Methods of treating dry eye disease

Abstract

The present invention provides methods of treating dry eye and slowing, inhibiting, or delaying the progression of dry eye in a subject. The methods comprise administering a composition comprising a therapeutically effective amount of a compound that inhibits binding of an inflammatory interleukin 17 (IL-17) cytokine to the IL-17 receptor. In embodiments, the compound is a protein or peptide.

Description

[0001] Reference to a sequence listing, table or computer program

[0002] This application contains a sequence listing that has been submitted electronically in XML format. The sequence listing XML is incorporated herein by reference. The XML file was created on June 7, 2023, with the file name 113665-0038-8001WO00_SEQ.xml and a size of 186,910 bytes. Technical Field

[0003] The subject matter described herein relates to methods of treating dry eye disease and slowing, inhibiting or delaying the progression of dry eye disease in a subject by administering a composition comprising a therapeutically effective amount of a compound that inhibits the binding of the inflammatory interleukin 17 (IL-17) cytokine to the IL-17 receptor.

[0004] background

[0005] Dry eye is a multifactorial, inflammatory, immune-mediated ocular surface disease that causes symptoms of discomfort, visual disturbances and tear film instability and ocular surface epithelial cell damage. The normal tear film is a relatively stable film, composed of a superficial lipid layer and an aqueous layer mixed with a mucin gel layer, which is partially adhered to the corneal and conjunctival surface epithelium. The natural tear film is important for lubrication and maintenance of the refractive surface of the eye. Dry eye is a complex disease characterized by dysfunction of one or more components of the tear film, resulting in loss of tear film homeostasis or stability, hyperosmotic shifts in tear film osmotic balance and / or insufficient ocular surface fluid volume. It is characterized by rapid tear film rupture and many symptoms, including dry eyes, eye pain, burning / stinging, foreign body sensation, itching and photophobia.

[0006] Brief Overview

[0007] The following aspects and embodiments thereof described and illustrated below are intended to be exemplary and illustrative, not limiting in scope.

[0008] In one aspect, a method of treating dry eye in a subject diagnosed with dry eye is provided. The method comprises administering directly to the eye of a subject in need thereof a composition comprising a therapeutically effective amount of a compound that inhibits the binding of an inflammatory interleukin 17 (IL-17) cytokine to an IL-17 receptor.

[0009] In another aspect, a method of slowing, inhibiting or delaying the progression of dry eye disease in a subject is provided. The method comprises administering directly to the eye of a subject in need thereof a composition comprising a therapeutically effective amount of a compound that inhibits the binding of an inflammatory interleukin 17 (IL-17) cytokine to an IL-17 receptor.

[0010] In another aspect, a method of reversing the progression of dry eye disease in a subject is provided. The method comprises administering directly to the eye of a subject in need thereof a composition comprising a therapeutically effective amount of a compound that inhibits the binding of an inflammatory interleukin 17 (IL-17) cytokine to an IL-17 receptor.

[0011] In another aspect, a method for restoring tear film homeostasis on the surface of the eye is provided. The method comprises administering directly to the eye of a subject in need thereof a composition comprising a therapeutically effective amount of a compound that inhibits the binding of an inflammatory interleukin 17 (IL-17) cytokine to an IL-17 receptor.

[0012] In one embodiment, the method comprises identifying a subject suffering from mild or moderate dry eye.

[0013] In one embodiment, the method comprises identifying a subject having mild, moderate, or severe dry eye.

[0014] In one embodiment, identifying the subject comprises the use of a questionnaire.

[0015] In one embodiment, the questionnaire is selected from the group consisting of the Visual Analog Scale (VAS), the Ocular Surface Disease Index (OSDI) questionnaire, the Symptom Assessment of Dry Eye (SANDE) questionnaire, the Dry Eye Questionnaire (DEQ), and the Standard Patient Evaluation of Dry Eye (SPEED) questionnaire.

[0016] In one embodiment, the method comprises identifying a subject with mild or moderate dry eye disease and monitoring the progression of the dry eye disease.

[0017] In an embodiment, the progression of dry eye disease is monitored using a dry eye visual analog scale (eg, VAS or SANDE).

[0018] In one embodiment, the composition administered comprises a compound selected from a biomolecule or an organic synthetic compound.

[0019] In one embodiment, the organic synthetic compound is a spirocyclic indane compound or a spirocyclic oxoindoline compound.

[0020] In one embodiment, the biological compound is an antibody having binding affinity for the IL-17 cytokine or an antibody having binding affinity for the IL-17 receptor.

[0021] In one embodiment, the antibody is a monoclonal antibody, a polyclonal antibody, a single chain antibody, a humanized antibody, a recombinant antibody, a chimeric antibody, or an antibody fragment.

[0022] In one embodiment, the antibody is selected from the group consisting of afasevikumab, bimekizumab, brodalumab, ixekizumab, izokibep netakimab, perakizumab, secukinumab, sonelokimab, tibulizumab, vunakizumab, ABY-035, CJM-112, CNTO-6785, DC-806 / S-011806, FPP-003, GR-1501, HB-0017, IMU-035, LZM-012, QX-002N, BH-1657, HB-0043, HT-0017, ILCT-1001, IQ-001, LEO 153339 / LP0200, LP-0200, MT-6194, MYMD-1, ND-016, SCT-650A, SM-17, YBL-004, ABM-60, ETI-1023, LQ-025, LQ -026, ABBV-257, AFB-035, ANB-004, BCD-121, COVA-322, CYT-017-IL17Qb, DLX-2882, DLC-2907, DLX-2909, DLX -3003, E-34935, E-35018, E-35762, E-36041, EBI-006, HEISCO-III-002, IL-17-RC, MEDI-571, MOR-106, MP-02 30. PRS-190, SCH-900117, Y-320, ABT-122, BITS-7201A, CDP-435, EBI-028, IL-17E, JNJ-6118104, KHK-4827 and RG 7624.

[0023] In one embodiment, the biological compound is a protein or peptide that specifically inhibits the binding of IL-17A to the interleukin 17A receptor.

[0024] In one embodiment, the peptide consists of a continuous sequence of 10-25, 12-20, 13-18, 13-16, 14-16 or 15 amino acid residues, and the continuous sequence has at least about 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% sequence identity with the sequence of Formula I. In another embodiment, the peptide consists of a continuous sequence of 12-18 amino acid residues, and the continuous sequence has at least about 70% sequence identity with SEQ ID NO: 1. In another embodiment, the peptide consists of a continuous sequence of 12-18 amino acid residues, and the continuous sequence has at least about 70% sequence identity with any of the sequences of SEQ ID NO: 1-166 identified herein. In another embodiment, the peptide consists of a continuous sequence of 12-18 amino acid residues, and the continuous sequence has at least about 70% sequence identity with any of the sequences of SEQ ID NO: 1-214 identified herein. In another embodiment, the peptide consists of a contiguous sequence of 12-18 amino acid residues having at least about 70% sequence identity to any one of SEQ ID NOs: 1-166 identified herein, with the proviso that the peptide is not SEQ ID NOs: 167-214.

[0025] In one embodiment, the peptide comprises the amino acid sequence of Formula I:

[0026] X1-X2-X3-X4X5-X6-X7-X8-X9-X 10 -X1I-X 12 -X 13 -X 14 -X 15 Formula (I),

[0027] in

[0028] X1 is I, V or L;

[0029] X2 is H, M, R, K or E;

[0030] X3 is V, F or I;

[0031] X4 is T, Q, S, Y or N;

[0032] X5 is I, F, or V;

[0033] X6 is P or G;

[0034] X7 is A, Q or L;

[0035] X8 is D, E or Q;

[0036] X9 is L, W, F, V, or I;

[0037] X 10 is W, Y or F;

[0038] X 11 is D, E or N;

[0039] X 12 is W or F;

[0040] X 13 is I, V, F, or L;

[0041] X 14 is N, R, Q or E; and

[0042] X 15 It is K, R, H or E.

[0043] In an embodiment, the sequence is not IHVTIPADLWDWINK (SEQ ID NO: 167).

[0044] In one embodiment, the amino acid sequence of Formula I is wherein

[0045] (a) X1 is I or V,

[0046] X2 is H, M or R,

[0047] X3 is V or F;

[0048] X4 is T or Q;

[0049] X5 is I, F, or V;

[0050] X6 is P or G;

[0051] X7 is A or Q;

[0052] X8 is D or E;

[0053] X9 is L;

[0054] X 10 is W or Y;

[0055] X is D or E;

[0056] X 12 It is W;

[0057] X 13 is I or V;

[0058] X 14 is N, R or E; and

[0059] X 15 is K, R or E;

[0060] or

[0061] (b) X1 is I or V;

[0062] X2 is H or M,

[0063] X3 is V;

[0064] X4 is T;

[0065] X5 is I;

[0066] X6 is P;

[0067] X7 is A;

[0068] X8 is D;

[0069] X9 is L, W, F, V, or I;

[0070] X 10 is W or Y;

[0071] X 11 is D or E;

[0072] X 12 It is W;

[0073] X 13 is I or V;

[0074] X 14 is N, R or E; and

[0075] X 15 It is K, R or E.

[0076] In an embodiment, the sequence is not IHVTIPADLWDWINK (SEQ ID NO: 167), HVTIPADLWDWIN (SEQ ID NO: 168), IHVTIPADLWDWI (SEQ ID NO: 169) or IHVTIPADLWDW (SEQ ID NO: 170).

[0077] In one embodiment, the peptide 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 the group consisting of amides, aldehydes, esters, p-nitroaniline, 7-amino-4-methylcoumarin. In one embodiment, the protective group cap bound to the N-terminus is selected from the group consisting of acetyl, formyl, pyroglutamyl, fatty acids, urea, carbamate sulfonamides and alkylamines.

[0078] In one embodiment, the peptide is in the form of a dimer formed by two peptides, each of which is a peptide of formula I.

[0079] In one embodiment, the two peptides in the dimer are linked by a polyethylene spacer.

[0080] In one embodiment, the peptide is in the form of a bioconjugate comprising the peptide of Formula I and a biomolecule, wherein the biomolecule is bound to the N-terminus and / or C-terminus of the peptide.

[0081] In one embodiment, the biomolecule is selected from the group consisting of decanoic acid, hexanoic acid, ascorbic acid, NAG-NAM, NAG, NAM, hyaluronic acid, alginic acid, chitin, (GalNAc)2, Gal-α1,3-GalNAc and trigalacturonic acid.

[0082] In one embodiment, the peptide is in a pharmaceutically acceptable composition comprising at least one pharmaceutically acceptable excipient.

[0083] In one embodiment, the administering comprises instillation onto the ocular surface.

[0084] In one embodiment, the administering comprises instillation into the conjunctival sac.

[0085] In one embodiment, the administering comprises administering once daily.

[0086] In one embodiment, the therapeutically effective dose of the peptide is about 0.01-1,000 μM, or about 0.05-750 μM, or about 0.05-500 μM, or at least about 500 μM.

[0087] In one embodiment, the dose is administered once daily.

