Compositions and methods for treating endometriosis

Abstract

To provide non-hormonal, non-surgical treatment for endometriosis.SOLUTION: Provided herein are peptides that bind β-catenin, and compositions comprising the peptides. Peptides binding β-catenin prevent translocation to the nucleus and modulate the canonical Wnt pathway. Also provided herein are methods of using the peptides and compositions in the treatment of tissue-infiltrating conditions including endometriosis.SELECTED DRAWING: Figure 4C

Description

[Technical field]

[0001] cross reference This patent application claims the benefit of U.S. Provisional Patent Application No. 62 / 770,601, filed November 21, 2018, which is incorporated by reference in its entirety.

[0002] The present disclosure relates to agents and methods for treating disease or illness, formulations, and devices for administering such agents to a subject. Some of the agents provided can inhibit certain cellular proteins or functions, such as inhibitors of β-catenin and Wnt. The agents can include therapeutic peptides and peptidomimetics. The features of the methods provide various advantages for treating diseases such as endometriosis, such as the ability to treat or reverse the underlying causes of endometriosis, and to reduce or eliminate the need for hormone therapy to ameliorate symptoms associated with endometriosis. [Background technology]

[0003] Endometriosis (EMS) affects ~10% of women and adolescent females, affecting over 7.4 million women in the United States and over 176 million women worldwide. EMS is the leading cause of disability and infertility among women of reproductive age, with 40% of infertile women suffering from EMS. The average age at diagnosis is 28 years. Debilitating symptoms include painful menstruation, chronic pain, painful intercourse, and infertility (Giudice, LC & Kao, LC, Lancet 364, 1789-1799). In addition to human suffering, the productivity loss and direct medical costs associated with the condition in the United States exceed $90 billion annually. EMS results from retrograde menstruation, in which viable endometrial tissue flows backward up the fallopian tubes and into the peritoneal cavity. There, viable endometrial tissue is attached to multiple foreign sites (e.g., fallopian tubes, ovarian fossae, peritoneal wall, ligaments, and bowel) and responds to hormones (Sampson, JA, Am J Pathol 3,93-110 143,1927). Current treatments are only symptomatic and are not effective in curing. Recurrence is common and may occur quickly if treatment is discontinued.

[0004] Hormone-based treatments are used to alter ovulation cycles to reduce retrograde menstruation and treat EMS symptoms. These include estrogen / progesterone combination treatments, progestin-only treatments, or GnRH antagonists / agonists (i.e., "medical menopause"), which reduce hormonal activity and suppress ovulation; however, these treatments only suppress EMS symptoms but do not cure the disease. If treatment is discontinued, recurrence of symptoms is ~70% (Selcuk, I. et al., Ger Gynecol Assoc 14, 98-103, 2013). Management of EMS by hormonal modulation fails to reverse existing lesions; does not interfere with the development of lesions caused by other factors (e.g., oxidative stress or genetics); is hindered by progesterone receptor (PGR) resistance, which is seen in most women with EMS; and is associated with undesirable side effects. The limitations of hormone therapy for EMS result in limited efficacy, low adherence to protocols, and the need for invasive surgery.

[0005] Although surgery remains a fundamental tool in diagnosing and managing endometriosis, even surgery is not curative; the primary goal of surgical treatment is to prevent recurrence and relieve symptoms in order to obviate the need for or prolong the time until further surgery.

[0006] Therefore, there is a need for non-hormonal, non-surgical treatments for EMS. Summary of the Invention

[0007] Formula X1-X2-X3-X4-X5-X6-X7-X8-X9-X 10 -X 11 -X 12 -X 13 -X 14 -X 15 -X 16Provided herein are peptides and peptidomimetics comprising an amino acid sequence having the formula: wherein: X1 is M or null; X2 is S, I, G, T, A, L, or null; X3 is R, K, or null; X4 is a positively charged amino acid, citrulline, Orn, D, E, 8-aminooctanoic acid, or an amino carboxylic acid having 4-12 carbons; X5 is M, norleucine, Orn, D, E, K, H, R, K, 8-aminooctanoic acid, an amino carboxylic acid having 4-12 carbons, or null; X6 is W, Y, F, or N-methyl A; X7 is F, I, L, Chg, Cha, or Tle; X8 is L, I, or A; X9 is L, I, or A; 10 is C, S, A, Abu, C(me), or S(Bzl); 11 is F, H, A, K, E, Chg, Cng, or Orn; X 12 is W, Y, A, or F; and X 13 is G, GABA, or null.

[0008] Formula R-X1-X2-X3-X4-X5-X6-X7-X8-X9-X 10 -X 11 -X 12 -X 13 -X 14 -X 15 -X 16 Provided herein are peptides and peptidomimetics comprising an amino acid sequence having the formula: X3 is P, W, N-methyl-W, N-ethyl-W, N-methyl A, N-ethyl A, L, Pip, Aib, Y, or F; X4 is E, Q, N, or D; X5 is S, alpha methyl S, K, D, Orn, T, or E; X6 is I, Chg, H, or L; X7 is L or I; X8 is D, N, E, or Q; X9 is D, E, K, Q, or Orn; X10 is H or methyl-H; X 11 is V, α-methyl V, Chg, L, I, or norvaline; X 12 is Q, Aib, S, R, or N; X 13 is R, K, Citrulline, Orn, D, or E; X 14 is V, I, L, or norvaline; X 15 is W, Y, or F; and X 16 is R, G, or null.

[0009] Provided herein are peptides and peptidomimetics comprising the amino acid sequence of any one of SEQ ID NO:1 to SEQ ID NO:500.

[0010] Provided herein are peptides and peptidomimetics comprising an amino acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to any one of SEQ ID NO:1 through SEQ ID NO:500.

[0011] In some embodiments, the peptide binds to β-catenin. In some embodiments, the peptide is a β-catenin inhibitor. In some embodiments, the peptide is an inhibitor of β-catenin translocation to the nucleus. In some embodiments, the peptide prevents β-catenin from acting as a transcription factor on oncogenes, matrix metalloproteinase 9 (MMP9), or chloride C3 channel (ClC-3). In some embodiments, the peptide prevents endometriosis (EMS) cell transformation, invasion, migration, fibrosis, or any combination thereof. In some embodiments, the peptide prevents β-catenin from binding to estrogen receptor (ESR1). In some embodiments, the peptide does not reduce β-catenin membrane activity. In some embodiments, the peptide does not reduce β-catenin E-cadherin binding. In some embodiments, the peptide prevents oncogenic transcription factor activity. In some embodiments, the peptides are at a concentration of less than 50 uM, less than 30 uM, less than 10 uM, 5 uM, 1 uM, 500 nM, 400 nM, 300 nM, 200 nM, 100 nM, 50 nM, 30 nM, 10 nM, 5 nM, 3 nM, 1 nM, 800 pM, 600 pM, 400 pM, 200 pM, 100 pM, 50 pM, 30 pM, 20 pM, 10 pM, or 5 pM, or about 10 uM. In one embodiment, the compound inhibits Wnt pathway activity with an EC50 of less than about 5 uM, about 1 uM, about 500 nM, about 400 nM, about 300 nM, about 200 nM, about 100 nM, about 50 nM, about 30 nM, about 10 nM, about 5 nM, about 3 nM, about 1 nM, about 800 pM, about 600 pM, about 400 pM, about 200 pM, about 100 pM, about 50 pM, about 30 pM, about 20 pM, about 10 pM, or about 5 pM.In some embodiments, the peptide is at a concentration of less than 50 uM, 30 uM, 10 uM KD, 5 uM, 1 uM, 500 nM, 400 nM, 300 nM, 200 nM, 100 nM, 50 nM, 30 nM, 10 nM, 5 nM, 3 nM, 1 nM, 800 pM, 600 pM, 400 pM, 200 pM, 100 pM, 50 pM, 30 pM, 20 pM, 10 pM, or 5 pM, or less than about 10 uM. , about 5 uM, about 1 uM, about 500 nM, about 400 nM, about 300 nM, about 200 nM, about 100 nM, about 50 nM, about 30 nM, about 10 nM, about 5 nM, about 3 nM, about 1 nM, about 800 pM, about 600 pM, about 400 pM, about 200 pM, about 100 pM, about 50 pM, about 30 pM, about 20 pM, about 10 pM, or about 5 pM. In some embodiments, the peptide is not naturally occurring. In some embodiments, the peptide is a cyclized peptide. In some embodiments, the peptide is a bicyclic peptide. In some embodiments, the peptide is cyclized at a Cys-Cys disulfide bond. In some embodiments, the peptide is cyclized at an amide bond. In some embodiments, the amide bond is head-to-tail between the N-terminus and the C-terminus. In some embodiments, the amide bond is head-to-side chain between the N-terminus and an internal COOH. In some embodiments, the amide bond is chain-to-tail between an internal NH2 and the C-terminus. In some embodiments, the amide bond is side chain-to-side chain between an internal NH2 and an internal COOH. In some embodiments, the peptide is cyclized using hydrocarbon stapling. In some embodiments, the peptide is cyclized using click chemistry.In some embodiments, the peptide is at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, at least 21, at least 22, at least 23, at least 24, at least 25, at least 26, at least 27, at least 28, at least 29, at least 30, at least 31, at least 32, at least 33, at least 34, at least 35, at least 36, at least 37, at least 38, at least 39, at least 40, at least 41, at least 42, at least 43, at least 44, at least 45, at least 46, at least 47, at least 48, at least 49, at least 50, at least 51, at least 52, at least 53, at least 54, at least 55, at least 56, at least 57, at least 58, at least 59, at least 60, at least 61, at least 62, at least 63, at least 64, at least 65, at least 66, at least 67, at least 68, at least 69, at least 70, at least 71, at least 72, at least 73, at least 74, at least 75, at least 76, at least 77, at least 78, at least 79, at least 80, or at least 81 amino acid residues.In some embodiments, the peptide is less than 4, less than 5, less than 6, less than 7, less than 8, less than 9, less than 10, less than 11, less than 12, less than 13, less than 14, less than 15, less than 16, less than 17, less than 18, less than 19, less than 20, less than 21, less than 22, less than 23, less than 24, less than 25, less than 26, less than 27, less than 28, less than 29, less than 30, less than 31, less than 32, less than 33, less than 34, less than 35, less than 36, less than 37, less than 38, less than 39, less than 40, less than 41, less than 42, less than 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, or 81 amino acid residues. In some embodiments, the peptide comprises one or more unnatural amino acids. In some embodiments, the one or more unnatural amino acids are N-methyl amino acids.

[0012] Provided herein is a pharmaceutical composition comprising a β-catenin inhibitor and a pharma- ceutically acceptable carrier.

[0013] Provided herein is a pharmaceutical composition comprising a peptide that comprises binding specificity for β-catenin and a pharma- ceutically acceptable carrier.

[0014] Provided herein are pharmaceutical compositions comprising a β-catenin inhibitor, including a peptide, and a pharma- ceutically acceptable carrier.

[0015] Provided herein is a pharmaceutical composition comprising a peptide as described herein; and a pharma- ceutically acceptable carrier. In some embodiments, the peptide is a cyclized peptide. In some embodiments, the pharmaceutical composition comprises an absorption enhancer or permeation enhancer. In some embodiments, the absorption enhancer is one or more sulfoxides, such as dimethyl sulfoxide (DMSO); laurocapram (1-dodecylazacycloheptan-2-one); pyrrolidones, such as n-methyl-2-pyrrolidone; terpenes and terpenoids; essential oils; oxazolidinones, such as 4-decycloxazolidin-2-one; urea; cyclopentadecalactone; sodium N-[8-(2-hydroxybenzoyl)amino]caprylate (SNAC); 8-(N-2-hydroxy-5-chloro-benzoyl)-amino-caprylic acid (5-CNAC); medium chain fatty acids, salts, and derivatives; sodium caprate (sodium caprate). caprate); sodium caprylate; protease inhibitors and omega-3 fatty acids; liquid mixed micellar propellants; lipid polymer micelles; alkyl glycosides; chitosan; dodecyl-2-N,N-dimethylaminopropionate (DDAIP); cell membrane lipid components; nanoparticles; liposomes, ligands; and lipophilic modifications. In some embodiments, the absorption enhancer is sodium caprate.

[0016] Formulations comprising the peptides or pharmaceutical compositions described herein are provided herein, wherein the formulations comprise solutions, lotions, concocts, creams, ointments, gels, foams, powders, solids, pastes, tinctures, microparticles, microcapsules, nanoparticles, liposomes, emulsions, or lyophilisates. Formulations are further described herein, wherein the gel is a thermoreversible gel. Formulations are further described herein, wherein the formulations comprise one or more mucoadhesive polymers. Formulations are further described herein, wherein the formulations are for topical administration.

[0017] Provided herein is an intravaginal device comprising the peptide or pharmaceutical composition described herein. In some embodiments, the intravaginal device is a suppository, a transdermal patch, a sponge, a tape, a film, an intravaginal ring, a vaginal tampon, a vaginal ring, a vaginal strip, a vaginal capsule, a vaginal tablet, a vaginal pessary, a vaginal cup, a vaginal sponge, or an intrauterine device. In some embodiments, the peptide or pharmaceutical composition is formulated as a formulation selected from the group consisting of a solution, a lotion, a concomitant, a cream, an ointment, a gel, a foam, a mucoadhesive composition, a coating, a core, a matrix, an emulsion, a liposome, or a lyophilisate, wherein the intravaginal device comprises the formulation. In some embodiments, the intravaginal device comprises about 0.01 mg to about 5000 mg of the inhibitor. In some embodiments, the intravaginal device comprises about 0.01 mg, about 0.05 mg, about 0.1 mg, about 0.5 mg, about 1 mg, about 5 mg, about 10 mg, about 20 mg, about 40 mg, about 60 mg, about 80 mg, about 100 mg, about 150 mg, about 200 mg, about 400 mg, about 600 mg, about 800 mg, about 1000 mg, about 1200 mg, about 1400 mg, about 1600 mg, about 1800 mg, about 2000 mg, about 2500 mg, about 3000 mg, about 3500 mg, about 4000 mg, about 4500 mg, or about 5000 mg of the inhibitor. In some embodiments, the intravaginal device is configured to deliver the inhibitor transmucosally. In some embodiments, the intravaginal device is configured to detect and treat the following: 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, 24 hours, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks ... In some embodiments, the inhibitor is configured to deliver the inhibitor for one or more of the following: weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, 1 year, 2 years, 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, 11 years, or 12 years.In some embodiments, the intravaginal device is configured to deliver about 0.01 mg to about 1000 mg of the inhibitor per day. In some embodiments, the intravaginal device is configured to deliver about 0.01 mg, about 0.05 mg, about 0.1 mg, about 0.5 mg, about 1 mg, about 5 mg, about 10 mg, about 20 mg, about 40 mg, about 60 mg, about 80 mg, about 100 mg, about 150 mg, about 200 mg, about 400 mg, about 600 mg, about 800 mg, or about 1000 mg of the inhibitor per day. In some embodiments, the device is a suppository. In some embodiments, the device is a transdermal patch. In some embodiments, the device is a sponge. In some embodiments, the device is a tape. Further provided herein is an intravaginal device, wherein the device is an intravaginal ring. In some embodiments, the intravaginal ring comprises a silicone insert, a compressed tablet, or a lyophilized gel. In some embodiments, the peptide or composition is incorporated throughout a silicone insert, compressed tablet, or lyophilized gel. In some embodiments, the device is a vaginal tampon. In some embodiments, the device is a vaginal ring. In some embodiments, the device is a vaginal strip. In some embodiments, the device is a capsule. In some embodiments, the device is a vaginal tablet. In some embodiments, the device is a vaginal pessary. In some embodiments, the device is a vaginal cup. In some embodiments, the device is a vaginal sponge. In some embodiments, the device is an intrauterine device.

[0018] Provided herein is a formulation comprising the peptide or pharmaceutical composition described herein, wherein the formulation is for oral administration.Further provided herein is a formulation, wherein the formulation is a tablet or capsule.In some embodiments, the formulation is liquid.

[0019] Provided herein is a method of inhibiting β-catenin in a subject, comprising inserting a device described herein into the vagina of the subject.

[0020] Provided herein are methods of inhibiting β-catenin comprising contacting a cell with a peptide or pharmaceutical composition described herein.

[0021] Provided herein is a method of treating endometriosis, comprising administering to a subject in need thereof a therapeutically effective amount of a peptide or pharmaceutical composition described herein.

[0022] Provided herein is a method for reducing symptoms associated with endometriosis, comprising administering a therapeutically effective amount of a peptide or pharmaceutical composition described herein to a subject in need thereof. In some embodiments, the symptoms include at least one selected from the group consisting of chronic pain, central sensitization, myofascial pain, adnexal mass, infertility, dysmenorrhea, genetic predisposition, non-menstrual pelvic-abdominal pain, dyspareunia, bowel symptoms (diarrhea, cramps, constipation), painful defecation (difficulty in passing stool), ovarian mass or tumor, painful bladder symptoms, and dysuria.

[0023] Provided herein is a method of treating a disease comprising administering to a subject a therapeutically effective amount of a peptide or pharmaceutical composition described herein. In some embodiments, the disease is a tissue-infiltration condition, a tissue migration condition, a tissue invasion condition, or a combination thereof. In some embodiments, the disease comprises cancer. In some embodiments, the cancer comprises colorectal cancer, squamous cell carcinoma, head and neck cancer, pancreatic cancer, breast cancer, myeloid leukemia, basal cell carcinoma, synovial sarcoma, non-small cell lung cancer, solid tumors, or prostate cancer. In some embodiments, the disease comprises endometriosis. In some embodiments, the peptide binds to β-catenin. In some embodiments, the peptide inhibits β-catenin. In some embodiments, the peptide binds to cytoplasmic β-catenin. In some embodiments, the peptide binds to cytoplasmic β-catenin and inhibits translocation of β-catenin to the nucleus of the cell. In some embodiments, the peptide binds to cytoplasmic β-catenin and maintains or increases membrane-bound β-catenin. In some embodiments, the peptide binds to cytoplasmic β-catenin and reduces nuclear β-catenin. In some embodiments, the peptide binds to cytoplasmic β-catenin and prevents β-catenin from acting as a transcription factor on oncogenes, matrix metalloproteinase 9 (MMP9), or chloride C3 channel (ClC-3). In some embodiments, the peptide binds to cytoplasmic β-catenin and prevents EMS cell transformation, invasion, migration, fibrosis, or any combination thereof. In some embodiments, the peptide binds to cytoplasmic β-catenin and prevents β-catenin from binding to estrogen receptor (ESR1). In some embodiments, the peptide binds to cytoplasmic β-catenin and does not reduce β-catenin membrane activity. In some embodiments, the peptide binds to cytoplasmic β-catenin and does not reduce β-catenin-E-cadherin binding. In some embodiments, the peptide prevents oncogenic transcription factor activity. In some embodiments, the decrease in free cytoplasmic β-catenin is for cells of a subject obtained at different time points.In some embodiments, the amount of membrane-bound cytoplasmic β-catenin in the cells of the subject is increased by at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 100%. In some embodiments, the increase in membrane-bound cytoplasmic β-catenin is relative to cells of a control subject that has not been administered a therapeutically effective amount of the pharmaceutical composition. In some embodiments, the increase in membrane-bound β-catenin is relative to cells of the subject obtained before the subject develops the disease. In some embodiments, the increase in membrane-bound β-catenin is relative to cells of the subject obtained at a different time point. In some embodiments, the amount of nuclear β-catenin in the cells of the subject is reduced by at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 100%. In some embodiments, the reduction in nuclear β-catenin is relative to cells of a control subject that has not been administered a therapeutically effective amount of the pharmaceutical composition. In some embodiments, the reduction in nuclear β-catenin is relative to cells of the subject obtained before the subject develops the disease. In some embodiments, the reduction in nuclear β-catenin is relative to cells of the subject obtained at a different time point.

