IL-17A binding peptide and its medical use

Abstract

The present invention relates to IL-17A binding peptides, inhibitors of the interaction between IL-17A and receptor IL-17RA, and bioconjugates, dimers, pharmaceutical compositions and their medical uses.

Description

[0001] The present invention relates to IL-17A binding peptides, inhibitors of IL-17A binding to IL-17AR receptors to form IL-17A / IL-17RA / IL17RC complexes, bioconjugates and dimers containing the above peptides, pharmaceutical compositions and medical uses of the above peptides, bioconjugates and dimers. Background of the Invention

[0003] Human cytotoxic T lymphocyte-associated antigen 8 (CTLA8), also known as interleukin 17 (IL-17), was first cloned in 1993. The first biological activity described for human IL-17 was to induce rheumatoid arthritis synoviocytes to produce interleukin 6 (IL-6) and interleukin 8 (IL-8). This finding suggests that IL-17 plays a role in inflammation through IL-6 and in neutrophil recruitment through IL-8 (Xu S et al, Cell Mol Immunol. 2010, 164-74; Murcia RY et al, PLOS ONE 2016, 11 (5): e0154755 doi: 10.1371 / journal.pone.0154755).

[0004] It was later discovered that this molecule (later called IL-17A) is part of a larger family that includes five other members, IL-17B-F. IL-17F shares the greatest homology (about 56%) with the first IL-17 discovered, IL-17A, while IL-17E exhibits the lowest sequence conservation (about 16%). Members of the IL-17 family function as homodimers to bind to their receptors, with the exception of IL-17A and IL-17F, which can also act as heterodimers (Goepfert A et al, Sci Rep. 2017, 7(1), 8906).

[0005] IL-17A, the most extensively studied cytokine in this family, plays a crucial role in host defense against microbial infection and is considered a major driver of many inflammatory and autoimmune conditions. Pathological production of IL-17A leads to excessive inflammation and significant tissue damage.

[0006] In particular, IL-17A can amplify a series of events leading to neutrophil recruitment, inflammation, and host defense by inducing a variety of molecules including cytokines, chemokines, acute phase proteins, antimicrobial peptides, mucins, and matrix metalloproteinases. Although IL-17A is a signature cytokine produced by T helper 17 (Th17) cells, IL-17A, like other IL-17 family cytokines, has multiple sources ranging from immune cells to non-immune cells.

[0007] IL-17B, IL-17C, and IL-17D are also considered pro-inflammatory cytokines, but their roles are not fully understood. IL-17E, also known as IL-25, has the lowest homology and is involved in Th2 cell responses to parasites and allergic reactions. CCL20 drives the recruitment of Th17 cells and dendritic cells to sites of inflammation. In turn, Th17 cells are activated, producing inflammatory mediators and leading to chronic inflammation.

[0008] Cytokines belonging to the IL-17 family signal through their cognate receptors and activate downstream pathways including NFκB, MAPK, and C / EBP, thereby inducing the expression of antimicrobial peptides, cytokines, and chemokines. The proximal adaptor Act1 is a common mediator in all IL-17 cytokine signaling processes and is involved in IL-17-mediated host defense as well as IL-17-driven autoimmune conditions.

[0009] The IL-17 receptor family consists of five members, namely IL-17RA, IL-17RB, IL-17RC, IL-17RD and IL-17RE, all of which share sequence homology with their ligands.

[0010] IL-17RA is ubiquitously expressed in a wide range of tissues and cell types and binds to IL-17-A, IL-17-C, IL-17-E, and IL-17-F. Receptor signaling occurs through a heterodimeric receptor formed by a common IL17-RA subunit and a second subunit that is ligand-dependent and modulates signaling specificity. Upon stimulation with a ligand, IL-17RA forms a heterodimeric receptor complex with IL-17RB (for IL-17E), IL-17RC (for IL-17A and IL-17F), or IL-17RE (for IL-17C). It is proposed that the binding of a ligand to the first IL-17RA receptor subunit promotes a second binding event, thereby inducing the formation of a heterodimeric receptor complex. In particular, the signals of IL-17A and IL-17F are mediated by a complex between IL-17RA and IL-17RC. IL-17F binds to IL-17RA with an affinity approximately 100 to 1000 times lower than that of IL-17A, while the binding affinity of the two cytokines to IL-17RC is comparable (Onishi RM et al, Immunology 2010, 129(3), 311-321; Gu C et al, Cytokine 2013, 64(2), 477-485).

[0011] Human IL-17A plays a key role in various autoimmune and inflammatory conditions, such as rheumatoid arthritis, multiple sclerosis, psoriasis, Crohn's disease, systemic lupus erythematosus, asthma, Behcet's disease and hyper-IgE syndrome (Fujino S et al Gut, 2003, 52, 65-70; Shabgah AG et al, Postepy Dermatol Allergol 2014, 31 (4), 256-261). In addition, IL-17A blockade has shown preclinical and clinical efficacy in ankylosing spondylitis and rheumatoid arthritis (Liu S et al, Nature-Scientific Reports 2016, doi: 10.1038 / srep26071; Lubberts E et al, Arthritis Rheum 2004, 50, 650-659).

[0012] Moreover, IL-17A has been shown to be associated with ocular diseases, particularly with the pathogenesis of ocular surface and corneal diseases such as dry eye disease (DED), viral and bacterial keratitis. Antibodies developed to neutralize IL-17A have shown promising results in reducing the severity of these diseases (Garbutcheon-Singh KB et al, Curr Eye Res, 2018, DOI: 10.1080 / 02713683.2018.1519834).

[0013] DED is an inflammatory and autoimmune ophthalmic disease of the tear system and ocular surface that causes discomfort, visual impairment, and tear film damage. DED is one of the most common causes of acquired visual impairment in the adult population (Stern ME et al, Mucosal Immunol 2010, 3 (5), 425-442; Stevenson W et al, Arch Ophthalmol 2012, 130 (1), 90-100, Parul Singh, Parul Singh 2012, Hah, Chung et al. 2017). The disease involves immune and inflammatory processes that affect the ocular surface and may cause blindness in severe cases. Current treatments for DED are mainly symptomatic, with ocular lubricants and nonspecific anti-inflammatory drugs, such as corticosteroids, cyclosporine A, and tacrolimus.

[0014] IL-17A is associated with impaired corneal epithelial barrier function, which is the most visually threatening complication of DED. It has been reported that Th17 cells are increased in ocular tissues of patients with DED, which induces increased concentrations of IL-6, TGF-β, IL-23, and IL-17A on the ocular surface, as well as increased IL-17 concentrations in tears and increased numbers of Th17 cells on the ocular surface in experimental DED models. Furthermore, it has been demonstrated that neutralization of IL-17A in vivo results in a substantial reduction in disease induction and severity (De Paiva CS et al, Mucosal Immunol. 2009; 2(3): 243–253, Chauhan et al, Mucosal Immunol, 2009, 2(3), 243–253; Chauhan SK et al, Mucosal Immunol 2009, 2(4), 375–376; Chauhan, El Annan et al. 2009, De Paiva, Chotikavanich et al. 2009, de Paiva, Huang et al. 2014, Subbarayal, Chauhan et al. 2016).

[0015] IL-17A is also associated with psoriasis. Psoriasis is a chronic inflammatory disease. It presents as dry, raised, red skin lesions (plaques) covered with silvery scales. There are many clinical phenotypes (i.e., plaques, punctate, pustular, inverted), and the severity of the disease ranges from a few scattered plaques to widespread involvement of the body surface. Individuals with psoriasis have an increased risk of developing other chronic and serious diseases such as psoriatic arthritis, metabolic syndrome, cardiovascular disease, and depression. IL-17A has been shown to play a central role in the pathophysiology of psoriasis (Zeichner JA et al J Clin Aesthet Dermatol 2016, 9(6 Suppl 1), S3–S6). In psoriasis, IL-17 mRNA expression is higher in lesional skin compared to nonlesional skin. In addition, IL-17A levels are significantly correlated with disease severity (Arican O et al. Mediators Inflamm 2005, 2005, 273–279, Takahashi K et al. Clin Exp Dermatol 2010, 35, 645–649). Blockade of IL-17A has been shown to reduce keratinocyte hyperproliferation, T cell infiltration into the dermis, and mRNA expression of key disease-propagating genes (Krueger JG et al, J Allergy Clin Immunol 2012, 130, 145–154).

[0016] The anti-IL-17A antibodies secukinumab (AIN457, ConsentyxTM, fully human IgG1k anti-IL-17A mAb) and ixekizumab (LY2439821, humanized IgG4 antibody) have been tested in clinical trials for psoriasis and are approved for the treatment of moderate to severe psoriasis. During clinical trials, both antibodies were able to reduce the severity of the disease by at least 75% (PASI75) in 80% of the patient population. In addition, current late-stage clinical trials have also shown promising treatment results for ankylosing spondylitis and psoriatic arthritis. Other antibodies are currently in clinical trials for the same indications.

[0017] The above evidence strongly supports the development of molecules that can target IL-17A and inhibit its signal transduction for the treatment of the above pathologies. Until recently, targeting IL17-A and its receptors was still the domain of antibodies. The reason for the preference for antibodies is that cytokines such as IL-17A constitute targets for protein-protein interactions (PPIs) that are difficult to target and have been considered undruggable so far. In fact, the PPI interface is usually flat and lacks the deep sub-pockets and grooves that are usually necessary for binding small molecular weight (SMW) molecules. Nevertheless, in recent years, new studies have identified several SMW IL-17A antagonists that can bind to IL-17A and prevent it from interacting with the IL-17RA receptor, thereby inhibiting IL-17A pathway activation (Espada A et al. J Med Chem 2016, 59 (5), 2255-2260; Ting JP et al PLoS One 2018, 13 (1), e0190850; Liu S et al, Scientific reports, 6: 26071, doi: 10.1038 / srep26071). Among them, a peptide with the sequence IHVTIPADLWDWINK has been described, which is called a high affinity peptide (HAP) (Liu S et al, Scientific reports, 6: 26071, doi: 10.1038 / srep26071).

[0018] However, there remains a need to identify and characterize novel molecules that can bind to IL-17A with high affinity and prevent activation of the IL-17A pathway in order to develop new therapeutic options for treating disorders involving the IL-17A / IL-17RA axis. SUMMARY OF THE INVENTION

[0020] The present inventors unexpectedly discovered a peptide that is capable of binding to a specific site on the N-terminal portion of IL-17A, thereby inhibiting the interaction with IL-17RA and the formation of the IL-17A7IL-17RA / IL17RC complex.

[0021] Inhibition of said interaction blocks the IL-17A signaling pathway.Thus, the peptides according to the invention are able to reduce TH17-related inflammation and the subsequent damage observed in inflammatory and autoimmune diseases in which these cells and IL-17A play a key role.

[0022] As shown in the experimental part, the peptides of the invention possess physicochemical properties that make them particularly suitable for the topical treatment of ophthalmic and dermatological diseases.

[0023] The peptides can be used to design innovative specific local treatments for diseases that depend on IL-17A overproduction or activity.

[0024] The present invention relates to peptides, dimers and bioconjugates capable of inhibiting the binding of IL-17A to IL-17RA, pharmaceutical compositions comprising the peptides, dimers or bioconjugates, and the use of the above substances in preventing and / or treating autoimmune diseases and inflammatory diseases that are dependent on IL-17A. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 Shown are some dose response curves obtained using representative IL-17A binding peptides according to the invention in the IL-17RA-IL17RC dimerization assay described in Example 5. The percentage inhibition of IL-17RA / IL17RC dimerization induced by IL-17A obtained by treatment with the different peptides is reported.

[0027] Figure 2 The activity of the IL-17A binding peptide of SEQ ID N.18 on the secretion of IL6, IL-17A and IL23 by human differentiated and activated TH17 cells is shown, measured as described in Example 6. The concentrations of the different cytokines in the supernatant of TH17 cells with or without the peptide of SEQ ID N.18 (control) are reported.

[0028] Figure 3 The activity of the IL-17A binding peptide of SEQ ID N.18 on the secretion of metalloproteinase 3 by human differentiated and activated TH17 cells is shown, measured as described in Example 6. The concentration of metalloproteinase 3 in the supernatant of TH17 cells with or without the peptide of SEQ ID N.18 (control) is reported.

[0029] Figure 4 The viability of human TH17 cells after 24 h in the presence and absence (control) of the peptide of SEQ ID N 18 as described in Example 6 is shown.

[0030] Figure 5 The binding site of the IL-17A peptide according to the present invention on IL-17A is shown.

[0031] Figure 6 Sensorgrams obtained in the experiments of Example 9 using the peptide of SEQ ID N 2 (upper panel) or SEQ ID N 1 (lower panel) are shown.

[0032] Figure 7 The values ​​of affinity constants for different concentrations of the peptide SEQ ID N2 (upper graph) or SEQ ID N1 (lower graph) obtained in the experiment of Example 9 are shown.

[0033] Figure 8 The expression level of IL-8 in HCEC cells untreated (vehicle) or treated with the peptide of SEQ ID N.2 (A) or treated with the peptide HAP (SEQ ID N.1) (B) measured by real-time PCR as described in Example 10 is shown. Error bars represent standard deviations. T-test was performed as statistical analysis. P values ​​*<0.05, **<0.005, ***<0.0005.

[0034] Fig. 9 The amount of IL-6 in HCEC cells untreated (vehicle) or treated with the peptide SEQ ID N.2 (Panel A) or treated with the peptide HAP (SEQ ID N.1) (Panel B) measured by real-time PCR as described in Example 10 is shown. Error bars represent standard deviations. T-tests were performed as statistical analysis. P values ​​*<0.05, **<0.005, ***<0.0005.

[0035] Fig.10 The amount of TNF-α in HCEC cells untreated (vehicle) or treated with the peptide of SEQ ID N.2 (Panel A) or treated with the peptide HAP (SEQ ID N.1) (Panel B) measured by real-time PCR as described in Example 10 is shown. Error bars represent standard deviations. T-test was performed as statistical analysis. P values ​​*<0.05, **<0.005, ***<0.0005.

[0036] definition

[0037] Unless otherwise defined, all technical terms, symbols and other scientific terms used herein are intended to have the meanings commonly understood by those skilled in the art to which the present disclosure belongs. In some cases, terms with commonly understood meanings are defined herein for clarity and / or ease of reference; therefore, the inclusion of such definitions herein should not be interpreted as having a substantial difference from the meanings generally understood in the art.

[0038] The term "pharmaceutically acceptable excipient" refers to a substance other than an active pharmaceutical ingredient (API) that has been evaluated for safety and is intended to be included in a drug delivery system. Pharmaceutically acceptable excipients are well known in the art and are disclosed, for example, in Handbook of Pharmaceutical Excipients (Seventh Edition, 2013), which is incorporated herein by reference (Rowe, Sheskey et al. 2012).

[0039] Excipients are usually classified according to their function in the final pharmaceutical composition. Preferably, suitable excipients according to the present invention are, for example, diluents, adsorbents, glidants, binders, lubricants, surfactants, disintegrants, preservatives, antioxidants or mixtures thereof.

[0040] The terms "about" and "approximately" herein refer to the range of experimental errors that may occur in the measurements.

[0041] The terms "comprising," "having," "including," and "containing" should be construed as open-ended terms (ie, meaning "including but not limited to"), and should be considered to also provide support for the terms "consisting essentially of" or "consisting of.

[0042] The term "consisting essentially of" should be interpreted as a semi-closed term, meaning that other ingredients (and therefore optional excipients) that materially affect the basic and novel characteristics of the invention are not included.

[0043] The term "consisting of" should be interpreted as a closed term.

[0044] As used herein, the term "bioconjugate" refers to a conjugate formed by a stable covalent linkage between different molecules, preferably between two molecules, optionally linked via a spacer, at least one of which is a biomolecule.

[0045] As used herein, the term "biomolecule" refers to a molecule of biological origin. The term includes macromolecules such as carbohydrates, lipids and proteins or small natural products. For the purposes of the present invention, "biomolecule" is preferably selected from ascorbic acid, capric acid, caproic acid, N-acetylglucosamine (also known as NAG), N-acetylmuramic acid (also known as NAM), hyaluronic acid, alginic acid, chitin, (GalNAc)2, Gal-α1, 3-GalNAc or trigalacturonic acid.

[0046] As defined herein, "conservative substitutions" refer to conservative replacements (also called conservative mutations or conservative substitutions), which are amino acid replacements that change a given amino acid to a different amino acid with similar biochemical properties (Simon French 1983). DETAILED DESCRIPTION OF THE INVENTION

[0048] The present inventors have identified a number of peptides that are able to bind to IL-17A with high affinity and inhibit its interaction with the ILRA receptor. These compounds act as inhibitors of IL-17A signal transduction.

[0049] Therefore, the first object of the present invention is a peptide capable of inhibiting the binding of IL-17A to ILRA, which has the following amino acid sequence:

[0050] Formula (I):

[0051] (I)X1-X2-X3-X4-X5-X6-X7-X8-X9-X 10 -X 11 -X 12 -X 13 -X 14 -X 15

[0052] Among them, independently of each other:

[0053] X1 is I, V, D, K, W, A, G, L, or P;

[0054] X2 is H, M, K, N, R, E, Q, W, or Y;

[0055] X3 is V, F, A, G, L, P, I, Y, or W;

[0056] X4 is T, Q, H, S, N or Y;

[0057] X5 is I, F, A, G, L, P, V, or Y;

[0058] X6 is P, G, A, L, V, I or N;

[0059] X7 is A, Q, G, L, P, V, I, N, or E;

[0060] X8 is D, E, N, Q or Y;

[0061] X9 is L, V, F, W, A, G, P, I or H;

[0062] X 10 is W, Y, F or Q or does not exist;

[0063] X 11 is D, E or N or does not exist;

[0064] X 12 is W, F, V, H or Y or does not exist;

[0065] X 13 is I, V, F, E, K, A, G, L, P or Y or is absent;

[0066] X 14 is N, R, E, F, Q or D or is absent;

[0067] X 15 is K, R, E, F, V, W, H or D or does not exist;

[0068] Provided that the sequence is not IHVTIPADLWDWINK (SEQ ID N.1) or its 12-14 amino acid long C-terminal truncated sequence IHVTIPADLWDWIN, IHVTIPADLWDWI or IHVTIPADLWDW;

[0069] Or formula (II):

[0070] (II)a n -DLSAVCWAFPWDPECH-b n’

[0071] Among them, independently of each other:

[0072] a, b are selected from A, R, N, D, C, E, Q, G, H, I, L, K, M, F, P, S, T, W, Y or V;

[0073] n=an integer from 0 to 3;

[0074] n′=an integer from 0 to 3.

[0075] Preferably, in formula (II), each amino acid that differs from a or b may be replaced by a conservative substitution.

[0076] A preferred peptide of formula (II) is DLSAVCWAFPWDPECH (SEQ ID N. 41).

[0077] Particularly preferred are peptides of formula (I).

[0078] According to a preferred embodiment, in the peptide of formula (I):

[0079] X1 is I, V, D, K, or W;

[0080] X2 is H, M, K or N;

[0081] X3 is V or F;

[0082] X4 is T, Q, H, V or Y;

[0083] X5 is I or F;

[0084] X6 is P or G;

[0085] X7 is A or Q;

[0086] X8 is D, E, N;

[0087] X9 is L, V, F or W;

[0088] X 10 is W, Y, F or does not exist;

[0089] X11 is D, E, N or does not exist;

[0090] X 12 is W, F, V or absent;

[0091] X 13 is I, V, F or absent;

[0092] X 14 is N, R, E, F or absent;

[0093] X 15 is K, R, E, F, V, W or not present;

[0094] The proviso is that the sequence is not IHVTIPADLWDWINK (SEQ ID N. 1) or its 12-14 amino acid long C-terminal truncated sequence IHVTIPADLWDWIN, IHVTIPADLWDWI or IHVTIPADLWDW.

[0095] According to an alternative preferred embodiment, in the peptide of formula (I):

[0096] X1 is I, V, A, G, L, or P;

[0097] X2 is H, M, K, R, E, Q, W, or Y;

[0098] X3 is V, F, A, G, L, P, I, Y, or W;

[0099] X4 is T, Q, S, N or Y;

[0100] X5 is I, F, A, G, L, P, V, or Y;

[0101] X6 is P, G, A, L, V, I or N;

[0102] X7 is A, Q, G, L, P, V, I, N, or E;

[0103] X8 is D, E, N, Q or Y;

[0104] X9 is L, V, F, W, A, G, P, I or H;

[0105] X 10 is W, Y, F or Q or does not exist;

[0106] X 11 is D, E or N or does not exist;

[0107] X 12 is W, F, H or Y or does not exist;

[0108] X 13is I, V, F, E, K, A, G, L, P or Y or is absent;

[0109] X 14 is N, R, E, Q or D or does not exist;

[0110] X 15 is K, R, E, V, W, H or D or does not exist;

[0111] The proviso is that the sequence is not IHVTIPADLWDWINK (SEQ ID N. 1) or its 12-14 amino acid long C-terminal truncated sequence IHVTIPADLWDWIN, IHVTIPADLWDWI or IHVTIPADLWDW.

