Pharmaceutical composition comprising prucalopride for preventing or treating intestinal damage caused by radiation treatment

Abstract

The present invention relates to a pharmaceutical composition for preventing or treating intestinal damage caused by radiation treatment, comprising prucalopride or a pharmaceutically acceptable salt, hydrate, or co-crystal thereof. The composition according to the present invention repairs damaged cell barrier function caused by radiation treatment and exhibits a protective effect against the loss of intercellular junction protein expression, thereby being able to be usefully employed as an adjuvant therapy in radiation therapy for the lower abdomen including the abdomen, pelvis, and rectum.

Description

Pharmaceutical composition for the prevention or treatment of intestinal damage caused by radiation treatment containing prucalopride

[0001] The present invention relates to a pharmaceutical composition for the prevention or treatment of intestinal damage caused by radiation treatment, comprising prucalopride or a pharmaceutically acceptable salt, hydrate, or co-crystal thereof, and a food composition for the prevention or improvement of intestinal damage caused by radiation treatment.

[0002] Radiation therapy is one of the three major cancer treatments, along with surgery and chemotherapy. With the development of specialized radiation therapies, it is increasingly used in conjunction with other conventional cancer treatments. However, as radiation therapy becomes more widespread, the number of patients experiencing side effects is also rising. In particular, for cancers located in the lower abdomen—such as uterine, ovarian, bladder, and rectal cancers—intestinal inflammatory diseases can occur due to radiation exposure during the treatment process. Consequently, patients may experience symptoms such as nausea, vomiting, cramps, abdominal pain, diarrhea, bloody stools, and weight loss. While most symptoms generally disappear naturally after radiation therapy is completed, there is also a possibility that they may persist for months or years due to scarring or strictures within the intestines.

[0003] Unlike general inflammatory bowel disease, radiation-induced enteropathy involves oxidative stress caused by radiation inducing apoptosis in intestinal epithelial cells and damaging intercellular junctions, leading to intestinal tissue damage and impaired barrier function. Consequently, external bacteria and other pathogens enter the body, causing severe inflammation throughout the system. Currently, there are no FDA-approved medications for radiation-induced enteropathy, and only symptomatic treatment is provided. As radiation therapy and its associated side effects become more widespread, there is an urgent need to develop therapeutic substances capable of not only suppressing inflammation but also promoting regeneration or alleviating barrier damage.

[0004] Meanwhile, other drugs known as 5-HT4 receptor agonists, such as mosapride, have some effect in improving gastrointestinal motility, but have not shown significant therapeutic effects in regenerating intestinal tissue destroyed by radiation or restoring intestinal barrier function. Therefore, there is an urgent need to select specific drugs capable of controlling the fundamental pathological mechanisms of radiation-induced intestinal damage (inflammation, stem cell death, etc.) beyond simple 5-HT4 receptor stimulation, and to elucidate their mechanisms of action.

[0005] The object of the present invention is to provide a pharmaceutical composition and a food composition for preventing or treating intestinal damage caused by radiation treatment, a method for treating intestinal damage caused by radiation treatment using the same, and uses thereof.

[0006] Accordingly, the present invention provides a pharmaceutical composition for the prevention or treatment of intestinal damage caused by radiation treatment, comprising prucalopride or a pharmaceutically acceptable salt thereof.

[0007] In addition, the present invention provides a food composition for preventing or improving intestinal damage caused by radiation treatment, comprising prucalopride or a food-grade acceptable salt, hydrate, or co-crystal thereof.

[0008] In addition, the present invention provides a method for treating intestinal damage caused by radiation treatment, comprising the step of administering prucalopride or a pharmaceutically acceptable salt, hydrate, or co-crystal thereof to a subject requiring it.

[0009] In addition, the present invention provides the use of prucalopride of the present invention or its pharmaceutically acceptable salts, hydrates, and co-crystals for manufacturing medicines for the prevention or treatment of intestinal damage caused by radiation treatment.

[0010] In addition, the present invention provides a use of a composition comprising prucalopride or a pharmaceutically acceptable salt, hydrate, or co-crystal thereof for use in the prevention or treatment of intestinal damage caused by radiation treatment.

[0011] Hereinafter, the present invention will be described in detail with reference to the attached drawings and embodiments thereof. However, the following embodiments are presented as examples of the present invention, and if it is determined that a detailed description of a technology or configuration well known to a person skilled in the art may unnecessarily obscure the essence of the present invention, such detailed description may be omitted, and the present invention is not limited thereby. The present invention is capable of various modifications and applications within the scope of the claims set forth below and the equivalent scope interpreted therefrom.

[0012] Furthermore, the terminology used in this specification is used to appropriately describe preferred embodiments of the present invention, and may vary depending on the intent of the user or operator, or the conventions of the field to which the present invention belongs. Accordingly, the definitions of these terms should be based on the content throughout this specification. Throughout the specification, when a part is described as "comprising" a certain component, this means that, unless specifically stated otherwise, it does not exclude other components but may include additional components.

[0013] All technical terms used in this invention, unless otherwise defined, are used in the sense generally understood by a person skilled in the art related to this invention. Additionally, while preferred methods or samples are described herein, similar or equivalents are also included within the scope of this invention. The contents of all publications cited as references in this specification are incorporated into this invention.

[0014] In one embodiment for achieving the above objective, the present invention provides a pharmaceutical composition for the prevention or treatment of intestinal damage caused by radiation treatment, comprising prucalopride or a pharmaceutically acceptable salt thereof.

[0015] In addition, the present invention provides a veterinary composition for the prevention or treatment of intestinal damage caused by radiation treatment, comprising prucalopride or a pharmaceutically acceptable salt thereof.

[0016] The term "Prucalopride" used in the present invention may be a compound represented by the following chemical formula 1.

[0017] [Chemical Formula 1]

[0018]

[0019] The above chemical formula 1 is named 4-amino-5-chloro-2,3-dihydro-N-[1-(3-methoxypropyl)-4-piperidinyl]-7-benzofurancarboxamide.

[0020] More preferably, a commercially available compound may be a (1:1) succinic acid addition salt of 4-amino-5-chloro-2,3-dihydro-N-[1-(3-methoxypropyl)-4-piperidinyl]-7-benzofurancarboxamide, which has prokinetic activity, that is, strong gastrointestinal motility-promoting activity.

[0021] Specifically, prucalopride facilitates both cholinergic and non-cholinergic non-adrenergic (NANC) excitatory neurotransmission and promotes colonic motility and defecation in animals. Prucalopride has no affinity for 5-HT2A and 5-HT3 receptors but is a potent and selective agonist of 5-HT4 receptors. Prucalopride induces massive contractions transmitted as peristaltic waves along the length of the colon, thereby exhibiting a significant motility-enhancing effect on the large intestine.

