Methods of reducing adhesions and inflammation using corticotropin releasing hormone receptor antagonists

Abstract

The present disclosure provides a method of reducing formation of one or more adhesions in a patient by administering a CRH1 receptor antagonist to the patient. Additional methods of administering a CRH1 receptor antagonist to patients are also provided, including methods of reducing gastrointestinal damage, reducing inflammation, and reducing pain.

Description

71900-429670METHODS OF REDUCING ADHESIONS AND INFLAMMATION USING CORTICOTROPIN RELEASING HORMONE RECEPTOR ANTAGONISTSCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit under 35 U.S.C. § 119(e) of U.S. Provisional Application Serial No. 63 / 688,026, filed on August 28, 2024, the entire disclosures of which is incorporated herein by reference.BACKGROUND AND SUMMARY

[0002] Formation of adhesions in patients is a complicated and costly medical issue. For instance, the post-operative adhesions that form after abdominal surgery, whether planned or emergency, contribute to the risk of bowel obstruction, chronic pain, and infertility7in patients. The cost of treating post-surgical adhesions surpasses $1 billion in the United States, according to the National Institutes of Health.

[0003] Unfortunately, performing recurrent operations increases the incidence of adhesions. For instance, the chronic gy necological disease endometriosis, a ty pical cycle of care often involves repeated surgical operations to ablate or excise the ectopic tissues, thus significantly increasing the likelihood of adhesion formation. The rate of re-operation in women with endometriosis is exceptionally high, with approximately 62% of women undergoing two or more interventions in less than five years of treatment. Therefore, women with endometriosis can be exposed to a never-ending cycle of adhesion formation and surgical intervention, followed by adhesion re-formation.

[0004] Additionally, patients in which adhesions are common are also increasingly- recognized to exhibit undesirable gastrointestinal symptoms, including abdominal pain, bloating, constipation, diarrhea, and rectal bleeding. Moreover, inflammatory7changes can be observed in this patient population.

[0005] Although adhesions can be treated by surgical removal, there are no current options for pharmaceutical treatments, including oral pharmacological treatments. Physical barrier products have been developed to cover tissues during surgery7in an attempt to minimize adhesions but are of limited efficacy . Therefore, there exists a need for new therapeutics for the prevention and treatment of adhesions as well as the accompanying inflammatory and gastrointestinal damage in patients.

[0006] Accordingly, the present disclosure provides methods for reducing formation of adhesions in a patient by administering a CRH1 receptor antagonist to the patient. In addition, methods for reducing gastrointestinal damage and reducing inflammation in a patient by71900-429670 administering a CRH1 receptor antagonist to the patient are also provided. The methods provide benefits to a diverse patient population and can be advantageously be utilized as prophylactic treatments against adhesions in patients.

[0007] Other objects, features and advantages of the present disclosure will become apparent from the following detailed description. It should be understood, however, that the detailed description and the specific examples, while indicating specific embodiments of the invention, are given by way of illustration only, since various changes and modifications within the spirit and scope of the invention will become apparent to those skilled in the art from this detailed description.BRIEF DESCRIPTIONS OF THE DRAWINGS

[0008] The detailed description particularly refers to the accompanying figures in which:

[0009] FIGURE 1 illustrates the experimental timelines for the results of Examples 2-5.

[0010] FIGURES 2A-2I show developed endometriosis vesicle characteristics. As shown in FIGS. 2A-2I, when pexacerfont is administered following surgery, there are no changes in the size and number of developed endometriosis vesicles, regardless of the dose (TAP and ENT).

[0011] FIGURES 3A-3C depict the developed endometriosis vesicle characteristics. Pexacerfont, given after endometriosis develops (25 days after surgery), produces no change in weight or area of developed vesicles (STT).

[0012] FIGURES 4A-4C depict macroscopic scores, with FIG. 4A displaying the TAP project and both FIG. 4B and FIG. 4C displaying the ENT project. A dose of 10 mg / kg of pexacerfont administered after surgery show ed a decrease in macroscopic score, mostly observed as a decrease of adhesion severity. Table 1 shows the individual values for the four parameters that constitute the macroscopic score.

[0013] FIGURE 5 shows the macroscopic scores for the STT project. Pexacerfont given 25 days after surgery' in both acute and chronic settings prevented the increase in the macroscopic score when assessed at 60 days after surgery. Table 2 shows the individual values for the four parameters that constitute each macroscopic score.

[0014] FIGURE 6 presents the percentage of rats per group that developed any level of adhesions (i.e., a score of 1 or 2).

[0015] FIGURES 7A-7B show' reduction in pain scores in animals. FIG. 7A shows the inflammatory pain reaction at day 60 after endometriosis onset for the STT project. FIG. 7B show s the correlation of the pain scores with the total macroscopic score for each rat.71900-429670

[0016] FIGURE 8 illustrates the experimental timelines for Examples 7-13 in which antagonizing CRHR-1 provides a reduction in adhesions.

[0017] FIGURES 9A-9D show the endometriosis development measures for Examples 7-13. antagonizing CRHR-1 provides a reduction in adhesions.

[0018] FIGURES 10A-10E depict nerve grow th factor (NGF), Ki67, vascular endothelial growth factor (VEGF) as measured in Examples 7-13. As shown in FIGS. 10A- 10E, antagonizing CRHR-1 provides a reduction in adhesions.

[0019] FIGURES 11A-11C present nociceptive tests of Examples 7-13. As shown in FIGS. 11A-11C, antagonizing the CRHR-1 provides a reduction in pain.

[0020] FIGURES 12A-12C exhibit LH, FSH, GNRH as monitored in Examples 7-13.

[0021] FIGURES 13A-13C quantifies the cytokines as measured by PCR in the vesicles.

[0022] FIGURES 14A-14B depict the milliplex rat pituitary panel of cytokines to measure IL-6 and IL-1 (3.

[0023] FIGURES 15A-15D exhibit the correlation of leptin compared to animal weight as measured for Examples 7-13.

[0024] FIGURE 16 illustrates histopathology of vesicles as measured in Examples 7-13.

[0025] FIGURE 17 illustrates the experimental timeline for Example 14, displaying the preventive treatment against post-surgical abdominal adhesions.

[0026] FIGURE 18 depicts the percentage w eight change of the rats as measured inExample 14.

[0027] FIGURES 19A-19B show the Von Frey test for mechanical allodynia (FIG. 19A) and the hot plate test for thermal hyperalgesia (FIG. 19B) in the abrasion rat model described in Example 14. Individual data points are displayed, with full symbols representing the male rats and clear symbols representing the female rats.

[0028] FIGURES 20A-20B exhibit the overall adhesion score (FIG. 20A) and adhesion score only in the colon (FIG. 20B).

[0029] FIGURE 21 depicts analysis of pexacerfont administration on cecum damage in animals.

[0030] FIGURE 22 presents the macroscopic score for colonic damage in animals. The individual values are exhibited in Table 3.

[0031] FIGURES 23A-23B show microscopic scores for colon in animals (FIG. 23 A) and microscopic scores for cecum in animals (FIG. 23B).71900-429670

[0032] FIGURES 24A-24D present fibrotic scoring in the colon of animals (FIG. 24A) and fibrotic scoring in the cecum of animals (FIG. 24B). Further, FIG. 24C shows the final grading score in colon and FIG. 24D shows the final grading score for the cecum.

[0033] FIGURES 25A-25B show Mast cell counts observed in the distal colon of animals (FIG. 25 A) and in the cecum of animals (FIG. 25B).DETAILED DESCRIPTION

[0034] Various embodiments of the invention are described herein as follows. In an illustrative aspect, a method of reducing formation of one or more adhesions in a patient is provided. The method comprises the step of administering a CRH1 receptor antagonist to the patient, wherein the administration of the CRH1 receptor antagonist reduces formation of adhesions.

[0035] In an embodiment, the patient is male. In an embodiment, the patient is female.

[0036] In an embodiment, the one or more adhesions comprises an endometriosis- related adhesion. In an embodiment, the one or more adhesions comprises a pelvic adhesion. In an embodiment, the one or more adhesions comprises a gastrointestinal adhesion. In an embodiment, the one or more adhesions comprises a post-surgical adhesion.

[0037] In an embodiment, the one or more adhesions are induced by a surgical intervention on the patient. In an embodiment, the surgical intervention is performed in the peritoneal cavity of the patient.

[0038] In an embodiment, administering the CRH1 receptor antagonist maintains FSH / LH ratio in the patient.

[0039] In an embodiment, administering the CRH1 receptor antagonist is an acute administration to the patient. In an embodiment, administering the CRH1 receptor antagonist is a chronic administration to the patient. In an embodiment, administering the CRH1 receptor antagonist is a prophylactic administration to the patient.

[0040] In an embodiment, administering the CRH1 receptor antagonist is an oral administration to the patient. In an embodiment, administering the CRH1 receptor antagonist is a parenteral administration to the patient.

[0041] In an embodiment, the CRH1 receptor antagonist is pexacerfont. In an embodiment, the CRH1 receptor antagonist is antalarmin. In an embodiment, the CRH1 receptor antagonist is emicerfont. In an embodiment, the CRH1 receptor antagonist is crinecerfont. In an embodiment, the CRH1 receptor antagonist is tildacerfont. In an embodiment, the CRH1 receptor antagonist is verucerfont (GSK-561679). In an embodiment, the CRH1 receptor antagonist is LWH-234. In an embodiment, the CRH1 receptor antagonist71900-429670 is CP-154,526. In an embodiment, the CRH1 receptor antagonist is NBI-27914. In an embodiment, the CRH1 receptor antagonist is R-121,919. In an embodiment, the CRH1 receptor antagonist is GW-876008. In an embodiment, the CRH1 receptor antagonist is CRA- 1000. In an embodiment, the CRH1 receptor antagonist is NB1-35965. In an embodiment, the CRH1 receptor antagonist is R121919 (NBI-30775). In an embodiment, the CRH1 receptor antagonist is CP-316,311. In an embodiment, the CRH1 receptor antagonist is R317573 (JNJ- 19567470; CRA-5626). In an embodiment, the CRH1 receptor antagonist is ONO-2333Ms. In an embodiment, the CRH1 receptor antagonist is GSK586529. In an embodiment, the CRH1 receptor antagonist is a-helical CRH (9-41). In an embodiment, the CRH1 receptor antagonist is astressin-2B.

[0042] In an illustrative aspect, a method of reducing gastrointestinal damage in a patient is provided. The method comprises the step of administering a CRH1 receptor antagonist to the patient, wherein the administration of the CRH1 receptor antagonist reduces gastrointestinal damage. Means to evaluate the reduction of gastrointestinal damage in a patient are described herein and / or known to the skilled artisan.

[0043] In an embodiment, the patient is male. In an embodiment, the patient is female.

[0044] In an embodiment, the gastrointestinal damage comprises gastrointestinal adhesions. In an embodiment, the gastrointestinal damage comprises endometriosis-induced gastrointestinal damage.

[0045] In an embodiment, the gastrointestinal damage comprises fibrosis. In an embodiment, the fibrosis comprises colonic fibrosis in the patient. In an embodiment, the fibrosis comprises cecal fibrosis in the patient.