[0112] Preferably, in the peptide of formula (I) above, all amino acids X1 to X 15 , and the peptide of formula (I) above has a sequence of 15 amino acids. Preferably, in a peptide of formula (I) having a sequence of less than 15 amino acids, when an amino acid is absent, all amino acids at the N-terminus of the amino acid are also absent. For example, if amino acid X 10 If not present, then amino acid X 11 To X 15 Doesn't exist either.

[0113] Particularly preferred peptides of formula (I) are those wherein X1 is V.

[0114] A particularly preferred peptide having X1 as V is the peptide of SEQ ID N2.

[0115] As demonstrated in the experimental part, the inventors found that, surprisingly, the peptide of SEQ ID N 2, in which the first amino acid isoleucine in the peptide HAP of SEQ ID N.1 has been replaced by valine, shows significantly different functional and chemical-physical properties and improved tolerability compared to HAP.

[0116] In particular, as demonstrated in the examples, the peptide exhibits improved chemical-physical properties that are particularly advantageous for topical and ophthalmic use, with better permeability and enhanced affinity for IL-17A compared to the corresponding peptide of SEQ ID N 1. In addition, the inventors have found that the peptide HAP exerts a direct toxic effect on corneal cells by inducing the expression of inflammatory cytokines in a manner independent of its activity on IL-17A signaling. Surprisingly, the peptide of SEQ ID N.2 does not show this effect and is therefore characterized by an improved tolerability compared to HAP.

[0117] Preferably, in the peptide of the above formula (I):

[0118] X1 is I, V or L;

[0119] X2 is H, M, R, K or E;

[0120] X3 is V, F or I;

[0121] X4 is T, Q, S, Y or N;

[0122] X5 is I, F, or V;

[0123] X6 is P;

[0124] X7 is A, Q or L;

[0125] X8 is D, E or Q;

[0126] X9 is L, W, F, V, or I;

[0127] X 10 is W, Y or F;

[0128] X 11 is D, E or N;

[0129] X 12 is W or F;

[0130] X 13 is I, V, F, or L;

[0131] X 14 is N, R, Q or E;

[0132] X 15 is K, R, H or E;

[0133] Provided that the sequence is not IHVTIPADLWDWINK (SEQ ID N.1);

[0134] or

[0135] X1 is I or V;

[0136] X2 is H, M or R;

[0137] X3 is V or F;

[0138] X4 is T or Q;

[0139] X5 is I, F, or V;

[0140] X6 is P or G;

[0141] X7 is A or Q;

[0142] X8 is D or E;

[0143] X9 is L;

[0144] X 10 is W or Y;

[0145] X 11 is D or E;

[0146] X 12 It is W;

[0147] X 13 is I or V;

[0148] X 14 is N, R or E;

[0149] X 15 is K, R or E;

[0150] Provided that the sequence is not IHVTIPADLWDWINK (SEQ ID N.1);

[0151] or

[0152] X1 is I or V;

[0153] X2 is H or M;

[0154] X3 is V;

[0155] X4 is T;

[0156] X5 is I;

[0157] X6 is P;

[0158] X7 is A;

[0159] X8 is D;

[0160] X9 is L, W, F, V, or I;

[0161] X 10 is W or Y;

[0162] X 11 is D or E;

[0163] X 12 It is W;

[0164] X 13 is I or V;

[0165] X 14 is N, R or E;

[0166] X 15 is K, R or E;

[0167] Provided that the sequence is not IHVTIPADLWDWINK (SEQ ID N.1);

[0168] or

[0169] X1 is I, V, A, G, L, or P;

[0170] X2 is M, K, R, E, Q, W, or Y;

[0171] X3 is V, F, A, G, L, P, I, Y, or W;

[0172] X4 is T, Q, S, N or Y;

[0173] X5 is I, F, A, G, L, P, V, or Y;

[0174] X6 is P, G, A, L, V, I or N;

[0175] X7 is A, Q, G, L, P, V, I, N, or E;

[0176] X8 is D, E, N, Q or Y;

[0177] X9 is L, V, F, W, A, G, P, I or H;

[0178] X 10 is Y, F or Q;

[0179] X 11 is D, E or N;

[0180] X 12 is W, F, H or Y;

[0181] X 13 It is I, V, F, E, K, A, G, L, P;

[0182] X 14 is N, R, E, Q or D;

[0183] X 15 is K, R, E, V, W, H or D;

[0184] or

[0185] X1 is V, A, G, L, or P;

[0186] X2 is H, M, K, R, E, Q, W, or Y;

[0187] X3 is V, F, A, G, L, P, I, Y, or W;

[0188] X4 is T, Q, S, N or Y;

[0189] X5 is I, F, A, G, L, P, V, or Y;

[0190] X6 is P, G, A, L, V, I or N;

[0191] X7 is A, Q, G, L, P, V, I, N, or E;

[0192] X8 is D, E, N, Q or Y;

[0193] X9 is L, V, F, W, A, G, P, I or H;

[0194] X 10 is Y, F or Q;

[0195] X 11 is D, E or N;

[0196] X 12 is W, F, H or Y;

[0197] X 13 is I, V, F, E, K, A, G, L, P or Y;

[0198] X 14 is N, R, E, Q or D;

[0199] X 15 is K, R, E, V, W, H or D;

[0200] or

[0201] X1 is V, A, G, L, or P;

[0202] X2 is M, K, R, E, Q, W, or Y;

[0203] X3 is V, F, A, G, L, P, I, Y, or W;

[0204] X4 is T, Q, S, N or Y;

[0205] X5 is I, F, A, G, L, P, V, or Y;

[0206] X6 is P, G, A, L, V, I or N;

[0207] X7 is A, Q, G, L, P, V, I, N, or E;

[0208] X8 is D, E, N, Q or Y;

[0209] X9 is L, V, F, W, A, G, P, I or H;

[0210] X 10 is Y, F or Q;

[0211] X 11 is D, E or N;

[0212] X 12 is W, F, H or Y;

[0213] X 13 is I, V, F, E, K, A, G, L, P or Y;

[0214] X 14 is N, R, E, Q or D;

[0215] X 15 is K, R, E, V, W, H or D;

[0216] or

[0217] X1 is V, A, G, L, or P;

[0218] X2 is H, M, K, R, E, Q, W, or Y;

[0219] X3 is V, F, A, G, L, P, I, Y, or W;

[0220] X4 is T, Q, S, N or Y;

[0221] X5 is I, F, A, G, L, P, V, or Y;

[0222] X6 is P, G, A, L, V, I or N;

[0223] X7 is A, Q, G, L, P, V, I, N, or E;

[0224] X8 is D, E, N, Q or Y;

[0225] X9 is L, V, F, W, A, G, P, I or H;

[0226] X 10 is W, Y, F or Q;

[0227] X 11 is D, E or N;

[0228] X 12 is W, F, H or Y;

[0229] X 13 is V, F, E, K, A, G, L, P or Y;

[0230] X 14 is N, R, E, Q or D;

[0231] X 15 is K, R, E, V, W, H or D;

[0232] or

[0233] X1 is I, V, A, G, L, or P;

[0234] X2 is M, K, R, E, Q, W, or Y;

[0235] X3 is V, F, A, G, L, P, I, Y, or W;

[0236] X4 is T, Q, S, N or Y;

[0237] X5 is I, F, A, G, L, P, V, or Y;

[0238] X6 is P, G, A, L, V, I or N;

[0239] X7 is A, Q, G, L, P, V, I, N, or E;

[0240] X8 is D, E, N, Q or Y;

[0241] X9 is L, V, F, W, A, G, P, I or H;

[0242] X 10 is W, Y, F or Q;

[0243] X 11 is D, E or N;

[0244] X 12 is W, F, H or Y;

[0245] X 13 is V, F, E, K, A, G, L, P or Y;

[0246] X 14 is N, R, E, Q or D;

[0247] X 15 is K, R, E, V, W, H or D;

[0248] or

[0249] X1 is I, V, A, G, L, or P;

[0250] X2 is H, M, K, R, E, Q, W, or Y;

[0251] X3 is V, F, A, G, L, P, I, Y, or W;

[0252] X4 is T, Q, S, N or Y;

[0253] X5 is I, F, A, G, L, P, V, or Y;

[0254] X6 is P, G, A, L, V, I or N;

[0255] X7 is A, Q, G, L, P, V, I, N, or E;

[0256] X8 is D, E, N, Q or Y;

[0257] X9 is L, V, F, W, A, G, P, I or H;

[0258] X 10 is Y, F or Q;

[0259] X 11 is D, E or N;

[0260] X 12 is W, F, H or Y;

[0261] X 13 is V, F, E, K, A, G, L, P or Y;

[0262] X 14 is N, R, E, Q or D;

[0263] X 15 is K, R, E, V, W, H or D;

[0264] or

[0265] X1 is I, V, A, G, L, or P;

[0266] X2 is M, K, R, E, Q, W, or Y;

[0267] X3 is V, F, A, G, L, P, I, Y, or W;

[0268] X4 is T, Q, S, N or Y;

[0269] X5 is I, F, A, G, L, P, V, or Y;

[0270] X6 is P, G, A, L, V, I or N;

[0271] X7 is A, Q, G, L, P, V, I, N, or E;

[0272] X8 is D, E, N, Q or Y;

[0273] X9 is L, V, F, W, A, G, P, I or H;

[0274] X 10 is Y, F or Q;

[0275] X 11 is D, E or N;

[0276] X 12 is W, F, H or Y;

[0277] X 13 is V, F, E, K, A, G, L, P or Y;

[0278] X 14is N, R, E, Q or D;

[0279] X 15 is K, R, E, V, W, H or D;

[0280] or

[0281] X1 is V, A, G, L, or P;

[0282] X2 is H, M, K, R, E, Q, W, or Y;

[0283] X3 is V, F, A, G, L, P, I, Y, or W;

[0284] X4 is T, Q, S, N or Y;

[0285] X5 is I, F, A, G, L, P, V, or Y;

[0286] X6 is P, G, A, L, V, I or N;

[0287] X7 is A, Q, G, L, P, V, I, N, or E;

[0288] X8 is D, E, N, Q or Y;

[0289] X9 is L, V, F, W, A, G, P, I or H;

[0290] X 10 is Y, F or Q;

[0291] X 11 is D, E or N;

[0292] X 12 is W, F, H or Y;

[0293] X 13 is V, F, E, K, A, G, L, P or Y;

[0294] X 14 is N, R, E, Q or D;

[0295] X 15 is K, R, E, V, W, H or D;

[0296] or

[0297] X1 is V, A, G, L, or P;

[0298] X2 is M, K, R, E, Q, W, or Y;

[0299] X3 is V, F, A, G, L, P, I, Y, or W;

[0300] X4 is T, Q, S, N or Y;

[0301] X5 is I, F, A, G, L, P, V, or Y;

[0302] X6 is P, G, A, L, V, I or N;

[0303] X7 is A, Q, G, L, P, V, I, N, or E;

[0304] X8 is D, E, N, Q or Y;

[0305] X9 is L, V, F, W, A, G, P, I or H;

[0306] X 10 is Y, F or Q;

[0307] X 11 is D, E or N;

[0308] X 12 is W, F, H or Y;

[0309] X 13 is V, F, E, K, A, G, L, P or Y;

[0310] X 14 is N, R, E, Q or D;

[0311] X 15 is K, R, E, V, W, H or D;

[0312] or

[0313] X1 is I, V, A, G, L, or P;

[0314] X2 is M, K, N, R, E, Q, W, or Y;

[0315] X3 is V, F, A, G, L, P, I, Y, or W;

[0316] X4 is T, Q, S, N or Y;

[0317] X5 is I, F, A, G, L, P, V, or Y;

[0318] X6 is P, G, A, L, V, I or N;

[0319] X7 is A, Q, G, L, P, V, I, N, or E;

[0320] X8 is D, E, N, Q or Y;

[0321] X9 is L, V, F, W, A, G, P, I or H;

[0322] X 10 is W, Y, F or Q;

[0323] X 11 is E or N;

[0324] X 12 is W, F, H or Y;

[0325] X 13 is I, V, F, E, K, A, G, L, P or Y;

[0326] X 14 is N, R, E, Q or D;

[0327] X 15 is K, R, E, V, W, H or D;

[0328] or

[0329] X1 is I, V, A, G, L, or P;

[0330] X2 is H, M, K, N, R, E, Q, W, or Y;

[0331] X3 is V, F, A, G, L, P, I, Y, or W;

[0332] X4 is T, Q, S, N or Y;

[0333] X5 is I, F, A, G, L, P, V, or Y;

[0334] X6 is P, G, A, L, V, I or N;

[0335] X7 is A, Q, G, L, P, V, I, N, or E;

[0336] X8 is D, E, N, Q or Y;

[0337] X9 is L, V, F, W, A, G, P, I or H;

[0338] X 10 is W, Y, F or Q;

[0339] X 11 is E or N;

[0340] X 12 is W, F, H or Y;

[0341] X 13 is I, V, F, E, K, A, G, L, P or Y;

[0342] X 14 is N, R, E, Q or D;

[0343] X 15 is K, R, E, V, W, H or D;

[0344] or

[0345] X1 is V, A, G, L, or P;

[0346] X2 is M, K, N, R, E, Q, W, or Y;

[0347] X3 is V, F, A, G, L, P, I, Y, or W;

[0348] X4 is T, Q, S, N or Y;

[0349] X5 is I, F, A, G, L, P, V, or Y;

[0350] X6 is P, G, A, L, V, I or N;

[0351] X7 is A, Q, G, L, P, V, I, N, or E;

[0352] X8 is D, E, N, Q or Y;

[0353] X9 is L, V, F, W, A, G, P, I or H;

[0354] X 10 is W, Y, F or Q;

[0355] X 11 is E or N;

[0356] X 12 is W, F, H or Y;

[0357] X 13 is I, V, F, E, K, A, G, L, P or Y;

[0358] X 14 is N, R, E, Q or D;

[0359] X 15 is K, R, E, V, W, H or D;

[0360] or

[0361] X1 is I, V, A, G, L, or P;

[0362] X2 is H, M, K, R, E, Q, W, or Y;

[0363] X3 is V, F, A, G, L, P, I, Y, or W;

[0364] X4 is T, Q, S, N or Y;

[0365] X5 is I, F, A, G, L, P, V, or Y;

[0366] X6 is P, G, A, L, V, I or N;

[0367] X7 is A, Q, G, L, P, V, I, N, or E;

[0368] X8 is D, E, N, Q or Y;

[0369] X9 is L, V, F, W, A, G, P, I or H;

[0370] X 10 is Y, F or Q;

[0371] X 11 is E or N;

[0372] X 12 is W, F, H or Y;

[0373] X 13 is I, V, F, E, K, A, G, L, P or Y;

[0374] X 14 is N, R, E, Q or D;

[0375] X 15 is K, R, E, V, W, H or D;

[0376] or

[0377] X1 is I, V, A, G, L, or P;

[0378] X2 is H, M, K, R, E, Q, W, or Y;

[0379] X3 is V, F, A, G, L, P, I, Y, or W;

[0380] X4 is T, Q, S, N or Y;

[0381] X5 is I, F, A, G, L, P, V, or Y;

[0382] X6 is P, G, A, L, V, I or N;

[0383] X7 is A, Q, G, L, P, V, I, N, or E;

[0384] X8 is D, E, N, Q or Y;

[0385] X9 is L, V, F, W, A, G, P, I or H;

[0386] X 10 is W, Y, F or Q;

[0387] X 11 is E or N;

[0388] X 12is W, F, H or Y;

[0389] X 13 is V, F, E, K, A, G, L, P or Y;

[0390] X 14 is N, R, E, Q or D;

[0391] X 15 is K, R, E, V, W, H or D;

[0392] or

[0393] X1 is I, V, A, G, L, or P;

[0394] X2 is H, M, K, R, E, Q, W, or Y;

[0395] X3 is V, F, A, G, L, P, I, Y, or W;

[0396] X4 is T, Q, S, N or Y;

[0397] X5 is I, F, A, G, L, P, V, or Y;

[0398] X6 is P, G, A, L, V, I or N;

[0399] X7 is A, Q, G, L, P, V, I, N, or E;

[0400] X8 is D, E, N, Q or Y;

[0401] X9 is L, V, F, W, A, G, P, I or H;

[0402] X 10 is Y, F or Q;

[0403] X 11 is E or N;

[0404] X 12 is W, F, H or Y;

[0405] X 13 is V, F, E, K, A, G, L, P or Y;

[0406] X 14 is N, R, E, Q or D;

[0407] X 15 is K, R, E, V, W, H or D;

[0408] or

[0409] X1 is I, V, A, G, L, or P;

[0410] X2 is H, M, K, R, E, Q, W, or Y;

[0411] X3 is V, F, A, G, L, P, I, Y, or W;

[0412] X4 is T, Q, S, N or Y;

[0413] X5 is I, F, A, G, L, P, V, or Y;

[0414] X6 is P, G, A, L, V, I or N;

[0415] X7 is A, Q, G, L, P, V, I, N, or E;

[0416] X8 is D, E, N, Q or Y;

[0417] X9 is L, V, F, W, A, G, P, I or H;

[0418] X 10 is W, Y, F or Q;

[0419] X 11 is D, E or N;

[0420] X 12 is W, F, H or Y;

[0421] X 13 is I, V, F, E, K, A, G, L, P or Y;

[0422] X 14 is R, E, Q or D;

[0423] X 15 is R, E, V, W, H or D;

[0424] or

[0425] X1 is V, A, G, L, or P;

[0426] X2 is H, M, K, R, E, Q, W, or Y;

[0427] X3 is V, F, A, G, L, P, I, Y, or W;

[0428] X4 is T, Q, S, N or Y;

[0429] X5 is I, F, A, G, L, P, V, or Y;

[0430] X6 is P, G, A, L, V, I or N;

[0431] X7 is A, Q, G, L, P, V, I, N, or E;

[0432] X8 is D, E, N, Q or Y;

[0433] X9 is L, V, F, W, A, G, P, I or H;

[0434] X 10 is W, Y, F or Q;

[0435] X 11 is D, E or N;

[0436] X 12 is W, F, H or Y;

[0437] X 13 is I, V, F, E, K, A, G, L, P or Y;

[0438] X 14 is R, E, Q or D;

[0439] X 15 is R, E, V, W, H or D;

[0440] or

[0441] X1 is I, V, A, G, L, or P;

[0442] X2 is M, K, R, E, Q, W, or Y;

[0443] X3 is V, F, A, G, L, P, I, Y, or W;

[0444] X4 is T, Q, S, N or Y;

[0445] X5 is I, F, A, G, L, P, V, or Y;

[0446] X6 is P, G, A, L, V, I or N;

[0447] X7 is A, Q, G, L, P, V, I, N, or E;

[0448] X8 is D, E, N, Q or Y;

[0449] X9 is L, V, F, W, A, G, P, I or H;

[0450] X 10 is W, Y, F or Q;

[0451] X 11 is D, E or N;

[0452] X 12 is W, F, H or Y;

[0453] X 13 is I, V, F, E, K, A, G, L, P or Y;

[0454] X 14 is R, E, Q or D;

[0455] X 15 Is R, E, V, W, H or D.

[0456] Particularly preferred peptides of formula (I) among those mentioned above are those wherein X1 is V.

[0457] As demonstrated in the experimental part, when the first amino acid in the peptide of formula (I) is valine, the peptide surprisingly exhibits particularly advantageous chemicophysical properties for topical and ophthalmic use and an increased affinity for IL-17A.

[0458] Preferably, in the peptide of formula (I) above, all amino acids X1 to X 15 .

[0459] Preferred individual peptides of formula (I) according to the invention are listed in Table 1 below.

[0460] Table 1

[0461]

[0462]

[0463]

[0464]

[0465]

[0466] Other peptide inhibitors of IL-17A according to the invention are those of Table 1A.

[0467] Table 1A:

[0468] SEQ ID N.156 VPGAGVPGAGIHVTI SEQ ID N.157 VPGAGVPGAGIHVTIPA

[0469] Other amino acid sequences may be added to the N-terminus or C-terminus of the peptide according to the first object of the present invention.

