WO2018035615A1

WO2018035615A1 - Compositions and methods for the treatment of inflammatory bowel diseases

Info

Publication number: WO2018035615A1
Application number: PCT/CA2017/051001
Authority: WO
Inventors: Roger Leger; Jerome ROSSERT; Marie-Elaine Caruso
Original assignee: Thrasos Therapeutics Inc.
Priority date: 2016-08-26
Filing date: 2017-08-25
Publication date: 2018-03-01

Abstract

Provided are novel peptides of Formula SEQ ID No. 1: J¹CysX¹X²X³X⁴X⁵X⁶,ProX⁷ThrCysJ²J³(J⁴)₅(J⁵)₁; (SEQ ID No. 1 ) pharmaceutically acceptable salts thereof, and pharmaceutical compositions thereof, which are which are effective inhibitors of light chains to uromodulin.

Description

COMPOSITIONS AND METHODS FOR THE TREATMENT OF

INFLAMMATORY BOWEL DISEASES

RELATED APPLICATIONS

[0001 ] This application claims priority to U.S. Provisional Application No.

62/380,075, filed August 26, 2016, the entire contents of which are incorporated herein by reference.

BACKGROUND

[0002] Inflammatory bowel diseases (IBDs) arc a group of inflammatory disorders of the colon and small intestine. Autoimmunity Reviews. 2010;9(5):A372-A378. Of the IBDs, Crohn's disease (CD) and ulcerative colitis (UC) remain the most prevalent. Although the pathogenesis of CD and UC is not fully understood, these disorders continue to affect nearly 1 to 1.3 million people in the United States alone.

[0003] Granule membrane glycoprotein 2 (GP2) has been identified as die major autoantigen of CD-specific pancreatic autoantibodies (PAB). See e.g., Gastroenterol Res Pract. 2013; 2013: 683824. GP2 has also been found to be overexpressed in patient populations having ulcerative colitis. See e.g., Gastroenterology Research and Practice Volume 2013 (2013), Article ID 683824. It was not until recently, however, that modulation of GP2 was identified as a target for not only differentiating and having therapeutic impacts on CD and UC, but also for stimulating adaptive immune responses. See e.g., J Immunol 2012; 189:2774-2783 and Nature Letters, Vol. 462, 12, November 2009.

[0004] Targeting GP2, therefore, represent an attractive approach to combat die unmet medical need for improved agents against inflammatory bowel diseases, particularly Crohn's disease and ulcerative colitis.

SUMMARY OF THE INVENTION

[0005] Provided herein arc peptides and compositions thereof that recognize GP2. Such peptides include those of Formula SEQ ID No. 1:

or a pharmaceutically acceptable salt thereof, wherein each of X¹, X², X³, X⁴, X^s, X⁶, X⁷, J¹, J² J³, J⁴, J⁵, s, and t are as defined and described herein.

[0006] Also provided are methods of treating inflammatory bowel diseases such as Crohn's disease and ulcerative colitis, comprising administering one or more of the peptides or compositions described herein.

DETAILED DESCRIPTION OF CERTAIN EMBODIMENTS

/. General Description of Peptides

[0007] In one aspect, provided herein are peptides of Formula SEQ ID No. 1 :

or a pharmaceutically acceptable salt thereof, wherein

J¹ is Aha or Arg;

X¹ to X⁷ are each independently a natural or non-natural amino acid;

s, and t arc each independently 0 or 1;

J² and J³ are each independently Lys or Arg;

J⁴, if present is Lys or Arg; and

R⁵, if present is Lys or Arg.

