WO2023073600A1

WO2023073600A1 - Compositions for preventing or treating idiopathic pulmonary fibrosis (ipf)

Info

Publication number: WO2023073600A1
Application number: PCT/IB2022/060326
Authority: WO
Inventors: Nina Ha; Min Cheol Kim; Dae Kwon Bae; Suk Jin Lee; Dong-hyeon SUH; Ji Yeon Baek
Original assignee: Chong Kun Dang Corp
Current assignee: Chong Kun Dang Corp
Priority date: 2021-10-29
Filing date: 2022-10-27
Publication date: 2023-05-04
Anticipated expiration: 2024-04-29
Also published as: KR20230062785A; TW202328118A

Abstract

The present disclosure relates to a pharmaceutical composition for preventing or treating idiopathic pulmonary fibrosis, comprising a compound represented by formula I, optical isomers thereof or pharmaceutically acceptable salts thereof as an effective ingredient, a method for preventing or treating idiopathic pulmonary fibrosis using the compound, a use of the compound for preventing or treating idiopathic pulmonary fibrosis, and a use of the compound in preparing a medicament for preventing or treating idiopathic pulmonary fibrosis.

Description

Compositions for preventing or treating idiopathic pulmonary fibrosis (IPF) Technical Field

The present disclosure relates to a pharmaceutical composition for preventing or treating idiopathic pulmonary fibrosis, comprising a compound represented by formula I, optical isomers thereof or pharmaceutically acceptable salts thereof as an effective ingredient, a method for preventing or treating idiopathic pulmonary fibrosis using the compound, a use of the compound for preventing or treating idiopathic pulmonary fibrosis, and a use of the compound in preparing a medicament for preventing or treating idiopathic pulmonary fibrosis. Background

Idiopathic pulmonary fibrosis (IPF) is a disease in which the lung parenchyma becomes fibrous due to an abnormal tissue repair mechanism after damage to alveolar epithelial cells for unknown reasons. The IPF shows symptoms such as chronic dry cough, shortness of breath, etc., and is a disease with a poor prognosis, which usually has a survival period of about 3-5 years only after being diagnosed due to the occurrence of the symptoms. It is known that an incidence rate is higher in men over the age of 50.

Currently, two drugs, nintedanib and pirfenidone, are mainly used for the treatment of idiopathic pulmonary fibrosis, but the drugs slow only a progression of the disease without improving a survival rate. Accordingly, there is an urgent need for developing a drug capable of effectively treating idiopathic pulmonary fibrosis.

[Related Art Reference]

[Patent Documents]

(Patent Document 1) Korean Unexamined Patent Application Publication No.10-2014-0128886

Detailed Description of the Invention

Technical Problem

The present disclosure provides a pharmaceutical composition for preventing or treating idiopathic pulmonary fibrosis, including a compound represented by formula I, optical isomers thereof or pharmaceutically acceptable salts thereof as an effective ingredient.

The present disclosure provides a method for preventing or treating idiopathic pulmonary fibrosis, including administering a compound represented by formula I, optical isomers thereof or pharmaceutically acceptable salts thereof into an individual. The present disclosure provides a use of a compound represented by formula I, optical isomers thereof or pharmaceutically acceptable salts thereof for preventing or treating idiopathic pulmonary fibrosis.

The present disclosure provides a use of a compound represented by formula I, optical isomers thereof or pharmaceutically acceptable salts thereof in preparing a medicament for preventing or treating idiopathic pulmonary fibrosis.

Technical Solution

This is described in detail as follows. Meanwhile, each description and embodiment disclosed in the present disclosure may be also applied to other descriptions and embodiments thereof, respectively. In other words, all the combinations of various elements disclosed in the present disclosure fall within the scope of the present disclosure. Also, it cannot be seen that the scope of the present disclosure is limited to the specific description described below.

The present disclosure provides a pharmaceutical composition for preventing or treating idiopathic pulmonary fibrosis, including a compound represented by a following formula I, optical isomers thereof or pharmaceutically acceptable salts thereof as an effective ingredient.

[Formula I]

Xa and Xb are each independently CH or N,

Li and L2 are each independently hydrogen, halogen, -CF3, or -C1-3 straight or branched chain alkyl,

Q is C(=O), S(=O)₂, S(=O) or C(=NH),

Y is selected from a following group:

M is C, N, O, S or S(=O)2 (at this time, if M is C, 1 and m are 1; if M is N, 1 is 1 and m is 0; and if M is O, S or S(=O)2, 1 and m are 0),

Rai and Ra2 are each independently hydrogen; hydroxy; -Ci-4 straight or branched chain alkyl, which is unsubstituted or substituted with at least one halogen; -Ci-4 straight or branched chain alcohol; benzhydryl; -Ci-4 straight or branched chain alkyl, which is substituted with a saturated or unsaturated five to seven-membered heterocyclic compound having one to three heteroatoms out of N, O or S as a ring member (at this time, the heterocyclic compound may be unsubstituted or at least one hydrogen may be optionally substituted with OH, OCH3, CH3, CH2CH3 or halogen); a saturated or unsaturated five to seven-membered heterocyclic compound having one to three heteroatoms out of N, O or S as a ring member (at this time, the heterocyclic compound may be unsubstituted or at least one hydrogen may be optionally substituted with OH, OCH3, CH3, CH2CH3 or halogen); phenyl, which is unsubstituted or in which at least one hydrogen is substituted with halogen, Ci-4 alkoxy, C1-2 alkyl or hydroxy; benzyl, which is unsubstituted or in which at least one hydrogen is substituted with halogen, Ci-4 alkoxy, C1-2 alkyl or hydroxy; -S(=O)2CH3; halogen; -C1-6 straight or branched chain alkoxy; -C2-6 alkoxyalkyl; -C(=O)R_x, in which R_x is

C1-3 straight or branched chain alkyl or C3-10 cycloalkyl;

, in which Rc and Rd are independently hydrogen or C1-3 straight or branched chain alkyl;

n is an integer of 0, 1 or 2,

Rb is hydrogen; hydroxy; -C1-6 straight or branched chain alkyl, which is unsubstituted or in which at least one hydrogen is substituted with halogen; -C(=O)CH3; -Ci-4 straight or branched chain hydroxyalkyl; -C1-6 straight or branched chain alkoxy; -C2-6 straight or branched chain alkoxyalkyl; -CF3; halogen;

Re and Rf are each independently hydrogen or -C1-3 straight or branched chain alkyl, and

Z is selected from a following group:

Pa and Pb are each independently

hydrogen; hydroxy; -Ci-4 straight or branched chain alkyl, which is unsubstituted or in which at least one hydrogen is substituted with halogen; halogen; -CF3; -OCF3; -CN; -C1-6 straight or branched chain alkoxy; -C2-6 straight or branched chain alkyl alkoxy; -CH2F; or -C1-3 alcohol, in which C⁸ is a ring selected from phenyl, pyridine, pyrimidine, thiazole, indole, indazole, piperazine, quinoline, furan, tetrahydropyridine, piperidine or a following group:

x, y and z are each independently an integer of 0 or 1, and

Rgi, Rg2 and R_g3 are each independently selected from hydrogen; hydroxy; -C1-3 alkyl; -CF3; -C1-6 straight or branched chain alkoxy; -C2-6 straight or branched chain alkyl alkoxy; -C(=O)CH3; -Ci-4 straight or branched chain hydroxyalkyl; -N(CH3)2; halogen; phenyl; -S((=O)₂)CH3; or a following group:

In the present disclosure, the compound represented by the above formula I is a compound represented by a following formula la:

[Formula la]

wherein

Y is selected from a following group:

in which M, 1, m, n, Rai, Ra2, and Rb are each the same as defined in the above formula I,

Pa and Pb are each independently hydrogen; hydroxy; -Ci-4 straight or branched chain alkyl, which is unsubstituted or in which at least one hydrogen is substituted with halogen; halogen; -CF3; -OCF3; -CN; -C1-6 straight or branched chain alkoxy; -C2-6 straight or branched chain alkyl alkoxy; -CH2F; or-Ci-3 alcohol.