[0470] According to a preferred embodiment, the peptide of formula (I) or formula (II) is bound at its N-terminus and / or C-terminus to another amino acid sequence, namely sequence A, which has an amino acid sequence of formula (III)

[0471] (III)Y1-Y2-Y3-Y4-Y5-Y6-Y7-Y8-Y9-Y 10 -Y 11 -Y 12- Y 13 -Y 14 -Y15 -Y 16

[0472] in:

[0473] Y1 is A, T, V, K, R, I, L, X, or G;

[0474] Y2 is R, W, P, E, Q or A;

[0475] Y3 is K, W, G, T, I, R or P;

[0476] Y4 is K, T, E, W, A, R, D, G, X or F;

[0477] Y5 is A, E, T, G, W, I, R, P or V;

[0478] Y6 is A, W, E, R, G, P or absent;

[0479] Y7 is K, S, W, T, F, R, V, G or absent;

[0480] Y8 is A, Q, W, R, G or absent;

[0481] Y9 is N, G or absent;

[0482] Y 10 is R or not present;

[0483] Y 11 is R or not present;

[0484] Y 12 It is M or not present;

[0485] Y 13 It is K or not present;

[0486] Y 14 It is W or not present;

[0487] Y 15 It is K or not present;

[0488] Y 16 It is K or not present.

[0489] Preferably, in the sequence of formula (III), when an amino acid is absent, the amino acid at the N-terminus of the amino acid is also absent. For example, if amino acid Y6 is absent, then amino acids Y7 to Y 16 Doesn't exist either.

[0490] Preferred sequences of formula (III) according to the present invention are listed in Table 2 below.

[0491] Table 2

[0492] Sequence ID sequence SEQ ID N.44 ARKKAAKA SEQ ID N.45 TWWTEWSQ SEQ ID N.46 TWWETWW SEQ ID N.47 VPGWG SEQ ID N.48 KETWWETW SEQ ID N.49 VPGK SEQ ID N.50 VPGAG SEQ ID N.51 RQIKIWFQNRRMKWKK SEQ ID N.52 RRRRRRRR SEQ ID N.53 VPGDG SEQ ID N.54 VPGXG SEQ ID N.55 IPGG SEQ ID N.56 AVGVP SEQ ID N.57 IPGXG SEQ ID N.58 IPGVG SEQ ID N.59 LPGXG SEQ ID N.60 LPGVG SEQ ID N.61 VAPGVG SEQ ID N.62 XPG SEQ ID N.63 GVGVPGVG SEQ ID N.64 VPGFGVGAG SEQ ID N.65 VPGGVPGG

[0493] Therefore, the second object of the present invention is a peptide having the following amino acid sequence, which contains, preferably consists of, the peptide sequence of formula (I) or formula (II) according to the first object of the present invention and the sequence A as described above at the C-terminus and / or N-terminus of the sequence.

[0494] The amino acid sequence A may be bound to the N-terminus and the C-terminus of the peptide of formula (I) or formula (II). In this case, the two amino acid sequences A bound to the N-terminus and the C-terminus of the peptide may be the same or different.

[0495] Alternatively, the amino acid sequence A may be bound to the N-terminus or C-terminus of the peptide.

[0496] Preferred individual peptides for this purpose according to the invention are listed in Table 3 below.

[0497] Table 3

[0498]

[0499]

[0500] In the above alternative embodiments, the peptide of formula (I) or formula (II) according to the first object of the present invention may be bound at its N-terminus and / or C-terminus to the amino acid sequence (A) m , wherein A is an amino acid selected from R, K, G, E, Q or A, preferably R, K, G, E or A, and m is an integer from 1 to 10.

[0501] Amino acid sequence (A) m It can be bound to the N-terminus and C-terminus of the peptide of formula (I) or formula (II). In this case, the two amino acid sequences (A) connected to the N-terminus and C-terminus of the peptide are m Can be the same or different.

[0502] Alternatively, the amino acid sequence (A) m Linkage may be to the N-terminus or the C-terminus of the peptide.

[0503] Therefore, a third object of the present invention is a peptide having an amino acid sequence comprising, preferably consisting of, the peptide sequence according to the first object of the present invention and a sequence (A) as defined above at the C-terminus and / or N-terminus of said sequence: m .

[0504] Preferred individual peptides according to this embodiment of the invention are listed in Table 4 below.

[0505] Table 4

[0506] SEQ ID N. sequence SEQ ID N.19 VMVTIPADLYEWIEERRRRRR SEQ ID N.20 VMVTIPADLYEWIEERRR SEQ ID N.29 RRRRRRRRVMVTIPADLYEWIEE SEQ ID N.30 VMVTIPADLYEWIEERRRRRRRR SEQ ID N.43 IMVTIPADLYEWIEEQ

[0507] In a specific embodiment, the peptide of formula (I) or formula (II) is a peptide of sequence A and sequence (A) m In this case, the peptide is bound at its N-terminus and / or C-terminus to an amino acid sequence (A) as defined above. m , the amino acid sequence (A) m It is bound to the amino acid sequence A as described above at its N-terminus and / or C-terminus.

[0508] Therefore, the fourth object of the present invention is a peptide having an amino acid sequence, which comprises, preferably consists of, the peptide sequence according to the first object of the present invention, the sequence (A) as described above at the C-terminus and / or N-terminus of the sequence m , and in the sequence (A) m The C-terminus and / or N-terminus of sequence A as described above.

[0509] According to a preferred embodiment of the peptide according to any of the above objects of the invention, the peptide is bound at the C-terminus and / or N-terminus to a protective cap group capable of preventing degradation.

[0510] Any protective capping group conventionally used in the art may be used.

[0511] Preferably, the protective cap group bound to the C-terminus is selected from amides, preferably N-alkylamides, aldehydes, esters, preferably methyl and ethyl esters, p-nitroaniline, 7-amino-4-methylcoumarin.

[0512] Preferably, the protective capping group bound to the N-terminus is selected from acetyl, formyl, pyroglutamyl, fatty acid, urea, carbamate sulfonamide, alkylamine.

[0513] Preferably, the above peptide is bound to a protective cap group only at the N-terminus. More preferably, the peptide is bound to a protective cap group only at the N-terminus, and the protective cap group is an acetyl group.

[0514] It is particularly advantageous to prepare dimers of the above peptides according to any of the different objects of the invention, preferably dimers of the peptides of formula (I) or formula (II) according to the first object of the invention, in order to increase binding affinity and inhibitor activity.

[0515] According to the above, preferably, the peptides according to the different purposes of the present invention, preferably the peptides of formula (I) or formula (II) according to the first purpose of the present invention, more preferably the peptides of formula (I) according to the first purpose of the present invention exist in the form of dimers.

[0516] Therefore, the fifth object of the present invention is a dimer formed by two peptides according to any of the above objects of the present invention. Preferably, the peptides are peptides of formula (I) or formula (II) according to the first object of the present invention, more preferably, they are peptides of formula (I) according to the first object of the present invention.

[0517] Preferably, the dimer is a homodimer, wherein the two peptides forming the dimer have the same sequence.Preferably, in the dimer, the peptides are linked via a spacer molecule, preferably a polyethylene glycol spacer.

[0518] Preferably, according to this embodiment, polyethylene glycol is linked to the N-terminus of the peptide according to the invention or to two amino acids X4, X7 or X8 of the peptide of formula (I). 14 .

[0519] According to another embodiment, the peptides according to the different purposes of the present invention are preferably bound to a biomolecule at its C-terminus and / or N-terminus to form a bioconjugate in the absence of a protective group cap. Preferably, the biomolecule is selected from the group consisting of capric acid, caproic acid, ascorbic acid, NAG-NAM, NAG, NAM, hyaluronic acid, alginic acid, chitin, (GalNAc)2, Gal-α1, 3-GalNAc and trigalacturonic acid.

[0520] Biomolecules are conjugated to peptides to add specific functions to them or to modulate their physicochemical properties. For example, ascorbic acid provides antioxidant activity to peptides, and hexanoic acid helps anchor peptides to cell membranes.

[0521] Therefore, a sixth object of the present invention is a bioconjugate comprising, preferably consisting of, a peptide according to a different object of the present invention, preferably a peptide of formula (I) or formula (II) according to the first object of the present invention, more preferably a peptide of formula (I) according to the first object of the present invention, and at least one biomolecule as described above.

[0522] Preferably, in the bioconjugate, the biomolecule is bound to the N-terminus and / or C-terminus of the peptide.

[0523] Preferred bioconjugates according to this embodiment are listed in Table 5 below.

[0524] Table 5

[0525] Name of bioconjugate sequence Bio-1 NAM-IMVTIPADLYEWIEE Bio-2 IMVTIPADLYEWIEEQ-NAG Bio-3 NAG-NAM-IMVTIPADLYEWIEE Bio-4 Decanoic acid-IMVTIPADLYEWIEE Bio-5 Ascorbic acid -IMVTIPADLYEWIEE

[0526] According to one embodiment, in the above bioconjugate, the above biomolecule or protective cap group is directly bound to the N-terminus and / or C-terminus of the peptide.

[0527] According to one embodiment, the biomolecule is bound to the N-terminus and the C-terminus of the peptide. In such cases, the two biomolecules bound to the N-terminus and the C-terminus of the peptide may be the same or different.

[0528] According to an alternative embodiment, the biomolecule is bound to the N-terminus or the C-terminus of the peptide.

[0529] According to alternative embodiments, the above-mentioned biomolecule or protective cap group is connected to the N-terminus and / or C-terminus of the peptide via a linker. Preferably, the linker is selected from 4-aminobutyric acid, β-alanine, 2-aminoethoxy-acetic acid, 5-aminopentanoic acid, 6-aminohexanoic acid, 8-amino-3,6-dioxaoctanoic acid, 12-amino-4,7,10-trioxadodecanoic acid, 15-amino-4,7,10,13-tetraoxapentadecanoic acid and trioxatridecan-succinamic acid. Preferably, when the linker is connected to an amino acid lysine, the linker is selected from NHS-ester, isocyanate, benzoyl fluoride or carbamate.

[0530] According to the above different embodiments, the present invention relates to a compound comprising or consisting of an IL-17A binding peptide of formula (I) or (II) above, and having the following formula (IV):

[0531] (IV) [Biomolecule or cap] a -[Connector] b –(Sequence A) n -A m -[Peptide]–A' m’ -(Sequence A) n’ -[Connector] b’ –[Biomolecule or Cap] a’

[0532] Among them, independently of each other:

[0533] a=0 or 1;

[0534] b = 0 or 1;

[0535] a'=0 or 1;

[0536] b' = 0 or 1;

[0537] m = 0 to 10;

[0538] m' = 0 to 10;

[0539] n = 0 or 5;

[0540] n' = 0 or 5,

[0541] and

[0542] The peptide is a polypeptide according to formula (I) or formula (II);

[0543] A or A' is an amino acid selected from R, K, G, E or A, repeated m or m' times;

[0544] and sequence A comprises the sequence of formula (III) above;

[0545] The biomolecule is independently hexanoic acid, ascorbic acid, NAG-NAM, NAG, NAM, hyaluronic acid, alginic acid, chitin, (GalNAc)2, Gal-α1,3-GalNAc or trigalacturonic acid;

[0546] The caps are independently amide, aldehyde, ester, p-nitroaniline, 7-amino-4-methylcoumarin, acetyl, formyl, pyroglutamyl or fatty acid;

[0547] The linkers are independently 4-aminobutyric acid, β-alanine, 2-aminoethoxy-acetic acid, 5-aminopentanoic acid, 6-aminohexanoic acid, 8-amino-3,6-dioxaoctanoic acid, 12-amino-4,7,10-trioxadodecanoic acid, 15-amino-4,7,10,13-tetraoxapentadecanoic acid or trioxatridecanoic acid.

[0548] According to a preferred embodiment, the sequence A is linked to the N-terminus or the C-terminus of the peptide of formula (I) or formula (II).

[0549] Some examples of possible compounds according to the invention are reported below, but all other possible combinations contained in formula (IV) may exist:

[0550] -[Biomolecule or Cap]-[Linker]-(Sequence A)-A-[Peptide]-;

[0551] -[biomolecule or cap]-(sequence A)-A-[peptide]-;

[0552] -[Biomolecule or Cap]--A-[Peptide]-;

[0553] -[Biomolecule or Cap]-[Peptide]-;

[0554] --[Peptide]-A m’ -(Sequence A) n’–[Biomolecule or Cap] a’ ;

[0555] --[Peptide]-A m –[Biomolecule or Cap] a’ ;

[0556] --[Peptide]–[Biomolecule or Cap] a’ ;

[0557] According to a preferred embodiment, the biomolecule in formula (IV) of the present invention is selected from ascorbic acid, hexanoic acid, NAG or NAM.

[0558] According to another preferred embodiment, the cap according to the invention is a C-terminal modification selected from amides, preferably N-alkylamides, aldehydes, esters, preferably methyl and ethyl esters, p-nitroaniline, 7-amino-4-methylcoumarin or an N-terminal modification selected from acetyl, formyl, pyroglutamyl, fatty acids, preferably hexanoic acid, urea, carbamates, sulfonamides or alkylamines, preferably, the cap is selected from amides, fatty acids such as hexanoic acid and acetyl.

[0559] According to the present invention, the linker, biomolecule or cap in formula (IV) may be identical to or different from each other.

[0560] In another preferred embodiment of the present invention, the linker of formula (IV) is selected from 4-aminobutyric acid, β-alanine, 2-aminoethoxy-acetic acid, 5-aminopentanoic acid or trioxatridecane-succinamic acid.

[0561] Preferably, when the linker is attached to an amino acid lysine, the linker is selected from NHS-ester, isocyanate, benzoyl fluoride or carbamate.

[0562] A seventh object of the present invention is a pharmaceutical composition comprising said peptide, dimer or bioconjugate according to the objects of the present invention as described above, and at least one pharmaceutically acceptable excipient.

[0563] The pharmaceutical compositions of the present invention may be formulated in a form suitable for topical or ophthalmic administration.

[0564] Preferably, when the pharmaceutical composition of the present invention is administered by a topical route, the pharmaceutical form is selected from creams, ointments, gels, salve, solutions, lotions, suspensions, drops, buffers (buffered solutions), suspensions, eye drops, drops, sprays, wipes or powders, preferably it is selected from creams, gels, sprays or ointments.

[0565] In ophthalmic administration, the pharmaceutical form is preferably selected from eye drops, eye gels, ointments, lotions, wipes, sprays or creams.

[0566] According to a specific embodiment, the peptide or bioconjugate of the invention is topically administered by using microparticles or nanoparticles.

[0567] According to the present invention, the pharmaceutical composition of the present invention can be administered to animals and humans, humans being defined as adults and the "pediatric population", wherein the term "pediatric population" denotes the population portion from birth to eighteen years of age.

[0568] An eighth object of the present invention is the above peptide, dimer or bioconjugate according to the present invention for use in the treatment and / or prevention of inflammatory diseases and autoimmune diseases.

[0569] A ninth object of the present invention is a method for treating and / or preventing inflammatory diseases and autoimmune diseases, which comprises administering a therapeutically effective amount of the above-mentioned peptide, dimer or bioconjugate according to the present invention to a patient in need thereof.

[0570] Preferably, the inflammatory disease and autoimmune disease are selected from rheumatoid arthritis, multiple sclerosis, Crohn's disease, systemic lupus erythematosus, asthma, Behcet's disease, high IgE syndrome, ankylosing spondylitis, psoriasis, psoriatic arthritis, rheumatoid arthritis, keratoconjunctivitis sicca, vernal keratoconjunctivitis, stromal herpetic keratitis, corneal allograft rejection, corneal infection, preferably herpes virus and Pseudomonas aeruginosa keratitis, and dry eye. More preferably, the disease is an autoimmune eye disease or skin disease, even more preferably, it is dry eye or psoriasis.

[0571] What all of the above named diseases and medical conditions have in common is that their origin and / or symptoms are associated with IL-17A and / or Th-17.

[0572] For the latter, more specifically, dry eye disease (DED) is a highly prevalent condition that includes a large number of ocular surface disorders, with ocular mucosal inflammation being a characteristic feature, which, if uncontrolled, may lead to vision loss due to inflammation-induced corneal ulcers and scarring. In the development of DED, pathogenic immune cells (mainly Th17 cells) continuously migrate to the surface of the ocular mucosa and secrete proinflammatory mediators including IL17, thereby causing ocular surface inflammation and epitheliopathy.

[0573] The peptides, dimers or bioconjugates of the invention are conveniently and preferably administered topically in the form of eye drops or ophthalmic gels and ointments.

[0574] In another embodiment, the peptides, dimers or bioconjugates used according to the invention are administered as the sole active ingredient, or in combination with other active ingredients and / or in combination with medical devices for the symptomatic treatment of ophthalmic conditions, including but not limited to DED, such as ocular lubricants or "artificial tears", topical re-epithelizing agents, therapeutic contact lenses and punctum plugs.

[0575] Preferably, the other active ingredient is an adjuvant, an immunosuppressant, an immunomodulator or an anti-inflammatory agent.

[0576] For example, the IL-17A-binding peptides of the present invention can be used in combination with DMSO.

[0577] According to a preferred embodiment, the IL-17A binding peptide of the present invention can be used in combination with: an immunosuppressive monoclonal antibody, such as a monoclonal antibody having affinity for a leukocyte receptor selected from MHC, CD2, CD3, CD4, CD7, CD8, CD25, CD28, CD40, CD45, CD58, CD80, CD86 or its ligands; other immunomodulatory compounds, preferably recombinant binding molecules having at least a portion of the extracellular domain of CTLA4 or a mutant thereof, at least an extracellular portion of CTLA4 linked to a non-CTLA4 protein sequence (e.g., CTLA4Ig, designated as ATCC 68629) or a mutant thereof, such as LEA29Y, or a mutant thereof; an adhesion molecule inhibitor, an LFA-I antagonist, an ICAM-1 or ICAM-3 antagonist, a VCAM-4 antagonist or a VLA-4 antagonist.

[0578] In another preferred embodiment, the IL-17A binding peptide of the present invention is used in combination with the following substances: DMARD (disease modifying antirheumatic drug), preferably gold salts, sulfasalazine, antimalarial, methotrexate, D-penicillamine, azathioprine, mycophenolic acid, cyclosporine A, tacrolimus, sirolimus, minocycline, leflunomide, glucocorticoids; calcineurin inhibitors, preferably cyclosporine A or FK506; lymphocyte recirculation regulators, preferably FTY720 and FTY720 analogs; m TOR inhibitors, preferably rapamycin, 40-O-(2-hydroxyethyl)-rapamycin, CCI779, ABT578, AP23573 or TAFA-93; ascomycins with immunosuppressive properties, preferably ABT-281, ASM981; corticosteroids; cyclophosphamide; azathioprine; methotrexate; leflunomide; mizoribine; mycophenolic acid; mycophenolate mofetil mofetil); 15-deoxyspergualine or its immunosuppressive homologues, analogs or derivatives; or a chemotherapeutic agent, preferably paclitaxel, gemcitabine, cisplatin, doxorubicin or 5-fluorouracil; an anti-TNF agent, preferably a monoclonal antibody against TNF, preferably infliximab, adalimumab, CDP870, or a receptor construct against TNF-RI or TNF-RII, preferably Etanercept, PEG-TNF-RI; a blocker of proinflammatory cytokines, an IL-1 blocker, preferably Anakinra or IL-1 trap, AAL160, ACZ 885, IL-6 blocker; inhibitor or activator of protease, preferably inhibitor or activator of metalloprotease, anti-IL-15 antibody, anti-IL-6 antibody, anti-IL-23 antibody, anti-IL-22 antibody, anti-IL-21 antibody, anti-IL-12 antibody, anti-IFN-γ antibody, anti-IFN-α antibody, anti-CD20 antibody, anti-inflammatory agent, preferably aspirin or anti-infective agent.

[0579] Naturally, this list of agents for co-administration is not limiting, nor complete.

[0580] Another object of the present invention is one of the peptides listed in Table 6 for use in the treatment and / or prevention of the above-mentioned inflammatory diseases and autoimmune diseases.

[0581] Table 6

[0582]

[0583]

[0584] The amount and mode of administration of a peptide according to the invention will vary according to the particular peptide inhibitor of the invention, the individual patient group or patient, the presence of other medically active compounds and the nature and severity of the condition being treated.

[0585] According to a preferred embodiment, the dosage for preventive and / or therapeutic use is about 5-50 μg / ml, preferably about 10-25 μg / ml.

[0586] Preferably, the frequency of administration for preventive and / or therapeutic use is in the range of about once to twice a day, preferably once a day.

