KR102563719B1 - Pharmaceutical composition for preventing or treating IFTA acting as a functional inhibitor for S1PR1 and S1PR4 - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for preventing or treating IFTA, which acts as a functional inhibitor for S1P receptor subtypes S1PR1 and S1PR4, and more specifically, acts as a functional inhibitor for S1PR1 and S1PR4 and does not cause side effects of cardiovascular disease, It relates to a pharmaceutical composition comprising a sphingolipid compound having an IFTA preventive or therapeutic effect as an active ingredient.

Description

Pharmaceutical composition for preventing or treating IFTA acting as a functional inhibitor for S1PR1 and S1PR4

The present invention relates to a composition for preventing or treating IFTA (Interstitial Fibrosis and Tubular Atrophy, interstitial fibrosis and tubular atrophy) acting as a functional inhibitor for S1PR1 and S1PR4, and more specifically, the present invention is a functional inhibitor for S1PR1 and S1PR4. The present invention relates to a pharmaceutical composition containing a sphingolipid compound that does not cause side effects of cardiovascular disease and has an effect of preventing or treating Interstitial Fibrosis and Tubular Atrophy (IFTA).

The number of organ transplants worldwide increased by 5.8% from the previous year to about 120,700 in 2015, of which about 60% are kidney transplants. IFTA (Interstitial Fibrosis and Tubular Atrophy) develops in about 25% of patients 1 year after kidney transplantation. As the interstitial space expands and replaces normal tissue due to histopathological changes in the transplanted kidney, tubular atrophy and interstitial fibrosis occur, resulting in decreased glomerular filtration rate and increased proteinuria. 66% in the 5th year after transplantation and up to 90% in the 10th year after transplantation, these symptoms are mainly caused by immunological causes in the early stages, but in the later stages are mainly due to the toxicity of calcineurin inhibitors, which are immunosuppressive agents.

Although the mechanism of IFTA induction of calcineurin inhibitors, which are mainly used for post-transplantation immunosuppressive maintenance therapy, is not clear, it provides the cause of fibrosis by inducing vasoconstriction, oxidative stress, and TGF-β. It is known.

Currently, calcineurin inhibitors, anti-metabolites, steroid combination therapy, etc. are generally used for IFTA. If developed, it is expected to improve the quality of life of patients after transplantation.

Accordingly, there is a continuous demand for the development of drugs that can be used for IFTA treatment and have excellent drug efficacy and are safe in terms of side effects or toxicity.

Accordingly, while the inventors of the present invention made intensive efforts to develop an independent compound that selectively acts on the S1P subtype receptor among sphingolipid compounds, the compound according to the present invention specifically binds to the S1PR1 and S1PR4 receptors and acts as a functional inhibitor (antagonist). ), the present invention was completed by confirming that there is an IFTA preventive or therapeutic effect without causing side effects of cardiovascular diseases due to non-selection for the S1PR subtype.

Korean Patent Publication No. 10-2017-0087813 (2017.07.31)

The present invention provides a pharmaceutical composition containing, as an active ingredient, a compound having pharmacological activity for preventing or treating IFTA (Interstitial Fibrosis and Tubular Atrophy) without causing side effects of cardiovascular disease.

here,

R ¹ is hydrogen;

R ² is hydrogen or an acetyl group;

X is a single bond, C ₂ alkylene, or C ₂ alkenylene;

A is a 5-membered ring heteroarylene containing 3 N atoms;

B is C _2-11 straight or branched chain alkylene;

C is a single bond or phenylene; and

D is selected from the group consisting of hydrogen, phenyl and C _1-6 alkyl.

The R ² may be hydrogen.

The X may be C ₂ alkylene.

The B may be C _2-11 straight chain alkylene.

The compound represented by Formula 1 may be any one compound selected from the group of compounds below.

(1) 2-amino-2-(2-(1-decyl-1H-1,2,3-triazol-4-yl)ethyl)propane-1,3-diol;

(2) 2-amino-2-(2-(1-octyl-1H-1,2,3-triazol-4-yl)ethyl)propane-1,3-diol;

(3) 2-amino-2-(2-(1-(4-hexylphenethyl)-1H-1,2,3-triazol-4-yl)ethyl)propane-1,3-diol;

(4) 2-amino-2-(1-dodecyl-1H-1,2,3-triazol-4-yl)propane-1,3-diol;

(5) (E)-2-amino-2-(1-decyl-1H-1,2,3-triazol-4-yl)vinyl-1,3-diol;

(6) 2-amino-2-(2-(1-(8-phenyloctyl)-1H-1,2,3-triazolebutyl-4-yl)ethyl)propane-1,3-diol;

(7) N-(2-(1-dodecyl-1H-1,2,3-triazol-4-yl)-1,3-dihydrooxypropan-2-yl)acetamide;

(8) N-(4-(1-decyl-1H-1,2,3-triazol-4-yl)-1-hydroxy-2-(hydroxymethyl)butan-2-yl)acetamide; and

(9) N-(4-(1-(4-hexylphenethyl)-1H-1,2,3-triazol-4-yl)-1-hydroxy-2-(hydroxymethyl)butane-2 -yl) acetamide

The compound represented by Chemical Formula 1 may be a compound represented by Chemical Formula 2 (hereinafter referred to as 'NXC736').

