Blocking il-33 / st2 signaling to treat endometriosis-associated pain & reduce endometriosis lesions

Abstract

Disclosed herein are compositions comprising an Interleukin-33 (IL-33) inhibitor, an Interleukin-1 receptor-like 1 (ST2) receptor inhibitor or a combination thereof, and methods for treating endometriosis.

Description

[0001] Attorney Docket Number 11624-005W01

[0002] BLOCKING IL-33 / ST2 SIGNALING TO TREAT ENDOMETRIOSIS-ASSOCIATED PAIN & REDUCE ENDOMETRIOSIS LESIONS

[0003] I. CROSS-REFERENCE TO RELATED APPLICATIONS

[0004] 001. This application claims benefit of U.S. Provisional Application No. 63 / 686,349, filed August 23, 2024, incorporated herein by reference in its entirety.

[0005] I. STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH

[0006] 002. This invention was made with government support under Grant No. HT9425-23-1-0040 awarded by the Department of Defense. The Government has certain right in the invention.

[0007] II. BACKGROUND

[0008] 003. Endometriosis is a painful inflammatory disease that affects up to 10% of individuals around the world with annual health care costs approaching $100 billion in the US alone. Because of debilitating chronic pain, affected patients lose around 11 hours of work weekly due to reduced productivity. Current treatments for pain in endometriosis are limited to non-steroidal anti-inflammatory drugs (NSAIDs), other analgesics, hormonal agents, and surgical removal of the lesions. While effective for a fraction of patients, hormonal therapies and NSAIDs present several side effects, and should be used with caution by patients with comorbidities. Moreover, even after surgery, disease and pain recurrence are very common and -15% of individuals experience no relief with medical therapy, and up -30% experience recurrence of pain symptoms after treatment cessation. Therefore, what is needed are new medical therapies and targets thereof, that provide treatment for Endometriosis.

[0009] III. SUMMARY

[0010] 004. The present disclosure relates to compositions used for treatment of endometriosis.

[0011] 005. Disclosed herein are methods of treating, inhibiting, reducing, decreasing, ameliorating and / or preventing endometriosis in a subject, comprising administering to the subject a therapeutically effective amount of a composition comprising an Interleukin-33 (IL-33) inhibitor (such as for example, an anti-IL-33 antibody including, but not limited to, tozorakimab, itepekimab, and SAR440340), a suppressor of tumorigenicity 2 (ST2) receptor inhibitor (such as for example, an anti-ST2 antibody including, but not limited to, astegolimab), or a combination thereof. In one aspect, the composition targets IL-33 or ST2 in macrophages or endometrial epithelial cells.

[0012] 006. Also disclosed herein are methods of treating, inhibiting, reducing, decreasing, Attorney Docket Number 11624-005W01 ameliorating and / or preventing endometriosis of any preceding aspect, wherein administering the composition to the subject, reduces number of endometriosis lesions in a subject, compared to an untreated control.

[0013] 007. In one aspect, disclosed herein are methods of treating, inhibiting, reducing, decreasing, ameliorating and / or preventing endometriosis of any preceding aspect, wherein administering the composition to the subject, reduces number of bouts of abdominal squashing and / or abdominal contortions per 10 minutes, in a subject, compared to an untreated control.

[0014] 008. Also disclosed herein are methods of treating, inhibiting, reducing, decreasing, ameliorating and / or preventing endometriosis lesions in a subject with endometriosis, comprising administering to the subject a therapeutically effective amount of a composition comprising an Interleukin-33 (IL-33) inhibitor (such as for example, an anti-IL-33 antibody including, but not limited to, tozorakimab, itepekimab, and SAR440340), a suppressor of tumorigenicity 2 (ST2) receptor inhibitor (such as for example, an anti-ST2 antibody including, but not limited to, astegolimab), or a combination thereof. In one aspect, the composition targets IL-33 or ST2 in macrophage.

[0015] 009. In one aspect, disclosed herein are methods of treating, inhibiting, reducing, decreasing, ameliorating and / or preventing endometriosis associated nociception in a subject with endometriosis, comprising administering to the subject a therapeutically effective amount of a composition comprising an lnterleukin-33 (IL-33) inhibitor (such as for example, an anti-IL-33 antibody including, but not limited to, tozorakimab, itepekimab, and SAR440340), a suppressor of tumorigenicity 2 (ST2) receptor inhibitor (such as for example, an anti-ST2 antibody including, but not limited to, astegolimab), or a combination thereof. In one aspect, the composition targets IL-33 or ST2 in macrophage.

[0016] IV. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] 010. The accompanying drawings, which are incorporated in and constitute a part of this specification, illustrate several embodiments and together with the description illustrate the disclosed compositions and methods.

[0018] Oil. Figures 1A-1D show that IL-33 / ST2 signaling is required for lesion establishment / formation and endometriosis-associated pain and that blocking 1L-33 / ST-2 signaling is useful in treating endometriosis. Figures 1A, IB, and 1C show the effect of anti-IL- 33 treatment on pain and lesion growth. Figure 1A shows abdominal mechanical hyperalgesia measured using von Frey filaments before (zero) and after (7-28 dpi, weekly) endometriosis Attorney Docket Number 11624-005W01 induction. Results are presented as mean ± SEM of mechanical threshold, n = 10 mice per group (two-way repeated-measures ANOVA followed by Tukey's post hoc, *p<0.05, **p<0.005, ***p< 0.001). Figure IB shows spontaneous pain measured by abdominal squashing and abdominal contortions. Figure 1C shows lesion size, as determined in the remaining lesions by measuring perpendicular diameters. Number of visible lesions was calculated as sum of total lesions per mouse. Lesion burden was calculated as the sum of all lesion size for each mouse. Results are expressed as mean ± SEM, n = 10 (one-way ANOVA followed by Tukey's post hoc, ****p< 0.0001). Figure ID demonstrates the requirement of IL-33 / ST2 signaling for lesion formation. Number of visible lesions was calculated as sum of total lesions per mouse. Lesion size was determined in the remaining lesions by measuring perpendicular diameters. Lesion burden was calculated as the sum of all lesion sizes. Results are expressed as mean ± SEM, n = 15 (WT mice) or 16 (ST2-KO mice) (one-way ANOVA followed by Tukey's post hoc, ****p< 0.0001).

[0019] 012. Figures 2A, 2B, and 2C show the effect of IL-33 treatment (0.002-20 ng / mL) on endometrial epithelial (endo-epi) cell proliferation over 24h, measured using CyQuant as shown in Figure 2A. Figure 2B shows the effect of anti-ST2 antibody treatment (0.001-10 ug / mL) on IL- 33 (2 ng / mL)-induced cell proliferation over 24h, measured using CyQuant. Results are expressed as mean ± SEM, n = 16 (one-way ANOVA followed by Tukey's post hoc or Student's t-test, ****P<0.0001, ***p<0.001, **p<0.01, *p<0.05). Figure 2C shows immunofluorescence images showing ST2 presence in the epithelium of endometriosis lesion at 28 days post implantation (dpi).

[0020] V. DETAILED DESCRIPTION

[0021] 013. Before the present compounds, compositions, articles, devices, and / or methods are disclosed and described, it is to be understood that they are not limited to specific synthetic methods or specific recombinant biotechnology methods unless otherwise specified, or to particular reagents unless otherwise specified, as such may, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting. a. Definitions

[0022] 014. As used in the specification and the appended claims, the singular forms “a,” “an” and “the” include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to “a pharmaceutical carrier” includes mixtures of two or more such carriers, and the like.