[0587] The present invention is further described by way of illustration in the following examples, none of which should be construed as limiting the scope of the invention as outlined in the appended claims. Example

[0588] Example 1 Peptide synthesis and purification:

[0589] We have developed a homology model of IL-17A using available crystallographic data (PDB 5h13, PDB 5vb9, and PDB 4hsa) (Liu et al. Nat Commun 2013, 4, 1888, Liu et al. Sci Rep 2016, 6: 30859, Liu et al. Sci Rep 2016, 6, 26071, Ting, Tung et al. PLoS One 2018, 13 (1): e01908502018). In our model, we also reconstructed the unresolved loops in the crystal and optimized the model by molecular dynamics simulations performed using Desmond software, as implemented in the Schrodinger Maestro macromodel suite. All molecular simulations were run for 1 microsecond to ensure system stability.

[0590] Based on the structure and properties of the IL-17A alpha pocket, different peptide sequences have been designed to obtain i) stable secondary and tertiary conformations that bind IL-17A with optimized protein binding site occupancy and ii) physicochemical properties suitable for topical use in ophthalmic and dermatological pathologies (Liu et al. Nat Commun 2013, 4, 1888, Espada et al J Med Chem 2016, 59(5), 2255-2260).

[0591] Furthermore, we optimized the ligand-IL-17A interaction by introducing hydrogen bonds and optimizing hydrophobic interactions.

[0592] a. Fmoc-based solid phase peptide synthesis

[0593] All chemicals were purchased and used without further purification.

[0594] The peptides of SEQ ID N. 1 to 43 and 111 to 257 of Table 1 were prepared by manual or automated solid phase peptide synthesis on Rink Amide-MBHA resin as solid support and with an acetyl-capped N-terminal amine as described below.

[0595] The Fmoc (9-fluorenylmethoxycarbonyl) group for Nα-protection was cleaved by treatment with 20% piperidine in dimethylformamide (DMF) for 8 min, followed by further treatment with the same reagent for 10 min. After Fmoc cleavage, the peptide resin complex was washed with DMF (×6). The next residues were then incorporated by using 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium3-oxid hexafluorophosphate (HATU / N,N-diisopropylamine (DIPEA) coupling protocol [Fmoc-amino acid (3 equiv.), HATU (3 equiv.) and DIPEA (6 equiv.)]. After slow stirring (1 h) and washing with DMF (× 6), the peptide resin was analyzed by Kaiser test. After assembly, the peptide resin was washed with DMF (× 3), dichloromethane (DCM) (× 4) and then dried in vacuo.

[0596] Cleavage and deprotection

[0597] The peptide was cleaved from the resin and, if necessary, deprotected by using a mixture of trifluoroacetic acid / 1,2-ethanedithiol / thioanisole / phenol / H2O / triisopropylsilane in a volume ratio of 68.5 / 10 / 10 / 5 / 3.5 / 1 as a cleavage solution. 3 ml of cleavage solution was used per 100 mg of resin. Complete deprotection was achieved by stirring at 30°C for 4 hours. After the cleavage reaction, the peptide in the mixture was precipitated in cold ether and dried in vacuo at 50°C.

[0598] b. Disulfide bond formation in peptide 41

[0599] The disulfide bond formation in the peptide of SEQ ID N.41 of Table 1 was obtained by incubating the peptide in a DMSO / H2O (20 / 80, v / v) mixture, which was monitored by RP-HPLC, MS and free thiol detection (DTNB method).

[0600] c. Purification by preparative RP-HPLC and purity assessment by analytical HPLC

[0601] A C18 column (10 μm, The crude peptide sample was purified by preparative RP-HPLC (Agilent) using a solvent system consisting of solvent A (0.1% TFA, 2% CH3CN in water) and solvent B (90% CH3CN / H2O), eluted at a flow rate of 25 mL / min, and detected absorbance at 220 nm. The solvent was removed by lyophilization. The final product was characterized by MALDI-TOF-MS, and the purity of the material was evaluated by analytical RP-HPLC (C18-250 mm×4.6 mm ID, flow rate of 1 mL / min), and the absorbance was detected at 220 nm.

[0602] Embodiment 2: Preparation of NAM and / or NAG bioconjugated peptides BIO-1, BIO-2 and BIO-3

[0603] Bioconjugated peptides BIO-1, BIO-2 and BIO-3 were prepared by conjugation reaction of peptide 42 or 43 according to the following procedure.

[0604] 1. Preparation of donor and acceptor

[0605] All reactions were carried out under N2 atmosphere. NMR spectra were run on a Brucker 400MHz instrument. HPLC-UV analysis was obtained on an Agilent 1260 Infinity system equipped with a G1311B Agilent 1260 quaternary pump, a G1329B Agilent 1260 autosampler, a G1315C Agilent 1260 diode array detector and a G1316A Agilent 1260 column thermostat module. Phenomenex GEMINI C18 150×4.6mm2 (5 μm) column was used. Mobile phase A was MilliQ water with 0.05% TFA, and mobile phase B was HPLC grade acetonitrile with 0.05% TFA, with a flow rate of 1.0 mL / min. The HPLC system was connected to an Agilent Quadrupole 6120 LC / MS mass spectrometer running in positive ion mode. Electrospray ionization source was used to generate ions. The acquired data were processed using Agilent Chemstation software.

[0606] a. Synthesis of 1-(2,2,2-trichloroacetimido)2-deoxy-3-O-acetyl-4,6-O-benzylidene-2-(2,2,2-trichloroethoxycarbonylamino)-aD-pyranoglucoside (donor)

[0607] At room temperature, trichloroethoxycarbonyl chloride (2.1 mL, 15.3 mmol) was added dropwise to a vigorously stirred solution of D-glucosamine hydrochloride (3 g, 13.9 mmol) and NaHCO3 (3.5 g, 41.7 mmol) in water (30 mL). The mixture was stirred for 2 h, then neutralized with 1 M HCl, concentrated, and dried in vacuo. 2-deoxy-2-(2,2,2-trichloroethoxycarbonylamino)-aD-pyranoglucoside (Intermediate A) was obtained.

[0608] The crude product was used directly in the next reaction step without further purification.

[0609] Specifically, zinc chloride (1.9 g, 13.9 mmol) was added to a solution of intermediate A (4.9 g, 13.9 mmol) in benzaldehyde (24 ml) and molecular sieves. ( 600mg) in a solution. After stirring at room temperature overnight, saturated NaHCO3 aqueous solution (30mL) and ether (90mL) were added, and the reaction mixture was stirred for 15min. The formed precipitate was filtered, washed with water, ether and dried. The residue was dissolved in pyridine (13mL) cooled to 0°C, treated with acetic anhydride (6.4mL, 68mmol), and stirred at room temperature overnight, then the solution was concentrated with toluene, cooled to 0°C, and extracted with DCM and saturated NaHCO3 aqueous solution (3x), then washed with saturated NaCl solution. The combined organic layer was dried, concentrated, and the crude product was purified by Isolera (EtPet / EtOAc) to obtain 1,3-di-O-acetyl 2-deoxy-4,6-O-benzylidene-2-(2,2,2-trichloroethoxycarbonylamino)-aD-pyranoglucoside (Intermediate B) as a white solid (3.05g, 41.7% yield).

[0610] Then morpholine (1.2mL, 13.9mmol) is added in anhydrous ethyl acetate (EtOAc) (12.2mL) solution of intermediate B (3.05g, 5.8mmol).After stirring at room temperature overnight, the reaction mixture is quenched with 3N HCl solution (3.5mL), then stirred for 20min.Extracted with EtOAc, and washed with water, saturated NaHCO3 aqueous solution and saturated NaCl solution, dried and concentrated.By Isolera (EtPet / EtOAc) purification of crude product, to obtain 2-deoxy-3-O-acetyl-4,6-O-benzylidene-2-(2,2,2-trichloroethoxycarbonylamino)-aD-pyranoglucoside (intermediate C), it is white solid (2.2g, 78.4%). To anhydrous DCM (44 ml) solution of intermediate C (2.2 g, 4.54 mmol) was added CsCO (680 mg, 2.1 mmol) and CClCN (4.4 mL, 43.8 mmol). The reaction mixture was stirred at room temperature for 2 h, then filtered through diatomaceous earth and concentrated to give 1-(2,2,2-trichloroacetimido)2-deoxy-3-O-acetyl-4,6-O-benzylidene-2-(2,2,2-trichloroethoxycarbonylamino)-aD-pyranoglucoside (donor) as a light yellow solid (2.2 g, 77%).

[0611] b. Synthesis of 1,6-di-O-benzyl 2-deoxy-3-O-((R)-10-ethoxycarbonylethyl)-2-(2,2,2-trichloroethoxycarbonylamino)-aD-pyranoglucoside (acceptor)

[0612] At room temperature, allyl chloroformate (6.6 mL, 62.2 mmol) was added dropwise to a vigorously stirred solution of D-glucosamine hydrochloride (12.2 g, 56.6 mmol) and NaHCO3 (14.3 g, 169.7 mmol) in water (61 mL). The mixture was stirred for 2 h, then neutralized with 1 M HCl, concentrated, and dried in vacuo. The crude product 2-deoxy-2-(allyloxycarbonylamino)-aD-pyranoglucoside (intermediate D) was obtained. The product was used in the next reaction step without further purification. Intermediate D (14.9 g, 56.6 mmol) was dissolved in benzyl alcohol (89 mL), and acetyl chloride (15.4 mL, 215.2 mmol) was added dropwise at 0 ° C. After stirring at 80 ° C for 3 h, the reaction mixture was quenched with a cold saturated NaHCO3 aqueous solution (20 ml) and stirred for 30 min. Cold water and ether were added, and the reaction mixture was stirred for 30 min. The two phases were separated, and the aqueous phase was concentrated and dried in vacuo. 30 ml of water and 200 ml of ether were added until a precipitate was formed, which was filtered and washed several times with cold ether (until no trace of benzyl alcohol was detected). Benzyl 2-deoxy-2-(allyloxycarbonylamino)-aD-pyranoglucoside (Intermediate E) was obtained as a white solid (11 g, 55%) and was used directly in the next reaction step without further purification. Zinc chloride (4.2 g, 31.1 mmol) was added to a solution of benzaldehyde (55 ml) and intermediate E (11 g, 31.1 mmol). MS (4.9g) solution. After 2h, zinc chloride (4.2g, 31.1mmol) was added. After stirring overnight at room temperature, the reaction mixture was treated with saturated NaHCO3 aqueous solution (70mL), petroleum ether (420mL), and stirred for 10min. The precipitate was filtered, washed with petroleum ether and dissolved in DCM. The organic solution was extracted with saturated NaHCO3 aqueous solution, water and saturated NaCl solution, dried and concentrated. The crude product was purified by Isolera (DCM / EtOAc) to obtain benzyl 2-deoxy-4,6-O-benzylidene-2-(allyloxycarbonylamino)-aD-pyranoglucoside (intermediate F) as a white solid (2.75g, 20%).

[0613] Sodium hydride (NaH, 348.8 mg, 8.7 mmol, 60% oil dispersion) was added to a solution of intermediate F (2.75 g, 6.2 mmol) in anhydrous DCM (40.6 mL) cooled to 0 ° C, and the mixture was stirred at room temperature for 30 min. The mixture was then treated with pure (-)-ethyl (S)-2-trifluoromethanesulfonic acid propyl ester and stirred at room temperature for 2 h. The reaction mixture was quenched by adding ice and extracted with DCM. The organic solution was washed with saturated NaHCO3 aqueous solution, saturated NaCl solution, dried and concentrated. The residue was purified by Isolera (DCM / EtOAc) to obtain benzyl 2-deoxy-3-O-((R)-10-ethoxycarbonylethyl)-4,6-O-benzylidene-2-(allyloxycarbonylamino)-aD-pyranoglucoside (intermediate G) as a white solid (2.35 g, 69.7%).

[0614] Tetrakis (triphenylphosphine) palladium (1.5g, 1.3mmol) and acetic acid (AcOH, 0.385mL, 6.725mmol) are added to anhydrous DCM (42.7mL) solution of intermediate G (2.35g, 4.34mmol). The reaction mixture is stirred at room temperature for 15min, then chloroformic acid 2,2,2-trichloroethyl ester (TrocCl, 1.224mL, 8.895mmol) is added dropwise, and the resulting solution is stirred at room temperature for 1h. The reaction mixture is quenched with saturated NaHCO3 aqueous solution, extracted with DCM, washed with H2O and saturated NaCl solution. After vacuum concentration, the residue is dissolved in ether (100ml), and insoluble matter is filtered out. The organic phase was dried, concentrated and purified by Isolera (cyclohexane / AcOEt) to give benzyl 2-deoxy-3-O-((R)-10-ethoxycarbonylethyl)-4,6-O-benzylidene-2-(2,2,2-trichloroethoxycarbonylamino)-aD-glucopyranoside (Intermediate H) as a white solid (2.3 g, 83.7%).

[0615] To a solution of intermediate H (2.4 g, 3.8 mmol) in anhydrous CHCN (38 mL) was added a solution of MeN-BH (332 mg, 4.55 mmol) in CHCN (2.2 mL) at 0° C., followed by a solution of BF-OEt (2.89 mL, 23.4 mmol) in CHCN (8 mL) dropwise. After stirring at 0° C. for 3 h, the mixture was quenched with cold saturated aqueous NaHCO (30 mL), diluted with EtOAc (350 ml), and washed with saturated aqueous NaHCO (100 ml), 5% citric acid (4×50 ml), saturated aqueous NaHCO and saturated NaCl solution (40 mL). The organic layer was dried, concentrated, and the crude product was purified by Isolera (cyclohexane / AcOEt) to give 1,6-di-O-benzyl 2-deoxy-3-O-((R)-10-ethoxycarbonylethyl)-2-(2,2,2-trichloroethoxycarbonylamino)-aD-glucopyranoside (acceptor) as a colorless foam (1.2 g, 49.8%).

[0616] 2. Synthesis of NAG-NAM conjugated peptide BIO-3

[0617] Will be suspended in The donor (1.6 g, 2.6 mmol) and the acceptor (1.1 g, 1.73 mmol) in anhydrous DCM (55 mL) of molecular sieves (200 mg) were treated with trimethylsilyl trifluoromethanesulfonate (TMSOTf, 188 μL, 1.04 mmol) at -15 ° C. After stirring for 20 min, 0.75 equivalents of donor and 0.3 equivalents of TMSOTf were added. After stirring for 20 min, 0.75 equivalents of donor and 0.3 equivalents of TMSOTf were added. The mixture was then quenched with a cold saturated NaHCO3 aqueous solution (15 mL) and extracted with DCM (60 mL). The organic layer was washed with a saturated NaHCO3 aqueous solution and a saturated NaCl solution, dried and concentrated. The residue was purified by Isolera reverse phase (H2O / ACN neutral phase) to give benzyl 6-O-benzyl-4-O-[2-deoxy-3-O-acetyl-4,6-O-benzylidene-2-(2,2,2-trichloroethoxycarbonylamino)-bD-glucopyranosyl]-2-deoxy-3-O-[(R)-1'-ethoxycarbonylethyl]-2-(2,2,2-trichloroethoxycarbonylamino)-aD-glucopyranoside (Intermediate L) as a white foam (1.2 g, 62.9%).

[0618] A solution of intermediate L (1.2 g, 1.09 mmol) and zinc–copper couple (3.12 g, 24.2 mmol) in a mixture of AcOH / Ac2O / THF 1:1:1 (12 mL) was stirred at room temperature for 4 h. The reaction mixture was filtered through celite, washed with EtOAc and concentrated. The crude product was concentrated and purified by column chromatography (cyclohexane / AcOEt) to give benzyl 6-O-benzyl-4-O-[2-deoxy-3-O-acetyl-4,6-O-benzylidene-2-acetylamino-bD-pyranoglucopyranosyl]-2-deoxy-3-O[(R)-10-ethoxycarbonylethyl]-2-acetylamino-aD-pyranoglucopyranoside (Intermediate M) as a white solid (560 mg, 0.67 mmol, 61.6%). HPLC-MS (ESI+) m / z: C 44 H 54 N2O 14 +Na:857.3472. Measured:857.6124.

[0619] To a solution of intermediate M (91 mg, 0.109 mmol) in THF / 1,4-dioxane / H2O 4:2:1 (2.8 mL) was added LiOH.H2O (56 mg, 1.34 mmol). After stirring at room temperature for 2 h, the reaction mixture was filtered through Dowex H+ (freshly activated with 1N HCl). The residue was purified by chromatography on a diaion HP-20 column (2×7 cm) previously washed with water, MeOH, and water. The column was eluted first with H2O (50 mL) and then with MeOH (30 mL). The alcohol fraction was concentrated to give benzyl 6-O-benzyl-4-O-[2-deoxy-4,6-O-benzylidene-2-acetylamino-bD-glucopyranosyl]-2-deoxy-3-O-[(R)-10-ethoxycarbonylethyl]-2-acetylamino-aD-glucopyranoside (Intermediate N) as a whitish solid (57.4 mg, 69%). Mp 102–105℃,1HNMR(600MHz,CD3OD)δH 7.51–7.27(m,15H,ArH),5.59(s,1H,CHPh),5.36(d,J 3.1Hz,H-1),4.85(m)H2O,H-10,1 / 2 CH2Ph),4.66–4.57(m,3H,J 12.1Hz,CHCH3,CH2-Ph),4.46(d,1H,J 12.2Hz,1 / 2 CH2Ph),4.29(dd,1H,J 10.3Hz,J 5.0Hz,H-60b),4.07(t,1H,J 9.6Hz,H-30),3.95(t,1H,J 9.1Hz,H-4),3.82–3.77(m,2H,H-6b,H-3),3.72–3.62(m,3H,H-60a,H-5,H-6a),3.55–3.40(m,3 H,H-20,H-40,H-2),3.30–3.27(m,1H,H-50),1.98(s,3H,COCH3),1.96(s,3H,COCH3),1.37(d,J 6.9Hz,3H,CHCH3). HPLC-MS(ESI+)m / z: C 40 H 48 N2O 13 +Na:787.3054. Measured:787.7830.

[0620] The peptide of SEQ ID N.42 was coupled to compound N under the conditions used for amino acid coupling, and the resulting bioconjugate was cleaved from the solid support by treatment with trifluoroacetic acid (1% in DCM). The resulting bioconjugate (0.026 mmol) was dissolved in acetic acid (8 mL) and Pd(OH)2 / C (58 mg) was added. The resulting mixture was stirred at room temperature for 6 h under a hydrogen atmosphere. The mixture was filtered through celite and concentrated to give the bioconjugate BIO-3 in quantitative yield.

[0621] The bioconjugate was cleaved from the resin and the last residue was deprotected using a mixture of trifluoroacetic acid / 1,2-ethanedithiol / thioanisole / phenol / H2O / triisopropylsilane in a volume ratio of 68.5 / 10 / 10 / 5 / 3.5 / 1 as the cleavage solution. 3 ml of cleavage solution was used per 100 mg of resin. Complete deprotection was achieved after 4 h at 30 °C. After the cleavage reaction, the crude peptide was precipitated by adding cold ether and dried in vacuo at 50 °C.

[0622] A C18 column (10 μm, The crude bioconjugate was purified by preparative RP-HPLC (Agilent) at 50 × 250 mm (50 × 250 mm). A solvent mixture consisting of solvent A (0.1% TFA, 2% CH3CN in H2O) and solvent B (90% CH3CN / H2O) was used for elution at a flow rate of 25 mL / min, and the absorbance was detected at 220 nm. The solvent was removed by lyophilization, and the final product was characterized by MALDI-TOF-MS. The purity of the purified product was evaluated by analytical RP-HPLC (C18-250 mm × 4.6 mm ID, flow rate of 1 mL / min), and the absorbance was detected at 220 nm.

[0623] 3. Synthesis of NAG and NAM bioconjugates BIO-1 and BIO-2

[0624] The synthesis of NAM-peptide and peptide-NAG was carried out starting from the peptide-anchored resin (Example 1), wherein the peptide was the peptide of SEQ ID N.42 (for BIO-1) or the peptide of SEQ ID N.43 (for BIO-2), respectively, and the donor and acceptor were the molecules described above according to the procedure in Swaminathan et al., Proc Natl Acad Sci USA. 2006, 17; 103(3): 684-9.

[0625] Example 3: Synthesis of Decanoic Acid Bioconjugate BIO-4

[0626] The decanoic acid-capped peptide was synthesized according to the procedure described in Example 1 by using a decanoic acid-capped N-terminal amine.

