EP4652181A1

EP4652181A1 - Polycyclic peptide inhibitors of interleukin-23 receptor

Info

Publication number: EP4652181A1
Application number: EP24706606.1A
Authority: EP
Inventors: Santhosh Francis Neelamkavil; Chengzao Sun; Sandeep Somani; Jing Zhang; Danila Branca; Roberto COSTANTE; Elisabetta Bianchi; Raffaele Ingenito; Stephanie A. Barros
Original assignee: Janssen Pharmaceutica NV
Current assignee: Janssen Pharmaceutica NV
Priority date: 2023-01-16
Filing date: 2024-01-15
Publication date: 2025-11-26
Also published as: JP2026503109A; WO2024155551A1; CN121127485A

Abstract

The present disclosure relates to peptide inhibitors of the interleukin-23 receptor (IL- 23R) or pharmaceutically acceptable salts thereof, corresponding pharmaceutical compositions, methods and/or uses for treatment of autoimmune inflammation and related diseases and disorders.

Description

POLYCYCLIC PEPTIDE INHIBITORS OF INTERLEUKIN-23 RECEPTOR

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional Application No. 63/480,042 filed January 16, 2023, which is herein incorporated by reference in its entirety.

INCORPORATION OF SEQUENCE LISTING

The sequence listing in ST.26 XML format entitled 739655 NTT-4252PC SL, created on January 13, 2024, comprising 29,940 bytes, prepared according to 37 CFR 1.822 to 1.824, submitted concurrently with the filing of this application, is incorporated herein by reference in its entirety.

BACKGROUND

The interleukin-23 (IL-23) cytokine is a heterodimer composed of a unique pl9 subunit and the p40 subunit shared with IL-12, which is a cytokine involved in the development of interferon-y (IFN-y)-producing T helper 1 (THI) cells. Although IL-23 and IL-12 both contain the p40 subunit, they have different phenotypic properties. For example, animals deficient in IL- 12 are susceptible to inflammatory autoimmune diseases, whereas IL-23 deficient animals are resistant to these diseases, presumably due to a reduced number of CD4⁺ T cells producing IL-6, IL-17, and TNF in the CNS of IL-23 -deficient animals. IL-23 binds to IL-23R, which is a heterodimeric receptor composed of IL-12RP1 and IL-23R subunits. Binding of IL-23 to IL-23R activates the Jak-Stat signaling molecules Jak2, Tyk2, Statl, Stat 3, Stat 4, and Stat 5, although Stat4 activation is substantially weaker and different DNA-binding Stat complexes form in response to IL-23 as compared with IL-12. IL-23R associates constitutively with Jak2 and in a ligand-dependent manner with Stat3. In contrast to IL-12, which acts mainly on naive CD4(+) T cells, IL-23 preferentially acts on memory CD4(+) T cells.

IL-23 has been implicated as playing a crucial role in the pathogenesis of autoimmune inflammation and related diseases and disorders, such as multiple sclerosis, asthma, rheumatoid arthritis, psoriasis, and inflammatory bowel diseases (IBDs) such as ulcerative colitis and Crohn’s disease. Studies in acute and chronic mouse models of IBDs revealed a primary role of interleukin-23 receptor (IL-23R) and downstream effector cytokines in disease pathogenesis. IL-23R is expressed on various adaptive and innate immune cells including Thl7 cells, y5 T cells, natural killer (NK) cells, dendritic cells, macrophages, and innate lymphoid cells, which are found abundantly in the intestine. At the intestine mucosal surface, the gene expression and protein levels of IL-23R are found to be elevated in IBD patients. It is believed that IL-23 mediates this effect by promoting the development of a pathogenic CD4⁺ T cell population that produces IL-6, IL-17, and tumor necrosis factor (TNF). Accordingly, there remains a need for compositions that bind IL-23R to inhibit IL-23 binding and signaling in a patient.

BRIEF SUMMARY

Provided herein are peptide inhibitors of the interleukin-23 receptor (IL-23R) or pharmaceutically acceptable salts thereof, corresponding pharmaceutical compositions, and methods and/or uses of the IL-23R inhibitors for the treatment of inflammatory diseases, autoimmune diseases, and/or related disorders.

In particular, the present disclosure provides a peptide of Formula (I’), comprising the amino acid sequence:

Ri-X3-X4-X5-X6-X7-X₈-X₉-Xio-2Nal-Xi2-Xi3-N-Xi5-Xi6-R2 (F), or a pharmaceutically acceptable salt thereof, wherein each of X3, X4, X5, Xe, X7, X₈, X9, X10, X12, X13, X15, and Xi6 are defined herein.

In some embodiments, the present disclosure provides a peptide of Formula (I), comprising the amino acid sequence:

Ri-X3-X4-X₅-T-X7-X₈-X₉-Xio-2Nal-Xi2-Xi3-N-Xi₅-Xi6-R2 (I), or a pharmaceutically acceptable salt thereof, wherein each of X3, X₄, X5, X7, X₈, X9, X10, X12, X13, X15, and Xi6 are defined herein.

The present disclosure further provides a pharmaceutical composition comprising a peptide described herein, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

The present disclosure still further provides a method for treating a disease or disorder associated with Interleukin 23 (IL-23)/Interleukin 23 Receptor (IL-23R), comprising administering to a subject in need thereof a therapeutically effective amount of a peptide or a pharmaceutical composition described herein. In some embodiments, the disease or disorder is selected from ulcerative colitis (UC), Crohn’s disease (CD), psoriasis (PsO), and psoriatic arthritis (PsA).

DETAILED DESCRIPTION

Provided herein are peptide inhibitors of IL-23R, and pharmaceutically acceptable salts thereof, corresponding pharmaceutical compositions, and methods and/or uses for the treatment of inflammatory diseases, autoimmune diseases, and/or related disorders. Definitions

Unless otherwise defined herein, scientific and technical terms used in this application shall have the meanings that are commonly understood by those of ordinary skill in the art.

As used in the specification and in the claims, the “comprise(s),” “comprising,” “include(s),” “having,” “has,” “can,” “contain(s),” and variants thereof, as used herein, are intended to be open-ended transitional phrases, terms, or words that require the presence of the named features, groups, ingredients, or steps and does not exclude the presence of additional features, groups, ingredients, or steps. For example, the language “a peptide of Formula (I), comprising the amino acid sequence: X3-X4-X5-T-X7-X8-X9-Xio-2Nal-Xi2-Xi3-N-Xi5-Xi6 (I),” means that in addition to amino acids X3 through X17, the peptide may include but is not limited to additional amino acids attached to the N-terminus, additional amino acids attached to the C- terminus, N-terminal or C-terminal capping groups, chemical or biological moieties (including but not limited to, for example, lipophilic substituents, antibodies, imaging agents, etc.) conjugated to the peptide at any location, and the like. The term “comprise(s),” “comprising,” “include(s),” “having,” “has,” “can,” or “contain(s),” can include embodiments encompassed by the term "consisting essentially of or "consisting of."

The terms “peptide,” “polypeptide,” and “protein” are used interchangeably herein and typically refer to a molecule comprising a chain of two or more amino acids (e.g., L-amino acids, D-amino acids, modified amino acids, amino acid analogs, amino acid mimetics, etc.).

Unless otherwise indicated, naturally-occurring L-amino acids and D-amino acids are both represented by either conventional three-letter, or capitalized one-letter, amino acid designations of Table 1. In some embodiments, naturally-occurring L-amino acids are represented by either conventional three-letter, or capitalized one-letter, amino acid designations of Table 1. In some embodiments, D-amino acids, are represented by lower-case one-letter amino acid designations corresponding to one-letter designations of Table 1, i.e., g, a, 1, m, f, w, k, q, e, s, p, v, i, c, y, h, r, n, d, and t.

Table 1: Naturally-occurring amino acids

G Glycine Gly P Proline Pro A Alanine Ala V Valine Vai L Leucine Leu I Isoleucine He M Methionine Met C Cysteine Cys F Phenylalanine Phe Y Tyrosine Tyr W Tryptophan Trp H Histidine His K Lysine Lys R Arginine Arg

Q Glutamine Gin N Asparagine Asn E Glutamic Acid Glu D Aspartic Acid Asp S Serine Ser T Threonine Thr

The term “L-amino acid,” as used herein, refers to the “L” isomeric form of an amino acid, and conversely the term “D-amino acid” refers to the “D” isomeric form of an amino acid (e.g., (D)Asp or D-Asp; (D)Phe or D-Phe). Amino acid residues in the D isomeric form can be substituted for any L-amino acid residue, as long as the desired function is retained by the peptide. D-amino acids may be indicated as customary in lower case when referred to using single-letter abbreviations. For example, D-arginine can be represented as “arg” or “r.” Alternatively, a lower case “d” in front of an amino acid can be used to indicate that it is of the D isomeric form, for example D-lysine can be represented by dK.

In the case of less common or non-naturally occurring amino acids, unless they are referred to by their full name (e.g., sarcosine, ornithine, etc.), frequently employed three- or four-character codes are employed for residues thereof, including, Sar or Sarc (sarcosine, i.e., N- methylglycine), Aib (a-aminoisobutyric acid), Dab (2,4-diaminobutanoic acid), Dapa (2,3- diaminopropanoic acid), y-Glu /-glutamic acid), Gaba (y-aminobutanoic acid), P-Pro (pyrrolidine-3 -carboxylic acid), and Abu (2-aminobutyric acid).

Amino acids of the D-isomeric form may be located at any of the positions in the IL- 23R inhibitors set forth herein (e.g., any of X3-X16 appearing in the molecule). In some embodiments, amino acids of the D-isomeric form may be located only at any one or more of X3, X5, Xe, Xs, X13, and optionally one additional position. In other embodiments, amino acids of the D-isomeric form may be located only at any one or more of X3, Xs, X13, and optionally one additional position. In other embodiments, amino acids of the D-isomeric form may be located only at any one or more of Xs, X13, and optionally one additional position. In other embodiments, amino acids of the D-isomeric form may be located only at X3 and optionally one additional position. In other embodiments, amino acids of the D-isomeric form may be located only at X3, and optionally two or three additional positions. In other embodiments, amino acids of the D-isomeric form may be located at only one or two of positions X3 to Xi6 appearing in the IL-23R inhibitors set forth herein. In other embodiments, amino acids of the D-isomeric form may be located at only three or four of positions X3 to Xi6 appearing in the IL-23R inhibitors set forth herein. For example, an IL-23R inhibitor set forth herein having only positions X3 to X15 present may have amino acids of the D-form present in three or four of those positions. In other embodiments, amino acids of the D-isomeric form may be located at only five or six of positions X3 to X17 appearing in the IL-23R inhibitors set forth herein. In other embodiments, amino acids of the D-isomeric form may be located at no more than one of positions X3 to Xi6 appearing in the IL-23R inhibitors set forth herein. Peptides may be naturally occurring, synthetically produced, or recombinantly expressed. Peptides may also comprise additional groups modifying the amino acid chain, for example, functional groups added via post-translational modification. Examples of post-translation modifications include, but are not limited to, acetylation, alkylation (including, methylation), biotinylation, glutamylation, glycylation, glycosylation, isoprenylation, lipoylation, phosphopantetheinylation, phosphorylation, selenation, and C-terminal amidation. The term peptide also includes peptides comprising modifications of the amino terminus and/or the carboxy terminus. Modifications of the terminal amino group include, but are not limited to, desamino, N-lower alkyl, N-di-lower alkyl, and N-acyl modifications. Modifications of the terminal carboxy group include, but are not limited to, amide, lower alkyl amide, dialkyl amide, and lower alkyl ester modifications (e.g., wherein lower alkyl is C1-C4 alkyl). The term peptide also includes modifications, such as but not limited to those described above, of amino acids falling between the amino and carboxy termini.

As is clear to the skilled artisan, the peptide sequences disclosed herein are shown proceeding from left to right, with the left end of the sequence being the N-terminus of the peptide and the right end of the sequence being the C-terminus of the peptide. Among sequences disclosed herein are sequences incorporating either an “-OH” moiety or an “-NH2” moiety at the carboxy terminus (C-terminus) of the sequence. In such cases, and unless otherwise indicated, an “-OH” or an “-NH2” moiety at the C-terminus of the sequence indicates a hydroxy group or an amino group, corresponding to the presence of a carboxylic acid (COOH) or an amido (CONH2) group at the C-terminus, respectively. In each sequence of the disclosure, a C-terminal “-OH” moiety may be substituted for a C-terminal “-NH2” moiety, and vice-versa.

The phrase “amino acid,” “amino acid residue,” or “residue” as used herein refers to an amino acid, a modified amino acid, an amino acid analog, or an amino acid mimetic that is incorporated into a peptide by an amide bond or an amide bond mimetic.

Unless indicated otherwise the names of naturally occurring and non-naturally occurring aminoacyl residues used herein follow the naming conventions suggested by the IUPAC Commission on the Nomenclature of Organic Chemistry and the IUPAC-IUB Commission on Biochemical Nomenclature as set out in “Nomenclature of a-Amino Acids (Recommendations, 1974)” Biochemistry, 14(2), (1975). To the extent that the names and abbreviations of amino acids and aminoacyl residues employed in this specification and appended claims differ from those suggestions, they will be made clear to the reader. In sequences of amino acids that represent IL-23 inhibitors the individual amino acids are separated by a hyphen or brackets e.g, lysine is shown as [K], One of skill in the art will appreciate that certain amino acids and other chemical moi eties are modified when bound to another molecule. For example, an amino acid side chain may be modified when it forms an intramolecular bridge with another amino acid side chain, e.g., one or more hydrogens may be removed or replaced by the bond.

The term “therapeutically effective amount” or “pharmaceutically effective amount” means that amount of active peptide or pharmaceutical agent that elicits the biological or medicinal response in a tissue system, animal or human, that is being sought by a researcher, veterinarian, medical doctor, or other clinician, which includes preventing, treating or ameliorating the symptoms of a syndrome, disorder or disease being treated.

The term “pharmaceutically acceptable” means approved or approvable by a regulatory agency of Federal or a state government or the corresponding agency in countries other than the United States, or that is listed in the U. S. Pharmcopoeia or other generally recognized pharmacopoeia for use in animals, and more particularly, in humans.

“Pharmaceutically acceptable excipient” includes without limitation any adjuvant, carrier, excipient, glidant, sweetening agent, diluent, preservative, dye/colorant, flavor enhancer, surfactant, wetting agent, dispersing agent, suspending agent, stabilizer, isotonic agent, solvent, or emulsifier which has been approved by the United States Food and Drug Administration as being acceptable for use in humans or domestic animals.

