Composition comprising niclosamide or pharmaceutically acceptable salt thereof for preventing formation of pathological extracellular matrix (ECM) or alleviating or improving preformed pathological extracellular matrix, and administration method using same

Abstract

The present invention relates to: a composition for preventing the formation of a pathological extracellular matrix (ECM) or alleviating or improving a preformed pathological extracellular matrix, the composition comprising niclosamide or a pharmaceutically acceptable salt thereof; and an administration method using same.

Description

A composition for preventing the formation of pathological extracellular matrix (ECM) comprising niclosamide or a pharmaceutically acceptable salt thereof, or for alleviating or improving a formed pathological extracellular matrix, or a method of administration using the same.

[0001] The present invention relates to a composition for the prevention, alleviation, or improvement of a pathological extracellular matrix containing niclosamide, and a method of administration using the same.

[0002]

[0003] In general, the Extracellular Matrix (ECM) not only functions as a structural support for tissues but also plays a crucial role in maintaining tissue homeostasis by regulating cell adhesion, migration, differentiation, proliferation, and signal transduction. Under normal conditions, the ECM is dynamically regulated through a balance of production and degradation; however, if this balance is disrupted by the progression of specific diseases or external stimuli, abnormal changes in the structure and composition of the ECM may occur. Such pathological ECM can be induced by two main mechanisms.

[0004] First, this refers to cases where the generation and reorganization of the ECM are promoted by various factors (e.g., growth factors, cytokines, chemokines, etc.) secreted by cells or stromal cells as the disease progresses. For instance, in conditions such as cancer, neurodegenerative diseases, diabetes, chronic inflammatory diseases, and metabolic diseases, factors secreted from cells or surrounding stromal cells during the progression of the disease can induce the excessive accumulation or abnormal arrangement of ECM components, such as collagen, proteoglycans, and fibronectin, leading to ECM rigidification and a decline in tissue function.

[0005] Second, there are cases where the structure and composition of the ECM are altered or tissue hardening is induced by the long-term and repeated administration of external drugs. Various drugs, such as anticancer agents, immunotherapies, targeted therapies, antibody therapies, radiation therapy, and hormone therapy, as well as treatments for neurological diseases, anti-inflammatory agents, and immunomodulators, exert a continuous influence on the tissue microenvironment when administered over a long period or repeatedly. As a result, the remodeling or re-hardening of the ECM may be induced. These ECM modifications induced by external drugs can gradually accumulate during the process of repeated drug administration, leading to tissue rigidity, ECM overaccumulation, and reduced drug penetration.

[0006] Particularly in patients undergoing continuous repetitive chemotherapy, pathological extracellular matrix (ECM) can form not only within the tumor tissue but also in organs susceptible to metastasis. This can hinder drug delivery within the tumor and cause the tumor microenvironment to transition into a treatment-resistant state. Furthermore, excessive accumulation or alteration of the ECM can impede drug penetration into the tumor tissue, leading to a gradual shift of the tumor microenvironment toward a treatment-resistant state. It has also been suggested that the modified ECM may accelerate the progression of metastatic cancer by forming a pre-metastatic niche prior to metastasis, thereby providing an environment favorable for the colonization of circulating tumor cells (CTCs). This phenomenon is distinct from conventional resistance caused by changes in drug sensitivity due to genetic modifications, and can be understood as a form of treatment resistance resulting from changes in the tissue environment itself during repeated treatments or drug administration. Furthermore, pathological ECM is not limited to this but may similarly occur in degenerative brain diseases (such as Parkinson's disease and Alzheimer's disease) where repeated drug administration takes place, as well as in diabetes, chronic inflammatory diseases, and metabolic diseases. In addition, abnormal accumulation and rigidification of the ECM can act as important factors in the pathophysiology of diseases accompanied by the strengthening of the ECM due to specific stimuli or treatments, such as benign prostatic hyperplasia (BPH), thyroid nodules, polycystic ovary syndrome (PCOS), radiation-induced tissue sclerosis (radiation fibrosis), and lymphedema.For example, in benign prostatic hyperplasia, repetitive stimulation by male hormones (androgens) can lead to the excessive accumulation of ECM components, such as collagen and proteoglycans, within the prostate; in polycystic ovary syndrome, persistent endocrine imbalances, such as excess androgens and insulin resistance, can result in an increase in ECM components (particularly collagen) in the ovarian cortex; and radiation-induced tissue sclerosis can manifest as long-term side effects in which the tissue gradually hardens as the ECM is abnormally reorganized due to repeated radiation exposure. In the case of lymphedema, continuous lymphatic fluid retention and compression following lymphatic vessel damage or lymph node removal can induce ECM remodeling, leading to tissue rigidity.

[0007] Meanwhile, chemotherapy is a widely used treatment method to suppress or destroy rapidly proliferating cancer cells, but it affects tissues that divide rapidly among normal cells (e.g., bone marrow, digestive mucosa, hair follicles, etc.) and causes various side effects. Representative examples include leukopenia and increased risk of infection due to bone marrow suppression, anemia and fatigue, and increased risk of bleeding due to thrombocytopenia. Additionally, gastrointestinal disorders (nausea, vomiting, diarrhea, constipation), oral mucositis, hair loss, neurotoxicity (peripheral neuropathy, cognitive decline), cardiotoxicity, nephrotoxicity, immunosuppression, and increased risk of thrombosis have been reported. To alleviate these side effects, antiemetics (e.g., 5-HT3 antagonists), hematopoietic growth factors (e.g., G-CSF), anti-inflammatory drugs (NSAIDs), immunosuppressants such as steroids, and antithrombotic agents (e.g., warfarin, heparin) are used, but there are other limitations with long-term use, such as constipation, headache, bone pain, increased risk of thrombosis, gastrointestinal disorders and kidney damage, increased risk of infection and osteoporosis, risk of bleeding and complex drug interactions.

[0008] Furthermore, Acquired Drug-Induced Fibrosis (ADIF) is raised as one of the side effects caused by long-term drug administration. While general pathological fibrosis occurs due to tissue repair processes following necrosis or apoptosis and chronic inflammation, often accompanied by irreversible damage, ADIF is characterized by tissue stiffening induced by abnormal ECM remodeling mediated by mitochondrial damage, increased metabolic stress, and increased reactive oxygen species (ROS) production following repeated drug administration; it is suggested that ADIF may exhibit some reversible aspects. However, in actual clinical practice or treatment, discontinuing drug administration to alleviate side effects can lead to a decrease in anticancer or disease treatment efficacy; therefore, a method is required to effectively control side effects while continuing treatment.

[0009] Although therapeutic strategies such as combination therapy or cross-administration have recently been attempted, these methods have limitations in that they cannot directly regulate the pathological ECM itself. Furthermore, the continuous administration of drugs with identical or similar toxic mechanisms may actually accelerate side effects such as mucositis, nausea and vomiting, neurological side effects, and leukopenia. Therefore, there is a need to develop novel pharmaceutical compositions and administration methods that can maintain or enhance drug delivery efficiency and therapeutic efficacy, and ensure long-term therapeutic sustainability, by effectively preventing, mitigating, or improving the pathological ECM—which encompasses both ECM modifications induced by factors secreted from cells or stromal cells during disease progression and tissue sclerosis induced by the repeated administration of external drugs.

[0010]

[0011] The present invention aims to solve the problem of reduced tissue delivery efficiency and decreased therapeutic efficacy caused by pathological extracellular matrix (ECM) that develops during repetitive or long-term treatment processes. Specifically, the present invention utilizes a composition containing niclosamide or its pharmaceutically acceptable salts and / or hydrates as active ingredients to address the problem of abnormal deformation and accumulation of the ECM induced by factors such as growth factors, cytokines, and chemokines secreted from cells or stromal cells as the disease progresses. Furthermore, the present invention aims to effectively prevent, alleviate, or improve pathological ECM formed by at least one of tissue hardening and ECM re-strengthening induced by the long-term and repetitive administration of external drugs, such as anticancer agents, treatments for neurological diseases, anti-inflammatory agents, and immunomodulators, by utilizing a composition containing niclosamide or its pharmaceutically acceptable salts and / or hydrates as active ingredients.

[0012] Furthermore, the present invention aims to address the problem that such pathological ECM commonly appears in various diseases such as cancer, neurodegenerative diseases, diabetes, chronic inflammatory diseases, and metabolic diseases, and that the formation of pathological ECM reduces the penetration and absorption rates of drugs into tissues, resulting in a gradual decrease in therapeutic efficacy despite repeated drug administration.

[0013] Furthermore, the present invention aims to address the problem that during repetitive anticancer treatment, pathological ECM converts the tumor microenvironment into a treatment-resistant state or forms a pre-metastatic niche, thereby promoting the engraftment of metastatic cancer cells. At the same time, the invention aims to provide a novel approach capable of controlling ECM rigidization phenomena—such as Acquired Drug-Induced Fibrosis (ADIF), which are caused by repeated drug administration but are relatively reversible—without compromising therapeutic efficacy.