[Formula 2]

The pharmaceutical composition according to the present invention may be a preparation for oral administration or a preparation for parenteral administration.

A pharmaceutical composition according to the present invention may further comprise one or more other therapeutic agents suitable for the treatment of IFTA.

The pharmaceutical composition according to the present invention can act as a functional inhibitor for S1PR1 and S1PR4.

The pharmaceutical composition according to the present invention can be expected to have effects that do not cause cardiovascular disease side effects.

The pharmaceutical composition according to the present invention acts as a functional inhibitor for S1PR1 and S1PR4 and has an effect of preventing or treating IFTA.

In particular, the pharmaceutical composition according to the present invention acts as a functional inhibitor for S1PR1 and S1PR4 among subtypes of S1P receptors (S1P1, S1P2, S1P3, S1P4 and S1P5), thereby preventing or treating IFTA and at the same time reducing cardiovascular disease side effects. There is an effect that does not cause.

1 is a graph showing changes in the number of total T cells and T cell subtypes in mice after oral administration of NXC736 (3 mg/kg/day).
Figure 2 is a graph showing changes in the number of total T cells and T cell subtypes in rats after oral administration of NXC736 (10 mg/kg/day).
Figure 3 is a graph showing the effect of NXC736 (10, 30, 90mg / kg / day) on renal fibrosis in the UUO mouse model.
Figure 4 is a table evaluating the effect of NXC736 (1, 3, 10, 30 μM) on HEK293 cells in which the hERG gene was overexpressed.
5 is a graph showing the heart rate measurement results of beagle dogs after oral administration of NXC736 (12.5, 25, 50 mg/kg).

The descriptions disclosed in this specification or application are merely exemplified for the purpose of explaining embodiments according to the technical spirit of the present invention, and embodiments according to the technical spirit of the present invention are the embodiments disclosed in this specification or application. In addition to these, it may be implemented in various forms, and the technical spirit of the present invention is not construed as being limited to the embodiments described in this specification or application.

In the present invention, "prevention" refers to any action that suppresses or delays the onset of a target disease by administering a pharmaceutical composition to a subject.

"Treatment" in the present invention refers to any action that improves or benefits the symptoms of a target disease by administering a pharmaceutical composition to a subject.

Hereinafter, the present invention will be described in detail.

The pharmaceutical composition for the prevention or treatment of IFTA (Interstitial Fibrosis and Tubular Atrophy) according to the present invention contains a compound represented by Formula 1 below, an optical isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

[Formula 1]

here,

R ¹ is hydrogen;

R ² is hydrogen or an acetyl group;

X is a single bond, C ₂ alkylene, or C ₂ alkenylene;

A is a 5-membered ring heteroarylene containing 3 N atoms;

B is C _2-11 straight or branched chain alkylene;

C is a single bond or phenylene; and

D is selected from the group consisting of hydrogen, phenyl and C _1-6 alkyl.

The R ² may be hydrogen.

The X may be C ₂ alkylene.

The B may be C _2-11 straight chain alkylene.

In addition, the present invention provides a method for preventing or treating IFTA comprising administering to a subject a material selected from the compound represented by Formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof. The subject may be a human or non-human animal.

The substance may act as a functional inhibitor for S1PR1 and S1PR4 to provide a method for preventing or treating IFTA that does not cause cardiovascular disease side effects.

In addition, the present invention provides a composition containing as an active ingredient a material selected from the compound represented by Formula 1, its optical isomer or its pharmaceutically acceptable salt, for use in preventing or treating IFTA.

The composition acts as a functional inhibitor for S1PR1 and S1PR4, and can provide IFTA preventive or therapeutic uses that do not cause cardiovascular disease side effects.

In addition, the compound represented by Formula 1 may be any one compound selected from the following compound groups.

(1) 2-amino-2-(2-(1-decyl-1H-1,2,3-triazol-4-yl)ethyl)propane-1,3-diol;

(2) 2-amino-2-(2-(1-octyl-1H-1,2,3-triazol-4-yl)ethyl)propane-1,3-diol;

(3) 2-amino-2-(2-(1-(4-hexylphenethyl)-1H-1,2,3-triazol-4-yl)ethyl)propane-1,3-diol;

(4) 2-amino-2-(1-dodecyl-1H-1,2,3-triazol-4-yl)propane-1,3-diol;

(5) (E)-2-amino-2-(1-decyl-1H-1,2,3-triazol-4-yl)vinyl-1,3-diol;

(6) 2-amino-2-(2-(1-(8-phenyloctyl)-1H-1,2,3-triazolebutyl-4-yl)ethyl)propane-1,3-diol;

(7) N-(2-(1-dodecyl-1H-1,2,3-triazol-4-yl)-1,3-dihydrooxypropan-2-yl)acetamide;

(8) N-(4-(1-decyl-1H-1,2,3-triazol-4-yl)-1-hydroxy-2-(hydroxymethyl)butan-2-yl)acetamide; and

(9) N-(4-(1-(4-hexylphenethyl)-1H-1,2,3-triazol-4-yl)-1-hydroxy-2-(hydroxymethyl)butane-2 -yl) acetamide

In addition, the compound represented by Formula 1 is 2-amino-2-(2-(1-decyl-1H-1,2,3-triazol-4-yl)ethyl)propane, which is a compound represented by Formula 2 below. -1,3-diol.

