KR20230081687A - Uses of microsomal triglyceride transfer protein inhibitors for treating fibrotic diseases - Google Patents

Abstract

The present invention relates to the use of a microsomal triglyceride transfer protein (MTP) inhibitor for the treatment of fibrotic diseases, and more particularly, as a novel use of an MTP inhibitor exhibiting an inhibitory effect on collagen accumulation associated with fibrosis, fibrosis It relates to disease prevention or treatment use.
The composition of the present invention containing an MTP inhibitor has a very excellent effect of preventing or treating fibrosis by inhibiting collagen accumulation in cells and tissues caused by inflammatory stimuli, and this effect has a synergistic effect when used in combination with parthenolide or digoxin. Therefore, it can be very useful for preventing or developing a treatment for fibrotic diseases.

Description

Microsomal triglyceride transfer protein inhibitors for treating fibrotic diseases {Uses of microsomal triglyceride transfer protein inhibitors for treating fibrotic diseases}

The present invention relates to the use of a microsomal triglyceride transfer protein (MTP) inhibitor for the treatment of fibrotic diseases, and more particularly, as a novel use of an MTP inhibitor exhibiting an inhibitory effect on collagen accumulation associated with fibrosis, fibrosis It relates to disease prevention or treatment use.

Fibrosis is a disease in which extracellular matrix is abnormally produced, accumulated, and deposited by fibroblasts, and is caused by fibrosis of organs or tissues. Fibrosis is a very fatal disease that causes organ damage. For example, idiopathic pulmonary fibrosis (IPF) occurs as a result of recurrent alveolar epithelial cell damage associated with fibroblast accumulation and myofibroblast differentiation, resulting in irreversible destruction of lung parenchyma tissue and extracellular matrix (ECM). ) is a chronic, progressive, and lethal disease that causes excessive accumulation of However, since there is no effective treatment for fibrosis to date (Fibrogenesis Tissue Repair, 2012, 5(1): 11; N Engl J Med, 2001, 345(7): 517), therapeutic agents that can effectively prevent or treat fibrosis Development is constantly required.

Fibrosis develops as a result of various underlying diseases. Chronic inflammation or tissue damage/remodeling is a typical fibrosis-induced event. Specific disease examples include idiopathic pulmonary fibrosis (IPF), liver fibrosis associated with alcoholic and non-alcoholic liver cirrhosis, renal fibrosis, cardiac fibrosis, and keloid formation resulting from abnormal wound healing [Wynn, T. A. (2004) Nature Reviews. Immunology. 4: 583-594; Friedman, S.L. (2013) Science Translation Medicine. 5(167):1-17]. Additionally, fibrosis is a key pathological feature associated with chronic autoimmune diseases including rheumatoid arthritis, Crohn's disease, systemic lupus erythematosus and scleroderma. Diseases that present a significant unmet medical need include idiopathic pulmonary fibrosis (IPF), scleroderma and nonalcoholic steatohepatitis (NASH) associated liver fibrosis. The increased incidence of NASH-related liver fibrosis is expected to directly parallel that of type 2 diabetes and obesity.

On the other hand, hepatic fibrosis (liver cirrhosis or hepatic fibrosis) or liver cirrhosis (or liver cirrhosis, liver cirrhosis) occurs when liver tissue repeats damage and regeneration, resulting in hepatitis, and progresses from hepatitis to liver fibrosis and from liver fibrosis to cirrhosis. belonging to a disease Liver fibrosis is a fibrosis phenomenon (liver fibrogenesis) that occurs due to the continuous destruction of liver cells and repetitive liver damage. The production of ECM (extracellular matrix) is increased, but the degradation of it is relatively reduced. It progresses to cirrhosis, liver failure or liver cancer. Liver fibrosis is difficult to restore to a normal liver once fibrosis progresses, and it progresses to liver cirrhosis or liver cancer, resulting in a continuously increasing mortality rate. Liver cirrhosis refers to deterioration of liver function by changing normal liver tissue into fibrotic tissue such as regenerative nodules (a phenomenon in which small lumps are formed) due to chronic inflammation. So far, there is no specific treatment for these diseases.

Accordingly, the present inventors have conducted extensive research to develop a therapeutic agent that can effectively and safely treat fibrotic diseases. It was found that it was further improved through combined use, and the present invention was completed.

Accordingly, an object of the present invention is to provide a pharmaceutical composition for preventing or treating fibrotic disease, comprising a microsomal triglyceride transfer protein (MTP) inhibitor as an active ingredient.

In order to achieve the above object of the present invention, the present invention provides a pharmaceutical composition for preventing or treating fibrotic disease comprising a microsomal triglyceride transfer protein (MTP) inhibitor as an active ingredient.

