Pharmaceutical composition for treating cockayne or bloom syndrome containing decursin derivatives as active ingredient

Abstract

The present invention relates to a pharmaceutical composition for treating Cockayne or Bloom syndrome, the pharmaceutical composition containing decursin derivatives as active ingredients, or a cosmetic composition for preventing or alleviating wrinkles, the cosmetic composition containing decursin derivatives as active ingredients. The decursin derivatives of the present invention effectively inhibit p16-dependent senescence in cells derived from Cockayne syndrome and Bloom syndrome patients, and thus can be applied as a therapeutic agent for Cockayne syndrome and Bloom syndrome.

Description

Pharmaceutical composition for treating cocaine or bloom syndrome containing a dekercin derivative as an active ingredient

[0001] The present invention relates to a pharmaceutical composition for treating cocaine or bloom syndrome, comprising a decusin derivative as an active ingredient, or a cosmetic composition for preventing or improving wrinkles, comprising a decusin derivative as an active ingredient.

[0002] Cockayne syndrome is a form of dwarfism that causes growth delay, photosensitivity, and premature aging. Cockayne syndrome is caused by a defect in a gene involved in the normal repair of DNA damaged by ultraviolet rays. Specifically, the gene responsible for Cockayne syndrome type A is the ERCC8 gene, located on chromosome 5, and the gene responsible for Cockayne syndrome type B is the ERCC6 gene, located on chromosome 10 (10q11).

[0003] Bloom syndrome is a rare genetic disorder that causes symptoms such as short stature, photosensitivity, facial telangiectasia, increased susceptibility to infections due to immunodeficiency, and increased susceptibility to cancer. It is caused by mutations in the genes that produce enzymes involved in DNA replication and repair, which maintain genome stability.

[0004] Cockayne syndrome and Bloom syndrome are both diseases caused by DNA repair defects, more specifically, they cause overexpression and accumulation of Lamin A, which induces abnormally strong binding of Lamin A to p53, which weakens the binding between BMI-1 and Lamin A, promotes the degradation of BMI-1, increases the expression of p16, and causes aging.

[0005] Based on this, the inventors of the present invention developed a leading drug capable of suppressing p16-dependent aging phenomenon caused by overexpression and accumulation of Lamin A, thereby completing the present invention.

[0006] The purpose of the present invention is to provide a pharmaceutical composition for treating progeria, which comprises a compound represented by the following chemical formula 1 and a pharmaceutically acceptable salt thereof as an active ingredient.

[0007] [Chemical Formula 1]

[0008]

[0009] (In the above chemical formula 1, R is , , , , , or ) is characterized by

[0010] Another object of the present invention is to provide a cosmetic composition for preventing or improving wrinkles, which comprises a compound represented by the above chemical formula 1 and a cosmetically acceptable salt thereof as an active ingredient.

[0011] The technical problems to be solved by the invention are not limited to the technical problems mentioned above, and other technical problems not mentioned can be clearly understood by a person having ordinary skill in the art from the description of the present invention.

[0012] The present invention provides a pharmaceutical composition for treating progeria, comprising a compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

[0013] [Chemical Formula 1]

[0014]

[0015] (In the above chemical formula 1, R is , , , , , or ) is characterized by

[0016] In the present invention, the progeria is characterized in that it is at least one disease selected from the group consisting of Cockayne syndrome and Bloom syndrome.

[0017] In the present invention, the compound represented by the above chemical formula 1 or a pharmaceutically acceptable salt thereof is characterized in that it treats progeria by inhibiting p16.

[0018] In addition, the present invention provides a cosmetic composition for preventing or improving wrinkles, comprising a compound represented by the above chemical formula 1 or a cosmetically acceptable salt thereof as an active ingredient.

[0019] In addition, the present invention provides a compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof.

[0020] In addition, the present invention provides a compound represented by the following chemical formula 2 or a pharmaceutically acceptable salt thereof.

[0021] [Chemical Formula 2]

[0022]

[0023] (In the above chemical formula 2, R is , , , or ) is characterized by

[0024] The present invention can provide a pharmaceutical composition for treating progeria, which comprises a compound represented by the above chemical formula 1 and a pharmaceutically acceptable salt thereof as an active ingredient.

[0025] In addition, the present invention can provide a cosmetic composition for preventing or improving wrinkles, which comprises a compound represented by the above chemical formula 1 and a cosmetically acceptable salt thereof as an active ingredient.

[0026] The effects of the present invention are not limited to the effects mentioned above, and other effects not mentioned will be clearly understood by those skilled in the art from the description of the claims.

[0027] Figure 1 is a diagram of compound 1 of the present invention. 1 This is a graph showing the H NMR spectrum.

[0028] Figure 2 is a diagram of compound 1 of the present invention. 13 This is a graph showing the C NMR spectrum.

[0029] Figure 3 is a graph showing the HRMS spectrum of compound 1 of the present invention.