“Composition” or “pharmaceutical composition” as used herein is intended to encompass a product comprising the specified active pharmaceutical ingredient (API) (i.e., a peptide of the present disclosure), which may include pharmaceutically acceptable excipients, carriers or diluents as described herein, such as in specified amounts defined throughout the disclosure.

Compositions or pharmaceutical compositions of the present disclosure may be in different pharmaceutically acceptable forms, which may include, but are not limited to a liquid composition, a tablet or matrix composition, a capsule composition, etc.. When the composition is a tablet composition, the tablet may include, but is not limited to different layers two or more different phases, including an internal phase and an external phase that can comprise a core. The tablet composition can also include, but is not limited to one or more coatings.

Provided are also pharmaceutically acceptable salts and tautomeric forms of the peptides described herein.

A “pharmaceutically acceptable salt” is intended to mean a salt of a free acid or base of peptides represented by Formula (I) that are non-toxic, biologically tolerable, or otherwise biologically suitable for administration to the subject. It should possess the desired pharmacological activity of the parent compound. See, generally, G.S. Paulekuhn, et al., “Trends in Active Pharmaceutical Ingredient Salt Selection based on Analysis of the Orange Book Database”, J. Med. Chem., 2007, 50:6665-72, S.M. Berge, et al., “Pharmaceutical Salts”, J Phctrm Sci .. 1977, 66: 1-19, and Handbook of Pharmaceutical Salts, Properties, Selection, and Use, Stahl and Wermuth, Eds., Wiley-VCH and VHCA, Zurich, 2002. Examples of pharmaceutically acceptable salts are those that are pharmacologically effective and suitable for contact with the tissues of patients without undue toxicity, irritation, or allergic response. A peptide of Formula (I) may possess a sufficiently acidic group, a sufficiently basic group, or both types of functional groups, and accordingly react with a number of inorganic or organic bases, and inorganic and organic acids, to form a pharmaceutically acceptable salt.

The IL-23R inhibitors of the present disclosure, pharmaceutically acceptable salts, and/or other forms thereof may contain one or more asymmetric centers and may thus give rise to enantiomers, diastereomers, and other stereoisomeric forms that may be defined, in terms of absolute stereochemistry, as (R)- or (5)- or, as (D)- or (L)- for amino acids. The present disclosure is meant to include all such possible isomers, as well as their racemic and optically pure forms of the IL-23R inhibitors of the present disclosure. Optically active (+) and (-), (R)- and (5)-, or (D)- and (L)- isomers may be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques, for example, chromatography and fractional crystallization. Conventional techniques for the preparation/isolation of individual enantiomers include chiral synthesis from a suitable optically pure precursor or resolution of the racemate (or the racemate of a salt or derivative) using, for example, chiral high pressure liquid chromatography (HPLC). When the compounds described herein contain olefinic double bonds or other centers of geometric asymmetry, and unless specified otherwise, it is intended that the compounds include both E and Z geometric isomers. Likewise, all tautomeric forms are also intended to be included. Where compounds are represented in their chiral form, it is understood that the aspect encompasses, but is not limited to, the specific diastereomerically or enantiomerically enriched form. Where chirality is not specified but is present, it is understood that the aspect is directed to either the specific diastereomerically or enantiomerically enriched form; or a racemic or scalemic mixture of such compound(s).

“Racemates” refers to a mixture of enantiomers. The mixture can include equal or unequal amounts of each enantiomer.

“Stereoisomer” and “stereoisomers” refer to compounds that differ in the chirality of one or more stereo centers. Stereoisomers include enantiomers and diastereomers. The compounds may exist in stereoisomeric form if they possess one or more asymmetric centers or a double bond with asymmetric substitution and, therefore, can be produced as individual stereoisomers or as mixtures. Unless otherwise indicated, the description is intended to include individual stereoisomers as well as mixtures. The methods for the determination of stereochemistry and the separation of stereoisomers are well-known in the art (see, e.g., Chapter 4 of Advanced Organic Chemistry, 4th ed., J. March, John Wiley and Sons, New York, 1992).

“Diastereoisomers” are stereoisomers that have at least two asymmetric atoms, but which are not mirror images of each other.

“Enantiomers” are a pair of stereoisomers that are non-superimposable mirror images of each other. A “racemic” mixture is a 1 : 1 mixture of a pair of enantiomers. A “scalemic” mixture of enantiomers is mixture of enantiomers at a ratio other than 1 : 1.

“Tautomer” refers to alternate forms of a compound that differ in the position of a proton, such as enol -keto and imine-enamine tautomers, or the tautomeric forms of heteroaryl groups containing a ring atom attached to both a ring -NH- and a ring =N- such as pyrazoles, imidazoles, benzimidazoles, triazoles, and tetrazoles.

The term “administering” with respect to the methods of the present disclosure, means a method for therapeutically or prophylactically preventing, treating or ameliorating a syndrome, disorder or disease as described herein by using a compound of the disclosure, or pharmaceutically acceptable salt thereof, composition thereof, or medicament thereof. Such methods include administering a therapeutically effective amount of a peptide of the disclosure, or pharmaceutically acceptable salt thereof, composition thereof, or medicament thereof, at different times during the course of a therapy or concurrently or sequentially as a combination therapy.

“Patient” or “subject”, which are used interchangably, refer to a living organism, preferably a mammal, most preferably a human, whom will be or has been treated by a method according to an embodiment of the application. Examples of mammals include, but are not limited to, cows, horses, sheep, pigs, cats, dogs, mice, rats, rabbits, guinea pigs, non-human primates (NHPs) such as monkeys or apes, humans, etc., more preferably a human.

As used herein, the term “treatment” or “treating,” is defined as the application or administration of a therapeutic agent, i.e., a compound of the present disclosure (alone or in combination with another pharmaceutical agent), to a patient, or application or administration of a therapeutic agent to an isolated tissue or cell line from a patient (e.g., for diagnosis or ex vivo applications), who has a disorder or disease as described herein, a symptom thereof; or the potential to develop such disorder or disease, where the purpose of the application or administration is to cure, heal, alleviate, relieve, alter, remedy, ameliorate, improve or affect the disorder or disease, its symptoms, or the potential to develop said disorder or disease. Such treatments may be specifically tailored or modified, based on knowledge obtained from the field of pharmacogenomics. As used herein, the term “prevent” or “prevention” means no disorder or disease development if none had occurred, or no further disorder or disease development if there had already been development of the disorder or disease. Also considered is the ability of one to prevent some or all of the symptoms associated with the disorder or disease.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly or conventionally understood by one of ordinary skill in the art. In the chemical arts a dash at the front or end of a chemical group is a matter of convenience; chemical groups may be depicted with or without one or more dashes without losing their ordinary meaning. A wavy line drawn through a line in a structure indicates a point of attachment of a group. A dashed line indicates an optional bond. Unless chemically or structurally required, no directionality is indicated or implied by the order in which a chemical group is written or the point at which it is attached to the remainder of the molecule. For instance, the group “-SO2CH2- ” is equivalent to “-CH2SO2-” and both may be connected in either direction. Similarly, an “arylalkyl” group, for example, may be attached to the remainder of the molecule at either an aryl or an alkyl portion of the group. A prefix such as “C_u-v” or (C_u-C_v) indicates that the following group has from u to v carbon atoms. For example, “Ci-ealkyl” and “Ci-Ce alkyl” both indicate that the alkyl group has from 1 to 6 carbon atoms.

Furthermore, it is intended that within the scope of the present invention, any element, in particular when mentioned in relation to a peptide of the disclosure, or pharmaceutically acceptable salt thereof, shall comprise all isotopes and isotopic mixtures of said element, either naturally occurring or synthetically produced, either with natural abundance or in an isotopically enriched form. For example, a reference to hydrogen includes within its scope ¹H, ²H (z.e., deuterium or D), and ³H (z.e., tritium or T). In some embodiments, the compounds described herein include a ²H (z.e., deuterium) isotope. By way of example, the group denoted -C(i-6)alkyl includes not only -CH3, but also CD3; not only CH2CH3, but also CD2CD3, etc. Similarly, references to carbon and oxygen include within their scope respectively ¹²C, ¹³C and ¹⁴C and ¹⁵O and ¹⁶O and ¹⁷O and ¹⁸O. The isotopes may be radioactive or non-radioactive. Radiolabelled compounds of the disclsoure may include a radioactive isotope selected from the group comprising ³H, ⁿC, ¹⁸F, ³⁵S, ¹²²I, ¹²³I, ¹²⁵I, ¹³¹I, ⁷⁵Br, ⁷⁶Br, ⁷⁷Br and ⁸²Br. Preferably, the radioactive isotope is selected from the group of ³H, ⁿC and ¹⁸F.

Abbreviation, “(V/V)” refers to the phrase “volume for volume”, i.e., the proportion of a particular substance within a mixture, as measured by volume or a volume amount of a component of the composition disclosed herein relative to the total volume amount of the composition. Accordingly, the quantity is unit less and represents a volume percentage amount of a component relative to the total volume of the composition. For example, a 2% (V/V) solvent mixture can indicate 2 mL of one solvent is present in 100 mL of the solvent mixture.

“Bioavailability” refers to the extent and rate at which the active moiety (drug or metabolite) enters systemic circulation, thereby accessing the site of action. Bioavailability of a drug could be impacted by the factors such as properties of the dosage form and properties of the drug

“Digestive tract tissue” as used herein refers to all the tissues that comprise the organs of the alimentary canal. For example only, and without limitation, “digestive tract tissue” includes tissues of the mouth, esophagus, stomach, small intestine, large intestine, duodenum, and anus.

Compounds

The present disclosure provides a peptide inhibitor of interleukin-23 receptor. In particular, the present disclosure provides a peptide of Formula (F), comprising the amino acid sequence:

Ri-X3-X4-X5-X6-X7-X₈-X₉-Xio-2Nal-Xi2-Xi3-N-Xi5-Xi6-R2 (F), or a pharmaceutically acceptable salt thereof, wherein:

Ri is MeCO, 8Aoc, 7Ahp, 6Ahx, 5Ava, or cPEG3aCO;

X3 is hK, a ring-forming amino acid, or absent;

X4 is any amino acid;

X5 is N, N(NMe2), Q, Q(NMe2), or a ring-forming amino acid;

Xe is any amino acid;

X₇ is 7MeW or W;

X₈ is K(Ac), K(NMeAc), Q, or a ring-forming amino acid;

X9 is any amino acid;

X10 is AEF, APEG3F, F(4TzlAme2), TMAPF, or a ring-forming amino acid;

X12 is THP, or a ring-forming amino acid;

X13 is E or a ring-forming amino acid;

X15 is 3Pya, bAla, or a ring-forming amino acid;

Xi6 is Sar, a ring-forming amino acid, or absent;

R₂ is CONH2 or CONMe₂; wherein:

(a) a first ring-forming amino acid is linked to a second-ring forming amino acid to form a first ring comprising 4-11 or 14 amino acids; and a third ring-forming amino acid is linked to a fourth ring forming amino acid to form a second ring comprising 4-11 or 14 amino acids; or

(b) a first ring-forming amino acid is linked to a second ring-forming amino acid to form a first ring comprising 4-11 or 14 amino acids; and a third ring-forming amino acid is linked to the C-terminus of the peptide to form a second ring comprising 4-11 or 14 amino acids.

Non-limiting examples of ring-forming amino acids include 4AminoPro, Abu, aG, aMeC, Api, C, D, Dap, Dap(N3), Dab, E, hA, hE, hK, K, Om, Pen, Pen(oXyl), Pen(mXyl), Pen(pXyl), Pra, R5H, R7H, R5Me, S5H, S7H, or S5Me. In some embodiments, the first ring comprises 4-9 or 11 amino acids. In some embodiments, the first ring comprises 4, 6, or 10 amino acids. In some embodiments, the first ring comprises 4 amino acids. In some embodiments, the first ring comprises 5 amino acids. In some embodiments, the first ring comprises 6 amino acids. In some embodiments, the first ring comprises 7 amino acids. In some embodiments, the first ring comprises 8 amino acids. In some embodiments, the first ring comprises 9 amino acids. In some embodiments, the first ring comprises 10 amino acids. In some embodiments, the first ring comprises 11 amino acids. In some embodiments, the first ring comprises 14 amino acids.

In some embodiments, the first ring comprises a linkage between two ring-forming amino acids having a structure selected from the following:

In some embodiments, the first ring is formed between X4 and X9, X4 and X13, X5 and X10, X3 and X13, or Xe and X9. In some embodiments, the first ring is formed between X4 and X9, X4 and X13, X5 and X10, X3 and X13, or Xe and X9 via a linker having one or more groups selected from the group consisting of a disulfide, thioether, amide, olefin, ether, alkylene, and triazole. In some embodiments, the first ring is formed between X4 and X9, X4 and X13, X5 and X10, X3 and X13, or Xe and X9 via a linker selected from the group consisting of a disulfide, thioether, amide, olefin, and triazole.

In some embodiments, the first ring is formed between X4 and X9, X4 and X13, or Xe and X9. In some embodiments, the first ring is formed between X4 and X9, X4 and X13, or Xe and X9 via a linker having one or more groups selected from the group consisting of a disulfide, thioether, amide, olefin, ether, alkylene, and triazole. In some embodiments, the first ring is formed between X4 and X9, X4 and X13, or Xe and X9 via a linker selected from the group consisting of a disulfide, thioether, amide, olefin, and triazole.

In some embodiments, the first ring is formed between X4 and X9. In some embodiments, the first ring is formed between X4 and X9 via a linker having one or more groups selected from the group consisting of a disulfide, thioether, amide, olefin, ether, alkylene, and triazole. In some embodiments, the first ring is formed between X4 and X9 via a linker selected from the group consisting of a disulfide, thioether, amide, olefin, and triazole.

In some embodiments, the first ring is formed between X4 and X13. In some embodiments, the first ring is formed between X4 and X13 via a linker having one or more groups selected from the group consisting of a disulfide, thioether, amide, olefin, ether, alkylene, and triazole. In some embodiments, the first ring is formed between X4 and X13 via a linker selected from the group consisting of a disulfide, thioether, amide, olefin, and triazole.

In some embodiments, the first ring is formed between X5 and X10. In some embodiments, the first ring is formed between X5 and X10 via a linker having one or more groups selected from the group consisting of a disulfide, thioether, amide, olefin, ether, alkylene, and triazole. In some embodiments, the first ring is formed between X5 and X10 via a linker selected from the group consisting of a disulfide, thioether, amide, olefin, and triazole.

