Methods and medicaments for treating endometriosis

Abstract

Provided are methods and medicaments for treating endometriosis in an individual in need thereof, the methods comprising administering to the individual an effective amount of a hematopoietic cell kinase inhibitor.

Description

[0001] Cross-reference to related applications

[0002] This application claims priority to PCT International Application No. PCT / CN2023 / 112275, filed on August 10, 2023, which is incorporated herein by reference in its entirety.

[0003] field

[0004] This invention relates to the medical field, and in particular to a method for treating endometriosis.

[0005] background

[0006] Endometriosis is a chronic inflammatory gynecological disease characterized by the presence of ectopic endometrial glands and stroma, primarily located in the pelvic peritoneum, ovaries, and rectovaginal septum. It is estimated that approximately 5-15% of women of reproductive age worldwide have endometriosis (PMID: 27159755), while the prevalence can reach 35-80% in women experiencing pelvic pain and / or infertility (PMID: 32212520, 33640070). Due to the variable symptoms and the ease with which endometriosis symptoms can be confused with those of other diseases, diagnosis is often delayed, with an average delay of 4 to 12 years (PMID: 8671344, 22990516, 12790847, 28440744). The etiology of endometriosis is unclear, and several hypotheses have been proposed to explain the origin of this disease (PMID: 30026507 review). From a pathological perspective, genetic factors, environmental factors, immune dysfunction, and estrogen imbalances can all contribute to the formation and maintenance of endometriosis lesions (PMID:30026507). Among these, steroid production disorder is the most studied mechanism, which can manifest as abnormal changes in hormone levels, and abnormal hormone levels are a common risk factor for this disease.

[0007] Due to the complexity and diversity of its pathogenesis, endometriosis remains incurable. Various treatment methods have been employed to address both endometriosis and infertility, including surgery, medication, and acupuncture. Hormone therapy is considered the first-line treatment for endometriosis. In multiple Phase III clinical trials, two approved hormonal drugs, Elagolix and Myfembree, demonstrated their effectiveness in relieving pain caused by endometriosis without any safety issues (PMID: 28525302, 29889764, 34134684, 33066973, 35717987).

[0008] Although endometriosis is not fatal, chronic pain and infertility can significantly impact quality of life. It is estimated that the economic burden of endometriosis in the United States alone reaches $80 billion annually (PMID: 35620300). Furthermore, mounting evidence suggests that endometriosis is a systemic disease that disrupts cardiovascular, neurological, metabolic, and immune function (PMID: 33640070). Patients may have an increased risk of developing a variety of chronic diseases, such as adenomyosis (PMID: 24532217), ovarian cancer (PMID: 28240000), and autoimmune diseases (PMID: 31260048). These factors collectively underscore the need for innovative and effective treatments for this disease. To address this challenge, we utilized the AI-driven target discovery platform PandaOmics to identify novel drug targets for the treatment of endometriosis. Figure 1A As shown, PandaOmics identified hematopoietic kinase as the number one potential therapeutic target using 11 endometriosis transcriptome datasets. The upregulation of hematopoietic kinase expression in human endometriosis samples was further confirmed at both the mRNA and protein levels, and validated through in vitro and in vivo experiments. This study is the first to demonstrate the application of AI in exploring therapeutic targets for endometriosis.

[0009] Overview

[0010] The purpose of this invention is to provide a treatment or medicine for endometriosis.

[0011] One aspect of the present invention provides a method for treating endometriosis in an individual in need, comprising administering an effective amount of a hematopoietic cell kinase inhibitor to the individual.

[0012] In some implementations, the hematopoietic cell kinase inhibitor is selected from one or more of bosutinib, dasatinib, KIN-8194, RV568, rebastinib, and their derivatives.

[0013] Another aspect of the invention provides the use of hematopoietic cell kinase inhibitors in the preparation of medicaments for the treatment of endometriosis.

[0014] In some embodiments, the hematopoietic cell kinase inhibitor is one or more selected from bosutinib, dasatinib, KIN-8194, RV568, rabastinibril and their derivatives.

[0015] Another aspect of the invention provides a medicament for treating endometriosis, wherein the medicament comprises a hematopoietic cell kinase inhibitor.