[0030] Figure 4 is a graph showing the results of 2D NMR analysis of compound 1 of the present invention.

[0031] Figure 5 is a graph showing the HMBC spectrum of compound 1 of the present invention.

[0032] Figure 6 is a graph showing the HSQC spectrum of compound 1 of the present invention.

[0033] Figure 7 is a graph showing the COSY spectrum of compound 1 of the present invention.

[0034] Figure 8 is a diagram of compound 2 of the present invention. 1 This is a graph showing the H NMR spectrum.

[0035] Figure 9 is a drawing showing the results of immunofluorescence staining showing changes in the expression of Lamin A / C, p16, and H3K9me3 by compound 1 of the present invention.

[0036] Figure 10 is a graph showing the nuclear deformation recovery effect of compound 1 of the present invention, and is a graph calculating the percentage of cells with nuclear deformation.

[0037] Figure 11 is a drawing analyzing the aging-related beta-galactosidase activity (SA-β-gal activity) of compound 1 of the present invention.

[0038] Figure 12 is a graph showing the number of β-gal positive cells calculated by treatment with compound 1 of the present invention.

[0039] Figure 13 is a graph showing the cell proliferation recovery effect by compound 1 of the present invention.

[0040] Figure 14 is a graph showing the results of analyzing changes in p16 promoter activity by compounds 1 to 6 of the present invention.

[0041] Figure 15 is a Western blot result showing changes in p16 protein expression by compounds 1 to 6 of the present invention.

[0042] Figure 16 is a schematic diagram showing the p16 inhibition mechanism of compounds 1 to 6 of the present invention.

[0043] The terms used in this specification have been selected from widely used, current terms, taking into account the functions of the present invention. However, these terms may vary depending on the intentions of those skilled in the art, precedents, the emergence of new technologies, etc. Furthermore, in certain cases, terms may be arbitrarily selected by the applicant, and in such cases, their meanings will be described in detail in the relevant description of the invention. Therefore, the terms used in this invention should not be defined simply as names, but rather based on their inherent meanings and the overall content of the present invention.

[0044] Unless otherwise defined, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. Terms defined in commonly used dictionaries should be interpreted as having a meaning consistent with their meaning in the context of the relevant technology, and shall not be interpreted in an idealized or overly formal sense unless explicitly defined herein.

[0045] Numerical ranges are inclusive of the numbers defined in the ranges above. Every maximum numerical limitation given throughout this specification includes every lower numerical limitation, as if that lower numerical limitation were explicitly stated. Every minimum numerical limitation given throughout this specification includes every higher numerical limitation, as if that higher numerical limitation were explicitly stated. Every numerical limitation given throughout this specification will include every better numerical range within that broader numerical range, as if that narrower numerical limitation were explicitly stated.

[0046] Throughout this specification, '%' used to indicate the concentration of a particular substance means solid / solid (w / w) %, solid / liquid (w / v) %, and liquid / liquid (v / v) %, unless otherwise stated.

[0047]

[0048] Hereinafter, the present invention will be described in detail.

[0049]

[0050] Deckersin derivatives

[0051] The present invention can provide a decusin derivative compound represented by the following chemical formula 1.

[0052] [Chemical Formula 1]

[0053]

[0054] In the above chemical formula 1, R is , , , , , or It may be characterized by, and preferably or It may be, and more preferably, It could be.

[0055]

[0056] Pharmaceutical composition for the treatment of progeria

[0057] The present invention can provide a pharmaceutical composition for treating an aging-related disease, comprising a compound represented by the above chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient, and preferably, a pharmaceutical composition for treating progeria, comprising a compound represented by the above chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

[0058] The above-mentioned age-related disease may be at least one disease selected from the group consisting of Cockayne syndrome, Bloom syndrome, progeria, Werner syndrome, Hutchinson-Gilford syndrome, xeroderma pigmentosum, Rotmund-Thomson syndrome, and Down syndrome, preferably at least one disease selected from the group consisting of Cockayne syndrome and Bloom syndrome, and more preferably Cockayne syndrome or Bloom syndrome.

[0059] The pharmaceutical composition according to the present invention may be formulated and used in the form of external preparations such as powders, granules, sustained-release granules, enteric-coated granules, liquids, eye drops, ellipsoids, emulsions, suspensions, alcohols, troches, aromatic waters, limonades, tablets, sustained-release tablets, enteric-coated tablets, sublingual tablets, hard capsules, soft capsules, sustained-release capsules, enteric capsules, pills, tinctures, soft extracts, dry extracts, fluid extracts, injections, capsules, irrigation solutions, ointments, lotions, pastes, sprays, inhalants, patches, sterile injection solutions, or aerosols, according to a conventional method, and the external preparations may have formulations such as creams, gels, patches, sprays, ointments, ointments, lotions, liniments, pastes, or cataplasmas.