[0014] Furthermore, the present invention aims to overcome the limitations of existing combination or cross-administration methods—namely, the problem of failing to improve ECM rigidity itself and merely accumulating side effects—by providing a composition and administration strategy capable of directly regulating the pathological ECM itself. In particular, a key objective of the present invention is to simultaneously ensure the continuity and efficiency of treatment in diseases requiring long-term therapy by suppressing or improving the pathological ECM without interrupting treatment.

[0015]

[0016] To solve the aforementioned problem, the present invention provides a composition for preventing the formation of a pathological extracellular matrix (ECM) or for alleviating or improving a formed pathological extracellular matrix, comprising niclosamide or a pharmaceutically acceptable salt thereof as an active ingredient.

[0017] It has been disclosed that niclosamide has effects that inhibit fibrosis or induce apoptosis. However, to achieve the effects of inhibiting fibrosis or inducing apoptosis, niclosamide had to be administered at high doses, and niclosamide exhibited cytotoxicity when administered at high doses, which has been a limitation that has made it practically difficult to commercialize. However, the present invention was devised by confirming that a completely new therapeutic effect can be induced when niclosamide or a pharmaceutically acceptable salt thereof is continuously administered at a specific dose.

[0018] In addition, the present invention provides a method of administration for preventing the formation of pathological extracellular matrix (ECM) or alleviating or improving a formed pathological extracellular matrix, comprising the step of orally administering a composition containing niclosamide or a pharmaceutically acceptable salt thereof as an active ingredient.

[0019] In addition, the present invention provides a method for co-administration to prevent the formation of pathological extracellular matrix (ECM) or to alleviate or improve a formed pathological extracellular matrix, comprising: a pretreatment step of orally administering a composition containing niclosamide or a pharmaceutically acceptable salt thereof as an active ingredient for 3 to 7 days prior to drug administration; and a step of administering the drug in combination with the composition after the pretreatment step.

[0020] In addition, the present invention provides a method of co-administration for preventing the formation of pathological extracellular matrix (ECM) or alleviating or improving a formed pathological extracellular matrix, comprising the step of co-administering a drug with a composition containing niclosamide or a pharmaceutically acceptable salt thereof as an active ingredient.

[0021]

[0022] According to the present invention, a composition comprising niclosamide or a pharmaceutically acceptable salt thereof as an active ingredient or a method of administration using the same can effectively prevent, alleviate, or improve pathological extracellular matrix (ECM) formed by the progression of a disease or repeated administration of external drugs.

[0023] Specifically, the composition according to the present invention can effectively suppress or improve the excessive accumulation of ECM, abnormal reorganization, and tissue rigidity by directly regulating the pathological ECM formed by at least one of ECM modification induced by factors secreted from cells or stromal cells as the disease progresses and tissue hardening induced by repeated administration of external drugs. More specifically, tissue hardening induced by repeated administration of external drugs may refer to acquired drug-induced fibrosis (ADIF).

[0024] Furthermore, by reducing the spatial barrier within the tissue through the alleviation or improvement of the pathological ECM via the composition and / or administration method according to the present invention, the efficiency of tissue penetration and intracellular delivery of therapeutic drugs, such as anticancer agents, treatments for neurological diseases, anti-inflammatory agents, and immunomodulators, is enhanced. Consequently, the decrease in drug absorption rates is mitigated even during repeated drug administration, and the continuous maintenance or recovery of therapeutic efficacy becomes possible.

[0025] In addition, by inhibiting treatment-resistant changes in the tumor microenvironment and the formation of metastatic niches induced by the pathological ECM during repeated anticancer treatment processes using the composition and / or administration method according to the present invention, the effect of reducing the possibility of circulating cancer cell colonization and lowering the risk of progression of metastatic cancer is provided.

[0026] In addition, by reducing the expression level of one or more of MMP-9 and VEGF factors using the composition and / or administration method according to the present invention, the imbalance between ECM degradation and remodeling is alleviated, and pathological ECM remodeling is normalized. Through this, ECM rigidity and tissue structural abnormalities can be improved more fundamentally.

[0027] In addition, the present invention allows for the stable maintenance of the pathological ECM improvement effect while minimizing the risk of toxicity and side effects that may occur during long-term administration by administering a composition containing niclosamide or a pharmaceutically acceptable salt thereof as an active ingredient multiple times orally.

[0028] In addition, the pretreatment and combination administration method according to the present invention provides the effect of maintaining or enhancing the efficacy of existing treatments by administering a therapeutic drug after improving the ECM environment before the pathological ECM is reinforced or in a state where it has already been formed.

[0029] In addition, the composition and / or administration method of the present invention is applicable to various diseases involving pathological ECM, such as cancer, neurodegenerative diseases, diabetes, chronic inflammatory diseases, and metabolic diseases, and can be usefully utilized not only in anticancer treatment but also in various treatment situations requiring long-term and repeated drug administration.

[0030]

[0031] Figure 1 is a diagram of the experimental schedule for the administration method of the composition (CP-EFR) of Example 2 and polytaxel for the patient.

[0032] Figure 2 shows the results of administration to patient A.

[0033] Figure 3 shows the results of administration to patient B.

[0034] Figure 4 is a graph showing the degree of signal expression for each group regarding MMP9 signals related to ECM remodeling.

[0035] Figure 5 is a graph showing the degree of signal expression for each group regarding VEGF signals related to ECM remodeling.

[0036] Figure 6 shows the changes in tumor size after treatment for each group.

[0037] Figure 7 shows the MPP of Niclosamide + This is a graph confirming the protective effect against induced SH-SY5Y cytotoxicity.

[0038] Figure 8 is a graph showing the change in tumor size of the untreated group (vehicle), the paclitaxel monotherapy group, and the combination of the composition of Example 2 (CP-EFR) and paclitaxel.

[0039] Figure 9 is a comparative photograph of changes in tumor size after administration to groups G1 to G3 in Experimental Example 4-2. Specifically, in the group administered with Paclitaxel and CP-EFR, severe lung metastasis, which is observed in the group administered with Paclitaxel alone, was not observed. In the group administered with Paclitaxel alone, a rapid increase in lung metastasis was confirmed compared to the vehicle control group. On the other hand, in the group administered with Paclitaxel and CP-EFR, it was confirmed that no metastasis occurred because circulating tumor cells (CTCs) could not settle in the organ (lungs).

[0040] Figure 10 is a graph showing the change in body weight of mice administered an anticancer drug alone and an anticancer drug in combination with niclosamide in Experimental Example 5.

[0041] Figure 11 is a graph showing the rate of change in body weight in groups G1 to G4 for each mouse in Experimental Example 5.

[0042] Figure 12 is a graph showing the change in body weight of mice administered an anticancer drug alone and an anticancer drug in combination with niclosamide in Experimental Example 6.

[0043] Figure 13 is a graph showing the rate of change in body weight in the G1 and G2 groups for each mouse in Experimental Example 6.

[0044] The present invention aims to prevent the formation of pathological extracellular matrix (ECM) or to provide a composition comprising niclosamide or a pharmaceutically acceptable salt thereof as an active ingredient. It has been confirmed that when a composition comprising niclosamide or a pharmaceutically acceptable salt thereof according to the present invention is administered repeatedly and continuously, it can improve the tissue penetration rate of drugs reduced by ECM reinforcement and overcome treatment resistance by inhibiting ECM modification. Through the action of niclosamide, the overexpression of ECM constituent proteins (collagen I, III, VI, fibronectin, etc.) is inhibited, and the ECM is maintained in a normal state, thereby enabling the effective delivery of anticancer drugs and other therapeutic agents into the tissue. Furthermore, by forming a drug penetration pathway through ECM remodeling and maximizing therapeutic responsiveness, a sustained therapeutic effect can be maintained even during repeated anticancer treatments.

[0045] In this specification, the term "pathological extracellular matrix (ECM)" may refer to an ECM in a state distinguished from normal physiological ECM, in which the generation, degradation, or reorganization of the ECM is abnormally regulated by disease progression or external stimuli, thereby causing tissue rigidity, increased ECM accumulation, decreased tissue elasticity, or reduced drug delivery. More specifically, the pathological extracellular matrix (ECM) described above may be induced by at least one of the following: a pathological extracellular matrix caused by ECM modification induced by factors secreted from cells or stromal cells as the disease progresses, and tissue hardening induced by repeated administration of external drugs.