[0022] In the present invention, prucalopride may exist in the form of a pharmaceutically acceptable salt. In the present invention, "pharmaceutically acceptable salt" refers to a salt commonly used in the pharmaceutical industry, and includes, for example, inorganic ionic salts prepared with calcium, potassium, sodium, magnesium, etc.; inorganic salts prepared with hydrochloric acid, nitric acid, phosphoric acid, bromic acid, iodic acid, perchloric acid, sulfuric acid, etc.; organic salts prepared with acetic acid, trifluoroacetic acid, citric acid, maleic acid, succinic acid, oxalic acid, benzoic acid, tartaric acid, fumaric acid, manderic acid, propionic acid, lactic acid, glycolic acid, gluconic acid, galacturonic acid, glutamic acid, glutaric acid, glucuronic acid, aspartic acid, ascorbic acid, carboxylic acid, vanillic acid, and hydroiodic acid, etc. Sulfonates prepared from methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, and naphthalenesulfonic acid; amino acid salts prepared from glycine, arginine, and lysine; and amine salts prepared from trimethylamine, triethylamine, ammonia, pyridine, and picoline, etc., but the types of salts referred to in the present invention are not limited by these listed salts.

[0023] Preferably, it may be a free base, fumarate, succinate, hydrochloride, citrate, or oxalate, and more preferably, a free base, succinate, or citrate.

[0024] In addition, the prucalopride according to the present invention comprises not only pharmaceutically acceptable salts but also all possible solvates, hydrates, and co-crystals that can be prepared therefrom.

[0025] In the present invention, a solvate refers to a complex or crystalline form in which prucalopride or a salt thereof is bonded to solvent molecules by non-covalent intermolecular forces. When the solvent is water, it is specifically referred to as a hydrate. The solvate or hydrate includes both forms containing stoichiometric and non-stoichiometric amounts of solvent or water.

[0026] In the present invention, a co-crystal refers to a crystalline material formed through non-covalent bonding, such as hydrogen bonding, pi-stacking, and van der Waals forces, in which prucalopride (API) and other pharmaceutically acceptable molecules (co-formers) exist in defined stoichiometric ratios within a crystal lattice. The said co-crystal is included without limitation as long as it is in a form capable of improving the solubility, stability, and bioavailability of prucalopride.

[0027] In the present invention, the "radiation treatment" may be by radiation therapy.

[0028] The aforementioned "radiation therapy" generally refers to the use of ionizing radiation to kill cancer cells as part of anticancer therapy. Ionizing radiation is generated using X-rays, gamma rays, or charged particles (e.g., protons or electrons). Radiation therapy may be delivered by a device located outside the patient's body (external-external radiation therapy), by a source located inside the patient's body (internal radiation therapy or brachytherapy), or through systemic radioisotopes delivered intravenously or orally (systemic radioisotope therapy). Radiation therapy may be planned and administered in conjunction with imaging-based technologies, such as computed tomography (CT) and magnetic resonance imaging (MRI), to accurately determine the dose and location of the radiation to be administered. In various embodiments, the radiation therapy is selected from the group consisting of total all-body radiation therapy, conventional external radiation therapy, stereotactic radiosurgery, stereotactic body radiation therapy, 3-D conformal radiation therapy, intensity-modulated radiation therapy, image-guided radiation therapy, tomotherapy, brachytherapy, and total all-body radiation therapy. Depending on intent, in some embodiments, the radiation therapy is curative, adjuvinating, or palliative. In certain embodiments, the term "radiation therapy" refers to fractionated radiation therapy.

[0029] In the present invention, the "intestinal damage" may be one or more selected from the group consisting of inflammation, edema, tumor, ulcer, scar, and stenosis, and preferably may be intestinal damage caused by radiation treatment or radiation irradiation.

[0030] In the case of radiation-induced intestinal damage, it may occur when the intestines are exposed to radiation during radiation therapy, particularly for cancers located in the lower abdomen such as uterine cancer, ovarian cancer, bladder cancer, and rectal cancer. Unlike general inflammatory bowel disease, radiation-induced intestinal damage has a clear cause of onset, namely radiation. Oxidative stress caused by radiation damages the intercellular junctions of intestinal epithelial cells, leading to damage to intestinal tissue and dysfunction of the intestinal barrier. Consequently, external bacteria may enter the body, potentially causing severe inflammation throughout the body. According to one embodiment, radiation-induced intestinal damage may be radiation-induced enteropathy or radiation-induced enteritis.

[0031] In addition, if intestinal damage is induced by radiation treatment, changes in the expression of genes related to the length of intestinal villi or intestinal epithelial permeability, such as Villin, Cldn3 (claudin-3) and Zo-1, genes related to intestinal stem cells such as Olfm4 (olfactomedin-4) and Lgr5, and genes related to inflammation such as IL-1β, TNF-α, IL-6, and Mcp-1 may occur in the intestinal tissue.

[0032] According to one embodiment of the present invention, the composition according to the present invention has the effect of promoting intestinal motility in a mouse model in which intestinal damage caused by radiation treatment is induced, the effect of regenerating damaged intestinal tissue, and the effect of promoting the recovery rate of the length of intestinal villi.

[0033] In addition, according to one embodiment of the present invention, the composition according to the present invention has the effect of alleviating intestinal inflammation in a mouse model of intestinal damage induced by radiation treatment, and has the effect of suppressing changes in the expression of one or more genes selected from the group consisting of IL-1β, TNF-α, IL-6, and Mcp-1, which are genes related to the degree of inflammation.

[0034] In addition, according to one embodiment of the present invention, the composition according to the present invention has the effect of restoring damaged intestinal epithelial barrier function and protecting intestinal stem cells in a mouse model in which intestinal damage is induced by radiation treatment, and has the effect of promoting the expression of one or more genes selected from the group consisting of Villin, Cldn3 (claudin-3), and Zo-1, which are genes related to intercellular junction molecules and pericellular permeability of intestinal epithelial cells, and Olfm4 (olfactomedin-4) and Lgr5, which are genes related to intestinal stem cells.

[0035] That is, the pharmaceutical composition according to the present invention has the effect of promoting the expression of one or more genes selected from the group consisting of Villin, Olfm4 (olfactomedin-4), Lgr5, Cldn3 (claudin-3), and Zo-1.

[0036] In addition, the pharmaceutical composition according to the present invention has the effect of inhibiting the expression of one or more genes selected from the group consisting of IL-1β, TNF-α, IL-6, and Mcp-1.

[0037] In addition, the pharmaceutical composition according to the present invention has the effect of promoting the expression of one or more genes selected from the group consisting of Villin, Olfm4 (olfactomedin-4), Lgr5, Cldn3 (claudin-3), and Zo-1, and inhibiting changes in the expression of one or more genes selected from the group consisting of IL-1β, TNF-α, IL-6, and Mcp-1.

[0038] The term "prevention" as used in the present invention refers to any act of suppressing intestinal damage caused by radiation treatment through the administration of the composition. In the present invention, "treatment" refers to any act of improving or beneficially altering the symptoms of the said disease through the administration of the composition.

[0039] The pharmaceutical composition of the present invention may include a pharmaceutically acceptable carrier and may be formulated according to conventional methods into oral formulations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, and aerosols, as well as topical preparations, suppositories, and sterile injectable solutions.