[0060] The pharmaceutical composition of the present invention may further comprise one or more additives selected from the group consisting of suitable carriers, excipients, disintegrants, sweeteners, coating agents, swelling agents, lubricants, glidants, flavoring agents, antioxidants, buffers, bacteriostatic agents, diluents, dispersants, surfactants, binders, and lubricants commonly used in the manufacture of pharmaceutical compositions.

[0061] Specifically, carriers, excipients, and diluents may include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, and mineral oil. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like. These solid preparations may be prepared by mixing at least one excipient, for example, starch, calcium carbonate, sucrose or lactose, gelatin, and the like, into the composition. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Liquid preparations for oral administration include suspensions, solutions, emulsions, and syrups. In addition to commonly used simple diluents such as water and liquid paraffin, they may contain various excipients such as wetting agents, sweeteners, flavoring agents, and preservatives. Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solvents and suspending agents can be propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. Base materials for suppositories include witepsol, macrogol, Tween 61, cacao butter, laurin butter, and glycerogelatin.

[0062] The pharmaceutical composition of the present invention can be administered to mammals, including humans, by any method, and can be administered, for example, orally or parenterally. Parenteral administration methods include, but are not limited to, intravenous, intramuscular, intraarterial, intramedullary, intrathecal, intracardiac, transdermal, subcutaneous, intraperitoneal, intranasal, enteral, topical, sublingual, or rectal administration.

[0063] The term “pharmaceutically acceptable salt” used in the present invention refers to any organic or inorganic addition salt having a relatively non-toxic and harmless effective concentration for patients, and side effects caused by the salt do not reduce the atopic prevention, improvement or treatment effect of Reversine. For example, organic acids and inorganic acids or non-toxic salts can be used as free acids, and hydrochloric acid, phosphoric acid, sulfuric acid, nitric acid, tartaric acid, etc. can be used as inorganic acids, and methanesulfonic acid, p-toluenesulfonic acid, acetic acid, trifluoroacetic acid, maleic acid, succinic acid, oxalic acid, benzoic acid, tartaric acid, fumaric acid, mandelic acid, propionic acid, citric acid, lactic acid, glycolic acid, gluconic acid, galacturonic acid, glutamic acid, glutaric acid, Glucuronic acid, aspartic acid, ascorbic acid, carbonic acid, vanillic acid, hydroiodic acid, etc. can be used.

[0064]

[0065] Cosmetic composition for preventing or improving wrinkles

[0066] The present invention can provide a cosmetic composition for preventing or improving wrinkles, which comprises a compound represented by the above chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

[0067] The cosmetic composition of the present invention may be manufactured in the form of, but is not limited to, a toner, an essence, a cream, a pack, a gel, a lotion, an ointment, a patch, a foam, a spray, a soap, a bath additive, a spray, a nutritional lotion, a nutritional essence, a nutritional cream, a nutritional serum, a massage cream, a cosmetic bath additive, a body lotion, a body milk, a bath oil, a baby oil, a baby powder, a shower gel, a shower cream, a sunscreen lotion, a sunscreen cream, a suntan cream, a UV-blocking cosmetic, a cleansing milk, a skin whitening cream, a hand lotion, a hair lotion, a cosmetic cream, a bath soap, a liquid soap, a cosmetic soap, a hand cleanser, a cream soap, a facial wash, or a cosmetic soap.

[0068] The above cosmetic composition may further comprise suitable carriers, excipients, or diluents typically used in the manufacture of cosmetic compositions. Examples of such carriers, excipients, or diluents include, but are not limited to, purified water, oils, waxes, fatty acids, fatty alcohols, fatty acid esters, surfactants, absorbents, thickeners, antioxidants, viscosity stabilizers, chelating agents, buffers, lower alcohols, and the like. Furthermore, the composition may further comprise whitening agents, moisturizers, vitamins, sunscreens, perfumes, dyes, antibiotics, antibacterial agents, and antifungal agents, as needed.

[0069] The oil may include hydrogenated vegetable oil, castor oil, cottonseed oil, olive oil, palm oil, jojoba oil, avocado oil, and the wax may include beeswax, spermaceti, carnauba, candelilla, montan, ceresin, liquid paraffin, lanolin, and the like. The fatty acid may include stearic acid, linoleic acid, linolenic acid, oleic acid, and the like, the fatty acid alcohol may include cetyl alcohol, octyldodecanol, oleyl alcohol, panthenol, lanolin alcohol, stearyl alcohol, hexadecanol, and the like, and the fatty acid ester may include isopropyl myristate, isopropyl palmitate, butyl stearate, and the like. The surfactant may include cationic surfactants, anionic surfactants, and nonionic surfactants known in the art, and preferably, surfactants derived from natural products such as sodium cocoyl isethionate, sodium lauryl sulfoacetate, disodium laureth sulfosuccinate, disodium lauryl sulfosuccinate, decyl glucoside, decyl polyglucose, lauramidopropyl betaine, and coco betaine. In addition, it may include absorbents, thickeners, antioxidants, etc. widely known in the cosmetics field, and the types and amounts of these are as known in the art.