[0023] 015. Ranges can be expressed herein as from “about” one particular value, and / or to “about” another particular value. When such a range is expressed, another embodiment includes from Attorney Docket Number 11624-005W01 the one particular value and / or to the other particular value. Similarly, when values are expressed as approximations, by use of the antecedent “about,” it will be understood that the particular value forms another embodiment. It will be further understood that the endpoints of each of the ranges are significant both in relation to the other endpoint, and independently of the other endpoint. It is also understood that there are a number of values disclosed herein, and that each value is also herein disclosed as “about” that particular value in addition to the value itself. For example, if the value “10” is disclosed, then “about 10” is also disclosed. It is also understood that when a value is disclosed that “less than or equal to” the value, “greater than or equal to the value” and possible ranges between values are also disclosed, as appropriately understood by the skilled artisan. For example, if the value “10” is disclosed the “less than or equal to 10”as well as “greater than or equal to 10” is also disclosed. It is also understood that the throughout the application, data is provided in a number of different formats, and that this data, represents endpoints and starting points, and ranges for any combination of the data points. For example, if a particular data point “10” and a particular data point 15 are disclosed, it is understood that greater than, greater than or equal to, less than, less than or equal to, and equal to 10 and 15 are considered disclosed as well as between 10 and 15. It is also understood that each unit between two particular units is also disclosed. For example, if 10 and 15 are disclosed, then 11, 12, 13, and 14 are also disclosed.

[0024] 016. In this specification and in the claims which follow, reference will be made to a number of terms which shall be defined to have the following meanings:

[0025] 017. “Optional” or “optionally” means that the subsequently described event or circumstance may or may not occur, and that the description includes instances where said event or circumstance occurs and instances where it does not.

[0026] 018. “Endometriosis” refers to a condition in which endometrium-like tissue grows in locations outside the uterus. When endometriosis occurs in the ovary, endometriotic cells can form a menstrual fluid-filled sac, which is referred to as “endometrioma.”

[0027] 019. An "increase" can refer to any change that results in a greater amount of a symptom, disease, composition, condition or activity. An increase can be any individual, median, or average increase in a condition, symptom, activity, composition in a statistically significant amount. Thus, the increase can be a 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 100% increase so long as the increase is statistically significant. 020. A "decrease" can refer to any change that results in a smaller amount of a symptom, disease, composition, condition, or activity, for example, a decrease can be a change in the symptoms of a disorder such that the symptoms are less than previously observed. A decrease Attorney Docket Number 11624-005W01 can be any individual, median, or average decrease in a condition, symptom, activity, composition in a statistically significant amount. Thus, the decrease can be a 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 100% decrease so long as the decrease is statistically significant.

[0028] 021. "Inhibit," "inhibiting," and "inhibition" mean to decrease an activity, response, condition, disease, or other biological parameter. This can include but is not limited to the complete ablation of the activity, response, condition, or disease. This may also include, for example, a 10% reduction in the activity, response, condition, or disease as compared to the native or control level. Thus, the reduction can be a 10, 20, 30, 40, 50, 60, 70, 80, 90, 100%, or any amount of reduction in between as compared to native or control levels.

[0029] 022. By “reduce” or other forms of the word, such as “reducing” or “reduction,” is meant lowering of an event or characteristic (e.g., pain, number of endometriosis lesions). It is understood that this is typically in relation to some standard or expected value, in other words it is relative, but that it is not always necessary for the standard or relative value to be referred to. For example, “reduces endometriosis lesions” means reducing the rate of growth and number of endometriosis lesions relative to a standard or a control.

[0030] 023. By “prevent” or other forms of the word, such as “preventing” or “prevention,” is meant to stop a particular event or characteristic, to stabilize or delay the development or progression of a particular event or characteristic, or to minimize the chances that a particular event or characteristic will occur. Prevent does not require comparison to a control as it is typically more absolute than, for example, reduce. As used herein, something could be reduced but not prevented, but something that is reduced could also be prevented. Likewise, something could be prevented but not reduced, but something that is prevented could also be reduced. It is understood that where reduce or prevent are used, unless specifically indicated otherwise, the use of the other word is also expressly disclosed.

[0031] 024. The term “subject” refers to any individual who is the target of administration or treatment. The subject can be a vertebrate, for example, a mammal. In one aspect, the subject can be human, non-human primate, bovine, equine, porcine, canine, or feline. The subject can also be a guinea pig, rat, hamster, rabbit, mouse, or mole. Thus, the subject can be a human or veterinary patient. The term “patient” refers to a subject under the treatment of a clinician, e.g., physician.

[0032] 025. The term “therapeutically effective” refers to the amount of the composition used is of sufficient quantity to ameliorate one or more causes or symptoms of a disease or disorder. Such amelioration only requires a reduction or alteration, not necessarily elimination. Attorney Docket Number 11624-005W01

[0033] 026. The term “treatment” refers to the medical management of a patient with the intent to cure, ameliorate, stabilize, or prevent a disease, pathological condition, or disorder. This term includes active treatment, that is, treatment directed specifically toward the improvement of a disease, pathological condition, or disorder, and also includes causal treatment, that is, treatment directed toward removal of the cause of the associated disease, pathological condition, or disorder. In addition, this term includes palliative treatment, that is, treatment designed for the relief of symptoms rather than the curing of the disease, pathological condition, or disorder; preventative treatment, that is, treatment directed to minimizing or partially or completely inhibiting the development of the associated disease, pathological condition, or disorder; and supportive treatment, that is, treatment employed to supplement another specific therapy directed toward the improvement of the associated disease, pathological condition, or disorder. 027. "Biocompatible" generally refers to a material and any metabolites or degradation products thereof that are generally non-toxic to the recipient and do not cause significant adverse effects to the subject.

[0034] 028. "Comprising" is intended to mean that the compositions, methods, etc. include the recited elements, but do not exclude others. "Consisting essentially of' when used to define compositions and methods, shall mean including the recited elements, but excluding other elements of any essential significance to the combination. Thus, a composition consisting essentially of the elements as defined herein would not exclude trace contaminants from the isolation and purification method and pharmaceutically acceptable carriers, such as phosphate buffered saline, preservatives, and the like. "Consisting of’ shall mean excluding more than trace elements of other ingredients and substantial method steps for administering the compositions provided and / or claimed in this disclosure. Embodiments defined by each of these transition terms are within the scope of this disclosure.

[0035] 029. A “control” is an alternative subject or sample used in an experiment for comparison purposes. A control can be "positive" or "negative."

[0036] 030. “Effective amount” of an agent refers to a sufficient amount of an agent to provide a desired effect. The amount of agent that is “effective” will vary from subject to subject, depending on many factors such as the age and general condition of the subject, the particular agent or agents, and the like. Thus, it is not always possible to specify a quantified “effective amount.” However, an appropriate “effective amount” in any subject case may be determined by one of ordinary skill in the art using routine experimentation. Also, as used herein, and unless specifically stated otherwise, an “effective amount” of an agent can also refer to an amount covering both therapeutically effective amounts and prophylactically effective amounts. Attorney Docket Number 11624-005W01

[0037] An “effective amount” of an agent necessary to achieve a therapeutic effect may vary according to factors such as the age, sex, and weight of the subject. Dosage regimens can be adjusted to provide the optimum therapeutic response. For example, several divided doses may be administered daily, or the dose may be proportionally reduced as indicated by the exigencies of the therapeutic situation.