2. Peptides and Definitions

[0008] "Amino acid" refers to an organic compound containing an amine (-NH₂) and a carboxylic acid (-COOH) functional group, usually along with a side-chain specific to each amino acid. Amino acids can be classified according to the core structural functional groups' locations as alpha- (a-), beta- (β-), gamma- (γ-) or delta- (δ-) amino acids; other categories relate to polarity, pH level, and side-chain group type (aliphatic, acyclic, aromatic, containing hydroxyl or sulfur, etc.). In one embodiment, amino acid includes

[0009] "Natural amino acid" is used interchangeably with proteinogenic amino acid and refers to the 20 standard amino acids encoded by the universal genetic code along with selenocysteine and pyrrolysine. The 20 standard amino acids encoded by the universal genetic code include glycine, alanine, valine, leucine, isoleucine, proline, phenylalanine, tyrosine, tryptophan, serine, threonine, cysteine, methionine, asparagine, glutamine, aspartate, glutamate, lysine, arginine, and histidine. Unless otherwise specified, when a natural amino acid is defined without indicating the stereochemistry it will be understood that the defined amino acid is present as the L-enantiomer. Thus, when used as part of a formula, the letter A or Ala means L-alanine.

[0010] The full name of a natural amino acid and its corresponding three-letter or one- letter code are used interchangeably. For example, glycine is interchangeable with three letter abbreviation Gly or the one letter abbreviation G; alanine is interchangeable with three letter abbreviation Ala or the one letter abbreviation A, valine is interchangeable with three letter abbreviation Val or the one letter abbreviation V, leucine is

interchangeable with three letter abbreviation Leu or the one letter abbreviation L, isoleucine is interchangeable with three letter abbreviation He or the one letter

abbreviation I, proline is interchangeable with three letter abbreviation Pro or the one letter abbreviation P, phenylalanine is interchangeable with three letter abbreviation Phe or the one letter abbreviation F, tyrosine is interchangeable with three letter abbreviation Tyr or the one letter abbreviation Y, tryptophan is interchangeable with three letter abbreviation Trp or the one letter abbreviation W, serine is interchangeable with three letter abbreviation Ser or the one letter abbreviation S, threonine is interchangeable with three letter abbreviation Thr or the one letter abbreviation T, cysteine is interchangeable with three letter abbreviation Cys or the one letter abbreviation C, methionine is interchangeable with three letter abbreviation Met or the one letter abbreviation M, asparagine is interchangeable with three letter abbreviation Asn or the one letter abbreviation N, glutamine is interchangeable with three letter abbreviation Gin or the one letter abbreviation Q, aspartate is interchangeable with three letter abbreviation Asp or the one letter abbreviation D, glutamate is interchangeable with three letter abbreviation Glu or the one letter abbreviation E, lysine is interchangeable with three letter abbreviation Lys or the one letter abbreviation K, arginine is interchangeable with three letter abbreviation Arg or the one letter abbreviation R, histidine is interchangeable with three letter abbreviation His or the one letter abbreviation H, selenocysteine is interchangeable with three letter abbreviation Sec or the one letter abbreviation U, and pyrrolysine is interchangeable with three letter abbreviation Pyl or the one letter abbreviation O. In one embodiment, the term "natural amino acid" or proteinogenic amino acid refers only to the 20 standard amino acids encoded by the universal genetic code, i.e., G, A, V, L, I, P, F, Y, W, S, T, C, M, N, Q, D, E, K, R, and H.

[0011] "Non-natural amino acid" refers to a non-proteinogenic amino acid that is not found in proteins (e.g., carnitine, ga/n/na-aminobutyric acid, and D-forms of natural amino acids except glycine) or not produced directly and in isolation by standard cellular machinery (e.g., hydroxyprolinc and selenomethionine). Other examples include, but are not limited to, β-amino acids (β3 and β2), homo-amino acids, alanine derivatives, alicyclic amino acids, arginine derivatives, asparagine derivatives, aspartic acid derivatives, cysteine derivatives, 2,4-diaminobutyric acid, glycine derivatives, isoleucine derivatives, leucine derivatives, lysine derivatives (such as 6-aminohexanoic acid abbreviated herein as Aha), methionine derivatives, norleucine (nL) and norleucinc derivatives, phenylalanine derivatives, phenylglycine derivatives, proline and pyruvic acid derivatives, pyroglutamine derivatives, serine derivatives, threonine derivatives, tryptophan derivatives, norvaline derivatives, 2,3-diaminopropionic acid, ornithine derivatives, valine derivatives, linear core amino acids, and N-methyl amino acids. In one embodiment, the term "non-natural amino acid" refers only to D-forms of the 20 standard amino acids encoded by the universal genetic code. These forms include D-Ala, D-Val, D-Leu, D-Ile, D-Pro, D-Phe, D-Tyr, D-Trp, D-Ser, D-Thr, D-Cys, D-Met, D-Asn, D-Gln, D-Asp, D-Glu, D-Lys, D- Arg, and D-His.