In some embodiments, the first ring is formed between X3 and X13. In some embodiments, the first ring is formed between X3 and X13 via a linker having one or more groups selected from the group consisting of a disulfide, thioether, amide, olefin, ether, alkylene, and triazole. In some embodiments, the first ring is formed between X3 and X13 via a linker selected from the group consisting of a disulfide, thioether, amide, olefin, and triazole.

In some embodiments, the first ring is formed between Xe and X9. In some embodiments, the first ring is formed between Xe and X9 via a linker having one or more groups selected from the group consisting of a disulfide, thioether, amide, olefin, ether, alkylene, and triazole. In some embodiments, the first ring is formed between Xe and X9 via a linker selected from the group consisting of a disulfide, thioether, amide, olefin, and triazole.

In some embodiments, the second ring comprises 4-9 or 11 amino acids. In some embodiments, the second ring comprises 4, 6, 10 or 11 amino acids. In some embodiments, the second ring comprises 4 amino acids. In some embodiments, the second ring comprises 5 amino acids. In some embodiments, the second ring comprises 6 amino acids. In some embodiments, the second ring comprises 7 amino acids. In some embodiments, the second ring comprises 8 amino acids. In some embodiments, the second ring comprises 9 amino acids. In some embodiments, the second ring comprises 10 amino acids. In some embodiments, the second ring comprises 11 amino acids. In some embodiments, the second ring comprises 14 amino acids.

In some embodiments, the second ring comprises a linkage between two ring-forming

In some embodiments, the second ring comprises a linkage between the N-terminus of the peptide and a ring-forming amino acid and has a structure selected from the following:

In some embodiments, the second ring is formed between X4 and X9, X4 and X13, X5 and X10, X3 and X13, or Xe and X9. In some embodiments, the second ring is formed between X4 and X9, X4 and X13, X5 and X10, X3 and X13, or Xe and X9 via a linker having one or more groups selected from the group consisting of a disulfide, thioether, amide, olefin, ether, alkylene, and triazole. In some embodiments, the second ring is formed between X4 and X9, X4 and X13, X5 and X10, X3 and X13, or Xe and X9 via a linker selected from the group consisting of a disulfide, thioether, amide, olefin, and triazole.

In some embodiments, the second ring is formed between X5 and X10 or X3 and X13. In some embodiments, the second ring is formed between X5 and X10 or X3 and X13 via a linker having one or more groups selected from the group consisting of a disulfide, thioether, amide, olefin, ether, alkylene, and triazole. In some embodiments, the second ring is formed between X5 and X10 or X3 and X13 via a linker having one or more groups selected from the group consisting of a disulfide, thioether, amide, olefin, and triazole. In some embodiments, the second ring is formed between X5 and X10 or X3 and X13 via a linker selected from the group consisting of a disulfide, thioether, amide, olefin, and triazole.

In some embodiments, the second ring is formed between X4 and X9. In some embodiments, the second ring is formed between X4 and X9 via a linker having one or more groups selected from the group consisting of a disulfide, thioether, amide, olefin, ether, alkylene, and triazole. In some embodiments, the second ring is formed between X4 and X9 via a linker selected from the group consisting of a disulfide, thioether, amide, olefin, and triazole.

In some embodiments, the second ring is formed between X4 and X13. In some embodiments, the second ring is formed between X4 and X13 via a linker having one or more groups selected from the group consisting of a disulfide, thioether, amide, olefin, ether, alkylene, and triazole. In some embodiments, the second ring is formed between X4 and X13 via a linker selected from the group consisting of a disulfide, thioether, amide, olefin, and triazole.

In some embodiments, the second ring is formed between X5 and X10. In some embodiments, the second ring is formed between X5 and X10 via a linker having one or more groups selected from the group consisting of a disulfide, thioether, amide, olefin, ether, alkylene, and triazole. In some embodiments, the second ring is formed between X5 and X via a linker selected from the group consisting of a disulfide, thioether, amide, olefin, and triazole.

In some embodiments, the second ring is formed between X3 and X13. In some embodiments, the second ring is formed between X3 and X13 via a linker having one or more groups selected from the group consisting of a disulfide, thioether, amide, olefin, ether, alkylene, and triazole. In some embodiments, the second ring is formed between X3 and X13 via a linker selected from the group consisting of a disulfide, thioether, amide, olefin, and triazole.

In some embodiments, the second ring is formed between Xe and X9. In some embodiments, the second ring is formed between Xe and X9 via a linker having one or more groups selected from the group consisting of a disulfide, thioether, amide, olefin, ether, alkylene, and triazole. In some embodiments, the second ring is formed between Xe and X9 via a linker selected from the group consisting of a disulfide, thioether, amide, olefin, and triazole.

In some embodiments, the second ring is formed between X13 and the N-terminus of the peptide. In some embodiments, the second ring is formed between X13 and the N-terminus of the peptide via a linker having one or more groups selected from the group consisting of a disulfide, thioether, amide, olefin, ether, alkylene, and triazole. In some embodiments, the second ring is formed between X13 and the N-terminus of the peptide via a linker selected from the group consisting of a disulfide, thioether, amide, olefin, and triazole.

In some embodiments, the first ring is formed between X4 and X9 via a linker having one or more groups selected from the group consisting of a disulfide, thioether, amide, olefin, ether, alkylene, and triazole; and the second ring is formed between X3 and X13, between X5 and Xw, between Xw and X13, or between X13 and the N-terminus of the peptide via a linker having one or more groups selected from the group consisting of a disulfide, thioether, amide, olefin, ether, alkylene, and triazole.

In some embodiments, the first ring is formed between X4 and X9 via a linker selected from the group consisting of a disulfide, thioether, amide, olefin, and triazole; and the second ring is formed between X3 and X13, between X5 and Xw, between Xw and X13, or between X13 and the N-terminus of the peptide via a linker selected from the group consisting of a disulfide, thioether, amide, olefin, and triazole. In some embodiments, the first ring is formed between X4 and X13 via a linker having one or more groups selected from the group consisting of a disulfide, thioether, amide, olefin, ether, alkylene, and triazole; and the second ring is formed between X5 and X or between Xe and X9 via a linker having one or more groups selected from the group consisting of a disulfide, thioether, amide, olefin, ether, alkylene, and triazole.

In some embodiments, the first ring is formed between X4 and X13 via a linker selected from the group consisting of a disulfide, thioether, amide, olefin, and triazole; and the second ring is formed between X5 and X or between Xe and X9 via a linker selected from the group consisting of a disulfide, thioether, amide, olefin, and triazole.

In some embodiments, the first ring is formed between Xe and X9 via a linker having one or more groups selected from the group consisting of a disulfide, thioether, amide, olefin, ether, alkylene, and triazole; and the second ring is formed between X3 and X13, between X4 and X13, between X5 and Xw, between Xw and X13, or between X13 and the N-terminus of the peptide via a linker having one or more groups selected from the group consisting of a disulfide, thioether, amide, olefin, ether, alkylene, and triazole.

In some embodiments, the first ring is formed between Xe and X9 via a linker selected from the group consisting of a disulfide, thioether, amide, olefin, and triazole; and the second ring is formed between X3 and X13, between X4 and X13, between X5 and Xw, between Xw and X13, or between X13 and the N-terminus of the peptide via a linker selected from the group consisting of a disulfide, thioether, amide, olefin, and triazole.

In some embodiments, the peptide of Formula (I’) is a peptide of Formula (I).

Accordingly, in some embodiments, the present disclosure provides a peptide of Formula (I), comprising the amino acid sequence:

Ri-X3-X₄-X₅-T-X7-X8-X₉-Xw-2Nal-Xi2-Xi3-N-Xi₅-Xi6-R2 (I), or a pharmaceutically acceptable salt thereof, wherein:

Ri is MeCO, 8Aoc, 7Ahp, or cPEG3aCO;

X₃ is R7H, S7H, hK, or absent;

X4 is 4AminoPro, Abu, aG, aMeC, C, Dap, Pen, Pen(oXyl), Pen(mXyl), Pen(pXyl), or Pra;

X₅ is N, N(NMe2), Q, or Q(NMe2);

X₇ is 7MeW or W;

X₈ is K(Ac), R5H, S5H, K(NMeAc), or Q;

X₉ is aMeC, aG, C, D, E, hE, Pen, or Dap(N3);

Xw is AEF, APEG3F, F(4TzlAme2), or TMAPF;

Xi₂ is R5, S5, B5, or THP; X13 is E, R5H, or S5H;

X15 is 3Pya or bAla;

Xi6 is R5H, S5H, Sar, or absent;

R₂ is CONH₂ or CONMe2; wherein when X3 is R7H or S7H, then X5 is N(Me2) or Q(NMe2), Xs is K(NmeAc), or X10 is APEG3F; wherein the peptide is cyclized via:

(a) a first linkage between the residues at X4 and X9; and

(b) a second linkage selected from the group consisting of a linkage between 8Aoc at Ri and E at X13, a linkage between 7Ahp at Ri and E at X13, a linkage between R7H or S7H at X3 and R5H or S5H at X13, a linkage between hK at X3 and E at X13, a linkage between R5H or S5H at Xs and R5 or S5 at X12, a linkage between F(4TzlAme2) at X10 and E at X13, a linkage between AEF at X10 and E at X13; a linkage between R5 or S5 at X12 and R5H or S5H at Xie; and two linkages between B5 at X12 and R5H or S5H at Xs and between B5 at X12 and R5H or S5H at Xie.

Ri-X3-X4-X₅-T-X7-X8-X₉-Xio-2Nal-Xi2-Xi3-N-Xi₅-Xi6-R2 (I), or a pharmaceutically acceptable salt thereof, wherein:

Ri is MeCO, 8Aoc, 7Ahp, or cPEG3aCO;

X₃ is R7H, S7H, hK, or absent;

X₄ is 4AminoPro, Abu, aG, aMeC, C, Dap, Pen, Pen(oXyl), Pen(mXyl), Pen(pXyl), or Pra;

X₅ is N, N(NMe2), Q, or Q(NMe2);

X₇ is 7MeW or W;

X₈ is K(Ac), R5H, S5H, K(NMeAc), or Q;

X₉ is aMeC, aG, C, D, E, hE, Pen, or Dap(N3);

X10 is AEF, APEG3F, F(4TzlAme2), or TMAPF;

X₁₂ is R5, S5, B5, or THP;

X13 is E, R5H, or S5H;

X15 is 3Pya or bAla; Xi6 is R5H, S5H, Sar, or absent;

(c) a first linkage between the residues at X4 and X9; and

(d) a second linkage selected from the group consisting of a linkage between 8Aoc at Ri and E at X13, a linkage between R7H or S7H at X3 and R5H or S5H at X13, a linkage between hK at X3 and E at X13, a linkage between R5H or S5H at Xs and R5 or S5 at X12, a linkage between F(4TzlAme2) at X10 and E at X13, a linkage between AEF at X10 and E at X13; a linkage between R5 or S5 at X12 and R5H or S5H at Xie; and two linkages between B5 at X12 and R5H or S5H at Xs and between B5 at X12 and R5H or S5H at Xie.

In some embodiments, the peptide comprises an amino acid sequence of Formula (I-A):

Ri-X3-X₄-X₅-T-X7-X8-X₉-Xio-2Nal-Xi2-Xi3-N-3Pya-Xi6-R2 (I-A) or a pharmaceutically acceptable salt thereof, wherein:

Ri is MeCO, 8Aoc, or cPEG3aCO;

X₃ is R7H, S7H, hK, or absent;

X₅ is N or N(NMe₂);

X₇ is 7MeW or W;

X₈ is K(Ac), R5H, S5H, K(NMeAc), or Q;

X₉ is aMeC, aG, C, D, E, hE, Pen, or Dap(N3);

X10 is AEF, APEG3F, F(4TzlAme2), or TMAPF;

X₁₂ is R5, S5, B5, or THP;

X13 is E, R5H, or S5H;

X15 is 3Pya or bAla;

Xi6 is S5H or Sar; and

R₂ is CONH2 or CONMe2; wherein when X3 is R7H or S7H, then X5 is N(Me2), X₈ is K(NmeAc), or X10 is APEG3F; wherein the peptide is cyclized via:

(e) a first linkage between the residues at X4 and X9; and

(f) a second linkage selected from the group consisting of a linkage between 8Aoc at Ri and E at X13, a linkage between R7H or S7H at X3 and R5H or S5H at X13, a linkage between hK at X3 and E at X13, a linkage between R5H or S5H at Xs and R5 or S5 at X12, a linkage between F(4TzlAme2) at X10 and E at X13, a linkage between AEF at X10 and E at X13; a linkage between R5 or S5 at X12 and R5H or S5H at Xie; and two linkages between B5 at X12 and R5H or S5H at Xs and between B5 at X12 and R5H or S5H at Xie.

In some embodiments, the peptide comprises an amino acid sequence of Formula (I-B):

Ri-X3-Pen-X₅-T-7MeW-Xs-Pen-Xio-2Nal-Xi2-Xi3-N-3Pya-Xi6-R2 (I-B) or a pharmaceutically acceptable salt thereof, wherein:

Ri is MeCO, 8Aoc, or cPEG3aCO;

X₃ is R7H, S7H, hK, or absent;

X₅ is N or N(NMe₂);

X₈ is K(Ac), S5H, or K(NMeAc);

X10 is AEF, APEG3F, F(4TzlAme2), or TMAPF;

X₁₂ is R5, S5, B5, or THP;

X13 is E, R5H, or S5H;

Xie is S5H or Sar; and

R₂ is CONH2 or CONMe2; wherein when X3 is R7H or S7H, then X5 is N(Me2) or Q(NMe2), X₈ is K(NmeAc), or X10 is APEG3F; wherein the peptide is cyclized via:

(g) a first linkage between the residues at X4 and X9; and

(h) a second linkage selected from the group consisting of: a linkage between 8Aoc at Ri and E at X13, a linkage between R7H or S7H at X3 and R5H or S5H at X13, a linkage between hK at X3 and E at X13, a linkage between R5H or S5H at X₈ and R5 or S5 at X12, a linkage between F(4TzlAme2) at X10 and E at X13, a linkage between AEF at X10 and E at X13; a linkage between R5 or S5 at X12 and R5H or S5H at Xie; and two linkages between B5 at X12 and R5H or S5H at Xs and between B5 at X12 and R5H or S5H at Xie.