[Formula 2]

The compound represented by Formula 1 may be used in the form of a pharmaceutically acceptable salt. In this case, an acid addition salt formed by a pharmaceutically acceptable free acid may be used as the salt.

More specifically, acid addition salts are inorganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, nitrous acid, phosphorous acid, etc., aliphatic mono- and dicarboxylates, phenyl-substituted alkanoates, hydroxyl From organic acids such as acetic acid, benzoic acid, citric acid, lactic acid, maleic acid, gluconic acid, methanesulfonic acid, 4-toluenesulfonic acid, tartaric acid, fumaric acid, etc. can be obtained.

In addition, pharmaceutically non-toxic salts include sulfate, pyrosulfate, bisulphate, sulfite, bisulfite, nitrate, phosphate, monohydrogen phosphate, dihydrogen phosphate, metaphosphate, pyrophosphate chloride, bromide, ioda Id, fluoride, acetate, propionate, decanoate, caprylate, acrylate, formate, isobutyrate, caprate, heptanoate, propiolate, oxalate, malonate, succinate, suberate , sebacate, fumarate, maleate, butyne-1,4-dioate, hexane-1,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitro benzoate, hydroxybenzoate, Toxybenzoate, phthalate, terephthalate, benzenesulfonate, toluenesulfonate, chlorobenzenesulfonate, xylenesulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, β-hydroxybutyrate, glycol rate, maleate, tartrate, methanesulfonate, propanesulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate, mandelate, and the like.

Acid addition salts can be prepared by conventional methods. For example, it is prepared by dissolving a derivative of the compound represented by Formula 1 in an organic solvent such as methanol, ethanol, acetone, methylene chloride, acetonitrile, etc., and adding an organic acid or inorganic acid to filter and dry the resulting precipitate, or It can be prepared by distilling an acid under reduced pressure, drying it, and crystallizing it in an organic solvent.

In addition, a pharmaceutically acceptable metal salt may be prepared using a base. For example, it can be obtained by dissolving the compound represented by formula (1) in an excess alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the undissolved compound salt, and evaporating and drying the filtrate. As the metal salt, it is preferable to prepare a sodium, potassium or calcium salt. Corresponding salts may also be obtained by reacting an alkali metal or alkaline earth metal salt with a suitable silver salt (eg, silver nitrate).

In addition, the pharmaceutical composition according to the present invention not only contains the compound represented by Formula 1 and its pharmaceutically acceptable salt as an active ingredient, but also contains a substance selected from solvates, optical isomers and hydrates that can be prepared therefrom. It can be included as an active ingredient.

A pharmaceutical composition for the prevention or treatment of IFTA containing the compound represented by Formula 1, its optical isomer or its pharmaceutically acceptable salt as an active ingredient is a pharmaceutical composition for oral administration and other types of preparations within the range of pharmacological activity. can be changed

Formulations for oral administration may be in the form of troches, lozenges, tablets, aqueous suspensions, oily suspensions, prepared powders, granules, emulsions, hard capsules, soft capsules, syrups or elixirs, It is not limited to this.

In addition, in order to formulate the pharmaceutical composition according to the present invention into a preparation for oral administration, a binder such as lactose, saccharose, sorbitol, mannitol, starch, amylopectin, cellulose or gelatin; excipients such as dicalcium phosphate; disintegrants such as corn starch or sweet potato starch; lubricants such as magnesium stearate, calcium stearate, sodium stearyl fumarate or polyethylene glycol wax; sweetening agent; air freshener; syrup; etc. can be used. In addition, in the case of capsules, a liquid carrier such as fatty oil may be additionally used in addition to the above-mentioned materials.

A pharmaceutical composition for the prevention or treatment of IFTA containing the compound represented by Formula 1, its optical isomer or its pharmaceutically acceptable salt as an active ingredient may be formulated and used in various formulations suitable for parenteral administration.

Preparations for parenteral administration include, but are not limited to, injection solutions, suppositories, powders for respiratory inhalation, aerosols for sprays, ointments, powders for application, oils, creams, and the like.

In addition, in order to formulate the pharmaceutical composition according to the present invention as a preparation for parenteral administration, sterilized aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried preparations, external preparations, and the like may be used. Specifically, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate may be used as non-aqueous solvents and suspensions.

When the pharmaceutical composition according to the present invention is formulated as an injection solution, the pharmaceutical composition is mixed in water with a stabilizer or buffer to prepare a solution or suspension, which is formulated for unit administration in an ampoule or vial. can In addition, when the pharmaceutical composition according to the present invention is formulated as an aerosol, a propellant or the like may be blended with additives so that the water-dispersed concentrate or wet powder is dispersed. In addition, when the pharmaceutical composition according to the present invention is formulated into an ointment, cream, etc., animal oil, vegetable oil, wax, paraffin, starch, tracanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, oxidation It can be formulated using zinc or the like as a carrier.

A pharmaceutical composition according to the present invention may further comprise one or more other therapeutic agents suitable for IFTA treatment.