[0012] A "D-amino acid" or D-form of an amino acid means that the indicated amino acid is present as the D-enantiomer. Shorthand notation for the D-enantiomer of an amino acid can be represented by an asterisk (*). For example, I* or Pie, wherein * represents a D-amino acid, refers to the D-enantiomer of He (isoleucine).

[0013] The peptides described herein may be present in the form of pharmaceutically acceptable salts. Pharmaceutically acceptable salts are art-recognized and include e.g., relatively non-toxic inorganic and organic acid addition salts, or inorganic or organic base addition salts that are suitable for human consumption. Examples of such salts include, but are not limited to, sodium, potassium, calcium, magnesium, acetate, benzoate, bicarbonate, carbonate, citrate, dihydrochloride, gluconate, glutamate, hydrochloride, and tartrate.

[0014] The term "pharmaceutically acceptable carrier" refers to a non-toxic carrier, adjuvant, or vehicle that does not adversely affect the pharmacological activity of the peptide with which it is formulated, and which is also safe for human use.

Pharmaceutically acceptable carriers that may be used include, but are not limited to, ion exchangers, alumina, aluminum stearate, magnesium stearate, lecithin, serum proteins, such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, dicalcium phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, polyvinylpyrrolidone-vinyl acetate, cellulose-based substances (e.g., microcrystalline cellulose, hydroxypropyl

methylcellulose, hydroxypropyl methylcellulose acetate succinate, hydroxypropyl methylcellulose Phthalate), starch, lactose monohydrate, mannitol, trehalose sodium lauryl sulfate, and crosscarmellose sodium, polyethylene glycol, sodium

carboxymethylcellulose, polyacrylates, polymethacrylate, waxes, polyethylene- polyoxypropylene-block polymers, polyethylene glycol and wool fat.

[001 S] The terms "subject" and "patient" may be used interchangeably, and mean a mammal in need of treatment, e.g., companion animals (e.g., dogs, cats, and the like), farm animals (e.g., cows, pigs, horses, sheep, goats and the like) and laboratory animals (e.g., rats, mice, guinea pigs and the like). Typically, the subject is a human in need of treatment.

[0016] The terms "treatment," "treat," and "treating" refer to reversing, alleviating, or inhibiting the progress of a disease or disorder, or one or more symptoms thereof, as described herein. In some embodiments, treatment may be administered after one or more symptoms have developed, i.e., therapeutic treatment. In other embodiments, treatment may be administered in the absence of symptoms. For example, treatment may be administered to a susceptible individual prior to the onset of symptoms (e.g., in light of a history of symptoms and/or in light of genetic or other susceptibility factors), i.e., prophylactic treatment. Treatment may also be continued after symptoms have resolved, for example to prevent or delay their recurrence.

[0017] An "effective amount" or "therapeutically effective amount" is a quantity sufficient to achieve a desired therapeutic and/or prophylactic effect, for example, an amount which results in the prevention of or a decrease in the symptoms associated with a condition that is being treated, e.g., the conditions described herein.

3. Description of Exemplary Peptides

[0018] In a first embodiment, provided herein are peptides of the Formula SEQ ID No. l :

or a pharmaceutically acceptable salt thereof, wherein the variables are as described above.