[0038] 031. A "pharmaceutically acceptable" component can refer to a component that is not biologically or otherwise undesirable, i.e., the component may be incorporated into a pharmaceutical formulation provided by the disclosure and administered to a subject as described herein without causing significant undesirable biological effects or interacting in a deleterious manner with any of the other components of the formulation in which it is contained. When used in reference to administration to a human, the term generally implies the component has met the required standards of toxicological and manufacturing testing or that it is included on the Inactive Ingredient Guide prepared by the U.S. Food and Drug Administration.

[0039] 032. "Pharmaceutically acceptable carrier" (sometimes referred to as a “carrier”) means a carrier or excipient that is useful in preparing a pharmaceutical or therapeutic composition that is generally safe and non-toxic and includes a carrier that is acceptable for veterinary and / or human pharmaceutical or therapeutic use. The terms "carrier" or "pharmaceutically acceptable carrier" can include, but are not limited to, phosphate buffered saline solution, water, emulsions (such as an oil / water or water / oil emulsion) and / or various types of wetting agents. As used herein, the term "carrier" encompasses, but is not limited to, any excipient, diluent, filler, salt, buffer, stabilizer, solubilizer, lipid, stabilizer, or other material well known in the art for use in pharmaceutical formulations and as described further herein.

[0040] 033. “Pharmacologically active” (or simply “active”), as in a “pharmacologically active” derivative or analog, can refer to a derivative or analog (e.g., a salt, ester, amide, conjugate, metabolite, isomer, fragment, etc.) having the same type of pharmacological activity as the parent compound and approximately equivalent in degree.

[0041] 034. “Therapeutic agent” refers to any composition that has a beneficial biological effect. Beneficial biological effects include both therapeutic effects, e.g., treatment of a disorder or other undesirable physiological condition, and prophylactic effects, e.g., prevention of a disorder or other undesirable physiological condition (e.g., a non-immunogenic cancer). The terms also encompass pharmaceutically acceptable, pharmacologically active derivatives of beneficial agents specifically mentioned herein, including, but not limited to, salts, esters, amides, proagents, active metabolites, isomers, fragments, analogs, and the like. When the terms “therapeutic agent” is used, then, or when a particular agent is specifically identified, it is to be Attorney Docket Number 11624-005W01 understood that the term includes the agent per se as well as pharmaceutically acceptable, pharmacologically active salts, esters, amides, proagents, conjugates, active metabolites, isomers, fragments, analogs, etc.

[0042] 035. “Therapeutically effective amount” or “therapeutically effective dose” of a composition (e.g. a composition comprising an agent) refers to an amount that is effective to achieve a desired therapeutic result. In some embodiments, a desired therapeutic result is the decrease in the number of endometriosis lesions. In some embodiments, a desired therapeutic result is the control of pain. Therapeutically effective amounts of a given therapeutic agent will typically vary with respect to factors such as the type and severity of the disorder or disease being treated and the age, gender, and weight of the subject. The term can also refer to an amount of a therapeutic agent, or a rate of delivery of a therapeutic agent (e.g., amount over time), effective to facilitate a desired therapeutic effect, such as pain relief. The precise desired therapeutic effect will vary according to the condition to be treated, the tolerance of the subject, the agent and / or agent formulation to be administered (e.g., the potency of the therapeutic agent, the concentration of agent in the formulation, and the like), and a variety of other factors that are appreciated by those of ordinary skill in the art. In some instances, a desired biological or medical response is achieved following administration of multiple dosages of the composition to the subject over a period of days, weeks, or years.

[0043] 036. Throughout this application, various publications are referenced. The disclosures of these publications in their entireties are hereby incorporated by reference into this application in order to more fully describe the state of the art to which this pertains. The references disclosed are also individually and specifically incorporated by reference herein for the material contained in them that is discussed in the sentence in which the reference is relied upon. b. Compositions

[0044] 037. Disclosed are the components to be used to prepare the disclosed compositions as well as the compositions themselves to be used within the methods disclosed herein. These and other materials are disclosed herein, and it is understood that when combinations, subsets, interactions, groups, etc. of these materials are disclosed that while specific reference of each various individual and collective combinations and permutation of these compounds may not be explicitly disclosed, each is specifically contemplated and described herein. For example, if a particular composition comprising an Interleukin-33 (IL-33) inhibitor, an Interleukin- 1 receptorlike 1 (ST2) receptor inhibitor or a combination thereof is disclosed and discussed and a number of modifications that can be made to a number of combinations including the composition comprising an Interleukin-33 (IL-33) inhibitor, an Interleukin- 1 receptor-like 1 (ST2) receptor Attorney Docket Number 11624-005W01 inhibitor are discussed, specifically contemplated is each and every combination and permutation of composition comprising an Interleukin-33 (IL-33) inhibitor, an Interleukin- 1 receptor-like 1 (ST2) receptor inhibitor and the modifications that are possible unless specifically indicated to the contrary. Thus, if a class of molecules A, B, and C are disclosed as well as a class of molecules D, E, and F and an example of a combination molecule, A-D is disclosed, then even if each is not individually recited each is individually and collectively contemplated meaning combinations, A-E, A-F, B-D, B-E, B-F, C-D, C-E, and C-F are considered disclosed. Likewise, any subset or combination of these is also disclosed. Thus, for example, the sub-group of A-E, B-F, and C-E would be considered disclosed. This concept applies to all aspects of this application including, but not limited to, steps in methods of making and using the disclosed compositions. Thus, if there are a variety of additional steps that can be performed it is understood that each of these additional steps can be performed with any specific embodiment or combination of embodiments of the disclosed methods.

[0045] Method of treating endometriosis, endometriosis lesions, and nociception associated with endometriosis

[0046] 038. Nociceptors are pain-sensing neurons that detect environmental cues and potentially damaging stimuli. However, aberrant activation of nociceptors is associated with the pathogenesis of pain-related disorders such as endometriosis, a painful inflammatory disease that affects up to 10% of individuals globally. Endometriosis is characterized by the development of endometrial-like lesions outside the uterus, which can lead to pain, including spontaneous pain and chronic pelvic pain. Current treatments for endometrial pain are limited to non-steroidal anti-inflammatory drugs (NSAIDs), other analgesics, hormonal agents, and surgical removal of endometrial lesions. While some endometriosis patients experience reduced pain as a result of these treatments, many patients still experience pain, while approximately 15% of patients experience no relief as a result of medical intervention. In one aspect, it is understood and herein contemplated that the methods and compositions disclosed herein can be used to treat any disease where uncontrolled or ectopic cellular proliferation occurs such as endometriosis.

[0047] 039. In one aspect, disclosed herein are methods of treating, inhibiting, reducing, decreasing, ameliorating and / or preventing endometriosis in a subject, comprising administering to the subject a therapeutically effective amount of a composition comprising an Interleukin- 33 (IL-33) inhibitor (such as for example, an anti-IL-33 antibody including, but not limited to, tozorakimab, itepekimab, and SAR440340), a suppressor of tumorigenicity 2 (ST2) receptor inhibitor (such as for example, an anti-ST2 antibody including, but not limited to, astegolimab), or a combination Attorney Docket Number 11624-005W01 thereof. In one aspect, the composition targets IL-33 or ST2 in macrophages or endometrial epithelial cells.