One or more other therapeutic agents suitable for IFTA treatment may be selected from the group consisting of orthoticosteroid drugs, non-steroidal anti-inflammatory drugs (NSAIDs), immunosuppressive agents, cytotoxic drugs and vasodilators. More specifically, ACE inhibitors, ARBs or statins, corticosteroids or calcinerone inhibitors (eg, tacrolimus), but not limited thereto. In addition, one or more other therapeutic agents suitable for IFTA treatment may be administered together or separately. When administered separately, they may be administered simultaneously or sequentially in any order. The dosage and timing of administration of the compound of Formula 1 or a pharmaceutically acceptable salt thereof and other therapeutic agents can be selected to achieve the desired combined therapeutic effect.

The pharmaceutical composition according to the present invention may further include a pharmaceutically acceptable carrier. "Pharmaceutically acceptable" means that which is commonly used in the field of pharmacology, which does not stimulate the organism upon administration of the compound and does not interfere with the biological activity and properties of the administered compound.

The type of carrier is not particularly limited, and any carrier commonly used in the art may be used. Non-limiting examples of the carrier include saline, sterile water, Ringer's solution, buffered saline, albumin injection solution, lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, maltodextrin, glycerol, ethanol, and the like. there is. These may be used alone or in combination of two or more.

In addition, if necessary, other pharmaceutically acceptable additives such as excipients, diluents, antioxidants, buffers or bacteriostats may be added to the pharmaceutical composition according to the present invention. In addition, if necessary, fillers, extenders, wetting agents, disintegrants, dispersants, surfactants, binders or lubricants may be additionally added to the pharmaceutical composition according to the present invention.

The pharmacologically effective amount of the pharmaceutical composition according to the present invention and the dosage to the human body may vary depending on the formulation method, administration method, administration time and/or administration route of the pharmaceutical composition. The above dose depends on the type and degree of response to be achieved by administration of the pharmaceutical composition, the type of subject to be administered, age, weight, general health condition, symptom or degree of disease, sex, diet, excretion, subject It may vary according to various factors including components of other compositions such as drugs used simultaneously or at this time and similar factors widely known in the medical field, and those skilled in the art can effectively administer the desired treatment The dosage can be easily determined and prescribed. For example, the human dose of the pharmaceutical composition according to the present invention may be 0.1 mg/kg to 1,000 mg/kg, preferably 10 mg/kg to 500 mg/kg per day. In addition, administration of the pharmaceutical composition according to the present invention may be administered once a day, or divided into several administrations. In addition, the administration route and administration method of the pharmaceutical composition according to the present invention may be independent, respectively, and the method is not particularly limited. Any administration route and administration method may be followed as long as the pharmaceutical composition according to the present invention is administered and the active ingredient can reach the target site.

The pharmaceutical composition according to the present invention may be administered by oral administration or parenteral administration. For example, as a parenteral administration method, intravenous administration, intraperitoneal administration, intramuscular administration, transdermal administration, or subcutaneous administration may be used. In addition, a method of applying, spraying, or inhaling the pharmaceutical composition according to the present invention to the diseased area may be used, but is not limited thereto. In addition, the pharmaceutical composition according to the present invention can be used for IFTA treatment in a subject in need thereof. In this case, the type of subject is not particularly limited, but may be a mammal, preferably a human.

The S1PR1 receptor is mainly present in lymphocytes, and is mainly responsible for regulating the release of lymphocytes from immune cells. The S1PR4 receptor is also present in lymphocytes and acts not only on immune cell trafficking, but also on T cell regulation, and is involved in inflammatory responses. Therefore, the compound of the present invention acts as a functional inhibitor to the S1PR1 receptor, inhibits the release of lymphocytes, and acts on S1PR4 and exhibits an effective anti-inflammatory effect, thereby preventing or treating IFTA.

Specifically, the compound of the present invention has the effect of preventing or treating IFTA by acting as a functional antagonist for these receptors through a mechanism that specifically binds to S1PR1 and S1PR4 and then enters and disappears them into cells. In addition, the pharmaceutical composition according to the present invention acts as a functional inhibitor for S1PR1 and S1PR4 and may not cause cardiovascular disease side effects.

Hereinafter, the present invention will be described in more detail through examples. These examples are only for exemplifying the present invention, and it will be apparent to those skilled in the art that the scope of the present invention is not to be construed as being limited by these examples.

Example - Preparation of Compound Represented by Formula 1

The compound represented by Formula 1 of the present invention can be prepared through a method as described in Korean Patent Publication No. 10-2017-0087813, and the method is the same as in the examples described below. However, the preparation method of the compound represented by Chemical Formula 1 of the present invention is not limited thereto, and may be prepared by a modified method within a range changeable by a person skilled in the art.

Example 1 - Preparation of 2-amino-2-(2-(1-decyl-1H-1,2,3-triazol-4-yl)ethyl)propane-1,3-diol

Step 1: Preparation of tert-butyl(2,2-dimethyl-5-((triisopropylsilyl)buta-1,3-diyn-1-yl)-1,3-dioxan-5-yl)carbamate manufacturing

To a solution of methanol (50 mL) in which CuCl (10 mg, 0.10 mmol), NH2OH HCl (1.02 g, 14.67 mmol), and n-BuNH ₂ (3.87 mL, 39.15 mmol) were dissolved under N ₂ gas conditions, methanol ( 30 mL) dissolved in tert-butyl 5-ethynyl-2,2-dimethyl-1,3-dioxan-5-yl carbamate (1.25 g, 4.89 mmol) was added, and then dissolved in methanol (20 mL). 2-Bromo-1-triisopropylsilylacetylene (2.56 g, 9.78 mmol) was added dropwise and the reaction mixture was stirred at room temperature for 2 hours, then the reaction was quenched with water and concentrated. After diluting this concentrate with ethyl acetate and washing with brine, the organic layer was dried over MgSO ₄ , filtered in vacuo and concentrated. This was purified by flash column chromatography (hexane/EtOAc, 7:1) to obtain the target compound as a white solid (1.85 g, 4.25 mmol, 87 %).