[0088] In one embodiment, administration is continued for a period of at least about 3 weeks, wherein signs and / or symptoms of dry eye resolve during the disease-free period of the third week of administration. In another embodiment, administration is continued for a period of at least about 3 weeks, wherein signs of dry eye resolve during the disease-free period of the third week of administration.

[0089] In another aspect, a method of restoring goblet cell function and / or density in an eye with signs or symptoms of dry eye disease and / or a method of preventing goblet cell loss in an eye of a person suffering from dry eye disease is provided. The method comprises topically administering to an eye of a subject in need thereof a therapeutically effective amount of a compound that inhibits the binding of IL-17A to an interleukin 17A receptor.

[0090] In another aspect, a method for improving the balance of lacrimal gland, Meibomian gland and conjunctival goblet cell function to restore tear film homeostasis in a subject is provided. The method comprises topically administering to the eye of a subject in need thereof a therapeutically effective amount of a compound that inhibits the binding of IL-17A to an interleukin 17A receptor.

[0091] In another aspect, a method of increasing basal tear production in a human subject is provided. The method comprises topically administering to an eye of a subject in need thereof a therapeutically effective amount of a compound that inhibits the binding of IL-17A to an interleukin 17A receptor.

[0092] In one embodiment, the subject is a subject who is at least about 50 years old (an "elderly" subject). In embodiments, the elderly subject has hormonal changes and / or is over 50, 55, or 60 years old.

[0093] In one embodiment, the compound used in any of the methods is not any one or more of DLSAVCWAFPWDPECH (SEQ ID NO: 171), DSSAVCWAFPHHPLCHMKAT (SEQ ID NO: 172), ADADMCWFFPTSPWCH (SEQ ID NO: 173), DSSAVCWAFPYLPECH (SEQ ID NO: 174), DISAVCWAFPFDPECH (SEQ ID NO: 175), AYECPRLEYDMFGALHCLPS (SEQ ID NO: 176), CPRLEYDMFGALHCL (SEQ ID NO: 177), CLDLQYDPWGALHCI (SEQ ID NO: 178), CFDLQYDPWGALHCI (SEQ ID NO: 179), CLDLQYDMFGALHCV (SEQ ID NO: 180), CLDLVYDPWGALHCI (SEQ ID NO: 181), CWVLEYDMFGALHCR (SEQ ID NO: 182), NO:182), CWALEYDMFGYLHCR (SEQ ID NO:183), CWVLEYDMFGFLHCR (SEQ ID NO:184), CWVLEYDMFGYLHCR (SEQ ID NO:185) and GPYYFDSSGYLYYYYGLDV (SEQ ID NO:186).

[0094] In addition to the exemplary aspects and embodiments described above, further aspects and embodiments will become apparent by reference to the drawings and by study of the following descriptions.

[0095] Additional embodiments of the present method and the like will be apparent from the following description, drawings, examples, and claims. It will be understood from the foregoing and following description that each and every feature described herein, and each combination of two or more such features, is included within the scope of the present disclosure, provided that the features included in such a combination are not mutually inconsistent. In addition, any feature or combination of features may be specifically excluded from any embodiment of the present disclosure. Additional aspects and advantages of the present disclosure are set forth in the following description and claims, particularly when considered in conjunction with the accompanying examples and drawings.

[0096] BRIEF DESCRIPTION OF THE DRAWINGS

[0097] Figure 1 The figure is a graph of the relationship between the score of corneal fluorescein staining (CFS) using the National Eye Institute Industry Workshop scale (0-15 points) and time (days) in mice placed in a controlled environment room (relative humidity: <20%, dry airflow: 15 L / min, temperature: 21-23°C) for 14 days to induce dry eye disease. Corneal epithelial disease was evaluated using corneal fluorescein staining (CFS) and scored using the National Eye Institute Industry Workshop scale (0-15 points) on day 0 (normal baseline), day 4 (before topical treatment instillation), day 7, day 10 and day 14 in mice treated with anti-IL-17A antibody once daily (solid circles) or twice daily (solid squares), lifitegrast Twice a day (inverted triangle), cyclosporine Treatment was performed by topical application to the eye twice daily (triangles) or saline twice daily (diamonds).

[0098] Figure 2A-2B is a bar graph that displays the Figure 1 and Example 1, the indicated treatments improved the immunomodulatory function of Treg cells (as measured by FoxP3 expression) ( Figure 2A ) and inhibiting the differentiation of naive T cells into pathogenic Th17 cells ( Figure 2B ) effect.

[0099] Figures 3A-3E is an image of conjunctival tissue showing the frequency of goblet cells (large black dots in the top cell layer) from the following mice: Healthy mice without dry eye disease ( Figure 3A ) and mice with dry eye (dry eye was induced by placing the mice in a controlled environment chamber) in which saline ( Figure 3B ), Cyclosporine ophthalmic emulsion ( Figure 3C ), lifitegrast ophthalmic solution ( Figure 3D ) or anti-IL-17A antibody ( Figure 3E ) to treat the mice suffering from dry eye disease.

[0100] Figure 3F is a bar graph of the percentage of goblet cells in normal (healthy) mice and mice with dry eye disease (dry eye disease was induced by placing these mice in a controlled environment chamber) treated with saline, cyclosporine, lifitegrast, or an IL-17A inhibitor.

[0101] Figures 4A-4C Human corneal epithelial cells ( Figure 4A ) and images of human corneal epithelial cells co-cultured with Th17 cells, where the latter were treated with saline vehicle ( Figure 4B ) or IL-17 inhibitors (IL-17A antibodies) ( Figure 4C )deal with.

[0102] Figures 5A-5F Human corneal epithelial cells ( Figure 5A ) and bright-light microscopy (40x) of human corneal epithelial cells co-cultured with Th17 cells, the latter of which were treated with saline vehicle ( Figure 5B ) or a dose of 0.1 μM ( Figure 5C )、1μM( Figure 5D )、10μM( Figure 5E ) and 100 μM ( Fig. 5F ) was treated with an IL-17 inhibitor (SEQ ID NO: 1 (VHVTIPADLWDWINK)).

[0103] Fig. 6A Graph of corneal fluorescein staining (CFS) scores using the National Eye Institute Industry Workshop scale (0-15 points) at day 0 (normal baseline), day 4 (before topical treatment instillation), and during treatment: IL-17A antibody (inverted triangles) at a dose of 1% of the IL-17 inhibitor, or IL-17 inhibitor (SEQ ID NO: 1 (VHVTIPADLWDWINK)) at a dose of 0.05 wt % (circles) or 0.1 wt % (diamonds) dissolved in phosphate-buffered saline, administered twice daily on days 4-12; control group administered saline vehicle (squares) twice daily on days 4-12.

[0104] Figure 6B It comes from Fig. 6ABar graph of the percentage of Th17 cells in the draining lymph nodes of the same mice, including mice without dry eye and mice with dry eye (dry eye was induced by placing these mice in a controlled environment chamber), and the mice with dry eye were treated with saline (control) or IL-17A antibody at a dose of 1 wt% (10 mg / mL) or IL-17 inhibitor (SEQ ID NO: 1 (VHVTIPADLWDWINK)) at a dose of 0.05 wt% or 0.1 wt% dissolved in phosphate buffered saline, twice daily for 9 days.

[0105] Figure 7 Graph of the score for corneal fluorescein staining (CFS) in mice using the National Eye Institute Industry Workshop scale (0-15 points) versus time (days) during the treatment phase of a study in mice with dry eye disease, treated twice daily with saline vehicle (open squares) or with an IL-17 inhibitor (SEQ ID NO: 1 (VHVTIPADLWDWINK)) at concentrations of 50 μM (administered twice daily, inverted triangles), 250 μM (administered to separate groups once daily (diamonds) and twice daily (closed squares)), and 500 μM (administered once daily, circles) in phosphate-buffered saline.

[0106] Figure 8 Graph of the relationship between the score of corneal fluorescein staining (CFS) of mice using the National Eye Institute Industry Workshop Scale (0-15 points) after dry eye induction and during treatment in three groups of mice, treated with vehicle (control, open squares) or with IL-17 inhibitor (SEQ ID NO: 1 (VHVTIPADLWDWINK)) at a concentration of 250 μM in phosphate-buffered saline, applied twice daily for 7 days (triangles) or 21 days (squares). DETAILED DESCRIPTION OF THE INVENTION

[0108] I. definition

[0109] Various aspects will now be described more fully below. However, such aspects may be embodied in many different forms and should not be construed as limited to the embodiments described herein; on the contrary, these embodiments are provided to make this disclosure comprehensive and complete and to fully convey its scope to those skilled in the art.

[0110] For convenience, certain terms employed in the specification, examples, and claims are collected here. Unless otherwise defined, all technical and scientific terms used in this disclosure have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs.

[0111] Where a numerical range is provided, it is intended that each intervening value between the upper and lower limits of the range and any other stated value or intervening value within the stated range is encompassed in the present disclosure. For example, if a range of 1 μm to 8 μm is stated, 2 μm, 3 μm, 4 μm, 5 μm, 6 μm and 7 μm are also expressly disclosed, as well as a range of values ​​greater than or equal to 1 μm and a range of values ​​less than or equal to 8 μm.

[0112] The singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "a "polymer" includes a single polymer and two or more polymers, whether the same or different, reference to "an "excipient" includes a single excipient and two or more excipients, whether the same or different, and so forth.

[0113] The term "about", particularly when referring to a given amount, is intended to encompass a deviation of plus or minus 5%.

[0114] "Amino acid" refers to all naturally occurring L-amino acids and includes D-amino acids. Amino acids are identified by either single-letter or three-letter symbols.

[0115] The term "amino acid sequence variant" refers to a molecule having some differences in its amino acid sequence compared to a sequence according to the present disclosure. Amino acid sequence variants of a polypeptide according to the present disclosure (e.g., of a specified sequence) retain the ability to bind to a specific IL-17 cytokine or receptor, such as human IL-17A, or, for example, inhibit the binding of IL-17A to its receptor. Substitution variants are those in which at least one amino acid residue is removed from the same position of a polypeptide according to the present disclosure (e.g., of a specified sequence) and a different amino acid is inserted in its place. These substitutions may be single, in which only one amino acid is substituted in the molecule, or multiple, i.e., two or more amino acids are substituted in the same molecule. Insertion variants are those in which one or more amino acids are inserted in the immediate vicinity of an amino acid at a specific position of a polypeptide (e.g., of a specified sequence). The immediate vicinity of an amino acid refers to the α-carboxyl or α-amino functional group attached to the amino acid. Deletion variants are those in which one or more amino acids are removed from a polypeptide of a specified sequence.