[0016] In some embodiments, the S1PR1 antagonist is one or more selected from bosutinib, dasatinib, KIN-8194, RV568, rabastinibril and their derivatives. Brief description of the attached diagram

[0018] Figure 1A-1B PandaOmics identified hematopoietic cell kinases as a potential therapeutic target for endometriosis. Figure 1A This document describes the workflow for identifying therapeutic targets for endometriosis using PandaOmics. By inputting 11 endometriosis datasets, PandaOmics prioritized endometriosis targets using a combination of Omics and text scoring (target identification). The platform also performed differential gene expression (DEG) and pathway analysis. After target identification, candidate targets were validated using immunohistochemistry (IHC). Only validated targets proceeded to subsequent in vitro and in vivo siRNA validation stages. Downstream effectors of these validated targets were then predicted using computational and experimental methods. Figure 1B This is a screenshot of the target identification page from the PandaOmics platform's meta-analysis of endometriosis. The results show that hematopoietic cell kinases are novel druggable targets for endometriosis. This analysis used gene expression, genetics, drug, and text data.

[0019] Figure 2A-2C Persistent dysregulation of hematopoietic cell kinases in endometriosis. Figure 2A The expression of hematopoietic cell kinases in 11 comparisons associated with endometriosis is presented in box plot form. Figure 2B Representative paraffin-embedded sections of human endometriotic tissue and normal endometrial stroma were stained with an anti-hematopoietic cell kinase antibody. Figure 2C Protein expression of hematopoietic cell kinases in immunohistochemical staining was assessed by H-score. Data are expressed as mean ± SEM. P < 0.05, P < 0.01, P < 0.001. P < 0.05 is considered statistically significant.

[0020] Figures 3A-3EKnockdown of hematopoietic cell kinase inhibited the proliferation of endometriosis stromal cells and promoted their apoptosis. Endometriosis stromal cells were transiently transfected with a non-targeting siRNA control (control group) and a siRNA targeting hematopoietic cell kinase (siHCK). Quantitative PCR and immunofluorescence staining (using an anti-hematopoietic cell kinase antibody) were used to detect the mRNA and protein expression of hematopoietic cell kinase in the transfected cells, respectively. Figure 3A , 3B The figure shows representative images of transfected cells stained with anti-hematopoietic cell kinase antibody. The images were obtained using the CCK8 assay. Figure 3C ) and Ki67 immunofluorescence staining ( Figure 3D The viability and proliferative capacity of transfected cells were assessed. TUNEL assays were also performed on the transfected cells. Figure 3E The apoptotic status of cells was detected. A bar chart shows the average percentage of apoptotic cells. Data are expressed as mean ± SEM. P < 0.05, P < 0.01. P < 0.05 is considered statistically significant.

[0021] Figure 4A-4L Knocking down hematopoietic cell kinases inhibited endometriosis in vivo. Figure 4A This example illustrates the timeline of establishing an in vivo endometriosis model and siRNA therapy to study the action of hematopoietic cell kinases. Figure 4B To assess the safety of siRNA treatment, mouse body weight was measured before treatment (T0) and after treatment (T7). This was achieved via subcutaneous... Figure 4C-4G ) and abdominal cavity ( Figure 4H-4L An endometriosis mouse model (n=5) was used to evaluate the effect of HCK knockdown on endometriosis. Figure 4C , 4H The volume and weight of the resected lesions were recorded and compared. The mRNA and protein expression of hematopoietic cell kinases in mouse endometriosis xenografts were analyzed by quantitative PCR and immunohistochemistry (IHC). Figure 4D-4E , Figure 4I-4J Representative images of paraffin-embedded mouse xenografts with hematopoietic kinase (HKK) stained with anti-hematopoietic cell kinase antibody are shown. The proliferation of xenografts treated with siRNA targeting HKKK was analyzed by Ki-67 immunohistochemical staining and TUNEL assay. Figure 4F , 4K ) and apoptosis ( Figure 4G (4L) state. Representative images for each experimental group are shown. Data are expressed as mean ± SEM. P < 0.05, P < 0.01. P < 0.05 is considered statistically significant.

[0022] Figure 5 The potential of hematopoietic kinases as immune targets in endometriosis. The figure illustrates the dysregulated pathways associated with hematopoietic kinases in endometriosis. These pathways were annotated using the Reactome database, and the degree of dysregulation was determined using the iPANDA algorithm. Green and red bars represent the number of comparisons of significant activation and inactivation of the corresponding pathways, respectively. Pathways marked in blue are related to immunity.

[0023] Figure 6 Abnormal expression of hematopoietic cell kinases is observed in the endometriosis proteomics dataset. Box plot of protein expression of hematopoietic cell kinases in the PDX006553 endometriosis proteomics dataset obtained from the ProteomeXchange Consortium. Invention Details

[0025] It should be understood that the present invention is not limited to the specific embodiments described herein. It should also be understood that the terminology used herein is for describing specific embodiments only and is not intended to be limiting, as the scope of the invention is defined only by the appended claims.

