Pharmaceuticals to prevent or treat fibrosis

Abstract

The pharmaceutical product under this invention is used with specific therapeutic regimens and dosages, and may be useful in preventing or treating fibrosis.

Description

Pharmaceutical composition for the prevention or treatment of fibrosis

[0001] The present invention provides a pharmaceutical composition for preventing or treating fibrosis.

[0002] Fibrosis refers to the hardening of a portion of an organ for any reason. Pulmonary fibrosis and liver fibrosis are representative examples. Pulmonary fibrosis is most often caused by radiation exposure or pulmonary edema, but some people can develop pulmonary fibrosis. There is currently no cure for fibrosis, and treatment options are still being developed and researched.Types of fibrosis include interstitial lung disease (ILD), scleroderma, keloid, hypertrophic scar, non-alcoholic fatty liver disease, primary sclerosing cholangitis (PSC), primary biliary cholangitis (PBC), diabetic retinopathy, age-related macular degeneration (AMD), hypertrophic cardiomyopathy, myocardial infarction, muscular dystrophy, diabetic kidney disease, focal segmental glomerulosclerosis (FSGS), inflammatory bowel disease, IBD) and interstitial lung diseases include idiopathic pulmonary fibrosis (IPF), systemic sclerosis associated interstitial lung disease (SSc-ILD), and chronic fibrosing interstitial lung diseases with a progressive phenotype (PF-ILD).

[0003]

[0004] Idiopathic pulmonary fibrosis (IPF) is a rare, chronic, progressive interstitial lung disease known for its poor prognosis and lack of proven treatment. To date, no clear cause has been identified, and the five-year survival rate after diagnosis is only 43%, and the ten-year survival rate is approximately 15%. Despite extensive research, no treatment has yet been shown to improve survival. Considering that other interstitial lung diseases, such as nonspecific interstitial pneumonia (NSIP) and idiopathic organizing pneumonia (COP), have relatively good prognoses with appropriate treatment, IPF can be considered one of the most unfavorable interstitial lung diseases. The most common causes of death are respiratory failure (39%) and cardiac disease (27%), with other causes including lung cancer, pulmonary embolism, and pneumonia. The prognosis is worse in older age, male gender, those with poor lung function at diagnosis, or those with a high number of fibroblastic foci in biopsies.

[0005]

[0006] Treatment for idiopathic pulmonary fibrosis (IPF) is similar to that for nonspecific interstitial pneumonia (NSIP), with steroids and cytotoxic drugs sometimes used. Recently, antifibrotic agents have become the mainstay of treatment, with various trials underway. Currently, pirfenidone and nintedanib are approved for IPF. These drugs do not provide a cure, but rather delay pulmonary fibrosis and alleviate symptoms. Therefore, there is a need for more effective medications that can improve patients' quality of life.

[0007]

[0008] Systemic sclerosis-associated interstitial lung disease (SSc-ILD) is a condition in which patients with systemic sclerosis (SSc) develop interstitial lung disease (ILD) as a complication, and lung function decline is the leading cause of death in SSc. Nintedanib and tocilizumab, which have been shown to reduce lung function decline, are currently approved treatments for this condition in the United States. However, as with idiopathic pulmonary fibrosis, there is a need for more effective medications that can improve patients' quality of life.

[0009]

[0010] Chronic fibrosing interstitial lung diseases with a progressive phenotype (PF-ILD) refers to various progressive fibrosing interstitial lung diseases excluding idiopathic pulmonary fibrosis, including autoimmune interstitial lung disease and idiopathic interstitial pneumonia. Nintedanib was approved for treatment in the United States after being confirmed to be effective in reducing lung function decline in patients with various fibrosing lung diseases. Since fibrosing interstitial lung diseases can present with a progressive phenotype such as pulmonary fibrosis, lung function decline, and worsening quality of life, regardless of classification and underlying disease, if a drug is proven to be effective in reducing lung function decline in a specific interstitial lung disease, it can be expected to be effective in reducing lung function decline in other interstitial lung diseases as well.

[0011]

[0012] Meanwhile, prolyl-tRNA synthetase (PRS) is a member of the aminoacyl-tRNA synthetase (ARS) family of enzymes that activates amino acids for protein synthesis. Specifically, ARS forms aminoacyl adenylate (AA-AMP) and then transfers the activated amino acid to the 3-terminal of the corresponding tRNA (translational function). Because ARS plays a key role in protein synthesis, ARS inhibition inhibits the growth and development of all cells. Accordingly, ARS is recognized as a promising target for antibiotics and therapeutic agents that require inhibition of cell overexpression (Nature, 2013, 494: 121-125).

