the art (eg, using the ClusPro™ algorithm), for example, in silico, in vitro, or in vivo.

[0106] In some cases where a modified peptide is covalently linked to a molecule of interest, the resulting compound may be referred to as a peptide conjugate. Any convenient molecule of interest may be conjugated to the subject immunomodulatory peptides. The molecule of interest may be peptidic or non-peptidic, naturally occurring, or synthetic. Molecules of interest suitable for use in combination with the subject immunomodulatory peptides include, but are not limited to, moieties that confer desirable drug-like properties, such as protein domains, polypeptides, peptide tags, specific binding moieties (e.g., antibodies or antibody fragments), polymer moieties such as polyethylene glycol (PEG), carbohydrates, dextran, or polyacrylate, linkers, half-life extending moieties, labels, and solid supports. In some cases, the molecule of interest may confer enhanced and / or modified properties and functions to the resulting modified peptide, including, but not limited to, increased water solubility, ease of chemical synthesis, cost, bioconjugation sites, stability, pI, aggregation, reduced nonspecific binding, and / or specific binding to a second target protein, as described herein.

[0107] In some embodiments where any one of the peptide sequences described herein is set forth, the peptide sequence may be extended to include one or more additional residues at the N-terminus and / or C-terminus of the sequence, e.g., two or more, three or more, four or more, five or more, six or more, or even more additional residues. Any convenient residue can be included at the N-terminus and / or C-terminus of the peptide to provide a desirable property or group, such as increased solubility via a water-soluble group, a linkage for dimerization or multimerization, a linkage for connecting to a label or specific binding moiety, etc.

[0108] In some instances, the subject modified peptides are described by the following formula: BLM wherein B is an immunomodulatory peptide (e.g., as described herein), L is an optional linking group, M is a molecule of interest, and L is attached to B at any convenient position (e.g., at the N-terminus, C-terminus, or via the side chain of a residue not involved in binding to the target).

[0109] The modified peptide may include one or more molecules of interest. In some cases, the molecule of interest is covalently attached via the alpha-amino group of the N-terminal residue or to the alpha-carboxylic acid group of the C-terminal residue.

[0110] Molecules of interest may include polypeptides or protein domains, including, but not limited to, gD tags, c-Myc epitopes, FLAG tags, His tags, fluorescent proteins (e.g., GFP), beta-galactosidase proteins, GST, albumin, immunoglobulins, antibodies, Fc domains or similar antibody-like fragments, leucine zipper motifs, coiled-coil domains, hydrophobic regions, hydrophilic regions, polypeptides containing free thiols that form intermolecular disulfide bonds between two or more multimerization domains, "bulge into cavity" domains, beta-lactoglobulin, or fragments thereof.

[0111] The subject molecules may comprise a half-life extending moiety. The term "half-life extending moiety" refers to a pharmaceutically acceptable moiety, domain, or "vehicle" covalently attached or conjugated to a subject compound that prevents or reduces chemical modifications that reduce proteolysis or other activity of the subject compound in vivo, increases half-life or other pharmacokinetic properties (e.g., absorption rate), reduces toxicity, improves solubility, increases the biological activity and / or target selectivity of the subject compound for a target of interest, increases manufacturability, and / or reduces the immunogenicity of the subject compound, compared to the unconjugated form of the subject compound.

[0112] In certain embodiments, the half-life extending moiety is a polypeptide that binds to a serum protein, such as an immunoglobulin (e.g., IgG) or serum albumin (e.g., human serum albumin (HSA)). Polyethylene glycol is one example of a useful half-life extending moiety. Exemplary half-life extending moieties include polyalkylene glycol moieties (e.g., PEG), serum albumin or fragments thereof, transferrin receptor or transferrin-binding portion thereof, and moieties comprising a binding site for a polypeptide that enhances in vivo half-life, copolymers of ethylene glycol, copolymers of propylene glycol, carboxymethylcellulose, polyvinylpyrrolidone, poly-1,3-dioxolane, poly-1,3,6-trioxane, ethylene / maleic anhydride copolymers, polyamino acids (e.g., polylysine), dextran n-vinylpyrrolidone, poly-vinylpyrrolidone, propylene glycol homopolymers, propylene oxide poly(propylene oxide), and the like. polymers, ethylene oxide polymers, polyoxyethylated polyols, polyvinyl alcohols, linear or branched glycosylated chains, polysialic acid, polyacetals, lipids, long chain fatty acids, long chain hydrophilic aliphatic groups, immunoglobulin Fc domains (see, e.g., U.S. Pat. No. 6,660,843), albumin (e.g., human serum albumin, see, e.g., U.S. Pat. Nos. 6,926,898 and US 2005 / 0054051, 6,887,470), transthyretin (TTR, see, e.g., U.S. Pat. Nos. 2003 / 0195154, 2003 / 0191056), or thyroxine-binding globulin (TBG).

[0113] In certain embodiments, the half-life extending moiety is a lipid. In certain embodiments, the half-life extending moiety is a fatty acid. Any convenient lipid and fatty acid can be used in the subject modified compounds. See, for example, Chae et al., "The fatty acid conjugated exendin-4 analogs for type 2 antidiabetic therapeutics," J. Control Release. 2010 May 21; 144(1):10-6.

[0114] In certain embodiments, the immunomodulatory peptide is modified to include a specific binding moiety. A specific binding moiety is a moiety that can specifically bind to a second moiety that is complementary thereto. In some cases, the specific binding moiety has a K of 100 nM or less (e.g., 30 nM or less, 10 nM or less, 3 nM or less, 1 nM or less, 300 pM or less, or 100 pM or less). D (measured by) at least 10 -7 It binds to a complementary second moiety with an affinity of M. Complementary binding moiety pairs of specific binding moieties include, but are not limited to, ligands or activators / promoters and receptors, antibodies and antigens, complementary polynucleotides, complementary protein homo- or heterodimers, aptamers and small molecules, and polyhistidine tags and nickel. Specific binding pairs can also include analogs, derivatives, and fragments of the original specific binding member. For example, an antibody directed against a protein antigen can also recognize peptide fragments, chemically synthesized and labeled proteins, derivatized proteins, etc., as long as the epitope is present. Protein domains of interest that find use as specific binding moieties include, but are not limited to, Fc domains or similar antibody-like fragments, leucine zipper motifs, coiled-coil domains, hydrophobic regions, hydrophilic regions, polypeptides containing free thiols that form intermolecular disulfide bonds between two or more multimerization domains, or "bulge into cavity" domains (see, e.g., WO 94 / 10308; U.S. Pat. No. 5,731,168; Lovejoy et al. (1993), Science 259: 1288-1293; Harbury et al. (1993), Science 262: 1401-05; Harbury et al. (1994), Nature 371: 80-83; Hakansson et al. (1999), Structure 7: 255-64).

[0115] In certain embodiments, the peptide is a linked specific binding moiety that specifically binds to a target protein. The linked specific binding moiety can be an antibody, antibody fragment, receptor activator, or aptamer. The linked specific binding moiety can specifically bind to any convenient target protein, for example, a target protein that is desirable to target in conjunction with the subject therapeutic methods. Target proteins of interest include, but are not limited to, PDGF (e.g., PDGF-B), VEGF-B, VEGF-C, VEGF-D, EGF, EGFR, Her2, PD-1, PD-L1, OX-40, and LAG3. In certain embodiments, the linked specific binding moiety is a receptor activator or ligand, for example, a protein ligand associated with an inflammatory pathway, such as interleukin-13 (IL-13), or a molecule that activates a member of the Toll-like receptor (TLR) family, for example, TLR3. In certain cases, the linked specific binding moiety (e.g., a protein, antibody, or antibody fragment) can be further linked to an additional active agent (e.g., a chemotherapeutic agent described herein).

[0116] Immunomodulatory polypeptides (e.g., as described herein) can be conjugated to additional active agents to provide immunomodulatory polypeptide conjugates. Once the subject peptides have been generated and / or produced and selected according to the teachings herein, they can be linked, fused, conjugated (e.g., covalently or noncovalently), or otherwise associated with pharmaceutically active or diagnostic moieties or biocompatible modifiers. The term "peptide conjugate" refers to any biologically active or detectable molecule or drug associated with the disclosed immunomodulatory peptide compounds, regardless of the method of association. In this regard, it is understood that such conjugates can include, in addition to the disclosed immunomodulatory peptides, peptides, polypeptides, proteins, prodrugs that are metabolized in vivo to active agents, polymers, nucleic acid molecules, small molecules, binding agents, mimetics, synthetic drugs, inorganic molecules, organic molecules, and radioisotopes. Furthermore, as noted above, the selected conjugates may be covalently or non-covalently associated with or linked to the subject peptides and may exhibit various stoichiometric molar ratios depending, at least in part, on the method used to effect the conjugation.

[0117] In certain cases, the molecule of interest is a second active agent, e.g., an active agent or drug, that finds use in combination with a target of interest in the subject therapeutic methods. In certain cases, the molecule of interest is a small molecule, a chemotherapeutic agent, an antibody, an antibody fragment, a bispecific antibody, an aptamer, or an L-protein. In some embodiments, the peptide is modified to include a moiety useful as a pharmaceutical (e.g., a protein, a nucleic acid, a small organic molecule, etc.). Exemplary pharmaceutical proteins include, for example, cytokines, antibodies, chemokines, growth factors, interleukins, cell surface proteins, extracellular domains, cell surface receptors, cytotoxins, etc. Exemplary small molecule pharmaceuticals include small molecule toxins or therapeutic agents. Any convenient therapeutic or diagnostic agent (e.g., as described herein) can be conjugated to the immunomodulatory peptide. A variety of therapeutic agents, including, but not limited to, anticancer agents, antiproliferative agents, cytotoxic agents, and chemotherapeutic agents, are described below in the section entitled "Combination Therapy," any one of which may be adapted for use in the subject peptide conjugates.

[0118] In certain embodiments, the modified peptides can be conjugated to bispecific antibodies, for example, engineered bispecific monoclonal antibodies that can simultaneously bind to two different types of antigens of interest.