[0026] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. While any methods and materials similar to or equivalent to those described herein may be used to practice or test the invention, preferred methods and materials will be described below. All publications mentioned herein are for the purpose of disclosure and description of methods and / or materials in connection with reference to those publications.

[0027] If a given range of values ​​contains one or two boundaries, it should be understood that the smaller range between any intermediate value within the given range and either boundary of the range is also included in this invention. If a given range contains one or two boundaries, the range excluding one or both boundaries is also included in this invention.

[0028] the term

[0029] It must be noted that, unless the context clearly specifies otherwise, the singular forms “a,” “an,” and “the” used herein and in the appended claims include plural references. It should also be noted that claims may be drafted to exclude any optional elements. Therefore, this statement is intended to provide a basis for the use of exclusive terms such as “only,” “unique,” ​​or negative qualifiers in the description of claim elements.

[0030] Unless otherwise stated, the terms “comprising,” “including,” “containing,” and variations thereof do not imply the exclusion of other members, components, integers, or steps. These terms also include the meaning of “consisting of.” The term “consisting of” is a specific embodiment of the term “comprising,” in which any other unstated member, component, integer, or step is excluded.

[0031] The term "about" refers to a range equal to plus or minus 10 percent (+ / - 10%) of a specific value.

[0032] The term “and / or” refers to any one, several or all of the elements connected by the term.

[0033] The term “endometriosis” as used in this article refers to the abnormal presence of tissue containing typical endometrial granules and stromal components in the pelvic cavity or other parts of the body (most commonly the abdominal cavity).

[0034] As used herein, the term "treatment" refers to the relief, suppression, and / or reversal of disease progression (e.g., endometriosis). The term "treatment" includes any marker of successful treatment or improvement of the disease, including any objective or subjective parameters such as reduction; remission; symptom relief or increased tolerance of the injury, pathological condition, or symptom to an individual; delay or slowing of the rate of progression, etc. The determination of treatment or improvement may be based on the results of physical examinations, pathological tests, and / or diagnostic tests known in the art, for example. Treatment may also refer to a reduction in the incidence or flare rate of the disease, or its recurrence rate (e.g., prolonged remission time), compared to inaction. Clinically, such treatment may also be referred to as prevention.

[0035] As used in this article, the term "active agent" refers to a pharmaceutically active chemical substance that provides some pharmacological action and is used to treat or prevent diseases such as endometriosis.

[0036] The terms “inhibitor” and “antagonist” as used herein are used interchangeably and refer to any molecule that partially or completely blocks or inhibits the activity of a target (e.g., a protein used as a target in this invention). The terms “inhibitor” and “antagonist” as used herein may also refer to “activator”.

[0037] As used herein, the term "derivative" of a compound means any pharmaceutically acceptable molecule derived from (i.e., structurally related to) the compound and having similar or substantially the same activity as the compound, which, when administered to an individual, can (directly or indirectly) provide an active agent or its active metabolite. Examples of derivatives include, but are not limited to, pharmaceutically acceptable salts, hydrates, solvates, prodrugs, or metabolites.

[0038] As used herein, the term "pharmaceutically acceptable salt" refers to a relatively non-toxic inorganic or organic acid salt of the compounds of this invention. These salts can be prepared in situ during the final separation and purification of the compounds, or by reacting the purified compound in its free form alone with a suitable organic or inorganic acid and separating the resulting salt. Representative acid salts include, but are not limited to, acetates, adipates, aspartates, benzoates, benzenesulfonates, bicarbonates / carbonates, bisulfates / sulfates, borates, camsylates, citrates, cyclamates, ethanedisulfonates, ethanesulfonates, formates, fumarates, gluceptates, gluconates, glucuronates, hexafluorophosphates, and hydroxybenzoylbenzene salts. (benzate), hydrochloride / chloride, hydrobromide / bromide, hydroiodide / iodide, hydroxyethyl sulfonate, lactate, malate, maleate, malonate, methanesulfonate, methyl sulfate, naphthalate, 2-naphthalenesulfonate, nicotinate, nitrate, orotate, oxalate, palmitate, pamoate, phosphate / hydrogen phosphate / dihydrogen phosphate, pyroglutamate, glycosidate, stearate, succinate, tannic acid salt, tartrate, toluenesulfonate, trifluoroacetate, and xinafoate salt. In one embodiment, the pharmaceutically acceptable salt is a hydrochloride / chloride salt.