[0013]

[0014] PRS exists and functions as a multisynthetase complex (MSC) in the form of EPRS (Glutamyl-Prolyl-tRNA Synthetase). In particular, among various MSCs, EPRS functions as a translational silencer that suppresses the production of vascular endothelial growth factor A (VEGF A), a key factor in angiogenesis, and has also been reported to be closely related to various solid cancers (Nat. Rev. Cancer, 2011, 11, 708-718).

[0015]

[0016] Accordingly, the inventors of the present invention have studied a method for preventing or treating fibrosis and, as a result, have confirmed that effective prevention or treatment of fibrosis is possible when a specific PRS inhibitor, which will be described later, is used at a specific therapeutic dosage, thereby completing the present invention.

[0017] The present invention provides a pharmaceutical composition that can be usefully used for the prevention or treatment of fibrosis.

[0018] To solve the above problem, the present invention provides a pharmaceutical composition for preventing or treating fibrosis as follows:

[0019] A pharmaceutical composition for the prevention or treatment of fibrosis, comprising a compound represented by the following chemical formula 1, or a pharmaceutically acceptable salt thereof, for administration of 100 to 150 mg twice daily (BID):

[0020] [Chemical Formula 1]

[0021] .

[0022]

[0023] The compound represented by the above chemical formula 1 or a pharmaceutically acceptable salt thereof is a compound described in Korean Patent Registration No. 10-2084772, specifically, a substance described as Example 40 of the specification. The substance can be used as a PRS inhibitor for the prevention or treatment of fibrosis.

[0024]

[0025] In particular, as in the present invention, when administered twice daily (BID) at 100 to 150 mg, effective prevention or treatment of fibrosis is possible. The above dosage and administration is a dosage that can increase the preventive or therapeutic effect of the compound represented by the above chemical formula 1, or a pharmaceutically acceptable salt thereof, while at the same time maximally preventing potential risks. In addition, by the above dosage and administration, the area under the plasma drug concentration-time curve (AUC) after administration of the compound represented by the above chemical formula 1, or a pharmaceutically acceptable salt thereof inf ), that is, in vivo exposure may produce optimal results in the prevention or treatment of fibrosis.

[0026]

[0027] Meanwhile, the compound represented by the above chemical formula 1 can be used in the form of a pharmaceutically acceptable salt, and as a salt, an acid addition salt formed by a pharmaceutically acceptable free acid is useful. Inorganic acids and organic acids can be used. Inorganic acids that can be used include hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, etc., and organic acids that can be used include citric acid, acetic acid, lactic acid, maleic acid, gluconic acid, methanesulfonic acid, succinic acid, 4-toluenesulfonic acid, glutamic acid, or aspartic acid. Preferably, the pharmaceutically acceptable salt of the compound represented by the above chemical formula 1 is a hydrochloride salt.

[0028]

[0029] In addition, the compound represented by the above chemical formula 1 can be prepared in a crystalline or amorphous form, and when prepared in a crystalline form, can be optionally hydrated or solvated. The present invention may include not only stoichiometric hydrates of the compound represented by the above chemical formula 1, but also compounds containing various amounts of water. Solvates of the compound represented by the above chemical formula 1 include both stoichiometric solvates and non-stoichiometric solvates.

[0030]

[0031] Meanwhile, examples of the fibrosis include interstitial lung disease (ILD), scleroderma, keloid, hypertrophic scar, non-alcoholic fatty liver disease, primary sclerosing cholangitis (PSC), primary biliary cholangitis (PBC), diabetic retinopathy, age-related macular degeneration (AMD), hypertrophic cardiomyopathy, myocardial infarction, muscular dystrophy, diabetic kidney disease, focal segmental glomerulosclerosis (FSGS), or inflammatory bowel disease. There is idiopathic pulmonary fibrosis (IPF), systemic sclerosis associated interstitial lung disease (SSc-ILD), or chronic fibrosing interstitial lung diseases with a progressive phenotype (PF-ILD).

[0032]

[0033] The term "prevention" of the present invention means any act of inhibiting or delaying the occurrence, spread, and recurrence of the disease by administering the composition of the present invention, and "treatment" means any act of improving or beneficially changing the symptoms of the disease by administering the composition of the present invention.

[0034]

[0035] The pharmaceutical composition of the present invention can be formulated into oral or parenteral dosage forms according to standard pharmaceutical practice. These dosage forms may contain, in addition to the active ingredient, pharmaceutically acceptable additives such as carriers, adjuvants, or diluents.

[0036]

[0037] Suitable carriers include, but are not limited to, saline, polyethylene glycol, ethanol, vegetable oils, and isopropyl myristate, and diluents include, but are not limited to, lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, and / or glycine. In addition, the compounds of the present invention can be dissolved in oils, propylene glycol, or other solvents commonly used in the preparation of injectable solutions. In addition, the compounds of the present invention can be formulated as ointments or creams for topical action.