[0019] In a second embodiment, X⁶ in the peptide of Formula SEQ Π> No. 1 is not Glu, Thr, Asn, or Asp; and X⁷ is not Glu or Asp, wherein the remaining variables in SEQ ID No. 1 are as described above.

[0020] In a third embodiment, X¹ in the peptide of Formula SEQ ID No. 1 is selected from Arg, Lys, His, Pro, Cys, Thr, Ser, Gin, Glu, Leu, Ile, Met, Ala, Val, Gly, n-Leu, Met, Asp, and He, wherein the remaining variables in SEQ ID No. 1 are as described above for SEQ ID No. 1 or the second embodiment. Alternatively, X¹ in the peptide of Formula SEQ ID No. 1 is selected from Arg, Gin, Glu, Leu, n-Leu, Met, Asp, and He, wherein the remaining variables in SEQ ID No. 1 are as described above for SEQ ID No. 1 or the second embodiment. In another alternative, X¹ in the peptide of Formula SEQ ID No. 1 is selected from Gin, Leu, and n-Leu, wherein the remaining variables in SEQ ID No. 1 are as described above for SEQ ID No. 1 or the second embodiment.

[0021 ] In a forth embodiment, X² in the peptide of Formula SEQ ID No. 1 is selected from a natural amino acid, wherein the remaining variables in SEQ ID No. 1 are as described above for SEQ ID No. 1 or the second or third embodiment. Alternatively, X² in the peptide of Formula SEQ ID No. 1 is selected from Ser, Thr, Cys, Met, Asn, Lys, Gin, His, Arg, Glu, and Asp, wherein the remaining variables in SEQ ID No. 1 are as described above for SEQ ID No. 1 or the second or third embodiment In another alternative, X² in the peptide of Formula SEQ ID No. 1 is selected from Ser, Gin, His, Arg, Glu, and Asp, wherein the remaining variables in SEQ ID No. 1 are as described above for SEQ ID No. 1 or the second or third embodiment. In yet another alternative, X² in the peptide of Formula SEQ ID No. lis selected from Gin and His, wherein the remaining variables in SEQ Π> No. 1 are as described above for SEQ ID No. 1 or the second or third

embodiment.

[0022] In a fifth embodiment, X³ in the peptide of Formula SEQ ID No. 1 is selected from a natural amino acid, wherein the remaining variables in SEQ ID No. 1 are as described above for SEQ ID No. 1 or the second, third, or forth embodiment.

Alternatively, X³ in the peptide of Formula SEQ ID No. 1 is selected from Tyr, Trp, Phe, Ser, Thr, Cys, Met, Asn, Ala, Lys, His, Glu, Arg, and Asp, wherein the remaining variables in SEQ ID No. 1 are as described above for SEQ ID No. 1 or the second, third, or forth embodiment. In another alternative, X³ in the peptide of Formula SEQ ID No. 1 is selected from Tyr, Ser, Ala, Glu, Arg, and Asp, wherein the remaining variables in SEQ ID No. 1 are as described above for SEQ ID No. 1 or the second, third, or forth embodiment. In yet another alternative, X³ is selected from Tyr, Ser, Arg, and Asp, wherein the remaining variables in SEQ ID No. 1 are as described above for SEQ ID No. 1 or the second, third, or forth embodiment.