[0048] 040. Also disclosed herein are methods of treating, inhibiting, reducing, decreasing, ameliorating and / or preventing endometriosis, wherein administering the composition to the subject, reduces number of endometriosis lesions in a subject, compared to an untreated control. 041. In one aspect, disclosed herein are methods of treating, inhibiting, reducing, decreasing, ameliorating and / or preventing endometriosis, wherein administering the composition to the subject, reduces number of bouts of abdominal squashing and / or abdominal contortions per 10 minutes, in a subject, compared to an untreated control.

[0049] 042. Also disclosed herein are methods of treating, inhibiting, reducing, decreasing, ameliorating and / or preventing endometriosis lesions in a subject with endometriosis, comprising administering to the subject a therapeutically effective amount of a composition comprising an Interleukin-33 (IL-33) inhibitor (such as for example, an anti-IL-33 antibody including, but not limited to, tozorakimab, itepekimab, and SAR440340), a suppressor of tumorigenicity 2 (ST2) receptor inhibitor (such as for example, an anti-ST2 antibody including, but not limited to, astegolimab), or a combination thereof. In one aspect, the composition targets IL-33 or ST2 in macrophages or endometrial epithelial cells.

[0050] 043. A method of treating, inhibiting, reducing, decreasing, ameliorating and / or preventing endometriosis associated nociception in a subject with endometriosis, comprising administering to the subject a therapeutically effective amount of a composition comprising an Interleukin-33 (IL-33) inhibitor (such as for example, an anti-IL-33 antibody including, but not limited to, tozorakimab, itepekimab, and SAR440340), a suppressor of tumorigenicity 2 (ST2) receptor inhibitor (such as for example, an anti-ST2 antibody including, but not limited to, astegolimab), or a combination thereof. In one aspect, the composition targets IL-33 or ST2 in macrophages or endometrial epithelial cells.

[0051] 044. In some aspects, the IL-33 inhibitor and / or ST2 inhibitor used in the disclosed methods can be any molecule capable of interrupting the interaction and or signaling resulting from IL-33 binding to ST2. Thus, the IL-33 inhibitor and / or ST2 inhibitor can be any small molecule, antibody, nanobody, diabody, antibody fragment (such as for example, scFv), bispecific T cell engager (BiTE), DARPin, peptide, protein, or functional nucleic acid including, but not limited to antisense oligonucleotide, siRNA, shRNA, IncRNA, mRNA, or miRNA that targets IL-33 or ST2.

[0052] 045. In some embodiments, the compositions comprising an Interleukin-33 (IL-33) inhibitor, an Interleukin- 1 receptor- like 1 (ST2) receptor inhibitor or a combination thereof is administered Attorney Docket Number 11624-005W01 to the subject with endometriosis, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19,

[0053] 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45,

[0054] 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71,

[0055] 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,

[0056] 98, 99, 100, or more times. In some embodiments, the compositions comprising an Interleukin- 33 (IL-33) inhibitor, an Interleukin-1 receptor-like 1 (ST2) receptor inhibitor or a combination thereof is administered daily. In some embodiments, the compositions comprising an Interleukin- 33 (IL-33) inhibitor, an Interleukin- 1 receptor- like 1 (ST2) receptor inhibitor or a combination thereof is administered every day, every 2 days, every 3 days, every 4 days, every 5 days, every 6 days, every 7 days, or more. In some embodiments, the compositions comprising an Interleukin-33 (IL-33) inhibitor, an Interleukin- 1 receptor-like 1 (ST2) receptor inhibitor or a combination thereof is administered every week, every 2 weeks, every 3 weeks, every 4 weeks, or more. In some embodiments, the compositions comprising an Interleukin- 33 (IL-33) inhibitor, an Interleukin- 1 receptor-like 1 (ST2) receptor inhibitor or a combination thereof is administered every month, every 2 months, every 3 months, every 4 months, every 5 months, every 6 months, every 7 months, every 8 months, every 9 months, every 10 months, every 11 months, every 12 months, or more. In some embodiments, the compositions comprising an Interleukin-33 (IL-33) inhibitor, an Interleukin- 1 receptor-like 1 (ST2) receptor inhibitor or a combination thereof is administered every year, every 2 years, every 3 years, every 4 years, every 5 years, or more.

[0057] 046. The concentration of active agent(s) can vary widely and are selected primarily based on activity of the active ingredient(s), body weight and the like in accordance with the particular mode of administration selected and the patient's needs. Concentrations, however, can typically be selected to provide dosages ranging from about 0.1 or 1 mg / kg / day to about 50 mg / kg / day and sometimes higher. Typical dosages range from about 3 mg / kg / day to about 3.5 mg / kg / day, preferably from about 3.5 mg / kg / day to about 7.2 mg / kg / day, more preferably from about 7.2 mg / kg / day to about 1 1 .0 mg / kg / day, and most preferably from about 11 .0 mg / kg / day to about 15.0 mg / kg / day. In certain preferred embodiments, dosages range from about 10 mg / kg / day to about 50 mg / kg / day. In certain embodiments, dosages range from about 20 mg to about 50 mg given orally twice daily. It will be appreciated that such dosages may be varied to optimize a therapeutic and / or prophylactic regimen in a particular subject or group of subjects.

[0058] 047. Reducing or inhibiting endometriosis comprises a decrease in the size, proliferation, differentiation, metastasis, and / or activity of the endometriosis lesions in a subject. The decrease can be a 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, Attorney Docket Number 11624-005W01

[0059] 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,

[0060] 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100%, or any amount of reduction in between as compared to a standard or control.

[0061] Antibodies a. Antibodies Generally

[0062] 048. As noted herein, the IL-33 inhibitor and / or ST2 inhibitor can be an antibody or antibody fragment. The term “antibodies” is used herein in a broad sense and includes both polyclonal and monoclonal antibodies. In addition to intact immunoglobulin molecules, also included in the term “antibodies” are fragments or polymers of those immunoglobulin molecules, and human or humanized versions of immunoglobulin molecules or fragments thereof, as long as they are chosen for their ability to interact with IL-33 or ST2 such that IL-33 is inhibited from interacting with ST2. Antibodies that bind the disclosed regions of IL-33 or ST2 involved in the interaction between IL-33 or ST2 are also disclosed. The antibodies can be tested for their desired activity using the in vitro assays described herein, or by analogous methods, after which their in vivo therapeutic and / or prophylactic activities are tested according to known clinical testing methods. There are five major classes of human immunoglobulins: IgA, IgD, IgE, IgG and IgM, and several of these may be further divided into subclasses (isotypes), e.g., IgG-1, IgG-2, IgG-3, and IgG-4; IgA-1 and IgA-2. One skilled in the art would recognize the comparable classes for mouse. The heavy chain constant domains that correspond to the different classes of immunoglobulins are called alpha, delta, epsilon, gamma, and mu, respectively.

[0063] 049. The term “monoclonal antibody” as used herein refers to an antibody obtained from a substantially homogeneous population of antibodies, i.e., the individual antibodies within the population are identical except for possible naturally occurring mutations that may be present in a small subset of the antibody molecules. The monoclonal antibodies herein specifically include "chimeric" antibodies in which a portion of the heavy and / or light chain is identical with or homologous to corresponding sequences in antibodies derived from a particular species or belonging to a particular antibody class or subclass, while the remainder of the chain(s) is identical with or homologous to corresponding sequences in antibodies derived from another species or belonging to another antibody class or subclass, as well as fragments of such antibodies, as long as they exhibit the desired antagonistic activity.