[0070] Hereinafter, examples of the present invention will be described in detail, but it is obvious that the present invention is not limited to the following examples.

[0071]

[0072] Example 1. Preparation of decusin derivatives

[0073] In Example 1, six types of decusin derivatives were prepared. The common preparation method for the six types of decusin derivatives is as shown in Scheme 1 below.

[0074]

[0075] [Reaction Formula 1]

[0076]

[0077]

[0078] In the above reaction scheme 1, 1 equivalent (eq) (314 mg, 1.28 mmol) of decurcinol was dissolved in dimethylformamide (7.5 mL), and 1.5 equivalents (eq) (286 mg, 1.92 mmol) of dichloropyrimidine was added to the mixture, and the mixture was stirred at 25°C for 30 minutes. 2 equivalents (eq) (831 mg, 2.55 mmol) of cesium carbonate (Cs2CO3) was added to the stirred mixture, and the mixture was stirred for 18 hours to obtain the target compound (421 mg, 91%).

[0079]

[0080] 1-1. Preparation of compound 1

[0081] In Example 1-1, compound 1 represented by the following structural formula 1 was prepared.

[0082] [Structural formula 1]

[0083]

[0084] The method for preparing the above compound 1 is as shown in the following reaction scheme 2.

[0085] [Reaction Formula 2]

[0086]

[0087] In the above reaction scheme 2, 1 equivalent (eq) (87 mg, 0.24 mmol) of the starting material was dissolved in 1,4-dioxane (2.4 mL). 2 equivalents (67 mg, 0.48 mmol) of 4-fluorobenzylboronic acid and 3 equivalents (76 mg, 0.72 mmol) of sodium carbonate (Na2CO3) were added to the mixture, and the mixture was stirred at 25°C for 1 minute. 0.2 equivalents (55 mg, 0.048 mmol) of tetrakis(triphenylphosphane)palladium was added to the stirred mixture, and the mixture was stirred at 80°C for 23 hours to obtain the target compound (71 mg, 71%).

[0088]

[0089] 1-2. Preparation of compound 2

[0090] In Example 1-2, compound 2 represented by the following structural formula 2 was prepared.

[0091] [Structural formula 2]

[0092]

[0093] The method for preparing the above compound 2 is as shown in the following reaction scheme 3.

[0094] [Reaction Formula 3]

[0095]

[0096] In the above reaction scheme 3, 1 equivalent (eq) (84 mg, 0.23 mmol) of the starting material was dissolved in 1,4-dioxane (2.4 mL), to which 2 equivalents (63 mg, 0.46 mmol) of 4-tolylboronic acid and 3 equivalents (73 mg, 0.69 mmol) of sodium carbonate (Na2CO3) were added, and the mixture was stirred at 25°C for 1 minute. 0.2 equivalents (53 mg, 0.046 mmol) of tetrakis(triphenylphosphane)palladium was added to the stirred mixture, and the mixture was stirred at 80°C for 17 hours to obtain the target compound (92 mg, 96%).

[0097]

[0098] 1-3. Preparation of compound 3

[0099] In Example 1-3, compound 3 represented by the following structural formula 3 was prepared.

[0100] [Structural formula 3]

[0101]

[0102] The method for preparing the above compound 3 is as shown in the following reaction scheme 4.

[0103] [Reaction Formula 4]

[0104]

[0105] In the above reaction scheme 4, 1 equivalent (eq) (84 mg, 0.23 mmol) of the starting material was dissolved in 1,4-dioxane (2.4 mL). 2 equivalents (77 mg, 0.46 mmol) of 4-methylthiobenzeneboronic acid and 3 equivalents (73 mg, 0.69 mmol) of sodium carbonate (Na2CO3) were added to the mixture, and the mixture was stirred at 25°C for 1 minute. 0.2 equivalents (53 mg, 0.046 mmol) of tetrakis(triphenylphosphane)palladium was added to the stirred mixture, and the mixture was stirred at 80°C for 2 hours to obtain the target compound (31 mg, 30%).

[0106]

[0107] 1-4. Preparation of compound 4

[0108] In Example 1-4, compound 4 represented by the following structural formula 4 was prepared.

[0109] [Structural formula 4]

[0110]

[0111] The method for preparing the above compound 4 is as shown in the following reaction scheme 5.

[0112] [Reaction Formula 5]

[0113]

[0114] In the above reaction scheme 5, 1 equivalent (eq) (84 mg, 0.23 mmol) of the starting material was dissolved in a mixture of 1,4-dioxane (2.4 mL), and 2 equivalents (eq) (76 mg, 0.46 mmol) of 3,4-(methylenedioxy)benzeneboronic acid and 3 equivalents (eq) (73 mg, 0.69 mmol) of sodium carbonate (Na2CO3) were added, and the mixture was stirred at 25°C for 1 minute. To the above stirred mixture, 0.2 equivalents (eq) (53 mg, 0.046 mmol) of tetrakis(triphenylphosphane)palladium was added, and the mixture was stirred at 80°C for 2 hours to obtain the target compound (90 mg, 87%).