[0039] As used herein, the term "solvent" refers to a complex of variable stoichiometry formed by a solute (e.g., the active agent of the present invention) and a solvent. Such a solvent used for the purposes of the present invention may not interfere with the biological activity of the solute. Examples of suitable solvents include, but are not limited to, water, methanol, ethanol, and acetic acid.

[0040] As used herein, the term "prodrug" refers to a precursor that, when applied to a biological system, produces the compound as a product. For example, a prodrug may have the structure X-drug, where X is an inert carrier moiety and the drug is the active compound.

[0041] As used herein, the term "metabolite" refers to a molecule produced by modifying or processing a compound after administration to an individual. The term "metabolite" can also refer to a modified or processed drug that retains at least some of the activity of the parent compound.

[0042] As used herein, the term “pharmaceutically acceptable” means those compounds, materials, compositions, and / or dosage forms that, within reasonable medical judgment, are suitable for contact with individual tissues without excessive toxicity, irritation, allergic reactions, or other problematic complications commensurate with a reasonable benefit / risk ratio.

[0043] As used in this article, the term "pharmacologically acceptable carrier" refers to any carrier that, when administered to an individual, has substantially no long-term or permanent harmful effects, such as stabilizers, diluents, additives, adjuvants, excipients, etc. A "pharmacologically acceptable carrier" should be a pharmaceutically inert material that is substantially non-biologically active and constitutes an important part of the formulation.

[0044] As used herein, the term "individual" means any organism to which the active agent of the compositions of the present invention may be administered, for example for experimental, diagnostic, preventive, and / or therapeutic purposes. Typical individuals include animals (e.g., mammals such as mice, rats, rabbits, non-human primates such as chimpanzees and other ape and monkey species, and humans). Individuals can be mammals, particularly humans, including males or females, and include newborns, infants, juveniles, adolescents, adults, or the elderly, as well as different races and ethnicities.

[0045] As used herein, the terms "therapeutic effective dose" or "effective dose" are used interchangeably with "therapeutic effective amount" or "effective amount," referring to an amount effective in treating a disease (e.g., endometriosis) as recorded through clinical testing and evaluation, patient observations, etc. "Effective amount" may further specify the amount that results in a detectable change in biological or chemical activity. Those skilled in the art can detect and / or further quantify the detectable change in relation to the relevant mechanism or process. Furthermore, "effective amount" may specify the amount that maintains a desired physiological state, i.e., reduces or prevents a significant decline in the condition and / or promotes improvement in the condition.

[0046] As used herein, the term "unit dosage form" refers to a physically dispersed unit (e.g., capsule, tablet, or loaded syringe) suitable as an individual unit dose, each unit containing a predetermined amount of active agent calculated in conjunction with the desired pharmaceutical carrier to produce the desired therapeutic effect.

[0047] As used herein, the term "unit dose" refers to the dose of a substance (e.g., the active agent of the present invention) in a unit dosage form.

[0048] Active agent used to treat endometriosis

[0049] Through extensive research, the inventors have discovered a target for the treatment of endometriosis: hematopoietic cell kinase (HCK). Antagonists or agonists of this target can be used to treat endometriosis.

[0050] Hematopoietic kinases (SFKs) are members of the SRC family of cytoplasmic tyrosine kinases and are expressed in myeloid and B-lymphocyte lineage cells. Excessive SFK activation is associated with several types of leukemia and enhances cell proliferation and survival through physical binding to oncogenic fusion proteins and functional interactions with receptor tyrosine kinases. Elevated SFK activity has also been observed in many solid malignancies, including breast and colon cancer, and is associated with decreased patient survival. SFKs enhance the secretion of growth factors and pro-inflammatory cytokines by myeloid cells and promote alternating activation phenotypic polarization of macrophages towards wound healing and tumor promotion.

[0051] The active agent used to treat endometriosis can be any of the hematopoietic cell kinase inhibitors, including but not limited to posutinib, dasatinib, KIN-8194, RV568, rabastinibril and their derivatives.

[0052] The structure of an exemplary inhibitor of hematopoietic cell kinase is shown below.

[0053]

[0054] The amount of each active agent in a unit dosage form can be 1-1000 mg, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 50, 60, 70, 75, 80, 90, 100, 110, 120, 125, 130, 140, 150, 160, 170, 175, 180, 190, 200, 250, 300, 350, 400, 450, 500, 600, 700, 750, 800, 900, 1000 mg or any range between any two of the above specific values.