According to one embodiment aspect of the present disclosure, Y is selected from a following group:

in which n and Rb are each the same as defined in the above formula I,

Z / i •s Ph ^b , and P_a and Pb are each independently hydrogen; halogen; -CF3; or -C1-6 straight or branched chain alkoxy.

In the present disclosure, “halogen” is F, Cl, Br or I.

According to a specific embodiment of the present disclosure, the compound represented by the above formula I may be a compound described in a following table.

In the present disclosure, the compound represented by above formula I may be prepared by a method disclosed in Korean Unexamined Patent Application Publication No. 10-2014-0128886, but is not limited thereto.

In the present disclosure, the compound represented by above formula I may contain at least one asymmetric carbon, and thus may be present as an enantiomer mixture including a racemic mixture, a single enantiomer (optical isomer), a mixture of diastereomers, and a single diastereomer. Such isomer may be separated by split according to the related art, for example, column chromatography, HPLC or the like. Alternatively, the isomer may be stereospecifically synthesized by using a known array of optically pure starting materials and/or reagents. Particularly, said isomer may be an optical isomer (enantiomer).

In the present disclosure, the term “pharmaceutically acceptable” may refer to the one which is physiologically acceptable and does not conventionally cause gastrointestinal disturbance, an allergic response such as dizziness or other responses similar thereto, when being administered to an individual.

The pharmaceutically acceptable salts according to the embodiments of the present disclosure may be prepared by a conventional method known to those skilled in the art.

The pharmaceutically acceptable salts according to the embodiment of the present disclosure may include, for example, inorganic ion salts prepared from calcium, potassium, sodium, magnesium, etc.; inorganic acid salts prepared from hydrochloric acid, nitric acid, phosphoric acid, bromic acid, iodic acid, perchloric acid, sulfuric acid, hydroiodic acid, etc.; organic acid salts prepared from acetic acid, trifluoroacetic acid, citric acid, maleic acid, succinic acid, oxalic acid, benzoic acid, tartaric acid, fumaric acid, mandelic acid, propionic acid, lactic acid, glycolic acid, gluconic acid, galacturonic acid, glutamic acid, glutaric acid, glucuronic acid, aspartic acid, ascorbic acid, carbonic acid, vanillic acid, etc.; sulfonic acid salts prepared from methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p- toluenesulfonic acid, naphthalene sulfonic acid, etc.; amino acid salts prepared from glycine, arginine, lysine, etc.; amine salts prepared from trimethylamine, tri ethylamine, ammonia, pyridine, picoline, etc.; and the like, but are not limited thereto. In the embodiments of the present disclosure, salts may include hydrochloric acid, trifluoroacetic acid, citric acid, bromic acid, maleic acid, phosphoric acid, sulfuric acid or tartaric acid.

In the present disclosure, the term "idiopathic pulmonary fibrosis (IPF)" may refer to a disease in which the lung parenchyma becomes fibrous due to an abnormal tissue repair mechanism after damage to alveolar epithelial cells for unknown reasons.

In the present disclosure, the term “prevention” may refer to all the acts, which inhibit or delay the occurrence of a disease by administering the compound of formula I of the present disclosure, optical isomers thereof or pharmaceutically acceptable salts thereof.

In the present disclosure, the term “treatment” may refer to all the acts, by which a symptom of an individual likely to develop or suffering from a disease gets better or takes a favorable turn by administering the compound of formula I of the present disclosure, optical isomers thereof or pharmaceutically acceptable salts thereof.

The compound represented by formula I of the present disclosure, optical isomers thereof or pharmaceutically acceptable salts thereof may be advantageously used in preventing or treating idiopathic pulmonary fibrosis.

A pharmaceutical composition including the compound represented by formula I of the present disclosure, optical isomers thereof, or pharmaceutically acceptable salts thereof as an active ingredient may be advantageously used in preventing or treating idiopathic pulmonary fibrosis.

In this regard, in one specific embodiment of the present disclosure, it was confirmed that the compound represented by formula I of the present disclosure, optical isomers thereof or pharmaceutically acceptable salts thereof inhibit the expression of TGF- pi-induced fibrotic proteins, proCPLl Al and LOXL2 (FIG. 1).

In addition, it was confirmed that in mice with pulmonary fibrosis induced by BLM, the compound represented by formula I of the present disclosure, optical isomers thereof or pharmaceutically acceptable salts thereof recover a reduced body weight (FIG. 2); reduce the number of increased total cells, neutrophils, lymphocytes and macrophages, so as to suppress the infiltration of inflammatory cells (FIG. 3); decrease an increased lung weight (FIG. 4); suppress the expression of increased fibrosis genes, COK1A1, FN1, PAI1, TIMP-1 and SPP1 (FIG. 5); and reduce an increased Ashcroft score and de novo collagen 1A1 deposition, so as to exhibit an anti-fibrotic effect (FIGS. 6 and 7).

Furthermore, it was confirmed that the compound represented by formula I of the present disclosure, optical isomers thereof or pharmaceutically acceptable salts thereof inhibit collagen contraction induced by TGF-pi (FIGS. 8 and 9); inhibit the migration of fibroblasts, DHLF-IPF cells (FIGS. 10 and 11); and inhibit the proliferation of cells induced by PDGF- BB (FIG. 12), so as to exhibit an anti-fibrotic effect.

The compound represented by formula I of the present disclosure, optical isomers thereof or pharmaceutically acceptable salts thereof may show an effect of preventing or treating idiopathic pulmonary fibrosis at a level that is similar to or substantially the same as or superior to a conventionally known drug for preventing or treating idiopathic pulmonary fibrosis.

A pharmaceutical composition including the compound represented by formula I of the present disclosure, optical isomers thereof, or pharmaceutically acceptable salts thereof as an active ingredient may show an effect of preventing or treating idiopathic pulmonary fibrosis at a level that is similar to or substantially the same as or superior to a conventionally known drug for preventing or treating idiopathic pulmonary fibrosis.

The pharmaceutical composition according to the embodiments of the present disclosure may further include at least one pharmaceutically acceptable carrier, in addition to the compound represented by the above formula I, optical isomers thereof or pharmaceutically acceptable salts thereof. The pharmaceutically acceptable carrier may be the one which is conventionally used in the art, specifically including, but not limited thereto, lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinyl pyrrolidine, cellulose, water, syrup, methylcellulose, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, or mineral oil. The pharmaceutical composition according to the embodiments of the present disclosure may further include lubricants, humectants, sweetening agents, flavoring agents, emulsifiers, suspending agents, preservatives, dispersing agents, stabilizing agents, etc., in addition to the above ingredients. In addition, the pharmaceutical composition according to the embodiments of the present disclosure may be formulated into an oral dosage form such as a tablet, powder, granule, pill, capsule, suspension, emulsion, liquid for internal use, oiling agent, syrup, etc., as well as a form of external application, suppository or sterile solution for injection, by using pharmaceutically acceptable carriers and excipients and thus may be prepared in a unit dose form or prepared by being inserted into a multi-dose container. Such preparations may be prepared according to a conventional method used for formulation in the art or a method disclosed in Remington's Pharmaceutical Science (19^th ed., 1995), and may be formulated into various preparations depending on each disease or ingredient.

A non-limiting example of preparations for oral administration using the pharmaceutical composition of the present disclosure may include tablets, troches, lozenges, water-soluble suspensions, oil suspensions, prepared powders, granules, emulsions, hard capsules, soft capsules, syrups, elixirs or the like. To formulate the pharmaceutical composition according to the embodiments of the present disclosure into preparation for oral administration, the followings may be used: binders such as lactose, saccharose, sorbitol, mannitol, starch, amylopectin, cellulose, gelatin or the like; excipients such as dicalcium phosphate, etc.; disintegrants such as maize starch, sweet potato starch or the like; lubricants such as magnesium stearate, calcium stearate, sodium stearyl fumarate, polyethylene glycol wax, or the like; etc., in which sweetening agents, flavoring agents, syrups, etc. may also be used. Furthermore, in the case of the capsules, liquid carriers such as fatty oil, etc. may be further used in addition to the above-mentioned materials.