[0046] In an embodiment, administering the CRH1 receptor antagonist maintains follicle-stimulating hormone (FSH)Zlutenizing hormone (LH) ratio in the patient.

[0047] In an embodiment, administering the CRH1 receptor antagonist is an acute administration to the patient. In an embodiment, administenng the CRH1 receptor antagonist is a chronic administration to the patient. In an embodiment, administering the CRH1 receptor antagonist is a prophylactic administration to the patient.

[0048] In an embodiment, administering the CRH1 receptor antagonist is an oral administration to the patient. In an embodiment, administering the CRH1 receptor antagonist is a parenteral administration to the patient.

[0049] In an embodiment, the CRH1 receptor antagonist is pexacerfont. In an embodiment, the CRH1 receptor antagonist is antalarmin. In an embodiment, the CRH1 receptor antagonist is emicerfont. In an embodiment, the CRH1 receptor antagonist is crinecerfont. In an embodiment, the CRH1 receptor antagonist is tildacerfont. In an71900-429670 embodiment, the CRH1 receptor antagonist is verucerfont (GSK-561679). In an embodiment, the CRH1 receptor antagonist is LWH-234. In an embodiment, the CRH1 receptor antagonist is CP-154,526. In an embodiment, the CRH1 receptor antagonist is NBI-27914. In an embodiment, the CRH1 receptor antagonist is R-121.919. In an embodiment, the CRH1 receptor antagonist is GW-876008. In an embodiment, the CRH1 receptor antagonist is CRA- 1000. In an embodiment, the CRH1 receptor antagonist is NBI-35965. In an embodiment, the CRH1 receptor antagonist is R121919 (NBI-30775). In an embodiment, the CRH1 receptor antagonist is CP-316,311. In an embodiment, the CRH1 receptor antagonist is R317573 (JNJ- 19567470; CRA-5626). In an embodiment, the CRH1 receptor antagonist is ONO-2333Ms. In an embodiment, the CRH1 receptor antagonist is GSK586529. In an embodiment, the CRH1 receptor antagonist is a-helical CRH (9-41). In an embodiment, the CRH1 receptor antagonist is astressin-2B.

[0050] In an illustrative aspect, a method of reducing inflammation in a patient is provided. The method comprises the step of administering a CRH1 receptor antagonist to the patient, wherein the administration of the CRH1 receptor antagonist reduces one or more inflammatory' signals in the patient. Means to evaluate the reduction of inflammation in a patient are described herein and / or known to the skilled artisan.

[0051] In an embodiment, the patient is male. In an embodiment, the patient is female.

[0052] In an embodiment, the one or more inflammatory signals comprises a decrease in an inflammatory cytokine. In an embodiment, the inflammatory cytokine is IL-6. In an embodiment, the inflammatory cytokine is IL-ip. In an embodiment, the inflammatory cytokine is TNF-a. In an embodiment, the one or more inflammatory signals comprises a decrease in mast cells.

[0053] In an embodiment, the inflammation comprises colonic inflammation. In an embodiment, the inflammation comprises cecal inflammation.

[0054] In an embodiment, the inflammation comprises fibrosis. In an embodiment, the fibrosis comprises colonic fibrosis in the patient. In an embodiment, the fibrosis comprises cecal fibrosis in the patient.

[0055] In an embodiment, administering the CRH1 receptor antagonist maintains FSH / LH ratio in the patient.

[0056] In an embodiment, administering the CRH1 receptor antagonist is an acute administration to the patient. In an embodiment, administering the CRH1 receptor antagonist is a chronic administration to the patient. In an embodiment, administering the CRH1 receptor antagonist is a prophylactic administration to the patient.71900-429670

[0057] In an embodiment, administering the CRH1 receptor antagonist is an oral administration to the patient. In an embodiment, administering the CRH1 receptor antagonist is a parenteral administration to the patient.

[0058] In an embodiment, the CRH1 receptor antagonist is pexacerfont. In an embodiment, the CRH1 receptor antagonist is antalarmin. In an embodiment, the CRH1 receptor antagonist is emicerfont. In an embodiment, the CRH1 receptor antagonist is crinecerfont. In an embodiment, the CRH1 receptor antagonist is tildacerfont. In an embodiment, the CRH1 receptor antagonist is verucerfont (GSK-561679). In an embodiment, the CRH1 receptor antagonist is LWH-234. In an embodiment, the CRH1 receptor antagonist is CP-154,526. In an embodiment, the CRH1 receptor antagonist is NBI-27914. In an embodiment, the CRH1 receptor antagonist is R-121,919. In an embodiment, the CRH1 receptor antagonist is GW-876008. In an embodiment, the CRH1 receptor antagonist is CRA- 1000. In an embodiment, the CRH1 receptor antagonist is NBI-35965. In an embodiment, the CRH1 receptor antagonist is R121919 (NBI-30775). In an embodiment, the CRH1 receptor antagonist is CP-316,311. In an embodiment, the CRH1 receptor antagonist is R317573 (JNJ- 19567470; CRA-5626). In an embodiment, the CRH1 receptor antagonist is ONO-2333Ms. In an embodiment, the CRH1 receptor antagonist is GSK586529. In an embodiment, the CRH1 receptor antagonist is a-helical CRH (9-41). In an embodiment, the CRH1 receptor antagonist is astressin-2B.

[0059] In an illustrative aspect, a method of reducing pain in a patient is provided. The method comprises the step of administering a CRH1 receptor antagonist to the patient, wherein the administration of the CRH1 receptor antagonist reduces one or more pain symptoms in the patient. Means to evaluate the reduction of pain in a patient are described herein and / or known to the skilled artisan.

[0060] In an embodiment, the patient is male. In an embodiment, the patient is female.

[0061] In an embodiment, the reduction in one or more pain symptoms comprises a reduction in inflammatory pain. In an embodiment, the reduction in inflammatory pain a decrease in an inflammatory' cytokine. In an embodiment, the inflammatory cytokine is IL-6. In an embodiment, the inflammatory' cytokine is IL-ip. In an embodiment, the inflammatory' cytokine is TNF-a. In an embodiment, the reduction in one or more pain symptoms comprises a reduction in mechanical pain.

[0062] In an embodiment, administering the CRH1 receptor antagonist maintains FSH / LH ratio in the patient. In an embodiment, administering the CRH1 receptor antagonist is an acute administration to the patient. In an embodiment, administering the CRH1 receptor antagonist is a chronic administration to the patient. In an embodiment, administering the71900-429670CRH1 receptor antagonist is a prophylactic administration to the patient. In an embodiment, administering the CRH1 receptor antagonist is an oral administration to the patient. In an embodiment, administering the CRH1 receptor antagonist is a parenteral administration to the patient.

[0063] In an embodiment, the CRH1 receptor antagonist is pexacerfont. In an embodiment, the CRH1 receptor antagonist is antalarmin. In an embodiment, the CRH1 receptor antagonist is emicerfont. In an embodiment, the CRH1 receptor antagonist is crinecerfont. In an embodiment, the CRH1 receptor antagonist is tildacerfont. In an embodiment, the CRH1 receptor antagonist is verucerfont (GSK-561679). In an embodiment, the CRH1 receptor antagonist is LWH-234. In an embodiment, the CRH1 receptor antagonist is CP-154,526. In an embodiment, the CRH1 receptor antagonist is NBI-27914. In an embodiment, the CRH1 receptor antagonist is R-121,919. In an embodiment, the CRH1 receptor antagonist is GW-876008. In an embodiment, the CRH1 receptor antagonist is CRA- 1000. In an embodiment, the CRH1 receptor antagonist is NBI-35965. In an embodiment, the CRH1 receptor antagonist is R121919 (NBI-30775). In an embodiment, the CRH1 receptor antagonist is CP-316,311. In an embodiment, the CRH1 receptor antagonist is R317573 (JNJ- 19567470; CRA-5626). In an embodiment, the CRH1 receptor antagonist is ONO-2333Ms. In an embodiment, the CRH1 receptor antagonist is GSK586529. In an embodiment, the CRH1 receptor antagonist is a-helical CRH (9-41). In an embodiment, the CRH1 receptor antagonist is astressin-2B.