As used herein, the term "treatment" or "treating" refers to the application or administration of a therapeutic agent, that is, a compound of the present invention (alone or in combination with other pharmaceutical agents) to a patient or HBV infection, HBV infection Treatment, cure, alleviation, alleviation, alteration, resolution, amelioration, improvement or outbreak of HBV infection, symptoms of HBV infection, or HBV infection from patients at risk of developing symptoms or HBV infection (e.g., diagnosis or ex vivo application) It is defined as the application or administration of a therapeutic agent to an isolated tissue or cell line for the purpose of influencing the likelihood of becoming infected. Such treatment can be tailored and modified based on knowledge obtained, inter alia, from the field of genomic pharmacology.

As used herein, the term "prevent" or "prevention" means that, if nothing has happened, there is no development of the disorder or disease, or if there has already been the development of the disorder or disease, there is no further development of the disorder or disease. Also contemplated is one's ability to prevent some or all of the symptoms associated with a disorder or disease.

As used herein, the term "patient", "individual" or "subject" refers to a human or a non-human mammal. For example, non-human mammals include livestock and pets such as ovine, bovine, porcine, canine, feline and murine mammals. Preferably the patient, subject or individual is a human.

As used herein, the terms "effective amount", "pharmaceutically effective amount" and "therapeutically effective amount" refer to a non-toxic sufficient amount of an agent to provide a desired biological result. The result may be a reduction and/or attenuation of a signal, symptom or cause of a disease, or any other desired change in a biological system. An appropriate therapeutic amount for any individual can be determined by one skilled in the art using routine experimentation.

As used herein, the term "pharmaceutically acceptable" means that it does not interfere with the biological activity or properties of a compound and is relatively non-toxic, that is, a substance has an undesirable biological effect or any component of a composition it contains. Refers to a substance, such as a carrier or diluent, that can be administered to an individual without interacting in a detrimental way with the component.

As used herein, the term "pharmaceutically acceptable salt" refers to a salt of a compound to be administered prepared from non-toxic acids including pharmaceutically acceptable inorganic acids, organic acids, solvates, hydrates or clathrates thereof. indicate Examples of such inorganic acids are hydrochloric acid, hydrobromic acid, iodic acid, nitric acid, sulfuric acid, phosphoric acid, acetic acid, hexafluorophosphoric acid, citric acid, gluconic acid, benzoic acid, propionic acid, butyric acid, sulfosalicylic acid, maleic acid, lauric acid, malic acid, fumaric acid. , succinic acid, tartaric acid, amsonic acid, palmic acid, p-toluenesulfonic acid and mesylic acid. Suitable organic acids may be selected, for example, from the aliphatic, aromatic, carboxylic and sulfonic classes of organic acids, such as formic acid, acetic acid, propionic acid, succinic acid, camphorsulfonic acid, citric acid, fumaric acid, gluconic acid, isethionic acid, Lactic acid, malic acid, mucous acid, tartaric acid, para-toluenesulfonic acid, glycolic acid, glucuronic acid, maleic acid, furoic acid, glutamic acid, benzoic acid, atranilic acid, salicylic acid, phenylacetic acid, mandelic acid, emphonic acid (pam acid), Examples include methanesulfonic acid, ethanesulfonic acid, pantothenic acid, benzenesulfonic acid (besylate), stearic acid, sulfanilic acid, alginic acid, and galacturonic acid. Moreover, pharmaceutically acceptable salts include, but are not limited to, alkaline earth metal salts (eg calcium or magnesium), alkali metal salts (eg sodium-dependent or potassium) and ammonium salts. includes

As used herein, the term "pharmaceutically acceptable carrier" refers to a liquid or solid filler, stabilizer, A pharmaceutically acceptable substance, composition or carrier such as a dispersing agent, suspending agent, diluent, additive, thickening agent, solvent or encapsulating material. Typically, these components are carried or transported from one organ or part of the body to another organ or part of the body. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formula, including compounds useful within the present invention, and not injurious to the patient. Some examples of materials that can serve as pharmaceutically acceptable carriers include: sugars such as lactose, glucose and sucrose used in pharmaceutical formulations; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethyl cellulose, ethylcellulose and cellulose acetate; tragacanth powder; malt; gelatin; talc; additives such as cocoa butter and suppository wax; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; glycols such as propylene glycol; polyols such as glycerin, sorbitol, mannitol and polyethylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffers such as magnesium hydroxide and aluminum hydroxide; Surfactants; alginic acid; pyrogen-free water; isotonic solution; Ringer's solution; ethyl alcohol; phosphate buffer solution; and other non-toxic compatible substances. In addition, as used in the present invention, "pharmaceutically acceptable carrier" is compatible with the activity of compounds useful within the present invention, and is physiologically acceptable to patients in part or in whole coating agent, antiviral and antibacterial agent, and absorption delaying agent. Include etc. Supplementary active compounds may also be incorporated into the compositions. Further "pharmaceutically acceptable carriers" may further include pharmaceutically acceptable salts of compounds useful within the present invention. Other additional ingredients that may be included in the pharmaceutical compositions used in the practice of the present invention are known in the art and are described, for example, in Remington's Pharmaceutical Sciences (Genaro, Ed., Mack Publishing Co. , 1985, Easton, PA).