In some embodiments, the peptide comprises an amino acid sequence of Formula (I-C):

Ri-X3-X₄-X₅-T-X7-X8-X₉-Xio-2Nal-THP-Xi3-N-3Pya-Xi6-R2 (I-C) or a pharmaceutically acceptable salt thereof, wherein:

Ri is MeCO, 8Aoc, 7Ahp, or cPEG3aCO;

X₃ is R7H, S7H, hK, or absent;

X₅ is N, N(NMe2), Q, or Q(NMe2);

X₇ is 7MeW or W;

X₈ is K(Ac), K(NMeAc), or Q;

X₉ is aMeC, aG, C, D, E, hE, Pen, or Dap(N3);

X10 is AEF, APEG3F, F(4TzlAme2), or TMAPF;

X13 is E, R5H, or S5H;

Xi6 is Sar or absent; and

(i) a first linkage between the residues at X₄ and X₉; and

(j) a second linkage selected from the group consisting of: a linkage between 8Aoc at Ri and E at X13, a linkage between R7H or S7H at X3 and R5H or S5H at X13, a linkage between hK at X3 and E at X13, a linkage between R5H or S5H at X₈ and R5 or S5 at X12, a linkage between F(4TzlAme2) at X10 and E at X13, a linkage between AEF at X10 and E at X13;

In some embodiments, the peptide comprises an amino acid sequence of Formula (I-D):

Ri-X3-X₄-X₅-T-X₇-X₈-X₉-Xio-2Nal-Xi2-E-N-3Pya-Xi6-CONH2 (I-D) wherein:

Ri is MeCO or 8Aoc;

X3 is absent; X4 is 4AminoPro, Abu, aG, aMeC, C, Dap, Pen, Pen(oXyl), Pen(mXyl), Pen(pXyl), or

Pra;

X5 is N or Q;

X₇ is 7MeW or W;

X₈ is K(Ac), S5H, or Q;

X₉ is aMeC, aG, C, D, E, hE, Pen, or Dap(N3);

X10 is AEF or F(4TzlAme2);

X12 is R5, S5, B5, or THP; and

Xi6 is S5H or Sar; wherein the peptide is cyclized via:

(k) a first linkage between the residues at X4 and X₉; and

(l) a second linkage selected from the group consisting of a linkage between 8Aoc at Ri and E at X13, a linkage between R5H or S5H at X₈ and R5 or S5 at X12, a linkage between F(4TzlAme2) at X10 and E at X13, a linkage between AEF at X10 and E at X13; a linkage between R5 or S5 at X12 and R5H or S5H at Xie; and two linkages between B5 at X12 and R5H or S5H at X₈ and between B5 at X12 and R5H or S5H at Xie.

In some embodiments, Ri is 8Aoc, 7Ahp, cPEG3aCO, or MeOC, wherein the 8Aoc is linked to the amino acid at X13. In some embodiments, Ri is 8Aoc, 7Ahp, cPEG3aCO, or MeCO, wherein the 8Aoc is linked to an E residue at X13 via an amino linkage.

In some embodiments, Ri is 7Ahp, 8Aoc, or MeCO. In some embodiments, Ri is 7Ahp, cPEG3aCO, or MeCO. In some embodiments, Ri is 8Aoc, cPEG3aCO, or MeCO. In some embodiments, Ri is MeCO or 7Ahp. In some embodiments, Ri is 8Aoc or MeCO. In some embodiments, Ri is MeCO or cPEG3aCO.

In some embodiments, Ri is an alkyl chain linked to the amino acid at X13. In some embodiments, Ri is 8Aoc linked to the amino acid at X13. In some embodiments, Xi is 8Aoc linked to E at X13. In some embodiments, Ri is 7Ahp. In some embodiments, Ri is 7Ahp linked to the amino acid at X13. In some embodiments, Ri is 7Ahp linked to E at X13. In some embodiments, Ri is MeCO. In some embodiments, Ri is cPEG3aCO.

In some embodiments, X3 is hK, R7H, S7H, or absent; wherein the hK, R7H, and S7H are linked to the amino acid at X13. In some embodiments, X3 is hK, R7H, S7H, or absent; wherein the hK, R7H, and S7H are linked to the amino acid at X13, and wherein the hK is an L amino acid. In some embodiments, X3 is hk, R7H, S7H, or absent; wherein the hk, R7H, and S7H are linked to the amino acid at X13. In some embodiments, X3 is hK, R7H, S7H, or absent. In some embodiments, X3 is hk, R7H, S7H, or absent.

In some embodiments, X3 is hk, R7H, S7H, or absent; wherein the hk, R7H, and S7H are linked to an R5H, S5H, or E residue at X13. In some embodiments, X3 is hk, R7H, S7H, or absent; wherein the hk, R7H, and S7H are linked to an amino acid at X13 via an aliphatic or amide linkage. In some embodiments, X3 is hk, R7H, S7H, or absent; wherein the R7H and S7H are linked to an R5H or S5H residue at X13 via an aliphatic linkage. In some embodiments, X3 is hk, R7H, S7H, or absent; wherein the hk is linked to an E residue at X13 via an amide linkage.

In some embodiments, X3 is R7H, hk or absent. In some embodiments, X3 is hk or absent.

In some embodiments, X3 is R7H linked to the amino acid at X13. In some embodiments, X3 is R7H linked to R5H at X13. In some embodiments, X3 is S7H linked to the amino acid at X13. In some embodiments, X3 is S7H linked to S5H at X13. In some embodiments, X3 is hk linked to the amino acid at X13. In some embodiments, X3 is hk linked to E at X13. In some embodiments, X3 is hK linked to the amino acid at X13. In some embodiments, X3 is hK linked to E at X13. In some embodiments, X3 is absent.

In some embodiments, X4 is 4AminoPro, Abu, aG, aMeC, C, Dap, Pen, Pen(oXyl), Pen(mXyl), Pen(pXyl), or Pra. In some embodiments, X4 is 4AminoPro, Abu, aG, aMeC, C, Dap, Pen, Pen(oXyl), Pen(mXyl), Pen(pXyl), or Pra, each of which is an L amino acid. In some embodiments, X4 is 4Amino-d-Pro, dAbu, d-aG, aMe-d-C, c, dDap, dPen, dPen(oXyl), dPen(mXyl), dPen(pXyl), or dPra.

In some embodiments, X4 is 4AminoPro, aG, Dap, Pen(oXyl), Pen(mXyl), Pen(pXyl), or Pra. In some embodiments, X4 is Abu, aMeC, C, Pen, Pen(oXyl), Pen(mXyl), or Pen(pXyl). In some embodiments, X4 is Abu, aMeC, C, or Pen. In some embodiments, X4 is Abu, C, or Pen. In some embodiments, X4 is Abu or Pen.

In some embodiments, X4 is 4AminoPro. In some embodiments, X4 is 4RAminoPro. In some embodiments, X4 is 4SAminoPro. In some embodiments, X4 is Abu. In some embodiments, X4 is aG. In some embodiments, X4 is aMeC. In some embodiments, X4 is C. In some embodiments, X4 is Dap. In some embodiments, X4 is Pen. In some embodiments, X4 is Pen(oXyl). In some embodiments, X4 is Pen(mXyl). In some embodiments, X4 is Pen(pXyl). In some embodiments, X4 is Pra.

In some embodiments, X4 is 4Amino-d-Pro. In some embodiments, X4 is 4RAmino-d- Pro. In some embodiments, X4 is 4SAmino-d-Pro. In some embodiments, X4 is dAbu. In some embodiments, X4 is d-aG. In some embodiments, X4 is aMe-d-C. In some embodiments, X4 is c. In some embodiments, X4 is dDap. In some embodiments, X4 is dPen. In some embodiments, X4 is dPen(oXyl). In some embodiments, X4 is dPen(mXyl). In some embodiments, X4 is dPen(pXyl). In some embodiments, X4 is dPra.

In some embodiments, X5 is N, N(NMe2), Q, or Q(NMe2). In some embodiments, X5 is N, N(NMe2), Q, or Q(NMe2), each of which is an L amino acid. In some embodiments, X5 n, n(NMe2), q, or q(NMe2).

In some embodiments, X5 is N, N(NMe2) or Q. In some embodiments, X5 is N, Q, or Q(NMe2). In some embodiments, X5 is N or Q. In some embodiments, X5 is N. In some embodiments, when X5 is N, then X4 is Pen. In some embodiments, X5 is Q. In some embodiments, when X5 is Q, then X4 is Abu.

In some embodiments, Xe is T. In some embodiments, Xe is T, wherein the T is an L- amino acid. In some embodiments, Xe is t.

In some embodiments, X7 is 7MeW or W. In some embodiments, X7 is 7MeW or W, each of which is an L-amino acid. In some embodiments, X7 is 7Mew or w.

In some embodiments, X7 is 7MeW. In some embodiments, X7 is 7Mew. In some embodiments, X?is W. In some embodiments, X7 is w. In some embodiments, when X7 is W, then X5 is Q. In some embodiments, when X7 is W, then X4 is Abu. In some embodiments, when X7 is W, then X5 is Q and X4 is Abu.

In some embodiments, Xs is K(Ac), R5H, S5H, K(NMeAc), or Q. In some embodiments, Xs is K(Ac), R5H, S5H, K(NMeAc), or Q, wherein Q, K(Ac), and K(NMeAc) are L-amino acids. In some embodiments, Xs is k(Ac), R5H, S5H, k(NMeAc), or q.

In some embodiments, Xs is K(Ac), R5H, S5H, K(NMeAc), or Q, wherein the R5H and S5H are linked to an amino acid at X12. In some embodiments, Xs is K(Ac), R5H, S5H, K(NMeAc), or Q, wherein the R5H and S5H are linked to an amino acid at X12 via an aliphatic linkage.

In some embodiments, Xs is K(Ac), S5H, K(NMeAc), or Q. In some embodiments, Xs is K(Ac), S5H, or Q. In some embodiments, Xs is K(Ac), K(NMeAc), or Q. In some embodiments, Xs is K(Ac) or Q. In some embodiments, Xs is K(Ac) or K(NMeAc). In some embodiments, Xs is K(Ac). In some embodiments, Xs is K(NMeAc). In some embodiments, Xs is Q. In some embodiments, Xs is k(Ac). In some embodiments, Xs is k(NMeAc). In some embodiments, Xs is q-

In some embodiments, Xs is R5H or S5H. In some embodiments, Xs is S5H linked to S5 at X12. In some embodiments, Xs is R5H linked to R5 at X12. In some embodiments, Xs is S5H linked to R5 at X12. In some embodiments, Xs is R5H linked to S5 at X12. In some embodiments, X9 is aMeC, aG, C, D, E, hE, Pen, or Dap(N3). In some embodiments, X9 is aMeC, aG, C, D, E, hE, Pen, or Dap(N3), each of which is an L-amino acid. In some embodiments, X9 is aMe-d-C, d-aG, c, d, e, he, dPen, or dDap(N3).

In some embodiments, X9 is aG, D, E, hE, or Dap(N3). In some embodiments, X9 is aMeC, C, or Pen. In some embodiments, X9 is aMeC or Pen.

In some embodiments, X9 is aMeC. In some embodiments, X9 is aG. In some embodiments, X9 is C. In some embodiments, X9 is D. In some embodiments, X9 is E. In some embodiments, X9 is hE. In some embodiments, X9 is Pen. In some embodiments, X9 is Dap(N3).

In some embodiments, X9 is aMe-d-C. In some embodiments, X9 is d-aG. In some embodiments, X9 is c. In some embodiments, X9 is d. In some embodiments, X9 is e. In some embodiments, X9 is he. In some embodiments, X9 is dPen. In some embodiments, X9 is dDap(N3).

In some embodiments, X10 is AEF, APEG3F, F(4TzlAme2), or TMAPF, optionally wherein the AEF is linked to the amino acid at X13. In some embodiments, X10 is AEF, APEG3F, F(4TzlAme2), or TMAPF, each of which is an L-amino acid, optionally wherein the AEF is linked to the amino acid at X13. In some embodiments, X10 is dAEF, dAPEG3F, f(4TzlAme2), or dTMAPF, optionally wherein the dAEF is linked to the amino acid at X13.

In some embodiments, X10 is AEF, APEG3F, F(4TzlAme2), or TMAPF. In some embodiments, X10 is AEF, TMAPF, or APEG3F. In some embodiments, X10 is AEF, TMAPF, or F(4TzlAme2). In some embodiments, X10 is AEF or TMAPF.

In some embodiments, X10 is APEG3F. In some embodiments, X10 is F(4TzlAme). In some embodiments, X10 TMAPF. In some embodiments, X10 is AEF. In some embodiments, X10 is AEF linked to the amino acid at X13. In some embodiments, X10 is AEF linked to E at X13. In some embodiments, X10 is dAPEG3F. In some embodiments, X10 is f(4TzlAme). In some embodiments, X10 dTMAPF. In some embodiments, X10 is dAEF. In some embodiments, X10 is dAEF linked to the amino acid at X13. In some embodiments, X10 is dAEF linked to E at X13.

In some embodiments, X12 is THP, S5, R5, or B5. In some embodiments, X12 is THP, S5, R5, or B5, wherein the S5 and R5 are linked to an amino acid at Xi6. In some embodiments, X12 is THP, S5, R5, or B5, wherein the B5 is linked to an amino acid at Xs and an amino acid at Xi6. In some embodiments, X12 is THP, S5, or R5. In some embodiments, X12 is THP, S5, or B5. In some embodiments, X12 is THP or S5. In some embodiments, X12 is THP or B5. In some embodiments, X12 is THP. In some embodiments, X12 is S5. In some embodiments, X12 is R5. In some embodiments, X12 is B5.

In some embodiments, X12 is S5 linked to an amino acid at Xi6 via an aliphatic linkage. In some embodiments, X12 is THP, S5, R5, or B5, wherein the S5 and R5 are linked to S5H or R5H at Xi6. In some embodiments, X12 is S5 linked to S5H at Xi6. In some embodiments, X12 is R5 linked to an amino acid at Xi6 via an aliphatic linkage. In some embodiments, X12 is R5 linked to R5H at Xi6.

In some embodiments, X12 is B5 linked to an amino acid at Xs and an amino acid at Xi6. In some embodiments, X12 is B5 linked to an amino acid at Xs via a first aliphatic linkage and an amino acid at Xi6 via a second aliphatic linkage. In some embodiments, X12 is B5 linked to S5H at Xs and S5H at Xi6. In some embodiments, X12 is B5 linked to R5H at Xs and R5H at Xi6.

In some embodiments, X13 is E, R5H, or S5H; wherein R5H, and S5H are linked to Ri, the amino acid at X3 and optionally wherein the E is linked to Ri, the amino acid at X3, or the amino acid at X10. In some embodiments, X13 is E, R5, or S5H, wherein the E is an L-amino acid; wherein R5H, and S5H are linked to the amino acid at X3 and optionally wherein the E is linked to Ri, the amino acid at X3, or the amino acid at X10. In some embodiments, X13 is e, R5H, S5H; wherein the R5H and S5H are linked to the amino acid at X3, and optionally wherein the e is linked to Ri, the amino acid at X3, or the amino acid at X10.