[0064] The following numbered embodiments are contemplated and are non-limiting:1. A method of reducing formation of one or more adhesions in a patient, the method comprising the step of administering a CRH1 receptor antagonist to the patient, wherein the administration of the CRH1 receptor antagonist reduces formation of adhesions.2. The method of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the patient is male.3. The method of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the patient is female.4. The method of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the one or more adhesions comprises an endometriosis-related adhesion.5. The method of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the one or more adhesions comprises a pelvic adhesion.6. The method of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the one or more adhesions comprises a gastrointestinal adhesion.71900-4296707. The method of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the one or more adhesions comprises a post-surgical adhesion.8. The method of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the one or more adhesions are induced by a surgical intervention on the patient.9. The method of clause 8, any other suitable clause, or any combination of suitable clauses, wherein the surgical intervention is performed in the peritoneal cavity of the patient.10. The method of clause 1, any other suitable clause, or any combination of suitable clauses, wherein administering the CRH1 receptor antagonist maintains FSH / LH ratio in the patient.11. The method of clause 1, any other suitable clause, or any combination of suitable clauses, wherein administering the CRH1 receptor antagonist is an acute administration to the patient.12. The method of clause 1, any other suitable clause, or any combination of suitable clauses, wherein administering the CRH1 receptor antagonist is a chronic administration to the patient.13. The method of clause 1, any other suitable clause, or any combination of suitable clauses, wherein administering the CRH1 receptor antagonist is a prophylactic administration to the patient.14. The method of clause 1, any other suitable clause, or any combination of suitable clauses, wherein administering the CRH1 receptor antagonist is an oral administration to the patient.15. The method of clause 1, any other suitable clause, or any combination of suitable clauses, wherein administering the CRH1 receptor antagonist is a parenteral administration to the patient.16. The method of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the CRH1 receptor antagonist is pexacerfont.17. The method of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the CRH1 receptor antagonist is antalarmin.18. The method of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the CRH1 receptor antagonist is emicerfont.19. The method of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the CRH1 receptor antagonist is crinecerfont.20. The method of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the CRH1 receptor antagonist is tildacerfont.71900-42967021. The method of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the CRH1 receptor antagonist is verucerfont (GSK-561679).22. The method of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the CRH1 receptor antagonist is LWH-234.23. The method of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the CRH1 receptor antagonist is CP-154,526.24. The method of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the CRH1 receptor antagonist is NBI-27914.25. The method of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the CRH1 receptor antagonist is R-121,919.26. The method of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the CRH1 receptor antagonist is GW-876008.27. The method of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the CRH1 receptor antagonist is CRA-1000.28. The method of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the CRH1 receptor antagonist is NBI-35965, or R121919 (NBI-30775), or CP- 316,311. or R317573 (JNJ- 19567470; CRA-5626), or ONO-2333Ms, or GSK586529, or a- helical CRH (9-41), or astressin-2B.29. A method of reducing gastrointestinal damage in a patient, the method comprising the step of administering a CRH1 receptor antagonist to the patient, wherein the administration of the CRH1 receptor antagonist reduces gastrointestinal damage.30. The method of clause 29, any other suitable clause, or any combination of suitable clauses, wherein the patient is male.31. The method of clause 29, any other suitable clause, or any combination of suitable clauses, wherein the patient is female.32. The method of clause 29, any other suitable clause, or any combination of suitable clauses, wherein the gastrointestinal damage comprises gastrointestinal adhesions.33. The method of clause 29, any other suitable clause, or any combination of suitable clauses, wherein the gastrointestinal damage comprises endometriosis-induced gastrointestinal damage.34. The method of clause 29, any other suitable clause, or any combination of suitable clauses, wherein the gastrointestinal damage comprises fibrosis.35. The method of clause 34, any other suitable clause, or any combination of suitable clauses, wherein the fibrosis comprises colonic fibrosis in the patient.71900-42967036. The method of clause 34, any other suitable clause, or any combination of suitable clauses, wherein the fibrosis comprises cecal fibrosis in the patient.37. The method of clause 29, any other suitable clause, or any combination of suitable clauses, wherein administering the CRH1 receptor antagonist maintains FSH / LH ratio in the patient.38. The method of clause 29, any other suitable clause, or any combination of suitable clauses, wherein administering the CRH1 receptor antagonist is an acute administration to the patient.39. The method of clause 29, any other suitable clause, or any combination of suitable clauses, wherein administering the CRH1 receptor antagonist is a chronic administration to the patient.40. The method of clause 29, any other suitable clause, or any combination of suitable clauses, wherein administering the CRH1 receptor antagonist is a prophylactic administration to the patient.41. The method of clause 29, any other suitable clause, or any combination of suitable clauses, wherein administering the CRH1 receptor antagonist is an oral administration to the patient.42. The method of clause 29, any other suitable clause, or any combination of suitable clauses, wherein administering the CRH1 receptor antagonist is a parenteral administration to the patient.43. The method of clause 29, any other suitable clause, or any combination of suitable clauses, wherein the CRH1 receptor antagonist is pexacerfont.44. The method of clause 29, any other suitable clause, or any combination of suitable clauses, wherein the CRH1 receptor antagonist is antalarmin.45. The method of clause 29, any other suitable clause, or any combination of suitable clauses, wherein the CRH1 receptor antagonist is emicerfont.46. The method of clause 29, any other suitable clause, or any combination of suitable clauses, wherein the CRH1 receptor antagonist is crinecerfont.47. The method of clause 29, any other suitable clause, or any combination of suitable clauses, wherein the CRH1 receptor antagonist is tildacerfont.48. The method of clause 29, any other suitable clause, or any combination of suitable clauses, wherein the CRH1 receptor antagonist is verucerfont (GSK-561679).49. The method of clause 29, any other suitable clause, or any combination of suitable clauses, wherein the CRH1 receptor antagonist is LWH-234.71900-42967050. The method of clause 29, any other suitable clause, or any combination of suitable clauses, wherein the CRH1 receptor antagonist is CP-154,526.51. The method of clause 29, any other suitable clause, or any combination of suitable clauses, wherein the CRH1 receptor antagonist is NBI-27914.52. The method of clause 29, any other suitable clause, or any combination of suitable clauses, wherein the CRH1 receptor antagonist is R-121,919.53. The method of clause 29, any other suitable clause, or any combination of suitable clauses, wherein the CRH1 receptor antagonist is GW-876008.54. The method of clause 29, any other suitable clause, or any combination of suitable clauses, wherein the CRH1 receptor antagonist is CRA-1000.55. The method of clause 29, any other suitable clause, or any combination of suitable clauses, wherein the CRH1 receptor antagonist is NBI-35965, or R121919 (NBI-30775), or CP- 316,311, or R317573 (JNJ- 19567470; CRA-5626), or ONO-2333Ms, or GSK586529, or a- helical CRH (9-41), or astressin-2B.56. A method of reducing inflammation in a patient, the method comprising the step of administering a CRH1 receptor antagonist to the patient, wherein the administration of the CRH1 receptor antagonist reduces one or more inflammatory signals in the patient.57. The method of clause 56, any other suitable clause, or any combination of suitable clauses, wherein the patient is male.58. The method of clause 56, any other suitable clause, or any combination of suitable clauses, wherein the patient is female.59. The method of clause 56, any other suitable clause, or any combination of suitable clauses, wherein the one or more inflammatory signals comprises a decrease in an inflammatory cytokine.60. The method of clause 59, any other suitable clause, or any combination of suitable clauses, wherein the inflammatory cytokine is IL-6.61. The method of clause 59, any other suitable clause, or any combination of suitable clauses, wherein the inflammatory cytokine is IL-10.62. The method of clause 59, any other suitable clause, or any combination of suitable clauses, wherein the inflammatory cytokine is TNF-a.63. The method of clause 56, any other suitable clause, or any combination of suitable clauses, wherein the one or more inflammatory signals comprises a decrease in mast cells.64. The method of clause 56, any other suitable clause, or any combination of suitable clauses, wherein the inflammation comprises colonic inflammation.71900-42967065. The method of clause 56, any other suitable clause, or any combination of suitable clauses, wherein the inflammation comprises cecal inflammation.66. The method of clause 56, any other suitable clause, or any combination of suitable clauses, wherein the inflammation comprises fibrosis.67. The method of clause 66, any other suitable clause, or any combination of suitable clauses, wherein the fibrosis comprises colonic fibrosis in the patient.68. The method of clause 66, any other suitable clause, or any combination of suitable clauses, wherein the fibrosis comprises cecal fibrosis in the patient.69. The method of clause 56, any other suitable clause, or any combination of suitable clauses, wherein administering the CRH1 receptor antagonist maintains FSH / LH ratio in the patient.70. The method of clause 56, any other suitable clause, or any combination of suitable clauses, wherein administering the CRH1 receptor antagonist is an acute administration to the patient.71. The method of clause 56, any other suitable clause, or any combination of suitable clauses, wherein administering the CRH1 receptor antagonist is a chronic administration to the patient.72. The method of clause 56, any other suitable clause, or any combination of suitable clauses, wherein administering the CRH1 receptor antagonist is a prophylactic administration to the patient.73. The method of clause 56, any other suitable clause, or any combination of suitable clauses, wherein administering the CRH1 receptor antagonist is an oral administration to the patient.74. The method of clause 56, any other suitable clause, or any combination of suitable clauses, wherein administering the CRH1 receptor antagonist is a parenteral administration to the patient.75. The method of clause 56, any other suitable clause, or any combination of suitable clauses, wherein the CRH1 receptor antagonist is pexacerfont.76. The method of clause 56, any other suitable clause, or any combination of suitable clauses, wherein the CRH1 receptor antagonist is antalarmin.77. The method of clause 56, any other suitable clause, or any combination of suitable clauses, wherein the CRH1 receptor antagonist is emicerfont.78. The method of clause 56, any other suitable clause, or any combination of suitable clauses, wherein the CRH1 receptor antagonist is crinecerfbnt.71900-42967079. The method of clause 56, any other suitable clause, or any combination of suitable clauses, wherein the CRH1 receptor antagonist is tildacerfont.80. The method of clause 56, any other suitable clause, or any combination of suitable clauses, wherein the CRH1 receptor antagonist is verucerfont (GSK-561679).81. The method of clause 56, any other suitable clause, or any combination of suitable clauses, wherein the CRH1 receptor antagonist is LWH-234.82. The method of clause 56, any other suitable clause, or any combination of suitable clauses, wherein the CRH1 receptor antagonist is CP- 154,526.83. The method of clause 56, any other suitable clause, or any combination of suitable clauses, wherein the CRH1 receptor antagonist is NBI-27914.84. The method of clause 56, any other suitable clause, or any combination of suitable clauses, wherein the CRH1 receptor antagonist is R-121,919.85. The method of clause 56, any other suitable clause, or any combination of suitable clauses, wherein the CRH1 receptor antagonist is GW-876008.86. The method of clause 56, any other suitable clause, or any combination of suitable clauses, wherein the CRH1 receptor antagonist is CRA-1000.87. The method of clause 56, any other suitable clause, or any combination of suitable clauses, wherein the CRH1 receptor antagonist is NBI-35965, or R121919 (NBI-30775), or CP- 316,311, or R317573 (JNJ- 19567470; CRA-5626), or ONO-2333Ms, or GSK586529, or a- helical CRH (9-41), or astressin-2B.88. A method of reducing pain in a patient, the method comprising the step of administering a CRH1 receptor antagonist to the patient, wherein the administration of the CRH1 receptor antagonist reduces one or more pain symptoms in the patient.89. The method of clause 88, any other suitable clause, or any combination of suitable clauses, wherein the patient is male.90. The method of clause 88, any other suitable clause, or any combination of suitable clauses, wherein the patient is female.91. The method of clause 88, any other suitable clause, or any combination of suitable clauses, wherein the reduction in one or more pain symptoms comprises a reduction in inflammatory pain.92. The method of clause 91, any other suitable clause, or any combination of suitable clauses, wherein the reduction in inflammatory pain a decrease in an inflammatory cytokine.93. The method of clause 91, any other suitable clause, or any combination of suitable clauses, wherein the inflammatory cytokine is IL-6.71900-42967094. The method of clause 91, any other suitable clause, or any combination of suitable clauses, wherein the inflammatory cytokine is IL-10.95. The method of clause 91, any other suitable clause, or any combination of suitable clauses, wherein the inflammatory cytokine is TNF-a.96. The method of clause 88, any other suitable clause, or any combination of suitable clauses, wherein the reduction in one or more pain symptoms comprises a reduction in mechanical pain.97. The method of clause 88, any other suitable clause, or any combination of suitable clauses, wherein administering the CRH1 receptor antagonist maintains FSH / LH ratio in the patient.98. The method of clause 88, any other suitable clause, or any combination of suitable clauses, wherein administering the CRH1 receptor antagonist is an acute administration to the patient.99. The method of clause 88, any other suitable clause, or any combination of suitable clauses, wherein administering the CRH1 receptor antagonist is a chronic administration to the patient.100. The method of clause 88, any other suitable clause, or any combination of suitable clauses, wherein administering the CRH1 receptor antagonist is a prophylactic administration to the patient.101. The method of clause 88, any other suitable clause, or any combination of suitable clauses, wherein administering the CRH1 receptor antagonist is an oral administration to the patient.102. The method of clause 88, any other suitable clause, or any combination of suitable clauses, wherein administering the CRH1 receptor antagonist is a parenteral administration to the patient.103. The method of clause 88, any other suitable clause, or any combination of suitable clauses, wherein the CRH1 receptor antagonist is pexacerfont.104. The method of clause 88, any other suitable clause, or any combination of suitable clauses, wherein the CRH1 receptor antagonist is antalarmin.105. The method of clause 88, any other suitable clause, or any combination of suitable clauses, wherein the CRH1 receptor antagonist is emicerfont.106. The method of clause 88, any other suitable clause, or any combination of suitable clauses, wherein the CRH1 receptor antagonist is crinecerfont.107. The method of clause 88, any other suitable clause, or any combination of suitable clauses, wherein the CRH1 receptor antagonist is tildacerfont.71900-429670108. The method of clause 88, any other suitable clause, or any combination of suitable clauses, wherein the CRH1 receptor antagonist is verucerfont (GSK-561679).109. The method of clause 88, any other suitable clause, or any combination of suitable clauses, wherein the CRH1 receptor antagonist is LWH-234.1 10. The method of clause 88, any other suitable clause, or any combination of suitable clauses, wherein the CRH1 receptor antagonist is CP-154,526.111. The method of clause 88, any other suitable clause, or any combination of suitable clauses, wherein the CRH1 receptor antagonist is NBI-27914.1 12. The method of clause 88, any other suitable clause, or any combination of suitable clauses, wherein the CRH1 receptor antagonist is R-121,919.113. The method of clause 88, any other suitable clause, or any combination of suitable clauses, wherein the CRH1 receptor antagonist is GW-876008.1 14. The method of clause 88, any other suitable clause, or any combination of suitable clauses, wherein the CRH1 receptor antagonist is CRA-1000.115. The method of clause 88, any other suitable clause, or any combination of suitable clauses, wherein the CRH1 receptor antagonist is NBI-35965, or R121919 (NBI- 30775), or CP-316,311, or R317573 (JNJ- 19567470; CRA-5626), or ONO-2333Ms. or GSK586529, or a-helical CRH (9-41), or astressin-2B.EXAMPLESEXAMPLE 1Materials and Methods for Examples 2-5Animals and groups