As used herein, the term "composition" or "pharmaceutical composition" refers to a mixture of at least one compound useful within the present invention with a pharmaceutically acceptable carrier. A pharmaceutical composition facilitates administration of the compound to a patient or subject. A variety of techniques exist in the art for administering compounds including, but not limited to, intravenous, oral, aerosol, parenteral, ocular, pulmonary and topical administration.

As used herein, the terms "combination therapy" or "in combination" refer to the administration of two or more therapeutic agents to treat the conditions or disorders described herein (ie, metabolic diseases and fibrotic diseases). Such administration includes co-administration of these therapeutic agents in a substantially simultaneous manner, such as in a single capsule having a fixed proportion of the active ingredients. Alternatively, such administration includes co-administration in multiple or separate containers (eg, capsules, powders, and liquids) for each active ingredient. Powders and/or liquids may be reconstituted or diluted to the desired dosage prior to administration. In addition, such administration includes sequential use of each type of therapeutic agent at about the same time or at different times. In either case, the treatment regimen will provide beneficial effects of the drug combination in the treatment of the conditions or disorders described herein.

Combination therapy can provide a "synergistic effect" and can prove to be "synergistic." That is, the effect achieved when the active ingredients are used together is greater than the sum of the effects produced by using the compounds individually. A synergistic effect occurs when the active ingredients are (1) co-formulated and administered or delivered simultaneously in a combined unit dosage form; (2) when delivered alternately or in parallel as separate dosage forms; or (3) when delivered by some other initiative. When delivered in alternating therapy, a synergistic effect can be obtained when the compounds are administered or delivered sequentially, eg by different injections in separate syringes. Generally, during alternation therapy, effective doses of each active ingredient are administered sequentially, i.e. consecutively, whereas in combination therapy, effective dosages of two or more active ingredients are administered together. Synergistic effect as used herein refers to the action of two therapeutic agents to produce an effect greater than the simple sum of the effects of each drug administered alone. Synergistic effects are described, for example, by the Sigmoid-Emax equation (Holford, N. H. G. and Scheiner, L. B., Clin. Pharmacokinet. 6: 429-453 (1981)), the Loewe additive equation (Loewe, S. and Muischnek, H., Arch. Exp. Pathol Pharmacol. 114: 313-326 (1926)), and the median effect equation (Chou, T. C. and Talalay, P., Adv. Enzyme Regul. 22: 27-55 (1984)). can be calculated. Each of the equations mentioned above can be applied to experimental data to generate corresponding graphs that help evaluate the effects of drug combinations. Corresponding graphs related to the above-mentioned equations are concentration-effect curves, isobologram curves, and combination index curves, respectively.

Hereinafter, the present invention will be described in detail.

The present invention provides a pharmaceutical composition for preventing or treating fibrotic disease comprising a microsomal triglyceride transfer protein (MTP) inhibitor as an active ingredient.

MTP is the protein encoded by the MTTP gene in humans, which completes the heterodimeric microsomal triaglyceride transporter protein, which plays a central role in lipoprotein assembly. MTP is a lipid transport protein required for the assembly and secretion of very-low-density lipoproteins (LDL) in the liver and chylomicrons in the intestine. In the present invention, the MTP inhibitor may include those that inhibit the expression of a gene encoding an MTP protein or the activity of a protein. For example, antisense nucleotides, aptamers, small interfering RNA (siRNA), short hairpin RNA (shRNA), micro RNA (miRNA) or RNA interference (RNAi) against MTTP mRNA encoding the MTP protein, and the like, and It may include, but is not limited to, natural products, organic compounds, poorly differentiated compounds, antibodies, proteins, etc. capable of inhibiting expression and/or activity.

According to one embodiment of the present invention, it was confirmed that the MTP inhibitor has a very excellent activity of inhibiting collagen accumulation by inflammatory factors in human lung fibroblasts, and can exhibit an effect of preventing or treating fibrotic disease in each tissue in the body.

In the present invention, the fibrotic disease is also called fibrosis, and is a chronic and progressive disease characterized by excessive accumulation of extracellular matrix (ECM) leading to hardening and/or scarring of related tissues. It develops through complex interactions of cells, extracellular matrix, cytokines, and growth factors. Other cell types include resident mesenchymal cells (fibroblasts and myofibroblasts) and ECM-producing cells derived from epithelial and endothelial cells (through a process called epithelial- and endothelial-mesenchymal transition), and local or bone marrow-derived stem cells (fibroblasts). ), etc. Fibroblasts have long been considered as the major cell type involved in normal wound healing and as important effector cells in fibrogenesis. They are highly synthetic to collagen and other ECM components, and are also characterized by de novo expression of α-smooth muscle actin (α-SMA) (reviewed in Scotton C.J. and Chambers R.C., 2007). The presence of myofibroblasts in fibrotic lesions in animal models of fibrosis correlates with the onset of active fibrosis, and the persistence and localization of fibrotic lesions in humans is associated with disease progression (Kuhn C. and McDonald J.A., 1991, and Zhang et al., 1994). Myofibroblasts also display an enhanced migration phenotype (Suganuma et al. 1995) and are capable of releasing multiple mediators of profibrosis.

