Compositions and methods for imaging and treating endometriosis

Abstract

Provided herein are peptides that bind β-catenin, and compositions comprising said peptides. Agents utilizing peptides binding directly to β-catenin can be used for imaging, diagnostic, and therapeutic purposes of diseases that are driven by β-catenin dysfunction. Also provided herein are methods of using said peptides and compositions in the imaging, diagnosis, and treatment of tissue-infiltrating conditions including endometriosis.

Description

COMPOSITIONS AND METHODS FOR IMAGING AND TREATING ENDOMETRIOSISCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims benefit of U.S. Provisional Application No. 63 / 504,185, filed May 24, 2023, which is hereby incorporated herein by reference in its entirety. STATEMENT OF GOVERNMENT INTEREST

[0002] This invention was made with Government Support under Grant No. HD100289 awarded by the National Institutes of Health. The Government has certain rights in the invention.SEQUENCE LISTING

[0003] This application contains a sequence listing filed in ST.26 format entitled “170502-2010 Sequence Listing” created on May 24, 2024, having 566,033 bytes. The content of the sequence listing is incorporated herein in its entirety.BACKGROUND

[0004] Endometriosis (EMS) affects approximately 10% of women and adolescents: more than 7.4 million women in the US and more than 176 million women worldwide. EMS is the #1 cause of disability and infertility among women in their reproductive years; 40% of infertile women have EMS. The average age at diagnosis is 28 years. Debilitating symptoms include painful menstruation, chronic pain, pain with intercourse, and infertility. In addition to human suffering, the symptom-associated productivity loss and direct health-care cost to the United States is greater than $90 billion annually. EMS occurs via retrograde menstruation, where viable endometrial tissue flows back through the fallopian tubes and into the peritoneal cavity. There it attaches to multiple foreign sites (e.g. fallopian tubes, ovarian fossa, peritoneal wall, ligaments, and bowel) and responds to hormones (Sampson, J. A., Am J Pathol 3, 93-110 143, 1927). The gold standard for the diagnosis of endometriosis has been visual inspection by laparoscopy, preferably with histological confirmation. Because there is no good noninvasive test for endometriosis, there is often a significant delay in diagnosis of this disease. Thus, there is a need for non-invasive means to detect endometriosis.

[0005] The only method to diagnosis endometriosis currently is through laparoscopic surgery with an average delay of symptom onset to diagnosis at an abysmal 4-12 years. Current efforts to use scanning technologies are experimental at best. Ultrasound is limited by field of view and is dependent on operator proficiency, while experimental MRI for endometriosis exhibits significant variability in protocols (T1T2) and utilizes gadolinium which has toxicliabilities, especially for women in reproductive age. Due to the high inconsistency of results and the low sensitivity for detecting certain pervasive subtypes of endometriosis, these methods cannot qualify as a replacement for surgery.SUMMARY

[0006] To begin bridging this considerable unmet need in endometriosis diagnosis, disclosed herein is a targeted, non-invasive diagnostic agent for endometriosis. A non-invasive diagnostic that has wide market access could provide early detection, including superficial endometriosis which can be pervasive and cause painful symptoms. For the patient, this could eliminate years of pain, worry and confusion currently experienced prior to proper diagnosis and be transformative in healthcare costs. Early intervention can also lead to better outcomes for women with endometriosis, including preserving fertility.

[0007] The development of a definitive diagnostic approach that specifically targets endometriosis lesions promises to enhance both the reliability and reproducibility of results, minimizing dependency on variables such as equipment and operator expertise. The disclosed targeted diagnostic strategy can overcome the limitations currently faced with laparoscopic assessments and contrast agents, providing a more reliable and reproducible means of diagnosing endometriosis.

[0008] The disclosed agents can be used in full body scans with traditional MRI machines that are readily available, including lower Tesla models. This includes compatibility with mobile and open MRI systems, enhancing accessibility and comfort, especially for those with claustrophobia.

[0009] The lack of precision from contrast agents like gadolinium requires the use of more powerful Telsa machines or PET / MRI scans. These machines are limited to only select imaging centers and can only at best view limited subtypes of endometriosis. The potential of our technology as targeted probes is that it could significantly improve the detection of lesions without the need for the highest-resolution equipment. Most accessible MRI machines are traditional models with lower magnetic field strengths, which are less expensive and ineffective with gadolinium contrast. The disclosed agents and methods have the potential to make endometriosis diagnosis more accessible to all populations, including underserved areas, particularly in settings where only lower-strength MRI machines are available. Further, the disclosed solution would be ready for immediate market.

[0010] The disclosed diagnostic agent can be delivered to all women suspected of having endometriosis or who are at a heightened risk for the condition, regardless of age, ethnicity, or economic status.

[0011] The imaging agent can be used with full body MRI scans to diagnose all subtypes and stages of endometriosis lesions on any organ system. The goal is to provide both the patient and the clinical provider with greater insights into disease progression and prognosis for data-driven care. For example, endometriosis on the bladder can present itself as painful UTIs while endometriosis on the appendix can present itself as excruciating appendicitis. Critically, this MRI approach can also enable the detection and characterization of malignancy, providing critical information for patients so that they can receive appropriate care.

[0012] The disclosed targeted imaging agent involves a peptide conjugated to an imaging moiety, such as an iron-based moiety. As iron / iron oxides have been proven to be safe, this solution not only shows promise to outperform but also avoid the serious side effects of gadolinium.

[0013] A key feature of an effective diagnostic is its retention in the targeted lesion alongside rapid clearance from the bloodstream, enabling a window of opportunity for imaging from blood clearance to the breakdown of the imaging agent in the lesion. Based on our research, our pharmacokinetics (PK) data in both the lesion and bloodstream suggest an imaging timeframe of at least one day. This duration is beneficial because it provides flexibility in scheduling, allowing for scans to be conducted even if there are appointment delays. Other experimental technologies based in contrast technologies have shorter imaging windows and could be impractical or difficult for clinical use.

[0014] Currently, tracking of clinical trial success for endometriosis therapies is limited to pain, which is highly subjective and non-specific. A non-surgical diagnostic method would be revolutionary and allow for the data-driven evaluation of a treatment’s impact on the extent of the disease.

[0015] The disclosed agents will not only serve as an efficacy biomarker but can also help gauge the appropriate treatment duration necessary for disease elimination. This approach could revolutionize endometriosis therapy development by eliminating the need for second look laparoscopies. Also disclosed are [3-catenin binding agents that can be used to treat endometriosis. Together, these diagnostic and therapeutic agents can be used to eliminate endometriosis as a long-term condition.

[0016] The details of one or more embodiments of the disclosure are set forth in the accompanying drawings and the description below. Other features, objects, and advantages of the disclosure will be apparent from the description and drawings, and from the claims.BRIEF DESCRIPTION OF FIGURES

[0017] FIG. 1 shows identifying subjects with endometriosis via labelled compounds 492 to 494 as imaging diagnostic. Lesion visualization is consistent between in vivo and ex vivo imaging. This is for illustration purposes and has also been performed with other compounds including compound 495.

[0018] FIG. 2 shows identifying compounds 491 and 492 biodistribution through a Cy7- labelled version of the peptide (compound 492). compound 491 is highly specific to diseased tissue, with little to no detectable amounts in other organs, including heart, lung, uterus, ovary, Gl organs, and bone. This is for illustration purposes and has also been performed with other compounds including compounds 493-495.

[0019] FIG. 3 shows vaginal administration of compound 494 which supports that vaginal administration is a viable option with the agent distributing to the lesion. This has been performed with other labelled peptides as well as other routes of administration including but not limited to SQ, IV and PO dosing.

[0020] FIG. 4 shows that peptides are permeable through vaginal tissue and that administration of peptides vaginally takes approximately 3 hours to cross the human vaginal tissue. This is consistent with the imaging performed via animal model with labelled agent (for illustrative purposes, compound 491 is shown here).

[0021] FIG. 5 shows that labelled agent / peptide can be used to determine route of elimination, which for purposes of this figure is feces for compound 492.

[0022] FIG. 6 shows spectral imaging time course for C57BI / 6 albino mice treated with compound 494. This shows that the label can be used in a living mouse to identify endometriosis lesions post blood clearance (~24h, here shown for SQ delivery) and that multiple time points can be scanned to show the time of retention of agent in endometriosis lesion, in this case from 24 to at least 48 hours post administration. This is for illustration purposes and has also been performed with other compounds including compounds 492-495 and utilizing different routes of administration which have different serum pharmacokinetic profiles.

[0023] FIG. 7 shows tracking lesion regression within the same mouse after 2 SQ doses administered weeks apart showing that daily dosing is not necessary for efficacy. This is for illustration purposes and has also been performed with other compounds including compounds (here shown 492).

[0024] FIG. 8 shows longer treatment times improve efficacy, especially at lower doses.

[0025] FIG. 9 shows the pharmacokinetic drug exposure as analyzed in serum. In comparison to FIG. 6 and FIG. 7, pharmacodynamic effects lasts longer than the pharmacokinetic drug exposure as analyzed in serum.

[0026] FIG. 10 shows a cell proliferation assay confirming that one dose of compound 491 has long pharmacodynamic effects within the lesion outlasting serum pharmacokinetics (5 days are shown post treatment). It also shows that more frequent dosing is helpful but not required for efficacy.

[0027] FIG. 11 shows compound 492 binds to target p-catenin directly within the cell, that it is cell-permeable, and it binds to target within the cytoplasm of cell and not the nucleus, and is able to be imaged without quenching of the signal by the peptide or intracellular localization of the imaging agent.

[0028] FIGs. 12A and 12B show that compound 492 binds to p-catenin with 1 hour of incubation, that it is in the cytoplasm, and that the signal is stable for at least 3 days.

[0029] FIG. 13 shows the downstream effects of the binding described in FIG. 12. FIG. 13 shows that endogenous cytoplasmic p-catenin protein starts decreasing after compound 492 P-catenin binding occurs in cytoplasm. The same trend has been observed with the nucleus fraction even though the peptide does not directly interact with the nucleus; instead sequestering excess cytoplasm p-catenin and preventing its nuclear translocation and redirecting to the membrane.

[0030] FIGs. 14A to 14C show that endogenous p-catenin protein remains stable in non- nucleus / non-cytoplasmic (membrane) fraction and that peptide binding to cytoplasmic p-catenin does not inhibit p-catenin membrane binding. FIGs. 14A to 14C shows p-catenin amount after incubation with 0.5pM compound 492 in cytoplasmic fraction (FIG. 14A). membrane fraction (FIG. 14B), or nucleus fraction (FIG. 14C).

[0031] FIG. 15 shows that imaging peptide is not detected in the healthy uterus tissue in endometriosis animals but is found in the lesion, confirming specificity to disease. This figure also shows the cellular localization of peptide which is majority at the membrane after equilibrium has been reached, which suggests Wnt4 activity of the peptides.

[0032] FIG. 16 quantitates the subcellular localization in the diseased lesion observed in FIG. 15, confirming mechanism of action observations in FIG. 15, confirming mechanism of action.

[0033] FIG. 17 shows that cell proliferation in epithelial cells of endometriosis lesions decreases after administration of compound 491 to animals, confirming mechanism of action and on target activity.

[0034] FIG. 18 shows that cell apoptosis in epithelial cells of endometriosis lesions increases after administration of compound 491 to animals, confirming mechanism of action and on target activity.

[0035] FIG. 19 shows that activated macrophages are recruited to the site of the lesion in animals after the administration of compound 491 , confirming mechanism of action and on target activity.

[0036] FIG. 20 shows that lesions extracted from animals treated with compound 491 show a decrease in RNA of 3-catenin downstream target genes, confirming mechanism of action and on target activity.

[0037] FIG. 21 shows that lesions extracted from animals treated with compound 491 show a decrease in protein of [3-catenin target genes, confirming mechanism of action and on target activity.

[0038] FIG. 22 shows the imaging of endometriosis lesions in animals which demonstrates the successful targeting and binding of a labeled peptide to 3-catenin downstream target protein MMP2.

[0039] FIG. 23 shows that the p -catenin peptide (here, compound 491 for illustrative purposes) does not alter uterine or ovarian histology. Top: As expected, in uterine cross sections, normal proliferation, glands, stromal, and normal myometrial layers were observed. Magnification = 100x. Bottom: Ovaries were normal and exhibited all stages of folliculogenesis from primordial follicles to corpus luteum. Magnification = 400x. Sections were stained with H&E.

[0040] FIG.. 24A and 24B confirms successful synthesis of MRI agent with zeta potential analysis before and after conjugation of functionalized iron oxide to compound 496. DETAILED DESCRIPTION

[0041] Unless defined otherwise, all terms of art, notations and other technical and scientific terms or terminology used herein are intended to have the same meaning as is commonly understood by one of ordinary skill in the art to which the claimed subject matter pertains. In some cases, terms with commonly understood meanings are defined herein for clarity and / or for ready reference, and the inclusion of such definitions herein should not necessarily be construed to represent a substantial difference over what is generally understood in the art.

[0042] 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 thedisclosure. Accordingly, the description of a range should be considered to have specifically disclosed all the possible subranges as well as individual numerical values within that range. For example, description of a range such as from 1 to 6 should be considered to have specifically disclosed subranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6 etc., as well as individual numbers within that range, for example, 1 , 2, 3, 4, 5, and 6. This applies regardless of the breadth of the range.Definitions

[0043] As used in the specification and claims, the singular forms “a”, “an” 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.

[0044] The terms “determining”, “measuring”, “evaluating”, “assessing, ” “assaying, ” and “analyzing” are often used interchangeably herein to refer to forms of measurement and include determining if an element is present or not (for example, detection). These terms can include quantitative, qualitative or quantitative and qualitative determinations. Assessing is alternatively relative or absolute. “Detecting the presence of” includes determining the amount of something present, as well as determining whether it is present or absent.

[0045] The terms “subject,” “individual,” or “patient” are often used interchangeably herein. A “subject” can be a biological entity containing expressed genetic materials. The biological entity can be a plant, animal, or microorganism, including, for example, bacteria, viruses, fungi, and protozoa. The subject can be tissues, cells and their progeny of a biological entity obtained in vivo or cultured in vitro. The subject can be a mammal. The mammal can be a human. The subject may be diagnosed or suspected of being at high risk for a disease. The disease can be endometriosis. In some cases, the subject is not necessarily diagnosed or suspected of being at high risk for the disease.

[0046] The term “in vivo” is used to describe an event that takes place in a subject’s body.

[0047] The term “ex vivo” is used to describe an event that takes place outside of a subject’s body. An “ex vivo” assay is not performed on a subject. Rather, it is performed upon a sample separate from a subject. An example of an “ex vivo” assay performed on a sample is an “in vitro” assay.

[0048] The term “in vitro” is used to describe an event that takes places contained in a container for holding laboratory reagent such that it is separated from the living biological source organism from which the material is obtained. In vitro assays can encompass cell-based assaysin which cells alive or dead are employed. In vitro assays can also encompass a cell-free assay in which no intact cells are employed.

[0049] As used herein, the term “about” a number refers to that number plus or minus 10% of that number. The term “about” a range refers to that range minus 10% of its lowest value and plus 10% of its greatest value.

[0050] As used herein, the terms “treatment” or “treating” are used in reference to a pharmaceutical or other intervention regimen for obtaining beneficial or desired results in the recipient. Beneficial or desired results include but are not limited to a therapeutic benefit and / or a prophylactic benefit. A therapeutic benefit may refer to eradication or amelioration of symptoms or of an underlying disorder being treated. Also, a therapeutic benefit can be achieved with the eradication or amelioration of one or more of the physiological symptoms associated with the underlying disorder such that an improvement is observed in the subject, notwithstanding that the subject may still be afflicted with the underlying disorder. A prophylactic effect 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. For prophylactic benefit, a subject at risk of developing a particular disease, or to a subject reporting one or more of the physiological symptoms of a disease may undergo treatment, even though a diagnosis of this disease may not have been made.

[0051] The terms “peptide,” “polypeptide,” and “protein” are used interchangeably herein, and refer to a polymeric form of amino acids of any length, which can include coded and non-coded amino acids, chemically or biochemically modified or derivatized amino acids, and polypeptides having modified peptide backbones.

[0052] As used herein, “peptidomimetic” means a mimetic of a peptide which includes some alteration of the normal peptide chemistry. Peptidomimetics typically enhance some property of the original peptide, such as increase stability, increased efficacy, enhanced delivery, increased half life, etc. Methods of making peptidomimetics based upon a known polypeptide sequence is described, for example, in U.S. Patent Nos. 5,631,280; 5,612,895; and 5,579,250. Use of peptidomimetics can involve the incorporation of a non-amino acid residue with non-amide linkages at a given position. One embodiment of the present disclosure is a peptidomimetic wherein the compound has a bond, a peptide backbone or an amino acid component replaced with a suitable mimic. Some non-limiting examples of unnatural amino acids which may be suitable amino acid mimics include [3-alanine, L-a-amino butyric acid, L-y- amino butyric acid, L-a-amino isobutyric acid, L-c-amino caproic acid, 7-amino heptanoic acid,L-aspartic acid, L-glutamic acid, N-c-Boc-N-a-CBZ-L-lysine, N-e-Boc-N-a-Fmoc-L-lysine, L- methionine sulfone, L-norleucine, L-norvaline, N-a-Boc-N-bCBZ-L-ornithine, N-b-Boc-N-a-CBZ- L-ornithine, Boc-p-nitro-L-phenylalanine, Boc-hydroxyproline, and Boc-L-thioproline.

[0053] The abbreviations for many of the terms used in sequences herein are defined in Table 1.

[0054] The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described.Approach

[0055] p-catenin binding peptides that can be used to treat endometriosis are disclosed in WO 2020 / 106995, which is incorporated by reference in its entirety for the teaching of these peptides and methods of making and uses thereof. As disclosed herein, these peptides can also be used for diagnosis due to their specificity for endometriosis.

[0056] In healthy cells, p-catenin is bound to E-cadherin at the cellular membrane; high concentrations of p-catenin are found at the membrane with low concentrations in the cytoplasm and nucleus73-76. Conversely, in endometriosis pathogenesis, p-catenin dissociates from the membrane by either receptor dysregulation or somatic mutation, and accumulates into the cytoplasm. Cytoplasmic p-catenin is thought to bind to estrogen receptor (ESR1) and translocate into the nucleus, p-catenin’s nuclear transcription factor activities upregulates matrix metalloproteinases (MMPs), Chloride 3 Channel (CIC-3), and other proteins that can initiate the transformation, invasion, migration, and fibrogenesis of endometriosis cells, giving rise to lesions. Epithelial to mesenchymal transition (EMT) involves TGF-p and Wnt81 , and this transformation is vital for the establishment and proliferation of endometriosis lesions by disrupting epithelial cell cytoskeleton and cell-cell organization where highly mobile mesenchymal cells are produced. Further, human endometriosis tissue exhibits high expression levels of p-catenin-activated CIC-3, matrix metalloproteinase-9 (MMP9), and SOX proteins. MMP9 supports angiogenesis, thus enabling endometrial cells to grow outside the uterus. Of note, high CIC-3 expression levels are correlated with endometriosis symptoms, including infertility, dysmenorrhea, pelvic pain, dyspareunia, and lesion diameter.

[0057] The downstream events underlying endometriosis pathogenesis all occur through P-catenin: (e.g. mutated Wnt4 [germline], mutated p-catenin [somatic], and excessive estrogen (E2) production [hormonal]). GWAS studies reveal a strong association between a WNT4 mutation and endometriosis. This is expected given the increased likelihood of endometriosis in individuals with a family history of the condition. Further, WNT4 gene expression in eutopic endometrium is significantly higher than in diseased eutopic endometrium. Importantly, the wild type WNT4 protein binds to cytoplasmic p-catenin and transports it to the membrane, thus preventing p-catenin nuclear translocation and dysregulation. The mutated or downregulated Wnt4 protein allows for p-catenin cytoplasmic accumulation, nuclear translocation, and subsequent transcription factor activity to activate IL6, MMP2, MMP7, MMP9, CIC-3, and other P-catenin-activated proteins. Our unique peptide candidates specifically target cytoplasmic p- catenin and inhibits nuclear translocation of excess cytoplasmic p-catenin by redirecting it to the membrane, similar to Wnt4 activity.

[0058] Therefore, a cytoplasmic p-catenin-specific marker is an attractive approach for targeting endometriosis, p-catenin was deemed an undruggable target due to its protein-protein interactions; however, a combination of in vitro, in vivo, and in silico tools was used to identify suitable cell permeable cyclic peptide candidates with acceptable ADME profiles for pharmaceutical use. p-catenin cytoplasmic accumulation has been observed across all stages of endometriosis, including lesions of atypical endometriosis, typical endometriosis, and cancers arising from endometriosis, make it a major target for diagnosis and treatment

[0059] Endometriosis can range from small, isolated lesions to larger areas of invasive endometriotic tissue affecting pelvic organs, including ovaries, fallopian tubes, intestines, and bladder. The American Society for Reproductive Medicine’s classification system categorizes endometriosis into 4 stages based on clinical findings of lesion characteristics (size, location, and superficial lesions or DIE) and the extent of pelvic adhesions at time of surgery. Genetic testing of lesions has confirmed all classifications and organ locations harbor somatic p-catenin mutations or mutations in genes that regulate cytoplasmic p-catenin. Further, endometriosis mesenchymal stem cells promote fibrogenesis in ovarian endometriomas through the Wnt / p- catenin pathway by paracrine production of TGF-pi and Wnt1. Multiple types of endometriotic lesions (e.g. atypical, DIE, rectovaginal, and ovarian endometrioma subtypes) can harbor oncogenic mutations in genes known to upregulate p-catenin, including ARID1A, PIK3CA, KRAS, and PPP2R1A. Together, this suggests that a cytoplasmic p-catenin-targeted diagnostic agent can address all stages and subtypes of endometriosis.