[0040] The above pharmaceutically acceptable carriers include, but are not limited to, those commonly used in the art, such as lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil.

[0041] In addition, the pharmaceutical composition of the present invention may include, but is not limited to, fillers, extenders, binders, wetting agents, disintegrants, diluents or excipients such as surfactants, and other pharmaceutically acceptable additives.

[0042] When the pharmaceutical composition of the present invention is formulated into an oral solid dosage form, it includes tablets, pills, powders, granules, capsules, etc., and such solid dosage forms may include at least one excipient, such as starch, calcium carbonate, sucrose or lactose, gelatin, etc., and may include, but are not limited to, lubricants such as magnesium stearate, talc, etc.

[0043] When the pharmaceutical composition of the present invention is formulated into an oral liquid form, it includes suspensions, liquid formulations, emulsions, syrups, etc., and includes diluents such as water and liquid paraffin, humectants, sweeteners, flavorings, preservatives, etc., but is not limited thereto.

[0044] When the pharmaceutical composition of the present invention is formulated for parenteral use, it comprises a sterile aqueous solution, a non-aqueous solvent, a suspension agent, an emulsion, a lyophilized preparation, and a suppository. The non-aqueous solvent and suspension agent may include, but are not limited to, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. The base of the suppository may include, but is not limited to, Witepsol, Macrogol, Tween 61, cocoa gluten, laurin gluten, glycerogelatin, etc.

[0045] In another aspect of the present invention, the present invention provides a method for treating intestinal damage caused by radiation treatment, comprising the step of administering prucalopride or a pharmaceutically acceptable salt, hydrate, or co-crystal thereof to a subject in need thereof.

[0046] The pharmaceutical composition of the present invention may be administered to an adult (based on a body weight of about 60 kg) at a dose of about 0.1 mg to 100 mg per day, preferably 0.5 mg to 10 mg, based on prucalopride, which is the active ingredient. The above dose is set by converting the results of mouse experiments (5 mg / kg) into a human equivalent dose (HED) and considering the safety margin, but it may be appropriately increased or decreased under a doctor's prescription depending on the patient's condition and the severity of the disease.

[0047] The preferred dosage of the composition of the present invention varies depending on the patient's condition and weight, the severity of the disease, the form of the drug, the route of administration, and the duration, but can be appropriately selected by those skilled in the art. The said dosage does not limit the scope of the present invention in any way.

[0048] The pharmaceutical and / or veterinary compositions of the present invention may be administered to mammals, such as rats, mice, livestock, and humans, via various routes. In particular, they may be administered to humans, as well as to companion animals or laboratory animals undergoing radiation therapy, via various routes. All modes of administration are expected, for example, orally, rectally or intravenously, intramuscularly, subcutaneously, intradurally, or intracerebroventricularly, and the preferred mode of administration is easy and inexpensive oral administration.

[0049] Another aspect of the present invention provides the use of prucalopride of the present invention or a pharmaceutically acceptable salt thereof for manufacturing a medicine for the prevention or treatment of intestinal damage caused by radiation treatment.

[0050] Another aspect of the present invention provides a composition comprising prucalopride or a pharmaceutically acceptable salt thereof for use in the prevention or treatment of intestinal damage caused by radiation treatment.

[0051] Another aspect of the present invention provides the use of a composition comprising prucalopride or a pharmaceutically acceptable salt thereof for use in the prevention or treatment of intestinal damage caused by radiation treatment.

[0052] In one embodiment for achieving the purpose of the present invention, the present invention provides a food composition for preventing or improving intestinal damage caused by radiation treatment, comprising prucalopride or a pharmaceutically acceptable salt thereof.

[0053] In the present invention, the "radiation treatment" may be by radiation therapy.

[0054] In the present invention, the "intestinal damage" may be one or more selected from the group consisting of inflammation, edema, tumor, ulcer, scar, and stenosis, and preferably may be intestinal damage caused by radiation treatment or radiation irradiation.

[0055] In the case of radiation-induced intestinal damage, it may occur when the intestines are exposed to radiation during radiation therapy, particularly for cancers located in the lower abdomen such as uterine cancer, ovarian cancer, bladder cancer, and rectal cancer. Unlike general inflammatory bowel disease, radiation-induced intestinal damage has a clear cause of onset, namely radiation. Oxidative stress caused by radiation damages the intercellular junctions of intestinal epithelial cells, leading to damage to intestinal tissue and dysfunction of the intestinal barrier. Consequently, external bacteria may enter the body, potentially causing severe inflammation throughout the body. According to one embodiment, radiation-induced intestinal damage may be radiation-induced enteropathy or radiation-induced enteritis.

[0056] In the present invention, the food composition has the effect of promoting the expression of one or more genes selected from the group consisting of Villin, Olfm4 (olfactomedin-4), Lgr5, Cldn3 (claudin-3), and Zo-1.

[0057] In the present invention, the food composition has the effect of inhibiting the expression of one or more genes selected from the group consisting of IL-1β, TNF-α, IL-6, and Mcp-1.

[0058] In the present invention, the food composition has the effect of promoting the expression of one or more genes selected from the group consisting of Villin, Olfm4 (olfactomedin-4), Lgr5, Cldn3 (claudin-3), and Zo-1, and inhibiting the expression of one or more genes selected from the group consisting of IL-1β, TNF-α, IL-6, and Mcp-1.

[0059] The term "improvement" as used in the present invention refers to any action that at least reduces parameters related to the condition being treated, such as the degree of symptoms.

[0060] In the present invention, when the composition is used as a food composition, the composition may be added as is or used together with other food or food ingredients, and may be used appropriately according to conventional methods.

[0061] The above composition may include food-grade acceptable food additives in addition to the active ingredient, and the amount of the active ingredient can be appropriately determined according to the purpose of use (prevention, health, or therapeutic treatment).

[0062] The term "food auxiliary additive" as used in the present invention refers to a component that can be added to food as an auxiliary component, and can be appropriately selected and used by a person skilled in the art as it is added to manufacture health functional foods of each formulation. Examples of food auxiliary additives include various nutritional supplements, vitamins, minerals (electrolytes), flavoring agents such as synthetic flavoring agents and natural flavoring agents, coloring agents and fillers, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc., but the types of food auxiliary additives of the present invention are not limited by the above examples.

[0063] The food composition of the present invention may include a health functional food. The term "health functional food" as used in the present invention refers to a food manufactured and processed in the form of tablets, capsules, powders, granules, liquids, pills, etc., using raw materials or ingredients having functional properties useful to the human body. Here, "functionality" means obtaining effects useful for health purposes, such as regulating nutrients or physiological actions regarding the structure and function of the human body. The health functional food of the present invention may be manufactured by methods commonly used in the ordinary technical field, and may be manufactured by adding raw materials and ingredients commonly added in the ordinary technical field during such manufacturing. Furthermore, the formulation of the health functional food may also be manufactured without restriction as long as it is a formulation recognized as a health functional food.