In the present invention, the type of fibrotic disease is not particularly limited, and is caused by chronic inflammation or tissue damage/remodeling, etc., and is a progressive disease that leads to hardening and/or scarring of related tissues due to excessive accumulation of extracellular matrix (ECM). Any disease can be included. Non-limiting examples of the fibrotic disease may include pulmonary fibrosis, liver fibrosis, skin fibrosis, renal fibrosis, pancreatic fibrosis, systemic sclerosis and cardiac fibrosis.

The lung fibrosis is a serious medical condition involving scarring of lung tissue. These symptoms occur when the alveoli and interstitial tissue of the lungs become inflamed and develop scars in the tissue in an attempt to repair itself. Pulmonary fibrosis involves progressive exchange of fibrotic tissue (fibrous scar) with normal lung parenchyma.

Pulmonary fibrosis is characterized by chronic inflammatory processes (sarcomatosis, Wegener's granulomatous disease), infections, environmental agents (exposure to asbestos, silica, certain gases), exposure to ionizing radiation (e.g. radiation therapy to treat tumors in the chest), and chronic conditions. (lupus), and certain medications (eg amiodarone, bleomycin, pingyangmycin, busulfan, methotrexate, and nitrofurantoin).

In a condition known as hypersensitivity pneumonitis, fibrosis of the lungs may develop after a high immune response to inhaled organic dust or industrial chemicals. These symptoms mostly result from inhalation of dust contaminated with bacteria, fungi, or animal products.

The liver fibrosis or hepatic fibrosis is a process of excessive accumulation of extracellular matrix proteins (including collagen) and subsequent scar formation that occurs in most chronic liver diseases. Liver fibrosis that progresses over time results in cirrhosis. Cirrhosis is the final stage of chronic liver disease and is generally irreversible with poor long-term prognosis. At an advanced stage, the only option is a liver transplant. The risk of liver cancer is significantly increased with cirrhosis, and cirrhosis can be seen as a precancerous condition (hepatocellular carcinoma). In fact, cirrhosis and liver cancer are among the top 10 causes of death worldwide. Thus, there is a need for effective treatment of liver fibrosis and subsequent cirrhosis. Unfortunately, there are few treatment options available, and most treatment consists of addressing the causes and/or symptoms of cirrhosis.

A limited number of clinical trials are ongoing with candidate drugs that specifically address the inhibition or retardation of fibrosis in the liver. However, these trials target specific liver diseases, such as non-alcoholic steatohepatitis (NASH).

The skin fibrosis or epidermal fibrosis is excessive scarring of the skin and results from a pathological wound healing response. There are a wide range of fibrotic skin diseases: scleroderma, nephrogenic dermatosis, mixed connective tissue disease, sclerosing myxedema, sclerotic hydrocele, and eosinophilic fasciitis. Exposure to chemical or physical agents (mechanical trauma, burns) is also a potential cause of fibrotic skin disease. Skin fibrosis can be driven by immune, autoimmune and inflammatory mechanisms. The balance of collagen production and degradation in fibroblasts plays an important role in the pathophysiological process of skin fibrosis. Certain cytokines such as transforming growth factor-β (TGB-β) and interleukin-4 (IL-4) promote wound healing and fibrosis, while interferon-γ (IFN-γ) and tumor necrosis factor-α ( Others, such as TNF-α), are anti-fibrotic. Fibroblasts in normal skin are quiescent. They control the amount of connective tissue proteins and have low proliferative activity. After skin injury, these cells are activated, ie they express α-smooth muscle actin (α-SMA) and synthesize large amounts of connective tissue proteins. Activated cells are often called myofibroblasts.

scleroderma refers to skin fibrosis; Sclera = hard and dermal skin. However, dermal fibrosis can have a significant impact on health outcomes, particularly when it is part of systemic scleroderma. The latter refers to connective tissue diseases of autoimmune etiology. Restricted cutaneous scleroderma is limited to the skin of the face and feet, whereas diffuse cutaneous scleroderma covers more skin and may progress to the visceral organs.

The renal fibrosis means excessive proliferation of cells, hardening of tissues and scars. Renal fibrosis can result from renal failure and catheterization, eg dialysis following peritoneal and vascular access fibrosis. Renal fibrosis can also result from nephropathy such as glomerular disease (eg glomerulosclerosis, glomerulonephritis), chronic renal failure, acute renal failure, end-stage renal disease and renal failure. Regardless of etiology, all patients with chronic kidney disease exhibit a gradual decline in renal function over time. Fibrosis, so-called scarring, is a major cause of the pathophysiology. Fibrosis involves excessive accumulation of extracellular matrix (composed primarily of collagen), usually resulting in loss of function when normal tissue is replaced by scar tissue. This process is often irreversible and essentially leads to end-stage renal failure, a condition requiring lifelong dialysis or kidney transplantation.