[0116] For natural polypeptides and functional derivatives thereof, "identity" is defined as the percentage of amino acid residues in a candidate sequence that are identical to the residues of a corresponding natural polypeptide after aligning the sequences and introducing gaps, if necessary, to achieve maximum percent identity and without considering any conservative substitutions as part of sequence identity. Extensions or insertions at the N-terminus or C-terminus should not be interpreted as reducing identity. Alignment methods and computer programs are well known. Percent identity can be determined by standard alignment algorithms, such as the basic local alignment search tool (BLAST) described by Altshul et al. ((1990) J. Mol. Biol., 215: 403 410); Needleman et al.'s algorithm ((1970) J. Mol. Biol., 48: 444 453); or Meyers et al.'s algorithm ((1988) Comput. Appl. Biosci., 4: 11 17). One set of parameters can be a Blosum 62 scoring matrix, with a gap penalty of 12, a gap extension penalty of 4, and a frameshift gap penalty of 5. The percent identity between two amino acid or nucleotide sequences can also be determined using the algorithm of E. Meyers and W. Miller ((1989) CABIOS, 4: 11-17), which has been incorporated into the ALIGN program (version 2.0), using the PAM120 weight residue table, a gap length penalty of 12, and a gap penalty of 4.

[0117] The term "IL-17" refers to an IL-17 cytokine, such as IL-17A, IL-17B, IL-17C, IL-17D or IL-17E.

[0118] "IL-17A" refers to a cytokine formerly known as CTLA8, and includes wild-type IL-17A, polymorphic variants of IL-17A, and functional equivalents of IL-17A from various species (e.g., humans, mice, and monkeys). Functional equivalents of IL-17A have at least about 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% overall sequence identity with wild-type IL-17A (e.g., human IL-17A), and, in embodiments, functional fragments are functional fragments that substantially retain the ability to induce IL-6 production by human dermal fibroblasts.

[0119] "IL-17 inhibitor" refers to a compound that binds to IL-17 cytokine and / or to an IL-17 receptor (eg, IL-17RA, RB, RC, RD, or RE) to inhibit the interaction of IL-17 cytokine with its receptor.

[0120] "IL-17 cytokine inhibitor" refers to a compound that binds to an IL-17 cytokine to inhibit its interaction with its receptor. In an embodiment, an IL-17 cytokine inhibitor specifically binds to a specific IL-17 cytokine and is referred to according to the cytokine to which it binds, for example, an "IL-17A cytokine inhibitor" refers to a compound that specifically binds to an IL-17A cytokine to inhibit its binding or interaction with an IL-17A receptor, thereby inhibiting the formation of an IL-17A / IL-17RA / IL17RC complex.

[0121] Compounds that "inhibit" one or more functional properties of IL-17 (e.g., biochemical, immunochemical, cellular, physiological or other biological activity, etc.) are determined according to methods known in the art and described herein, and the compound causes a statistically significant reduction in a specific activity relative to that seen in the absence of the compound (or when a control compound of irrelevant specificity is present). Compounds that are IL-17 inhibitors or IL-17 cytokine inhibitors provide a statistically significant reduction, for example, at least 10%, at least 50%, 80% or 90% reduction in the measured parameter, and in certain embodiments may inhibit greater than 95%, 98% or 99% of the functional activity of IL-17.

[0122] The phrase "pharmaceutically acceptable" is used herein to refer to those compounds, salts, compositions, dosage forms, etc., which are suitable (within the scope of reasonable medical judgment) for use in contact with the tissues of humans and / or other mammals without excessive toxicity, irritation, allergic response or other problems or complications, and are matched with a reasonable benefit / risk ratio. In some aspects, "pharmaceutically acceptable" means approved by a U.S. federal or state government regulatory agency, or listed in the U.S. Pharmacopeia or other generally recognized pharmacopeia, for use in mammals (e.g., animals) and more specifically in humans.

[0123] As used herein, the terms "subject" and "patient" include any human or non-human animal. The term "non-human animal" includes all vertebrates, e.g., mammals and non-mammals, such as non-human primates, mice, rabbits, sheep, dogs, cats, horses, cows, chickens, amphibians, reptiles, etc.

[0124] The phrase "substantially identical" means that the relevant amino acid sequence or nucleotide sequence will be identical or have insubstantial differences (e.g., by conservative amino acid substitutions) compared to a specific reference sequence. Insubstantial differences include minor amino acid changes, such as 1 or 2 substitutions in 5 amino acid sequences of a specified region. In the case of antibodies, the second antibody has the same specificity and has at least 50% of the same affinity. Sequences substantially identical to the sequences disclosed herein (e.g., at least about 85% sequence identity) are also part of this application. In some embodiments, the sequence identity can be about 90% or more, for example, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more.

[0125] As used herein, "therapeutically effective amount" refers to an amount of an IL-17 inhibitor that is effective when administered to a subject (e.g., a human patient) in a single dose or multiple doses to slow, inhibit or delay the progression of dry eye disease and / or to treat, regress or cure dry eye disease.

[0126] The terms "treatment" or "treat" refer to curative or disease-correcting treatment in a patient diagnosed with dry eye disease.

[0127] The compositions of the present disclosure may include, consist essentially of, or consist of the disclosed components.

[0128] All percentages, parts and ratios are based upon the total weight of the composition and all measurements made are at about 25°C, unless otherwise specified.

[0129] Less than the entirety of the disclosure may be claimed for any reason by reserving the right to proviso out or exclude at any time any individual member of any such group (including any subrange or combination of subranges within that group) which may be claimed as a range or in any similar manner. In addition, less than the entirety of the disclosure may be claimed for any reason by reserving the right to proviso out or exclude at any time any individual substituent, analog, compound, ligand, structure or group thereof, or any member of a claimed group.

[0130] In the entire content of this disclosure, various patents, patent applications and publications are cited. The entire disclosure of these patents, patent applications and publications is incorporated into this disclosure by reference to more fully describe the prior art known to those skilled in the art as of the date of this disclosure. If there is any inconsistency between the cited patents, patent applications and publications and this disclosure, the disclosure shall prevail.

[0131] II. method

[0132] Methods for treating dry eye disease and methods for slowing, inhibiting or delaying the progression of dry eye disease in a subject are provided. The methods include administering a composition comprising a therapeutically effective amount of a compound that inhibits the binding of the inflammatory interleukin 17 (IL-17) cytokine to the IL-17 receptor, referred to as an IL-17 inhibitor. Studies conducted to support the methods are described with respect to Examples 1-7.

[0133] In the first study described in Example 1, the IL-17 cytokine inhibitor, lifitegrast ophthalmic solution, was compared with and cyclosporine ophthalmic emulsion Treatment of dry eye disease. The IL-17 cytokine inhibitor is an anti-IL-17A antibody formulated as 1 wt% in phosphate-buffered saline. Dry eye disease was induced in mice before treatment, and after the induction of dry eye disease, the mice were divided into several groups for the following treatments: (i) anti-IL-17A antibody, once a day; (ii) 1 wt% anti-IL-17A antibody, twice a day; (iii) 5 wt% lifitegrast, twice a day; (iv) 0.05 wt% cyclosporine, twice a day; or (v) saline as a control, twice a day. The test compound was topically applied to the eyes for 9 days (from the 4th day to the 12th day), and the severity of the disease was assessed using corneal fluorescein staining (CFS) on the 4th, 7th, 10th and 12th days, and scored using the National Eye Institute Industry Workshop Scale (0-15 points).

[0134] Figure 1 is a graph of dry eye scores versus time (days) for subjects with dry eye treated with anti-IL-17A antibody once daily (solid circles) or twice daily (solid squares), lifitegrast ( 5wt%) twice a day (inverted triangle), cyclosporine ( 0.05 wt %) twice daily (triangles) or saline twice daily (diamonds) were topically applied to the eyes to treat the subjects with dry eye. Topical treatment of the eyes with the IL-17 cytokine inhibitor was more effective than lifitegrast or cyclosporine in treating dry eye, as measured by disease scores. Subjects treated with cyclosporine who had dry eye scores of approximately 8 on day 4 of the study experienced little, if any, reduction in disease severity, with their dry eye scores remaining at approximately 8 during treatment. The reduction in disease severity in subjects treated with lifitegrast who had dry eye scores of approximately 8 on day 4 of the study showed no improvement during the first 6 days of treatment, followed by an improvement in disease scores on day 12. Subjects treated with the IL-17 cytokine inhibitor reported an improvement in the severity of dry eye within 3 days of treatment, and the severity of dry eye continued to improve for the remainder of the treatment period.

[0135] Figure 1 The data also showed that the IL-17 cytokine inhibitor was effective in treating dry eye disease when administered once daily. There was no statistical difference in the reduction of disease scores in the test cohorts treated with the IL-17 cytokine inhibitor once daily or twice daily. Figure 2A This finding is also supported by the data shown, where once-daily or twice-daily treatment with an IL-17 cytokine inhibitor restored the presence of protective immunoregulatory T cells (Tregs). Figure 2B showed that treatment with an IL-17 cytokine inhibitor either once a day or twice a day effectively reduced the percentage of pathogenic Th17 immune cells in the draining lymph nodes.

[0136] Therefore, a method for treating dry eye is provided, wherein a subject suffering from dry eye is treated with an IL-17 cytokine inhibitor. In one embodiment, the IL-17 cytokine inhibitor is applied to the eye once a day. In one embodiment, the IL-17 cytokine inhibitor is applied to the eye twice a day.

[0137] Continuing the study of Reference Example 1, eye tissue was obtained from mice and tissue sections were prepared for microscopic analysis. Cross-sections of the conjunctiva were stained and images were examined to quantify goblet cell density. Goblet cells in the conjunctival epithelium secrete mucin to the surface of the eye. Figure 3A is an image of the conjunctiva from a healthy mouse without dry eye disease, and goblet cells are visible as darkly stained fat cells. Figure 3B-3E are images of the conjunctiva from animal subjects with dry eye disease treated twice daily with saline as a control ( Figure 3B ), Cyclosporine ophthalmic emulsion ( Figure 3C ), lifitegrast ophthalmic solution ( Figure 3D ) or anti-IL-17A antibody ( Figure 3E ) treatment. The number of goblet cells was counted, and Figure 3F Is a bar graph of the percentage of normal goblet cells in normal (healthy) mice and mice with dry eye treated with saline, cyclosporine, lifitegrast or IL-17A inhibitor. There was no loss in the number of goblet cells in subjects treated with IL-17A inhibitor, and IL-17A inhibitor prevented goblet cell loss in conjunctival tissue. Subjects with dry eye and treated with cyclosporine or lifitegrast had approximately 60% of the normal number of goblet cells. The preservation and / or restoration of goblet cell function and / or density demonstrates that IL-17A inhibitor is a disease-modifying therapy for dry eye, treating the disease itself rather than just ameliorating symptoms of dry eye, and restoring tear film homeostasis.