[0627] Example 4: Synthesis of Ascorbic Acid Bioconjugate BIO-5

[0628] 5,6-O-isopropylidene-L-ascorbic acid (iASA) was synthesized by the method of Jung (Jung, ME; Shaw, TJJAm. Chem. Soc. 1980, 102, 6304). For activation, CDI (10 equivalents) was added to the peptide-anchored resin for 2 h. The activation of the N-terminal amine of the peptide after cleavage from the resin was confirmed by RP-HPLC using the following conditions: A to B (A: 0.1% TFA in H2O, B: 0.1% TFA in CH3CN; 0% to 60% B in 30 min, flow rate of 1.0 mL / min); UV 230 nm was detected. Next, iASA was introduced into the activated peptide-anchored resin. The product was separated from the resin by treatment with 50% TFA / DCM (v / v) for 1 h. The resin was filtered, the filtrate was concentrated under high vacuum, and precipitated with cold ether. The structure of the resulting bioconjugate BIO-5 was confirmed by LC / MS at a flow rate of 250 μL / min for 30 min using 0.1% formic acid / methanol as eluent and monitoring at 230 nm.

[0629] Example 5: IL-17RA / IL17RC dimerization assay in response to IL-17A

[0630] The peptides of Table 7 were tested for their ability to inhibit the binding of IL-17A to its receptor and the subsequent interaction of IL-17RA and IL17RC.

[0631] The interaction of the receptor chains IL-17RA and IL17RC after activation by IL-17A was measured in the absence or presence of the peptides of Table 7 using the interleukin receptor RA and RC dimerization assay (DiscoverX).

[0632] exist In the cytokine receptor assay, one cytokine receptor chain is labeled with a small peptide epitope, ProLink (PK), and the other chain is labeled with an enzyme receptor (EA). Ligand binding induces dimerization of the two receptors, promoting complementation of the PK and EA fragments. This interaction produces units of b-galactosidase activity, which are detected using a chemiluminescent substrate. The PathHunter cell line is amplified from frozen stocks according to standard procedures. Cells are seeded into white-walled 384-well microplates in a total volume of 20 μL and incubated for the appropriate time before testing. To inhibit agonist activity, cells are incubated with samples to induce a response. Intermediate dilutions of sample stocks are made to generate 5X samples in assay buffer. 5 μL of 5X sample is added to the cells and incubated for 60 minutes at 37°C or room temperature. The vehicle concentration is 1%. 5 μL of 6X EC in assay buffer is added. 80Agonist (IL-17A) was added to the cells and incubated at 37°C for 6 to 16 hours depending on the assay. 1. The assay signal was generated by a single addition of 12.5 μL or 15 μL (50% v / v) of the PathHunter detection reagent mixture for the agonist (IL-17A) and (peptide of Table 7) assays, respectively, followed by incubation at room temperature for one hour. For some assays, activity was detected using a high-sensitivity detection reagent (PathHunter Flash Kit) to improve assay performance. In these assays, an equal volume of detection reagent (25 μL or 30 μL) was added to the wells and then incubated at room temperature for one hour. After the signal was generated using a PerkinElmer EnvisionTM instrument for chemiluminescent signal detection, the microplate was read. The activity of the compound was analyzed using the CBIS data analysis suite (ChemInnovation, CA). Inhibition % = 100% x (1-(average RLU of test sample-average RLU of vehicle control) / (average RLU of EC80 control-average RLU of vehicle control)).

[0633] As reported in Table 7 below, all tested molecules were found to have activity in inhibiting IL-17RA / IL17RC dimerization, and therefore IL-17A pathway activation, showing IC50 values ​​in the mid nanomolar range.

[0634] Table 7

[0635]

[0636]

[0637]

[0638]

[0639]

[0640]

[0641] exist Figure 1 In, dose-response curves for exemplary peptides are reported.

[0642] Example 6: Phenotypic and viability assays of human differentiated TH17 cells

[0643] In order to evaluate the effects of the peptides of the invention on target cells causing inflammation in ophthalmic or dermatological pathologies, we tested the ability of the peptide of SEQ ID N. 18 to inhibit the secretion of proinflammatory cytokines and metalloproteinases by human Th17 cells in vitro.

[0644] Human primary CD4 T cells were differentiated into Th17 cells in vitro by culturing the cells in Th17 differentiation medium for 10 days. In detail, purified human peripheral blood CD4 T cells were cultured in Th-17 differentiation medium (CellXVivo Human Th17, Tocris) according to the manufacturer's instructions. In brief: 96-well tissue culture plates were coated with anti-CD3 and anti-CD28 agonistic antibodies. Cells were counted by trypan blue exclusion and suspended in Th-17 differentiation medium at a concentration of 1E5 cells / ml. 0.2 mL of this suspension was plated into each well of a 96-well plate. The cells were incubated at 37°C, 5% CO2 for 10 days, and the medium was changed as needed. After 10 days of cell differentiation, the peptides synthesized in Example 1, such as the peptide of Seq. ID No 18, were diluted to an appropriate concentration in CGM (cell growth medium, XVivo-15, Lonza). The cell plate was centrifuged at 300xg for 5 min, and the Th17 differentiation medium was removed.

[0645] After differentiation, cells were stimulated with an activation cocktail containing phorbol 12-myristate 13-acetate and ionomycin in the presence of varying concentrations of the test article.

[0646] 24h after stimulation, the expression of IL-17, IL-6, IL-23, MMP3 in the culture supernatant was analyzed, and the viability was checked using the fluorescent viability dye Alamar blue. In response to stimulation with the activation mixture, CD4 cells secreted high levels of IL-17A, indicating successful differentiation into Th17 cells. In addition, the stimulated cells secreted IL-6, IL23 and MMP3.

[0647] The supernatant was removed and replaced with 200 μL of diluted peptide or vehicle (CGM+DMSO diluted at TI concentration) in the appropriate group. Before stimulation, the cells were incubated with the peptide test for 1 hour. The following stimulation protocol was generated: 10x cell activation mixture: diluted from 500x to 10x working solution in CGM. 1 hour after adding TI, 22 μL of suitable stimulants were added to the wells. Finally, 25 μL of Alamar blue reagent (10x) was added to each well 20 hours after stimulation (4 hours before serum collection 24 hours) to determine cell viability. 24 hours after stimulation, Alamar blue fluorescence was measured according to the manufacturer's instructions. The remaining supernatant was collected for subsequent multiple protein analysis. The Luminex Multiplex platform on the MAGPIX instrument was used to evaluate the expression levels of IL-6, IL-17, IL-23 and MMP3 in the culture supernatant stimulated for 24 hours.

[0648] Figure 2 and Figure 3The effect of the peptide of SEQ ID N.18 is shown. It can be seen that the peptide can significantly reduce the secretion of IL-17A, IL-6 and IL-23 and the secretion of MMP3. Figure 4 As shown, the inhibition of cytokine and metalloproteinase secretion is not due to a problem with cell viability, since treatment of cells with the peptide of SEQ. ID N. 18 for 11 days did not impair cell viability.

[0649] Example 7: Physicochemical Analysis of Peptides

[0650] The physicochemical properties of the prior art peptide HAP and some peptides according to the invention (see Table 8) were tested as described below.

[0651] i. method

[0652] a. Isoelectric point IP logD and logP

[0653] The main physicochemical properties (isoelectric point (IP), logD (7.4) and logP) of the peptides in Table 8 below were determined using a Sirius T3 instrument (Sirius Analytical Instruments Ltd., East Sussex, UK) equipped with a Ag / AgCl double junction reference pH electrode, a Sirius D-PAS spectrometer and a turbidity sensing device. The pH electrode was calibrated with titrimetric analysis in the pH range of 1.8-12.2. An overhead stirrer was used, and a temperature probe (Peltier controller) was used to monitor the temperature during the titration. The titration experiments were carried out in a 0.15M KCl ionic strength adjustment solution (ISA water), a nitrogen atmosphere, and a temperature of 25±1°C. All tests were performed using standardized 0.5M KOH and 0.5M HCl as titrants, and for the partition coefficient test, saturated octanol in ISA water (5% ISA water) was used as the partition solvent. pKas were determined using pH titration by potentiometric method. The powder (approximately 0.5 mg) was dissolved in 1.5 ml ISA water and titrated in triplicate over the pH range of 2.0–12.0. Each LogP was determined in triplicate by dissolving the powder (approximately 1 mg) in 1 ml ISA water and then titrating at pH in three different percentages of octanol (typically 50%, 60%, 70%).

[0654] b. Water solubility and stability

[0655] General procedure for preparing stock solutions

[0656] Each lyophilized peptide was weighed into an amber vial. A calculated amount of phosphate buffer at pH 7.8 was added so as to obtain a 10 mM stock solution directly. The recovered suspension was stirred for 30 minutes at 37°C in an orbital shaking incubator. Subsequent dilutions were performed to obtain more dilute stock solutions of less soluble compounds.

[0657] To evaluate stability and solubility, all stock solutions were stored at 25° C. Stability and solubility were evaluated by UHPLC-MS analysis compared with standard solutions at 3 time points: T0, 3 days, 1 week.

[0658] Chromatograms were evaluated for quantitative analysis and MS spectra were examined to support stability studies.

[0659] General Procedure for Preparation of Standard Solutions

[0660] Each peptide was weighed and directly diluted to 100 μM with phosphate buffer, pH 7.8. The solution was vortexed for 5 minutes and heated at 37°C for 20 minutes. The obtained standard solution was stored at 25°C in the dark under controlled temperature.

[0661] General procedure for sample preparation

[0662] Part of the stock solution was collected in an amber vial and diluted to 100 μM with phosphate buffer, pH 7.8 (dilution factor 1:100).

[0663] Instrumental methods

[0664] Reverse phase UHPLC was performed using a Luna Omega Polar C18 column (pore size 1.6 μm, 2.1 x 100 mm) at a flow rate of 0.3 ml / min. Solvent A was 0.5% formic acid in water, and solvent B was 0.5% formic acid in acetonitrile. First, the sample was loaded onto the column and washed isocratically at 10% B for 3 min. Then, a gradient of 10-95% B was run in 22 min, held at 95% for 2 min, then dropped to 10% in 1 min and held for 5 min to keep the pressure stable. UV absorption was monitored by DAD (220–400 nm). TIC was monitored using an ESI-MS ion trap.

[0665] ii. result

[0666] All peptides tested showed stability at 25 °C for up to 1 week as no obvious changes were found in the chromatograms and mass spectra.

[0667] Table 8 below shows the physicochemical properties of the peptides tested.

[0668] Table 8

[0669]

[0670]

[0671] As can be seen from the above, most of the peptides according to the present invention have optimal physicochemical properties, showing a good balance between solubility and lipophilicity. The mean LogD of the peptides is -2.64 ± 2.06, and for most of the peptides, the solubility observed in water is very good (> 10mM). The observed properties make it particularly suitable for ophthalmic and dermatological applications, because both hydrophobic and hydrophilic components are present in the skin and eye surface. They also explain the good permeability observed by the peptides (see Example 8).

[0672] Embodiment 8: Penetration Testing

[0673] The peptides tested were those of SEQ ID N. 1, SEQ ID N. 2, SEQ ID N. 7, SEQ ID N. 11, SEQ ID N. 14, SEQ ID N. 38, SEQ ID N. 111, SEQ ID N. 113, SEQ ID N. 115, and SEQ ID N. 121. They were dissolved in cell culture grade water (Coming, Manassas, VA, USA) to obtain 10 mM stock solutions.

[0674] i. method

[0675] In vitro models

[0676] I-HCEC, an immortalized human corneal epithelial cell line, is derived from primary human corneal epithelial cells (purity>99%). These cells are suitable for studying health and disease of the human cornea. Cells were cultured according to the protocol and using the medium recommended by nnoprot, Bizkaia, Spain (IM-Corneal Epithelial Cell Medium, reference number P60131).

[0677] Immunofluorescence

[0678] Before drug testing, qualitative and quantitative techniques were developed to first confirm and quantify barrier integrity. First, we performed immunofluorescence analysis of a protein characteristic of tight junctions (Z0-1) (Sugrue SP et al., (1997) Exp Eye Res. Jan; 64 (1): 11-20). Cells were seeded at a density of 1*104 cells / cm2 on coverslips coated with 50 μg / ml collagen I (stock solution was 3 mg / ml, from Gibco, NY, USA) and cultured in vitro for 14 days (14 DIV), with the medium replaced every day after confluence. Cells were fixed in methanol at -20°C for 10 min, rinsed with phosphate buffered saline (PBS), incubated in PBS containing 4% bovine serum albumin (BSA; Sigma-Aldrich) for 10 min at room temperature (RT), and incubated with primary and secondary antibodies for 60 min at room temperature, respectively. The primary antibody was goat anti-human Z0-1 (1:100, Thermo Fisher) at 4°C overnight, followed by anti-goat Alexafluor 488 (1:10000). Finally, the cells were counterstained with DAPI (0.5 mg / ml) (Sigma-Aldrich) for 5 min at room temperature, mounted with Vectashield mounting medium (Vector Laboratories Inc, Burlingame, CA), and observed under a confocal laser scanning microscope (Leica TCS SP5, Wetzlar, Germany).

[0679] MTS assay

[0680] Cell viability was determined at 24 h using a Cell Titer One Solution Cell Proliferation Assay (Promega Corporation Madison, WI, USA), which is a colorimetric method based on 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS). The amount of formazan formed was measured at 492 nm using an ELISA plate reader Infinite F200 (Tecan, Mannedorf, Swiss) as a function of viability. The assay was performed at 50 μM and 100 μM (N=9). The results are expressed as absorbance at 492 nm.

[0681] TEER assay

[0682] The barrier integrity of corneal epithelial cultures was verified by measuring TEER (transepithelial / transendothelial electrical resistance) and the permeability of paracellular permeates (e.g., mannitol, 6-carboxyfluorescein) (Ronkko S et al., (2016) Drug Deliv Transl Res 6: 660-675). TEER is a widely accepted quantitative technique for measuring the integrity of tight junction dynamics in endothelial and epithelial monolayer cell culture models. The TEER value is an indicator of cell barrier integrity before evaluating drug or chemical transport. TEER measurements can be performed in real time without damaging cells, and are generally based on measuring permeation resistance or impedance over a wide frequency range (Snirivasan et al., 2015 J Lab Autom. 20 (2): 107-126). In order to perform the assay, the Millicell ERS-2 resistance system of EMD Millipore (Burlington, MA, USA) was used according to the manufacturer's instructions. The assay was performed at 50 μM and 100 μM (N = 9). The results are expressed as resistance (Q / cm2).

[0683] Penetration Testing

[0684] The aim of this study was to evaluate the apparent permeability coefficient (Papp) values ​​of peptides across stratified human corneal epithelial cells. All peptides were tested at 50 μM in duplicate.

[0685] In particular, human corneal epithelial cells (hCEPIC) were layered on Transwell cell culture inserts (Corning, NY, USA). Permeation experiments were performed at 5 minutes, 30 minutes, and 60 minutes from the receiving chamber sampling and at the same time point from the donor chamber sampling. Each aliquot collected was analyzed by an HPLC instrument (Knauer) equipped with a C18 Vertex Plus column (Knauer) thermostatically controlled at 40°C. The ratio (apparent permeability coefficient, Papp) of the research compound was calculated and compared with the Papp values ​​of low permeability markers and high permeability markers 6-carboxyfluorescein and rhodamine B, respectively.

[0686] The apparent permeability coefficient (Papp) is calculated according to the following formula:

[0687] Papp=(l / A*C0)(dM / dt)

[0688] where dM / dt is the flux across the cell layer (nanomoles / second);

[0689] A (cm2) is the exposed surface area of ​​the inserted membrane;

[0690] C0 is the initial drug concentration (micromolar) in the donor compartment at t=0 (Xiang et al., Drug Metab Dispos. 2009 May; 37(5): 992-8).

[0691] Statistical analysis

[0692] All groups (studied compounds and rhodamine) were compared to low permeability controls. All other experiments are presented as mean ± standard error and significance was calculated by Student's t-test versus control cells. Differences were considered statistically significant (p < 0.05) as determined by Student's t-test.

[0693] ii. result

[0694] MTS and TEER assay

[0695] Under our experimental conditions, the peptides considered did not significantly affect cell viability or TEER at the tested concentrations of 50 μM and 100 μM.

[0696] Penetration Testing

[0697] The results obtained for each peptide are shown in Table 9 below.

[0698] Table 9

[0699]

[0700] To verify that HCE cell cultures can be used to screen peptides, the permeability of the lipophilic transcellular marker rhodamine B was determined. As expected, rhodamine B showed high permeability to HCE cell cultures (51.26 ± 5.44 10 -6 Then, the permeability of 6-carboxyfluorescein was measured and showed to be consistent with the literature (0.61 ± 0.33 10 -6 cm / sec). The marker has very low lipophilicity and penetrates biological membranes only through the intercellular spaces.

[0701] The tested peptides showed very good permeability to immortalized human corneal epithelium compared to the high permeability standard Rhodamine B. Obviously, the HCE permeability model does not include the stroma or endothelium, but these layers are not the key barriers to corneal drug absorption. We found peptides with good permeability (SEQ ID N.2) and peptides with higher permeability compared to Rhodamine B (SEQ ID N.115, SEQ ID N.113 and SEQ ID N.111).

[0702] Example 9: Peptide-IL-17A affinity test

[0703] The affinity of the prior art peptide HAP (SEQ ID N. 1) and the peptide of SEQ ID N. 2 to mIL-17A was tested as follows.

[0704] a. Immobilization of mIL-17A

[0705] To immobilize mouse IL-17A protein, amine coupling chemistry was used. The dextran matrix was first activated on the sensor chip surface with a mixture of 1-ethyl-3-carbodiimide (EDC) and N-hydroxysuccinimide (NHS) to obtain reactive succinimidyl esters. IL17 at 10ug / ml was diluted in 10mM acetate buffer at pH=5.0, and the ligand was then passed over the surface to allow the ester to react spontaneously with the uncharged amino groups, in this way covalently linking the ligand to the dextran. Covalent immobilization of mIL17 resulted in 3000RU of ligand stably attached to the surface.

[0706] b. Characterization of peptide / IL17 interactions

[0707] The ability of peptides of SEQ ID N.1 (HAP) and SEQ ID N 2 to bind to immobilized mIL-17A was analyzed. These molecules were diluted in HBS-EP buffer at different concentrations (62.5 nM to 1000 nM) and injected onto the sensor chip for 4 minutes, followed by 5 minutes of dissociation. After each run, the sensor chip was regenerated by injecting 2 M NaCl. Based on the sensorgrams ( Figure 1 , Panel A) and using the BIAevaluation software, Scatchard analysis of the RU values ​​at equilibrium allowed the determination of the affinity constant ( Figure 6 The results obtained are shown in Table 10 below.

[0708] Table 10

[0709] Compound Kdea(nM) SEQ ID N.1 3700 SEQ ID N.2 130

[0710] As shown in Table 8, the affinity value of the peptide of SEQ ID N.2 binding to IL-17A is equal to 130 nM, while the affinity value of the peptide of SEQ ID N.1 binding to IL17-A is very low (3700 nM).

[0711] In view of the results obtained in Examples 8 and 9, modification of the isoleucine at position 1 in the prior art HAP peptides with valine, as in the peptide of SEQ ID N 2, resulted in significant differences in physicochemical properties, permeability and affinity for ligands.

[0712] We summarize the properties of these two peptides below:

[0713] Table 11

[0714]

[0715]

[0716] Table 11 shows comparative data between peptides SEQ ID N.1 and SEQ ID N.2.

[0717] We found large differences in logP and logD (6-log and 3-log differences, respectively): this explains the higher permeability observed with SEQ ID N.2 compared to SEQ ID N.1 (28.8 vs. 19.3). The Kd (nM) measured in a surface plasmon resonance (SPR) assay against mouse IL-17A showed that SEQ ID N.2 had a 28-fold higher affinity than SEQ ID N.1.

[0718] Furthermore, molecular dynamics simulation studies were performed, which showed that the replacement of isoleucine (HAP) with valine (SEQ ID N. 2) greatly stabilized the folded structure: SEQ ID N. 2 was predicted to be 80% more stable.

[0719] Example 10: In vitro assessment of inflammatory cytokine expression

[0720] The peptide HAP (SEQ ID N. 1) and the peptide of SEQ ID N. 2 were tested in vitro to evaluate their direct effects on the expression of inflammatory cytokines in the cornea.

[0721] Immortalized human corneal epithelial cell line (I-HCEC) (Innoprot, Bizkaia, Spain, reference number P10871) was tested. This is derived from primary human corneal epithelial cells (purity> 99%) and is suitable for studying health and disease of the human cornea. Cells were cultured according to the protocol and using the medium recommended by Innoprot (IM corneal epithelial cell medium, reference number P60131). The cell culture medium was supplemented with 5% fetal bovine serum (FBS).

[0722] Cells were seeded at 320,000 cells / well in 6-well plates and 15,000 cells / well in 96-well plates. When cells reached 80% confluence, they were treated with SEQ ID N.1 and SEQ ID N.2 compounds at a dose curve (0.01-500 μM) for 24 hours.