[0024] In some embodiments, the therapeutically effective amount is from about 0.01 mg to about 1000 mg.

[0025] Provided herein is a method of preventing a disease comprising administering to a subject a therapeutically effective amount of a peptide or pharmaceutical composition described herein. In some embodiments, the disease is a tissue invasion disease, a tissue migration disease, a tissue invasion disease, or a combination thereof. In some embodiments, the disease comprises cancer. In some embodiments, the cancer comprises colorectal cancer, squamous cell carcinoma, head and neck cancer, pancreatic cancer, breast cancer, myeloid leukemia, basal cell carcinoma, synovial sarcoma, non-small cell lung cancer, solid tumors, or prostate cancer. In some embodiments, the disease comprises endometriosis. In some embodiments, the peptide binds to β-catenin. In some embodiments, the peptide inhibits β-catenin. In some embodiments, the peptide binds to cytoplasmic β-catenin. In some embodiments, the peptide binds to cytoplasmic β-catenin and inhibits the translocation of β-catenin to the nucleus of the cell. In some embodiments, the peptide binds to cytoplasmic β-catenin and maintains or increases the amount of membrane-bound β-catenin. In some embodiments, the peptide binds to cytoplasmic β-catenin and prevents β-catenin from acting as a transcription factor on oncogenes, matrix metalloproteinase 9 (MMP9), or chloride C3 channel (ClC-3). In some embodiments, the peptide prevents EMS cell transformation, invasion, migration, fibrosis, or any combination thereof. In some embodiments, the peptide prevents β-catenin from binding to estrogen receptor (ESR1). In some embodiments, the peptide binds to cytoplasmic β-catenin and the membrane activity of β-catenin is not reduced. In some embodiments, the peptide binds to cytoplasmic β-catenin and β-catenin-E-cadherin binding is unaffected. In some embodiments, the peptide prevents oncogenic transcription factor activity.In some embodiments, the amount of nuclear β-catenin in the cells of the subject is reduced by at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 100%. In some embodiments, the reduction in nuclear β-catenin is relative to cells of a control subject that has not been administered a therapeutically effective amount of the pharmaceutical composition. In some embodiments, the reduction in nuclear β-catenin is relative to cells of the subject obtained before the subject develops endometriosis or a tissue-infiltrating condition. In some embodiments, the reduction in nuclear β-catenin is relative to cells of the subject obtained at a different time point. In some embodiments, the therapeutically effective amount is about 0.01 mg to about 1000 mg. In some embodiments, the peptide or pharmaceutical composition is administered intravenously. In some embodiments, the peptide or pharmaceutical composition is administered intramuscularly. In some embodiments, the peptide or pharmaceutical composition is administered simultaneously with administration of a drug for treating osteoporosis. In some embodiments, the drug for treating osteoporosis comprises alendronate, ibandronate, risedronate, zoledronic acid, denosumab, calcitonin, estrogen, raloxifene, bazodoxifene, teriparatide, abaloparatide, or any combination thereof. In some embodiments, the drug for treating osteoporosis is zoledronic acid.

[0026] Provided herein is an intravaginal device comprising a peptide comprising the amino acid sequence of SEQ ID NO: 215 or 393 formulated to release the peptide in a time-controlled manner, wherein the peptide binds to β-catenin, wherein the peptide inhibits the translocation of β-catenin to the nucleus of a cell, wherein the peptide does not reduce β-catenin binding to E-cadherin in the membrane, and wherein the peptide reduces the size and severity of endometriotic lesions or other symptoms associated with endometriosis in a subject.

[0027] INCORPORATION BY REFERENCE All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference. [Brief description of the drawings]

[0028] [Figure 1] The "Off" and "On" states of the Wnt / β-catenin signaling pathway are shown. [Figure 2A] FIG. 1 shows luciferase expression in transfected reporter cells controlled by a Wnt-responsive promoter incubated with a β-catenin inhibitory peptide. [Figure 2B] FIG. 1 shows cell proliferation in a β-catenin expressing cell line following incubation with β-catenin inhibitory peptides (SEQ ID NO:264 and SEQ ID NO:396). [Diagram 3] 1 shows E-cadherin binding in the presence of a β-catenin inhibitory peptide corresponding to SEQ ID NO:42. [Figure 4A] Figure 4A shows that peptide inhibitors against β-catenin cause regression of lesions.Figure 4A shows analysis of daily vaginal smears to assess estrous cycle stage. [Figure 4B] Figure 4B shows that peptide inhibitors against β-catenin cause lesion regression. Figure 4B shows that peptide 2 (SEQ ID NO: 215) demonstrates lesion regression. [Figure 4C] Figure 4C shows that peptide inhibitors against β-catenin cause regression of the lesions. Figure 4C shows images of lesions treated with either vehicle or peptide 2. Two images of the lesions (L) are shown for each treatment. The donor mice express green fluorescent protein and the lesions are green. Magnification = 7.5x. [Diagram 5] A shows the histology of a sample obtained from the EMS lesion of a mouse treated with vehicle alone, and B shows the histology of a sample obtained from the EMS lesion of a mouse treated with peptide 2. [Figure 6A] Representative slides at 20x magnification of endometriotic lesions from a mouse model stained to show that β-catenin is not localized to the nucleus after treatment with therapeutic peptides and that peptide inhibitors follow the b-catenin localization pattern are shown. [Figure 6B] A representative slide at 120x magnification of stained endometriotic lesions from a mouse model is shown, showing greater localization of β-catenin to the membrane in peptide-treated samples compared to control-treated mice. [Figure 7] A compares membrane-bound β-catenin quantification between lesions treated with therapeutic peptides and untreated controls, and B compares nuclear β-catenin quantification between lesions treated with therapeutic peptides and untreated controls. [Figure 8A] 1 shows that the therapeutic peptide inhibits Wnt pathway gene transcription to a greater extent than a known positive control (ICG001). [Figure 8B] 1 shows that the therapeutic peptide inhibits Wnt pathway gene transcription to a greater extent than a known positive control (ICG001). [Figure 9] 1 shows the results of a fluorescence polarization assay using a peptide having SEQ ID NO:404. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0029] For example, drugs, compositions, methods, and products are provided herein for use in conjunction with treatment therapies for the treatment of various diseases and disorders. In some cases, the diseases and disorders may include endometriosis. The methods may include administering to a subject an inhibitor, or a composition, formulation, or device that includes the inhibitor. Exemplary inhibitors include β-catenin inhibitors. The inhibitors may include peptides, such as isolated peptides or combinations of peptides.

[0030] I. Definition Unless otherwise defined, all technical terms, notations, and other technical and scientific or terminology used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the claimed subject matter belongs. In some cases, terms with commonly understood meanings are defined herein for clarity and / or ready reference, and the inclusion of such definitions herein should not necessarily be construed as representing a substantial difference to what is commonly understood in the art.

[0031] Throughout this application, various embodiments may be presented in a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the present disclosure. Thus, the description of a range should be considered to have specifically disclosed all possible subranges as well as individual numerical values ​​within that range. For example, the description of a range such as 1-6 should be considered to have specifically disclosed subranges such as 1-3, 1-4, 1-5, 2-4, 2-6, 3-6, as well as individual numerical values ​​within the range, such as 1, 2, 3, 4, 5, and 6. This applies regardless of the breadth of the range.

[0032] As used in this specification and claims, "a," "and," and "the" include plural references unless the context clearly dictates otherwise. For example, the term "a sample" includes a plurality of samples, including mixtures thereof.

[0033] "Determining," "measuring," "evaluating," "assessing," "assaying," and "analyzing" are often used interchangeably herein to refer to forms of measurement and include determining the presence or absence of an element (e.g., detection). Such terms can include quantitative, qualitative, or both quantitative and qualitative determinations. Evaluation is alternatively relative or absolute. "Detecting the presence of" includes determining the amount of something present, as well as determining its presence or absence.

[0034] The terms "subject," "individual," or "patient" are often used interchangeably herein. A "subject" can be a biological entity that contains expressed genetic material. The biological entity can be, for example, a plant, an animal, or a microorganism, including bacteria, viruses, fungi, and protozoa. A subject can be tissues, cells, and their progeny of a biological entity obtained in vivo or cultured in vitro. A subject can be a mammal. A mammal can be a human. A subject can be diagnosed or suspected of being at high risk for a disease. The disease can be endometriosis. In some cases, a subject is not necessarily diagnosed or suspected of being at high risk for a disease.

[0035] The term "in vivo" is used to describe events that take place in a subject's body.

[0036] The term "in vitro" is used to describe events that occur outside of a subject's body. An "ex vivo" assay is not performed on a subject. Rather, it is performed on a sample separate from the subject. An example of an "ex vivo" assay that is performed on a sample is an "in vitro" assay.

[0037] The term "in vitro" is used to describe events that occur in a container for holding a laboratory reagent that separates the laboratory reagent from the living organism of the biological source from which the material is obtained. In vitro assays can include cell-based assays in which living or dead cells are utilized. In vitro assays can also include cell-free assays in which no intact cells are utilized.

[0038] As used herein, a number followed by the term "about" refers to a number that is plus or minus 10% of that number. A range followed by the term "about" refers to a range of minus 10% of the minimum value and plus 10% of the maximum value.

[0039] As used herein, the terms "treatment" or "treating" are used in reference to a pharmaceutical or other intervention regimen to obtain a beneficial or desired result in a recipient. A beneficial or desired result includes, but is not limited to, a therapeutic benefit and / or a prophylactic benefit. A therapeutic benefit may refer to the eradication or amelioration of the condition or underlying disorder being treated. A therapeutic benefit may also be achieved by eradicating or ameliorating one or more of the physiological symptoms associated with the underlying disorder such that an improvement is observed in the subject, even though the subject may still be affected by the underlying disorder. A prophylactic benefit includes delaying, preventing, or eliminating the appearance of a disease or condition, delaying or eliminating the onset of symptoms of a disease or condition, slowing, halting, or reversing the progression of a disease or condition, or any combination thereof. With respect to a prophylactic benefit, a subject who is at risk of developing a particular disease or who reports one or more physiological symptoms of a disease may receive treatment even if a diagnosis of the disease has not been made.

[0040] Abbreviations for many of the terms used in the sequences used herein are defined in Table 1.

[0041] [Table 1-1]

[0042] [Table 1-2]

[0043] The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described.

[0044] II. WNT canonical pathway / β-catenin The Wnt pathway is involved in the control of gene expression, cell behavior, cell adhesion, and cell polarity. The classical (β-catenin-dependent) Wnt signaling pathway is the best studied of the Wnt pathways and is highly conserved throughout evolution. In this pathway, Wnt signaling inhibits the degradation of β-catenin, which can regulate the transcription of many genes. Wnt signaling is activated by ligation of Wnt proteins to their respective dimeric cell surface receptors, composed of seven transmembrane frizzled proteins and LRP5 / 6. Upon ligation to their receptors, the cytoplasmic protein disheveled (Dvl) is recruited, phosphorylated, and activated. Activation of Dvl induces the dissociation of GSK-3β from Axin, causing the inhibition of GSK-3β. Then, as a result of the inactivation of the "degradation complex," the phosphorylation and degradation of β-catenin is inhibited. Stable β-catenin then translocates to the nucleus, causing changes in various target gene expression.

[0045] In normal cells, β-catenin typically binds to E-cadherin at the cell membrane and is regulated by the Wnt pathway, as shown in Figure 1. In normal gynecological epithelia, high concentrations of β-catenin are often found at the membrane, and low concentrations of β-catenin are found in the cytoplasm and nucleus (Aberle, H., et al., EMBO J 16, 3797-3804, 1997; Jha, RK, et al., FEBS Lett 580, 5653-5660, 2006; Lee, KY, et al., Trends Endocrinol Metab 18, 234-239, 2007; Jeong, JW et al., Oncogene 28, 31-40, 2009, each of which is incorporated herein by reference for disclosure). The Wnt / β-catenin signaling pathway is turned "off" and β-catenin interacts with the degradation complex, which phosphorylates and targets β-catenin for proteasomal degradation.

[0046] Conversely, in Wnt pathway dysregulation, β-catenin often dissociates from the membrane and accumulates in the cytoplasm. Once accumulated in the cytoplasm, β-catenin can often translocate to the nucleus, where it can function as a transcription factor for oncogenes (Clevers, H., Cell 127, 469-480, 2006), matrix metalloproteinase 9 (MMP9), chloride 3 channel (ClC-3), and other Wnt pathway activating proteins. These proteins are believed to be able to initiate the transformation, invasion, migration, and fibrosis of EMS cells (Guan,YTet al.,Hum Reprod 31,986-998,2016)(Matsuzaki,S.et al.,Mol Cell Ther 2,36,2014)(Zhang,L.et al.,Biol Reprod 94,70,2016)(Zhang,L.et al.,Mol Hum Reprod 22,526-535,2016). When the transduction pathway is in the "on" state, cytoplasmic β-catenin accumulates, translocates into the nucleus, binds to transcription factors (including ER-α), and stimulates the transcription of WNT target genes, e.g., ClC3, MMP9, and SOX.

[0047] In EMS, β-catenin is often detached from the membrane, either by receptor dysregulation or somatic mutation, and is thought to bind to the estrogen receptor (ESR1) and translocate into the nucleus, where it activates onco / endogenes and causes pathology (Valentijn, AJ et al., Hum Reprod 28, 2695-2708, 2013) (Kouzmenko, AP et al., J Biol Chem 279, 40255-40258, 2004).

[0048] Maintenance of β-catenin membrane activity is thought to be a hallmark of normal cells; Wnt antagonists that disrupt the membrane binding of E-cadherin and β-catenin are thought to increase cytoplasmic β-catenin concentrations and cell motility, which may result in undesirable proto-oncogene-like effects (Onder, TT et al., Cancer Res 68, 3645-3654, 2008). Therefore, Wnt antagonists specific for cytoplasmic β-catenin, which can inhibit β-catenin nuclear translocation and oncogenic transcription factor activity, represent an attractive approach for the treatment of EMS (Matsuzaki, S. et al., Mol Cell Ther 2, 36, 2014; Xiong, W. et al., Reproduction 150, 507-516, 2015; Pazhohan, A. et al., Eur J Obstet Gynecol Reprod Biol 220, 1-5, 2018, each of which is incorporated by reference herein for such disclosures).

[0049] Dysregulation of the Wnt pathway appears to be present in all lesion subtypes of EMS. For example, cytoplasmic β-catenin translocates to the nucleus to activate target genes. Preventing β-catenin from translocating to the nucleus targets downstream initiators of EMS without causing unwanted side effects from targeting upstream receptors. Wnt antagonists can reverse endometriotic lesions. To date, four Wnt antagonists (PKF 115-584, CGP049090, niclosamide, and β-catenin siRNA) have been successful in reversing EMS lesion progression across EMS subtypes; none have progressed to clinical trials due to off-target effects. A therapeutic agent that inhibits excess cytoplasmic β-catenin translocation and inhibits its transcription factor activity without interfering with β-catenin:E-cadherin interactions, preserving membrane protein function, would be a significant improvement over therapies currently contemplated in the art.

[0050] Normal uterine function typically involves an inactivated canonical Wnt pathway and β-catenin primarily associated with the E-cadherin membrane complex (Figure 1). Hormonal activation of the Wnt pathway releases β-catenin into the cytoplasm and translocates to the nucleus, which activates its nuclear transcriptional activity and leads to EMS pathogenesis.

[0051] A. β-catenin inhibitors In some embodiments, the compositions provided herein include a β-catenin inhibitor. β-catenin inhibitors may be useful in a variety of therapeutic settings, including, for example, treating, inhibiting, preventing, or reducing EMS and its associated symptoms. Such inhibitors may directly address EMS pathogenesis, reverse disease progression, and prevent or slow the recurrence of EMS. In some cases, β-catenin inhibitors can shrink existing EMS lesions. In some cases, β-catenin inhibitors can inhibit dysregulation of the Wnt pathway, which is believed to be one of the underlying causes of EMS. β-catenin inhibitors may limit EMS pathogenesis by preventing endometrial cell invasiveness. Alternatively or in addition, β-catenin inhibitors can suppress epithelial-mesenchymal transition (EMT). This suppression can limit the growth of endometrial lesions. Alternatively or in addition, β-catenin inhibitors may also reduce the paracrine production of TGF-β1 and Wnt1, which can reduce fibrosis in ovarian endometriomas. In addition, β-catenin inhibitors may be able to reduce MMP9 expression, which may result in reduced angiogenesis. Inhibition or reduction of angiogenesis may inhibit the ability of endometrial cells to proliferate outside the uterus.

[0052] As an exemplary advantage, targeting β-catenin can reduce the undesirable side effects associated with the targeting upstream receptor. In some cases, these side effects can include excessive estrogen production. In some cases, the inhibitor preferentially inhibits the ability of β-catenin to translocate from the cytoplasm to the nucleus. In some cases, the inhibitor preferentially inhibits the transcriptional activity of β-catenin. These exemplary features can advantageously leave the membrane function of β-catenin largely or completely intact, which in some cases can reduce the side effects associated with the inhibition of Wnt function. Thus, in some cases, the inhibitor does not inhibit the binding of β-catenin to the cell membrane.

[0053] In some cases, the inhibitor does not prevent β-catenin from binding to binding factor, which may cause side effects.For example, in some embodiments, the inhibitor may not prevent the binding or interaction between β-catenin and E-cadherin or adenomatous polyposis coli (APC), or may not prevent nuclear TCF from binding to DNA.