A non-limiting example of parenteral preparations using the pharmaceutical composition according to the embodiments of the present disclosure may include injectable solutions, suppositories, powders for respiratory inhalation, aerosols for spray, ointments, powders for application, oils, creams, etc. To formulate the pharmaceutical composition according to the embodiments of the present disclosure into preparation for parenteral administration, the following may be used: sterilized aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried preparations, external preparations, etc. As said nonaqueous solvents and suspensions, the following may be used, but without limitation thereto: propylene glycol, polyethylene glycol, vegetable oils such as olive oil, injectable esters such as ethyl oleate, etc.

The pharmaceutical composition according to the embodiments of the present disclosure may be subjected to oral administration or parenteral administration according to a targeted method, preferably oral administration, but is not limited thereto.

A daily dosage of the compound represented by formula I of the present discosure, optical isomers thereof or pharmaceutically acceptable salts thereof may be particularly about 0.1 to about 10,000 mg/kg, about 1 to about 8,000 mg/kg, about 5 to about 6,000 mg/kg, or about 10 to about 4,000 mg/kg, and more particularly about 50 to about 2,000 mg/kg, but is not limited thereto and may be also administered once a day or several times a day by dividing the daily dosage of the compound.

A pharmaceutically effective dose and an effective dosage of the pharmaceutical composition according to the embodiments of the present disclosure may vary depending on a method for formulating the pharmaceutical composition, an administration mode, an administration time, an administration route, and/or the like, and may be diversified according to various factors including a type and degree of reaction to be achieved by administration of the pharmaceutical composition, a type of an individual for administration, the individual’s age, weight, general health condition, disease symptom or severity, gender, diet and excretion, ingredients of other drug compositions to be used for the corresponding individual at the same time or different times, etc., as well as other similar factors well known in a pharmaceutical field, and those skilled in the art may easily determine and prescribe an effective dosage for the intended treatment.

The pharmaceutical composition according to the embodiments of the present disclosure may be administered once a day or several times a day. The pharmaceutical composition according to the embodiments of the present disclosure may be administered as an individual therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with a conventional therapeutic agent. Considering all the above factors, the pharmaceutical composition according to the embodiments of the present disclosure may be administered in such an amount that a maximum effect may be achieved by a minimum amount without a side effect, and such amount may be easily determined by those skilled in the art to which the present disclosure pertains.

The pharmaceutical composition according to the embodiments of the present disclosure may show an excellent effect even when solely used, but may be further used in combination with various methods such as hormone therapy, drug treatment, etc. to increase therapeutic efficiency.

The present disclosure may provide a method for preventing or treating idiopathic pulmonary fibrosis, including administering a compound represented by the above formula I, optical isomers thereof or pharmaceutically acceptable salts thereof into an individual.

Said terms "idiopathic pulmonary fibrosis," "prevention" and "treatment" may be the same as described above.

In the present disclosure, the term “administration” may refer to introducing a predetermined substance into an individual by an appropriate method.

In the present disclosure, the term “individual” may refer to all the animals such as rats, mice, livestock, etc., including humans, who have developed or are likely to develop idiopathic pulmonary fibrosis, and may be particularly mammals including humans, but is not limited thereto.

The method for preventing or treating idiopathic pulmonary fibrosis according to the embodiments of the present disclosure may include administering a therapeutically effective amount of the compound represented by the above formula I, optical isomers thereof or pharmaceutically acceptable salts thereof.

In the present disclosure, the term “therapeutically effective amount” may refer to an amount enough to treat a disease at a reasonable risk/benefit ratio applicable to medical treatment and not to cause a side effect, and may be determined by those skilled in the art according to factors including a patient’s gender, age, weight, health condition, a type of disease, severity, the activity of a drug, sensitivity to a drug, an administration method, an administration time, an administration route, an excretion rate, a treatment period, a drug combined or concurrently used, as well as other factors well known in a pharmaceutical field. It is preferable to differently apply a particular therapeutically effective amount for a certain patient depending on various factors including a type and degree of reaction to be achieved therefrom, a particular composition including a presence of other preparations used in some cases, a patient’s age, weight, general health condition, gender and diet, an administration time, an administration route, a secretion rate of the composition, a treatment period and a drug used together with the particular composition or simultaneously therewith, as well as other similar factors well known in a pharmaceutical field.

The method for preventing or treating idiopathic pulmonary fibrosis according to the embodiments of the present disclosure may include not only dealing with the disease per se before expression of its symptoms, but also inhibiting or avoiding such symptoms by administering the compound represented by the above formula I, isomers thereof or pharmaceutically acceptable salts thereof. In managing the disease, a preventive or therapeutic dose of a certain active ingredient may vary depending on the characteristics and severity of the disease or conditions, and a route in which the active ingredient is administered. A dose and a frequency thereof may vary depending on an individual patient’s age, weight and reactions. A suitable dose and usage may be easily selected by those skilled in the art, naturally considering such factors. In addition, the method for preventing or treating idiopathic pulmonary fibrosis according to the embodiments of the present disclosure may further include administering a therapeutically effective amount of an additional active agent, which helps prevent or treat the disease, along with the compound represented by the above formula I, optical isomers thereof or pharmaceutically acceptable salts thereof, and the additional active agent may show a synergy effect or an additive effect together with the compound represented by the above formula I, optical isomers thereof or pharmaceutically acceptable salts thereof.

The present disclosure may provide a use of the compound represented by the above formula I, optical isomers thereof or pharmaceutically acceptable salts thereof for preventing or treating idiopathic pulmonary fibrosis.

The present disclosure may provide a use of the compound represented by the above formula I, optical isomers thereof or pharmaceutically acceptable salts thereof in preparing a medicament for preventing or treating idiopathic pulmonary fibrosis.

For the preparation of the medicament, the compound represented by the above formula I, optical isomers thereof or pharmaceutically acceptable salts thereof may be mixed with pharmaceutically acceptable adjuvants, diluents, carriers, etc., and may be prepared into a complex preparation together with other active agents, thus providing a synergy action.

Matters mentioned in the pharmaceutical composition, treatment method and use of the present disclosure are applied the same, if not contradictory to each other.

Advantageous Effects

The compound represented by formula I of the present disclosure, optical isomers thereof or pharmaceutically acceptable salts thereof and the pharmaceutical composition including the same as an effective ingredient may be advantageously used in preventing or treating idiopathic pulmonary fibrosis.

Brief Description of the Drawings

FIG. 1 is a graph showing an expression level of fibrotic proteins (proCOLlAl and LOXL2) in each group. (* p < 0.05, ** p < 0.01, *** p < 0.001)

FIG. 2 is a graph showing a change in body weight of each group. (* p < 0.05)

FIG. 3 is a graph showing the number of total cells, neutrophils, lymphocytes, and macrophages in each group. (* p < 0.05)

FIG. 4 is a graph showing a lung weight of each group. (* p < 0.05)

FIG. 5 is a graph showing an expression level of fibrosis genes (COL1A1, FN1, TIMP1, PAI1 and SPP1) in each group. (* p < 0.05)

FIG. 6 is a graph showing an Ashcroft score of each group. (* p < 0.05)

FIG. 7 is a graph showing a degree of collagen 1A1(CO1A1) deposition in each group. (* p < 0.05)

FIG. 8 is an image showing a degree of collagen gel contraction in each group.

FIG. 9 is a graph showing a degree of collagen gel contraction in each group. (*** p < 0.001)

FIG. 10 is an image showing a degree of DHLF-IPF cell migration in each group.