[0046] In the present invention, the disease may refer to one or more diseases selected from cancer, degenerative brain disease, diabetes, chronic inflammatory disease, and metabolic disease in which a decrease in drug absorption rate occurs due to pathological ECM. Specifically, cancers that form an ECM barrier by CAF, etc. include pancreatic cancer, breast cancer, prostate cancer, lung cancer (non-small cell lung cancer and small cell lung cancer), colorectal cancer, stomach cancer, liver cancer, ovarian cancer, melanoma, glioblastoma, cholangiocarcinoma, head and neck squamous cell carcinoma, esophageal cancer, kidney cancer, bladder cancer, etc. Degenerative brain diseases are characterized by the occurrence of pathological ECM due to neuroinflammation, microglia activation, and changes in the blood-brain barrier, and include Parkinson's disease, Alzheimer's disease, Lou Gehrig's disease (amyotrophic lateral sclerosis (ALS)), Huntington's disease, multiple system atrophy, progressive supranuclear palsy, frontotemporal dementia, vascular dementia, microdegenerative disease, Creutzfeldt-Jakob disease, Parkinson's disease dementia, and Lewy body dementia. Diabetes is characterized by the occurrence of pathological ECM due to the accumulation of inflammatory substances within the ECM, and includes type 1 diabetes, type 2 diabetes, and complications of diabetes such as diabetic nephropathy, diabetic retinopathy, diabetic neuropathy, diabetic cardiomyopathy, diabetic foot ulcers, insulin resistance syndrome, non-alcoholic fatty liver disease, and diabetic dermatopathy. Chronic inflammatory diseases are conditions in which pathological ECM may develop due to a persistent imbalance in ECM production and breakdown caused by chronic inflammation; examples include rheumatoid arthritis, osteoarthritis, Crohn's disease, ulcerative colitis, psoriasis, atopic dermatitis, chronic obstructive pulmonary disease (COPD), asthma, systemic lupus erythematosus, ankylosing spondylitis, Sjögren's syndrome, and chronic sinusitis. Metabolic diseases are conditions in which pathological ECM develops due to abnormalities in energy metabolism, hormonal imbalances, adipose tissue remodeling, and changes in the ECM; examples include metabolic syndrome, non-alcoholic fatty liver disease, hyperlipidemia, atherosclerosis, steatosis, insulin resistance syndrome, and diseases related to adipose tissue fibrosis.

[0047] In addition, the pathological ECM of the present invention may be one in which the hardening of the ECM is induced by the repeated administration of a therapeutic drug. Specifically, the drug that causes the pathological ECM may be a drug that can be administered in combination with niclosamide or a pharmaceutically acceptable salt thereof, and may include anticancer agents, treatments for neurological diseases, anti-inflammatory agents, immunomodulators, treatments for metabolic diseases and diabetes, and treatments for hormones and endocrine systems. The anticancer agents may include chemotherapy agents, targeted anticancer agents, immunotherapy agents, antibody therapies, and treatments for radiation exposure. More specifically, chemotherapy drugs include paclitaxel, docetaxel, cabazitaxel, cisplatin, carboplatin, oxaliplatin, doxorubicin, epirubicin, cyclosopharmaide, ifosfamide, 5-fluorouracil, gemcitabine, methotrexate, etc.; targeted anticancer drugs include imatinib, erlotinib, gefitinib, sorafenib, sunitinib, everolimus, temusirolimus, etc.; immunotherapy drugs include nivolumab, pembrolizumab, atezolizumab, ipilimumab, etc.; antibody therapies include bevacizumab, trastuzumab, cetuximab, etc.; and radiation therapies include high-energy X-rays, gamma rays, electron beams, proton beams, heavy ion beams, and radioisotope therapy. Treatments for neurological disorders include levodopa, pramipexol, ropinirol, selegiline, rasagiline, entacapone, tolcapone, donepezil, rivastigmine, galantamine, memantine, riluzole, idrabenone, etc. Anti-inflammatory and / or immunomodulators include ibuprofen, naproxen, diclofenac, celecoxib, prednisolone, dexamethasone, methylprednisolone, infliximab, adalimumab, etanercept, tocilizumab, secukinumab, etc. Metabolic disease and diabetes treatments include metformin, glimepiride, gliburide, sitagliptin, linagliptin, empagliflozin, dapagliflozin, liraglutide, semaglutide, etc., while hormone and endocrine treatments include bicalutamide, enzalutamide, tamoxifen, letrozole, anastrozole, leuprorelin, goserelin, etc.

[0048] The composition of the present invention may further include a mineral salt to alleviate, improve, or enhance the bioavailability of the composition for the treatment of drug side effects of niclosamide or a pharmaceutically acceptable salt thereof. Any commonly known mineral salt may be used without limitation, but specifically, it may include one or more minerals selected from calcium, iron, potassium, sodium, zinc, magnesium, copper, manganese, chromium, cobalt, selenium, and molybdenum. More specifically, the salt containing the above mineral is calcium chloride, calcium sulfate, calcium nitrate, calcium oxide, calcium hydroxide, iron chloride, iron nitrate, iron sulfate, iron hydroxide, potassium chloride, potassium nitrate, potassium hydroxide, sodium chloride, sodium nitrate, sodium hydroxide, zinc chloride, zinc nitrate, zinc sulfate, zinc oxide, zinc hydroxide, magnesium chloride, magnesium nitrate, magnesium sulfate, magnesium oxide, magnesium hydroxide, copper chloride, copper nitrate, copper sulfate, copper oxide, copper hydroxide, manganese chloride, manganese nitrate, manganese sulfate, manganese oxide, manganese hydroxide, chromium chloride, chromium nitrate, chromium sulfate, chromium oxide, chromium hydroxide, cobalt chloride, cobalt nitrate, cobalt sulfate, cobalt oxide, cobalt hydroxide, selenium chloride, selenium nitrate, selenium sulfate, selenium oxide, selenium hydroxide. There may be inorganic calcium salts such as molybdenum chloride, molybdenum oxychloride, molybdenum oxide, molybdenum bromide nitrate, molybdenum nitrate, molybdenum sulfide, molybdenum hydroxide, molybdenum oxalate, calcium oxide, calcium silicate, calcium chloride, calcium carbonate, calcium sulfate, calcium nitrate, and calcium phosphate; organic calcium salts such as calcium gluconate, calcium lactate, calcium citrate, calcium acetate, and calcium formate; high molecular weight calcium salts such as calcium glycerophosphate, calcium ascorbate, calcium dextran, calcium hydroxyapatite, calcium glucoheptonate, calcium pyruvate, and calcium maleate; calcium silicates, magnesium silicates, etc., and a combination of one or more of these may be used.

[0049] Furthermore, the present invention may provide a pharmaceutical composition characterized by preparing niclosamide in an amorphous (or incompletely crystalline) state to enhance its solubility and absorption rate in the body. Specifically, the pharmaceutical composition is characterized by further including a mineral salt or a polymer additive to amorphize the niclosamide included in it.

[0050] In addition, the polymer additive in the present invention may specifically be one or more compounds selected from polyvinylpyrrolidone-based compounds, cellulose-based compounds, poloxamer-based compounds, polyethylene glycol-based compounds, alginate-based compounds, dextrin-based compounds, sugar alcohol-based compounds, and polygamma-glutamic acid. In the present invention, the polyvinylpyrrolidone-based compounds are polyvinylpyrrolidone K10 (MW 8,000–10,000), polyvinylpyrrolidone K12 (MW 11,000–12,000), polyvinylpyrrolidone K15 (MW 14,000–18,000), polyvinylpyrrolidone K17 (MW 14,000–18,000), polyvinylpyrrolidone K18 (MW 14,000–18,000), polyvinylpyrrolidone K25 (MW 20,00–25,000), polyvinylpyrrolidone K30 (MW 30,00–40,000), polyvinylpyrrolidone K60 (MW 50,00–60,000), and polyvinylpyrrolidone It may be one or more types selected from the group consisting of K90 (MW 80,000~90,000).