[0115]

[0116] 1-5. Preparation of compound 5

[0117] In Example 1-5, compound 5 represented by the following structural formula 5 was prepared.

[0118] [Structural formula 5]

[0119]

[0120] The method for preparing the above compound 5 is as shown in the following reaction scheme 6.

[0121] [Reaction Formula 6]

[0122]

[0123] In the above reaction scheme 6, 1 equivalent (eq) (84 mg, 0.23 mmol) of the starting material was dissolved in 1,4-dioxane (2.4 mL). 2 equivalents (69 mg, 0.46 mmol) of 4-formylbenzeneboronic acid and 3 equivalents (73 mg, 0.69 mmol) of sodium carbonate (Na2CO3) were added to the mixture, and the mixture was stirred at 25°C for 1 minute. 0.2 equivalents (53 mg, 0.046 mmol) of tetrakis(triphenylphosphane)palladium was added to the stirred mixture, and the mixture was stirred at 80°C for 1.5 hours to obtain the target compound (78 mg, 80%).

[0124]

[0125] 1-6. Preparation of compound 6

[0126] In Example 1-6, compound 6 represented by the following structural formula 6 was prepared.

[0127] [Structural formula 6]

[0128]

[0129] The method for preparing the above compound 6 is as shown in the following reaction schemes 7 and 8.

[0130] [Reaction Formula 7]

[0131]

[0132] In the above reaction scheme 7, 1 equivalent (eq) (84 mg, 0.23 mmol) of the starting material was dissolved in 1,4-dioxane (2.4 mL). 2 equivalents (64 mg, 0.46 mmol) of 4-hydroxyphenylboronic acid and 3 equivalents (73 mg, 0.69 mmol) of sodium carbonate (Na2CO3) were added to the mixture, and the mixture was stirred at 25°C for 1 minute. 0.2 equivalents (53 mg, 0.046 mmol) of tetrakis(triphenylphosphane)palladium was added to the stirred mixture, and the mixture was stirred at 80°C for 2 hours to obtain the target compound (89 mg, 93%).

[0133]

[0134] [Reaction Formula 8]

[0135]

[0136] In the above reaction scheme 8, 1 equivalent (eq) (89 mg, 0.21 mmol) of the starting material was dissolved in tetrahydrofuran (6.0 mL), and 2 equivalents (eq) (52 mg, 0.43 mmol) of 4-dimethylaminopyridine and 1.3 equivalents (eq) (39 μl, 0.28 mmol) of triethylamine were added to the mixture, and then the temperature was lowered to 0°C. 1.5 equivalents (eq) (23 μl, 0.32 mmol) of acetyl chloride was added to the stirred mixture, and the mixture was stirred at 25°C for 10 minutes to obtain the target compound (93 mg, 95%).

[0137]

[0138] The structural formulas, chemical formulas, and molecular weights of compounds 1 to 6 prepared in Example 1 are listed in Table 1 below.

[0139] Compound 1 prepared in the above Example 1-1 1 H NMR spectrum is shown in Fig. 1. 13The C NMR spectrum is shown in Fig. 2, the HRMS spectrum is shown in Fig. 3, the 2D NMR analysis results are shown in Fig. 4, the HMBC spectrum is shown in Fig. 5, the HSQC spectrum is shown in Fig. 6, and the COSY spectrum is shown in Fig. 7. In addition, the compound 2 prepared in Example 1-2 is shown in Fig. 2. 1 The H NMR spectrum is shown in Figure 8.

[0140] NMR spectra were recorded on a Bruker AV VIII 400 or 600 spectrometer in the indicated solvents. Solvent signals were used as references, and chemical shifts were converted to the TMS scale (CDCl3: δC = 77.16 ppm; residual CHCl3 in CDCl3: δH = 7.26 ppm).

[0141] High-resolution mass spectrometry (HRMS) was measured using an ABSCIEXQ-TOF5600 mass spectrometer.

