KR100520096B1 - Phenylamidine derivatives having an antioxidative activity and pharmaceutical compositions containing them - Google Patents

Abstract

The present invention relates to a phenylamidine derivative having excellent antioxidant activity and a pharmaceutical composition comprising the same. More specifically, the phenylamidine derivative represented by the following Formula 1 is not only excellent in antioxidant activity compared to conventional antioxidant compounds, Its low toxicity makes it useful for anti-aging, cancer, diabetes and epilepsy treatments, and for dementia, Parkinson's disease, stroke and neurodegenerative diseases of Huntington.

Formula 1

(Wherein X ₁ , X ₂ , Y and Ar are as described in the specification).

Description

Phenylamidine derivative having an antioxidant activity and a pharmaceutical composition comprising the same {PHENYLAMIDINE DERIVATIVES HAVING AN ANTIOXIDATIVE ACTIVITY AND PHARMACEUTICAL COMPOSITIONS CONTAINING THEM}

The present invention relates to phenylamidine derivatives and uses thereof, and more particularly, to phenylamidine derivatives and pharmaceutically acceptable salts thereof, which exhibit excellent antioxidant activity compared to conventional antioxidant compounds, and pharmaceutical compositions using the derivatives as active ingredients. It is about.

The oxygen serves to produce ATP by oxidation of nutrients in the mitochondria in the cells (mitochondria), but 2-5% of the oxygen is inevitably superoxide radicals ^{_{(superoxide radical, · O 2 -}} ) in the air we breathe, Hydrogen peroxide (H ₂ O ₂ ), hydroxy radicals (Hy radicals, etc.), such as reactive oxygen (Hydrogen radicals) harmful to the human body (Reactive Oxygen Species) will be changed.

As a defense against them, there are antioxidants such as superoxide dismutase (SOD), catalase, and peroxidase, and antioxidants such as glutathione and coenzyme Q (CoQ). They also eat antioxidants from food. However, in certain circumstances, such as ischemia, excess free radicals may occur, exceeding the capacity of the biological defense system, leading to oxidative stress. These free radicals attack biological cells, destroying lipids, proteins and nucleic acids (RNA, DNA) and inhibiting various enzyme functions, leading to diseases and aging, such as cancer, neurodegenerative diseases, and cardiovascular diseases. Results in.

Accordingly, antioxidants that eliminate or inhibit free radicals may be used as anti-aging or anticancer agents, as well as for treating various neurodegenerative diseases such as dementia and stroke.

Specifically, active oxygen is non-selective and irreversible destructive action on the cell components, such as lipid (Lipid), protein (Protein), sugar (Sugar), DNA (Deoxyribonucleic acid). Low density lipoproteins or antioxidants (Enzymes) naturally produced in plasma act to protect cells from oxidation. Prevention of lipid peroxidation in respiratory organisms is a fundamental action, and in organic respiratory organisms it protects reactive oxygen species ("ROS") caused by respiration and oxidation of substrates. It has a defense system. A small amount of _{ROS (· OH, · O 2} -, H 2 O 2), and further the nitric oxide radical (nitric oxide; NO ·), peroxide, nitrite (peroxynitrite; ONOO ^-) is breathing was stirred by the internal and external It is constantly being produced in the process. At low levels, ROS plays an indispensable role in biochemical processes such as defense against microorganisms, internal signaling systems in the immune system, cell differentiation and growth. On the contrary, if the amount of ROS exceeds the amount that can be processed in vivo, it may cause biological damage and oxidative stress such as dysfunction in the metabolism. Failure to remove this oxidative toxicity can damage DNA or interfere with the production of proteins such as Na ⁺ / K ⁺ ATPases and glutamate transporters. In addition, if lipid peroxidation increases and antioxidant action decreases, the nucleus center of the cell is attacked to damage proteins, DNA, and RNA. Allergies, mutations, or carcinogenesis may occur.

The oxidative stress of ROS that can cause this damage can be explained in relation to various pathological symptoms in the central nervous system (CNS), mainly physical processes such as neurodegenerative diseases and aging. have. The reason why ROS is particularly fatal to nervous system disease is that the nervous system has very fast oxygen metabolism, low level of antioxidant defense against oxidative action, and high content of unsaturated fatty acid and transition metal, which is significant for oxidative stress. be sensitive. Thus, if the production of ROS increases and fails to eradicate it, the micromolecules of the cell are significantly damaged. As a result, DNA, proteins, and lipids are damaged, resulting in structural and functional abnormalities. In particular, brain lipid damage caused by peroxidation of lipids causes severe neurological diseases (Neurodegenerative Diseases).