[0138] Dry eye disease is characterized by a decrease in tear volume, rapid tear film breakdown, and / or increased evaporative properties of the tear film layers. Tears are composed of three layers: an outer oily or lipid layer, a middle aqueous layer (watery layer), and an inner mucin layer. The production of a healthy tear film involves the meibomian glands, lacrimal glands, and goblet cells. The outer lipid layer of the tear film is produced by the meibomian glands, and the function of this layer is to maintain the tear film on the surface of the eye to prevent it from evaporating too quickly. The lacrimal glands produce the middle aqueous layer, which includes most of the tear volume and tear flow. This layer is an aqueous mixture that contains proteins, enzymes, antibodies, and growth factors that have cell-protective, anti-inflammatory, and antimicrobial effects. The aqueous layer nourishes the cornea and conjunctiva that cover the entire front of the eye and the inside of the eyelids. The innermost mucin layer is produced by goblet cells, and its function is to bind water in the aqueous layer to ensure that the eye remains moist. As Figure 3B and 3F As shown in the data, dry eye is associated with reduced function and / or density of conjunctival goblet cells. Therefore, in one embodiment, a method of restoring goblet cell function and / or density in an eye with signs or symptoms of dry eye is provided. In another embodiment, a method of preventing goblet cell loss in the eye of a person suffering from dry eye is provided. The method comprises topically administering to the eye of a subject in need thereof a therapeutically effective amount of a compound that inhibits the binding of an IL-17 cytokine to an IL-17 receptor. In one embodiment, the compound inhibits the binding of IL-17A to an IL-17A receptor.

[0139] In another embodiment, a method of improving the balance of the functions of the lacrimal glands, meibomian glands, and conjunctival goblet cells to restore tear film homeostasis in a subject is provided. The method comprises topically administering to the eye of a subject in need thereof a therapeutically effective amount of a compound that inhibits the binding of an IL-17 cytokine to an IL-17 receptor. In one embodiment, the compound inhibits the binding of IL-17A to an IL-17A receptor. In an embodiment, the subject in need thereof is a person suffering from dry eye disease.

[0140] In another embodiment, a method of increasing basal tear production in an elderly subject is provided. The method comprises topically administering to the eye of a subject in need thereof a therapeutically effective amount of a compound that inhibits the binding of an IL-17 cytokine to an IL-17 receptor. In one embodiment, the compound inhibits the binding of IL-17A to an IL-17A receptor. In an embodiment, the subject in need thereof is a person suffering from dry eye disease. In one embodiment, the elderly subject has hormonal changes or is over 50 years old.

[0141] In another study described in Example 2, co-cultures of human corneal epithelial cells and Th17 cells were prepared to simulate the in vivo interaction between Th17 cells and corneal tissue in dry eye disease. The therapeutic effect of an IL-17 inhibitor in the form of an antibody, which inhibits the binding of IL-17A to its receptor, was tested to protect human corneal epithelial cells from Th17-induced damage. The density and morphology of human corneal epithelial cells in co-cultures with and without IL-17 inhibitors were observed under a bright light inverted microscope equipped with a camera. Microscopic images of the cells are shown in Figure 2. Figures 4A-4C As shown, Figure 4A The images in the figure are human corneal epithelial cells without Th17 cells or IL-17 treatment as a control. Figure 4B are images of co-cultures of human corneal epithelial cells and Th17 cells treated with saline vehicle, and Figure 4C is an image of a co-culture of human corneal epithelial cells and Th17 cells treated with an IL-17 inhibitor. The IL-17 inhibitor protected human corneal epithelial cells from Th17-mediated corneal damage, which was observed in human corneal epithelial cells ( Figure 4C ) and control cultures of human corneal epithelial cells ( Figure 4A ) have comparable cell density and morphology can be clearly seen.

[0142] Similar studies were performed using co-cultures of human corneal epithelial cells and Th17 cells to test the therapeutic effect of an IL-17A inhibitor in the form of a peptide identified as SEQ ID NO: 1 (VHVTIPADLWDWINK), wherein the IL-17A inhibitor in the form of the peptide inhibits the binding of IL-17A to the IL-17A receptor; to evaluate whether the peptide protects human corneal epithelial cells from Th17-induced damage. As described in Example 3, appropriate concentrations of IL-17 inhibitory peptides were prepared using phosphate-buffered saline to provide compositions having concentrations of 0.1 μM, 1 μM, 10 μM, and 100 μM of IL-17 inhibitors. After 24 hours of co-culture in the presence of each treatment dose of IL-17 inhibitor, the density and morphology of human corneal epithelial cells were observed under a bright light inverted microscope equipped with a camera. Representative images are shown in Figure 3. Figures 5A-5F shown. Figure 5A The images in are images of cultured human corneal epithelial cells without Th17 cells or treated with IL-17 as a control. Figure 5B The images in are images of co-cultures of human corneal epithelial cells and Th17 cells treated with saline vehicle. Figures 5C-5F The images in the figure are images of co-cultures of human corneal epithelial cells and Th17 cells treated with IL-17A inhibitor at concentrations of 0.1 μM, 1 μM, 10 μM, and 100 μM, respectively. After 24 hours, significant death of human corneal epithelial cells was observed in cells treated with vehicle alone ( Figure 5B In contrast, co-cultures treated with all concentrations of the IL-17 inhibitor protected human corneal epithelial cells from Th17-mediated corneal damage ( Figures 5C-5F ), which was compared with the human corneal epithelial cell culture in the control ( Figure 5A ) have comparable cell density and morphology can be clearly seen. Figures 5C-5F The images of human corneal epithelial cells also showed that IL-17 inhibitors were not toxic to human corneal epithelial cells.

[0143] Example 4 describes a study demonstrating that IL-17 inhibitors are effective in slowing, inhibiting and / or delaying the progression of dry eye disease. Four days after dry eye induction, mice were treated with IL-17 inhibitors. The IL-17 inhibitor is an anti-IL-17A antibody at a dose of 1 wt% dissolved in phosphate-buffered saline, or a peptide that inhibits IL-17A binding to the IL-17A receptor identified as SEQ ID NO: 1 (VHVTIPADLWDWINK). The IL-17A peptide inhibitor was formulated in phosphate-buffered saline at doses of 0.05 wt% and 0.1 wt%. After 9 days of treatment with IL-17 inhibitory antibodies or peptides, ocular surface damage was assessed using corneal fluorescein staining (CFS) on days 0 (normal baseline), 4 (before topical treatment instillations), 7, 10, and 12, and scored using the National Eye Institute Industry Workshop Scale (0-15 points).

[0144] The results are shown in Fig. 6A In which the relationship between the score of corneal fluorescein staining (CFS) using the National Eye Institute Industry Workshop Scale (0-15 points) on day 0 (normal baseline) and time is shown. CFS scores were determined on day 4 (before topical treatment instillation) and during treatment with IL-17 inhibitors. The data showed that two doses of IL-17 inhibitory peptide (SEQ ID NO: 1, 0.05wt% (circle) and 0.1wt% (diamond)) and IL-17 inhibitory antibodies (1wt%, inverted triangle) effectively improved the progression of dry eye disease compared to the vehicle-treated cohort (hollow square). In the absence of treatment with IL-17 inhibitors, the CFS score of untreated subjects (hollow squares) on day 4 of the study was close to 8, and the CFS score remained around 8 during the study, indicating that dry eye disease continued to progress without treatment. In subjects treated with IL-17 inhibitors, CFS scores decreased after receiving IL-17 inhibitor treatment, and preparations containing IL-17 inhibitors reduced the severity of dry eye disease and inhibited its progression.

[0145] At the end of treatment, ocular draining lymph nodes were collected and the frequency of IL-17+CD4+Th17 cells in the tissues was determined. Figure 6B Shown are the percentages of Th17 cells in the draining lymph nodes of mice without dry eye disease and mice with dry eye disease and treated with saline vehicle (control) or with an IL-17 inhibitor. The IL-17 inhibitor suppressed pathogenic Th17 cells in ocular tissue.

[0146] Therefore, in one embodiment, a method of slowing, inhibiting or delaying the progression of dry eye disease in a subject is provided. The method comprises administering a composition directly to the eye of a subject in need thereof, the composition comprising a therapeutically effective amount of a compound that inhibits the binding of an inflammatory interleukin 17 (IL-17) cytokine to an IL-17 receptor. In another embodiment, a method of reversing the progression of dry eye disease in a subject is provided, wherein such a compound is administered directly to the eye of a subject in need thereof, the compound inhibiting the binding of an inflammatory interleukin 17 (IL-17) cytokine to an IL-17 receptor.

[0147] Additional studies were performed to further evaluate the IL-17A peptide inhibitor. Example 5 describes a study in which mice with dry eye were treated with an IL-17A peptide inhibitor (SEQ ID NO: 1 (VHVTIPADLWDWINK)) at various doses (50 μM, 250 μM, and 500 μM), administered once or twice daily. Figure 7 Results for the treatment groups are shown for the first 7 days of the treatment period. Figure 7 The data in show that treatment of dry eye with an IL-17A peptide inhibitor at doses of 50 μM twice daily (inverted triangles) and 250 μM once daily (diamonds) resulted in no or only transient improvement relative to treatment with saline twice daily (open squares). Improvements were seen in subjects treated with the IL-17A inhibitor at doses of 250 μM twice daily (closed squares) and 500 μM once daily (circles), as evidenced by a significant reduction in CFS scores.

[0148] In another study, dry eye was induced in mice as described in Example 5. Mice with similar dry eye severity were randomly divided into three treatment groups (n=6) and treated with IL-17A peptide inhibitor (SEQ ID NO: 1) at a dose of 250 μM twice daily after induction of dry eye, one group of mice was treated until day 7 of the study and then used vehicle, while the other group of mice was treated with IL-17 inhibitor for 21 days. Group 3 was a control group treated with saline vehicle for 21 days. Corneal fluorescein staining (CFS) was evaluated on days 3, 7, 10, 14, 17, and 21, and the scores were scored using the National Eye Institute Industry Workshop Scale (0-15 points). The results are shown in Table 1. Figure 8As shown. From the increase in CFS scores in the 21-day study, it was observed that the dry eye disease of the subjects in the vehicle (control) treatment group (square) continued to progress and worsen. Subjects treated with IL-17 inhibitors experienced reduced CFS scores (squares, triangles), indicating that the disease condition has improved. For the treated animals (squares) in the 21-day study, a continuous improvement in the severity of the disease and the inhibition of the progression of the disease were observed, while the subjects (triangles) who were treated for 7 days and then no longer treated showed the recovery of the severity of the disease and the continuous progression of the disease. Treatment of dry eye with IL-17 inhibitors prevents and / or reduces the progression of dry eye.