[0038]

[0039] The pharmaceutical dosage forms of the compounds of the present invention can be used in the form of their pharmaceutically acceptable salts or solvates, and can be used alone or in combination with other pharmaceutically active compounds, as well as in an appropriate combination. For example, the pharmaceutical composition according to the present invention may additionally comprise another active ingredient used for the prevention or treatment of fibrosis. Examples of such other active ingredients include pirfenidone or nintedanib. In addition, when the pharmaceutical composition according to the present invention additionally comprises another active ingredient, the weight ratio of the compound represented by the above chemical formula 1, or a pharmaceutically acceptable salt thereof, to the other active ingredient is preferably 1:0.1 to 1:10.

[0040]

[0041] The compounds of the present invention can be formulated as injections by dissolving, suspending, or emulsifying the compounds in water-soluble solvents such as normal saline, 5% dextrose, or in water-insoluble solvents such as synthetic fatty acid glycerides, higher fatty acid esters, or propylene glycol. The formulations of the present invention can include conventional additives such as solubilizers, isotonic agents, suspending agents, emulsifiers, stabilizers, and preservatives.

[0042]

[0043] The pharmaceutical composition according to the present invention may be administered via oral or parenteral routes. Depending on the administration method, the composition according to the present invention may contain 0.001 to 99 wt%, preferably 0.01 to 60 wt%, of the compound represented by the above chemical formula 1, or a pharmaceutically acceptable salt thereof.

[0044]

[0045] The pharmaceutical composition of the present invention can be administered to mammals, including rats, mice, livestock, and humans, via various routes. Any route of administration is conceivable, including oral, rectal, intravenous, intramuscular, subcutaneous, intrauterine, or intracerebroventricular injection.

[0046] As described above, the pharmaceutical composition according to the present invention can be used in a specific therapeutic dosage and can be usefully used for the prevention or treatment of fibrosis.

[0047] Figure 1 shows the results of the lung function evaluation of Experimental Example 1-1.

[0048] Figure 2 shows the results of measuring the collagen content inside the lungs in Experimental Example 1-2.

[0049] Figure 3 shows the results of histopathological analysis of Experimental Example 1-3.

[0050] Figure 4 shows the results of the inflammatory cell infiltration analysis of Experimental Example 1-4.

[0051] Figure 5 shows the results of measuring body weight change in Experimental Example 1-5.

[0052] Figure 6 shows the change in blood concentration of the active ingredient of Experimental Example 3.

[0053] Figure 7 shows the change in plasma concentration for a single administration of Experimental Example 4.

[0054] Figure 8 shows the change in plasma concentration for multiple administrations of Experimental Example 4.

[0055] Figure 9 is a graph showing the correlation between AUC and the administered dose in Experimental Example 4.

[0056] Figure 10 shows the relationship between the time of occurrence of adverse effects such as nausea and vomiting and exposure in the body in Experimental Example 5.

[0057] Hereinafter, the present invention will be described in more detail with reference to the following examples. However, the following examples are intended only to illustrate the present invention and the scope of the present invention is not limited to these examples.

[0058]

[0059] Example

[0060] The following compound was prepared in the same manner as Example 40 of Korean Patent Registration No. 10-2084772, and was hereinafter named as 'active ingredient' or 'Example'.

[0061]

[0062] 1 H NMR (500 MHz, MeOD): δ 9.67 (s, 1H). 8.02 (d, 1H), 7.82 (d, 1H), 4.62 (m, 2H), 3.60 (m, 1H), 3.28 (m, 1H), 2.99 (m, 2H), 2.25 (m, 2H), 2.08 (m, 2H), 1.99 (m, 1H), 1.78 (m, 2H), 1.54 (m, 1H)

[0063]

[0064] Experimental Example 1: Animal study of anti-fibrotic efficacy by dose

[0065] In experimental animals acclimatized for more than 5 days, 70 to 100 uL of BLM solution (bleomycin 1 to 3 mg / kg) was administered into the lungs using a catheter. Depending on the purpose of the test, the test drug was administered 7 days after BLM administration, and the drug was administered orally for 2 weeks until the 21st day of BLM administration. The composition of the experimental group is as shown in Table 1 below, and the anti-fibrotic efficacy of the active ingredient was measured by measuring body weight, SpO2, hydroxyproline, and inflammatory cell count.

[0066]

[0067] Nintedanib (NIN) and pirfenidone (PID), which are standard of care (SoC) treatments for idiopathic pulmonary fibrosis, were used as control groups. The effective dose of NIN used was confirmed through in vivo efficacy data published by Boehringer Ingelheim, the developer of NIN, and the effective dose of PID was converted to mouse based on in vivo efficacy (rat) data and 2-week repeated toxic dose (rat) in FDA Pharmacology Review.