Step 2: tert-Butyl(5-((1-decyl-1H-1,2,3-triazol-4-yl)ethynyl)-2,2-dimethyl-1,3-dioxan-5-yl ) Preparation of carbamates

In an anhydrous DMF (12 mL) solution in which the compound (334 mg, 1.20 mmol) prepared in step 1 was dissolved, 1-azidodecane (339 mg, 1.86 mmol), CuI (114 mg, 0.60 mmol), DIPEA (N , N-Diisopropylethylamine) (0.63 mL, 3.6 mmol) and AgF (182 mg, 1.44 mmol) were added. After stirring the reaction mixture well at room temperature for 12 hours, the reaction was quenched with saturated NH ₄ Cl, extracted twice with ethyl acetate, and the organic layer extract was washed with saturated NH ₄ Cl and brine. It was dried over MgSO ₄ and concentrated under reduced pressure. The concentrate was purified by flash column chromatography (hexane/EtOAc, 6:1) to obtain the target compound as a white solid (439 mg, 0.95 mmol, 79%).

Step 3: tert-Butyl(5-(2-(1-decyl-1H-1,2,3-triazol-4-yl)ethyl)-2,2-dimethyl-1,3-dioxane-5- 1) carbamate

To a solution of the compound (149 mg, 0.32 mmol) prepared in step 2 in methanol (30 mL), 10% Pd/C (45 mg, 30% wt of the compound prepared in step 2) was added. After vacuuming the flask, filling it with H ₂ gas, stirring at room temperature for 1 hour, filtering the mixture through Celite, concentrating, and precipitate using flash column chromatography (hexane/EtOAc, 7:1 or 2: 1) to give the target compound as a colorless oil (110 mg, 0.24 mmol, 74%).

Step 4: Preparation of 2-amino-2-(2-(1-decyl-1H-1,2,3-triazol-4-yl)ethyl)propane-1,3-diol

TFA (1.40 mL) was added to a solution of CH ₂ Cl ₂ (1.40 mL) in which the compound prepared in step 3 (65 mg, 0.14 mmol) was dissolved at room temperature. After stirring the reaction for 12 hours and removing the solvent and reagent under reduced pressure, the concentrate was purified by flash column chromatography (CH ₂ Cl ₂ /MeOH/NH ₄ OH, 100:10:1) to obtain the target compound as a white solid. prepared (36 mg, 0.11 mmol, 78%).

¹ H NMR (400 MHz, CDCl ₃ ) δ 0.86 (t, J = 6.8 Hz, 3H), 1.23-1.29 (m, 14H), 1.85 (t, J = 6.8 Hz, 2H), 1.92 (t, J = 7.7 Hz, 2H), 2.47 (br s, 4H), 2.78 (t, J = 7.8 Hz, 2H), 3.57 (dd, J = 11.2, 26.9 Hz, 4H), 4.27 (t, J = 7.3 Hz, 2H) ), 7.31 (s, 1H); ¹³ C NMR (75 MHz, CDCl ₃ ) δ 14.03, 19.21, 22.57, 26.50 (2C), 28.95 (2C), 29.03 (2C), 30.19, 31.70, 50.42, 60.05, 63.09 (2C), 121.34, 146.82; IR (CHCl ₃ ) υmax; HRMS (FAB) calcd for C ₁₇ H ₃₅ N ₄ O ₂ ([M+H] ⁺ ) 327.2760, found 327.2762.

Example 2 - Preparation of 2-amino-2-(2-(1-octyl-1H-1,2,3-triazol-4-yl)ethyl)propane-1,3-diol

Instead of using 1-azidodecane, the target compound was prepared in the same manner as in Example 1, except that 1-azidooctane was used.

¹ H NMR (400 MHz, CDCl ₃ ) δ 0.86 (t, J = 6.8 Hz, 3H), 1.24-1.29 (m, 10H), 1.84-1.91 (m, 4H), 2.48 (br s, 4H), 2.77 (t, J = 7.4 Hz, 2H), 3.54 (q, J = 12.2 Hz, 4H), 4.28 (t, J = 7.2 Hz, 2H), 7.28 (s, 1H).

Example 3 - 2-amino-2-(2-(1-(4-hexylphenethyl)-1H-1,2,3-triazol-4-yl)ethyl)propane-1,3-diol manufacturing

The target compound was prepared in the same manner as in Example 1, except that 1-(2-azidoethyl)-4-hexylbenzene was used instead of 1-azidodecane.

Example 4 - Preparation of 2-amino-2-(1-dodecyl-1H-1,2,3-triazol-4-yl)propane-1,3-diol

Bromotriisopropylsilane was used instead of 2-bromo-1-triisopropylsilyl acetylene used in step 1 of Example 1, and 1-aza instead of 1-azidodecane in step 2 A target compound was prepared in the same manner as in Example 1, except that 1-azidodoedcane was used.