The cardiac fibrosis, a hallmark of heart disease, is thought to contribute to sudden cardiac death, ventricular arrhythmias, left ventricular (LV) dysfunction, and cardiac disorders. Cardiac fibrosis is characterized by disproportionate accumulation of fibrotic collagen, inflammation, increased workload, hypertrophy, and stimulation by a number of hormones, cytokines, and growth factors that occur after cardiomyocyte death.

Cardiac fibrosis can also refer to abnormal thickening of heart valves due to disproportionate proliferation of cardiac fibroblasts, but more generally refers to proliferation of fibroblasts in the heart muscle. Fibrous cells normally secrete collagen and function to provide structural support for the heart. When overly activated, this process causes thickening and fibrosis of the valves, at which time white tissue not only builds the tricuspid valve, but also occurs in the pulmonary valve. Concentration and loss of flexibility can eventually lead to valve failure and right-sided heart failure.

Chronic pancreatitis (CP), which causes pancreatic fibrosis, is a progressive inflammatory disease of the pancreas characterized by irreversible morphological changes and progressive fibrotic replacement of the gland. In parenchymal fibrosis, loss of exocrine and endocrine function occurs. The main symptoms of CP are abdominal pain and indigestion. Extremely, the pancreas may be enlarged or atrophied with or without cysts or calcifications or tumors. The tube may be dilated, irregular or constricted. The major pathological features include irregular and non-uniform loss of lobular tissue, chronic inflammation, ductal changes and fibrosis. Gene mutations (including but not limited to cystic fibrosis, cationic trypsinogen gene, CFTR gene mutation in idiopathic acute and chronic pancreatitis, pancreatic secretory trypsin inhibitor gene, chymotrypsinogen C gene, calcium sensing receptor gene, α- 1 including antitrypsin deficiency), metabolic (alcohol, tobacco smoking, hypercalcemia, hyperlipidemia, chronic renal failure), environmental factors (micronutrient deficiencies (nutritional factors such as zinc, copper and selenium; also post-irradiation exposure), obstructive ( tumor), ischemic (vascular disease)) and autoimmune diseases, or is the final manifestation of a complex pathogenic mechanism associated with primary sclerosing bile duct flames, Sjogren's syndrome, primary biliary disorders and type 1 diabetes mellitus.

In such various fibrosis, the composition containing the non-nucleoside reverse transcriptase inhibitor according to the present invention suppresses the fibrotic reaction by reducing the amount of procollagen alpha 1 in each tissue or cell, and this effect is (B-cell lymphoma 2) inhibitors, DPP-4 (Dipeptidyl peptidase-4) inhibitors, and niclosamide (Niclosamide) when used in combination with one or more drugs can show synergistic effects.

In one aspect of the present invention, the composition may further include at least one selected from the group consisting of parthenolide, digoxin, and pharmaceutically acceptable salts thereof. That is, an MTP inhibitor and parthenolide or digoxin can show a synergistic effect in the treatment of fibrotic disease as a combination therapy or as a combination agent.

In a preferred aspect of the present invention, lomitapide as an MTP inhibitor; and parthenolide or digoxin, the combination therapy or combination preparation may show a synergistic effect in the treatment of fibrotic disease.

In this case, each component included in the combination preparation may be formulated for use separately, simultaneously or sequentially. That is, in the case of sequential administration, the combination preparation may be administered in two or more administrations. Concomitant administration can include co-administration using separate formulations or a single pharmaceutical formulation, as well as sequential administration in either order.

In the present invention, when the MTP inhibitor and parthenolide are used as a combination therapy or combination drug, these combination drugs are used in a molar ratio of 1:0.01 to 1000, preferably in a molar ratio of 1:1 to 500, more preferably It may be administered in combination at a molar ratio of 1:50 to 500, more preferably at a molar ratio of 1:100 to 500, and most preferably at a molar ratio of 1:150 to 350.

In the present invention, when the MTP inhibitor and digoxin are used as a combination therapy or combination drug, these combination drugs are used in a molar ratio of 1:0.1 to 100, preferably in a molar ratio of 1:0.1 to 50, and more preferably 1 : It may be co-administered at a molar ratio of 0.5 to 20, most preferably at a molar ratio of 1:1 to 10.

The present invention also relates to lomitapide or a pharmaceutically acceptable salt thereof; And parthenolide (Parthenolide), digoxin (Dogoxin) and provides a pharmaceutical composition for preventing or treating fibrotic disease comprising at least one selected from the group consisting of pharmaceutically acceptable salts thereof as an active ingredient.

The pharmaceutical composition according to the present invention may contain the MTP inhibitor alone or may be formulated in a suitable form together with a pharmaceutically acceptable carrier, and may further contain an excipient or diluent. In the above, 'pharmaceutically acceptable' refers to a non-toxic composition that is physiologically acceptable and does not cause allergic reactions such as gastrointestinal disorders, dizziness, etc., or similar reactions when administered to humans.