When divided into acute and chronic diseases, acute neurological diseases include ischemic stroke, subarachnoid hemorrhage, trauma to the Brain and Spinal cord. Neurological diseases include Alzheimer's disease, Parkinson's disease, and Huntington's disease. Among them, the mortality rate due to stroke, which is representative of cerebrovascular disease, is increasing every year in Korea as well as in the world.

The field of research on the development of therapeutic agents for stroke is the field of glutamate receptor antagonists, among which N- methyl- D- aspartate (NMDA) receptor antihormones and AMPA ( α- amino-3- hydroxy-4-isoxazole propionic acid receptor anti-hormone, γ-aminobutyric acid (GABA) receptor anti-hormone, calcium channel blocker, sodium channel blocker, etc. There is a great deal of interest in the development of treatments for stroke [Barr, PR; Flint Beal, M. Neuroprotection in CNS Diseases ; Marcel Dekker Inc.].

For many years, antioxidants that eliminate human harmful substances have been successfully used to protect synthetics, foods, and the like from oxidation. Recently, researches and developments have been conducted on neuroprotective drugs for various diseases caused by oxidative stress (Parkinson's Disease, Alzheimer's Diseases, stroke, traumatic injury). Antioxidants, as mentioned above, play a role in maintaining the level of free radicals in our body that are produced during the process caused by respiratory action in our body. AOEs) and endogenous organic compounds; reduced glutathione or GSH, ubiquinone or coenzyme Q, NADPH, melatonin, uric acid, etc., trace minerals (Se, Zn, Mn), vitamins (vitamin A, The ROS free radicals are treated by an interoperative complex system of antioxidants such as C, and E). However, if an excess amount of ROS is generated beyond this system and causes a disease state, chemically synthesized antioxidants are used to solve this problem. Antioxidants of tert- butyl-hydroxytoluene (BHT), idebenone, carbazole and phenazine series have already been developed for such synthesized antioxidants [Okamoto, K .; Wasazumi, M .; Morimoto, H .; Imada, I. Chem. Pharm. Bull. 1988, 36, 178. Yamaguchi, T .; Sano, K .; Takakura, K .; Saito, I .; Shinohara, Y .; Asano, T .; Yasuhara, H. Stroke 1998, 29, 12. Dirnagl. U .; Iadecola, C .; Moskowitz, MA TINS 1999, 22, 391.].

Therefore, the present inventors synthesized a novel phenylamidine compound having better antioxidant activity than the conventional antioxidant, and the phenylamidine compound of the present invention is more excellent in antioxidant activity than the conventional antioxidant compound, is easy to manufacture, and is toxic. The present invention was completed by knowing the small amount.

An object of the present invention is a phenylamidine derivative and a pharmaceutically acceptable salt having an antioxidant activity, and for the treatment of anti-aging, cancer, diabetes, and epilepsy using the derivative as an active ingredient, and for the treatment of neurodegenerative diseases It is to provide a composition.

In order to achieve the above object,

The present invention provides phenylamidine derivatives having oxidative activity and pharmaceutically acceptable salts thereof.

The present invention also provides a pharmaceutical composition for the treatment of anti-aging, cancer, diabetes and epilepsy and for the treatment of neurodegenerative diseases using the derivative as an active ingredient.

Hereinafter, the present invention will be described in detail.

The present invention includes phenylamidine derivatives represented by the following formula (1) and pharmaceutically acceptable salts thereof.

(Wherein

X ₁ and X ₂ are each independently hydrogen or a linear or branched alkyl group composed of C _{1 to} C ₉ saturated or unsaturated carbons;

Y is CH ₂ or CH═CH (C═O);

Ar represents a phenyl group, a thienyl group or a pyrrolyl group, wherein Ar has no substituent or includes an alkyl group or an alcohol group or a halogen substituent of C _{1 to} C ₃ .