[0068] Group Drug No. of Animals (Male) Dose 1 Dose 2 Volume Administration method (mg / kg) (mg / kg) (SoC) (uL) G1 (NC) Saline 9N / AN / A 100 Oral G2 (PC) Saline 9N / AN / A 100 Oral G3 (Test) Active ingredient 93N / A 100 Oral G4 (Test) Active ingredient 910N / A 100 Oral G5 (Test) Active ingredient 930N / A 100 Oral G6 (Test) Nintedanib 9N / A 60 100 Oral G7 (Test) Pirfenidone 9N / A 200 100 Oral

[0069]

[0070] Experimental Example 1-1: Lung Function Evaluation

[0071] This lung function evaluation is the most direct and important evaluation index that has the greatest impact on the symptoms and quality of life of patients with pulmonary fibrosis. It is an experiment that can most directly show the effect of improving lung function in an animal model of pulmonary fibrosis by measuring the oxygen concentration in the body.

[0072]

[0073] On the 21st day, SpO2 was measured through the abdomen of the mouse using a device (Berry, Veterinary Pulse Oximeter), and the results are shown in Table 2 and Fig. 1.

[0074]

[0075] We attempted to confirm the oxygen permeability of the lungs by measuring the mouse abdominal SpO2, and confirmed that compared to the vehicle, the 'active ingredient' improved by more than 18% at 10 mg / kg and by more than 28% at 30 mg / kg, confirming that this is a similar improvement in oxygen permeability to the existing SoC material. The results confirm that there was a direct improvement in lung function when the 'active ingredient' was administered through the increase in oxygen permeability that was reduced during the lung fibrosis process.

[0076] NormalVehicleActive ingredientPirfenidoneNintedanib3 mg / kg10 mg / kg30 mg / kg200 mg / kg60 mg / kg99.8974.4475.0077.8981.3382.0078.67

[0077]

[0078] Experimental Example 1-2: Measurement of Total Collagen in Lung

[0079] Pulmonary fibrosis is a condition in which collagen accumulates in the lungs, causing them to harden. The main cause of pulmonary fibrosis is the accumulation of collagen, and the degree of fibrosis progression can be predicted by measuring the collagen content inside the lung tissue.

[0080]

[0081] In this experimental example, the INSOLUBLE Collagen Assay (Biocolor, S2000) was used for analysis. After sacrifice on day 21, frozen lung tissue was pulverized with 100 μL of Fragmentation Reagent, followed by the addition of 100 μL of 37% HCl and incubation at 65°C for 3 hours. The tube contents were shaken every 30 minutes to promote tissue disintegration. After centrifugation, the concentration was adjusted to 100 μL and collagen staining was performed to prepare the sample, and the absorbance was measured at 560 nm. The hydroxyproline value was measured as a ratio value compared to the normal group, and the results are shown in Table 3 and Figure 2.

[0082] NormalVehicleActive ingredientPirfenidoneNintedanib3 mg / kg10 mg / kg30 mg / kg200 mg / kg60 mg / kg96.11260.33253.56217.22171.33177.44239.78

[0083]

[0084] This experiment confirmed that the content of intrapulmonary collagen was significantly reduced at 10 and 30 mg / kg of the "active ingredient" compared to vehicle, a result similar to that of the existing SoC material, PID. This experiment demonstrated that the degree of pulmonary fibrosis was alleviated when the active ingredient was administered.

[0085]

[0086] Experimental Example 1-3: Histopathological (Ashcroft Score) Analysis

[0087] The degree of fibrosis and inflammation in lung tissue was visually observed under a microscope, and the degree of fibrosis in lung tissue was measured using the Fibrotic Index (FI), based on normalized criteria. A higher FI value indicates a more severe degree of fibrosis and disease, while a lower value indicates a more alleviated condition.

[0088]

[0089] On the 21st day, lung tissue was isolated and stained using H&E and MT stain, observed at 200X magnification, and the observed results were scored using Ashcroft scoring (Hubner et al., 2008). The fibrosis index was calculated as the sum of the modified Ashcroft field scores divided by the number of examined fields, and is shown in Table 4 and Figure 3.

[0090] Group Number of individuals Fibrotic Index (±SEM) Normal 90.1 (±0.1) Vehicle 95.2 (±0.2) Active ingredient 3 mg / kg 94.6 (±0.3) Active ingredient 10 mg / kg 92.7 (±0.6) Active ingredient 30 mg / kg 92.8 (±0.4) Pirfenidone 200 mg / kg 94.1 (±0.3) Nintedanib 60 mg / kg 93.1 (±0.6)

[0091]

[0092] For 10 and 30 mg / kg of the ‘active ingredient’, a tissue improvement effect equivalent to or greater than NIN was confirmed, and a tissue improvement effect significantly superior to PID was confirmed.