[0064] 050. The disclosed monoclonal antibodies can be made using any procedure which produces monoclonal antibodies. For example, disclosed monoclonal antibodies can be prepared using hybridoma methods, such as those described by Kohler and Milstein, Nature, 256:495 (1975). Attorney Docket Number 11624-005W01

[0065] In a hybridoma method, a mouse or other appropriate host animal is typically immunized with an immunizing agent to elicit lymphocytes that produce or are capable of producing antibodies that will specifically bind to the immunizing agent. Alternatively, the lymphocytes may be immunized in vitro.

[0066] 051. The monoclonal antibodies may also be made by recombinant DNA methods. DNA encoding the disclosed monoclonal antibodies can be readily isolated and sequenced using conventional procedures (e.g., by using oligonucleotide probes that are capable of binding specifically to genes encoding the heavy and light chains of murine antibodies). Libraries of antibodies or active antibody fragments can also be generated and screened using phage display techniques, e.g., as described in U.S. Patent No. 5,804,440 to Burton et al. and U.S. Patent No. 6,096,441 to Barbas et al.

[0067] 052. In vitro methods are also suitable for preparing monovalent antibodies. Digestion of antibodies to produce fragments thereof, particularly Fab fragments, can be accomplished using routine techniques known in the art. For instance, digestion can be performed using papain. Examples of papain digestion are described in WO 94 / 29348 published Dec. 22, 1994, and U.S. Pat. No. 4,342,566. Papain digestion of antibodies typically produces two identical antigen binding fragments, called Fab fragments, each with a single antigen binding site, and a residual Fc fragment. Pepsin treatment yields a fragment that has two antigen combining sites and is still capable of cross-linking antigen.

[0068] 053. As used herein, the term “antibody or fragments thereof’ encompasses chimeric antibodies and hybrid antibodies, with dual or multiple antigen or epitope specificities, and fragments, such as F(ab’)2, Fab’, Fab, Fv, sFv, scFv, and the like, including hybrid fragments. Thus, fragments of the antibodies that retain the ability to bind their specific antigens are provided. For example, fragments of antibodies which maintain IL-33 or ST2 binding activity are included within the meaning of the term “antibody or fragment thereof.” Such antibodies and fragments can be made by techniques known in the art and can be screened for specificity and activity according to the methods set forth in the Examples and in general methods for producing antibodies and screening antibodies for specificity and activity (See Harlow and Lane. Antibodies, A Laboratory Manual. Cold Spring Harbor Publications, New York, (1988)).

[0069] 054. Also included within the meaning of “antibody or fragments thereof’ are conjugates of antibody fragments and antigen binding proteins (single chain antibodies).

[0070] 055. The fragments, whether attached to other sequences or not, can also include insertions, deletions, substitutions, or other selected modifications of particular regions or specific amino acids residues, provided the activity of the antibody or antibody fragment is not significantly Attorney Docket Number 11624-005W01 altered or impaired compared to the non-modified antibody or antibody fragment. These modifications can provide for some additional property, such as to remove / add amino acids capable of disulfide bonding, to increase its bio-longevity, to alter its secretory characteristics, etc. In any case, the antibody or antibody fragment must possess a bioactive property, such as specific binding to its cognate antigen. Functional or active regions of the antibody or antibody fragment may be identified by mutagenesis of a specific region of the protein, followed by expression and testing of the expressed polypeptide. Such methods are readily apparent to a skilled practitioner in the art and can include site-specific mutagenesis of the nucleic acid encoding the antibody or antibody fragment. (Zoller, M.J. Curr. Opin. Biotechnol. 3:348-354, 1992).

[0071] 056. As used herein, the term “antibody” or “antibodies” can also refer to a human antibody and / or a humanized antibody. Many non-human antibodies (e.g., those derived from mice, rats, or rabbits) are naturally antigenic in humans, and thus can give rise to undesirable immune responses when administered to humans. Therefore, the use of human or humanized antibodies in the methods serves to lessen the chance that an antibody administered to a human will evoke an undesirable immune response. b. Human antibodies

[0072] 057. The disclosed human antibodies can be prepared using any technique. The disclosed human antibodies can also be obtained from transgenic animals. For example, transgenic, mutant mice that are capable of producing a full repertoire of human antibodies, in response to immunization, have been described (see, e.g., Jakobovits et al., Proc. Natl. Acad. Sci. USA, 90:2551-255 (1993); Jakobovits et al., Nature, 362:255-258 (1993); Bruggermann et al., Year in Immunol., 7:33 (1993)). Specifically, the homozygous deletion of the antibody heavy chain joining region (J(H)) gene in these chimeric and germ-line mutant mice results in complete inhibition of endogenous antibody production, and the successful transfer of the human germ-line antibody gene array into such germ-line mutant mice results in the production of human antibodies upon antigen challenge. Antibodies having the desired activity are selected using Env-CD4-co-receptor complexes as described herein. c. Humanized antibodies

[0073] 058. Antibody humanization techniques generally involve the use of recombinant DNA technology to manipulate the DNA sequence encoding one or more polypeptide chains of an antibody molecule. Accordingly, a humanized form of a non-human antibody (or a fragment thereof) is a chimeric antibody or antibody chain (or a fragment thereof, such as an sFv, Fv, Fab, Attorney Docket Number 11624-005W01

[0074] Fab’, F(ab’)2, or other antigen-binding portion of an antibody) which contains a portion of an antigen binding site from a non-human (donor) antibody integrated into the framework of a human (recipient) antibody.

[0075] 059. To generate a humanized antibody, residues from one or more complementarity determining regions (CDRs) of a recipient (human) antibody molecule are replaced by residues from one or more CDRs of a donor (non-human) antibody molecule that is known to have desired antigen binding characteristics (e.g., a certain level of specificity and affinity for the target antigen). In some instances, Fv framework (FR) residues of the human antibody are replaced by corresponding non-human residues. Humanized antibodies may also contain residues which are found neither in the recipient antibody nor in the imported CDR or framework sequences. Generally, a humanized antibody has one or more amino acid residues introduced into it from a source which is non-human. In practice, humanized antibodies are typically human antibodies in which some CDR residues and possibly some FR residues are substituted by residues from analogous sites in rodent antibodies. Humanized antibodies generally contain at least a portion of an antibody constant region (Fc), typically that of a human antibody (Jones et al., Nature, 321:522-525 (1986), Reichmann et al., Nature, 332:323-327 (1988), and Presta, Curr. Opin. Struct. Biol., 2:593-596 (1992)).

