Novel peptides as selective IL-23 receptor inhibitors

Abstract

The present disclosure relates to IL-23 receptor inhibitor compounds of formula (I), pharmaceutical compositions thereof and medical uses thereof, for example in the treatment of inflammatory bowel diseases such as Crohn's disease.

Description

Technical Field

[0001] This disclosure relates to peptide IL-23 receptor inhibitors and their medical uses, such as in the treatment of inflammatory bowel diseases like Crohn's disease. More specifically, this disclosure relates to cyclic peptide compounds that can be used as IL-23 receptor inhibitors. Background Technology

[0002] Mounting evidence suggests that the interleukin-23 (IL-23) cytokine plays a crucial role in the pathogenesis of autoimmune inflammation and related diseases and disorders such as asthma, rheumatoid arthritis, psoriasis, multiple sclerosis, and inflammatory bowel disease (IBD) (e.g., ulcerative colitis and Crohn's disease). The major role of IL-23R can be elucidated through downstream effector cytokines in acute and chronic IBD mouse models and disease pathogenesis. Th17 cells, Y8 T cells, natural killer (NK) cells, dendritic cells, macrophages, and innate lymphoid cells, abundantly found in the gut, are adaptive and innate immune cells that express IL-23R. IBD patients exhibit increased gene expression and IL-23R levels on the intestinal mucosal surface. Evidence suggests that IL-23 contributes to this role by promoting the development of pathogenic CD4+ T cell populations that produce IL-6, IL-17, and tumor necrosis factor (TNF).

[0003] Increased concentrations of IL-23 in the gut are thought to play a crucial role in regulating the balance between tolerance and immunity through both T-cell-dependent and T-cell-independent pathways related to intestinal inflammation. This regulation influences helper T cell 1 (Th1) and Th17-related cytokines and suppresses regulatory T-cell responses in the gut, thereby promoting inflammation. Furthermore, the association of IL-23R polymorphism with susceptibility to intestinal inflammation further confirms the importance of the IL-23 pathway in intestinal homeostasis.

[0004] Psoriasis is a chronic skin disease with an incidence of approximately 2%-3% in Caucasian populations, and its pathogenesis has been shown to be mediated by the body's T-cell inflammatory response. IL-23, one of several interleukins, is believed to play a key role in the pathogenesis of psoriasis by inducing interleukin-17, activating macrophages, and regulating T memory cells. These interleukins are thought to play a crucial role in maintaining chronic autoimmune inflammation. In animal models of psoriasis, neutralizing antibodies against IL-23 have shown IL-23-dependent inhibition of psoriasis development, and increased expression of IL-23 and IL-23R has been demonstrated in tissues of psoriasis patients.

[0005] IL-23 is part of the IL-12 cytokine family and consists of the p19 subunit, which is specific to IL-23, and the p40 subunit, of IL-12, a cytokine involved in the development of helper T cell 1 (TH1) cells that produce interferon-γ (IFN-γ). Although both IL-23 and IL-12 contain the p40 subunit, they exhibit different phenotypic characteristics. While animals lacking IL-12 are susceptible to inflammatory autoimmune diseases, animals lacking IL-23 are resistant, likely due to a reduced number of CD4+ T cells in the CNS that produce IL-6, IL-17, and TNF. IL-23 binds to IL-23R, a heterodimeric receptor composed of the IL-12Rβ1 and IL-23R subunits. The binding of IL-23 to IL-23R activates the Jak-Stat signaling molecules Jak2, Tyk2, Stat1, Stat3, Stat4, and Stat5, although Stat4 activation is significantly weaker, and a different DNA-binding Stat complex is formed in response to IL-23 compared to IL-12. IL-23R is constitutively associated with Jak2 and associated with Stat3 in a ligand-dependent manner. Unlike IL-12, which primarily acts on naive CD4(+) T cells, IL-23 preferentially acts on memory CD4(+) T cells.

[0006] Due to its biological importance, inhibiting the IL-23 pathway is an attractive option for treating IL-23-related diseases and disorders. Numerous antibodies that bind to IL-23 or IL-23R have been identified, including ustekinumab, a humanized antibody that binds to IL-23 and has been approved for the treatment of psoriasis. Recently, peptide inhibitors that bind to IL-23R and inhibit the binding of IL-23 to IL-23R have been identified (see, for example, U.S. Patent Application Publication No. US 2013 / 0029907). Clinical trials using uteroclosmab and briakinumab (which target a common p40 subunit) and tildrakizumab, guselkumab, brazikumab, and risankizumab (which target a unique p19 subunit of IL-23) in Crohn's disease or psoriasis highlight the potential of IL-23 signaling blockade in the treatment of human inflammatory diseases.

[0007] While these findings are promising, challenges remain in identifying stable and selective agents that preferentially target the IL-23 pathway in the gut for the treatment of intestinal inflammation, such as bowel diseases including Crohn's disease, ulcerative colitis, and related disorders.

[0008] Patients with irritable bowel disease (IBD) suffering from intestinal autoimmune inflammation can benefit from oral to local treatments in which compounds specifically target the IL-23 pathway on the luminal side of the intestine.

[0009] The problem associated with the use of these compounds as orally administered therapeutic agents for the treatment of inflammatory diseases and other indications may be the limited stability of these compounds in the intestinal environment.

[0010] Therefore, when developing novel therapeutic molecules, there is a need for variants with improved drug properties (e.g., increased stability against proteases present in the gut and / or increased chemical or physical stability and / or prolonged in vivo half-life and / or increased in vivo potency / efficacy).

[0011] In addition, there is still a need for anti-inflammatory therapies that avoid or mitigate common systemic side effects (e.g., infection defense) of IL-23R-based therapies, thereby achieving anti-inflammatory effects with improved tolerability.

[0012] US 2013-029907 A1 discloses a linear peptide inhibitor of the IL-23 receptor.

[0013] Protagonist's WO 2016 / 011208 A1, WO 2017 / 011820 A2, WO 2018 / 089693A2, WO 2018 / 022937 A1, WO 2018 / 136646 A1, and WO 2022 / 109328 A1, and Sayago et al. (ACSMed. Chem. Lett. [ACS Medicinal Chemistry Letters] 2018, 9, 912-916) disclosed peptide inhibitors of the IL-23 receptor for oral administration. A common structural feature of the disclosed compounds is a ring formed by six amino acids as ring members.

[0014] WO 2013 / 063468 discloses cyclic peptides that are modified with long-chain hydrocarbon groups to produce amphiphilic molecules that can be used as drug delivery systems, including nucleotide delivery to cells.

[0015] WO 2015 / 179438 A1 also discloses cyclic peptides consisting of 2 to 10-membered rings formed from amino acids as inhibitors of Rac or Rho in cells or tissues, wherein at least two amino acids are arginine.

[0016] Quiniou et al. (Am J Physiol Regul Integr Comp Physiol [American Journal of Physiological Regulation, Integrative and Comparative Physiology], 2014, 307: R1216-R1230) disclosed a non-competitive inhibitor of small peptides made from D-amino acids.

[0017] Kuchar et al. (Proteins, 2013) disclosed an IL-23 receptor inhibitor based on a triple-helix scaffold derived from the albumin-binding domain of streptococcal protein G.

[0018] WO 2023 / 288028 discloses a cyclic peptide inhibitor of the IL-23 receptor consisting of 16 amino acid residues, wherein cyclization is achieved via a disulfide bond between residues at positions 4 and 9.

[0019] WO 2023 / 288019 discloses a cyclic peptide inhibitor of the IL-23 receptor consisting of 16 amino acid residues, wherein cyclization is achieved through a disulfide bond or thioether bond between residues at positions 4 and 9.

[0020] WO 2023 / 288017 discloses a bicyclic peptide inhibitor of the IL-23 receptor, wherein bicyclization is achieved through a first disulfide or thioether bond between the first pair of amino acid residues and a second amide or thioether bond between the second pair of amino acid residues.

[0021] Therefore, there remains a need for improved IL-23 receptor inhibitor compounds for the treatment and / or prevention of autoimmune or inflammatory diseases.

[0022] There is a need for IL-23 receptor inhibitor compounds with improved in vitro and in vivo stability.

[0023] There is a need for IL-23 receptor inhibitor compounds that have improved oral bioavailability and / or stability when administered orally.

[0024] IL-23 receptor inhibitor compounds that selectively bind to IL-23 receptors in the gut are needed.

[0025] Compounds with improved IL-23 inhibitor properties are needed.

[0026] There is a need for IL-23 receptor inhibitor compounds with high binding affinity.

[0027] Compounds that can inhibit the binding of IL-23 to its receptor are needed, among which IC50... 50 Within the nanomolar range, for example, equal to or less than 50 nM, or again equal to or less than 20 nM.

[0028] There is a need for IL-23 receptor inhibitor compounds with improved protease stability.

[0029] This disclosure provides peptide compounds of formula (I) that inhibit IL-23 binding, and some embodiments are suitable for oral administration and / or addressing one or more of the above-mentioned needs.

[0030] Therefore, this disclosure provides a compound of formula (I) with affinity for the IL-23 receptor. Summary of the Invention

[0031] This disclosure relates to a peptide compound of formula (I):

[0032] X1(§1)(#1)-X2(#1)(@2)-X3-X4-X5-X6-X7-X8-X9(§1)(@2)-X10-X11-X12-X13-R1(I),

[0033] in

[0034] X1 is absent or has a butyl portion and

[0035] When X1 is a butyl group, then (§1) represents the alkyl bond between X1 and X9, and (#1) represents the alkyl bond between X1 and X2, and in such a case, (@2) does not exist, or

[0036] When X1 is absent, then (@2) represents the amide bond between X2 and X9, and in such a case, (§1) and (#1) are absent.

[0037] X2 is an amino acid residue selected from the following: Add, Ade, Ahd, Aoa, Atd, Atea, Aud, Dad, Dae, Dec, Dod, Dpa, Kme, Kme(αAc), Lys(αAc), Nmapt, Nmha, and Oca.

[0038] X3 is an amino acid residue selected from the following: Nak, Trp, Wdm, Wim, Wme, Wcl, and Wfl;

[0039] X4 is an amino acid residue selected from the following: Aib, Gln, Glu, Ile, Iva, Leu, Lys, Lys(Ac), Mle, Mly, Mva, Phe, Thr, Trp, and Val;

[0040] X5 is an amino acid residue selected from the following: Aib, Mkdm, Mle, and Thp;

[0041] X6 is an amino acid residue selected from the following: Trp, Tyr, Yae, Yde, and Yme;

[0042] X7 is an amino acid residue selected from the following: Nak, Nal, and Trp;

[0043] X8 is an amino acid residue selected from the following: Aib, Cba, Cit, Gln, Leu, Lys, Lys(Ac), Mle, Mly, D-Mkdm, Mkdm, Thp, D-Trp, and D-Tza;

[0044] X9 is an amino acid residue selected from the following: Asp, Glu, Kme, Lys, and Orn;

[0045] X10 is an amino acid residue selected from the following: Ala, Asn, Gly, and Ser;

[0046] X11 is an amino acid residue selected from the following: Ala, Asn, Bal, Gly, Hol, Hph, His, Ile, Iva, Leu, Mhis, Pal, Pyal, PyEA, Val, and their corresponding D-forms;

[0047] X12 is absent or consists of amino acid residues selected from the following: Bal, Dnmy, Ile, Lys, Mep, Mys, and Sar;

[0048] X13 is absent or is a Lys residue; and...

[0049] R1 either does not exist or is selected from -NH2, -OH and -N(C2H5)2;

[0050] Or its salts or solvates.

[0051] The peptide compound of formula (I) inhibits the binding of interleukin-23 (IL-23) to its receptor (IL-23R).

[0052] In some embodiments, the salts or solvates of the peptide compounds disclosed herein are pharmaceutically acceptable salts or solvates.

[0053] As shown in the Examples section, it has been observed that, in some embodiments, peptide compounds of general formula (I) exhibit good solubility in water at a variety of gastrointestinal pH levels (pH 1.2 to 7.4), good stability at pH levels down to 1.2 with a purity loss of less than about 20%, or even less than 15% purity loss in a pH range of 1.1 to 7.4. This capability enables the peptide compounds of this disclosure to be administered orally, wherein in the digestive tract, the pH ranges from about 7.4 in the mouth to about 1.2 in the stomach. Furthermore, some compounds exhibit good stability in simulated gastric and intestinal fluids, with at least 50% of the peptide compounds remaining in the simulated intestinal fluid after 60 min at 37°C. In some embodiments, the peptide compounds of this disclosure have the ability to inhibit the binding of IL-23 to its IL-23 receptor, wherein IC 50 Within the nanomolar range, at least below about 20 nM, or at least below about 10 nM. In some embodiments, the peptide compounds disclosed herein have the ability to inhibit IL-23-induced STAT3 phosphorylation, wherein IC 50 Within the nanomolar range, at least below about 50 nM, or below about 25 nM, or below about 15 nM, or below about 10 nM, or below about 5 nM, or within the range of about 0.05 nM or about 0.1 nM to about 50 nM.

[0054] In some embodiments, the peptide compound of formula (I) has a sequence selected from SEQ ID NO: 1 to 50, or their salts or solvates, as shown in Table 2 below.

[0055] In some embodiments, the peptide compound of formula (I) is expressed in IC50 at a concentration of about 50 nM or less, or about 25 nM or less, or about 15 nM or less, or about 10 nM or less. 50 Inhibit IL-23-induced STAT3 phosphorylation.

[0056] In some embodiments, the peptide compound of formula (I) has a stability of about 20%, or about 15%, or about 10% in water at a pH of about 1.2, which is measured as a loss of purity after 24 hours in a buffer solution at 37°C.

[0057] In some embodiments, the peptide compound of formula (I) has a stability of about 10%, or about 5%, or about 2% in water at a pH of about 6.5, as measured by a loss of purity after 24 hours in a buffer solution at 37°C.

[0058] In some embodiments, the peptide compound of formula (I) has a stability of about 10%, or about 5%, or about 2% in water at a pH of about 7.4, as measured by a loss of purity after 24 hours in a buffer solution at 37°C.

[0059] In some embodiments, the peptide compound of formula (I) has at least 50% protease stability, which is measured as the percentage of the remaining portion of the peptide in simulated intestinal fluid at 60 min and 37°C.

[0060] In some embodiments, the peptide compound of formula (I) is used to treat and / or prevent autoimmune or inflammatory diseases in subjects in need.

[0061] In some embodiments, the autoimmune or inflammatory disease is selected from inflammatory bowel diseases such as Crohn's disease and ulcerative colitis, psoriasis, psoriatic arthritis, and hidradenitis suppurativa.

[0062] In some embodiments, this disclosure relates to compositions comprising a peptide compound of formula (I) as described herein, or a salt or solvation thereof, mixed with a carrier. The carrier may be a pharmaceutically acceptable excipient or transporter.

[0063] In some embodiments, this disclosure relates to a pharmaceutical composition comprising at least one peptide compound of formula (I) or a pharmaceutically acceptable salt or solvate thereof and at least one pharmaceutically acceptable excipient.

[0064] This disclosure also relates to the use of peptide compounds of formula (I) as described herein as pharmaceuticals, particularly for the treatment of conditions as described herein.

[0065] In some embodiments, this disclosure relates to the use of peptide compounds of formula (I) in the preparation of medicaments for the treatment and / or prevention of autoimmune or inflammatory diseases in subjects of need.

[0066] In some embodiments, this disclosure relates to a method for treating and / or preventing autoimmune or inflammatory diseases, the method comprising at least the step of administering a peptide compound of formula (I) to the subject. Detailed Implementation definition

[0067] Unless otherwise defined herein, scientific and technical terms used in connection with this disclosure shall have the meanings commonly understood by one of ordinary skill in the art. For example, *Concise Dictionary of Biomedicine and Molecular Biology*, Juo, Pei-Show, 2nd edition, 2002, CRC Press; *The Dictionary of Cell and Molecular Biology*, 3rd edition, 1999, Academic Press; and *Oxford Dictionary of Biochemistry and Molecular Biology*, revised edition, 2000, Oxford University Press can provide a general dictionary for those skilled in the art of the use of many terms used in this disclosure. While similar or equivalent methods and materials may be used in the practice or testing of this disclosure, exemplary methods and materials are described below. In the event of any conflict, this specification (including definitions) shall prevail. Generally, the nomenclature and techniques used in conjunction with those described herein for cell and tissue culture, molecular biology, virology, immunology, microbiology, genetics, analytical chemistry, synthetic organic chemistry, medical and medicinal chemistry, and protein and nucleic acid chemistry and hybridization are those well-known and commonly used in the art. Enzymatic reactions and purification techniques are performed according to the manufacturer's instructions as commonly practiced in the art or as described herein. Furthermore, unless the context otherwise requires, singular terms should include plural terms, and plural terms should include singular terms.

[0068] Units, prefixes, and symbols are all expressed in their SI-accepted form. Numerical ranges include the numbers that define the range. Unless otherwise stated, amino acid sequences are written from left to right, from amino to carboxyl. The headings provided herein are not intended to limit any aspect of this disclosure. Therefore, the terms defined below are defined more fully by referring to the specification (in its entirety).

[0069] Although this article cites many documents, this citation does not imply an admission that any of these documents constitutes part of the common knowledge in the field.

[0070] Throughout this specification and embodiments, the terms “have” and “comprise” or variations (such as “has”, “having”, “comprises”, or “comprising”) are used interchangeably and have the same meaning.

[0071] It should be noted that the terms "a" or "an" refer to one or more species; for example, "a peptide compound" should be understood to mean one or more peptide compounds. Therefore, the terms "a" (or "an"), "one or more species" and "at least one" are used interchangeably herein.

[0072] Furthermore, the use of "and / or" herein should be interpreted as explicitly disclosing each of the two specified features or components, with or without the other. Therefore, the term "and / or," as used herein in phrases such as "A and / or B," is intended to include "A and B," "A or B," "A" (alone), and "B" (alone). Similarly, the term "and / or," as used in phrases such as "A, B, and / or C," is intended to cover each of the following: A, B, and C; A, B, or C; A or C; A or B; B or C; A and C; A and B; B and C; A (alone); B (alone); and C (alone).