[0055] application

[0056] Each active agent of the present invention can be administered to an individual via oral, sublingual, rectal, vaginal, parenteral, intradermal, or nasal routes. Parenteral administration includes intravenous, intraperitoneal, intradermal, subcutaneous, intramuscular, intracranial, intrathecal, intratumoral, intratumoral, transdermal, transarticular, intramural, intrasynovial, intrathecal, intrahepatic, intralesional, or intracranial injection or infusion.

[0057] The active agents used herein can be formulated into pharmaceutical compositions for administration using known techniques. See, for example, Remington, The Science and Practice of Pharmacy (9th edition, 1995). In the preparation of pharmaceutical compositions according to the invention, the active agent is typically mixed, particularly with a pharmaceutically acceptable carrier. Of course, the carrier must be acceptable in the sense of compatibility with any other component in the formulation and must not be harmful to the patient. The carrier can be solid or liquid, or both, and is preferably formulated with the compound into a unit-dose formulation, such as a tablet, which may contain 0.01% or 0.5% to 95% or 99% of the active agent by weight. One or more active agents can be incorporated into the formulations of the invention, which can be prepared by any well-known pharmaceutical technique, including mixing components, optionally including one or more excipients and / or excipients. In some embodiments, any composition, carrier, excipient, excipient, and / or formulation of the invention comprises an ingredient from a natural or non-natural source. In other embodiments, any component of the composition, carrier, excipient, excipient, and / or formulation of the invention may be provided in a sterile form. Non-limiting examples of sterile carriers include endotoxin-free water or pyrogen-free water.

[0058] In some embodiments, the pharmaceutical compositions of the present invention are provided as part of a sterile composition / preparation, comprising the active agent of the present invention and a pharmaceutically acceptable carrier and / or excipient.

[0059] Suitable dosage forms for oral administration include tablets, capsules, powders, pills, granules, suspensions, solutions or solution preconcentrates, emulsions or emulsion preconcentrates. Pharmaceutically acceptable carriers that can be used in oral dosage forms include water, glycols, oils, alcohols, flavoring agents, preservatives, and coloring agents. Carriers such as starch, sugars, microcrystalline cellulose, diluents, fillers, flow aids, granulating agents, lubricants, binders, stabilizers, and disintegrants can be used to prepare oral solid dosage forms, such as powders, capsules, or tablets.

[0060] Diluents include, but are not limited to, microcrystalline cellulose, mannitol, powdered sugar, compressible sugar, dextran, dextrin, spinose, lactose, cellulose powder, sorbitol, sucrose, and talc, or combinations thereof. Based on the total weight of the oral composition, the diluent can be 5% to 90%, preferably 10% to 80%, 20% to 70%, 30% to 60%, or 40% to 50%.

[0061] Disintegrants include, but are not limited to, cellulose, alginate, gum, cross-linked polymers such as crospovidone or crospovidone, crospovidone sodium carboxymethyl cellulose, crospovidone calcium carboxymethyl cellulose, soybean polysaccharide, sodium starch glycolate, guar gum, or any combination thereof. The disintegrant may be present in an amount of about 1% to 15%, preferably 2% to 10%, based on the total weight of the oral composition.

[0062] The binder includes, but is not limited to, starch, cellulose or derivatives thereof, such as microcrystalline cellulose, hydroxypropyl cellulose, hydroxyethyl cellulose and hydroxypropyl methyl cellulose, sucrose, glucose, corn syrup, polysaccharides, gelatin or any combination thereof. Based on the total weight of the composition, the binder may be present in an amount of about 0.01 to 10%, preferably 1% to 10%.

[0063] Gliding agents include, but are not limited to, colloidal silica, magnesium trisilicate, cellulose powder, talc, or combinations thereof. Based on the total weight of the composition, the gliding agent may be present in an amount of 0.1% to 10%, preferably 0.1% to 0.5%.

[0064] Dosage forms may be, for example, tablets or capsules, and the effective dose may be provided in the form of one or more tablets, capsules, etc., and may be provided once daily or throughout the day at intervals of, for example, 4, 8, or 12 hours. For example, tablets or capsules may contain, for example, 10, 25, 50, 75, 100, 150, 200, 250, 300, 350, 400, 450, 500, 600, 700, 800, 900, 1,000, 1,100, or 1,250 mg of active agent. For example, administration of the active agent of the present invention to a human individual may include a daily dose in the range of 100-1,250, 150-1,000, 200-800, or 250-750 mg, which may be administered entirely once daily or in portions throughout the day. Liquid formulations may also be prepared so that any dose can be easily and conveniently dispensed.