[0065] For Examples 2-6, 60-day-old female Sprague Dawley rats weighing 190-230 grams were used. The rats were purchased from Ponce Research Institute Animal Facilities and housed two per cage with a 12-hour light / dark cycle and access to food and water. The rats were monitored for weight and estrous cycles daily during the therapeutic administration period and weighed twice weekly afterward.Experimental approach

[0066] The onset of endometriosis-associated hyperalgesia occurs around day 28 after surgery’. Therefore, 28 days was the time point selected for measuring endometriosis development, adhesions, diarrhea, and colonic damage for the TAP and ENT projects. The 7-71900-429670 day time point in the ENT project was included to evaluate the short-term effects of the therapeutic (which may correspond to an acute phase).

[0067] For the STT project, endometriosis was allowed to progress until Day 60. In addition, a more prolonged timeframe provided evaluation of repeated therapeutic administration effects (e g., chronic groups). FIG. 1 illustrates the experimental timelines and groups used in Examples 2-5.Surgical induction of endometriosis

[0068] Endometriosis was induced surgically in animals following established protocols. Briefly, the distal 2 cm of the right uterine hom was removed and opened, cutting four pieces of the uterine hom of 2 mm x 2 mm. These implants were sutured with silk sutures next to the mesenteric vessels of the large intestine, with the serosal side facing toward the mesentery and the endometrial side facing the peritoneum. Rats in the sham surgery group received sutures without uterine tissue.Therapeutic dose and administration

[0069] Animals received a 7-day consecutive oral doses of pexacerfont suspended in 0.5% methylcellulose for the TAP and ENT projects (Examples 2-5). Pexacerfont was administered at a concentration of 3 or 10 mg / kg (1 mg / rnL volume) and injected inside an oyster cracker. Rats were previously trained to eat the oyster cracker as this method reduces the animals’ stress compared to administering the therapeutic by oral gavage.

[0070] Pexacerfont or vehicle dosing was administered on the morning following surgery and administration continued for seven consecutive days. During this time, the estrous cycle of the animals was monitored via vaginal smears, and the animals were weighed daily. For the STT project, either pexacerfont or vehicle was administered to animals starting on day 25 after surgical induction of endometriosis (see FIG. 1). A positive control group receiving 14 mg / kg of elagolix (dissolved in PBS and methylcellulose 0.5%) was also included.Endometriosis development quantification

[0071] On either day 7 or day 28 after surgical induction of endometriosis (for TAP and ENT projects) or on day 60 (for the STT project), a laparotomy was carried out to examine the development of endometriosis in the animals. The total weight, area, and volume of the developed vesicles (lesions) were obtained, followed by an average per group of the totals from each animal. Various tissues were collected and stored as fresh frozen or fixed in 10% formalin71900-429670 for future processing, this included ovaries, uterus, distal colon, liver, adrenal glands, and brain. The colon was measured as described in the macroscopic scoring and stored in 10% formalin.Behavioral assessment of pain

[0072] The formalin test is a widely accepted measure of pain resulting from injury and involves a nociception of the inflammatory, neurogenic, and central mechanisms. One day before sacrifice, animals were brought into the testing room and allowed to acclimate for 10 minutes. Subsequently, a 2.5% formalin solution (50 pL) was injected into the right plantar hind paw of each rat.

[0073] Immediately following injection, each rat was placed into an empty, transparent cage, and its behaviors were recorded via video for one hour as the formalin test exhibits both an early and late phase. A blind observer quantified behaviors in blocks of 15 seconds at 5- rmnute intervals. A scoring scale based on the following criteria was used: "0" if the injected paw was not favored; "1" if the injected paw bears little or no weight; "2" if the injected paw was elevated and not in contact with any surfaces; and "3" if the injected paw was licked, bitten, or shaken. The formalin test was conducted only once at the conclusion of the study, with comparisons of experimental groups made against control and sham groups. The composite pain score for the formalin test was calculated using the formula: CPS = [(1 * category 2) + (2 * category 3)] / test duration, where the test duration is equal to 60 minutes. The early phase of the test (10-25 minutes) was used to correlate with macroscopic scoring. While experimental groups underwent other pain assessments, including the Von Frey and the Hot Plate tests, only the formalin is illustrated as it was used to correlate with macroscopic scoring.Macroscopic scoring

[0074] The colon of each animal was excised and subjected to macroscopic assessment by an observer that was blinded to treatment, employing a set of four pre-estabhshed criteria. Briefly, the blinded investigator recorded the existence of adhesions (e.g., graded as 0, 1, or 2, denoting the absence, minor presence, or significant presence of adhesions, respectively) and the manifestation of diarrhea (indicated as 0 or 1, representing the absence or presence of diarrhea, respectively).

[0075] Furthermore, a digital caliper quantified the colon wall thickness in millimeters. The mucosal surface of the colon was carefully observed to detect instances of inflammation and ulceration (scored as 0 for no impairment, escalating linearly to 10 based on the extent of hyperemia, inflammation, and ulceration). These individual assessments were cumulatively aggregated to yield an overall damage score, with a possible maximum score of 14 to 15 points71900-429670 contingent upon the thickness of the colon. Rats in the TAP and ENT protocols were not quantified for mucosal surface changes. Rats in the STT protocol were assessed for mucosal surface changes.Connective tissue staining

[0076] Paraffin-embedded sections of endometriosis vesicles were cut at 4 pm and collected on positively charged slides. The tissues were stained using a Trichrome staining kit (ab 150686, Abeam, Cambridge. UK). All materials and prepared reagents were gently shaken and allowed to equilibrate at room temperature. Sections were deparaffinized and placed in distilled water. Bouin's Fluid was pre-heated in a water bath to a temperature between 56-64 °C in a fume hood or well-ventilated area. The slides were submerged in the preheated Bouin's Fluid for 60 minutes and then cooled for 10 minutes afterward. Slides were rinsed in tap water until the sections became completely clear, followed by rinsing in distilled water. Equal amounts of Weigerf s (A) and Weigerfs (B) were combined, and the working Weigerfs Iron Hematoxylin solution was applied to the slide for 5 minutes, followed by a wash in tap water. Biebrich Scarlet / Acid Fuchsin Solution was applied to the slides and left for 15 minutes, then rinsed in distilled water. Slides were immersed in Phosphomolybdic / Phosphotungstic Acid Solution for 10-15 minutes or until the collagen was no longer red, immediately followed by Aniline Blue Solution for 5-10 minutes. An additional rinse in distilled water was followed by a 1% acetic acid solution on the slides for 3-5 minutes. Slides were dehydrated, cleared in xylene, and cover-slipped.Microscopic quantification of connective tissue

[0077] Slides were imaged at 40x on an Olympus BX-60 (Corporate Parkway, NJ) equipped with a Nikon DS-F11 camera. A semiquantitative grading system for endometriotic vesicles was adapted. First, the collagen deposition (fibrosis) staining pattern was scored on a scale of 0 to 3. The scale was as follows: 0, no increase showing perivascular collagen only (normal); 1, slight increase with focal collagen deposition without interconnections; 2, moderate staining show ing focal interconnections of tick collagen bundles; and 3, marked staining showing diffuse deposition of interconnecting collagen bundles. If the pattern was heterogeneous, the final score was determined by the highest score observed in at least 30% of the tissue section. The second parameter was the collagen deposition extent and involvement, estimated as the percentage of collagen fibrosis (blue staining areas) per tissue section examined. The scoring per section was as follows: 0= <1%; 1= 1-25%; 2= 26-50%; 3= 51-75%; 4= >75%. The final grading of fibrosis was calculated by adding the scores for the staining71900-429670 pattern of collagen and the percentage of tissue involved. The fibrosis grading score ranges from 0 to 7.EXAMPLE 2Endometriosis Development

[0078] On day 28, after surgical induction of endometriosis, a laparotomy was carried out on the ato examine the development of endometriosis in the animals. FIG. 2 summarizes the parameters quantified for endometriosis vesicles for the TAP and ENT projects. FIG. 3 summarizes the results of the STT project. The total weight, area, or volume of the developed vesicles were evaluated, and then the average per group of the totals from each animal was calculated.

[0079] In FIGS. 2A-2C, pexacerfont did not change the percentage of developed vesicles, regardless of dose. Similarly, pexacerfont did not change the total vesicle weight (FIGS. 2D-2F) and total area of the vesicles (FIGS. 2G-2I) compared to vehicle control groups within each treatment.

[0080] For the STT project (FIG. 3), the percentage of developed vesicles ranged from 68% to 80% on average per group. However, there were no significant differences due to treatment (FIG. 3 A). Vesicle weight was lowest for the group of rats that received chronic pexacerfont (0. 13 ± 0.03 g SEM) and highest for the group that received chronic elagolix (0.26 ± 0.09 g). Still, ANOVA revealed no significant differences across groups (FIG. 3B). Changes produced by pexacerfont for the total vesicle area showed minimal changes across groups (FIG. 3C).EXAMPLE 3Macroscopic Scoring

[0081] FIG. 4 presents the quantification of adhesions, diarrhea, and colonic thickness for animals undergoing the TAP protocol (FIG. 4A) and the EnTrust protocol (FIG. 4B and FIG. 4C). For FIG. 4A and FIG. 4B, hyperemia was not quantified.