[0064] The food composition of the present invention can be manufactured in various forms of formulations. Unlike general pharmaceuticals, it uses plants as raw materials and has the advantage of not causing side effects that may occur with long-term use. Additionally, due to its excellent portability, the health functional food of the present invention can be consumed as an adjuvant to enhance the effects of treatments for autoimmune diseases.

[0065] Food compositions to which prucalopride can be added include, for example, various foods, beverages, chewing gum, tea, vitamin complexes, health functional foods, etc.

[0066] The health functional beverage composition of the present invention contains the above-mentioned prucalopride as an essential ingredient in the indicated proportions, and there are no special restrictions on other ingredients; it may contain additional ingredients such as various flavorings or natural carbohydrates, as in conventional beverages.

[0067] Examples of the above natural carbohydrates include monosaccharides, e.g., glucose, fructose, etc.; disaccharides, e.g., maltose, sucrose, etc.; and polysaccharides, such as conventional sugars like dextrin, cyclodextrin, etc.; and sugar alcohols such as xylitol, sorbitol, erythritol, etc. As flavoring agents other than those mentioned above, natural flavoring agents such as thaumatin, stevia extract, e.g., rebaudioside A, glycyrrhizin, etc.; and synthetic flavoring agents, e.g., saccharin, aspartame, etc. may be advantageously used.

[0068] The present invention will be explained in more detail below through examples. The following examples are merely for the purpose of explaining the present invention more specifically, and it will be obvious to those skilled in the art that the scope of the present invention is not limited to these examples.

[0069] The composition of the present invention can protect against intestinal barrier damage caused by radiation treatment and inhibit the infiltration of intestinal inflammatory cells. Furthermore, it repairs damage to intestinal stem cells and intestinal epithelial cells caused by radiation treatment, promotes intestinal motility, and exhibits a protective effect against the loss of expression of intercellular junction proteins, thereby demonstrating a preventive or therapeutic effect against intestinal damage caused by radiation treatment.

[0070] In particular, the present invention confirmed that the therapeutic effects of prucalopride (intestinal motility, tissue regeneration, anti-inflammation, and restoration of barrier function) are offset when GR-113808, a 5-HT4 receptor selective antagonist, is co-administered; thereby pharmacologically clarifying that these therapeutic effects are attributed to the activation of 5-HT4 receptor-mediated signaling pathways by prucalopride. This demonstrates that the composition of the present invention fundamentally treats radiation-induced intestinal damage through a clear mechanism of action, rather than merely alleviating symptoms. These results prove that the intestinal tissue regeneration effect induced by prucalopride is 5-HT4 receptor-dependent. Furthermore, it is noteworthy that such histological regenerative effects were not observed upon administration of mosapride, which is known to have the same mechanism (5-HT4 agonist) (data not shown / or described if additional comparative examples are available). This means that even though prucalopride acts on 5-HT4 receptors, it exhibits excellent and selective effects on radiation damage recovery through specific downstream signaling pathways or tissue affinity that distinguish it from other drugs.

[0071] Accordingly, the composition according to the present invention has high value as an adjuvant therapy during radiation therapy of the lower abdomen, such as the abdomen, pelvis, and rectum.

[0072] Figure 1 shows images of intestinal motility over time in mice of each experimental group. (Con: Non-irradiated control group, IR: Irradiated group, IR+Pru: Group administered prucalopride after irradiation, IR+Pru / GR: Group administered prucalopride and a 5-HT4 receptor antagonist after irradiation; GR used GR-113808, which is a drug acting as a potent and selective 5-HT4 receptor antagonist.)

[0073] Figure 2 shows images of intestinal tissue damage and the expression of intestinal epithelial markers villin and Ki-67, obtained by hematoxylin-eosin (H&E) staining and immunohistochemical staining of intestinal tissues from mice of each experimental group. (Con: Non-irradiated control group, IR: Irradiated group, IR+Pru: Group administered prucalopride after irradiation, IR+Pru / GR: Group administered prucalopride and a 5-HT4 receptor antagonist after irradiation; GR used GR-113808, which is a drug acting as a potent and selective 5-HT4 receptor antagonist.)

[0074] Figure 3 is a graph analyzing the degree of intestinal tissue damage by quantifying crypt count, villi length (um), and histological score from intestinal tissue analysis images of mice in each experimental group. (Con: Non-irradiated control group, IR: Irradiated group, IR+Pru: Group administered prucalopride after irradiation, IR+Pru / GR: Group administered prucalopride and a 5-HT4 receptor antagonist after irradiation; GR used GR-113808, which is a drug acting as a potent and selective 5-HT4 receptor antagonist.)

[0075] Figure 4 shows images of the expression of Olfm4, an intestinal stem cell marker, in mice of each experimental group. (Con: Non-irradiated control group, IR: Irradiated group, IR+Pru: Group administered prucalopride after irradiation, IR+Pru / GR: Group administered prucalopride and a 5-HT4 receptor antagonist after irradiation; GR used GR-113808, which is a drug acting as a potent and selective 5-HT4 receptor antagonist.)

[0076] Figure 5 is a graph showing the mRNA expression levels of the intestinal stem cell markers Olfm4 and Lgr5 from the intestinal tissues of mice in each experimental group. (Con: Non-irradiated control group, IR: Irradiated group, IR+Pru: Group administered prucalopride after irradiation, IR+Pru / GR: Group administered prucalopride and a 5-HT4 receptor antagonist after irradiation; GR used GR-113808, which is a drug acting as a potent and selective 5-HT4 receptor antagonist.)

[0077] Figure 6 is a graph analyzing the mRNA expression levels of the inflammatory cytokines interleukin-1 beta (IL-1β), tumor necrosis factor-alpha (TNF-α), interleukin 6 (IL-6), and monocyte chemoattractant protein-1 (MCP-1) from the intestinal tissues of mice in each experimental group. (Con: Non-irradiated control group, IR: Irradiated group, IR+Pru: Group administered prucalopride after irradiation, IR+Pru / GR: Group administered prucalopride and a 5-HT4 receptor antagonist after irradiation; GR used GR-113808, which is a drug acting as a potent and selective 5-HT4 receptor antagonist.)

[0078] Figure 7 shows images analyzing the expression of claudin 3 (Cldn3), an intestinal epithelial junction protein, by immunohistochemical staining of intestinal tissues from mice of each experimental group. (Con: Non-irradiated control group, IR: Irradiated group, IR+Pru: Group administered prucalopride after irradiation, IR+Pru / GR: Group administered prucalopride and a 5-HT4 receptor antagonist after irradiation; GR used GR-113808, which is a drug acting as a potent and selective 5-HT4 receptor antagonist.)

[0079] Figure 8 is a graph measuring the in vivo permeability of foreign substances through the intestines based on plasma FITC concentrations in mice of each experimental group. (Con: Non-irradiated control group, IR: Irradiated group, IR+Pru: Prucalopride administered after irradiation, IR+Pru / GR: Prucalopride and 5-HT4 receptor antagonist administered after irradiation; GR used GR-113808, which is a drug acting as a potent and selective 5-HT4 receptor antagonist.)