Preferably,

X ₁ and X ₂ are each independently hydrogen, a branched alkyl group containing C _{1 to} C ₉ saturated or unsaturated carbon;

Y is CH ₂ or CH═CH (C═O);

Ar is a phenyl group, thienyl group or pyrrolyl group.)

If more specifically illustrate the phenylamidine derivative represented by the formula (1) as follows.

(1) N- (4- {4- [3- (3,5-di-t-butyl-4-hydroxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -ti Offen-2-carboxamidine,

(2) N- (4- {4- [3- (4-hydroxy-3,5-dimethoxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -thiophene-2 Carboxamidine,

(3) N- [4- (4- {3- [3,5-bis- (1-ethyl-propoxy) -4-hydroxy-phenyl] -acryloyl} -piperazin-1-yl- Phenyl] -thiophene-2-carboxamidine,

(4) N- (4- {4- [3- (4-hydroxy-3,5-dipropoxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -thiophene- 2-carboxamidine,

(5) N- (4- {4- [3- (3,5-dibutoxy-4-hydroxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -thiophene-2 Carboxamidine,

(6) N- (4- {4- [3- (4-hydroxy-3,5-bispentyloxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -thiophene- 2-carboxamidine,

(7) N- (4- {4- [3- (3,5-bis-hexyloxy-4-hydroxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -ti Offen-2-carboxamidine,

(8) N- (4- {4- [3- (3,5-bis-heptyloxy-4-hydroxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -thiophene -2-carboxamidine,

(9) N- (4- {4- [3- (4-hydroxy-3,5-bis-nonyloxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -thiophene -2-carboxamidine,

(10) N- (4- {4- [3- (3,5-di-t-butyl-4-hydroxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -1H Pyrrole-2-carboxamidine,

(11) N- (4- {4- [3- (3,5-dibutoxy-4-hydroxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -1H-pyrrole- 2-carboxamidine,

(12) N- (4- {4- [3- (4-hydroxy-3,5-bis-pentyloxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -1H- Pyrrole-2-carboxamidine,

(13) N- (4- {4- [3- (3,5-bis-heptyloxy-4-hydroxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -1H- Pyrrole-2-carboxamidine,

(14) N- (4- {4- [3- (4-hydroxy-3,5-bis-nonyloxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -1H- Pyrrole-2-carboxamidine,

(15) N- (4- {4- [3- (3,5-di-t-butyl-4-hydroxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -ti Offen-2-carboxamidine hydrochloride,

(16) N- [4- (4- {3- [3,5-bis- (1-ethyl-propoxy) -4-hydroxy-phenyl] -acryloyl} -piperazin-1-yl- Phenyl] -thiophene-2-carboxamidine hydrochloride,

(17) N- (4- {4- [3- (3,5-dibutoxy-4-hydroxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -thiophene-2 Carboxamidine hydrochloride,

(18) N- (4- {4- [3- (3,5-bis-hexyloxy-4-hydroxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -ti Offen-2-carboxamidine hydrochloride,

(19) N- (4- {4- [3- (4-hydroxy-3,5-bis-nonyloxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -thiophene -2-carboxamidine hydrochloride,

(20) N-{(4- {4- (3,5-di-t-butyl-4-hydroxybenzyl) -piperazin-1-yl] -phenyl} -thiophene-2-carboxamidine ,

(21) N- (4- {4- [3,5-bis- (1-ethyl-propoxy) -4-hydroxy-benzyl] -piperazin-1-yl} -phenyl) -thiophene-2 Carboxamidine,

(22) N- {4- [4- (3,5-dibutoxy-4-hydroxy-benzyl) -piperazin-1-yl] -phenyl} -thiophene-2-carboxamidine,

(23) N- {4- [4- (4-hydroxy-3,5-bis-pentyloxy-benzyl) -piperazin-1-yl] -phenyl} -thiophene-2-carboxamidine,

(24) N- {4- [4- (3,5-di-t-butyl-4-hydroxy-benzyl) -piperazin-1-yl] -phenyl} -1 H-pyrrole-2-carboxami Dean,

(25) N- {4- [4- (3,5-dibutoxy-4-hydroxy-benzyl) -piperazin-1-yl] -phenyl} -1H-pyrrole-2-carboxamidine,

(26) N- {4- [4- (4-hydroxy-3,5-bis-pentyloxy-benzyl) -piperazin-1-yl] -phenyl} -1 H-pyrrole-2-carboxamidine .