[0060] Importantly, MRI findings have been successful in the identification of malignant tumors. Combined with the targeted diagnostic agent, this approach can distinguish cancerous lesions, including those that are endometriotic. Early detection of malignant transformation is critical for proper management. Endometriosis-associated ovarian cancer (EAOC) has successfully been detected in MRI findings through: (1) identification of enhanced mural nodules, (2) growth in tumor size - specifically tumors 9 cm or larger have served as an independent predictor for ovarian cancer, and (3) an absence of T2-shading suggested to be caused by secretion from a malignant tumor that dilute hemorrhagic fluid.

[0061] The disclosed agents and methods leverage an innovative approach by using pH-sensitive peptides as pharmaceutical agents for targeted endometriosis diagnosis. Specificity to endometriosis tissue is attributed to a novel mechanism driven by pKa of weakly acidic peptides, which allows for selective permeability in diseased tissue. Further, to our knowledge, our group was the first (and only) to identify a 3-point pH differential between endometriosis and healthy cells.

[0062] Our initial toxicity findings show no observable toxicities associated with the peptides in our library. We attribute this to our unique approach of specifically targeting cytoplasmic p-catenin within endometriosis tissue via Wnt4 like activity. -catenin's membrane adherence is maintained for cellular health: disruption of [3-catenin's membrane binding can lead to proto-oncogene-like effects, whereas Wnt4 activity is essential for uterine decidualization, Gl tract maintenance and preventing anemia. Additionally, our approach is nonhormonal.

[0063] The disclosed peptides have shown to be both potent and exhibit high safety profiles, even over 2000 times efficacious exposure level. In addition, peptide degradation products pose little to no risk for toxicities (unlike small molecules). MRI-compatible, iron- conjugated peptides are therefore expected to be exceptionally safe, as iron moieties are not associated with the toxicities commonly linked to gadolinium.

[0064] The use of targeted molecular agents for diagnosing endometriosis significantly enhances precision by binding exclusively to diseased cells, outperforming traditional methods that depend on non-specific contrast absorption. The disclosed methods involves highly specific peptides designed for pinpoint accuracy in detecting endometriosis lesions down to submillimeter levels. This approach is expected to improve reproducibility, reliability, sensitivity and specificity across all lesion subtypes and organs using standard MRI equipment, thereby enhancing accessibility, and reducing the costs of scans.P-catenin-Binding Peptides

[0065] Disclosed herein are [3-catenin-binding agents that can be useful in a variety of imaging, diagnostic and / or therapeutic settings including, for example, detecting, treating, inhibiting, preventing, monitoring, staging or reducing endometriosis (EMS) and its related symptoms.

[0066] In some embodiments, these agents can be useful in imaging, diagnostic, and / or therapeutic setting for other diseases associated with dysregulated Wnt / p-catenin pathway include colorectal cancer, hepatocellular carcinoma, endometrial cancer, ovarian cancer, medulloblastoma, glioblastoma, melanoma, and Alzheimer's disease.

[0067] In some embodiments, these agents can be utilized for diagnostic, imaging, and treatment of other gynecological conditions include endometriosis, uterine fibroids, adenomyosis, ovarian cysts, polycystic ovary syndrome (PCOS), endosalpingiosis and cervical, uterine, and vaginal polyps, and / or its symptoms.

[0068] In some embodiments, the [3-catenin binding agent comprises a peptide. p- catenin-binding peptides are described in WO 2020 / 106995, which is incorporated by reference in its entirety for the teaching of these peptides and methods of making and uses thereof.

[0069] As disclosed herein, the disclosed p-catenin-binding peptides can be used to selectively target an agent to endometriosis lesions. In some embodiments, the peptides target diseased tissue with pathogenesis relating to [3-catenin dysregulation. In some embodiments, the agent and peptide are able to identify diseased tissue. In some embodiments, the agent and peptides are preferential for diseased tissue over healthy tissue. In some embodiments, the peptides are not detected in healthy tissue, such as uterine tissue. Without wishing to be bound by theory, the peptides may be selective for [3-catenin having a conformation of being cell permeable and destined for the cytoplasm where there is an excess of target being available in the diseased tissue (cytoplasmic [3-catenin).

[0070] The disclosed peptides may also be able to directly visualize and / or address EMS etiology, reverse disease progression, and prevent or delay EMS recurrence. In some cases, the [3-catenin-binding peptides bind specifically to EMS lesions. In some cases, it can shrink existing EMS lesions. In some cases, [3-catenin-binding peptides can inhibit dysregulation of the Wnt pathway, which is believed to be one of the underlying causes of EMS. The [3-catenin binding peptides can sometimes limit EMS pathogenesis by preventing endometrial cell invasiveness. The [3-catenin binding peptides can sometimes limit EMS pathogenesis by inducing apoptosis. The [3-catenin binding peptides can sometimes induce macrophages which help clear lesions. Alternatively or in addition, the [3-catenin-binding peptides can suppress epithelial to mesenchymal transition (EMT). This suppression can limit proliferation of endometriotic lesions. Alternatively or in addition, the [3-catenin-binding peptides can sometimes reduce paracrine production of TGF-[31 and Wnt1 , which in turn can diminish fibrogenesis in ovarian endometriomas. In some cases, the [3-catenin binding peptides reduces disease burden and inflammation caused by the disease. In some cases, downregulating cytoplasmic [3- catenin can reduce expression of procytokine IL-6 by inhibiting the [3-catenin complex with TCF / LEF transcription factors, and thus expression of target genes, including but not limited to IL-6.

[0071] In some cases, [3-catenin binding peptides bind to cytoplasmic [3-catenin and shuttle back to membrane in Wnt4 like activity. In some cases, Wnt4 like activity / rescue mechanism can help maintain bone and colon regeneration while downregulating integrity. In some cases, peptide agents displaying Wnt4 activity could help with recurrent implantation failure. In some cases, the Wnt4 rescue mechanism can result in a downstream cytoplasmic [3- catenin-specific canonical Wnt antagonism through redirecting [3-catenin’s nuclear localization to the membrane, which can inhibit or modulate aberrant transcription factor activity while maintaining key cell-cell junctions. These is an attractive approach for imaging, diagnosing, andtreating diseases driven by misregulated p-catenin, including but not limited to endometriosis, adenomyosis, fibroids, colon cancer, skin cancer, endometrial cancer, and ovarian cancer.

[0072] Moreover, the [3-catenin-binding peptides can sometimes reduce expression of downstream proteins, including but not limited to Matrix Metalloproteinase 9 (MMP9), Matrix Metalloproteinase 2 (MMP2), Matrix Metalloproteinase 7 (MMP7), SOXs (SRY-related high- mobility group box transcription factors), lnterleukin-6 (IL6), Fibronectin 1 (Fn1), or Chloride C3 Channel (CIC-3). In some embodiments, it can result in reduced angiogenesis. Inhibiting or reducing angiogenesis can inhibit the ability of endometrial cells to grow outside the uterus.

[0073] The peptides described herein may be optimized for affinity for their desired target, absorption or cellular uptake, stability, protease resistance, and other factors. The affinity of a peptide for its intended target can be altered using a variety of methods. For example, the affinity can be improved with amino acid substitutions. Additionally, modifying the size of a peptide, modifying the position of cyclization, increasing hydrophobicity, and modifying the amino acids or incorporating artificial amino acids into the structure of the peptide can affect the affinity of a peptide. Exemplary modifications include incorporating N-methyl and alpha-methyl amino acids.

[0074] Among the provided embodiments are peptide, peptidomimetic, and / or p-catenin binding agents comprising the same or similar structures and properties as the molecules described herein. Such peptidomimetic agents include small protein-like chains designed to mimic a peptide. These include peptidomimetic agents comprising modifications of a peptide, or by designing similar systems that mimic peptides, such as peptoids and [3-peptides.

[0075] Therefore, disclosed herein are peptides and peptidomimetic agents comprising an amino acid sequence having the formula X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12- X13, wherein: Xi 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 with between 4 and 12 carbons; X5is M, Norleucine, Orn, D, E, K, H, R, K, 8-aminooctanoic acid, an amino carboxylic acid with between 4 and 12 carbons or null; X6is W, Y, F, or N-methyl A; X7is F, I, L, Chg, Cha, or Tie; X8is L, I, or A; X9 is L, I, or A; X10 is C, S, A, Abu, C(me), or S(Bzl); Xn is F, H, A, K, E, Chg, Cng, or Orn; X12 is W, Y, A, or F; and X13 is G, GABA, or null; X14 is V, I, L, norvaline or null; X15 is W, Y, F, or null; and X16 is R, G, or null.

[0076] Provided herein are peptides and peptidomimetic agents comprising an amino acid sequence having the formula X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12- X13-X14-X15-X16, wherein: R is NH2, acetylation, stearic acid, palmitic acid, myristic acid, lauric acid, a Ci-Cs hydrocarbon, a Ci-Cs fatty acid, or null; Xi is M, G, beta alanine, norleucine, norvaline, or null;X? 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; X6is I, Chg, H, or L; X7is L or I; Xs is D, N, E, or Q; X9is D, E, K, Q, or Orn; X10 is H or methyl-H; Xu is V, alpha methyl V, Chg, L I, or norvaline; X12 is Q, Aib, S, R, or N; X13 is R, K, citrulline, Orn, D, or E; X14 is V, I, L, or norvaline; X15 is W, Y, or F; and X16 is R, G, or null.

[0077] In some embodiments, the peptide or peptidomimetic has the amino acid sequence Ac-CW-{NMeA}-Q-K*ILDD*-H-{Chg}-QRVWG-NH2(SEQ ID NO:496) or (Ahx)-NH-W- NMeA-QK*ILDD*H-Chg-QRVWG-NH2(SEQ ID NO:497).

[0078] Therefore, disclosed herein are peptides and peptidomimetic agents comprising an amino acid sequence of any one of SEQ ID NO:1-SEQ ID NO:499, or peptides and peptidomimetic agents 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-SEQ ID NO:499.

[0079] In some embodiments, the imaging moiety is an iron oxide nanoparticle.Therefore, in some embodiments, the imaging agent has the formula:

[0080] In some embodiments the disclosed peptide or peptidomimetic can be covalently conjugated to superparamagnetic iron oxide nanoparticles (SPIONs) (e.g. PEGylated SPIONs)via a linker using the method described in Liu et al. Nanoscale 2012 4:2306, which is incorporated by reference in its entirety for the teaching of this method.

[0081] In some embodiments, the imaging moiety is iron chelated to 1 ,4,7,10- tetraazacyclododecane-1 ,4,7,10-tetraacetic acid (DOTA). Therefore, in some embodiments, the imaging agent has the formula {DOTA-[lron]}-(Ahx)-NH-W-NMeA-QK*ILDD*H-Chg-QRVWG- NH2(Compound 499, SEQ ID NO:499).

[0082] In some embodiments, the peptide has a pKa value from about 4.5 to about 7.2, including m about 4.5 to about 6, from about 5.5 to about 7.0, from about 5 to about 7.2, and from about 4.5 to about 7.0.

[0083] In some embodiments, the peptide binds directly to [3-catenin. In some embodiments, the binding occurs in the cytoplasm. In some embodiments, the peptide is an inhibitor of p-catenin translocation to the nucleus. In some embodiments, the peptide allows for binding of p-catenin to the membrane. In some embodiments, the peptide acts like a Wnt4 rescue. In some embodiments, the peptide does not decrease membrane binding of p-catenin. In some embodiments, the peptide prevents p-catenin acting as a transcription factor to oncogenes, MMP2, MMP7, SOXs, IL6, Fn1 or Chloride C3 Channel (CIC-3). In some embodiments, the peptide prevents transformation, invasion, migration, fibrogenesis, or any combination thereof, of endometriosis (EMS) cells. In some embodiments, the peptide activates cell apoptosis and macrophages in diseased tissue. In some embodiments, generalized necrosis is not occurring. In some embodiments, the peptide does not affect HPX axis (i.e. , hormonal cyclicity). In some embodiments, the peptide act like Wnt4 agonists. In some embodiments, the peptide does not decrease p-catenin E-cadherin binding. In some embodiments, the peptide prevents oncogenic transcription factor activity. In some embodiments, the peptide downregulates Wnt pathway activity with an ECsoof less than or less than about 50 pM, about 30 pM, about 10 pM, about 5 pM, about 1 pM, 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 non-naturally occurring. In some embodiments, the peptide is a circularized peptide. In some embodiments, the peptide is a bicyclic peptide. In some embodiments, the peptide is circularized with a Cys-Cys disulfide bond. In some embodiments, the peptide is circularized with an amide bond. In some embodiments, the amide bond is head- to-tail between N-terminus and C-terminus. In some embodiments, the amide bond is head-to- side chain between N-terminus and an internal COOH. In some embodiments, the amide bondis side chain-to-tail between an internal NH2and C-terminus. In some embodiments, the amide bond is side chain-to-side chain between an internal NH2and an internal COOH. In some embodiments, the peptide is circularized using hydrocarbon stapling. In some embodiments, the peptide is circularized 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, less than 44, less than 45, less than 46, 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 amino acid residues. In some embodiments, the peptide comprises one or more non-natural amino acids. In some embodiments, the one or more nonnatural amino acids are N-methyl amino acids.

[0084] Exemplary peptide sequences include compounds 1-499 (SEQ ID NO: 1-SEQ IDNO: 499), as listed in Table 2.

[0085] Proteins downstream from [3-catenin can also be targeted for endometriosis and other gynecological diseases for treatment and imagining. Labelled peptide targeting MMP2 shows that lesions are able to be visualized (e.g., see FIG. 13). These agents can also be used for other diseases that are affected by [3-catenin dysregulation.Peptide Synthesis

[0086] The peptides described herein can be produced by synthetic methods. For example, Solid-Phase Peptide Synthesis (SPPS) allows the rapid assembly of a peptide chain through successive reactions of amino acid derivatives on an insoluble porous support. The solid support often includes small, polymeric resin beads functionalized with reactive groups (such as amine or hydroxyl groups) that link to the nascent peptide chain. Since the peptide remains covalently attached to the support throughout the synthesis, excess reagents and side products can be removed by washing and filtration. Each amino acid to be coupled to the peptide chain N-terminus can be protected on its N-terminus and side chain using appropriate protecting groups such as Boc (acid-labile) or Fmoc (base-labile), depending on the side chain and the protection strategy used (see below).

[0087] The general SPPS procedure is usually one of repeated cycles of alternate N- terminal deprotection and coupling reactions. The resin can be washed between each step. First, an amino acid is often coupled to the resin. Subsequently, the amine is usually deprotected, and then coupled with the free acid of the second amino acid. This cycle can repeat until the desired sequence has been synthesized. SPPS cycles may also include capping steps that can block the ends of unreacted amino acids from reacting. At the end of the synthesis, the crude peptide can be cleaved from the solid support. This step can often include simultaneously removing all protecting groups using a strong acid like trifluoroacetic acid or a nucleophile. The crude peptide can be precipitated from a non-polar solvent like diethyl ether in order to remove organic soluble by products. The crude peptide can be purified using reversed- phase HPLC. Byproducts may be removed using continuous chromatography processes such as MCSGP to maximize the yield without sacrificing on purity levels.

[0088] The peptides described herein can be cyclic peptides. Cyclic peptides can offer several advantages in certain circumstances. For example, cyclic peptides often metabolizeslower due to their higher resistance toward proteases than non-cyclic counterparts; on the other hand, they have longer-acting depot effect than their corresponding linear counterparts. They can be used to mimic the structure of biologically active peptides (e.g., peptide hormones) and are able to bind drug targets in vivo.

[0089] The peptides described herein can have some flexibility in the rings. Flexible rings allow for different confirmations and allow for flexibility to bind to the target which target undergoes different confirmational structures. Peptides are able to penetrate cells and can exert their effect on intracellular protein targets (cell permeable peptides). The large surface area of the cell permeable peptides gives them advantages over small molecules in their ability to disrupt specific signaling pathways by inhibiting targeted protein-protein interactions. This occurs when peptides bind to p-catenin, it allows for binding to occur as p-catenin changes its structure from cytoplasmic (stabilized) to destabilized structures that allow for membrane binding. The ability of flexible peptides to bind to cytoplasmic p-catenin allows for peptides to show activity and efficacy, while the ability of the same p-catenin bound peptide to go to the membrane of the cell allows for safety during use. Therefore, flexible peptides are advantageous for diagnostic and imaging use can act as Wnt4 agonist because they are flexible and allow for both safety and efficacy. Peptides have a conformational structure that can mimic the molecular structures that are typically found at the interface of protein-protein interactions.

[0090] The peptides described herein can by cyclized using a disulfide bridge. Peptide disulfide bridge can link two thiol (SH) groups from the side chain of cysteines or cysteine analogues. Undesired linkage can be prevented by using appropriate protecting group chemistry to effect either specific intra- or intermolecular oxidation. In general, a disulfide bridge can be formed as follows: intermolecular (two peptide molecules are linked via the disulfide bridge), resulting in either: homodimers (two identical peptides) or heterodimers (two different peptides), or intramolecular (cyclization within one peptide molecule).

[0091] Cyclic peptides can also be synthesized by linking the amino (N) terminus of the 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 can also be used to construct cyclic peptides via an amide bond. Depending on functional groups of a peptide, cyclic peptide synthesis often uses one of four different methods: head-to-tail between N-terminus and C-terminus; head-to-side chain between N-terminus and an internal COOH (e.g. the p-COOH-group of Asp or y-COOH- group of Glu); side chain-to-tail between internal NH2s and C-terminus (e.g. the e-NH2-group of Lys); and side-chain-to-side-chain between an internal NH2 and an internal COOH (e.g. the s- NH2-group of Lys with either the p-COOH-group of Asp or y-COOH-group of Glu).

[0092] The peptides described herein can be cyclized using stapled peptide synthesis. Peptide stapling is typically achieved by incorporating a, a disubstituted non-natural amino acids bearing terminal olefin tethers of varying length. Subsequent olefin metathesis creates carboncarbon bond tethers between amino acid side chains to cyclize the peptide. Alternatively, stapled peptides can be generated using Fmoc solid-phase synthesis chemistry as is generally known in the art.

[0093] The peptides described herein can also by cyclized using click chemistry. Clickable functional groups can be incorporated into peptides at time of synthesis using different combinations of protected amino acids modified with an alkyne group, followed by click reaction with an azido-acid. The resulting peptides are detached off the resin to give triazole-containing peptides. These functional groups can also be introduced via post synthesis modification to produce structurally constrained peptides.

[0094] The peptides described herein can have modifications to increase stability. It has been generally shown that N-methyl modifications can increase 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 by reference herein for such disclosure). In some embodiments described herein, peptides are modified with one or more N-methyl analogues to natural amino acids. Incorporation of one or more N-methyl amino acids can increase proteolytic resistance form 10-fold to 10000-fold over resistance of peptide consisting of natural residues. In some embodiments, the proteolytic resistance of peptides containing one or more N-methyl amino acids is at least 10-fold, at least 20-fold, at least 30-fold, at least 40-fold, at least 50-fold, at least 60-fold, at least 70-fold, at least 80-fold, at least 90-fold, at least 100-fold, at least 200- fold, at least 400-fold, at least 500-fold, at least 600- fold, at least 700-fold, at least 800-fold, at least 900-fold, at least 1000-fold, at least 2000-fold, at least 3000-fold, at least 4000-fold, at least 5000-fold, at least 6000-fold, at least 7000-fold, at least 8000-fold, at least 9000-fold, at least 10,000-fold over resistance of the peptide consisting of natural residues.Imaging Agent and Methods

[0095] Disclosed herein are imaging agents that involve one or more of the disclosed - catenin-binding peptides conjugated to an imaging moiety. Also disclosed are methods of detecting diseased tissue in a subject that involve administering to the subject a disclosed imaging agents and imaging the subject or object for the presence of the imaging moiety in the subject’s body.

[0096] In some embodiments, the diseased tissue is endometriosis lesions. In some embodiments, the diseased tissue are fibroids. In some embodiments, the diseased tissue is colon, ovarian, endometrial, and skin cancer tumors.

[0097] Herein are describing methods in all living cells and species. In some embodiments, subjects are humans. In some embodiments subjects are animals. In some embodiments, subjects are cultured cells. In some embodiments subjects are ex vivo diseased tissue removed from animals. In some embodiments subjects are ex vivo diseased tissue removed from humans.

[0098] In some embodiments, the imaging moieties are intrinsically cell permeable. In other embodiments, the imaging moieties are not inherently cell permeable but become permeable when conjugated to a cell permeable peptide, effectively acting as cargo to the cell permeable peptide.

[0099] The disclosed peptides and peptidomimetics can be labeled with imaging moieties in order to serve as in vivo imaging probes. In some embodiments, the label is in the N-terminus. In some embodiments, the label is in the C-terminus. In some embodiments, the label is attached to an amino acid sidechain. In some embodiments the label substitutes atoms or molecules within the amino acid sequence (i.e. , radiolabel). In some embodiments, the label is followed by a modification. In some embodiments the modification is lipid, acetylation, and / or amidation.

[0100] The advantage of using the imaging agents described herein is that the peptides are bindings to targets in the diseased tissue, and thus helping with signal in the diseased tissue to background noise of healthy tissue. This means that the agents can help visualize very small lesions (<1mm as described in examples in mice) and / or lesions that are deep in the body whereas the limit of detection for scanning technologies without imaging agents are much higher or non-specific. This is critical in detecting a metastatic disease like endometriosis or cancer where the size and spread of the diseased tissue is unknown in a patient. Ensuring that all lesions or tumors are identified is critical for pathology, staging, and recurrence.