[0065] Parenteral dosage forms are preferably sterile or capable of being sterilized before administration to an individual. Examples of parenteral dosage forms include, but are not limited to, injectable solutions, dried products that can be dissolved or suspended in pharmaceutically acceptable injectable carriers, injectable suspensions, and emulsions.

[0066] Suitable carriers that can be used to provide the parenteral dosage forms provided herein include, but are not limited to: water for injection; aqueous media, such as, but not limited to, sodium chloride injection, Ringer's solution, and glucose injection; water-miscible carriers, such as, but not limited to, ethanol, polyethylene glycol, and polypropylene glycol; and non-aqueous carriers, such as, but not limited to, corn oil, cottonseed oil, peanut oil, sesame oil, ethyl oleate, isopropyl myristate, and benzyl benzoate.

[0067] Compounds that increase the solubility of one or more of the active agents disclosed herein may also be incorporated into the parenteral dosage forms provided herein. For example, cyclodextrins and their derivatives can be used to improve the solubility of the active agents of the present invention.

[0068] It should be understood that the effective therapeutic dose can be determined by the physician based on factors such as the type, stage and / or severity of the disease, the individual's condition, age, weight, sex and response, and the route of administration.

[0069] The therapeutically effective dose is such that, when administered to an individual, it is sufficient to achieve plasma concentrations of about 0.01 μg / ml to about 100 μg / ml, about 0.1 μg / ml to about 10 μg / ml, and about 1 μg / ml to 5 μg / ml.

[0070] When the active agents of the present invention are administered to an individual, the therapeutically effective amount of each active agent of the present invention can generally be in the range of about 0.5 to about 250 mg / kg, about 1 to about 250 mg / kg, about 2 to about 200 mg / kg, about 3 to about 120 mg / kg, about 5 to about 250 mg / kg, about 10 to about 200 mg / kg, or about 20 to about 120 mg / kg. In some implementations, the effective therapeutic dose may be 0.5 mg / kg, 1 mg / kg, 2 mg / kg, 3 mg / kg, 4 mg / kg, 5 mg / kg, 6 mg / kg, 8 mg / kg, 10 mg / kg, 20 mg / kg, 25 mg / kg, 40 mg / kg, 50 mg / kg, 60 mg / kg, 75 mg / kg, 100 mg / kg, 120 mg / kg, 150 mg / kg, 175 mg / kg, 200 mg / kg, 225 mg / kg, 250 mg / kg, or 300 mg / kg.

[0071] Each active agent of the present invention may be applied once or twice daily; or once every 2, 3, 4, 5, 6, 7, 8, 9, or 10 days; or once every 1, 2, or 3 weeks. In some embodiments, each active agent of the present invention may be applied in a five-times-a-week regimen. In a five-times-a-week regimen, it may be applied for five consecutive days (once a day), followed by two consecutive days of rest.

[0072] As used herein, the term "kit" refers to packaging and, generally, instructions for use. The active agent or pharmaceutical composition in a kit may be in any of a variety of forms suitable for distribution in the kit. Such forms may include liquids, powders, tablets, suspensions, etc. Two or more active agents may be provided in separate containers suitable for individual administration, or in a composition in a single container within the packaging. Depending on the treatment method, the kit may contain an amount sufficient for a dose of one or more active agents. Instructions for use typically contain written instructions on how to treat a disease (e.g., ALS) with the active agent in the kit.

[0073] It should be understood that, in addition to the active agent of the present invention, the combination or pharmaceutical composition of the present invention may include other therapeutic agents or therapies, such as biological therapeutic agents and / or chemotherapeutic agents. In addition to applying the active agent of the present invention, the method may include applying other therapeutic agents or therapies, such as biological therapeutic agents and / or chemotherapeutic agents. Other therapeutic agents or therapies may be applied simultaneously, alone, or sequentially with the therapeutic agents of the present invention.

[0074] Example

[0075] Example 1. Using PandaOmics TM -AI-enabled biological target discovery platform for identifying intrauterine targets Therapeutic targets and drug reuse candidates for membranes

[0076] 1. Method

[0077] Data sources and availability

[0078] The current analysis utilized batch and single-cell transcriptomics and proteomics data. A total of 36 endometriosis-related batch transcriptomics datasets (including microarrays and RNA sequencing series) from various tissue sources retrieved from GeneExpression Omnibus (GEO) and ArrayExpress were available for direct downstream analysis and target identification using PandaOmics.