[0119] In certain embodiments, the modified peptide may include a cell-penetrating peptide (e.g., tat). Cell-penetrating peptides can facilitate cellular uptake of molecules. Any convenient tag polypeptide and its respective antibody may be used. Examples include poly-histidine (poly-his) or poly-histidine-glycine (poly-his-gly) tags; influenza HA tag polypeptide and its antibody 12CA5 [Field et al., Mol. Cell. Biol. 8:2159-2165 (1988)]; c-myc tag and its corresponding antibodies 8F9, 3C7, 6E10, G4, B7, and 9E10 [Evan et al., Molecular and Cellular Biology, 5:3610-3616 (1985)]; and herpes simplex virus glycoprotein D (gD) tag and its antibody [Paborsky et al., Protein Engineering, 3(6):547-553 (1990)]. Other tag polypeptides include Flag peptide [Hopp et al., BioTechnology 6:1204-1210 (1988)], KT3 epitope peptide [Martin et al., Science 255:192-194 (1992)], tubulin epitope peptide [Skinner et al., J. Biol. Chem. 266:15163-15166 (1991)], and T7 gene 10 protein peptide tag [Lutz-Freyermuth et al., Proc. Natl. Acad. Sci. USA 87:6393-6397 (1990)].

[0120] Those skilled in the art will appreciate that several different reactions are available for attaching or associating therapeutic or diagnostic moieties and / or linkers to the subject immunomodulatory peptides. In certain embodiments, this can be achieved by reacting amino acid residues of, for example, the peptides described herein, including the amino terminus, the C-terminal carboxylic acid, the amine group of lysine, the free carboxylic acid groups of glutamic acid and aspartic acid, the sulfhydryl group of cysteine, and various portions of aromatic amino acids. One method of covalent attachment is the carbodiimide reaction, which links the carboxy (or amino) group of a compound to the amino (or carboxy) group of the subject peptides. In addition, bifunctional agents such as dialdehydes or imidoesters have been used to link the amino group of the subject peptides to the amino group of an antibody molecule. Maleimide-thiol conjugation chemistry, e.g., click chemistry between an azide group and an alkynyl group, can also be used to attach drugs to immunomodulatory peptides. The Schiff base reaction can also be used to attach drugs to peptides. This method can involve periodate oxidation of a drug containing a glycol or hydroxy group to form an aldehyde, which is then reacted with the binder. Conjugation occurs via the formation of a Schiff base with the amino group of the binder. Isothiocyanates and azlactones can also be used as coupling agents to covalently attach drugs to binders.

[0121] It is understood that several variations or types of linkers can be used to associate the disclosed immunomodulatory peptides with pharmaceutically active or diagnostic moieties or biocompatible modifiers. In some embodiments, the linker is cleavable under intracellular conditions, so that cleavage of the linker releases the drug unit from the antibody in the intracellular environment. In certain embodiments, the linker unit is not cleavable. Bivalent linker reagents useful for linking two or more functional or biologically active moieties, such as peptides, nucleic acids, drugs, toxins, antibodies, haptens, and reporter groups, are known, and methods for the resulting conjugates have been described (Hermanson, GT (1996) Bioconjugate Techniques; Academic Press: New York, p234-242).

[0122] composition The composition of the present disclosure comprises an immunomodulatory polypeptide that satisfies one of the structural formulas described herein.For example, the immunomodulatory polypeptide may have a stripathic region that has a sequence according to any one of the formulas disclosed herein.Typically, the immunomodulatory polypeptide contained in the composition of the present disclosure is a synthetic polypeptide (e.g., produced by chemical synthesis and / or recombinantly produced).

[0123] The compositions of the present disclosure may contain a single immunomodulatory polypeptide or a combination thereof. The compositions may be substantially free of proteins and other polypeptides that do not meet the structural algorithms disclosed herein. As used herein, the term "substantially free of proteins and other polypeptides" means that less than 5% of the protein content of the composition is made up of proteins and other polypeptides that are not immunomodulatory polypeptides of the present disclosure. A composition that is substantially free of non-immunomodulatory polypeptides of the present disclosure may have 4%, 3%, 2%, 1%, 0.5%, 0.1%, 0.05%, 0.01%, or less of proteins or other polypeptides that do not meet the structural algorithms disclosed herein. Thus, the compositions may be substantially free of blood proteins such as serum albumin, globulins, fibrinogen, and clotting factors. Alternatively, the compositions may be substantially free of globulins, fibrinogen, and clotting factors, but may contain purified or recombinantly produced serum albumin.

[0124] In certain embodiments, the compositions of the present disclosure contain immunomodulatory polypeptides that are not naturally found in humans or other mammals or animals. However, the compositions of the present disclosure may also contain immunomodulatory polypeptides that are naturally found in humans or other mammals or animals, provided that the compositions are substantially free of biological molecules (such as non-immunomodulatory polypeptides, nucleic acids, lipids, carbohydrates, and metabolites) that associate with or co-purify with the immunomodulatory polypeptides in vivo. As used herein, the term "substantially free of biological molecules" means that less than 5% of the dry weight of the composition is made up of biological molecules that are not immunomodulatory polypeptides. A composition that is substantially free of such biological molecules may have 4%, 3%, 2%, 1%, 0.5%, 0.1%, 0.05%, 0.01%, or less of biological molecules that are not immunomodulatory polypeptides. Thus, for example, the compositions may be substantially free of biological molecules that are abundant in blood, such as the proteins, fatty acids, cholesterol, non-protein clotting factors, and metabolites discussed above. In addition, the composition may be substantially free of cells, including red blood cells, white blood cells, and platelets, and cell fragments.

[0125] Compositions of the present disclosure may comprise at least 1 mg (e.g., at least 5, 10, 20, 30, 40, 50, 75, 100, 150, 200, 250, 300, 400, 500, 600, 700, 800, 900, 1000 mg, or more) of an immunomodulatory polypeptide. Thus, for example, the composition may contain from about 1 mg to about 1000 mg (e.g., from about 5 mg to about 900 mg, from about 5 mg to about 800 mg, from about 5 mg to about 700 mg, from about 5 mg to about 600 mg, from about 10 mg to about 500 mg, from about 10 mg to about 400 mg, from about 10 mg to about 300 mg, from about 10 mg to about 250 mg, from about 10 mg to about 200 mg, from about 10 mg to about 150 mg, from about 10 mg to about 100 mg, from about 50 mg to about 500 mg, from about 50 mg to about 400 mg, from about 50 mg to about 300 mg, from about 50 mg to about 250 mg, from about 50 mg to about 200 mg, The immunomodulatory polypeptide may comprise an amount equal to about 50 mg to about 150 mg, about 50 mg to about 100 mg, about 75 mg to about 500 mg, about 75 mg to about 400 mg, about 75 mg to about 300 mg, about 75 mg to about 250 mg, about 75 mg to about 200 mg, about 75 mg to about 150 mg, about 75 mg to about 100 mg, about 100 mg to about 500 mg, about 100 mg to about 400 mg, about 100 mg to about 300 mg, about 100 mg to about 250 mg, about 100 mg to about 200 mg, or any other range containing two of the above-mentioned endpoints.

[0126] Compositions of the present disclosure can include solutions containing at least 1 mg / ml (e.g., at least 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100 mg / ml or more) of an immunomodulatory polypeptide. Thus, for example, compositions can include solutions containing from about 1 mg / ml to about 1000 mg / ml (e.g., from about 5 mg / ml to about 900 mg / ml, from about 5 mg / ml to about 800 mg / ml, from about 5 mg / ml to about 700 mg / ml, from about 5 mg / ml to about 600 mg / ml, from about 5 mg / ml to about 500 mg / ml, from about 10 mg / ml to about 500 mg / ml, from about 10 mg / ml to about 400 mg / ml, or from about 10 mg / ml to about 1000 mg / ml). ~300mg / ml, approximately 10mg / ml~250mg / mL, approximately 10mg / mL~200mg / mL, approximately 10mg / mL~150mg / mL, approximately 10mg / mL~100mg / mL, Approximately 50 mg / mL to approximately 500 mg / mL, approximately 50 mg / mL to approximately 400 mg / mL, approximately 50 mg / mL to approximately 300 mg / mL, approximately 50 mg / mL to approximately 250 mg / mL, approximately 50 mg / mL to approximately 20 0mg / mL, about 50mg / mL to about 150mg / mL, about 50mg / mL to about 100mg / mL, about 75mg / mL to about 500mg / ml, about 75mg / ml to about 400mg / ml, about 75m g / ml~about 300mg / ml, about 75mg / ml~about 250mg / ml, about 75mg / ml~about 200mg / ml, about 75mg / ml~about 150mg / ml, about 75mg / ml~about 100mg The immunomodulatory polypeptide may comprise a solution having a concentration of about 100 mg / ml, about 100 mg / ml to about 500 mg / ml, about 100 mg / ml to about 400 mg / ml, about 100 mg / ml to about 300 mg / ml, about 100 mg / ml to about 250 mg / ml, about 100 mg / ml to about 200 mg / ml, about 10 mg / ml to about 150 mg / ml, or any other range containing two of the aforementioned endpoints.

[0127] The compositions of the present disclosure include pharmaceutical compositions. Such pharmaceutical compositions may include one or more immunomodulatory polypeptides and a pharmaceutically acceptable carrier. The pharmaceutical composition may further include proteins and / or chemotherapeutic agents other than the immunomodulatory polypeptides of the present disclosure. The other protein may be a therapeutic agent, such as a therapeutic antibody. The therapeutic protein or antibody may have immunomodulatory properties or other properties that are enhanced by or are enhanced by the immunomodulatory polypeptides of the present disclosure. Alternatively, the other protein may be a carrier protein, such as serum albumin (e.g., HSA). Serum albumin (e.g., HSA, BSA, etc.) may be purified or recombinantly produced. Mixing the immunomodulatory polypeptide(s) in the pharmaceutical composition with serum albumin can effectively "load" the immunomodulatory polypeptide onto the serum albumin, allowing for greater delivery of the immunomodulatory polypeptide to the site of inflammation. The chemotherapeutic agent may be, for example, an anti-cancer chemotherapeutic agent. Such chemotherapeutic agents include, but are not limited to, gemcitabine, docetaxel, bleomycin, erlotinib, gefitinib, lapatinib, imatinib, dasatinib, nilotinib, bosutinib, crizotinib, ceritinib, trametinib, bevacizumab, sunitinib, sorafenib, trastuzumab, ado-trastuzumab emtansine, rituximab, ipilimumab, rapamycin, temsirolimus, everolimus, methotrexate, doxorubicin, Abraxane, Forfirinox, cisplatin, carboplatin, 5-fluorouracil, Teysumo, paclitaxel, prednisone, levothyroxine, and pemetrexed.