[0054] 1. Peptides In some embodiments, the β-catenin inhibitor comprises a peptide. In some cases, the peptide corresponds to, e.g., is based on or derived from, a protein or peptide that binds to β-catenin. In some cases, the peptide contains a synthetic peptide sequence.

[0055] The peptides described herein may be optimized for their affinity to their desired target, absorption or cellular uptake, stability, protease resistance, and other factors. The affinity of a peptide to its intended target can be modified using a variety of methods. For example, affinity can be improved using amino acid substitutions. Additionally, changing the size of the peptide, changing the position of cyclization, increasing hydrophobicity, and modifying amino acids or incorporating artificial amino acids into the structure of the peptide may affect the affinity of the peptide. Exemplary modifications include incorporating N-methyl and alpha-methyl amino acids.

[0056] Among the embodiments provided are peptidomimetics that contain the same or similar structure and properties as the peptides described herein.Such peptidomimetics include small protein-like chains that are designed to mimic peptides.These include peptide mimetics that include peptide modifications or similar system designs that mimic peptides, such as peptoids and β-peptides.

[0057] As depicted in Table 4, exemplary peptides include SEQ ID NO:1 through SEQ ID NO:500. The peptides are typically identified in one of two families: those with homology to the TCF4 / β-catenin binding region and those with sequences similar to the β-catenin binding staple peptides. In some cases, the peptides include an amino acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to any one of SEQ ID NO:1 through SEQ ID NO:500.

[0058] Alternatively or additionally, the peptide may have the formula X1-X2-X3-X4-X5-X6-X7-X8-X9-X 10 -X 11 -X 12 -X 13 -X 14 -X 15 -X 16In some embodiments, X1 is M or null. In some embodiments, X2 is S, I, G, T, A, L, or null; and X3 is R, K, or null. In some embodiments, X4 is a positively charged amino acid, citrulline, Orn, D, E, 8-aminooctanoic acid, or an amino carboxylic acid having 4-12 carbons. In some embodiments, X5 is M, norleucine, Orn, D, E, K, H, R, K, 8-aminooctanoic acid, an amino carboxylic acid having 4-12 carbons, or null. In some embodiments, X6 is W, Y, F, or N-methyl A. In some embodiments, X7 is F, I, L, Chg, Cha, or Tle. In some embodiments, X8 is L, I, or A. In some embodiments, X9 is L, I, or A. In some embodiments, X 10 is C, S, A, Abu, C(me), or S(Bzl). 11 is F, H, A, K, E, Chg, Cng, or Orn. 12 is W, Y, A, or F. In some embodiments, X 13 is G, GABA, or null. In some embodiments, X1 is M or null; X2 is S, I, G, T, A, L, or null; X3 is R, K, or null; X4 is a positively charged amino acid, citrulline, Orn, D, E, 8-aminooctanoic acid, or an amino carboxylic acid having 4-12 carbons; X5 is M, norleucine, Orn, D, E, K, H, R, K, 8-aminooctanoic acid, an amino carboxylic acid having 4-12 carbons, or null; X6 is W, Y, F, or N-methyl A; X7 is F, I, L, Chg, Cha, or Tle; X8 is L, I, or A; X9 is L, I, or A; 10 is C, S, A, Abu, C(me), or S(Bzl); 11 is F, H, A, K, E, Chg, Cng, or Orn; X 12 is W, Y, A, or F; and X 13is G, GABA, or null.

[0059] Alternatively or additionally, the peptide may have the formula R-X1-X2-X3-X4-X5-X6-X7-X8-X9-X 10 -X 11 -X 12 -X 13 -X 14 -X 15 -X 16 In some embodiments, R is NH2, acetylated, stearic acid, palmitic acid, myristic acid, lauric acid, C1-C8 hydrocarbon, C1-C8 fatty acid, or null. In some embodiments, X1 is M, G, beta alanine, norleucine, norvaline, or null. In some embodiments, X2 is W, N-methyl W, R, Y, F, citrulline, or K. In some embodiments, X3 is P, W, N-methyl-W, N-ethyl-W, N-methyl A, N-ethyl A, L, Pip, Aib, Y, or F. In some embodiments, X4 is E, Q, N, or D. In some embodiments, X5 is S, alpha methyl, S, K, D, Orn, T, or E. In some embodiments, X6 is I, Chg, H, or L. In some embodiments, X7 is L or I. In some embodiments, X8 is D, N, E, or Q. In some embodiments, X9 is D, E, K, Q, or Orn. 10 is H or methyl-H. In some embodiments, X 11 is V, alpha methyl V, Chg, L, I, or norvaline. 12 is Q, Aib, S, R, or N. In some embodiments, X 13 is R, K, citrulline, Orn, D, or E. In some embodiments, X 14 is V, I, L, or norvaline. 15 is W, Y, or F. In some embodiments, X 16is R, G, or null. In some embodiments, R is NH2, acetylated, stearic acid, palmitic acid, myristic acid, lauric acid, C1-C8 hydrocarbon, C1-C8 fatty acid, or null; X1 is M, G, beta alanine, norleucine, norvaline, or null; X2 is W, N-methyl W, R, Y, F, citrulline, or K; X3 is P, W, N-methyl-W, N-ethyl-W, N-methyl A, N-ethyl A, L, Pip, Aib, Y, or F; X4 is E, Q, N, or D; X5 is S, alpha methyl S, K, D, Orn, T, or E; X6 is I, Chg, H, or L; X7 is L or I; X8 is D, N, E, or Q; X9 is D, E, K, Q, or Orn; 10 is H or methyl-H; X 11 is V, α-methyl V, Chg, L, I, or norvaline; X 12 is Q, Aib, S, R, or N; X 13 is R, K, Citrulline, Orn, D, or E; X 14 is V, I, L, or norvaline; X 15 is W, Y, or F; and X 16 is R, G, or null.

[0060] In some cases, polar or charged groups can be removed from the peptide to increase affinity or permeability, for example, leucine or isoleucine may often be replaced with cyclohexylglycine.

[0061] In some embodiments, the peptides can be 3 to 80 amino acids in length. In some embodiments, the peptides can be at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, at least 21, at least 22, at least 23, at least 24, at least 25, at least 26, at least 27, at least 28, at least 29, at least 30, at least 31, at least 32, at least 33, at least 34, at least 35, at least 36, at least 37, at least 38, at least 39, at least 40, at least 41, at least 42 , at least 43, at least 44, at least 45, at least 46, at least 47, at least 48, at least 49, at least 50, at least 51, at least 52, at least 53, at least 54, at least 55, at least 56, at least 57, at least 58, at least 59, at least 60, at least 61, at least 62, at least 63, at least 64, at least 65, at least 66, at least 67, at least 68, at least 69, at least 70, at least 71, at least 72, at least 73, at least 74, at least 75, at least 76, at least 77, at least 78, at least 79, at least 80, or at least 81 amino acid residues in length.

[0062] In some embodiments, the peptide is less than 4, less than 5, less than 6, less than 7, less than 8, less than 9, less than 10, less than 11, less than 12, less than 13, less than 14, less than 15, less than 16, less than 17, less than 18, less than 19, less than 20, less than 21, less than 22, less than 23, less than 24, less than 25, less than 26, less than 27, less than 28, less than 29, less than 30, less than 31, less than 32, less than 33, less than 34, less than 35, less than 36, less than 37, less than 38, less than 39, less than 40, less than 41, less than 42, less than 43 less than, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, or 81 amino acid residues in length.

[0063] In some embodiments, the peptide may exhibit homology to TCF4. In some embodiments, the peptide exhibits about 10% to about 100% homology to TCF4. In some embodiments, the peptide exhibits about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, or about 100% homology to TCF4.

[0064] In some embodiments, the peptide may exhibit homology to axin. In some embodiments, the peptide exhibits about 10% to about 100% homology to axin. In some embodiments, the peptide exhibits about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, or about 100% homology to axin.

[0065] Peptide synthesis methods The peptides described herein can be produced by synthetic methods.

[0066] For example, solid phase peptide synthesis (SPPS) allows the rapid assembly of peptide chains by sequential reactions of amino acid derivatives on an insoluble porous support.

[0067] The solid support often comprises small polymeric resin beads functionalized with reactive groups (such as amine or hydroxyl groups) that will be linked to the nascent peptide chain. Because the peptide remains covalently attached to the support throughout the synthesis, excess reagents and by-products can be removed by washing and filtration.

[0068] Each amino acid attached to the N-terminus of the peptide chain can be protected on its N-terminus and side chain using a suitable protecting group such as Boc (acid-labile) or Fmoc (base-labile), depending on the side chain and the protection strategy used (see below).

[0069] A typical SPPS procedure is usually one of repeated cycles of N-terminal group deprotection and coupling reactions. The resin may be washed between each step. First, an amino acid is often coupled to the resin. Next, the amine is usually deprotected and then coupled with the free acid of the second amino acid. This cycle can be repeated until the desired sequence is synthesized. The SPPS cycle may further include a capping step in which the termini of unreacted amino acids may be blocked from reacting. At the end of the synthesis, the crude peptide may be cleaved from the solid support. This step may often involve removing all protecting groups at once using a strong acid or nucleophile such as trifluoroacetic acid. To remove organic soluble by-products, the crude peptide may be precipitated from a non-polar solvent such as diethyl ether. The crude peptide may be purified using reversed-phase HPLC. By-products may be removed using sequential chromatography, such as the MCSGP process, to maximize yields without sacrificing purity levels.

[0070] b. Methods for Cyclizing Peptides The peptides described herein may be cyclic peptides. Cyclic peptides are often stable peptide analogs with strong conformational and in vivo stability. Cyclic peptides may provide some advantages in some situations. For example, cyclic peptides are often more resistant to proteases than their non-cyclic counterparts, and therefore metabolize slower; on the other hand, cyclic peptides have a longer-acting depot effect than their corresponding linear counterparts. Cyclic peptides can be used to mimic the structure of active peptides (e.g., peptide hormones) and can bind to in vivo drug targets.

[0071] 1) Disulfide bridge Cys-Cys thiol oxidation The peptides described herein can be cyclized using disulfide bridges. Peptide disulfide bridges can link two thiol (SH) groups from the side chains of cysteine ​​or cysteine ​​analogs. Unwanted bonds can be prevented by using appropriate protective group chemistry, and specific oxidation can be achieved either intramolecularly or intermolecularly. In general, disulfide bridges can be formed intermolecularly (two peptide molecules are linked via a disulfide bridge), resulting in either homodimers (two identical peptides) or heterodimers (two different peptides), or intramolecularly (cyclization within one peptide molecule).

[0072] 2) Amide bond formation, lactam formation Cyclic peptides can also be synthesized by linking the amino (N) terminus of a peptide to the carboxyl (C) terminus via an amide bond. The amino side chains of Lys and Orn, and the carboxyl side chains of Asp and Glu may also be used to construct cyclic peptides via amide bonds. Depending on the functional groups of the peptide, cyclic peptide synthesis often uses one of four different methods: head-to-tail between the N-terminus and the C-terminus; head-to-side chain between the N-terminus and an internal COOH (e.g., the s-COOH group of Asp or the γ-COOH group of Glu); side chain-to-tail between the internal NH2s and the C-terminus (e.g., the ε-NH2-group of Ly with either the s-COOH group of Asp or the γ-COOH group of Glu).

[0073] 3) Stapled peptide synthesis The peptides described herein can be cyclized using stapled peptide synthesis. Peptide stapling is typically accomplished by incorporating α,α disubstituted unnatural amino acids with terminal olefin tethers of various lengths. Subsequent olefin metathesis creates carbon-carbon bond tethers between amino acid side chains to cyclize the peptide. Alternatively, stapled peptides can be generated using Fmoc solid-phase synthesis chemistry, which is commonly known in the art. Stapled peptides have a chemically locked conformational structure that can mimic the molecular structure typically found at the interface of protein-protein interactions. Once locked into this stable configuration, constraint peptides can penetrate cells and exert their effects on protein targets within the cell. The large surface area of ​​peptides makes them superior to small molecules in their ability to disrupt specific signaling pathways by inhibiting targeted protein-protein interactions.

[0074] 4) Click chemistry for peptide cyclization The peptides described herein can be further cyclized using click chemistry. Using various combinations of protected amino acids modified with alkyne groups, clickable functional groups can be incorporated into peptides during synthesis, followed by click reaction with azide acid. The resulting peptide is separated from the resin to yield a triazole-containing peptide. These functional groups can be further introduced by post-synthetic modification to produce conformationally constrained peptides.

[0075] c. Methods for improving peptide stability The peptides described herein may be modified to improve stability. It has been generally shown that N-methyl modifications can improve the function and stability of peptide ligands (Fiacco et al., Chembiochem 2008, 9(14):2200; Fiacco et al., Chembiochem 2016, 17(17):1643, each of which is incorporated herein by reference for such disclosure). In some embodiments described herein, the peptides are modified with one or more N-methyl analogs to natural amino acids. Incorporation of one or more N-methyl amino acids can increase proteolytic resistance by 10-fold to 10,000-fold over the resistance of peptides composed of natural residues. In some embodiments, the proteolytic resistance of peptides containing one or more N-methyl amino acids is at least 10 times, at least 20 times, at least 30 times, at least 40 times, at least 50 times, at least 60 times, at least 70 times, at least 80 times, at least 90 times, at least 100 times, at least 200 times, at least 400 times, at least 500 times, at least 600 times, at least 700 times, at least 800 times, at least 900 times, at least 1000 times, at least 2000 times, at least 3000 times, at least 4000 times, at least 5000 times, at least 6000 times, at least 7000 times, at least 8000 times, at least 9000 times, at least 10,000 times greater than the resistance of peptides consisting of natural residues.

[0076] III. Characterization Assays A. Surface Plasmon Resonance Surface plasmon resonance (SPR) is an optical phenomenon that allows the detection of unlabeled reactants in real time. SPR-based biosensors such as the Biacore® T100 system (Biacore®, Uppsala, Sweden) can be used to determine active concentrations and characterize molecular interactions in terms of both affinity and chemical kinetics. The maximum binding capacity of a surface (response at saturation) can be calculated using the Biacore® system (GE) instructions. Binding levels are determined over a range of analyte concentrations from at least 20% to 80% saturation of the surface. On rates, off rates, and KD are calculated using Biacore® Insight Evaluation So0ftware (GE). The concentration at 50% saturation is calculated from these dilution series and is denoted as KD50. Binding kinetics and thermodynamics of peptides selected from the libraries described herein are displayed in Table 5. Exemplary methods are described in Example 11.

[0077] In some examples, the peptides described herein comprise a binding affinity (KD) of greater than about 30 uM. In some examples, the peptides described herein comprise a binding affinity (KD) of less than about 30 uM, less than about 10 uM, less than about 5 uM, less than about 1 uM, less than about 500 nm, less than about 400 nM, less than about 300 nm, less than about 200 nm, less than about 150 nm, less than about 100 nm, less than about 50 nm, or less than about 10 nm.

[0078] B. Fluorescence Polarization (FP) Fluorescence polarization (FP) is another method used to analyze protein-ligand interactions. This method measures the change in fluorescence light emitted by a labeled compound that is not bound to another compound in solution versus that that is bound to the other compound. Binding of a fluorescent molecule to the complex increases polarization due to restricted rotation of the fluorescent tracer. Competitive FP is often utilized in drug screens because it is formatted for high-throughput screens in plate format. This immunological assay is performed by introducing a non-bound drug that competes for binding to the labeled competitive ligand complex, resulting in a decrease in polarization. TCF4 protein is utilized in the competitive assay because (1) this protein affects full activation of the Wnt pathway and (2) shares homology with β-catenin inhibitors. An exemplary method is described in Example 13. Data was analyzed and plotted in Microsoft Excel (Microsoft, Redmond, WA). The resulting KD50 was calculated as shown in Table 5.

[0079] C. Wnt-dependent luciferase reporter assay Luciferase reporter assay is commonly used as a tool to test gene expression at the transcription level. Peptides from the library described herein were characterized for their ability to suppress transcription in cells with luciferase expression controlled by a Wnt-responsive promoter. Peptides were added to cells transfected with a luciferase gene downstream of a Wnt-responsive promoter. Cells were stimulated with Wnt3a protein and incubated with peptides from the library described herein. Luminescence was recorded to determine the peptide with the highest β-catenin inhibitory activity. EC50 values ​​were calculated based on the inhibition of luminescence and are expressed as concentration (uM) in Table 5.

[0080] D. Cell proliferation assay Peptides that inhibit β-catenin cause reduced cell viability in a colon cancer cell line (SW480) and endometriotic lesions excised from animal models of the disease. Peptides from the libraries described herein are shown in Table 5 to have IC50 values ​​measured using a luminescent cell viability assay (Cell Titer-Glo®, Promega, Madison, WI). 50 The effects on cell viability, expressed as percentile, were characterized in terms of their effect on cell viability.

[0081] In some examples, the peptides described herein have an IC of greater than 30 uM. 50 In some examples, the peptides described herein have an IC of less than about 30 uM, less than about 25 uM, less than about 22.5 uM, less than about 20 uM, less than about 15 uM, less than about 10 uM, less than about 7.5 uM, less than about 5 uM, or less than about 1 uM. 50 Includes.

[0082] IV. Administration A. Methods of Administration and Treatment Regimens In one embodiment, the inhibitor compositions described herein are used in the preparation of a medicament for treating a disease, disorder, or condition in a mammal that would benefit from administration of a β-catenin inhibitor. A method of treating any of the diseases or disorders described herein in a mammal in need of such treatment comprises administering to said mammal a pharmaceutical composition comprising a therapeutically effective amount of at least one inhibitor described herein.

[0083] Diseases, disorders, or conditions that may benefit from treatment with the compositions described herein include, but are not limited to, endometriosis, endometrial lesions, endometriomas, superficial endometriotic implants, deep endometriosis, chronic pain, central sensitization, myofascial pain, adnexal masses, infertility, dysmenorrhea, genetic predisposition, non-menstrual pelvic-abdominal pain, dyspareunia, bowel symptoms (diarrhea, cramps, constipation), dysentery (difficulty passing stool), ovarian masses or tumors, painful bladder symptoms, and dysuria.

[0084] In some embodiments, compositions comprising the inhibitors described herein are administered for preventive and / or therapeutic treatment. In some therapeutic applications, the compositions are administered to a patient already suffering from a disease or condition in an amount sufficient to cure or at least partially prevent at least one of the symptoms of the disease or condition. The amount effective for this use depends on the severity and course of the disease or condition, previous treatments, the patient's health status, weight, and response to the drug, and the judgment of the treating physician. Therapeutically effective amounts are optionally determined by methods including, but not limited to, dose escalation and / or dose ranging clinical trials.