[0076] 060. Methods for humanizing non-human antibodies are well known in the art. For example, humanized antibodies can be generated according to the methods of Winter and co-workers (Jones et al., Nature, 321:522-525 (1986), Riechmann et al., Nature, 332:323-327 (1988), Verhoeyen et al., Science, 239: 1534-1536 (1988)), by substituting rodent CDRs or CDR sequences for the corresponding sequences of a human antibody. Methods that can be used to produce humanized antibodies are also described in U.S. Patent No. 4,816,567 (Cabilly et al.), U.S. Patent No. 5,565,332 (Hoogenboom et al.), U.S. Patent No. 5,721,367 (Kay et al.), U.S. Patent No. 5,837,243 (Deo et al.), U.S. Patent No. 5, 939,598 (Kucherlapati et al.), U.S. Patent No. 6,130,364 (Jakobovits et al.), and U.S. Patent No. 6,180,377 (Morgan et al.). d. Administration of antibodies

[0077] 061. Administration of the antibodies can be done as disclosed herein. Nucleic acid approaches for antibody delivery also exist. The broadly neutralizing anti IL-33 or ST2 antibodies and antibody fragments can also be administered to patients or subjects as a nucleic acid preparation (e.g., DNA or RNA) that encodes the antibody or antibody fragment, such that the patient's or subject's own cells take up the nucleic acid and produce and secrete the encoded antibody or antibody fragment. The delivery of the nucleic acid can be by any means, as disclosed herein, for example. Attorney Docket Number 11624-005W01

[0078] Functional Nucleic Acids

[0079] 062. As noted herein, the IL-33 inhibitor and / or ST2 inhibitor can be a functional nucleic acid (including, but not limited to antisense oligonucleotide, siRNA, shRNA, IncRNA, mRNA, or miRNA). Functional nucleic acids are nucleic acid molecules that have a specific function, such as binding a target molecule or catalyzing a specific reaction. Functional nucleic acid molecules can be divided into the following categories, which are not meant to be limiting. For example, functional nucleic acids include antisense molecules, aptamers, ribozymes, triplex forming molecules, and external guide sequences. The functional nucleic acid molecules can act as affectors, inhibitors, modulators, and stimulators of a specific activity possessed by a target molecule, or the functional nucleic acid molecules can possess a de novo activity independent of any other molecules.

[0080] 063. Functional nucleic acid molecules can interact with any macromolecule, such as DNA, RNA, polypeptides, or carbohydrate chains. Thus, functional nucleic acids can interact with the mRNA of any of the disclosed nucleic acids, such as IL-33 or ST2, and the nucleic acids used for the generation of IL-33 or ST2 knockouts, or the genomic DNA of any of the disclosed nucleic acids, such as IL-33 or ST2, and the nucleic acids used for the generation of IL-33 or ST2 knockouts or they can interact with the polypeptide encoded by any of the disclosed nucleic acids, such as IL-33 or ST2, and the nucleic acids used for the generation of IL-33 or ST2 knockouts. Often functional nucleic acids are designed to interact with other nucleic acids based on sequence homology between the target molecule and the functional nucleic acid molecule. In other situations, the specific recognition between the functional nucleic acid molecule and the target molecule is not based on sequence homology between the functional nucleic acid molecule and the target molecule, but rather is based on the formation of tertiary structure that allows specific recognition to take place.

[0081] 064. Antisense molecules are designed to interact with a target nucleic acid molecule through either canonical or non-canonical base pairing. The interaction of the antisense molecule and the target molecule is designed to promote the destruction of the target molecule through, for example, RNAseH mediated RNA-DNA hybrid degradation. Alternatively, the antisense molecule is designed to interrupt a processing function that normally would take place on the target molecule, such as transcription or replication. Antisense molecules can be designed based on the sequence of the target molecule. Numerous methods for optimization of antisense efficiency by finding the most accessible regions of the target molecule exist. Exemplary methods would be in vitro selection experiments and DNA modification studies using DMS and DEPC. It is preferred that antisense molecules bind the target molecule with a dissociation Attorney Docket Number 11624-005W01 constant (kd)less than or equal to 10‘6, 10’8, IO10, or 10‘12. A representative sample of methods and techniques which aid in the design and use of antisense molecules can be found in the following non-limiting list of United States patents: 5,135,917, 5,294,533, 5,627,158, 5,641,754, 5,691,317, 5,780,607, 5,786,138, 5,849,903, 5,856,103, 5,919,772, 5,955,590, 5,990,088, 5,994,320, 5,998,602, 6,005,095, 6,007,995, 6,013,522, 6,017,898, 6,018,042, 6,025,198, 6,033,910, 6,040,296, 6,046,004, 6,046,319, and 6,057,437.

[0082] Pharmaceutical carriers / Delivery of pharmaceutical products

[0083] 065. As described above, the compositions can also be administered in vivo in a pharmaceutically acceptable carrier. By "pharmaceutically acceptable" is meant a material that is not biologically or otherwise undesirable, i.e., the material may be administered to a subject, along with the nucleic acid or vector, without causing any undesirable biological effects or interacting in a deleterious manner with any of the other components of the pharmaceutical composition in which it is contained. The carrier would naturally be selected to minimize any degradation of the active ingredient and to minimize any adverse side effects in the subject, as would be well known to one of skill in the art.

[0084] 066. The compositions may be administered orally, parenterally (e.g., intravenously), by intramuscular injection, by intraperitoneal injection, transdermally, extracorporeally, topically or the like, including topical intranasal administration or administration by inhalant. As used herein, "topical intranasal administration" means delivery of the compositions into the nose and nasal passages through one or both of the nares and can comprise delivery by a spraying mechanism or droplet mechanism, or through aerosolization of the nucleic acid or vector. Administration of the compositions by inhalant can be through the nose or mouth via delivery by a spraying or droplet mechanism. Delivery can also be directly to any area of the respiratory system (e.g., lungs) via intubation. The exact amount of the compositions required will vary from subject to subject, depending on the species, age, weight and general condition of the subject, the severity of the allergic disorder being treated, the particular nucleic acid or vector used, its mode of administration and the like. Thus, it is not possible to specify an exact amount for every composition. However, an appropriate amount can be determined by one of ordinary skill in the art using only routine experimentation given the teachings herein.

[0085] 067. Parenteral administration of the composition, if used, is generally characterized by injection. Injectables can be prepared in conventional forms, either as liquid solutions or suspensions, solid forms suitable for solution of suspension in liquid prior to injection, or as emulsions. A more recently revised approach for parenteral administration involves use of a Attorney Docket Number 11624-005W01 slow release or sustained release system such that a constant dosage is maintained. See, e.g., U.S. Patent No. 3,610,795, which is incorporated by reference herein.

[0086] 068. The materials may be in solution, suspension (for example, incorporated into microparticles, liposomes, or cells). These may be targeted to a particular cell type via antibodies, receptors, or receptor ligands. The following references are examples of the use of this technology to target specific proteins to tumor tissue (Senter, et al., Bioconjugate Chem., 2:447-451, (1991); Bagshawe, K.D., Br. J. Cancer, 60:275-281, (1989); Bagshawe, et al., Br. J. Cancer, 58:700-703, (1988); Senter, et al., Bioconjugate Chem., 4:3-9, (1993); Battelli, et al., Cancer Immunol. Immunother., 35:421-425, (1992); Pietersz and McKenzie, Immunolog. Reviews, 129:57-80, (1992); and Roffler, et al., Biochem. Pharmacol, 42:2062-2065, (1991)). Vehicles such as "stealth" and other antibody conjugated liposomes (including lipid mediated drug targeting to colonic carcinoma), receptor mediated targeting of DNA through cell specific ligands, lymphocyte directed tumor targeting, and highly specific therapeutic retroviral targeting of murine glioma cells in vivo. The following references are examples of the use of this technology to target specific proteins to tumor tissue (Hughes et al., Cancer Research, 49:6214- 6220, (1989); and Litzinger and Huang, Biochimica et Biophysica Acta, 1104: 179-187, (1992)). In general, receptors are involved in pathways of endocytosis, either constitutive or ligand induced. These receptors cluster in clathrin-coated pits, enter the cell via clathrin-coated vesicles, pass through an acidified endosome in which the receptors are sorted, and then either recycle to the cell surface, become stored intracellularly, or are degraded in lysosomes. The internalization pathways serve a variety of functions, such as nutrient uptake, removal of activated proteins, clearance of macromolecules, opportunistic entry of viruses and toxins, dissociation and degradation of ligand, and receptor- level regulation. Many receptors follow more than one intracellular pathway, depending on the cell type, receptor concentration, type of ligand, ligand valency, and ligand concentration. Molecular and cellular mechanisms of receptor-mediated endocytosis have been reviewed (Brown and Greene, DNA and Cell Biology 10:6, 399-409 (1991)). a) Pharmaceutically Acceptable Carriers

[0087] 069. The compositions, including antibodies, can be used therapeutically in combination with a pharmaceutically acceptable carrier.