FIG. 11 is a graph showing a degree of DHLF-IPF cell migration in each group. (* p < 0.05, ** p < 0.01)

FIG. 12 is a graph showing a degree of cell proliferation inhibition according to a concentration of the compound of the present invention. (** p < 0.01, *** p < 0.001)

Mode for Invention

The present disclosure will be described in detail with reference to Examples hereinafter. However, the Examples are only for the purpose of illustrating the present disclosure and it is obvious to those skilled in the art that the scope of the present disclosure is not limited to the Examples disclosed hereinafter.

Synthesis Example 1. Synthesis of N-(4-(hydroxycarbamoyl)benzyl)-N-(3-

(trifluoromethvI)phenvI)morphoIine-4-carboxamide [Compound 42]

[Step 1] Synthesis of methyl 4-((3-(trifluoromethyl)phenylamino)methyl)benzoate

A 3-(trifluoromethyl)benzenamine (0.30 g, 1.84 mmol) and potassium carbonate (0.76 g, 5.53 mmol) were dissolved in dimethylformamide (DMF, 5 mL), after which methyl 4-(bromomethyl)benzoate (0.42 g, 1.84 mmol) was inserted. A resulting solution was subjected to reaction at room temperature for a day and diluted with ethyl acetate. A reactant was washed with water and saturated sodium chloride aqueous solution, then dried and filtered with anhydrous magnesium sulfate, and then concentrated under reduced pressure. A residue was purified via column chromatography (silicon dioxide; ethyl acetate/hexane = 20%), such that a title compound (0.37 g, 65%) was obtained. 'H NMR (400 MHz, DMSO-de) 6 7.93 (d, 2 H, J = 8.3 Hz), 7.49 (d, 2 H, J = 8.3 Hz), 7.24 (t, 1 H, J = 7.9 Hz), 6.88-6.78 (m, 4 H), 4.42 (d, 2 H, J = 6.1 Hz), 3.83 (s, 3H), MS (ESI) m/z 310 (M⁺ + H).

[Step 2] Synthesis of methyl 4-((((4-nitrophenoxy)carbonyl)(3- (trifluoromethyl)phenyl)amino)methyl)benzoate

Methyl 4-((3-(trifluoromethyl)phenylamino)methyl)benzoate (0.26 g, 0.82 mmol) and 4-nitrophenyl carb onochlori date (0.33 g, 1.65 mmol) were dissolved in acetonitrile (10 mL), after which potassium carbonate (0.34 g, 2.47 mmol) was inserted. A resulting solution was subjected to reaction at room temperature for a day and diluted with ethyl acetate. A reactant was washed with saturated sodium chloride aqueous solution, then dried and filtered with anhydrous sodium sulfate, and then concentrated under reduced pressure. A residue was purified via column chromatography (silicon dioxide; ethyl acetate/hexane = 20%), such that a title compound (0.35 g, 89%) was obtained in a colorless oil form.

'H NMR (400 MHz, CDCh) 6 8.20 (d, 2 H, J = 10.2 Hz), 8.01 (d, 2 H, J = 7.8 Hz), 7.56-7.46 (m, 3H), 7.35 (d, 3 H, J = 8.0 Hz), 7.26 (d, 2 H, J = 8.1 Hz), 5.01 (bs, 2H), 3.90 (s, 3H).

[Step 3] Synthesis of methyl 4-((N-(3-(trifluoromethyl)phenyl)morpholine-4- carb oxami do)methy 1 )b enzoate

Methyl 4-((((4-nitrophenoxy)carbonyl)(3-

(trifluoromethyl)phenyl)amino)methyl)benzoate (0.29 g, 0.60 mmol) was dissolved in dimethylformamide (10 ml), after which potassium carbonate (0.25 g, 1.81 mmol) and morpholine (0.05 mL, 0.60 mmol) were inserted. A resulting solution was subjected to reaction at 60°C for two days, and then diluted with saturated ammonium chloride solution. An extraction was performed with ethyl acetate, after which a resulting extract was dried and filtered with anhydrous sodium sulfate, and then concentrated under reduced pressure. A residue was purified via column chromatography (silicon dioxide; ethyl acetate/hexane = 50%), such that a title compound (0.15 g, 60%) was obtained.

'H NMR (400 MHz, DMSO-de) 6 7.97 (d, 2 H, J = 8.2 Hz), 7.43-7.32 (m, 5H), 7.20 (d, 1 H, J = 8.0 Hz), 4.94 (s, 2H), 3.90 (s, 3H), 3.50 (t, 4 H, J = 4.8 Hz), 3.25 (t, 4 H, J = 4.8 Hz); MS (ESI) m/z 423 (M⁺ + H).

[Step 4] Synthesis of N-(4-(hydroxycarbamoyl)benzyl)-N-(3- (trifhioromethyl)phenyl)morpholine-4-carboxamide

Methyl 4-((N-(3-(trifluoromethyl)phenyl)morpholine-4- carboxamido)methyl)benzoate (0.15 g, 0.36 mmol) was dissolved in methanol (5 mL), after which hydroxylamine aqueous solution (50 wt%, 1 mL) and potassium hydroxide (0.10 g, 1.81 mmol) were inserted and stirred overnight. After a reaction was completed, a resulting solution was subjected to distillation under reduced pressure to remove methanol therefrom, after which an extraction was performed with ethyl acetate and water, such that work-up was done. A resulting extract was dried and filtered with anhydrous sodium sulfate, and then concentrated under reduced pressure. A residue was stirred in diethyl ether, after which a solid product was made, filtered and dried, such that a title compound (0.082 g, 54%) was obtained in a white solid form.

'H NMR (400 MHz, MeOD-d₃) 8 11.14 (brs, 1 H), 8.99 (brs, 1 H), 7.85 (d, 2 H, J = 8.0 Hz), 7.66-7.27 (m, 6 H), 4.94 (s, 2 H), 3.41 (s, 2 H), 3.15 (s, 2 H). MS (ESI) m/z 424 (M⁺ + H).

Synthesis Example 2. Synthesis of N-(2,4-difluoroDhenyl)-N-(4- i-4-carboxamide

[Step 1] Synthesis of methyl 4-((2,4-difhiorophenylamino)methyl)benzonate

A 2,4-difluorobenzeneamine (3.0 g, 23.2 mmol) and methyl 4-formylbenzoate (3.81 g, 23.2 mmol) were dissolved in methanol (500 mL) and stirred at room temperature for two hours, after which acetic acid (1.33 mL, 23.2 mmol) and sodium cyanoborohydride (1.46 g, 23.2 mmol) were added and stirred for a day. Methanol was slightly removed by air, after which a solid was precipitated, filtered and dried, so that a title compound (2.9 g, 45%) was obtained in a white solid form.

[Step 2] Synthesis of methyl 4-(((2,4-difluorophenyl)((4- nitrophenoxy)carbonyl)amino)methyl)benzoate

Methyl 4-((2,4-difluorophenylamido)methyl)benzoate (2 g, 7.21 mmol) and 4- nitrophenylchloroformate (1.45 g, 7.21 mmol) were dissolved in dichloromethane (50 mL) and stirred at room temperature for three days, after which water was added to extract an organic layer therefrom. The organic layer was washed with saturated sodium chloride aqueous solution, then dehydrated by anhydrous magnesium sulfate, and then concentrated under reduced pressure. A residue was dried, such that a title compound (2.5 g, 78%) was obtained in a yellow oil form.