[0128] In some instances of the subject pharmaceutical compositions, the compositions comprise an immunomodulatory polypeptide that is a CD206-binding peptide (e.g., as described herein) and a chemotherapeutic agent. In some embodiments, the immunomodulatory polypeptide that finds use in a combination composition is a peptide in Table 3. In particular instances, an immunomodulatory peptide (e.g., a peptide in Table 3) is combined with a chemotherapeutic agent. In particular instances of the pharmaceutical composition, the chemotherapeutic agent is gemcitabine. In some instances of the pharmaceutical composition, the chemotherapeutic agent is docetaxel. In some instances of the pharmaceutical composition, the chemotherapeutic agent is Abraxane.

[0129] In some instances of the subject pharmaceutical compositions, the compositions comprise an immunomodulatory polypeptide that is a CD206-binding peptide (e.g., as described herein) conjugated to a second additional agent (e.g., as described herein). In some instances, the additional agent is a chemotherapeutic agent. In some embodiments, the immunomodulatory polypeptide that finds use in the subject peptide conjugates is a peptide in Table 3. In particular instances, an immunomodulatory peptide (e.g., a peptide in Table 3) is conjugated to a chemotherapeutic agent. In particular instances of the subject peptide conjugates, the chemotherapeutic agent is gemcitabine. In some instances of the subject peptide conjugates, the chemotherapeutic agent is docetaxel. In some instances of the subject peptide conjugates, the chemotherapeutic agent is Abraxane. In some instances of the subject peptide conjugates, the chemotherapeutic agent is paclitaxel.

[0130] In some cases, the subject pharmaceutical compositions find use in the treatment of cancer, e.g., ovarian cancer, comprise an immunomodulatory polypeptide in combination with a vaccination therapy, e.g., a dendritic cell (DC) vaccinating agent that promotes Th1 / Th17 immunity. In some of the pharmaceutical compositions, the immunomodulatory polypeptide is an adjuvant in combination with a Th17-inducing vaccination agent.

[0131] The pharmaceutical compositions of the present invention can be formulated for oral administration, parenteral administration, inhalation administration, topical administration, mucosal administration, and the like. In some embodiments, administration is via a route selected from oral, intravenous, intraperitoneal, inhalation, intranasal, intraprostatic, and intratumoral. The present invention is not limited by the route of administration. Compositions formulated for oral delivery can include, for example, an enteric coating to ensure that the peptide contained therein reaches the intestinal tract and beyond. Enteric formulations, such as gastroresistant capsules for oral administration and suppositories for rectal or vaginal administration, also form part of the present disclosure. Compositions formulated for topical delivery can be, for example, suspended in a gel or cream, coated onto microneedles, or injected into a bandage or topical patch to extend the duration of action of the peptide contained therein. Any inhalable formulation capable of providing an aerosolized form containing the subject peptide for delivery to a patient via the pulmonary route can be used in conjunction with the present disclosure. In some cases, the subject compositions are administered, for example, by injectable dermal, subcutaneous, and / or intratumoral injection into a nodular tumor.

[0132] In some embodiments, the compositions are administered mucosally (e.g., using standard techniques; see, e.g., Remington: The Science and Practice of Pharmacy, Mack Publishing Company, Easton, Pa., 19th edition, 1995 (e.g., for mucosal delivery techniques, including intranasal, pulmonary, vaginal, and rectal techniques), and European Publication No. 517,565 and Illum et al., J. Controlled Rel., 1994, 29:133-141 (e.g., for intranasal administration techniques)). In some cases, the compositions of the invention can be administered dermally or transdermally using standard techniques. Methods of intranasal vaccination include administering a subject composition in droplet or spray form to the nasopharynx of the subject to be treated. In some embodiments, sprayed or aerosolized compositions are provided.

[0133] Also provided are liposomal pharmaceutical compositions containing the subject immunomodulatory peptides. Any convenient nanocarriers and liposomes can be adapted for use in preparing liposomal formulations of the subject peptides, such as the nanocarriers and liposomes described by Arias in "Liposomes in drug delivery: a patent review," Expert Opinion on Therapeutic Patents, 23, 2013, issue 11, pp. 1399-1414, and by Torchilin in "Multifunctional nanocarriers," Advanced Drug Delivery Reviews, Volume 58, Issue 14, December 1, 2006, Pages 1532-1555.

[0134] Nanoparticle formulations or compositions containing the subject immunomodulatory peptides are also provided. The nanoparticle formulations or compositions can increase the water solubility of the peptides of interest, achieving protected, sustained, and targeted delivery of the peptides in therapeutic applications (e.g., as described herein). In some cases, the formulations are polymer-based nanoparticle formulations. Nanoparticle formulations of interest include albumin nanoparticles, e.g., human serum albumin-containing nanoparticle formulations. In some cases, desolvation techniques can be used to prepare albumin nanoparticles. Particle size, peptide drug release, encapsulation efficiency, and peptide drug-polymer interactions can be determined and selected using any convenient in vitro method. For example, cell culture studies of in vivo pharmacokinetics in rats can be used to evaluate the biological properties of the desired formulation.

[0135] In some instances, nanoparticle formulation compositions containing the subject immunomodulatory peptides are comprised of iron oxide nanoparticles (IONPs). IONPs find use in a variety of biomedical applications. In some cases, IONP formulations can exhibit high uptake within macrophages and / or target cancer cells. IONPs with desired cytotoxicity, in vivo distribution, and / or clearance can be selected for use in combination with the subject immunomodulatory peptides. A variety of well-characterized IONPs with different sizes and coatings can be utilized in the subject compositions and formulations. In some cases, polyethyleneamine (PEI)-coated IONPs or PEGylated IONPs are utilized. IONPs can enhance the cytotoxicity of the subject formulations through multiple mechanisms, such as ROS generation and apoptosis.

[0136] Also provided are kits comprising an immunomodulatory polypeptide that is a CD206-binding peptide (e.g., as described herein) and an additional agent (e.g., a chemotherapeutic or immunotherapeutic agent) for use in treating cancer. The kit may include a dose of the immunomodulatory peptide in an amount effective to inhibit the growth of cancer cells in a subject. The kit may also include a dose of an additional agent, such as a chemotherapeutic or immunotherapeutic agent (e.g., as described herein), in an amount effective to inhibit the growth of cancer cells in a subject. In some cases, the kit includes an insert with instructions for administering the immunomodulatory peptide and / or the additional agent (e.g., a chemotherapeutic or immunotherapeutic agent). In some cases, the set of instructions for combination therapy may recommend (i) a lower dose of the immunomodulatory peptide when used in combination with a chemotherapeutic agent, (ii) a lower dose of the additional agent (e.g., a chemotherapeutic or immunotherapeutic agent) when used in combination with the immunomodulatory peptide, and / or (iii) a dosing regimen of one or both agents that differs from the normally recommended dosing regimen.

[0137] method The present disclosure provides methods of modulating macrophage activity using immunomodulatory peptides (e.g., as described herein). In some methods, the modulated macrophage activity is macrophage polarization. The method can include contacting macrophages with a CD206-binding agent that is a peptide of the present disclosure to modulate macrophage activity. In some cases, modulating macrophage activity refers to inhibiting macrophage activity. The methods of the present invention can provide a reduction in macrophage viability, which can be determined using any convenient method.

[0138] In certain embodiments, immunomodulatory polypeptides of the present disclosure can bind to human CD206 with an affinity of at least -650 kcal / mol, and in certain embodiments, at least -700, -750, -800, -850, -900, -925, -950, -975, -1000, -1025, -1050 kcal / mol or greater. The required binding affinity may correspond to a binding affinity that can be detected in vitro or in vivo. Alternatively, the required binding affinity may correspond to a binding affinity that can be detected in silico, for example, using the ClusPro™ algorithm.

[0139] Macrophages targeted using the subject methods can be M2 macrophages or tumor-associated macrophages (TAMs). Targeted macrophages can be in vitro or in vivo.

[0140] In certain embodiments, the peptides of the present disclosure bind to two or more targets (e.g., pro-inflammatory targets). In some embodiments, the variant polypeptides bind to three, four, five, or more pro-inflammatory targets. For example, the variant polypeptides can bind to any combination of the targets disclosed herein (e.g., NF-kB class II protein and human serum albumin (HSA)), as discussed below. Such binding can be based on in silico, in vitro, or in vivo data.

[0141] Exemplary RP peptides of interest can interact with various signaling molecules associated with inflammation, including the NF-kB class II subunit RelB, TGFβ, Notch1, Wnt8R, TRAIL, IL6R, IL10R, EGFR, and CDK6, as well as other membrane-associated signaling molecules, including CD206, CD47, and SIRP-α, the translational modifying protein transglutaminase 2 (TGM2), and the histone-modifying enzyme histone methyltransferase (HMT). In certain cases, the subject peptides are CD206-binding peptides. Folding these protein targets into their normal three-dimensional conformations often generates amphipathic clefts with high affinity for the immunomodulatory peptides described herein.

[0142] Further details of the target signaling molecules to which the subject immunomodulatory peptides specifically bind are described in WO2016 / 061133 by Jaynes et al., the disclosure of which is incorporated herein in its entirety.

[0143] An immunomodulatory polypeptide that finds use in the subject methods can be based on its ability to bind to the mannose-binding site on CD206 and / or to interfere with or block the binding of SIRP-mannose to CD206. For example, the immunomodulatory polypeptide can bind to at least one amino acid residue of CD206 selected from the group consisting of Glu-725, Tyr-729, Glu-733, Asn-747, and Asp-748, or the equivalent amino acid residue(s) in a CD206 protein of another species. Alternatively, the immunomodulatory polypeptide can be linked to at least one amino acid residue in human CD206 selected from the group consisting of Phe-708, Thr-709, Trp-710, Pro-714, Glu-719, Asn-720, Trp-721, Ala-722, Glu-725, Tyr-729, Glu-733, Asn-747, Asp-748, Ser-1691, Cys-1693, Phe-1694, and Phe-1703, or to the equivalent amino acid residue(s) in a CD206 protein of another species. In certain embodiments, the immunomodulatory polypeptide can be linked to at least one amino acid residue in CD206 selected from the group consisting of Phe-708, Trp-710, Trp-721, Glu-725, Tyr-729, Glu-733, or to the equivalent amino acid residue(s) in a CD206 protein of another species.