[0080] Figure 9 is a graph analyzing the mRNA expression levels of villin, zonula occludens 1 (Zo1), and claudin 3 (Cldn3), genes related to intestinal epithelial permeability, from the intestinal tissues of mice in each experimental group. (Con: Non-irradiated control group, IR: Irradiated group, IR+Pru: Prucalopride administration group after irradiation, IR+Pru / GR: Prucalopride and 5-HT4 receptor antagonist administration group after irradiation; GR used GR-113808, which is a drug acting as a potent and selective 5-HT4 receptor antagonist.)

[0081] Figure 10a is a photograph of bacterial translocation analysis using mesenteric lymph nodes.

[0082] Figure 10b is a graph of the number of bacterial translocation colonies counted.

[0083] Figure 11 shows the results confirming excellent therapeutic efficacy according to treatment with various salt forms of prucalopride.

[0084] Figure 12 shows the DSC data of prucalopride free salt.

[0085] Figure 13 shows the DSC data of prucalopride hydrochloride.

[0086] Figure 14 shows the DSC data of prucalopride oxalate.

[0087] Figure 15 shows the DSC data of prucalopride fumarate.

[0088] Figure 16 shows the DSC data of prucalopride citrate.

[0089] Figure 17 shows the DSC data of prucalopride succinate salt.

[0090] The advantages and features of the present invention and the methods for achieving them will become clear by referring to the embodiments described in detail below. However, the present invention is not limited to the embodiments disclosed below but may be implemented in various different forms. These embodiments are provided merely to ensure that the disclosure of the present invention is complete and to fully inform those skilled in the art of the scope of the invention, and the present invention is defined only by the scope of the claims.

[0091] The forms of prucalopride or various pharmaceutically acceptable salts thereof according to the present invention are not limited by the manufacturing examples and embodiments. The present invention may be applied to the prevention and treatment of intestinal injury in mammals, such as companion animals or laboratory animals undergoing radiation therapy, as well as in humans.

[0092] Preparation Example 1 Method for preparing prucalopride glass salt

[0093] The starting material 4-Amino-5-chloro-2,3-dihydro-N-(4-piperidinyl)-7-benzofurancarboxamide (2.0 g, purchased from EOS, China) was added to N,N-dimethylformamide (DMF, 120 mL) along with 1-chloro-3-methoxypropane (1.2 g, purchased from Aldrich), potassium carbonate (1.3 g), and potassium iodide (0.05 g), and stirred at 90 °C for 5 hours. After the reaction was complete, the mixture was cooled and filtered to remove inorganic salts. The filtrate was concentrated under reduced pressure, diluted with ethyl acetate (150 mL), washed with water, and dried with MgSO₄. The residue obtained after concentration under reduced pressure was recrystallized in IPA (80 mL) to obtain 1.82 g of a white crystalline solid (yield 80%). 1 H NMR (400 MHz, CD3OD) δ 7.57 (s, 1H), 4.76 (t, 2H), 3.92 - 3.78 (m, 1H), 3.41 (t,J= 6.2 Hz, 2H), 3.30 (s, 3H), 3.07 (t,J= 8.8 Hz, 2H), 2.84 (d,J= 11.8 Hz, 2H), 2.49 - 2.36 (m, 2H), 2.19 (t,J= 10.5 Hz, 2H), 1.95 (dd,J= 13.4, 3.3 Hz, 2H), 1.82 - 1.70 (m, 2H), 1.55 (qd,J=10.9, 3.6 Hz, 2H).

[0094] Using TA TGA55, DSC was analyzed by measuring the temperature from 40℃ at an N2 flow of 50ml / min to 250℃ at a heating rate of 10℃ / min, and data as shown in Fig. 12 was obtained.

[0095] Preparation Example 2 Method for preparing prucalopride hydrochloride

[0096] 1.25 g (3.41 mmol) of the prucalopride free salt of Example 1 was completely dissolved in 12.5 mL (10 V) of IPA. 0.35 g (1.0 eq, 3.41 mmol) of 35% hydrochloric acid was slowly added to the solution, followed by the addition of 12.5 mL (10 V) of acetone. The mixed solution was stirred at room temperature (25 °C) for 1 hour. The resulting white solid was recovered by filtration and vacuum dried at 40 °C to obtain 0.97 g (yield 70.6%) of a white crystalline solid. 1 H NMR (400 MHz, CD3OD) δ 7.54 (s, 1H), 4.76 (t, 2H), 4.45 (s, 1H), 4.12 - 3.98 (m, 1H), 3.51 (s, 2H), 3.46 (t,J= 5.7 Hz, 2H), 3.30 (s, 3H), 3.15 (dd,J= 16.4, 8.5 Hz, 3H), 3.04 (t,J= 8.7 Hz, 3H), 2.17 (d,J= 12.6 Hz, 2H), 1.97 (ddd,J= 15.6, 9.7, 5.8 Hz, 2H), 1.84 (s, 2H).

[0097] Using TA TGA55, DSC was analyzed by measuring with N2 flow of 50 ml / min, starting at 40℃, and increasing to 250℃ at a heating rate of 10℃ / min, and data as shown in Fig. 13 was obtained.

[0098] Preparation Example 3 Method for preparing prucalopride oxalate salt

[0099] 1.41 g (3.84 mmol) of prucalopride free salt was completely dissolved in 14.1 mL (10 V) of IPA. 0.35 g (1.0 eq, 3.84 mmol) of oxalic acid was added to the solution, and the mixture was heated and stirred under reflux (approx. 80 °C) for 1 hour. The reaction mixture was cooled to room temperature and stirred for an additional 1 hour. The resulting white solid was recovered by filtration and vacuum dried at 50 °C to obtain 1.70 g of a white crystalline solid (yield 96.8%).1 H NMR (400 MHz, CD3OD) δ 7.55 (s, 1H), 4.76 (t, 2H), 4.07 (s, 1H), 3.61 (s, 2H), 3.45 (t,J= 5.7 Hz, 2H), 3.31 (s, 3H), 3.19 (s, 2H), 3.05 (t,J= 8.7 Hz, 4H), 2.18 (d,J= 12.2 Hz, 2H), 1.97 (dt,J= 15.7, 5.8 Hz, 2H), 1.83 (s, 2H).

[0100] Using TA TGA55, DSC was analyzed by measuring the temperature from 40℃ at an N2 flow of 50ml / min to 250℃ at a heating rate of 10℃ / min, and data as shown in Fig. 14 was obtained.