The present invention includes not only phenylamidine derivatives represented by the general formula (1) and pharmaceutically acceptable salts thereof, but also all possible solvates and hydrates that can be prepared therefrom.

The compound of formula 1 of the present invention may be used in the form of a pharmaceutically acceptable salt, and as the salt, an acid addition salt formed by a pharmaceutically acceptable free acid is useful. Organic acids and inorganic acids may be used as the free acid, and hydrochloric acid, hydrogen bromide, phosphoric acid, sulfuric acid, sodium hydrogen sulfate, nitric acid, carbonic acid, or tartaric acid may be used as the inorganic acid, and formic acid and gestice may be used as the organic acid. Acids, lactobionic acid, salicylic acid, acetylsalicylic acid, methanesulfonic acid, p-toluenesulfonic acid, acetic acid, trifluoroacetic acid, maleic acid, succinic acid, oxalic acid, benzoic acid, tartaric acid, fumaric acid, manderic acid, propionic acid ), Citric acid, lactic acid, glycolic acid, glycolic acid, gluconic acid, galacturonic acid, glutamic acid, glutaric acid, glucuronic acid, Aspartic acid, ascorbic acid, carbonic acid, vanic acid, or hydroiodic acid can be used.

The acid addition salts according to the invention are dissolved in conventional methods, for example, by dissolving a compound of formula 1 in an excess of aqueous acid solution and using the water miscible organic solvent, such as methanol, ethanol, acetone or acetonitrile. It can be prepared by precipitation.

Equivalent amounts of the compound of formula (1) and acid or alcohol (eg, glycol monomethyl ether) in water can be heated and then the mixture is evaporated to dryness or the precipitated salts can be prepared by suction filtration.

Bases can also be used to make pharmaceutically acceptable metal salts. An alkali metal or alkaline earth metal salt is obtained by, for example, dissolving a compound in an excess of alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the insoluble compound salt, and evaporating and drying the filtrate. At this time, it is pharmaceutically suitable to prepare sodium, potassium or calcium salt as the metal salt. Corresponding silver salts are also obtained by reacting alkali or alkaline earth metal salts with a suitable negative salt (eg, silver nitrate).

In the present invention, the phenylamidine derivative represented by Chemical Formula 1 may be prepared by conventional chemical synthesis or extraction from plants, and the present invention is not limited thereto.

In one embodiment, the phenylamidine derivative of the present invention, as represented by Scheme 1 below,

Reducing the compound of formula 2 in the presence of a catalyst to prepare a compound of formula 3 (step 1); And

The compound of Chemical Formula 3 is reacted with the compound of Chemical Formula 4 to prepare a phenylamidine derivative of Chemical Formula 1 (Step 2).

(Wherein X ₁ , X ₂ , Y and Ar are as defined in Formula 1 above.)

Step 1 is to reduce the compound of formula (2) to obtain a compound of formula (3), it can be used all reduction reactions commonly used in the art. For example, ethanol is used as a solvent, iron and hydrochloric acid are added, followed by stirring at room temperature to perform a reduction reaction.

Step 2 is to react the compound of Formula 3 with the compound of Formula 4, in order to easily carry out the reaction it is preferred to react after protecting the hydroxyl group (-OH) of the compound of Formula 3 using a protecting group. The protecting group may use a protecting group commonly used in the art, and preferably a trimethylsilyl group. The hydroxy group is protected and then reacted with a compound of formula 4 in the presence of a solvent and then reacted with a deprotecting agent to prepare a compound of formula 1. The deprotecting agent may be used conventional, preferably tetrabutylammonium fluoride (tetrabutylammonium fluoride) is used.

In addition, the present invention is an anti-aging, cancer, diabetes and epilepsy treatment, and dementia, Parkinson's disease, stroke, Huntington containing the phenylamidine derivative represented by the formula (1) and a pharmaceutically acceptable salt thereof as an active ingredient It includes a pharmaceutical composition for treating neurodegenerative diseases.