[0023] In a sixth embodiment, X⁴ in the peptide of Formula SEQ ID No. 1 is selected from a natural amino acid, wherein the remaining variables in SEQ ID No. 1 are as described above for SEQ ID No. 1 or the second, third, fourth, or fifth embodiment. Alternatively, X⁴ in the peptide of Formula SEQ ID No. I is selected from Gly, Ala, Val, Leu, He, Pro, Phe, Tyr, Trp, Asp, Glu, Lys, Arg, and His, wherein the remaining variables in SEQ ID No. 1 are as described above for SEQ ID No. 1 or the second, third, fourth, or fifth embodiment. In another alternative, X⁴ in the peptide of Formula SEQ ID No. 1 is selected from Leu, Asp, Gly, Tyr, Glu, and Arg, wherein the remaining variables in SEQ ID No. 1 are as described above for SEQ ID No. 1 or the second, third, fourth, or fifth embodiment. In yet another alternative X⁴ in the peptide of Formula SEQ ID No. 1 is selected from Leu, Asp, Tyr, and Arg, wherein the remaining variables in SEQ ID No. 1 are as described above for SEQ ID No. 1 or the second, third, fourth, or fifth embodiment.

[0024] In a seventh embodiment, X^s in the peptide of Formula SEQ ID No. 1 is selected from a natural amino acid, wherein the remaining variables in SEQ ID No. 1 are as described above for SEQ ID No. 1 or the second, third, fourth, fifth, or sixth embodiment. Alternatively, X^s in the peptide of Formula SEQ ID No. 1 is selected from Ser, Thr, Cys, Met, Asn, Gin, Asp, Glu, Lys, Arg, and His, wherein the remaining variables in SEQ ID No. 1 are as described above for SEQ ID No. 1 or the second, third, fourth, fifth, or sixth embodiment. In another alternative, X^s in the peptide of Formula SEQ ID No. 1 is selected from Tyr, Ser, Arg, Glu, and Asp, wherein the remaining variables in SEQ ID No. 1 are as described above for SEQ ID No. 1 or the second, third, fourth, fifth, or sixth embodiment.

[0025] In an eighth embodiment, X⁶ in die peptide of Formula SEQ ID No. 1 is selected from Gly, Ala, Val, Leu, He, Pro, Phe, Tyr, Trp, Ser, Cys, Met, Gin, Lys, Arg, and His, wherein the remaining variables in SEQ ID No. 1 are as described above for SEQ ID No. 1 or the second, third, fourth, fifth, sixth, or seventh embodiment.

Alternatively, X⁶ in the peptide of Formula SEQ ID No. 1 is selected from He, Leu, Pro, Val, Ala, Gly, His, Lys, and Arg, wherein the remaining variables in SEQ ID No. 1 are as described above for SEQ ID No. 1 or the second, third, fourth, fifth, sixth, or seventh embodiment. In another alternative, X⁶ in the peptide of Formula SEQ ID No. 1 is selected from He, Leu, and Arg, wherein the remaining variables in SEQ ID No. 1 are as described above for SEQ Π) No. 1 or the second, third, fourth, fifth, sixth, or seventh embodiment. In yet another alternative, X⁶ in the peptide of Formula SEQ ID No. 1 is selected from He and Leu, wherein the remaining variables in SEQ ID No. 1 are as described above for SEQ ID No. 1 or the second, third, fourth, fifth, sixth, or seventh embodiment.

[0026] In a ninth embodiment, X⁷ in the peptide of Formula SEQ ID No. 1 is selected from Gly, Ala, Val, Leu, He, Pro, Phe, Tyr, Tip, Ser, Thr, Cys, Met, Asn, Gin, Lys, Arg, and His, wherein the remaining variables in SEQ ID No. 1 are as described above for SEQ DD No. 1 or the second, third, fourth, fifth, sixth, seventh, or eighth embodiment. Alternatively, X⁷ in the peptide of Formula SEQ ID No. 1 is selected from Tyr, Phe, His, Lys, Arg, and Trp, wherein the remaining variables in SEQ ID No. 1 are as described above for SEQ ID No. 1 or the second, third, fourth, fifth, sixth, seventh, or eighth embodiment. In another alternative, X⁷ in the peptide of Formula SEQ ID No. 1 is selected from Tyr, Arg, and Trp, wherein the remaining variables in SEQ ID No. 1 are as described above for SEQ DD No. 1 or the second, third, fourth, fifth, sixth, seventh, or eighth embodiment. In yet another alternative, X⁷ in the peptide of Formula SEQ ID No. 1 is Tyr, wherein the remaining variables in SEQ ID No. 1 are as described above for SEQ ID No. 1 or the second, third, fourth, fifth, sixth, seventh, or eighth embodiment

[0027] In a tenth embodiment, s and t are each 0 in the peptide of Formula SEQ ID No. 1, wherein the remaining variables in SEQ ID No. 1 are as described above for SEQ Π) No. 1 or the second, third, fourth, fifth, sixth, seventh, eighth, or ninth embodiment.