[0101] These imaging agents can revolutionize medical imaging, disease identification, and staging. In some embodiments, the technology may be used for initial diagnosis, providing high-resolution, comprehensive imaging to facilitate accurate disease detection. Other embodiments may use the tool for surgical preparation, assisting in the detailed planning of procedures by offering in-depth visualization of the disease site. Furthermore, the diagnostic can serve as a basis for tracking the ongoing progression of a disease, helping monitor patient health over time and adjust treatment protocols accordingly. There are embodiments where thetechnology may assess the effectiveness of therapies during and after a treatment course, providing valuable feedback on treatment efficacy. Repeat scanning could be performed in some embodiments, which is particularly useful for chronic or long-term conditions that require continuous monitoring. In some embodiments, the imaging agent is a companion diagnostic for the therapeutic. In some embodiments, the diagnostic tool can be combined with therapeutic techniques (i.e. , theranostics) where diagnostics and therapeutics are integrated into a single system for a more holistic and personalized approach to patient care. In some embodiments, the diagnostic compound can also elicit therapeutic efficacy or effects. In some embodiments, the peptide imaging agent also can be used to assess properties of the active pharmaceutical ingredient (API) or drug product including but not limited to bioavailability, PK, and route of elimination.

[0102] Some widely used imaging moieties include organic dyes for optical imaging, radionuclides for PET and SPECT imaging and Gd3+ chelators for MR imaging. The functional groups of peptides available for conjugation include but are not limited to the e-amino group on lysine side chains, the guanidinium group on arginine side chains, the carboxyl groups on aspartic acid or glutamic acid, the cysteine thiol, and the phenol on tyrosine. The most common conjugation reactions are carbodiimide / N-hydroxysuccinimidyl (EDC / NHS) mediated carboxyl and amine coupling, maleimide conjugation to thiol groups, and diazonium modification of the phenol on tyrosine.

[0103] Radiolabeled peptides can also be used as peptide-based imaging agents. Radionuclide based PET / SPECT imaging could detect the imaging agents at micromolar to picomolar concentration. Therefore, it is possible to minimize the usage of peptides in order to reduce any adverse biological effect. To develop peptide based probes for PET / SPECT imaging, radionuclides should be labeled onto targeting peptides. Several radionuclides employed for peptide labeling are99mTc,123l, and 1111n for SPECT imaging and18F,64Cu and68Ga for PET imaging. Generally, these radionuclides are attached to the peptides via chelators or post-synthetic radiolabeling groups. Some widely-used chelators include 1 ,4,8,11- tetraazacyclotetradecane-1 ,4,8,11-tetraacetic acid (TETA), 1 ,4,7,10-tetraazacyclododecane- 1 ,4,7,10-tetraacetic acid (DOTA), 1 ,4,7-triazacyclononane-1,4,7-triacetic acid (NOTA), NODAGA (1 ,4,7-triazacyclononane,1 -glutaric acid-4, 7-acetic acid), diethylenetriaminepentaacetic acid (DTPA), mercaptoacetylglycilgylcilgylicine (MAG3), diaminedithiols (DADT), 2-hydrazidonicotinic acid (HYNIC), N-succinimidyl-4-18F- fluorobenzoate (18FSFB), N-succinimidyl-5-iodo-3-pyridinecarboxylate (SIB), and N- succinimidyl-3-iodobenzoate (SI PC).

[0104] Herein, the composition of peptides described in this patent can be labelled with contrast agents for CT scans, including but not limited to iron-based, iodine-based, or goldbased contrast agents or other types of imaging moieties. These agents, containing high atomic number elements such as iodine or gold, exhibit exceptional X-ray attenuation properties, thereby enabling improved visualization during CT scans.

[0105] The design of fluorophore labeled peptides is similar to radiolabeled peptides except that fluorophores are used to replace radionuclides. Various dyes are commercially available (for examples, Cyanine dyes from GE Healthcare and Alexa Fluor dyes from Invitrogen). Although the fluorescence imaging has the advantages of high resolution, non- invasive and safe detection, the use of fluorophores in vivo is limited by the light penetration and tissue autofluorescence. The introduction of hybrid derivatives containing both a fluorescent tag and a radioactive label opens a new avenue to the field of image-guided surgery.

[0106] In some embodiments, the fluorescent agent is selected from the group consisting of a fluorophore, fluorochrome, dye, pigment, fluorescent transition metal, and fluorescent protein. In some embodiments, the fluorescent agent is selected from the group consisting of Cy5, Cy5.5, Cy2, FITC, TRITC, Cy7, FAM, Cy3, Cy3.5, Texas Red, ROX, HEX, JA133, AlexaFluor 488, AlexaFluor 546, AlexaFluor 633, AlexaFluor 555, AlexaFluor 647, DAPI, TMR, R6G, GFP, enhanced GFP, CFP, ECFP, YFP, Citrin, Venus, YPet, CyPet, AMCA, Spectrum Green, Spectrum Orange, Spectrum Aqua, Lissamine and Europium.

[0107] Such peptides may include but are not limited to SEQ ID NO. 491 -SEQ ID NO. 499. Compound 495 variant contains a mini-PEG (polyethylene glycol) linker inserted between the Cy7-labelled cysteine and the subsequent amino acid. This modification might be beneficial for solubility, stability, or biological availability.

[0108] The peptides described here can also be optimized into dimeric or multimeric forms to change receptor targeting capability. Dimeric and tetrameric forms of analogous peptides have the potential to exhibit higher binding affinities compared to their monomeric counterparts. To optimize these multimeric peptide complexes, different types of linkers can be introduced between the peptides and imaging moieties. These linkers, which vary in length, flexibility, and hydrophilicity, can significantly influence the overall binding affinity of the peptides. Using this strategy, heterodimers can be constructed with the use of one or more peptides described herein that could enable the targeting of multiple over targets, potentially leading to enhanced imaging outcomes.

[0109] In certain embodiments, fluorescent agent(s) have excitation and emission wavelengths in the red and near infrared spectrum in the range. In certain embodiments,fluorescent agent(s) have excitation and emission wavelengths ranging from 400 to 1300 nm, or from 440 to 1100 nm, or from 550 to 800 nm, or from 600 to 900 nm. Use of this portion of the electromagnetic spectrum maximizes tissue penetration and minimizes absorption by physiologically abundant absorbers such as hemoglobin (<650 nm) and water (>1200 nm). Probe species with excitation and emission wavelengths in other spectrums, such as the visible and ultraviolet light spectrum, can also be employed in certain embodiments. In particular, fluorophores such as certain carbocyanine or polymethine fluorescent fluorochromes or dyes can be used as the fluorescent agent, e.g., U.S. Pat. No. 6,747,159 to Caputo et al. (2004); U.S. Pat. No. 6,448,008 to Caputo et al. (2002); U.S. Pat. No. 6,136,612 to Della Ciana et al. (2000); U.S. Pat. No. 4,981 ,977 to Southwick, et al. (1991); 5,268,486 to Waggoner et al. (1993); U.S. Pat. No. 5,569,587 to Waggoner (1996); U.S. Pat. No. 5,569,766 to Waggoner et al. (1996); U.S. Pat. No. 5,486,616 to Waggoner et al. (1996); U.S. Pat. No. 5,627,027 to Waggoner (1997); U.S. Pat. No. 5,808,044 to Brush, et al. (1998); U.S. Pat. No. 5,877,310 to Reddington, et al. (1999); U.S. Pat. No. 6,002,003 to Shen, et al. (1999); U.S. Pat. No. 6,004,536 to Leung et al. (1999); U.S. Pat. No. 6,008,373 to Waggoner, et al. (1999); U.S. Pat. No. 6,043,025 to Minden, et al. (2000); U.S. Pat. No. 6,127,134 to Minden, et al. (2000); U.S. Pat. No. 6,130,094 to Waggoner, et al. (2000); U.S. Pat. No. 6,133,445 to Waggoner, et al. (2000); U.S. Pat. No. 7,445,767 to Licha, et al. (2008); U.S. Pat. No. 6,534,041 to Licha et al. (2003); U.S. Pat. No. 7,547,721 to Miwa et al. (2009); U.S. Pat. No. 7,488,468 to Miwa et al. (2009); U.S. Pat. No. 7,473,415 to Kawakami et al. (2003); also WO 96 / 17628, EP 0 796 111 B1 , EP 1 181 940 B1 , EP 0 988 060 B1 , WO 98 / 47538, WO 00 / 16810, EP 1 113 822 B1 , WO 01 / 43781 , EP 1 237 583 A1 , WO 03 / 074091 , EP 1 480 683 B1 , WO 06 / 072580, EP 1 833 513 A1 , EP 1 679 082 A1 , WO 97 / 40104, WO 99 / 51702, WO 01 / 21624, and EP 1 065 250 A1 ; and Tetrahedron Letters 41 , 9185-88 (2000).

[0110] Exemplary fluorescent agents include, for example, the following: Cy5.5, Cy5, Cy7.5 and Cy7 (GE® Healthcare); AlexaFluor660, AlexaFluor680, AlexaFluor790, and AlexaFluor750 (Invitrogen); VivoTag™680, VivoTag™-S680, VivoTag™-S750 (VisEn Medical); Dy677, Dy682, Dy752 and Dy780 (Dyomics®); DyLight® 547, and / or DyLight® 647 (Pierce); HiLyte Fluor™ 647, HiLyte Fluor™ 680, and HiLyte Fluor™ 750 (AnaSpec®); IRDye® 800CW, IRDye® 800RS, and IRDye® 700DX (Li-Cor®); ADS780WS, ADS830WS, and ADS832WS (American Dye Source); XenoLight CF™ 680, XenoLight CF™ 750, XenoLight CF™ 770, and XenoLight DiR (Caliper® Life Sciences); and Kodak® X-SIGHT® 650, Kodak® X-SIGHT 691 , Kodak® X-SIGHT 751 (Carestream® Health).

[0111] In some embodiments, the imaging moiety is an MRI agent. In some embodiments, the MRI agent can be Gd(-salts), Ir(-salts), functionalized iron oxide, iron oxides, paramagnetic chemical exchange saturation transfer (CEST) agents, 19F active materials, manganese, melanin, or a substance that shortens or elongates T1 or T2 and a combination thereof. In certain embodiments, a Gd MRI agent can be a compound such as DOTA-Gd, DTPA-Gd, Gd within a polymeric chelator, and Gd immobilized by negative charges on a layer. In certain embodiments, as described, an Iron MRI agent can be a compound such as DOTA-lr, DTPA-lr, functionalized Iron oxide, functionalized iron oxide within a polymeric chelator, and Iron immobilized by negative charges on a layer.

[0112] In certain embodiments, an iron oxide MRI agent can be a compound such as a small paramagnetic iron oxide (SPIO) or an ultrasmall SPIO with or without a dextran or other stabilizing layer or superparamagnetic iron oxide nanoparticles (SPION). Superparamagnetic iron oxide nanoparticles (SPIONs) themselves are not inherently cell permeable. However, their cellular uptake can be improved through surface functionalization with appropriate ligands or peptides that facilitate internalization by target cells. This can be achieved through various mechanisms, such as receptor-mediated endocytosis, macropinocytosis, or direct membrane penetration.

[0113] In some embodiments, functionalizing the imaging agents with cell-penetrating peptides (CPPs) can enhance their cellular uptake. CPPs are a class of short peptides that can efficiently translocate across cell membranes and facilitate the internalization of cargo molecules, such as nanoparticles, proteins, or nucleic acids. By attaching CPPs to SPIONs or other imaging agents, the cell permeability of the nanoparticles can be improved, enabling their entry into target cells for various biomedical applications, including but not limited to imaging diagnostics for MRI. In certain embodiments, a paramagnetic CEST MRI agent can be a compound such as lanthanide complexes.

[0114] In some embodiments, MRI agents can be linked to a layer via a linkage such as a maleimide linkage, NHS ester, click chemistry, or another covalent or non-covalent approach or a combination thereof. In some embodiments, MRI agents can also be loaded without addition of any exogenous agent, i.e. , only layer(s) and MRI agent.

[0115] In some embodiments, the imaging moiety is an agent for use in PET, which may include, but is not limited to18F,64Cu,11C,13N,150,68Ga,89Zr,82Rb,14C,26AI,124l,76Br, and52Fe. In some embodiments, the imaging moiety is an agent for use in SPECT, which may include, but is not limited to99mTc,67Ga,192lr,111In,123l,201TI,131l,125l,133Xe, and57Co. In some embodiments, the imaging moiety is an alpha-emitting radionuclide, which may include, but isnot limited to At-211, Bi-212, Bi-213, Ra-223, Ac-225, Th-227, and Po-210. In some embodiments, the imaging moiety is a beta-emitting radionuclide, which may include, but is not limited to Cu-67, Y-90, Ag-111 , 1-131 , Pm-149, Sm-153, Ho-166, Lu-177, Re-186, Re-188, Sr- 89, P-32, and TI-204.

[0116] In some embodiments, the imaging moiety is an ultrasound contrast agent, such as a microbubble. This can improve the specificity of ultrasound imaging and enable targeted delivery of drugs or other imaging, diagnostic, or therapeutic agents to endometriosis lesions. There are several types of microbubbles, including sulfur hexafluoride-filled microbubble contrast agents, perfluorocarbon-filled microbubbles, and nitrogen-filled microbubbles. These agents typically have a shell composed of lipids, proteins, or polymers, and an average diameter of 1-10 micrometers, allowing them to pass through capillaries and circulate within the bloodstream. One commercially available example of a microbubble ultrasound contrast agent is SonoVue® (also known as Lumason® in the United States), which is a sulfur hexafluoride- filled microbubble contrast agent with a phospholipid shell. SonoVue® enhances the visualization of blood vessels and tissue perfusion in various applications, such as echocardiography and focal liver lesion characterization.

[0117] A microbubble shell can be functionalized with a peptide peptidomimetic disclosed herein. This process typically involves conjugating the peptide to the lipids, proteins, or polymers that form the microbubble shell.

[0118] Microbubbles themselves are generally too large to enter cells directly. However, by combining microbubbles with ultrasound energy, it is possible to enhance cell membrane permeability and facilitate the transport of molecules into cells via sonoporation. By applying ultrasound energy to the target area, the microbubbles to oscillate or collapse, generating mechanical forces that transiently increase the permeability of nearby cell membranes. This process, sonoporation, allows the attached molecules (e.g., drugs, genes) to enter the cells. By monitoring the treatment progress using ultrasound imaging. The microbubbles can enhance ultrasound image contrast, allowing for real-time visualization and assessment of the target area.

[0119] In some embodiments, provided particles are imaged (or susceptible to imaging) by an imaging modality selected from the group consisting of MRI, PET, SPECT, CT, X-ray, ultrasound, photoacoustic detection, fluorescent and / or Raman spectroscopy, and / or combinations thereof.

[0120] In some embodiments, the imaging agent is given during the scan. In some embodiments, the image is given at less than 15 min, less than 30 min, less than 60 min, lessthan 90 min, less than 2 hours, less than 3 hours, less than 4 hours, less than 5 hours, less than 6 hours, less than 7 hours, less than 8 hours, less than 9 hours, less than 10 hours, less than 11 hours, less than 12 hours, less than 1 day, less than 2 days, less than 3 days, less than 4 days, less than 5 days, less than 6 days, less than 7 days, less than 2 weeks, less than 3 weeks, or less than 1 month before the scan. In some embodiments, the scan may help identify diseased tissue. In some embodiments, the scan may be read as a contrast. In some embodiments, the signal remains in the lesion for a period of time beyond the PK of the drug in the system.

[0121] In some embodiments, the signal can be used to characterize properties of the API and / or drug product. In some embodiments, signal can be used to track metabolites. In some embodiments, signal can be used to determine the release profile at the site of application. In some embodiments, signal can be used to track ADME profile (i.e., bioavailability or root of elimination) of drug.Linkers

[0122] In some embodiments, peptides may be conjugated with linkers. Linker moieties can be used to covalently link one or more fluorophores, quenchers, biological modifiers and non-fluorescent reporters to an enzymatically cleavable oligopeptide or to an optional biological modifier to produce agents of the present invention. It is understood that there is no particular structural, size or content limitation of a linker, if present. Linkers can include, for example, a variety of functional groups such as maleimide, dithiopyridyl, thiol, azide, alkene, or alkyne that permit the assembly of molecules of diverse architecture.

[0123] Linkers can be homofunctional linkers or heterofunctional linkers. For example, amine (NH2)-functionalized moieties can be reacted with bifunctional cross-linkers designed to react with amino groups. Particularly useful conjugation reagents that can facilitate formation of a linker or facilitate covalent linkage between, for example, a fluorophore, and an enzymatically cleavable oligopeptide can include a N-hydroxysuccinimide (NHS) ester and / or a maleimide. The NHS ester can react with the amine group of, for example, a peptide or fluorophore. The maleimide can react with the sulfhydryl group of another molecule. Other particularly useful linker moieties are bifunctional crosslinkers such as N-succinimidyl 3-(2- pyridyldithio)propionate (SPDP), long chain-SPDP, maleimidobenzoic acid-N- hydroxysuccinimide ester (MBS), succinimidyl trans-4-(maleimidylmethyl)cyclohexane-1- carboxylate (SMCC), succinimidyl iodoacetate (SIA) and 6-aminohexanoic acid (Ahx).

[0124] In certain embodiments a linker, if present, may be a derivative of a diamine. A diamine moiety or derivative can provide a linker arm of varying lengths and chemistries for chemically linking molecules by derivatizing, optionally, with carboxylic acids. Non-limitingexamples of diamines include ethylenediamine (EDA), propylenediamine, spermidine, spermine, hexanediamine, and diamine-amino acids, such as homolysine, lysine, ornithine, diaminobutyric acid and diaminopropionic acid. In other embodiments, moieties of an imaging agent can be chemically linked to a dicarboxylic acid, for example, succinic acid, glutaric acid, suberic acid, or adipic acid. In one embodiment, the linker is aminoethylmaleimide.

[0125] In certain embodiments, a linker can be formed from an azide moiety that can react with substituted alkynes in an azide-acetylene Huisgen [3+2] cycloaddition. In certain embodiments the azide or alkyne linker can link a polyethyleneglycol (PEG) moiety to, for example, an enzymatically cleavable oligopeptide. Other contemplated linkers include propargylglycine, pentanoyl, pentynoic acid, propargylic acid, and / or propargylamine moieties.

[0126] In some embodiments, polyethylene glycol (PEG) modification can be used as linkers. PEGylation can enhance peptide solubility, improving handling and formulation. Moreover, PEGylation can confer increased stability by protecting against enzymatic degradation, thus extending peptide half-life. Cellular uptake can be improved due to increased endocytosis and evasion of efflux transporters, while overall toxicity can be lowered. PEGylation can also enhance water solubility of peptides while simultaneously reducing their renal clearance. The pharmacokinetics of the agent can be modulated based on PEG chain length, allowing for tailored release rates, biodistribution, and elimination kinetics. In some embodiments, a direct conjugation linkers can be used, where the imaging molecule is affixed directly to the peptide. Another embodiment includes the use of peptidic linkers, composed of short sequences of amino acids, providing biodegradability and the flexibility to be designed with specific properties. In other embodiment, carbohydrate- based linkers can be utilized, demonstrating biocompatibility and ease of synthesis with the possibility of modification to adjust their properties. Another embodiment can use ester and amide linkers, with amide linkers providing notable stability and resistance to breakdown in the body, and ester linkers engineered to cleave under specific conditions. Additional embodiments involve disulfide linkers, designed to be cleaved by glutathione, a reducing agent found in cells, enabling selective release of an imaging molecule in certain bodily areas. Heterobifunctional linkers, possessing two different functional groups to allow connection of two dissimilar types of molecules, are another embodiment. Enzyme-cleavable linkers can also be used, designed to cleave in response to specific enzymes, providing a mechanism for the controlled release of the imaging molecule. These linkers allow for the optimization of desired biodistribution and pharmacokinetics, and the intended use of the imaging agent. These embodiments offerinnovative strategies in the design and application of peptide imaging agents, enhancing efficacy and adaptability for a myriad of imaging, clinical and diagnostic applications.

[0127] In certain embodiments, fluorophores, quenchers, or other reporters are directly linked to the imaging agent using reactive NHS ester groups on the fluorophores, quenchers, or reporters which react with an amine group on the enzymatically cleavable oligopeptide. In certain other embodiments, carboxylic acid groups on the fluorophores, quenchers, or other reporters can be activated in situ by activating agents known in the art, such as 2-(1 H- benzotriazole-1-yl)-1 ,1 ,3,3,-tetramethyluronium hexafluorophosphate (HBTU), 1-ethyl-3-(3'- dimethylaminopropyl)-carbodiimide hydrochloride (EDC), N,N'-dicyclohexylcarbodiimide (DCC), N,N’-disuccinimidyl carbonate (DSC). In other embodiments, reporters including a sulfhydryl or thiol group, can be chemically linked to the agent via a bifunctional cross-linker that has a second moiety that can react with a sulfhydryl (thiol) group. Such crosslinking agents include, for example and as described above, SPDP, long chain-SPDP, SIA, MBS, SMCC, and others that are well known in the art.

[0128] Useful linker moieties include both natural and non-natural amino acids, oligopeptides, for example, linear or cyclic oligopeptides, and nucleic acids.

[0129] The linker can be a peptide or peptide moiety which optionally includes a proteolytic or non-proteolytic cleavage site, such as anester linkage, that can be cleaved due to pH changes at the site of interest.Imaging Guided Surgery

[0130] Disclosed herein are methods of use of the disclosed imaging agents for image guided surgery. In some embodiments, various technologies are utilized for image-guided surgery, enabling the enhancement of diagnosis, precision, and accuracy during surgical procedures. In some embodiments, the surgeon can use the disclosed imaging agents to diagnose staging of endometriosis and cancer. In some embodiments, the surgeon can use the disclosed imaging agents to detect the location of the endometriosis lesion or cancer tumor on which organ system. In some embodiments, the surgeon can use the disclosed imaging agents to identify and / or stage the disease. In some embodiments, the surgeon can use the disclosed imaging agents to help with excision of lesion or tumor.