[0144] In certain cases, the immunomodulatory polypeptide binds to the fibronectin (FBN) domain of CD206 and / or interferes with or blocks collagen binding to CD206. In some cases, the immunomodulatory polypeptide can specifically bind to the fibronectin (FBN) domain of CD206. In some cases, the subject immunomodulatory polypeptide binds to the C-type carbohydrate recognition domain (CRD) of CD206 and regulates (e.g., activates) the activity of CD206. In some cases, the subject immunomodulatory polypeptide binds to the C-type carbohydrate recognition domain (CRD) of CD206 and regulates (e.g., interferes with, blocks, or inhibits) the activity of CD206. In certain cases, the CRD domain to which the subject immunomodulatory polypeptide specifically binds and regulates the activity of CD206 is the CRD4 or 5 domain.

[0145] In certain embodiments, the immunomodulatory polypeptide binds to two or more targets (e.g., pro-inflammatory targets). In some embodiments, the immunomodulatory polypeptide binds to three, four, five, or more pro-inflammatory targets. For example, the immunomodulatory polypeptide may bind to any combination of the targets disclosed herein. Such binding may be based on in silico, in vitro, or in vivo data. Thus, the immunomodulatory polypeptide may bind to two or more NF-kB class II subunits (e.g., RelB and at least one other NF-kB class II subunit, such as RelA, cRel, NF-kB1, or NF-kB2). Alternatively (or in addition), the immunomodulatory polypeptide can bind to an NF-kB class II subunit (e.g., RelB) and at least one other signaling molecule (e.g., at least one signaling molecule selected from the group consisting of TGFβ, Notch1, Wnt8R, TRAIL, IL6R, IL10R, EGFR, CDK6, CD206, CD47, SIRP-α, HMT, and TGM2). For example, the immunomodulatory polypeptide can bind to an NF-kB class II subunit (e.g., RelB) and at least one signaling molecule selected from the group consisting of TGFβ, Notch1, Wnt8R, TRAIL, IL6R, IL10R, EGFR, and CDK6. Alternatively, the immunomodulatory polypeptide can bind to an NF-kB class II subunit (e.g., RelB) and at least one signaling molecule selected from the group consisting of CD206, CD47, SIRP-α, and TGM2. In another alternative, the immunomodulatory polypeptide can bind to an NF-kB class II subunit (e.g., RelB) and HMT. In another alternative, the immunomodulatory polypeptide can bind to at least one signaling molecule selected from the group consisting of TGFβ, Notch1, Wnt8R, TRAIL, IL6R, IL10R, EGFR, and CDK6, and at least one signaling molecule selected from the group consisting of CD206, CD47, SIRP-α, and TGM2.In another alternative, the immunomodulatory polypeptide binds to at least one signaling molecule selected from the group consisting of TGFβ, Notch1, Wnt8R, TRAIL, IL6R, IL10R, EGFR, and CDK6, and can also bind to an HMT. In yet other embodiments, the immunomodulatory polypeptide can also bind to at least one signaling molecule selected from the group consisting of NF-kB class II subunit (e.g., RelB), TGFβ, Notch1, Wnt8R, TRAIL, IL6R, IL10R, EGFR, and CDK6, at least one signaling molecule selected from the group consisting of CD206, CD47, SIRP-α, and TGM2, and an HMT. In certain embodiments, the immunomodulatory polypeptide also binds to two or more pro-inflammatory targets and serum albumin (e.g., human serum albumin).

[0146] The immunomodulatory polypeptides of the present disclosure provide a powerful tool for reducing inflammation and / or treating conditions associated with excessive inflammation (whether acute or chronic). As used herein, the terms "treat," "treating," and similar words shall mean stabilizing, reducing the symptoms of, preventing the occurrence of, or curing a medical condition.

[0147] Thus, the present disclosure provides methods for reducing the expression level and / or activity of at least one (e.g., two, three, four, five, or more) pro-inflammatory cytokine(s) at a site of inflammation in a subject. The method includes administering to the subject an immunomodulatory polypeptide of the present disclosure (or, e.g., a pharmaceutical composition comprising an immunomodulatory polypeptide). The pro-inflammatory cytokine may be selected from the group consisting of NF-kB, TNFα, IL-1, IL-6, IL-8, IL-12, IL-17, IL-23, MCP-1, MMP-1, and MMP-9. The reduction may be at least 10% (e.g., 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, or more) reduction in cytokine expression or activity.

[0148] The present disclosure also provides methods for inhibiting an increase in the expression level and / or activity of at least one (e.g., two, three, four, five, or more) pro-inflammatory cytokine(s) at a potential site of inflammation in a subject. The method includes administering to the subject an immunomodulatory polypeptide of the present disclosure (or, e.g., a pharmaceutical composition including an immunomodulatory polypeptide). The pro-inflammatory cytokine may be selected from the group consisting of NF-kB, TNFα, IL-1, IL-6, IL-8, IL-12, IL-17, IL-23, MCP-1, MMP-1, and MMP-9. The method can inhibit such an increase by limiting the increase in cytokine expression and / or activity to 20% or less (e.g., 15%, 12.5%, 10%, 7.5%, 5%, 4%, 3%, 2%, 1%, or less).

[0149] It is understood that modulation of the level and / or activity of pro-inflammatory cytokine(s) at a site of inflammation in a subject can, in some cases, provide downstream modulation of the activity of targeted immune cells (e.g., effector T cells, regulatory T cells (Tregs), natural killer cells (NK cells), B cells, etc.) and control of the targeted immune or inflammatory response.

[0150] The present disclosure also provides methods for treating or preventing conditions associated with chronic inflammation, such as irritable bowel disease, ulcerative colitis, colitis, Crohn's disease, idiopathic pulmonary fibrosis, asthma, keratitis, arthritis, osteoarthritis, rheumatoid arthritis, autoimmune diseases, feline or human immunodeficiency virus (FIV or HIV) infection, cancer, age-related inflammation and / or stem cell dysfunction (e.g., age-related increases in Nlrp3 expression, age-related increases in SOCS3 in muscle stem cells, etc.), graft-versus-host disease (GVHD), keloids, scleroderma, obesity, diabetes, diabetic wounds, other chronic wounds, atherosclerosis, multiple sclerosis, Parkinson's disease, Alzheimer's disease, macular degeneration, gout, gastric ulcers, gastritis, mucositis, toxoplasmosis, and chronic viral or microbial infections (e.g., chronic bacterial or protozoal infections, etc.). The method includes administering an immunomodulatory polypeptide of the present disclosure (or, for example, a pharmaceutical composition comprising an immunomodulatory polypeptide) to a subject suffering from or likely to develop the condition.

[0151] The present disclosure also provides a method for treating or preventing fibrosis. Fibrosis can be, for example, pulmonary fibrosis, skin fibrosis, liver fibrosis, kidney fibrosis, or fibrosis caused by ionizing radiation. The method includes administering an immunomodulatory polypeptide of the present disclosure (or, for example, a pharmaceutical composition comprising an immunomodulatory polypeptide) to a subject suffering from or likely to develop fibrosis.

[0152] The present disclosure also provides a method for treating cancer. The cancer may be colon cancer, breast cancer, leukemia, lymphoma, ovarian cancer, prostate cancer, liver cancer, lung cancer, testicular cancer, cervical cancer, bladder cancer, endometrial cancer, renal cancer, melanoma, thyroid cancer, or brain cancer, or ophthalmic cancer. The method comprises administering an immunomodulatory polypeptide of the present disclosure (or, for example, a pharmaceutical composition comprising an immunomodulatory polypeptide) to a subject suffering from cancer. The presently disclosed subject matter also provides a method for treating solid tumor cancer in a subject. In some embodiments, the method comprises administering to the subject a therapeutically effective amount of a compound disclosed herein.

[0153] For any of the foregoing methods, the subject may be an animal such as a livestock animal (e.g., a horse, cow, pig, goat, sheep, rabbit, chicken, turkey, duck, etc.), a pet (e.g., a dog, cat, rabbit, hamster, gerbil, bird, fish, etc.), a laboratory animal (e.g., a mouse, rat, monkey, chimpanzee, owl, fish, etc.), a zoo animal (e.g., a gorilla, orangutan, chimpanzee, monkey, elephant, camel, zebra, wild boar, lion, tiger, giraffe, bear, bird, etc.), a wild animal (e.g., a deer, wolf, mountain lion, bird, etc.), or a human.

[0154] In combination with any of the aforementioned methods, the immunomodulatory polypeptide(s) can be administered at a dose and frequency that depends on the type of animal, the size of the animal, and the condition being treated. Typically, the immunomodulatory polypeptide is administered in a dose of about 1 mg to about 1000 mg (e.g., about 5 mg to about 900 mg, about 5 mg to about 800 mg, about 5 mg to about 700 mg, about 5 mg to about 600 mg, about 10 mg to about 500 mg, about 10 mg to about 400 mg, about 10 mg to about 300 mg, about 10 mg to about 250 mg, about 10 mg to about 200 mg, about 10 mg to about 150 mg, about 10 mg to about 100 mg, about 50 mg to about 500 mg, about 50 mg to about 400 mg, about 50 mg to about 300 mg, about 50 mg to about 250 mg, about 50 mg to about 200 mg, about 5 ... The dose is administered once daily (or every other day or once weekly) in an amount ranging from 0 mg to about 150 mg, about 50 mg to about 100 mg, about 75 mg to about 500 mg, about 75 mg to about 400 mg, about 75 mg to about 300 mg, about 75 mg to about 250 mg, about 75 mg to about 200 mg, about 75 mg to about 150 mg, about 75 mg to about 100 mg, about 100 mg to about 500 mg, about 100 mg to about 400 mg, about 100 mg to about 300 mg, about 100 mg to about 250 mg, about 100 mg to about 200 mg, or any other range containing two of the above endpoints. The daily dose may be administered once during the day or divided into smaller doses taken at multiple times during the day. In humans (and other similarly sized mammals), a dose of 5 mg / kg can be administered every other day. The immunomodulatory polypeptide can be administered for a period of time (e.g., 2-3 weeks), at intervals (e.g., administer the polypeptide for 2-3 weeks, wait 2-3 weeks, then repeat the cycle), or until such time as pro-inflammatory cytokine levels are reduced or stabilized, chronic inflammatory conditions or fibrosis are resolved, or the cancer goes into remission.