[0101] Preparation Example 4 Method for preparing prucalopride fumarate

[0102] 1.70 g (4.63 mmol) of prucalopride free salt was completely dissolved in 17 mL (10 V) of IPA. 0.54 g (1.0 eq, 4.63 mmol) of fumaric acid was added to the solution, and the mixture was stirred for 1 hour under reflux (approx. 80 °C). The reaction mixture was cooled to room temperature (25 °C) and stirred for an additional 1 hour. The resulting white solid was recovered by filtration and vacuum dried at 45–50 °C to obtain 1.76 g of a white crystalline solid (yield 78.65%). 1H NMR (400 MHz, CD3OD) δ 7.55 (s, 1H), 6.66 (s, 2H), 4.76 (t,J= 8.8 Hz, 2H), 4.14 - 3.98 (m, 1H), 3.50 (d,J= 12.3 Hz, 2H), 3.45 (t,J= 5.8 Hz, 2H), 3.31 (s, 3H), 3.13 (dd,J= 8.8, 7.0 Hz, 2H), 3.04 (dd,J= 15.1, 6.4 Hz, 4H), 2.16 (dd,J= 14.1, 2.8 Hz, 2H), 1.96 (ddd,J= 15.9, 9.7, 5.8 Hz, 2H), 1.83 (td,J= 14.5, 3.8 Hz, 2H).

[0103] Using TA TGA55, DSC was analyzed by measuring the starting temperature at 40℃ and increasing the temperature at a rate of 10℃ / min up to 250℃ with an N2 flow of 50ml / min, and data as shown in Fig. 15 was obtained.

[0104] Preparation Example 5 Method for preparing prucalopride citrate

[0105] 1.79 g (4.88 mmol) of prucalopride free salt was completely dissolved in 17.9 mL (10 V) of IPA. 1.03 g (1.0 eq, 4.88 mmol) of citric acid was added to the solution, and the mixture was stirred under reflux (approx. 80 °C) for 1 hour. Afterward, the reaction mixture was cooled to room temperature and stirred for an additional 1 hour.

[0106] The formed solid was filtered and recovered, and dried at 45°C under vacuum to obtain 2.32g of white crystalline solid (yield 85%). 1H NMR (400 MHz, CD3OD) δ 7.54 (s, 1H), 4.73 (t,J= 8.8 Hz, 2H), 4.14 - 3.98 (m, 1H), 3.50 (d,J= 12.5 Hz, 2H), 3.45 (t,J= 5.8 Hz, 2H), 3.30 (s, 3H), 3.18 - 3.09 (m, 2H), 3.04 (dd,J= 15.6, 6.9 Hz, 4H), 2.74 (dd,J= 36.1, 15.4 Hz, 4H), 2.23 - 2.10 (m, 2H), 2.01 - 1.93 (m, 2H), 1.87 (d,J= 11.7 Hz, 2H).

[0107] Using TA TGA55, DSC was analyzed by measuring the temperature from 40℃ at an N2 flow of 50ml / min to 250℃ at a heating rate of 10℃ / min, and data as shown in Fig. 16 was obtained.

[0108] Preparation Example 6 Method for preparing prucalopride succinate salt

[0109] 10 g (0.028 mol) of prucalopride free salt was added to 100 mL of IPA and completely dissolved. 3.4 g (0.029 mol) of succinic acid was added to the solution, and the mixture was stirred at 45 °C for 30 minutes. Afterward, the reaction solution was cooled to room temperature and stirred for an additional hour. The formed solid was recovered by filtration and dried under vacuum at 45 °C to obtain 10.89 g of a white crystalline solid (yield 80%). 1H NMR (400 MHz, DMSO) δ 7.42 (s, 1H), 7.23-7.25 (d,J= 7.2 Hz, 1H), 5.83 (s, 2H), 4.70-4.66 (t,J= 8.8 Hz, 2H), 3.77-3.69 (m, 1H), 3.28-3.31 (t,J= 6.4 Hz, 2H), 3.17 (s, 3H), 2.97-3.01 (t,J= 8.8 Hz, 2H), 2.75-2.78 (m, 2H), 2.38-2.39 (m, 2H), 2.35 (s, 4H), 2.14-2.19 (t,J= 10.3 Hz, 2H), 1.78-1.82 (m, 2H), 1.60-1.66 (m, 2H), 1.40-1.48 (m, 2H).

[0110] Using TA TGA55, DSC was analyzed by measuring the starting temperature at 40℃ and increasing the temperature at a rate of 10℃ / min up to 250℃ with an N2 flow of 50ml / min, and data as shown in Fig. 17 was obtained.

[0111] Experimental Example 1: Assignment of Experimental Animals

[0112] The mice used in this experiment were 6-week-old male C57BL / 6 mice purchased from Orient Bio (Seongnam, Gyeonggi-do, South Korea) and acclimatized for one week in a Specific Pathogen-Free (SPF) facility before being used in the experiment. The mice were randomly assigned to one of the following groups: a non-irradiated control group (Con), an irradiated group (IR), a group administered prucalopride after irradiation (IR+Pru), and a group administered prucalopride and a 5-HT4 receptor antagonist after irradiation.

[0113] Specifically, the above GR is GR-113808, and is a drug that acts as a potent and selective 5-HT4 receptor antagonist.

[0114] Experimental Example 2: Radiation Irradiation Method

[0115] Prior to the experiment, the mice were anesthetized, and then 13.5 Gy of radiation was irradiated to the lower abdominal area including the intestines using an X-RAD 320 X-ray irradiator (Precision X-Ray, East Haven, CT, USA; filter: 2 mm AI, 42 cm, 260 kV / s, 10 mA, 2.0 Gy / min).

[0116] After irradiation, mice were orally administered 5 mg / kg of prucalopride (Resolor®, Janssen) once daily for 6 days, and some of the mice that received oral prucalopride were intraperitoneally administered 5 mg / kg of GR-113808 (Sigma), a drug acting as a potent and selective 5-HT4 receptor antagonist, once daily. On the 6th day after irradiation, the mice were euthanized, and intestines were collected for histological analysis.

[0117] Experimental Example 3: Intestinal Motility Analysis

[0118] Barium sulfate contrast agent was orally administered to mice in each experimental group, and intestinal motility was analyzed through X-ray imaging.

[0119] Experimental Example 4: Analysis of the Degree of Histological Damage Recovery

[0120] The degree of intestinal tissue damage was measured by hematoxylin-eosin (H&E) staining and immunohistochemical staining of the intestinal tissue of each experimental group of mice according to Example 2, the expression of the intestinal epithelial marker villin and the cell proliferation marker Ki-67 was measured, and the degree of intestinal tissue damage and the degree of recovery of intestinal tissue damage after radiation were analyzed by quantifying the Crypt count, Villi length (um), and Histological score.

[0121] Experimental Example 5: Analysis of Intestinal Stem Cell Recovery Degree

[0122] The degree of intestinal tissue damage in mice of each experimental group was measured, and Olfm4 expression was indicated to quantify and analyze the mRNA expression levels of Olfm4 and Lgr5 from the intestinal tissue.

[0123] Experimental Example 6: Analysis of Anti-inflammatory Degree

[0124] The degree of infiltration of inflammatory cells in the tissue was measured through immunohistochemical staining of CD68, an inflammatory cell marker, and the mRNA gene expression of inflammatory factors IL-1β, TNF-α, IL-6, and MCP-1 was quantified using qPCR analysis.