[0082] At 28 days after endometriosis progression, a dose of 3 mg / kg pexacerfont did not show- protection against the adhesions, diarrhea, or colon thickness, but at 10 mg / kg, protection became evident (FIG. 4C; t= 2.77, d.f = 22, p= 0.011). Without being bound by any theory, the result could be due to a larger magnitude decrease in adhesions for the 10 mg / kg dose (32% change for vehicle vs. pexacerfont) as compared to the 3 mg / kg dose (20% change71900-429670 vehicle vs. pexacerfont; see Table 1). When the same measures were taken after seven days of endometriosis progression, there was no significant difference due to treatment, mainly because the progression of the endometriosis is shorter and there is less damage in general. The total score in the vehicle group of FIG. 4B (1.58 ± 0.21) is half that in the vehicle group of FIG. 4C (2.46 ± 0.20).Table 1. Macroscopic scoring and individual values

[0083] Macroscopic score findings in the STT project (FIG. 5) show a significant main effect of treatment as revealed by ANOVA (F(5,35)= 11.76, pO.OOOl). Post hoc analyses showed that endometriosis increased macroscopic scores over sham controls (vehicle-sham vs. vehicle-endometriosis, p< 0.0001). The increase in macroscopic score was controlled with both acute and chronic pexacerfont administration at 10 mg / kg as evidenced by the average of both groups being lower than the vehicle-endometriosis group (p< 0.01 for both groups).

[0084] In comparison, elagolix (a common therapeutic to treat endometriosis) did not demonstrate protective effects for macroscopic scoring in animals (p= 0.335 compared to the vehicle-endometriosis group). Evaluating each component of the macroscopic score in Table 2 and in FIG. 5 demonstrates that adhesions were 37% and 51% less in the pexacerfont acute administration and the pexacerfont chronic administration groups compared to the endo-vehicle group. Colons observed in rats with endometriosis demonstrated decreased thickness in response to acute and chronic administration of pexacerfont compared to endo-vehicle, showing71900-429670 a 34% and 28% decrease, respectively. Diarrhea was seldom observed, while changes in hyperemia were more variable.

[0085] To evaluate if CRH1 receptor antagonists decrease the overall incidence of adhesions, the percentage of animals that developed any type of adhesions was quantified compared to those that received a score of zero (no adhesions). For the animals that received 3 mg / kg pexacerfont immediately after surgery and assessed after 28 days of surgery, 100% of the rats in the vehicle groups developed adhesions, compared to 91.6% of the rats in the pexacerfont group [RR: 0.48 (95% CI= 0.29 to 2.40)].

[0086] When a pexacerfont dose of 10 mg / kg was administered followed by assessment at 28 days, 100% of the rats in the vehicle group developed adhesions, and 83.3% of rats receiving pexacerfont demonstrated adhesions [RR: 0.45 (95% CI= 0.27 to 1.42)]. Assessment at 7 days after surgery (i.e., when pexacerfont (10 mg / kg) administration ended) resulted in 66.6% of the rats in the pexacerfont group displaying adhesions compared to 91.6% in the vehicle group [RR: 0.61 (95% CI= 0.37 to 1.16)]. Therefore, the risk of developing adhesions following administration of pexacerfont after surgery was decreased by 39-55% in animals, depending upon the dose and time assessed (calculated as 1-RR).

[0087] For the STT protocol, administration of pexacerfont began 25 days after endometriosis-induction surgery and adhesions were assessed 60 days after surgery. Some development of adhesions had already occurred. Thus, pexacerfont administration was evaluated as a therapeutic agent for reduction of prevention of further adhesion formation.

[0088] For animals with endometriosis that received either vehicle or elagolix, 100% of the rats had adhesions after 60 days. However, rats receiving pexacerfont as acute administration and chronic administration showed 92.3% and 71.4% of animals with adhesions,71900-429670 respectively (FIG. 6). This reduction between 7.7 and 28.6% in the incidence of adhesions occurred even when the treatment was started significantly later after the surgical event. Only- one rat showed mild adhesions in the sham group that received the vehicle, representing 9% of the sample.EXAMPLE 4Reactive Responses to Inflammatory Pain

[0089] To understand the effects of endometriosis and the therapeutic on inflammatory pain responses, the CPS was quantified during the pain onset of animals (between 10-25 minutes) and the pain offset of animals (between 30-55 minutes). FIG. 7A illustrates the normalized CPS during pain onset for the sham vehicle response. Animals with endometriosis receiving chronic administration of pexacerfont or elagolix showed a decrease in pain responses (negative values), which is suggestive of analgesia.

[0090] One sample t-test demonstrated that animals with endometriosis receiving chronic administration of pexacerfont had a significant average reduction in CPS (t=3.73, d.f = 6, p=0.009, uncorrected. p=0.039 Bonferroni corrected). Other groups did not demonstrate a significant difference from the baseline. Simple linear regressions per group were performed to evaluate understand association between inflammatory pain and macroscopic scores (FIG. 7B). While none of the linear regressions differed from zero, endometriosis appears to cause a positive regression slope between CPS and macroscopic score that is not observed in animals without endometriosis (sham vehicle group). Animals administered elagolix demonstrated a similar positive regression as the endometriosis-vehicle group. However, acute administration of pexacerfont reversed the positive regression slope to a negative regression, similar to observation in the sham vehicle group. Chronic administration of elagolix in rats wi th endometriosis showed a nearly flat regression. Therefore, acute administration of pexacerfont abolished the association of macroscopic score and inflammatory pain responses, resulting in a similar effect to the group without endometriosis.EXAMPLE 5 Microscopic Scoring

[0091] The instant example provides microscopic scoring results for the TAP protocol. Trichrome analysis of endometriotic vesicles from animals that were administered 3 mg / kg of pexacerfont showed a 31% decrease in the histological grading as compared to the vehicle71900-429670 group (pexacerfont: 1.75 ± 0.40; vehicle 2.52 ± 0.36; t=1.43, d.f.=17.20, p=0.17). These data suggest evidence that pexacerfont decreases inflammatory pain responses along with a decrease in colonic macroscopic score (with and without quantified hyperemia) as an indicator of peritoneal-colonic damage produced by the progression of endometriosis. Specifically, these results show a consistent reduction in adhesions, a known complication of endometriosis development. Thus, pexacerfont administration can be effective against endometriosis- associated GI damageEXAMPLE 6Materials and Methods for Examples 7-13Animals

[0092] Female Sprague Dawley Rats, aged 60 days and with an average weight of 180 grams, were procured from the Animal House at Ponce Research Institute (PHSU, Ponce, Puerto Rico). Rats were housed in pairs in a controlled environment with a room temperature of 23 °C and a 12-hour light-dark cy cle. Standard laboratory chow and tap water were provided ad libitum, and bedding was changed twice weekly. To minimize experimenter-induced stress, rats were handled for 7 days before endometriosis induction. Additionally, vaginal smears were collected to monitor the regularity' of the estrous cycle. Experiments were conducted in the morning to minimize the influence of circadian rhythms. Animals were randomly allocated to five treatment groups: endo-vehicle acute administration (n=15), endo-pexacerfont acute administration (n=13), endo-pexacerfont chronic administration (n=l 1), endo-elagolix chronic administration (n=l l), and sham-vehicle acute (n=l l).Endometriosis auto transplantation surgery induction

[0093] The surgical procedures for inducing endometriosis in rats were conducted following established protocols. Rats w ere initially anesthetized in a chamber with 4% isoflurane and maintained at 2-3% using an open mask once positioned supine over a w arm pad to maintain adequate body temperature. Under sterile conditions, a midline laparotomy was performed to expose the right uterine horn. A 2 cm segment of the distal portion was excised and placed in a sterile culture medium. Subsequently, the endometrium was exposed by carefully opening the right uterine hom, and four pieces measuring 2 mm x 2 mm w ere excised. These endometrial implants were sutured onto the mesentery of the small intestine, w ith the serosal surface facing the mesentery and positioned adjacent to the vessels.71900-429670

[0094] For the sham procedure, rats underwent a simulated surgery in which the right uterine hom was gently massaged for 2 minutes with fingertips to mimic mechanical handling without implantation. Additionally, four sutures were placed on the mesentery without the inclusion of uterine implants. The peritoneal cavity was irrigated throughout the surgery with copious amounts of sterile saline solution to minimize adhesions. Consistent with previous research protocols, endometriosis was allow ed to progress for a total of 60 days before rats w ere euthanized.Behavioral assessments

[0095] Before surgery, baseline assessments of mechanical and thermal pain w ere conducted in the rats (FIG. 8). Following the induction of endometriosis but before therapy administration, assessments were repeated. An electronic Von Frey (DCA Software Ugo Basile) was utilized to evaluate mechanical pain or allodynia. Rats were acclimatized for 15 minutes in separate compartments (17cm x 69cm x 14cm), followed by an additional 15 minutes of acclimatization w ith the investigator. A vinyl filament was applied to the left hind paw of each rat three times, follow ed by three applications to the right hind paw. Force and time were given by the apparatus and recorded.

[0096] A hot plate analgesia meter was employed to assess thermal pain or hyperalgesia. Each rat was acclimatized at room temperature on the hot plate apparatus for 5 minutes prior to testing. The plate was preheated to 52 °C before use. Animals were placed on the hot plate for 40 seconds, and the rat was recorded the whole time. Reaction time was later analyzed from the video recordings.

[0097] Evoked pain due to injury (involving inflammatory, neurogenic, and central mechanisms of nociception) w as measured using the formalin test. This test w as conducted only once, on the day prior to euthanasia. A 2.5% formalin solution was prepared by diluting 3 mL of water with 1 mL of 10% neutral buffered formalin. Each rat received a 0.05 mL injection of the formalin solution into the right hind paw. Animals were placed in individual modules and recorded for one hour. For analysis, the recorded videos were used for scoring as follows: a blinded observer quantified behaviors by watching the video every 15 seconds for 20 observations in a span of 5 minutes to obtain a score of the most repeated behavior in that 5- minute block. This was repeated every 5 minutes for the duration of the video. The scoring scale is based on the following criteria: "0" if the injected paw is not favored, "1" if the injected paw7has little or no weight, "2" if the injected paw7is elevated and is not in contact with any surfaces, and "3" if the injected paw is licked, bitten, or shaken. The sum of each of the 12 scores was obtained and the CPS formula used for statistical comparison between treatments.71900-429670Therapeutic Administration

[0098] Food-grade methylcellulose 0.5% was diluted in water and served as the vehicle for the experiment. Animals receiving pexacerfont. a corticotropin-releasing hormone antagonist, were administered at a concentration of 10 mg / mL, diluted with the 0.5% methylcellulose solution. Elagolix, a commonly used hormonal therapeutic, was employed as a positive control at a concentration of 14 mg / mL, diluted in 0.5% methylcellulose in PBS. Pexacerfont was purchased primarily from Biotechne R&D Systems (Minneapolis, MN. USA), and subsequently from Biosynth International, Inc. (San Diego, CA, USA). Elagolix was purchased from MedChem Express (Cat. No.: HY-14789; Monmouth Junction, NJ, USA).