[0088] 070. Suitable carriers and their formulations are described in Remington: The Science and Practice of Pharmacy (19th ed.) ed. A.R. Gennaro, Mack Publishing Company, Easton, PA 1995. Typically, an appropriate amount of a pharmaceutically-acceptable salt is used in the formulation to render the formulation isotonic. Examples of the pharmaceutically-acceptable Attorney Docket Number 11624-005W01 carrier include, but are not limited to, saline, Ringer's solution and dextrose solution. The pH of the solution is preferably from about 5 to about 8, and more preferably from about 7 to about 7.5. Further carriers include sustained release preparations such as semipermeable matrices of solid hydrophobic polymers containing the antibody, which matrices are in the form of shaped articles, e.g., films, liposomes or microparticles. It will be apparent to those persons skilled in the art that certain carriers may be more preferable depending upon, for instance, the route of administration and concentration of composition being administered.

[0089] 071. Pharmaceutical carriers are known to those skilled in the art. These most typically would be standard carriers for administration of drugs to humans, including solutions such as sterile water, saline, and buffered solutions at physiological pH. The compositions can be administered intramuscularly or subcutaneously. Other compounds will be administered according to standard procedures used by those skilled in the art.

[0090] 072. Pharmaceutical compositions may include carriers, thickeners, diluents, buffers, preservatives, surface active agents and the like in addition to the molecule of choice. Pharmaceutical compositions may also include one or more active ingredients such as antimicrobial agents, antiinflammatory agents, anesthetics, and the like.

[0091] 073. The pharmaceutical composition may be administered in a number of ways depending on whether local or systemic treatment is desired, and on the area to be treated. Administration may be topically (including ophthalmically, vaginally, rectally, intranasally), orally, by inhalation, or parenterally, for example by intravenous drip, subcutaneous, intraperitoneal or intramuscular injection. The disclosed antibodies can be administered intravenously, intraperitoneally, intramuscularly, subcutaneously, intracavity, or transdermally.

[0092] 074. Preparations for parenteral administration include sterile aqueous or non-aqueous solutions, suspensions, and emulsions. Examples of non-aqueous solvents are propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate. Aqueous carriers include water, alcoholic / aqueous solutions, emulsions or suspensions, including saline and buffered media. Parenteral vehicles include sodium chloride solution, Ringer's dextrose, dextrose and sodium chloride, lactated Ringer's, or fixed oils. Intravenous vehicles include fluid and nutrient replenishers, electrolyte replenishers (such as those based on Ringer’s dextrose), and the like. Preservatives and other additives may also be present such as, for example, antimicrobials, anti-oxidants, chelating agents, inert gases and the like.

[0093] 075. Formulations for topical administration may include ointments, lotions, creams, gels, drops, suppositories, sprays, liquids and powders. Conventional pharmaceutical carriers, aqueous, powder or oily bases, thickeners and the like may be necessary or desirable. Attorney Docket Number 11624-005W01

[0094] 076. Compositions for oral administration include powders or granules, suspensions or solutions in water or non-aqueous media, capsules, sachets, or tablets. Thickeners, flavorings, diluents, emulsifiers, dispersing aids or binders may be desirable..

[0095] 077. Some of the compositions may potentially be administered as a pharmaceutically acceptable acid- or base- addition salt, formed by reaction with inorganic acids such as hydrochloric acid, hydrobromic acid, perchloric acid, nitric acid, thiocyanic acid, sulfuric acid, and phosphoric acid, and organic acids such as formic acid, acetic acid, propionic acid, glycolic acid, lactic acid, pyruvic acid, oxalic acid, malonic acid, succinic acid, maleic acid, and fumaric acid, or by reaction with an inorganic base such as sodium hydroxide, ammonium hydroxide, potassium hydroxide, and organic bases such as mono-, di-, trialkyl and aryl amines and substituted ethanolamines.

[0096] 2. Therapeutic Uses

[0097] 078. Effective dosages and schedules for administering the compositions may be determined empirically, and making such determinations is within the skill in the art. The dosage ranges for the administration of the compositions are those large enough to produce the desired effect in which the symptoms of the disorder are effected. The dosage should not be so large as to cause adverse side effects, such as unwanted cross-reactions, anaphylactic reactions, and the like. Generally, the dosage will vary with the age, condition, sex and extent of the disease in the patient, route of administration, or whether other drugs are included in the regimen, and can be determined by one of skill in the art. The dosage can be adjusted by the individual physician in the event of any counterindications. Dosage can vary, and can be administered in one or more dose administrations daily, for one or several days. Guidance can be found in the literature for appropriate dosages for given classes of pharmaceutical products. For example, guidance in selecting appropriate doses for antibodies can be found in the literature on therapeutic uses of antibodies, e.g., Handbook of Monoclonal Antibodies, Ferrone et al., eds., Noges Publications, Park Ridge, N.J., (1985) ch. 22 and pp. 303-357; Smith et al., Antibodies in Human Diagnosis and Therapy, Haber et al., eds., Raven Press, New York (1977) pp. 365-389. A typical daily dosage of the antibody used alone might range from about 1 pg / kg to up to 100 mg / kg of body weight or more per day, depending on the factors mentioned above.

[0098] VI. EXAMPLES

[0099] 079. The following examples are put forth so as to provide those of ordinary skill in the art with a complete disclosure and description of how the compounds, compositions, articles, devices and / or methods claimed herein are made and evaluated and are intended to be purely Attorney Docket Number 11624-005W01 exemplary and are not intended to limit the disclosure. Efforts have been made to ensure accuracy with respect to numbers (e.g., amounts, temperature, etc.), but some errors and deviations should be accounted for. Unless indicated otherwise, parts are parts by weight, temperature is in °C or is at ambient temperature, and pressure is at or near atmospheric.

[0100] 1. Example 1

[0101] 080. IL-33 is a pleiotropic cytokine with varied immune functions. IL-33 signaling occurs through the interaction with its receptor ST2. The receptor complex for IL-33 consists of the specific subunit of ST2, encoded by the IL1RL1, and the coreceptor, IL-1 receptor accessory protein (IL-lRAcP). Dysregulation of this pathway has been described as a key step in varied immune responses. Further, IL-33 contributes to peripheral and spinal cord nociceptor neuron sensitization in innate and adaptive inflammatory immune responses as well as in neuropathic and cancer pain. In this sense, targeting IL-33 / ST2 signaling is a promising therapeutic approach. This study indicates that, upon stimulus with a neuropeptide called calcitonin gene- related peptide (CGRP), macrophages release IL-33. It was then hypothesized that IL-33 production by macrophages drives endometriosis pain and lesion formation.