[0125] Experimental Example 7: Analysis of Barrier Function {Fluorescein isothiocyanate (FITC)-dextran absorption assay and bacterial translocation assay}

[0126] The permeability of the mouse intestinal barrier was evaluated using the FITC-dextran absorption assay. On the third day after irradiation, mice were anesthetized, the abdominal cavity was opened, and a 5 cm segment of the distal ileum was blocked using a clamp. Then, 12.5 mg of FITC-dextran (4 kDa, Sigma, St Louis, MO) diluted in 100 µl of PBS was injected into the ileum. Thirty minutes after injection, blood was collected via cardiac puncture, and the concentration of FITC in the serum samples was analyzed using a fluorescence spectrophotometer (filtration, 485 nm; absorption, 528 nm).

[0127] Mesenteric lymph nodes were collected from mice in each experimental group, and tissue fragments were cultured on McConkey agar to analyze bacterial translocation to the mesenteric lymph nodes.

[0128] The expression of Villin and Cldn3 proteins, which constitute cell-to-cell junctions, was stained using immunohistochemical staining in the intestinal tissues of mice from each experimental group collected on the 6th day after irradiation, and the mRNA expression of Villin, Zo1, and Cldn3 was quantified through Real-Time RT-PCR.

[0129] Example 1: Confirmation of the intestinal motility-promoting effect of prucalopride in a mouse model with radiation-induced intestinal injury

[0130] Intestinal motility was analyzed as described in Experimental Example 3, and the results are shown in Fig. 1.

[0131] According to Figure 1, a decrease in motility was observed in the irradiated mice, whereas it was confirmed that the decrease in motility was improved in the prucalopride-treated mice (IR+Pru) after irradiation. In the prucalopride and GR-113808 (5-HT4 receptor antagonist) administration group (IR+Pru / GR) after irradiation, it was confirmed that intestinal motility decreased similarly to that of the irradiated mice (IR).

[0132] Example 2: Confirmation of the intestinal tissue regenerative effect of prucalopride in a mouse model with radiation-induced intestinal damage

[0133] Mouse intestinal tissue collected by the method of Experimental Example 2 was fixed with a 10% neutral buffered formalin solution, paraffin blocks were prepared, and sections were made to a thickness of 4 µm. Subsequently, the intestinal sections were stained with hematoxylin-eosin (H&E).

[0134] The analysis of histological changes in intestinal tissue damage in the ileum region was evaluated based on the degree of epithelial damage (0–3 points), the degree of crypt destruction (0–3 points), the degree of microvascular dilation (0–3 points), and the degree of mucosal inflammatory response (0–3 points), and the scores of each item were summed.

[0135] In addition, the regenerative capacity and degree of damage of the crypt were confirmed through immunohistochemical staining of Ki-67, a cell proliferation marker.

[0136] Histopathological analysis was performed on mouse small intestine tissues collected on the 6th day after abdominal irradiation with 13.5 Gy, as described in Experimental Examples 2 and 4. As a result, in the irradiated mice (IR), crypt destruction, villi shortening, loss of the epithelial layer, infiltration of inflammatory cells in the mucosa and submucosa, and epithelial proliferation disorders (confirmed via marker Ki-67) were observed. This is shown in Figure 2.

[0137] In comparison, in mice treated with prucalopride after irradiation (IR+Pru), the length of villi and the number of crypts in the small intestine were statistically significantly higher than in mice treated with irradiation (IR), and the total score for the degree of epithelial damage (0~3 points), degree of crypt destruction (0~3 points), degree of microvascular dilation (0~3 points), and degree of mucosal inflammatory response (0~3 points) was found to be significantly lower than in the irradiation group (IR).

[0138] Meanwhile, in the group administered prucalopride and GR-113808 (a 5-HT4 receptor antagonist) after radiation (IR+Pru / GR), crypt destruction, villi shortening, loss of the epithelial layer, infiltration of inflammatory cells in the mucosa and submucosa, and impaired epithelial proliferation (confirmed via marker Ki-67) were observed, similar to the radiation-treated mice (IR), confirming that prucalopride acts as a therapeutic agent for intestinal tissue damage. This is illustrated in Figure 3.

[0139] Through the above results, it was confirmed that prucalopride has a significant regenerative effect on intestinal tissue damaged by radiation.

[0140] Example 3: Confirmation of the intestinal stem cell recovery effect of prucalopride in a mouse model with radiation-induced intestinal damage

[0141] The effect of procalopride on intestinal stem cells of irradiated mice was analyzed according to the method described in Experimental Example 5.

[0142] The degree of damage to the intestinal tissue of each experimental group of mice was measured, and the expression of Olfm4 was observed, as shown in Figure 4.

[0143] The mRNA expression levels of Olfm4 and Lgr5 were quantified and analyzed, and higher mRNA expression was confirmed in mice treated with procalopride after irradiation (IR+Pru) compared to mice irradiated with radiation (IR).

[0144] Meanwhile, in mice (IR+Pru / GR) administered prucalopride and GR-113808 (a 5-HT4 receptor antagonist) after radiation, low mRNA expression was confirmed, confirming that prucalopride acts as a therapeutic agent for intestinal tissue damage. This is shown in Figure 5.

[0145] Through the above results, it was confirmed that prucalopride has a significant effect in restoring intestinal stem cells damaged by radiation.

[0146] Example 4: Confirmation of the anti-inflammatory effect of prucalopride in a mouse model with radiation-induced intestinal injury

[0147] The effect of prucalopride on intestinal inflammation in irradiated mice was analyzed according to the method described in Experimental Example 6.

[0148] The level of CD68, a macrophage marker, indicates the degree of inflammation in intestinal diseases. A significantly higher number of CD68-positive cells was observed in the intestinal tissue of irradiated mice (IR) compared to non-irradiated control mice (Con). Additionally, it was confirmed that the number of macrophages increased by radiation was significantly suppressed in prucalopride-treated mice (IR+Pru) after irradiation. This is shown in Figure 6.

[0149] In addition, compared to the non-irradiated normal control mice (Con), the mRNA levels of the inflammatory markers IL-1β, TNF-α, IL-6, and MCP-1 in the intestinal tissue of the irradiated mice (IR) were significantly increased, whereas they were significantly decreased in the prucalopride-treated mice (IR+Pru) after irradiation.

[0150] Meanwhile, in the group administered prucalopride and GR-113808 (a 5-HT4 receptor antagonist) after radiation (IR+Pru / GR), mRNA levels of inflammatory markers IL-1β, TNF-α, IL-6, and MCP-1 significantly increased, similar to the irradiated mice (IR), confirming that prucalopride acts as a treatment for intestinal inflammation. This is shown in Figure 6.

[0151] Through the above results, it was confirmed that prucalopride has a significant effect in alleviating the inflammatory response of the intestines induced by radiation.

[0152] Example 5: Confirmation of the barrier function restoration effect of prucalopride in a mouse model with radiation-induced intestinal damage

[0153] The effect of prucalopride on the small intestinal epithelial barrier function of irradiated mice was analyzed according to the method described in Experimental Example 7.