[0051] In the above, MW stands for molecular weight, which means weight-average molecular weight. In addition, the above poloxamer-based compounds include poloxamer 101, poloxamer 105, poloxamer 105 benzoate, poloxamer 108, poloxamer 122, poloxamer 123, poloxamer 124, poloxamer 181, poloxamer 182, poloxamer 182 dibenzoate, poloxamer 183, poloxamer 184, poloxamer 185, poloxamer 188, poloxamer 212, poloxamer 215, poloxamer 217, poloxamer 231, poloxamer 234, poloxamer 235, poloxamer 237, poloxamer 238, poloxamer 282, poloxamer 284, and poloxamer 288. It may be one or more selected from the group consisting of poloxamer 331, poloxamer 333, poloxamer 334, poloxamer 335, poloxamer 338, poloxamer 401, poloxamer 402, poloxamer 403, and poloxamer 407, and poloxamer 407 may be more preferred. The weight average molecular weight of the above-described poloxamer-based compound may be 5,000 to 500,000. In addition, the polyethylene glycol-based compound may be one or more selected from the group consisting of polyethylene glycol 200, polyethylene glycol 300, polyethylene glycol 400, polyethylene glycol 500, polyethylene glycol 1000, polyethylene glycol 1400, polyethylene glycol 1500, polyethylene glycol 4000, polyethylene glycol 8000, polyethylene glycol 10000, and methoxypolyethylene glycol 550. The weight average molecular weight of the polyethylene glycol-based compound described above may be 5,000 to 500,000. The alginate-based compound may be an alginate, alginic acid, and alginate ester, etc., and the alginate may be sodium alginate, ammonium alginate, calcium alginate lactate, potassium alginate, etc.The above dextrin-based compounds may be sodium dextrin sulfate, matodextrin, icodextrin, amylodextrin, α-cyclodextrin, β-cyclodextrin, γ-cyclodextrin, hydroxypropyl-α-cyclodextrin, hydroxypropyl-βcyclodextrin, hydroxypropyl-γ-cyclodextrin, dextran, dextran40, dextran70, etc. The above sugar alcohol compound may be a compound of HOCH2(CHOH)nCH2OH, where n is an integer from 1 to 30, and more specifically, it may be ethylene glycol, glycerol, erythritol, threitol, arabitol, xylitol, ribitol, mannitol, sorbitol, galactitol, fucitol, iditol, inositol, bolemitol, isomalt, maltitol, lactitol, maltotritol, maltotetraitol, and polyglycitol, etc.

[0052] The composition of the present invention may comprise 0.1 to 50 weight% of niclosamide or a pharmaceutically acceptable salt thereof and 0.001 to 50 weight% of a mineral salt compound, when the total weight of the composition is 100 weight%.

[0053] In addition, the present invention relates to niclosamide or a pharmaceutically acceptable salt thereof.

[0054] The present invention provides a method for preparing a niclosamide pharmaceutical composition comprising the steps of: (a) mixing with a polymer additive and a mineral salt; (b) stirring under an organic solvent (such as ethanol) or a water-ethanol mixed solvent; and (c) preparing the composition in an amorphous (or incompletely crystalline) state through low-temperature drying or vacuum drying. It is evident that the order of steps (a) to (c) described above may be changed as necessary. When a pharmaceutical composition containing niclosamide or a pharmaceutically acceptable salt thereof is prepared by the method described above, it can be prepared in an amorphous, incompletely crystalline, or amorphous state, thereby significantly improving solubility and bioavailability compared to crystalline niclosamide.

[0055] The composition of the present invention may additionally include ingredients that are commonly used in pharmaceutical compositions to improve odor, taste, visual appearance, etc., although they do not increase the medicinal effect. Additionally, the pharmaceutical composition of the present invention may additionally include pharmaceutically acceptable additives. Pharmaceutically acceptable additives include, but are not limited to, starch, gelatinized starch, microcrystalline cellulose, lactose, povidone, colloidal silicon dioxide, calcium hydrogen phosphate, lactose, mannitol, malt syrup, gum arabic, pregelatinized starch, corn starch, powdered cellulose, hydroxypropyl cellulose, Opadry, sodium starch glycolate, carnauba wax, synthetic aluminum silicate, stearic acid, magnesium stearate, aluminum stearate, calcium stearate, sucrose, dextrose, sorbitol, and talc. The pharmaceutical composition of the present invention may further include a pharmaceutically acceptable carrier and may be formulated for human or veterinary use for oral or parenteral administration. When formulating the pharmaceutical composition of the present invention, in addition to the components listed above, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants may be used. Solid dosage forms for oral administration include tablets, pills, powders, granules, and capsules, and such solid dosage forms may be prepared by mixing at least one excipient, for example, starch, calcium carbonate, sucrose or lactose, and gelatin mannitol, with the pharmaceutical composition containing the compound of the present invention. In addition, lubricants such as magnesium stearate and talc may be used in addition to simple excipients. Liquid formulations for oral administration include suspensions, liquid formulations, emulsions, and syrups. In addition to commonly used simple diluents such as water and liquid paraffin, various excipients, such as humectants, sweeteners, flavorings, and preservatives, may be used.

[0056] The composition of the present invention may be administered orally or parenterally depending on the intended method, and when administered parenterally, it is preferable to select a method of external application to the skin or intraperitoneal injection, rectal injection, subcutaneous injection, intravenous injection, intramuscular injection, or intrathoracic injection.

[0057] In addition, the present invention provides a method of administration for preventing the formation of pathological extracellular matrix (ECM) or alleviating or improving a formed pathological extracellular matrix, comprising the step of orally administering a composition containing niclosamide or a pharmaceutically acceptable salt thereof as an active ingredient. Specifically, the step of oral administration may involve administering niclosamide or a pharmaceutically acceptable salt thereof multiple times. More specifically, the administration of niclosamide or a pharmaceutically acceptable salt thereof may be administered in the range of 10 to 200 mg / kg / day, and multiple administration may involve administering niclosamide or a pharmaceutically acceptable salt thereof two or more times a day. When administered within the aforementioned dosage range and frequency, niclosamide or its pharmaceutically acceptable salts can effectively prevent, alleviate, or improve the formation, accumulation, and hardening of the pathological extracellular matrix (ECM) induced by disease progression or repeated administration of external drugs by maintaining stable in vivo exposure even during long-term and repeated treatment courses. In particular, by continuously exhibiting regulatory effects on the pathological ECM while lowering the risk of side effects compared to high-dose single administration, it provides the effect of improving treatment continuity and compliance in various diseases requiring long-term treatment.

[0058] In the present invention, the pathological extracellular matrix (ECM) may be induced by at least one of the pathological extracellular matrix caused by ECM modification induced by factors secreted from cells or stromal cells as the disease progresses, and tissue hardening induced by repeated administration of external drugs.

[0059] More specifically, the method of administration for preventing the formation of pathological extracellular matrix (ECM) according to the present invention, or for alleviating or improving the formed pathological extracellular matrix, may involve reducing the expression level of one or more of the MMP-9 and VEGF factors. More specifically, the MMP-9 and VEGF factors may mean that the expression level of MMP-9 and / or VEGF in the group administered the composition of the present invention is reduced by at least 10% compared to the vehicle administration group after pathology induction, and more preferably, MMP-9 may be reduced by at least 30% and VEGF by at least 20%.

[0060] In addition, the present invention provides a combination administration method for preventing the formation of pathological extracellular matrix (ECM) or alleviating or improving a formed pathological extracellular matrix, comprising: a pretreatment step of orally administering a composition containing niclosamide or a pharmaceutically acceptable salt thereof as an active ingredient for 3 to 7 days prior to drug administration; and a step of administering the drug in combination with the composition after the pretreatment step. When administered by the administration method proposed by the present invention, it may be applied to anticancer treatment for prostate cancer, triple-negative breast cancer, metastatic sarcoma, hematological cancer, pancreatic cancer, or other cancers in which ECM remodeling plays a major role, or to the treatment of side effects caused by the administration of anticancer drugs. Furthermore, the pharmaceutical composition and / or treatment method of the present invention not only prevent cancer metastasis but also control and inhibit cancer-induced ECM formation, thereby enabling the prevention of the occurrence of cancer itself through appropriate pre-administration.

[0061] Specifically, in the pretreatment step, the composition may be administered in a range of 10 to 200 mg / kg / day until the expression level of one or more of the MMP-9 and VEGF factors is reduced. By inducing a state in which the expression levels of MMP-9 and / or VEGF are reduced in the pretreatment step, the delivery of the drug to the tissue subsequently administered in combination can be improved, and the hardening and re-strengthening of the extracellular matrix induced by repeated administration can be suppressed, thereby maintaining or enhancing the therapeutic effect.

[0062]

[0063] In addition, a composition comprising niclosamide or a pharmaceutically acceptable salt thereof as an active ingredient may further comprise a mineral salt.

[0064] In addition, the present invention comprises the step of administering a drug in combination with a composition comprising niclosamide or a pharmaceutically acceptable salt thereof as an active ingredient,

[0065] The present invention provides a method of concomitant administration for preventing the formation of pathological extracellular matrix (ECM) or for alleviating or improving a formed pathological extracellular matrix. The aforementioned oral administration step may involve administering niclosamide or a pharmaceutically acceptable salt thereof multiple times, and the specific dosage may be administered in the range of 10 to 200 mg / kg / day, and the aforementioned multiple administration may involve administering niclosamide or a pharmaceutically acceptable salt thereof at least twice a day.