[0149] In embodiments, a CFS score of about 2-3 using the NEI / industry workshop standard is associated with mild dry eye, about 4-7 is associated with mild dry eye, about 8-10 is associated with moderate dry eye, and about 11-15 is associated with severe dry eye. The initial CFS score of the subject in this study is about 8, or suffers from moderate dry eye. In another embodiment, the CFS score using the VAS scale shows that the score of mild dry eye is about 10-20, the score of mild dry eye is about 30-40, the score of moderate dry eye is about 50-70), and the score of severe dry eye is about 80-100. In reference to Example 5, a study using the NEI / industry workshop scale for CFS scoring was performed, and treatment with an IL-17 inhibitor for 7 days improved dry eye, wherein the CFS score on the 7th day was 4.5-5. In subjects treated with IL-17 inhibitors to day 21, CFS scores continued to decline, with CFS scores on days 18-21 of about 2.5 to 3 (which was associated with treatment, where the disease was mild). However, discontinuation of treatment on day 7 caused the disease to progress from mild (CFS score of 4.5 on day 7) to the upper limit of mild and close to moderate (CFS score of 7 on day 14). Therefore, this study shows that treatment of dry eye with IL-17 inhibitors is effective in preventing, delaying, inhibiting or reducing the progression of the disease. In one embodiment, the treatment is effective in preventing, delaying, inhibiting or reducing the progression of mild dry eye to moderate dry eye. In one embodiment, the treatment is effective in preventing, delaying, inhibiting or reducing the progression of mild dry eye to severe dry eye. In one embodiment, the treatment is effective in preventing, delaying, inhibiting or reducing the progression of moderate dry eye to severe dry eye.

[0150] Figure 8The data in also prove that treating dry eye with IL-17 inhibitors is a disease repair therapy. Subjects without dry eye have an average baseline CFS score of about 2. The treatment of subjects with dry eye with daily administration of IL-17 inhibitors has eliminated the condition and achieved near disease regression. Subjects with dry eye have an initial CFS score of about 8, and within 9 days of treatment with IL-17 inhibitors, the CFS score is close to the score of subjects without dry eye. The results show that the treatment method is a disease repair therapy that achieves near or complete disease regression. In one embodiment, subjects with dry eye are treated at least every day for 2 weeks, 3 weeks, 4 weeks, 2 months, 3 months, 6 months or longer to substantially resolve the disease.

[0151] Methods for restoring tear film homeostasis on the surface of the eye are also contemplated. Homeostasis refers to a relatively stable state of equilibrium between interdependent elements, particularly a condition maintained by physiological processes. With respect to the tear film, it is understood that the tear film composition is dynamic and in a constant state of flux, which responds to environmental conditions to maintain homeostasis on the ocular surface. Traditionally, the tear film has been described as consisting of three separate and distinct layers: a mucin layer, an aqueous layer, and a lipid layer. However, it is now recognized that mixing between the mucin layer and the aqueous layer produces a gradient in which the mucin concentration decreases toward the aqueous layer. This aqueous-mucin layer forms a hydrated gel with complex biology, which is then covered by a lipid layer having its own highly ordered structure. Treatment with an IL-17 inhibitor restores tear film homeostasis, making the tear film of a person with dry eye disease more like that of a person without dry eye disease. Example 6 describes, by way of example, the measurement of tear film homeostasis by tear film breakup time. Tear film homeostasis can also be inferred from CFS scores or from the Schirmer test. Tear film breakup time and ocular status can also be used to assess the balance of lacrimal gland, meibomian gland and conjunctival goblet cell function to restore tear film homeostasis in a subject. Methods for restoring homeostasis of the immunoregulatory function of regulatory T cells in the eye are also contemplated. Dry eye disease is at least in part an immune response, stemming from an imbalance between protective immunoregulation of the ocular surface and Th17-mediated proinflammatory pathways. Chronic dry eye disease involves an inflammatory response cycle that persists due to resistance of pathogenic, IL-17 cytokine-secreting Th17 cells to inhibition of regulatory T cells (Treg). Uncontrolled Th17 activity persists dry eye disease, while blocking IL-17 can reduce ocular surface inflammation, reduce the trafficking of immune cells from the ocular surface to lymphoid tissues, and restore the immunoregulatory function of Treg. IL-17 inhibitors are applied to the ocular area, particularly the ocular surface, to reduce inflammation in the eye by blocking IL-17, a pathogenic inflammatory cytokine of dry eye disease. Blocking IL-17 leads to downregulation of inflammatory cytokines including TNF-α, IL-1, IL-6, IL-8, IL-23, and MMP-9. Blocking IL-17 also restores the immunoregulatory function of regulatory T cells (Tregs), breaking the cycle of dry eye disease. Subsequently, Tregs suppress other non-IL-17-related inflammatory factors, and the trafficking of immune cells from the ocular surface to lymphoid tissues is reduced.

[0152] Composition components

[0153] The methods described herein include administering to the eye a composition comprising a therapeutically effective amount of a compound that inhibits the binding of the inflammatory interleukin 17 (IL-17) cytokine to the IL-17 receptor.Exemplary compounds and other composition components are now described.

[0154] IL-17 is a potent proinflammatory cytokine produced by a new lineage of CD4 T cells (Th17). IL-17 signals through a heteromeric receptor complex composed of IL-17RA and IL-17RC. IL-17 has pleiotropic effects on several immune and non-immune cells, providing a link between T cell activation and inflammatory responses. In addition, IL-17 is additive or synergistic with other proinflammatory cytokines (such as TNFα, IL1β or IL6), leading to the amplification of the inflammatory process. IL-17 (also known as IL-17A) is part of a larger family that includes 6 cytokines, known as IL-17A, IL-17B, IL-17C, IL-17D, IL-17E and IL-17F. All members of this family share a common protein structure. Among these family members, IL-17A and IL-17F are most commonly expressed in immune cells. In the methods described herein, one or more of these family members are targeted by antagonists to inhibit or modify their activity.

[0155] In one embodiment, the IL-17 inhibitor is a synthetic organic or inorganic compound. Macrocyclic compounds that inhibit the interaction of IL-17 cytokines with their receptors include, for example, macrocyclic (Wang et al., MedChemComm, 9 (1): described in 22-26 (2018)) and spirocyclic indane compounds (such as those described in WO20200011731, incorporated herein by reference) or spirocyclic oxoindoles (such as those described in WO2019 / 229079, incorporated herein by reference).

[0156] Embodiments in which the compound that inhibits the binding of the IL-17 cytokine to the IL-17 receptor is a protein or peptide include administering a purified and / or isolated protein or peptide. As used herein, an "isolated" or "purified" polypeptide or protein or peptide is substantially free of other cell materials or culture medium when produced by recombinant technology, or substantially free of chemical precursors or other chemicals when chemically synthesized. The purified compound is at least 60% by weight (dry weight) of the compound of interest. Preferably, the preparation is at least 75% by weight of the compound of interest, more preferably at least 90%, and most preferably at least 99%. Purity can be measured by any appropriate standard method, for example, by column chromatography, polyacrylamide gel electrophoresis or HPLC analysis.

[0157] In an embodiment, the IL-17 inhibitor is a biological compound, such as an antibody or a peptide. For example, antibodies with specific binding to IL-17 cytokines or IL-17 receptors are described in US9,872,901 (incorporated herein by reference). The antibody can be a monoclonal antibody or a polyclonal antibody. The antibody under consideration binds to one or more sequences in IL-17 or IL-17 receptor polypeptides. In some embodiments, the antibody includes a single-chain antibody, a humanized antibody, a recombinant antibody, or a chimeric antibody. In other embodiments, the IL-17 inhibitor is an antibody fragment, including but not limited to Fab, F(ab')2, Fab'Fv, single-chain Fv, etc. The antibody against IL-17 may be a reformulated or humanized derivative of, or binds to an epitope of, a human IL-17 affinity purified polyclonal antibody, a human IL-17 allophycocyanin monoclonal antibody, a human IL-17 biotinylated affinity purified polyclonal antibody, a human IL-17 monoclonal antibody, a human IL-17 monoclonal antibody, a human IL-17 phycoerythrin monoclonal antibody, a mouse IL-17 affinity purified polyclonal antibody, a mouse IL-17 biotinylated affinity purified polyclonal antibody, a mouse IL-17 monoclonal antibody, or a mouse IL-17 monoclonal antibody (each of which can be commercially obtained from, for example, R&D Systems). Preferably, the neutralizing antibody or function blocking antibody against IL-17 may be a reformulated or humanized derivative of, or binds to an epitope of, a monoclonal anti-human IL-17 antibody, an anti-human IL-17 antibody, a polyclonal antibody raised in goats, or a recombinant human IL-17R / Fc chimera. Antibodies to IL-17 receptor (IL-17R) may be de novo or humanized derivatives of, or bind to an epitope of, human IL-17R affinity purified polyclonal antibody, human IL-17R allophycocyanin monoclonal antibody, human IL-17R biotinylated affinity purified polyclonal antibody, human IL-17R fluorescein monoclonal antibody, human IL-17R monoclonal antibody, human IL-17R monoclonal antibody, human IL-17R monoclonal antibody, human IL-17R phycoerythrin monoclonal antibody, mouse IL-17R affinity purified polyclonal antibody, mouse IL-17R biotinylated affinity purified polyclonal antibody, or mouse IL-17R monoclonal antibody (each commercially available from R&D Systems).

[0158] In an embodiment, the IL-17 inhibitor is an antibody selected from secukinumab, ixekizumab, brodalumab, ABT-122, KHK 4827, perakizumab, RG 7624, ANB004 or COVA322. Secukinumab is a fully human IL-17A specific monoclonal antibody derived from the human IgG1κ isotype, and ixekizumab is a humanized IgG4 antibody. Both secukinumab and ixekizumab are specific for IL-17A homodimers and IL-17A. ABT-122 is a dual variable domain immunoglobulin targeting TNF and IL-17A. Brodalumab is a high affinity neutralizing IL-17RA human monoclonal antibody that blocks the biological activity of IL-17, IL-17F, heterodimers composed of 17A / 17F, or 17E. In another embodiment, the antibody is as described in US Pat. No. 9,765,140 (incorporated herein by reference).

[0159] In one embodiment, the IL-17 inhibitor is a peptide capable of inhibiting the binding of IL-17A to IL-17RA. In one embodiment, the peptide binds to IL-17A with high affinity and inhibits its interaction with the ILRA receptor, thereby inhibiting IL-17A signaling. Exemplary peptides are described in WO2020 / 021103 (incorporated herein by reference). In one embodiment, the peptide comprises an amino acid sequence of Formula I:

[0160] X1-X2-X3-X4X5-X6-X7-X8-X9-X 10 -X1I-X 12 -X 13 -X 14 -X 15 Formula (I), wherein

[0161] X1 is I, V or L;

[0162] X2 is H, M, R, K or E;

[0163] X3 is V, F or I;

[0164] X4 is T, Q, S, Y or N;

[0165] X5 is I, F, or V;

[0166] X6 is P or G;

[0167] X7 is A, Q or L;

[0168] X8 is D, E or Q;

[0169] X9 is L, W, F, V, or I;

[0170] X 10 is W, Y or F;

[0171] X 11 is D, E or N;

[0172] X 12 is W or F;

[0173] X 13 is I, V, F, or L;

[0174] X 14 is N, R, Q or E;

[0175] X 15 is K, R, H or E;

[0176] Provided that the sequence is not IHVTIPADLWDWINK (SEQ ID NO: 167). In another embodiment, Formula I is selected from (a) X1 is I or V,

[0177] X2 is H, M or R,

[0178] X3 is V or F;

[0179] X4 is T or Q;

[0180] X5 is I, F, or V;

[0181] X6 is P or G;

[0182] X7 is A or Q;

[0183] X8 is D or E;

[0184] X9 is L;

[0185] X 10 is W or Y;

[0186] X is D or E;

[0187] X 12 It is W;

[0188] X 13 is I or V;

[0189] X 14 is N, R or E;

[0190] X 15 is K, R or E;

[0191] Provided that the sequence is not IHVTIPADLWDWINK (SEQ ID NO: 167); or

[0192] (b) X1 is I or V;

[0193] X2 is H or M,

[0194] X3 is V;

[0195] X4 is T;

[0196] X5 is I;

[0197] X6 is P;

[0198] X7 is A;

[0199] X8 is D;

[0200] X9 is L, W, F, V, or I;

[0201] X 10 is W or Y;

[0202] X 11 is D or E;

[0203] X 12 It is W;

[0204] X 13 is I or V;

[0205] X 14 is N, R or E;

[0206] X 15 is K, R or E;

[0207] The proviso is that the sequence is not IHVTIPADLWDWINK (SEQ ID NO: 167), HVTIPADLWDWIN (SEQ ID NO: 168), IHVTIPADLWDWI (SEQ ID NO: 169) or IHVTIPADLWDW (SEQ ID NO: 170).