The phenylamidine derivatives of the present invention exhibit antioxidant activity. Specifically, when the phenylamidine derivative of the present invention was administered to the rat brain homogenate to determine the antioxidant activity, compared to the conventional antioxidant activator BHT (2.09 μM), the N- [4- (4- {3) of the present invention. -[3,5-bis- (1-ethyl-propoxy) -4-hydroxy-phenyl] -acryloyl} -piperazin-1-yl-phenyl] -thiophene-2-carboxamidine hydro The IC ₅₀ value of the chloride (Example 16) was 0.25 μM with an antioxidant activity of about 88% increased and N- (4- {4- [3- (3,5-dibutoxy-4-hydroxy-phenyl) -Acryloyl] -piperazin-1-yl} -phenyl) -thiophene-2-carboxamidine (Example 5), N- (4- {4- [3- (4-hydroxy-3 , 5-bispentyloxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -thiophene-2-carboxamidine (Example 6) and N- (4- {4- [ 3- (3,5-Dibutoxy-4-hydroxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -1H-pyrrole-2-carboxamidine (Example 11) Increased antioxidant activity by about 82%, resulting in better antioxidant activity The pharmaceutical composition consisting of these phenylamidine derivatives is excellent in antioxidant activity and treated with excessive amounts of ROS in the body to prevent aging, cancer, diabetes and epilepsy, dementia, Parkinson's disease, stroke and Huntington's disease. It is effective for the prevention and treatment of neurodegenerative diseases.

The compound of Formula 1 may be administered in various oral and parenteral formulations during clinical administration, and when formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, surfactants, etc., which are commonly used, may be used. It is manufactured by. Solid form preparations for oral administration include tablets, patients, powders, granules, capsules, troches, and the like, which form at least one excipient such as starch, calcium carbonate, water It is prepared by mixing cross, lactose or gelatin. In addition to simple excipients, lubricants such as magnesium styrate talc are also used. Liquid preparations for oral administration include suspensions, solutions, emulsions, or syrups, and include various excipients such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. Can be. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As the non-aqueous solvent and the suspension solvent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerol, gelatin and the like can be used.

In addition, the dosage of the compound represented by Chemical Formula 1 to the human body may vary depending on the age, weight, sex, dosage form, health condition and degree of disease of the patient, and when the weight is 70 kg based on an adult patient Generally, it is 0.01-1000 mg / day, Preferably it is 0.1-500 mg / day, It can also divide and administer once a day to several times at regular time intervals according to a decision of a doctor or a pharmacist.

Hereinafter, the present invention will be described in more detail with reference to Examples.

However, the following examples are illustrative of the present invention, and the content of the present invention is not limited by the examples.

Example 1 N- (4- {4- [3- (3,5-di-t-butyl-4-hydroxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) Preparation of -thiophene-2-carboxamidine

(Step 1) Preparation of 3- (3,5-di-t-butyl-4-hydroxy-phenyl) -1- [4- (4-nitro-phenyl) -piperazin-1-yl] -propenone

3- (3,5-di-t-butyl-4-hydroxy-phenyl) -acrylic acid (2.00 g, 7.24 mmol), 1- (4-nitrophenyl) piperazine (1.50) in a 100 mL round flask g, 7.24 mmol), EDCI (1- (3-dimethylamino propyl) -3-ethylcarbodiimide hydrochloride, 6.94 g, 36.2 mmol) and HOBT (1-hydroxybenzotriazole hydrate, 4.89 g, 36.2 mmol) was dissolved in purified CH ₃ CN and stirred at room temperature for 3 hours. After CH ₃ CN was removed, CH ₂ Cl ₂ was added, washed with NaHCO ₃ , and washed again with brine and water. The obtained organic layer solvent was dried with Na ₂ SO ₄ , filtered and concentrated. Purification by column chromatography (hexane: EtOAc = 2: 1) 3- (3,5-di-t-butyl-4-hydroxy-phenyl) -1- [4- (4-nitro-phenyl) -pipe 2.854 g (85%) of razin-1-yl] -propenone was obtained.

¹ H-NMR (200 Hz, CDCl ₃ ) δ 1.47 (s, 18H, 6CH ₃ ), 3.50-3.55 (m, 4H, 2NCH ₂ ), 3.88-3.91 (m, 4H, 2NCH ₂ ), 5.49 (s, 1H , OH), 6.68 (d, J = 15.3 Hz, 1H, COCH = CH), 6.84 (d, J = 9.4 Hz, 2H, 2ArH), 7.37 (s, 2H, 2ArH), 7.70 (d, J = 15.1 Hz, 1H, ArCH = CH), 8.16 (d, J = 9.4 Hz, 2H, 2ArH);

Melting point 105-106 ° C.