[Step 3] Synthesis of methyl 4-((N-(2,4-difluorophenyl)morpholine-4- carb oxami do)methy l)b enzoate

Methyl 4-(((2,4-difluorophenyl)((4-nitrophenoxy)carbonyl)amino)methyl)benzoate (0.50 g, 1.13 mmol) and morpholine (0.098 mL, 1.13 mmol) were dissolved in dimethylformamide (10 mL) and heated and stirred at 60°C for two days. Dimethylformamide was removed under reduced pressure, after which water was poured into the reaction mixture and an extraction was performed with ethyl acetate. An organic layer was washed with saturated sodium chloride aqueous solution, then dehydrated by anhydrous magnesium sulfate, and then concentrated under reduced pressure. A residue was purified via column chromatography (silicon dioxide; ethyl acetate/hexane = 30%) and concentrated, so that a title compound (0.44 g, 98%) was obtained in a colorless oil form.

[Step 4] Synthesis of N-(2,4-difluorophenyl)-N-(4- (hydroxycarbamoyl)benzyl)morpholine -4-carboxamide

Methyl 4-((N-(2,4-difluorophenyl)morpholine-4-carboxamido)methyl)benzoate (0.10 g, 0.256 mmol) was dissolved in methanol (20 mL), after which hydroxylamine hydrochloride (0.089 g, 1.28 mmol) and potassium hydroxide (0.144 g, 2.56 mmol) were added and stirred, so that hydroxylamine (50 wt% aqueous solution; 0.329 mL, 5.12 mmol) was added dropwise and stirred at room temperature for three hours. After a reaction was completed, methanol was removed under reduced pressure, after which water was poured into the reaction mixture and an extraction was performed with ethyl acetate. An organic layer was washed with saturated sodium chloride aqueous solution, then dehydrated by anhydrous magnesium sulfate, and then concentrated under reduced pressure. After that, a resulting concentrate was dissolved in dichloromethane, after which hexane was added and a solid was precipitated, filtered and dried, so that a title compound (0.076 g, 76%) was obtained in a light yellow solid form.

'H NMR (400 MHz, MeOD-d₃) 6 7.65 (d, 2 H, J = 8.3 Hz), 7.41 (d, 2 H, J = 8.2 Hz), 7.27 - 7.25 (m, 1 H), 7.04 - 6.96 (m, 2 H), 4.80 (s, 2 H), 3.46 - 3.43 (m, 4 H), 3.22 - 3.19 (m, 4 H); MS (ESI) m/z 392.1 (M⁺ + H).

Synthesis Example 3. Synthesis of N-(4-(hydroxycarbamoyl)benzyl)-N-(3- methoxyphenyl)morpholine-4-carboxamide [Compound 104]

[Step 1] Synthesis of methyl 4-(((3-methoxyphenyl)amino)methyl)benzoate

An m-anisidine (3.23 g, 26.2 mmol) and methyl 4-(bromomethyl)benzoate (5.00 g, 21.8 mmol) were dissolved in acetonitrile (50 mL), after which N,N-diisopropylethylamine (5.80 mL, 32.7 mmol) was added and stirred at room temperature for 16 hours. When a reaction was completed, an extraction was performed with ethyl acetate and saturated sodium hydrogen carbonate aqueous solution, after which an organic layer was dried by anhydrous magnesium sulfate and filtered. A remaining filtrate was concentrated under reduced pressure, after which a residue was purified via column chromatography (silicon dioxide; ethyl acetate/hexane = 5%) and concentrated, such that a title compound (5.14 g, 87%) was obtained in a bright yellow liquid form.

[Step 2] Synthesis of methyl 4-(((3-methoxyphenyl)((4- nitrophenoxy)carbonyl)amino)methyl)benzoate

Methyl 4-(((3-methoxyphenyl)amino)methyl)benzoate (5.14 g, 18.9 mmol) and 4- nitrophenyl chloroformate (4.20 g, 20.8 mmol) were dissolved in acetonitrile (100 mL), after which potassium carbonate (3.93 g, 28.4 mmol) was added and stirred at room temperature for three hours. When a reaction was completed, an extraction was performed with ethyl acetate and saturated sodium hydrogen carbonate aqueous solution, after which an organic layer was dried by anhydrous magnesium sulfate and filtered. A remaining filtrate was concentrated under reduced pressure, after which a residue was purified via column chromatography (silicon dioxide; ethyl acetate/hexane = 20%) and concentrated, such that a title compound (5.88g, 71%) was obtained in a yellow liquid form.

[Step 3] Synthesis of methyl 4-((N-(3-methoxyphenyl)morpholine-4- carb oxami do)methy l)b enzoate

Methyl 4-(((3-methoxyphenyl)((4-nitrophenoxy)carbonyl)amino)methyl)benzoate (5.88 g, 13.5 mmol) was dissolved in dimethylformamide (50 mL), after which morpholine (2.35 g, 27.0 mmol) and potassium carbonate (5.60 g, 40.5 mmol) were added and stirred at 60°C for 16 hours. When a reaction was completed, an extraction was performed with ethyl acetate and saturated ammonium chloride aqueous solution, after which an organic layer was dried by anhydrous magnesium sulfate and filtered. A remaining filtrate was concentrated under reduced pressure, after which a residue was purified via column chromatography (silicon dioxide; ethyl acetate/hexane = 30%) and concentrated, such that a title compound (3.69g, 71%) was obtained in a yellow solid form.

[Step 4] Synthesis of N-(4-(hydroxycarbamoyl)benzyl)-N-(3- methyoxyphenyl)morpholine-4-carboxamide

Methyl 4-((N-(3-(methoxyphenyl)morpholine-4-carboxamido)methyl)benzoate (0.180 g, 0.468 mmol) was dissolved in methanol (10 mL), after which hydroxylamine (50.0 wt% aqueous solution; 1.43 mL, 23.4 mmol) was added at room temperature and then potassium hydroxide (0.263 g, 4.68 mmol) was added and stirred at the same temperature for 30 minutes. After that, a solvent was removed from the reaction mixture under reduced pressure. Saturated ammonium chloride aqueous solution was poured into the resulting concentrate, and an extraction was performed with ethyl acetate. An organic layer was washed with saturated sodium chloride aqueous solution, then dehydrated by anhydrous magnesium sulfate, and then concentrated under reduced pressure. A residue was crystallized at room temperature with di chloromethane (2 mL) and hexane (10 mL), such that a title compound (0.140 g, 78%) was obtained in a white solid form.

'H NMR (400 MHz, DMSO-de) 6 7.63 (d, 2 H, J = 8.1 Hz), 7.32 (m, 2 H), 7.19 (t, 1 H, J = 8.4 Hz), 6.69 - 6.67 (m, 2 H), 6.62 (m, 1 H), 4.84 (s, 2 H), 3.69 (s, 3 H), 3.39 - 3.36 (m, 4 H), 3.15 - 3.12 (m, 4 H). MS (ESI) m/z 386 (M⁺ + H).

Synthesis Example 4. Synthesis of N-(3-fluorophenyl)-N-(4- i-4-carboxamide

[Step 1] Synthesis of methyl 4-((3-fluorophenylamino)methyl)benzoate

Methyl 4-formylbenzoate (1.47 g, 8.99 mmol) was dissolved in methanol (50 mL), after which 3 -fluorobenzeneamine (1.0 g, 8.99 mmol) was inserted. A resulting solution was subjected to reaction at room temperature for three hours, after which sodium cyanoborohydride (NaCNBJL, 0.56 g, 8.99 mmol) and acetic acid (1.03 mL, 17.99 mmol) were inserted. A reactant was subjected to reaction at room temperature for a day, after which a reaction solvent was removed under reduced pressure, then saturated sodium hydrogen carbonate aqueous solution was poured, and then an extraction was performed by ethyl acetate. An organic layer was dehydrated by anhydrous magnesium sulfate, and concentrated under reduced pressure. A residue was purified via column chromatography (silicon dioxide; ethyl acetate/hexane = 20%), such that a title compound (1.84 g, 79%) was obtained.