In some embodiments, X13 is E, R5H, or S5H. In some embodiments, X13 is E or S5H. In some embodiments, X13 is E or R5H. In some embodiments, X13 is E.

In some embodiments, X13 is E linked to Ri. In some embodiments, X13 is E linked to Ri via an amide linkage. In some embodiments, X13 is E linked to 8Aoc at Ri. In some embodiments, X13 is E linked to the amino acid at X3. In some embodiments, X13 is E linked to the amino acid at X3 via an amide linkage. In some embodiments, X13 is E linked to hK at X3. In some embodiments, X13 is E linked to the amino acid at X10. In some embodiments, X13 is E linked to the amino acid at X10. In some embodiments, X13 is E linked to the amino acid at X10 via an amide linkage. In some embodiments, X13 is E linked to AEF at X10. In some embodiments, X13 is E linked to the amino acid at X10 via a tetrazolyl linkage. In some embodiments, X13 is E linked to F(4TzlAme2) at X10. In some embodiments, X13 is S5H. In some embodiments, X13 is S5H linked to X3. In some embodiments, X13 is S5H linked to R7H at X₃.

In some embodiments, X14 is N. In some embodiments, X14 is N, wherein the N is an L- amino acid. In some embodiments, X14 is n.

In some embodiments, X15 is 3Pya or bAla. In some embodiments, X15 is 3Pya or bAla, wherein the 3Pya is an L amino acid. In some embodiments, X15 is d-3Pya or bAla.

In some embodiments, X15 is 3Pya. In some embodiments, X15 is bAla. In some embodiments, X15 is d-3Pya.

In some embodiments, Xi6 is Sar, R5H, S5H, or absent, wherein the S5H and R5H are linked to an amino acid at X12. In some embodiments, Xi6 is Sar, R5h, S5H, or absent, wherein the S5H and R5H are linked to an amino acid at X12 via an aliphatic linkage. In some embodiments, Xi6 is Sar, S5H, R5H or absent, wherein the S5H and R5H are linked to B5 at X12. In some embodiments, Xi6 is Sar, S5H, or absent. In some embodiments, Xi6 is Sar, R5H, or absent.

In some embodiments, Xi6 is Sar or absent. In some embodiments, Xi6 is Sar or S5H. In some embodiments, Xi6 is Sar or R5H. In some embodiments, Xi6 is S5H. In some embodiments, Xi6 is R5H. In some embodiments, Xi6 is R5H linked to B5 at X12. In some embodiments, Xi6 is S5H linked to an amino acid at X12. In some embodiments, Xi6 is S5H linked to B5 at X12. In some embodiments, Xi6 is Sar. In some embodiments, Xi6 is absent.

In some embodiments, R2 is CONH2 or CONMe2. In some embodiments, R2 is CONH2. In some embodiments, R2 is CON(Me)2.

In some embodiments, the peptide is cyclized via a linkage between two amino acid residues (e.g., the residues of X4 and X9) via a disulfide, thioether, amide, or alkylene bond.

In some embodiments, X4 is Abu and X9 is C. In some embodiments, X4 is Abu and X9 is aMeC. In some embodiments, X4 is Abu and X9 is Pen.

In some embodiments, X4 is C and X9 is C. In some embodiments, X4 is C and X9 is aMeC. In some embodiments, X4 is C and X9 is Pen.

In some embodiments, X4 is Pen and X9 is C. In some embodiments, X4 is Pen and X9 is aMeC. In some embodiments, X4 is Pen and X9 is Pen.

In some embodiments, X4 is aMeC and X9 is C. In some embodiments, X4 is aMeC and X9 is aMeC. In some embodiments, X4 is aMeC and X9 is Pen.

In some embodiments, X4 is Pen(oXyl) and X9 is C. In some embodiments, X4 is Pen(oXyl) and X9 is aMeC. In some embodiments, X4 is Pen(oXyl) and X9 is Pen.

In some embodiments, X4 is Pen(mXyl) and X9 is C. In some embodiments, X4 is Pen(mXyl) and X9 is aMeC. In some embodiments, X4 is Pen(mXyl) and X9 is Pen.

In some embodiments, X4 is Pen(pXyl) and X9 is C. In some embodiments, X4 is Pen(pXyl) and X9 is aMeC. In some embodiments, X4 is Pen(pXyl) and X9 is Pen.

In some embodiments, X4 is 4AminoPro and X9 is D. In some embodiments, X4 is 4AminoPro and X9 is E. In some embodiments, X4 is 4AminoPro and X9 is hE.

In some embodiments, X4 is Dap and X9 is D. In some embodiments, X4 is Dap and X9 is E. In some embodiments, X4 is Dap and X9 is hE.

In some embodiments, X4 is Pra and X9 is Dap(N3). In some embodiments, X4 is aG and X9 is aG.

In some embodiments, the peptide is cyclized via a linkage between two amino acid residues (e.g., at X4 and X9) having a structure selected from the following: In some embodiments, the peptide is cyclized via a linkage between the residues at X4 and X9 having a structure selected from the following:

In some embodiments, the peptide is cyclized via a linkage between the residues at X4 and X9 having the following structure:

Pen - Pen.

In some embodiments, the peptide comprises a linkage between Ri and X13 having a

In some embodiments, the peptide comprises a linkage between X3 and X13 having a structure selected from the following:

In some embodiments, the peptide comprises a linkage between Xs and X12 having the In some embodiments, the peptide comprises a linkage between X10 and X13 having a

In some embodiments, the peptide comprises a linkage between X12 and Xi6 having the following structure:

In some embodiments, the peptide comprises two linkages, one between X12 and Xs, and one between X12 and Xi6, having the following structure:

In some embodiments, the peptide comprises a sequence according to any one of the following Formulas:

Ri-X3-Pen-X₅-T-7MeW-X₈-Pen-Xio-2Nal-Xi2-E-N-3Pya-Xi6-R2 (I-E)

Ri-X3-X₄-X₅-T-X₇-X₈-X9-AEF-2Nal-THP-Xi3-N-3Pya-R2 (I-F)

Ri-X3-X₄-X₅-T-X7-X₈-X₉-AEF-2Nal-Xi2-E-N-3Pya-CONH2 (I- J)

MeCO-X3-X₄-X₅-T-X₇-X₈-X₉-AEF-2Nal-THP-E-N-3Pya-R2 (I-K)

MeCO-X₃-X₄-X₅-T-X₇-X₈-X₉-AEF-2Nal-THP-E-N-3Pya-CONH2 (I-L)

Ri-X3-Pen-X₅-T-7MeW-X₈-Pen-Xio-2Nal-Xi2-E-N-Xi₅-Xi6-R2 (I-M)

Ri-X3-Abu-Q-T-X₇-X₈-C-Xio-2Nal-Xi2-E-N-3Pya-Xi6-R2 (I-N)

Ri-X₃-Abu-Q-T-X₇-Q-C-Xio-2Nal-Xi2-E-N-3Pya-Xi6-R2 (I-O)

MeCO-X₃-X₄-K(Ac)-T-7MeW-X₈-X₉-Xio-2Nal-Xi2-E-N-3Pya-Sar-R2 (I-P)

Ri-X3-X₄-X₅-T-X₇-S5H-X₉-Xio-2Nal-B5-E-N-3Pya-S5H-R2 (I-Q)

Ri-X3-X₄-X₅-T-7MeW-S5H-X₉-Xio-2Nal-B5-E-N-3Pya-S5H-R2 (I-R)

Ri-X3-Pen-X₅-T-X₇-S5H-Pen-Xio-2Nal-B5-E-N-3Pya-S5H-R2 (I-S)

In some embodiments, the amino acid at X3 is an L-amino acid. In some embodiments, the amino acid at X3 is a D-amino acid.

In some embodiments, the amino acid at X₄ is an L-amino acid. In some embodiments, the amino acid at X₄ is a D-amino acid.

In some embodiments, the amino acid at X5 is an L-amino acid. In some embodiments, the amino acid at X5 is a D-amino acid.

In some embodiments, the T between X5 and X₇ is an L-amino acid. In some embodiments, the T between X5 and X₇ is dT.

In some embodiments, the amino acid at X₇ is an L-amino acid. In some embodiments, the amino acid at X₇ is a D-amino acid.

In some embodiments, the amino acid at X₈ is an L-amino acid. In some embodiments, the amino acid at X₈ is a D-amino acid. In some embodiments, the amino acid at X9 is an L-amino acid. In some embodiments, the amino acid at X9 is a D-amino acid.

In some embodiments, the amino acid at X10 is an L-amino acid. In some embodiments, the amino acid at X10 is a D-amino acid. In some embodiments, the 2Nal between Xw and X12 is an L-amino acid. In some embodiments, the 2Nal between Xw and X12 is a d2Nal.

In some embodiments, the amino acid at X13 is an L-amino acid. In some embodiments, the amino acid at X13 is a D-amino acid.

In some embodiments, the N between X13 and X15 is an L-amino acid. In some embodiments, the N between X13 and X15 is dN.

In some embodiments, the amino acid at X15 is an L-amino acid. In some embodiments, the amino acid at X15 is a D-amino acid.

In some embodiments, the present disclosure provides a peptide described herein provided the peptide retains activity as an inhibitor of interleukin-23 receptor. The present disclosure further provides a peptide of any one of SEQ ID NOS: 1-8, as shown in Table 2, or a pharmaceutically acceptable salt thereof.

Table 2. Example Peptides

In the peptide sequences shown above, where a number in parenthesis follows a particular residue, that residue is linked to another residue in the sequence that is denoted with the same number. For example, in the sequence MeCO-Pen(3)-N-T-7MeW-K(Ac)-Pen(3)-AEF- 2Nal-S5(4)-E-N-3Pya-S5H(4)-CONH2 (SEQ ID NO: 1), the two Pen(3) residues are linked to one another, and the S5(4) residue is linked to the S5H(4) residue.

Methods of Synthesis

The compounds described herein may be synthesized by many techniques that are known to those skilled in the art. In some aspects, the present disclosure provides a method of chemically synthesizing a peptide of the present disclosure. In some embodiments, a portion of the peptide is recombinantly synthesized, instead of being chemically synthesized. In some aspects, methods of producing a peptide further include cyclizing the peptide precursor after the constituent subunits have been attached. In particular aspects, cyclization is accomplished via any of the various methods described herein.

The present disclosure further describes synthesis of compounds described herein. In some aspects, one or more of the amino acid residues or amino acid monomers are lipidated and then covalently attached to one another to form a peptide of the disclosure. In some aspects, one or more of the amino acid residues or amino acid monomers are covalently attached to one another and lipidated at an intermediate oligomer stage before attaching additional amino acids and cyclization to form a peptide of the disclosure. In some aspects, a cyclic peptide is synthesized and then lipidated to form a compound of the disclosure. Illustrative synthetic methods are described in the Examples.

Pharmaceutical Compositions

The present disclosure further relates to a pharmaceutical composition comprising an IL-23R inhibitor described herein. In particular, the present disclosure includes pharmaceutical compositions comprising one or more peptides of the present disclosure and a pharmaceutically acceptable carrier, diluent or excipient. The pharmaceutically acceptable carrier, diluent or excipient may be a solid, semi-solid or liquid filler, diluent, encapsulating material or formulation auxiliary of any type. Prevention of the action of microorganisms may be ensured by the inclusion of various antibacterial and antifungal agents, for example, paraben, chlorobutanol, phenol sorbic acid, and the like. It may also be desirable to include isotonic agents such as sugars, sodium chloride, and the like.

The pharmaceutical compositions may be administered orally, parenterally, intraci stemally, intravaginally, intraperitoneally, intrarectally, topically (as by powders, ointments, drops, suppository, or transdermal patch), by inhalation (such as intranasal spray), ocularly (such as intraocularly) or buccally. The term “parenteral” as used herein refers to modes of administration which include intravenous, intramuscular, intraperitoneal, intrastemal, subcutaneous, intradermal and intraarticular injection and infusion. Accordingly, in certain embodiments, the compositions are formulated for delivery by any of these routes of administration. A pharmaceutical composition may be formulated for and administered orally. A pharmaceutical composition may be formulated for and administered parenterally.

The IL-23R inhibitors of the present disclosure may be prepared and/or formulated as pharmaceutically acceptable salts and/or other forms thereof or when appropriate in neutral form. Pharmaceutically acceptable salts are non-toxic salts of a neutral form of a compound that possess the desired pharmacological activity of the neutral form. These salts may be derived from inorganic or organic acids or bases. For example, a compound that contains a basic nitrogen may be prepared as a pharmaceutically acceptable salt by contacting the compound with an inorganic or organic acid. Non-limiting examples of pharmaceutically acceptable salts can be found in Remington: The Science and Practice of Pharmacy, 21^st Edition, Lippincott Wiliams and Wilkins, Philadelphia, Pa., 2006. The present disclosure relates to pharmaceutical compositions comprising an IL-23R inhibitor described herein or pharmaceutically acceptable salts, isomers, or a mixture thereof, in which one or more hydrogen atoms attached to a carbon atom may be replaced by a deuterium atom or D. As known in the art, the deuterium atom is a non-radioactive isotope of the hydrogen atom. Such compounds may increase resistance to metabolism, and thus may be useful for increasing the half-life of the compounds described herein or pharmaceutically acceptable salts, isomer, or a mixture thereof when administered to a mammal. See, e.g., Foster, “Deuterium Isotope Effects in Studies of Drug Metabolism,” Trends Pharmacol. Sci., 5(12):524-527 (1984). Such compounds are synthesized by means well known in the art, for example by employing starting materials in which one or more hydrogen atoms have been replaced by deuterium.

Examples of isotopes that can be incorporated into the disclosed compounds also include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorous, fluorine, chlorine, and iodine, such as ²H, ³H, ⁿC, ¹³C, ¹⁴C, ¹³N, ¹⁵N, ¹⁵0, ¹⁷O, ¹⁸O, ³¹P, ³²P, ³⁵S, ¹⁸F, ³⁶C1, ¹²³I, and ¹²⁵I, respectively. Substitution with positron emitting isotopes, such as ⁿC, ¹⁸F, ¹⁵O and ¹³N, can be useful in Positron Emission Topography (PET) studies for examining substrate receptor occupancy. Isotopically-labeled peptides of the present disclosure can generally be prepared by conventional techniques known to those skilled in the art or by processes analogous to those described in the Examples as set out below using an appropriate isotopically-labeled reagent in place of the non-labeled reagent previously employed.