[0154] First, the permeability of the mouse intestinal barrier was evaluated through FITC-dextran absorption analysis. In the irradiated mice (IR), serum FITC concentration increased, while in the prucalopride-treated mice (IR+Pru), it was found to decrease.

[0155] Meanwhile, it was confirmed that serum FITC concentrations increased in the groups administered prucalopride and GR-113808 (a 5-HT4 receptor antagonist) after radiation (IR+Pru / GR), similar to the radiation-treated mice (IR), as shown in Figure 8.

[0156] In addition, the expression of epithelial junction molecules was investigated using immunohistochemical staining of collected intestinal tissue. It was confirmed that the expression of villin, Zo1, and Cldn3, which are constituent proteins of epithelial junction that are reduced by radiation, increased in prucalopride-treated mice (IR+Pru) after irradiation. The results of the immunohistochemical staining are shown in Figure 7, and the results of comparing the relative mRNA amounts of villin, Zo1, and Cldn3, which are constituent proteins of epithelial junction, are shown in Figure 9.

[0157] Meanwhile, in the group administered prucalopride and GR-113808 (a 5-HT4 receptor antagonist) after radiation (IR+Pru / GR), it was confirmed that the expression of villin, Zo1, and Cldn3 decreased similarly to that of the radiation-treated mice (IR), as shown in Figure 9.

[0158] An analysis of lymph node colony counts was performed using mesenteric lymph nodes to analyze bacterial migration, and the results are shown in Fig. 10a. The results of Fig. 10a were diagrammed and are shown in Fig. 10b.

[0159] According to Figures 10a and 10b, it was confirmed that the increase in the number of colonies due to bacterial migration to mesenteric lymph nodes in the irradiated mice (IR) was significantly reduced in the prucalopride-treated mice (IR+Pru) after irradiation.

[0160] Meanwhile, in the group administered prucalopride and GR-113808 (a 5-HT4 receptor antagonist) after radiation (IR+Pru / GR), the number of colonies decreased similarly to that of the irradiated mice (IR), confirming that prucalopride acts as a treatment for intestinal inflammation. This is shown in Figure 10.

[0161] Through the above results, it was confirmed that prucalopride acts as a treatment for radiation-induced intestinal inflammation and has a significant restorative effect on intestinal epithelial barrier function.

[0162] Example 6: Confirmation of therapeutic efficacy according to the salt type of prucalopride

[0163] Similar to the method of Experimental Example 2, the therapeutic efficacy against radiation-induced intestinal inflammation was confirmed using hematoxylin-eosin (H&E) while varying the salt types of prucalopride. Specifically, succinate, fumarate, hydrochloride, citrate, oxalate, and free salts were obtained and administered to an animal model similar to the method of Experimental Example 2. Mouse intestinal tissue collected after administration was fixed with a 10% neutral buffered formalin solution, paraffin blocks were prepared, and the tissue was sectioned to a thickness of 4 µm. Subsequently, the intestinal sections were stained with hematoxylin-eosin (H&E) to confirm the therapeutic efficacy, and the results are shown in Fig. 11. The analysis of histological changes in intestinal tissue damage in the ileum region was evaluated based on the degree of epithelial damage (0–3 points), the degree of crypt destruction (0–3 points), the degree of microvascular dilation (0–3 points), and the degree of mucosal inflammatory response (0–3 points), and the scores of each item were summed.

[0164] As can be seen in Fig. 11, all of the prucalopride succinate, fumarate, hydrochloride, citrate, oxalate, and free salts used exhibited therapeutic efficacy. This demonstrates that the pharmacological effect of prucalopride is not limited to specific salts but is due to its basic skeletal structure itself.

[0165] In particular, among the various salt forms, it was confirmed that prucalopride free salt, succinate, and citrate were highly effective.

Claims

1. A pharmaceutical composition for the prevention or treatment of intestinal damage caused by radiation treatment, comprising prucalopride or a pharmaceutically acceptable salt, hydrate, or co-crystal thereof as an active ingredient.

2. A pharmaceutical composition according to claim 1, wherein prucalopride is a compound represented by the following chemical formula 1: [Chemical Formula 1] 3. A pharmaceutical composition according to claim 1, characterized in that the pharmaceutically acceptable salt is selected from the group consisting of succinate, citrate, fumarate, hydrochloride, and oxalate.

4. A pharmaceutical composition according to claim 1, wherein the radiation treatment is by radiation therapy.

5. A pharmaceutical composition according to claim 1, wherein the intestinal injury is one or more selected from the group consisting of inflammation, edema, tumor, ulcer, scar, and stenosis.

6. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition promotes the expression of one or more genes selected from the group consisting of Villin, Olfm4 (olfactomedin-4), Lgr5, Cldn3 (claudin-3), and Zo-1.

7. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition inhibits changes in gene expression of one or more selected from the group consisting of IL-1β, TNF-α, IL-6, and Mcp-1.

8. A food composition for preventing or improving intestinal damage caused by radiation treatment, comprising prucalopride or its food-grade acceptable salts, hydrates, or co-crystals as an active ingredient.

9. In claim 8, a food composition wherein prucalopride is a compound represented by the following chemical formula 1: [Chemical Formula 1] 10. A food composition according to claim 8, wherein the radiation treatment is by radiation therapy.

11. A food composition according to claim 8, wherein the intestinal damage is one or more selected from the group consisting of inflammation, edema, tumor, ulcer, scar, and stenosis.

12. The food composition of claim 8, wherein the food composition promotes the expression of one or more genes selected from the group consisting of Villin, Olfm4 (olfactomedin-4), Lgr5, Cldn3 (claudin-3), and Zo-1.

13. The food composition of claim 8, wherein the food composition inhibits changes in gene expression of one or more selected from the group consisting of IL-1β, TNF-α, IL-6, and Mcp-1.

14. A method for treating intestinal damage caused by radiation treatment, comprising the step of administering prucalopride or a pharmaceutically acceptable salt, hydrate, or co-crystal thereof to a subject requiring it.

15. A method according to paragraph 14, wherein the radiation treatment is by radiation therapy.

16. A method according to claim 14, wherein the intestinal injury is one or more selected from the group consisting of inflammation, edema, tumor, ulcer, scar, and stenosis.

17. Use of the prucalopride of the present invention or a pharmaceutically acceptable salt thereof for manufacturing a medicine for the prevention or treatment of intestinal damage caused by radiation treatment.

18. In paragraph 17, the above radiation treatment is by radiation therapy.

19. In paragraph 17, the above-mentioned intestinal injury is one or more selected from the group consisting of inflammation, edema, tumor, ulcer, scar, and stenosis.

20. A composition comprising prucalopride or a pharmaceutically acceptable salt thereof for use in the prevention or treatment of intestinal damage caused by radiation treatment.

21. A composition according to claim 20, wherein the intestinal injury is one or more selected from the group consisting of inflammation, edema, tumor, ulcer, scar, and stenosis.

22. The method according to claim 14, characterized in that the prucalopride or its pharmaceutically acceptable salt, hydrate, solvate, or cocrystal is administered at a dose of 0.1 mg to 100 mg per day for adults.

Images