[0085] In prophylactic applications, compositions containing the inhibitors described herein are administered to patients susceptible to or at risk of a particular disease, disorder, or condition. Such an amount is defined as a "prophylactically effective amount or dosage." In this application, the exact amount will vary depending on the patient's health, weight, and the like. When used in patients, the effective amount for this use will depend on the severity and course of the disease, disorder, or condition, previous treatments, the patient's health and response to the drugs, and the judgment of the treating physician. In one aspect, prophylactic treatment involves administering a pharmaceutical composition containing the inhibitors described herein to a mammal that has previously experienced at least one symptom of the disease being treated and is now in remission, to prevent the recurrence of the disease or condition.

[0086] In certain embodiments where the patient's condition is not improved, the inhibitor is administered chronically, i.e., for an extended period of time, including the patient's life, at the physician's discretion, to ameliorate or otherwise manage or limit the symptoms of the patient's disease or disorder.

[0087] In certain embodiments where the patient's condition has improved, the dosage of the administered drug is temporarily reduced or temporarily stopped for a period of time (i.e., a "drug holiday"). In certain embodiments, the length of the drug holiday is between 2 days and 1 year, including, by way of example only, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 10 days, 12 days, 15 days, 20 days, 28 days, or longer than 28 days. Dose reductions during drug holidays can be, by way of example only, 10%-100%, including, by way of example only, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, and 100%.

[0088] Once the patient's condition has improved, a maintenance dose is administered as needed. Thereafter, in certain embodiments, the dosage or frequency of administration, or both, depending on the symptoms, is reduced to a level at which the improved disease, disorder, or condition is maintained. In certain embodiments, however, patients require intermittent treatment on a long-term basis upon recurrence of symptoms.

[0089] The amount of a given drug that corresponds to such an amount will vary depending on factors such as the particular inhibitor, the disease state and its severity, the identity of the subject or host requiring treatment (e.g., weight, sex), etc., but will nevertheless be determined by the particular circumstances surrounding the case, including, for example, the particular drug being administered, the route of administration, the disease being treated, and the subject or host being treated.

[0090] In general, dosages used for adult human treatment typically range from about 0.01 mg to about 5000 mg per day. In some aspects, dosages utilized for adult human treatment are from about 0.01 mg to about 1000 mg per day. In some embodiments, the desired dosage is provided in a single dose or in divided doses administered simultaneously or at appropriate intervals, for example, as two, three, four or more subdoses per day. In other embodiments, the compositions described herein are formulated for sustained release over a period of hours, days, or months. In some embodiments, the composition is formulated for delivery over 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, 24 hours, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, 1 year, 2 years, 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, 11 years, or 12 years.

[0091] In some embodiments, a suitable daily dosage for the inhibitors described herein is about 0.01 to 50 mg / kg of body weight. In some embodiments, the daily dosage or amount of active ingredient in the dosage form will be lower or higher than the ranges set forth herein, based on many variables related to the individual treatment regimen. In various embodiments, the daily dosage and unit dosage will vary depending on many variables, including, but not limited to, the activity of the inhibitor used, the disease or condition being treated, the mode of administration, the requirements of the individual subject, the severity of the disease or condition being treated, and the judgment of the physician.

[0092] Toxicity and therapeutic efficacy of such treatment regimens are determined by standard pharmaceutical procedures in cell cultures or experimental animals, including, but not limited to, determining LD50 and ED50. The dose ratio between toxic and therapeutic effects is the therapeutic index, which is expressed as the ratio between LD50 and ED50. In some embodiments, data obtained from cell culture assays and animal studies are used to formulate a therapeutically effective daily dosage range and / or therapeutically effective unit dose for use in mammals, including humans. In some embodiments, the daily dosage of the inhibitors described herein is within a range of blood concentrations that includes the ED50 of minimal toxicity. In some embodiments, the daily dosage range and / or unit dose varies within this range depending on the dosage form used and the route of administration utilized.

[0093] In any of the foregoing aspects, there are further embodiments in which an effective amount of an inhibitor described herein is (a) administered systemically to a mammal, and / or (b) administered orally to a mammal, and / or (c) administered intravenously to a mammal, and / or (d) administered by injection to a mammal, and / or (e) administered locally to a mammal, and / or (f) administered non-systemically or locally to a mammal.

[0094] In any of the foregoing aspects, there are further embodiments that include a single administration of an effective amount of the compound, including further embodiments in which (i) the inhibitor is administered once daily; or (ii) the inhibitor is administered multiple times to the mammal over the course of a day. In further aspects, there are embodiments in which the inhibitor is administered continuously over a period of time.

[0095] In any of the foregoing aspects, there are further embodiments that include multiple administrations of an effective amount of the inhibitor, including further embodiments in which (i) the inhibitor is administered continuously or intermittently, such as in a single dose; (ii) the interval between multiple doses is every 6 hours, (iii) the inhibitor is administered to the mammal every 8 hours; (iv) the inhibitor is administered to the mammal every 12 hours; or (v) the inhibitor is administered to the mammal every 24 hours.

[0096] In any of the above-mentioned aspects, there are further embodiments that include the inhibitor incorporated in the delivery vehicle to provide sustained delivery of the inhibitor.In some embodiments, delivery is sustained for about 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, 24 hours, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, 1 year, 2 years, 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, 11 years, or 12 years. In some embodiments, the inhibitor is incorporated into a transdermal patch, tape, suppository, vaginal pessary, vaginal tampon, vaginal ring, vaginal strip, vaginal capsule, vaginal tablet, vaginal pessary, vaginal cup, vaginal sponge, or intrauterine device.

[0097] In further or alternative embodiments, the method includes a drug holiday, during which administration of the inhibitor is temporarily suspended or the amount of inhibitor being administered is temporarily reduced; at the end of the drug holiday, administration of the inhibitor is resumed. In one embodiment, the length of the drug holiday varies from 2 days to 1 year.

[0098] In some instances, it is appropriate to administer at least one inhibitor described herein in combination with one or more therapeutic agents.Exemplary additional therapeutic agents may include hormone therapeutic agents, including but not limited to birth control, combination birth control, selective progesterone receptor antagonist, selective progesterone receptor agonist, gonadotropin releasing hormone receptor antagonist, gonadotropin releasing hormone receptor agonist, antiretroviral drugs, anti-neoplastic drugs, anti-inflammatory drugs, nonsteroidal anti-inflammatory drugs, or any combination thereof.

[0099] In one embodiment, the therapeutic effectiveness of one of the inhibitors described herein is enhanced by administration of an adjuvant (i.e., the adjuvant has minimal therapeutic effect by itself, but when combined with another therapeutic agent, enhances the overall therapeutic effect on the patient), or in some embodiments, the effect experienced by the patient is increased by administering one of the inhibitors described herein with another agent (which also includes a treatment regimen) that also has a therapeutic effect.

[0100] In one particular embodiment, the inhibitor described herein is co-administered with a second therapeutic agent.In some cases, the inhibitor described herein and the second therapeutic agent can modulate different aspects of the disease, disorder, or illness being treated, thereby resulting in a greater overall effect than administering either therapeutic agent alone.In any case, regardless of the disease, disorder, or illness being treated, the overall effect that the patient experiences can be additive of the two therapeutic agents, or the patient can experience a synergistic effect.

[0101] In certain embodiments, various therapeutically effective amounts of the inhibitors disclosed herein are utilized in formulating pharmaceutical compositions and / or in treatment regimens in which the inhibitors disclosed herein are administered in combination with one or more additional agents, such as additional therapeutically effective drugs, adjuvants, etc. The therapeutically effective amounts of drugs and other agents used in combination therapy regimens are optionally determined by means similar to those specified above for the active ingredients themselves. Additionally, the methods of prevention / treatment described herein encompass the use of metronomic dosing, i.e., providing more frequent, lower doses to minimize toxic side effects. In some embodiments, combination treatment regimens include treatment regimens in which administration of the inhibitors described herein is initiated before, during, or after treatment with a second agent described herein, and continues until any time during or after the end of treatment with the second agent. It also includes treatments in which the inhibitors described herein and the second agent used in combination are administered simultaneously or at different times and / or at decreasing or increasing intervals during the treatment period. Concomitant treatments also include scheduled treatments that are started and stopped at various times to aid in the clinical management of the patient.

[0102] It will be understood that the dosage regimen to treat, prevent, or ameliorate the condition for which relief is sought may be modified to accommodate a variety of factors (e.g., the disease, disorder, or illness from which the subject suffers; the age, weight, sex, diet, and medical condition of the subject). Thus, in some instances, the dosage regimen actually utilized will vary and, in some embodiments, will deviate from the dosage regimens set forth herein.

[0103] With respect to the combination therapies described herein, the dosage of the co-administered inhibitor will vary depending on the particular co-medication used, the particular drug used, the disease or condition being treated, etc. In additional embodiments, when co-administered with one or more other therapeutic agents, the inhibitors provided herein are administered simultaneously or sequentially with the one or more other therapeutic agents.

[0104] In combination therapy, the multiple therapeutic agents (one of which is one of the inhibitors described herein) are administered in any order or simultaneously. When administration is simultaneous, the multiple therapeutic agents may be provided in a single, unified form or in multiple forms (e.g., as a single pill, or as two separate pills), by way of example only.

[0105] In conjunction with the combination therapy, the inhibitors described herein are administered before, during, or after the onset of a disease or condition, and the timing of administering the pharmaceutical composition containing the inhibitor can vary. Thus, in one embodiment, the inhibitors described herein are used as prophylactics and are administered continuously to subjects prone to developing a disease or condition to prevent the onset of the disease or condition. In another embodiment, the inhibitors and compositions are administered to the subject during or as soon as possible after the onset of symptoms. In certain embodiments, the inhibitors described herein are administered as soon as practicable after the onset of a disease or condition is detected or suspected, and for the period of time required to treat the condition. In some embodiments, the period of time required for treatment can vary, and the treatment period is adjusted to the specific needs of each subject. For example, in certain embodiments, the inhibitors described herein or formulations containing the inhibitors are administered for at least 2 weeks, about 1 month to about 5 years.

[0106] B. Pharmaceutical Compositions In some embodiments, the inhibitors described herein are formulated into pharmaceutical compositions. Pharmaceutical compositions are formulated in a conventional manner using one or more pharma- ceutical acceptable inactive ingredients that facilitate the processing of active inhibitors into pharmaceutical preparations. Appropriate formulations depend on the route of administration selected. Summary of pharmaceutical compositions described herein can be found, for example, in: The Science and Practice of Pharmacy, Nineteenth Ed (Easton, Pa.: Mack Publishing Company, 1995); Hoover, John E., Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pennsylvania 1975; Liberman, HA and Lachman, L., Eds., Pharmaceutical Dosage Forms, Marcel Decker, New York, NY, 1980; and Pharmaceutical Dosage Forms and Drug Delivery Systems, Seventh Ed. (Lippincott Williams & Wilkins 1999), which are incorporated herein by reference for such disclosure.

[0107] In some embodiments, the inhibitors described herein are administered alone or in pharmaceutical compositions in combination with pharma- ceutically acceptable carriers, excipients, or diluents. Administration of the inhibitors and compositions described herein can be achieved by a method that allows the inhibitor to be delivered to the site of action. Such methods include, but are not limited to, delivery via enteral routes (including oral, gastric, or duodenal feeding tubes, rectal suppositories, and rectal enemas), parenteral routes (including intraarterial, intracardiac, intradermal, intraduodenal, intramedullary, intramuscular, intraosseous, intraperitoneal, intrathecal, intravascular, intravenous, intravitreal, epidural, and subcutaneous injection or infusion), inhalation, transdermal, transmucosal, sublingual, buccal, and topical (including epithelial, dermal, enema, eye drops, ear drops, intranasal, vaginal, and intrauterine) administration, although the most suitable route may depend, for example, on the disease and disorder of the recipient. By way of example only, the inhibitors described herein may be administered locally to the area in need of treatment, for example, by local infusion during surgery, topical application such as a cream or ointment, injection, catheter, implant, or insertion device. Administration may also be by direct injection at the site of the diseased tissue or organ.

[0108] In some embodiments, pharmaceutical compositions suitable for oral administration are presented as discrete units such as capsules, cachets, or tablets, each containing a predetermined amount of the active ingredient; as a powder or granules; as a solution or suspension in an aqueous or non-aqueous liquid; or as an oil-in-water emulsion or a water-in-oil emulsion in an oil liquid. In some embodiments, the active ingredient is provided as a bolus, electuary, or paste.

[0109] Pharmaceutical compositions that can be used orally include tablets, push-fit capsules made of gelatin, and sealed soft capsules made of gelatin and a plasticizer, such as glycerol or sorbitol. Tablets may be made by compression or molding, optionally with one or more accessory ingredients. Compressed tablets can be prepared by compressing in a suitable machine the active ingredient in a free-flowing form such as powder or granules, optionally mixed with a binder, inert diluent, or lubricant, surface active agent, or dispersing agent. Molded tablets can be produced by molding in a suitable machine a mixture of powdered inhibitor moistened with an inert liquid diluent. In some embodiments, the tablets are coated or scored and formulated to provide a delayed or controlled release of the active ingredient therein. All formulations for oral administration may be in dosages suitable for such administration. Push-fit capsules can contain the active ingredient in combination with a filler such as lactose, a binder such as starch, and / or a lubricant such as talc or magnesium stearate, and optionally stabilizers. In soft capsules, active inhibitors can be dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin, or liquid polyethylene glycol. In some embodiments, stabilizers are added. Dragee cores are provided with suitable coatings. For this purpose, concentrated sugar solutions can be used, which can optionally contain gum arabic, talc, polyvinylpyrrolidone, carbopol gel, polyethylene glycol, and / or titanium dioxide, lacquer solutions, and suitable organic solvents or solvent mixtures. Dyes or pigments can be added to tablets or dragee coatings for identification or to characterize different combinations of active inhibitor doses.

[0110] In some embodiments, the pharmaceutical composition is formulated for parenteral administration by injection, for example, bolus injection or continuous infusion. The preparation for injection may be provided in unit dosage form, for example, in ampoules or multi-dose containers, together with additional preservatives. The composition may take the form of a suspension, solution, or emulsion in an oily or aqueous vehicle, and may contain formulating agents such as suspending, stabilizing, and / or dispersing agents. The composition may be provided in unit or multi-dose containers, for example, sealed ampoules and vials, and may be stored in powder form or in a freeze-dried (lyophilized) condition requiring only the addition of a sterile liquid carrier, for example, saline or pyrogen-free distilled water, immediately prior to use. Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules, and tablets of the kind described above.

[0111] Pharmaceutical compositions for parenteral administration include aqueous and non-aqueous (oily) sterile injection solutions of active inhibitors, which may contain antioxidants, buffers, bacteriostats, and solutes that render the formulation isotonic with the blood of the desired recipient; aqueous and non-aqueous sterile suspensions, which may contain suspending agents and thickening agents. Suitable lipophilic solvents or vehicles include fatty oils such as sesame oil, synthetic fatty acid esters such as ethyl oleate or triglycerides, or liposomes. Aqueous injection suspensions may contain substances that increase the viscosity of the suspension, such as sodium carboxymethylcellulose, sorbitol, or dextran. Optionally, the suspension may also contain suitable stabilizers or agents that increase the solubility of the inhibitor to allow for the preparation of highly concentrated solutions.

[0112] The pharmaceutical composition can also be formulated as a depot preparation. Such long-acting preparations can be administered by implantation (e.g., subcutaneously or intramuscularly) or intramuscular injection. Thus, for example, the inhibitor can be formulated with suitable polymeric or hydrophobic materials (e.g., as an emulsion in an acceptable oil) or ion exchange resins, or as sparingly soluble derivatives, e.g., as sparingly soluble salts.

[0113] For buccal or sublingual administration, the compositions may take the form of tablets, lozenges, pastilles, or gels formulated in a conventional manner. Such compositions may contain the active ingredient in a flavored base, such as sucrose and acacia or tragacanth.

[0114] The pharmaceutical compositions may also be formulated in rectal or vaginal compositions, such as suppositories or retention enemas, for example, by including conventional suppository bases, such as cocoa butter, polyethylene glycol, or other glycerides. Upon insertion, the suppository base liquefies or becomes water-miscible at body temperature so that the ingredients can be in contact with the mucosa for a period of time long enough to have a therapeutic effect. The weight percentage of the suppository base depends on the size of the opening of the human and / or animal body, the dosage composition required to have a therapeutic effect, and its physicochemical properties that allow it to remain solid at room temperature or below. In some embodiments, the suppository includes more than about 50% to 99% by weight, or more than about 75% to 99% by weight, of the suppository base. In some embodiments, the suppository includes about 75% to about 98% by weight of polyethylene glycol. In some embodiments, the suppository includes about 2% to about 25% by weight of polysorbate. Suppository bases have a molecular weight ranging from about 400 to about 5000, or from about 950 to about 3700 (US 2009 / 0311290, which is incorporated herein by reference for such disclosure).

[0115] In some cases, the pharmaceutical composition includes an absorption enhancer, such as sodium caprate. Some such cases include compositions and dosage forms for rectal or vaginal use, as described herein.

[0116] The pharmaceutical compositions can be administered topically, rectally, i.e., by non-systemic administration. This includes applying the inhibitors of the present disclosure to the external epidermis or buccal cavity, and instilling such inhibitors into the rectum or vagina so that the inhibitors do not enter the bloodstream as much as possible. In contrast, systemic administration refers to oral, intravenous, intraperitoneal, and intramuscular administration.

[0117] Pharmaceutical compositions suitable for topical administration include liquid or semi-liquid formulations suitable for penetration through the skin, e.g., solutions, lotions, coagulants, creams, ointments, gels, foams, transdermal patches, powders, solids, sponges, tapes, vapors, pastes, tinctures, microparticles, microcapsules, nanoparticles, liposomes, or emulsions, including those suitable for vaginal or rectal delivery. The active ingredient may comprise from 0.001% w / w to 10% w / w of the formulation, for example, 1% to 2% by weight, for topical administration.

[0118] Pharmaceutical compositions for administration by inhalation are conveniently delivered from an insufflator, nebulizer, pressurized pack, or other convenient means of delivering an aerosol spray. Pressurized packs can contain a suitable propellant, such as dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide, or other suitable gas. In the case of a pressurized aerosol, the dosage unit can be determined by providing a valve to deliver a measured amount. Alternatively, for administration by inhalation or insufflation, the formulation can take the form of a dry powder composition, for example, a powder mix of the inhibitor and a suitable powder base, such as lactose or starch. The powder composition can be presented in unit dosage form, for example, capsules, cartridges, gelatin, or blister packs, from which the powder can be administered using an inhaler or insufflator.