[0093]

[0094] Experimental Example 1-4: Inflammation cell count analysis

[0095] Since fibrosis is a chronic inflammatory disease with excessive collagen deposition, the infiltration of inflammatory cells was analyzed to determine the degree of inflammation in lung tissue.

[0096]

[0097] On the 21st day of administration, BALF (Bronchoalveolar lavage fluid) cells obtained through airway lavage of the mouse at sacrifice were diluted with 1.05 X PBS and attached to a slide. The slide was then dipped and removed in the order of 1, 2, and 3 diff quick stain solutions for 30 seconds each and stained. Counting was performed based on 500 cells. Macrophages are the largest in size and have mononuclei that stain blue. Neutrophils and eosinophils form multinucleated cells, but eosinophils are distinguished from neutrophils by their red eosin staining. Lymphocytes are small in size and mononucleated with very little cytoplasm. The total cells were counted and converted to a percentage, which is shown in Table 5 and Figure 4.

[0098] NormalVehicleActive ingredientPirfenidoneNintedanib3 mg / kg10 mg / kg30 mg / kg200 mg / kg60 mg / kgMacrophage0.426.967.805.604.915.015.61Neutrophil0.002.121.741.540.820.681.29Lymphocyte0.000.870.790.510.460.460.68Total cell0.429.9310.327.666.196.147.57

[0099]

[0100] Through this experiment, it was confirmed that the total cell count was improved by more than 20% at 10 mg / kg of the 'active ingredient' and more than 30% at 30 mg / kg compared to the vehicle, indicating an improvement in lung tissue inflammation. In particular, neutrophil cells are inflammatory cells that show a high proportion of lung fibrotic inflammation, and the number of these cells was reduced by more than 20% at 10 mg / kg of the 'active ingredient' and more than 60% at 30 mg / kg compared to the vehicle, confirming a significant improvement in the proportion of major inflammatory cells. This is equivalent to the existing SoC material, PID, and can be seen to be a superior effect than NIN.

[0101]

[0102] Experimental Example 1-5: Measuring Changes in Body Weight

[0103] Body weight is an indirect indicator of the degree of improvement in the overall body condition of an animal model. If the degree of weight loss is small, it can be assumed that the overall body condition or symptoms of the disease have improved.

[0104] Body weight was measured on the 21st day after completion of medication using a scale, and the results are shown in Table 6 and Figure 5 below.

[0105] NormalVehicleActive ingredientPirfenidoneNintedanib3 mg / kg10 mg / kg30 mg / kg200 mg / kg60 mg / kg13.22-23.33-22.44-19.11-16.78-14.67-19.44

[0106]

[0107] It was confirmed that the body weight loss was improved by 10 mg / kg and 30 mg / kg compared to the vehicle group, which confirmed that the improvement effect was similar to or greater than that of the SoC substance, and it was confirmed that the overall symptoms improved when the 'active ingredient' was administered.

[0108]

[0109] Through the above series of experimental results, it can be confirmed that when the 'active ingredient' is administered to mice once a day at 10 mpk, the therapeutic effect is equivalent to that of the existing standard treatment for IPF.

[0110]

[0111] Experimental Example 2: Two-week repeated dose toxicity test in mice

[0112] ICR mice acclimatized for more than 7 days were orally administered the active ingredient for 2 weeks. The composition of the experimental groups is shown in Table 7 below, and body weight, general symptom observations, hematology / blood chemistry tests, organ weight measurements, and histopathological examinations were performed.

[0113] Group Sex Number of animals (animals) Dosage (mg / kg / day) Dosage (mL / kg / day) G1M6 Sterile distilled water -10 G2M6 Active ingredient 30 10 G3M6 Active ingredient 60 10 G4M6 Active ingredient 120 → 90 1) 101) If death is confirmed in the high-dose group during the initial administration (within a few days of administration) or if the animal's condition appears unlikely to last 2 weeks, the dose is reduced and administered for the remaining period.

[0114]

[0115] After two weeks of administration, all animals were necropsied, and the weights of each organ were measured. No toxicity or significant abnormalities were observed in any of the results measured after two weeks of repeated administration. However, as shown in Table 8 below, a slight weight decrease was observed only in the spleen and thymus among all organs in the group administered at doses of 30 mg / kg / day or higher.