A pharmaceutically acceptable carrier may further include, for example, a carrier for oral administration or a carrier for parenteral administration.

Carriers for oral administration may include lactose, starch, cellulose derivatives, magnesium stearate, stearic acid and the like.

In addition, various drug delivery materials used for oral administration may be included. In addition, the carrier for parenteral administration may include water, suitable oil, saline, aqueous glucose and glycol, and the like, and may further include a stabilizer and a preservative. Suitable stabilizers include antioxidants such as sodium bisulfite, sodium sulfite or ascorbic acid. Suitable preservatives include benzalkonium chloride, methyl- or propyl-paraben and chlorobutanol.

The pharmaceutical composition of the present invention may further include a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifier, a suspending agent, and the like in addition to the above components. Other pharmaceutically acceptable carriers and formulations may be referenced as described in the following literature (Remington's Pharmaceutical Sciences, 19th ed., Mack Publishing Company, Easton, PA, 1995).

The composition of the present invention can be administered to mammals including humans by any method. For example, it can be administered orally or parenterally. Parenteral administration methods include, but are not limited to, intravenous, intramuscular, intraarterial, intramedullary, intrathecal, intracardiac, transdermal, subcutaneous, intraperitoneal, intranasal, intestinal, topical, sublingual or rectal administration can be

The pharmaceutical composition of the present invention may be formulated into a formulation for oral administration or parenteral administration according to the administration route as described above.

In the case of preparations for oral administration, the composition of the present invention may be formulated into powders, granules, tablets, pills, dragees, capsules, solutions, gels, syrups, slurries, suspensions, etc. using a method known in the art. can For example, preparations for oral use may be obtained by combining the active ingredient with a solid excipient, which is then milled and, after adding suitable auxiliaries, processed into a mixture of granules to obtain tablets or dragees. Examples of suitable excipients include sugars including lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol and maltitol, starches including corn starch, wheat starch, rice starch and potato starch, cellulose, Celluloses including methyl cellulose, sodium carboxymethylcellulose and hydroxypropylmethyl-cellulose, and the like, fillers such as gelatin, polyvinylpyrrolidone, and the like may be included. In addition, cross-linked polyvinylpyrrolidone, agar, alginic acid or sodium alginate may be added as a disintegrant, if desired.

Furthermore, the pharmaceutical composition of the present invention may further include an anticoagulant, a lubricant, a wetting agent, a flavoring agent, an emulsifier, and a preservative.

In the case of preparations for parenteral administration, they may be formulated in the form of injections, creams, lotions, external ointments, oils, moisturizers, gels, aerosols, and nasal inhalants by methods known in the art. These formulations are described in Remington's Pharmaceutical Science, 19th ed., Mack Publishing Company, Easton, PA, 1995, which is a generally known formula for all pharmaceutical chemistry.

The total effective amount of the composition of the present invention can be administered to the patient in a single dose or by a fractionated treatment protocol in which multiple doses are administered over a long period of time. The pharmaceutical composition of the present invention may vary the content of the active ingredient according to the severity of the disease. Preferably, the preferred total dose of the pharmaceutical composition of the present invention may be about 0.01 to 10,000 mg, most preferably 0.1 to 500 mg per patient body weight per day. However, the dosage of the pharmaceutical composition is determined by considering various factors such as the formulation method, administration route, and number of treatments as well as the patient's age, weight, health condition, sex, severity of disease, diet and excretion rate, etc. Therefore, considering this point, those skilled in the art will be able to determine an appropriate effective dosage of the composition of the present invention. The pharmaceutical composition according to the present invention is not particularly limited in its formulation, administration route and administration method as long as it exhibits the effects of the present invention.

In such various fibrosis, the composition containing the MTP inhibitor according to the present invention suppresses the fibrotic reaction by reducing the accumulation of collagen by inflammatory stimulation in each tissue or cell, and this effect is synergistic when used in combination with parthenolide. effect may appear.

The composition of the present invention containing an MTP inhibitor has a very excellent effect of preventing or treating fibrosis by inhibiting collagen accumulation in cells and tissues caused by inflammatory stimuli, and this effect has a synergistic effect when used in combination with parthenolide or digoxin. Therefore, it can be very useful for preventing or developing a treatment for fibrotic diseases.