[Step 2] Synthesis of methyl 4-(((3-fluorophenyl)((4- nitrophenoxy)carbonyl)amino)methyl)benzoate

Methyl 4-((3-fluorophenylamino)methyl)benzoate (2.7 g, 10.4 mmol) and 4- nitrophenyl chloroformate (4.20 g, 20.8 mmol) were dissolved in acetonitrile (100 mL), after which potassium carbonate (4.32 g, 31.2 mmol) was inserted. A resulting solution was subjected to reaction at room temperature for a day and diluted with ethyl acetate. A reactant was washed with saturated sodium chloride aqueous solution, then dried and filtered with anhydrous sodium sulfate, and then concentrated under reduced pressure. A residue was purified via column chromatography (silicon dioxide; ethyl acetate/hexane = 20%), such that a title compound (2.65 g, 60%) was obtained in a colorless oil form.

[Step 3] Synthesis of methyl 4-((N-(3-flurophenyl)morpholine-4- carb oxami do)methy 1 )b enzoate

Methyl 4-(((3-fluorophenyl)((4-nitrophenoxy)carbonyl)amino)methyl)benzoate

(0.32 g, 0.75 mmol) was dissolved in dimethylformamide (5 ml), after which potassium carbonate (0.31 g, 2.24 mmol) and morpholine (0.13 mL, 1.49 mmol) were inserted. A resulting solution was subjected to reaction at 60°C for a day, and then diluted with saturated ammonium chloride solution. An extraction was performed with ethyl acetate, after which a resulting extract was dried and filtered with anhydrous sodium sulfate, and then concentrated under reduced pressure. A residue was purified via column chromatography (silicon dioxide; ethyl acetate/hexane = 30%), such that a title compound (0.13 g, 45%) was obtained.

[Step 4] Synthesis of N-(3-fluorophenyl)-N-(4-

(hydroxycarbamoyl)benzyl)morpholine-4-carboxamide

Methyl 4-((N-(3-fluorophenyl)morpholine-4-carboxamido)methyl)benzoate (0.108 g, 0.290 mmol) was dissolved in methanol (10 mL), after which hydroxylamine (50.0 wt% aqueous solution; 1.19 mL, 19.4 mmol) was added at room temperature, and then potassium hydroxide (0.156 g, 2.78 mmol) was added and stirred at the same temperature for 16 hours. Then, a solvent was removed from the reaction mixture under reduced pressure, after which saturated sodium hydrogen carbonate aqueous solution was poured into the resulting concentrate and an extraction was performed with ethyl acetate. The organic layer was washed with saturated sodium chloride aqueous solution, then dehydrated by anhydrous magnesium sulfate, and then concentrated under reduced pressure. A precipitate solid was filtered and dried, such that a title compound (0.062 g, 57%) was obtained in a white solid form.

'H NMR (400 MHz, DMSO-de) 8 11.14 (brs, 1 H), 8.99 (brs, 1 H), 7.65 (d, 2 H, J = 7.0 Hz), 7.38-7.30 (m, 3H), 7.05-6.85 (m, 3H), 4.89 (s, 1H), 3.44-3.42 (m, 4H), 3.18-3.15 (m, 4H), 2.08 (s, 3H). MS (ESI) m/z 374 (M⁺ + H).

Synthesis Example 5. Synthesis of N-(3-fluorophenyl)-N-(4-

[Step 1] Synthesis of methyl 4-((N-(3 -fluorophenyl)- l,4-oxazepane-4- carb oxami do)methy l)b enzoate

Methyl 4-(((3-fluorophenyl)((4-nitrophenoxy)carbonyl)amino)methyl)benzoate (0.290 g, 0.683 mmol) obtained in step 2 of Preparation Example 5, 1,4-oxazepane (0.188 g, 1.367 mmol) and potassium carbonate (0.283 g, 2.050 mmol) were dissolved in DMF (10 mL), after which the reaction solution was stirred at 60°C for a day, then saturated NaHCOs aqueous solution was poured into the reaction mixture, and then an extraction was performed with ethyl acetate. An organic layer was washed with saturated sodium chloride aqueous solution, then dehydrated by anhydrous magnesium sulfate, and then concentrated under reduced pressure. A resulting concentrate was purified via column chromatography (silicon dioxide, 15 g cartridge; ethyl acetate/hexane = 20 to 50%) and concentrated, such that a title compound (0.116 g, 43.9%) was obtained in a colorless liquid form.

[Step 2] Synthesis of N-(3-fluorophenyl)-N-(4-hydroxy carbarn oyl)benzyl)- 1,4- oxazepane-4-carboxamide

Methyl 4-((N-(3 -(fluorophenyl)- l,4-oxazepane-4-carboxamide)methyl)benzoate (0.116 g, 0.3 mmol) was dissolved in methanol (10 mL), after which hydroxylamine aqueous solution (50 wt%, 1 mL) and potassium hydroxide (0.168 g, 3.01 mmol) were inserted and stirred overnight. After a reaction was completed, a resulting solution was subjected to distillation under reduced pressure to remove methanol therefrom, after which an extraction was performed with ethyl acetate and water, such that work-up was done. A resulting extract was dried and filtered with anhydrous sodium sulfate and concentrated under reduced pressure, after which a residue was stirred in diethyl ether, such that a solid product was made, filtered and dried, and thus a title compound (0.032 g, 27.5%) was obtained in a white solid form. <Example 1> Effect of inhibiting expression of fibrotic proteins

In order to confirm an effect of the compound according to the present disclosure on preventing or treating idiopathic pulmonary fibrosis, an expression of fibrotic proteins was analyzed.

MRC-5 cells, a lung fibroblast cell line, were seeded at 1 x 10⁵ cells/well in a 6- well plate and cultured in a CO2 incubator (37°C, 5% CO2) for 24 hours. After that, for serum starvation, a culture medium was replaced with an EMEM (1% FBS, 1% P/S) culture medium and cultured in a CO2 incubator (37°C, 5% CO2) for 24 hours. After the serum starvation, 1 pM of test substances (42, 68, 104, 130 and 151 compounds) were treated with 5 ng/mL of TGF-pi and cultured in a CO2 incubator (37°C, 5% CO2) for 48 hours. A culture medium containing 0.1% DMSO was added to a control group and a TGF-pi only treated group. The expression of fibrotic proteins (proCOLlAl, LOXL2) in the completely cultured cells was compared by using western blot.

Cells were lysed in 100 pL of RIPA buffer (containing proteinase & phosphatase inhibitors) and incubated on ice for 30 minutes. The resulting product was centrifuged at 13,000 g, 4°C for 20 minutes. The supernatant was separated and quantified by using a BCA protein assay kit. Then, a NuPAGE sample reducing agent and a NuPAGE LDS sample buffer (4X) were added to prepare a sample at a concentration of 0.5 pg/pL. The prepared sample was boiled at 100°C for five minutes to denaturate the proteins. 5 pg of protein was separated at 120 V after loading on NuPAGE Novex 4-12% Bis-Tris gel and transferred to a nitrocellulose (NC) membrane through an iBlot 2 dry blotting system. After that, the membrane was blocked with a blocking solution (EzBlock Chemi: distilled water = 1 : 4) at room temperature for 30 minutes. The membrane was reacted overnight with a primary antibody at 4°C, and then washed three times with l x TBST for 10 minutes each. Then, the resulting product was reacted with a horseradish peroxidase (HRP)-linked secondary antibody at room temperature for one hour, and washed three times with l x TBST for 10 minutes each. The primary and secondary antibodies were diluted in a solution of a blocking solution and l x TBST mixed in a ratio of 1 :4. After that, the proteins were visualized by using Amersham™ ECL select™ western blotting detection reagent and ChemiDoc™ MP imaging system. The observed proteins were quantified by using Image Lab 5.0 software, and then corrected by using a P-actin level.

One-way ANOVA was performed by using GraphPad Prism 5.0 software (post hoc: Dunnett’s multiple comparison test). All data are expressed as mean±SEM, and P<0.05 was considered statistically significant.

As a result, as shown in FIG. 1, it was confirmed that the expression of proCOLlAl and LOXL2 induced by TGF-pi is remarkably inhibited when treated with the compound of the present disclosure.