[0208] In an embodiment, the peptide is bound to a protective cap group at its C-terminus and / or N-terminus. Exemplary protective groups that can be bound to the C-terminus include amides, aldehydes, esters, p-nitroaniline, and 7-amino-4-methylcoumarin. Exemplary protective groups that can be bound to the N-terminus are selected from the group consisting of acetyl, formyl, pyroglutamyl, fatty acids, urea, carbamate sulfonamides, and alkylamines.

[0209] In another embodiment, one or more amino acid residues may be added to the N-terminus or C-terminus of the peptide. In one embodiment, any of the peptides described herein may have amino acid substitutions, which are conservative substitutions, also referred to in the art as conservative mutations or conservative substitutions. Amino acid substitutions that change a given amino acid to a different amino acid provide a mutated peptide having biochemical properties similar to those of a peptide without conservative substitutions.

[0210] In an embodiment, the peptide consists of a continuous sequence of 8-50, 8-40, 8-35, 8-30, 10-40, 10-35, 10-30, 10-25, 10-20, or 12-18 amino acid residues. In an embodiment, the peptide has at least about 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 95%, 97%, 98% or 99% sequence identity with the peptide of Formula I. In another embodiment, the peptide has at least about 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 95%, 97%, 98% or 99% sequence identity with the peptide of SEQ ID NO: 1-166. In another embodiment, the peptide has at least about 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 95%, 97%, 98% or 99% sequence identity to the peptide of SEQ ID NO: 1-214.

[0211] The IL-17 inhibitor may also be a dimer of two such peptides or a bioconjugate comprising an IL-17 inhibitory peptide linked to a biomolecule.

[0212] In an embodiment, the peptide is in the form of a dimer formed by two peptides, wherein each peptide is a peptide of Formula I. In one embodiment, the two peptides in the dimer are linked by a polyethylene spacer.

[0213] In another embodiment, the peptide is in the form of a bioconjugate comprising a peptide of formula I and a biomolecule, wherein the biomolecule is bound to the N-terminus and / or C-terminus of the peptide. The bioconjugate may optionally include a linker between the peptide and the biomolecule. Exemplary biomolecules include macromolecules (e.g., carbohydrates, lipids, and proteins) or small natural products. Other exemplary biomolecules are ascorbic acid, capric acid, caproic acid, N-acetylglucosamine (NAG), N-acetylmuramic acid (NAM), NAG-NAM, hyaluronic acid, alginic acid, chitin, (GalNAc) 2, Gal-α1,3-GalNAc, and trigalacturonic acid.

[0214] The IL-17 inhibitor can be formulated into a composition for application to the eye. In an embodiment, the composition comprises saline or other ophthalmologically acceptable buffer. In other embodiments, the composition comprises one or more ophthalmologically acceptable excipients, such as alcohol (ethanol, propanol and nonanol), fatty alcohol (lauryl alcohol), fatty acid (valeric acid, caproic acid and capric acid), fatty acid ester (isopropyl myristate and isopropyl n-hexanoate), alkyl ester (ethyl acetate and butyl acetate), polyol (propylene glycol, propylene glycol and hexanetriol), sulfoxide (dimethyl sulfoxide and decyl methyl sulfoxide), amide (urea, dimethylacetamide and pyrrolidone derivatives), surfactant (sodium dodecyl sulfate, hexadecyl trimethylammonium bromide, poloxamer (polaxamer), span (span), tween, bile salts and lecithin), terpenoids (d-limonene, α-terpene alcohol, 1,8-cineole and menthone) or alkanone (n-heptane and n-nonane).

[0215] Optionally, the composition further contains a compound selected from the group consisting of: physiologically acceptable salts, poloxamer analogs and carbopol, carbopol / hydroxypropyl methylcellulose (HPMC), carbomer-methylcellulose, carboxymethylcellulose (CMC), hyaluronic acid, cyclodextrin and petroleum. Bioadhesive compositions, such as those comprising polymers, are also contemplated. Exemplary mucoadhesive polyanionic natural or semisynthetic polymers include, but are not limited to, polygalacturonic acid, hyaluronic acid, carboxymethyl amylose, carboxymethyl chitin, chondroitin sulfate, heparin sulfate and interstitial polysaccharides. Suitable hydrogels include polylactic acid, polyglycolic acid, PLGA polymers, alginate and alginate derivatives, gelatin, collagen, agarose, natural and synthetic polysaccharides, polyamino acids (such as polypeptides, especially poly(lysine)), polyesters (such as polyhydroxybutyrate and poly-ε-caprolactone), polyanhydrides; polyphosphazines, poly(vinyl alcohol), poly(alkylene oxides) (especially poly(ethylene oxide)), poly(allylamine) (PAM), poly(acrylates), modified styrene polymers (such as poly(4-aminomethylstyrene)), pluronic polyols, poloxamers, poly(uronic acid), poly(vinyl pyrrolidone) and copolymers of the above, including graft copolymers. In another embodiment, synthetic polymers and naturally occurring polymers are contemplated, such as, but not limited to, collagen, fibrin, hyaluronic acid, agarose, and laminin-rich gels.

[0216] The composition is applied to the eye. In an embodiment, the composition is directly applied by topical application to the ocular surface. In another embodiment, the composition is directly applied by topical application to the conjunctival sac. In another embodiment, the composition is applied by instillation into or onto the eye. In an embodiment, the composition is instilled into the conjunctival sac or onto the ocular surface in one or more drops.

[0217] The dosage of the IL-17 inhibitor will vary depending on the compound, its potency, and other factors. In embodiments, the composition provides a therapeutically effective amount of an IL-17 inhibitor. For example, if the compound is an IL-17A peptide inhibitor, a therapeutically effective amount is typically a total daily dose of about 0.01-1,000 μM, or about 0.05-750 μM, or about 0.05-500 μM, or a total daily dose of at least about 500 μM or 1,000 μM.

[0218] Dry Eye Questionnaire

[0219] In embodiments, the method includes identifying a subject with dry eye and / or quantifying the severity of dry eye. In these embodiments, a questionnaire can be used to identify a subject with dry eye or quantify the severity of dry eye. In another embodiment, a questionnaire is used to assess the progression of dry eye, and more specifically, whether the treatment delays, slows or inhibits the progression of mild or moderate dry eye to moderate or severe dry eye.

[0220] An exemplary questionnaire is the Visual Analog Scale (VAS)-Symptom Index. The VAS questionnaire asks questions about ocular discomfort and requires the subject to subjectively assess each ocular symptom by placing a vertical mark on a horizontal line to indicate the degree of discomfort, with 0% corresponding to no discomfort and 100% corresponding to maximum discomfort. The ocular symptoms inquired about are burning / stinging, itching, foreign body sensation, eye discomfort, dry eyes, photophobia, and pain.

[0221] Another exemplary questionnaire is the Ocular Surface Disease Index (OSDI; Allergan Inc., Irvine, CA; Walt JG et al., Drug Inf J., 1997; 31: 1436), which is a commonly used tool for assessing dry eye disease (Amparo, F. et al., Ophthalmology, 2015: 122(7): 1498-1503). The questionnaire consists of 12 questions and assesses the frequency of symptoms in the previous week. The scores range from 0 to 100, and based on the scores, the patient's symptoms can be classified as normal (0-12), mild dry eye (13-22), moderate dry eye (23-32), or severe dry eye (33-100) (Schiffman RM et al., Arch Ophthalmol., 2000; 118: 615-21; Miller KL, Arch Ophthalmol., 2010; 128: 94-101).

[0222] The Dry Eye Symptom Assessment Questionnaire (SANDE) is a questionnaire based on a visual analog scale that quantifies the severity and frequency of dry eye symptoms. The SANDE consists of two questions, and each question uses a 100 mm horizontal linear visual analog scale. The measurement range of symptom frequency ranges from "rare" to "always", and the symptom severity ranges from "very mild" to "very severe". (Gulati A. et al., Am J Ophthalmol., 2006; 142: 125-31; Schaumberg DA et al., OculSurf., 2007; 5: 50-7). Both tests are reliable and valid measures of dry eye symptoms (Schiffman R.M. et al., Arch Ophthalmol., 2000; 118: 615-21; Gulati A. et al., Am J Ophthalmol., 2006; 142: 125-31). The first question in the SANDE questionnaire explores the frequency of symptoms by asking the subject to place an "x" on the line from "rarely" to "always" to indicate how often, on average, the person experiences eye dryness and / or irritation. The second question rates the severity of symptoms by asking the subject to place an "x" on the line from "very mild" to "very severe" to indicate how severely, on average, the person experiences dryness and / or irritation.

[0223] The Dry Eye Questionnaire (DEQ) diagnoses dry eye disease and quantifies its severity. The questionnaire measures several symptoms using four variables: irritation level, frequency, morning intensity, and intensity later in the day (Chalmers RL et al., Contact Lens and Anterior Eye, 2010; 33(2):55-60). The DEQ allows differentiation of ophthalmic status: patients with dry eye disease versus patients without dry eye disease, patients with Sjögren's syndrome and keratoconjunctivitis sicca versus patients without Sjögren's syndrome and keratoconjunctivitis sicca, and controls versus patients with Sjögren's syndrome and keratoconjunctivitis sicca versus patients without Sjögren's syndrome and keratoconjunctivitis sicca (Begley CG et al., Cornea, 2002; 21(7):664-67).

[0224] The SPEED questionnaire was designed by Korb and Blackie to quickly track the progression of dry eye symptoms over time (Blackie, C. et al., Ocular Surgery News, Europe Edition 2012). The SPEED questionnaire gives a score of 0 to 28, which is the result of evaluating the frequency and severity of symptoms. The symptoms evaluated include dryness, gritty sensation or itching; pain or irritation; burning or moistening; and eye fatigue. The questionnaire measures the frequency of each symptom by asking whether the symptom never occurs (0), sometimes occurs (1), often occurs (2), or occurs continuously (3). The severity is assessed by asking whether the symptoms are no problem (0), tolerable (1), uncomfortable (2), annoying (3), or unbearable (4), where tolerable is defined as "not perfect, but not uncomfortable"; uncomfortable is "annoying, but does not interfere with my life", annoying is "annoying and interferes with my life", and unbearable is "unable to carry out my daily tasks".