[0155] Administration of an immunomodulatory polypeptide (or pharmaceutical composition comprising such a polypeptide) in combination with any of the aforementioned methods can be performed intravenously, intraperitoneally, parenterally, orthotopic, subcutaneously, topically, via inhalation, nasally, orally, sublingually, intraocularly, by means of an implantable depot, using nanoparticle-based delivery systems, microneedle patches, microparticles, beads, osmotic or mechanical pumps, and / or other mechanical means.

[0156] In conjunction with any of the foregoing methods, an immunomodulatory polypeptide (e.g., as described herein) (or a pharmaceutical composition comprising such a polypeptide) can be administered in combination with another drug designed to reduce or prevent inflammation, treat or prevent chronic inflammation or fibrosis, or treat cancer. In each case, the immunomodulatory polypeptide can be administered before, simultaneously with, or after the administration of the other drug. For the treatment of cancer, the immunomodulatory polypeptide(s) can be administered in combination with an additional therapeutic agent (e.g., a chemotherapeutic agent or an immunomodulatory agent) selected from the group consisting of taxanes, nucleoside analogs, steroids, anthracyclines, thyroid hormone replacement drugs, thymidylate-targeting drugs, chimeric antigen receptor / T-cell therapy, chimeric antigen receptor / NK cell therapy, apoptosis regulator inhibitors (e.g., B-cell CLL / lymphoma 2 (BCL-2) BCL-2-like 1 (BCL-XL) inhibitors), CARP-1 / CCAR1 (cell division cycle and apoptosis regulator 1) inhibitors, colony-stimulating factor-1 receptor (CSF1R) inhibitors, CD47 inhibitors, cancer vaccines (e.g., Th17-inducing dendritic cell vaccines), and other cell therapies. Specific chemotherapeutic agents include, for example, gemcitabine, docetaxel, bleomycin, erlotinib, gefitinib, lapatinib, imatinib, dasatinib, nilotinib, bosutinib, crizotinib, ceritinib, trametinib, bevacizumab, sunitinib, sorafenib, trastuzumab, ado-trastuzumab emtansine, rituximab, ipilimumab, rapamycin, temsirolimus, everolimus, methotrexate, doxorubicin, Abraxane, Forfirinox, cisplatin, carboplatin, 5-fluorouracil, Teismo, paclitaxel, prednisone, levothyroxine, pemetrexed, navitoclax, and ABT-199.

[0157] In some embodiments, immunomodulatory polypeptides that find use in combination therapy are peptides with macrophage-modulating activity (e.g., as described herein). In particular cases, immunomodulatory polypeptides are CD206-binding peptides (e.g., as described herein). In some embodiments, immunomodulatory polypeptides that find use in combination therapy are peptides in Table 3. In particular cases, immunomodulatory peptides (e.g., peptides in Table 3) can be administered in combination with a chemotherapeutic agent to treat cancer. In particular cases, the chemotherapeutic agent is gemcitabine. In some cases, the chemotherapeutic agent is docetaxel. In some cases, the chemotherapeutic agent is Abraxane.

[0158] For the treatment of cancer (e.g., melanoma, non-small cell lung cancer, or lymphoma such as Hodgkin's lymphoma), the immunomodulatory polypeptide(s) can be administered in combination with an immunotherapeutic agent. An immunotherapeutic agent is any convenient agent that finds use in treating disease by inducing, enhancing, or suppressing an immune response. In some cases, the immunotherapeutic agent is an immune checkpoint inhibitor. Any convenient checkpoint inhibitor, including but not limited to, cytotoxic T-lymphocyte-associated antigen 4 (CTLA-4) inhibitors, programmed death 1 (PD-1) inhibitors, and PD-L1 inhibitors, can be utilized in combination with the subject peptides. Exemplary checkpoint inhibitors of interest include, but are not limited to, ipilimumab, pembrolizumab, and nivolumab. In certain embodiments, for the treatment of cancer and / or inflammatory diseases, the immunomodulatory polypeptide(s) can be administered in combination with a colony-stimulating factor-1 receptor (CSF1R) inhibitor. CSF1R inhibitors of interest include, but are not limited to, emactuzumab.

[0159] Any convenient cancer vaccine therapy and drug can be used in combination with the subject immunomodulatory polypeptide composition and method.For the treatment of cancer, for example, ovarian cancer, immunomodulatory polypeptide(s) can be administered in combination with vaccine therapy, for example, dendritic cell (DC) vaccination agent that promotes Th1 / Th17 immunity.Th17 cell infiltration is correlated with a significant prolongation of overall survival in ovarian cancer patients.In some cases, immunomodulatory polypeptides find use as adjuvant therapy in combination with Th17-inducing vaccination.

[0160] Also of interest are agents that are CD47 inhibitors, including but not limited to CARP-1 / CCAR1 (cell cycle and apoptosis regulator 1) inhibitors, including but not limited to those described in Rishi et al., Journal of Biomedical Nanotechnology, Volume 11, Number 9, September 2015, pp. 1608-1627 (20), and anti-CD47 antibody agents such as Hu5F9-G4.

[0161] In certain cases, the combination provides an enhanced effect compared to either component alone, and in some cases, the combination provides a supra-additive or synergistic effect compared to the combined or additive effect of the components. Various combinations of the subject polypeptides and chemotherapeutic agents can be used, either sequentially or simultaneously. For multiple doses, the two agents can be directly alternating, or two or more doses of one agent can be alternated, for example, with a single dose of the other agent. Co-administration of both agents can also be alternating, or the dosages of the individual agents can be differently spaced. In some cases, the time between doses can be from about 1-6 hours, to about 6-12 hours, to about 12-24 hours, to about 1-2 days, to about 1-2 weeks, or longer, after the start of treatment.

[0162] In some embodiments, the method is a method of reducing cancer cell proliferation, comprising contacting cells with an effective amount of a subject immunomodulatory polypeptide (e.g., as described herein). The method can be performed in combination with a chemotherapeutic agent (e.g., as described herein). The cancer cells can be in vitro or in vivo. In certain cases, the method comprises contacting cells with an immunomodulatory peptide (e.g., a peptide in Table 3) and contacting the cells with a chemotherapeutic agent. Any advantageous cancer cells can be targeted. In certain cases, the chemotherapeutic agent is gemcitabine. In some cases, the chemotherapeutic agent is docetaxel. In some cases, the chemotherapeutic agent is Abraxane.

[0163] Alternatively, for methods of treating cancer, the immunomodulatory polypeptide(s) (or pharmaceutical compositions comprising such polypeptides) can be administered in combination with radiation therapy. Again, the immunomodulatory polypeptide(s) can be administered before or after the administration of radiation therapy.

[0164] Any of the aforementioned methods of the present disclosure further include a step of evaluating the effectiveness of the therapeutic treatment. Because the immunomodulatory polypeptides of the present disclosure have the demonstrable ability to reduce tissue inflammation and suppress the excessive production of inflammatory mediators such as IL-1, IL-6, IL-12, and TNFα in both tissue and serum (data not shown), the effectiveness of the therapeutic treatment can be evaluated by measuring the levels of such cytokines (e.g., in serum) to determine whether the levels responded appropriately to treatment. Depending on the cytokine levels, the dosage of the immunomodulatory polypeptide(s) can be adjusted up or down as necessary.

[0165] definition It is to be understood that the present invention is not limited to the particular embodiments described herein, which may, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting, since the scope of the present invention will be limited only by the appended claims.

[0166] Where a range of values ​​is provided, unless the context clearly dictates otherwise, it is understood that each intervening value, to the tenth of the unit of the lower limit, between the upper and lower limits of that range, and any other stated or intervening value in that stated range, is encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included in the smaller ranges and, subject to any specifically excluded limit in the stated range, are also encompassed within the invention. Where the stated range includes one or both of the limits, ranges excluding either or both of those included limits are also included in the invention.

[0167] Certain ranges are presented herein with numerical values ​​preceded by the term "about." The term "about" is used herein to provide literal support for the exact number it precedes, as well as a number that is near or approximately the number preceded by the term. In determining whether a number is near or approximately a specifically recited number, the near or approximately unrecited number may be a number that, in the context in which it is presented, provides a substantial equivalent to the specifically recited number.

[0168] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although any methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the present invention, representative exemplary methods and materials are described herein.

[0169] All publications and patents cited herein are incorporated by reference to disclose and describe the methods and / or materials in connection with which the publications are cited, as if each individual publication or patent was specifically and individually indicated to be incorporated by reference. The citation of any publication is for its disclosure prior to the filing date and should not be construed as an admission that the present invention is not entitled to antedate such publication by virtue of prior invention. Further, the publication dates provided may be different from the actual publication dates, which may need to be independently confirmed.

[0170] It should be noted that, as used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. It should be further noted that the claims may be drafted to exclude any and all optional elements. Accordingly, this statement is intended to serve as a predicate for the use of exclusive terminology such as "solely," "only," and the like, or the use of a "negative" limitation in connection with the recitation of claim elements.

[0171] As will be apparent to one of ordinary skill in the art upon reading this disclosure, each of the individual embodiments described and illustrated herein has individual components and features which may be readily separated from or combined with the features of any of the other several embodiments without departing from the scope or spirit of the invention. Any recited method can be carried out in the order of events recited or in any other order which is logically possible. [Example]

[0172] Methods and materials of interest that find use in preparing and evaluating the subject immunomodulatory peptides include those disclosed in the experimental section of WO2016 / 061133 by Jaynes et al., the disclosure of which is incorporated herein in its entirety.

[0173] The following examples are put forth so as to provide those of ordinary skill in the art with a complete disclosure and description of how to make and use the present invention, and are not intended to limit the scope of what the inventors regard as their invention, nor are they intended to represent that the following experiments are all or the only experiments performed.