[0099] Rats undergoing acute treatment were administered the therapeutics starting 25 days after endometriosis induction and continued for seven consecutive days. Chronic treatment regimens were initiated on days 25, 39, and 53 post-endometriosis induction (see FIG. 8). All treatments w ere orally administered to the animals by placing the drug suspension or vehicle solution inside oyster crackers. Complete ingestion of the crackers was observed in each of the rats, indicating full adherence to the treatments. Gavage was not used to avoid repeated stress to the animal, which constitutes a confounding factor with endometriosis development.Smears

[0100] The regular cycling pattern of the rats was confirmed by analyzing morphological changes indicative of the different phases of the estrous cycle (diestrus. proestrus, estrus, and metestrus) over several days. Vaginal smears were collected seven days prior to endometriosis induction to ensure baseline regularity in cycling patterns. On the day of surgery' and during behavioral assessments, rats underwent vaginal cytological smears to verity' consistent estrous cyclicity’. Similarly, on days of therapeutic administration, smears were obtained to assess the treatments' impact on reproductive cyclicity, if any. Finally, all animals underwent vaginal smear analysis on the day of sacrifice to evaluate estrous cycle phases and their potential influence on experimental outcomes.Collection of Tissues and Endometriosis Severity

[0101] After 60 days of endometriosis progression, animals were euthanized to evaluate disease severity and to collect tissues. A one (1 mL solution of pentobarbital (65 mg / kg) w as administered intraperitoneally to each rat before laparotomy. Upon verification of the appropriate plane of anesthesia, a terminal laparotomy was conducted to assess the extent of endometriosis and retrieve relevant tissues. Each was meticulously examined for the presence71900-429670 of implants and the original sutures. Measurements of vesicles longest length and width were obtained using a digital caliper, and their growth was graded based on a modified scoring system. Briefly, vesicles <2 mm in length were assigned a score of 2, while those > 2 mm and < 4.5 mm received a grade of 3. Vesicles measuring > 4.5 mm and < 6.0 mm were graded as 4, and those > 6.0 mm were graded as 5. Implants that had disappeared were graded as 1.

[0102] The entire colon of the animals w as excised and examined for macroscopic damage, including the presence of adhesions, ulceration, diarrhea, and colon thickness, utilizing a previously established scoring system. Tissue segments were weighed and fixed in 10% formalin. Vesicles w ere rapidly frozen in liquid nitrogen, and serum and peritoneal fluid w ere collected and stored at -80 °C until further assay.Histology in Vesicles and Uteri

[0103] After processing the tissue, vesicles, and uteri tissue segments of 4 pm were stained with hematoxylin and eosin to determine glandular epithelium and collagen deposition.Immunohistochemistry

[0104] Formalin-fixed and paraffin-embedded endometriotic vesicle samples were cut at 4 pm thickness with a microtome (Microm HM340 E, Microm International) and mounted on positively charged glass slides. Tissue sections were deparaffinized with xylene, tw o changes, 15 minutes each, and hydrated in descending grades of ethanol for 3 minutes each to distilled water for 1 minute. This was followed by a 3% hydrogen peroxide (Sigma-Aldrich) incubation for 15 minutes to block endogenous peroxidase and a 5-minute PBS wash. After antigen retrieval (0.01 M Citrate-EDTA buffer, pH 6.0, 95-99 °C for 40 minutes), slides were cooled for 20 minutes at room temperature, rinsed with tw o changes of distilled water for 2 minutes, and placed in PBS for 5 minutes. Slides were blocked with normal serum (HK112-9K Bio Genex) for 15 minutes and followed by an overnight incubation with primary antibody (rabbit polyclonal NGF antibody catalog # sc-548 Santa Cruz Biotechnology, dilution 1 : 100, VEGF antibody catalog # sc-7269 Santa Cruz Biotechnology', Inc, dilution 1:50 used respectively). A negative control with PBS instead of the primary antibody was run on each slide. On the second day, slides were washed with PBS for 5 minutes. A multi-link was used as the secondaryantibody for 20 minutes, followed by PBS wash for 5 minutes. The slides were incubated with Streptavidin Peroxidase for 20 minutes (LP000-ULE Bio Genex). For development, one drop of 3,3' Diaminobenzidine (DAB) (HK153-5KE Bio Genex) was used on each tissue, and the exposure was monitored for 3 minutes under a light microscope. Then, the slides were dipped in distilled water, counterstained with hematoxylin for 15 seconds, washed with running water for71900-4296705 minutes, dehydrated through graded alcohol, cleared with xylene, and mounted with cytoseal 60 (Epredia, Cat # 23-244256 Fisher Scientific). Four distinct areas were photographed and evaluated independently by three blinded observers. Staining intensity was graded using a scoring system ranging from 0 (no staining) to 3 (strongest staining). Additionally, each area was assessed for the extent of staining: 0 for 0-10% of tissue stained, 1 for 10-40%, 2 for 40- 70%, and 3 for 70-100%.Immunofluorescence

[0105] Ki-67, a cell marker for proliferation, was used on segmented endometriotic vesicles cut at 4 pm. Briefly, formalin-fixed paraffin-embedded tissue sections were deparaffinized with xylene-substitute, hydrated with descending grades of alcohols, boiled in Citrate-EDTA for 40 minutes, and blocked with normal goat serum. Tissues were incubated with primary antibodies (Ki-67 1 :50 dilution, cat#550609 BD Pharmingen) overnight at 4 °C in a humidifying chamber. After washing with PBS, the tissues were incubated with highly crossabsorbed secondary antibodies (Molecular Probes by Thermo Fischer Scientific, Inc.), for 30 minutes at room temperature in a humidifying chamber. All tissues were counterstained with a nuclear dye, DAPI (R37606 Invitrogen by Thermo Fisher Scientific), for 5 minutes. A tissue section on each slide was used as a negative control, receiving PBS instead of the primary antibody. Tissues were visualized with Nikon Confocal Microscopy. The integrated densify of three representative areas for each vesicle tissue was analyzed using ImageJ Software.Serum Hormone Levels

[0106] The Milliplex Rat Pituitary Magnetic Bead Panel (RPTMAG86K, MilliporeSigma) was used according to the manufacturer’s protocol to measure the concentration of Luteinizing Hormone (LH) and Follicle-Stimulating Hormone (FSH). All samples were run in duplicate in the Luminex MAGPIX system, and concentrations were expressed in pg / ml.Gonadotropin-Releasing Hormone ELISA

[0107] Rat (GnRH) Gonadotropin-Releasing Hormone ELISA Kit (EKF57934, BIOMATIK) has high sensitivity and excellent specificity for detecting GnRH. GnRH was measured in the rat serum in a 1 : 10 dilution following the manufacturer’s protocol. All samples were run in duplicate in the MultiSkan Reader, and concentrations were expressed in pg / mL.Cytokines PCR71900-429670

[0108] Vesicles frozen in liquid nitrogen were used for RNA extraction. Thirty milligrams of tissue were weighed and transferred to mRNAse-free micro centrifuge tubes filled with beads and homogenizing solution. Tissue samples were homogenized for 5 minutes using the Bullet blender (Advance Co. Troy, NY). Samples were extracted using RNeasy Mini Kit (QIAGEN Cat. No.: 74106). After extraction, RNA concentration and quality were verified using Nanodrop 2000 (Thermo Scientific Cat No.: ND2000). Then, 1 pg from the mRNA was converted to complementary DNA using iScript cDNA synthesis kit (BioRad Cat. No.: 1708891BUN). qPCR was performed using iQ SYBR Green Supermix (cat # 1708882, BioRad) and primers for IL-1 (3 (QIAGEN Cat. #:330001- PPR06480B-200), IL-6 (cat # 330001- PPR06483B-200 Qiagen) and TNF-a (Cat. #: 330001 -PPH00341F-200 Qiagen). Data were reported as fold change using the equation 2-ACT.EXAMPLE 7 Vesicle Lesion Sizes

[0109] At sacrifice, none of the sham-operated animals developed vesicles at the suture sites. In contrast, all rats with induced endometriosis developed vesicles at least at one implant site. The percent of developed vesicles was calculated as [(number developed vesicles / total number implants) x 100%] and then averaged per treatment group. Vesicles developed at 80% of suture sites among the endometriosis vehicle-treated group. Non-parametric Kruskal-Wallis test showed no difference in the percentage of vesicle development among the drug-treated groups compared to the vehicle group [FIG. 9A; H(4)= 0.814, p> 0.05], The total weight and area of developed vesicles was also measured per animal and averaged within the treatment groups (FIG. 9B, FIG. 9C). One-way ANOVA showed no significant differences in vesicle weight [F(3,27)= 0.48, p> 0.1)] or area [F(3,38)=0.07, p>0. 1] between the experimental groups. The grading of vesicles based on their length is shown in FIG. 9D. Animals in the endometriosis vehicle group exhibited vesicles sized 6.0 mm or larger (grade 5) in 15% of cases. In contrast, those in the endometriosis pexacerfont acute group showed this in only 1% of cases. There was a trend towards lower grade 3 vesicles for the endometriosis pexacerfont acute and endometriosis elagolix chronic groups compared to endometriosis vehicle.EXAMPLE 8Pexacerfont Decreases NGF, VEGF, and Ki67 Within Vesicles71900-429670

[0110] A semi-quantitative analysis of the staining area and intensity for NGF and VEGF in formalin-fixed endometriosis vesicles was performed using immunohistochemistry. For VEGF, groups with endometriosis receiving acute and chronic pexacerfont showed significantly lower VEGF intensity than those in the endometriosis vehicle group [FIG. 10A; F(3,14.7)= 8.48, p< 0.01; Dunnett’s: p< 0.01 and p=0.05, respectively). Similarly, the VEGF area was significantly lower in the endometriosis pexacerfont acute and chronic groups compared to the endometriosis vehicle group [FIG. 10B; F(3.16.2)= 8.43, p< 0.01; Dunnett's p< 0.01 and p=0.05 respectively]. The endometriosis group that received elagolix was not different from the vehicle control group (p> 0. 1 for both intensity’ and area).

[0111] A significant decreases in staining intensity and area was observed with acute pexacerfont for NGF (FIG. 10C, FIG. 10D). ANOVA followed by Dunnett’s post hoc showed a significant main effect for intensity [F(3,7.8)= 10.23, p< 0.01] and area [F(3.8.5)= 10.48, p< 0.01], with the respective post hoc test revealing a difference between acute pexacerfont group and vehicle for both measures (p< 0.01 in both intensity and area). However, no differences compared to the vehicle group were noted in the chronic therapeutic treatment groups (p> 0.1).