[0225] Signs of dry eye can be measured in various ways, including staining (e.g., corneal fluorescein staining and lissamine green conjunctival staining), analysis of biomarkers, tear film breakup time, Schirmer test (with or without anesthesia) or conjunctival hyperemia. Staining measures for evaluating dry eye include corneal staining (e.g., using fluorescein or lissamine, etc.), and grading staining using a system (e.g., the National Eye Institute (NEI) grading system or other scales). An exemplary NEI scale is a fluorescein staining scale that uses a standardized grading system of 0 to 3 for each of the five areas of each cornea. A standard exemplary NEI system uses a 0-15 scale of fluorescein staining; another NEI system uses a 0-18 scale and lissamine green conjunctival staining. The methods described herein may additionally include using one or more of these measures to measure, monitor or determine signs of dry eye and dry eye. The methods described herein may additionally include using one or more of these measures to identify subjects with dry eye.

[0226] In one embodiment, the subject with dry eye is a subject who does not have dry eye due to Sjögren's syndrome, meibomian gland dysfunction, uveitis, an intraocular condition, or inflammation of tissues inside the eye. In other embodiments, neither the IL-17 inhibitor nor the IL-17 cytokine inhibitor is a peptide having a sequence identified herein as SEQ ID NO: 167-214.

[0227] III. Example

[0228] The following examples are illustrative in nature and are not intended to be limiting in any way.

[0229] Example 1

[0230] Treatment of dry eyes

[0231] Such a study was conducted to compare the efficacy of IL-17 cytokine inhibitor, lifitegrast ophthalmic solution and cyclosporine ophthalmic emulsion Treatment of dry eye disease. The IL-17 cytokine inhibitor is an anti-IL-17A antibody (R&D Systems, Inc. clone 50105), formulated as 1wt% in phosphate buffered saline. Female C57BL / 6 (Charles River Laboratories) aged 6 to 8 weeks were obtained. Dry eye disease was induced by placing mice in a controlled environment room with a relative humidity of <20%, an airflow of 15L / min, and a constant temperature of 21-23°C for 14 days. Corneal epithelial disease was assessed using corneal fluorescein staining (CFS) on day 0 (normal baseline), day 4 (before topical treatment instillation), day 7, day 10, and day 14, and the National Eye Institute Industry Workshop Scale (0-15 points) was used for scoring.

[0232] After induction of dry eye disease, the mice were randomly assigned to cohorts for the following treatments: (i) anti-IL-17A antibody once daily; (ii) anti-IL-17A antibody twice daily; (iii) lifitegrast twice daily; (iv) cyclosporine twice daily; or (v) saline twice daily as a control. Test compounds were applied topically to the eyes for 12 days, and disease severity was assessed daily using the National Eye Institute Industry Workshop Scale. Results Figure 1 shown.

[0233] Submandibular and cervical draining lymph nodes were obtained from the mice and single cell suspensions were prepared. 182 The CD4+CD25+Foxp3+Treg cells and total CD4+T cells of the suspension were analyzed as described in :1247(2009). Figure 2A-2B shown.

[0234] Eye tissue was also obtained from the mice and tissue sections were prepared for microscopic analysis. Figures 3A-3E As shown, healthy mice without dry eye disease ( Figure 3A ) and mice with dry eye treated twice daily with: saline as a control ( Figure 3B ), Cyclosporine ophthalmic emulsion ( Figure 3C ), lifitegrast ophthalmic solution ( Figure 3D ) or anti-IL-17A antibody ( Figure 3E ). Figure 3F is a bar graph of the percentage of goblet cells for normal (healthy) mice and mice with dry eye disease treated with saline, cyclosporine, lifitegrast, or an IL-17A inhibitor.

[0235] Example 2

[0236] Treatment of dry eyes

[0237] Co-cultures of human corneal epithelial cells and Th17 cells were prepared to mimic the in vivo interaction between Th17 cells and corneal tissue in dry eye disease. The therapeutic effect of IL-17 inhibitors in protecting human corneal epithelial cells from Th17-induced damage was tested. Immortalized human corneal epithelial cells were obtained. To obtain human Th17 cells, naive CD4+ T cells were first purified from primary human peripheral blood mononuclear cells (StemCell) by negative selection using MACS separation, and then polarized into Th17 cells in vitro using a human Th17 cell differentiation kit (RnD Systems) in X-VIVO 15 medium (Lonza). The successful differentiation of human Th17 cells was confirmed by flow cytometry before co-culture. When the human corneal epithelial cell culture was approximately 80% confluent, 1×10 5 After 24 hours of co-culture in the presence of the IL-17 inhibitor, the density and morphology of human corneal epithelial cells were observed under a bright light inverted microscope equipped with a camera. Representative images are shown in Figures 4A-4C shown.

[0238] Example 3

[0239] Treatment of dry eyes

[0240] Co-cultures of human corneal epithelial cells and Th17 cells were prepared to mimic the in vivo interaction between Th17 cells and corneal tissue in dry eye disease. The therapeutic effect of IL-17 inhibitors in protecting human corneal epithelial cells from Th17-induced damage was tested. Immortalized human corneal epithelial cells were obtained. To obtain human Th17 cells, naive CD4+ T cells were first purified from primary human peripheral blood mononuclear cells (StemCell) by negative selection using MACS separation, and then polarized into Th17 cells in vitro using a human Th17 cell differentiation kit (RnD Systems) in X-VIVO 15 medium (Lonza). The successful differentiation of human Th17 cells was confirmed by flow cytometry before co-culture. When the human corneal epithelial cell culture was approximately 80% confluent, 1×10 5Th17 cells. The IL-17 inhibitor is a 15 amino acid peptide identified as SEQ ID NO: 1 (VHVTIPADLWDWINK) that inhibits the binding of IL-17A to the IL-17A receptor. The IL-17 inhibitor peptide is formulated using phosphate buffered saline at an appropriate concentration to provide a composition with an IL-17 inhibitor at a concentration of 0.1 μM, 1 μM, 10 μM, and 100 μM.

[0241] After 24 h of co-culture in the presence of each treatment dose of IL-17 inhibitor, the density and morphology of human corneal epithelial cells were observed under a bright light inverted microscope equipped with a camera. Representative images are shown in Figures 5A-5F shown.

[0242] Example 4

[0243] Slowing the progression of dry eye disease

[0244] Female C57BL / 6 mice aged 6 to 8 weeks were used in this study, and dry eye was induced as described in Example 1. Four days after induction of dry eye, the mice were randomly assigned to 4 treatment cohorts (n=10) for treatment with saline or with an IL-17 inhibitor. Two IL-17 inhibitors were tested: an anti-IL-17A antibody (R&D Systems, Inc. clone 50105) formulated at 1 wt% in phosphate-buffered saline and a 15 amino acid peptide identified as SEQ ID NO: 1 (VHVTIPADLWDWINK) that inhibits the binding of IL-17A to the IL-17A receptor formulated in phosphate-buffered saline at a dose of 0.05 wt% or 0.1 wt%. The treatment composition and saline control were topically administered by instilling 3 μL into the eye twice a day from days 4-12. Corneal epithelial disease was assessed using corneal fluorescein staining (CFS) and scored using the National Eye Institute Industry Workshop Scale (0-15 points) on days 0 (normal baseline), 4 (before topical treatment instillations), 7, 10, and 12. Fig. 6A shown.

[0245] At the end of treatment, ocular draining lymph nodes were collected and analyzed for the frequency of IL-17+CD4+Th17 cells by flow cytometry. The percentage of Th17 cells in the draining lymph nodes of the treatment cohort was as follows: Figure 6B shown.

[0246] Example 5

[0247] Slowing the progression of dry eye disease

[0248] Female C57BL / 6 mice aged 6 to 8 weeks were used in this study, and dry eye was induced as described in Example 1. Four days after induction of dry eye, the mice were randomly assigned to treatment cohorts (n=6) for treatment with saline or with an IL-17 inhibitor. The IL-17 inhibitor was a 15-amino acid peptide identified as SEQ ID NO: 1 (VHVTIPADLWDWINK) that inhibits the binding of IL-17A to the IL-17A receptor, formulated in phosphate-buffered saline at concentrations of 50 μM, 250 μM, and 500 μM. A formulation containing 250 μM IL-17A inhibitor was administered once a day to one treatment cohort and twice a day to a second treatment cohort. A formulation containing 50 μM IL-17A inhibitor was administered twice a day to one treatment cohort, and a formulation containing 500 μM was administered once a day to one cohort. The treatment composition and saline control were topically administered by instilling 3 μL into both eyes of the subject. Table 5-1 summarizes the treatment cohorts and study designs.

[0249] Table 5-1

[0250]

[0251] Corneal epithelial disease was assessed using corneal fluorescein staining (CFS) and scored using the National Eye Institute Industry Workshop Scale (0-15 points) on days 0 (normal baseline), 3 (before topical treatment instillation), 7, and 10. The cohort treated with the formulation containing 250 μM IL-17A inhibitor was treated for 21 days, with CFS and scoring performed on days 3, 7, 10, 14, 17, and 21. Results Figure 7 and Figure 8 shown.

[0252] Example 6

[0253] Restoration of tear film homeostasis

[0254] Human subjects with moderate to severe dry eye disease were enrolled in a study treated with an IL-17 inhibitor. The IL-17 inhibitor was applied directly to the subject's eye daily for 1 month. Ocular surface assessments were performed prior to the first dose and then weekly.

[0255] Representative schematic diagrams of 5 corneal and 6 conjunctival areas per eye evaluated for ocular surface staining with fluorescein and lissamine green using the National Eye Institute Industry Workshop scale and recorded on a case report form. The case report form includes a graphic representation and a descriptive rating scale (rating 0 to 15, or 0 to 18).

[0256] 1. Evaluate corneal staining using 1.0 mg sodium fluorescein test strips.

[0257] 2. After wetting the end of the test strip with a single drop of buffered saline, quickly flick the excess with your fingers into a trash can.

[0258] 3. The lower eyelid is then pulled down and the flat end of the tip is gently applied to the lower tarsal conjunctiva, aiming to instill a very small amount of dye and not induce reflex tearing.

[0259] 4. Instruct the patient to blink naturally several times without forcibly closing the eyelids to distribute the fluorescein.

[0260] 5. After allowing the fluorescein to remain on the eye for at least one minute, five corneal areas will be graded using a yellow (Wratten #12) barrier filter in combination with a cobalt (blue) filter to maximize the fluorescent field of view. The upper eyelid is slightly lifted so that the entire corneal surface can be graded. To enhance contrast, a yellow barrier filter is placed in the path of the returning light (not in the path of the incident light).