[0723] According to the manufacturer's instructions, total RNA was isolated from 6-well plates using Quick-Start RNeasy Mini Kit (Qiagen, catalog number 74104). Reverse transcription was performed using SuperScript Vilo (reference number 11755050, Life Technologies), and real-time PCR was performed using the TaqMan protocol (reference number 4444557, Applied Biosystem) suitable for the CFX96 real-time system. The experiment was performed three times, each in duplicate. The probes used from Thermo Fisher Scientific were: human CXCL8 (Hs00174103_m1), human IL6 (Hs00174131_m1), human TNFα (Hs99999043_m1). GAPDH (probe Hs02758991_g1) was used as a housekeeping gene.

[0724] IL-8, IL-6 and TNF-α expression in I-HCECs was evaluated after 24 hours of treatment with different concentrations of peptides of SEQ ID N.1 or SEQ ID N.2 (0.01 μM, 0.1 μM, 1 μM, 10 μM, 100 μM, 500 μM). Untreated I-HCECs were used as negative controls (vehicle).

[0725] The results showed that compound SEQ ID N.2 did not change the expression of any of the inflammatory cytokines measured ( Figure 8 A, 9A and 10A), while treatment with 500 μM of compound SEQ ID N.1 significantly induced the expression of two inflammatory cytokines, IL-6 and TNFα ( Fig. 9 B and 10B).

[0726] The data obtained demonstrated that the peptide of SEQ ID N.1 had direct pro-inflammatory activity and significantly induced HCEC to express IL-6 and TNF-α inflammatory cytokines, which increased by 10-fold and 8-fold, respectively. In contrast, the peptide of SEQ ID N.2 did not show any effect on the expression of cytokines.