[0174] Example 1: Inhibition of tumor growth Polypeptides of the present disclosure are also tested for their effect on tumor growth in a mouse model of non-metastatic breast cancer. MCF-7 human non-metastatic breast cancer cells are cultured in normal growth medium at 37°C and 5% CO2. Cells are harvested at 80%-90% confluence. Immune-compromised athymic nude mice (J:NU) are divided into two groups (9 mice per group). All mice receive approximately 4.5 x 10 cells stained with VIVO Tracker 680 and suspended in 200 μl of PBS / Matrigel mixture. 6 MCF-7 cells are injected subcutaneously onto the dorsal surface of the treated animal using a 22-gauge needle fitted with a 500 μl syringe.

[0175] Animals are treated with the designated vehicle and peptide. Peptide-treated animals are treated with the subject polypeptide. Freshly prepared peptides are dissolved in sterile saline at a concentration of 100 μM and used to treat animals in the peptide group. Vehicle-treated animals are injected with saline buffer alone. All treatments are injected into the tumor mass twice a week for 5 weeks using a 27.5-gauge needle attached to a 1 ml syringe. Animal weight and tumor volume are measured three times a week, and after fluorescent labeling, VIVO Tracker 680 and IVIS imaging are performed.

[0176] Figure 2 shows that peptides RP832C and RP185 reduce tumor volume in a mouse tumor inhibition model. The data demonstrate that polypeptides of the present disclosure suppress tumor growth in vivo.

[0177] Example 2: Administration of peptides in combination with chemotherapy Given the significant role of inflammation in tumor development and metastasis, and the known association between M2 macrophage activity and tumor progression, it was anticipated that administration of peptides of the present disclosure (e.g., selected peptides in Table 3) may positively impact the outcome of cancer treatment.

[0178] To test this theory, approximately 5 × 10 mice were injected under the left upper nipple of a cohort of immune-compromised ("nude") mice. 6 Human triple-negative breast cancer cells (MDA-MB-231) are injected into the mice. Following this administration, one cohort receives vehicle only, and two of the cohorts receive the chemotherapy drug gemcitabine at a dose of 40 mg / kg body weight every four days. One of these cohorts also receives the test peptide at a daily dose of 5 mg / kg body weight, and the fourth cohort receives the peptide alone at a daily dose of 5 mg / kg body weight. Starting on day 32 of the study, the concentration of RP-182 is increased to 20 mg / kg body weight in the gemcitabine + RP-182 cohort. Tumor volume is assessed at various time points after initial cell administration. After 50 days, the mice are sacrificed.

[0179] In the second experiment, C42B prostate cancer cell xenografts were introduced into four cohorts of mice, and tumors were grown approximately 100 m before treatment. 3 One cohort is treated with vehicle alone, a second cohort with docetaxel at 2.5 mg / kg body weight once weekly, a third cohort with the test peptide administered subcutaneously at 10 mg / kg body weight once daily, and a fourth cohort with both docetaxel at 2.5 mg / kg once weekly and the test peptide at 10 mg / kg once daily. Tumor volume is assessed at various time points after initial cell administration, and mice are sacrificed 27 days later.

[0180] The peptides of the present disclosure (e.g., selected peptides in Table 3) are expected to produce synergistic effects when administered with chemotherapeutic agents, including gemcitabine and docetaxel, as well as checkpoint inhibitor therapy and other immunotherapies. Specifically, the peptides of the present disclosure may be particularly useful when used in combination with the recently developed CAR-T (chimeric antigen receptor / T cell) therapy. While destroying tumor cells, such therapy also results in a very high systemic load of dead cell material, overstimulating the immune system and resulting in a "cytokine storm" that can be fatal to the patient.

[0181] Example 3: Investigation of selected peptides

[0182] [Table 5]

[0183] Example 4: Selective effects of targeted RP peptides on scleroderma macrophage viability Peripheral blood-derived macrophages were cultured from healthy volunteers and scleroderma patients and cultured in MCS-F for 7 days before qPCR for arginase 1 (M2 marker) or IFNg (M1 marker). They were then treated with RP peptides ranging in dose from 0 to 100 μM. After 48 h, the medium was replaced and the cells were re-treated with peptides. After 96 h, cell viability was assayed by PrestoBlue assay. (Figure 4A) Macrophages from healthy controls with a low arginase:IFNg ratio of 3.6 were resistant to the effects of the targeted RP peptides (RP182, RP185, RP832C, RP837) on viability. (Figure 4B) In scleroderma (SSc) patients with an elevated arginase:IFNg ratio of 8.8, the targeted RP peptides (RP182, RP185, RP832C, RP837), even at 0.01 μM, significantly reduced viability at 96 h.

[0184] Example 5: Bleomycin pulmonary fibrosis rescue Intratracheal instillation (IT) of bleomycin was used as a model of pulmonary fibrosis. Rescue of bleomycin-induced pulmonary fibrosis in mice by the subject peptides was studied. Experimental parameters: Four groups (n = 8) of male C57BL6 mice were studied over a 6-8 week period. 2.5 U / kg bleomycin was administered IT in a single bolus. 72 hours later, 1 mg / kg of the peptide of interest (RP182, RP185, RP832, RP837) was administered IT every other day. In this bleomycin challenge experiment, fibrosis was measured by Ashcroft score after trichrome staining. Collagen score was measured quantitatively after hydroxyproline staining. Figure 1 shows a graph of these results.

[0185] The subject peptide reduced fibrosis and collagen deposition. Hematoxylin and eosin (H / E) and Mason's trichrome staining of lung tissue sections was performed. The alveoli of the vehicle group appeared surrounded by fibrotic tissue with increased collagen deposition, unlike those of the treated and naive lung groups. Body weight change: The body weight of the RP peptide-treated group showed no significant change compared to that of the vehicle-treated group. Ashcroft score analysis: Unlike the peptide-treated group, which showed well-organized lung structure and was assigned a lower score, lung specimens from the vehicle group were assigned a score close to 6 because they showed distorted lung structure with increased fibrosis and collagen deposition.

[0186] Fibrosis and collagen deposition level assessment: There was a significant decrease in fibrosis and collagen deposition in the treatment group compared to the vehicle group, as measured by ImageJ software. Lung weight change: The vehicle group had higher lung weight compared to the peptide-treated group due to the decrease in fibrosis and collagen content in the peptide-treated group. IHC staining of TGFβ1 and αSMA: Lung tissues treated with peptides showed a significant decrease in fibrosis-related markers, TGFβ1 and αSMA.

[0187] These results demonstrate that the exemplary peptides provided a reduction in bleomycin-induced pulmonary fibrosis in a mouse model of pulmonary fibrosis.

[0188] Example 6: Collaboration of RP peptides of interest with PD-1 checkpoint inhibitors in CT26 xenografts Figure 2 shows the results of a study of the effect of peptides of interest, with or without anti-PD1 antibodies, on tumor volume in a mouse tumor inhibition model. Peptides RP832C and RP185 were administered at 10 mg / kg once daily. Anti-PD1 antibodies were administered intraperitoneally at 200 μg twice weekly. The results provided in the Examples demonstrate the efficacy of the peptides of the present invention.

[0189] Regardless of the scope of the appended claims, the following notes are provided to illustrate aspects of the present disclosure.

[0190] An immunomodulatory peptide having a length of 1.5 to 30 (e.g., 6 to 30 or 6 to 18) amino acid residues, An immunomodulatory peptide comprising a stripathic region of alternating hydrophilic and hydrophobic modules (e.g., having a length of 6-12 or 6-10 amino acid residues) described by one of Formulas 1-7, which comprises two or more (e.g., three or more or four or more) hydrophobic modules and one or more (e.g., two or more or three or more) hydrophilic modules, each comprising at least one cationic residue, and which adopts an amphipathic conformation under physiological conditions, and which specifically binds to CD206. 2. The striated region is that of formula 5, and has the formula: [J 1a J 1b ]-[X 1a X 1b ]-[J 2a J 2b ]-[X 2a X 2b ]-[J 3a ]-[X 3a (Formula 5A) wherein J 1a , J 1b , J 2a , J 2b, and J. 3a are each independently selected from hydrophobic amino acid residues (e.g., phenylalanine, tryptophan, alanine, valine, and glycine); and X 1a , X 1b , X 2a , X 2b , and X 3a are each independently selected from a hydrophilic amino acid residue (e.g., lysine, arginine, histidine, aspartic acid, glutamic acid, asparagine, or glutamine). 3.J 1a , J 1b , J 2a , J 2b , and J. 3a are each independently selected from phenylalanine, tryptophan, alanine, and glycine; 1a , X 1b , X 2a , X 2b , and X 3a are each independently selected from lysine and arginine. 4.J 1a , J 1b , J 2a , J 2b , and J. 3a are each independently selected from phenylalanine, tryptophan, alanine, and valine; 1a , X 1b , X 2a , X 2b , and X 3a are each independently selected from ornithine, lysine, and arginine. 5.Formula:FAX 1a X 1b FAX 2a X 2b J 3a FX 3a (SEQ ID NO: 30), wherein X 1a , X 1b , X 2a , X 2b , and X 3aare each independently selected from ornithine, lysine, and arginine.

[0191] 6. The immunomodulatory peptide of appendix 5 having the sequence FAOOFAOOFO (SEQ ID NO: 19) (RP850). 7. Formula:FWKX 1b FVX 2a KWX 3a (SEQ ID NO: 31), wherein X 1b , X 2a , and X 3a The immunomodulatory peptide according to any one of appendix 2 to 4, wherein each of is independently lysine or arginine. 8. The immunomodulatory peptide of appendix 7, having a sequence selected from FWKRFVRKWR (sequence number 4) (RP837) and FWKKFVKKWK (sequence number 7) (RP841). 9.J 1a , J 1b , J 2a , J 2b , and J. 3a are tryptophan, and X 1a , X 1b , X 2a , X 2b , and X 3a are each independently selected from histidine, lysine, and arginine. 10. Formula:WWX 1a HWWHX 2b WX 3a (SEQ ID NO: 32), wherein X 1a , X 2b , and X 3a is each independently histidine, lysine, or arginine.