[0073] The term “about” or “approximately” is used herein to mean approximately, roughly, approximately, or around. When the term “about” is used in conjunction with a numerical range, it modifies the range by extending the upper and lower boundaries of the indicated value. Generally, the term “about” can define a value to vary above and below a specified value by, for example, 10% or more (higher or lower). In some embodiments, the term indicates a deviation from the indicated value of ±10%, ±5%, ±4%, ±3%, ±2%, ±1%, ±0.9%, ±0.8%, ±0.7%, ±0.6%, ±0.5%, ±0.4%, ±0.3%, ±0.2%, ±0.1%, ±0.05%, or ±0.01%. In some embodiments, “about” indicates a deviation from the indicated value of ±10%. In some embodiments, “about” indicates a deviation from the indicated value of ±5%. In some embodiments, “about” indicates a deviation from the indicated value of ±4%. In some embodiments, “about” indicates a deviation from the indicated value of ±3%. In some embodiments, “about” indicates a deviation from the indicated value of ±2%. In some embodiments, "about" indicates a deviation from the indicated value of ±1%. In some embodiments, "about" indicates a deviation from the indicated value of ±0.9%. In some embodiments, "about" indicates a deviation from the indicated value of ±0.8%. In some embodiments, "about" indicates a deviation from the indicated value of ±0.7%. In some embodiments, "about" indicates a deviation from the indicated value of ±0.6%. In some embodiments, "about" indicates a deviation from the indicated value of ±0.5%. In some embodiments, "about" indicates a deviation from the indicated value of ±0.4%. In some embodiments, "about" indicates a deviation from the indicated value of ±0.3%. In some embodiments, "about" indicates a deviation from the indicated value of ±0.1%. In some embodiments, "about" indicates a deviation from the indicated value of ±0.05%. In some embodiments, "about" indicates a deviation from the indicated value of ±0.01%.

[0074] Peptide compounds. The term "peptide compound" encompasses both the singular and plural "peptide compound" and refers to a molecule containing amino acids linked by amide bonds. The peptide compounds disclosed herein contain native and non-native amino acids and structural units such as butyl moieties. The peptides disclosed herein contain amide and alkyl bonds based on native and non-native amino acids, and structural units involved in these bonds. The peptide compounds disclosed herein are cyclized. Peptide compounds may contain additional modifications using functionalized amino acids.

[0075] Amino acids. Amino acids in this article are represented by their names, their commonly known three-letter symbols, or by the single-letter symbols recommended by the IUPAC-IUB Committee on Biochemical Terminology. Therefore, the amino acid sequences disclosed herein include the standard single-letter and three-letter codes for naturally occurring amino acids, as well as the generally accepted three-letter codes for other non-natural amino acids, such as Aib for α-aminoisobutyric acid, Orn for ornithine, or Ahx for 6-aminohexanoic acid.

[0076] As used herein, the term "amino acid" or "any amino acid" means any and all amino acids, including naturally occurring amino acids, modified amino acids, and non-natural amino acids. It includes both D-amino acids and L-amino acids. Natural amino acids include those found in nature, such as the 23 amino acids that, for example, combine into peptide chains to form the structural units of a wide range of proteins. These are primarily L-stereomers, although some D-amino acids are found in bacterial envelopes and some antibiotics. Unless otherwise stated, L-amino acids are used throughout this disclosure. D-amino acids are indicated as such.

[0077] Residue. In this disclosure, the term "residue" refers to a portion of a peptide compound that is formed by the combination of an amino acid or structural unit with another amino acid or structural unit through the formation of an amide bond. A residue is i) the portion remaining after the removal of a water element from an amino acid, ii) the portion remaining after the removal of an OH group from a dicarboxylic acid, and iii) the portion remaining after the removal of one of the H groups from a structural unit having a single amino group.

[0078] Amide bond. The term "amide bond" refers to a bond obtained through coupling between the amine moiety of an amino acid of the peptide compound disclosed herein and the carboxylic acid moiety of another amino acid of the peptide compound. For example, an amide bond can be obtained between the amine moiety of X2 (Aud, Ade, Add, Atd, Atea, Ahd, or Aoa) and the carboxyl moiety of the side chain of X9 (Glu). Alternatively, an amide bond can be obtained between the carboxyl moiety of X2 (Dec, Dod, Oca, or Dpa) and the amine moiety of the side chain of X9 (Lys).

[0079] Activity. The term “activity” as used with respect to peptide compounds refers to the ability of a peptide compound to inhibit the interleukin-23 receptor. As used herein, the term “activity” also refers to the ability of a peptide compound to inhibit intracellular STAT3 phosphorylation induced by activation of the IL-23 receptor.

[0080] Potency. "Potency" is a measure of a compound's ability to inhibit the activation of the interleukin-23 receptor, measured in cell-based assays, such as those shown in the examples. Numerically, it can be expressed as "IC". 50The "value" is the effective concentration of a compound that, in a dose-response experiment, inhibits the response by a maximum increase in the half-maximum (e.g., phosphorylation of STAT3 induced by IL-23 receptor activation).

[0081] The compounds disclosed herein have interleukin-23 receptor affinity. This term refers to the ability to bind to the interleukin-23 receptor. The interleukin-23 receptor affinity or activity of the peptide compounds disclosed herein can be tested using the assays described in the Methods section, and the results are shown in Tables 8 or 9 herein.

[0082] Inhibition of the binding of interleukin-23 to its receptor. The statement "inhibition of the binding of interleukin-23 to its receptor" refers to the property of the disclosed peptide compounds to prevent or reduce the binding of the IL-23 receptor to its ligand IL-23, thereby preventing or reducing the activation of downstream cellular responses induced by the binding of IL-23 to its receptor. The ability of the peptide compounds to inhibit the binding of IL-23 to IL-23R is measured by the assays disclosed in the Examples section. This property is measured as binding affinity and can be expressed as "IC50". 50 The value is the effective concentration of the compound that induces a maximum half-maximal reduction in the binding of IL-2.3 to its receptor in a dose-response experiment. In some embodiments, the peptide compounds disclosed herein have a binding affinity to the interleukin-23 receptor of 100 nM or less (i.e., IC50). 50 ≤ 100 nM).

[0083] The terms "pharmaceutically acceptable solvate" and "pharmaceutically acceptable salt" refer to solvates or salts of the peptide compounds disclosed herein that are suitable for formulation of pharmaceutical compositions and are physiologically acceptable, i.e., safe and effective for use in mammals. Pharmaceutically acceptable salts or solvates are suitable for oral administration.

[0084] As used herein, “administer” means to deliver the peptide compound or composition described herein to a subject. The peptide compound or composition may be administered to a subject using methods known in the art. In particular, the peptide compound or composition may be administered orally, sublingually, buccally, nasally, rectally, vaginally, or via the lungs, or again intravenously, subcutaneously, intramuscularly, or intradermally. In some embodiments, administration is oral.

[0085] Table 1 presents the codes for the amino acids and other structural units used in this paper: Table 1: Natural and Non-natural Amino Acids and Structural Units

[0086] It should be understood that, for clarity, certain features of this disclosure described in the context of individual embodiments may also be provided in combination in a single embodiment. Conversely, for brevity, multiple features of this disclosure described in the context of a single embodiment may also be provided individually or in any suitable sub-combination.

[0087] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure pertains. All publications mentioned herein are incorporated herein by reference to disclose and describe methods and / or materials relating to the cited publications.

[0088] A list of sources, ingredients and components as described below is provided, so that combinations and mixtures thereof are also conceived and are within the scope of this document.

[0089] It should be understood that each maximum numerical limit given throughout this specification includes each lower numerical limit, as if such lower numerical limit were explicitly stated herein. Each minimum numerical limit given throughout this specification will include each higher numerical limit, as if such higher numerical limit were explicitly stated herein. Each numerical range given throughout this specification will include each narrower numerical range falling within such a wider range, as if such narrower numerical range were explicitly stated in its entirety herein.

[0090] All item lists (such as ingredient lists) include their combinations and mixtures.

[0091] The references herein may include trade names of components, including the various ingredients used in this disclosure. Materials equivalent to those mentioned under trade names (e.g., materials obtained from different sources under different names or reference numbers) may be substituted and used in the description herein. peptide compounds

[0092] In some embodiments, this disclosure relates to a peptide compound that is capable of binding to the IL-23 receptor (IL23-R) and inhibiting or reducing the binding of interleukin-23 (IL-23) to its receptor.

[0093] The peptide compounds disclosed herein are peptide compounds of formula (I):

[0094] X1(§1)(#1)-X2(#1)(@2)-X3-X4-X5-X6-X7-X8-X9(§1)(@2)-X10-X11-X12-X13-R1(I),

[0095] in

[0096] X1 is absent or has a butyl portion and

[0097] When X1 is a butyl group, then (§1) represents the alkyl bond between X1 and X9, and (#1) represents the alkyl bond between X1 and X2, and in such a case, (@2) does not exist, or

[0098] When X1 is absent, then (@2) represents the amide bond between X2 and X9, and in such a case, (§1) and (#1) are absent;

[0099] X2 is an amino acid residue selected from the following: Add, Ade, Ahd, Aoa, Atd, Atea, Aud, Dad, Dae, Dec, Dod, Dpa, Kme, Kme(αAc), Lys(αAc), Nmapt, Nmha, and Oca.

[0100] X3 is an amino acid residue selected from the following: Nak, Trp, Wdm, Wim, Wme, Wcl, and Wfl;

[0101] X4 is an amino acid residue selected from the following: Aib, Gln, Glu, Ile, Iva, Leu, Lys, Lys(Ac), Mle, Mly, Mva, Phe, Thr, Trp, and Val;

[0102] X5 is an amino acid residue selected from the following: Aib, Mkdm, Mle, and Thp;

[0103] X6 is an amino acid residue selected from the following: Trp, Tyr, Yae, Yde, and Yme;

[0104] X7 is an amino acid residue selected from the following: Nak, Nal, and Trp;

[0105] X8 is an amino acid residue selected from the following: Aib, Cba, Cit, Gln, Leu, Lys, Lys(Ac), Mle, Mly, D-Mkdm, Mkdm, Thp, D-Trp, and D-Tza;

[0106] X9 is an amino acid residue selected from the following: Asp, Glu, Kme, Lys, and Orn;

[0107] X10 is an amino acid residue selected from the following: Ala, Asn, Gly, and Ser;

[0108] X11 is an amino acid residue selected from the following: Ala, Asn, Bal, Gly, Hol, Hph, His, Ile, Iva, Leu, Mhis, Pal, Pyal, PyEA, Val, and their corresponding D-forms;

[0109] X12 is absent or consists of amino acid residues selected from the following: Bal, Dnmy, Ile, Lys, Mep, Mys, and Sar;

[0110] X13 is absent or is a Lys residue; and...

[0111] R1 either does not exist or is selected from -NH2, -OH and -N(C2H5)2;

[0112] Or its salts or solvates.

[0113] The expression "and its corresponding D-form" necessarily refers to amino acid residues that contain an asymmetric carbon atom and exist in both L and D forms. This expression does not apply to amino acids that do not contain such an asymmetric carbon atom, such as glycine (Gly) and PyEA.

[0114] The peptide compound of formula (I) inhibits the binding of interleukin-23 (IL-23) to its receptor (IL-23R).

[0115] When used with respect to the peptide compounds disclosed herein, the term "solvent" refers to an aggregate consisting of a peptide compound and one or more solvent molecules (e.g., organic solvent molecules and / or water). Such solvates may originate from the solvent used for crystallization, be inherent in the solvent used for preparation or crystallization, or be foreign to such solvents. Such solvates are within the scope of this disclosure.

[0116] When referring to the peptide compounds disclosed herein, the term "salt" is intended to mean the ionic form of the peptide compounds disclosed herein, which is negatively or positively charged and interacts ionicly with positively charged cations or negatively charged anions. Such salts include acid addition salts or base addition salts formed with inorganic acids or with organic acids or bases.

[0117] In some embodiments, X1 is a butyl moiety, and X9 is an amino acid residue selected from the following: Kme, Lys, and Orn.

[0118] In some embodiments, X1 is the butyl portion and X9 represents Kme.

[0119] In some embodiments, X1 is absent, X2 is an amino acid residue selected from the following: Dad, Dae, Dec, Dod, Dpa, and Oca, and X9 is an amino acid residue selected from the following: Kme, Lys, and Orn.

[0120] In some embodiments, X1 is absent, X2 is an amino acid residue selected from the following: Dec, Dod, Dpa, and Oca, and X9 is an amino acid residue selected from the following: Kme, Lys, and Orn.

[0121] In some embodiments, X1 is absent, X2 is an amino acid residue selected from Dec, Dod, Dpa and Oca, and X9 is an amino acid residue selected from Kme and Lys.

[0122] In some embodiments, X1 is absent, X2 is an amino acid residue selected from the following: Add, Ade, Ahd, Aoa, Atd, Atea, Aud, Kme, Kme(αAc), Lys(αAc), Nmapt, and Nmha, and X9 is an amino acid residue selected from the following: Asp and Glu.

[0123] In some embodiments, X1 is absent, X2 is an amino acid residue selected from Add, Ade, Ahd, Aoa, Atd, Atea, and Aud, and X9 is Glu.

[0124] In some embodiments, X1 is a butyl moiety, and X9 is a residue of Kme, or X1 is absent, X2 is an amino acid residue selected from the following: Dad, Dae, Dec, Dod, Dpa, and Oca, and X9 is an amino acid residue selected from the residues Kme, Lys, and Orn, or X1 is absent, X2 is an amino acid residue selected from the following: Dec, Dod, Dpa, and Oca, and X9 is an amino acid residue selected from the following: Kme and Lys, or X1 is absent, X2 is an amino acid residue selected from the following: Add, Ade, Ahd, Aoa, Atd, Atea, Aud, Kme, Kme(αAc), Lys(αAc), Nmapt, and Nmha, and X9 is an amino acid residue selected from the following: Asp and Glu, or X1 does not exist, X2 is an amino acid residue selected from the following: Add, Ade, Ahd, Aoa, Atd, Atea, and Aud, and X9 is a residue of Glu.

[0125] In some embodiments, the peptide compound of formula (I) comprises a first group of compounds, wherein:

[0126] X1 is absent or is a butyl portion;

[0127] X2 is an amino acid residue selected from the following: Add, Ade, Ahd, Aoa, Atd, Atea, Aud, Dec, Dod, Dpa, Kme, Kme(αAc), Nmapt, Nmha, and Oca;

[0128] X3 is an amino acid residue selected from the following: Trp, Wdm, Wim, and Wme;

[0129] X4 is an amino acid residue selected from the following: Gln, Leu, and Lys(Ac);

[0130] X5 is an amino acid residue selected from the following: Aib and Thp;

[0131] X6 is a residue of Yde;

[0132] X7 is a residue of Nal;

[0133] X8 is an amino acid residue selected from the following: Aib, Mkdm, and Thp;

[0134] X9 is an amino acid residue selected from the following: Glu, Kme, and Lys;

[0135] X10 is an amino acid residue selected from the following: Asn, Gly, and Ser;

[0136] X11 is an amino acid residue selected from the following: Pal and PyEA;

[0137] X12 is absent or is a residue of Sar;

[0138] X13 does not exist;

[0139] R1 is absent or -NH2.

[0140] Or its salts or solvates.

[0141] In some embodiments, the peptide compound of formula (I) comprises a first group of compounds, wherein:

[0142] X2 is an amino acid residue selected from the following: Add, Ade, Ahd, Aoa, Atd, Atea, Aud, Dec, Dod, Dpa, Kme, Kme(αAc), Nmapt, Nmha, and Oca;

[0143] X3 is an amino acid residue selected from the following: Trp, Wdm, Wim, and Wme;

[0144] X4 is an amino acid residue selected from the following: Gln, Leu, and Lys(Ac);

[0145] X5 is an amino acid residue selected from the following: Aib and Thp;

[0146] X6 is a residue of Yde;

[0147] X7 is a residue of Nal;

[0148] X8 is an amino acid residue selected from the following: Aib, Mkdm, and Thp;

[0149] X9 is an amino acid residue selected from the following: Glu, Kme, and Lys;

[0150] X10 is an amino acid residue selected from the following: Asn, Gly, and Ser;

[0151] X11 is an amino acid residue selected from the following: Pal and PyEA;

[0152] X12 is absent or is a residue of Sar;

[0153] X13 does not exist; and...

[0154] R1 is either absent or -NH2.

[0156] In some embodiments of the first group, the peptide compound of formula (I) is expressed at an IC50 concentration of less than about 50 nM, or less than about 25 nM, or less than about 15 nM, or less than about 10 nM. 50 Inhibit IL-23-induced STAT3 phosphorylation.

[0157] In some examples of peptide compounds of the first formula (I):

[0158] X10 is a residue of Asn, or

[0159] X10 is an amino acid residue selected from the following: Gly and Ser, and X12 is Sar.

[0160] The peptide compounds of such embodiments have an IC50 of about 25 nM or less, or about 0.05 nM to about 25 nM, for inhibiting IL-23-induced STAT3 phosphorylation. 50 .

[0161] In some embodiments of the peptide compounds of the first formula (I), R1 is NH2.

[0162] The peptide compounds of such embodiments have an IC50 of about 10 nM or less, or about 0.05 nM to about 10 nM, for inhibiting IL-23-induced STAT3 phosphorylation. 50 .

[0163] In some examples of peptide compounds of the first formula (I):

[0164] R1 is NH2, and

[0165] When X1 is present, X2 is an amino acid residue selected from the following: Kme, Nmapt, and Nmha; or

[0166] When X1 is absent, X2 is an amino acid residue selected from the following: Add, Ade, Aoa, Atd, Atea, Aud, Dec, Dod, and Oca.