[0119] In some embodiments, the inhibitors disclosed herein are formulated to achieve delivery to specific regions of the gastrointestinal tract.For example, the inhibitors disclosed herein are formulated for oral delivery by bioadhesive polymers, pH-sensitive coatings, time-dependent biodegradable polymers, microflora activation systems, etc., to achieve delivery of the inhibitors to specific regions of the gastrointestinal tract.

[0120] In some embodiments, the inhibitors disclosed herein are formulated to achieve delivery to specific regions of the urogenital or anorectal mucosa. For example, the inhibitors disclosed herein are formulated for intravaginal delivery with bioadhesive polymers, pH-sensitive coatings, time-dependent biodegradable polymers, microflora activation systems, etc., to achieve delivery of the inhibitor to specific regions of the urogenital system. In some embodiments, the inhibitors disclosed herein are formulated to provide controlled release of the inhibitor. Controlled release refers to the release of the inhibitor from a dosage form in which the inhibitor is incorporated as described herein according to a desired profile over an extended period of time. Controlled release characteristics include, for example, sustained release, extended release, pulsatile release, and delayed release characteristics. In contrast to immediate release compositions, controlled release compositions allow delivery of the drug to a subject over an extended period of time according to a predefined profile. Such release rates provide therapeutically effective levels of the drug for an extended period of time, thereby providing a longer duration of pharmacological response, while minimizing side effects compared to conventional rapid release dosage forms. This long-term response offers many inherent benefits that are not achieved with the corresponding short-acting immediate release formulations.

[0121] Approaches to deliver intact therapeutic inhibitors to specific regions of the urogenital system (such as the vagina) or gastrointestinal tract (such as the colon) include the following: (i) Polymer coating: Intact molecules can be delivered to the colon without being absorbed in the upper part of the intestine by coating the drug molecule with a suitable polymer that degrades only in the colon. In addition, polymer coatings can provide a protective or controlled release profile for vaginal formulations. (ii) Coating with pH-sensitive polymers: Intestinal, colonic and vaginal targeted delivery systems are based on coating tablets or pellets which are filled into conventional hard gelatin capsules. The most commonly used pH-dependent coating polymers are methacrylic acid copolymers (commonly known as Eudragit® S), more specifically Eudragit® L and Eudragit® S. Eudragit® L100 and S100 are copolymers of methacrylic acid and methyl methacrylate. (iii) coating with biodegradable polymers; (iv) embedding in a matrix; (v) embedding in biodegradable matrices and hydrogels; (vi) embedding in a pH-sensitive matrix; (vii) timed release systems; (viii) Redox-sensitive polymer; (ix) bioadhesive systems; (x) particulate coating; (xi) Osmotically controlled drug delivery.

[0122] Another approach to vaginal and colonic targeted drug delivery or controlled release systems involves embedding drugs in polymeric matrices to entrap and release the drug in the colon. These matrices can be pH-sensitive or biodegradable. Matrix-based systems such as multi-matrix (MMX)-based delayed release tablets ensure drug release in the vagina or colon.

[0123] C. Dosage form The compositions and methods described herein may include delivering therapeutic agents by various dosage forms and devices, including those described below. Many of the dosage forms described below have several advantages. These may include local delivery of therapeutic inhibitors. Local delivery can reduce side effects that may be associated with systemic delivery and off-target effects. In some cases, the dosage forms and devices described herein can improve patient compliance, for example, by providing long-term or continuous delivery of the inhibitors described herein. In some cases, these dosage forms and devices can further increase tolerability and safety.

[0124] 1. Intravaginal ring The peptides and compositions described herein may be incorporated into an intravaginal ring for delivery. Delivery of therapeutic agents via an intravaginal ring (IVR) allows for localized delivery and improves the safety and tolerability of therapeutic agents. IVR may further have multiple advantages: bypass gastrointestinal absorption and liver / renal first pass, lower effective dose, continuous delivery and / or controlled release profile, extended time between doses, reduced side effects, low serum drug concentrations, patient self-administration, and improved patient satisfaction. IVR can successfully deliver drugs, including hydrophilic and macromolecular agents, to organs affected by EMS, including deep EMS (DIE). The biodistribution of the intravaginal ring delivery system is by tissue absorption rather than serum, and can reach all organs affected by EMS in the peritoneal cavity.

[0125] Vaginal rings usually consist of an inert elastomeric ring coated with another layer of elastomer containing the drug to be delivered. The ring can be easily inserted and left in place for a desired period of time, then removed by the user. The ring can be solid or hollow containing a therapeutic component, or it can be a porous material from which the drug is released. The ring can optionally include a third outer rate-controlling elastomeric layer that does not contain a drug. Optionally, the third ring can contain a second drug for dual release rings. Drugs can be incorporated into polyethylene glycol throughout the silicone elastomeric ring to act as a reservoir for the drug to be delivered. In some cases, the IVR can include silicone, compressed tablets, or freeze-dried gels.

[0126] Pessaries, cups, strips, tablets, and suppositories are other examples of drug delivery systems that may be used in the present disclosure. These systems have been used to deliver vaginal contraceptives and have been widely described in the literature.

[0127] Another example of a delivery system is a vaginal sponge and vaginal foam. The desired pharmaceutical agent can be incorporated into a silicone matrix that is coated onto a cylindrical drug-free polyurethane vaginal sponge, as described in the literature.

[0128] In some embodiments, the IVR comprises the peptide or composition described herein formulated as a suppository, solution, lotion, concomitant, cream, ointment, gel, foam, transdermal patch, powder, solid, sponge, tape, paste, tincture, emulsion, microparticle, microcapsule, nanoparticle, liposome, or capsule containing microparticle, microcapsule, nanoparticle, or liposome. The IVR is further modified to include various delivery vehicles such as silicone inserts, compressed tablets, or lyophilized gels to optimize the release profile of hydrophilic or high molecular weight inhibitors, such as peptides, proteins, or antibodies. (Morrow, et.al., Eur J Pharm Biopharm, 2011 Jan; 77(1):3-10, which is incorporated herein by reference for such disclosure). The IVR may also include one or more absorption enhancers, such as sodium caprate.

[0129] In some embodiments, the IVR described herein is formulated to contain about 0.01 mg to about 5000 mg of the inhibitor. In some embodiments, the IVR may contain about 0.01 mg, about 0.05 mg, about 0.1 mg, about 0.5 mg, about 1 mg, about 5 mg, about 10 mg, about 20 mg, about 40 mg, about 60 mg, about 80 mg, about 100 mg, about 150 mg, about 200 mg, about 400 mg, about 600 mg, about 800 mg, about 1000 mg, about 1200 mg, about 1400 mg, about 1600 mg, about 1800 mg, about 2000 mg, about 2500 mg, about 3000 mg, about 3500 mg, about 4000 mg, about 4500 mg, or about 5000 mg of the inhibitor.

[0130] In some embodiments, the IVR described herein is formulated to deliver about 0.01 mg to about 1000 mg of inhibitor per day. In some embodiments, the IVR is formulated to deliver about 0.01 mg, about 0.05 mg, about 0.1 mg, about 0.5 mg, about 1 mg, about 5 mg, about 10 mg, about 20 mg, about 40 mg, about 60 mg, about 80 mg, about 100 mg, about 150 mg, about 200 mg, about 400 mg, about 600 mg, about 800 mg, or about 1000 mg of inhibitor per day.

[0131] 2. Vaginal tampons The peptides and compositions described herein can be incorporated into a tampon device for delivery. The tampon device typically includes a vaginal tampon having a proximal end and a distal end. The cup-shaped porous foam portion at the distal end fits around the cervix and contains the peptides or compositions described herein for delivery. The device may further include a non-absorbent axial tube having a distal opening and extending through the porous foam cup into the tampon to direct blood flow to the absorbent material. Optionally, a retrieval string or tape is also included that is tethered to the tampon device. The absorbent vaginal tampon may contain or be coated with the peptides or compositions described herein and can be used as a medicated tampon for delivery.

[0132] 3. Pessaries / suppositories / compressed tablets The peptides and compositions described herein can be incorporated into a solid for local delivery. The solid can take the form of a pessary or a vaginal or anal suppository. Upon reaching body temperature, the solid dosage form may melt. Alternatively, the solid can retain its structure and release the incorporated compositions described herein. The solid can be a pessary designed to provide support in the vagina. The vaginal sponge can be embedded with the compositions described herein for intravaginal delivery.

[0133] 4. Topical medications The peptides and compositions described herein can be incorporated into topical formulations for delivery. Topical agents are applied to body surfaces such as skin or mucous membranes. In some examples, body surfaces include, but are not limited to, epithelial tissue, mucosal tissue, peritoneum, perimetrium, and endometrium. The inhibitor is absorbed by the body surface to achieve a local or systemic effect. Topical agents are optionally formulated in the following classes: topical solutions; lotions; concomitants; creams; ointments; gels; foams; transdermal patches; powders; solids; sponges; tapes; vapors; pastes; or tinctures. Topical solutions may typically be administered as low-viscosity washes, sprays, or drops with a water or alcohol base. Lotions are thicker and more emollient than solutions. They are usually oils mixed in water and may have less alcohol than solutions. Concussives are mixtures that divide into two or three parts over time. The mixture may be oil mixed with a water-based solution and must be shaken into a suspension before use. Creams are emulsions with approximately equal proportions of oil and water. Ointments are homogeneous, viscous, semi-solid preparations, most commonly thick oils (80% oil-20% water) with high viscosity and greasy properties. Ointments may contain a hydrocarbon base, an absorption base, a water-soluble base, an emulsifying base, or a vegetable oil, or any combination thereof. Ointments are formulated in bases including, but not limited to, a hydrocarbon base such as solid paraffin, soft paraffin, microcrystalline wax, or ceresin; an absorbent base such as wool fat or beeswax; a water-soluble base such as macrogol 200, 300, or 400; an emulsifying base such as emulsifying wax or cetrimide; or a vegetable oil such as olive oil, coconut oil, sesame oil, almond oil, or peanut oil.

[0134] Gels are semi-solid emulsions. Non-limiting examples of useful emulsifiers include acrylic acid polymers (such as Carbomer brand thickeners manufactured by Voveon, Inc., e.g., Carbomer 934P), polyoxyethylene-10-stearyl ether, polyoxyethylene-20-stearyl ether, cetostearyl alcohol, cetyl alcohol, cholesterol, diglycol stearate, glyceryl monostearate, glyceryl stearate, polyglyceryl-3-oleate, hydroxypropyl cellulose, hydroxypropylmethylcellulose, lanolin, polyoxyethylene lauryl ether, methylcellulose, polyoxyethylene stearate, polysorbates, propylene glycol monostearate, sorbitan esters, stearic acid, or mixtures of two or more thereof.

[0135] The amount of emulsifier in the topical formulation may range from about 1 to about 40% by weight, and in some embodiments, from about 5 to about 30% by weight, based on the total weight of the topical formulation.

[0136] The gel formulations described herein may contain one or more gelling agents.Non-limiting examples of useful gelling agents include carboxylic acid polymers, including acrylic acid polymers crosslinked with crosslinking materials such as aryl ether of sucrose (e.g., carbomer brand thickeners), cetostearyl alcohol, hydroxymethylcellulose, polyoxyethylene-polyoxypropylene copolymer, sodium carboxymethylcellulose, polyvinylpyrrolidone, or mixtures of two or more thereof.

[0137] The amount of gelling agent in a topical gel formulation may range from about 0.1 to about 10% by weight, and in some embodiments, from about 0.1 to about 1% by weight, based on the total weight of the topical formulation.

[0138] The gel formulations described herein may further include one or more alkalizing agents, such as sodium hydroxide, in an amount of less than about 2% by weight, as an activator of gelation.

[0139] The formulation may contain one or more additional excipients known in the art, such as water and thickening agents, such as colloidal silicon dioxide.

[0140] Thermoreversible gels are liquid formulations that become a gel when inserted into the rectum or vagina. Thermoreversible gels allow for easier administration and positioning than traditional suppositories or pessaries and can prevent leakage of the dosage form. Thermoreversible gels are formulated as polymer solutions consisting of thermoreversible polymers (e.g., poloxamer combined with a mucoadhesive polymer that allows the gel to adhere to the mucosa). In situ thermoreversible liquid gel formulations, also called thermoreversible "liquid suppositories", are liquid at low temperatures (<10C) and become a gel at body temperature.

[0141] Active agents may be incorporated into surgical tape for occlusive dressings. Drugs may be applied as ointments or gels to reach mucous membranes by evaporation. Pastes combine oils, water, and powders. Tinctures typically contain a high percentage of alcohol for application to the skin.

[0142] 5. Transdermal patches and films The peptides and compositions described herein can be incorporated into transdermal patch delivery, which provides controlled release of drug through a porous membrane covering a reservoir of drug or by dissolving a thin layer of drug embedded in the patch adhesive at body temperature.

[0143] The compositions described herein can be incorporated into film for delivery.Film is a thin and small polymer preparation that can be easily inserted into vaginal cavity without applicator and without causing discomfort.Vaginal film is easier to apply than other types of vaginal preparations such as pessary, foam and gel (Rohan LC et al.,AAPS Journal 2009;11:78-87, which is incorporated herein by reference for such disclosure).

[0144] 6. Intrauterine devices The peptides and compositions described herein can be incorporated into an intrauterine device (IUD), a small, often T-shaped device that is inserted into the uterus for delivery. IUDs typically contain copper, progestogens, or levonorgestrel. The compositions described herein are incorporated into the IUD device and slowly released over time.

[0145] 7. Systemic Delivery (Injection) The peptides and compositions described herein can be formulated for systemic delivery.

[0146] Parenteral injections may be formulated for bolus injection or continuous infusion. Pharmaceutical compositions may be in a form suitable for parenteral injection as sterile suspensions, solutions, or emulsions in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabilizing, and / or dispersing agents. Pharmaceutical formulations for parenteral administration include aqueous solutions of the peptides described herein in water-soluble form. Suspensions of the peptides described herein may be prepared as oily injection suspensions. Suitable lipophilic solvents or vehicles include fatty oils such as sesame oil, synthetic fatty acid esters such as ethyl oleate or triglycerides, or liposomes. Aqueous injection suspensions may contain substances that increase the viscosity of the suspension, such as sodium carboxymethylcellulose, sorbitol, or dextran. Suspensions may also contain suitable stabilizers or agents that increase the solubility of such peptides described herein and / or reduce aggregation of the peptides, to allow for the preparation of highly concentrated solutions. Alternatively, the peptides described herein may be lyophilized or in powder form for reconstitution with a suitable vehicle, e.g., sterile pyrogen-free water, prior to use. In some embodiments, the purified peptides are administered intravenously.

[0147] 8. Pills The peptides and compositions described herein may be formulated for oral delivery. Pharmaceutical compositions that can be used orally include tablets, push-fit capsules made of gelatin, as well as soft sealed capsules made of gelatin and plasticizers such as glycerol or sorbitol. Tablets may be made by compression or molding, optionally with one or more accessory ingredients. Compressed tablets may be prepared by compressing in a suitable machine the active ingredient in a free-flowing form such as powder or granules, optionally mixed with a binder, inert diluent, or lubricant, surface active agent, or dispersant. Molded tablets may be made by molding in a suitable machine a mixture of powdered inhibitor moistened with an inert liquid diluent. In some embodiments, the tablets are coated or scored, and the formulation for oral administration may be in a dosage suitable for such administration, formulated to provide a delayed or controlled release of the active ingredient therein. Push-fit capsules may contain the active ingredient in combination with a filler such as lactose, a binder such as starch, and / or a lubricant such as talc or magnesium stearate, and optionally a stabilizer. In soft capsules, active inhibitors can be dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin, or liquid polyethylene glycol. In some embodiments, stabilizers are added. Dragee cores are provided with suitable coatings. For this purpose, concentrated sugar solutions can be used, which can optionally contain gum arabic, talc, polyvinylpyrrolidone, carbopol gel, polyethylene glycol, and / or titanium dioxide, lacquer solutions, and suitable organic solvents or solvent mixtures. Dyes or pigments can be added to tablets or dragee coatings for identification or to characterize different combinations of active inhibitor doses.

[0148] Additional pharmaceutical approaches to targeted delivery of therapeutic agents to specific regions of the gastrointestinal tract are known. Chourasia MK,Jain SK,Pharmaceutical approaches to colon targeted drug delivery systems.,J Pharm Sci.2003 Jan-Apr; 6(1):33-66.Patel M,Shah T,Amin A.Therapeutic opportunities in colon-specific drug-delivery systems Crit Rev Ther Drug Carrier Syst.2007; 24(2):147-202.Kumar P,Mishra B.Colon targeted drug delivery systems--an overview.Curr Drug Deliv.2008 Jul; 5(3):186-98.Van den Mooter G.Colon drug delivery.Expert Opin Drug Deliv.2006 Jan; 3(1):111-25.Seth Amidon, Jack E.Brown, and Vivek S.Dave,Colon-Targeted Oral Drug Delivery Systems:Design Trends and Approaches,AAPS PharmSciTech. 2015 Aug; 16(4):731-741. Each of these references is incorporated herein by reference for such disclosure.

[0149] In addition to the ingredients specifically mentioned above, it should be understood that the inhibitors and compositions described herein may contain other agents conventional in the art that are relevant to the type of formulation in question. For example, agents suitable for oral administration may contain flavoring agents.

[0150] D. Formulation The peptides and compositions described herein may contain additional excipients to improve the therapeutic agent's performance. The compositions may contain absorption enhancers or penetration enhancers to increase the degree of absorption across epidermis or mucosal surfaces and membrane permeation. Enhancers formulated in the compositions described herein include, but are not limited to, sulfoxides such as dimethyl sulfoxide (DMSO); laurocapram (1-dodecylazacycloheptan-2-one); pyrrolidones such as n-methyl-2-pyrrolidone; terpenes and terpenoids; essential oils; oxazolidinones such as 4-decycloxazolidin-2-one; urea; cyclopentadecalactone; sodium N-[8-(2-hydroxybenzoyl)amino] These may include sodium caprylate (SNAC); 8-(N-2-hydroxy-5-chloro-benzoyl)-amino-caprylic acid (5-CNAC); medium chain fatty acids, salts, and derivatives; sodium caprate; sodium caprylate; protease inhibitors and omega-3 fatty acids; liquid mixed micellar propellants; lipid polymer micelles; alkyl glycosides; chitosan; dodecyl-2-N,N-dimethylaminopropionate (DDAIP); cell membrane lipid components; nanoparticles; liposomes, ligands; and lipophilic modifications.

[0151] The peptides and compositions described herein may be formulated as solutions, lotions, coagulants, creams, ointments, gels, foams, mucoadhesive compositions, emulsions, liposomes, coatings, cores, matrices, lyophilisates.