[0116] Dose (mg / kg)03060120→90SPLEEN0.080±0.0180.059±0.0110.070±0.0150.046±0.021% to BODY WEIGHT0.28±0.070.22±0.040.25±0.050.17±0.08THYMUS0.034±0.0080.026±0.0080.029±0.0060.015±0.012% to BODY WEIGHT0.12±0.020.10±0.030.11±0.020.05±0.04

[0117]

[0118] Although these results were judged to be unrelated to toxicity, the effective dose of the active ingredient was set to 10 mg / kg to prevent potential toxicity risks during long-term administration based on the weight loss tendency of the spleen and thoracic organs.

[0119]

[0120] Experimental Example 3: Animal Pharmacokinetic / Pharmacodynamic Analysis

[0121] After a single oral administration of the active ingredient in ICR mice, a kinetic study was conducted as shown in Table 9. The blood drug concentration over time obtained through LC-MS / MS was used to calculate pharmacokinetic parameters using Excel® and WinNonlin6.1 software, and the changes in the blood concentration of the active ingredient are shown in Table 10 and Figure 6.

[0122] Experimental animal ICR mice (male) Test substance Active ingredient Administration route Oral Number of administrations Single dose (mg / kg) 10 Amount of administered liquid (mL / kg) 10 Blood collection time (hr) 0.08 3, 0.25, 0.5, 1, 2, 4, 6, 8, 24 Excipient saline Number of test animals 6 (n=3, cross-blood collection)

[0123]

[0124] Test substance active ingredient administration dose (mg / kg) 10t 1 / 2 (hr)2.50 ± 2.48T max (hr)2.67 ± 1.15C max(ug / mL)0.263 ± 0.032AUC 0-t (hr·ug / mL)1.07 ± 0.21AUCinh (hr·ng / mL)1190

[0125]

[0126] Manufacturing Example 1: Manufacturing of enteric-coated capsules containing active ingredients

[0127] The active ingredient in hydrochloride form was filled into capsules according to dosage using Vcaps® enteric-coated capsules, containing only the active ingredient without any additional excipients.

[0128]

[0129] Manufacturing Example 2: Manufacturing of enteric-coated tablets containing active ingredients

[0130] The active ingredient hydrochloride form was mixed with microcrystalline cellulose, lactose monohydrate, crospovidone, and magnesium stearate, and the mixture was compressed into a plate shape using a dry granulator, and then ground with an oscillator to produce dry granules. Microcrystalline cellulose, lactose monohydrate, and magnesium stearate were further mixed into the granules, and compression molded to produce tablets, which were then subjected to enteric coating to complete the process.

[0131]

[0132] Experimental Example 4: Human Pharmacokinetic / Pharmacodynamic Analysis

[0133] To confirm the safety / tolerance / pK in humans, a randomized, double-blind, placebo-controlled clinical trial was designed with 72 healthy adults as shown in Table 11, using a single-dose, stepwise increase in dose. The formulation used in the clinical trial was prepared using the same method as in Manufacturing Examples 1 and 2 above.

[0134] Classification GroupActive ingredientDosage formAdministration methodSingle doseSAD1100 mgEnteric-coated capsuleSingle oral administration on an empty stomachSAD2300 mgSAD3600 mgSAD4500 mgEnteric-coated tabletMultiple doseMAD125 mgOral administration twice a day on an empty stomach for 13 consecutive days, then single administration on the morning of the 14th dayMAD250 mgMAD3100 mgMAD4200 mg

[0135]

[0136] The above clinical results did not reveal any significant abnormalities or side effects.

[0137]

[0138] As a result of the pK confirmation for a single administration, as can be confirmed in Table 12 and Figure 7 below, the maximum plasma concentration was recorded within 1.5 to 8 hours after administration, the geometric mean half-life was in the range of 6.91 to 9.90 hours, and it was confirmed that the plasma concentration increased in a dose-proportional relationship.

[0139] Item (GCV%)Active ingredient 100 mg (N=6)Active ingredient 300 mg (N=6)Active ingredient 600 mg (N=6)Active ingredient 500 mg (N=6)C max (ng / mL)123.96 (37.72)276.80 (66.21)213.43 (433.75)513.56 (41.27)t max (h)*3.00 (1.50, 4.00)2.50 (1.50, 8.00)2.00 (1.50, 5.00)3.01 (0.50, 5.00)AUC 0-last (h*ng / mL)594.10 (61.23)1478.84 (48.07)1008.65 (625.00)2282.37 (54.92)AUC 0-∞ (h*ng / mL)639.07 (58.10)1525.99 (47.06)1922.64 (222.08)2339.20 (53.27)T 1 / 2(h)6.91 (39.98)8.33 (24.57)9.90 (71.50)7.80 (13.49)Vz / F (L)1560.44 (34.98)2361.87 (41.96)4459.07 (84.14)2406.62 (48.04)CL / F (L / h)156.48 (58.10)196.59 (47.06)312.07 (222.08)213.75 (53.27)MRT (h)9.26 (33.69)9.75 (21.76)9.79 (25.60)9.57 (27.28)GCV% = Geometric coefficient of variationNotes: t max represented as median; min, max.