[0167] In some examples of peptide compounds of the first formula (I):

[0168] X10 represents Asn; or,

[0169] X10 represents an amino acid residue selected from the following: Gly and Ser, and X12 represents Sar; and

[0170] R1 represents NH2, or

[0171] R1 represents NH2 and

[0172] When X1 is a butyl moiety, X2 represents an amino acid residue selected from the following: Kme, Nmapt, and Nmha; or

[0173] When X1 is absent, X2 represents an amino acid residue selected from Add, Ade, Aoa, Atd, Atea, Aud, Dec, Dod, and Oca.

[0175] The peptide compounds of such embodiments have an IC50 of about 5 nM or less, or about 0.05 nM to about 5 nM, for inhibiting IL-23-induced STAT3 phosphorylation. 50 .

[0176] In some embodiments, X1 is absent or is a butyl portion.

[0177] In some embodiments, X1 does not exist.

[0178] In some embodiments, X1 is the butyl portion.

[0179] In some embodiments, X2 is an amino acid residue selected from the following: Add, Ade, Ahd, Aoa, Atd, Atea, Aud, Dad, Dae, Dec, Dod, Dpa, Kme, Kme(αAc), Lys(αAc), Nmapt, Nmha, and Oca.

[0180] In some embodiments, X2 is an amino acid residue selected from the following: Add, Ade, Ahd, Aoa, Atd, Atea, Aud, Dec, Dod, Dpa, Kme, Kme(αAc), Nmapt, Nmha, and Oca.

[0181] In some embodiments, X2 is an amino acid residue selected from the following: Add, Ade, Ahd, Aoa, Atd, Atea, Aud, Dec, Dod, Kme, Kme(αAc), Nmapt, Nmha, and Oca.

[0182] In some embodiments, X2 is an amino acid residue selected from the following: Add, Ade, Aoa, Atd, Atea, Aud, Dec, Dod, Kme, Kme(αAc), Nmapt, Nmha, and Oca.

[0183] In some embodiments, X2 is a residue of Add.

[0184] In some embodiments, X2 is a residue of Ade.

[0185] In some embodiments, X2 is a residue of Ahd.

[0186] In some embodiments, X2 is a residue of Aoa.

[0187] In some embodiments, X2 is a residue of Atd.

[0188] In some embodiments, X2 is a residue of Atea.

[0189] In some embodiments, X2 is a residue of Aud.

[0190] In some embodiments, X2 is a residue of Dad.

[0191] In some embodiments, X2 is a residue of Dae.

[0192] In some embodiments, X2 is a residue of Dec.

[0193] In some embodiments, X2 is a residue of Dod.

[0194] In some embodiments, X2 is a residue of Dpa.

[0195] In some embodiments, X2 is a residue of Kme.

[0196] In some embodiments, X2 is a residue of Kme(αAc).

[0197] In some embodiments, X2 is a residue of Nmapt.

[0198] In some embodiments, X2 is a residue of Nmha.

[0199] In some embodiments, X2 is a residue of Oca.

[0200] In some embodiments, X3 is an amino acid residue selected from the following: Nak, Trp, Wdm, Wim, Wme, Wcl, and Wfl.

[0201] In some embodiments, X3 is an amino acid residue selected from the following: Trp, Wdm, Wim, and Wme.

[0202] In some embodiments, X3 is an amino acid residue selected from the following: Trp, Wim, and Wme.

[0203] In some embodiments, X3 is a residue of Trp.

[0204] In some embodiments, X3 is a residue of Wdm.

[0205] In some embodiments, X3 is a residue of Wim.

[0206] In some embodiments, X3 is a residue of Wme.

[0207] In some embodiments, X4 is an amino acid residue selected from the following: Aib, Gln, Glu, Ile, Iva, Leu, Lys, Lys(Ac), Mle, Mly, Mva, Phe, Thr, Trp, and Val.

[0208] In some embodiments, X4 is an amino acid residue selected from the following: Gln, Leu, and Lys(Ac).

[0209] In some embodiments, X4 is a residue of Gln.

[0210] In some embodiments, X4 is a residue of Leu.

[0211] In some embodiments, X4 is a residue of Lys(Ac).

[0212] In some embodiments, X5 is an amino acid residue selected from the following: Aib, Mkdm, Mle, and Thp.

[0213] In some embodiments, X5 is an amino acid residue selected from the following: Aib and Thp.

[0214] In some embodiments, X5 is a residue of Aib.

[0215] In some embodiments, X5 is a residue of Thp.

[0216] In some embodiments, X6 is an amino acid residue selected from the following: Trp, Tyr, Yae, Yde, and Yme.

[0217] In some embodiments, X6 is a residue of Yde.

[0218] In some embodiments, X7 is an amino acid residue selected from Nak, Nal, and Trp.

[0219] In some embodiments, X7 is a residue of Nal.

[0220] In some embodiments, X8 is an amino acid residue selected from the following: Aib, Cba, Cit, Gln, Leu, Lys, Lys(Ac), Mle, Mly, D-Mkdm, Mkdm, Thp, D-Trp, and D-Tza.

[0221] In some embodiments, X8 is an amino acid residue selected from the following: Aib, Mkdm, and Thp.

[0222] In some embodiments, X8 is a residue of Aib.

[0223] In some embodiments, X8 is a residue of Mkdm.

[0224] In some embodiments, X8 is a residue of Thp.

[0225] In some embodiments, X9 is an amino acid residue selected from the following: Asp, Glu, Kme, Lys, and Orn.

[0226] In some embodiments, X9 is an amino acid residue selected from the following: Glu, Kme, and Lys.

[0227] In some embodiments, X9 is a residue of Glu.

[0228] In some embodiments, X9 is a residue of Kme.

[0229] In some embodiments, X9 is a residue of Lys.

[0230] In some embodiments, X10 is an amino acid residue selected from the following: Ala, Asn, Gly, and Ser.

[0231] In some embodiments, X10 is an amino acid residue selected from the following: Asn, Gly, and Ser.

[0232] In some embodiments, X10 is a residue of Asn.

[0233] In some embodiments, X10 is a residue of Gly.

[0234] In some embodiments, X10 is a residue of Ser.

[0235] In some embodiments, X11 is an amino acid residue selected from the following: Ala, Asn, Bal, Gly, Hol, Hph, His, Ile, Iva, Leu, Mhis, Pal, Pyal, PyEA, Val, and their corresponding D-forms.

[0236] In some embodiments, X11 is an amino acid residue selected from the following: Ala, Asn, Bal, Gly, Hol, Hph, His, Ile, Iva, Leu, D-Leu, Mhis, Pal, Pyal, PyEA, and Val.

[0237] In some embodiments, X11 is an amino acid residue selected from Pal and PyEA.

[0238] In some embodiments, X11 is a residue of Pal.

[0239] In some embodiments, X11 is a residue of PyEA.

[0240] In some embodiments, X12 is absent or is an amino acid residue selected from the following: Bal, Dnmy, Ile, Lys, Mep, and Sar.

[0241] In some embodiments, X12 is absent or is a residue of Sar.

[0242] In some embodiments, X12 is not present.

[0243] In some embodiments, X12 is a residue of Sar.

[0244] In some embodiments, X13 is absent or is a residue of Lys.

[0245] In some embodiments, X13 is not present.

[0246] In some embodiments, R1 is absent or selected from residues of -NH2, -OH or -N(C2H5)2.

[0247] In some embodiments, R1 is absent or is -NH2.

[0248] In some embodiments, R1 does not exist.

[0249] In some embodiments, R1 is NH2.

[0250] In some embodiments, the peptide compound of formula (I) comprises a second group of compounds, wherein:

[0251] X1 is absent or is a butyl portion;

[0252] X2 is an amino acid residue selected from the following: Add, Ade, Ahd, Aoa, Atd, Atea, Aud, Dec, Dod, Dpa, Kme, Kme(αAc), Nmapt, Nmha, and Oca;

[0253] X3 is an amino acid residue selected from the following: Trp, Wim, and Wme;

[0254] X4 is an amino acid residue selected from the following: Gln, Leu, and Lys(Ac);

[0255] X5 is an amino acid residue selected from the following: Aib and Thp;

[0256] X6 is a residue of Yde;

[0257] X7 is a residue of Nal;

[0258] X8 is an amino acid residue selected from the following: Aib, Mkdm, and Thp;

[0259] X9 is an amino acid residue selected from the following: Glu, Kme, and Lys;

[0260] X10 is an amino acid residue selected from the following: Asn, Gly, and Ser;

[0261] X11 is an amino acid residue selected from the following: Pal and PyEA;

[0262] X12 is absent or is a residue of Sar;

[0263] X13 does not exist;

[0264] R1 is either absent or -NH2.

[0265] In some embodiments of the second group:

[0266] When X2 is a residue of Nmapt, X11 is a residue of PyEA, and X4 is a residue of Gln, then X10 is not a residue of Gly, or

[0267] When X2 is a residue of Nmapt, X11 is a residue of PyEA, and X10 is a residue of Gly, then X4 is not a residue of Gln.

[0268] When X2 is a residue of Nmha and X11 is a residue of PyEA, then X4 is not a residue of Lys(Ac).

[0269] In some embodiments of the second group, when X2 is a residue of Nmapt, then X11 is a residue of Pal.

[0270] In some embodiments of the second group, X10 is a Ser residue when X2 is a residue of Nmapt and X11 is a residue of PyEA.

[0271] In some embodiments of the second group, when X2 is a residue of Nmha, then X11 is a residue of Pal.

[0272] In some embodiments of the second group, X4 is a residue of Gln when X2 is a residue of Nmha and X11 is a residue of PyEA.

[0273] In some embodiments of the second group, the peptide compound of formula (I) has an IC50 of about 25 nM or less, or about 0.05 nM to about 25 nM, for inhibiting IL-23-induced STAT3 phosphorylation. 50 .

[0274] In some embodiments, the peptide compound of formula (I) comprises a third group of compounds, wherein:

[0275] X1 is absent or is a butyl portion;

[0276] X2 is an amino acid residue selected from the following: Add, Ade, Ahd, Aoa, Atd, Atea, Aud, Dec, Dod, Dpa, Kme, Kme(αAc), Nmapt, Nmha, and Oca;

[0277] X3 is an amino acid residue selected from the following: Trp, Wim, and Wme;

[0278] X4 is an amino acid residue selected from the following: Gln, Leu, and Lys(Ac);

[0279] X5 is an amino acid residue selected from the following: Aib and Thp;

[0280] X6 is a residue of Yde;

[0281] X7 is a residue of Nal;

[0282] X8 is an amino acid residue selected from the following: Aib, Mkdm, and Thp;

[0283] X9 is an amino acid residue selected from the following: Glu, Kme, and Lys;

[0284] X10 is an amino acid residue selected from the following: Asn, Gly, and Ser;

[0285] X11 is an amino acid residue selected from the following: Pal and PyEA;

[0286] X12 is absent or is a residue of Sar;

[0287] X13 does not exist;

[0288] R1 is either absent or -NH2.

[0289] In some embodiments of the third group,

[0290] When X2 is a residue of Nmapt, X11 is a residue of PyEA, and X4 is a residue of Gln, then X10 is not a residue of Gly, or

[0291] When X2 is a residue of Nmapt, X11 is a residue of PyEA, and X4 is a residue of Lys(Ac), then X10 is not a residue of Ser, or

[0292] When X2 is a residue of Nmapt, X11 is a residue of PyEA, and X10 is a residue of Gly, then X4 is not a residue of Gln.

[0293] When X2 is a residue of Nmha, then X11 is not a residue of PyEA.

[0294] In some embodiments of the third group, when X2 is a residue of Nmapt, then X11 is a residue of Pal.

[0295] In some embodiments, the peptide compound of formula (I) comprises a third group of compounds, wherein:

[0296] X2 is an amino acid residue selected from the following: Add, Ade, Ahd, Aoa, Atd, Atea, Aud, Dec, Dod, Dpa, Kme, Kme(αAc), Nmapt, Nmha, and Oca;

[0297] X3 is an amino acid residue selected from the following: Trp, Wim, and Wme;

[0298] X4 is an amino acid residue selected from the following: Gln, Leu, and Lys(Ac);

[0299] X5 is an amino acid residue selected from the following: Aib and Thp;

[0300] X6 is a residue of Yde;

[0301] X7 is a residue of Nal;

[0302] X8 is an amino acid residue selected from the following: Aib, Mkdm, and Thp;

[0303] X9 is an amino acid residue selected from the following: Glu, Kme, and Lys;

[0304] X10 is an amino acid residue selected from the following: Asn, Gly, and Ser;

[0305] X11 is an amino acid residue selected from the following: Pal and PyEA;

[0306] X12 is absent or is a residue of Sar;

[0307] X13 does not exist;

[0308] R1 is absent or -NH2.

[0309] in

[0310] When X2 is a residue of Nmapt, X11 is a residue of PyEA, and X4 is a residue of Gln, then X10 is not a residue of Gly, or

[0311] When X2 is a residue of Nmapt, X11 is a residue of PyEA, and X10 is a residue of Gly, then X4 is not a residue of Gln, or

[0312] When X2 is a residue of Nmha and X11 is a residue of PyEA, then X4 is not a residue of Lys(Ac).

[0314] In some embodiments of the third group, when X2 is a residue of Nmapt and X11 is a residue of PyEA, then X10 is a residue of Gly and X4 is a residue of Gln, or X10 is a residue of Ser and X4 is a residue of Lys(Ac).

[0315] In some embodiments of the third group, when X2 is a residue of Nmha, then X11 is a residue of Pal.

[0316] The third group of peptide compounds has an IC50 of about 15 nM or less, or about 0.05 nM to about 15 nM, for inhibiting IL-23-induced STAT3 phosphorylation. 50 .

[0317] In some embodiments, the peptide compound of formula (I) comprises a fourth group of compounds, wherein:

[0318] X1 is absent or is a butyl portion;

[0319] X2 is an amino acid residue selected from the following: Add, Ade, Ahd, Aoa, Atd, Atea, Aud, Dec, Dod, Kme, Kme(αAc), Nmapt, Nmha, and Oca;

[0320] X3 is an amino acid residue selected from the following: Trp, Wim, and Wme;

[0321] X4 is an amino acid residue selected from the following: Gln, Leu, and Lys(Ac);

[0322] X5 is an amino acid residue selected from the following: Aib and Thp;

[0323] X6 is a residue of Yde;

[0324] X7 is a residue of Nal;

[0325] X8 is an amino acid residue selected from the following: Aib, Mkdm, and Thp;

[0326] X9 is an amino acid residue selected from the following: Glu, Kme, and Lys;

[0327] X10 is an amino acid residue selected from the following: Asn, Gly, and Ser;

[0328] X11 is an amino acid residue selected from the following: Pal and PyEA;

[0329] X12 is absent or is a residue of Sar;

[0330] X13 does not exist;

[0331] R1 is absent or -NH 2，

[0332] In some embodiments of the fourth group,

[0333] When X2 is a residue of Nmapt, then X11 is not a residue of PyEA, or

[0334] When X2 is a residue of Nmapt and X11 is a residue of PyEA, then X3 is a residue of Wim, X4 is a residue of Gln, and X10 is a residue of Ser, or

[0335] When X2 is a residue of Nmha, then X11 is not a residue of PyEA, or

[0336] If X2 is a residue of Aoa or Dec, then X4 is not a residue of Leu.

[0337] In some embodiments of the fourth group, when X2 is a residue of Nmapt, then X11 is a residue of Pal.

[0338] In some embodiments of the fourth group, when X2 is a residue of Nmapt and X11 is a residue of PyEA, then X10 is a residue of Gly, X4 is a residue of Gln and X3 is a residue of Wim.

[0339] In some embodiments of the fourth group, when X2 is a residue of Nmha, then X11 is a residue of Pal.

[0340] In some embodiments of the fourth group, X4 is a Gln residue when X2 is an Aoa or Dec residue.

[0341] In some embodiments, the peptide compound of formula (I) comprises a fourth group of compounds, wherein:

[0342] X2 is an amino acid residue selected from the following: Add, Ade, Ahd, Aoa, Atd, Atea, Aud, Dec, Dod, Dpa, Kme, Kme(αAc), Nmapt, Nmha, and Oca;

[0343] X3 is an amino acid residue selected from the following: Trp, Wim, and Wme;

[0344] X4 is an amino acid residue selected from the following: Gln, Leu, and Lys(Ac);

[0345] X5 is an amino acid residue selected from the following: Aib and Thp;

[0346] X6 is a residue of Yde;

[0347] X7 is a residue of Nal;

[0348] X8 is an amino acid residue selected from the following: Aib, Mkdm, and Thp;

[0349] X9 is an amino acid residue selected from the following: Glu, Kme, and Lys;

[0350] X10 is an amino acid residue selected from the following: Asn, Gly, and Ser;

[0351] X11 is an amino acid residue selected from the following: Pal and PyEA;

[0352] X12 is absent or is a residue of Sar;

[0353] X13 does not exist; and...

[0354] R1 is absent or -NH 2；

[0355] in

[0356] When X2 is a residue of Nmapt, X11 is a residue of PyEA, and X4 is a residue of Gln, then X10 is not a residue of Gly, or

[0357] When X2 is a residue of Nmapt, X11 is a residue of PyEA, and X4 is a residue of Lys(Ac), then X10 is not a residue of Ser, or

[0358] When X2 is a residue of Nmapt, X11 is a residue of PyEA, and X10 is a residue of Gly, then X4 is not a residue of Gln, or

[0359] When X2 is a residue of Nmha, then X11 is not a residue of PyEA.