^1H NMR (300 MHz, MeOD) δ 0.89 (t, J = 6.7 Hz, 3H), 1.28-1.32 (m, 18H), 1.90-1.92 (m, 2H), 3.89-3.98 (m, 4H), 4.41 (t, J = 7.1 Hz, 2H), 8.07 (s, 1H).

Example 5 - Preparation of (E)-2-amino-2-(1-decyl-1H-1,2,3-triazol-4-yl)vinyl-1,3-diol

After the temperature of the THF (5 mL) solution in which the compound prepared in step 2 of Example 1 was dissolved was lowered to -78 ° C, a THF solution (0.3 mmol) in which LAH (lithium aluminum hydride) was dissolved was slowly added dropwise. After stirring at 0 °C for 3 hours, the reaction was quenched with water and extracted twice with CH ₂ Cl ₂ . The organic layer was washed with brine, dried over MgSO ₄ and concentrated in vacuo. The concentrate was purified by flash column chromatography (CH ₂ Cl ₂ /MeOH, 20:1) to obtain the target compound as a white solid. Thereafter, the target compound was prepared in the same manner as in step 4 of Example 1. (24 mg, 0.074 mmol, 74%)

Example 6 - 2-amino-2-(2-(1-(8-phenyloctyl)-1H-1,2,3-triazolebutyl-4-yl)ethyl)propane-1,3-diol manufacturing

A target compound was prepared in the same manner as in Example 1, except that 8-azidooctylbenzene was used instead of 1-azidodecane used in step 2 of Example 1.

Example 7 - Preparation of N-(2-(1-dodecyl-1H-1,2,3-triazol-4-yl)-1,3-dihydroxypropan-2-yl)acetamide

Acetic anhydride (0.2 mL), MeOH (1 mL), and saturated NaHCO ₃ aqueous solution (1 mL) were added to the compound (50 mg, 1 eq, 0.15 mmol) prepared in Example 4, and stirred for 30 minutes, After concentrating and quenching the reaction with an aqueous ammonium chloride solution, extraction was performed twice with CH ₂ Cl ₂ , dried with MgSO ₄ , and concentrated to obtain the target compound in a yield of 98% without additional purification (54 mg, 98%). .

^1H NMR (300 MHz, MeOD) δ 0.89 (t, J = 6.7 Hz, 3H), 1.28-1.32 (m, 18H), 1.86-1.96 (m, 2H), 2.00 (s, 3H), 4.02 (q , J = 11.2 Hz, 4H), 4.35 (t, J = 7.2 Hz, 2H), 7.84 (s, 1H).

Example 8 - N-(4-(1-decyl-1H-1,2,3-triazol-4-yl)-1-hydroxy-2-(hydroxymethyl)butan-2-yl)acetamide manufacture of

A target compound was prepared by using the compound prepared in Example 1 as a starting material in a similar manner to the acetylation reaction performed in Example 7.

^1H NMR (300 MHz, MeOD) δ 0.86 (t, J = 6.8 Hz, 3H), 1.23-1.32 (m, 14H), 1.86-1.92 (m, 2H), 1.94 (s, 3H), 1.99-2.02 (m, 2H), 2.47 (m, 2H), 3.62 (dd, J = 11.2, 26.9 Hz, 4H), 4.31 (t, J = 7.3 Hz, 2H), 7.31 (s, 1H).

Example 9 - N-(4-(1-(4-hexylphenethyl)-1H-1,2,3-triazol-4-yl)-1-hydroxy-2-(hydroxymethyl)butane- 2-day) Preparation of acetamide

Using the compound prepared in Example 3 as a starting material, the target compound was prepared by carrying out similarly to the acetylation reaction performed in Example 7.

Experimental Example 1: Confirmation of inhibitory effect of NXC736 on mouse immune cells

FTY720 (Fingolimod) sequesters lymphocytes in secondary lymphoid organs by reversibly trapping some lymphocytes in the lymph node or bone marrow to inhibit their entry into the central nervous system or reduce the number of lymphocytes circulating in the bloodstream and activate them to reach the brain It is a multiple sclerosis treatment that has a mechanism of action that reduces the number of lymphocytes that are affected.

In order to confirm whether NXC736 acting on the S1P receptor also has an effect on reducing lymphocytes, the effect of NXC736 on immune cells was evaluated. After oral administration of 3 mg/kg/day NXC736 to mice, changes in lymphocytes in peripheral blood (PB) and bone marrow (BM) were classified into T cells, CD4 T cells, and CD8 T cells, and the test substance FACS analysis was performed at 0, 4, 8, 12, 24, and 36 hours after administration. The cell number was measured and expressed as a percentage (% of control) based on 0 time.

Referring to FIG. 1, by administering 3 mg/kg of NXC736, peripheral blood flow (PB) decreased to 40.6%, 16.4%, and 5.9% levels of T cells, CD4 T cells, and CD8 T cells after 0 hours after 4 hours. I was able to confirm. The decrease was maintained until 12 hours, then recovered and recovered to 88.9% at 36 hours.