[0102] 081. To test this hypothesis, first endometriosis was induced, and an anti-IL-33 antibody was used in a pre-treatment regimen. Endometriosis lesions were induced in 30 mice and 2h before induction, mice were randomly assigned to treatment with IgG control or anti-IL-33 at 0.25 or 2.5 pg / kg (Figure 1 A, n = 10 mice per group). Treatment was performed every other day (s.c.). Mechanical abdominal pain was determined weekly, 7 to 28 dpi using von Frey filaments (Figure 1 A, analyzed by two-way repeated-measures ANOVA followed by Tukey's post hoc) while abdominal squashing and contortions were determined by counting these pain behaviors (Figure IB, analyzed by one-way repeated-measures ANOVA followed by Tukey's post hoc). Lesion size was calculated from the mean of two measurements (width and height in mm) and lesion burden as the total sum of the lesion size (n = 20 mice per group). These methods were in accordance with previous findings. It was found that treatment with anti-IL-33 at 2.5 pg / mouse reduced mechanical pain (Figure 1A) as observed by increase in the threshold. It also reduced spontaneous pain, as observed by reduction in abdominal contortions and squashing (Figure IB). It was also found that treated mice had reduced number of lesions and that these lesions were also smaller, resulting in a reduced lesion burden (Figure 1C). To expand these results and further determine that IL-33 / ST2 signaling is required for lesion formation, endometriosis was induced using uterine tissue from mice lacking ST2 (IL-33 receptor subunit). For that, endometriosis was induced in Balb / c mice using WT tissue (n = 15) or ST2-KO tissue (n = 16) as donors. When endometriosis was induced with uterine tissue from ST2-KO mice, it was Attorney Docket Number 11624-005W01 found that there were reduced number of lesions as well as smaller lesions (D). Collectively, the data show that IL-33 / ST2 signaling is required for lesion stablishment / formation and endometriosis-associated pain and that blocking IL-33 / ST-2 signaling is useful in treating endometriosis.

[0103] 082. To expand these results and further determine that IL-33 / ST2 signaling is required for lesion formation, endometriosis was then induced using uterine tissue from mice lacking ST2 (IL-33 receptor subunit). For that, endometriosis was induced in Balb / c mice using WT tissue (n = 15) or ST2-KO tissue (n - 16) as donors. When endometriosis was induced with uterine tissue from ST2-KO mice, reduced number of lesions as well as smaller lesions were found (Figure ID). Collectively, these data show that IL-33 / ST2 signaling is required for lesion establishment / formation and endometriosis-associated pain and that blocking IL-33 / ST-2 signaling is useful in treating endometriosis.

[0104] 083. To confirm that IL-33 induces cell proliferation, a proliferation assay was performed using mouse endometrial epithelial cells as shown in Figure 2 A. It was also found that IL-33 induces cell proliferation in a concentration-dependent manner and that blocking this communication with anti-ST2 (antibody against IL-33 receptor) reduced cell proliferation (right).

[0105] 084. To confirm that in vivo, endometriosis lesions were induced and immunofluorescence for ST2 and Ki67 (a marker of proliferating cells) was performed as shown in Figure 2C. It was found that found that ST2 is mainly present in the epithelium of the lesions, and that ST2 colocalizes with Ki67, indicating that these ST2+ cells are proliferating.

[0106] 085. In conclusion, it was demonstrated that blocking IL-33 / ST2 signaling using an anti-IL-33 antibody reduces pain and lesion size. Moreover, it was found that IL-33 induces cell proliferation in an ST2-dependent manner as treatment with soluble ST2 reduces endo-epi cell proliferation as shown in Figure 2B. Based on these data, it is shown that other IL-33- or ST2- targeting drugs are also effective at reducing endometriosis burden. Drugs such as tozorakimab, itepekimab, and SAR440340 (anti-IL-33) as well as astegolimab (anti-ST2) are currently under phase 3 clinical trials for the treatment of different diseases such asthma and chronic obstructive pulmonary disease (COPD). Based on these data with anti-IL-33 and ST2 antibodies, it is shown that these drugs are effective at treating endometriosis pain. Therefore, IL-33 to ST2 communication is a non-hormonal and non-opioid approach for endometriosis treatment.

Claims

Attorney Docket Number 11624-005W01VII. CLAIMSWhat is claimed is:

1. A method of treating endometriosis in a subject, comprising administering to the subject a therapeutically effective amount of a composition comprising an Interleukin-33 (IL-33) inhibitor, a suppressor of tumorigenicity 2 (ST2) receptor inhibitor, or a combination thereof.

2. The method of claim 1, wherein the IL-33 inhibitor is an anti-IL-33 antibody.

3. The method of claim 2, wherein the anti-IL-33 antibody is tozorakimab, itepekimab, or SAR440340.

4. The method of any one of claims 1-3, wherein the ST2 receptor inhibitor is an anti-ST2 antibody.

5. The method of claim 4, wherein the anti-ST2 antibody is astegolimab.

6. The method of any one of claims 1-5, wherein the composition targets IL-33 or ST2 in macrophage.

7. The method of any one of claims 1-6, wherein administering the composition to the subject, reduces number of endometriosis lesions in a subject, compared to an untreated control.

8. The method of any one of claims 1-7, wherein administering the composition to the subject, reduces number of bouts of abdominal squashing per 10 minutes, in a subject, compared to an untreated control.

9. The method of any one of claims 1-8, wherein administering the composition to the subject, reduces number of bouts of abdominal contortions per 10 minutes, in a subject, compared to an untreated control.

10. The method of any one of claims 1-9, wherein the subject is a human.

11. A method of reducing endometriosis lesions in a subject with endometriosis, comprising administering to the subject a therapeutically effective amount of a compositionAttorney Docket Number 11624-005W01 comprising an Interleukin-33 (IL-33) inhibitor, a suppressor of tumorigenicity 2 (ST2) receptor inhibitor, or a combination thereof.

12. The method of claim 11, wherein the IL-33 inhibitor is an anti-IL-33 antibody.

13. The method of claim 12, wherein the anti-IL-33 antibody is tozorakimab, itepekimab, or SAR440340.

14. The method of any one of claims 11-13, wherein the ST2 receptor inhibitor is an anti- ST2 antibody.

15. The method of claim 14, wherein the anti-ST2 antibody is astegolimab.

16. The method of any one of claims 11-15, wherein the composition targets IL-33 or ST2 in macrophage.

17. The method of any one of claims 11-16, wherein the subject is a human.

18. A method of reducing endometriosis associated nociception in a subject with endometriosis, comprising administering to the subject a therapeutically effective amount of a composition comprising an Interleukin-33 (IL-33) inhibitor, a suppressor of tumorigenicity 2 (ST2) receptor inhibitor, or a combination thereof.

19. The method of claim 18, wherein the IL-33 inhibitor is an anti-IL-33 antibody.

20. The method of claim 19, wherein the anti-IL-33 antibody is tozorakimab, itepekimab, or SAR440340.

21. The method of any one of claims 18-20, wherein the ST2 receptor inhibitor is an anti- ST2 antibody.

22. The method of claim 21, wherein the anti-ST2 antibody is astegolimab.

23. The method of any one of claims 18-22, wherein the composition targets IL-33 or ST2 in macrophage.

24. The method of any one of claims 18-23, wherein the subject is a human.

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