[0727] In view of the above, it can be concluded that the peptide of SEQ ID N. 2 surprisingly shows significantly better tolerability and lack of toxicity compared to the peptide HAP of SEQ ID N. 1. Sequence Listing <110> Dongpei Pharmaceutical Co., Ltd. <120> IL-17A binding peptide and its medical use <130> 19LG52E <140> PCT / EP2019 / 070265 <141> 2019-07-26 <150> EP18186029.7 <151> 2018-07-27 <160> 257 <170> BiSSAP 1.3.6 <210> 1 <211> 15 <212> PRT <213> Homo sapiens <400> 1 Ile His Val Thr Ile Pro Ala Asp Leu Trp Asp Trp Ile Asn Lys 1 5 10 15 <210> 2 <211> 15 <212> PRT <213> Homo sapiens <400> 2 Val His Val Thr Ile Pro Ala Asp Leu Trp Asp Trp Ile Asn Lys 1 5 10 15 <210> 3 <211> 15 <212> PRT <213> Homo sapiens <400> 3 Ile Met Val Thr Ile Pro Ala Asp Leu Trp Asp Trp Ile Asn Lys 1 5 10 15 <210> 4 <211> 15 <212> PRT <213> Homo sapiens <400> 4 Val Met Val Thr Ile Pro Ala Asp Leu Trp Asp Trp Ile Asn Lys 1 5 10 15 <210> 5 <211> 15 <212> PRT <213> Homo sapiens <400> 5 Ile His Val Thr Ile Pro Ala Glu Leu Trp Asp Trp Ile Asn Lys 1 5 10 15 <210> 6 <211> 15 <212> PRT <213> Homo sapiens <400> 6 Ile His Val Thr Ile Pro Ala Asp Leu Tyr Asp Trp Ile Asn Lys 1 5 10 15 <210> 7 <211> 15 <212> PRT <213> Homo sapiens <400> 7 Ile His Val Thr Ile Pro Ala Asp Leu Tyr Glu Trp Ile Asn Lys 1 5 10 15 <210> 8 <211> 15 <212> PRT <213> Homo sapiens <400> 8 Ile His Val Thr Ile Pro Ala Asp Leu Trp Asp Trp Val Asn Lys 1 5 10 15 <210> 9 <211> 15 <212> PRT <213> Homo sapiens <400> 9 Ile His Val Thr Ile Pro Ala Asp Leu Trp Asp Trp Ile Arg Lys 1 5 10 15 <210> 10 <211> 15 <212> PRT <213> Homo sapiens <400> 10 Ile His Val Thr Ile Pro Ala Asp Leu Trp Asp Trp Ile Asn Arg 1 5 10 15 <210> 11 <211> 15 <212> PRT <213> Homo sapiens <400> 11 Ile His Val Thr Ile Pro Ala Asp Leu Trp Asp Trp Ile Glu Lys 1 5 10 15 <210> 12 <211> 15 <212> PRT <213> Homo sapiens <400> 12 Ile His Val Thr Ile Pro Ala Asp Leu Trp Asp Trp Ile Asn Glu 1 5 10 15 <210> 13 <211> 15 <212> PRT <213> Homo sapiens <400> 13 Ile His Val Thr Ile Pro Ala Asp Leu Tyr Glu Trp Ile Asn Lys 1 5 10 15 <210> 14 <211> 15 <212> PRT <213> Homo sapiens <400> 14 Ile His Val Thr Ile Pro Ala Asp Leu Trp Asp Trp Val Arg Arg 1 5 10 15 <210> 15 <211> 15 <212> PRT <213> Homo sapiens <400> 15 Ile His Val Thr Ile Pro Ala Asp Leu Trp Asp Trp Val Glu Glu 1 5 10 15 <210> 16 <211> 15 <212> PRT <213> Homo sapiens <400> 16 Val Met Val Thr Ile Pro Ala Asp Leu Tyr Glu Trp Ile Asn Lys 1 5 10 15 <210> 17 <211> 15 <212> PRT <213> Homo sapiens <400> 17 Val Met Val Thr Ile Pro Ala Asp Leu Tyr Glu Trp Ile Arg Arg 1 5 10 15 <210> 18 <211> 15 <212> PRT <213> Homo sapiens <400> 18 Val Met Val Thr Ile Pro Ala Asp Leu Tyr Glu Trp Ile Glu Glu 1 5 10 15 <210> 19 <211> 21 <212> PRT <213> Homo sapiens <400> 19 Val Met Val Thr Ile Pro Ala Asp Leu Tyr Glu Trp Ile Glu Glu Arg 1 5 10 15 Arg Arg Arg Arg Arg 20 <210> 20 <211> 18 <212> PRT <213> Homo sapiens <400> 20 Val Met Val Thr Ile Pro Ala Asp Leu Tyr Glu Trp Ile Glu Glu Arg 1 5 10 15 Arg Arg <210> 21 <211> 23 <212> PRT <213> Homo sapiens <400> 21 Val Met Val Thr Ile Pro Ala Asp Leu Tyr Glu Trp Ile Glu Glu Ala 1 5 10 15 Arg Lys Lys Ala Ala Lys Ala 20 <210> 22 <211> 25 <212> PRT <213> Homo sapiens <400> 22 Val Met Val Thr Ile Pro Ala Asp Leu Tyr Glu Trp Ile Glu Glu Gly 1 5 10 15 Gly Thr Trp Trp Thr Glu Trp Ser Gln 20 25 <210> 23 <211> 22 <212> PRT <213> Homo sapiens <400> 23 Val Met Val Thr Ile Pro Ala Asp Leu Tyr Glu Trp Ile Glu Glu Thr 1 5 10 15 Trp Trp Glu Thr Trp Trp 20 <210> 24 <211> 20 <212> PRT <213> Homo sapiens <400> 24 Val Met Val Thr Ile Pro Ala Asp Leu Tyr Glu Trp Ile Glu Glu Val 1 5 10 15 Pro Gly Trp Gly 20 <210> 25 <211> 25 <212> PRT <213> Homo sapiens <400> 25 Val Met Val Thr Ile Pro Ala Asp Leu Tyr Glu Trp Ile Glu Glu Gly 1 5 10 15 Gly Lys Glu Thr Trp Trp Glu Thr Trp 20 25 <210> 26 <211> 20 <212> PRT <213> Homo sapiens <400> 26 Val Met Val Thr Ile Pro Ala Asp Leu Tyr Glu Trp Ile Glu Glu Val 1 5 10 15 Pro Gly Lys Gly 20 <210> 27 <211> 20 <212> PRT <213> Homo sapiens <400> 27 Val Met Val Thr Ile Pro Ala Asp Leu Tyr Glu Trp Ile Glu Glu Val 1 5 10 15 Pro Gly Ala Gly 20 <210> 28 <211> 31 <212> PRT <213> Homo sapiens <400> 28 Val Met Val Thr Ile Pro Ala Asp Leu Tyr Glu Trp Ile Glu Glu Arg 1 5 10 15 Gln Ile Lys Ile Trp Phe Gln Asn Arg Arg Met Lys Trp Lys Lys 20 25 30 <210> 29 <211> 23 <212> PRT <213> Homo sapiens <400> 29 Arg Arg Arg Arg Arg Arg Arg Arg Val Met Val Thr Ile Pro Ala Asp 1 5 10 15 Leu Tyr Glu Trp Ile Glu Glu 20 <210> 30 <211> 23 <212> PRT <213> Homo sapiens <400> 30 Val Met Val Thr Ile Pro Ala Asp Leu Tyr Glu Trp Ile Glu Glu Arg 1 5 10 15 Arg Arg Arg Arg Arg Arg Arg 20 <210> 31 <211> 23 <212> PRT <213> Homo sapiens <400> 31 Ala Arg Lys Lys Ala Ala Lys Ala Val Met Val Thr Ile Pro Ala Asp 1 5 10 15 Leu Tyr Glu Trp Ile Glu Glu 20 <210> 32 <211> 25 <212> PRT <213> Homo sapiens <400> 32 Gly Gly Thr Trp Trp Thr Glu Trp Ser Gln Val Met Val Thr Ile Pro 1 5 10 15 Ala Asp Leu Tyr Glu Trp Ile Glu Glu 20 25 <210> 33 <211> 25 <212> PRT <213> Homo sapiens <400> 33 Gly Gly Lys Glu Thr Trp Trp Glu Thr Trp Val Met Val Thr Ile Pro 1 5 10 15 Ala Asp Leu Tyr Glu Trp Ile Glu Glu 20 25 <210> 34 <211> 20 <212> PRT <213> Homo sapiens <400> 34 Val Pro Gly Trp Gly Val Met Val Thr Ile Pro Ala Asp Leu Tyr Glu 1 5 10 15 Trp Ile Glu Glu 20 <210> 35 <211> 20 <212> PRT <213> Homo sapiens <400> 35 Val Pro Gly Ala Gly Val Met Val Thr Ile Pro Ala Asp Leu Tyr Glu 1 5 10 15 Trp Ile Glu Glu 20 <210> 36 <211> 20 <212> PRT <213> Homo sapiens <400> 36 Val Pro Gly Lys Gly Val Met Val Thr Ile Pro Ala Asp Leu Tyr Glu 1 5 10 15 Trp Ile Glu Glu 20 <210> 37 <211> 25 <212> PRT <213> Homo sapiens <400> 37 Val Met Val Thr Ile Pro Ala Asp Leu Tyr Glu Trp Ile Glu Glu Val 1 5 10 15 Pro Gly Ala Gly Val Pro Gly Ala Gly 20 25 <210> 38 <211> 25 <212> PRT <213> Homo sapiens <400> 38 Val Pro Gly Ala Gly Val Pro Gly Ala Gly Val Met Val Thr Ile Pro 1 5 10 15 Ala Asp Leu Tyr Glu Trp Ile Glu Glu 20 25 <210> 39 <211> 20 <212> PRT <213> Homo sapiens <400> 39 Val Pro Gly Asp Gly Val Met Val Thr Ile Pro Ala Asp Leu Tyr Glu 1 5 10 15 Trp Ile Glu Glu 20 <210> 40 <211> 20 <212> PRT <213> Homo sapiens <400> 40 Val Met Val Thr Ile Pro Ala Asp Leu Tyr Glu Trp Ile Glu Glu Val 1 5 10 15 Pro Gly Asp Gly 20 <210> 41 <211> 16 <212> PRT <213> Homo sapiens <400> 41 Asp Leu Ser Ala Val Cys Trp Ala Phe Pro Trp Asp Pro Glu Cys His 1 5 10 15 <210> 42 <211> 15 <212> PRT <213> Homo sapiens <400> 42 Ile Met Val Thr Ile Pro Ala Asp Leu Tyr Glu Trp Ile Glu Glu 1 5 10 15 <210> 43 <211> 16 <212> PRT <213> Homo sapiens <400> 43 Ile Met Val Thr Ile Pro Ala Asp Leu Tyr Glu Trp Ile Glu Glu Gln 1 5 10 15 <210> 44 <211> 8 <212> PRT <213> Homo sapiens <400> 44 Ala Arg Lys Lys Ala Ala Lys Ala 1 5 <210> 45 <211> 8 <212> PRT <213> Homo sapiens <400> 45 Thr Trp Trp Thr Glu Trp Ser Gln 1 5 <210> 46 <211> 7 <212> PRT <213> Homo sapiens <400> 46 Thr Trp Trp Glu Thr Trp Trp 1 5 <210> 47 <211> 5 <212> PRT <213> Homo sapiens <400> 47 Val Pro Gly Trp Gly 1 5 <210> 48 <211> 8 <212> PRT <213> Homo sapiens <400> 48 Lys Glu Thr Trp Trp Glu Thr Trp 1 5 <210> 49 <211> 5 <212> PRT <213> Homo sapiens <400> 49 Val Pro Gly Lys Gly 1 5 <210> 50 <211> 5 <212> PRT <213> Homo sapiens <400> 50 Val Pro Gly Ala Gly 1 5 <210> 51 <211> 16 <212> PRT <213> Homo sapiens <400> 51 Arg Gln Ile Lys Ile Trp Phe Gln Asn Arg Arg Met Lys Trp Lys Lys 1 5 10 15 <210> 52 <211> 8 <212> PRT <213> Homo sapiens <400> 52 Arg Arg Arg Arg Arg Arg Arg Arg 1 5 <210> 53 <211> 5 <212> PRT <213> Homo sapiens <400> 53 Val Pro Gly Asp Gly 1 5 <210> 54 <211> 5 <212> PRT <213> Artificial sequence <220> <223> Synthetically produced peptides <220> <221> Variants <222> 4 <223> Xaa = any amino acid <400> 54 Val Pro Gly Xaa Gly 1 5 <210> 55 <211> 4 <212> PRT <213> Homo sapiens <400> 55 Ile Pro Gly Gly 1 <210> 56 <211> 5 <212> PRT <213> Homo sapiens <400> 56 Ala Val Gly Val Pro 1 5 <210> 57 <211> 5 <212> PRT <213> Artificial sequence <220> <223> Synthetically produced peptides <220> <221> Variants <222> 4 <223> Xaa = any amino acid <400> 57 Ile Pro Gly Xaa Gly 1 5 <210> 58 <211> 5 <212> PRT <213> Homo sapiens <400> 58 Ile Pro Gly Val Gly 1 5 <210> 59 <211> 5 <212> PRT <213> Artificial sequence <220> <223> Synthetically produced peptides <220> <221> Variants <222> 4 <223> Xaa = any amino acid <400> 59 Leu Pro Gly Xaa Gly 1 5 <210> 60 <211> 5 <212> PRT <213> Homo sapiens <400> 60 Leu Pro Gly Val Gly 1 5 <210> 61 <211> 6 <212> PRT <213> Homo sapiens <400> 61 Val Ala Pro Gly Val Gly 1 5 <210> 62 <211> 5 <212> PRT <213> Artificial sequence <220> <223> Synthetically produced peptides <220> <221> Variants <222> 1 <223> Xaa = any amino acid <400> 62 Xaa Pro Gly Val Gly 1 5 <210> 63 <211> 8 <212> PRT <213> Homo sapiens <400> 63 Gly Val Gly Val Pro Gly Val Gly 1 5 <210> 64 <211> 9 <212> PRT <213> Homo sapiens <400> 64 Val Pro Gly Phe Gly Val Gly Ala Gly 1 5 <210> 65 <211> 8 <212> PRT <213> Homo sapiens <400> 65 Val Pro Gly Gly Val Pro Gly Gly 1 5 <210> 66 <211> 15 <212> PRT <213> Homo sapiens <400> 66 Ile Val Val Thr Met Pro Ala Asp Leu Trp Asp Trp Ile Lys Ala 1 5 10 15 <210> 67 <211> 15 <212> PRT <213> Homo sapiens <400> 67 Ile Val Val Thr Met Pro Ala Asp Leu Trp Asp Trp Ile Arg Ala 1 5 10 15 <210> 68 <211> 15 <212> PRT <213> Homo sapiens <400> 68 Ile Val Val Thr Met Pro Ala Asp Leu Trp Asp Trp Ile Arg Lys 1 5 10 15 <210> 69 <211> 15 <212> PRT <213> Homo sapiens <400> 69 Ile Val Val Thr Met Pro Ala Asp Leu Trp Asp Trp Ile Ala Ala 1 5 10 15 <210> 70 <211> 15 <212> PRT <213> Homo sapiens <400> 70 Ile Val Val Thr Met Pro Ala Asp Leu Trp Asp Trp Ala Arg Ala 1 5 10 15 <210> 71 <211> 15 <212> PRT <213> Homo sapiens <400> 71 Ile Val Val Thr Met Pro Ala Asp Leu Trp Ala Trp Ile Arg Ala 1 5 10 15 <210> 72 <211> 15 <212> PRT <213> Homo sapiens <400> 72 Ile Val Val Thr Met Pro Ala Asp Leu Ala Asp Trp Ile Arg Ala 1 5 10 15 <210> 73 <211> 15 <212> PRT <213> Homo sapiens <400> 73 Ile Val Val Thr Met Pro Ala Asp Ala Trp Asp Trp Ile Arg Ala 1 5 10 15 <210> 74 <211> 15 <212> PRT <213> Homo sapiens <400> 74 Ile Val Val Thr Met Pro Ala Asp Leu Trp Asp Trp Ile Arg Ala 1 5 10 15 <210> 75 <211> 15 <212> PRT <213> Homo sapiens <400> 75 Ile Val Val Thr Ala Pro Ala Asp Leu Trp Asp Trp Ile Arg Ala 1 5 10 15 <210> 76 <211> 15 <212> PRT <213> Homo sapiens <400> 76 Ile Val Val Ala Met Pro Ala Asp Leu Trp Asp Trp Ile Arg Ala 1 5 10 15 <210> 77 <211> 15 <212> PRT <213> Homo sapiens <400> 77 Ile Ala Val Thr Met Pro Ala Asp Leu Trp Asp Trp Ile Arg Ala 1 5 10 15 <210> 78 <211> 15 <212> PRT <213> Homo sapiens <400> 78 Ala Val Val Thr Met Pro Ala Asp Leu Trp Asp Trp Ile Arg Ala 1 5 10 15 <210> 79 <211> 15 <212> PRT <213> Homo sapiens <400> 79 Ile His Val Thr Met Pro Ala Asp Leu Trp Asp Trp Ile Arg Ala 1 5 10 15 <210> 80 <211> 15 <212> PRT <213> Homo sapiens <400> 80 Ile Gln Val Thr Met Pro Ala Asp Leu Trp Asp Trp Ile Arg Ala 1 5 10 15 <210> 81 <211> 15 <212> PRT <213> Homo sapiens <400> 81 Ile Arg Val Thr Met Pro Ala Asp Leu Trp Asp Trp Ile Arg Ala 1 5 10 15 <210> 82 <211> 15 <212> PRT <213> Homo sapiens <400> 82 Ile Thr Val Thr Met Pro Ala Asp Leu Trp Asp Trp Ile Arg Ala 1 5 10 15 <210> 83 <211> 15 <212> PRT <213> Homo sapiens <400> 83 Ile Trp Val Thr Met Pro Ala Asp Leu Trp Asp Trp Ile Arg Ala 1 5 10 15 <210> 84 <211> 15 <212> PRT <213> Homo sapiens <400> 84 Ile Tyr Val Thr Met Pro Ala Asp Leu Trp Asp Trp Ile Arg Ala 1 5 10 15 <210> 85 <211> 15 <212> PRT <213> Homo sapiens <400> 85 Ile Val Val Thr Ile Pro Ala Asp Leu Trp Asp Trp Ile Arg Ala 1 5 10 15 <210> 86 <211> 15 <212> PRT <213> Homo sapiens <400> 86 Ile Val Val Thr Leu Pro Ala Asp Leu Trp Asp Trp Ile Arg Ala 1 5 10 15 <210> 87 <211> 15 <212> PRT <213> Homo sapiens <400> 87 Ile Val Val Thr Val Pro Ala Asp Leu Trp Asp Trp Ile Arg Ala 1 5 10 15 <210> 88 <211> 15 <212> PRT <213> Homo sapiens <400> 88 Ile Val Val Thr Met Pro Ala Asp Leu Trp Asp Trp Ile Met Ala 1 5 10 15 <210> 89 <211> 15 <212> PRT <213> Homo sapiens <400> 89 Ile Val Val Thr Met Pro Ala Asp Leu Trp Asp Trp Ile Asn Ala 1 5 10 15 <210> 90 <211> 15 <212> PRT <213> Homo sapiens <400> 90 Ile Val Val Thr Met Pro Ala Asp Leu Trp Asp Trp Ile Gln Ala 1 5 10 15 <210> 91 <211> 15 <212> PRT <213> Homo sapiens <400> 91 Ile Val Val Thr Ile Pro Ala Asp Leu Trp Asp Trp Ile Arg Ala 1 5 10 15 <210> 92 <211> 15 <212> PRT <213> Homo sapiens <400> 92 Ile Val Val Thr Leu Pro Ala Asp Leu Trp Asp Trp Ile Arg Ala 1 5 10 15 <210> 93 <211> 15 <212> PRT <213> Homo sapiens <400> 93 Ile His Val Thr Ile Pro Ala Asp Leu Trp Asp Trp Ile Asn Lys 1 5 10 15 <210> 94 <211> 14 <212> PRT <213> Homo sapiens <400> 94 Ile His Val Thr Ile Pro Ala Asp Leu Trp Asp Trp Ile Asn 1 5 10 <210> 95 <211> 13 <212> PRT <213> Homo sapiens <400> 95 Ile His Val Thr Ile Pro Ala Asp Leu Trp Asp Trp Ile 1 5 10 <210> 96 <211> 20 <212> PRT <213> Homo sapiens <400> 96 Asp Ser Ser Ala Val Cys Trp Ala Phe Pro His His Pro Leu Cys His 1 5 10 15 Met Lys Ala Thr 20 <210> 97 <211> 16 <212> PRT <213> Homo sapiens <400> 97 Ala Asp Ala Asp Met Cys Trp Phe Phe Pro Thr Ser Pro Trp Cys His 1 5 10 15 <210> 98 <211> 17 <212> PRT <213> Homo sapiens <400> 98 Asp Leu Ser Ala Val Cys Trp Ala Phe Pro Trp Asp Pro Glu Cys His 1 5 10 15 Met <210> 99 <211> 16 <212> PRT <213> Homo sapiens <400> 99 Asp Ser Ser Ala Val Cys Trp Ala Phe Pro Tyr Leu Pro Glu Cys His 1 5 10 15 <210> 100 <211> 16 <212> PRT <213> Homo sapiens <400> 100 Asp Ile Ser Ala Val Cys Trp Ala Phe Pro Phe Asp Pro Glu Cys His 1 5 10 15 <210> 101 <211> 20 <212> PRT <213> Homo sapiens <400> 101 Ala Tyr Glu Cys Pro Arg Leu Glu Tyr Asp Met Phe Gly Ala Leu His 1 5 10 15 Cys Leu Pro Ser 20 <210> 102 <211> 15 <212> PRT <213> Homo sapiens <400> 102 Cys Pro Arg Leu Glu Tyr Asp Met Phe Gly Ala Leu His Cys Leu 1 5 10 15 <210> 103 <211> 15 <212> PRT <213> Homo sapiens <400> 103 Cys Leu Asp Leu Gln Tyr Asp Pro Trp Gly Ala Leu His Cys Ile 1 5 10 15 <210> 104 <211> 15 <212> PRT <213> Homo sapiens <400> 104 Cys Phe Asp Leu Gln Tyr Asp Pro Trp Gly Ala Leu His Cys Ile 1 5 10 15 <210> 105 <211> 15 <212> PRT <213> Homo sapiens <400> 105 Cys Leu Asp Leu Gln Tyr Asp Met Phe Gly Ala Leu His Cys Val 1 5 10 15 <210> 106 <211> 15 <212> PRT <213> Homo sapiens <400> 106 Cys Leu Asp Leu Val Tyr Asp Pro Trp Gly Ala Leu His Cys Ile 1 5 10 15 <210> 107 <211> 15 <212> PRT <213> Homo sapiens <400> 107 Cys Trp Val Leu Glu Tyr Asp Met Phe Gly Ala Leu His Cys Arg 1 5 10 15 <210> 108 <211> 15 <212> PRT <213> Homo sapiens <400> 108 Cys Trp Ala Leu Glu Tyr Asp Met Phe Gly Tyr Leu His Cys Arg 1 5 10 15 <210> 109 <211> 15 <212> PRT <213> Homo sapiens <400> 109 Cys Trp Val Leu Glu Tyr Asp Met Phe Gly Phe Leu His Cys Arg 1 5 10 15 <210> 110 <211> 15 <212> PRT <213> Homo sapiens <400> 110 Cys Trp Val Leu Glu Tyr Asp Met Phe Gly Tyr Leu His Cys Arg 1 5 10 15 <210> 111 <211> 15 <212> PRT <213> Homo sapiens <400> 111 Val His Val Thr Ile Pro Ala Glu Leu Trp Glu Trp Val Arg Arg 1 5 10 15 <210> 112 <211> 15 <212> PRT <213> Homo sapiens <400> 112 Val His Phe Thr Ile Pro Ala Asp Leu Trp Glu Trp Val Arg Arg 1 5 10 15 <210> 113 <211> 15 <212> PRT <213> Homo sapiens <400> 113 Val His Val Gln Ile Pro Ala Asp Leu Trp Glu Trp Val Arg Arg 1 5 10 15 <210> 114 <211> 15 <212> PRT <213> Homo sapiens <400> 114 Val His Val Thr Phe Pro Ala Asp Leu Trp Glu Trp Val Arg Arg 1 5 10 15 <210> 115 <211> 15 <212> PRT <213> Homo sapiens <400> 115 Val His Val Thr Ile Pro Gln Asp Leu Trp Glu Trp Val Arg Arg 1 5 10 15 <210> 116 <211> 15 <212> PRT <213> Homo sapiens <400> 116 Val His Val Thr Ile Pro Ala Asn Leu Trp Glu Trp Val Arg Arg 1 5 10 15 <210> 117 <211> 15 <212> PRT <213> Homo sapiens <400> 117 Val His Val Thr Ile Pro Ala Asp Phe Trp Glu Trp Val Arg Arg 1 5 10 15 <210> 118 <211> 15 <212> PRT <213> Homo sapiens <400> 118 Val His Val Thr Ile Pro Ala Asp Leu Tyr Glu Trp Val Arg Arg 1 5 10 15 <210> 119 <211> 15 <212> PRT <213> Homo sapiens <400> 119 Val His Val Thr Ile Pro Ala Asp Leu Trp Asn Trp Val Arg Arg 1 5 10 15 <210> 120 <211> 15 <212> PRT <213> Homo sapiens <400> 120 Val His Val Thr Ile Pro Ala Asp Leu Trp Glu Phe Val Arg Arg 1 5 10 15 <210> 121 <211> 15 <212> PRT <213> Homo sapiens <400> 121 Val His Val Thr Ile Pro Ala Asp Leu Trp Glu Trp Phe Arg Arg 1 5 10 15 <210> 122 <211> 15 <212> PRT <213> Homo sapiens <400> 122 Val His Val Tyr Ile Pro Ala Glu Leu Trp Glu Trp Val Arg Arg 1 5 10 15 <210> 123 <211> 15 <212> PRT <213> Homo sapiens <400> 123 Val His Val Thr Ile Pro Ala Glu Trp Trp Glu Trp Val Arg Arg 1 5 10 15 <210> 124 <211> 15 <212> PRT <213> Homo sapiens <400> 124 Val His Phe Thr Phe Pro Gln Asp Leu Trp Glu Trp Val Arg Arg 1 5 10 15 <210> 125 <211> 15 <212> PRT <213> Homo sapiens <400> 125 Val His Phe Thr Phe Pro Gln Asp Phe Trp Glu Trp Val Arg Arg 1 5 10 15 <210> 126 <211> 15 <212> PRT <213> Homo sapiens <400> 126 Val His Phe Thr Ile Pro Gln Asp Leu Tyr Glu Trp Val Arg Arg 1 5 10 15 <210> 127 <211> 15 <212> PRT <213> Homo sapiens <400> 127 Val His Phe Thr Phe Pro Gln Asp Leu Trp Asn Trp Val Arg Arg 1 5 10 15 <210> 128 <211> 15 <212> PRT <213> Homo sapiens <400> 128 Val His Phe Thr Phe Pro Gln Asp Leu Trp Glu Phe Val Arg Arg 1 5 10 15 <210> 129 <211> 15 <212> PRT <213> Homo sapiens <400> 129 Val His Phe Thr Phe Pro Gln Asp Leu Trp Glu Trp Phe Arg Arg 1 5 10 15 <210> 130 <211> 15 <212> PRT <213> Homo sapiens <400> 130 Val His Phe Gln Phe Pro Ala Asp Leu Trp Glu Trp Val Arg Arg 1 5 10 15 <210> 131 <211> 15 <212> PRT <213> Homo sapiens <400> 131 Val His Phe Gln Phe Pro Ala Asp Phe Trp Glu Trp Val Arg Arg 1 5 10 15 <210> 132 <211> 15 <212> PRT <213> Homo sapiens <400> 132 Val His Phe Gln Phe Pro Ala Asp Leu Tyr Glu Trp Val Arg Arg 1 5 10 15 <210> 133 <211> 15 <212> PRT <213> Homo sapiens <400> 133 Val His Phe Gln Phe Pro Ala Asp Leu Trp Asn Trp Val Arg Arg 1 5 10 15 <210> 134 <211> 15 <212> PRT <213> Homo sapiens <400> 134 Val His Phe Gln Phe Pro Ala Asp Leu Trp Glu Phe Val Arg Arg 1 5 10 15 <210> 135 <211> 15 <212> PRT <213> Homo sapiens <400> 135 Val His Phe Gln Phe Pro Ala Asp Leu Trp Glu Trp Phe Arg Arg 1 5 10 15 <210> 136 <211> 15 <212> PRT <213> Homo sapiens <400> 136 Val His Phe Gln Phe Pro Gln Asp Trp Trp Glu Trp Val Arg Arg 1 5 10 15 <210> 137 <211> 15 <212> PRT <213> Homo sapiens <400> 137 Val His Phe Gln Ile Pro Gln Asp Trp Trp Glu Trp Val Arg Arg 1 5 10 15 <210> 138 <211> 15 <212> PRT <213> Homo sapiens <400> 138 Val His Phe Gln Ile Phe Gln Asp Trp Trp Glu Trp Val Arg Arg 1 5 10 15 <210> 139 <211> 15 <212> PRT <213> Homo sapiens <400> 139 Val His Phe Gln Phe Pro Gln Asp Trp Trp Asn Trp Val Arg Arg 1 5 10 15 <210> 140 <211> 15 <212> PRT <213> Homo sapiens <400> 140 Val His Phe Gln Phe Pro Gln Asp Leu Trp Glu Phe Val Arg Arg 1 5 10 15 <210> 141 <211> 15 <212> PRT <213> Homo sapiens <400> 141 Val His Phe Gln Phe Pro Gln Asp Trp Trp Glu Trp Phe Arg Arg 1 5 10 15 <210> 142 <211> 15 <212> PRT <213> Homo sapiens <400> 142 Val His Phe Thr Ile Pro Ala Asp Phe Trp Glu Trp Phe Arg Arg 1 5 10 15 <210> 143 <211> 15 <212> PRT <213> Homo sapiens <400> 143 Val His Val Gln Ile Pro Ala Asp Phe Trp Glu Trp Phe Arg Arg 1 5 10 15 <210> 144 <211> 15 <212> PRT <213> Homo sapiens <400> 144 Val His Val Thr Phe Pro Ala Asp Leu Trp Glu Trp Phe Arg Arg 1 5 10 15 <210> 145 <211> 15 <212> PRT <213> Homo sapiens <400> 145 Val His Val Thr Ile Pro Gln Asp Phe Trp Glu Trp Phe Arg Arg 1 5 10 15 <210> 146 <211> 15 <212> PRT <213> Homo sapiens <400> 146 Val His Phe Thr Ile Pro Gln Asp Trp Trp Glu Trp Val Arg Arg 1 5 10 15 <210> 147 <211> 15 <212> PRT <213> Homo sapiens <400> 147 Val His Phe Thr Phe Pro Gln Asp Leu Tyr Asn Trp Val Arg Arg 1 5 10 15 <210> 148 <211> 15 <212> PRT <213> Homo sapiens <400> 148 Val His Phe Thr Phe Pro Gln Asp Leu Tyr Asn Phe Val Arg Arg 1 5 10 15 <210> 149 <211> 15 <212> PRT <213> intelligent person <400> 149 Val His Val Thr Ile Pro Ala Asp Leu Tyr Asn Phe Phe Arg Arg 1 5 10 15 <210> 150 <211> 15 <212> PRT <213> intelligent person <400> 150 Val His Phe Gln Phe Pro Gln Asp Leu Trp Glu Trp Val Arg Arg 1 5 10 15 <210> 151 <211> 14 <212> PRT <213> intelligent person <400> 151 Val His Phe Thr Ile Pro Gln Asp Leu Tyr Asn Trp Arg Arg 1 5 10 <210> 152 <211> 15 <212> PRT <213> intelligent person <400> 152 Val His Phe Thr Ile Pro Ala Asp Leu Tyr Asn Phe Val Arg Arg 1 5 10 15 <210> 153 <211> 15 <212> PRT <213> intelligent person <400> 153 Val His Phe Gln Ile Pro Gln Asp Leu Tyr Asn Phe Phe Arg Arg 1 5 10 15 <210> 154 <211> 15 <212> PRT <213> Homo sapiens <400> 154 Val His Phe Gln Phe Pro Gln Glu Trp Tyr Asn Trp Phe Arg Arg 1 5 10 15 <210> 155 <211> 15 <212> PRT <213> Homo sapiens <400> 155 Val Arg Phe Gln Phe Gly Gln Glu Trp Tyr Asn Phe Phe Arg Arg 1 5 10 15 <210> 156 <211> 15 <212> PRT <213> Homo sapiens <400> 156 Val Pro Gly Ala Gly Val Pro Gly Ala Gly Ile His Val Thr Ile 1 5 10 15 <210> 157 <211> 17 <212> PRT <213> Homo sapiens <400> 157 Val Pro Gly Ala Gly Val Pro Gly Ala Gly Ile His Val Thr Ile Pro 1 5 10 15 Ala <210> 158 <211> 15 <212> PRT <213> Homo sapiens <400> 158 Ala His Val Thr Ile Pro Ala Asp Leu Trp Asp Trp Ile Asn Lys 1 5 10 15 <210> 159 <211> 15 <212> PRT <213> Homo sapiens <400> 159 Gly His Val Thr Ile Pro Ala Asp Leu Trp Asp Trp Ile Asn Lys 1 5 10 15 <210> 160 <211> 15 <212> PRT <213> Homo sapiens <400> 160 Leu His Val Thr Ile Pro Ala Asp Leu Trp Asp Trp Ile Asn Lys 1 5 10 15 <210> 161 <211> 15 <212> PRT <213> Homo sapiens <400> 161 Pro His Val Thr Ile Pro Ala Asp Leu Trp Asp Trp Ile Asn Lys 1 5 10 15 <210> 162 <211> 15 <212> PRT <213> Homo sapiens <400> 162 Ile Arg Val Thr Ile Pro Ala Asp Leu Trp Asp Trp Ile Asn Lys 1 5 10 15 <210> 163 <211> 15 <212> PRT <213> Homo sapiens <400> 163 Ile Lys Val Thr Ile Pro Ala Asp Leu Trp Asp Trp Ile Asn Lys 1 5 10 15 <210> 164 <211> 15 <212> PRT <213> Homo sapiens <400> 164 Ile Glu Val Thr Ile Pro Ala Asp Leu Trp Asp Trp Ile Asn Lys 1 5 10 15 <210> 165 <211> 15 <212> PRT <213> Homo sapiens <400> 165 Ile Gln Val Thr Ile Pro Ala Asp Leu Trp Asp Trp Ile Asn Lys 1 5 10 15 <210> 166 <211> 15 <212> PRT <213> Homo sapiens <400> 166 Ile Tyr Val Thr Ile Pro Ala Asp Leu Trp Asp Trp Ile Asn Lys 1 5 10 15 <210> 167 <211> 15 <212> PRT <213> Homo sapiens <400> 167 Ile His Ala Thr Ile Pro Ala Asp Leu Trp Asp Trp Ile Asn Lys 1 5 10 15 <210> 168 <211> 15 <212> PRT <213> Homo sapiens <400> 168 Ile His Gly Thr Ile Pro Ala Asp Leu Trp Asp Trp Ile Asn Lys 1 5 10 15 <210> 169 <211> 15 <212> PRT <213> Homo sapiens <400> 169 Ile His Leu Thr Ile Pro Ala Asp Leu Trp Asp Trp Ile Asn Lys 1 5 10 15 <210> 170 <211> 15 <212> PRT <213> Homo sapiens <400> 170 Ile His Pro Thr Ile Pro Ala Asp Leu Trp Asp Trp Ile Asn Lys 1 5 10 15 <210> 171 <211> 15 <212> PRT <213> Homo sapiens <400> 171 Ile His Ile Thr Ile Pro Ala Asp Leu Trp Asp Trp Ile Asn Lys 1 5 10 15 <210> 172 <211> 15 <212> PRT <213> Homo sapiens <400> 172 Ile His Tyr Thr Ile Pro Ala Asp Leu Trp Asp Trp Ile Asn Lys 1 5 10 15 <210> 173 <211> 15 <212> PRT <213> Homo sapiens <400> 173 Ile His Trp Thr Ile Pro Ala Asp Leu Trp Asp Trp Ile Asn Lys 1 5 10 15 <210> 174 <211> 15 <212> PRT <213> Homo sapiens <400> 174 Ile His Phe Thr Ile Pro Ala Asp Leu Trp Asp Trp Ile Asn Lys 1 5 10 15 <210> 175 <211> 15 <212> PRT <213> Homo sapiens <400> 175 Ile His Val Ser Ile Pro Ala Asp Leu Trp Asp Trp Ile Asn Lys 1 5 10 15 <210> 176 <211> 15 <212> PRT <213> Homo sapiens <400> 176 Ile His Val Tyr Ile Pro Ala Asp Leu Trp Asp Trp Ile Asn Lys 1 5 10 15 <210> 177 <211> 15 <212> PRT <213> Homo sapiens <400> 177 Ile His Val Asn Ile Pro Ala Asp Leu Trp Asp Trp Ile Asn Lys 1 5 10 15 <210> 178 <211> 15 <212> PRT <213> Homo sapiens <400> 178 Ile His Val Gln Ile Pro Ala Asp Leu Trp Asp Trp Ile Asn Lys 1 5 10 15 <210> 179 <211> 15 <212> PRT <213> Homo sapiens <400> 179 Ile His Val Thr Ala Pro Ala Asp Leu Trp Asp Trp Ile Asn Lys 1 5 10 15 <210> 180 <211> 15 <212> PRT <213> Homo sapiens <400> 180 Ile His Val Thr Gly Pro Ala Asp Leu Trp Asp Trp Ile Asn Lys 1 5 10 15 <210> 181 <211> 15 <212> PRT <213> Homo sapiens <400> 181 Ile His Val Thr Leu Pro Ala Asp Leu Trp Asp Trp Ile Asn Lys 1 5 10 15 <210> 182 <211> 15 <212> PRT <213> Homo sapiens <400> 182 Ile His Val Thr Pro Pro Ala Asp Leu Trp Asp Trp Ile Asn Lys 1 5 10 15 <210> 183 <211> 15 <212> PRT <213> Homo sapiens <400> 183 Ile His Val Thr Val Pro Ala Asp Leu Trp Asp Trp Ile Asn Lys 1 5 10 15 <210> 184 <211> 15 <212> PRT <213> Homo sapiens <400> 184 Ile His Val Thr Phe Pro Ala Asp Leu Trp Asp Trp Ile Asn Lys 1 5 10 15 <210> 185 <211> 15 <212> PRT <213> Homo sapiens <400> 185 Ile His Val Thr Tyr Pro Ala Asp Leu Trp Asp Trp Ile Asn Lys 1 5 10 15 <210> 186 <211> 15 <212> PRT <213> Homo sapiens <400> 186 Ile His Val Thr Ile Ala Ala Asp Leu Trp Asp Trp Ile Asn Lys 1 5 10 15 <210> 187 <211> 15 <212> PRT <213> Homo sapiens <400> 187 Ile His Val Thr Ile Gly Ala Asp Leu Trp Asp Trp Ile Asn Lys 1 5 10 15 <210> 188 <211> 15 <212> PRT <213> Homo sapiens <400> 188 Ile His Val Thr Ile Leu Ala Asp Leu Trp Asp Trp Ile Asn Lys 1 5 10 15 <210> 189 <211> 15 <212> PRT <213> Homo sapiens <400> 189 Ile His Val Thr Ile Val Ala Asp Leu Trp Asp Trp Ile Asn Lys 1 5 10 15 <210> 190 <211> 15 <212> PRT <213> Homo sapiens <400> 190 Ile His Val Thr Ile Ile Ala Asp Leu Trp Asp Trp Ile Asn Lys 1 5 10 15 <210> 191 <211> 15 <212> PRT <213> Homo sapiens <400> 191 Ile His Val Thr Ile Asn Ala Asp Leu Trp Asp Trp Ile Asn Lys 1 5 10 15 <210> 192 <211> 15 <212> PRT <213> Homo sapiens <400> 192 Ile His Val Thr Ile Pro Gly Asp Leu Trp Asp Trp Ile Asn Lys 1 5 10 15 <210> 193 <211> 15 <212> PRT <213> Homo sapiens <400> 193 Ile His Val Thr Ile Pro Leu Asp Leu Trp Asp Trp Ile Asn Lys 1 5 10 15 <210> 194 <211> 15 <212> PRT <213> Homo sapiens <400> 194 Ile His Val Thr Ile Pro Pro Asp Leu Trp Asp Trp Ile Asn Lys 1 5 10 15 <210> 195 <211> 15 <212> PRT <213> Homo sapiens <400> 195 Ile His Val Thr Ile Pro Val Asp Leu Trp Asp Trp Ile Asn Lys 1 5 10 15 <210> 196 <211> 15 <212> PRT <213> Homo sapiens <400> 196 Ile His Val Thr Ile Pro Ile Asp Leu Trp Asp Trp Ile Asn Lys 1 5 10 15 <210> 197 <211> 15 <212> PRT <213> Homo sapiens <400> 197 Ile His Val Thr Ile Pro Asn Asp Leu Trp Asp Trp Ile Asn Lys 1 5 10 15 <210> 198 <211> 15 <212> PRT <213> Homo sapiens <400> 198 Ile His Val Thr Ile Pro Gln Asp Leu Trp Asp Trp Ile Asn Lys 1 5 10 15 <210> 199 <211> 15 <212> PRT <213> Homo sapiens <400> 199 Ile His Val Thr Ile Pro Ala Gln Leu Trp Asp Trp Ile Asn Lys 1 5 10 15 <210> 200 <211> 15 <212> PRT <213> Homo sapiens <400> 200 Ile His Val Thr Ile Pro Ala Tyr Leu Trp Asp Trp Ile Asn Lys 1 5 10 15 <210> 201 <211> 15 <212> PRT <213> Homo sapiens <400> 201 Ile His Val Thr Ile Pro Ala Asp Ala Trp Asp Trp Ile Asn Lys 1 5 10 15 <210> 202 <211> 15 <212> PRT <213> Homo sapiens <400> 202 Ile His Val Thr Ile Pro Ala Asp Gly Trp Asp Trp Ile Asn Lys 1 5 10 15 <210> 203 <211> 15 <212> PRT <213> Homo sapiens <400> 203 Ile His Val Thr Ile Pro Ala Asp Pro Trp Asp Trp Ile Asn Lys 1 5 10 15 <210> 204 <211> 15 <212> PRT <213> Homo sapiens <400> 204 Ile His Val Thr Ile Pro Ala Asp Val Trp Asp Trp Ile Asn Lys 1 5 10 15 <210> 205 <211> 15 <212> PRT <213> Homo sapiens <400> 205 Ile His Val Thr Ile Pro Ala Asp Ile Trp Asp Trp Ile Asn Lys 1 5 10 15 <210> 206 <211> 15 <212> PRT <213> Homo sapiens <400> 206 Ile His Val Thr Ile Pro Ala Asp Leu His Asp Trp Ile Asn Lys 1 5 10 15 <210> 207 <211> 15 <212> PRT <213> Homo sapiens <400> 207 Ile His Val Thr Ile Pro Ala Asp Leu Phe Asp Trp Ile Asn Lys 1 5 10 15 <210> 208 <211> 15 <212> PRT <213> Homo sapiens <400> 208 Ile His Val Thr Ile Pro Ala Asp Leu Trp Asn Trp Ile Asn Lys 1 5 10 15 <210> 209 <211> 15 <212> PRT <213> Homo sapiens <400> 209 Ile His Val Thr Ile Pro Ala Asp Leu Trp Asp His Ile Asn Lys 1 5 10 15 <210> 210 <211> 15 <212> PRT <213> Homo sapiens <400> 210 Ile His Val Thr Ile Pro Ala Asp Leu Trp Asp Phe Ile Asn Lys 1 5 10 15 <210> 211 <211> 15 <212> PRT <213> Homo sapiens <400> 211 Ile His Val Thr Ile Pro Ala Asp Leu Trp Asp Tyr Ile Asn Lys 1 5 10 15 <210> 212 <211> 15 <212> PRT <213> Homo sapiens <400> 212 Ile His Val Thr Ile Pro Ala Asp Leu Trp Asp Trp Glu Asn Lys 1 5 10 15 <210> 213 <211> 15 <212> PRT <213> Homo sapiens <400> 213 Ile His Val Thr Ile Pro Ala Asp Leu Trp Asp Trp Lys Asn Lys 1 5 10 15 <210> 214 <211> 15 <212> PRT <213> Homo sapiens <400> 214 Ile His Val Thr Ile Pro Ala Asp Leu Trp Asp Trp Ala Asn Lys 1 5 10 15 <210> 215 <211> 15 <212> PRT <213> Homo sapiens <400> 215 Ile His Val Thr Ile Pro Ala Asp Leu Trp Asp Trp Gly Asn Lys 1 5 10 15 <210> 216 <211> 15 <212> PRT <213> Homo sapiens <400> 216 Ile His Val Thr Ile Pro Ala Asp Leu Trp Asp Trp Leu Asn Lys 1 5 10 15 <210> 217 <211> 15 <212> PRT <213> Homo sapiens <400> 217 Ile His Val Thr Ile Pro Ala Asp Leu Trp Asp Trp Pro Asn Lys 1 5 10 15 <210> 218 <211> 15 <212> PRT <213> Homo sapiens <400> 218 Ile His Val Thr Ile Pro Ala Asp Leu Trp Asp Trp Phe Asn Lys 1 5 10 15 <210> 219 <211> 15 <212> PRT <213> Homo sapiens <400> 219 Ile His Val Thr Ile Pro Ala Asp Leu Trp Asp Trp Tyr Asn Lys 1 5 10 15 <210> 220 <211> 15 <212> PRT <213> Homo sapiens <400> 220 Ile His Val Thr Ile Pro Ala Asp Leu Trp Asp Trp Ile Gln Lys 1 5 10 15 <210> 221 <211> 15 <212> PRT <213> Homo sapiens <400> 221 Ile His Val Thr Ile Pro Ala Asp Leu Trp Asp Trp Ile Asp Lys 1 5 10 15 <210> 222 <211> 15 <212> PRT <213> Homo sapiens <400> 222 Ile His Val Thr Ile Pro Ala Asp Leu Trp Asp Trp Ile Asn His 1 5 10 15 <210> 223 <211> 15 <212> PRT <213> Homo sapiens <400> 223 Ile His Val Thr Ile Pro Ala Asp Leu Trp Asp Trp Ile Asn Asp 1 5 10 15 <210> 224 <211> 15 <212> PRT <213> Homo sapiens <400> 224 Val His Val Thr Val Pro Gln Glu Leu Trp Glu Trp Val Arg Arg 1 5 10 15 <210> 225 <211> 15 <212> PRT <213> Homo sapiens <400> 225 Val His Val Thr Val Pro Gln Glu Leu Phe Glu Trp Val Arg Arg 1 5 10 15 <210> 226 <211> 15 <212> PRT <213> Homo sapiens <400> 226 Val His Val Thr Val Pro Gln Glu Leu Tyr Glu Trp Val Arg Arg 1 5 10 15 <210> 227 <211> 15 <212> PRT <213> Homo sapiens <400> 227 Val His Val Thr Val Pro Gln Glu Leu Trp Glu Trp Val Glu Glu 1 5 10 15 <210> 228 <211> 15 <212> PRT <213> Homo sapiens <400> 228 Val His Val Thr Val Pro Gln Glu Leu Phe Glu Trp Val Glu Glu 1 5 10 15 <210> 229 <211> 15 <212> PRT <213> Homo sapiens <400> 229 Val His Val Thr Val Pro Gln Glu Leu Tyr Glu Trp Val Glu Glu 1 5 10 15 <210> 230 <211> 15 <212> PRT <213> Homo sapiens <400> 230 Val His Val Ser Ile Pro Gln Glu Leu Trp Glu Trp Val Arg Arg 1 5 10 15 <210> 231 <211> 15 <212> PRT <213> Homo sapiens <400> 231 Val His Val Ser Val Pro Gln Glu Leu Trp Glu Trp Val Arg Arg 1 5 10 15 <210> 232 <211> 15 <212> PRT <213> Homo sapiens <400> 232 Val His Val Ser Val Pro Gln Glu Leu Tyr Glu Trp Val Arg Arg 1 5 10 15 <210> 233 <211> 15 <212> PRT <213> Homo sapiens <400> 233 Val Arg Val Thr Ile Pro Gln Glu Leu Trp Glu Trp Val Arg Arg 1 5 10 15 <210> 234 <211> 15 <212> PRT <213> Homo sapiens <400> 234 Val Arg Val Thr Val Pro Gln Glu Leu Tyr Glu Trp Val Arg Arg 1 5 10 15 <210> 235 <211> 15 <212> PRT <213> Homo sapiens <400> 235 Val Arg Val Thr Val Pro Gln Glu Leu Trp Glu Trp Val Arg Arg 1 5 10 15 <210> 236 <211> 15 <212> PRT <213> Homo sapiens <400> 236 Val His Val Thr Val Pro Gln Glu Ile Tyr Glu Trp Val Arg Arg 1 5 10 15 <210> 237 <211> 15 <212> PRT <213> Homo sapiens <400> 237 Val His Val Thr Ile Pro Gln Glu Ile Trp Glu Trp Val Arg Arg 1 5 10 15 <210> 238 <211> 15 <212> PRT <213> Homo sapiens <400> 238 Val His Phe Thr Val Pro Gln Glu Leu Tyr Glu Trp Val Arg Arg 1 5 10 15 <210> 239 <211> 15 <212> PRT <213> Homo sapiens <400> 239 Val Lys Ile Ser Val Pro Ala Asp Leu Trp Asp Trp Ile Asn Lys 1 5 10 15 <210> 240 <211> 15 <212> PRT <213> Homo sapiens <400> 240 Leu Arg Ile Ser Val Pro Ala Asp Leu Trp Asp Trp Ile Asn Lys 1 5 10 15 <210> 241 <211> 15 <212> PRT <213> Homo sapiens <400> 241 Leu Arg Ile Tyr Val Pro Ala Asp Leu Trp Asp Trp Ile Asn Lys 1 5 10 15 <210> 242 <211> 15 <212> PRT <213> Homo sapiens <400> 242 Val Arg Gly Tyr Val Pro Ala Asp Leu Trp Asp Trp Ile Asn Lys 1 5 10 15 <210> 243 <211> 15 <212> PRT <213> Homo sapiens <400> 243 Val Arg Ala Tyr Val Pro Ala Asp Leu Trp Asp Trp Ile Asn Lys 1 5 10 15 <210> 244 <211> 15 <212> PRT <213> Homo sapiens <400> 244 Val Arg Leu Tyr Val Pro Ala Asp Leu Trp Asp Trp Ile Asn Lys 1 5 10 15 <210> 245 <211> 15 <212> PRT <213> Homo sapiens <400> 245 Val Arg Ile Tyr Leu Pro Ala Asp Leu Trp Asp Trp Ile Asn Lys 1 5 10 15 <210> 246 <211> 15 <212> PRT <213> Homo sapiens <400> 246 Ile His Val Thr Ile Pro Leu Glu Ile Phe Glu Trp Leu Gln His 1 5 10 15 <210> 247 <211> 15 <212> PRT <213> Homo sapiens <400> 247 Ile His Val Thr Ile Pro Leu Glu Ile Phe Glu Trp Ala Gln His 1 5 10 15 <210> 248 <211> 15 <212> PRT <213> Homo sapiens <400> 248 Ile His Val Thr Ile Pro Leu Glu Ile Phe Glu Trp Leu Gln Arg 1 5 10 15 <210> 249 <211> 15 <212> PRT <213> Homo sapiens <400> 249 Ile His Val Thr Ile Pro Leu Glu Val Phe Glu Trp Leu Gln His 1 5 10 15 <210> 250 <211> 15 <212> PRT <213> Homo sapiens <400> 250 Ile His Val Thr Ile Pro Gly Glu Ile Phe Glu Trp Leu Gln His 1 5 10 15 <210> 251 <211> 15 <212> PRT <213> Homo sapiens <400> 251 Val Arg Phe Ser Val Pro Gln Glu Ile Tyr Glu Trp Val Arg Arg 1 5 10 15 <210> 252 <211> 15 <212> PRT <213> Homo sapiens <400> 252 Leu Arg Ile Ser Val Pro Leu Glu Ile Phe Glu Trp Leu Gln His 1 5 10 15 <210> 253 <211> 15 <212> PRT <213> Homo sapiens <400> 253 Leu Arg Gly Ser Val Pro Leu Glu Ile Phe Glu Trp Leu Gln His 1 5 10 15 <210> 254 <211> 15 <212> PRT <213> Homo sapiens <400> 254 Val Lys Ile Ser Val Pro Leu Glu Ile Phe Glu Trp Leu Gln His 1 5 10 15 <210> 255 <211> 15 <212> PRT <213> Homo sapiens <400> 255 Val Glu Phe Asn Phe Pro Gln Gln Val Tyr Glu Trp Phe Asp Asp 1 5 10 15 <210> 256 <211> 15 <212> PRT <213> Homo sapiens <400> 256 Val Glu Phe Asn Phe Pro Gln Gln Val Tyr Glu Trp Val Arg Arg 1 5 10 15 <210> 257 <211> 15 <212> PRT <213> Homo sapiens <400> 257 Thr Trp Tyr Val Phe Asn Glu Gln His Gln Glu Tyr Val Arg Lys 1 5 10 15