[0079] Datasets and Comparison Selection

[0080] Since the pathogenesis of the disease begins with the situ endometrium, endometrial tissue datasets were selected for downstream analysis and target identification. Eleven case-control comparisons were generated from the eleven batch transcriptomics datasets listed in Table 1. Normal endometrial tissue from healthy individuals and situ endometrial tissue from patients with endometriosis were selected as controls, labeled "healthy" and "situ endometrium" respectively in the experimental design column of Table 1.

[0081] Table 1. Meta-analysis comparison of endometriosis for target selection

[0082]

[0083] PandaOmics target identification

[0084] PandaOmics is a cloud-based target discovery platform that combines multiple deep learning models and AI algorithms in the target prioritization process. Twenty-three target prioritization models have been developed, covering omics (Omics), text-based, financial, and key opinion leader (KOL) data, to predict the association between target genes and specific indications. Each model is scored on a normalized scale from zero to one, with higher scores corresponding to better predicted target-disease associations. These models are validated using time-machine methods to demonstrate their capabilities in target identification. The target list can also be refined using adjustable filters based on drugability, tissue specificity, target family, and development status.

[0085] To identify potential targets for endometriosis, a case-control batch transcriptomics comparison was assigned to a meta-analysis. To identify actionable targets with varying levels of novelty, filter and scoring settings were customized to re-prioritize targets. Only targets belonging to druggable protein classes were retained in the analysis, excluding those considered essential genes as defined by the Therapeutic Target Database (TTD). Therefore, three lists of druggable targets were generated: high confidence, moderate novelty, and high novelty, with the top 50 targets undergoing further analysis.

[0086] Experimental validation of potential targets

[0087] After identifying potential therapeutic targets based on PandaOmics and expression analysis, immunohistochemical (IHC) analysis was performed on targets in paraffin-embedded ovarian endometrial stroma (EMS), peritoneal EMS, and normal EMS tissues to confirm their protein expression using anti-hematopoietic cell kinase antibodies.

[0088] First, functional analysis of the confirmed target was performed in EMS cells using siRNA knockdown. EMS cells were transfected with 10 nM of non-targeting control siRNA or siRNA targeting hematopoietic kinase, and hematopoietic kinase expression was confirmed by quantitative PCR (qPCR) and immunofluorescence (IF) staining with anti-hematopoietic kinase antibody. Three functional assays were then performed, including CCK8 cell viability assay, IF staining for cell proliferation with anti-Ki-67 antibody, and TUNEL assay for cell apoptosis. CCK8 and TUNEL assays were performed according to the manufacturer's instructions.

[0089] An endometriosis-mimicking mouse model was established by subcutaneously and intraperitoneally transplanting endometrial tissue excised from the uterine horn of a healthy female mouse into another healthy female mouse. Two weeks after transplantation, the mice were divided into two groups and received either treatment: intraperitoneal administration of hematopoietic cell kinase siRNA or a non-targeted siRNA control, for one week. Mouse weight was monitored daily. On day 7, the mice were sacrificed and the peritoneal lesions were excised. Lesion volume and weight were recorded. Hematopoietic cell kinase expression in the xenograft was validated by qPCR and IHC, followed by Ki-67 IHC staining and TUNEL assay.

[0090] 2. Results

[0091] Identifying hematopoietic cell kinases as potential therapeutic targets using PandaOmics

[0092] Based on expression correlation results, target identification was performed using a meta-analysis of endometriosis with 11 subtypes and cycle nonspecific comparisons (Table 1). To identify potential therapeutic targets for endometriosis, 141 unique genes with three novelty levels were screened based on rankings calculated by PandaOmics, consistency of dysregulated expression in comparisons included in the meta-analysis, statistical significance of dysregulation, and literature support indicating potential roles in driving or promoting endometriosis. Hematopoietic cell kinase was identified as a potential therapeutic candidate for endometriosis. In the meta-analysis, it was ranked among the top 50 targets under the novelty setting and had 6 omics scoring models, with 6 models scoring 0.8 or higher. Figure 1B ).

[0093] Sustained upregulation of hematopoietic cell kinases in endometriosis samples

[0094] To investigate the functional correlation between hematopoietic cell kinases and endometriosis, the expression profiles of endometrial samples collected from endometriosis lesions and control endometrium were analyzed. Regarding batch transcriptomic characteristics, in 11 comparisons, 8 hematopoietic cell kinases were upregulated, with 3 of these upregulations reaching statistical significance (FDR < 0.05). Figure 2A ).