[0112] Using immunofluorescence, the integrated density (area x intensity) for Ki-67 was also quantified as a cell proliferation marker. Analysis revealed that endometriosis animals that received chronic pexacerfont had a significantly lower Ki-67 density compared to the endometriosis vehicle group [F(4, 12. 1)= 6.22, p< 0.01; Dunnett’s multiple comparison, p< 0.05], While the endometriosis group that received elagolix also showed a decrease in Ki-67, this difference did not reach statistical significance (p= 0.06).EXAMPLE 9Pexacerfont Decreases Allodynia and Inflammatory Pain

[0113] Using the Von Frey test, allodynia was measured at three different time points during the experiment: before endometriosis induction (baseline), before the first therapeutic administration (Day 24, pre-drug), and before sacrifice (Day 59, pre-sacrifice). A two-way ANOVA analysis of force normalized to sham comparison found a statistically significant difference in endo-pexacerfont acute at pre-surgery vs. pre-sacrifice [Main effect of time: F(2,l 16)= 23.24, p< 0.001, post hoc, p <0.01] and pre-drug vs. pre-sacrifice (p< 0.01). In endo- pexa chronic and endo-elagolix chronic, a significant difference in pre-drug vs. pre-sacrifice (p< 0.05 and p<0.01, respectively) was seen. These results show that animals that received pexacerfont and elagolix chronically resist more force and, thus, have higher pain thresholds and less allodynia (FIG. 11 A).71900-429670

[0114] Using the Hot Plate test, thermal pain or hyperalgesia was measured. A two-way ANOVA analysis of the latency to withdraw the forepaw revealed a main effect of time [F(2, 112)= 3.19, p< 0.05], Still, post hoc analyses showed no difference among groups (FIG.1 IB). Inflammatory pain was measured only once on day 59 of the experiment, right before sacrifice, using the formalin test. An ordinary one-way ANOVA showed a statistically significant difference in the composite pain score (CPS) change using the sham group as the baseline [F(3,46)= 4.36, p< 0.01], Specifically, the endo- pexacerfont chronic group showed significantly lower CPS compared to the endo vehicle (Dunnetfs post-hoc, p< 0.05). However, none of the other treatment groups showed significant differences from vehicle-treated animals (FIG. 11C).

[0115] Alleviation of pain is one of the primary clinical goals for endometriosis. Nevertheless, pain management in this clinical population is significantly complicated by other types of pain, such as bladder pain syndrome, pelvic myalgia, and vulvodynia, among others. Pain management in endometriosis also needs to consider neural mechanisms that include peripheral, central, and cross-sensitization. Therefore, focusing on signaling cascades that may impact all these factors in the peripheral and central nervous system is desirable. CRH is uniquely positioned to work both at the central and peripheral (paracrine) sites, including the reproductive tissues. CRH serves as the principal regulator of stress by activating the hypothalamic-pituitary-adrenal (HP A) axis. Beyond this established pathway, it is documented that chronic pain induces disruptions in limbic structures, such as the amygdala and hippocampus, through CRH activity. Conversely, it is recognized that the paracrine activity of CRH in uterine tissue contributes to endometrial inflammatory responses, and the activation of CRH can be stimulated by prostaglandins. Central and peripheral CRH activities signal painful stimuli. Despite the function of CRH in pain modulation, these examples demonstrate an analgesic effect in an endometriosis model.EXAMPLE 10Pexacerfont Does Not Impact Gonadal Hormone Cascade or Estrous Cyclicity

[0116] To assess if the therapeutic treatment altered gonadal hormones, the estrous cycle before surgical induction of endometriosis, during the first therapeutic treatment week (days 25-31), and during the subsequent w eeks of therapeutic administration in groups with chronic therapeutic treatment (days 39-45 and 53-59) was monitored. All animals had normal estrous cyclicity prior to surgery (FIG. 12A); animals also continued to cycle normally regardless of the treatment. Serum GnRH, LH, and FSH levels were also quantified on the day71900-429670 of sacrifice. GnRH levels were unaffected by treatments (FIG. 12B). However, when the FSH / LH ratio was calculated (samples taken in the morning), there was a significant decrease in the hormonal ratio in rats that received elagolix compared to the vehicle group (FIG. 12C; dark symbols represent rats in estrus). Non -parametric analysis showed a mam effect of treatment [H(5)= 14.4, p< 0.01, Dunns post hoc p< 0.05], This decrease in the hormonal ratio was not observed for any of the other treatment groups in comparison to vehicle treated group.EXAMPLE 11Pexacerfont Significantly Decreases Macroscopic Score

[0117] Macroscopic score assessment included adhesions, diarrhea, colon thickness, and colon hyperemia at the time of sacrifice. Statistical analyses revealed a significant main effect of treatment [Table 1; F(5,35)= 11.76, pO.OOOl], Post hoc analyses showed that endometriosis increased macroscopic scores over sham controls (vehicle-sham vs. vehicle-endometriosis, p< 0.0001). The increase in macroscopic score was controlled with both acute and chronic pexacerfont since both groups' averages were lower than the vehicle-endometriosis group (p< 0.01 for both groups). The endo-elagolix group also showed a decrease in the macroscopic score compared to endo- vehicle (p< 0.05), but of weaker magnitude than the endo- pexacerfont chronic group. When each component of the macroscopic score in the table shown in Table 3, was observed, it can be seen that adhesion scores were 37% and 51% less in the endo- pexacerfont acute and endo- pexacerfont chronic compared to the endo-vehicle group. The colon in rats with endometriosis showed decreased thickness in response to acute and chronic pexacerfont compared to endo- vehicle of 34% and 28%, respectively. Diarrhea was seldom observed, while changes in hyperemia were more variable.Table 3. Macroscopic scores71900-429670

[0118] The finding that pexacerfont reduces macroscopic score, particularly peritoneal adhesions, is clinically significant for treating endometriosis. Adhesions can develop when endometrial lesions bleed into the surrounding tissue, triggering an inflammatory response that forms bands of tissue that connect two organs. Reports show that adhesions associated with endometriosis could present as thin, filmy, and transparent to thick, dense, and opaque. In severe cases, pelvic adhesions can lead to a potentially life-threatening condition known as "frozen pelvis". Moreover, the quality of life in infertile women with endometriosis-related adhesions cab be significantly impacted. Up to 74% of women present endometriosis-associated adhesions before any surgical intervention, and repeated surgical interventions significantly exacerbate this problem. No current oral pharmacological treatment is on the market for mitigating endometriosis-associated adhesions. The presence of dense adhesions in endometriosis patients is associated with increased severe complications, such as ureteral and rectal injuries, as well as voiding dysfunction. Directly targeting adhesions in endometriosis could lead to decreased laparoscopic complications, resulting in increased savings in direct healthcare costs.EXAMPLE 12Pexacerfont Decreases IL-6 and TNF-alpha mRNA Within Vesicles

[0119] Using qRT-PCR, three inflammatory cytokines frequently reported in patients with endometriosis were quantified: IL-6, IL- 1(3, and TNF-a (see FIGS. 13A, 13B, and 13C). Non-parametric statistical comparisons revealed a main difference between groups [H(4)= 11.52, p< 0.01], with the endo- pexacerfont chronic and the endo- elagolix chronic showing a significant decrease compared to vehicle (Dunn’s post hoc, p< 0.05 for both comparisons). No statistical differences w ere observed for I L- 113. However, TNF-a show ed a similar pattern to that observed for IL-6, where groups with chronic pexacerfont and elagolix administration had a decrease in mRNA expression within the vesicles [H(4)= 12.64, p< 0.01; post hoc. p= 0.05 and p< 0.002 respectively]. The same cytokines were also quantified in peritoneal fluid using a71900-429670 multiplex plate to observe changes in the peritoneal environment (see FIG. 14A and FIG. 14B). No significant changes in IL-6 were noted. However, the expression of IL-1 P showed a difference between groups [F(5,27)= 2.70, p< 0.05], with only the endo-vehicle group showing a significantly increased expression over the sham group (post hoc. p< 0.05). TNF-a levels were undetectable in the peritoneal fluid samples and, thus, not shown.EXAMPLE 13Pexacerfont Preserved the Leptin-to-Body Weight Relationship in Animals

[0120] Using the multiplex, leptin in the rats’ peritoneal fluid was measured, which has been reported to play a role in endometriosis development. It is well described that increased fat content results in increased leptin release. Using the animal’s body weight as a proxy for increased fat, the relationship between peritoneal leptin and body weight at sacrifice was analyzed for all the treatment groups (FIG. 15A-FIG. 15D). There were no significant differences in peritoneal leptin levels among groups (FIG. 15 A). Similarly, the rats’ body w eight at sacrifice was comparable among groups, averaging 271 across all groups (FIG. 15B).

[0121] However, when a simple linear regression analysis between body weight and peritoneal leptin levels was used, a negative correlation for the endo-elagolix endometriosis group (slope= -0.16) w'as observed compared to all other groups (Sham= 0.38, endo-vehicle= 0.28, endo- pexacerfont acute= 0.15, endo- pexacerfont chronic= 0.45). This suggests a deregulation of the hormonal peritoneal environment resulting from elagolix treatment (FIG. 15C, and FIG. 15D).

[0122] Pexacerfont had a negligible impact on the gonadal axis as measured by a lack of effect on the estrous cycle and no alterations in the FSH / LH ratio. Low' FSH / LH ratios in the clinical setting (less than 1.34), as observed for elagolix herein, are associated with an increase in follicular phase length (+2.4 days) and a lower ovulatory rate, without changes in luteal phase length nor progesterone levels. Parallel to this, leptin has been established as a potent modulator of fertility in animals and humans. These experiments show ed an expected positive correlation of peritoneal leptin levels with higher animal body weights. However, the group that received elagolix showed a completely reversed linear correlation with a negative slope. While lower leptin might initially appear favorable for fertility, low leptin levels could also lead to impairments in immune function and irregularities in menstrual cycles. Taken together, targeting the CRH activity w ith pexacerfont appears as a safer alternative than the GnRH pathway as it shows minimal effects in the gonadal axis activity, which could represent a decrease in undesired side effects for the clinical scenario.71900-429670EXAMPLE 14Preventive Treatment Against Post-Surgical Abdominal Adhesions

[0123] A study was carried out using a surgical / cecal abrasion model on equally divided male and female Sprague-Dawley rats 8 to 10 weeks old, with an average weight of approximately 260 grams. Four treatment groups of rats were used: naive, abrasion / kaolin- vehicle, abrasion / kaolin-pexacerfont (3 mg / kg and 10 mg / kg). Batches of 16 animals at a time were assessed, with 4 batches in total, resulting in 15-16 animals per treatment. Animals were anesthetized with isoflurane and placed in a supine position to perform a laparotomy. The cecum was exposed, and a saline-soaked gauze was used to rub the cecum repeatedly for 54 seconds until subserosal bleeding occurred over an area of 1 cm2. Kaolin was used at 0.005 g / mL and 2 mL was applied with a syringe to the cecum, which causes consistent adhesions without compromising rat viability. The cecum was returned to the abdomen, and the walls sutured shut. Animals received pexacerfont orally starting the day after the model induction for 7 consecutive days, at which point the animals were sacrificed. The therapeutic suspension was measured based on the daily weight of the animal and injected into an oyster cracker.Assessment of pain behaviors using Von Frey filaments (VF: to measure mechanical allodynia) and hot plate testing (HP: to measure thermal analgesia) took place the day prior to the surgery and the day prior to sacrifice, as shown in the timeline exhibited in FIG. 17.