[0066] The drugs used for combination administration in the present invention may include anticancer agents, treatments for nervous system diseases, anti-inflammatory agents, immunomodulators, treatments for metabolic diseases and diabetes, hormone and endocrine system treatments, etc., and anticancer agents may include chemotherapy agents, targeted anticancer agents, immunotherapy agents, antibody treatments, and treatments for radiation exposure. More specifically, chemotherapy drugs include paclitaxel, docetaxel, cabazitaxel, cisplatin, carboplatin, oxaliplatin, doxorubicin, epirubicin, cyclosopharmaide, ifosfamide, 5-fluorouracil, gemcitabine, methotrexate, etc.; targeted anticancer drugs include imatinib, erlotinib, gefitinib, sorafenib, sunitinib, everolimus, temusirolimus, etc.; immunotherapy drugs include nivolumab, pembrolizumab, atezolizumab, ipilimumab, etc.; antibody therapies include bevacizumab, trastuzumab, cetuximab, etc.; and radiation therapies include high-energy X-rays, gamma rays, electron beams, proton beams, heavy ion beams, and radioisotope therapy. Treatments for neurological disorders include levodopa, pramipexol, ropinirol, selegiline, rasagiline, entacapone, tolcapone, donepezil, rivastigmine, galantamine, memantine, riluzole, idrabenone, etc. Anti-inflammatory and / or immunomodulators include ibuprofen, naproxen, diclofenac, celecoxib, prednisolone, dexamethasone, methylprednisolone, infliximab, adalimumab, etanercept, tocilizumab, secukinumab, etc. Metabolic disease and diabetes treatments may include metformin, glimepiride, gliburide, sitagliptin, linagliptin, empagliflozin, dapagliflozin, liraglutide, semaglutide, etc., and hormone and endocrine treatments may include bicalutamide, enzalutamide, tamoxifen, letrozole, anastrozole, leuprorelin, goserelin, etc.

[0067] Each step of the combination administration method and pretreatment method described in the present invention is not necessarily limited to the order described, and the order may be appropriately changed depending on the condition of the subject to treatment, the type of disease, the characteristics of the combination drugs, or clinical necessity. Furthermore, the administration order, dosage, duration of administration, and combination presented in the present invention may be interchanged or modified within the scope of the technical spirit of the present invention, and such modifications should be understood as obvious to those skilled in the art.

[0068] Example 1: Niclosamide (Manufacturing, testing, release and stability testing site, Company Name: Derivados Quimicos SAU; DMS Holder's Company Name: Olon S. p. A,)

[0069] Example 2 (CP-EFR, Penetrium): 2 g of hydroxycellulose (HPMC6) and 80% ethanol solvent were added to a reactor and dissolved by stirring. Subsequently, 40 g of niclosamide was added, stirred, and dried to prepare a primary mixture. The primary mixture and 22.24 g of magnesium oxide were placed in a mixer and wet-mixed using anhydrous ethanol, followed by drying to prepare a secondary mixture. The dried secondary mixture and excipients were dry-mixed using a drum mixer and filled into gelatin capsules (No. 4). The specific composition of the mixture is shown in Table 1.

[0070] CP-EFR Raw Material Quantity Comparison Table X-05 Raw Material Name Prescription Composition Ratio (mg) (%) Niclosamide 40.00 40.00 Magnesium oxide 22.24 22.24 HPMC 62.00 2.00 SMCC 9.02 9.02 LHPC 10.24 10.24 Anhydrous lactose 15.50 15.50 Magnesium stearate 1.00 1.00 Total 100.00 100.00

[0071] Example 3: 2g of hydroxycellulose (HPMC6) and 80% ethanol solvent were added to a reactor and stirred to dissolve, then 40g of niclosamide was added and stirred and dried to prepare a primary mixture.

[0072] Experimental Example 1: Administration of Niclosamide to a Patient Dog (Poodle) with Anticancer Drug Side Effects

[0073] Two individuals (A and B) that developed cancer were administered the anticancer drug polytaxel three times at 7-day intervals, and their progress was observed for 8 to 12 weeks. Both individuals that developed cancer were Poodles. In all individuals, hardening of the tumor site was observed as a side effect after polytaxel administration. To compare the difference between the experimental group (A) and the control group (B), individual A (experimental group) was additionally administered Example 2 (CP-EFR), a formulation containing niclosamide along with polytaxel, and individual B (control group) was administered polytaxel alone, and their progress was observed for the same period.

[0074] Specifically, Subject A (experimental group) was administered Example 2 (CP-EFR) at a dose of 40 mg / head twice a day for 5 days starting from December 4, 2024, after the onset of side effects on December 3, 2024, and the anticancer drug Polytaxel was re-administered starting from December 9, 2024, following the administration of Example 2. During re-administration, Polytaxel was administered 3 times at 7-day intervals, and starting from December 12, 2024, Example 2 (CP-EFR) was administered at a dose of 40 mg / head twice a day.

[0075] The dosage and duration of administration of polytaxel administered to the two subjects are as follows.

[0076] [Polytaxel dose(mg / kg)]

[0077] Patient A 24.9.5 ~ 24.9.19: 4.5 mg / kg (Docetaxel dose 0.51 mg / kg)

[0078] Dog B 24.10.4 ~ 24.10.17: 6.5 mg / kg (Docetaxel dose 0.73 mg / kg)

[0079] A patient 24.12.09 ~ 24.12.23: 5.5 mg / kg (Docetaxel dose 0.62 mg / kg)

[0080] Dog B 24.12.09 ~ 24.12.23: 5.5 mg / kg (Docetaxel dose 0.62 mg / kg)

[0081] Specific details regarding the experimental method are shown in Fig. 1, and the experimental results for individual A and individual B are shown in Figs. 2 and 3, respectively. Upon reviewing the results, it was confirmed that individual A, who was administered the composition of Example 2, showed an effect of the hardened carcinoma becoming soft on December 9, 2024, which is 5 days after December 4, 2024, when the composition of Example 2 was administered. The hardness of the tumor upon palpation is listed in Table 2, and the softening effect of the carcinoma is listed in Table 3.

[0082] 1 (Very Soft) Similar to normal soft tissue; it sinks easily when pressed with a finger and slowly returns to its original state. 2-3 (Soft) Soft but with slight resistance. It is fluid and moves easily upon palpation. 4-5 (Medium Hardness) Elastic upon palpation with moderate resistance. 6-7 (Hard) The tumor has clear boundaries and is difficult to move. It has strong rebound force upon pressing and low elasticity. 8-9 (Very Hard) Adhered to surrounding tissues and hardly moves. It hardly deforms even when pressure is applied during palpation. 10 (Rock-like Hardness) Like hard bone tissue, it hardly compresses. It does not deform even when pressed hard with a finger.

[0083]

[0084] A Patient (Samsuni) B Patient (Choco) 2024.12.4 (Before CP-EFR administration) 109 2024.12.9 (After CP-EFR administration) 29

[0085] As can be seen from the results in Table 3, in Subject A, who was administered the pharmaceutical composition of Example 2, the tumor hardened due to side effects softened, and the tumor was barely palpable. From this, it can be inferred that the problem of ECM hardening, the external environment of the tumor, was improved. Furthermore, as a result, it was confirmed that the tumor size decreased by more than 70% by December 30, 2024, the end date of administration, confirming that the side effects of the anticancer drug were alleviated and the therapeutic effect of the anticancer drug was enhanced. On the other hand, Subject B, who was administered the anticancer drug alone, showed a tumor size reduction rate of less than 50%, and in certain areas, the tumor actually grew larger, confirming that not only was the therapeutic efficiency of the anticancer drug lowered, but the side effects of the anticancer drug were also intensified.

[0086] As a result of this experiment, it was confirmed that administering Example 2 (CP-EFR; Penetrium) alleviated tumor sclerosis caused by ECM sclerosis, a side effect of the anticancer drug (polytaxel), and increased the therapeutic effect of the anticancer drug. On the other hand, when the anticancer drug was administered alone, there was a tendency for side effects to worsen and the therapeutic effect to decrease. From these results, it was confirmed that the composition containing niclosamide alleviated drug side effects and maximized the therapeutic efficiency of the anticancer drug.

[0087] Experimental Example 2. Changes in MMP9 and VEGF expression observed after lung excision in a syngeneic mouse model (C57BL / 6 mouse) transplanted with an LL2 tumor, administered Bevacizumab (5 mg / kg, QW) alone or in combination with CP-EFR 50, 100, and 150 mg / kg.

[0088] In this experiment, a syngeneic model was constructed by subcutaneously implanting the LL / 2 cell line, a lung cancer cell line, into C57BL / 6 mice, and then tumor size was verified and the cell profile was observed.

[0089] The composition of Example 2, which contains niclosamide, was administered to suppress anticancer drug side effects through various signaling pathways in cells and to induce a synergistic effect of the anticancer drug; Bevacizumab (hereinafter referred to as (B)), which inhibits tumor angiogenesis, was selected and administered as the anticancer drug. (B) was selected because it targets the VEGF mechanism and was judged to exhibit a synergistic effect with niclosamide. The specific experimental details are as follows.