[0360] In some embodiments, the peptide compound of formula (I) comprises a fourth group of compounds, wherein:

[0361] X2 is an amino acid residue selected from the following: Add, Ade, Ahd, Aoa, Atd, Atea, Aud, Dec, Dod, Kme, Kme(αAc), Nmapt, Nmha, and Oca;

[0362] X3 is an amino acid residue selected from the following: Trp, Wim, and Wme;

[0363] X4 is an amino acid residue selected from the following: Gln, Leu, and Lys(Ac);

[0364] X5 is an amino acid residue selected from the following: Aib and Thp;

[0365] X6 is a residue of Yde;

[0366] X7 is a residue of Nal;

[0367] X8 is an amino acid residue selected from the following: Aib, Mkdm, and Thp;

[0368] X9 is an amino acid residue selected from the following: Glu, Kme, and Lys;

[0369] X10 is an amino acid residue selected from the following: Asn, Gly, and Ser;

[0370] X11 is an amino acid residue selected from the following: Pal and PyEA;

[0371] X12 is absent or is a residue of Sar;

[0372] X13 does not exist;

[0373] R1 is absent or -NH 2，

[0374] in

[0375] When X2 is a residue of Nmapt, then X11 is not PyEA, or

[0376] When X2 is a residue of Nmapt and X11 is a residue of PyEA, then X3 is a residue of Wim, X4 is a residue of Gln, and X10 is a residue of Ser, or

[0377] When X2 is a residue of Nmha, then X11 is not a residue of PyEA, or

[0378] If X2 is a residue of Aoa or Dec, then X4 is not a residue of Leu.

[0380] The peptide compounds in the fourth group have an IC50 of about 10 nM or less, or about 0.05 nM to about 10 nM, for inhibiting IL-23-induced STAT3 phosphorylation. 50 .

[0381] In some embodiments, the peptide compound of formula (I) comprises a fifth group of compounds, wherein:

[0382] X1 is absent or is a butyl portion;

[0383] X2 is an amino acid residue selected from the following: Add, Ade, Aoa, Atd, Atea, Aud, Dec, Dod, Kme, Kme(αAc), Nmapt, Nmha, and Oca;

[0384] X3 is an amino acid residue selected from the following: Trp, Wim, and Wme;

[0385] X4 is an amino acid residue selected from the following: Gln, Leu, and Lys(Ac);

[0386] X5 is an amino acid residue selected from the following: Aib and Thp;

[0387] X6 is a residue of Yde;

[0388] X7 is a residue of Nal;

[0389] X8 is an amino acid residue selected from the following: Aib, Mkdm, and Thp;

[0390] X9 is an amino acid residue selected from the following: Glu, Kme, and Lys;

[0391] X10 is an amino acid residue selected from the following: Asn, Gly, and Ser;

[0392] X11 is a residue of Pal;

[0393] X12 is a residue of Sar;

[0394] X13 does not exist;

[0395] R1 is -NH2.

[0396] In some embodiments of the fifth group,

[0397] When X2 is a residue of Nmapt and X3 is a residue of Wme, then X8 is not a residue of Mkdm, or

[0398] When X2 is a residue of Aoa, then X4 is not a residue of Leu.

[0399] In some embodiments of Group 5, X4 is a residue of Gln when X2 is a residue of Aoa or Dec.

[0400] In some embodiments, the peptide compound of formula (I) comprises a fifth group of compounds, wherein:

[0401] X2 is an amino acid residue selected from the following: Add, Ade, Aoa, Atd, Atea, Aud, Dec, Dod, Kme, Kme(αAc), Nmapt, Nmha, and Oca;

[0402] X3 is an amino acid residue selected from the following: Trp, Wim, and Wme;

[0403] X4 is an amino acid residue selected from the following: Gln, Leu, and Lys(Ac);

[0404] X5 is an amino acid residue selected from the following: Aib and Thp;

[0405] X6 is a residue of Yde;

[0406] X7 is a residue of Nal;

[0407] X8 is an amino acid residue selected from the following: Aib, Mkdm, and Thp;

[0408] X9 is an amino acid residue selected from the following: Glu, Kme, and Lys;

[0409] X10 is an amino acid residue selected from the following: Asn, Gly, and Ser;

[0410] X11 is a residue of Pal;

[0411] X12 is a residue of Sar;

[0412] X13 does not exist;

[0413] R1 is -NH2.

[0414] in,

[0415] When X2 is a residue of Nmapt and X3 is a residue of Wme, then X8 is not a residue of Mkdm, or

[0416] When X2 is a residue of Aoa, then X4 is not a residue of Leu.

[0418] The peptide compounds in Group 5 have an IC50 of about 5 nM or less, or about 0.05 nM to about 5 nM, for inhibiting IL-23-induced STAT3 phosphorylation. 50 .

[0419] In some embodiments, the peptide compound of formula (I) comprises a sixth group, wherein:

[0420] X1 is the butyl portion;

[0421] X2 is an amino acid residue selected from the following: Nmap and Nmha;

[0422] X3 is an amino acid residue selected from the following: Wdm, Wim, and Wme;

[0423] X4 is an amino acid residue selected from the following: Gln, Leu, and Lys(Ac);

[0424] X5 is a residue of Thp;

[0425] X6 is a residue of Yde;

[0426] X7 is a residue of Nal;

[0427] X8 is a residue of Aib;

[0428] X9 is a residue of Kme;

[0429] X10 is a residue of Gly;

[0430] X11 is a residue of PyEA;

[0431] X12 does not exist;

[0432] X13 does not exist;

[0433] R1 does not exist.

[0434] In some embodiments of the peptide compounds in Group 6, X4 is not a Gln residue when X2 is a residue of Nmha.

[0435] The peptide compounds in group 6 have an IC50 value for inhibiting IL-23-induced STAT3 phosphorylation in the range of about 25 nM to about 50 nM. 50 .

[0436] In some embodiments, the peptide compound of formula (I) comprises a seventh group, wherein:

[0437] X1 is the butyl portion;

[0438] X2 is an amino acid residue selected from the following: Nmapt and Nmha;

[0439] X3 is an amino acid residue selected from the following: Wim and Wme;

[0440] X4 is an amino acid residue selected from the following: Gln and Lys(Ac);

[0441] X5 is residue Thp;

[0442] X6 is a residue of Yde;

[0443] X7 is a residue of Nal;

[0444] X8 is a residue of Aib;

[0445] X9 is a residue of Kme;

[0446] X10 is an amino acid residue selected from the following: Gly and Ser;

[0447] X11 is a residue of PyEA;

[0448] X12 does not exist;

[0449] X13 does not exist;

[0450] R1 does not exist.

[0451] In some embodiments of Group 7, X4 is not a Lys(Ac) residue when X2 is a residue of Nmha, or X4 is not a Gln residue when X2 is a residue of Nmapt.

[0452] The peptide compounds in group seven have an IC50 value for inhibiting IL-23-induced STAT3 phosphorylation in the range of about 15 nM to about 25 nM. 50 .

[0453] In some embodiments, the peptide compound of formula (I) comprises an eighth group, wherein:

[0454] X1 is absent or is a butyl portion;

[0455] X2 is an amino acid residue selected from the following: Aoa, Dec, Dpa, and Nmapt;

[0456] X3 is an amino acid residue selected from the following: Wim and Wme;

[0457] X4 is an amino acid residue selected from the following: Gln, Leu, and Lys(Ac);

[0458] X5 is a residue of Thp;

[0459] X6 is a residue of Yde;

[0460] X7 is a residue of Nal;

[0461] X8 is an amino acid residue selected from the following: Aib and Thp;

[0462] X9 is an amino acid residue selected from the following: Glu, Kme, and Lys;

[0463] X10 is an amino acid residue selected from the following: Asn, Gly, and Ser;

[0464] X11 is an amino acid residue selected from the following: Pal and PyEA;

[0465] X12 is absent or is a residue of Sar;

[0466] X13 does not exist;

[0467] R1 is absent or is a -NH2 residue.

[0468] In some embodiments of the peptide compounds in Group 8, X4 is not a Gln residue when X2 is an Aoa residue, or X4 is not a Gln residue when X2 is a Dec residue, or X8 is a Thrp residue when X2 is an Nmapt residue, or X3 is a Wim residue, X10 is a Ser residue, and X11 is a PyEA residue.

[0469] The peptide compounds in Group 8 have an IC50 value for inhibiting IL-23-induced STAT3 phosphorylation in the range of about 10 nM to about 15 nM. 50 .

[0470] In some embodiments, the peptide compound of formula (I) comprises a ninth group, wherein:

[0471] X1 is absent or is a butyl portion;

[0472] X2 is an amino acid residue selected from the following: Ahd and Nmapt;

[0473] X3 is an amino acid residue selected from the following: Wim and Wme;

[0474] X4 is a residue of Gln;

[0475] X5 is a residue of Thp;

[0476] X6 is a residue of Yde;

[0477] X7 is a residue of Nal;

[0478] X8 is an amino acid residue selected from the following: Aib or Mkdm;

[0479] X9 is an amino acid residue selected from the following: Glu and Kme;

[0480] X10 is an amino acid residue selected from the following: Asn and Ser;

[0481] X11 is an amino acid residue selected from the following: Pal and PyEA;

[0482] X12 is absent or is a residue of Sar;

[0483] X13 does not exist;

[0484] R1 is either absent or -NH2.

[0485] In some embodiments of Group 9, when X2 is a residue of Nmpat, then X3 is a residue of Wim and X11 is a residue of PyEA, or X8 is a residue of Mkdm.

[0486] The peptide compounds in group nine have an IC50 value for inhibiting IL-23-induced STAT3 phosphorylation in the range of about 5 nM to about 10 nM. 50 .

[0487] In some embodiments, the peptide compound of formula (I) comprises a tenth group, wherein:

[0488] X1 is absent or is a butyl portion;

[0489] X2 is an amino acid residue selected from the following: Aoa, Nmapt, and Nmha;

[0490] X3 is an amino acid residue selected from the following: Trp, Wim, and Wme;

[0491] X4 is an amino acid residue selected from the following: Gln and Lys(Ac);

[0492] X5 is a residue of Thp;

[0493] X6 is a residue of Yde;

[0494] X7 is a residue of Nal;

[0495] X8 is a residue of Aib;

[0496] X9 is an amino acid residue selected from the following: Glu and Kme;

[0497] X10 is an amino acid residue selected from the following: Asn, Gly, and Ser;

[0498] X11 is a residue of Pal;

[0499] X12 is a residue of Sar;

[0500] X13 does not exist;

[0501] R1 is -NH2.

[0502] In some embodiments of Group 10, when X2 is a residue of Nmha, then X4 is not a residue of Gln and / or X10 is not a residue of Asn; or when X2 is a residue of Nmapt, then X3 is not a residue of Wim; or when X2 is a residue of Nampt and X3 is a residue of Wme, then X10 is not a residue of Ser.

[0503] The peptide compounds in group 10 have an IC50 value in the range of about 3 nM to about 5 nM for inhibiting IL-23-induced STAT3 phosphorylation. 50 .

[0504] In some embodiments, the peptide compound of formula (I) comprises an eleventh group, wherein

[0505] X1 is the butyl portion;

[0506] X2 is an amino acid residue selected from the following: Nmapt and Nmha;

[0507] X3 is an amino acid residue selected from the following: Trp, Wim, and Wme;

[0508] X4 is an amino acid residue selected from the following: Gln and Lys(Ac);

[0509] X5 is a residue of Aib;

[0510] X6 is a residue of Yde;

[0511] X7 is a residue of Nal;

[0512] X8 is an amino acid residue selected from the following: Aib and Thp;

[0513] X9 is a residue of Kme;

[0514] X10 is an amino acid residue selected from the following: Gly and Ser;

[0515] X11 is a residue of Pal;

[0516] X12 is a residue of Sar;

[0517] X13 does not exist;

[0518] R1 is -NH2.

[0519] In some embodiments of group eleven, when X2 is a residue of Nmapt, then X8 is a residue of Thrp or X3 is not a residue of W, or X3 is a residue of Wim and X10 is a residue of Gly, or X3 is a residue of Wme and X10 is a residue of Ser, or when X2 is a residue of Nmha, then X4 is not a residue of Lys(Ac).

[0520] The peptide compounds in group eleven have an IC50 value in the range of about 2 nM to about 3 nM for inhibiting IL-23-induced STAT3 phosphorylation. 50 .

[0521] In some embodiments, the peptide compound of formula (I) comprises a twelfth group, wherein:

[0522] X1 is absent or is a butyl portion;

[0523] X2 is an amino acid residue selected from the following: Ade, Atd, Atea, Aud, and Nmapt;

[0524] X3 is a residue of Wim;

[0525] X4 is an amino acid residue selected from the following: Gln, Leu, and Lys(Ac);

[0526] X5 is a residue of Thp;

[0527] X6 is a residue of Yde;

[0528] X7 is a residue of Nal;

[0529] X8 is an amino acid residue selected from the following: Aib and Mkdm;

[0530] X9 is an amino acid residue selected from the following: Glu and Kme;

[0531] X10 is an amino acid residue selected from the following: Asn and Ser;

[0532] X11 is a residue of Pal;

[0533] X12 is a residue of Sar;

[0534] X13 does not exist;

[0535] R1 is -NH2.

[0536] In some embodiments of Group 12, X4 is not a Gln residue when X2 is a residue of Nmapt, or X8 is not a residue of Aib, or X10 is not a residue of Asn, or X4 is not a Gln residue when X2 is a residue of Ade, or X4 is a Leu residue when X2 is a residue of Aud.

[0537] The peptide compounds in group 12 have an IC50 value in the range of about 1 nM to about 2 nM for inhibiting IL-23-induced STAT3 phosphorylation. 50 .

[0538] In some embodiments, the peptide compound of formula (I) comprises group thirteen, wherein:

[0539] X1 is absent or is a butyl portion;

[0540] X2 is an amino acid residue selected from the following: Add, Ade, Aud, Dec, Dod, and Oca;

[0541] X3 is an amino acid residue selected from the following: Wim and Wme;

[0542] X4 is an amino acid residue selected from the following: Gln and Lys(Ac);

[0543] X5 is an amino acid residue selected from the following: Aib and Thp;

[0544] X6 is a residue of Yde;

[0545] X7 is a residue of Nal;

[0546] X8 is an amino acid residue selected from the following: Aib, Mkdm, and Thp;

[0547] X9 is an amino acid residue selected from the following: Glu, Kme, and Lys;

[0548] X10 is a residue of Asn;

[0549] X11 is a residue of Pal;

[0550] X12 is a residue of Sar;

[0551] X13 does not exist;

[0552] R1 is -NH2.

[0553] In some embodiments of Group 13, X8 is not a residue Mkdm when X2 is a residue of Nmapt or Nmha.

[0554] The peptide compounds in group thirteen have an IC50 value for inhibiting IL-23-induced STAT3 phosphorylation in the range of about 0.05 nM to about 1 nM. 50 .

[0555] In some embodiments, the peptide compound of formula (I) is represented by a sequence selected from SEQ ID NO: 1-50, or their salts or solvates, as shown in Table 2 below.

[0556] In some embodiments, the peptide compound of formula (I) is represented by a sequence selected from SEQ ID NO: 1-22, 24, 26-48 and 50, or their salts or solvates.

[0557] In some embodiments, the peptide compound of formula (I) is represented by a sequence selected from SEQ ID NO: 1-21, 24, 26-36, 38-48 and 50, or their salts or solvates.

[0558] In some embodiments, the peptide compound of formula (I) is represented by a sequence selected from SEQ ID NO: 1-8, 10-14, 16, 18-21, 24, 26-36, 39-47 and 50, or their salts or solvates.

[0559] In some embodiments, the peptide compound of formula (I) is represented by a sequence selected from SEQ ID NO: 1-2, 4-8, 10-14, 16, 18-21, 24, 26-35, 39-44, 46-47 and 50, or their salts or solvates.

[0560] In some embodiments, the peptide compound of formula (I) is represented by a sequence selected from SEQ ID NO: 1-2, 4-5, 7-8, 10-14, 16, 18-21, 26, 28-29, 31-35, 39-40, 43-44, 46-47 and 50, or their salts or solvates.

[0561] In some embodiments, the peptide compound of formula (I) is represented by a sequence selected from SEQ ID NO: 1-2, 4-5, 7-8, 10-14, 16, 18-21, 31-35, 39, 44, 46-47 and 50, or their salts or solvates.

[0562] In some embodiments, the peptide compound of formula (I) is represented by a sequence selected from SEQ ID NO: 4, 11, 16, 18, 19 and 50, or their salts or solvates. In some embodiments, the peptide compound of formula (I) is represented by a sequence selected from the following: -SEQ ID NO: 1-22, 24, 26-48 and 50, or their salts or solvates; or -SEQ ID NO: 1-21, 24, 26-36, 38-48 and 50, or their salts or solvates; or -SEQ ID NO: 1-8, 10-14, 16, 18-21, 24, 26-36, 39-47 and 50, or their salts or solvates; or -SEQ ID NO: 1-2, 4-8, 10-14, 16, 18-21, 24, 26-35, 39-44, 46-47 and 50, or their salts or solvates; or -SEQ ID NO: 1-2, 4-5, 7-8, 10-14, 16, 18-21, 26, 28-29, 31-35, 39-40, 43-44, 46-47 and 50, or their salts or solvates; or -SEQ ID NO: 1-2, 4-5, 7-8, 10-14, 16, 18-21, 31-35, 39, 44, 46-47 and 50, or their salts or solvates; or -SEQ ID NO: 4, 11, 16, 18, 19 and 50, or their salts or solvates.

[0563] Table 2: Sequences of peptide compounds SEQ ID NO: 1 to 50

[0564] (§1) represents the alkyl bond between X1 and X9, and (#1) represents the alkyl bond between X1 and X2.

[0565] (@2) represents the amide bond between X2 and X9.

[0566] (Ac) indicates the acetylation of an amino acid residue, such as Lys(Ac) or Kme(αAc).

[0567] The disclosed peptide compounds exhibit affinity for the interleukin-23 receptor and the ability to prevent or reduce the binding of IL-23 to its receptor.

[0568] The peptide compounds disclosed herein bind to interleukin-23 to prevent, reduce, or inhibit intracellular signal transduction pathways that lead to Th17 cell activation.

[0569] The assays described in the Examples section can be used to test the interleukin-23 receptor affinity or inhibitory activity of the peptide compounds disclosed herein, and the results are shown in Tables 8 and 9 herein.