[0152] E. Diseases Treated The methods described herein may be used to treat or prevent tissue invasive diseases. As used herein, tissue invasive diseases include any disease, disorder, malignancy, metastasis, mutation, disease, etc., in which cells migrate, invade, metastasize, or grow in some way in a location, or grow to an extent that is pathogenic or otherwise harmful in some way to the subject in which they grow. In some embodiments, the tissue invasive disease includes endometriosis. In further embodiments, the tissue invasive disease includes cancer. The cancer may include colon cancer, squamous cell carcinoma, head and neck cancer, pancreatic cancer, breast cancer, myeloid leukemia, basal cell carcinoma, synovial sarcoma, non-small cell lung cancer, solid tumors, or prostate cancer. In further embodiments, the tissue invasive disease includes fibrosis. Fibrosis can include hepatitis A, hepatitis B, hepatitis C, adenomyosis, pulmonary fibrosis, cystic fibrosis, idiopathic pulmonary fibrosis, radiation-induced lung injury, liver cirrhosis, atrial fibrosis, endomyocardial fibrosis, old myocardial infarction, glial scar, arteriosclerosis, arthrofibrosis, Crohn's disease, Dupuytren's contracture, keloid, mediastinal fibrosis, myelofibrosis, Peyronie's disease, nephrogenic systemic fibrosis, progressive massive fibrosis, retroperitoneal fibrosis, scleroderma / systemic sclerosis, or adhesive capsulitis.

[0153] F. Coadministration The peptides and compositions described herein can be co-administered with other therapeutic agents.In the embodiments described herein, the cyclic peptide composition can be administered with drugs for treating osteoporosis, including but not limited to alendronate, ibandronate, risedronate, zoledronic acid, denosumab, calcitonin, estrogen, raloxifene, bazodoxifene, teriparatide, abaloparatide, or any combination thereof. V. Exemplary Embodiments 1.Formula X1-X2-X3-X4-X5-X6-X7-X8-X9-X 10 -X 11 -X 12 -X 13 -X 14 -X 15 -X 16X1 is M or null; X2 is S, I, G, T, A, L, or null; X3 is R, K, or null; X4 is a positively charged amino acid, citrulline, Orn, D, E, 8-aminooctanoic acid, or an amino carboxylic acid having 4 to 12 carbons; X5 is M, norleucine, Orn, D, E, K, H, R, K, 8-aminooctanoic acid, an amino carboxylic acid having 4 to 12 carbons, or null; X6 is W, Y, F, or N-methyl A; X7 is F, I, L, Chg, Cha, or Tle; X8 is L, I, or A; X9 is L, I, or A; 10 is C, S, A, Abu, C(me), or S(Bzl); 11 is F, H, A, K, E, Chg, Cng, or Orn; X 12 is W, Y, A, or F; and X 13 is G, GABA, or null. 2.Formula R-X1-X2-X3-X4-X5-X6-X7-X8-X9-X 10 -X 11 -X 12 -X 13 -X 14 -X 15 -X 16 wherein R is NH2, acetylated, stearic acid, palmitic acid, myristic acid, lauric acid, C1-C8 hydrocarbon, C1-C8 fatty acid, or null; X1 is M, G, β-alanine, norleucine, norvaline, or null; X2 is W, N-methyl-W, R, Y, F, citrulline, or K; X3 is P, W, N-methyl-W, N-ethyl-W, N-methyl-A, N-ethyl-A, L, Pip, Aib, Y, or F; X4 is E, Q, N, or D; X5 is S, α-methyl-S, K, D, Orn, T, or E; X6 is I, Chg, H, or L; X7 is L or I; X8 is D, N, E, or Q; X9 is D, E, K, Q, or Orn; 10 is H or methyl-H; X 11is V, α-methyl V, Chg, L, I, or norvaline; X 12 is Q, Aib, S, R, or N; X 13 is R, K, Citrulline, Orn, D, or E; X 14 is V, I, L, or norvaline; X 15 is W, Y, or F; and X 16 is R, G, or null. 3. A peptide comprising the amino acid sequence of any one of SEQ ID NO:1 to SEQ ID NO:500. 4. A peptide comprising an amino acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with any one of SEQ ID NO:1 to SEQ ID NO:500. 5. The peptide of embodiment 1-4, wherein the peptide binds to β-catenin. 6. The peptide of embodiment 1-4, wherein the peptide is a β-catenin inhibitor. 7. The peptide of embodiment 1-4, wherein the peptide is an inhibitor of β-catenin translocation to the nucleus. 8. The peptide of embodiment 1-4, wherein the peptide prevents β-catenin from acting as a transcription factor on oncogenes, matrix metalloproteinase 9 (MMP9), or chloride C3 channel (ClC-3). 9. The peptide of embodiment 1-4, wherein the peptide prevents EMS cell transformation, invasion, migration, fibrosis, or any combination thereof. 10. The peptide according to embodiment 1-4, wherein the peptide prevents β-catenin from binding to the estrogen receptor (ESR1). 11. The peptide according to embodiment 1-4, wherein the peptide does not affect the membrane activity of β-catenin. 12. The peptide according to embodiment 1-4, wherein the peptide does not affect β-catenin E-cadherin binding. 13. The peptide according to embodiment 1-4, wherein the peptide prevents oncogenic transcription factor activity. 14. The peptide has an EC50 of less than 50uM, less than 30uM, less than 10uM, 5uM EC50, 1uM, 500nM, 400nM, 300nM, 200nM, 100nM, 50nM, 30nM, 10nM, 5nM, 3nM, 1nM, 800pM, 600pM, 400pM, 200pM, 100pM, 50pM, 30pM, 20pM, 10pM, or 5pM, or less than about 10uM, about 5uM, about 1uM, about 500nM, about 400nM, about 300nM, about 200nM, about 100nM, about 50nM, about 30nM, about 10nM, about 5nM, about 3nM, about 1nM, about 800 EC50 of about 500 pM, about 600 pM, about 400 pM, about 200 pM, about 100 pM, about 50 pM, about 30 pM, about 20 pM, about 10 pM, about 5 pM 50and / or inhibits Wnt pathway activity with less than about 50 uM, about 30 uM, 10 uM, about 5 uM, about 1 uM, about 500 nM, about 400 nM, about 300 nM, about 200 nM, about 100 nM, about 50 nM, about 30 nM, about 10 nM, about 5 nM, about 3 nM, about 1 nM, about 800 pM, about 600 pM, about 400 pM, about 200 pM, about 100 pM, about 50 pM , about 30 pM, about 20 pM, about 10 pM, or about 5 pM, or less than 10 uM, 5 uM, 1 uM, 500 nM, 400 nM, 300 nM, 200 nM, 100 nM, 50 nM, 30 nM, 10 nM, 5 nM, 3 nM, 1 nM, 800 pM, 600 pM, 400 pM, 200 pM, 100 pM, 50 pM, 30 pM, 20 pM, 10 pM, or 5 pM 50The peptide according to embodiment 1-13, which binds to β-catenin with binding affinity. 15. The peptide according to embodiment 1-14, wherein the peptide is not naturally occurring. 16. The peptide according to embodiment 1-15, wherein the peptide is a cyclized peptide. 17. The peptide according to embodiment 1-15, wherein the peptide is a bicyclic peptide. 18. The peptide according to embodiment 16 or 17, wherein the peptide is cyclized at a Cys-Cys disulfide bond. 19. The peptide according to embodiment 16 or 17, wherein the peptide is cyclized at an amide bond. 20. The peptide according to embodiment 19, wherein the amide bond is head-to-tail between the N-terminus and the C-terminus. 21. The peptide according to embodiment 19, wherein the amide bond is head-to-side chain between the N-terminus and the internal COOH. 22. The peptide according to embodiment 19, wherein the amide bond is chain-to-tail between the internal NH2 and the C-terminus. 23. The peptide according to embodiment 19, wherein the amide bond is side chain-side chain between the internal NH2 and the internal COOH. 24. The peptide according to embodiment 16 or 17, wherein the peptide is cyclized using hydrocarbon stapling. 25. The peptide according to embodiment 16 or 17, wherein the peptide is cyclized using click chemistry.26. The peptide is at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, at least 21, at least 22, at least 23, at least 24, at least 25, at least 26, at least 27, at least 28, at least 29, at least 30, at least 31, at least 32, at least 33, at least 34, at least 35, at least 36, at least 37, at least 38, at least 39, at least 40, at least 41, at least 42, at least 43, at least 44, at least 45, at least 46, at least 47, at least 48, at least 49, at least 50, at least 51, at least 52, at least 53, at least 54, at least 55, at least 56, at least 57, at least 58, at least 59, at least 60, at least 61, at least 62, at least 63, at least 64, at least 65, at least 66, at least 67, at least 68, at least 69, at least 70, at least 71, at least 72, at least 73, at least 74, at least 75, at least 76, at least 77, at least 78, at least 79, at least 80, or at least 81 amino acid residues.27. Peptides are less than 4, less than 5, less than 6, less than 7, less than 8, less than 9, less than 10, less than 11, less than 12, less than 13, less than 14, less than 15, less than 16, less than 17, less than 18, less than 19, less than 20, less than 21, less than 22, less than 23, less than 24, less than 25, less than 26, less than 27, less than 28, less than 29, less than 30, less than 31, less than 32, less than 33, less than 34, less than 35, less than 36, less than 37, less than 38, less than 39, less than 40, less than 41, less than 42, less than 43, less than 44, less than 45, less than 46 27. The peptide of any one of embodiments 1-26, wherein the amino acid residues are less than 47, less than 48, less than 49, less than 50, less than 51, less than 52, less than 53, less than 54, less than 55, less than 56, less than 57, less than 58, less than 59, less than 60, less than 61, less than 62, less than 63, less than 64, less than 65, less than 66, less than 67, less than 68, less than 69, less than 70, less than 71, less than 72, less than 73, less than 74, less than 75, less than 76, less than 77, less than 78, less than 79, less than 80, or less than 81. 28. The peptide of any one of embodiments 1-27, wherein the peptide comprises one or more non-natural amino acids. 29. The peptide of embodiment 28, wherein the one or more non-natural amino acids are N-methyl amino acids. 30. A pharmaceutical composition comprising a β-catenin inhibitor and a pharma- ceutically acceptable carrier. 31. A pharmaceutical composition comprising a peptide that comprises binding specificity for β-catenin and a pharma- ceutically acceptable carrier. 32. A pharmaceutical composition comprising a β-catenin inhibitor comprising a peptide and a pharma- ceutical acceptable carrier. 33. A pharmaceutical composition comprising a peptide according to any one of embodiments 1-29 and a pharma- ceutical acceptable carrier. 34. A pharmaceutical composition according to any one of embodiments 30-33, wherein the peptide is a cyclized peptide. 35. A pharmaceutical composition according to embodiments 30-33, wherein said pharmaceutical composition comprises an absorption enhancer or permeation enhancer. 36. Absorption enhancers include one or more sulfoxides, such as dimethyl sulfoxide (DMSO); laurocapram (1-dodecylazacycloheptan-2-one); pyrrolidones, such as n-methyl-2-pyrrolidone; terpenes and terpenoids; essential oils; oxazolidinones, such as 4-decycloxazolidin-2-one; urea; cyclopentadecalactone; sodium N-[8-(2-hydroxybenzoyl)amino]caprylate (SNAC); 8-(N-2-hydroxy-5-chloro-benzoyl)-amino-caprylic acid (5-CNAC); medium chain fatty acids, salts, and derivatives; sodium caprate (sodium caprate). caprate); sodium caprylate; protease inhibitors and omega-3 fatty acids; liquid mixed micellar propellants; lipid polymer micelles; alkyl glycosides; chitosan; dodecyl-2-N,N-dimethylaminopropionate (DDAIP); cell membrane lipid components; nanoparticles; liposomes, ligands; and lipophilic modifications. 37. The pharmaceutical composition of embodiment 36, wherein the absorption enhancer is sodium caprate. 38. A formulation comprising a peptide according to any one of embodiments 1-29 or a pharmaceutical composition according to any one of embodiments 30-37, said formulation comprising a solution, lotion, concoct, cream, ointment, gel, foam, powder, solid, paste, tincture, microparticles, microcapsules, nanoparticles, liposomes, emulsion or lyophilisate. 39. The formulation according to embodiment 38, wherein the gel is a thermoreversible gel. 40. The formulation according to embodiment 38, wherein the formulation comprises one or more mucoadhesive polymers. 41. The formulation according to embodiment 38, wherein the formulation is for topical administration. 42. An intravaginal device comprising a peptide according to any one of embodiments 1-29 or a pharmaceutical composition according to any one of embodiments 30-37. 43. The intravaginal device of embodiment 42, wherein the intravaginal device is a suppository, a transdermal patch, a sponge, a tape, a film, a vaginal ring, a vaginal tampon, a vaginal ring, a vaginal strip, a vaginal capsule, a vaginal tablet, a vaginal pessary, a vaginal cup, a vaginal sponge, or an intrauterine device. 44. The intravaginal device of embodiment 42, wherein the peptide or the pharmaceutical composition is formulated as a formulation selected from the group consisting of a solution, a lotion, a concomitant, a cream, an ointment, a gel, a foam, a mucoadhesive composition, a coating, a core, a matrix, an emulsion, a liposome, or a lyophilisate, wherein the intravaginal device comprises the formulation. 45. The intravaginal device of embodiment 42, wherein the intravaginal device comprises from about 0.01 mg to about 5000 mg of the inhibitor. 46. ​​The intravaginal device of embodiment 42, wherein the intravaginal device comprises about 0.01 mg, about 0.05 mg, about 0.1 mg, about 0.5 mg, about 1 mg, about 5 mg, about 10 mg, about 20 mg, about 40 mg, about 60 mg, about 80 mg, about 100 mg, about 150 mg, about 200 mg, about 400 mg, about 600 mg, about 800 mg, about 1000 mg, about 1200 mg, about 1400 mg, about 1600 mg, about 1800 mg, about 2000 mg, about 2500 mg, about 3000 mg, about 3500 mg, about 4000 mg, about 4500 mg, or about 5000 mg of the inhibitor. 47. The intravaginal device of embodiment 42, wherein the intravaginal device is configured to deliver the inhibitor transmucosally.48. The above intravaginal device shall be used for the following periods: 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, 24 hours, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks 49. The intravaginal device of embodiment 42, wherein the intravaginal device is configured to deliver the inhibitor for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, 1 year, 2 years, 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, 11 years, or 12 years. 49. The intravaginal device of embodiment 42, wherein the intravaginal device is configured to deliver about 0.01 mg to about 1000 mg of the inhibitor per day. 50. The intravaginal device of embodiment 42, wherein the intravaginal device is configured to deliver about 0.01 mg, about 0.05 mg, about 0.1 mg, about 0.5 mg, about 1 mg, about 5 mg, about 10 mg, about 20 mg, about 40 mg, about 60 mg, about 80 mg, about 100 mg, about 150 mg, about 200 mg, about 400 mg, about 600 mg, about 800 mg, or about 1000 mg of the inhibitor per day. 51. The intravaginal device of embodiment 43, wherein the device is a suppository. 52. The intravaginal device of embodiment 43, wherein the device is a transdermal patch. 53. The intravaginal device of embodiment 43, wherein the device is a sponge. 54. The intravaginal device of embodiment 43, wherein the device is a tape. 55. The intravaginal device of embodiment 43, wherein the device is an intravaginal ring. 56. The intravaginal device of embodiment 55, wherein the intravaginal ring comprises a silicone insert, a compressed tablet, or a freeze-dried gel. 57. The intravaginal device of embodiment 56, wherein the peptide or the composition is incorporated throughout a silicone insert, a compressed tablet, or a freeze-dried gel. 58. The intravaginal device of embodiment 43, wherein the device is a vaginal tampon. 59. The intravaginal device of embodiment 43, wherein the device is a vaginal ring. 60. The intravaginal device of embodiment 43, wherein the device is a vaginal strip.61. The intravaginal device according to embodiment 43, wherein the device is a capsule. 62. The intravaginal device according to embodiment 43, wherein the device is a vaginal tablet. 63. The intravaginal device according to embodiment 43, wherein the device is a vaginal pessary. 64. The intravaginal device according to embodiment 43, wherein the device is a vaginal cup. 65. The intravaginal device according to embodiment 43, wherein the device is a vaginal sponge. 66. The intravaginal device according to embodiment 43, wherein the device is an intrauterine device. 67. A formulation comprising a peptide according to any one of embodiments 1-29 or a pharmaceutical composition according to any one of embodiments 30-37, wherein the formulation is for oral administration. 68. The formulation according to embodiment 67, wherein the formulation is a tablet or capsule. 69. The formulation according to embodiment 67, wherein the formulation is a liquid. 70. A method of inhibiting β-catenin in a subject, comprising inserting a device according to any one of embodiments 42-66 into the vagina of the subject. 71. A method for inhibiting β-catenin, comprising contacting a cell with a peptide according to any one of embodiments 1-29 or a pharmaceutical composition according to any one of embodiments 30-37. 72. A method for treating endometriosis, comprising administering to a subject in need thereof a therapeutically effective amount of a peptide according to any one of embodiments 1-29 or a pharmaceutical composition according to any one of embodiments 30-37. 73. A method for reducing symptoms associated with endometriosis, comprising administering to a subject in need thereof a therapeutically effective amount of a peptide according to any one of embodiments 1-29 or a pharmaceutical composition according to any one of embodiments 30-37. 74. The method of embodiment 73, wherein the symptoms include at least one selected from the group consisting of chronic pain, central sensitization, myofascial pain, adnexal mass, infertility, dysmenorrhea, genetic predisposition, non-menstrual pelvic-abdominal pain, dyspareunia, bowel symptoms (diarrhea, cramps, constipation), painful defecation (difficulty in passing stool), ovarian mass or tumor, painful bladder symptoms, and dysuria. 75. A method for treating a disease, comprising administering to a subject a therapeutically effective amount of a peptide according to any one of embodiments 1-29 or a pharmaceutical composition according to any one of embodiments 30-37. 76. The method according to embodiment 75, wherein the disease is a tissue infiltration disease, a tissue migration disease, a tissue infiltration disease, or a combination thereof. 77. The method according to embodiment 75, wherein the disease comprises cancer. 78. The method according to embodiment 77, wherein the cancer comprises colorectal cancer, squamous cell carcinoma, head and neck cancer, pancreatic cancer, breast cancer, myeloid leukemia, basal cell carcinoma, synovial sarcoma, non-small cell lung cancer, solid tumors, or prostate cancer. 79. The method according to embodiment 75, wherein the disease comprises endometriosis. 80. The method according to any one of embodiments 71-75, wherein the peptide binds to β-catenin. 81. The method according to any one of embodiments 71-80, wherein the peptide inhibits β-catenin. 82. The method according to embodiment 80, wherein the peptide binds to cytoplasmic β-catenin. 83. The method of embodiment 82, wherein the peptide binds to cytoplasmic β-catenin and inhibits translocation of β-catenin to the nucleus of the cell. 84. The method of embodiment 82, wherein the peptide binds to cytoplasmic β-catenin and reduces the amount of free nuclear β-catenin. 85. The method of embodiment 82, wherein the peptide binds to cytoplasmic β-catenin and prevents β-catenin from acting as a transcription factor on oncogenes, matrix metalloproteinase 9 (MMP9), or chloride C3 channel (ClC-3). 86. The method of embodiment 82, wherein the peptide binds to cytoplasmic β-catenin and prevents EMS cell transformation, invasion, migration, fibrosis, or any combination thereof. 87. The method of embodiment 82, wherein the peptide binds to cytoplasmic β-catenin and prevents β-catenin from binding to estrogen receptor (ESR1). 88. The method of embodiment 82, wherein the peptide binds to cytoplasmic β-catenin and does not reduce the membrane activity of β-catenin. 89. The method of embodiment 82, wherein the peptide binds to cytoplasmic β-catenin and β-catenin-E-cadherin binding is not reduced. 90. The method of embodiment 76, wherein the peptide prevents oncogenic transcription factor activity.91. The method of embodiment 84, wherein the amount of membrane β-catenin in the cells of the subject is increased by at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 100%. 92. The method of embodiment 91, wherein the increase in membrane β-catenin is relative to cells of a control subject that has not been administered a therapeutically effective amount of the pharmaceutical composition. 93. The method of embodiment 91, wherein the increase in cytoplasmic β-catenin is relative to cells of the subject obtained before the subject develops the disease. 94. The method of embodiment 91, wherein the increase in membrane β-catenin is relative to cells of the subject obtained at a different time point. 95. The method of any one of embodiments 72-94, wherein the therapeutically effective amount is from about 0.01 mg to about 1000 mg. 96. A method for preventing a disease, comprising administering to a subject a therapeutically effective amount of a peptide according to any one of embodiments 1-29 or a pharmaceutical composition according to any one of embodiments 30-37. 97. The method according to embodiment 96, wherein the disease is a tissue infiltration disease, a tissue migration disease, a tissue infiltration disease, or a combination thereof. 98. The method according to embodiment 96, wherein the disease comprises cancer. 99. The method according to embodiment 98, wherein the cancer comprises colorectal cancer, squamous cell carcinoma, head and neck cancer, pancreatic cancer, breast cancer, myeloid leukemia, basal cell carcinoma, synovial sarcoma, non-small cell lung cancer, solid tumors, or prostate cancer. 100. The method according to embodiment 98, wherein the disease comprises endometriosis. 101. The method according to any one of embodiments 96-100, wherein the peptide binds to β-catenin. 102. The method according to any one of embodiments 96-101, wherein the peptide inhibits β-catenin. 103. The method according to embodiment 101, wherein the peptide binds to cytoplasmic β-catenin. 104. The method of embodiment 103, wherein the peptide binds to cytoplasmic β-catenin and inhibits translocation of β-catenin to the nucleus of the cell. 105. The method of embodiment 103, wherein the peptide binds to cytoplasmic β-catenin and reduces the amount of free nuclear β-catenin. 106. The method of embodiment 103, wherein the peptide binds to cytoplasmic β-catenin and prevents β-catenin from acting as a transcription factor on oncogenes, matrix metalloproteinase 9 (MMP9), or chloride C3 channel (ClC-3). 107. The method of embodiment 103, wherein the peptide prevents EMS cell transformation, invasion, migration, fibrosis, or any combination thereof. 108. The method of embodiment 103, wherein the peptide prevents β-catenin from binding to estrogen receptor (ESR1). 109. The method of embodiment 103, wherein the peptide binds to cytoplasmic β-catenin and does not reduce membrane activity of β-catenin. 110. The method of embodiment 103, wherein the peptide binds to cytoplasmic β-catenin and β-catenin-E-cadherin binding is not affected. 111. The method of embodiment 96, wherein the peptide prevents oncogenic transcription factor activity.112. The method of embodiment 105, wherein the amount of free nuclear β-catenin in the cells of the subject is reduced by at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 100%. 113. The method of embodiment 112, wherein the reduction in free cytoplasmic β-catenin is relative to cells of a control subject that has not been administered a therapeutically effective amount of the pharmaceutical composition. 114. The method of embodiment 112, wherein the reduction in free nuclear β-catenin is relative to cells of the subject obtained before the subject develops endometriosis or tissue invasive disease. 115. The method of embodiment 112, wherein the reduction in free cytoplasmic β-catenin is for cells of the subject obtained at different time points. 116. The method of any one of embodiments 96-115, wherein the therapeutically effective amount is from about 0.01 mg to about 1000 mg. 117. The method of any one of embodiments 72-116, wherein the peptide or pharmaceutical composition is administered intravenously. 118. The method of any one of embodiments 72-116, wherein the peptide or pharmaceutical composition is administered intramuscularly. 119. The method of any one of embodiments 72-116, wherein the peptide or pharmaceutical composition is administered simultaneously with the administration of a drug for treating osteoporosis. 120. The method of embodiment 119, wherein the drug for treating osteoporosis comprises alendronate, ibandronate, risedronate, zoledronic acid, denosumab, calcitonin, estrogen, raloxifene, bazodoxifene, teriparatide, abaloparatide, or any combination thereof. 121. The method of embodiment 119, wherein the drug for treating osteoporosis is zoledronic acid.