[0131] In some embodiments, the surgeon can use the disclosed imaging agents to help with excision of lesion or tumor. In some embodiments, this includes fluorescence-guided surgery (FGS). FGS uses fluorescent probes or dyes that are either ingested or injected into the patient, which then accumulate in the tumor. When exposed to a specific wavelength of light, these probes or dyes emit fluorescence, which can be detected by a specialized camera systemand viewed by the surgeon, aiding in the visualization of the tumor. The imaging system used to visualize the fluorescence in FGS typically includes a light source to excite the fluorescent compound, a camera to capture the emitted fluorescence, and a display monitor for the surgeon to view the images. For instance, the “da Vinci Surgical System” has been upgraded to include a near-infrared fluorescence imaging system for real-time, image-guided surgery. SPY Elite System, Artemis and Artemis Pro (Quest Medical Imaging), and handheld devices like Fluobeam (Fluoptics), Hamamatsu’s PDE-Neo, Visee PDT Imaging System (Visee Medical): used for visualizing fluorescence during photodynamic therapy (PDT) In some embodiments, a handheld probe as part of a surgical microscope system (Leica FL560 (Leica Microsystems)) can be used to visualize fluorescence in areas that might be difficult to reach with the microscope.

[0132] In some embodiments, the disclosed imaging agents can also be visualized by instruments such as LightPath Imaging System (Lightpoint Medical), a handheld device is designed to detect Cerenkov Luminescence, which is emitted by certain types of radiotracers including Carbon-11 (11C), Nitrogen-13 (13N), Oxygen-15 (15O), Copper-64 (64Cu), Gallium-68 (68Ga), Fluorine-18 (18F).

[0133] Herein describes a method for identifying, visualizing, diagnosing and surgically removing diseased tissue. The method involves the application of an imaging agent to a patient, which facilitates the visualization of diseased tissue either by the naked eye or with the assistance of a machine. In some embodiments, an image capturing the areas of glowing diseased tissue is acquired prior to the surgical procedure. In some embodiments, this process is carried out in real time. In some embodiments, the captured image is displayed to the surgeon. In some embodiments the image is a still image or series of still images. In some embodiments the image is a video. This image is then presented to the surgeon during the surgery using a reader device, allowing the surgeon to visually identify the glowing diseased tissue in the patient. The surgeon can directly observe the identified glowing tissue or use a specialized visualization machine for assistance. Based on the identified glowing diseased tissue, the surgeon can make a surgical diagnosis, staging, and / or proceed with the excision of the diseased tissue accordingly.

[0134] In some embodiments, the agents are visualized by Magnetic Resonance Imaging (MRI). In some embodiments, Computed Tomography (CT) can be used. In additional embodiments, imaging methods such as X-ray and Single Photon Emission Computed Tomography (SPECT) may be employed. In other embodiments, ultrasound imaging can be used. In some embodiments, the peptides can be utilized in optical imaging, which uses light toprobe the molecular makeup of tissues, aiding surgeries where the tumor or target is near the surface. In some embodiments make use of Positron Emission Tomography (PET), to guide specific surgical procedures. In some embodiments, PET is used in combination with CT.

[0135] In some embodiments, the agents are visualized with MRI machines with lower magnetic field strengths, which are less expensive and ineffective with gadolinium contrast.

[0136] In some embodiments, the agents are visualized with a 0.064T Portable MRI Machine. These machines are designed for point-of-care use, making MRI technology more accessible and affordable. They can be used beside the patient's bed and are particularly useful in settings where traditional MRI access is limited. While offering less detail due to their low field strength, they provide significant value in specific clinical situations, such as stroke diagnosis in emergency settings.

[0137] In some embodiments, the agents are visualized with a low-field MRI Machine (0.2T - 0.4T). These machines are suitable for larger patients or those with claustrophobia due to their open design. These machines have lower costs but produce images with less detail. They are typically used for extremity imaging, with some capable of full-body scans.

[0138] In some embodiments, the agents are visualized with a 1.5T MRI Machine. These machines are the standard in clinical imaging, offering a balance between image quality, speed, and cost. They are versatile, suitable for a broad range of diagnostic purposes including brain, spine, and musculoskeletal imaging.

[0139] In some embodiments, the agents are visualized with a 3T MRI Machine. These machines provide higher image quality and detail, useful for advanced imaging needs such as detailed brain, musculoskeletal studies, and high-resolution imaging. Though more expensive, they can improve patient throughput with faster imaging times.

[0140] In some embodiments, the agents are visualized with a Ultra High-field MRI Machines (7T and above). These machines are mainly used in research, these scanners offer exceptional detail for medical research. Beneficial for imaging tiny structures and functional MRI, but not commonly used in standard clinical practice due to their high cost and technical requirements.

[0141] In some embodiments, the agents are visualized with an Open MRI Machine. These machines are designed to reduce claustrophobia and accommodate larger patients, these machines have a lower field strength (around 0.5T), resulting in less detailed images than closed-bore machines but providing a more comfortable experience for certain patients.

[0142] In some embodiments, the agents are visualized with a Front-Open MRI Machine. These machines are similar to open MRIs, but designed for increased comfort andaccess, particularly for exams requiring specific positioning. They offer an alternative for patients with claustrophobia or mobility issues.

[0143] MRI machines can generally use contrast agents to enhance image quality, with the choice of machine depending on imaging needs, patient considerations, and costeffectiveness. The 1.5T and 3T machines are most commonly accessible for clinical use outside of dedicated imaging centers due to their versatility and balance of image quality, speed, and cost.

[0144] In some embodiments, the agents are visualized with a full-body scan, i.e. not just a pelvic scan, to diagnose all subtypes and stages of endometriosis lesions on any organ system. This can provide both the patient and the clinical provider with greater insights into disease progression and prognosis for data-driven care. For example, endometriosis on the bladder can present itself as painful UTIs while endometriosis on the appendix can present itself as excruciating appendicitis. Critically, this MRI approach can also enable the detection and characterization of malignancy, providing critical information for patients so that they can receive appropriate care. For example and without limitation, a full body scan can be used to detect endometriosis lesions in the appendix, brain, spine, thyroid, lung, liver, gallbladder, pancreas, spleen, kidney, adrenal glands, bladder, ovaries, uterus, bowels, or any combination thereof.

[0145] In one embodiment, the present disclosure discloses the utilization of an imaging agent during endometriosis surgery. In certain embodiments, the pharmaceutical agent is introduced into the peritoneal cavity contemporaneously with the surgical procedure. In alternative embodiments, the administration of the pharmaceutical agent takes place prior to the surgical intervention, no less than 15 minutes, 30 minutes, 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 prior to surgery.

[0146] Furthermore, in additional embodiments, the pharmaceutical agent may be administered through alternative routes, encompassing, but not limited to, intravenous (IV), subcutaneous (SC), vaginal, rectal, oral, and intramuscular (IM) routes. By employing this method, the surgeon gains the ability to visualize endometriosis lesions with remarkable precision, facilitating their identification in terms of size, location, and disease staging. This visualization can be accomplished either by the naked eye or through the utilization of a handheld tool or imaging equipment. Armed with this enhanced visual information, the surgeon can then proceed to perform a meticulous excision of the endometriosis lesion while minimizingthe risk of inadvertently excising healthy tissue. This technique represents a significant advancement in endometriosis surgery, enabling improved surgical outcomes and patient care.

[0147] This can be applied to surgeries involving diseases with [3-catenin misregulation, including but not limited to colorectal cancer, hepatocellular carcinoma, endometrial cancer, ovarian cancer, medulloblastoma, glioblastoma, melanoma, and Alzheimer’s disease. In some embodiments, these agents can be utilized for diagnostic, imaging, and treatment of gynecological conditions include endometriosis, uterine fibroids, adenomyosis, ovarian cysts, polycystic ovary syndrome (PCOS), endosalpingiosis and cervical, uterine, and vaginal polyps, and / or its symptoms.Pharmaceutical Compositions

[0148] Disclosed herein are peptides and peptidomimetic agents comprising an amino acid sequence of any one of SEQ ID NO: 1-SEQ ID NO: 500, or having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to any one of SEQ ID NO: 1-SEQ ID NO: 499, in a pharmaceutically acceptable carrier.

[0149] In some embodiments, the agents and / or peptides described herein are formulated into pharmaceutical compositions. Pharmaceutical compositions are formulated in a conventional manner using one or more pharmaceutically acceptable inactive ingredients that facilitate processing of the active agents and / or peptides into preparations that are used pharmaceutically. Proper formulation is dependent upon the route of administration chosen. A summary of pharmaceutical compositions described herein is found, for example, in Remington: 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, Pa. 1975; Liberman, H. A. and Lachman, L, Eds., Pharmaceutical Dosage Forms, Marcel Decker, New York, N.Y., 1980; and Pharmaceutical Dosage Forms and Drug Delivery Systems, Seventh Ed. (Lippincott Williams & Wilkinsl 999), herein incorporated by reference for such disclosure.

[0150] In some embodiments, the agents and / or peptides described herein are administered either alone or in combination with pharmaceutically acceptable carriers, excipients or diluents, in a pharmaceutical composition. Administration of the agents and / or peptides and compositions described herein can be affected by any method that enables delivery of the agents and / or peptides to the site of action. These methods include, though are not limited to delivery via enteral routes (including oral, gastric or duodenal feeding tube, rectal suppository and rectal enema), parenteral routes (injection or infusion, including intraarterial, intracardiac, intradermal, intraduodenal, intramedullary, intramuscular, intraosseous,intraperitoneal, intrathecal, intravascular, intravenous, intravitreal, epidural and subcutaneous), inhalational, transdermal, transmucosal, sublingual, buccal and topical (including epicutaneous, dermal, enema, eye drops, ear drops, intranasal, vaginal, and intrauterine) administration, although the most suitable route may depend upon for example the condition and disorder of the recipient. By way of example only, agents and / or peptides described herein can be administered locally to the area in need of treatment, by for example, local infusion during surgery, topical application such as creams or ointments, injection, catheter, implant, or inserted device. The administration can also be by direct injection at the site of a diseased tissue or organ.

[0151] In some embodiments, the pharmaceutical composition is in solution form. In some embodiments the formulation utilizes buffers such as sodium acetate, histidine buffer. In some embodiments cosolvents of NMP, DMSO, Polypropylene glycol are used. In some embodiments, aggregation suppressors such as PS80 are used. In some embodiments, NaOleate, 2-phenoxyethanol, niacinamide, HPBCD, and trehalose are used as excipients. In some embodiments, the formulation is an aqueous suspension. In some embodiments, Na- CMC, PS80, PBS, and trehalose are utilized as excipients.

[0152] In some embodiments, the pharmaceutical composition is an oil suspension. In some embodiments, the oil is cottonseed, corn, olive, sesame and / or castor oil. In some embodiments, the formulation is a water-in-oil or oil-in-water emulsion. In some embodiments, it consists of a mixture of cotton:soy:olive, corn and cotton, sesame and cotton oil. In some embodiments, emulsifiers are used (including but not limited to PS80, PS20, Kolliphor EL, Lecethin, or some combination). In some embodiments, NMP, Ethanol, propylene glycol, and DMSO can be used as solvents.

[0153] 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 a suspension in an aqueous liquid or a non-aqueous liquid; or as an oil-in-water liquid emulsion or a water-in-oil liquid emulsion. In some embodiments, the active ingredient is presented as a bolus, electuary or paste.

[0154] 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 a plasticizer, 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 a powder or granules,optionally mixed with binders, inert diluents, or lubricating, surface active or dispersing agents. Molded tablets may be made by molding in a suitable machine a mixture of the powdered agent and / or peptide moistened with an inert liquid diluent. In some embodiments, the tablets are coated or scored and are formulated so as to provide slow or controlled release of the active ingredient therein. All formulations for oral administration can be in dosages suitable for such administration. The push-fit capsules can contain the active ingredients in admixture with filler such as lactose, binders such as starches, and / or lubricants such as talc or magnesium stearate and, optionally, stabilizers. In soft capsules, the active agents and / or peptides may be dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin, or liquid polyethylene glycols. In some embodiments, stabilizers are added. Dragee cores are provided with suitable coatings. For this purpose, concentrated sugar solutions may be used, which may optionally contain gum arabic, talc, polyvinyl pyrrolidone, carbopol gel, polyethylene glycol, and / or titanium dioxide, lacquer solutions, and suitable organic solvents or solvent mixtures. Dyestuffs or pigments may be added to the tablets or dragee coatings for identification or to characterize different combinations of active agent and / or peptide doses.

[0155] In some embodiments, pharmaceutical compositions are formulated for parenteral administration by injection, e.g., by bolus injection or continuous infusion. Formulations for injection may be presented in unit dosage form, e.g., in ampoules or in multidose containers, with an added preservative. The compositions may take such forms as suspensions, solutions or emulsions in oily or aqueous vehicles, and may contain formulator agents such as suspending, stabilizing and / or dispersing agents. The compositions may be presented in unit-dose 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 the sterile liquid carrier, for example, saline or sterile pyrogen-free water, immediately prior to use. Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules and tablets of the kind previously described.

[0156] Pharmaceutical compositions for parenteral administration include aqueous and non-aqueous (oily) sterile injection solutions of the active agents and / or peptides which may contain antioxidants, buffers, bacteriostats and solutes which render the formulation isotonic with the blood of the intended recipient; and aqueous and non-aqueous sterile suspensions which may include suspending agents and thickening agents. Suitable lipophilic solvents or vehicles include fatty oils such as sesame oil, or synthetic fatty acid esters, such as ethyl oleate or triglycerides, or liposomes. Aqueous injection suspensions may contain substances which increase the viscosity of the suspension, such as sodium carboxymethyl cellulose, sorbitol, ordextran. Optionally, the suspension may also contain suitable stabilizers or agents which increase the solubility of the agents and / or peptides to allow for the preparation of highly concentrated solutions.

[0157] Pharmaceutical compositions may also be formulated as a depot preparation. Such long acting formulations may be administered by implantation (for example subcutaneously or intramuscularly) or by intramuscular injection. Thus, for example, the agents and / or peptides may be formulated with suitable polymeric or hydrophobic materials (for example, as an emulsion in an acceptable oil) or ion exchange resins, or as sparingly soluble derivatives, for example, as a sparingly soluble salt.

[0158] For buccal or sublingual administration, the compositions may take the form of tablets, lozenges, pastilles, or gels formulated in conventional manner. Such compositions may comprise the active ingredient in a flavored basis such as sucrose and acacia or tragacanth.

[0159] Pharmaceutical compositions may also be formulated in rectal or vaginal compositions such as suppositories or retention enemas, e.g., containing conventional suppository bases such as cocoa butter, polyethylene glycol, or other glycerides. When inserted, the suppository base liquefies or becomes water-miscible at body temperature so as to allow the components to become in contact with the mucous membrane for a sufficient period of time to have an imaging, diagnostic, and / or therapeutic effect. The weight percent of the suppository base is dependent upon the size of the bodily orifice of the human and / or the animal, the dosage composition necessary to have an imaging, diagnostic, and / or therapeutic effect, and its physiochemical characteristics that allow it to remain solid at or below room temperature. In some embodiments, the suppository comprises from about 50% to greater than 99%, or about 75% to greater than 99% by weight of the suppository base. In some embodiments, the suppository comprises about 75% to about 98% by weight polyethylene glycol. In some embodiments, the suppository comprises about 2% to about 25% by weight polysorbate. The suppository base has a molecular weight in the range of about 400 to about 5000, or about 950 to about 3700 (US 2009 / 0311290, incorporated by reference herein for such disclosure).

[0160] In some cases, the pharmaceutical composition comprises an absorption enhances, such as sodium caprate. Some of such cases include compositions and dosage forms for use in the rectum or vagina as described herein.

[0161] Pharmaceutical compositions may be administered topically or rectally, that is, by non-systemic administration. This includes the application of an agent and / or peptide of the present disclosure externally to the epidermis or the buccal cavity and the instillation of such anagent and / or peptide into the rectum or vagina such that the agent and / or peptide does not significantly enter the blood stream. In contrast, systemic administration refers to oral, intravenous, intraperitoneal and intramuscular administration.

[0162] Pharmaceutical compositions suitable for topical administration include liquid or semi-liquid preparations suitable for penetration through the skin such as solutions, lotions, shake lotions, creams, ointments, gels, foams, transdermal patches, powders, solids, sponges, tapes, vapors, pastes, tinctures, microparticles, microcapsules, nanoparticles, liposomes, or emulsions, including those suitable for delivery to the vagina or rectum. The active ingredient may comprise, for topical administration, from 0.001 % to 10% w / w, for instance from 1% to 2% by weight of the formulation.

[0163] Pharmaceutical compositions for administration by inhalation are conveniently delivered from an insufflator, nebulizer pressurized packs or other convenient means of delivering an aerosol spray. Pressurized packs may comprise a suitable propellant such as dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas. In the case of a pressurized aerosol, the dosage unit may be determined by providing a valve to deliver a metered amount. Alternatively, for administration by inhalation or insufflation, pharmaceutical preparations may take the form of a dry powder composition, for example a powder mix of the agent and / or peptide and a suitable powder base such as lactose or starch. The powder composition may be presented in unit dosage form, in for example, capsules, cartridges, gelatin or blister packs from which the powder may be administered with the aid of an inhalator or insufflator.

[0164] In some embodiments, an agent disclosed herein is formulated in such a manner that delivery of the agent and / or peptide to a particular region of the gastrointestinal tract is achieved. For example, an agent and / or peptide disclosed herein is formulated for oral delivery with bioadhesive polymers, pH-sensitive coatings, time dependent, biodegradable polymers, microflora activated systems, and the like, in order to effect delivering of the agent and / or peptide to a particular region of the gastrointestinal tract.

[0165] In some embodiments, an agent and / or peptide disclosed herein is formulated in such a manner that delivery of the agent and / or peptide to a particular region of the urogenital or anorectal mucosa is achieved. For example, an agent and / or peptide disclosed herein is formulated for intravaginal delivery with bioadhesive polymers, pH-sensitive coatings, time dependent, biodegradable polymers, microflora activated systems, and the like, in order to effect delivering of the agent and / or peptide to a particular region of the urogenital system. In some embodiments, an agent and / or peptide disclosed herein is formulated to provide acontrolled release of the agent and / or peptide. Controlled release refers to the release of the agent and / or peptide described herein from a dosage form in which it is incorporated according to a desired profile over an extended period of time. Controlled release profiles include, for example, sustained release, prolonged release, pulsatile release, and delayed release profiles. In contrast to immediate release compositions, controlled release compositions allow delivery of an agent to a subject over an extended period of time according to a predetermined profile. Such release rates can provide imaging and / or therapeutically effective levels of agent for an extended period of time and thereby provide a longer period of pharmacologic response while minimizing side effects as compared to conventional rapid release dosage forms. Such longer periods of response provide for many inherent benefits that are not achieved with the corresponding short acting, immediate release preparations.

[0166] Approaches to deliver the intact therapeutic, imaging and / or diagnostic agent to the particular regions of the urogenital system (such as the vagina) or gastrointestinal tract (e.g., such as the colon), include: (i) Coating with polymers: The intact molecule can be delivered to the colon without absorbing at the upper part of the intestine by coating of the drug molecule with the suitable polymers, which degrade only in the colon. In addition, coating with polymers can provide protection or controlled-release profile for vaginal formulations; (ii) Coating with pH- sensitive polymers: Enteric, colon, and vaginal targeted delivery systems can be based on the coating of tablets or pellets, which are filled into conventional hard gelatin capsules. 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 S 100 are copolymers of methacrylic acid and methyl methacrylate; (iii) Coating with biodegradable polymers; (iv) Embedding in matrices; (v) Embedding in biodegradable matrices and hydrogels; (vi) Embedding in pH-sensitive matrices; (vii) Timed release systems; (viii) Redox-sensitive polymers; (ix) Bioadhesive systems; (x) Coating with microparticles; and (xi) Osmotic controlled drug delivery.

[0167] Another approach towards vaginal- and colon-targeted drug delivery or controlled-release systems includes embedding the drug in polymer matrices to trap it and release it in the vagina or colon. These matrices can be pH-sensitive or biodegradable. Matrix- Based Systems, such as multi-matrix (MMX)-based delayed-release tablets, ensure the drug release in the vagina or colon.

[0168] The compositions and methods described herein can include delivering the therapeutic, imaging, and / or diagnostic agents via a variety of dosage forms and devices, including those described above and below. Many of the dosage forms described below providefor several advantages. These can include local delivery of the therapeutic, imaging, and / or diagnostic agents and / or peptides. Local delivery can reduce the side effects that can sometimes be associated with systemic delivery and off-target effects. In some cases, the dosage forms and devices described herein can increase patient compliance including, for example, by providing long-term or continuous delivery of the agents and / or peptides described herein. In some cases, these dosage forms and devices can also increase tolerability and safety.

[0169] Peptides and compositions described herein can be incorporated in an intravaginal ring for delivery. Delivery of therapeutic, imaging, and / or diagnostic via intravaginal ring (IVR) allows for local delivery, increasing safety and tolerability of the therapeutic, imaging, and / or diagnostic substance. IVRs additionally can offer multiple advantages: bypass gastrointestinal absorption and hepatic / renal first-pass metabolism, lower effective dosage, continuous delivery and / or controlled release profiles, extended time between doses, reduced side effects, low serum drug concentrations, patient self-administration, and improved patient satisfaction. IVRs can successfully deliver drugs, including hydrophilic and macromolecular agents, to organs affected by EMS, including deep infiltrating EMS (DIE). The biodistribution of the vaginal ring delivery system is via tissue absorption rather than serum and can reach all organs that are affected by EMS in the peritoneal cavity.