[0570] The peptide compound of formula (I) determined using the method described in the examples has a binding affinity of 100 nM or less to the interleukin-23 receptor (i.e., human ELISA IL23 / IL23R) (i.e., IC50). 50 <= 100 nM), or 50 nM or smaller (i.e., IC 50 <= 50.0 nM), or 20 nM or less (i.e., IC 50 <= 20.0 nM), or 10 nM or smaller (i.e., IC 50 <=10.0 nM), or 5 nM or smaller (i.e., IC 50 <= 5.0 nM), or 1 nM or smaller (i.e., IC 50 <= 1.0 nM).

[0571] In some embodiments, the peptide compound of formula (I) is expressed in IC50 of about 50 nM or less, or about 25 nM or less, or about 15 nM or less, or about 10 nM or less, or about 5 nM or less, or about 3 nM or less, or about 2 nM or less, or about 1 nM or less. 50 Inhibit IL-23-induced STAT3 phosphorylation.

[0572] In some embodiments, the peptide compound of formula (I) is expressed in an IC50 concentration ranging from about 0.01 nM to about 50 nM, or from about 0.05 nM to about 50 nM, or from about 0.1 nM to about 50 nM. 50 Inhibit IL-23-induced STAT3 phosphorylation.

[0573] In some embodiments, the peptide compound of formula (I) is expressed in an IC50 concentration ranging from about 0.01 nM to about 25 nM, or from about 0.05 nM to about 25 nM, or from about 0.1 nM to about 25 nM. 50 Inhibit IL-23-induced STAT3 phosphorylation.

[0574] In some embodiments, the peptide compound of formula (I) is expressed in IC50 values ​​ranging from about 0.05 nM to about 50 nM, or about 0.05 nM to about 25 nM, or about 0.05 nM to about 15 nM, or about 0.05 nM to about 10 nM, or about 0.05 nM to about 5 nM, or about 0.05 nM to about 3 nM, or about 0.05 nM to about 2 nM, or about 0.05 nM to about 1 nM. 50 Inhibit IL-23-induced STAT3 phosphorylation.

[0575] In some embodiments, the peptide compound of formula (I) is expressed in IC50 values ​​ranging from about 0.1 nM to about 50 nM, or from about 0.1 nM to about 25 nM, or from about 0.1 nM to about 15 nM, or from about 0.1 nM to about 10 nM, or from about 0.1 nM to about 5 nM, or from about 0.1 nM to about 3 nM, or from about 0.1 nM to about 2 nM, or from about 0.1 nM to about 1 nM. 50 Inhibit IL-23-induced STAT3 phosphorylation.

[0576] In some embodiments, the peptide compound of formula (I) is expressed at an IC50 concentration in the range of about 0.01 nM to about 50 nM. 50 Inhibit IL-23-induced STAT3 phosphorylation.

[0577] In some embodiments, the peptide compound of formula (I) is expressed at an IC50 concentration in the range of about 0.01 nM to about 25 nM. 50 Inhibit IL-23-induced STAT3 phosphorylation.

[0578] In some embodiments, the peptide compound of formula (I) is expressed at an IC50 concentration in the range of about 0.01 nM to about 20 nM. 50 Inhibit IL-23-induced STAT3 phosphorylation.

[0579] In some embodiments, the peptide compound of formula (I) is expressed at an IC50 concentration in the range of about 0.01 nM to about 15 nM. 50 Inhibit IL-23-induced STAT3 phosphorylation.

[0580] In some embodiments, the peptide compound of formula (I) is expressed at an IC50 concentration in the range of about 0.01 nM to about 10 nM. 50 Inhibit IL-23-induced STAT3 phosphorylation.

[0581] In some embodiments, the peptide compound of formula (I) is expressed at an IC50 concentration in the range of about 0.01 nM to about 8 nM. 50 Inhibit IL-23-induced STAT3 phosphorylation.

[0582] In some embodiments, the peptide compound of formula (I) is expressed at an IC50 concentration in the range of about 0.01 nM to about 6 nM. 50 Inhibit IL-23-induced STAT3 phosphorylation.

[0583] In some embodiments, the peptide compound of formula (I) is expressed at an IC50 concentration in the range of about 0.01 nM to about 5 nM. 50 Inhibit IL-23-induced STAT3 phosphorylation.

[0584] In some embodiments, the peptide compound of formula (I) is expressed at an IC50 concentration in the range of about 0.01 nM to about 4 nM. 50 Inhibit IL-23-induced STAT3 phosphorylation.

[0585] In some embodiments, the peptide compound of formula (I) is expressed at an IC50 concentration in the range of about 0.01 nM to about 3 nM. 50 Inhibit IL-23-induced STAT3 phosphorylation.

[0586] In some embodiments, the peptide compound of formula (I) is expressed at an IC50 concentration in the range of about 0.01 nM to about 2 nM. 50 Inhibit IL-23-induced STAT3 phosphorylation.

[0587] In some embodiments, the peptide compound of formula (I) is expressed at an IC50 concentration in the range of about 0.01 nM to about 1 nM. 50 Inhibit IL-23-induced STAT3 phosphorylation.

[0588] In another embodiment, the peptide compound of formula (I) is sufficiently chemically stable in aqueous solution at different pH values. Stability in aqueous solution was measured as the loss of purity after 24 hours in a buffer solution at 37°C.

[0589] In some embodiments, the peptide compound of formula (I) has chemical stability in aqueous solution in a pH range of about 1 to about 8.5, or about 1.2 to about 7.4, as illustrated in the examples.

[0590] In some embodiments, the peptide compound of formula (I) is stable in aqueous solution at pH values ​​of about 1, 1.2, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.4, 7.5, 8, 8.5 or 9.

[0591] In some embodiments, the peptide compound of formula (I) is stable in aqueous solution at pH 1.2, 6.5 or 7.4.

[0592] As described in the Examples section, the stability of the peptide compound of formula (I) is determined by measuring the relative purity loss.

[0593] In some embodiments, the peptide compound of formula (I) has a stability in water of about 20% or less, or about 15% or less, or about 10% or less at a pH of about 1.2, which is measured as a loss of purity at 37°C for 24 hours after the test relative to a control at 37°C for 0 hours.

[0594] In some embodiments, the peptide compound of formula (I) has a stability of about 20% or less in water at a pH of about 1.2, which is measured as a loss of purity at 37°C for 24 hours after the test compared to a control at 37°C for 0 hours.

[0595] The peptide compound of formula (I) has a stability of about 15% or less in water at a pH of about 1.2, which was measured as a loss of purity after 24 hours at 37°C.

[0596] The peptide compound of formula (I) has a stability of about 10% or less in water at a pH of about 1.2, which was measured as a loss of purity at 37°C for 24 hours after the test compared with the control at 37°C for 0 hours.

[0597] The peptide compound of formula (I) has a stability of about 10% or less, or about 5% or less, or about 2% or less in water at a pH of about 6.5, which was measured as a loss of purity at 37°C for 24 hours after the test relative to the control at 37°C for 0 hours.

[0598] The peptide compound of formula (I) has a stability of about 10% or less in water at a pH of about 6.5, which was measured as a loss of purity at 37°C for 24 hours after the test compared to the control at 37°C for 0 hours.

[0599] The peptide compound of formula (I) has a stability of about 5% or less in water at a pH of about 6.5, which was measured as a loss of purity at 37°C for 24 hours after the test compared with the control at 37°C for 0 hours.

[0600] The peptide compound of formula (I) has a stability of about 2% or less in water at a pH of about 6.5, which was measured as a loss of purity at 37°C for 24 hours after the test compared to the control at 37°C for 0 hours.

[0601] The peptide compound of formula (I) has a stability of about 10% or less, or about 5% or less, or about 2% or less in water at a pH of about 7.4, which was measured as a loss of purity at 37°C for 24 hours after the test compared with the control at 37°C for 0 hours.

[0602] The peptide compound of formula (I) has a stability of about 10% or less in water at a pH of about 7.4, which was measured as a loss of purity at 37°C for 24 hours after the test compared with the control at 37°C for 0 hours.

[0603] The peptide compound of formula (I) has a stability of about 5% or less in water at a pH of about 7.4, which was measured as a loss of purity at 37°C for 24 hours after the test compared with the control at 37°C for 0 hours.

[0604] The peptide compound of formula (I) has a stability of about 2% or less in water at a pH of about 7.4, which was measured as a loss of purity at 37°C for 24 hours after the test compared with the control at 37°C for 0 hours.

[0605] In another embodiment, the peptide compound of formula (I) is sufficiently stable in the intestinal environment to exert its pharmacological effects.

[0606] Protease stability was measured as the percentage of the remaining peptide compound in simulated intestinal fluid at 60 min and 37°C, which was at least 50%.

[0607] The peptide compound of formula (I) has about 50% to about 100% protease stability, which is measured as the percentage of the remaining portion of the peptide in simulated intestinal fluid at 60 min and 37°C.

[0608] The peptide compound of formula (I) has about 60% to about 95% protease stability, which is measured as the percentage of the remaining portion of the peptide in simulated intestinal fluid at 37°C for 60 min.

[0609] The peptide compound of formula (I) has about 70% to about 90% protease stability, which is measured as the percentage of the remaining portion of the peptide in simulated intestinal fluid at 37°C for 60 min.

[0610] In some embodiments, the peptide compounds of formula (I) possess sufficient stability in the gastric or intestinal environment to exert their pharmacological effects. The stability of the peptides of formula (I) in the gastric or intestinal environment is determined as described in the Examples section. peptide synthesis

[0611] Technicians are familiar with a variety of different methods for preparing peptides. These methods include, but are not limited to, synthetic methods. Therefore, one method for preparing peptides is to synthesize them in solution or on a solid support, followed by separation and purification.

[0612] The method for preparing the compounds disclosed herein involves solid-phase synthesis on a suitable resin. Solid-phase peptide synthesis is a well-established method (see, for example: Stewart and Young, Solid Phase Peptide Synthesis, Pierce Chemical Co., Rockford, Ill., 1984; E. Atherton and R.C. Sheppard, Solid Phase Peptide Synthesis. A Practical Approach, Oxford-IRL Press, New York, 1989). Solid-phase synthesis is initiated by attaching the carboxyl terminus of an N-terminus protected amino acid to an inert solid support with a cleavable linker. This solid support can be any polymer that allows coupling of the initiating amino acid, such as triphenylmethyl resin, chlorotriphenylmethyl resin, Wang resin, or Rink resin, wherein the linkage of the carboxyl group (or the carboxamide of the Rink resin) to the resin is acid-sensitive (when using the Fmoc strategy). During peptide synthesis, the polymer support must remain stable under the conditions used to deprotect the α-amino groups. Alternatively, the Fmoc synthesis strategy can be applied, and Rink resin can be used. Another option is to apply the Fmoc synthesis strategy, and Wang resin can be used.

[0613] After the first protected amino acid at the N-terminus is coupled to a solid support, the α-amino protecting group of that amino acid is removed. The remaining protected amino acids are then coupled one after another in sequence, in peptide order, or to a pre-formed dipeptide, tripeptide, or tetrapeptide using a suitable amide coupling agent, such as BOP, HBTU, HATU, or DIC (N,N'-diisopropylcarbodiimide) / HOBt (1-hydroxybenzotriazole). BOP, HBTU, and HATU are used with a tertiary amine base. Alternatively, the released N-terminus can be functionalized with groups other than amino acids, such as carboxylic acids.

[0614] Typically, the reactive side chain groups of amino acids are protected with suitable blocking groups. These protecting groups are removed after the desired peptide has been assembled. They are removed under the same conditions as the desired product is cleaved from the resin. For information on protecting groups and the procedure for introducing protecting groups, see Protective Groups in Organic Synthesis, 3rd Edition, Greene, TW and Wuts, PGM, Wiley & Sons (New York: 1999).

[0615] In some cases, it may be necessary to have a selectively removable side-chain protecting group while other side-chain protecting groups remain intact. In such cases, the released functional groups can be selectively functionalized. For example, lysine is protected with an ivDde ([1-(4,4-dimethyl-2,6-dioxocyclohexyl-1-ylene)-3-methylbutyl) protecting group (SRChhabra et al., Tetrahedron Lett. [Tetrahedron Letters] 39, (1998), 1603), which is unstable to highly nucleophilic bases, such as 4% hydrazine in DMF (dimethylformamide). Therefore, if the N-terminal amino group and all side-chain functional groups are protected with acid-labile protecting groups, the ivDde group can be selectively removed using 4% hydrazine in DMF, and the corresponding free amino group can then be further modified, for example, by acylation. Alternatively, lysine can be coupled to a protected amino acid, and then the amino group of that amino acid can be deprotected to produce another free amino group, which can be acylated or attached to another amino acid.

[0616] Finally, the peptides are cleaved from the resin. This can be achieved using similar cleavage mixtures known to those skilled in the art, such as King's cocktail (DS King, CG Fields, GB Fields, Int. J. Peptide Protein Res. [International Journal of Peptide and Protein Research] 36, 1990, 255-266). For example, EDT can be replaced with DODT, or a mixture of TIS, water, and TFA can be used. The raw material can then be purified by chromatography (e.g., preparative RP-HPLC) if necessary.

[0617] All starting materials (such as amino acids and chemicals) can be ordered from suppliers or synthesized using methods known in the literature. Cycloning / lactam formation

[0618] Methods for forming lactam bonds are known to those skilled in the art. Exemplary methods are given in the examples.

[0619] In another respect, cyclization / lactam formation can be carried out after cutting from the resin.

[0620] In another aspect, cyclization / lactam formation can be performed after the peptide is cleaved from the resin and subsequently purified by methods known to those skilled in the art, such as preparative HPLC (in DMF).

[0621] In one aspect of cyclization, an amide bond is formed between a primary or secondary amine group of one amino acid and a carboxylic acid group of another amino acid. The carboxylic acid group can be activated with PyBOP or HATU in the presence of DIPEA to form a lactam with an unprotected amine (such as compounds having SEQ ID NO. 1-12, 14-21, 35, 50).

[0622] In another aspect of cyclization, the primary or secondary amine group of one amino acid can be covalently linked to the primary or secondary amine group of another amino acid. Cyclization can be carried out using bis(2,5-dioxopyrrolidone-1-yl) succinates (such as compounds having SEQ ID NO. 13, 22-34, 36-49).

[0623] On the other hand, cleavage from the Wang resin followed by cyclization with bis(2,5-dioxopyrrolidone-1-yl)succinate results in a free carboxylic acid C-terminus. This residue can be activated with HATU in the presence of DIPEA, and the addition of 2-(3-pyridyl)ethylamine (PyEA) yields a peptide with PyEa as the terminal group. Therapeutic uses

[0624] Another aspect of this disclosure relates to peptide compounds of formula (I) for the treatment and / or prevention of autoimmune or inflammatory diseases.

[0625] The peptide compound of formula (I) is intended for use by subjects in need. In this document, the terms “subject,” “individual,” and “patient” are used interchangeably. In some exemplary embodiments, an individual or subject is a person.

[0626] This disclosure also relates to a method for treating autoimmune or inflammatory diseases, the method comprising at least the step of administering a peptide compound of formula (I) to a patient in need.

[0627] This disclosure also relates to the use of peptide compounds of formula (I) in the preparation of medicaments for the treatment of autoimmune or inflammatory diseases.

[0628] The terms “disease” and “disorder” are used interchangeably and are intended to refer to any pathological or unhealthy state. In particular, a disease or disorder involves autoimmune or inflammatory diseases.

[0629] "Treatment" means administering a compound or composition, or a combination of compounds or compositions, to a subject in order to: eliminate a disease or disorder; prevent or slow down a subject's disease or disorder; inhibit or slow down the development of a new disease or disorder in a subject; reduce the frequency or severity of symptoms and / or relapses in a subject who currently has or previously had a disease or disorder; and / or prolong, i.e., increase the subject's lifespan. In particular, the term "treating / treatment" of a disease or disorder includes curing, shortening the duration, improving, slowing down, or inhibiting the progression or worsening of a disease or disorder or its symptoms.

[0630] "Preventing" specifically refers to administering a compound or composition, or a combination of compounds or compositions, to a subject in order to suppress or delay the onset of a disease or disorder in the subject.

[0631] In some embodiments, this disclosure includes methods for inhibiting IL-23 signaling by using cells expressing the IL-23 receptor, methods comprising contacting the cells with a peptide compound of formula (I). In some embodiments, the cells are mammalian cells. In some embodiments, the method is performed in vitro or in vivo. In some embodiments, inhibition of IL-23 signaling can be determined by measuring changes in phosphate-STAT3 levels in the cells.

[0632] In some embodiments, inhibition of IL-23 binding to IL-23R occurs in specific organs or tissues of the subject, such as the stomach, small intestine, large intestine / colon, intestinal mucosa, lamina propria, Peyer's patches, mesenteric lymph nodes, or lymphatic ducts.

[0633] Further, the use of a peptide compound of formula (I) or a composition comprising a compound of formula (I) in the preparation of a pharmaceutical composition for treating an autoimmune disease or an inflammatory disease is provided.

[0634] Autoimmune or inflammatory diseases can be, for example, inflammatory bowel diseases such as Crohn's disease and ulcerative colitis, psoriasis, psoriatic arthritis, and hidradenitis suppurativa.

[0635] In some embodiments, the disease or disorder is an autoimmune inflammatory condition and related diseases and disorders, such as multiple sclerosis, asthma, rheumatoid arthritis, inflammatory bowel disease (IBD), juvenile IBD, adolescent IBD, Crohn's disease, sarcoidosis, systemic lupus erythematosus, ankylosing spondylitis (axial spondyloarthritis), psoriatic arthritis, or psoriasis. In some embodiments, the disease or disorder is psoriasis (e.g., plaque psoriasis, guttate psoriasis, inverted psoriasis, pustular psoriasis, palmoplantar pustulosis, psoriasis vulgaris, or erythrodermic psoriasis), atopic dermatitis, atopic acne, ulcerative colitis, Crohn's disease, celiac disease (non-tropical stomatitis), enteropathy associated with seroreactive arthropathy, microscopic colitis, collagenous colitis, eosinophilic gastroenteritis / esophagitis, colitis associated with radiotherapy or chemotherapy, colitis associated with congenital immune disorders (such as leukocyte adhesion defect-1), chronic granulomatous disease, type 1 glycogen storage disease, Hermansky-Pudlak syndrome, Chediak-Higashi syndrome, or Wiskott-Aldrich syndrome. Syndrome), colitis following rectal and colectomy and ileoanal anastomosis, gastrointestinal cancer, pancreatitis, insulin-dependent diabetes mellitus, mastitis, cholecystitis, cholangitis, primary biliary cirrhosis, virus-associated enteropathy, pericholangitis, chronic bronchitis, chronic sinusitis, asthma, uveitis, or graft-versus-host disease. Pharmaceutical Composition

[0636] According to another aspect, a pharmaceutical composition comprising a peptide compound of formula (I) as described herein, or a salt or solvation thereof, as an active ingredient is disclosed. The pharmaceutical composition disclosed herein comprises at least one peptide compound of formula (I) as disclosed herein, or a pharmaceutically acceptable salt or solvation thereof, and at least one pharmaceutically acceptable excipient.