When used in at least one of the treatments or delivery systems described herein, a peptide inhibitor of the present disclosure may be employed in pure form or, where such forms exist, in pharmaceutically acceptable salt form.

The total daily usage of the IL-23R inhibitor and compositions of the present disclosure can be decided by the attending physician within the scope of sound medical judgment. The specific therapeutically effective dose level for any particular subject will depend upon a variety of factors including: a) the disorder being treated and the severity of the disorder; b) activity of the specific compound employed; c) the specific composition employed, the age, body weight, general health, sex and diet of the patient; d) the time of administration, route of administration, and rate of excretion of the specific peptide inhibitor employed; e) the duration of the treatment; f) drugs used in combination or coincidental with the specific peptide inhibitor employed, and like factors well known in the medical arts.

The compositions may conveniently be presented in unit dosage form and can be prepared by any of the methods well known in the art of pharmacy. Techniques and compositions generally are found in Remington's Pharmaceutical Sciences (Mack Publishing Co., Easton, PA). Such methods include the step of bringing into association the active ingredient with the carrier which constitutes one or more accessory ingredients. In general the compositions are prepared by uniformly and intimately bringing into association the active ingredient with liquid carriers or finely divided solid carriers or both, and then, if necessary, shaping the product.

Non-Invasive Detection of Intestinal Inflammation

The IL-23R inhibitors of the present disclosure may be used for detection, assessment and diagnosis of intestinal inflammation by microPET imaging, wherein the peptide inhibitor is labeled with a chelating group or a detectable label, as part of a non-invasive diagnostic procedure. In certain embodiments, an IL-23R inhibitor of the present disclosure is conjugated with a bifunctional chelator. In certain embodiments, an IL-23R inhibitor of the present disclosure is radiolabeled. The labeled IL-23R inhibitor is then administered to a subject orally or rectally. In certain embodiments, the IL-23R inhibitor is included in drinking water. Following uptake of the IL-23R inhibitor, microPET imaging may be used to visualize inflammation throughout the subject’s bowels and digestive track.

Methods of Treatment and Uses

The present disclosure relates to methods for treating a subject afflicted with a condition or indication associated with IL-23 or IL-23R activity (e.g., activation of the IL-23/IL-23R signaling pathway), wherein the method comprises administering to the subject an IL-23R inhibitor disclosed herein. In one aspect, the present disclosure provides a method for treating a subject afflicted with a condition or indication characterized by aberrant or dysregulated IL-23 or IL-23R activity or signaling, comprising administering to the subject a peptide inhibitor of the present disclosure in an amount sufficient to inhibit (partially or fully) binding of IL-23 to an IL- 23R in the subject. The inhibition of IL-23 binding to IL-23R may occur in particular organs or tissues of the subject, e.g., the stomach, small intestine, large intestine/colon, intestinal mucosa, lamina propria, Peyer’s Patches, mesenteric lymph nodes, or lymphatic ducts.

The present disclosure relates to methods comprising providing a peptide inhibitor described herein to a subject in need thereof. The subject in need thereof may be a subject that has been diagnosed with or has been determined to be at risk of developing a disease or disorder associated with IL-23/IL-23R. The subject may be a mammal. The subject may be, in particular, a human.

The disease or disorder to be treated by treatment with an IL-23R inhibitor of the present disclosure may be an inflammatory disease or disorder, an autoimmune inflammation diseases or disorder, and/or related disorders, including multiple sclerosis, asthma, rheumatoid arthritis, inflammation of the gut, inflammatory bowel diseases (IBDs), juvenile IBD, adolescent IBD, Crohn’s disease, ulcerative colitis, sarcoidosis, Systemic Lupus Erythematosus, ankylosing spondylitis (axial spondyloarthritis), psoriatic arthritis, or psoriasis. In particular, the disease or disorder may be psoriasis (e.g., plaque psoriasis, guttate psoriasis, inverse psoriasis, pustular psoriasis, Palmo-Plantar Pustulosis, psoriasis vulgaris, or erythrodermic psoriasis), atopic dermatitis, acne ectopica, ulcerative colitis, Crohn’s disease, Celiac disease (nontropical Sprue), enteropathy associated with seronegative arthropathies, microscopic colitis, collagenous colitis, eosinophilic gastroenteritis/esophagitis, colitis associated with radio- or chemo-therapy, colitis associated with disorders of innate immunity as in leukocyte adhesion deficiency-1, chronic granulomatous disease, glycogen storage disease type lb, Hermansky-Pudlak syndrome, Chediak-Higashi syndrome, Wiskott-Aldrich Syndrome, pouchitis, pouchitis resulting after proctocolectomy and ileoanal anastomosis, gastrointestinal cancer, pancreatitis, insulindependent diabetes mellitus, mastitis, cholecystitis, cholangitis, primary biliary cirrhosis, viral- associated enteropathy, pericholangitis, chronic bronchitis, chronic sinusitis, asthma, uveitis, or graft versus host disease.

The present disclosure provides a method or use of an IL-23R inhibitor for treating an inflammatory disease or disorder in a subject in need thereof that includes administering to the subject a therapeutically effective amount of an IL-23R inhibitor of the present disclosure, a pharmaceutically acceptable salt thereof, or a composition disclosed herein comprising an IL-23 inhibitor of the present disclosure.

The present disclosure provides a method or use of an IL-23R inhibitor for treating an autoimmune disease or disorder in a subject in need thereof that includes administering to the subject a therapeutically effective amount of an IL-23R inhibitor of the present disclosure, a pharmaceutically acceptable salt thereof, or a composition disclosed herein comprising an IL-23 inhibitor of the present disclosure.

The present disclosure provides a method or use of an IL-23R inhibitor for treating an autoimmune inflammation disease or disorder in a subject in need thereof that includes administering to the subject a therapeutically effective amount of an IL-23R inhibitor of the present disclosure, a pharmaceutically acceptable salt thereof, or a composition disclosed herein comprising an IL-23 inhibitor of the present disclosure.

Suitable inflammatory diseases, autoimmune inflammation diseases, and/or related disorders for treatment with a compound or pharmaceutically acceptable salt thereof, or a composition of the present disclosure, may include, but are not limited to inflammatory bowel disease (IBD), Crohn’s disease (CD), ulcerative colitis (UC), psoriasis (PsO), or psoriatic arthritis (PsA) and the like. The inflammatory disease to be treated may be inflammatory bowel disease (IBD), Crohn’s disease, or ulcerative colitis. The inflammatory disease to be treated may be selected from psoriasis or psoriatic arthritis. The inflammatory disease to be treated may be psoriasis The inflammatory disease to be treated may be psoriatic arthritis. The inflammatory disease to be treated may be IBD. The inflammatory disease to be treated may be Crohn’s disease. The inflammatory disease to be treated may be ulcerative colitis.

Production of IL-23 is enriched in the intestine, where it is believed to play a key role in regulating the balance between tolerance and immunity through T-cell-dependent and T-cell- independent pathways of intestinal inflammation through effects on T-helper 1 (Thl) and Thl7- associated cytokines, as well as restraining regulatory T-cell responses in the gut, favoring inflammation. In addition, polymorphisms in the IL-23 receptor (IL-23R) have been associated with susceptibility to inflammatory bowel diseases (IBDs), further establishing the critical role of the IL-23 pathway in intestinal homeostasis. Peptides and methods for specific targeting of the IL-23R from the luminal side of the gut may provide therapeutic benefit to IBD patients suffering from local inflammation of the intestinal tissue.

Accordingly, the present disclosure also provides a method of treating or preventing inflammatory bowel disease (IBD), Crohn’s disease (CD), or ulcerative colitis (UC), in a subject in need thereof, said method comprising administering to the subject a therapeutically effective amount of a peptide of the present disclosure, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition described herein comprising an IL-23 inhibitor. In some embodiments, the method is for treating or preventing inflammatory bowel disease (IBD). In some embodiments, the method is for treating or preventing Crohn’s disease (CD). In some embodiments, the method is for treating or preventing ulcerative colitis (UC).

Psoriasis, a chronic skin disease affecting about 2%-3% of the general population has been shown to be mediated by the body’s T cell inflammatory response mechanisms. IL-23 is one of several interleukins implicated as a key player in the pathogenesis of psoriasis, purportedly by maintaining chronic autoimmune inflammation via the induction of interleukin- 17, regulation of T memory cells, and activation of macrophages. Expression of IL-23 and IL- 23R has been shown to be increased in tissues of patients with psoriasis, and antibodies that neutralize IL-23 showed IL-23 -dependent inhibition of psoriasis development in animal models of psoriasis. Orally bioavailable peptide inhibitors of IL-23 may provide both a non-steroidal treatment option for patients with mild to moderate psoriasis and treatment for moderate to severe psoriasis that does not require delivery by infusion.

Accordingly, the present disclosure also provides a method of treating or preventing psoriasis (PsO) or psoriatic arthritis (PsA) in a subject in need thereof, said method comprising administering to the subject a therapeutically effective amount of a peptide of the present disclosure, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition described herein comprising an IL-23 inhibitor. In some embodiments, the method is for treating or preventing psoriasis (PsO). In some embodiments, the method is for treating or preventing psoriatic arthritis (PsA).

The present disclosure further relates to a method of selectively inhibiting IL-23 or IL- 23R signaling (or the binding of IL-23 to IL-23R) in a subject (e.g., in a subject in need thereof), comprising administering to the subject a peptide inhibitor of the IL-23R described herein. In some embodiments, the present disclosure includes and provides a method of selectively inhibiting IL-23 or IL-23R signaling (or the binding of IL-23 to IL-23R) in the GI tract of a subject (e.g., a subject in need thereof), comprising providing to the subject a peptide inhibitor of the IL-23R of the present disclosure by oral administration. The exposure of GI tissues (e.g., small intestine or colon) to the administered peptide inhibitor may be at least 10-fold, at least 20- fold, at least 50-fold, or at least 100-fold greater than the exposure (level) in the blood. In particular embodiments, the present disclosure includes a method of selectively inhibiting IL23 or IL23R signaling (or the binding of IL23 to IL23R) in the GI tract of a subject (e.g., a subject in need thereof), comprising providing to the subject a peptide inhibitor, wherein the peptide inhibitor does not block the interaction between IL-6 and IL-6R or antagonize the IL- 12 signaling pathway. In a further related embodiment, the present disclosure provides a method of inhibiting GI inflammation and/or neutrophil infiltration to the GI, comprising providing to a subject in need thereof a peptide inhibitor of the present disclosure. In some embodiments, methods of the present disclosure comprise providing a peptide inhibitor of the present disclosure (i.e., a first therapeutic agent) to a subject (e.g., a subject in need thereof) in combination with a second therapeutic agent. In certain embodiments, the second therapeutic agent is provided to the subject before and/or simultaneously with and/or after the peptide inhibitor is administered to the subject. In particular embodiments, the second therapeutic agent is an anti-inflammatory agent. In certain embodiments, the second therapeutic agent is a nonsteroidal anti-inflammatory drug, steroid, or immune modulating agent. In certain embodiments, the method comprises administering to the subject a third therapeutic agent. In certain embodiments, the second therapeutic agent is an antibody that binds IL-23 or IL-23R.

The present disclosure also relates to methods of inhibiting IL-23 binding to an IL-23R on a cell, comprising contacting the IL-23R with a peptide inhibitor of the receptor disclosed herein. The cell may be a mammalian cell. The method may be performed in vitro or in vivo. Inhibition of binding may be determined by a variety of routine experimental methods and assays known in the art. The present disclosure relates to methods of inhibiting IL-23 signaling by a cell, comprising contacting the IL-23R with a peptide inhibitor described herein. In certain embodiments, the cell is a mammalian cell. In particular embodiments, the method is performed in vitro or in vivo. In particular embodiments, the inhibition of IL-23 signaling may be determined by measuring changes in phospho-STAT3 levels in the cell.

Examples

The following examples are not intended to limit the scope of the present disclosure, but rather to provide guidance to the skilled artisan to prepare and use the peptides, compositions, and methods of the present disclosure. While particular aspects of the present disclosure are described, the skilled artisan will appreciate that various changes and modifications can be made without departing from the spirit and scope of the disclosure.

Some abbreviations useful in describing the disclosure are defined below in the following tables. Herein and throughout the application, the following abbreviations may be used.

Table 3. N-Terminal Modification Abbreviations

Table 4. C-Terminal Modification Abbreviations

The amino acid structures provided in Table 5, below, are presented without stereochemical indicators at the alpha carbon; however, it is to be understood that these amino acids occur as either the L-amino acid or the D-amino acid. For example, “Dap” may be present in the peptides of the instant disclosure as the L-stereoisomer: or as the D-stereoisomer (e.g., when referred to as “dap,” “dDap,” or “D-Dap”):

Table 5. Monomer Abbreviations

Example 1: General Procedure for Solid-Phase Synthesis of Peptides Peptide were chemically synthesized using optimized 9-fluorenylmethoxy carbonyl (Fmoc) solid phase peptide synthesis protocols. For C-terminal amides, Rink-amide MB HA resin was used. The side chain protecting groups were as follows: Asp: OA11; Glu: OA11; Thr: O-tButyl; Asn, Pen: Trityl; AEF: Boc. For coupling, a two to five-fold excess of a solution containing Fmoc amino acid, HATU and DIEA (1 : 0.95:2) in DMF was added to swelled resin for 1 to 48 hours. Double coupling is employed when coupling 2Nal. Fmoc protecting group removal was achieved by treatment with a DMF, piperidine (4: 1) solution for 30 min. The cycles are repeated until the full-length peptide is obtained. OA11 protecting group on Glu removal was achieved by treatment with Pd(PPhs)4 (0. leq), PhSiH? (10eq) and DCM solution for 15min*3 times. For amide cyclization, a solution containing DIC (3.0 eq) and HOBT (3.0 eq) in DMF was added to swelled resin for 16 h*3 times. The cycles are repeated until the full-length peptide is obtained. For RCM cyclization, a solution containing Grubbs 1st (0.5 eq) in DCM was added to swelled resin microwave condition at 40°C for 2h*2 times.

Certain materials and reagents are listed below.

General procedure for cleavage of peptides off resin

Side chain deprotection and cleavage of the peptides was achieved by stirring the dry resin in a solution of trifluoroacetic acid, water, DTT and tri-isopropylsilane (90:2.5:5:2.5) for 3 hours. The mixture was then filtered and cold methyl tert-butyl ether (MTBE) was added to the combined filtrate to precipitate the peptide. The resulting mixture was centrifuged (3000 rpm, 3 min) and decanted. The pellet was washed with MTBE and centrifuged. The pellet was lyophilized to provide the linear peptide.