[0124] All animals were assessed for presence and severity of adhesions and peritoneal inflammation at the time of sacrifice (14 days) following the established protocols used routinely. To microscopically confirm the effects of the therapeutic in adhesion prevention, the intestinal caecum (exhibiting the presence of adhesions or not) was collected for histological analyses. After proper fixation, serial tissue sections were stained with H&E, and Masson's tri chrome to evaluate fibrosis formation. The team’s clinical pathologist scored tissues based on adaptation of a previously established scale for fibrosis formation. For detailed procedures refer to Examples 2-5 (endometriosis model). Mast cells in the distal colon and cecum were counted to assess inflammation. Tissue segments, distal colon and cecum, were fixed in 10% formalin by routine histological techniques, and mounted on glass slides then stained with Toluidine blue to visualize mast cells. Two observers randomly selected five high power fields (hpf) at 40x and an average number of mast cells per hpf was calculated.Weight change71900-429670

[0125] As shown in FIG. 18, the naive animals consistently gained weight throughout the experiment, while those receiving the surgical abrasion initially lost weight before recovering. There were no significant differences in the pattern of weight gain between surgical treatment groups in food consumption (not shown). Female animals weighed significantly less than their male counterparts at the time of sacrifice due to an overall slower growth rate and lower start weight. However, there were no significant differences between treatment groups within males or females.Behavioral assessment

[0126] FIG. 19A and FIG. 19B summarize the behavioral results. As noted, there were no significant differences in the mechanical allodynia or thermal hyperalgesia responses, and no differences between males and females. The closed symbols represent the males, while the open symbols represent the females. The first bar represents baseline behavior, and the second represents treatment effects right before sacrifice. Based on the vehicle treated group data, this model does not appear to cause chronic pain in the animals as no changes in allodynia or hyperalgesia were observed.Adhesion score

[0127] All animals were evaluated using a previously defined adhesion score where 0= no adhesions, 1= thin filmy adhesions, 2= more than one thin adhesion, 3= thick adhesion with one focal point, 4= thick adhesion with planar attachment, and 5= very thick vascularized adhesions or more than one planar adhesion. As shown in FIG. 20 A, there was significantly more development of thick or very thick vascularized adhesions (69%) in the vehicle-treated animals. Both doses of pexacerfont decreased this, with 50% of lesions found with the 10 mg / kg dose being categorized as thin and filmy or non-existent. This decrease in adhesions reached significance if the colon alone was considered (FIG. 20B, p<0.05). In this and subsequent graphs, individual data points are displayed, with full circles representing males and half-filled circles representing females.Cecum score

[0128] Having observed that the cecum of the animals was impacted in different ways from the colon which were not adequately reflected, an additional scoring system specifically for assessment of the cecal area was developed as follows: grade 0 = cecum was of normal size with no hyperemia, angiogenesis, or adhesions; grade 1 = cecum had mild inflammation with mild hyperemia, and adhesions but no angiogenesis; grade 2 = cecum had moderate71900-429670 inflammation with adhesions and moderate hyperemia and angiogenesis; grade 3 = cecum had severe inflammation with adhesions and severe hyperemia and angiogenesis. As shown in FIG. 21, the surgical abrasion model had a significantly higher cecum score than non-surgical naive animals, and this score was reduced with both doses of pexacerfont but did not reach significance. No differences were observed between males (solid symbols) and females (empty symbols).Macroscopic damage

[0129] A general macroscopic damage score was used to characterize the overall impact on the large intestine. This scoring comprised assessing the presence of hyperemia and inflammation, colon wall thickness with a digital caliper (an indicator of inflammation), diarrhea (indicated as 0 or 1, representing the absence or presence, respectively), and colonic adhesions (graded as 0, 1, or 2, denoting the absence, minor presence, or significant presence, respectively).Table 4. Macroscopic score breakdown

[0130] As shown in FIG. 22, the higher dose of pexacerfont was able to significantly reduce this score (p<0.01), being attributed to all components besides diarrhea (see Table 4). The therapeutic appears to be equally effective in both males and females.Microscopic damage

[0131] Microscopic damage within the colon was assessed using a previously published scoring system to assess changes at the microscopic level in both the colon and cecum. This scoring system assessed loss of mucosal architecture (from mild to severe 0,1, 2 or 3); cellular71900-429670 infiltration where 0=absent (no inflammatory infiltrate), 1 (in muscularis mucosae), 2 (in lamina propria / villi) and 3 (in serosa); muscle thickening 0 (muscle < ! of mucosal thickness), 1 (muscle is 1 / 2 -3 / 4 of mucosal thickness), 2 (muscle = mucosal thickness), or 3 (all muscle); goblet cell depletion either 0 (absent) or 1 (present); crypt abscess formation either 0 (absent) or 1 (present).

[0132] As show n in FIG. 23A and FIG. 23B, the vehicle animals had a higher microscopic score compared to naive, reaching significance in the colon (p<0.05; FIG. 23A). Neither dose of pexacerfont had any significant impact on this damage in the colon or cecum (FIG. 23B).Fibrosis Scoring

[0133] A Masson Tri chrome Stain Kit (ab 150686) was used for fibrosis grading in tissue sections of both colon and cecum, which were then evaluated in a blinded fashion by a clinical pathologist. This scoring system took into consideration collagen deposition staining pattern: 0 = no increase (perivascular collagen, fine fibers only), 1 = slight (focal collagen deposition without interconnection, and in cecum loosely arranged fibrils with numerous fibroblast nuclei), 2 = moderate (focal interconnection of thick collagen bundles; and compactly arranged collagen deposition in cecum), 3 = marked (diffuse deposition of interconnecting collagen bundles; with mature compact collagen, fewer fibroblast nuclei in cecum); extent of collagen deposition: 0 (<1% of section). 1 (1-25%), 2 (26-50%), 3 (51-75%), and 4 (>75%); vascular proliferation (blood vessels / hpf): 0 = no increase (0-1), 1 = slight (>I-5), 2 = moderate (>5-9), 3 = marked (>9); and neural tissue proliferation (nerve fibers / hpf): 0 = no increase (0-1), 1 = slight (>l-5), 2 = moderate (>5-9), and 3 = marked (>9). The summed scores are shown in FIG. 23A and FIG. 23B with colon (FIG. 23 A) and cecum (FIG. 23B). Cellular infiltration w as also assessed using an H&E stain, where 0 = absent, 1= slight, 2 = moderate, and 3 = marked. This was added to the fibrosis scoring. The sum of the five scores for each animal (total final grading score) is shown in FIG. 23C and FIG. 23D. There were no significant differences in scores noted across groups in the colon (FIG. 23C). In contrast, in the cecum (FIG. 23D) the pexacerfont groups tended to be higher (but not significant).Mast cell analysis

[0134] The naive group exhibited the highest variability and the greatest average number of mast cells per hpf in the colon (FIG. 25 A). In contrast, all treatment groups, including Vehicle and both pexacerfont doses, showed lower average mast cell counts compared to the naive group but did not reach significance. In the cecum (FIG. 25B), the71900-429670 vehicle group exhibited the highest variability and average mast cell number, which was significantly increased compared to naive, and tended to be lower with the therapeutic.

Claims

71900-429670WHAT IS CLAIMED IS:

1. A method of reducing formation of one or more adhesions in a patient, the method comprising the step of administering a CRH1 receptor antagonist to the patient, wherein the administration of the CRH1 receptor antagonist reduces formation of adhesions.

2. The method of claim 1, wherein the one or more adhesions comprises an endometriosis-related adhesion.

3. The method of claim 1. wherein the one or more adhesions comprises a pelvic adhesion.

4. The method of claim 1, wherein the one or more adhesions comprises a gastrointestinal adhesion.

5. The method of claim 1, wherein the one or more adhesions comprises a post-surgical adhesion.

6. The method of claim 1, wherein the one or more adhesions are induced by a surgical intervention on the patient.

7. A method of reducing gastrointestinal damage in a patient, the method comprising the step of administering a CRH1 receptor antagonist to the patient, wherein the administration of the CRH1 receptor antagonist reduces gastrointestinal damage.

8. The method of claim 7, wherein the gastrointestinal damage comprises gastrointestinal adhesions.

9. The method of claim 7, wherein the gastrointestinal damage comprises endometriosis-induced gastrointestinal damage.

10. The method of claim 7, wherein the gastrointestinal damage comprises fibrosis.

11. The method of claim 10, wherein the fibrosis comprises colonic fibrosis in the patient.

12. The method of claim 10, wherein the fibrosis comprises cecal fibrosis in the patient.

13. A method of reducing inflammation in a patient, the method comprising the step of administering a CRH1 receptor antagonist to the patient, wherein the administration of the CRH1 receptor antagonist reduces one or more inflammatory signals in the patient.

14. The method of claim 13, wherein the one or more inflammatory signals comprises a decrease in an inflammatory cytokine.

15. The method of claim 14, wherein the inflammatory cytokine is IL-6.

16. The method of claim 14, wherein the inflammatory cytokine is IL-ip.

17. The method of claim 14, wherein the inflammatory cytokine is TNF-a.71900-42967018. The method of claim 13, wherein the one or more inflammatory signals comprises a decrease in mast cells.

19. The method of claim 13, wherein the inflammation comprises colonic inflammation.

20. The method of claim 13, wherein the inflammation comprises cecal inflammation.

21. The method of claim 13, wherein the inflammation comprises fibrosis.

22. The method of claim 21, wherein the fibrosis comprises colonic fibrosis in the patient.

23. The method of claim 21, wherein the fibrosis comprises cecal fibrosis in the patient.

24. A method of reducing pain in a patient, the method comprising the step of administering a CRH1 receptor antagonist to the patient, wherein the administration of the CRH1 receptor antagonist reduces one or more pain symptoms in the patient.

25. The method of claim 24, wherein the reduction in one or more pain symptoms comprises a reduction in inflammatory pain.

26. The method of claim 25, wherein the reduction in inflammatory pain a decrease in an inflammatory cytokine.

27. The method of claim 25, wherein the inflammatory cytokine is IL-6.

28. The method of claim 25, wherein the inflammatory cytokine is IL-ip.

29. The method of claim 25, wherein the inflammatory cytokine is TNF-a.

30. The method of claim 24, wherein the reduction in one or more pain symptoms comprises a reduction in mechanical pain.

31. The method of any one of claims 1 to 30, wherein the patient is male.

32. The method of any one of claims 1 to 30, wherein the patient is female.

33. The method of any one of claims 1 to 30, wherein administering theCRH1 receptor antagonist maintains FSH / LH ratio in the patient.

34. The method of any one of claims 1 to 30, wherein administering theCRH1 receptor antagonist is an acute administration to the patient.

35. The method of any one of claims 1 to 30, wherein administering the CRH1 receptor antagonist is a chronic administration to the patient.

36. The method of any one of claims 1 to 30, wherein administering the CRH1 receptor antagonist is a prophylactic administration to the patient.

37. The method of any one of claims 1 to 30, wherein administering the CRH1 receptor antagonist is an oral administration to the patient.71900-42967038. The method of any one of claims 1 to 30, wherein the CRH1 receptor antagonist is selected from the group consisting of pexacerfont, antalarmin, emicerfont, crinecerfont, tildacerfont, verucerfont (GSK-561679), LWH-234, CP-154,526, NBI-27914, R- 121,919. GW-876008. CRA-1000, NB1-35965. R121919 (NB1-30775), CP-316.

311. R317573 (JNJ- 19567470; CRA-5626), ONO-2333Ms, GSK586529, a-helical CRH (9-41), astressin-2B, and any combination thereof.

39. The method of any one of claims 1 to 30, wherein the CRH1 receptor antagonist is pexacerfont.

Images