[0261] Tear Film Breakup Time (TFBUT) will be assessed using a slit lamp biomicroscope according to the following steps:

[0262] 1. The slit lamp will be set to approximately 10x magnification.

[0263] 2. With adequate fluorescein placed (preferably using DET test strips), the subject will be asked to look straight ahead and not blink until further notice. The test should be performed in a room with no direct air flow on the patient's face.

[0264] 3. A stopwatch will be used to record the time from the last complete blink to the first appearance of growing micelles indicating tear film breakup. If the patient blinks prematurely prior to the breakup of the mire, the examiner should continue to attempt to obtain a reading.

[0265] 4. Once the TFBUT is observed, instruct the patient to blink freely. The test should then be repeated a second time on the same eye.

[0266] 5. If the difference between the first and second readings differs by more than two seconds, a third measurement should be made and recorded.

[0267] 6. Then do this procedure in the other eye.

[0268] 7. It is recommended to conduct TFBUT in a room with a temperature of about 18°C ​​and a humidity of about 50%.

[0269] Ocular surface staining assessment will be completed using lissamine green conjunctival stain.

[0270] 1. Lissamine Green Eye Test Strips should be moistened with buffered saline and applied to the lower tarsal conjunctiva. Care should be taken to instill sufficient dye.

[0271] 2. After allowing lissamine green to remain on the eye for one minute, the six nasal conjunctival areas and the temporal conjunctival area will be graded.

[0272] 3. To grade the temporal region, the subject should be instructed to look toward the nose; to grade the nasal region, the subject should be instructed to look toward the temporal region.

[0273] 4. The procedure should then be completed in the other eye.

[0274] The present study demonstrated that long-term treatment with the described IL-17 inhibitor improved TFBUT, indicating restoration of tear film homeostasis.

[0275] Example 7

[0276] Methods for treating dry eye disease

[0277] Human subjects with moderate to severe dry eye disease were enrolled in a study treated with an IL-17 inhibitor. The IL-17 inhibitor was a spirocyclic indane compound that was applied directly to the subject's eyes daily for 1 month. Ocular surface and tear film assessments were performed as described in Example 6 prior to the first dose and then weekly.

[0278] Although many exemplary aspects and embodiments have been discussed above, those skilled in the art will recognize certain modifications, permutations, additions and sub-combinations thereof. Therefore, the appended claims and claims introduced below should be interpreted to include all such modifications, permutations, additions and sub-combinations within their true spirit and scope.

Claims

1. A method of treating dry eye in a subject diagnosed with dry eye, the method comprising: A composition comprising a therapeutically effective amount of a compound that inhibits the binding of an inflammatory interleukin 17 (IL-17) cytokine to an IL-17 receptor is administered directly to the eye of a subject in need thereof.

2. A method for slowing, inhibiting or delaying the progression of dry eye disease in a subject, the method comprising: A composition comprising a therapeutically effective amount of a compound that inhibits the binding of an inflammatory interleukin 17 (IL-17) cytokine to an IL-17 receptor is administered directly to the eye of a subject in need thereof.

3. A method of reversing the progression of dry eye disease in a subject, the method comprising: A composition comprising a therapeutically effective amount of a compound that inhibits the binding of an inflammatory interleukin 17 (IL-17) cytokine to an IL-17 receptor is administered directly to the eye of a subject in need thereof.

4. A method for restoring tear film homeostasis on the surface of the eye, the method comprising: A composition comprising a therapeutically effective amount of a compound that inhibits the binding of an inflammatory interleukin 17 (IL-17) cytokine to an IL-17 receptor is administered directly to the eye of a subject in need thereof.

5. The method of any one of claims 1-4, further comprising identifying a subject suffering from mild or moderate dry eye disease.

6. The method of any one of claims 1-4, further comprising identifying a subject suffering from mild, moderate or severe dry eye disease.

7. A method according to claim 5 or claim 6, wherein the identification comprises the use of a questionnaire.

8. The method of claim 7, wherein the questionnaire is selected from the group consisting of a visual analogue scale (VAS), an ocular surface disease index (OSDI) questionnaire, a symptom assessment questionnaire for dry eye (SANDE) questionnaire, a dry eye questionnaire (DEQ), and a standard patient evaluation questionnaire for dry eye (SPEED).

9. The method of any one of claims 2, further comprising identifying subjects with mild or moderate dry eye disease using the Standard Patient Evaluation of Dry Eye Questionnaire (SPEED) and monitoring the progression of dry eye disease.

10. The method of claim 9, wherein the identification comprises the use of a questionnaire.

11. The method according to any one of the preceding claims, wherein the administering comprises administering a composition comprising a compound selected from a biomolecule or an organic synthetic compound.

12. The method according to claim 11, wherein the organic synthetic compound is a spirocyclic indane compound or a spirocyclic oxoindole compound.

13. The method of claim 11, wherein the biological compound is an antibody having binding affinity for IL-17 cytokine or an antibody having binding affinity for IL-17 receptor.

14. The method of claim 13, wherein the antibody is a monoclonal antibody, a polyclonal antibody, a single-chain antibody, a humanized antibody, a recombinant antibody, a chimeric antibody or an antibody fragment.

15. The method of any one of claims 13-14, wherein the antibody is selected from the group consisting of afasevikumab, bimekizumab, brodalumab, ixekizumab, izokibep netakimab, perakizumab, secukinumab, sonelokimab, tibulizumab, vunakizumab, ABY-035, CJM-112, CNTO-6785, DC-806 / S-011806, FPP -003, GR-1501, HB-0017, IMU-035, LZM-012, QX-002N, BH-1657, HB-0043, HT-0017, ILCT-100 1. IQ-001, LEO153339 / LP0200, LP-0200, MT-6194, MYMD-1, ND-016, SCT-650A, SM-17, YBL-00 4. ABM-60, ETI-1023, LQ-025, LQ-026, ABBV-257, AFB-035, ANB-004, BCD-121, COVA-322, CYT -017-IL17Qb, DLX-2882, DLC-2907, DLX-2909, DLX-3003, E-34935, E-35018, E-35762, E-360 41. EBI-006, HEISCO-III-002, IL-17-RC, MEDI-571, MOR-106, MP-0230, PRS-190, SCH-90011 7. Y-320, ABT-122, BITS-7201A, CDP-435, EBI-028, IL-17E, JNJ-6118104, KHK-4827 and RG7624.

16. The method of claim 11, wherein the biological compound is a protein or peptide that specifically inhibits the binding of IL-17A to the interleukin 17A receptor.

17. The method of claim 16, wherein the peptide consists of a contiguous sequence of 12-18 amino acid residues having at least about 70% sequence identity with SEQ ID NO:

1.

18. The method of claim 16, wherein the peptide comprises an amino acid sequence of Formula I: X1-X2-X3-X4X5-X6-X7-X8-X9-X 10 -X1I-X 12 -X 13 -X 14 -X 15 Formula (I), in X1 is I, V or L; X2 is H, M, R, K or E; X3 is V, F or I; X4 is T, Q, S, Y or N; X5 is I, F, or V; X6 is P or G; X7 is A, Q or L; X8 is D, E or Q; X9 is L, W, F, V, or I; X 10 is W, Y or F; X 11 is D, E or N; X 12 is W or F; X 13 is I, V, F, or L; X 14 is N, R, Q or E; and X 15 It is K, R, H or E.

19. The method according to claim 18, wherein (a) X1 is I or V, X2 is H, M or R, X3 is V or F; X4 is T or Q; X5 is I, F, or V; X6 is P or G; X7 is A or Q; X8 is D or E; X9 is L; X 10 is W or Y; X is D or E; X 12 It is W; X 13 is I or V; X 14 is N, R or E; and X 15 is K, R or E; or (b) X1 is I or V; X2 is H or M, X3 is V; X4 is T; X5 is I; X6 is P; X7 is A; X8 is D; X9 is L, W, F, V, or I; X 10 is W or Y; X 11 is D or E; X 12 It is W; X 13 is I or V; X 14 is N, R or E; and X 15 It is K, R or E.

20. The method according to any one of claims 17-19, wherein the administering comprises administering a peptide having a protective cap group bound to its C-terminus and / or N-terminus, wherein the protective cap group bound to the C-terminus is selected from the group consisting of amides, aldehydes, esters, p-nitroaniline, 7-amino-4-methylcoumarin, and the protective cap group bound to the N-terminus is selected from the group consisting of acetyl, formyl, pyroglutamyl, fatty acids, urea, carbamate sulfonamides, and alkylamines.

21. The method of any one of claims 17-20, wherein the administering comprises administering the peptide in the form of a dimer formed by two peptides, each of which is a peptide of Formula I.

22. The method of claim 21, wherein the two peptides in the dimer are linked by a polyethylene spacer.

23. The method of any one of claims 17-22, wherein the administering comprises administering the peptide in the form of a bioconjugate comprising the peptide of Formula I and a biomolecule, wherein the biomolecule is bound to the N-terminus and / or C-terminus of the peptide.

24. The method of claim 23, wherein the biomolecule is selected from the group consisting of decanoic acid, hexanoic acid, ascorbic acid, NAG-NAM, NAG, NAM, hyaluronic acid, alginic acid, chitin, (GalNAc)2, Gal-α1,3-GalNAc and trigalacturonic acid.

25. The method of any one of claims 17-24, wherein the administering comprises administering the peptide in a pharmaceutically acceptable composition comprising at least one pharmaceutically acceptable excipient.

26. The method of any one of the preceding claims, wherein the administering comprises instillation onto the ocular surface.

27. The method of any one of the preceding claims, wherein the administering comprises instillation into the conjunctival sac.

28. The method of any one of the preceding claims, wherein the administering comprises administering once daily.

29. The method of any one of claims 17-28, wherein the therapeutically effective dose of the peptide is at least about 500 μM.

30. The method of claim 29, wherein the dose is administered once daily.

31. The method of any one of the preceding claims, wherein the administering continues for a period of at least about 3 weeks, wherein dry eye signs and symptoms resolve during a disease-free period of the third week of the administering.

32. A method of restoring goblet cell function and / or density in an eye having signs or symptoms of dry eye disease, or a method of preventing goblet cell loss in an eye of a person suffering from dry eye disease, the method comprising: A therapeutically effective amount of a compound that inhibits the binding of IL-17A to the interleukin 17A receptor is topically administered to the eye of a subject in need thereof.

33. A method for improving the balance of lacrimal gland, meibomian gland and conjunctival goblet cell function to restore tear film homeostasis in a subject, the method comprising: A therapeutically effective amount of a compound that inhibits the binding of IL-17A to the interleukin 17A receptor is topically administered to the eye of a subject in need thereof.

34. A method of increasing basal tear production in an elderly subject, the method comprising: A therapeutically effective amount of a compound that inhibits the binding of IL-17A to the interleukin 17A receptor is topically administered to the eye of a subject in need thereof.

35. The method of claim 34, wherein the elderly subject has hormonal changes or is over 50 years of age.

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