[0170] Vaginal rings usually consist of an inert elastomer ring coated by another layer of elastomer containing the drug to be delivered. The rings can be easily inserted, left in place for the desired period of time, then removed by the user. The ring may be solid or hollow containing the therapeutic, imaging, and / or diagnostic component, or it may be a porous material releasing the drug therefrom. The ring can optionally include a third, outer, rate-controlling elastomer layer which contains no drug. Optionally, the third ring can contain a second drug for a dual release ring. The drug can be incorporated into polyethylene glycol throughout the silicone elastomer ring to act as a reservoir for drug to be delivered. In some cases, the IVR can comprise silicone, compressed tablets, or lyophilized gel.

[0171] Pessaries, cups, strips, tablets, and suppositories are other examples of drug delivery systems which can be used in the present disclosure. These systems have been used for delivery of vaginal contraceptives and have been described extensively in the literature.

[0172] Another example of a delivery system is the vaginal sponge and foams. The desired pharmaceutical agent can be incorporated into a silicone matrix which is coated onto a cylindrical drug-free polyurethane vaginal sponge, as described in the literature.

[0173] In some embodiments, the IVR comprises peptides or compositions as described herein, formulated as a suppository, solution, lotion, shake lotion, cream, ointment, gel, foam, transdermal patch, powder, solid, sponge, tape, paste, tincture, emulsion, microparticle, microcapsule, nanoparticle, liposome or a capsule containing microparticles, microcapsules, nanoparticles, or liposomes. IVRs have further been modified to comprise a variety of delivery vehicles, such as silicone inserts, compressed tablets, or lyophilized gel, to optimize the release profile of hydrophilic or high molecular weight agents such as peptides, proteins, Or antibodies (Morrow, et. al., Eur J Pharm Biopharm, 2011 January; 77(1): 3-10, incorporated by reference herein for such disclosure). IVRs can also comprise one or more absorption enhancers, such as sodium caprate.

[0174] In some embodiments, IVRs described herein are formulated to comprise from about 0.01 mg to about 5000 mg agent and / or peptide. In some embodiments, the IVR can 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 agent and / or peptide.

[0175] In some embodiments, IVRs described herein are formulated to deliver from about 0.01 mg to about 1000 mg agent and / or peptide / 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 agent and / or peptide / day.

[0176] Peptides and compositions described herein can be incorporated in a tampon device for delivery. A tampon device typically comprises a vaginal tampon having a proximal end and a distal end. A cup-shaped porous foam portion at the distal end fits around the cervix and contains a peptide or composition as described herein for delivery. The device may also include a nonabsorbing axial tube having a distal opening and extending through the porous foam cup into the tampon for conducting blood flow to the absorbent material. Optionally, a retrieval string or tape connected to the tampon device is also included. The absorbent vaginal tampon contains peptides or compositions as described herein or may be coated with the peptides or compositions as described herein and be used as a medicated tampon for delivery.

[0177] Peptides and compositions described herein may be incorporated in to a solid for local delivery. A solid may be in the form of a pessary or vaginal or rectal suppository. The solid dosage form may melt when it reaches body temperature. Alternatively, the solid may retain its structure and release incorporated compositions as described herein. The solid may be a pessary designed to provide support within the vagina. A vaginal sponge may be embedded with a composition described herein for intravaginal delivery.

[0178] Peptides and compositions described herein can be incorporated in a topical formulation for delivery. A topical medication is applied to a body surface, such as the skin or a mucous membrane. In some instances, the body surface includes, without limitation, epithelial tissue, mucosal tissue, peritoneum, perimetrium, and endometrium. Agent and / or peptide is absorbed through the body surface to achieve a local or systemic effect. Topical medications are optionally formulated in the following classes: a topical solution; a lotion; a shake lotion; a cream; an ointment; a gel; a foam; a transdermal patch; a powder; a solid; sponge; tape; vapor; paste; or tincture. A topical solution may be administered as a rinse, spray, or drop, typically with low viscosity with water or alcohol in the base. A lotion can be thicker and more emollient than a solution. It is usually an oil mixed in water and may have less alcohol than a solution. A shake lotion is a mixture that separates into two or three parts with time. It may be an oil mixed with a water-based solution that needs to be shaken into suspension prior to use. A cream is an emulsion of oil and water in approximately equal proportions. An ointment is a homogeneous, viscous, semi-solid preparation, most commonly a greasy, thick oil (oil 80%-water 20%) with a high viscosity. An ointment may comprise a hydrocarbon base, an absorption base, a water- soluble base, an emulsifying base, or a vegetable oil, or any combination thereof. An ointment is formulated in a base which may include, but is not limited to, a hydrocarbon base, such as hard paraffin, soft paraffin, microcrystalline wax, or ceresine; absorption bases, such as wool fat or beeswax; water-soluble bases, such as macrogols 200, 300, or 400; emulsifying bases, such as emulsifying wax or cetrimide; or vegetable oils, such as olive oil, coconut oil, sesame oil, almond oil, or peanut oil.

[0179] A gel is a semisolid emulsion. Non-limiting examples of useful emulsifiers include acrylic acid polymers (such as carbomer brand thickeners e.g. Carbomer 934P, manufactured by Voveon, Inc.), polyoxyethylene- 10-stearyl ether, polyoxyethylene-20-stearyl ether, cetostearyl alcohol, cetyl alcohol, cholesterol, diglycol stearate, glyceryl monostearate, glyceryl stearate, polygeyceryl-3-oleate, hydroxypropyl cellulose, hydroxypropylmethyl cellulose, lanolin, polyoxyethylene lauryl ether, methyl cellulose, polyoxyethylene stearate, polysorbate, propylene glycol monostearate, sorbitan esters, stearic acid or mixtures of two or more thereof.

[0180] The amount of emulsifier in the topical formulation can range from about 1 to about 40 weight percent, and in some embodiments from about 5 to about 30 weight percent, on a basis of total weight of the topical formulation.

[0181] The gel formulations as described herein may include one or more gelling agents. Non-limiting examples of useful gelling agents include carboxylic acid polymers including acrylic acid polymers crosslinked with cross links such as allyl ethers of sucrose (e.g., carbomer brand thickeners), cetostearyl alcohol, hydroxymethyl cellulose, polyoxyethylenepolyoxypropylene copolymer, sodium carboxymethylcellulose, polyvinyl pyrrolidone, or mixtures of two or more thereof

[0182] The amount of gelling agent in the topical gel formulation can range from about 0.1 to about 10 weight percent, and in some embodiments from about 0.1 to about 1 weight percent, on a basis of total weight of the topical formulation.

[0183] The gel formulations described herein can further comprise one or more alkalinizers, for example sodium hydroxide, in amount of less than about 2 weight percent as activators of gelling.

[0184] The formulations can contain one or more additional excipients well known in the art, for example water and a thickening agent such as colloidal silicon dioxide.

[0185] A thermoreversible gel is a liquid formulation that turns to gel once inserted into the rectum or vagina. A thermoreversible gel allows for easier administration and positioning than conventional suppositories or pessaries and can prevent dosage form leakage. It is formulated as a polymer solution consisting of thermoreversible polymers (e.g., poloxamers, in combination with mucoadhesive polymers that enable gel attachment to the mucosa). In situ thermoreversible liquid-gel formulations, also called thermoreversible “liquid suppositories”, are liquid at low temperatures (<10C) and turn to gel at body temperature.

[0186] Active agent can be incorporated into a surgical tape for an occlusive dressing. Medication can be applied as an ointment or gel, to reach a mucous membrane through vaporization. A paste combines oil, water, and a powder. A tincture typically contains a high percentage of alcohol for application to the skin.

[0187] Peptides and compositions described herein can be incorporated in a transdermal patch delivery. A transdermal patch provides a controlled release of medication either through a porous membrane covering a reservoir of medication or through body heat melting thin layers of medication embedded in the patch adhesive.

[0188] Compositions described herein may be incorporated in a film for delivery. Films are thin, small polymeric formulations that can be easily inserted into the vaginal cavity withoutan applicator and without causing discomfort Vaginal films are easier to apply than other types of vaginal formulations such as pessaries, foams and gels (Rohan L C et al., AAPS Journal 2009; 11 :78-87, incorporated by reference herein for such disclosure).

[0189] Peptides and compositions described herein can be incorporated in an intrauterine device (mall often T-shaped device that is inserted into the uterus for delivery. IUDs typically contain copper, progestogen, or levonorgestrel. Compositions as described herein are incorporated into an IUD device to be released slowly over time.

[0190] Peptides and compositions as described herein can be formulated for systemic delivery. Parenteral injections can be formulated for bolus injection or continuous infusion. The pharmaceutical compositions can be in a form suitable for parenteral injection as a sterile suspension, solution or emulsion in oily or aqueous vehicles, and can contain formulation agents such as suspending, stabilizing and / or dispersing agents. Pharmaceutical formulations for parenteral administration include aqueous solutions of a peptide described herein in water soluble form. Suspensions of peptides described herein can be prepared as oily injection suspensions. Suitable lipophilic solvents or vehicles include fatty oils such as sesame oil, or synthetic fatty acid esters, such as ethyl oleate or triglycerides, or liposomes. Aqueous injection suspensions can contain substances which increase the viscosity of the suspension, such as sodium carboxymethyl cellulose, sorbitol, or dextran. The suspension can also contain suitable stabilizers or agents which increase the solubility and / or reduce the aggregation of such peptides described herein to allow for the preparation of highly concentrated solutions. Alternatively, the peptides described herein can be lyophilized or in powder form for reconstitution with a suitable vehicle, e.g., sterile pyrogen-free water, before use. In some embodiments, a purified peptide is administered intravenously.

[0191] Peptides and compositions described herein can be formulated for oral delivery. Pharmaceutical compositions which can be used orally include tablets, push-fit capsules made of gelatin, as well as soft, sealed 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 may be prepared by compressing in a suitable machine the active ingredient in a free-flowing form such as a powder or granules, optionally mixed with binders, inert diluents, or lubricating, surface active or dispersing agents. Molded tablets may be made by molding in a suitable machine a mixture of the powdered agent and / or peptide moistened with an inert liquid diluent. In some embodiments, the tablets are coated or scored and are formulated so as to provide slow or controlled release of the active ingredient therein. Formulations for oral administration may be in dosages suitable for such administration.The push-fit capsules can contain the active ingredients in admixture with filler such as lactose, binders such as starches, and / or lubricants such as talc or magnesium stearate and, optionally, stabilizers. In soft capsules, the active agents and / or peptides may be dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin, or liquid polyethylene glycols. In some embodiments, stabilizers are added. Dragee cores are provided with suitable coatings. For this purpose, concentrated sugar solutions may be used, which may optionally contain gum arabic, talc, polyvinyl pyrrolidone, carbopol gel, polyethylene glycol, and / or titanium dioxide, lacquer solutions, and suitable organic solvents or solvent mixtures. Dyestuffs or pigments may be added to the tablets or Dragee coatings for identification or to characterize different combinations of active agent and / or peptide doses.

[0192] Additional pharmaceutical approaches to targeted delivery of therapeutics to particular regions of the gastrointestinal tract are known. Chourasia M K, Jain S K, Pharmaceutical approaches to colon targeted drug delivery systems., J Pharm Sci. 2003 January-April; 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 July; 5(3):186-98. Van den Mooter G. Colon drug delivery. Expert Opin Drug Deliv. 2006 January; 3(1): 111-25. Seth Amidon, Jack E. Brown, and Vivek S. Dave, Colon-Targeted Oral Drug Delivery Systems: Design Trends and Approaches, AAPS Pharm Sci Tech. 2015 August; 16(4): 731-741. Each of these references is incorporated by reference herein for such disclosure.

[0193] It should be understood that in addition to the ingredients particularly mentioned above, the agents and / or peptides and compositions described herein may include other agents conventional in the art having regard to the type of formulation in question. For example, agents suitable for oral administration may include flavoring agents.

[0194] Peptides and compositions described herein may contain additional excipients to improve performance of the imaging, diagnostic and / or therapeutic. Compositions may contain absorption enhancers, or permeation enhancers, to improve absorption across the epidermal or mucosal surface and membrane permeation. Enhancers formulated in compositions described herein may include, but are not limited to, sulphoxides, such as dimethyl sulphoxides (DMSO); laurocapran (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-hydroxylbenzoyl)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 caprylate; protease inhibitor and omega-3 fatty acid; liquidmixed-micelle spray; lipid polymer micelle; alkylglycosides; chitosan; dodecyl-2-N,N- dimethylamino propionate (DDAIP); cell-membrane-lipid components; nanoparticles; liposomes, ligands; and lipophilic modifications.

[0195] Peptides and compositions described herein may be formulated as a solution, a lotion, a shake lotion, a cream, an ointment, a gel, a foam, a mucoadhesive composition, an emulsion, liposomes, a coating, a core, a matrix, a lyophilisate.Methods of Administration

[0196] In some embodiments, the agent and / or peptide compositions described herein are used in the preparation of medicaments for the imaging, diagnosis, and / or treatment of diseases, conditions, or symptoms in a mammal that would benefit from administration of a p- catenin-agent and / or peptide. Methods for treating any of the diseases or conditions described herein in a mammal in need of such treatment involve administration of pharmaceutical compositions that include at least one agent and / or peptide described herein in imaging and / or therapeutically effective amounts to said mammal.

[0197] Diseases, conditions, or symptoms that may benefit from treatment with compositions as described herein include, but are not limited to, endometriosis, endometriosis lesions, endometriomas, superficial endometriotic implants, deeply infiltrating endometriosis, chronic pain, central sensitization, myofascial pain, adnexal masses, infertility, dysmenorrhea, genetic predisposition, nonmenstrual pelvic-abdominal pain, dyspareunia, bowel symptoms (diarrhea, cramping, constipation), defecation pain (dyschezia), ovarian mass or tumor, painful bladder symptoms, and dysuria.

[0198] In certain embodiments, the compositions containing the agent and / or peptide(s) described herein are administered for prophylactic and / or therapeutic treatments. In certain therapeutic, imaging, and diagnostic 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 arrest at least one of the symptoms of the disease or condition. Amounts effective for this use depend on the severity and course of the disease or condition, previous therapy, the patients’ health status, weight, and response to the drugs, and the judgment of the treating physician. Imaging and therapeutically effective amounts are optionally determined by methods including, but not limited to, a dose escalation and / or dose ranging clinical trial.

[0199] In prophylactic applications, compositions containing the agents and / or peptides described herein are administered to a patient susceptible to or otherwise at risk of a particular disease, disorder or condition. Such an amount is defined to be a “prophylactically effective amount or dose.” In this use, the precise amounts also depend on the patient's state of health,weight, and the like. When used in patients, effective amounts for this use may depend on the severity and course of the disease, disorder or condition, previous therapy, the patient's health status and response to the drugs, and the judgment of the treating physician. In one aspect, prophylactic treatments include administering to a mammal, who previously experienced at least one symptom of the disease being treated and is currently in remission, a pharmaceutical composition comprising an agent and / or peptide described herein in order to prevent a return of the symptoms of the disease or condition.

[0200] In certain embodiments, wherein the patient's condition does not improve, upon the doctor's discretion, the agents and / or peptides are administered chronically, that is, for an extended period of time, including throughout the duration of the patient's life in order to ameliorate or otherwise control or limit the symptoms of the patient's disease or condition.

[0201] In certain embodiments, wherein a patient's status does improve, the dose of drug being administered is temporarily reduced or temporarily suspended for a certain length of time (i.e., a “drug holiday”). In specific 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 more than 28 days. The dose reduction during a drug holiday is, by way of example only, by 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%.

[0202] Once improvement of the patient's conditions has occurred, a maintenance dose is administered if necessary. Subsequently, in specific embodiments, the dosage or the frequency of administration, or both, is reduced, as a function of the symptoms, to a level at which the improved disease, disorder or condition is retained. In certain embodiments, however, the patient requires intermittent treatment on a long-term basis upon any recurrence of symptoms.

[0203] The amount of a given agent that corresponds to such an amount varies depending upon factors such as the particular agent and / or peptide, disease condition and its severity, the identity (e.g., weight, sex) of the subject or host in need of treatment, but nevertheless is determined according to the particular circumstances surrounding the case, including, e.g., the specific agent being administered, the route of administration, the condition being treated, and the subject or host being treated.

[0204] In general, doses employed for adult human treatment are typically in the range of from about 0.01 mg to about 5000 mg per day. In some aspects, doses employed for adult human treatment are from about 0.01 mg to about 1000 mg per day. In some embodiments, thedesired dose is conveniently presented in a single dose or in divided doses administered simultaneously or at appropriate intervals, for example as two, three, four or more sub-doses per day. In other embodiments, the composition described herein is formulated for extended 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.

[0205] In some embodiments, the daily dosages appropriate for the agent and / or peptide described herein are from about 0.01 to about 50 mg / kg per body weight. In some embodiments, the daily dosage or the amount of active in the dosage form are lower or higher than the ranges indicated herein, based on a number of variables in regard to an individual treatment regime. In various embodiments, the daily and unit dosages are altered depending on a number of variables including, but not limited to, the activity of the agent and / or peptide used, the disease or condition to be 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 practitioner.

[0206] Toxicity and efficacy of such imaging and therapeutic regimens are determined by standard pharmaceutical procedures in cell cultures or experimental animals, including, but not limited to, the determination of the LD50, IC50, EC50 and the ED50. The dose ratio between the toxic and therapeutic effects is the therapeutic index and it is expressed as the ratio between LD50 and ED50. In certain embodiments, the data obtained from cell culture assays and animal studies are used in formulating the therapeutically effective daily dosage range and / or the therapeutically effective unit dosage amount for use in mammals, including humans. In some embodiments, the daily dosage amount of the agents and / or peptides described herein lies within a range of circulating concentrations that include the ED50 with minimal toxicity. In certain embodiments, the daily dosage range and / or the unit dosage amount varies within this range depending upon the dosage form employed and the route of administration utilized.

[0207] In any of the aforementioned aspects are further embodiments in which the effective amount of the agent and / or peptide described herein is: (a) systemically administered to the mammal; and / or (b) administered orally to the mammal; and / or (c) intravenouslyadministered to the mammal; and / or (d) administered by injection to the mammal; and / or (e) administered topically to the mammal; and / or (f) administered non-systemically or locally to the mammal.

[0208] In any of the aforementioned aspects are further embodiments comprising single administrations of the effective amount of the agent and / or peptide, including further embodiments in which (i) the agent and / or peptide is administered once a day; or (ii) the agent and / or peptide is administered to the mammal multiple times over the span of one day. In additional aspects are embodiments wherein the agent and / or peptide is administered continuously over a period of time.

[0209] In any of the aforementioned aspects are further embodiments comprising multiple administrations of the effective amount of the agent and / or peptide, including further embodiments in which (i) the agent and / or peptide is administered continuously or intermittently: as in a single dose; (ii) the time between multiple administrations is every 6 hours; (iii) the agent and / or peptide is administered to the mammal every 8 hours; (iv) the agent and / or peptide is administered to the mammal every 12 hours; (v) the agent and / or peptide is administered to the mammal every 24 hours.

[0210] In any of the aforementioned aspects are further embodiments comprising the agent and / or peptide incorporated into a delivery vehicle to provide sustained delivery of the agent. In some embodiments, the 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 agent and / or is incorporated into a transdermal patch, a tape, a suppository, a vaginal suppository, vaginal tampon, vaginal ring, vaginal strip, vaginal capsule, vaginal tablet, vaginal pessary, vaginal cup, vaginal sponge, or an intrauterine device.

[0211] In further or alternative embodiments, the method comprises a drug holiday, wherein the administration of the agent and / or peptide is temporarily suspended or the dose of the agent and / or peptide being administered is temporarily reduced; at the end of the drug holiday, dosing of the agent and / or peptide is resumed. In one embodiment, the length of the drug holiday varies from 2 days to 1 year.

[0212] In certain instances, it is appropriate to administer at least one agent and / or peptide described herein in combination with one or more other therapeutic agents. Exemplary additional therapeutic agents may include, but are not limited to, hormonal therapeutic agents, including birth control, combination birth control, selective progesterone receptor antagonists, selective progesterone receptor agonists, gonadotropin-releasing hormone receptor antagonists, gonadotropin-releasing hormone receptor agonists, antiretroviral agents, antineoplastic agents, anti-inflammatories, nonsteroidal anti-inflammatories, or any combination thereof.

[0213] In one embodiment, the therapeutic effectiveness of one of the agents and / or peptides described herein is enhanced by administration of an adjuvant (i.e., by itself the adjuvant has minimal therapeutic benefit, but in combination with another therapeutic agent, the overall therapeutic benefit to the patient is enhanced). Or, in some embodiments, the benefit experienced by a patient is increased by administering one of the agents and / or peptides described herein with another agent (which also includes a therapeutic regimen) that also has therapeutic benefit.

[0214] In one specific embodiment, an agent and / or peptide described herein is coadministered with a second therapeutic agent. In some cases, the agent and / or peptide described herein, and the second therapeutic agent modulate different aspects of the disease, disorder or condition being treated, thereby providing a greater overall benefit than administration of either therapeutic agent alone. In any case, regardless of the disease, disorder or condition being treated, the overall benefit experienced by the patient may be additive of the two therapeutic agents or the patient may experience a synergistic benefit.

[0215] In certain embodiments, different effective dosages of the agents and / or peptides disclosed herein may be utilized in formulating pharmaceutical composition and / or in treatment regimens when the agents and / or peptides disclosed herein are administered in combination with one or more additional agent, such as an additional therapeutically effective drug, an adjuvant or the like, effective dosages of drugs and other agents for use in combination treatment regimens is optionally determined by means similar to those set forth hereinabove for the actives themselves. Furthermore, the methods of prevention / treatment described herein encompasses the use of metronomic dosing, i.e., providing more frequent, lower doses in order to minimize toxic side effects. In some embodiments, a combination treatment regimen encompasses treatment regimens in which administration of an agent and / or peptide described herein is initiated prior to, during, or after treatment with a second agent described herein, and continues until any time during treatment with the second agent or after termination of treatmentwith the second agent It also includes treatments in which an agent and / or peptide described herein, and the second agent being used in combination are administered simultaneously or at different times and / or at decreasing or increasing intervals during the treatment period. Combination treatment further includes periodic treatments that start and stop at various times to assist with the clinical management of the patient.