In the bone marrow (BM), the distribution of T cells was confirmed at 135% after 4 hours at 0 hours and 131% after 24 hours after administration of 3 mg/kg of NXC736. This is believed to be a phenomenon that occurs when receptor internalization occurs by NXC736 and T cell release from the bone marrow (BM) to the peripheral bloodstream is suppressed.

After administering 3 mg/kg of NXC736, the cell number in T cells decreased from 4 hours, and it was confirmed that the inhibitory effect was maintained to some extent until 24 hours and then recovered. In conclusion, it was confirmed that administration of NXC736 reduced the number of lymphocytes circulating in the bloodstream and exhibited an immunosuppressive effect.

Experimental Example 2: Confirmation of the inhibitory effect of NXC736 on rat immune cells

To evaluate the effect of NXC736 on rat immune cells, 10 mg/kg/day of NXC736 was orally administered to SD rats, and then classified into T cells, CD4 T cells, and CD8 T cells in peripheral bloodstream (PB) and bone marrow (BM). and FACS analysis was performed at 0, 4, 8, 12, 24, 36, and 48 hours after administration of the test substance. The cell number was measured and expressed as a percentage (% of control) based on 0 time, and the number of T cells, CD4 T cells, and CD8 T cells was measured and expressed as % of control based on 0 time.

Referring to FIG. 2 , it was confirmed that T cells were reduced from 4 hours in peripheral blood flow (PB) after administration of NXC736. In the bone marrow (BM), on the contrary, there was no change or a tendency to increase.

Through this study, it was confirmed that NXC736 showed an immunosuppressive effect by reducing the number of lymphocytes in the peripheral bloodstream (PB) after oral administration. In addition, it was confirmed that there was no change or a tendency to increase in bone marrow (BM). These effects are thought to be related to the mechanism by which NXC736 inhibits lymphocyte liberation by acting as a functional antagonist on S1P receptors present in lymph nodes and bone marrow (BM).

Experimental Example 3: Confirmation of fibrosis inhibitory effect using mouse kidney fibrosis model

The antifibrotic efficacy of NXC736 in renal disease was evaluated using a unilateral urinary obstruction (UUO) model, which is a renal fibrotic disease model. After renal fibrosis was induced by ligation of one urinary tract of the mouse, 10, 30, 90 mg/kg/day of NXC736 and 30 mg/kg/day of Telmisartan, a positive control, were orally administered once a day for 2 weeks.

Referring to FIG. 3, it was confirmed that renal fibrosis was induced as the fibrosis area (Sirius red positive area (%)) and kidney hydroxyproline in the vehicle were significantly increased compared to the negative control group (sham). Telmisartan, used as a positive control, decreased renal hydroxyproline statistically significantly (p<0.0001). Renal hydroxyproline was statistically significantly reduced (p<0.05, p<0.01, P<0.001) in all groups administered with 10, 30, and 90 mg/kg/day of NXC736, especially NXC736 90 mg/kg/day. It was confirmed that renal hydroxyproline was statistically significantly (p<0.05) reduced. As a result, it was confirmed that NXC736 was a substance with an inhibitory effect on fibrosis, and it was confirmed that NXC736 exhibited an inhibitory effect on renal fibrosis in a dose-dependent manner in a mouse kidney fibrosis model induced by UUO.

Experimental Example 4: Evaluation of the effect of NXC736 on the cardiovascular system

1) Evaluation of the effect of NXC736 on HEK293 cells in which the hERG gene is overexpressed

hERG (human Ether- -go-go Related Gene) was introduced and NXC736 was applied to HEK-293 cells stably expressing hERG potassium ion channels, and the effect on hERG channel currents was evaluated. NXC736 was set at concentrations of 1, 3, 10 and 30 μM. In addition, on the day of each experiment, one cell was treated with a negative control substance or one cell for which the recording of hERG channel currents for each concentration of test substance was completed.

Referring to FIG. 4, the compensated suppression rates (%) of hERG channel currents of NCX736 (groups B to E) at concentrations of 1, 3, 10, and 30 μM were 9.94 ± 1.96, respectively. 13.98±4.48, 35.03±12.09 and 82.62±7.31% (n=3), and the compensated suppression rate (%) of hERG channel currents in the negative control group (A group) was 0±4.12 ( n=3) values are shown. That is, it was confirmed that NCX736 (D and E groups) at 10 and 30 μM concentrations showed a statistically significant difference compared to the negative control group (A group).

E-4031 at a concentration of 0.1 μM was treated as a positive control group (F group) under the same conditions. The compensated suppression rate (%) of hERG channel currents showed a high value of 92.64±1.66% (n=5). In view of these results, this test method is judged to be an appropriate method to evaluate the effect of NCX736 on hERG channel currents.

In the evaluation of the effect on hERG channel currents using HEK-293 cells into which the hERG gene was introduced under the above conditions, the test substance NXC736 was treated at a concentration of up to 30 μM, resulting in a decrease in hERG channel currents. The compensated suppression rate (%) was 82.62 ± 7.31%, and the K ⁺ ion channel inhibition (IC50) was calculated as 12.94 μM (Hill coefficient: 1.527). Accordingly, it was confirmed that NXC736 does not inhibit hERG potassium channel activity and does not cause cardiac abnormalities.