Procedure for ring closing metathesis (RCM)-cyclization

For RCM cyclization, the peptide was dissolved in anhydrous DCM and stirred before addition of the Grubbs 1^st generation catalyst (0.5 eq). The mixture was allowed to react at 40 °C for 16hrs. When LCMS showed the reaction was complete, the reaction mixture was concentrated under reduced pressure to remove solvent.

General procedure for cyclization

To effect cyclization of thiol-containing residues, iodine solution in MeOH (0.1M) was added to a solution of the linear peptide (20% MeCN/H20 (lmmol/L)) drop-wise until a yellow color persisted. After about 2h, analysis by LCMS showed that the linear peptide was no longer present. The excess iodine was quenched by the addition of IM Na2S20s in water (turned colorless instantly).

General procedure for purification of peptides

Purification of the peptides was achieved using reverse-phase high performance liquid chromatography (RP-HPLC). Purification of the cyclized peptides was achieved using preparative RP-HPLC with a C18 column with a flow rate of 20-250 mL/min. Separation was achieved using gradients of buffer B in A (Buffer A: 0.075% TFA in water; Buffer B: ACN).

(Note 1). Analysis was performed using a C18 column with a flow rate of 1 mL/min (Note 2).

Note 1 : Preparative HPLC Methods

Prep. HPLC Method A: Description: Mobile Phase: 0.075% TFA in water (solvent A) and acetonitrile (solvent B) Column: Welch Ultimate® XB-C18, 250*50 mm, 10 um, 120A + Welch Xtimate®C18, 250*50 mm, 10 um, 120A; Flow Rate: 80 mL/min; Wavelength: UV 220nm&254nm; Oven Tern. Room temperature

Prep. HPLC Method B: Description: Mobile Phase: 0.075% TFA in water (solvent A) and acetonitrile (solvent B) Column: YMC-Actus Triart Cl 8, 250*30 mm, 5 um, 120A column; Flow Rate: 20 mL/min; Wavelength: UV 220nm&254nm; Oven Tern. Room temperature

Note 2: Analytical HPLC Method:

Mobile Phase: 0.1% TFA in water (solvent A) and 0.1%TFA in acetonitrile (solvent B), using the elution gradient 10%-80% (solvent B) over 0.9 minutes and using the elution gradient 80%-90% for 0.6 minutes at a flow rate of 1.0 ml/min; Column: Xbridge C18,3.5um,2.1 *30mm; Wavelength: UV 220nm&254nm; Column temperature: 30°C; MS ionization: ESI

Example 2: Synthesis of SEQ ID NO: 3

The peptide was synthesized using standard Fmoc chemistry. 1) DMF and MBHA Resin (0.30 mmol, 0.90 g, sub: 0.33 mmol/g) were combined in a vessel, and the resin was allowed to swell for two hours.

2) A solution of 20% piperidine/DMF was added and the suspension was mixed for 30 min.

3) The resin was then drained and washed with DMF for 30 seconds *5 times.

4) Fmoc-amino acid solution was then added and mixed with the resin for 30 seconds before addition of a solution of HATU and DIEA in DMF. The reaction was allowed to proceed under nitrogen for 1-4 hours.

5) A solution of 20% piperidine/DMF was added and the suspension was mixed for 30 min.

6) Steps 2 to 5 were repeated for subsequent amino acid couplings. The coupling reactions were monitored by ninhydrin or tetrachlor color test, and upon completion, the resin was washed with DMF 5 times. Once peptide synthesis was complete, the resin was washed with MeOH 3 times and dried by vacuum.

Monitoring method:

1. Ninhydrin test: A: 5% ninhydrin ZEtOH; B: 80% phenol ZEtOH; C: pyridine

2. Tetrachlor color test: A: 2% tetrachlor/ DMF; B: 2% aldehyde/ DMF 110°C for 3min

Detail synthetic method for ring closing metathesis (RCM) cyclization:

For RCM cyclization: the resin and Grubbs 1^st catalyst (0.5 eq) in anhydrous DCM were added into a microwave tube. The mixture was then heated at 40 °C for 2 hr*2 times under microwave conditions, after which LCMS showed the desired product. The resin was then washed with DMF 5 times and with MeOH 3 times before drying under vacuum.

Peptide Cleavage:

1) To the flask containing the side chain protected peptide at room temperature was added 30 mL cleavage buffer (5.0% DTT /2.5% H2O /2.5% TIS /90%TFA), and mixture was stirred for 3 hrs.

2) The mixture was filtered and washed with 5 mL TFA. The combined filtrate was triturated with cold methyl tertbutyl ether (MTBE). The mixture was centrifuged (3000 rpm, 3 min) and decanted. The pellet was washed with MTBE and centrifuged.

3) The residue was lyophilized to give intermediate 1 (520 mg, 90.3% yield, crude).

(Intermediate 1)

Peptide Cyclization and Purification: Crude peptide intermediate 1 (520 mg, 0.271 mmol) was dissolved in 20% MeCN /H2O

(300 mL). To a stirred solution of the peptide was added the iodine solution in MeOH (0. IM, 3.5 mL) drop-wise until the color of the solution remains yellow. After ~2h LCMS showed the reaction was complete. Excess iodine was quenched by the addition of IM Na2S20s in water (15 uL) (turned colorless instantly). Then was added 10-20 mL of MeCN to decrease turbidity. The solution was purified the solution by Prep-HPLC (A: 0.075% TFA in H2O, B: ACN) (Note 1 : Method A) to give the peptide of SEQ ID NO: 3 (38.4 mg, 96.4% purity, 6.37% yield for this step; over all yield: 5.75%) obtained as white solid. Analysis was performed using a C18 column with a flow rate of 1 mL/min (Note 2). LCMS Summary: Method: 10-80-2min-l.5_P2.amx, retention time: 1.518 min, calculated MW: 1917.25, observed MW: 959.3 [(M+2H)/2],

Example 3: Synthesis of SEQ ID NO: 2

The peptide was synthesized using standard Fmoc chemistry.

1) DMF and MBHA Resin (0.30 mmol, 0.90 g, sub: 0.33 mmol/g) were combined in a vessel, and the resin was allowed to swell for two hours.

3) The resin was then drained and washed with DMF for 30 seconds *5 times.

Monitoring method:

1. Ninhydrin test: A: 5% ninhydrin ZEtOH; B: 80% phenol ZEtOH; C: pyridine

2. Tetrachlor color test: A: 2% tetrachlor/ DMF; B: 2% aldehyde/ DMF 110°C for 3min Detail synthetic method for ring closing metathesis (RCM) cyclization:

Peptide Cleavage:

3) The residue was lyophilized to give intermediate 2 (500 mg, 90.6% yield, crude).

(Intermediate 2)

Peptide Cyclization and Purification:

Crude peptide intermediate 2 (500 mg, 0.272 mmol) was dissolved in 20% MeCN /H2O (300 mL). To a stirred solution of the peptide was added the iodine solution in MeOH (0. IM, 3.5 mL) drop-wise until the color of the solution remains yellow. After ~2h LCMS showed the reaction was complete. Excess iodine was quenched by the addition of IM Na2S20s in water (15 uL) (turned colorless instantly). Then was added 10-20 mL of MeCN to decrease turbidity. The solution was purified by Prep-HPLC (A: 0.075% TFA in H2O, B: ACN) (Note 1 : Method A) to give the peptide of SEQ ID NO: 2 (38.3 mg, 98.0% purity, 6.69% yield for this step; over all yield: 6.06%) obtained as white solid. Analysis was performed using a C18 column with a flow rate of 1 mL/min (Note 2).

LCMS Summary: Method: 10-80-2min-l.5_P2.amx, retention time: 1.497 min, calculated MW: 1837.13, observed MW: 919.2[(M+2H)/2],

Example 4: Synthesis of SEQ ID NO: 5

The peptide was synthesized using standard Fmoc chemistry.

1) DMF and MBHA Resin (0.30 mmol, 0.96 g, sub: 0.31 mmol/g) were combined in a vessel, and the resin was allowed to swell for two hours.

3) The resin was then drained and washed with DMF for 30 seconds *5 times.

Monitoring method:

1. Ninhydrin test: A: 5% ninhydrin ZEtOH; B: 80% phenol ZEtOH; C: pyridine

Detail synthetic method for amide cyclization:

De-OAll on Glu: The resin was washed with 50 mL DMF (3x0.1 min) and DCM (3x0.1 min) before addition of PhSiEEQO eq) and Pd(PPhs)4 (0.1 eq) in DCM (10 mL). The mixture was reacted for 15 min and washed with DCM (50 mL) and DMF (50 mL) alternately 5 times until the solution turned colorless.

Amide cyclization on resin: After de-protection, the resin was washed with 50 mL of DMF (5x0.1 min) followed by addition of DIC (3 eq) and HOBT(3.0 eq) in DMF (50 mL). The coupling reaction was mixed for 16h. Ninhydrin color reaction showed negative test. After completing the coupling reaction, the resin was washed with 50 mL DMF (3x0.1 min).

Peptide Cleavage:

1) To the flask containing the side chain protected peptide at room temperature was added 30 mL cleavage buffer (5.0% DTT /2.5% FEO /2.5% TIS /90%TFA), and mixture was stirred for 3 hrs.

3) The residue was lyophilized to give intermediate 3 (550 mg, 92.5% yield, crude).

Peptide Cyclization and Purification: Crude peptide intermediate 3 (550 mg, 0.278 mmol) was dissolved in 20% MeCN /H2O

(300 mL). To a stirred solution of the peptide was added the iodine solution in MeOH (0. IM, 2.5 mL) drop-wise until the color of the solution remains yellow. After ~2h LCMS showed the reaction was complete. Excess iodine was quenched by the addition of IM Na2S20s in water (15 uL) (turned colorless instantly). The was added 10-20 mL of MeCN to decrease turbidity. The solution was purified by Prep-HPLC (A: 0.075% TFA in H2O, B: ACN) (Note 1 : Method B) to give the peptide of SEQ ID NO: 5 (19.3 mg, 98.5% purity, 3.10% yield for this step; over all yield: 2.87%) obtained as white solid. Analysis was performed using a C18 column with a flow rate of 1 mL/min (Note 2). LCMS Summary: Method: 10-80-3min-l.5_P2.amx, retention time: 1.384 min, calculated MW: 1979.33, observed MW: 990.2[(M+2H)/2],

Example 5: Synthesis of SEQ ID NO: 4

The peptide was synthesized using standard Fmoc chemistry.

1) DMF and MBHA Resin (0.5 mmol, 1.5 g, sub: 0.33 mmol/g) were combined in a vessel, and the resin was allowed to swell for two hours. 2) A solution of 20% piperidine/DMF was added and the suspension was mixed for 30 min.

3) The resin was then drained and washed with DMF for 30 seconds *5 times.

4) Fmoc-amino acid solution was then added and mixed with the resin for 30 seconds before addition of a solution of HATU and DIEA in DMF. The reaction was allowed to proceed under nitrogen for 1-4 hours. 5) A solution of 20% piperidine/DMF was added and the suspension was mixed for 30 min.

Monitoring method: 1. Ninhydrin test: A: 5% ninhydrin ZEtOH; B: 80% phenol ZEtOH; C: pyridine

Peptide Cleavage: 1) To the flask containing the side chain protected peptide at room temperature was added 30 mL cleavage buffer (5.0% DTT /2.5% FEO /2.5% TIS /90%TFA), and mixture was stirred for 3 hrs.

3) The residue was lyophilized to give intermediate 4a (0.9 g, 74.6% yield, crude).

(Intermediate 4a) Peptide Cyclization and Purification: Crude peptide intermediate 4a (900 mg, 0.373 mmol) was dissolved in 20% MeCN

/H2O (500 mL). To a stirred solution of the peptide was added the iodine solution in MeOH (0.1M, 7.5 mL) drop-wise until the color of the solution remains yellow. After ~2h LCMS showed the reaction was complete. Excess iodine was quenched by the addition of IM Na2S20s in water (20 uL) (turned colorless instantly). Then was added 10-20 mL of MeCN to decrease turbidity. The solution was purified by Prep-HPLC (A: 0.075% TFA in H2O, B: ACN) (Note 1 : Method B) to give intermediate 4b (190 mg, 21.1% yield) obtained as white solid. Analysis was performed using a C18 column with a flow rate of 1 mL/min (Note 2).

(Intermediate 4b)

Peptide intermediate 4b (190 mg, 0.079 mmol) was dissolved in anhydrous DCM. To a stirred solution of the peptide was added the Grubbs 1^st cata^lyst (33 mg, 0.5 eq) and the mixture was allowed to react at 40°C for 16 hrs, at which time, LCMS showed the reaction was complete. The reaction mixture was concentrated under reduced pressure to remove solvent and purified by Prep-HPLC (A: 0.075% TFA in H2O, B: ACN) (Note 1 : Method B) to give the peptide of SEQ ID NO: 4 (10.5 mg, 94.8% purity, 4.64% yield for this step; over all yield:

0.73%) obtained as white solid. Analysis was performed using a C18 column with a flow rate of 1 mL/min (Note 2).

LCMS Summary: Method: 10-80-2min-l.5_P2.amx, retention time: 1.565 min, calculated MW: 2380.95, observed MW: 1190.1[(M+2H)/2],

Example 6: IL23R Reporter Assay

Compounds were serially diluted in 100% (v/v) DMSO) and plated using an Echo acoustic dispenser (Labcyte) into 1536-well non-treated black assay plates (Coming # 9146). 3 pL of HEK293 cells containing IL-23R, IL-12RP1 and a firefly luciferase reporter gene driven by a STAT -inducible promoter (Promega) were added to the plates (4000 cells/well), followed by 3 pL of 10 ng/mL IL-23 (equivalent to EC90 concentration). After 5h at 37°C, 5% CO2, 95% relative humidity, cells were placed at 20°C and treated with BioGio reagent (Promega) according to the Manufacturer’s instructions. Luminescence was measured on a Pherastar FSX (BMG LabTech). Data were normalized to IL-23 treatment (0% inhibition) and 30 pM of control inhibitor (100% inhibition), and IC50 values were determined using a 4-parameter Hill equation. Data for example compounds are shown below.