[0142]

[0143] Compound 1S)-7-((6-(4-fluorophenyl)pyrimidin-4-yl)oxy)-8,8-dimethyl-7,8-dihydro-2H,6H-pyrano[3,2-g]chromen-2-oneChemical Formula: C 24 H 19 FN2O4Molecular Weight: 418.4244 1H NMR (400 MHz, Chloroform-d) δ 8.81 (d,J= 1.0 Hz, 1H), 8.03 - 7.99 (m, 2H), 7.57 (d,J= 9.4 Hz, 1H), 7.18 - 7.13 (m, 3H), 7.06 (d,J= 1.1 Hz, 1H), 6.86 (s, 1H), 6.23 (d,J= 9.4 Hz, 1H), 5.53 (app. t,J= 4.7 Hz, 1H), 3.33 (dd,J= 17.3, 4.6 Hz, 1H), 3.04 (dd,J= 17.3, 4.7 Hz, 1H), 1.48 (s, 3H), 1.43 (s, 3H) 화합물 2(S)-8,8-dimethyl-7-((6-(p-tolyl)pyrimidin-4-yl)oxy)-7,8-dihydro-2H,6H-pyrano[3,2-g]chromen-2-oneChemical Formula: C 25 H 22 N2O4Molecular Weight: 414.4610 1 H NMR (400 MHz, Chloroform-d) δ 8.83 (d,J= 1.0 Hz, 1H), 7.93 - 7.89 (m, 2H), 7.57 (d,J= 9.4 Hz, 1H), 7.30 - 7.26 (m, 2H), 7.17 - 7.16 (m, 1H), 7.09 (d,J= 1.1 Hz, 1H), 6.86 (s, 1H), 6.23 (d,J= 9.5 Hz, 1H), 5.54 (app. t,J= 4.6 Hz, 1H), 3.33 (dd,J= 17.5, 4.6 Hz, 1H), 3.04 (dd,J= 17.5, 4.6 Hz, 1H), 2.41 (s, 3H), 1.49 (s, 3H), 1.43 (s, 3H) 화합물 3(S)-8,8-dimethyl-7-((6-(4-(methylthio)phenyl)pyrimidin-4-yl)oxy)-7,8-dihydro-2H,6H-pyrano[3,2-g]chromen-2-oneChemical Formula: C 25H 22 N2O4SMolecular Weight: 446.5210 1 H NMR (400 MHz, Chloroform-d) δ 8.89 (s, 1H), 8.00 - 7.97 (m, 2H), 7.57 (d,J= 9.4 Hz, 1H), 7.33 (s, 1H), 7.31 (s, 1H), 7.17 - 7.16 (m, 1H), 7.10 (d,J= 1.0 Hz, 1H), 6.86 (s, 1H), 6.24 (d,J= 9.4 Hz, 1H), 5.57 (app. t,J= 4.5 Hz, 1H), 3.34 (dd,J= 17.4, 4.5 Hz, 1H), 3.04 (dd,J= 17.5, 4.5 Hz, 1H), 2.53 (s, 3H), 1.49 (s, 3H), 1.44 (s, 3H) 화합물 4(S)-7-((6-(benzo[d][1,3]dioxol-5-yl)pyrimidin-4-yl)oxy)-8,8-dimethyl-7,8-dihydro-2H,6H-pyrano[3,2-g]chromen-2-oneChemical Formula: C 25 H 20 N2O6Molecular Weight: 444.4430 1 H NMR (400 MHz, Chloroform-d) δ 8.78 (d,J= 1.1 Hz, 1H), 7.58 - 7.57 (m, 1H), 7.56 - 7.55 (m, 1H), 7.51 - 7.52 (m, 1H), 7.17 - 7.15 (m, 1H), 6.99 (d,J= 1.0 Hz, 1H), 6.88 (d,J= 8.2 Hz, 1H), 6.85 (s, 1H), 6.23 (d,J= 9.5 Hz, 1H), 6.03 (s, 2H), 5.52 (app. t,J= 4.7 Hz, 1H), 3.32 (dd,J= 17.3, 4.7 Hz, 1H), 3.03 (dd,J= 17.2, 4.7 Hz, 1H), 1.48 (s, 3H), 1.43 (s, 3H) 화합물 5(S)-4-(6-((8,8-dimethyl-2-oxo-7,8-dihydro-2H,6H-pyrano[3,2-g]chromen-7-yl)oxy)pyrimidin-4-yl)benzaldehydeChemical Formula: C 25 H 20 N2O5Molecular Weight: 428.4440 1 H NMR (400 MHz, Chloroform-d) δ 10.09 (s, 1H), 8.88 (d,J= 1.0 Hz, 1H), 8.19 - 8.16 (m, 2H), 8.00 - 7.97 (m, 2H), 7.57 (d,J= 9.5 Hz, 1H), 7.19 (d,J= 1.1 Hz, 1H), 7.17 - 7.16 (m, 1H), 6.86 (s, 1H), 6.24 (d,J= 9.4 Hz, 1H), 5.56 (app. t,J= 4.7 Hz, 1H), 3.34 (dd,J= 17.3, 4.7 Hz, 1H), 3.05 (dd,J= 17.3, 4.6 Hz, 1H), 1.49 (s, 3H), 1.44 (s, 3H) 화합물 6(S)-4-(6-((8,8-dimethyl-2-oxo-7,8-dihydro-2H,6H-pyrano[3,2-g]chromen-7-yl)oxy)pyrimidin-4-yl)phenyl acetateChemical Formula: C 26 H 22 N2O6Molecular Weight: 458.4700 1H NMR (400 MHz, Chloroform-d) δ 8.82 (d,J= 1.0 Hz, 1H), 8.05 - 8.00 (m, 2H), 7.57 (d,J= 9.4 Hz, 1H), 7.22 - 7.21 (m, 1H), 7.20 - 7.19 (m, 1H), 7.17 - 7.16 (m, 1H), 7.08 (d,J= 1.1 Hz, 1H), 6.86 (s, 1H), 6.23 (d,J= 9.5 Hz, 1H), 5.53 (app. t,J= 4.7 Hz, 1H), 3.33 (dd,J= 17.4, 4.6 Hz, 1H), 3.04 (dd,J= 17.4, 4.7 Hz, 1H), 2.32 (s, 3H), 1.48 (s, 3H), 1.43 (s, 3H)