[0192] 11. The immunomodulatory peptide of claim 10, having a sequence selected from WWHHWWHHWH (SEQ ID NO: 13), WWRHWWHRWR (SEQ ID NO: 14), and WWKHWWHKWK (SEQ ID NO: 15) (RP847-849). Appendix 12. The striated region is that of Equation 6, and the formula: [J 1a ]-[X 1a X 1b ]-[J 2a J 2b ]-[X 2a ]-[J 3a ]-[X 3a X 3b ]-[J 4a J 4b ] (Formula 6A), wherein J 1a , J 2a , J 2b , J 3a , J 4a , and J. 4b are each independently selected from hydrophobic amino acid residues (e.g., phenylalanine, tryptophan, alanine, isoleucine, valine, and glycine); and X 1a , X 1b , X 2a , X 3a , and X 3b are each independently selected from a hydrophilic amino acid residue (e.g., lysine, arginine, histidine, aspartic acid, glutamic acid, asparagine, or glutamine). 13. The immunomodulatory peptide of claim 12, having the sequence GDRGIKGHRGF (sequence number 8) (RP842). 14. The striated region is that of formula 1, with the formula: [J 1a J 1b ]-[X 1a X 1b ]-[J 2a J 2b ] (Equation 1A) and [J 2b J 2a ]-[X 1b X 1a ]-[J 1b J 1a ] (Formula 1B), wherein J 1a , J 1b , J 2a , and J. 2bare each independently selected from hydrophobic amino acid residues (e.g., phenylalanine, tryptophan, and valine); and X 1a and X 1b are each independently selected from hydrophilic amino acid residues (e.g., lysine or arginine). 15. The immunomodulatory peptide of claim 14, having the sequence FWKRFV (SEQ ID NO: 5) (RP837N).

[0193] 16. The striated region is that of formula 2, and has the formula: [J 1a J 1b ]-[X 1a X 1b ]-[J 2a ]-[X 2a (Formula 2A) wherein J 1a , J 1b , J 2a , and J. 2b are each independently selected from hydrophobic amino acid residues (e.g., phenylalanine, tryptophan, and valine); and X 1a , X 1b , and X 2a are each independently selected from hydrophilic amino acid residues (e.g., lysine or arginine). 17. Array FVRKWR (Array 6) (RP837C 1 17. The immunomodulatory peptide of claim 16, having the following structure: 18. The striated region is that of Equation 4, and has the formula: [J 1a J 1b ]-[X 1a X 1b ]-[J 2a J 2b ]-[X 2a X 2b ]-[J 3a J 3b ] (Equation 4A) and [J 3a J 3b ]-[X 2a X 2b ]-[J 2b J 2a ]-[X 1b X 1a]-[J 1b J 1a ] (Formula 4B), wherein J 1a , J 1b , J 2a , J 2b , J 3a , and J. 3b are each independently selected from a hydrophobic amino acid residue (e.g., phenylalanine, tyrosine, or leucine); and X 1a , X 1b , X 2a , and X 2b are each independently selected from hydrophilic amino acid residues (e.g., lysine or arginine). 19. The immunomodulatory peptide of claim 18, having the sequence LYKKIIKKLL (SEQ ID NO: 12) (RP846). 20. The striated region is that of Equation 4, and the formula: [J 1a J 1b J 1c ]-[X 1a ]-[J 2a J 2b ]-[X 2a X 2b ]-[J 3a J 3b ] (Formula 4C), wherein J 1a , J 1b , J 1c , J 2a , J 2b , J 3a , and J. 3b are each independently selected from a hydrophobic amino acid residue (e.g., phenylalanine, tyrosine, or proline); and X 1a , X 2a , and X 2b are each independently selected from hydrophilic amino acid residues (e.g., aspartic acid, lysine, or arginine).

[0194] 21. The immunomodulatory peptide of claim 20, having the sequence FYPDFFKKFF (SEQ ID NO: 10) (RP844). 22. The striated region is that of Equation 4, and has the formula: [J 1a J 1b ]-[X 1a X 1b ]-[J 2a ]-[X 2a X 2b X 2c ]-[J 3a J 3b (Formula 4D) wherein J 1a , J 1b , J 2a , J 3a , and J. 3b are each independently selected from a hydrophobic amino acid residue (e.g., phenylalanine, serine, glycine, or isoleucine), and X 1a , X 1b , X 2a , X 2b , and X 2c are each independently selected from a hydrophilic amino acid residue (e.g., glutamic acid, aspartic acid, lysine, or arginine). 23. The immunomodulatory peptide of claim 22, having the sequence FFRKSKEKIG (SEQ ID NO: 18) (RP853). 24. The striated region is that of Equation 4, and has the formula: [J 1a J 1b ]-[X 1a X 1b ]-[J 2a J 2b ]-[X 2a X 2b ]-[J 3a ] (Formula 4E), wherein J 1a , J 1b , J 2a , J 2b , and J. 3a are each independently selected from a hydrophobic amino acid residue (e.g., phenylalanine, alanine, or isoleucine); and X 1a , X 1b , X 2a , X 2b , and X 2care each independently selected from hydrophilic amino acid residues (e.g., ornithine, lysine, or arginine). 25. The striated region is that of Equation 5, and the formula: [J 1a ]-[X 1a ]-[J 2a J 2b J 2c ]-[X 2a ]-[J 3a J 3b ]-[X 3a X 3b (Formula 5C) wherein J 1a , J 2a , J 2b , J 2c , J 3a , and J. 3b are each independently selected from a hydrophobic amino acid residue (e.g., phenylalanine, leucine, glycine, or isoleucine), and X 1a , X 2a , X 3a , and X 3b are each independently selected from hydrophilic amino acid residues (e.g., glutamine, lysine, or histidine).

[0195] 26. The immunomodulatory peptide of claim 25, having the sequence FQFLGKIIHH (sequence number 17) (RP852). 27. The striated region is that of Equation 7, and has the formula: [X 1a X 1b ]-[J 1a ]-[X 2a ]-[J 2a ]-[X 3a ]-[J 3a J 3b J 3c ] (Formula 7A), wherein J 1a , J 2a , J 3a , J 3b , and J. 3c are each independently selected from a hydrophobic amino acid residue (e.g., isoleucine, valine, leucine, serine, or alanine); and X1a , X 1b , X 2a , and X 3a are each independently selected from hydrophilic amino acid residues (e.g., lysine or arginine). 28. The immunomodulatory peptide of claim 27, having the sequence KKIRVRLSA (sequence number 16) (RP851). 29. The striated region is that of Equation 5, and the formula: [J 1a J 1b ]-[X 1a X 1b ]-[J 2a J 2b J 2c ]-[X 2b ]-[J 3a ]-[X 3a ] (Formula 5B), wherein J 1a , J 1b , J 2a , J 2b , and J. 3a are each independently selected from a hydrophobic amino acid residue (e.g., phenylalanine, alanine, threonine, or leucine); and X 1a , X 1b , X 2a , X 2b , and X 3a are each independently selected from hydrophilic amino acid residues (e.g., histidine, aspartic acid, lysine, or arginine). 30. The immunomodulatory peptide of claim 29, having the sequence FFRHFATHLD (SEQ ID NO: 11) (RP845).

[0196] 31. The striated region is that of formula 3, and has the formula: [X 1a X 1b ]-[J 1a J 1b J 1c J 1d ]-[X 2a X 2b ]-[J 2a J 2b ] (Formula 3A), wherein J 1a, J 1b , J 1c , J 1d , J 2a , and J. 2b are each independently selected from a hydrophobic amino acid residue (e.g., leucine, serine, alanine, or phenylalanine); 1a , X 1b , X 2a , and X 2b are each independently selected from a hydrophilic amino acid residue (e.g., glutamic acid, aspartic acid, lysine, asparagine, or arginine). 32. The immunomodulatory peptide of claim 31, having the sequence EKLSAFRNFF (sequence number 9) (RP843). 33. The immunomodulatory peptide of Appendix 1, wherein the stripathic region comprises a dimer of a first and a second polypeptide connected via a peptide linker connecting the C-terminus of the first polypeptide and the N-terminus of the second polypeptide. 34. The immunomodulatory peptide of claim 33, wherein the hydrophilic module consists of amino acid residues selected from lysine, arginine, and ornithine, and the hydrophobic module consists of amino acid residues selected from phenylalanine and tryptophan. 35. The immunomodulatory peptide of claim 33 or 34, wherein the first and second polypeptides comprise one of the following formulas: [X1]-[J1]-[X2]-[J2] (Formula 3) or [J1]-[X1]-[J2]-[X2] (Formula 2).

[0197] 36. The immunomodulatory peptide of any one of appendices 33 to 35, wherein the dimer has one of the following formulas: [X1]-[J1]-[X2]-[J2]-T-[J2]-[X2]-[J1]-[X1] or [J1]-[X1]-[J2]-[X2]-T-[X2]-[J2]-[X1]-[J1], wherein T is a peptide linker. 37. The dimer has the following formula: [X 1a ]-[J 1a ]-[X 2a ]-[J 2a ]-T-[J 2a]-[X 2a ]-[J 1a ]-[X 1a ] (Equation 12A) or [J 1a ]-[X 1a ]-[J 2a ]-[X 2a ]-T-[X 2a ]-[J 2a ]-[X 1a ]-[J 1a ] (Formula 13A), wherein T is a peptide linker (e.g., a polyglycine linker). 38. The immunomodulatory peptide according to appended claim 37, having a sequence selected from RWKFGGFKWR (SEQ ID NO: 1) (RP832C) and FKWRGGRWKF (SEQ ID NO: 3) (RP837C). 39. The dimer has the following formula: [X 1a X 1b ]-[J 1a J 1b ]-T-[J 1b J 1a ]-[X 1b X 1a ] (Equation 8A) or [J 1a J 1b ]-[X 1a X 1b ]-T-[X 1b X 1a ]-[J 1b J 1a ] (Formula 9A), wherein T is a peptide linker (e.g., a polyglycine linker), and J 1a and J. 1b are each independently selected from hydrophobic amino acid residues (e.g., tryptophan or phenylalanine); 1a and X 1b 35. The immunomodulatory peptide of claim 33 or 34, wherein each is independently selected from a hydrophilic amino acid residue (e.g., asparagine or arginine). 40. The immunomodulatory peptide of appendix 39 having the sequence FWKRGGRKWF (sequence number 4) (peptide 837A).

[0198] 41. a) a sequence selected from the peptide sequences of Table 3; b) a sequence having at least 75% sequence identity (e.g., at least 80%, at least 85%, at least 90%, or at least 95% sequence identity) with a sequence defined in a); or c) a sequence having one or two amino acid substitutions compared to the sequence defined in a), wherein the one or two amino acid substitutions are amino acid substitutions according to Table 2 (e.g., similar, conservative, or highly conservative amino acid substitutions). 41. The immunomodulatory peptide according to any one of appendices 1 to 40, comprising: 42. An immunomodulatory peptide according to any one of appendices 1 to 41, consisting of a sequence selected from any one of the sequences in Table 3 (SEQ ID NOs: 1 to 19).