2) Confirmation of effects on the cardiovascular system using beagle dogs

NXC736 was orally administered to 4 unanesthetized and unrestrained male beagle dogs with a remote transmitter inserted. Thereafter, the heart rate of the four beagle dogs was measured to evaluate the effect of NXC736 on the cardiovascular system, and the presence of abnormal symptoms was visually confirmed.

In this experiment, the test substance was administered to each of the four beagle dogs in four stages. In the first administration step (NXC736 0 mg/kg), 0.5% MC aqueous solution as a control substance was administered as an excipient, in the second administration step, NXC736 12.5 mg/kg was administered, and in the third administration step, NXC736 25 mg/kg was administered. In the fourth administration step, 50 mg/kg of NXC736 was administered. The 1st to 4th administration was performed at 1-week intervals. In addition, 0 hours (hours) before test substance administration, 0.5 hours (hours), 1 hour (hours), 2 hours (hours), 3 hours (hours), 4 hours (hours), 6 hours (hours) after test substance administration , After 8 hours (hours), 12 hours (hours), and 24 hours (hours), the heart rate of the beagle dogs was measured.

Referring to FIG. 5, it was confirmed that no change in heart rate was observed even when NXC736 was administered at doses of 12.5, 25, and 50 mg/kg.

In addition, as a result of observing general symptoms for the beagle dog with the naked eye, no abnormal symptoms were observed by administering NXC736 at doses of 12.5, 25, and 50 mg/kg.

As described above, the pharmaceutical composition comprising the compound represented by Formula 1 according to the present invention, its optical isomer or its pharmaceutically acceptable salt as an active ingredient acts as a functional inhibitor for S1PR1 and S1PR4 to prevent or treat IFTA. I was able to confirm that it worked. In addition, the pharmaceutical composition according to the present invention acts as a functional inhibitor for S1PR1 and S1PR4 among subtypes of S1P receptors (S1P1, S1P2, S1P3, S1P4 and S1P5), thereby preventing or treating IFTA and at the same time reducing cardiovascular disease side effects. It was confirmed that there was an effect that did not cause.

Claims (10)

A pharmaceutical composition for preventing or treating interstitial fibrosis and tubular atrophy (IFTA) comprising a compound represented by Formula 1, an optical isomer thereof or a pharmaceutically acceptable salt thereof as an active ingredient,
[Formula 1]


here,
R ¹ is hydrogen;
R ² is hydrogen or an acetyl group;
X is a single bond, C ₂ alkylene, or C ₂ alkenylene;
A is a 5-membered ring heteroarylene containing 3 N atoms;
B is C _2-11 straight or branched chain alkylene;
C is a single bond or phenylene; and
D is selected from the group consisting of hydrogen, phenyl and C _1-6 alkyl;
A pharmaceutical composition for preventing or treating interstitial fibrosis and tubular atrophy, characterized in that the compound represented by Formula 1 is any one compound selected from the group of compounds below:
(1) 2-amino-2-(2-(1-decyl-1H-1,2,3-triazol-4-yl)ethyl)propane-1,3-diol;
(2) 2-amino-2-(2-(1-octyl-1H-1,2,3-triazol-4-yl)ethyl)propane-1,3-diol;
(3) 2-amino-2-(2-(1-(4-hexylphenethyl)-1H-1,2,3-triazol-4-yl)ethyl)propane-1,3-diol;
(4) 2-amino-2-(1-dodecyl-1H-1,2,3-triazol-4-yl)propane-1,3-diol;
(5) (E)-2-amino-2-(1-decyl-1H-1,2,3-triazol-4-yl)vinyl-1,3-diol;
(6) 2-amino-2-(2-(1-(8-phenyloctyl)-1H-1,2,3-triazolebutyl-4-yl)ethyl)propane-1,3-diol;
(7) N-(2-(1-dodecyl-1H-1,2,3-triazol-4-yl)-1,3-dihydrooxypropan-2-yl)acetamide;
(8) N-(4-(1-decyl-1H-1,2,3-triazol-4-yl)-1-hydroxy-2-(hydroxymethyl)butan-2-yl)acetamide; and
(9) N-(4-(1-(4-hexylphenethyl)-1H-1,2,3-triazol-4-yl)-1-hydroxy-2-(hydroxymethyl)butane-2 -yl) acetamide.
delete delete delete delete According to claim 1,
A pharmaceutical composition for preventing or treating interstitial fibrosis and tubular atrophy, characterized in that the compound represented by Formula 1 is a compound represented by Formula 2 below:
[Formula 2]

According to claim 1 or 6,
The pharmaceutical composition for preventing or treating interstitial fibrosis and tubular atrophy, characterized in that the pharmaceutical composition is a preparation for oral administration or a preparation for parenteral administration.
According to claim 1 or 6,
The pharmaceutical composition for preventing or treating interstitial fibrosis and tubular atrophy, characterized in that the pharmaceutical composition further comprises at least one other therapeutic agent suitable for the treatment of transplant rejection.
According to claim 1 or 6,
The pharmaceutical composition is a pharmaceutical composition for preventing or treating interstitial fibrosis and tubular atrophy, characterized in that acts as a functional inhibitor for S1PR1 and S1PR4.
According to claim 1 or 6,
The pharmaceutical composition is a pharmaceutical composition for preventing or treating interstitial fibrosis and coronary atrophy, characterized in that it does not cause cardiovascular disease side effects.