[0028] In an eleventh embodiment, s and t are each 1 in the peptide of Formula SEQ Π) No. 1, wherein the remaining variables in SEQ ID No. 1 are as described above for SEQ ID No. 1 or the second, third, fourth, fifth, sixth, seventh, eighth, ninth, or tenth embodiment.

[0029] In a thirteenth embodiment, J¹ in the peptide of Formula SEQ ID No. 1 is Aha, wherein the remaining variables in SEQ ID No. 1 are as described above for SEQ ID No.

1 or the second, third, fourth, fifth, sixth, seventh, eighth, ninth, tenth, or eleventh embodiment.

[0030] Specific examples of the disclosed peptides are provided in Table 1 and Table

2 below as well as in the EXEMPLIFICATION sections. Pharmaceutically acceptable salts as well as the neutral forms of these peptides are included in the present disclosure.

Table 1

Table 2

4. Uses, Formulation, and Administration

[0031 ] In certain embodiments, the present disclosure provides a method of treating a subject (e.g., a human) suffering from IBS (e.g., UC or CD), comprising administering to the subject a therapeutically effective amount of a peptide of Formula SEQ ID No. 1, or a pharmaceutically acceptable salt thereof. In certain embodiments, the amount of peptide of SEQ ID No. 1 is such that it is an effective modulator of GP2. In certain embodiments, a provided composition is formulated for administration to a subject in need of such composition.

[0032] Pharmaceutically acceptable compositions are included and can be

administered to humans and other animals by other methods such as e.g., orally, rectally, parenterally, intracisternally, intraperitoneally, topically (as by powders, creams, ointments, or drops), bucally, as an oral or nasal spray, or the like. In certain

embodiments, the disclosed peptides may be administered parenterally (e.g., intravenous).

[0033] In certain embodiments, the present disclosure also provides a method of treating a subject (e.g., a human) suffering from IBS (e.g., UC or CD), comprising administering to the subject a therapeutically effective amount of composition comprising a peptide of Formula SEQ ID No. 1, or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable carrier, adjuvant, or vehicle. In certain embodiments, the amount of peptide of SEQ ID No. 1 in a provided composition is such that it is an effective modulator of GP2.

[0034] It will be understood that a specific dosage for any particular subject will depend upon a variety of factors, including age, body weight, general health, sex, diet, time of administration, rate of excretion, drug combination, the judgment of the treating physician, and the severity of the particular disease being treated. The amount of a provided peptide in the composition will also depend upon the particular peptide in the composition.

[003 S] The amount of provided peptides that may be combined with carrier materials to produce a composition in a single dosage form will vary depending upon the patient to be treated and the particular mode of administration.

EXEMPLIFICATION

[0036] Standard solid phase techniques were employed to synthesize the disclosed peptides. Fmoc (fluorenylmethyloxycarbonyl) chemistry occurred on PEG-base Rink Amide resin, 0.45mmol/g loading. Side-chain protection of Fmoc amino acid included t- Bu (t-butyl) on Asp, Ser, Tyr and D-Tyr; Trt (trityl) on Asn and Cys; Boc (tert- butyloxycarbonyl) on Lys; Pbf (2,2,4,6,7-r½ntamemyldihydroberizofurane) on Arg, Fmoc on K* (i.e., Fmoc-Lys(Fmoc)-OH was used to coupled to the resin).