[0154] VI. Working Examples The following examples are included for illustrative purposes only and are not intended to limit the scope of the disclosure.

[0155] Example 1 Peptide synthesis Antisense Met-X1-X2-X3-X4-X5-X6-X7-X8-X9-X 10 The -Lys single stranded DNA template was synthesized at the Keck Oligonucleotide Synthesis Facility (Yale) with the sequence 5'...GCCAGACCCCGATTTSNNSNNSNNSNNSNNSNNSNNSNNSNNSNNCATTGTAATTGTAAATA-GTAATTG...3', where N=A,T,C,G and S=G,C. The reagent bottle used for the "N" position was made by mixing A:C:G:T in a 3:3:2:2 ratio. The reagent bottle for the "S" position was made by mixing C:G in a 3:2 ratio. The forward primer Gen-FP (5'-TAATACGACTCACTATAGGGACAATTACTATTTACAATTA CA-3') and reverse primer MK 10 MX10K library double-stranded DNA was amplified by five cycles of polymerase chain reaction (PCR) using K-RP (5'...ACCGCTGCCAGACCCCGATT T...3'). Round 0 mRNA pools were generated by T7 runoff transcription and purified by urea-PAGE. Purified mRNA was ligated with oligonucleotide splints (MX 10 K-splint: 5'...TTTTTTTTTTTTTTACCGCTGCCAGAC...3') was ligated to F30P ((5'-dA21[C9]3dAdCdC-P; C9 = tri-(ethylene glycol) phosphate (Glen Research), P = puromycin (Glen Research). After PAGE purification of the ligation reaction, the template was dissolved in water and quantified by absorbance at 260 nm.

[0156] Example 2 Peptide synthesis and cyclization Increased protease resistance has been shown in peptides containing N-methyl amino acids. Furthermore, cyclization of peptides has been shown to increase the binding affinity of ligand peptides.

[0157] All solvents were purchased from Sigma. All peptides except N-Me-L14 were synthesized by manual solid-phase peptide synthesis using preloaded leu-2-chlorotrityl resin (250 mg, 0.15 mmol; Anaspec). N-Me-L14 was synthesized after loading Fmoc-N-methylleucine (Fmoc = 9-fluorenylmethoxycarbonyl; Anaspec) onto 2-chlorotrityl resin (Anaspec). Five N-terminal Asn residues were added to enhance peptide solubility. Loading was achieved by adding FmocN-Me-L14 in DMF with N,N-diisopropylethylamine (DIEA; 5 equiv.) to 2-chlorotrityl resin (5 equiv.; 250 mg, 0.35 mmol) for 2 h. Standard couplings were performed with monomer (5 equiv; Novabiochem) in HATU (2 mL, 0.6 mmol; Novabiochem), HOAt (1.2 mmol; Genescript) in DMF with DIEA (1.8 mmol) for 15 min at room temperature. Coupling to N-methyl amino acids followed the same procedure with a coupling time of 30 min. Fmoc deprotection was performed with 20% piperidine (Anaspec) for 15 min at room temperature. Following deprotection, cleavage with 95% TFA, filtration, and ether extraction, the crude product was purified on a Vydac C-18 reversed-phase column using gradient elution (0% B in 5 min, 10–50% B in 40 min. Solvent A: HO with 0.1% TFA, Solvent B: CHCN with 0.035% TFA). The lyophilized solid was reconstituted in DMSO and quantified by absorbance at 280 nm (ε280=12490 L mol-1 cm-1). Yield=20-25%.

[0158] The peptide was cyclized by adding 50 μL of DSG (1 mg / mL in DMF) to 190 μL of dT-purified fusion in 50 mM phosphate buffer (pH=8). The reaction was allowed to proceed for 1 h, and the fusion was repurified by dT-cellulose and ethanol precipitation. This process yielded the cyclic peptide.

[0159] Example 3 Nuclear β-catenin inhibition and Wnt signaling Peptides were evaluated for their ability to inhibit Wnt signaling. Mouse 3T3 fibroblasts expressing luciferase under a Wnt-responsive promoter (TCF / LEF) (Enzo Life Sciences, Farmingdale, NY) were stimulated with Wnt3a in the presence of increasing concentrations of peptide inhibitors (Hit1: SEQ ID NO:217, Hit2: SEQ ID NO:224, Hit3: SEQ ID NO:228, Hit4: SEQ ID NO:227, Hit5: SEQ ID NO:220, Hit6: SEQ ID NO:168. Hit7: SEQ ID NO:226) or vehicle control (Figure 2A). Optimized compounds had ECs ranging from 500 to 999 nM. 50 Values ​​are shown and luciferase levels were reduced to that of the unstimulated (-Wnt3a) control. Luciferase activity was not affected by the addition of a scrambled peptide (containing the same amino acids in a scrambled manner), a non-functional peptide (peptide does not target β-catenin), or a non-permeabilizing peptide (peptide without permeabilizing enhancing features). Results show concentration-dependent repression of Wnt-responsive promoters with the peptides described herein. conc dep supp.

[0160] Example 4 The peptides do not inhibit membrane-bound β-catenin or block access to its binding site on E-cadherin. A competitive binding assay was performed (Figure 3), in which biotinylated β-catenin was immobilized on a streptavidin plate and 50 nM of E-cadherin FC fusion was added in the presence of various amounts of peptide inhibitor (SEQ ID NO: 42). None of the doses tested inhibited the interaction of β-catenin with E-cadherin. The results show that the peptides do not inhibit membrane-bound β-catenin or block access to the binding site on E-cadherin.

[0161] Example 5 β-catenin inhibitors induce lesion regression Studies were performed to evaluate the impact of β-catenin on a mouse model of endometriosis. Lesions were established in the EMS mouse model of endometriosis by intraperitoneal injection of minced GFP+ uterine tissue from mice into wild-type c57 / bl6 mice. Lesions in this mouse model mimic the attachment sites seen in humans. The lesions are organized with defined epithelial glandular structures, organized stroma, and hemosiderin-laden macrophages. The lesions are hormonally responsive and express β-catenin, MMPs, Wnt4, PGR, and ESR. Endometriotic disease was established in mice and allowed to progress for 3 weeks before initiation of treatment.

[0162] After this establishment phase, mice were treated with either vehicle (PBS + 0.5% DMSO negative control) or the macrocyclic peptide cyclo-acetyl-K * Nle-W-3-Cyclohexyl-L-alanine-Li-(aminobutyric acid)-AWD *Mice were treated IP daily for 3 weeks with either "Peptide 2" (PBS + 5 mg / kg Peptide 2 in 0.5% DMSO) or "Peptide 2" (PBS + 5 mg / kg Peptide 2 in 0.5% DMSO). Vaginal smears were taken daily, which showed that mice treated with either vehicle or Peptide 2 progressed through the four stages of the estrous cycle. Figure 4A shows that mice cycle through proestrus (P), estrus (E), diestrus (D), and metestrus (M) stages every 4-5 days. The estrus pattern for vehicle and Peptide 2 is normal. No adverse effects on the uterus of the hypothalamic gonadal axis were noted.

[0163] Mice were then euthanized during estrus; mouse weights, ovarian weights, and uterine weights were measured and determined to be consistent across treatment regimens (Figure 4B). Compared to vehicle alone, EMS lesions were reduced in size in mice treated with peptide 2 (Figure 4B and Figure 4C). An associated increase in activated macrophages was also observed in mice treated with peptide 2 compared to vehicle alone. Figure 5 shows an increase in the relative numbers of activated macrophages, foam cells, polymorphonuclear neutrophils, B cells, and T cells at the site of lesions treated with peptide 2 (SEQ ID NO: 215) compared to vehicle alone, as assessed by histology. Without wishing to be limited by theory, these results may suggest that immune cells are phagocytosing uterine tissue.

[0164] From this experiment it can be concluded that peptide 2, an inhibitor of β-catenin, causes regression of lesions without altering estrous cycle, mouse weight, ovary weight, or uterine weight. Together, these data demonstrate that peptide 2 (1) is membrane permeable and interferes with the nuclear transcription function of β-catenin in a dose-dependent manner, (2) exhibits protease stability and a sustained half-life, (3) does not affect β-catenin membrane binding to E-cadherin, (4) inhibits cell proliferation in cells with pathological and abnormal Wnt-dependent proliferation, (5) does not affect proliferation or interfere with membrane-bound β-catenin activity in normal cells, and (6) causes regression of EMS lesions without altering uterine cycle or uterine weight.

[0165] Example 6 Therapeutic Efficacy of Peptides Peptide inhibition is tested for its ability to inhibit cell proliferation across a panel of cell types, including human endometrial stromal cells (HESC) and / or primary EMS cells from various lesion subtypes (based on location as well as severity). Mouse uterine cells derived from mouse uterus are used as a control cell line to demonstrate that the peptides do not inhibit normal cell behavior. Primary EMS cells were excised from various patients and harvested according to the World Endometriosis Research Foundation Endometriosis Phenome and Biobanking Harmonization Project (EPHect) standard protocols. All cell lines were plated (5-10x10 in quadruplicates). 4 Cells / 24-well plate), each peptide is administered daily for 5 days at 10 different concentrations (10-50,000 nM) along with a series of controls (see Table 2). After the 5th day, viable cells are detected using the CellTiter-Glo® Luminescent Cell Viability Assay (Promega). IC50 values ​​are graphed and calculated using Prism software (GraphPad) based on the cell viability readings.

[0166] [Table 2]

[0167] Example 7 Therapeutic inhibition of cell migration and invasion in vitro Cell migration and invasion in vitro of HESC, mouse EMS lesion cells, and human EMS lesion cells are analyzed using uncoated or matrigel-coated 24-well chambers or microfilters (Becton Dickinson), respectively, as reviewed by Matsuzaki et al., C. PLoS One 8, e61690. Upon stimulation and subsequent administration of the compounds determined to be optimal in Examples 3 and 6 using the same controls, cell motility and migration are calculated by the number of cells migrating into the unpopulated area of ​​the chamber in 24 hours, and cell invasion is calculated by the number of cells invading the micropore filter in 24 hours. These experiments are performed in quadruplicate and quantitatively analyzed using computerized light microscopy. The results are used to evaluate the ability of peptides to inhibit cell migration and invasion.

[0168] Example 8 Confirmation of on-target activity of therapeutic molecules in cell lines The on-target activity of each peptide was assessed using human WNT signaling pathway RT 2Transcriptional activity of >80 WNT pathway genes is assessed using Profiler PCR arrays (Qiagen, Venlo, Netherlands). Primary cells are treated with 500 nM of selected peptides for 48 h. RNA is extracted from harvested cells and profiled on the array using rtPCR. The array contains 84 genes related to Wnt-mediated signaling, including ligands, receptors, downstream signaling molecules, and regulators of the Wnt canonical pathway, two non-canonical pathways, planar polarity, and calcium ion-dependent pathways. Western blot analysis of ClC-3 and zymography of MMP-9 activity are used to confirm that selected proteins are reduced by ≥50% of initial levels. Cells are stained for β-catenin localization by immunofluorescence. Controls are as listed in Table 2.

[0169] Example 9 Therapeutic regression of lesions in vivo A mouse model of EMS is used to select compounds for further testing of therapeutic potential. EMS is induced in mice according to Burns,et al.,Endocrinology 153,3960-3971. Hormonally intact syngeneic mice are synchronized with pregnant mare serum gonadotropin (PMSG-5 IU). The uterus is removed 41 hours later, at which time hormone levels and vaginal histology mimic metestrus / diestrus (data not shown). The outer myometrium and attached blood supply are stripped and the uterus is cut longitudinally into strips, chopped into 1-2 mm pieces and placed in sterile saline. The chopped uterine tissue is injected into the peritoneal cavity through a 5 mm dorsal hole (day 0). The chopped uterine tissue is allowed to attach and grow for 3 weeks before introducing peptides into the peritoneal cavity. Mice are treated daily for 6 weeks with 5 mg / kg, 10 mg / kg, and 20 mg / kg of peptides. Mice are euthanized at estrus 9 weeks after the initiation of EMS, with previous data used as preliminary estimates of mean and standard deviation to determine numbers.

[0170] At necropsy, peritoneal cells and fluids are harvested by peritoneal lavage. Lesions are removed with surrounding tissue for histology / immunohistochemistry (IHC) or without surrounding tissue for gene expression analysis by real-time PCR (weight, volume, and location are recorded). To visualize blood vessels histologically, mice are injected with lectin (100 μg) and heparin sulfate (100 U) 20 min prior to necropsy. Lesions are analyzed histologically for vascular density, organization / collagen (Masson's Trichrome), leukocyte infiltration (i.e., neutrophils by Ly6G, macrophages by F4 / 80), proliferation (Ki67), β-catenin localization, and Wnt signaling molecules. Lesions are further used to examine Wnt / β-catenin levels, and matrix metalloproteinase (MMP) activity using MMP zymography. Ascites cells are analyzed for macrophages and neutrophils. Lesions are analyzed using real-time PCR targeting the most affected genes in the Wnt signaling PCR array in Example 4. Control-treated lesions serve as baseline.

[0171] Compounds that reduce Wnt signaling, reduce lesion weight or volume, reduce MMP activity levels, reduce fibrosis, increase leukocyte infiltration in lesions, reduce the amount of nuclear β-catenin, or any combination thereof, compared to controls, were selected for further testing.