[0140]

[0141] In addition, as confirmed in Table 13 and Figure 8 as a result of the 14th day pK confirmation for multiple administrations, the maximum plasma concentration was recorded within 1.5 to 5 hours at steady state after administration, the geometric mean half-life was in the range of 4.4 to 12.35 hours, and the geometric mean dosing interval period in vivo exposure (AUCτ) was in the range of 183.32-3012.35 h·ng / mL. It was confirmed that the plasma concentration had a linear relationship with the dose within the dose range of 25 mg to 200 mg of the active ingredient.

[0142] Item (GCV%) Active ingredient 25 mg T (N=6) Active ingredient 50 mg T (N=6) Active ingredient 100 mg T (N=6) Active ingredient 200 mg T (N=6)Day 14C max, ss (ng / mL)36.50 (66.19)102.61 (67.99)151.27 (65.36)595.35 (36.32)t max, ss(h)*2.00 (1.50, 4.00)3.00 (3.00, 5.00)3.00 (1.50, 4.00)2.00 (1.50, 5.00)AUCτ (h*ng / mL)183.32 (76.35)429.09 (128.63)617.40 (60.53)3012.35 (48.42)AUC 0-∞ (h*ng / mL)300.73 (102.79)565.34 (232.53)887.97 (60.55)4747.29 (64.90)T 1 / 2 (h)7.63 (46.84)4.40 (101.38)9.26 (22.69)12.35 (34.54)Vz / F ss (L)1433.79 (61.75)738.97 (34.90)2163.04 (63.58)1182.53 (48.47)CL / F ss (L / h)122.65 (79.49)116.53 (128.63)161.97 (60.53)66.39 (48.42)MRT (h)10.53 (31.65)8.44 (68.43)10.17 (9.35)12.07 (26.90)Accumulation ratio based on AUCτ1.94 (26.34)2.33 (31.42)1.88 (35.07)3.75 (36.01)Abbreviations: GCV% = Geometric coefficient of variation; n = Number of subjects; SD = Standard Deviation; T = Tablet.Notes: * t max, ss represented as median; min, max. MRT, AUCτ, CL / F ss , and Vz / F ss are considering a 12 hour dosing interval.

[0143]

[0144] Accordingly, the optimal pharmacokinetic model parameters that can explain the concentration changes at all doses observed in the blood drug concentrations obtained in the above human multiple-dose clinical trial were estimated using Excel® and WinNonlin8.1 software, and the exposure levels in the body in humans when various doses are administered orally were predicted. The main results are as shown in Table 14 below.

[0145] Dose (mg)Parameter50100150200T 1 / 2 (h)9.7531959.7531959.7531959.753195T max (h)2.8032.8032.8032.803C max (ng / mL)38.27176.5419114.813153.084CL / F (mL / hr)215856.7215856.7215856.7215856.5Vd / F (mL)3037295303729530372953037293AUC tau (hr*ng / mL)231.6352463.2704694.9055926.5413AUC tau _24hr (hr*ng / mL)463.2704926.54071389.8111853.08256

[0146]

[0147] From the rat experimental model experiment for pulmonary fibrosis of Experimental Example 2 above, 10 mg / kg administered once a day is the effective dose for rats, and from Experimental Example 3, rat plasma AUC at the effective dose inf The value was confirmed to be 1190 hr·ng / mL. Therefore, the AUC is the same level as the effective dose in rats in humans. inf In order to predict the dose expected to show the value, the correlation with AUC according to the administered dose was examined as in Figure 9, and as a result, it was confirmed that 150 mg administration is necessary when administering twice a day to humans.

[0148]

[0149] Experimental Example 5: Human Administration Safety / Drug Tolerance Analysis

[0150] The safety and drug resistance of the compound of the present invention were confirmed when administered to humans using the same method as in Experimental Example 4 above.

[0151]

[0152] As a result of checking the safety and tolerability of a single dose, the most frequently occurring adverse reactions were gastrointestinal adverse reactions including nausea, vomiting, diarrhea, and abdominal pain. Among these, nausea and vomiting were evaluated to have a greater impact on safety and drug tolerability, and it was confirmed that this was caused by the active ingredient itself. As a result of checking the relationship between the time of occurrence of these nausea and vomiting side effects and exposure in the body, as shown in Figure 10, it was confirmed that most of them occurred before the blood concentration in the body increased. This means that the nausea and vomiting that occur when the active ingredient is administered is not a vomiting center activation pathway through stimulation of the chemoreceptor-induced site by blood exposure in the body, but rather an activation of the vomiting center by acting on the gastrointestinal tract to stimulate the vagus nerve.