A: IC₅o < O.Ol pM;

B: 0.01 pM < IC₅o < O.5 pM;

C: 0.5 pM < IC₅o

ND: Not determined

Table 6. IL-23 Binding Data

Example 7: PBMC pSTAT3 Assay Cryopreserved peripheral blood mononuclear cells (PBMCs) from healthy donors were thawed and washed twice in ImmunoCult-XF T cell expansion medium (XF-TCEM) supplemented with CTL anti-aggregate wash. The cells were counted, resuspended at 2-6xl0⁵ cells per mL XF-TCEM supplemented with penicillin/ streptomycin and 100 ng/mL IL-ip (BioLegend, 579404), and cultured in tissue culture flasks coated with anti-CD3 (eBioscience, 16-0037-85 or BD Pharmingen, 555329) at 37oC in 5% CO2. On day 4 of culture, PBMCs were collected, washed twice in RPMI-1640 supplemented with 0.1% BSA (RPMI-BSA), and incubated in RPMI-BSA in upright tissue culture flasks for ~4 hours at 37oC in 5% CO2. Following this ‘starvation,’ a total of 6x104 cells in 30 pL RPMI-BSA was transferred into each well of a 384-well plate pre-spotted with peptide or DMSO. The cells were incubated for 30 minutes prior to the addition of IL-23 at a final concentration of 5 ng/mL. The cells were stimulated with cytokine for 30 minutes at 37oC in 5% CO2, transferred onto ice for 10 minutes, and lysed. Cell lysates were stored at -80°C until phosphorylated STAT3 was measured using the phospho-STAT panel kit (Meso Scale Discovery, K15202D). Results are provided below.

A: IC₅o < 1 nM;

B: 1 nM < IC50 < 10 nM;

C: 10 nM < IC₅₀;

ND: Not determined

Table 7. PBMC pSTAT3 Assay Results

While the foregoing specification teaches the principles of the present invention, with examples provided for the purpose of illustration, it will be understood that the practice of the invention encompasses all of the usual variations, adaptations and/or modifications as come within the scope of the following claims and their equivalents.

Claims

CLAIMS What is claimed is:

1. A peptide of Formula (I’), comprising the amino acid sequence:

Ri is MeCO, 8Aoc, 7Ahp, 6Ahx, 5Ava, or cPEG3aCO;

X3 is hK, a ring-forming amino acid, or absent;

X4 is any amino acid;

X5 is N, N(NMe2), Q, Q(NMe2), or a ring-forming amino acid;

Xe is any amino acid;

X₇ is 7MeW or W;

X₈ is K(Ac), K(NMeAc), Q, or a ring-forming amino acid;

X9 is any amino acid;

X10 is AEF, APEG3F, F(4TzlAme2), TMAPF, or a ring-forming amino acid;

X12 is THP, or a ring-forming amino acid;

X13 is E or a ring-forming amino acid;

X15 is 3Pya, bAla, or a ring-forming amino acid;

Xi6 is Sar, a ring-forming amino acid, or absent;

R₂ is CONH2 or CONMe₂; wherein:

(a) a first ring-forming amino acid is linked to a second ring-forming amino acid to form a first ring comprising 4-11 or 14 amino acids; and a third ring-forming amino acid is linked to a fourth ring forming amino acid to form a second ring comprising 4-11 or 14 amino acids; or

2. The peptide of claim 1, wherein the peptide comprises the amino acid sequence of Formula (I):

Ri-X3-X4-X₅-T-X7-X8-X₉-Xio-2Nal-Xi2-Xi3-N-Xi₅-Xi6-R2 (I), or a pharmaceutically acceptable salt thereof, wherein: Ri is MeCO, 8Aoc, 7Ahp, or cPEG3aCO;

X₃ is R7H, S7H, hK, or absent;

X₅ is N, N(NMe2), Q, or Q(NMe2);

X₇ is 7MeW or W;

X₈ is K(Ac), R5H, S5H, K(NMeAc), or Q;

X₉ is aMeC, aG, C, D, E, hE, Pen, or Dap(N3);

X10 is AEF, APEG3F, F(4TzlAme2), or TMAPF;

Xn is R5, S5, B5, or THP;

X13 is E, R5H, or S5H;

X15 is 3Pya or bAla;

Xi6 is R5H, S5H, Sar, or absent;

R₂ is CONH2 or CONMe₂; wherein when X3 is R7H or S7H, then X5 is N(Me2) or Q(NMe2), X₈ is K(NmeAc), or X10 is APEG3F; wherein the peptide is cyclized via:

(a) a first linkage between the residues at X4 and X₉; and

(b) a second linkage selected from the group consisting of a linkage between 8Aoc at Ri and E at X13, a linkage between 7Ahp at Ri and E at X13, a linkage between R7H or S7H at X3 and R5H or S5H at X13, a linkage between hk at X3 and E at X13, a linkage between R5H or S5H at X₈ and R5 or S5 at X12, a linkage between F(4TzlAme2) at X10 and E at X13, a linkage between AEF at X10 and E at X13; a linkage between R5 or S5 at X12 and R5H or S5H at Xie; and two linkages between B5 at X12 and R5H or S5H at X₈ and between B5 at X12 and R5H or S5H at Xie.

3. The peptide of claim 1 or claim 2, or a pharmaceutically acceptable salt thereof, wherein when X3 is hk, then X15 is bAla;

4. The peptide of claim 1 or claim 2, or a pharmaceutically acceptable salt thereof, having an amino acid sequence of Formula (I-A): Ri-X3-X₄-X₅-T-X7-X8-X₉-Xio-2Nal-Xi2-Xi3-N-3Pya-Xi6-R2 (I-A) wherein:

Ri is MeCO, 8Aoc, or cPEG3aCO;

Xs is N or N(NMe2); and

Xi6 is S5H or Sar.

5. The peptide of claim 1 or claim 2, or a pharmaceutically acceptable salt thereof, having an amino acid sequence of Formula (I-B):

Ri-X3-Pen-X₅-T-7MeW-X₈-Pen-Xio-2Nal-Xi2-Xi3-N-3Pya-Xi6-R2 (LB) wherein:

Ri is MeCO, 8Aoc, or cPEG3aCO;

X₅ is N or N(NMe₂);

X₈ is K(Ac), S5H, or K(NMeAc); and

Xi6 is S5H or Sar.

6. The peptide of claim 1 or claim 2, or a pharmaceutically acceptable salt thereof, having an amino acid sequence of Formula (I-C):

Ri-X3-X₄-X₅-T-X7-X₈-X₉-Xio-2Nal-THP-Xi3-N-3Pya-Xi6-R2 (I-C) wherein:

X₈ is K(Ac), K(NMeAc), or Q; and

Xi6 is Sar or absent.

7. The peptide of claim 1 or claim 2, or a pharmaceutically acceptable salt thereof, having an amino acid sequence of Formula (I-D):

Ri-X3-X₄-X₅-T-X₇-X₈-X₉-Xio-2Nal-Xi2-E-N-3Pya-Xi6-CONH2 (I-D) wherein:

Ri is MeCO or 8Aoc;

X3 is absent;

X5 is N or Q;

X₈ is K(Ac), S5H, or Q;

X10 is AEF or F(4TzlAme2); and

Xi6 is S5H or Sar.

8. The peptide of claim 1 or claim 2, or a pharmaceutically acceptable salt thereof, having a sequence according to any one of Formulas I-E to I-K: Ri-X3-Pen-X₅-T-7MeW-X₈-Pen-Xio-2Nal-Xi2-E-N-3Pya-Xi6-R2 (I-E) Ri-X3-X₄-X₅-T-X₇-X₈-X9-AEF-2Nal-THP-Xi3-N-3Pya-R2 (I-F) Ri-X3-X₄-X₅-T-X7-X₈-X₉-AEF-2Nal-Xi2-E-N-3Pya-CONH2 (I-J) MeCO-X3-X₄-X₅-T-X₇-X₈-X₉-AEF-2Nal-THP-E-N-3Pya-R2 (I-K).

9. The peptide of any one of claims 1-7, or a pharmaceutically acceptable salt thereof, wherein:

X₄ is Pen and X₉ is Pen; or

X₄ is Abu and X₉ is C.

10. The peptide of claim 2, or a pharmaceutically acceptable salt thereof, wherein the second linkage is an amide linkage between 8Aoc at Ri and E at X13.

11. The peptide of claim 2, or a pharmaceutically acceptable salt thereof, wherein the second linkage is an amino linkage between AEF at Xw and E at X13.

12. The peptide of claim 2, or a pharmaceutically acceptable salt thereof, wherein the second linkage is an aliphatic linkage between R7H or S7H at X3 and R5H or S5H at X13.

13. The peptide of claim 2, or a pharmaceutically acceptable salt thereof, wherein the second linkage is an amide linkage between hK at X3 and E at X13.

14. The peptide of claim 2, or a pharmaceutically acceptable salt thereof, wherein the second linkage is an aliphatic linkage between R5H or S5H at X₈ and R5 or S5 at X12.

15. The peptide of claim 2, or a pharmaceutically acceptable salt thereof, wherein the second linkage is an ester linkage between F(4TzlAme2) at Xw and E at X13.

16. The peptide of claim 2, or a pharmaceutically acceptable salt thereof, wherein the second linkage is an aliphatic linkage between R5 or S5 at X12 and R5H or S5H at Xi6.

17. The peptide of claim 2, or a pharmaceutically acceptable salt thereof, wherein the second linkage is two aliphatic linkages: between B5 at X12 and R5H or S5H at X₈; and between B5 at X12 and R5H or S5H at Xi6.

18. The peptide of any of claims 1-9 or 12-17, or a pharmaceutically acceptable salt thereof, wherein Ri is MeCO.

19. The peptide of any of claims 1-18, or a pharmaceutically acceptable salt thereof, wherein R₂ is C0NH₂.

20. The peptide of claim 1, wherein the first ring comprises 4-9 or 11 amino acids.

21. The peptide of claim 1, wherein first ring is formed between X4 and X9, X4 and X13, X5 and X10, X3 and X13, or Xe and X9.

22. The peptide of claim 21, wherein the first ring is formed between X4 and X9, X4 and X13, or Xe and X9.

23. The peptide of claim 22, wherein the first ring is formed between X4 and X9 via a linker having one or more groups selected from the group consisting of a disulfide, thioether, amide, olefin, ether, alkylene, and triazole.

24. The peptide of claim 22, wherein the first ring is formed between X4 and X13 via a linker having one or more groups selected from the group consisting of a disulfide, thioether, amide, olefin, ether, alkylene, and triazole.

25. The peptide of claim 22, wherein the first ring is formed between Xe and X9 via a linker having one or more groups selected from the group consisting of a disulfide, thioether, amide, olefin, ether, alkylene, and triazole.

26. The peptide of any one of claims 1 or 20-25, wherein the second ring comprises 4, 6, 10, or 11 amino acids.

27. The peptide of any one of claims 1 or 20-26, wherein the second ring is formed between X5 and X10, X3 and X13, or between X13 and the N-terminus of the peptide.

28. The peptide of claim 23, wherein: the second ring is formed between X3 and X13 via a linker having one or more groups selected from the group consisting of a disulfide, thioether, amide, olefin, ether, alkylene, and tri azole; the second ring is formed between X5 and X10 via a linker having one or more groups selected from the group consisting of a disulfide, thioether, amide, olefin, ether, alkylene, and tri azole; the second ring is formed between X10 and X13 via a linker having one or more groups selected from the group consisting of a disulfide, thioether, amide, olefin, ether, alkylene, and triazole; or the second ring is formed between X13 and the N-terminus of the peptide via a linker having one or more groups selected from the group consisting of a disulfide, thioether, amide, olefin, ether, alkylene, and triazole.

29. The peptide of claim 24, wherein: the second ring is formed between X5 and X10 via a linker having one or more groups selected from the group consisting of a disulfide, thioether, amide, olefin, ether, alkylene, and triazole; or the second ring is formed between Xe and X9 via a linker having one or more groups selected from the group consisting of a disulfide, thioether, amide, olefin, ether, alkylene, and tri azole.

30. The peptide of claim 25, wherein: the second ring is formed between X3 and X13 via a linker having one or more groups selected from the group consisting of a disulfide, thioether, amide, olefin, ether, alkylene, and tri azole; the second ring is formed between X4 and X13 via a linker having one or more groups selected from the group consisting of a disulfide, thioether, amide, olefin, ether, alkylene, and tri azole; the second ring is formed between X5 and X10 via a linker having one or more groups selected from the group consisting of a disulfide, thioether, amide, olefin, ether, alkylene, and tri azole; the second ring is formed between X10 and X13 via a linker having one or more groups selected from the group consisting of a disulfide, thioether, amide, olefin, ether, alkylene, and triazole; or the second ring is formed between X13 and the N-terminus of the peptide via a linker having one or more groups selected from the group consisting of a disulfide, thioether, amide, olefin, ether, alkylene, and triazole.

31. A peptide having an amino acid sequence of any one of SEQ ID NOS: 1-6 or 8, or a pharmaceutically acceptable salt thereof.

32. A pharmaceutical composition comprising a peptide of any one of claims 1-19, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

33. A method for treating a disease or disorder associated with Interleukin 23 (IL- 23)/Interleukin 23 Receptor (IL-23R), comprising administering to a subject in need thereof a therapeutically effective amount of a peptide of any one of claims 1-19 or a pharmaceutical composition of claim 20.

34. The method of claim 21, wherein the disease or disorder is selected from multiple sclerosis, asthma, rheumatoid arthritis, inflammation of the gut, inflammatory bowel diseases (IBDs), juvenile IBD, adolescent IBD, Crohn’s disease, ulcerative colitis, Celiac disease (nontropical Sprue), microscopic colitis, collagenous colitis, eosinophilic gastroenteritis/esophagitis, colitis associated with radio- or chemo-therapy, colitis associated with disorders of innate immunity as in leukocyte adhesion deficiency-1, sarcoidosis, Systemic Lupus Erythematosus, ankylosing spondylitis (axial spondyloarthritis), psoriatic arthritis, psoriasis (e.g., plaque psoriasis, guttate psoriasis, inverse psoriasis, pustular psoriasis, Palmo- Plantar Pustulosis, psoriasis vulgaris, or erythrodermic psoriasis), atopic dermatitis, acne ectopica, enteropathy associated with seronegative arthropathies, chronic granulomatous disease, glycogen storage disease type lb, Hermansky-Pudlak syndrome, Chediak-Higashi syndrome, Wiskott-Aldrich Syndrome, pouchitis, pouchitis resulting after proctocolectomy and ileoanal anastomosis, gastrointestinal cancer, pancreatitis, insulin-dependent diabetes mellitus, mastitis, cholecystitis, cholangitis, primary biliary cirrhosis, viral-associated enteropathy, pericholangitis, chronic bronchitis, chronic sinusitis, asthma, uveitis, or graft versus host disease.

35. The method of claim 21, wherein the disease or disorder is selected from ulcerative colitis (UC), Crohn’s disease (CD), psoriasis (PsO), or psoriatic arthritis (PsA).