[0144]

[0145] Experimental Example 1. Therapeutic effect of compound 1 on cocaine and Bloom syndrome

[0146]

[0147] 1-1. Immunofluorescence staining

[0148] Cells derived from Cockayne syndrome patients (AG05012, AG06269) and Bloom syndrome patients (GM16886, GM16890) were obtained from Coriell Cell Repositories (New Jersey, USA) and cultured in EMEM medium containing 15% FBS, 26 mM antibiotic-free HEPES, and 2 mM glutamine. Nutlin-3 (N6287), used as a senescence stimulant, was purchased from Sigma.

[0149] Cells on cover glasses were washed with PBS, fixed with 4% PFA for 30 minutes at room temperature, and then permeabilized with 0.1% Triton X-100 / PBS for 10 minutes. A blocking solution was prepared by diluting anti-human antibody in PBS at a ratio of 1:500, and the cells were treated with the blocking solution for 1 hour, followed by reaction with anti-lamin A / C antibody, p16 antibody, and H3K9me3 antibody overnight at 4°C. At this time, anti-lamin A / C antibody, p16 antibody, and H3K9me3 antibody were used by diluting them with the blocking solution at a ratio of 1:500, 1:300, and 1:300, respectively. Finally, cells were incubated with FITC- and rhodamine-conjugated secondary antibodies at 4°C for 6 h, and nuclei were stained with 4,6-diamidino-2-phenylindole (DAPI) for 10 min. Cells were washed three times with PBS, and coverslips were mounted with mounting solution (H-5501, Vector Laboratories) and analyzed by fluorescence microscopy (Zeiss).

[0150] The results of the above Experimental Example 1-1 are shown in Fig. 9. Fig. 9A shows the results of immunofluorescence staining using cells derived from a cocaine patient, and Fig. 9B shows the results of immunofluorescence staining using cells derived from a bloom patient.

[0151] Referring to FIG. 9, p16 expression was reduced and H3K9me3 expression was increased in cells derived from patients with Cockayne or Bloom syndrome by compound 1 prepared in Example 1. Since high p16 expression and low H3K9me3 expression levels are representative examples of aging phenomena, it can be confirmed that the aging phenomenon was improved by compound 1 in cells derived from patients with Cockayne or Bloom syndrome.

[0152] Based on the results of Fig. 9, the percentage of cells with nuclear deformation following treatment with compound 1 was calculated and graphically represented in Fig. 10. Referring to Fig. 10, it was confirmed that the nuclear deformation phenomenon was recovered by reducing the number of cells with nuclear deformation following treatment with compound 1 in cells derived from Cockayne syndrome patients (AG05012, AG06269) and cells derived from Bloom syndrome patients (GM16886, GM16890).

[0153]

[0154] 1-2. Senescence-Associated-β-gal Activity Analysis

[0155] Cells derived from Cockayne syndrome patients (AG05152, GM00739) and cells derived from Bloom syndrome patients (GM16886) were cultured using the same method as in Experimental Example 1-1. For staining of senescence-associated beta-galactosidase activity (SA-β-gal activity), the cells were washed once with phosphate-buffered saline (PBS) (pH 7.2) and then fixed with PBS containing 0.5% glutaraldehyde. The cells were then washed with PBS and stained overnight at 37°C with X-gal solution (Cell Signaling Technology, Danvers, MA, USA). The results of Experimental Example 1-2 are shown in Fig. 11.

[0156] Referring to Figure 11, it can be confirmed that the aging phenomenon was improved by treating cells derived from Cockayne syndrome patients and Bloom syndrome patients with the compound 1, as shown by a decrease in aging-related beta-galactosidase activity.

[0157] Based on the results of Fig. 11, the percentage of beta-galactosidase positive cells according to compound 1 treatment was calculated and graphically represented in Fig. 12. Referring to Fig. 12, it was confirmed that beta-galactosidase positive cells were reduced by compound 1 treatment in cells derived from Cockayne syndrome patients (AG05012) and cells derived from Bloom syndrome patients (GM16886, GM16890).