[0090] Experimental conditions

[0091] (1) Mouse: C57BL / 6NHsd mouse female, 5 weeks old

[0092] (2) Cell line: LL / 2 NSCLC cell line

[0093] Details of experimental conditions

[0094] Immunotherapy-resistant, VEGF-overexpressing cell lines

[0095] test material

[0096] Test drug: Example 2 (CP-EFR) (50, 100, 150, 200 mg / kg)

[0097] Concomitant drug: Bevacizumab(B) (5 mg / kg)

[0098] The administration method for each group is as follows.

[0099] G1: No treatment group

[0100] G4: Bevacizumab monotherapy group

[0101] G13: CP-EFR 50mg / kg(TID)+ Bevacizumab

[0102] G14: CP-EFR 100mg / kg(TID)+ Bevacizumab

[0103] G15: CP-EFR 150mg / kg(TID)+ Bevacizumab

[0104] Group Administered Substance Dosage Frequency of Administration ( / week) Number of Animals (B) CP-EFR (Example 2) G4 Bevacizumab (B) 50 26 G13 Bevacizumab+CP-EFR 55 0 26 G14 Bevacizumab+ CP-EFR 51 0 26 G15 Bevacizumab+ CP-EFR 51 5 0 26

[0105] Method of administration

[0106] Excipients: 0.5% MC

[0107] Administered volume: 10 mL / kg

[0108] Dosage interval: Orally 3 times a day (8-hour intervals)

[0109] Concurrent drug administration: Concurrent administration within 1 to 2 hours after administration of the Example 2 composition (CP-EFR).

[0110] Date acquired: 2024 / 10 / 02 (Female acquired at 5 weeks old)

[0111] Date of administration start: 2024 / 10 / 18 (Day 11 of engraftment)

[0112] End of administration date: 2024 / 11 / 04 (17th day of administration)

[0113]

[0114] The results of Experimental Example 2, obtained through the test process described above, are shown in FIGS. 4, 5, and 6. As can be seen from the experimental results, in groups G13 to G15 treated with Example 2 (CP-EFR), it was confirmed that metastasis did not occur because circulating tumor cells (CTCs) could not settle in the organ (lung).

[0115]

[0116] In the control group, lung metastases were observed in 5 out of 6 animals, and in the Bevacizumab monotherapy group, lung metastases were observed in 4 out of 6 animals. On the other hand, in the Bevacizumab and CP-EFR 50 mg / kg combination therapy group, lung metastases were observed in only 2 out of 6 animals, and in the Bevacizumab and CP-EFR 100 mg / kg or 150 mg / kg combination therapy groups, no lung metastases were observed.

[0117] In addition, specifically, in a C57BL / 6 mouse syngeneic model transplanted with LL / 2 lung cancer cell lines, compared to the group administered Bevacizumab alone (G4), the MMP-9 signal intensity in the lung tissue of the groups administered a composition containing niclosamide (CP-EFR) in combination (G13, G14, G15) was reduced to approximately 12.5 AU (G13), 11.8 AU (G14), and 12.5 AU (G15) compared to an average of approximately 15.9 AU (G4), which corresponds to a reduction of approximately 20% to 50% compared to the control group.

[0118] In addition, under the same experimental conditions, the VEGF signal intensity decreased to approximately 6.2 AU in the group administered CP-EFR 100 mg / kg (TID) in combination (G14) compared to an average of approximately 12.8 AU in the group administered Bevacizumab alone (G4), showing a decrease of more than 50%, and also decreased to approximately 9.1 AU in the group administered CP-EFR 150 mg / kg (TID) in combination (G15), showing a decrease of more than 20%.

[0119]

[0120]

[0121] Experimental Example 3

[0122] SH-SY5Y cells (Passage 5) were cultured in 5 x 10 wells per well in a 96-well plate (SPL Culture Plate #30096). 4Cells were seeded at cellular densities and cultured in EMEM medium supplemented with 10% FBS. Niclosamide was administered after an additional 24 hours of incubation for attachment and stabilization. A total of 11 concentrations of niclosamide (Derivados Quimicos, Batch#140002M02K) were prepared by serially diluting 1 / 3 of a concentration, starting with a peak of 10 μM; among these, six concentrations in the low-concentration range, including a vehicle control (0 μM), were selected for treatment. After 24 hours following niclosamide treatment, MPP was applied to induce a cell damage model. + 1 (Sigma-Aldrich, Cat#D048) was added to make the concentration 1 mM. MPP + An MTT assay was performed to evaluate cell viability after 24 hours of treatment. After adding MTT solution to each well, the reaction was allowed to proceed for 2 hours, and the cell viability was analyzed by measuring the absorbance of the generated formazan at 570 nm, as shown in Figure 7.

[0123] MTT analysis results, MPP + In the group treated alone, cell viability decreased to approximately 45.1%, indicating significant cytotoxicity in SH-SY5Y cells; therefore, it can be concluded that an inflammatory state was induced. Under these conditions, the group pretreated with Niclosamide showed a tendency for cell viability to recover to a range of approximately 60.7–68.5%. Notably, at certain concentrations, viability increased to 64–68%, suggesting that Niclosamide exhibits a protective effect in mitigating MPP-induced cell damage. Overall, the Niclosamide-treated group [applied to] MPP + A consistent increase in survival rate was observed compared to the single group, and a gradual recovery trend was observed with increasing concentration, suggesting a partial concentration-dependent cytoprotective effect.

[0124]

[0125] Experimental Example 4-1

[0126] EO771 cells, a mouse-derived breast cancer cell line, were injected into 7-week-old female C57BL / 6 mice at a rate of 5×10 5 The efficacy of CP-EFR and Paclitaxel combination therapy was evaluated by subcutaneous injection.

[0127] experimental group

[0128] Group 1: Vehicle

[0129] Group 2: Paclitaxel (2 mg / kg), IP, EOD

[0130] Group 3: CP-EFR (100 mg / kg)

[0131] Group 4: CP-EFR (50 mg / kg), PO, BID + Paclitaxel (2 mg / kg), IP, EOD

[0132] Group 5: CP-EFR (100 mg / kg), PO, BID + Paclitaxel (2 mg / kg), IP, EOD

[0133] Group 6: CP-EFR (200 mg / kg), PO, QD + Paclitaxel (2 mg / kg), IP, EOD

[0134] * PO: Oral administration

[0135] * IP: Intraperitoneal administration

[0136] * QD: Administer once daily

[0137] * BID: Administer twice daily (12-hour intervals)

[0138] * EOD: Administer once every 2 days

[0139]

[0140] However, when administered in combination, the first oral administration was followed by Paclitaxel intraperitoneal administration 6 hours later, and the second oral administration 12 hours later.

[0141] The long and short axes of the tumor were measured using calipers every two days starting from the start of administration, and the tumor size was calculated using the following formula.

[0142] Tumor volume calculation formula

[0143] a = longest length of the tumor

[0144] b = shortest length of the tumor

[0145] Total tumor size = (a × b²) / 2

[0146] The results of the above experiment are shown in Figure 8.

[0147] As can be seen in Figure 8, no statistically significant anticancer efficacy was observed in the Paclitaxel monotherapy group (Group 2) compared to the vehicle control group, which suggests that the ECM was altered by repeated administration of Paclitaxel, preventing the drug from effectively penetrating.

[0148] On the other hand, in the group administered alone of the pharmaceutical composition of Example 2 (CP-EFR) (Group 3) and the group administered in combination with Paclitaxel and the pharmaceutical composition of Example 2 (CP-EFR) (Groups 4, 5, and 6), Paclitaxel was effectively delivered to cancer cells, and statistically significant anticancer efficacy was confirmed. In particular, the group administered with CP-EFR 100 mg / kg twice daily and the group administered with CP-EFR 200 mg / kg once daily showed a statistically significant difference compared to the group administered with Paclitaxel alone.

[0149] Experimental Example 4-2. Mouse EO771 Cell Line Lung Metastatic Cancer Model Experiment

[0150] 5 x 10 cells of the mouse-derived breast cancer cell line EO771 were injected into 7-week-old female C57BL / 6 mice. 5

[0151] The anti-metastatic efficacy of the combination therapy of the composition of Example 2 (CP-EFR) and Paclitaxel was evaluated by administering it via microvenous injection.

[0152] - Group 1: Vehicle

[0153] - Group 2: Paclitaxel 2 mg / kg, IP, EOD

[0154] - Group 3: Composition of Example 2 (CP-EFR) 50 mg / kg, PO, BID + Paclitaxel (2 mg / kg), IP, EOD

[0155] Gross observation of the lung tissue in Figure 9 confirmed that in the group treated with Paclitaxel alone (Group 2), metastatic nodules on the lung surface increased significantly compared to the excipient control group (Group 1), and the pattern of metastasis was accelerated. This suggests that repeated administration of paclitaxel induced the abnormal reorganization and accumulation of pathological extracellular matrix (ECM) within the lung tissue, thereby forming a 'pre-metastatic niche,' an environment favorable for circulating cancer cells to settle. In other words, it confirmed the side effect that the pathological ECM induced by the administration of the anticancer drug itself acts as a barrier and soil that facilitates the metastasis of cancer cells.