Claims

1. A peptide capable of inhibiting the binding of IL-17A to ILRA, wherein the amino acid sequence of the peptide is shown as any one of the following: SEQ ID N. 2, SEQ ID N. 3, SEQ ID N. 4, SEQ ID N. 5, SEQ ID N. 6, SEQ ID N. 7, SEQ ID N. 8, SEQ ID N. 9, SEQ ID N. 10, SEQ ID N. 11, SEQ ID N. 12, SEQ ID N. 13, SEQ ID N. 14, SEQ ID N. 15, SEQ ID N. 16, SEQ ID N. 17, SEQ ID N. 18, SEQ ID N.42, SEQ ID N. 111, SEQ ID N. 112, SEQ ID N. 113, SEQ ID N. 114, SEQ ID N. 115, SEQID N. 116, SEQ ID N. 117, SEQ ID N. 118, SEQ ID N. 119, SEQ ID N. 120, SEQ ID N.121, SEQ ID N. 122, SEQ ID N. 123, SEQ ID N. 124, SEQ ID N. 125, SEQ ID N. 126, SEQID N. 127, SEQ ID N. 128, SEQ ID N. 129, SEQ ID N. 130, SEQ ID N. 131, SEQ ID N.132, SEQ ID N. 133, SEQ ID N. 134, SEQ ID N. 135, SEQ ID N. 136, SEQ ID N. 137, SEQID N. 138, SEQ ID N. 139, SEQ ID N. 140, SEQ ID N. 141, SEQ ID N. 142, SEQ ID N.143, SEQ ID N. 144, SEQ ID N. 145, SEQ ID N. 146, SEQ ID N. 147, SEQ ID N. 148, SEQID N. 149, SEQ ID N. 150, SEQ ID N. 151, SEQ ID N. 152, SEQ ID N. 153, SEQ ID N.154, SEQ ID N. 155, SEQ ID N. 158, SEQ ID N. 159, SEQ ID N. 160, SEQ ID N. 161, SEQID N. 162, SEQ ID N. 163, SEQ ID N. 164, SEQ ID N.165、SEQ ID N. 166、SEQ ID N.167、SEQ ID N. 168、SEQ ID N. 169、SEQ ID N. 170、SEQ ID N. 171、SEQ ID N. 172、SEQID N. 173、SEQ ID N. 174、SEQ ID N. 175、SEQ ID N. 176、SEQ ID N. 177、SEQ ID N.178、SEQ ID N. 179、SEQ ID N. 180、SEQ ID N. 181、SEQ ID N. 182、SEQ ID N. 183、SEQID N. 184、SEQ ID N. 185、SEQ ID N. 186、SEQ ID N. 187、SEQ ID N. 188、SEQ ID N.189、SEQ ID N. 190、SEQ ID N. 191、SEQ ID N. 192、SEQ ID N. 193、SEQ ID N. 194、SEQID N. 195、SEQ ID N. 196、SEQ ID N. 197、SEQ ID N. 198、SEQ ID N. 200、SEQ ID N.201、SEQ ID N. 202、SEQ ID N. 203、SEQ ID N. 204、SEQ ID N. 205、SEQ ID N. 206、SEQID N. 207、SEQ ID N. 208、SEQ ID N. 209、SEQ ID N. 210、SEQ ID N. 211、SEQ ID N.212、SEQ ID N. 213、SEQ ID N. 214、SEQ ID N. 215、SEQ ID N. 216、SEQ ID N. 217、SEQID N. 218、SEQ ID N. 219、SEQ ID N. 220、SEQ ID N. 221、SEQ ID N. 222、SEQ ID N.223、SEQ ID N. 224、SEQ ID N. 225、SEQ ID N. 226、SEQ ID N. 227、SEQ ID N. 228、SEQID N. 229、SEQ ID N. 230、SEQ ID N. 231、SEQ ID N. 232、SEQ ID N. 233、SEQ IDN.234、SEQ ID N. 235、SEQ ID N. 236、SEQ ID N. 237、SEQ ID N.238、SEQ ID N. 239、SEQ ID N. 240、SEQ ID N.241、SEQ ID N. 242、SEQ ID N. 243、SEQ ID N. 244、SEQ IDN. 245、SEQ ID N. 246、SEQ ID N. 247、SEQ ID N. 248、SEQ ID N. 249、SEQ ID N. 250、SEQ ID N. 251、SEQ ID N. 252、SEQ ID N. 253、SEQ ID N. 254、SEQ ID N. 255、SEQ IDN. 256、SEQ ID N. 257。.

2. A peptide, wherein the amino acid sequence of the peptide is as shown in any one of the following: SEQ ID N. 21, SEQ ID N.22, SEQ ID N. 23, SEQ ID N. 24, SEQ ID N. 25, SEQ ID N. 26, SEQ ID N. 27, SEQ ID N.28, SEQ ID N. 31, SEQ ID N. 32, SEQ ID N. 33, SEQ ID N. 34, SEQ ID N. 35, SEQ ID N. 36, SEQ ID N 37, SEQ ID N 38, SEQ ID N 39, SEQ ID N 40.

3. A peptide, wherein the amino acid sequence of the peptide is shown in any one of the following: SEQ ID N.19, SEQ ID N.20, SEQ ID N.29, SEQ ID N.30, SEQ ID N.

43.

4. The peptide of any one of claims 1 to 3, which is bound to a protective cap group at the C-terminus and / or N-terminus, wherein the protective cap group bound to the C-terminus is selected from amides, aldehydes, esters, p-nitroaniline, 7-amino-4-methylcoumarin, and the protective cap group bound to the N-terminus is selected from acetyl, formyl, pyroglutamyl, fatty acids, urea, carbamate sulfonamides, alkylamines.

5. The peptide of claim 1, further consisting of SEQ ID N.

2.

6. A bioconjugate comprising the peptide of any one of claims 1 to 5 and a biomolecule, wherein the molecule is bound to the N-terminus and / or C-terminus of the peptide.

7. The bioconjugate of claim 6, wherein the biomolecule is selected from the group consisting of capric acid, hexanoic acid, ascorbic acid, NAG-NAM, NAG, NAM, hyaluronic acid, alginic acid, chitin, (GalNAc)2, Gal-α1, 3-GalNAc and trigalacturonic acid.

8. A pharmaceutical composition comprising the peptide of any one of claims 1 to 5, or the bioconjugate of claim 6 or 7, and at least one pharmaceutically acceptable excipient.

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