Thus, it could be seen that the compound of the present disclosure shows an effect of inhibiting an expression of fibrotic proteins, and thus may be advantageously used in preventing or treating idiopathic pulmonary fibrosis.

<Example 2> Therapeutic effect in animal model with BLM-induced pulmonary fibrosis

Eight-week-old male C57BL/6 mice were anesthetized by intraperitoneal (i.p.) administration of ketamine hydrochloride (2 mg/mouse) and xylazine hydrochloride (0.07 mg/mouse), and then intranasally (i.n.) dosed with a bleomycin (BLM) solution at a concentration of 1 mg/kg (50 pL/mouse), so as to prepare a pulmonary fibrosis model. A test substance was administered for a total of 14 days from one week after administration of the BLM.

In order to confirm a therapeutic effect of the compound of the present disclosure, the mice with BLM-induced pulmonary fibrosis were divided into groups as shown in table 1 below according to an administered substance [vehicle (Veh) or compound 42 (42)], a route of administration [oral administration (PO)], and an administration interval [once daily (QD)].

[Table 11

Two-way ANOVA was performed by using GraphPad Prism 8.1.1 software (Mann- Whitney test). All data are expressed as mean±SEM, and P<0.05 was considered statistically significant.

<Example 2-l> Analysis of weight changes In order to confirm an effect of the compound according to the present disclosure on preventing or treating idiopathic pulmonary fibrosis, a change in body weights of mice was analyzed. Body weights were measured and recorded daily.

As a result, as shown in FIG. 2, it was confirmed that a weight loss appears in mice with BLM-induced pulmonary fibrosis and the weight loss is recovered upon treatment with the compound of the present disclosure.

Thus, it could be seen that the compound of the present disclosure shows an effect of recovering a reduced weight, and thus may be advantageously used in preventing or treating idiopathic pulmonary fibrosis.

<Example 2-2> Analysis of bronchoalveolar lavage fluid (BALF)

In order to confirm an effect of the compound according to the present disclosure on preventing or treating idiopathic pulmonary fibrosis, bronchoalveolar lavage fluid (BALF) was analyzed. After inserting a cannula into the trachea to collect the BALF, the lungs were washed with PBS (0.4, 0.3 and 0.3 mL, in a total of 1 mL). The collected BALF was placed in an Eppendorf test tube, centrifuged (5 min/3500 rpm/4°C), and resuspended in 600 pL of PBS. The number of total cells and differentiated cells was analyzed.

As a result, as shown in FIG. 3, it was confirmed that the number of total cells, neutrophils, lymphocytes, and macrophages is increased in mice with BLM-induced pulmonary fibrosis, and then the increased number of total cells, neutrophils, lymphocytes and macrophages is decreased when treated with the compound of the present disclosure.

Thus, it could be seen that the compound of the present disclosure shows an effect of inhibiting the infiltration of inflammatory cells, and thus may be advantageously used in preventing or treating idiopathic pulmonary fibrosis.

<Example 2-3> Analysis of lung weight changes

In order to confirm an effect of the compound according to the present disclosure on preventing or treating idiopathic pulmonary fibrosis, a change in lung weights of mice was analyzed. The weight of the lungs removed from the mice was measured.

As a result, as shown in FIG. 4, it was confirmed that a weight of lungs is increased in mice with BLM-induced pulmonary fibrosis and an increased weight of lungs is decreased upon treatment with the compound of the present disclosure.

Thus, it could be seen that the compound of the present disclosure shows an effect of decreasing an increased lung weight, and thus may be advantageously used in preventing or treating idiopathic pulmonary fibrosis.

<Example 2-4> Analysis of gene expression

In order to confirm an effect of the compound according to the present disclosure on preventing or treating idiopathic pulmonary fibrosis, an expression of fibrosis genes was analyzed. Lung tissue was placed in a Precelllys CK28 Hard Tissue tube containing 750 pL of trizol reagent and homogenized. 150 pL of chloroform was added to the homogenate, shaken vigorously, and then incubated at room temperature for three minutes, followed by centrifugation at 6,000 g, 4°C for 15 minutes for phase separation. An aqueous phase was collected and RNA was further isolated by using the NucleoSpin 96 RNA Core kit according to the manufacturer's instructions (Macherey -Nagel). RNA was eluted in 60 pL of RNase free water, and an RNA concentration was determined by using absorbance at 260 nm. A total of 500 ng of isolated RNA was reverse transcribed into cDNA by using the High Capacity RT Kit according to the manufacturer's instructions (Life Technologies). After completion of a reverse transcription reaction, the obtained cDNA was diluted five-fold in RNAse free water. The gene expression of COK1A1, FN1, PAI1, TIMP-1 and SPP1 was measured by using real time PCR AriaMX Instrument (Agilent Technologies).

As a result, as shown in FIG. 5, it was confirmed that an expression of COK1A1, FN1, PAI1, TIMP-1 and SPP1 genes is increased in mice with BLM-induced pulmonary fibrosis and an increased expression of the genes is decreased upon treatment with the compound of the present disclosure.

Thus, it could be seen that the compound of the present disclosure shows an effect of inhibiting an expression of fibrosis genes, and thus may be advantageously used in preventing or treating idiopathic pulmonary fibrosis.

<Example 2-5> Histopathologic analysis

In order to confirm an effect of the compound according to the present disclosure on preventing or treating idiopathic pulmonary fibrosis, a histopathologic evaluation was performed. A lung specimen was embedded in paraffin. Slides were stained for fibrous tissue. A histological change in the lungs was evaluated by using the Ashcroft score. A de novo collagen 1A1(CO1A1) deposition was evaluated by digital image analysis (Calopix software, TRIBVN). As a result, as shown in FIG. 6, it was confirmed that an Ashcroft score is increased in mice with BLM-induced pulmonary fibrosis and an increased Ashcroft score is decreased upon treatment with the compound of the present disclosure.

In addition, as shown in FIG. 7, it was confirmed that the de novo collagen 1A1 deposition is increased in mice with BLM-induced pulmonary fibrosis and the increased de novo collagen 1 Al deposition is decreased upon treatment with the compound of the present disclosure.

Thus, it could be seen that the compound of the present disclosure shows an anti- fibrotic effect, and thus may be advantageously used in preventing or treating idiopathic pulmonary fibrosis.

<Example 3> Collagen gel contraction assay

In order to confirm an effect of the compound according to the present disclosure on preventing or treating idiopathic pulmonary fibrosis, a degree of collagen gel contraction was analyzed.

3 ng/mL of TGF-pi was treated alone or with compound 42 at various concentrations (0.3, 1, 3, 10 pM) to a collagen gel containing 4 x 10⁵ LL29 cells (pulmonary fibrosis cells derived from an IPF patient), and incubated for 192 hours. A control group and a TGF-pi only treated group were treated with 0.1% DMSO.

As a result, as shown in FIGS. 8 and 9, it was confirmed that a collagen contraction induced by TGF-pi is remarkably inhibited when treated with the compound of the present disclosure.

<Example 4> Fibroblast migration assay

In order to confirm an effect of the compound according to the present disclosure on preventing or treating idiopathic pulmonary fibrosis, a fibroblast migration was analyzed.

A diseased human lung fibroblast-IPF (DHLF-IPF) derived from lung tissue of an IPF patient was cultured as a confluent monolayer. After 24 hours of serum starvation, scratches were made by using a 200 pL micropipette tip. After that, compound 42 was prepared and treated in a cell culture medium to a final concentration of 1.5 and 3 pM. A control group was treated with 0.1% DMSO. After further culturing for 15 hours, a cell migration distance was measured by using a phase-contrast microscope.

As a result, as shown in FIGS. 10 and 11, it was confirmed that a migration of DHLF-IPF cells is remarkably inhibited when treated with the compound of the present disclosure.