[0156] On the other hand, in the group administered with Paclitaxel and the pharmaceutical composition of the present invention (CP-EFR) (Group 3), the formation of metastatic nodules on the surface of the lungs was strongly inhibited to the extent that they were hardly visible to the naked eye. This demonstrates that CP-EFR fundamentally prevents cancer cells from adhering to lung tissue by preemptively preventing the formation of pathological ECM induced by paclitaxel or by alleviating and improving the rigidity of the abnormally formed pathological extracellular matrix.

[0157] Consequently, the composition of the present invention effectively controls pathological ECM deformation of tissues caused by external drug administration to maintain tissue homeostasis, thereby overcoming the metastasis-promoting side effects that occur when an anticancer drug is administered alone.

[0158] Experimental Example 5 Mouse (Nude) Test

[0159] Mice in a total of four groups were administered docetaxel (PO) or docetaxel (DTX; PO) and the niclosamide composition of Example 3 (NIC; PO) in the following amounts and schedules, and the results regarding changes in body weight are shown in Figures 10 and 11.

[0160] Oral administration of G1 DTX 20 mg / kg to mice / Administered twice daily at 12-hour intervals for 8 days

[0161] Oral administration of G2 DTX 40 mg / kg to mice / Administered twice daily at 12-hour intervals for 8 days

[0162] Oral administration of G3 DTX 20 mg / kg to mice / Administered twice daily at 12-hour intervals for 8 days

[0163] Oral administration of NIC 100 mg / kg to mice / Administered twice daily at 12-hour intervals for 8 days

[0164] Oral administration of G4 DTX 20 mg / kg to mice / Administered twice daily at 12-hour intervals for 8 days

[0165] Oral administration of NIC 100 mg / kg to mice / Administered twice daily at 12-hour intervals for 8 days

[0166] As can be seen in Figures 10 and 11, significant weight loss was observed in the groups administered anticancer drugs alone (G1, G2), while it was confirmed that the side effects caused by weight loss were recovered in the groups administered niclosamide in combination (G3, G4). When administering anticancer drugs, most patients may experience cachexia as a side effect; this side effect reduces physical strength and causes metabolic abnormalities, thereby posing a problem that lowers the anticancer effect. The weight recovery effect observed in the G3 and G4 groups suggests that niclosamide is effective in overcoming cachexia, a side effect of anticancer drugs, and proposes niclosamide as a strategy to reduce the side effects of anticancer drugs.

[0167] This result supports the fact that administering is effective.

[0168] Experimental Example 6 Mouse (Nude) Test

[0169] In a total of four groups, mice were injected with docetaxel (IV) or docetaxel (DTX; IV) and orally administered the niclosamide composition (NIC; PO) of Example 2 for 28 days, and the results regarding changes in body weight after administration are shown in Figures 12 and 13.

[0170] Administer G1 DTX 2 mg / kg to mice via IV / Administer at a concentration of 10 ml / kg once daily on days 0, 4, and 7

[0171] Administer G2 DTX 2 mg / kg to mice by injection (IV) / Administer at a concentration of 10 ml / kg once daily on days 0, 4, and 7, administer NIC 200 mg / kg to mice orally / Administer twice daily at 12-hour intervals for 28 days

[0172] The same results as in Experimental Example 5 were confirmed in Figures 12 and 13. It was confirmed that significant weight loss occurred in the group administered the anticancer drug alone (G1), and that the side effects caused by weight loss were recovered in the group administered niclosamide in combination (G2). In other words, the weight recovery effect observed in the G2 group suggests that it is effective in overcoming cachexia, a side effect of the anticancer drug, and supports the result that administering niclosamide is an effective strategy to reduce the side effects of the anticancer drug.

Claims

1. A composition for preventing the formation of pathological extracellular matrix (ECM), or for alleviating or improving a formed pathological extracellular matrix, comprising niclosamide or a pharmaceutically acceptable salt thereof as an active ingredient.

2. A composition for preventing the formation of a pathological extracellular matrix (ECM) or alleviating or improving a formed pathological extracellular matrix, wherein the pathological extracellular matrix (ECM) is induced by at least one of a pathological extracellular matrix caused by ECM modification induced by factors secreted from cells or stromal cells as the disease progresses, and tissue hardening induced by repeated administration of an external drug.

3. A composition for preventing the formation of pathological extracellular matrix (ECM) or alleviating or improving a formed pathological extracellular matrix, wherein the disease of claim 2 is one or more diseases selected from cancer, degenerative brain disease, diabetes, chronic inflammatory disease and metabolic disease in which a decrease in drug absorption rate occurs due to pathological ECM.

4. A composition for preventing the formation of pathological extracellular matrix (ECM), or alleviating or improving a formed pathological extracellular matrix, wherein the drug of claim 1 is a drug capable of causing hardening of the ECM due to repeated administration.

5. In Claim 4, A composition for preventing the formation of pathological extracellular matrix (ECM), or alleviating or improving a formed pathological extracellular matrix, wherein the drug capable of causing sclerosis of the ECM due to repeated administration is one or more selected from anticancer agents, treatments for neurological diseases, anti-inflammatory agents, and immunomodulators.

6. In claim 1, the composition further comprises a mineral salt, and The above mineral salts are calcium chloride, calcium sulfate, calcium nitrate, calcium oxide, calcium hydroxide, iron chloride, iron nitrate, iron sulfate, iron hydroxide, potassium chloride, potassium nitrate, potassium hydroxide, sodium chloride, sodium nitrate, sodium hydroxide, zinc chloride, zinc nitrate, zinc sulfate, zinc oxide, zinc hydroxide, magnesium chloride, magnesium nitrate, magnesium sulfate, magnesium oxide, magnesium hydroxide, copper chloride, copper nitrate, copper sulfate, copper oxide, copper hydroxide, manganese chloride, manganese nitrate, manganese sulfate, manganese oxide, manganese hydroxide, chromium chloride, chromium nitrate, chromium sulfate, chromium oxide, chromium hydroxide, cobalt chloride, cobalt nitrate, cobalt sulfate, cobalt oxide, cobalt hydroxide, selenium chloride, selenium nitrate, selenium sulfate, selenium oxide, selenium hydroxide, molybdenum chloride, A composition for preventing the formation of a pathological extracellular matrix (ECM), or alleviating or improving a formed pathological extracellular matrix, comprising one or more selected from molybdenum oxychloride, molybdenum oxide, molybdenum bromide nitrate, molybdenum nitrate, molybdenum sulfide, molybdenum hydroxide, molybdenum oxalate, calcium oxide, calcium silicate, calcium chloride, calcium carbonate, calcium sulfate, calcium nitrate, calcium phosphate, calcium gluconate, calcium lactate, calcium citrate, calcium acetate, organic calcium salts such as calcium foamate, calcium glycerophosphate, calcium ascorbate, calcium dextran, calcium hydroxyapatite, calcium glucoheptonate, calcium pyruvate, calcium maleate, calcium silicate, and magnesium silicate.

7. A composition for preventing the formation of pathological extracellular matrix (ECM), or alleviating or improving a formed pathological extracellular matrix, wherein the composition of claim 1 further comprises a polymer additive.

8. A method of administration for preventing the formation of pathological extracellular matrix (ECM) or alleviating or improving a formed pathological extracellular matrix, comprising the step of orally administering a composition comprising niclosamide or a pharmaceutically acceptable salt thereof as an active ingredient.

9. In Claim 8, The above oral administration step is a method of administration for preventing the formation of pathological extracellular matrix (ECM) or alleviating or improving a formed pathological extracellular matrix, wherein niclosamide or a pharmaceutically acceptable salt thereof is administered multiple times.

10. In Claim 9, A method of administration for preventing the formation of pathological extracellular matrix (ECM) or alleviating or improving a formed pathological extracellular matrix, wherein the dosage of the above-mentioned niclosamide or its pharmaceutically acceptable salt is administered in the range of 10 to 200 mg / kg / day.

11. In Claim 9, The above multiple administration is a method of administration for preventing the formation of pathological extracellular matrix (ECM) or alleviating or improving a formed pathological extracellular matrix, wherein niclosamide or a pharmaceutically acceptable salt thereof is administered at least twice a day.

12. In claim 8, A method of administration for preventing the formation of a pathological extracellular matrix (ECM) or alleviating or improving a formed pathological extracellular matrix, wherein the pathological extracellular matrix (ECM) is induced by at least one of a pathological extracellular matrix caused by ECM modification induced by factors secreted from cells or stromal cells as the disease progresses, and tissue hardening induced by repeated administration of an external drug.

13. In claim 8, A method of administration for preventing the formation of the above-mentioned pathological extracellular matrix (ECM) or for alleviating or improving the pathological extracellular matrix is ​​a method of administration for preventing the formation of the pathological extracellular matrix (ECM) or for alleviating or improving the pathological extracellular matrix, wherein the method reduces the expression level of one or more of the MMP-9 and VEGF factors.