[0153]

[0154] Therefore, it can be seen that the dosage of dividing the effective dose into two times a day to reduce the proportion of vagus nerve stimulation acting on the gastrointestinal tract is more appropriate for this active ingredient.

[0155]

[0156] Experimental Example 6: Confirming the therapeutic effect in humans

[0157] To determine the safety and efficacy of the active ingredient in patients with idiopathic pulmonary fibrosis, a randomized, double-blind, placebo-controlled clinical trial was designed, targeting either the standard treatment group or the non-administered group, as shown in Table 15. To determine the safety and tolerability of the active ingredient, the patients were administered the active ingredient for 24 weeks and compared with the placebo. To determine the therapeutic effect of the active ingredient on idiopathic pulmonary fibrosis, the primary efficacy endpoint was the rate of decline in forced vital capacity (FVC) from baseline over 24 weeks after administering the active ingredient. Secondary efficacy endpoints included 1) time to IPF disease progression, including respiratory-related mortality or hospitalization, acute exacerbation of IPF, a relative decline of ≥10% of the predicted value in FVC, and an absolute decline of ≥15% of the predicted value in diffusing capacity for carbon monoxide (DLCO) adjusted for hemoglobin; 2) time to first unplanned hospitalization for any cause during 24 weeks; 3) Baseline change in functional exercise capacity assessed by 6-minute walk test (6MWT) distance at 24 weeks; 4) Baseline change in DLCO (adjusted for hemoglobin) at 24 weeks; 5) Categorical assessment of absolute change from baseline in percent predicted value of FVC at 24 weeks; 6) Baseline change in quantitative chest high-resolution CT (HRCT) at 24 weeks; 7) Baseline change in patient-reported outcomes (PROs) measured by the St George's Respiratory Questionnaire, SGRQ, and Living with Idiopathic Pulmonary Fibrosis (L-IPF) at 24 weeks. Exploratory efficacy endpoints include 1) Baseline change in IPF-specific biomarkers at 24 weeks; and 2) Baseline change in blood biomarkers at 24 weeks.As safety evaluation variables, 1) incidence of adverse reactions after administration of the active ingredient; 2) physical examination; 3) 12-lead electrocardiogram test; 4) vital signs; 5) clinical laboratory performance tests are evaluated.

[0158]

[0159] The target group is divided into patients receiving pirfenidone as a standard treatment, patients receiving nintedanib, and patients not receiving any other treatment. Each patient group will be administered the active ingredient or a placebo to determine the efficacy. If adverse reactions due to the active ingredient occur, the patient's response will be closely monitored and a dose reduction will be considered.

[0160] Previously administered treatmentActive ingredient 150 mg, BIDActive ingredient 100 mg, BIDPlacebo (0 mg), BIDPirfenidone (mg, administered three times a day)801801801534534534267267267600600600400400400200200Nintedanib (mg, administered twice a day)150150150100100100No treatment administered (mg)000

Claims

1. 1 A pharmaceutical composition for the prevention or treatment of fibrosis, comprising a compound represented by the following chemical formula 1, or a pharmaceutically acceptable salt thereof, for administration of 100 to 150 mg twice daily (BID): [Chemical Formula 1] .

2. In paragraph 1, The above fibrosis may be caused by interstitial lung disease (ILD), scleroderma, keloid, hypertrophic scar, non-alcoholic fatty liver disease, primary sclerosing cholangitis (PSC), primary biliary cholangitis (PBC), diabetic retinopathy, age-related macular degeneration (AMD), hypertrophic cardiomyopathy, myocardial infarction, muscular dystrophy, diabetic kidney disease, focal segmental glomerulosclerosis (FSGS), or inflammatory bowel disease, IBD) Pharmaceutical composition.

3. In paragraph 2, The above interstitial lung disease is idiopathic pulmonary fibrosis (IPF), systemic sclerosis associated interstitial lung disease (SSc-ILD), or chronic fibrosing interstitial lung diseases with a progressive phenotype (PF-ILD). Pharmaceutical composition.

4. In paragraph 1, The pharmaceutically acceptable salt is hydrochloride, Pharmaceutical composition.

5. In paragraph 1, The pharmaceutical composition further comprises another effective ingredient used for the prevention or treatment of fibrosis. Pharmaceutical composition.

6. In paragraph 5, Other active ingredients used for the prevention or treatment of the above fibrosis are pirfenidone or nintedanib, Pharmaceutical composition.

Images