[0637] Depending on the desired form of the drug and the method of administration, the excipient is selected from commonly used excipients known to those skilled in the art.

[0638] Standard acceptable drug carriers and their formulations are known to those skilled in the art and are described, for example, in: Remington: The Science and Practice of Pharmacy, (20th edition) edited by AR Gennaro AR, 2000, Lippencott Williams & Wilkins and RCRowe et al. (eds.), Handbook of Pharmaceutical Excipients, PhP, updated May 2013.

[0639] In these pharmaceutical compositions intended for oral, sublingual, subcutaneous, intramuscular, intravenous, topical, local, intratracheal, intranasal, transdermal, or rectal administration, the active ingredient of formula (I) or a salt or solvation thereof may be administered to humans in unit doses with a mixture of conventional pharmaceutical excipients for the prevention or treatment of disorders or diseases or conditions at least partially affected by IL-23 receptor-induced reduction or inhibition of intracellular signaling. The desired effect may be the prevention, reduction, or treatment of inflammatory or autoimmune diseases, or at least one symptom of such diseases. Application unit

[0640] Appropriate unit forms of administration include oral forms such as tablets, soft or hard gel capsules, powders, granules, and oral solutions or suspensions; sublingual, buccal, intratracheal, intraocular, and intranasal forms; forms for inhalation, local, percutaneous, subcutaneous, intramuscular, or intravenous administration; rectal forms; and implants.

[0641] When prepared in unit-dosage form, the pharmaceutical compositions disclosed herein typically contain 0.01 mg to 1000 mg of a peptide compound of formula (I) described herein, or a salt or solvate thereof, as the active ingredient. The amount of the active ingredient combined with one or more excipients to produce a single unit-dosage form will have to vary depending on the subject of treatment and the specific route of administration. For example, formulations intended for oral administration to humans will generally contain, for example, 0.01 mg to 0.5 g of the active ingredient, along with an appropriate and convenient amount of excipients, which may vary from about 5 percent to about 98 percent by weight of the total composition. dose

[0642] The dose of the peptide compound of formula (I) described herein, or its salt or solvate, which is the active ingredient, can be administered orally at a dose of about 0.01 to 50 mg / dose, or 0.02 to 1 mg / dose, or for example, 0.0001 to 300 mg / kg body weight per day or 1 to 300 mg / kg body weight per day, either once or in divided doses.

[0643] There may be specific circumstances where higher or lower doses are appropriate; such doses do not depart from the scope of this disclosure. As is customary, the appropriate dose for each patient is determined by a physician based on the administration method and the patient's weight and response. [Example]

[0644] The following examples illustrate embodiments of the present disclosure that are currently known. However, it should be understood that the following are merely examples or illustrations of the application of the principles of the present disclosure. Many modifications and alternative compositions, methods, and systems have been devised by those skilled in the art without departing from the spirit and scope of the present disclosure. Therefore, while the present disclosure has been specifically described above, the following examples provide further details relating to what is now considered to be the most practical and preferred embodiments of the present disclosure. The abbreviations used are as follows:

[0645] AA: Amino acid

[0646] ACN: Acetonitrile

[0647] Aib: α-Amino-isobutyric acid

[0648] Boc: tert-butoxycarbonyl

[0649] tBu: tert-butyl

[0650] DCM: Dichloromethane

[0651] DIC: N,N'-Diisopropylcarbodiimide

[0652] DIPEA: N,N-Diisopropylethylamine

[0653] DMF: Dimethylformamide

[0654] DODT: 3,6-dioxa-1,8-octanedithiol

[0655] DPBS: Dulbecco phosphate-buffered saline

[0656] EDC: 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide

[0657] EDT: Ethylene dithiol

[0658] Fmoc: fluorenylmethoxycarbonyl

[0659] g: grams

[0660] HATU: O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethylureonium hexafluorophosphate

[0661] HBTU: 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyl-ureonium hexafluorophosphate

[0662] HOAt: 1-Hydroxy-7-azabenzotriazole

[0663] HOBt: 1-Hydroxybenzotriazole

[0664] HPLC: High Performance Liquid Chromatography

[0665] LC / MS: Liquid Chromatography / Mass Spectrometry

[0666] mM: millimoles

[0667] MMT: Monomethoxy-triphenylmethyl

[0668] na: Not available

[0669] nd: Undetermined

[0670] nM: Nanomolar

[0671] NMPN-methyl-2-pyrrolidone

[0672] PBS: Phosphate-buffered saline

[0673] Pfp: 2,2,4,6,7-pentamethyldihydrobenzofuran-5-sulfonyl

[0674] PyBOP Benzotriazine-1-yl-oxytripyrrolidinephosphide hexafluorophosphate

[0675] tBu: tert-butyl

[0676] TFA: Trifluoroacetic acid

[0677] Trt: Triphenylmethyl / Triphenylmethyl

[0678] UHPLC: Ultra-high pressure liquid chromatography

[0679] UV: Ultraviolet Material

[0680] Different rink-amide resins (e.g., 4-(2',4'-dimethoxyphenyl-Fmoc-aminomethyl)-phenoxyacetamido-leucylaminomethyl resin, MERCK BIOSCIENCES; 4-[(2,4-dimethoxyphenyl)(Fmoc-amino)methyl]phenoxyacetamidomethyl resin, Agilent Technologies) were used to synthesize peptide amides with loadings in the range of 0.2–0.7 mmol / g.

[0681] The natural amino acids protected by Fmoc were purchased from Protein Technologies Inc., Senn Chemicals, Merck Biosciences, Novabiochem, Iris Biotech, Bachem, Chem-Impex International, or Matrick Innovation. The following standard amino acids were used throughout the synthesis: Fmoc-L-Ala-OH, Fmoc-Arg(Pbf)-OH, Fmoc-L-Asn(Trt)-OH, Fmoc-L-Asp(OtBu)-OH, Fmoc-L-Gln(Trt)-OH, Fmoc-L-Glu(OtBu)-OH, Fmoc-Gly-OH, Fmoc-L-Leu-OH, Fmoc-L-Lys(Boc)-OH, Fmoc-L-Phe-OH, Fmoc-L-Ser-OH, and Fmoc-L-Trp(Boc)-OH.

[0682] In addition, the following specific amino acids were purchased from the same supplier as mentioned above: Fmoc-Aib-OH, Fmoc-L-Nal-OH, Fmoc-L-Phe(2-Ae-Boc)-OH, Fmoc-L-Yde-OH, Fmoc-L-Phe(4-Ome)-OH, Fmoc-L-Trp(Me)-OH, Fmoc-L-alpha-Me-Leu-OH, Fmoc-beta(4-thiazolyl)-Ala-OH, Fmoc-4-amino-tetrahydropyran-4-carboxylic acid, Fmoc -L-Sar-OH, Fmoc-L-Lys(Ac)-OH, Fmoc-L-Lys(Boc)Me-OH, Fmoc-L-Pal-OH, Fmoc-Ahx-OH, Fmoc-Apt-OH, Fmoc-Atd-OH , Fmoc-Add-OH, Fmoc-Ahd-OH, Fmoc-Aud-OH, Fmoc-Ade-OH, Fmoc-Aoa-OH, Fmoc-Atea-OH, Fmoc-Nmha-OH and Fmoc-Nmapt.

[0683] In addition, the following structural units were purchased from the same suppliers as those mentioned above: Fmoc-L-1-Nal-OH, Fmoc-L-Phe(2-Ae-Boc)-OH, Fmoc-L-Phe(Me)-OH, Fmoc-L-Orn(Boc), Fmoc-L-Wim-OH, Fmoc-L-Wme-OH, Fmoc-L-Wdm-OH, and Fmoc-PyEA.

[0684] In addition, the following structural units were purchased from the same supplier as described above: Dae, Dad, Dod, Dec, Dpa, and Oca as disuccinimid esters. Example 1: General Synthesis of Peptide Compounds Mkdm Synthesis

[0685] 470 mg of (2S)-6-(tert-butoxycarbonylamino)-2-(9H-fluorene-9-ylmethoxycarbonylamino)hexanoic acid was treated with 5 mL of TFA for two hours. Then, 100 mL of water was added, and the solution was frozen and lyophilized. The residue was absorbed with 10 mL of methanol. Then, 129 mg of DIPEA, 126 mg of NaCNBH3, and 50 mg of formaldehyde were added, and the mixture was stirred for one hour. The product was purified by chromatography to a yield of 230 mg. Yde Synthesis

[0686] Yae protected by BOC / Fmoc was treated with 95% TFA for 2 hours. Then, 1 mmol of Yae with the Fmoc protecting group attached to an α-amine was dissolved in a 1:1 mixture of MeOH and water. The pH was adjusted to 6–7 with DIPEA. Formaldehyde (3 mmol) and NaCNBH3 (2 mmol) were then added, and the mixture was stirred at room temperature for 1 hour. The mixture was concentrated and lyophilized. The resulting oil was purified by chromatography to give Yde. Synthesis of peptide compounds

[0687] Solid-phase peptide synthesis is performed on a PRELUDE peptide synthesizer (Protein Technologies) or a similar automated synthesizer using standard Fmoc chemistry and HBTU / DIPEA or HATU / DIPEA activation. DMF is used as the solvent.

[0688] Deprotection: 20% piperidine / DMF, for 2 × 2.5 min.

[0689] Wash: 7 × DMF.

[0690] Coupling: 5 / 5 / 13 200 mM AA / 500 mM HBTU in DMF / 2M DIPEA in NMP 2x for 20 min. Wash: 5 × DMF.

[0691] All standard couplings are activated using HBTU / DIPEA.

[0692] For N-terminal acetylated peptides, the N-terminal Fmoc protecting group was removed, and the peptides were treated twice with 10% acetic anhydride and DIPEA solution for 20 minutes under oscillation.

[0693] The peptides were cleaved from the resin using either King's cleavage mixture (82.5% TFA, 5% phenol, 5% water, 5% anisole, and 2.5% EDT) or a modified cleavage mixture (82.5% TFA, 5% phenol, 5% water, 5% anisole, and 2.5% DODT). The crude peptides were then precipitated in diethyl ether or diisopropyl ether, centrifuged, and lyophilized. The peptides were analyzed by analytical HPLC and examined by ESI mass spectrometry. The crude peptides were purified using a standard preparative RP-HPLC purification procedure. Synthesis of peptide SEQ. ID NO: 4

[0694] Aud(@2)-Wim-Q-Thp-Yde-Nal-Aib-E(@2)-N-Pal-Sar-NH2

[0695] Peptide SEQ ID NO: 4 was prepared using FMOC solid-phase peptide synthesis technology.

[0696] The RINK amide AM resin LL was used, with the synthesis conditions protected by a Prelude synthesizer and the standard Fmoc as reported in the literature.

[0697] Resin preparation: 0.1 mmol RINK amide AM resin LL (0.345 g, 0.29 mmol / g loading) was transferred to a 25 ml peptide container with a filter glass frit. The resin was washed three times with 4 ml DMF for 5 min each time. Step I: Coupling of Fmoc-2-(methylamino)acetic acid (Sar):

[0698] To deprotect the resin-bound Fmoc groups, add 20% piperidine from 4 ml of DMF to the swollen resin and mix by bubbling with nitrogen for 2.5 min, then purge. Repeat this process once.

[0699] After deprotection, the resin was washed 7 times with 4 ml DMF under mixed conditions. A solution of Fmoc-2-(methylamino)acetic acid (2.5 equivalents, 2.5 ml, 100 mM, in DMF) was added to the resin, followed by HATU (5 equivalents, 1 ml, 500 mM, in DMF) and DIPEA (13 equivalents, 0.65 ml, 2 M, in NMP). The mixture was stirred by bubbling with nitrogen for 40 min, then purged. The resin was washed once with 4 ml DMF, and the coupling process was repeated once. Before starting the next deprotection-coupling cycle, the resin was washed 5 times with 4 ml DMF by bubbling with N2. Step II: Coupling of Fmoc-(2S)-2-amino-3-(3-pyridyl)propionic acid (Pal):

[0700] To deprotect the resin-bound Fmoc groups, add 20% piperidine from 4 ml of DMF to the swollen resin and mix by bubbling with nitrogen for 2.5 min, then purge. Repeat this process once.

[0701] After deprotection, the resin was washed 7 times with 4 ml DMF under mixed conditions. A solution of Fmoc-(2S)-2-amino-3-(3-pyridyl)propionic acid (2.5 equivalents, 2.5 ml, 100 mM, in DMF) was added to the resin, followed by the addition of HATU (5 equivalents, 1 ml, 500 mM, in DMF) and DIPEA (13 equivalents, 0.65 ml, 2 M, in NMP). The mixture was stirred by bubbling with nitrogen for 40 min and then purged. The resin was washed once with 4 ml DMF, and the coupling process was repeated once. Before starting the next deprotection-coupling cycle, the resin was washed 5 times with 4 ml DMF by bubbling with N2. Step III: Coupling of Fmoc-Asn(Trt)-OH(N):

[0702] To deprotect the resin-bound Fmoc groups, add 20% piperidine from 4 ml of DMF to the swollen resin and mix by bubbling with nitrogen for 2.5 min, then purge. Repeat this process once.

[0703] After deprotection, the resin was washed 7 times with 4 ml DMF under mixed conditions. A solution of Fmoc-Asn(Trt)-OH (5 equivalents, 2.5 ml, 200 mM, in DMF) was added to the resin, followed by the addition of HATU (5 equivalents, 1 ml, 500 mM, in DMF) and DIPEA (13 equivalents, 0.65 ml, 2 M, in NMP). The mixture was stirred by bubbling with nitrogen for 40 min, then purged. The resin was washed once with 4 ml DMF, and the coupling process was repeated once. Before starting the next deprotection-coupling cycle, the resin was washed 5 times with 4 ml DMF by bubbling with N2. Step IV: Coupling of Fmoc-Asp(OtBu)-OH(E):

[0704] To deprotect the resin-bound Fmoc groups, add 20% piperidine from 4 ml of DMF to the swollen resin and mix by bubbling with nitrogen for 2.5 min, then purge. Repeat this process once.

[0705] After deprotection, the resin was washed 7 times with 4 ml DMF under mixed conditions. A solution of Fmoc-Asp(OtBu)-OH (5 equivalents, 2.5 ml, 200 mM, in DMF) was added to the resin, followed by the addition of HATU (5 equivalents, 1 ml, 500 mM, in DMF) and DIPEA (13 equivalents, 0.65 ml, 2 M, in NMP). The mixture was stirred by bubbling with nitrogen for 40 min, then purged. The resin was washed once with 4 ml DMF, and the coupling process was repeated once. Before starting the next deprotection-coupling cycle, the resin was washed 5 times with 4 ml DMF by bubbling with N2. Step V: Coupling of Fmoc-aminoisobutyric acid (Aib):

[0706] To deprotect the resin-bound Fmoc groups, add 20% piperidine from 4 ml of DMF to the swollen resin and mix by bubbling with nitrogen for 2.5 min, then purge. Repeat this process once.

[0707] After deprotection, the resin was washed 7 times with 4 ml DMF under mixed conditions. A solution of Fmoc-aminoisobutyric acid (2.5 equivalents, 2.5 ml, 100 mM, in DMF) was added to the resin, followed by HATU (5 equivalents, 1 ml, 500 mM, in DMF) and DIPEA (13 equivalents, 0.65 ml, 2 M, in NMP). The mixture was stirred by bubbling with nitrogen for 40 min, then purged. The resin was washed once with 4 ml DMF, and the coupling process was repeated once. Before starting the next deprotection-coupling cycle, the resin was washed 5 times with 4 ml DMF by bubbling with N2. Step VI: Coupling of Fmoc-(2S)-2-amino-3-(2-naphthyl)propionic acid (Nal):

[0708] To deprotect the resin-bound Fmoc groups, add 20% piperidine from 4 ml of DMF to the swollen resin and mix by bubbling with nitrogen for 2.5 min, then purge. Repeat this process once.

[0709] After deprotection, the resin was washed 7 times with 4 ml DMF under mixed conditions. A solution of Fmoc-(2S)-2-amino-3-(2-naphthyl)propionic acid (2.5 equivalents, 2.5 ml, 100 mM, in DMF) was added to the resin, followed by the addition of HATU (5 equivalents, 1 ml, 500 mM, in DMF) and DIPEA (13 equivalents, 0.65 ml, 2 M, in NMP). The mixture was stirred by bubbling with nitrogen for 40 min and then purged. The resin was washed once with 4 ml DMF, and the coupling process was repeated once. Before starting the next deprotection-coupling cycle, the resin was washed 5 times with 4 ml DMF by bubbling with N2. Step VII: Fmoc-(2S)-2-amino-3-[4-[2-(dimethylamino)ethoxy]phenyl]propionic acid (Yde) Couplet:

[0710] To deprotect the resin-bound Fmoc groups, add 20% piperidine from 4 ml of DMF to the swollen resin and mix by bubbling with nitrogen for 2.5 min, then purge. Repeat this process once.