[0216] It is understood that the dosage regimen to image, diagnosis, treat, prevent, or ameliorate the condition(s) for which relief is sought can be modified in accordance with a variety of factors (e.g., the disease, disorder or condition from which the subject suffers; the age, weight, sex, diet, and medical condition of the subject). Thus, in some instances, the dosage regimen actually employed varies and, in some embodiments, deviates from the dosage regimens set forth herein.

[0217] For combination therapies described herein, dosages of the co-administered agents and / or peptides vary depending on the type of co-drug employed, on the specific drug employed, on the disease or condition being treated and so forth. In additional embodiments, when co-administered with one or more other therapeutic agents, the agent and / or peptide provided herein is administered either simultaneously with the one or more other therapeutic agents, or sequentially.

[0218] In combination therapies, the multiple therapeutic agents (one of which is one of the agents and / or peptides described herein) are administered in any order or even simultaneously. If administration is simultaneous, the multiple therapeutic agents are, by way of example only, provided in a single, unified form, or in multiple forms (e.g., as a single pill or as two separate pills).

[0219] The agents and / or peptides described herein, as well as combination therapies, are administered before, during or after the occurrence of a disease or condition, and the timing of administering the composition containing an agent and / or peptide varies. Thus, in one embodiment, the agents and / or peptides described herein are used as a prophylactic and are administered continuously to subjects with a propensity to develop conditions or diseases in order to prevent the occurrence of the disease or condition. In another embodiment, the agents and / or peptides and compositions are administered to a subject during or as soon as possible after the onset of the symptoms. In specific embodiments, an agent and / or peptide described herein is administered as soon as is practicable after the onset of a disease or condition is detected or suspected, and for a length of time necessary for the treatment of the disease. In some embodiments, the length required for treatment varies, and the treatment length is adjusted to suit the specific needs of each subject. For example, in specific embodiments, anagent and / or peptide described herein or a formulation containing the agent and / or peptide is administered for at least 2 weeks, about 1 month to about 5 years.

[0220] Peptides and compositions described herein may be co-administered with other therapeutic agents. In embodiments described herein, a circular peptide composition may be administered with a medication to treat osteoporosis including, but not limited to, alendronate, ibandronate, risedronate, zoledronic acid, denosumab, calcitonin, estrogen, raloxifene, bazodoxifene, teriparatide, abaloparatide, or any combination thereof.Embodiments

[0221] Embodiment 1. An imaging agent comprising a peptide or peptidomimetic conjugated to an imaging moiety by a linker, wherein:(a) the peptide or peptidomimetic comprises an amino acid sequence according to Formula:R-X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-Xl 2" Xi 3-X14-X15-X16, wherein:R is NH2, acetylation, stearic acid, palmitic acid, myristic acid, lauric acid, a Ci-C8hydrocarbon, a Ci-C8fatty acid, or null;Xi 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;X4is E, Q, N, or D;X5is S, alpha methyl S, K, D, Orn, T, or E;Xe is I, Chg, H, or L;X7 is L or I;X8is D, N, E, or Q;X9is D, E, K, Q, or Orn;X10 is H or methyl-H;Xu is V, alpha methyl V, Chg, L, I, or norvaline;X12 is Q, Aib, S, R, or N;X13 is R, K, citrulline, Orn, D, or E;X14 is V, I, L, or norvaline;X15 is W, Y, or F; andX16 is R, G, or null; and(b) the linker is null or the linker conjugates with the imaging moiety and the peptide or peptidomimetic.

[0222] Embodiment 2. An imaging agent comprising a peptide or peptidomimetic conjugated to an imaging moiety by a linker, wherein:(a) the peptide or peptidomimetic comprises an amino acid sequence according to Formula:X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12- X13-X14-X15-X16, wherein:Xi is M or null;X2is S, I, G, T, A, L, or null;X3is R, K or null;X4is a positively-charged amino acid, citrulline, Orn, D, E, 8-aminooctanoic acid, or an amino carboxylic acid with between 4 and 12 carbons;X5is M, Norleucine, Orn, D, E, K, H, R, K, 8-aminooctanoic acid, an amino carboxylic acid with between 4 and 12 carbons or null;X6is W, Y, F, or N-methyl A;X7is F, I, L, Chg, Cha, or Tie;Xs is L, I, or A;X9 is L, I, or A;X10 is C, S, A, Abu, C(me), or S(Bzl);X11 is F, H, A, K, E, Chg, Cng, or Orn;X12 is W, Y, A, or F; andX13 is G, GABA, or null;X14 is V, I, L, norvaline or null;X15 is W, Y, F, or null;X is R, G, or null.(b) the linker is null or the linker conjugates with the imaging moiety and the peptide or peptidomimetic.

[0223] Embodiment 3. The imaging agent of embodiment 1 or 2, wherein the imaging moiety is a fluorescent molecule, a magnetic resonance imaging (MRI) contrast agent, a radionuclide, a microbubble, a computed tomography (CT) contrast agent, or a photoacoustic contrast agent.

[0224] Embodiment 4. The imaging agent of embodiment 1 or 2, wherein the imaging moiety is a fluorophore, a fluorochrome, a dye, a pigment, a fluorescent transition metal, or a fluorescent protein.

[0225] Embodiment 5. The imaging agent of embodiment 1 or 2, wherein the imaging moiety is a paramagnetic ion complex or a superparamagnetic magnetite particle.

[0226] Embodiment 6. The imaging agent of embodiment 1 or 2, wherein the imaging moiety is a gadolinium-based contrast agent, a manganese-based contrast agent, an iron oxide contrast agent, an iron platinum contrast agent, or a ferric complex.

[0227] Embodiment 7. The imaging agent of embodiment 1 or 2, wherein the imaging moiety is a radionuclide comprising at least one selected from the group consisting of18F,64Cu,68Ga, an99mTc.

[0228] Embodiment 8 The imaging agent of embodiment 1 or 2, wherein the imaging moiety comprises a superparamagnetic iron oxide nanoparticles (SPIONs) or a DOTA-Fe(lll) complex.

[0229] Embodiment 9. The imaging agent of embodiment 1 or 2, wherein the imaging moiety is a gold nanoparticle or an iodinated compound.

[0230] Embodiment 10. The imaging agent of any one of embodiments 1-9, wherein the linker comprises one or more of amino acid, -NH(CH2)nC(O)O-, -(OCH2CH2)m-, -O-, -NH-, Ci-Ce alkylene, Ci-Ce haloalkylene, Ci-Ce alkoxy, Ci-Ce heteroalkylene, C2-Ce alkenylene, C2-Ce alkynylene, alkylamino, alkylthio, alkylcarbonyl, alkoxycarbonyl, alkylsulfonyl, C3-C10 cycloalkylene, 3- to 10-membered heterocycloalkylene, Ce-Cw aryl, or 5- to 10-membered heteroarylene, or a combination thereof, wherein each Ci-Ce alkylene, Ci-Ce haloalkylene, Ci-Ce alkoxy, Ci-Ce heteroalkylene, C2-Ce alkenylene, C2-Ce alkynylene, alkylamino, alkylthio, alkylcarbonyl, alkoxycarbonyl, alkylsulfonyl, C3-C10 cycloalkylene, 3- to 10-membered heterocycloalkylene, Ce-Cw aryl, and 5- to 10-membered heteroarylene is optionally substituted independently with 1 , 2, or 3 R9; wherein: n is independently an integer from 1 to 10; m is independently an integer from 1 to 10; andR9is independently halogen, C1-C4 alkyl, C2-C4 alkenyl, C2-C4 alkynyl, C1-C4 haloalkyl, C1-C4 alkoxy, C1-C4 haloalkoxy, C1-C4 heteroalkyl, -C(O)H, -C(O)OH, -CN, C3-C10 cycloalkyl, 3- to 10-membered heterocycloalkyl, Ce-Cw aryl, 5- to 10-membered heteroaryl, -C(O)(Ci-C4alkyl), -C(O)O(Ci-C4alkyl), -C(O)NH2, -C(O)NH(CI-C4alkyl), -C(O)N(CI-C4alkyl)2, -NH2, -NH(CI-C4alkyl), -N(CI-C4alkyl)2, -NH(C2-C4alkylene)- OH, -NH(C2-C4alkylene)-O-(Ci-C4alkyl), -OH, -O(Ci-C4alkyl), -O(Ci-C4haloalkyl), - O(C2-C4alkylene)-NH2, -O(C2-C4alkylene)-NH-(Ci-C4alkyl), -O(C2-C4alkylene)-N-(Ci- C4alkyl)2, -O(Ci-C4alkylene)-C(O)OH, -O(Ci-C4alkylene)-C(O)O-(Ci-C4alkyl), -O(C2-C4 alkenyl), -0(Ci-C4 alkylene)-(Ce-Cio aryl), -0(Ci-C4 alkylene)-(5- to 10-membered heteroaryl), 0(C6-Cio aryl), SH, S(O)2OH, -S(O)2(Ci-C4alkyl), -S(O)2NH2, - S(O)2NH(CI-C4 alkyl), or -S(O)2N(CI-C4 alkyl)2; or two R9, together with atoms to which they are attached, form a C3-C10 cycloalkyl or a 3- to 10-membered heterocycloalkyl ring.

[0231] Embodiment 11. The imaging agent of embodiment 10, wherein the linker comprise a cysteine, -NH(CH2)nC(O)-, -C(O)(CH2)nNH-, -(OCH2CH2)m-, -O-, -NH-, Ci-C6alkylene, Ci-Ce alkoxy, C-i-Ce heteroalkylene, C3-C10 cycloalkylene, or 3- to 10-membered heterocycloalkylene, or a combination thereof, wherein each C-i-Ce alkylene, Ci-Ce alkoxy, Ci-Ce heteroalkylene, C3-C10 cycloalkylene and 3- to 10-membered heterocycloalkylene is optionally substituted independently with 1 , 2, or 3 R9, wherein n, m and R9are as defined in Embodiment 36.

[0232] Embodiment 12. The imaging agent of any one of embodiments 1-11 , wherein the linker conjugates with the peptide or peptidomimetic on at least one of the amino acid from Xi to X16.

[0233] Embodiment 13. The imaging agent of any one of embodiments 1-11 , wherein the linker conjugates with an N-terminus of the peptide or peptidomimetic.

[0234] Embodiment 14. The imaging agent of embodiment 13, wherein the peptide or peptidomimetic further comprises -NH2on the C-terminus.

[0235] Embodiment 15. The imaging agent of any one of embodiments 1-12, wherein the linker conjugates with a C-terminus of the peptide or peptidomimetic.

[0236] Embodiment 16. The imaging agent of embodiment 15, wherein the peptide or peptidomimetic further comprises Ci-Cs alkyl-C(O)- on the N-terminus.

[0237] Embodiment 17. The imaging agent of embodiment 15, wherein the peptide or peptidomimetic further comprises CH3(CH2)PC(O)- on the N-terminus, wherein p is an integer from 0 to 7.

[0238] Embodiment 18. The imaging agent of embodiment 15, wherein the peptide or peptidomimetic further comprises CH3(CH2)6C(O)- on the N-terminus.

[0239] Embodiment 19. The imaging agent of any one of embodiments 1-12, wherein the peptide or peptidomimetic comprises the amino acid sequence SEQ ID NO:496 or SEQ ID NO:497.

[0240] Embodiment 20. The imaging agent of any one of embodiments 1-21 , wherein the imaging moiety is an iron oxide nanoparticle.

[0241] Embodiment 21. The imaging agent of any one of embodiments 1-21 , wherein the imaging moiety is iron chelated to 1 ,4,7,10-tetraazacyclododecane-1 ,4,7,10-tetraacetic acid (DOTA).

[0242] Embodiment 22. The imaging agent of any one of embodiments 1-21 , wherein the imaging moiety is a fluorescent molecule, MRI contrast agent, radionuclide, microbubble, CT contrast agent, or photoacoustic contrast agent

[0243] Embodiment 23. The imaging agent of any one of embodiments 1-21 , wherein the imaging moiety is a fluorophore, fluorochrome, dye, pigment, fluorescent transition metal, or fluorescent protein.

[0244] Embodiment 24. The imaging agent of any one of embodiments 1-21 , wherein the imaging moiety is a superparamagnetic iron oxide nanoparticles (SPIONs) or gadolinium complex.

[0245] Embodiment 25. The imaging agent of any one of embodiments 1-21 , wherein the imaging moiety is an18F,64Cu,68Ga, or99mTc radionuclide.

[0246] Embodiment 26. The imaging agent of any one of embodiments 1-21 , wherein the imaging moiety is a gold nanoparticle or iodinated compound.

[0247] Embodiment 27. The imaging agent of any one of embodiments 1-26, wherein the peptide or peptidomimetic is a circularized or bicyclic.

[0248] Embodiment 28. The imaging agent of embodiment 27, wherein the peptide or peptidomimetic is circularized with a Cys-Cys disulfide bond.

[0249] Embodiment 29. The imaging agent of embodiment 27, wherein the peptide or peptidomimetic is circularized with an amide bond.

[0250] Embodiment 30. The imaging agent of embodiment 29, wherein the amide bond is head-to-tail between N-terminus and C-terminus.

[0251] Embodiment 31. The imaging agent of embodiment 29, wherein the amide bond is head-to-side chain between N- terminus and an internal COOH.

[0252] Embodiment 32. The imaging agent of embodiment 29, wherein the amide bond is side chain-to-tail between an internal NTh and C-terminus.

[0253] Embodiment 33. The imaging agent of embodiment 27, wherein the amide bond is side chain-to-side chain between an internal NTh and an internal COOH.

[0254] Embodiment 34. The imaging agent of embodiment 27, wherein the peptide or peptidomimetic is circularized using hydrocarbon stapling.

[0255] Embodiment 35. The imaging agent of embodiment 27, wherein the peptide or peptidomimetic is circularized using click chemistry.

[0256] Embodiment 36. The imaging agent of any one of embodiments 1 to 35, wherein the peptide is less than 81 amino acid residues.

[0257] Embodiment 37. The imaging agent of any one of embodiments 1 to 36, wherein the peptide comprises one or more non-natural amino acids.

[0258] Embodiment 38. The imaging agent of embodiment 36, wherein the one or more non-natural amino acids are N-m ethyl amino acids.

[0259] Embodiment 39. A method for detecting endometriosis lesions in a subject, comprising:(a) administering to the subject a formulation comprising the imaging agent of any one of embodiments 1 to 38 or 60-85; and(b) imaging the subject for the presence of the imaging moiety.

[0260] Embodiment 40. The method of embodiment 39, further comprising:(c) administering to the subject a therapeutically effective amount of a second [3- catenin-binding peptide or peptidomimetic; and(d) waiting at least 1 week, and then repeating steps (a) and (b).

[0261] Embodiment 41. The method of embodiment 39 or 40, wherein the imaging in (b) further comprises creating an image and displaying the image.

[0262] Embodiment 42. The method of embodiment 41 , wherein the image is a video, still image, real-time video, or any combination thereof.

[0263] Embodiment 43. The method of embodiment 42, wherein the subject or the image is visualized or displayed during a surgical procedure conducted on the subject due to the imaging agent.

[0264] Embodiment 44. The method of embodiment 43, wherein the imaging agent is administered during or before surgery for endometriosis diagnosis, staging, and / or excision.

[0265] Embodiment 45. The method of any one of embodiments 36 to 44, wherein the formulation is administered intravaginally or intravenously.

[0266] Embodiment 46. The method of any one of embodiments 36 to 45, wherein the imaging in (b) comprises in vivo fluorescence imaging, Magnetic Resonance Imaging (MRI), Positron Emission Tomography (PET), Single Photon Emission Computed Tomography (SPECT), ultrasound imaging, Computed Tomography (CT), or photoacoustic imaging.

[0267] Embodiment 47. The method of any one of embodiments 36 to 46, wherein the subject is human.

[0268] Embodiment 48. The method of embodiment 47, wherein the imaging comprises whole body imaging.

[0269] Embodiment 49. The method of any one of embodiments 36 to 48, wherein the subject is a cultured cell line.

[0270] Embodiment 51. The method of any one of embodiments 36 to 48, wherein the subject is an ex vivo tissue extracted from an animal.

[0271] Embodiment 52. The method of any one of embodiments 36 to 51 , wherein the second p-catenin-binding peptide or peptidomimetic comprises an amino acid sequence according to Formula:R-Xi -X2-X3-X4-X5-X6-X7-X8-X9-X10-X11 -Xi 2- X-i 3-X14-X15-X16 , wherein:R is NH2, acetylation, stearic acid, palmitic acid, myristic acid, lauric acid, a Ci-Cs hydrocarbon, a Ci-Cs fatty acid, or null;Xi is M, G, beta alanine, norleucine, norvaline, or null;X2is W, N-methyl W, R, Y, F, citrulline, or K,X3is P, W, N-methyl-W, N-ethyl-W, N-methyl A, N-ethyl A, L, Pip, Aib, Y, or F;X4is E, Q, N, or D;X5 is S, alpha methyl S, K, D, Orn, T, or E;Xe is I, Chg, H, or L;X7 is L or I;X8is D, N, E, or Q;X9is D, E, K, Q, or Orn;X10 is H or methyl-H;X11 is V, alpha methyl V, Chg, L, I, or norvaline;X12 is Q, Aib, S, R, or N;X13 is R, K, citrulline, Orn, D, or E;X14 is V, I, L, or norvaline;X15 is W, Y, or F; andX16 is R, G, or null.

[0272] Embodiment 53. The method of embodiment 52, wherein the second -catenin- binding peptide or peptidomimetic has the amino acid sequence Oct-W-NMeA-Q-Orn*-ILDE*H- Chg-QRVWG-NH2(SEQ ID NO:491).

[0273] Embodiment 54. The method of any one of embodiments 36 to 51 , wherein the second [3-caten in- binding peptide or peptidomimetic comprises an amino acid sequence according to Formula:X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12- X13-X14-X15-X16,wherein:Xi is M or null;X2is S, I, G, T, A, L, or null;X3is R, K or null;X4 is a positively-charged amino acid, citrulline, Orn, D, E, 8-aminooctanoic acid, or an amino carboxylic acid with between 4 and 12 carbons;X5is M, Norleucine, Orn, D, E, K, H, R, K, 8-aminooctanoic acid, an amino carboxylic acid with between 4 and 12 carbons or null;X6is W, Y, F, or N-methyl A;X7is F, I, L, Chg, Cha, or Tie;X8is L, I, or A;X9is L, I, or A;Xw is C, S, A, Abu, C(me), or S(Bzl);X11 is F, H, A, K, E, Chg, Cng, or Orn;X12 is W, Y, A, or F;X13 is G, GABA, or null;X14 is V, I, L, norvaline or null;X15 is W, Y, F, or null; andX16 is R, G, or null.

[0274] Embodiment 55. The method of any one of embodiments 39 to 54, wherein the peptide binds to cytoplasmic b-catenin to inhibit translocation of b-catenin to a nucleus of a cell.

[0275] Embodiment 56. The method of any one of embodiments 39 to 54, wherein the peptide binds to cytoplasmic b-catenin to maintain or increase membrane-bound b-catenin.

[0276] Embodiment 57. The method of any one of embodiments 39 to 54, wherein the peptide binds to cytoplasmic b-catenin to prevent b-catenin acting as a transcription factor to oncogenes, Matrix Metalloproteinase 9 (MMP9), or Chloride C3 Channel (CIC-3).

[0277] Embodiment 58. The method of any one of embodiments 39 to 54, wherein the peptide binds to cytoplasmic b-catenin to prevent transformation, invasion, migration, fibrogenesis, or any combination thereof, of EMS cells.

[0278] Embodiment 59. The method of any one of embodiments 39 to 54, wherein the peptide binds to cytoplasmic b-catenin to prevent b-catenin from binding to estrogen receptor (ESR1).

[0279] Embodiment 60. The method of any one of embodiments 39 to 54, wherein the peptide binds to cytoplasmic b-catenin and membrane activity of b-catenin is not decreased.

[0280] Embodiment 61. The method of any one of embodiments 39 to 54, wherein the peptide binds to cytoplasmic b-catenin and b- catenin-E-cadherin binding is not decreased.

[0281] Embodiment 62. The method of any one of embodiments 39 to 54, wherein the peptide prevents oncogenic transcription factor activity.

[0282] Embodiment 63. The method of any one of embodiments 39 to 54, wherein the amount of nuclear b-catenin in a cell of the subject is decreased by at least 5%.

[0283] Embodiment 64. The method of embodiment 62, wherein the decrease in nuclear b-catenin is relative to a cell of a control subject who was not administered the therapeutically effective amount of the pharmaceutical composition.

[0284] Embodiment 65. The method of embodiment 62, wherein the decrease in nuclear b-catenin is relative to a cell of the subject taken prior to the subject developing the condition.

[0285] Embodiment 66. The method of embodiment 62, wherein the decrease in nuclear b-catenin is relative to a cell from the subject taken at a different timepoint.

[0286] Embodiment 67. The method of any one of embodiments 39 to 66, wherein the therapeutically effective amount is from about 0.01 mg to about 1000 mg.

[0287] Embodiment 68. An imaging agent according to Formula III:Label — Linker — Peptide (Formula III); wherein:(a) the Label is an imaging moiety;(b) the Peptide comprises an amino acid sequence according to Formula IV:Xi -X2-X3-X4-X5-X6-X7-X8-X9-Xi o- X11- X12- X13- Xi4- Xi5- Xi6 (Formula IV); wherein:Xi is M, G, beta alanine, norleucine, norvaline, or null;X2is W, N-methyl W, R, Y, F, citrulline, or K,X3is P, W, N-methyl-W, N-ethyl-W, N-methyl A, N-ethyl A, L, Pip, Aib, Y, or F;X4is E, Q, N, or D;X5is S, alpha methyl S, K, D, Orn, T, or E;X6is I, Chg, H, or L;X7is L or I;X8is D, N, E, or Q;X9is D, E, K, Q, or Orn;Xw is H or methyl-H;X11 is V, alpha methyl V, Chg, L I, or norvaline;X12 is Q, Aib, S, R, or N;X13 is R, K, citrulline, Orn, D, or E;X14 is V, I, L, or norvaline;X15 is W, Y, or F; andX16 is R, G, or null; and(c) the Linker is null or the Linker conjugates with the Label and the Peptide.