[0199] 43. An immunomodulatory peptide having a length of 6 to 30 amino acid residues, a) a peptide sequence selected from SEQ ID NOs: (1-19) (e.g., RP832C, 837, 837A, 837C, 837N, 841-842, 843-850, and 853), or b) a sequence having one or two amino acid substitutions compared to the sequence defined in a), wherein the one or two amino acid substitutions are amino acid substitutions according to Table 2 (e.g., similar, conservative, or highly conservative amino acid substitutions). 1. An immunomodulatory peptide comprising: 44. The immunomodulatory peptide of claim 43, wherein one or two amino acid substitutions as defined in clause b) consist of a substitution of a cationic amino acid of the sequence with an alternative cationic amino acid residue (e.g., O for K, K for O, R for K, etc.). 45. The immunomodulatory peptide of claim 43, comprising a peptide sequence selected from: RWKFGGFKWR (RP832C) (SEQ ID NO: 1), FKWRGGRWKF (RP837C) (SEQ ID NO: 3), and FWKRGGRKWF (RP837A) (SEQ ID NO: 4).

[0200] 46. ​​FWKRFV (RP837N) (SEQ ID NO: 5) and FVRKWR (RP837C 1 44. The immunomodulatory peptide of claim 43, comprising a peptide sequence selected from: 47. The immunomodulatory peptide of claim 43, comprising a peptide sequence selected from FAOOFAOOFO (RP850) (SEQ ID NO: 19), FWKRFVRKWR (RP837) (SEQ ID NO: 4), and FWKKFVKKWK and (RP841) (SEQ ID NO: 7). 48. The immunomodulatory peptide of claim 43, comprising a peptide sequence selected from WWHHWWHHWH (SEQ ID NO: 13), WWRHWWHRWR (SEQ ID NO: 14), and WWKHWWHKWK (SEQ ID NO: 15) (RP847-849). 49. The immunomodulatory peptide of claim 43, comprising the peptide sequence GDRGIKGHRGF(RP842) (sequence number 8). 50. The immunomodulatory peptide of claim 43, comprising the peptide sequence LYKKIIKKLL(RP846) (SEQ ID NO: 12).

[0201] 51. The immunomodulatory peptide of claim 43, comprising the peptide sequence FYPDFFKKFF(RP844) (SEQ ID NO: 10). 52. The immunomodulatory peptide of claim 43, comprising the peptide sequence FFRKSKEKIG (RP853) (SEQ ID NO: 18). 53. The immunomodulatory peptide of claim 43, comprising the peptide sequence FFRHFATHLD(RP845) (SEQ ID NO: 11). 54. The immunomodulatory peptide of claim 43, comprising the peptide sequence EKLSAFRNFF(RP843) (SEQ ID NO: 9).

[0202] 55. An immunomodulatory peptide (e.g., 12 amino acid residues or less in length), An immunomodulatory peptide comprising a sequence selected from RWKFGGFKWR (RP832C) (SEQ ID NO: 1), FKWRGGRWKF (RP837C) (SEQ ID NO: 3), and FWKRGGRKWF (RP837A) (SEQ ID NO: 4). 56. The immunomodulatory peptide according to appendix 55, consisting of the sequence RWKFGGFKWR (RP832C) (SEQ ID NO: 1). 57. The immunomodulatory peptide according to appendix 55, consisting of the sequence FKWRGGRWKF(RP837C) (sequence number 3). 58. The immunomodulatory peptide according to appendix 55, consisting of the sequence FWKRGGRKWF(RP837A) (sequence number 4).

[0203] 59. A pharmaceutical composition comprising an immunomodulatory peptide according to any one of appendices 1 to 58 and a pharmaceutically acceptable carrier. 60. The pharmaceutical composition of claim 59, wherein the composition is formulated for oral administration, parenteral administration, administration via inhalation, or topical administration. 61. The pharmaceutical composition of claim 59 or 60, wherein the composition is formulated for intravenous or subcutaneous administration. 62. The pharmaceutical composition of claim 59 or 60, wherein the composition is formulated for oral administration and further comprises an enteric coating. 63. The pharmaceutical composition of claim 59 or 60, wherein the composition is formulated for topical delivery in a form selected from the group consisting of a gel suspension, a cream, a microneedle, and is injected into a bandage or a topical patch.

[0204] 64. A method for modulating macrophage activity, comprising: A method comprising contacting a macrophage with a CD206-binding agent to modulate the activity of the macrophage. 65. The method of claim 64, wherein the CD206-binding agent binds to the mannose-binding site and modulates signal-regulatory protein (SIRP)-mannose binding to CD206. 66. The method of claim 64 or 65, wherein the CD206-binding agent binds to CD206 with a binding energy of at least -650 kcal / mol. 67. The method of any one of clauses 64 to 66, wherein the CD206-binding agent directly contacts at least one amino acid residue of CD206 selected from Phe-708, Thr-709, Trp-710, Pro-714, Glu-719, Asn-720, Trp-721, Ala-722, Glu-725, Tyr-729, Glu-733, Asn-747, Asp-748, Ser-1691, Cys-1693, Phe-1694, and Phe-1703. 68. The method of any one of claims 64 to 67, wherein the modulated macrophage activity is macrophage polarization. 69. A method according to any one of claims 64 to 68, wherein the survival rate of macrophages is reduced. 70. The method of any one of claims 64 to 69, wherein the macrophages are M2 macrophages or tumor-associated macrophages (TAMs).

[0205] 71. The method of any one of clauses 64 to 70, wherein the CD206-binding agent inhibits macrophage activity. 72. The method of any one of clauses 64 to 71, wherein the CD206-binding agent is an immunomodulatory peptide. 73. The method of any one of claims 64 to 71, wherein the macrophages are in vitro. 74. The method of any one of claims 64 to 71, wherein the macrophages are in vivo. 75. The method of any one of appendices 64 to 74, wherein the CD206-binding agent is an immunomodulatory peptide according to any one of appendices 1 to 58.

[0206] 76. A method of treating a subject for a condition associated with chronic inflammation, comprising: A method comprising administering to a subject an effective amount of a CD206-binding agent (e.g., an immunomodulatory peptide described in any one of Appendices 1-58) to treat the subject for a condition associated with chronic inflammation. 77. The method of claim 76, wherein the condition associated with chronic inflammation is selected from the group consisting of scleroderma or multiple sclerosis, irritable bowel disease, ulcerative colitis, colitis, Crohn's disease, idiopathic pulmonary fibrosis, scleroderma, asthma, keratitis, arthritis, osteoarthritis, rheumatoid arthritis, autoimmune diseases, feline or human immunodeficiency virus (FIV or HIV) infection, cancer, age-related inflammation and / or stem cell dysfunction, graft-versus-host disease (GVHD), keloids, obesity, diabetes, diabetic wounds, other chronic wounds, atherosclerosis, Parkinson's disease, Alzheimer's disease, macular degeneration, gout, gastric ulcer, gastritis, mucositis, toxoplasmosis, and chronic viral or microbial infections. 78. The method of claim 76 or 77, wherein the CD206-binding agent is an immunomodulatory peptide according to any one of claims 1 to 58. 79. The method of claim 77, wherein the pathological condition is cancer. 80. The method of claim 79, further comprising administering to the subject an effective amount of an additional agent.

[0207] 81. The method of claim 80, wherein the additional agent is a chemotherapeutic agent. 82. The method of claim 80, wherein the chemotherapeutic agent is selected from gemcitabine, docetaxel, bleomycin, erlotinib, gefitinib, lapatinib, imatinib, dasatinib, nilotinib, bosutinib, crizotinib, ceritinib, trametinib, bevacizumab, sunitinib, sorafenib, trastuzumab, ado-trastuzumab emtansine, rituximab, ipilimumab, rapamycin, temsirolimus, everolimus, methotrexate, doxorubicin, Abraxane, Forfirinox, cisplatin, carboplatin, 5-fluorouracil, Teismo, paclitaxel, prednisone, levothyroxine, and pemetrexed. 83. The method of claim 81, wherein the chemotherapeutic agent is Abraxane. 84. The method of claim 81, wherein the chemotherapeutic agent is gemcitabine or docetaxel. 85. The method of claim 80, wherein the additional agent is an immunotherapeutic agent.

[0208] 86. The method of claim 85, wherein the immunotherapeutic agent is an immune checkpoint inhibitor. 87. The method of claim 86, wherein the immune checkpoint inhibitor is selected from a cytotoxic T-lymphocyte-associated antigen 4 (CTLA-4) inhibitor, a programmed death 1 (PD-1) inhibitor, and a PD-L1 inhibitor. 88. The method of claim 87, wherein the immune checkpoint inhibitor is selected from ipilimumab, pembrolizumab, and nivolumab. 89. The method according to claim 76, wherein the pathology associated with chronic inflammation is fibrosis. 90. The method according to claim 76, wherein the pathology associated with chronic inflammation is scleroderma.

[0209] 91. The method of any one of appendices 76 to 90, wherein the CD206-binding agent is an immunomodulatory peptide according to any one of appendices 1 to 58. 92. The method of claim 91, wherein the CD206-binding agent consists of an immunomodulatory peptide of Table 3.

Claims

1. An immunomodulatory peptide having 12 or less amino acids, a) the peptide sequence RWKFGGFKWR (RP832C) (SEQ ID NO: 1); b) the peptide sequence FWKRFVRKWR (RP837) (SEQ ID NO: 2), or c) a sequence which has one or two amino acid substitutions compared to the sequences defined in a) or b), wherein said one or two amino acid substitutions consist of highly conservative substitutions of cationic amino acids of said sequences.

1. An immunomodulatory peptide comprising:

2. 2. The immunomodulatory peptide of claim 1, comprising the peptide sequence FWKRFVRKWR (RP837) (SEQ ID NO: 2).

3. 2. The immunomodulatory peptide of claim 1, comprising the peptide sequence RWKFGGFKWR (RP832C) (SEQ ID NO: 1).

4. A pharmaceutical composition comprising the immunomodulatory peptide of claim 1 and a pharmaceutically acceptable carrier.

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