[0158]

[0159] 1-3. Cell proliferation analysis

[0160] Cells derived from Cockayne syndrome patients (AG05152, GM00739) and cells derived from Bloom syndrome patients (GM16886) were cultured in the same manner as in Experimental Example 1-1 above. The cultured cells were cultured in 6-well plates at a low density (~5 × 10 5 Cells were seeded at 100 cells / well. After treatment with compound 1 prepared in Example 1-1, cells were collected and stained with Trypan blue (GIBCO, Grand Island, NY, USA) at room temperature for 5 minutes. The number of unstained cells (viable cells) was counted using a hemocytometer. The results of Experimental Example 1-3 are shown in Fig. 13.

[0161] Referring to Figure 13, the group treated with compound 1 in cells derived from Cockayne syndrome patients (AG05152, GM00739) and cells derived from Bloom syndrome patients (GM16886) showed a better cell proliferation effect than the control group, and it was confirmed that compound 1 had an effect of restoring cell proliferation.

[0162]

[0163] Experimental Example 2. p16 inhibitory effect of compounds 1 to 6

[0164]

[0165] 2-1. p16 promoter activity analysis (Luciferase assay)

[0166] To evaluate the transcriptional activity of p16, the HCT116 colon cancer cell line was used. The colon cancer cell line was provided by Dr. B. Vogelstein (Johns Hopkins University) and cultured in RPMI1640 medium containing 10% FBS and 1% antibiotics at 37°C and 5% CO2.

[0167] To evaluate the transcriptional activity of p16, the cultured cells were co-transfected with the p16 luciferase vector and the Lamin A vector for 24 hours. Thereafter, the cells were incubated with compounds 1 to 6 (10 μM) prepared in Example 1 for 24 hours. After washing with PBS, the cells were lysed with lysis buffer (Promega, Madison, WI, USA). Luciferase activity was measured using a luminometer (Micro-Digital, Gyeonggi-do, South Korea). Transfection was performed using Jet-PEI reagent (JetPEI; Polyplus transfection, New York, NY, USA) according to the manufacturer's protocol. The results of Experimental Example 2-1 are graphically represented in Figure 14.

[0168] Referring to FIG. 14, it can be confirmed that p16 transcriptional activity is reduced by compounds 1 to 6 prepared in Example 1, and among them, it can be confirmed that compound 1 has the best p16 transcriptional activity inhibitory effect.

[0169]

[0170] 2-2. Western blot analysis

[0171] To determine whether p16 protein expression was inhibited, the HEK293 renal cell line was used. The renal cell line was obtained from the American Type Culture Collection (USA) and cultured in DMEM medium containing 10% FBS and 1% antibiotics at 37°C and 5% CO2.

[0172] Proteins were extracted from the cultured cells using lysis buffer (50 mM Tris-Cl, pH 7.5, 150 mM NaCl, 1% NP-40, 0.1% SDS, and 10% sodium deoxycholate). The samples were separated by SDS-PAGE and transferred to PVDF membranes. The blotted membranes were blocked with 3% skim milk containing TBST buffer for 1 hour, incubated with antibodies, and then detected using ECL solution. The antibodies used in Experimental Example 2-2 are as follows: Actin antibody (sc-47778, Santa Cruz Biotechnology); BMI-1 antibody (5856, Cell Signaling Technology); p16 antibody (10883-1-AP, Proteintech); The results of Experimental Example 2-2 are shown in Figure 15.

[0173] Referring to FIG. 15, it can be confirmed that the protein expression of p16 was reduced by compounds 1 to 6 prepared in Example 1, and in particular, it was confirmed that the p16 inhibitory effect of compounds 1 and 6 was excellent.

[0174]

[0175] From the above description, those skilled in the art will understand that the present invention can be implemented in other specific forms without altering its technical spirit or essential features. In this regard, it should be understood that the embodiments described above are illustrative in all respects and are not limiting.

Claims

1. A pharmaceutical composition for treating progeria, comprising a compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient. [Chemical Formula 1] 2. In paragraph 1, R in the above chemical formula 1 is , , , , , or A pharmaceutical composition for treating progeria, characterized by:

3. In paragraph 1, The above symptoms are A pharmaceutical composition for treating progeria, characterized in that it is at least one disease selected from the group consisting of Cockayne syndrome and Bloom syndrome.

4. In paragraph 1, The compound represented by the above chemical formula 1 or a pharmaceutically acceptable salt thereof A pharmaceutical composition for treating progeria, characterized in that it treats progeria by inhibiting p16.

5. A cosmetic composition for preventing or improving wrinkles, comprising a compound represented by the following chemical formula 1 or a cosmetically acceptable salt thereof as an active ingredient. [Chemical Formula 1] 6. In paragraph 5, R in the above chemical formula 1 is , , , , , or A cosmetic composition for preventing or improving wrinkles, characterized by:

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