[0711] After deprotection, the resin was washed 7 times with 4 ml DMF under mixed conditions. A solution of Fmoc-(2S)-2-amino-3-[4-[2-(dimethylamino)ethoxy]phenyl]propionic acid (2.5 equivalents, 2.5 ml, 100 mM, in DMF) was added to the resin, followed by the addition of HATU (5 equivalents, 1 ml, 500 mM, in DMF) and DIPEA (13 equivalents, 0.65 ml, 2 M, in NMP). The mixture was stirred by bubbling with nitrogen for 40 min and then purged. The resin was washed once with 4 ml DMF, and the coupling process was repeated once. Before starting the next deprotection-coupling cycle, the resin was washed 5 times with 4 ml DMF by bubbling with N2. Step VIII: Coupling of Fmoc-4-aminotetrahydropyran-4-carboxylic acid (Thp):

[0712] To deprotect the resin-bound Fmoc groups, add 20% piperidine from 4 ml of DMF to the swollen resin and mix by bubbling with nitrogen for 2.5 min, then purge. Repeat this process once.

[0713] After deprotection, the resin was washed 7 times with 4 ml DMF under mixed conditions. A solution of Fmoc-4-aminotetrahydropyran-4-carboxylic acid (2.5 equivalents, 2.5 ml, 100 mM, in DMF) was added to the resin, followed by the addition of HATU (5 equivalents, 1 ml, 500 mM, in DMF) and DIPEA (13 equivalents, 0.65 ml, 2 M, in NMP). The mixture was stirred by bubbling with nitrogen for 40 min, then purged. The resin was washed once with 4 ml DMF, and the coupling process was repeated once. Before starting the next deprotection-coupling cycle, the resin was washed 5 times with 4 ml DMF by bubbling with N2. Step IX: Coupling of Fmoc-Gln(Trt)-OH(Q):

[0714] To deprotect the resin-bound Fmoc groups, add 20% piperidine from 4 ml of DMF to the swollen resin and mix by bubbling with nitrogen for 2.5 min, then purge. Repeat this process once.

[0715] After deprotection, the resin was washed 7 times with 4 ml DMF under mixed conditions. A solution of Fmoc-Gln(Trt)-OH (5 equivalents, 2.5 ml, 200 mM, in DMF) was added to the resin, followed by the addition of HATU (5 equivalents, 1 ml, 500 mM, in DMF) and DIPEA (13 equivalents, 0.65 ml, 2 M, in NMP). The mixture was stirred by bubbling with nitrogen for 40 min, then purged. The resin was washed once with 4 ml DMF, and the coupling process was repeated once. Before starting the next deprotection-coupling cycle, the resin was washed 5 times with 4 ml DMF by bubbling with N2. Step X: Coupling of Fmoc-(2S)-2-amino-3-(7-methyl-1H-indole-3-yl)propionic acid (Wim):

[0716] To deprotect the resin-bound Fmoc groups, add 20% piperidine from 4 ml of DMF to the swollen resin and mix by bubbling with nitrogen for 2.5 min, then purge. Repeat this process once.

[0717] After deprotection, the resin was washed seven times with 4 ml DMF under mixed conditions. A solution of Fmoc-(2S)-2-amino-3-(7-methyl-1H-indol-3-yl)propionic acid (2.5 equivalents, 2.5 ml, 100 mM, in DMF) was added to the resin, followed by the addition of HATU (5 equivalents, 1 ml, 500 mM, in DMF) and DIPEA (13 equivalents, 0.65 ml, 2 M, in NMP). The mixture was stirred by bubbling with nitrogen for 40 min and then purged. The resin was washed once with 4 ml DMF, and the coupling process was repeated once. Before starting the next deprotection-coupling cycle, the resin was washed five times with 4 ml DMF by bubbling with N2. Step XI: Coupling of Fmoc-11-aminoundecanoic acid (Aud):

[0718] To deprotect the resin-bound Fmoc groups, add 20% piperidine from 4 ml of DMF to the swollen resin and mix by bubbling with nitrogen for 2.5 min, then purge. Repeat this process once.

[0719] After deprotection, the resin was washed 7 times with 4 ml DMF under mixed conditions. A solution of Fmoc-11-aminoundecanoic acid (2.5 equivalents, 2.5 ml, 100 mM, in DMF) was added to the resin, followed by HATU (5 equivalents, 1 ml, 500 mM, in DMF) and DIPEA (13 equivalents, 0.65 ml, 2 M, in NMP). The mixture was stirred by bubbling with nitrogen for 40 min, then purged. The resin was washed once with 4 ml DMF, and the coupling process was repeated once. Before starting the next deprotection-coupling cycle, the resin was washed 5 times with 4 ml DMF by bubbling with N2. Resin Cutting

[0720] The constructed peptide was isolated from the resin and protecting groups by cleavage with Kings mixture followed by precipitation with diethyl ether. cyclization

[0721] The crude peptide was dissolved in DMF (10 mg / ml), and then PyBOP (1.2 equivalents) and DIPEA (4 equivalents) were added. The mixture was stirred at room temperature for 2 hours until lactamation was complete. Purification was then performed by RP-HPLC.

[0722] Freeze-drying of the pure fraction yielded the desired product peptide SEQ ID NO: 4. Example 2: Peptide Cyclation Cyclization via amide bond formation

[0723] The crude peptide was dissolved in DMF (10 mg / ml), and then PyBOP (1.2 equivalents) and DIPEA (4 equivalents) were added. The mixture was stirred at room temperature for 2 hours until lactamation was complete. Purification was performed by RP-HPLC and lyophilization of the pure fraction.

[0724] Alternatively, the peptide was dissolved in DMF, cooled to 0°C, and then 20 equivalents of NaHCO3, 3 equivalents of HOAT, and 2.5 equivalents of EDC.HCl were added. The reaction was stirred and allowed to slowly equilibrate to room temperature overnight. The NaHCO3 was filtered under gravity, and the soluble fraction was dried by rotary evaporation. The crude cyclic peptide was dissolved in ACN:water and freeze-dried. The desired cyclic peptide product was isolated by a preparative RP-HPLC purification procedure.

[0725] In the case of Yae in the sequence, after cleaving the mixture from the resin using King cleavage, the N-terminus retains Fmoc protection, and the deprotected Yae moiety is protected with BOC-OSu (2 equivalents) and diisopropyl-ethylamine in DMF. Then, the N-terminal Fmoc group is cleaved with piperidine (20 equivalents).

[0726] Following post-processing and RP-HPLC purification, peptide cyclization was performed. Finally, the BOC protecting group was cleaved using TFA (50 equivalents). The desired cyclic peptide product was then isolated using a standard preparative RP-HPLC purification procedure. Cyclization via alkylation with succinaldehyde

[0727] After cleavage from the resin and purification by HPLC, the freeze-dried peptide was dissolved in methanol. Sodium cyanoborohydride (1 equivalent), succinaldehyde (1 equivalent), and DIPEA (1 equivalent) were added, and the mixture was stirred for 16 hours. The solvent was evaporated, and the residue was purified by HPLC and freeze-dried to obtain the desired peptide. Coupling of PyEA with the C-terminus

[0728] DIPEA (0.25 mmol) was added to a solution of a peptide (0.08 mmol) with a carboxylic acid functional group at the C-terminus in DMF (20 mL), and the resulting mixture was stirred at 0°C for 0.5 h. Then, DEPBT (0.088 mmol) and PyEA (0.08 mmol) were added sequentially. The mixture was stirred at 0°C for 1 h and then at room temperature for 1 h. The mixture was concentrated under vacuum, diluted with water (500 mL), and lyophilized. The crude product was purified by chromatography.

[0729] The peptide compounds obtained by this method are shown in Table 3: Table 3: Synthesized peptide compounds Example 3: Analytical HPLC / UHPLC

[0730] Method A: Detection at 214 nm

[0731] HPLC: Waters Class i UPLC

[0732] Column: Waters ACQUITY UPLC® CSH™ C18 1.7 µm (75 × 2.1 mm), 50°C; Solvent: H2O + 0.05% TFA: ACN + 0.045% TFA (Flow rate: 0.7 ml / min)

[0733] Gradient: 95:5 (0 min) to 95:5 (1 min) to 20:80 (12 min) to 5:95 (12.5 min) to 5:95 (13.5 min) to 95:5 (14 min) to 95:5 (16 min)

[0734] Features a mass analyzer: Waters Xevo G2-XS QTof, electrospray cation mode, mass range: 300-3200 m / z, resolution mode.

[0735] Method B: Detection at 214 nm

[0736] HPLC: Agilent 1290 Infinity II UPLC

[0737] Column: Waters ACQUITY UPLC® CSH™ C18 1.7 µm (150 × 2.1 mm), 50°C; Solvent: H2O + 0.05% TFA: ACN + 0.035% TFA (Flow rate: 0.5 ml / min)

[0738] Gradient: 80:20 (0 min) to 80:20 (3 min) to 25:75 (23 min) to 2:98 (23.5 min) to 2:98 (30.5 min) to 80:20 (31 min) to 80:20 (37 min)

[0739] Features a mass analyzer: Agilent 6230 Precision Mass TOF, Agilent Dual Jet ESI+, mass range: 300-3200 m / z, extended dynamic range mode (2 GHz).

[0740] Method C: Detection at 214 nm

[0741] HPLC: Agilent 1290 Infinity II UPLC

[0742] Column: Waters ACQUITY UPLC® CSH™ C18 1.7 µm (150 × 2.1 mm), 50°C; Solvent: H2O + 0.05% TFA: ACN + 0.045% TFA (Flow rate: 0.5 ml / min)

[0743] Gradient: 95:5 (0 min) to 95:5 (3 min) to 20:80 (30 min) to 2:98 (30.5 min) to 2:98 (34.5 min) to 95:5 (35 min) to 95:5 (40 min)

[0744] Features a mass analyzer: Agilent 6230 Precision Mass TOF, Agilent Dual Jet ESI, mass range: 300-3200 m / z, extended dynamic range mode (2 GHz). Table 4: List of synthesized peptides, comparison of calculated and measured molecular weights, and HPLC / UHPLC methods used. Example 4: Typical Preparative HPLC Purification Procedure

[0745] The crude peptide was purified using an Äkta purification system, a Jasco semiprep HPLC system, a Waters automated purification system, an Agilent 1100 HPLC system, or a similar HPLC system. Depending on the amount of crude peptide to be purified, preparative RP-C18-HPLC columns of different sizes and flow rates were used, for example: Waters XSelect CSH C18 OBDPrep 5 μm 30 × 250 mm, Waters SunFire C18 OBD Prep 5 μm 30 × 250 mm, Waters SunFire C18 OBD Prep 5 μm 50 × 150 mm, and Phenomenex Luna Prep C18 5 µm 21.2 × 250 mm. Acetonitrile (B) and water + 0.1% TFA (A) or water + 0.1% FA (A) were used as eluents. The fraction containing the product was collected and lyophilized to obtain the purified product, typically a TFA salt.

[0746] Alternatively, the peptide can be isolated as acetate using the following procedure: Dissolve the peptide in water and adjust the solution to pH 7.05 with NaHCO3. Then, purify the dissolved compound using an RP Kinetex 21, 2 × 250 mm column (column volume CV 88 ml, 5 µm, C18, 100A, Äkta avant 25): Equilibrate the column with solvent A (3 × CV), inject the compound, and then wash with a mixture of solvent A (95%) and solvent B (5%) at 3 CV. Then, run a gradient solvent A:B (95:5) to A:B (20:80) at 15 CV. Collect the purified peptide and lyophilize it. Example 5: Solubility test of the example at different pH values

[0747] Before measuring the solubility of peptide batches, their purity was determined by UHPLC / MS method A.

[0748] For solubility testing, the target concentration was 1 mg of pure compound / mL. Therefore, a solution of the solid sample was prepared in a buffer system with a compound concentration of 1 mg / mL. Solid samples were obtained by drying aliquots of the freshly prepared dimethyl sulfoxide stock solution using a THERMO SPEEDVAC vacuum centrifuge for 3 hours.

[0749] The following buffering system should be used:

[0750] Solubility buffer system A) 50 mM HCl pH 1.2

[0751] Solubility buffer system B) 50 mM phosphate buffer, pH 6.5

[0752] Solubility buffer system C) 50 mM phosphate buffer, pH 7.4

[0753] After gently stirring the filtered solution (0, 45 µm filter plate) for 2 hours, UHPLC-UV was performed using analytical method A. The filtered solution was obtained by forced filtration by centrifugation at 1500 RCF (relative centrifugal acceleration) for 10 min.

[0754] Solubility was determined by comparing the UV peak area of ​​a 2 µL injection of a 1:10 diluted buffer sample with a standard curve of a dimethyl sulfoxide reference sample of known concentration. Different extinction coefficients were considered in the calculations.

[0755] The results are shown in Table 5. Table 5: Solubility of peptide compounds in buffer formulations Example 6: Chemical stability assessment of example compounds

[0756] The purity of the peptide batch was determined by UHPLC / MS method A prior to chemical stability measurements.

[0757] For stability assessment, the target concentration was 0.5 mg of pure compound / mL. Therefore, a solution of the solid sample was prepared in a buffer system with a compound concentration of 0.5 mg / mL. Solid samples were obtained by drying aliquots of the freshly prepared acetonitrile stock solution using a THERMO SPEEDVAC vacuum centrifuge for 3 hours.

[0758] The following buffering system should be used:

[0759] Solubility buffer system A): 50 mM HCl pH 1.2 + acetonitrile (3:1).

[0760] Solubility buffer system B): 50 mM phosphate buffer pH 6.5 + acetonitrile (3:1).

[0761] Solubility buffer system C): 50 mM phosphate buffer pH 7.4 + acetonitrile (3:1).

[0762] The prepared solutions were stored at 2°C and 37°C for 24 hours.

[0763] After this time interval, the sample was centrifuged at 2500 RCF for 15 min to produce a 1:5 diluted supernatant. Then, 2 µl of the diluted solution was analyzed by UHPLC-UV using Method A.

[0764] Chemical stability is assessed by calculating the relative purity loss using the following formula:

[0765] [(Purity after 0 hours at 37°C) - (Purity after 24 hours at 37°C)] / (Purity after 0 hours at 37°C)] 100%

[0766] Purity is calculated as follows: [(peak area of ​​peptide) / (total peak area)] 100%.

[0767] The results are shown in Table 6. Table 6: Chemical stability (relative purity loss) of peptide compounds in buffer formulations Example 7: Evaluation of peptide protease stability

[0768] Studies were conducted in simulated intestinal fluid (SIF) and simulated gastric fluid (SGF) to evaluate the gastric stability of the peptide compounds disclosed herein.

[0769] For clarification, the term "psepsin-SGF" refers to a solution of simulated gastric juice containing pepsin, while "SGF medium" refers to a solution of simulated gastric juice without pepsin. Pepsin-SGF is prepared by dissolving 320 mg of pepsin (Merck Millipore) in 100 mL of SGF medium concentrate (Prosense).

[0770] Similarly, the term "pancreatic secretase-SIF" refers to a solution of simulated intestinal fluid containing pancreatic secretase, while "SIF medium" refers to a solution of simulated intestinal fluid without pancreatic secretase. Pancreatic secretase-SIF is prepared by dissolving 1 mg of pancreatic secretase (MP Biomedicals) in 10 mL of SIF medium concentrate (Purson).

[0771] The peptide compound was first dissolved in 0.01 HCl (+ / - 10% DMSO) at a stock concentration of 1 mM, and then further diluted with water to a stock concentration of 20 µM. The final peptide concentration during incubation was 1 µM.

[0772] Add 4 µL of 20 µM peptide stock solution to 76 µL of pepsin-SGF buffer and incubate at 37°C. At each time point (0, 60, 120, and 240 min), quench the reaction by adding 80 µL of ethanol (0.1% HCOOH). Centrifuge the sample for 20 min, and transfer aliquots of 100 µL of supernatant to LoBind-MTP plates for analysis by LCMS / MS. Calculate the peak area response for each test compound at each time point. As an internal reference, calculate the peak area response of the same test compound at T0. Calculate the percentage remaining at each time point based on the peak area response ratio of the test compound at T0. Set time 0 to 100%, and calculate all subsequent time points relative to time 0.

[0773] The peptide compound was first dissolved in 0.01 HCl (+ / - 10% DMSO) at a stock concentration of 1 mM, and then further diluted with water to a stock concentration of 20 µM. The final peptide concentration during incubation was 1 µM.

[0774] Add 4 µL of 20 µM peptide stock solution to 76 µL of pepsin-SGF buffer and incubate at 37°C. At each time point (0, 60, 120, and 240 min), quench the reaction by adding 80 µL of ethanol (0.1% HCOOH). Centrifuge the sample for 20 min, and transfer aliquots of 100 µL of supernatant to LoBind-MTP plates for analysis by LCMS / MS. Calculate the peak area response for each test compound at each time point. As an internal reference, calculate the peak area response of the same test compound at T0. Calculate the percentage remaining at each time point based on the peak area response ratio of the test compound at T0. Set time 0 to 100%, and calculate all subsequent time points relative to time 0. Table 7: Protease stability in simulated gastric juice (FaSSGF) and simulated intestinal juice (FaSSIF), expressed as the remaining percentage of peptide compounds.

[0775] na: Not available. Example 8: Assay for a specific human IL-23R inhibitor

[0776] The antagonistic effect of the compound on human interleukin-23 receptor inhibitors was determined by ELISA assay.

[0777] A 384-well plate was coated with 50 µl / well of human IL23 at a final concentration of 1 µg / mL and incubated overnight at 4°C. The wells were washed three times with 80 µl of wash buffer and blocked for 60 min at room temperature with 80 µl of blocking buffer, followed by another wash. 20 µl of serially diluted test peptide was added to each well, followed by 20 μl of recombinant human IL-23R-Fc chimera, to a final concentration of 0.1 μg / mL. The plate was incubated at room temperature for 60 min. After washing the wells, the bound IL23R-Fc was detected using goat anti-human IgG1-HRP antibody. The signal was visualized using QUANTABLU fluorescent peroxidase substrate.

[0778] To improve the selectivity of IL23R inhibitor peptides in binding to human IL12Rβ1, recombinant human IL-12Rβ1-Fc was used for ELISA assay.