1 is a result of evaluating collagen production inhibitory ability by Sircol assay after a single treatment of lomitapide, parthenolide, digoxin, and a control drug (Nint) on fibrosis-induced Primary Human Pulmonary Fibroblasts, respectively.
Figure 2 is a result (Kit) of evaluating the anti-fibrotic efficacy of lomitapide in a pulmonary fibrosis animal model (BLM IPF mouse model) by hydroxyproline assay.
Figure 3 is the result (LC-MS) of evaluating the anti-fibrotic efficacy of lomitapide in a pulmonary fibrosis animal model (BLM IPF mouse model) by hydroxyproline assay (T1: lomitapide 10mg/kg, T2: lomitapide 50mg /kg).
Figure 4 is a result of evaluating the anti-fibrosis efficacy according to lomitapide, digoxin, or a combination treatment thereof by expression changes of fibrosis-related genes.
5 is a result of evaluating the anti-fibrotic efficacy of lomitapide, digoxin or a combination thereof through changes in collagen secretion in cells.
Figure 6 is a result of evaluating the anti-fibrotic efficacy according to lomitapide, parthenolide or a combination thereof by the expression change of the fibrosis-related gene COL1A1.
7 is a result of evaluating the anti-fibrotic efficacy according to lomitapide, parthenolide or a combination treatment thereof by the expression change of a-SMA, a fibrosis-related gene.
8 is a result of evaluating the anti-fibrotic efficacy of lomitapide, parthenolide, or a combination thereof through changes in collagen secretion in cells.

Hereinafter, the present invention will be described in detail by the following examples. However, the following examples are only for illustrating the present invention, and the present invention is not limited thereto.

Example 1: Evaluation of collagen accumulation inhibitory ability

We selected lomitapide, parthenolide, and digoxin as single-agent drugs predicted to show fibrogenic inhibition by AI-based analysis of patient genome data published in papers, etc. The ability to inhibit collagen formation was confirmed in vitro.

Experiments were performed using the Sircol Collagen Assay kit according to the manufacturer's instructions. As a control drug, Nintedanib (Nint) 1uM, a marketed IPF drug, was used. Nintedanib was treated at a high concentration of 1uM with reference to the paper, and lomitapide, parthenolide and digoxin were also treated at a concentration of 1uM.

Specifically, experiments were performed with Primary Human Pulmonary Fibroblasts isolated from human lung tissue (purchased by Promo Cell). Cells were seeded in 6 wells at a density of 1 x 10 ⁵ , and fibrosis was induced by treatment with the recombinant TGF-beta at a concentration of 20 ng/mL for 44 hr. Each drug was pre-treated 4 hours before TGF-beta stimulation, harvested and analyzed 48 hours after drug treatment. The results for this are shown in Figure 1.

Example 2: In vivo fibrosis inhibitory activity evaluation

In the BLM IPF mouse model, the anti-fibrotic efficacy of lomitapide was confirmed at two doses. The specific experimental conditions are as follows:

Experimental animal: male mouse (Specific Pathogen Free; SPF) C57BL/6NCrlOri

Pulmonary fibrosis inducer: Prepared by dissolving Bleomycin sulfate (BLM) in crinkle irrigation agent (sterilized physiological saline) as an excipient

Composition of test substance and positive control vehicle: 5% DMSO + 5% Cremophor EL + 90% Saline

Preparation of test substance: preparation on the day of administration

BLM and drug administration

Administration method

Lung fibrosis inducer and excipient: BLM was administered single intra-airway to all animals on Day 1. Before intratracheal instillation, anesthesia was performed for about 1 to 5 minutes using isoflurane inhalation.

Test substance and excipient: The test substance and test substance excipient were prepared on the day of administration and repeatedly orally administered on Days 1 to 21.

Positive control substance (pirfenidone): The positive control substance was orally administered repeatedly on Days 1-21.

BioVision's Hydroxyproline assay kit was used and the procedure was performed with reference to the attached protocol.

Specifically, it was homogenized with 50 ul of water per 10 mg of lung tissue, and after adding the same amount of HCl, it was modified to hydrolyze conditions at 120 ° C for 5 hours, and unexpectedly, all the kit protocols were performed. OD values were measured using a BioTek SYNERGY Mx instrument.

The results for this are shown in Figure 2.

As shown in Figure 2, it was confirmed that the lomitapide-administered group showed significantly better antifibrotic efficacy than the positive control group.

In order to confirm the results in FIG. 2 once again, the concentration of hydroxyproline was measured by LC-MS.

Specifically, the lung tissue was homogenized, normalized with the amount of tertiary distilled water, and then hydrolyzed at 120 degrees for 24 hours after adding HCl. It was analyzed by LC-MS.

The results for this are shown in Figure 3.

As shown in Figure 3, it was confirmed that the lomitapide-administered group showed excellent antifibrotic efficacy equivalent to that of the positive control group.

Example 3: In vitro fibrosis inhibitory ability (1)

When digoxin (Digoxin), which was confirmed to exhibit antifibrotic efficacy in Example 1, was treated in combination with lomitapide, it was attempted to determine whether a synergistic effect appeared.

Specifically, the group treated with 19.89 nM of lomitapide and 100 nM of digoxin, respectively, and the combined treatment with 19.89 nM of lomitapide and 100 nM of digoxin were confirmed for fibrosis-related gene expression changes. Primary Human Pulmonary Fibroblasts isolated from human lung tissue (purchased by Promo Cell) were used.