General Procedure

1. Fmoc deprotection

[0037] Fmoc deprotection was performed using 20% piperidine in DMF

(dimethylformamide) or 20% piperidine / 0.1% 6-Cl HOBt (Hydroxybenzotriazole) in DMF, Smin + 20 min for each deprotection.

2. Coupling of Fmoc amino acids

[0038] Acylations were carried out using 3-fold Fmoc amino acids activated with DIC

(3eq.) (Ν,Ν'-Diisopropylcarbodiimide) in the presence of 6-Cl-HOBt (3eq.). Each coupling reaction took on average 1-3 h for completion. Kaiser test was used throughout for the in-process control of the coupling completion and the Fmoc deprotection. In the case that the Kaiser test was not satisfactory, re-couplings were done using DIC/6-C1 HOBt or HCTU (2 - (6 - Chloro - 1 H - benzotriazole - 1 - yl) - 1 , 1 ,3,3 - tetramethylaminium hexafluorophosphate)/OxymaPure.

3. Resin cleavage and Peptide Recovery

[0039] Peptidyl resin was extensively washed with DCM (dicloromethane) and methanol to remove the trace of DMF and dried at RT overnight. Peptidyl resin was clcavaged with the mixture of TFA/TIS/EDT/H₂0 (trifluoracctic acid/triethylsilane/1,2- ethanedithiol/water) (92.5/2.5/2.5/2.5, v/v/v/v) for 2.5-3 h under nitrogen bubbling.

Peptides were collected by precipitation in cold diethyl ether/Hexane (1/1, v/v).

4. Purification of Linear Peptide

[0040] Crude liner peptide was purified using Phenomenex Gemini C- 18 prep column. 21.5mm x 250mm. The peptide was then dried by lyophilization..

5. Disulfide Bridge Formation

[0041] Purified linear peptide was dissolved in 0.1M ammonium bicarbonate (3- Stng/ml), stand open to air for 2 days. Frequent HPLC testings were done to monitor the cyclization process. After cyclization, use TFA to adjust to acidic, isolated the product by lyophilization.

6. Cyclic Peptide Purification

[0042] Peptides were purified using the following conditions. Column: Phenomenex Gemini, 5u, CI 8, 1 10A, 250x21.2mm. Mobile phase A: 0.1% TFA in Acetonitrile/H₂0 (50/50). Mobile phase B: 0.1% TFA/H₂0. Gradient: 0-100% A in 120min. Wavelength: 229 nm. Flow rate: 16-20ml/min.

[0043] It will be appreciated mat the scope of this invention is to be defined by the appended claims rather than by the specific embodiments mat have been represented by way of example. The contents of all references (including literature references, issued patents, published patent applications, and co-pending patent applications) cited throughout this application are hereby expressly incorporated herein in their entireties by reference. Unless otherwise defined, all technical and scientific terms used herein are accorded the meaning commonly known to one with ordinary skill in the art.

Claims

CLAIMS:

1. A peptide of the Formula SEQ ID No. 1 :

or a pharmaceutically acceptable salt thereof, wherein

J¹ is Aha or Arg;

X¹ to X⁷ are each independently a natural or non-natural amino acid;

s, and t are each independently 0 or 1 ;

J² and J³ are each independently Lys or Arg;

J⁴, if present is Lys or Arg; and

J⁵, if present is Lys or Arg.

2. The peptide of Claim 1, wherein X¹ is selected from Arg, Lys, His, Pro, Cys, Thr, Ser, Gin, Glu, Leu, He, Met, Ala, Val, Gly, n-Leu, Met, Asp, and He.

3. The peptide of Claim 1 or 2, wherein X¹ is selected from Arg, Gin, Glu, Leu, n- Leu, Met, Asp, and He.

4. The peptide of any one of Claims 1 to 3, wherein X¹ is selected from Gin, Leu, and n-Leu.

5. The peptide of any one of Claims 1 to 4, wherein X² is selected from a natural amino acid.

6. The peptide of any one of Claims 1 to 5, wherein X² is selected from Ser, Thr, Cys, Met, Asn, Lys, Gin, His, Arg, Glu, and Asp.