[0779] To determine the selectivity of IL23R inhibitor peptides in binding to mouse IL23R, recombinant mouse IL-23R-Fc chimeras were used, and the binding of mouse IL23R-FC was detected using goat anti-mouse IgG1-HRP antibody.

[0780] The results are presented in Table 8 below. Table 8: IC50 values ​​for inhibiting the binding of IL-23 to IL-23R, as measured by ELISA 50 value Example 9: Inhibition of IL-23-induced STAT3 signaling in HEKBlue™ IL-23 cells

[0781] HEK-Blue™ IL-23 cells (INVIVOGEN) are designed to detect bioactive human (hIL-23) and mouse IL-23 (mIL-23) by monitoring activation of the STAT3 pathway. The binding of IL-23 to its receptor on the surface of HEK-Blue™ IL-23 cells triggers a signaling cascade, leading to STAT3 activation and subsequent production of secreted embryonic alkaline phosphatase (SEAP). This can be easily assessed using QUANTI-Blue™ solutions (SEAP assay kit).

[0782] Culture HEK-Blue™ IL-23 cells as described by Ingenium.

[0783] On day 1 of the assay, 10 µL of HEK-BLUE IL23 cell suspension (final concentration: 10,000 cells / well) and 10 µL of serially diluted peptide solution were dispensed into 384-well assay plates. After pre-incubating the reaction mixture at 37°C / 5% CO2 / 95% humidity for 60 min, 10 µL of human IL23 cytokine (final concentration: 0.25 ng / ml) was added to each well for STAT3 signaling activation. The plates were incubated overnight at 37°C / 5% CO2 / 95% humidity.

[0784] On the second day, 20 µl / well of QUANTI-BLUE assay solution (prepared according to manufacturer's instructions) was dispensed into new 384-well plates, followed by a transfer of 5 μl of the supernatant from the overnight incubated cell / sample mixture. After incubation at room temperature for 45 min, the signal at OD 620–655 nm was measured using a Pherastrar (BMG).

[0785] The dose-response data were fitted using XLfit. Table 9: IC50 of IL-23-induced pSTAT3 inhibition as measured by SEAP reporter genes phosphorylated by STAT3 50 value [References] Atherton E. and R. C. Sheppard, Solid Phase Peptide Synthesis. APractical Approach, Oxford-IRL Press, New York, 1989 Bis RL, Mallela KM. Antimicrobial preservatives induce aggregation ofinterferon alpha-2a: the order in which preservatives induce proteinaggregation is independent of the protein. Int J Pharm. 2014 Sep 10;472(1-2):356-61. doi: 10.1016 / j.ijpharm.2014.06.044. Epub 2014 Jun 27. PMID: 24974985;PMCID: PMC4268133. Brayden DJ, Mrsny RJ. Oral peptide delivery: prioritizing the leadingtechnologies. Ther Deliv. 2011 Dec;2(12):1567-73. doi: 10.4155 / tde.11.114.PMID: 22833982. David B. Troy, Paul Beringer. Remington: The Science and Practice ofPharmacy Lippincott Williams & Wilkins, 2006 - 2393 pages Kamerzell TJ, Esfandiary R, Joshi SB, Middaugh CR, Volkin DB.Protein-excipient interactions: mechanisms and biophysical characterizationapplied to protein formulation development. Adv Drug Deliv Rev. 2011 Oct;63(13):1118-59. doi: 10.1016 / j.addr.2011.07.006. Epub 2011 Jul 29. PMID:21855584. King DS, Fields CG, Fields GB. A cleavage method which minimizes sidereactions following Fmoc solid phase peptide synthesis. Int J Pept ProteinRes. 1990 Sep;36(3):255-66. doi: 10.1111 / j.1399-3011.1990.tb00976.x. PMID:2279849. Rowe, Raymond C. et al. “Handbook of Pharmaceutical Excipients.”(1994). Siri Ram Chhabra, Bhupinder Hothi, David J. Evans, Peter D. White,Barrie W. Bycroft, Weng C. Chan, An appraisal of new variants of Dde amineprotecting group for solid phase peptide synthesis, Tetrahedron Letters,Volume 39, Issue 12, 1998, Pages 1603-1606, ISSN 0040-4039, https: / / doi.org / 10.1016 / S0040-4039(97)10828-0. Sayago C, Gonzalez Valcarcel IC, Qian Y, Lee J, Alsina-Fernandez J,Fite NC, Carrillo JJ, Zhang FF, Chalmers MJ, Dodge JA, Broughton H, Espada A.Deciphering Binding Interactions of IL-23R with HDX-MS: Mapping Protein andMacrocyclic Dodecapeptide Ligands. ACS Med Chem Lett. 2018 Aug 1;9(9):912-916. doi: 10.1021 / acsmedchemlett.8b00255. PMID: 30258540; PMCID: PMC6142055. Stewart John Morrow and Young Janis Dillaha. Solid phase peptidesynthesis. Pierce Chemical Co., Rockford, Ill. et ©1984 Theodora W. Greene Ph.D.,, Peter G. M. Wuts Ph.D., Protective Groupsin Organic Synthesis, Third Edition. Copyright © 1999 by John Wiley & Sons,Inc. Print ISBN:9780471160199 |Online ISBN:9780471220572 |DOI:10.1002 / 0471220574 Tuvia S, Pelled D, Marom K, Salama P, Levin-Arama M, Karmeli I,Idelson GH, Landau I, Mamluk R. A novel suspension formulation enhancesintestinal absorption of macromolecules via transient and reversibletransport mechanisms. Pharm Res. 2014 Aug;31(8):2010-21. doi: 10.1007 / s11095-014-1303-9. Epub 2014 Feb 21. PMID: 24558008; PMCID: PMC4153969. US 2013-029907 A1 WO 2016 / 011208 A1 WO 2017 / 011820 A2 WO 2018 / 089693 A2 WO 2018 / 022937 A1 WO 2018 / 136646 A1 WO 2022 / 109328 A1 WO 2013 / 063468 A1 WO 2015 / 179438 A1。

Claims

1. A peptide compound of formula (I): X1(§1)(#1)-X2(#1)(@2)-X3-X4-X5-X6-X7-X8-X9(§1)(@2)-X10-X11-X12-X13-R1(I), in X1 is absent or has a butyl portion and When X1 is a butyl group, then (§1) represents the alkyl bond between X1 and X9, and (#1) represents the alkyl bond between X1 and X2, or When X1 is absent, (@2) represents the amide bond between X2 and X9. X2 is an amino acid residue selected from the following: Add, Ade, Ahd, Aoa, Atd, Atea, Aud, Dad, Dae, Dec, Dod, Dpa, Kme, Kme(αAc), Lys(αAc), Nmapt, Nmha, and Oca. X3 is an amino acid residue selected from the following: Nak, Trp, Wdm, Wim, Wme, Wcl, and Wfl; X4 is an amino acid residue selected from the following: Aib, Gln, Glu, Ile, Iva, Leu, Lys, Lys(Ac), Mle, Mly, Mva, Phe, Thr, Trp, and Val; X5 is an amino acid residue selected from the following: Aib, Mkdm, Mle, and Thp; X6 is an amino acid residue selected from the following: Trp, Tyr, Yae, Yde, and Yme; X7 is an amino acid residue selected from the following: Nak, Nal, and Trp; X8 is an amino acid residue selected from the following: Aib, Cba, Cit, Gln, Leu, Lys, Lys(Ac), Mle, Mly, D-Mkdm, Mkdm, Thp, D-Trp, and D-Tza; X9 is an amino acid residue selected from the following: Asp, Glu, Kme, Lys, and Orn; X10 is an amino acid residue selected from the following: Ala, Asn, Gly, and Ser; X11 is an amino acid residue selected from the following: Ala, Asn, Bal, Gly, Hol, Hph, His, Ile, Iva, Leu, Mhis, Pal, Pyal, PyEA, Val, and their corresponding D-forms; X12 is absent or consists of amino acid residues selected from the following: Bal, Dnmy, Ile, Lys, Mep, Mys, and Sar; X13 is absent or is a Lys residue; and... R1 either does not exist or is selected from -NH2, -OH and -N(C2H5)2; Or its salts or solvates.

2. The peptide compound according to claim 1, wherein... X1 is the butyl moiety, and X9 is a residue of Kme, or X1 is absent, X2 is an amino acid residue selected from the following: Dad, Dae, Dec, Dod, Dpa, and Oca, and X9 is an amino acid residue selected from the residues Kme, Lys, and Orn, or X1 is absent, X2 is an amino acid residue selected from the following: Dec, Dod, Dpa, and Oca, and X9 is an amino acid residue selected from the following: Kme and Lys, or X1 is absent, X2 is an amino acid residue selected from the following: Add, Ade, Ahd, Aoa, Atd, Atea, Aud, Kme, Kme(αAc), Lys(αAc), Nmapt, and Nmha, and X9 is an amino acid residue selected from the following: Asp and Glu, or X1 does not exist, X2 is an amino acid residue selected from the following: Add, Ade, Ahd, Aoa, Atd, Atea, and Aud, and X9 is a residue of Glu.

3. The peptide compound according to claim 1, wherein... X2 is an amino acid residue selected from the following: Add, Ade, Ahd, Aoa, Atd, Atea, Aud, Dec, Dod, Dpa, Kme, Kme(αAc), Nmapt, Nmha, and Oca; X3 is an amino acid residue selected from the following: Trp, Wdm, Wim, and Wme; X4 is an amino acid residue selected from the following: Gln, Leu, and Lys(Ac); X5 is an amino acid residue selected from the following: Aib and Thp; X6 is a residue of Yde; X7 is a residue of Nal; X8 is an amino acid residue selected from the following: Aib, Mkdm, and Thp; X9 is an amino acid residue selected from the following: Glu, Kme, and Lys; X10 is an amino acid residue selected from the following: Asn, Gly, and Ser; X11 is an amino acid residue selected from the following: Pal and PyEA; X12 is absent or is a residue of Sar; X13 does not exist; and... R1 is either absent or -NH2.

4. The peptide compound according to claim 3, wherein... X10 represents Asn; or, X10 represents an amino acid residue selected from the following: Gly and Ser, and X12 represents Sar; and R1 represents NH2, or R1 represents NH2 and When X1 is a butyl moiety, X2 represents an amino acid residue selected from the following: Kme, Nmapt, and Nmha; or When X1 is absent, X2 represents an amino acid residue selected from Add, Ade, Aoa, Atd, Atea, Aud, Dec, Dod, and Oca.

5. The peptide compound according to claim 1 or 2, wherein... X2 is an amino acid residue selected from the following: Add, Ade, Ahd, Aoa, Atd, Atea, Aud, Dec, Dod, Dpa, Kme, Kme(αAc), Nmapt, Nmha, and Oca; X3 is an amino acid residue selected from the following: Trp, Wim, and Wme; X4 is an amino acid residue selected from the following: Gln, Leu, and Lys(Ac); X5 is an amino acid residue selected from the following: Aib and Thp; X6 is a residue of Yde; X7 is a residue of Nal; X8 is an amino acid residue selected from the following: Aib, Mkdm, and Thp; X9 is an amino acid residue selected from the following: Glu, Kme, and Lys; X10 is an amino acid residue selected from the following: Asn, Gly, and Ser; X11 is an amino acid residue selected from the following: Pal and PyEA; X12 is absent or is a residue of Sar; X13 does not exist; R1 is absent or -NH2. in When X2 is a residue of Nmapt, X11 is a residue of PyEA, and X4 is a residue of Gln, then X10 is not a residue of Gly, or When X2 is a residue of Nmapt, X11 is a residue of PyEA, and X10 is a residue of Gly, then X4 is not a residue of Gln, or When X2 is a residue of Nmha and X11 is a residue of PyEA, then X4 is not a residue of Lys(Ac).

6. The peptide compound according to claim 1 or 3, wherein... X2 is an amino acid residue selected from the following: Add, Ade, Ahd, Aoa, Atd, Atea, Aud, Dec, Dod, Dpa, Kme, Kme(αAc), Nmapt, Nmha, and Oca; X3 is an amino acid residue selected from the following: Trp, Wim, and Wme; X4 is an amino acid residue selected from the following: Gln, Leu, and Lys(Ac); X5 is an amino acid residue selected from the following: Aib and Thp; X6 is a residue of Yde; X7 is a residue of Nal; X8 is an amino acid residue selected from the following: Aib, Mkdm, and Thp; X9 is an amino acid residue selected from the following: Glu, Kme, and Lys; X10 is an amino acid residue selected from the following: Asn, Gly, and Ser; X11 is an amino acid residue selected from the following: Pal and PyEA; X12 is absent or is a residue of Sar; X13 does not exist; and R1 is absent or -NH2; in When X2 is a residue of Nmapt, X11 is a residue of PyEA, and X4 is a residue of Gln, then X10 is not a residue of Gly, or When X2 is a residue of Nmapt, X11 is a residue of PyEA, and X4 is a residue of Lys(Ac), then X10 is not a residue of Ser, or When X2 is a residue of Nmapt, X11 is a residue of PyEA, and X10 is a residue of Gly, then X4 is not a residue of Gln, or When X2 is a residue of Nmha, then X11 is not a residue of PyEA.

7. The peptide compound according to claim 1 or 3, wherein... X2 is an amino acid residue selected from the following: Add, Ade, Ahd, Aoa, Atd, Atea, Aud, Dec, Dod, Kme, Kme(αAc), Nmapt, Nmha, and Oca; X3 is an amino acid residue selected from the following: Trp, Wim, and Wme; X4 is an amino acid residue selected from the following: Gln, Leu, and Lys(Ac); X5 is an amino acid residue selected from the following: Aib and Thp; X6 is a residue of Yde; X7 is a residue of Nal; X8 is an amino acid residue selected from the following: Aib, Mkdm, and Thp; X9 is an amino acid residue selected from the following: Glu, Kme, and Lys; X10 is an amino acid residue selected from the following: Asn, Gly, and Ser; X11 is an amino acid residue selected from the following: Pal and PyEA; X12 is absent or is a residue of Sar; X13 does not exist; and R1 is absent or -NH2; in When X2 is a residue of Nmapt, then X11 is not PyEA, or When X2 is a residue of Nmapt and X11 is a residue of PyEA, then X3 is a residue of Wim, X4 is a residue of Gln, and X10 is a residue of Ser, or When X2 is a residue of Nmha, then X11 is not a residue of PyEA, or If X2 is a residue of Aoa or Dec, then X4 is not a residue of Leu.

8. The peptide compound according to claim 1 or 3, wherein X2 is an amino acid residue selected from the following: Add, Ade, Aoa, Atd, Atea, Aud, Dec, Dod, Kme, Kme(αAc), Nmapt, Nmha, and Oca; X3 is an amino acid residue selected from the following: Trp, Wim, and Wme; X4 is an amino acid residue selected from the following: Gln, Leu, and Lys(Ac); X5 is an amino acid residue selected from the following: Aib and Thp; X6 is a residue of Yde; X7 is a residue of Nal; X8 is an amino acid residue selected from the following: Aib, Mkdm, and Thp; X9 is an amino acid residue selected from the following: Glu, Kme, and Lys; X10 is an amino acid residue selected from the following: Asn, Gly, and Ser; X11 is a residue of Pal; X12 is a residue of Sar; X13 does not exist; and R1 is -NH2; in, When X2 is a residue of Nmapt and X3 is a residue of Wme, then X8 is not a residue of Mkdm, or When X2 is a residue of Aoa, then X4 is not a residue of Leu.

9. The peptide compound according to any one of claims 1 to 8, wherein the peptide compound is represented by SEQ ID NO: 1 to 50, or a salt or solvation thereof.

10. The peptide compound of claim 9, wherein the peptide compound is represented by a sequence selected from: -SEQ ID NO: 1-22, 24, 26-48 and 50, or their salts or solvates; or -SEQ ID NO: 1-21, 24, 26-36, 38-48 and 50, or their salts or solvates; or -SEQ ID NO: 1-8, 10-14, 16, 18-21, 24, 26-36, 39-47 and 50, or their salts or solvates; or -SEQ ID NO: 1-2, 4-8, 10-14, 16, 18-21, 24, 26-35, 39-44, 46-47 and 50, or their salts or solvates; or -SEQ ID NO: 1-2, 4-5, 7-8, 10-14, 16, 18-21, 26, 28-29, 31-35, 39-40, 43-44, 46-47 and 50, or their salts or solvates; or -SEQ ID NO: 1-2, 4-5, 7-8, 10-14, 16, 18-21, 31-35, 39, 44, 46-47 and 50, or their salts or solvates; or -SEQ ID NO: 4, 11, 16, 18, 19 and 50, or their salts or solvates.

11. The peptide compound according to any one of claims 1 to 10, wherein the peptide compound is in a concentration of about 50 nM or less, or about 25 nM or less, or about 15 nM or less, or about 10 nM or less, or about 5 nM or less, or about 3 nM or less, or about 2 nM or less, or about 1 nM or less. 50 Inhibit IL-23-induced STAT3 phosphorylation.

12. The peptide compound according to any one of claims 1 to 11, wherein the peptide compound has at least 50% protease stability, which is measured as the percentage of the remaining portion of the peptide compound in simulated intestinal fluid at 37°C for 60 min.

13. The peptide compound according to any one of claims 1 to 12, for use in treating and / or preventing autoimmune or inflammatory diseases in a subject of need.

14. The peptide compound for use according to claim 13, wherein the autoimmune or inflammatory disease is selected from inflammatory bowel diseases such as Crohn's disease and ulcerative colitis, psoriasis, psoriatic arthritis, and hidradenitis suppurativa.

15. A pharmaceutical composition comprising at least one peptide compound according to any one of claims 1 to 12, or a pharmaceutically acceptable salt or solvate thereof, and at least one pharmaceutically acceptable excipient.

16. A method for treating and / or preventing autoimmune or inflammatory diseases, the method comprising at least the step of administering a peptide compound according to any one of claims 1 to 12 to the subject.

17. Use of the peptide compound according to any one of claims 1 to 12 in the preparation of a medicament for the treatment and / or prevention of autoimmune or inflammatory diseases in a subject of need.

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