[0288] Embodiment 69. The imaging agent of Embodiment 68, wherein the Label is a fluorescent molecule, a magnetic resonance imaging (MRI) contrast agent, a radionuclide, a microbubble, a computed tomography (CT) contrast agent, or a photoacoustic contrast agent.

[0289] Embodiment 70. The imaging agent of Embodiment 68, wherein the Label is a fluorophore, a fluorochrome, a dye, a pigment, a fluorescent transition metal, or a fluorescent protein.

[0290] Embodiment 71 . The imaging agent of Embodiment 68, wherein the Label is a paramagnetic ion complex or a superparamagnetic magnetite particle.

[0291] Embodiment 72. The imaging agent of Embodiment 68, wherein the Label is a gadolinium-based contrast agent, a manganese-based contrast agent, an iron oxide contrast agent, an iron platinum contrast agent, or a ferric complex.

[0292] Embodiment 73. The imaging agent of Embodiment 68, wherein the Label is a radionuclide comprising at least one selected from the group consisting of18F,64Cu,68Ga, an 99m -Q

[0293] Embodiment 74. The imaging agent of Embodiment 68, wherein the Label comprises a superparamagnetic iron oxide nanoparticles (SPIONs) or a DOTA-Fe(lll) complex.

[0294] Embodiment 75. The imaging agent of Embodiment 68, wherein the Label is a gold nanoparticle or an iodinated compound.

[0295] Embodiment 76. The imaging agent of any one of Embodiments 68-75, wherein the Linker comprises one or more of amino acid, -NH(CH2)nC(O)O-, -(OCFLCFhjm-, -O-, - NH-, Ci-Ce alkylene, Ci-Ce haloalkylene, Ci-Ce alkoxy, Ci-Ce heteroalkylene, C2-C6 alkenylene, C2-C6 alkynylene, alkylamino, alkylthio, alkylcarbonyl, alkoxycarbonyl, alkylsulfonyl, C3-C10 cycloalkylene, 3- to 10-membered heterocycloalkylene, Ce-C aryl, or 5- to 10-membered heteroarylene, or a combination thereof, wherein each C1-C6 alkylene, Ci-Ce haloalkylene, Ci-Ce alkoxy, Ci-Ce heteroalkylene, C2-C6 alkenylene, C2-C6 alkynylene, alkylamino, alkylthio, alkylcarbonyl, alkoxycarbonyl, alkylsulfonyl, C3-C10 cycloalkylene, 3- to 10-membered heterocycloalkylene, C6-Cio aryl, and 5- to 10-membered heteroarylene is optionally substituted independently with 1 , 2, or 3 R9; wherein:n is independently an integer from 1 to 10; m is independently an integer from 1 to 10; andR9is independently halogen, C1-C4 alkyl, C2-C4 alkenyl, C2-C4 alkynyl, C1-C4 haloalkyl, C1-C4 alkoxy, C1-C4 haloalkoxy, C1-C4 heteroalkyl, -C(O)H, -C(O)OH, -CN, C3-C10 cycloalkyl, 3- to 10-membered heterocycloalkyl, Ce-C aryl, 5- to 10-membered heteroaryl, -C(O)(Ci-C4alkyl), -C(O)O(Ci-C4alkyl), -C(O)NH2, -C(O)NH(CI-C4alkyl), - C(O)N(CI-C4alkyl)2, -NH2, -NH(CI-C4alkyl), -N(CI-C4alkyl)2, -NH(C2-C4alkylene)-OH, -NH(C2-C4alkylene)-O-(Ci-C4alkyl), -OH, -O(Ci-C4alkyl), -O(Ci-C4haloalkyl), -O(C2- C4alkylene)-NH2, -O(C2-C4 alkylene)-NH-(Ci-C4 alkyl), -O(C2-C4 alkylene)-N-(Ci-C4 alkyl)2, -O(Ci-C4alkylene)-C(O)OH, -O(Ci-C4alkylene)-C(O)O-(Ci-C4alkyl), -O(C2-C4alkenyl), -O(Ci-C4 alkylene)-(Ce-Cio aryl), -O(Ci-C4 alkylene)-(5- to 10-membered heteroaryl), _0(C6-Cio aryl), _SH, S(O)2OH, -S(O)2(Ci-C4alkyl), -S(O)2NH2, - S(O)2NH(CI-C4 alkyl), or -S(O)2N(CI-C4 alkyl)2; or two R9, together with atoms to which they are attached, form a C3-C10 cycloalkyl or a 3- to 10-membered heterocycloalkyl ring.

[0296] Embodiment 77. The imaging agent of Embodiment 76, wherein the Linker comprise a cysteine, -NH(CH2)nC(O)-, -C(O)(CH2)nNH-, -(OCH2CH2)m-, -O-, -NH-, Ci-C6alkylene, Ci-Ce alkoxy, Ci-Ce heteroalkylene, C3-C10 cycloalkylene, or 3- to 10-membered heterocycloalkylene, or a combination thereof, wherein each Ci-Ce alkylene, Ci-Ce alkoxy, Ci-Ce heteroalkylene, C3-C10 cycloalkylene and 3- to 10-membered heterocycloalkylene is optionally substituted independently with 1 , 2, or 3 R9, wherein n, m and R9are as defined in Embodiment 36.

[0297] Embodiment 78. The imaging agent of any one of Embodiment 68-77, wherein the Linker conjugates with the Peptide on at least one of the amino acid from Xi to X16.

[0298] Embodiment 79. The imaging agent of any one of Embodiment 68-77, wherein the Linker conjugates with an N-terminus of the Peptide.

[0299] Embodiment 80. The imaging agent of Embodiment 79, wherein the Peptide further comprises -NH2on the C-terminus.

[0300] Embodiment 81. The imaging agent of any one of Embodiment 68-77, wherein the Linker conjugates with a C-terminus of the Peptide.

[0301] Embodiment 82. The imaging agent of Embodiment 80, wherein the Peptide further comprises Ci-Cs alkyl-C(O)- on the N-terminus.

[0302] Embodiment 83. The imaging agent of Embodiment 80, wherein the Peptide further comprises CH3(CH2)PC(O)- on the N-terminus, wherein p is an integer from 0 to 7.

[0303] Embodiment 84. The imaging agent of Embodiment 80, wherein the Peptide further comprises CH3(CH2)6C(O)- on the N-terminus.

[0304] Embodiment 85. The imaging agent of any one of Embodiments 68-84, wherein the amino acid sequence is circularized with an amide bond, and wherein the amide bond is (1) head-to-tail between N-terminus and C-terminus, (2) head-to-side chain between N-terminus and an internal COOH, (3) side chain-to-tail between an internal NH2 and C-terminus, or (4) side chain-to-side chain between an internal NH2 and an internal COOH, and wherein the amide bond is linked to amino acids that are not connected with either the Linker or the Label.

[0305] A number of embodiments of the disclosure have been described. Nevertheless, it will be understood that various modifications may be made without departing from the spirit and scope of the disclosure. Accordingly, other embodiments are within the scope of the following claims. EXAMPLESExample 1 : Peptides conjugated to imaging moieties can be used as imaging agents to visualize disease in vivo as well as API / drug product characteristics

[0306] The purpose of the presented series of figures is to showcase the potential of peptide agents in advanced biomedical applications. In FIG. 1, an example related to endometriosis, compounds 492, 494, and 493 were used in animals 2-4, respectively, to show that the endometriosis lesions in the animals can be visualized with these imaging and diagnostic agents as valuable tools for imaging subjects to detect the presence of diseased tissue. The same mice sent for necropsy show ex vivo analysis similar to its live in vivo imaging.

[0307] C57BI / 6 albino mice were imaged using spectral imaging (Living Image 4.7.3, MS Spectrum, Perkin Elmer). Mice were put to sleep using isoflurane following standard procedure. Adaptive fluorescence background subtraction was used as well as spectral unmixing, epi-illumination for Cy7 with spectral emission scan from 757 to 781 nm. Each time point was normalized by setting the threshold for endometriosis mice and using that for healthy mice.

[0308] These agents can also be utilized to evaluate key characteristics of the Active Pharmaceutical Ingredient (API) or drug product. FIG. 2 shows the bio distribution of compound 491 by use of labelled peptide compound 492 which reveals the specificity of these agents, with little to no detectible agent in other organ systems. This is also supported by FIG. 15 which shows that compound 492 is only detected in endometriosis tissue and not detected in the healthy uterus tissue. FIG. 3 shows vaginal administration of compound 494 and detection atthe site of the lesion. This has been successfully shown with other labelled peptides as well as other routes of administration including but not limited to SQ, IV and PO dosing as viable options which all have different pharmacokinetic profiles. In vitro experiment of FIG. 4 confirms the visual observations in FIG. 3 in that peptides are permeable through vaginal tissue and that administration of peptides vaginally takes approximately 3 hours to cross the human vaginal tissue. This is consistent with the imaging performed via animal model with the disclosed imaging agent and confirms the accuracy of the imaging agents.

[0309] The agents and peptide described herein have shown promise in monitoring disease progression or gauging the effectiveness of therapeutic treatments. The ability to administer these agents through various routes, as described in FIG. 3 and FIG. 4, offers flexibility in the delivery of these imaging, diagnostic and / or therapeutic agents. As demonstrated in FIG. 6, time points were taken between 20 min and 72 hours and imaging was seen up to 48 hours in endometriosis subjects post blood clearance and was not visualized in subjects without endometriosis post blood clearance. In FIG. 7, the peptides were be used to track disease progression and / or lesion with treatment, thereby providing a quantitative assessment of treatment efficacy and disease burden over time. FIG. 7 shows animals dosed with 2 SQ administrations of compound 492 weeks apart and the resulting imaging, diagnostic, and efficacy effect of the agent. The final necropsy report of ex vivo lesion analysis was consistent with live visualization with agent administration. Tracking lesion regression within the same mouse after 2 SQ doses administered weeks apart showing that daily dosing is not necessary for efficacy.Example 2: Identifying Efficacious Dose and Dosing Regime

[0310] The purpose of this example, as illustrated in FIG. 8 through FIG. 10, is to identify an approximation of the efficacious dose and determine the optimal dosing regimen. FIG. 8 through 21 utilize compounds 491 or 492 as examples but have also been repeated and confirmed with many of the disclosed compounds.

[0311] FIG. 8 shows the resulting lesion weight of endometriosis mice that were administered compound 491 daily for 3 (left graph) or 6 weeks (right graph). Therapeutic agent improves with longer treatment durations, especially when administered in lower doses. This evidence provides valuable guidance for optimizing treatment schedules to achieve maximum therapeutic outcomes, particularly when utilizing lower dosage levels.

[0312] FIG. 9 shows into the pharmacokinetic exposure of compound 491 at 1mg / kg in mice as analyzed in serum. When compared with data from FIG. 6 to FIG. 8, the pharmacodynamic effects of compound 491 and sustained presence specific to the lesionextend beyond its pharmacokinetic presence in serum, indicating the agent’s sustained imaging, diagnostic and therapeutic potential.

[0313] FIG. 10 supports this finding by using a cell proliferation assay. A single dose of SEQ ID NO: 491 exerts pharmacodynamic effects that last up to five days post-treatment, outlasting its serum pharmacokinetics. The proliferation assay was used to determine the pharmacodynamic effects of compound 491 on 12Z human endometriosis cells. A 96 well plate was treated with an extracellular matrix for an hour and then seeded 12Z human endometriosis cells into the wells at a concentration of 5000 cells per 50 pL of assay media. After an overnight incubation at 37°C, 5% CO2, the media was aspirated out and fresh Prigrow III medium supplemented with 10% FBS and 1% PS containing different concentrations of the peptide of interest was added to each well. The cells were treated with the peptide for just one day, after which we added fresh media to each well every day for the next four days. To determine the effects of the peptide, the media was mixed with Presto Blue, which turned pink within 4-6 hours. The fluorescence of each well was measured at 540nm and 590nm using a Biotek HTX microplate reader. The IC50 and graphs was calculated using Graph Pad Prism 9.0, providing a quantitative measure of the peptide's effect on cell proliferation. One dose of compound 491 has lasting pharmacodynamic effects (5 days are shown post treatment). This demonstrates that increased dosing frequency can potentially enhance outcomes but is not necessary for efficacy.Example 3: In vitro characterization of agents

[0314] The purpose of these study examples is to demonstrate that the diagnostic peptides remain cell permeable even when conjugated to an imaging moiety and are able to have a visualized signal without quenching from conjugation, intracellular location, or peptide binding to p-catenin target. Furthermore, the imaging moiety does not interfere with the diagnostic peptides' ability to localize in the cytoplasm or disrupt its cytoplasm p-catenin binding, translation from cytoplasm to inner membrane, or its p-catenin membrane binding. Cy7-labeled compound 491 (compound 492) is able to engage target p-catenin, can successfully translocate with target bound to the inner membrane, is able to be imaged with the moiety conjugated to the peptide without signal disruption from its conjugation the peptide or target binding, has quick intracellular uptake, and is retained in the disease tissue longer than in serum. These set of in vitro data support the in vivo imaging studies and pharmacodynamic investigations shown in the other figures.

[0315] As shown in FIG. 11 and FIG. 12, compound 492 binds to target p-catenin directly within the cell, it is cell-permeable, and it binds to target in the cytoplasm of cell. Thisbinding does not occur in the nucleus and does not appear to disrupt membrane binding. To determine whether compound 491 is cell permeable, the cellular localization of compound 492, the binding with its binding partner (P-catenin) and whether it could be visualized with a fluorophore, human endometriosis 12z cells were incubated with compound 492 for 1h. Membrane fraction, cytoplasmic fraction and nuclear fraction were extracted. BCA analysis was done to determine the concentration of protein. Equal amount of protein from all fractions along with compound 492 were run on Native PAGE gel. These results show that both compound 492and the p-catenin / compound 492 binding complex are able to be visualized and separated on the NATIVE gel, and that compound 492 was present in the membrane and cytoplasm fraction, but not nuclear fraction confirming mechanism of action.

[0316] To determine at what time point compound 491 binds to p-catenin in cytoplasm, human endometriosis 12z cells were dosed once with 0.5pM compound 492 for different timepoints i.e., 1h, 3h, 5h, 7h, 1day, 2days and 3days. After the dosage, cells were washed with PBS buffer and cytoplasmic fraction was extracted. Native gel was run and imaged. The results in FIG. 14 suggest that compound 492 bind to p-catenin within 1 hr. of incubation, increases in signal intensity for 24h, and exhibits a stable signal for several days. These findings are in according to previous experiments: 1) FP experiment, where binding occurs after 1 hour and remains until at least after 24 hours (data not shown), and (2) imagining experiments in rodent model lesions where binding remains in lesions at least 48 hours (FIG. 6) whereas rodents without endometriosis show no signs of the imaging agent post blood clearance (FIG. 6). Additionally, this also is consistent with proliferation results which showed that just one dose of compound 491 provides lasting pharmacodynamic effects of decreased cell proliferation.

[0317] The results in FIG. 12 show that compound 492 bind to p-catenin with 1 hr. incubation and signal is stable up to at least 3 days. These findings are in according to our previous experiments. 1) FP experiment, where binding occurs after 1 hour and remains until at least after 24 hours and (2) imagining experiments in rodent model lesions where binding remains in lesions beyond blood clearance. This is consistent with animal imaging results in mice which shows that one dose of compound 492 remains specifically in diseased lesion.

[0318] To visualize the effect on compound 491 on p-catenin in cytoplasmic fraction human endometriosis (12z cells) were dosed with 0.5pM compound 492 and timepoints i.e., 1 h, 3h, 5h, 7h, 1day, 2days and 3days post-treatment was analyzed. After the dosage, cells were washed with PBS buffer and cytoplasmic fraction was extracted. Native gel was run, which was imaged using Cy7. Gel was then transferred to perform western blot using nitrocellulose membrane and was probed with beta catenin antibody. Western blot was imaged usingchemiluminescence. The results in FIG. 13 suggest that [3-catenin protein starts decreasing after compound 492 binding occurs in cytoplasm.

[0319] To visualize the effect on compound 491 on p-catenin in membrane fraction, human endometriosis 12z cells were dosed once with 0.5pM compound 492 and different timepoints i.e., 1 h, 3h, 5h, 7h, 1day, 2day post-treatment was analyzed. After the dosage, cells were washed with PBS buffer and non-cytoplasmic / non-nuclear fraction was extracted. Native gel was run, which was imaged using Cy7. Gel was then transferred to perform western blot using nitrocellulose membrane and was probed with beta catenin antibody. Western blot was imaged using chemiluminescence. The results in FIG. 13 suggest that [3-catenin protein remains stable in the membrane fraction.

[0320] These observations suggest that peptides can be inherently flexible, in that its interaction with one of the (TCF4) binding site can influence allosteric confirmation of the other (BCL9) binding sites even though only one of the binding sites is physically interacting with compound 491. This flexibility can aid in the relocalizing and / or supportive binding of [3-catenin from cytoplasm to membrane, allowing for its Wnt4 rescue activity and confirming mechanism of action.Example 4: Species homology

[0321] Also herein, it is shown that the peptides can have high homology across species (in this example, compound 491 in excess of 95% across various animal species), and thus can be applicable in all animal and mammalian species for imaging, diagnostic, and therapeutic interventions. According to the data presented in Table 3, the binding characteristics of compound 491 to [3-catenin exhibit substantial similarity across various mammalian species. Therefore, compound 492and other imaging agents utilizing compound 491 can be effectively employed in animal studies to visualize the progression or elimination of endometriosis disease using therapeutic agents or other chemical compounds. This offers a big advantage over the use of animal cohorts and evaluate treatment efficacy for many reason, including but not limiting to ability to view efficacy real time while not needing to sacrifice the animal, seeing the efficacy of the therapeutic within the same animals, which is very useful in a heterogenous disease such as endometriosis.

[0322] The binding data across all mammals are shown in Table 3. As shown in Table 3, binding of compound 491 among mouse, rat, dog, and human [3-catenin is nearly identical.Example 5: Allosteric confirmation of p-catenin target and flexibility in ring yields wnt4 activity for both efficacy and safety

[0323] The purpose of this example is to show the allosteric conformational change in p- catenin, where binding at one site induces a structural transformation throughout the entire molecule, allowing for its translocation from cytoplasm to its interaction with the cellular membrane, p-catenin is inherently known to have several structures which allow it to either be in the membrane or cytoplasm. Previously it was hypothesized that TCF4 and BCL9 binding sites could not be simultaneously affected by binding to a small molecule because the binding sites were on opposite sides of a large protein. However, Table 4 describes that binding of agents and / or peptides to p-catenin can have an affect across both sites, similarly, indicating the inherent full protein flexibility of [3-catenin due to its conformational adaptation. This flexibility can enable cytoplasmic [3-catenin to adopt a conformation that facilitates membrane localization, similar to the action of Wnt4 agonists. Consequently, this demonstrates the ability to bind both cytoplasmic and membrane-bound forms of [3-catenin due to its flexibility. The ability to either relocalize and / or support [3-catenin binding to the membrane allows for the efficacy of the agent, while allowing for [3-catenin to remain in the membrane allows for both the safety of the peptide and mechanism of action. This activity is shown in FIG. 15 where the cellular localization of peptide which is majority at the membrane after equilibrium has been reached, which suggests Wnt4 activity of the peptides and FIG. 16 quantitates the observations in FIG 15.

[0324] Binding of and agents to one site in [3-catenin may promote allosteric Binding of and agents to one site in [3-catenin may promote allosteric confirmational change, affecting both BCL9 and TCF4 binding sites as shown in Table 4.Example 6: On target effect and downstream targets

[0325] This example shows the on-target effects and mechanism of action of agents, specifically compound 491 , and its impact on the [3-catenin signaling pathway resulting in imaging, diagnostic, and / or therapeutic potential of diseases driven by p-catenin dysregulation. As endometriosis regression was observed in mice following the 3 or 6 week treatment of compound 491 (FIG. 8), on-target changes were examined in resulting excised endometriotic lesions. FIG. 17-21 show the impact of the administration of compound 491 in animals on the extracted endometriosis lesions post-treatment. There is a decrease cell epithelial proliferation (FIG. 17), increase in cell epithelial apoptosis (FIG. 18), increase in activated macrophage recruitment (FIG. 19), decrease in downstream targeted p-catenin RNA (FIG. 20), and decrease in downstream targeted p-catenin proteins (FIG. 21). These figures support the mechanism of action and on target activity of these peptides and these sets of experiment have been repeated with many of the disclosed compounds.

[0326] FIG. 22 also shows that a labeled peptide targeting downstream target protein MMP2 can be used to image diseased tissue (in this example, endometriosis) in an endometriosis mouse model.Example 7: Peptide pKa values and pH differences in endometriotic cells drive peptide specificity.

[0327] Peptide specificity to diseased cells has been documented. This is supported by the understanding that unhealthy cells exhibit a different intracellular pH (pHi) compared to healthy cells, allowing for increased permeability in pathologic cells and reduced permeability in healthy cells. We tested endometriosis and healthy cells via intracellular pH indicator dyes (Invitrogen™, cat. no. P35372) and discovered that the pH of healthy cells was 7.2 and pH of endometriosis cells was 4.5. To assess if compound 491 undergoes changes in membrane permeability with pH variations, PAMPA was performed across pH 3 to pH 10 according to published literature. We confirmed that compound 491 permeability differed significantly from high to low permeably depending on pH. As two examples, pKa distribution of compound 491 and compound 430 pKa showed a pKa value 6.7 and 5.2, respectively; both values falling between the pH of endometriosis cells (4.5) and healthy cells (7.2). Together, the data suggest that these compounds can observe drastic changes in cellular permeability based on protonation status in response to different pH environments, driving selectivity in endometriosis cells.Example 8: Peptides disclosed herein do not demonstrate off-target activity.