7. The peptide of any one of Claims 1 to 6, wherein X² is selected from Ser, Gin, His, Arg, Glu, and Asp.

8. The peptide of any one of Claims I to 7, wherein X is selected from Gin and His.

9. The peptide of any one of Claims 1 to 8, wherein X is selected from a natural amino acid.

10. The peptide of any one of Claims 1 to 9, wherein X³ is selected from Tyr, Trp, Phe, Ser, Thr, Cys, Met, Asn, Ala, Lys, His, Glu, Arg, and Asp.

11. The peptide of any one of Claims 1 to 10, wherein X³ is selected from Tyr, Ser, Ala, Glu, Arg, and Asp.

12. The peptide of any one of Claims 1 to 1 1 , wherein X³ is selected from Tyr, Ser, Arg, and Asp.

13. The peptide of any one of Claims 1 to 12, wherein X⁴ is selected from a natural amino acid.

14. The peptide of any one of Claims 1 to 13, wherein X⁴ is selected from Gly, Ala, Val, Leu, He, Pro, Phe, Tyr, Trp, Asp, Glu, Lys, Arg, and His.

15. The peptide of any one of Claims 1 to 14, wherein X⁴ is selected from Leu, Asp, Gly, Tyr, Glu, and Arg.

16. The peptide of any one of Claims 1 to 15, wherein X⁴ is selected from Leu, Asp, Tyr, and Arg.

17. The peptide of any one of Claims 1 to 16, wherein X^s is selected from a natural amino acid.

18. The peptide of any one of Claims 1 to 17, wherein X^s is selected from Ser, Thr, Cys, Met, Asn, Gin, Asp, Glu, Lys, Arg, and His.

19. The peptide of any one of Claims I to 18, wherein X^s is selected from Tyr, Ser, Arg, Glu, and Asp.

20. The peptide of any one of Claims 1 to 19, wherein X⁶ is selected from Gly, Ala, Val, Leu, He, Pro, Phe, Tyr, Trp, Ser, Cys, Met, Gin, Lys, Arg, and His.

21. The peptide of any one of Claims 1 to 20, wherein X⁶ is selected from He, Leu, Pro, Val, Ala, Gly, His, Lys, and Arg.

22. The peptide of any one of Claims 1 to 21, wherein X⁶ is selected from He, Leu, and Arg.

23. The peptide of any one of Claims 1 to 22, wherein X⁶ is selected from He and Leu.

24. The peptide of any one of Claims 1 to 23, wherein X⁷ is selected from Gly, Ala, Val, Leu, lie, Pro, Phe, Tyr, Trp, Ser, Thr, Cys, Met, Asn, Gin, Lys, Arg, and His.

25. The peptide of any one of Claims 1 to 24, wherein X⁷ is selected from Tyr, Phe, His, Lys, Arg, and Trp.

26. The peptide of any one of Claims 1 to 25, wherein X⁷ is selected from Tyr, Arg, and Trp.

27. The peptide of any one of Claims 1 to 26, wherein X⁷ is Tyr.

28. The peptide of any one of Claims 1 to 27, wherein s and t are each 0.

29. The peptide of any one of Claims 1 to 28, wherein s and t are each 1.

30. The peptide of any one of Claims 1 to 29, where J¹ is Aha.

31. The peptide of Claim 1 , wherein the peptide is selected from

or a pharmaceutically acceptable salt thereof.

32. A pharmaceutical composition comprising a peptide of any one of Claims 1 to 31 , or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable carrier.

33. A method of treating irritable bowel disease in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a peptide of any one of Claims 1 to 32, or a pharmaceutically acceptable salt thereof.

34. The method of Claim 33, wherein the irritable bowel disease is Crohn's disease or ulcerative colitis.