14. In Claim 8, The composition comprising the above-mentioned niclosamide or a pharmaceutically acceptable salt thereof as an active ingredient further comprises a mineral salt, and The above mineral salts are calcium chloride, calcium sulfate, calcium nitrate, calcium oxide, calcium hydroxide, iron chloride, iron nitrate, iron sulfate, iron hydroxide, potassium chloride, potassium nitrate, potassium hydroxide, sodium chloride, sodium nitrate, sodium hydroxide, zinc chloride, zinc nitrate, zinc sulfate, zinc oxide, zinc hydroxide, magnesium chloride, magnesium nitrate, magnesium sulfate, magnesium oxide, magnesium hydroxide, copper chloride, copper nitrate, copper sulfate, copper oxide, copper hydroxide, manganese chloride, manganese nitrate, manganese sulfate, manganese oxide, manganese hydroxide, chromium chloride, chromium nitrate, chromium sulfate, chromium oxide, chromium hydroxide, cobalt chloride, cobalt nitrate, cobalt sulfate, cobalt oxide, cobalt hydroxide, selenium chloride, selenium nitrate, selenium sulfate, selenium oxide, selenium hydroxide, molybdenum chloride, A method of administration for preventing the formation of pathological extracellular matrix (ECM) or alleviating or improving a formed pathological extracellular matrix, wherein the agent is one or more selected from organic calcium salts such as molybdenum oxychloride, molybdenum oxide, molybdenum bromide nitrate, molybdenum nitrate, molybdenum sulfide, molybdenum hydroxide, molybdenum oxalate, calcium oxide, calcium silicate, calcium chloride, calcium carbonate, calcium sulfate, calcium nitrate, calcium phosphate, calcium gluconate, calcium lactate, calcium citrate, calcium acetate, and calcium foamate, calcium glycerophosphate, calcium ascorbate, calcium dextran, calcium hydroxyapatite, calcium glucoheptonate, calcium pyruvate, calcium maleate, calcium silicate, and magnesium silicate.

15. A pretreatment step of orally administering a composition comprising niclosamide or a pharmaceutically acceptable salt thereof as an active ingredient for 3 to 7 days prior to drug administration; and A method for co-administration to prevent the formation of pathological extracellular matrix (ECM) or to alleviate or improve a formed pathological extracellular matrix, comprising the step of administering the drug in combination with the composition after the above pretreatment step.

16. A method of combination administration for preventing the formation of pathological extracellular matrix (ECM) or alleviating or improving a formed pathological extracellular matrix, wherein, in the pretreatment step, the composition is administered in a range of 10 to 200 mg / kg / day until the expression level of one or more of MMP-9 and VEGF factors is reduced.

17. The composition of claim 15, comprising niclosamide or a pharmaceutically acceptable salt thereof as an active ingredient, further comprises a mineral salt, and The above mineral salts are calcium chloride, calcium sulfate, calcium nitrate, calcium oxide, calcium hydroxide, iron chloride, iron nitrate, iron sulfate, iron hydroxide, potassium chloride, potassium nitrate, potassium hydroxide, sodium chloride, sodium nitrate, sodium hydroxide, zinc chloride, zinc nitrate, zinc sulfate, zinc oxide, zinc hydroxide, magnesium chloride, magnesium nitrate, magnesium sulfate, magnesium oxide, magnesium hydroxide, copper chloride, copper nitrate, copper sulfate, copper oxide, copper hydroxide, manganese chloride, manganese nitrate, manganese sulfate, manganese oxide, manganese hydroxide, chromium chloride, chromium nitrate, chromium sulfate, chromium oxide, chromium hydroxide, cobalt chloride, cobalt nitrate, cobalt sulfate, cobalt oxide, cobalt hydroxide, selenium chloride, selenium nitrate, selenium sulfate, selenium oxide, selenium hydroxide, molybdenum chloride, A method of combined administration for preventing the formation of pathological extracellular matrix (ECM) or alleviating or improving a formed pathological extracellular matrix, wherein the agent is one or more selected from organic calcium salts such as molybdenum oxychloride, molybdenum oxide, molybdenum bromide nitrate, molybdenum nitrate, molybdenum sulfide, molybdenum hydroxide, molybdenum oxalate, calcium oxide, calcium silicate, calcium chloride, calcium carbonate, calcium sulfate, calcium nitrate, calcium phosphate, calcium gluconate, calcium lactate, calcium citrate, calcium acetate, and calcium foamate, calcium glycerophosphate, calcium ascorbate, calcium dextran, calcium hydroxyapatite, calcium glucoheptonate, calcium pyruvate, calcium maleate, calcium silicate, and magnesium silicate.

18. A step comprising administering a drug in combination with a composition comprising niclosamide or a pharmaceutically acceptable salt thereof as an active ingredient, A combination administration method for preventing the formation of pathological extracellular matrix (ECM) or for alleviating or improving the formed pathological extracellular matrix.

19. In Claim 18, The above oral administration step is a method of combination administration to prevent the formation of pathological extracellular matrix (ECM) or to alleviate or improve a formed pathological extracellular matrix, wherein niclosamide or a pharmaceutically acceptable salt thereof is administered multiple times.

20. In Claim 19, A combination administration method for preventing the formation of pathological extracellular matrix (ECM) or alleviating or improving formed pathological extracellular matrix, wherein the above-mentioned niclosamide or a pharmaceutically acceptable salt thereof is administered in a range of 10 to 200 mg / kg / day.

21. In Claim 19, The above multiple administration method for preventing the formation of pathological extracellular matrix (ECM) or alleviating or improving a formed pathological extracellular matrix, wherein niclosamide or a pharmaceutically acceptable salt thereof is administered at least twice a day.

22. In Claim 18, A combination administration method for preventing the formation of a pathological extracellular matrix (ECM) or alleviating or improving a formed pathological extracellular matrix, wherein the pathological extracellular matrix (ECM) is induced by at least one of a pathological extracellular matrix caused by ECM modification induced by factors secreted from cells or stromal cells as the disease progresses, and tissue hardening induced by repeated administration of an external drug.

23. In Claim 18, A method of administration for preventing the formation of the above-mentioned pathological extracellular matrix (ECM) or for alleviating or improving the pathological extracellular matrix is ​​a combined administration method for preventing the formation of the pathological extracellular matrix (ECM) or for alleviating or improving the pathological extracellular matrix, wherein the method reduces the expression level of one or more of MMP-9 and VEGF factors.

24. In Claim 18, The composition comprising the above-mentioned niclosamide or a pharmaceutically acceptable salt thereof as an active ingredient further comprises a mineral salt, and The above mineral salts are calcium chloride, calcium sulfate, calcium nitrate, calcium oxide, calcium hydroxide, iron chloride, iron nitrate, iron sulfate, iron hydroxide, potassium chloride, potassium nitrate, potassium hydroxide, sodium chloride, sodium nitrate, sodium hydroxide, zinc chloride, zinc nitrate, zinc sulfate, zinc oxide, zinc hydroxide, magnesium chloride, magnesium nitrate, magnesium sulfate, magnesium oxide, magnesium hydroxide, copper chloride, copper nitrate, copper sulfate, copper oxide, copper hydroxide, manganese chloride, manganese nitrate, manganese sulfate, manganese oxide, manganese hydroxide, chromium chloride, chromium nitrate, chromium sulfate, chromium oxide, chromium hydroxide, cobalt chloride, cobalt nitrate, cobalt sulfate, cobalt oxide, cobalt hydroxide, selenium chloride, selenium nitrate, selenium sulfate, selenium oxide, selenium hydroxide, molybdenum chloride, A method of combined administration for preventing the formation of a pathological extracellular matrix (ECM) or alleviating or improving a formed pathological extracellular matrix, wherein the agent is one or more selected from organic calcium salts such as molybdenum oxychloride, molybdenum oxide, molybdenum bromide nitrate, molybdenum nitrate, molybdenum sulfide, molybdenum hydroxide, molybdenum oxalate, calcium oxide, calcium silicate, calcium chloride, calcium carbonate, calcium sulfate, calcium nitrate, calcium phosphate, calcium gluconate, calcium lactate, calcium citrate, calcium acetate, and calcium foamate, calcium glycerophosphate, calcium ascorbate, calcium dextran, calcium hydroxyapatite, calcium glucoheptonate, calcium pyruvate, calcium maleate, calcium silicate, and magnesium silicate.

25. In Claim 18, A method of co-administration for preventing the formation of pathological extracellular matrix (ECM) or alleviating or improving the formed pathological extracellular matrix, wherein the drug in the above-mentioned co-administration step is one or more selected from anticancer agents, treatments for neurological diseases, anti-inflammatory agents, and immunomodulators.

Images