[0328] Compound 491 was tested across Panlabs SAFETYScan, a safety panel with 78 targets used for safety evaluation in drug discovery to predict clinical adverse events. Compound 491 did not show any off-target activity, including transporters for cardiac arrhythmia assessment — hERG, CaV, and NaV., even at highest concentrations tested.Example 9: Peptides disclosed herein do not affect HPG axis, clinical chemistry, or hematology.

[0329] To visualize potential off-target effects on reproductive organ systems, uterus and ovary, and peritoneal wall were visualized by H&E after compound 491 treatment. No toxicity or necrotic off-target effects were observed in these organs (uterus and ovary; Fig. 23). In addition, compound 491 did not alter uterine cyclicity / HPG-axis or uterine weight at necropsy (data not shown). Standard veterinary clinical chemistry parameters (Table 5) and hematology (data not shown) findings were within normal limits after compound 491 treatment. Further toxicology studies confirmed no observed adverse effects at maximum feasible dose (>2000x expected clinical dose exposure).

[0330] Together, the preliminary data lay a robust groundwork, demonstrating that peptides from our library can serve as effective diagnostic agents because the peptides (1) bind directly to -catenin even when conjugated to an imaging moiety, (2) are cell permeable even when conjugated to an imaging moiety, (3) have detectable signals which are not dampened by conjugation to peptide, (4) exhibit a sustained half-life in lesions following blood clearance, (5) are not absorbed by healthy cells, (6) accurately detect lesions during in vivo live imaging compared to ex vivo necropsy reports, (7) can quantify disease burden and therapeutic efficacy, (8) demonstrate pH-selective targeting of diseased cells based on pKa properties, (9) show no toxicities or off-target activities, and (10) are non-hormonal. Therefore, our central hypothesis isthat a novel, non-hormonal peptide specific for endometriosis cells conjugated with iron-imaging moieties will serve as a safe and targeted MRI imaging agent. The imaging agent is expected to detect endometriosis reliably, specifically, and sensitively through non-invasive methods.Example 10: Pharmacokinetics (PK) confirm peptide blood clearance.

[0331] PK studies were performed in CD1 healthy mice to profile absorption, distribution, and excretion of compound 491 in blood plasma (Table 6) via LC / MS. Compound 491 was detected for up to 24 h post-administration at which point it was cleared from the bloodstream, similar to imaging results. Subjects with endometriosis retained the compound in their lesions specifically which was still able to be visualized after plasma clearance compared to subjects without endometriosis which had no detectible amounts of compound after the compound was cleared from the bloodstream (FIG. 1 and 6). Collectively, these data confirm that the imaging potential of image labeled compound 491 for visualizing endometriosis lesions extends beyond its plasma presence.Example 11 : Evaluation of Pharmacokinetics, Therapeutic Activity, and Target Binding in Diagnostic Compounds

[0332] We tested several compounds in the library that bind specifically to B-catenin and looked at the relation to therapeutic pharmacodynamics and pharmacokinetics in plasma and lesion. The purpose of these experiments was to investigate if compounds are required to be highly therapeutically active to exhibit diagnostic effects.

[0333] Reporter assays and target binding were performed in accordance to Table 4 protocols. Protein binding was measured in 5 human plasma (10 pM final) and incubated with rotation at 37 C for 4 hr using the Rapid Equilibrium Dialysis device (Thermo Scientific™). Samples from both sides of the dialysis device were be prepped for LC-MS / MS analysis with propranolol hydrochloride will be used as a positive control. The percentage protein binding was calculated as follows: Protein binding (%) = (Areap - Areab / Areap) x 100. Plasma stability was analyzed by pre-incubating human plasma (990 uL) in water bath at 37°C and 75 rpm for 5 min. 10 pL compound was added. 50 pl aliquots taken in duplicate at 0, 0.5, 2, 3.5, 4, 4.5, and 5minutes to a 1.5 mL tube containing 150 pl of warfarin (200ng / mL). The tubes were incubated in water bath at 37°C and 75 rpm for up to 120 min. The samples were then centrifuged at 17000xg for 10 min at 4°C. 100 pl of supernatant was transferred to a 96-well plate containing 100 pl of water, vortexed, and then spun down at 3750 rpm for 10 minutes. Pharmacokinetic analysis was performed by administering 1mg / kg of compound to CD1 mice and Blood collection will be at the following time points: pre-dose and 0.083, 0.25, 0.5, 1 , 2, 4, 8, 12, and 24 h post-dose. After centrifugation, the collected plasma will be analyzed via LC-MS / MS to determine t1 / 2 and concentrations of the peptide in ng / ml based on a standard curve. Pharmacodynamic effect of each peptide was analyzed by administering a low dose of 0.325mg / kg or 1mg / kg / d of peptide IP for 3 weeks. Results are shown in Table 8.

[0334] A critical attribute was the binding affinity of the peptides to the target. All compounds exhibiting some degree of plasma stability, however, critically, compounds were retained specifically in the diseased lesions longer than its presence in the bloodstream, allowing for disease imaging to occur post plasma clearance. Compounds with longer half-lives showed low to undetectable plasma levels at 24 hours, yet significant retention within lesions, indicative of an effective diagnostic compound. Plasma degradation varied and was inversely proportional to protein binding, suggesting, without bound by theory, that protein or target binding within the cell shields the compound from degradation. Without bound by theory, once compounds translocated inside the cell, the compounds may be shielded from plasma proteases either by intracellular localization or binding to intracellular targets, suggesting that target binding likely contributes to reduced degradation in the lesions. Regardless of strength of therapeutic potency, retention within the lesion surpassed plasma levels, affirming their potential as diagnostic candidates. Without bound by theory, quick clearance from the bloodstream could contribute to diminished therapeutic efficacy, yet, still allows for effective diagnostic performance. Therefore, compounds with or without strong therapeutic efficacy are still strong diagnostic candidates due to their clearance from the bloodstream and specific uptake by disease cells.Example 12: Conjugate diagnostic peptides to iron-based moieties

[0335] Peptides were conjugated to functionalized iron oxide nanoparticles to enable MRI detection. For this example, iron oxide nanoparticles <100nm are used because these small particles have good potential for cellular targeting and are below the 100 nm range where immune system recognition is less likely. Additionally, smaller nanoparticles have observed Enhanced Permeability Effect (EPR) because unhealthy cells are surrounded by blood vessel endothelial cells with enhanced permeability through leaky junctions.

[0336] The example here describes the conjugation of peptides that are performed with methods that include either functionalized iron oxide nanoparticles or [lron]DOTA, enabling their visualization through MRI imaging. In FIG 24A, for the iron nanoparticles, functionalized iron oxide nanoparticles and conjugation kits were sourced from Ocean Nanotech (San Diego, CA). Hydrodynamic size, PDI and zeta potential are used to confirm using a Malvern Zetasizer (Malvern Panalytical, Malvern, Worcestershire, United Kingdom). To enable conjugation to peptides containing a cysteine at N-terminal with amine functionalized iron oxide nanoparticles, the nanoparticles are reacted with Sulfo-SMCC (succinimidyl 4-(N-maleimidomethyl) cyclohexane- 1 -carboxylate) to facilitate the formation of stable amide bond. The free thiol group present on the peptides will react with the intermediate functionalized iron oxide nanoparticles to generate peptide-iron oxide conjugate. See FIG 24B. (see FIG 24B). To enable conjugation to peptides via carboxyl-functionalized iron oxide nanoparticles, the carboxyl groups on the nanoparticles are activated using a carbodiimide chemistry involving EDC (1 -ethyl-3-(3- dimethylaminopropyl) carbodiimide) and NHS (N-hydroxysuccinimide) to form stable amide bonds with amine groups at the N-terminal of the peptides For [lron]DOTA conjugated peptides, the DOTA moiety is first linked to the peptide through solid-phase peptide synthesis (SPPS), targeting either the N-terminus, C-terminus, or a specific side chain, ensuring the functionality and stability of the final complex. Next, iron ions are introduced to the DOTA- peptide conjugate to facilitate the efficient chelation of iron within the DOTA cage. The resulting iron conjugated peptides undergoes purification using high-performance liquid chromatography (HPLC) to remove any unreacted components and byproducts, followed by a thorough characterization of the complex.Example 13: Additional examples of imaging.Peptides were synthesized with fluorophores to enable imaging as disclosed herein (Table 7):TH Docket No. 170502-2010TH Docket No. 170502-2010

[0337] Unless defined otherwise, all technical and scientific terms used herein have the same meanings as commonly understood by one of skill in the art to which the disclosed disclosure belongs. Publications cited herein and the materials for which they are cited are specifically incorporated by reference.

[0338] Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the disclosure described herein. Such equivalents are intended to be encompassed by the following claims.

Claims

CLAIMS1. An imaging agent comprising a peptide or peptidomimetic conjugated to an imaging moiety by a linker, wherein:(a1) the peptide or peptidomimetic comprises an amino acid sequence according to Formula I:R-Xi-X2-X3-X4-X5-X6-X7-X8-X9-Xw-Xii-Xi2- XI3-XI4-XI5-XI6(Formula I), wherein:R is NH2, acetylation, stearic acid, palmitic acid, myristic acid, lauric acid, a Ci-Cs hydrocarbon, a Ci-Cs fatty acid, or null;Xi is M, G, beta alanine, norleucine, norvaline, or null;X2is W, N-methyl W, R, Y, F, citrulline, or K,X3is 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;X5is S, alpha methyl S, K, D, Orn, T, or E;X6is I, Chg, H, or L;X7is L or I;X8is D, N, E, or Q;X9is D, E, K, Q, or Orn;X is H or methyl-H;X11 is V, alpha methyl V, Chg, L, I, or norvaline;X12 is Q, Aib, S, R, or N;X13 is R, K, citrulline, Orn, D, or E;X14 is V, I, L, or norvaline;X15 is W, Y, or F; andX16 is R, G, or null; or(a2) the peptide or peptidomimetic comprises an amino acid sequence according to Formula II:X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12- Xi3-Xi4-Xi5-Xi6 (Formula II), wherein: Xi is M or null;X2is S, I, G, T, A, L, or null;X3is R, K or null;X4is a positively-charged amino acid, citrulline, Orn, D, E, 8-aminooctanoic acid, or an amino carboxylic acid with between 4 and 12 carbons;Xs is M, Norleucine, Orn, D, E, K, H, R, K, 8-aminooctanoic acid, an amino carboxylic acid with between 4 and 12 carbons or null;X6is W, Y, F, or N-methyl A;X7is F, I, L, Chg, Cha, or Tie;Xs is L, I, or A;X9is L, I, or A;Xw is C, S, A, Abu, C(me), or S(Bzl);Xn is F, H, A, K, E, Chg, Cng, or Orn;X12 is W, Y, A, or F; andX13 is G, GABA, or null;X14 is V, I, L, norvaline or null;X15 is W, Y, F, or null; andX is R, G, or null; and(b) the linker is null or the linker conjugates with the imaging moiety and the peptide or peptidomimetic.

2. The imaging agent of claim 1 , wherein the imaging moiety is a fluorescent molecule, a magnetic resonance imaging (MRI) contrast agent, a radionuclide, a microbubble, a computed tomography (CT) contrast agent, or a photoacoustic contrast agent.

3. The imaging agent of claim 2, wherein the imaging moiety is a fluorophore, a fluorochrome, a dye, a pigment, a fluorescent transition metal, or a fluorescent protein.

4. The imaging agent of claim 2, wherein the imaging moiety is a paramagnetic ion complex or a superparamagnetic magnetite particle.

5. The imaging agent of claim 2, wherein the imaging moiety is a gadolinium-based contrast agent, a manganese-based contrast agent, an iron oxide contrast agent, an iron platinum contrast agent, or a ferric complex.

6. The imaging agent of claim 2, wherein the imaging moiety is a radionuclide comprising at least one selected from the group consisting of18F,64Cu,68Ga, an99mTc.

7. The imaging agent of claim 2, wherein the imaging moiety comprises a superparamagnetic iron oxide nanoparticles (SPIONs) or a DOTA-Fe(lll) complex.

8. The imaging agent of claim 2, wherein the imaging moiety is a gold nanoparticle or an iodinated compound.

9. The imaging agent of claim 1 , wherein the linker comprises one or more of amino acid, - NH(CH2)nC(O)O-, -(OCH2CH2)m- -O-, -NH-, Ci-C6alkylene, Ci-C6haloalkylene, Ci-C6alkoxy, Ci-Ce heteroalkylene, C2-Ce alkenylene, C2-Ce alkynylene, alkylamino, alkylthio,alkylcarbonyl, alkoxycarbonyl, alkylsulfonyl, C3-C10 cycloalkylene, 3- to 10-membered heterocycloalkylene, Ce-Cw aryl, or 5- to 10-membered heteroarylene, or a combination thereof, wherein each Ci-Ce alkylene, Ci-Ce haloalkylene, Ci-Ce alkoxy, Ci-Ce heteroalkylene, C2-C6 alkenylene, C2-C6 alkynylene, alkylamino, alkylthio, alkylcarbonyl, alkoxycarbonyl, alkylsulfonyl, C3-C10 cycloalkylene, 3- to 10-membered heterocycloalkylene, Ce-C aryl, and 5- to 10- membered heteroarylene is optionally substituted independently with 1, 2, or 3 R9; wherein: n is independently an integer from 1 to 10; m is independently an integer from 1 to 10; andR9is independently halogen, C1-C4 alkyl, C2-C4 alkenyl, C2-C4 alkynyl, C1-C4 haloalkyl, C1-C4 alkoxy, C1-C4 haloalkoxy, C1-C4 heteroalkyl, -C(O)H, -C(O)OH, -CN, Cs-Cw cycloalkyl, 3- to 10-membered heterocycloalkyl, Ce-Cw aryl, 5- to 10-membered heteroaryl, -C(O)(Ci-C4alkyl), -C(O)O(Ci-C4alkyl), -C(O)NH2, -C(O)NH(CI-C4alkyl), -C(O)N(CI-C4alkyl)2, -NH2, -NH(CI-C4alkyl), -N(CI-C4alkyl)2, -NH(C2-C4alkylene)- OH, -NH(C2-C4alkylene)-O-(Ci-C4alkyl), -OH, -O(Ci-C4alkyl), -O(Ci-C4haloalkyl), - O(C2-C4alkylene)-NH2, -O(C2-C4alkylene)-NH-(Ci-C4alkyl), -O(C2-C4alkylene)-N-(Ci- C4alkyl)2, -O(Ci-C4alkylene)-C(O)OH, -O(Ci-C4alkylene)-C(O)O-(Ci-C4alkyl), -O(C2- C4 alkenyl), -O(Ci-C4 alkylene)-(Ce-Cw aryl), -O(Ci-C4 alkylene)-(5- to 10-membered heteroaryl), _0(C6-Cw aryl), _SH, S(O)2OH, -S(O)2(Ci-C4alkyl), -S(O)2NH2, - S(O)2NH(CI-C4 alkyl), or -S(O)2N(CI-C4 alkyl)2; or two R9, together with atoms to which they are attached, form a Cs-Cw cycloalkyl or a 3- to 10-membered heterocycloalkyl ring.

10. The imaging agent of claim 9, wherein the linker comprise a cysteine, -NH(CH2)nC(O)-, -C(O)(CH2)nNH-, -(OCH2CH2)m-, -O-, -NH-, CrC6alkylene, Ci-C6alkoxy, Ci-C6heteroalkylene, C3-Cw cycloalkylene, or 3- to 10-membered heterocycloalkylene, or a combination thereof, wherein each Ci-Ce alkylene, Ci-Ce alkoxy, Ci-Ce heteroalkylene, Cs-Cw cycloalkylene and 3- to 10-membered heterocycloalkylene is optionally substituted independently with 1 , 2, or 3 R9, wherein n, m and R9are as defined in claim 36.

11. The imaging agent of claim 1 , wherein the linker conjugates with the peptide or peptidomimetic on at least one of the amino acid from Xi to X .

12. The imaging agent of claim 1 , wherein the linker conjugates with an N-terminus of the peptide or peptidomimetic.

13. The imaging agent of claim 12, wherein the peptide or peptidomimetic further comprises -NH2 on the C-terminus.

14. The imaging agent of claim 1 , wherein the linker conjugates with a C-terminus of the peptide or peptidomimetic.

15. The imaging agent of claim 14, wherein the peptide or peptidomimetic further comprises Ci-Cs alkyl-C(O)- on the N-terminus.

16. The imaging agent of claim 14, wherein the peptide or peptidomimetic further comprises CH3(CH2)PC(O)- on the N-terminus, wherein p is an integer from 0 to 7.

17. The imaging agent of claim 14, wherein the peptide or peptidomimetic further comprises CH3(CH2)6C(O)- on the N-terminus.

18. The imaging agent of claim 1 , wherein the peptide or peptidomimetic comprises the amino acid sequence SEQ ID NO:496 or SEQ ID NO:497.

19. The imaging agent of claim 18, wherein the imaging moiety is an iron oxide nanoparticle.

20. The imaging agent of claim 18, wherein the imaging moiety is iron chelated to 1 ,4,7,10- tetraazacyclododecane-1 ,4,7,10-tetraacetic acid (DOTA).

21. The imaging agent of claim 1 , wherein the peptide or peptidomimetic is a circularized or bicyclic.

22. The imaging agent of claim 21 , wherein the peptide or peptidomimetic is circularized with a Cys-Cys disulfide bond, an amide bond, using hydrocarbon stapling, or using click chemistry.

23. The imaging agent of claim 22, wherein the amide bond is head-to-tail between N- terminus and C-terminus, wherein the amide bond is head-to-side chain between N-terminus and an internal COOH, wherein the amide bond is side chain-to-tail between an internal NTh and C-terminus, wherein the amide bond is side chain-to-side chain between an internal NTh and an internal COOH, or any combination thereof..

24. A method for detecting endometriosis lesions in a subject, comprising:(a) administering to the subject a formulation comprising the imaging agent of any one of claims 1 to 23; and(b) imaging the subject for the presence of the imaging agent.

25. The method of claim 24, further comprising:(c) administering to the subject a second -catenin-binding peptide or peptidomimetic; and(d) waiting at least 1 week, and then repeating steps (a) and (b).

26. The method of claim 24, wherein the imaging in (b) further comprises creating an image and displaying the image.

27. The method of claim 26, wherein the image is a video, still image, real-time video, or any combination thereof.

28. The method of claim 27, wherein the subject or the image is visualized or displayed during a surgical procedure conducted on the subject due to the imaging agent.

29. The method of claim 28, wherein the imaging agent is administered during or before surgery for endometriosis diagnosis, staging, and / or excision.

30. The method of claim 24, wherein the formulation is administered intravaginally or intravenously.

31. The method of claim 24, wherein the imaging in (b) comprises in vivo fluorescence imaging, Magnetic Resonance Imaging (MRI), Positron Emission Tomography (PET), Single Photon Emission Computed Tomography (SPECT), ultrasound imaging, Computed Tomography (CT), or photoacoustic imaging.

32. The method of claim 24, wherein the subject is human, a cultured cell line, or an ex vivo tissue extracted from an animal.

33. The method of claim 24, wherein the imaging comprises whole body imaging.

34. The method of claim 24, wherein the second [3-catenin-binding peptide or peptidomimetic comprises an amino acid sequence according to Formula I:R-X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12- X13-X14-X15-X16 (Formula I), wherein:R is NH2, acetylation, stearic acid, palmitic acid, myristic acid, lauric acid, a Ci-Cs hydrocarbon, a Ci-Cs fatty acid, or null;Xi is M, G, beta alanine, norleucine, norvaline, or null;X2is W, N-methyl W, R, Y, F, citrulline, or K,X3is 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;X5is S, alpha methyl S, K, D, Orn, T, or E;X6is I, Chg, H, or L;X7is L or I;X8is D, N, E, or Q;Xg is D, E, K, Q, or Orn;X is H or methyl-H;X11 is V, alpha methyl V, Chg, L, I, or norvaline;X12 is Q, Aib, S, R, or N;X13 is R, K, citrulline, Orn, D, or E;X14 is V, I, L, or norvaline;X15 is W, Y, or F; andX16 is R, G, or null.

35. The method of claim 34, wherein the second [3-catenin-binding peptide or peptidomimetic has the amino acid sequence Oct-W-NMeA-Q-Orn*-ILDE*H-Chg-QRVWG-NH2 (SEQ ID NO:491).

36. The method of claim 24, wherein the second [3-catenin-binding peptide or peptidomimetic comprises an amino acid sequence according to Formula II:X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12- X13-X14-X15-X16 (Formula II), wherein:Xi is M or null;X2is S, I, G, T, A, L, or null;X3is R, K or null;X4is a positively-charged amino acid, citrulline, Orn, D, E, 8-aminooctanoic acid, or an amino carboxylic acid with between 4 and 12 carbons;X5is M, Norleucine, Orn, D, E, K, H, R, K, 8-aminooctanoic acid, an amino carboxylic acid with between 4 and 12 carbons or null;Xe is W, Y, F, or N-methyl A;X? is F, I, L, Chg, Cha, or Tie;Xs is L, I, or A;X9 is L, I, or A;Xw is C, S, A, Abu, C(me), or S(Bzl);X11 is F, H, A, K, E, Chg, Cng, or Orn;X12 is W, Y, A, or F;X13 is G, GABA, or null;X14 is V, I, L, norvaline or null;X15 is W, Y, F, or null; andX16 is R, G, or null.

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