Cells were seeded in 6 wells at a density of 1 x 10 ⁶ , and cell fibrosis was induced by treatment with recombinant TGF-beta at a concentration of 20 ng/mL for 20 hr. Each drug was pre-treated 4 hours before TGF-beta stimulation, and harvested and analyzed 24 hours after drug treatment.

The results for this are shown in FIG. 4 .

As shown in Figure 4, it was confirmed that the lomitapide and digoxin combined treatment group exhibits significantly improved antifibrotic activity compared to each single treatment group.

The effect of the combined treatment of lomitapide and digoxin was confirmed again through sircol collagen assay, and it was confirmed whether the amount of soluble collagen secreted from cells actually decreased.

Specifically, the results of the group treated with lomitapide or digoxin at a concentration of 1uM as a single agent and the group administered with 0.5uM each of them were compared. Primary Human Pulmonary Fibroblasts isolated from human lung tissue (purchased by Promo Cell) were used.

Cells were seeded in 6 wells at a density of 2 x 10 ⁶ , and cell fibrosis was induced by treatment with recombinant TGF-beta at a concentration of 20 ng/mL for 44 hr. Each drug was pre-treated 4 hours before TGF-beta stimulation, and harvested and analyzed 48 hours after drug treatment. The included protocol was tested using the Sircol Collagen Assay kit, and Nintedanib 1uM, a marketed IPF drug, was used as a control drug.

The results for this are shown in FIG. 5 .

As shown in FIG. 5, the amount of collagen secretion was reduced even in the lomitapide and digoxin alone-treated group, and in the combination-treated group, a significantly improved decrease in collagen secretion was confirmed compared to the respective drug-only treated group. That is, it was determined that each of these drugs exhibited a synergistic effect by the combined treatment.

Example 4: In vitro fibrosis inhibitory ability (2)

When lomitapide at Cmax concentration (19.89nM) and parthenolide at Cmax concentration (3uM) were administered alone or in combination, concentration-dependent efficacy was confirmed at the mRNA level.

Lomitapide was treated at ½ Cmax and Cmax concentrations, respectively, and co-administered with parthenolide at Cmax concentration.

Specifically, experiments were performed with Primary Human Pulmonary Fibroblasts isolated from human lung tissue (purchased by Promo Cell). Cells were seeded in 12 wells at a density of 5 x 10 ⁵ , and fibrosis was induced by treatment with the recombinant TGF-beta at a concentration of 20 ng/mL for 18 hr. Each drug was pre-treated 4 hours before TGF-beta stimulation, harvested 24 hours after drug treatment, and analyzed the change in the expression level of two marker genes related to fibrosis, a-SMA and COL1A1, and the change in collagen accumulation (sircol collagen assay) did

The results for this are shown in Figures 6 to 8.

As can be seen in FIGS. 6 to 8, the lomitapide and parthenolide combined treatment group showed a value of less than 1 in the drug combination index calculated using the bliss independence model, resulting in a synergistic effect compared to each single treatment group It was confirmed that there is

The composition of the present invention containing an MTP inhibitor has a very excellent effect of preventing or treating fibrosis by inhibiting collagen accumulation in cells and tissues caused by inflammatory stimuli, and this effect has a synergistic effect when used in combination with parthenolide or digoxin. Since it can appear, it can be very useful for preventing or developing a treatment for fibrotic disease, so industrial applicability is very high.

Claims (8)

A pharmaceutical composition for preventing or treating fibrotic disease comprising a microsomal triglyceride transfer protein (MTP) inhibitor as an active ingredient.
The method of claim 1, wherein the MTP inhibitor is lomitapide, implitapide, dilotapide, mitratapide, SLx-4090, CP-346086, CP-346 , CP-741952, JTT-130, R-256918 and a pharmaceutical composition characterized in that selected from the group consisting of pharmaceutically acceptable salts thereof.
The pharmaceutical composition according to claim 1, wherein the fibrotic disease is selected from the group consisting of pulmonary fibrosis, liver fibrosis, skin fibrosis, kidney fibrosis, pancreatic fibrosis, systemic sclerosis and cardiac fibrosis.
The pharmaceutical composition according to claim 1, further comprising at least one selected from the group consisting of parthenolide, digoxin, and pharmaceutically acceptable salts thereof.
The method of claim 4, wherein the MTP inhibitor; And digoxin (Dogoxin) and at least one selected from the group consisting of pharmaceutically acceptable salts thereof are simultaneously, individually or sequentially administered in combination, a pharmaceutical composition characterized in that.
The pharmaceutical composition according to claim 4, wherein the MTP inhibitor and parthenolide are co-administered in a molar ratio of 1:0.01 to 1000.
The pharmaceutical composition according to claim 4, wherein the MTP inhibitor and digoxin are co-administered in a molar ratio of 1:0.1 to 100.
lomitapide or a pharmaceutically acceptable salt thereof; And Parthenolide (Parthenolide), digoxin (Dogoxin) and a pharmaceutical composition for preventing or treating fibrotic disease comprising at least one selected from the group consisting of pharmaceutically acceptable salts thereof as an active ingredient.

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