[0172] [Table 3]

[0173] Example 10 Immunohistochemistry of localization Endometriosis was established in a mouse model of disease. Mice were allowed to establish endometriosis for 3 weeks. After the first 3 weeks, mice were intraperitoneally treated daily for 3 weeks with therapeutic peptides corresponding to SEQ ID NO: 215 or 393, or with an untreated negative control. To evaluate peptide and β-catenin localization, the lesions of the mouse model of endometriosis were evaluated. On the last day of treatment, mice were dosed with biotinylated tagged therapeutic peptides or controls to visualize peptide localization relative to the lesions. Mice were euthanized, and lesions were removed and fixed in formalin for histological and immunohistochemical examination. Fixed and sectioned lesions were incubated with specific antibodies specific for β-catenin (Cell Signaling, 1:100, secondary: Fisher Scientific, goat-anti-rabbit-647 1:200), streptavidin for biotinylated therapeutics (Fisher Scientific, SA-405 1:100), and Sytox green for nuclei (Fisher Scientific, 1:300). Immunohistochemical negative controls were performed on lesion sections from all treatment groups using an IgG non-specific antibody (rabbit-IgG, AbCam 1:100).

[0174] Qualitatively, as shown in Figure 6A, it is clear that the targeted β-catenin is not localized to the nucleus when treated with the therapeutic peptide. In addition, the therapeutic peptide further follows the localization pattern of β-catenin, which is decreased in the nucleus and maintained / increased in the membrane. This suggests that the peptide is membrane soluble and binds to cytoplasmic β-catenin and does not interfere with the binding between β-catenin in the membrane. Therapeutic peptide binding may increase the binding affinity of β-catenin to E-cadherin. Conversely, control-treated lesions had more diffuse staining across the membrane, cytosol, and nucleus. Furthermore, as shown in Figure 6B, orthogonal slices show that β-catenin in the peptide-treated lesions is more membranous compared to the control-treated lesions. As expected, the localization of the therapeutic peptide correlates with β-catenin staining. In peptide-treated lesions, peptide localization was visualized predominantly throughout the membrane.

[0175] Quantification of β-catenin localization was performed using Zeiss Blue software via thresholding, which allows the software to calculate the stained area using pixel intensity; thus, quantitative values ​​for staining can be estimated for various cellular compartments (see Table 6).

[0176] [Table 4]

[0177] Compiled area thresholding data indicates that β-catenin from lesions undergoing therapeutic treatment is more membrane-associated when compared to control-treated lesions (see FIG. 7A). Furthermore, nuclear β-catenin levels are decreased in lesions treated with therapeutic agents compared to control-treated lesions (see FIG. 7B).

[0178] Together, these data indicate that the therapeutic peptide maintains or increases membrane binding of β-catenin and inhibits nuclear β-catenin localization.

[0179] Example 11 KD50 measurement using surface plasmon resonance (SPR) Biotinylated protein (β-catenin) was immobilized on a high affinity streptavidin (SA) sensor chip (GE). Increasing concentrations of peptide were run in HBS EP+ buffer at room temperature according to the manufacturer's instructions. The maximum binding capacity of the surface (response at saturation) was calculated using the Biacore® system (GE) instructions, and binding was measured at each concentration as a data point. Binding levels were determined over a range of analyte concentrations from at least 20% to 80% saturation of the surface. On-rates, off-rates, and KD were calculated using Biacore® Insight Evaluation Software software (GE). The concentration at 50% saturation was calculated from these dilution series and is expressed as KD50 in Table 5. On-target activity of the therapeutic molecules in the cell lines was confirmed by qPCR.

[0180] The on-target activity of each peptide was assessed using Qiagen's human WNT pathway RT 2 The transcriptional activity of WNT pathway genes was assessed using the Profiler PCR array. Primary cells were treated with a therapeutic peptide corresponding to SEQ ID NO:42 for 48 hours. RNA was extracted from harvested cells and profiled on the array using rtPCR. The array contains 84 genes related to Wnt-mediated signaling, including ligands, receptors, downstream signaling molecules, and regulators of the Wnt canonical pathway, two non-canonical pathways, planar polarity, and calcium ion-dependent pathways, of which 38 were evaluated here. A positive control of ICG001 (a known Wnt inhibitor) was also run on the RNA array and used as a comparison.

[0181] The results shown in Figures 8A and 8B indicate that the therapeutic peptide corresponding to SEQ ID NO:42 outperformed known positive controls by inhibiting several key gene transcripts in the Wnt pathway. Specifically, when the therapeutic peptide was administered, downstream Wnt markers and target genes were downregulated, including AXIN1, AXIN2, CCND1, CTNNB1, CXXC4, DKK1, DKK3, DVL1, DVL2, FRAT1, LEF1, MMP7, MYC, NFATC1, RHOA, RHOU, TCF7, TCF7L1, WNT9A, and Wnt receptors and ligands, including FZD3, FZD4, FZD6, FZD8, WNT10A, WANT11, WANT2B, WNT4, WNT5A, WNT5B, WNT6, WNT7A (see Figures 8A and 8B). Downregulation of important transcripts may contribute to the anti-proliferative effect of therapeutic agents observed in endometriosis cells.In addition, some Wnt receptors and ligands are upregulated, which may be the result of apoptosis in these treated cells.Upregulated Wnt receptors and ligands include FZD5, FZD9, WNT3.Together, these results indicate that therapeutic peptide corresponding to SEQ ID NO:42 is a potent and specific Wnt inhibitor.

[0182] Example 12 Measurement of KD50 using Fluorescence Polarization (FP) 5 nM of labeled TCF4 protein was added to a solution of 100 nM β-catenin and incubated at room temperature in a 96-well plate (Greiner96 flat clear plate, Millipore Sigma (Carlsbad CA)). Various concentrations of a β-catenin inhibitor candidate (SEQ ID NO:404) were then added to a series of serial dilutions of the inhibitor from 10 nM to 30 uM in different wells of the plate. Fluorescence polarization was read by an Infinite M1000 (Tecan, Baldwin Park CA) at an excitation wavelength of 470 nm and measured at an emission wavelength of 520 nm. Polarization measurements corrected for background noise from the blank resulted in a reading of the polarization intensity for each data point. Experiments were performed in triplicate and data were analyzed and plotted in Microsoft Excel (Microsoft, Redmond, WA). An example showing the results of SEQ ID NO:404 is shown in Figure 9. The above results indicate that the inhibitor binds to β-catenin in a concentration-dependent manner.

[0183] Example 13 Effect of peptides on cell proliferation Peptide inhibitors were tested for their ability to inhibit cell proliferation across two different β-catenin expressing pathogenic cell lines: a colon cancer cell line (SW480) derived from a mouse model of the disease and / or mouse EMS lesion cells. Both cell lines are known to express high levels of β-catenin. Cell lines were incubated with various peptide concentrations and assessed for viability after 5 and 7 days. Viable cells were detected using the CellTiter-Glo® Luminescent Cell Viability Assay (Promega). IC50 values ​​are graphed and calculated using Prism software (GraphPad) based on cell viability readings. IC50 is used to assess inhibition of cell proliferation by peptide concentration and calculate the inhibitory effect of each peptide across each line. The results are used to evaluate the therapeutic efficacy of the peptides.

[0184] In SW480 cell line, peptide inhibitors from family 1 (SEQ ID NO:264) and family 2 (SEQ ID NO:396) were added daily to cell growth medium. After 7 days, cell viability was assessed using Cell-Titer Glo kit (Promega, Madison, WI). Both peptides inhibited cell growth after 7 days in a concentration-dependent manner (Figure 2B). In mouse EMS lesion cells derived from disease mouse model, peptide inhibitor (SEQ ID NO:465) was added daily to cell growth medium at concentrations of 25uM, 12.5uM, 6.25uM, 3.125uM, 1.56uM, 0.78uM, 0.39uM, 0.195uM, 0.098uM, 0.049uM, 0.024uM, and 0uM (negative control). Cell viability was assessed after 5 days, and peptides inhibited cell growth after 5 days in a dose-dependent manner. IC 50 was calculated for peptides based on the concentration at which 50% viability was observed (Table 5).

[0185] Example 14 Exemplary Compounds Exemplary peptides are shown below: Peptides were synthesized as described in Examples 1 and 2.

[0186] [ka]

[0187] [ka]

[0188] [ka]

[0189] [ka]

[0190] [ka]

[0191] While preferred embodiments of the present disclosure have been shown and described herein, it will be apparent to those skilled in the art that such embodiments are provided by way of example only. Numerous variations, changes, and substitutions will occur to those skilled in the art without departing from the present disclosure. It is understood that various alternatives to the embodiments of the disclosure described herein may be utilized in the practice of the present disclosure. The following claims define the scope of the disclosure, and it is intended that methods and structures within the scope of the claims and their equivalents be covered thereby.

[0192] [Table 5-1]

[0193] [Table 5-2]

[0194] [Table 5-3]

[0195] [Table 5-4]

[0196] [Table 5-5]

[0197] [Table 5-6]

[0198] [Table 5-7]

[0199]

Table 5-8

[0200]

Table 5-9

[0201]

Table 5-10

[0202]

Table 5-11

[0203]

Table 5-12

[0204]

Table 5-13

[0205]

Table 6-1

[0206]

Table 6-2

[0207]

Table 6-3

[0208]

Table 6-4

[0209]

Table 6-5

[0210]

Table 6-6

[0211]

Table 6-7

[0212]

Table 6-8

[0213]

Table 6-9

[0214]

Table 6-10

Claims

1. Formula X １ -X ２ -X ３ -X ４ -X ５ -X ６ -X ７ -X ８ -X ９ -X １０ -X １１ -X １２ -X １３ -X １４ -X １５ -X １６ A peptide comprising an amino acid sequence having the formula: X １ is M or null, X ２ is S, I, G, T, A, L, or null; X ３ is R, K, or null; X ４ is a positively charged amino acid, citrulline, Orn, D, E, 8-aminooctanoic acid, or an aminocarboxylic acid having 4-12 carbons; X ５ is M, norleucine, Orn, D, E, K, H, R, K, 8-aminooctanoic acid, an aminocarboxylic acid having 4 to 12 carbons, or null; X ６ is W, Y, F, or N-methyl A; X ７ is F, I, L, Chg, Cha, or Tle; X ８ is L, I, or A; X ９ is L, I, or A; X １０ is C, S, A, Abu, C(me), or S(Bzl); X １１ is F, H, A, K, E, Chg, Cng, or Orn; X １２ is W, Y, A, or F, and X １３ is G, GABA, or null; peptide.

2. Formula RX １ -X ２ -X ３ -X ４ -X ５ -X ６ -X ７ -X ８ -X ９ -X １０ -X １１ -X １２ -X １３ -X １４ -X １５ -X １６ A peptide comprising an amino acid sequence having the formula: R is NH ２ , acetylated, stearic acid, palmitic acid, myristic acid, lauric acid, C １ -C ８ Hydrocarbons, C １ -C ８ fatty acid, or null, X １ is M, G, β-alanine, norleucine, norvaline, or null; X ２ is W, N-methyl W, R, Y, F, citrulline, or K; X ３ is P, W, N-methyl-W, N-ethyl-W, N-methyl A, N-ethyl A, L, Pip, Aib, Y, or F; X ４ is E, Q, N, or D; X ５ is S, alpha methyl S, K, D, Orn, T, or E; X ６ is I, Chg, H, or L; X ７ is L or I, X ８ is D, N, E, or Q; X ９ is D, E, K, Q, or Orn; X １０ is H or methyl-H, X １１ is V, alpha methyl V, Chg, L, I, or norvaline; X １２ is Q, A, S, R, or N; X １３ is R, K, Citrulline, Orn, D, or E; X １４ is V, I, L, or norvaline; X １５ is W, Y, or F, and X １６ is R, G, or null; peptide.

3. The peptide according to claim 1 or 2, comprising any one of the amino acid sequences of SEQ ID NO: 1 to SEQ ID NO:

500.

4. 3. The peptide of claim 1 or 2, comprising an amino acid sequence having at least 80% sequence identity with any one of SEQ ID NO: 1 to SEQ ID NO:

500.

5. The peptide of claim 1 or 2, wherein the peptide binds to β-catenin.

6. The peptide of claim 1 or 2, wherein the peptide is a β-catenin inhibitor.

7. The peptide according to claim 1 or 2, which is an inhibitor of β-catenin translocation to the nucleus.

8. 3. The peptide of claim 1 or 2, wherein the peptide prevents β-catenin from acting as a transcription factor on oncogenes, matrix metalloproteinase 9 (MMP9), or chloride C3 channel (ClC-3).

9. 3. The peptide of claim 1 or 2, wherein the peptide prevents endometriotic (EMS) cell transformation, invasion, migration, fibrosis, or any combination thereof.

10. The peptide of claim 1 or 2, wherein the peptide prevents β-catenin from binding to the estrogen receptor (ESR1).

11. The peptide of claim 1 or 2, wherein the peptide does not reduce the membrane activity of β-catenin.

12. The peptide of claim 1 or 2, wherein the peptide does not reduce β-catenin E-cadherin binding.

13. The peptide of claim 1 or 2, wherein the peptide prevents oncogenic transcription factor activity.

14. The peptide has an EC ５０ The peptide of claim 5, which inhibits Wnt pathway activity having the formula:

15. The peptide of claim 1 or 2, wherein the peptide is not naturally occurring.

16. 3. The peptide of claim 1 or 2, wherein the peptide is a cyclized peptide or a bicyclic peptide.

17. The peptide of claim 16, wherein the peptide is cyclized at a Cys-Cys disulfide bond.

18. 17. The peptide of claim 16, wherein the peptide is cyclized at an amide bond.

19. 19. The peptide of claim 18, wherein the amide bond is head-to-tail between the N-terminus and C-terminus.

20. 19. The peptide of claim 18, wherein the amide bond is head-to-side chain between the N-terminus and an internal COOH.

21. The amide bond is an internal NH ２ and the C-terminus-tail.

22. The amide bond is an internal NH ２ and an internal COOH, side chain-side chain.

23. 17. The peptide of claim 16, wherein the peptide is cyclized using hydrocarbon stapling.

24. The peptide of claim 16, wherein the peptide is cyclized using click chemistry.

25. 3. The peptide of claim 1 or 2, wherein the peptide is at least three amino acid residues.

26. 3. The peptide of claim 1 or 2, wherein the peptide is less than 81 amino acid residues.

27. The peptide of claim 1 or 2, wherein the peptide comprises one or more unnatural amino acids.

28. 28. The peptide of claim 27, wherein the one or more unnatural amino acids are N-methyl amino acids.

29. A pharmaceutical composition, comprising: β-catenin inhibitors; A peptide that contains binding specificity for β-catenin; a peptide-containing β-catenin inhibitor; or A peptide comprising any one of the amino acid sequences of SEQ ID NOs: 1-490; and including a pharma- ceutically acceptable carrier, Pharmaceutical compositions.

30. A method for treating endometriosis, comprising administering to a subject in need thereof a therapeutically effective amount of a peptide comprising an amino acid sequence set forth in any one of SEQ ID NOs: 1-490 or the pharmaceutical composition of claim 29.

31. 31. The method of claim 30, wherein treating endometriosis reduces symptoms associated with endometriosis, said symptoms including at least one selected from the group consisting of chronic pain, central sensitization, myofascial pain, adnexal mass, infertility, dysmenorrhea, genetic predisposition, non-menstrual pelvic-abdominal pain, dyspareunia, bowel symptoms (diarrhea, cramps, constipation), painful defecation (difficulty in passing stool), ovarian mass or tumor, painful bladder symptoms, and dysuria.

32. The method of claim 30, wherein the peptide binds to cytoplasmic β-catenin.

33. The method of claim 30, wherein the peptide inhibits β-catenin.

34. The method of claim 32, wherein the peptide binds to cytoplasmic β-catenin and inhibits translocation of β-catenin to the nucleus of the cell.

35. The method of claim 32, wherein the peptide binds to cytoplasmic β-catenin and maintains or increases membrane-bound β-catenin.

36. 33. The method of claim 32, wherein the peptide binds to cytoplasmic β-catenin and prevents β-catenin from acting as a transcription factor on oncogenes, matrix metalloproteinase 9 (MMP9), or chloride C3 channel (ClC-3).

37. 33. The method of claim 32, wherein the peptide binds to cytoplasmic β-catenin and prevents EMS cell transformation, invasion, migration, fibrosis, or any combination thereof.

38. The method of claim 32, wherein the peptide binds to cytoplasmic β-catenin and prevents β-catenin from binding to the estrogen receptor (ESR1).

39. The method of claim 32, wherein the peptide binds to cytoplasmic β-catenin and does not reduce the membrane activity of β-catenin.

40. The method of claim 32, wherein the peptide binds to cytoplasmic β-catenin and does not reduce β-catenin-E-cadherin binding.

41. 31. The method of claim 30, wherein the peptide prevents oncogenic transcription factor activity.

42. The method of claim 32, wherein the amount of nuclear β-catenin in the subject's cells is reduced by at least 5%.

43. The method of claim 42, wherein the reduction in nuclear β-catenin is relative to cells of a control subject that has not been administered a therapeutically effective amount of the pharmaceutical composition.

44. The method of claim 42, wherein the reduction in nuclear β-catenin is for cells of the subject obtained before the subject develops the disease.

45. The method of claim 42, wherein the reduction in nuclear β-catenin is for cells of the subject obtained at different time points.

46. 31. The method of claim 30, wherein the therapeutically effective amount is from about 0.01 mg to about 1000 mg.

47. The method of claim 30, wherein the peptide or pharmaceutical composition is administered intravenously.

48. The method of claim 30, wherein the peptide or pharmaceutical composition is administered intramuscularly.

49. The method of claim 30, wherein the peptide or the pharmaceutical composition is administered simultaneously with the administration of a drug for treating osteoporosis.

50. 50. The method of claim 49, wherein the drug for treating osteoporosis comprises alendronate, ibandronate, risedronate, zoledronic acid, denosumab, calcitonin, estrogen, raloxifene, bazodoxifene, teriparatide, abaloparatide, or any combination thereof.

51. 50. The method of claim 49, wherein the drug for treating osteoporosis is zoledronic acid.

52. An intravaginal device comprising a peptide comprising the amino acid sequence of SEQ ID NO: 215 or 393 formulated to release the peptide in a time-controlled manner, wherein the peptide binds to β-catenin; wherein the peptide inhibits translocation of β-catenin to the nucleus of a cell; wherein the peptide does not reduce β-catenin binding to E-cadherin in the membrane; and wherein the peptide reduces the size or severity of endometriotic lesions or other symptoms associated with endometriosis in a subject in need thereof. Intravaginal device.

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