<Example 5> Fibroblast proliferation assay

In order to confirm an effect of the compound according to the present disclosure on preventing or treating idiopathic pulmonary fibrosis, a fibroblast proliferation was analyzed.

Normal human lung fibroblast (NHLF) cells labeled with eFluor670 dye were seeded at 8 * 10⁵ cells in a 100 mm dish (Day 0). In 24 hours after seeding (Day 1), 30 ng/mL of PDGF-BB was treated alone or with compound 42 at various concentrations (0.01, 0.03, 0.3, 1, and 3 pM), and then cultured for 48 hours (Day 3). On Day 3, the cells were trypsinized and suspended into single cells, and cell proliferation was evaluated by using flow cytometry.

As a result, as shown in FIG. 12, it was confirmed that a proliferation of cells induced by PDGF-BB is inhibited when treated with the compound of the present disclosure.

Thus, it could be seen that the compound of the present disclosure shows an anti- fibrotic effect, and thus may be advantageously used in preventing or treating idiopathic pulmonary fibrosis. The present disclosure provides a pharmaceutical composition, a method, and a use as follow:

Item 1. A pharmaceutical composition for preventing or treating idiopathic pulmonary fibrosis, comprising a compound represented by the above-mentioned formula I, optical isomers thereof or pharmaceutically acceptable salts thereof as an active ingredient.

Item 2. The pharmaceutical composition of item 1, wherein the compound represented by the above-mentioned formula I is at least one selected from the group consisting of the above-mentioned compounds 1 to 169 which is described in the above- mentioned Table.

Item 3. The pharmaceutical composition of item 1 or 2, wherein the compound represented by the above-mentioned formula I is at least one selected from the group consisting of compound 42, compound 68, compound 104, compound 130 and compound 151 which is described in the above-mentioned Table.

Item 4. A method for preventing or treating idiopathic pulmonary fibrosis, including administering the compound represented by formula I, optical isomers thereof or pharmaceutically acceptable salts thereof described in any one of items 1 to 3 into an individual.

Item 5. A use of the compound represented by formula I, optical isomers thereof or pharmaceutically acceptable salts thereof described in any one of items 1 to 3 for preventing or treating idiopathic pulmonary fibrosis.

Item 6. A use of the compound represented by formula I, optical isomers thereof or pharmaceutically acceptable salts thereof described in any one of items 1 to 3 in preparing a medicament for preventing or treating idiopathic pulmonary fibrosis.

Item 7. The pharmaceutical composition according to any one of items 1 to 3, wherein the pharmaceutical composition is orally administered.

Item 8. The method according to item 4, or the use according to item 5 or 6, wherein the compound represented by formula I, optical isomers thereof or pharmaceutically acceptable salts thereof described in any one of items 1 to 3 is orally administered.

While specific portions of the present disclosure have been described in detail above, it is apparent to those skilled in the art that such detailed descriptions are set forth to illustrate exemplary embodiments only, but are not construed to limit the scope of the present disclosure. Thus, it should be understood that the substantial scope of the present disclosure is defined by the accompanying claims and equivalents thereto.

Claims

Claim s

1. A pharmaceutical composition for preventing or treating idiopathic pulmonary fibrosis, comprising a compound represented by a following formula I, optical isomers thereof or pharmaceutically acceptable salts thereof as an effective ingredient:

[Formula I]

Xa and Xb are each independently CH or N,

Q is C(=O), S(=O)₂, S(=O) or C(=NH),

Y is selected from a following group:

M is C, N, O, S or S(=O)₂ (at this time, if M is C, 1 and m are 1; if M is N, 1 is 1 and m is 0; and if M is O, S or S(=O)2, 1 and m are 0),

Rai and Ra2 are each independently hydrogen; hydroxy; -Ci-4 straight or branched chain alkyl, which is unsubstituted or substituted with at least one halogen; -Ci-4 straight or branched chain alcohol; benzhydryl; -Ci-4 straight or branched chain alkyl, which is substituted with a saturated or unsaturated five to seven-membered heterocyclic compound

45 having one to three heteroatoms out of N, O or S as a ring member (at this time, the heterocyclic compound may be unsubstituted or at least one hydrogen may be optionally substituted with OH, OCH3, CH3, CH2CH3 or halogen); a saturated or unsaturated five to seven-membered heterocyclic compound having one to three heteroatoms out of N, O or S as a ring member (at this time, the heterocyclic compound may be unsubstituted or at least one hydrogen may be optionally substituted with OH, OCH3, CH3, CH2CH3 or halogen); phenyl, which is unsubstituted or in which at least one hydrogen is substituted with halogen, Ci-4 alkoxy, C1-2 alkyl or hydroxy; benzyl, which is unsubstituted or in which at least one hydrogen is substituted with halogen, Ci-4 alkoxy, C1-2 alkyl or hydroxy; -S(=O)2CH3; halogen; -C1-6 straight or branched chain alkoxy; -C2-6 alkoxyalkyl; -C(=O)R_x, in which R_x is

C1-3 straight or branched chain alkyl or C3-10 cycloalkyl;

n is an integer of 0, 1 or 2,

Z is selected from a following group:

46

Pa and Pb are each independently

hydrogen; hydroxy; -Ci-4 straight or branched chain alkyl, which is unsubstituted or in which at least one hydrogen is substituted with halogen; halogen; -CF3; -OCF3; -CN; -C1-6 straight or branched chain alkoxy; -C2-6 straight or branched chain alkyl alkoxy; -CH2F; or -C1-3 alcohol,

( B ! in which ' is a ring selected from phenyl, pyridine, pyrimidine, thiazole, indole, indazole, piperazine, quinoline, furan, tetrahydropyridine, piperidine or a following group:

x, y and z are each independently an integer of 0 or 1, and

Rgi, R_g2 and R_g3 are each independently selected from hydrogen; hydroxy; -C1-3 alkyl; -CF3; -C1-6 straight or branched chain alkoxy; -C2-6 straight or branched chain alkyl alkoxy; -C(=O)CH3; -Ci-4 straight or branched chain hydroxyalkyl; -N(CH3)2; halogen; phenyl; -S((=O)2)CH3; or a following group:

2. The pharmaceutical composition according to claim 1, wherein the compound represented by formula l is a compound represented by a following formula la:

47

wherein

Y is selected from a following group:

in which M, 1, m, n, Rai, Ra2, and Rb are the same as defined in claim 1, respectively,

3. The pharmaceutical composition according to claim 2, wherein Y is selected from a following group:

in which n and Rb are the same as defined in claim 1, respectively,

Pa and Pb are each independently hydrogen; halogen; -CF3; or -C1-6 straight or branched chain alkoxy.

4. The pharmaceutical composition according to claim 1, wherein the compound represented by formula l is a compound described in a following table:

5. The pharmaceutical composition according to claim 1, wherein the compound represented by formula I is a compound described in a following table:

6. The pharmaceutical composition according to claim 1, wherein the pharmaceutical composition is orally administered.

7. A method for preventing or treating idiopathic pulmonary fibrosis, comprising administering a compound represented by formula I, optical isomers thereof or pharmaceutically acceptable salts thereof into an individual, wherein the compound represented by formula I is the same as defined in claim 1.

8. The method according to claim 7, wherein the compound represented by formula l is compound described in a following table:

62

9. A use of a compound represented by formula I, optical isomers thereof or pharmaceutically acceptable salts thereof for preventing or treating idiopathic pulmonary fibrosis, wherein the compound represented by formula I is the same as defined in claim 1.

10. The use according to claim 9, wherein the compound represented by formula l is compound described in a following table:

11. A use of a compound represented by formula I, optical isomers thereof or pharmaceutically acceptable salts thereof in preparing a medicament for preventing or treating idiopathic pulmonary fibrosis, wherein the compound represented by formula I is the same as defined in claim 1.

12. The use according to claim 11, wherein the compound represented by formula I is a compound described in a following table:

64