[0095] Immunohistochemical (IHC) staining was performed to assess the dysregulation of target protein expression in paraffin-embedded ovarian and peritoneal EMS tissue samples, revealing that hematopoietic cell kinases were significantly overexpressed in ovarian EMS tissue compared to normal endometrial stroma. Figure 2B and 2CWe also observed dysregulation of the target expression in three endometriosis proteomics datasets. Hematopoietic cell kinase was observed in one of the comparisons. Although not statistically significant, hematopoietic cell kinase was upregulated in endometriosis samples compared to healthy controls. Figure 6 Overall, consistent upregulation of hematopoietic cell kinases was observed at both the mRNA and protein levels in endometriosis samples, supporting their potential to drive endometriosis.

[0096] Knocking down hematopoietic cell kinases reduces endometriosis both in vivo and in vitro.

[0097] To characterize the functional role of hematopoietic cell kinase (siHCK) in endometriosis, siHCK was knocked down in endometriotic stromal cells via transient transfection with siRNA. Quantitative PCR (qPCR) analysis and immunofluorescence (IF) staining both confirmed the silencing of siHCK in the cells. Figure 3A and 3B Functionally, knockdown of hematopoietic cell kinase significantly reduces the viability of transfected stromal cells. Figure 3C ) and the immunofluorescence intensity of Ki-67 ( Figure 3D Furthermore, knocking down hematopoietic cell kinases increased the percentage of apoptotic cells by at least 3-fold. Figure 3E ).

[0098] Functional characterization of hematopoietic cell kinases in a mouse model of endometriosis was performed in a similar manner. After successful subcutaneous and intraperitoneal implantation of endometrial tissue, mice were treated with siRNA for one week. Figure 4A As reflected in the steady increase in body weight, treatment with siHCK has been shown to have no harmful effects in mice. Figure 4B In contrast to those treated with control siRNA, the volume and weight of subcutaneous lesions treated with siHCK were significantly reduced. Figure 4C Subcutaneous lesions treated with siHCK showed that hematopoietic cell kinases were present in mRNA ( Figure 4D ) and protein levels ( Figure 4E The expression of Ki-67 IHC on the surface was reduced. Consistent with in vitro results, siHCK treatment reduced the staining intensity of Ki-67 IHC in endometriotic cells. Figure 4F Inhibition of hematopoietic cell kinases also significantly promoted apoptosis. Figure 4G Similar to subcutaneous treatment, intraperitoneal treatment with HCK significantly reduced the growth of ectopic lesions. Figure 4H The target knockdown of siHCK has been confirmed. Figure 4I siHCK tends to reduce the proliferative capacity of endometriotic cells. Figure 4K), and significantly induced apoptosis in ectopic lesions ( Figure 4L Considering both in vitro and in vivo results, inhibition of hematopoietic cell kinases demonstrated a preventive effect against endometriosis by inhibiting cell proliferation and activating apoptosis, suggesting its potential as a therapeutic agent for endometriosis.

[0099] Hematopoietic cell kinases as immune targets of endometriosis

[0100] Following functional characterization, dysregulation pathway analysis was performed to address the mechanistic explanation of the effects of hematopoietic cell kinases on endometriosis. Figure 5 As shown, the number of comparisons for significant dysregulation in each pathway (P<0.05) is represented by the length of the bars, where green and red represent activation and inhibition, respectively. Most of the altered pathways associated with hematopoietic kinases are related to immunity and infection (see figure below). Four of the eight hematopoietic kinase-related immune pathways (marked in blue) were activated. These findings highlight the potential for developing hematopoietic kinases as immune targets.

Claims

1. A method for treating endometriosis in individuals in need, comprising administering an effective amount of a hematopoietic cell kinase inhibitor to said individual.

2. The method according to claim 1, wherein the hematopoietic cell kinase inhibitor is one or more selected from bosutinib, dasatinib, KIN-8194, RV568, rabastinibril and their derivatives.

3. Use of hematopoietic cell kinase inhibitors in the preparation of drugs for the treatment of endometriosis.

4. The use according to claim 3, wherein the hematopoietic cell kinase inhibitor is one or more selected from bosutinib, dasatinib, KIN-8194, RV568, rabastinibril and their derivatives.

5. Medications used to treat endometriosis, wherein the medication contains a hematopoietic cell kinase inhibitor.

6. The medicament for the stated use according to claim 5, wherein the hematopoietic cell kinase inhibitor is one or more selected from bosutinib, dasatinib, KIN-8194, RV568, rabastinibril and their derivatives.

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