(Step 2) Preparation of 1- [4- (4-amino-phenyl) -piperazin-1-yl] -3- (3,5-di-t-butyl-4-hydroxy-phenyl) -propenone

In a 50 ml round flask, 3- (3,5-di-t-butyl-4-hydroxy-phenyl) -1- [4- (4-nitro-phenyl) -piperazin-1-yl] -propenone ( After dissolving 500 mg, 1.07 mmol) and Fe (powder) (480 mg, 8.59 mmol) in purified EtOH, HCl (1.30 mL, 15.0 mmol) was slowly added and stirred at room temperature for 4 hours. CH ₂ Cl ₂ was added, washed with NaHCO ₃ and again with brine and water. The obtained organic layer solvent was dried with Na ₂ SO ₄ , filtered and concentrated. Purification by column chromatography (hexane: EtOAc = 1: 1) yields 1- [4- (4-amino-phenyl) -piperazin-1-yl] -3- (3,5-di-t-butyl-4 243 mg (52%) of hydroxy-phenyl) -propenone were obtained.

¹ H-NMR (200 Hz, CDCl ₃ ) δ 1.47 (s, 18H, 6CH ₃ ), 3.07 (t, J = 5.1 Hz, 4H, 2NCH ₂ ), 3.84 (bs, 4H, 2NCH ₂ ), 6.67 (d, J = 8.5Hz, 2H, 2ArH), 6.73 (d, J = 14.4Hz, 1H, COCH = CH), 6.84 (d, J = 8.5Hz, 2H, 2ArH), 7.37 (s, 2H, 2ArH), 7.67 (d, J = 14.4 Hz, 1H, ArCH = CH);

Melting point 81-82 ° C.

 (Step 3) N- (4- {4- [3- (3,5-Di-t-butyl-4-hydroxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl)- Preparation of Thiophene-2-Carboxamidine

In a 50 ml round flask, 1- [4- (4-amino-phenyl) -piperazin-1-yl] -3- (3,5-di-t-butyl-4-hydroxy-phenyl) -propenone ( 200 mg, 0.46 mmol) was dissolved in pyridine, and then HMDS (1,1,1,3,3,3-hexamethyldisilazane, 2.30 mL, 10.7 mmol) and trimethylchlorosilane (2.00 mL, 16.1 mmol) were added dropwise. After heated for 3 hours. After cooling to room temperature, the reaction was terminated with water. Extracted with CH ₂ Cl ₂ and washed again with brine and water. The organic layer solvent was removed by removing water with Na ₂ SO ₄ and concentrated by filtration. To the concentrated compound (10 mg, 0.02 mmol) was added methylsulfanyl-2-thienyl methanimine HI salt (11 mg, 0.04 mmol) and purified chloroform (0.50 mL) was used. After dissolution, acetic acid (0.01 ml, 0.13 mmol) was added dropwise and stirred at room temperature for 5 hours. Chloroform was removed and then dissolved in EtOAc, washed with NaHCO ₃ and again with brine and water. TBAF (tetrabutylammonium fluoride, 2ml) was added dropwise to the organic layer, and the mixture was stirred at room temperature for 20 minutes. After completion of the reaction, the compound was washed with brine and water, dried with Na ₂ SO ₄ , filtered and concentrated. Purified by column chromatography (hexane: EtOAc = 1: 1) to give N- (4- {4- [3- (3,5-di-t-butyl-4-hydroxy-phenyl) -acryloyl]- 9 mg (87%) of piperazin-1-yl} -phenyl) -thiophene-2-carboxamidine was obtained.

Obtained N- (4- {4- [3- (3,5-di-t-butyl-4-hydroxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -thiophene- The melting point (mp) and ¹ H NMR of 2-carboxamidine are shown in Table 1 below.

Example 2 N- (4- {4- [3- (4-hydroxy-3,5-dimethoxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -thiophene Preparation of 2-carboxamidine

(Step 1) Preparation of 3- (4-hydroxy-3,5-dimethoxy-phenyl) -1- [4- (4-nitro-phenyl) -piperazin-1-yl] -propenone

 The experiment was carried out in the same manner as in Step 1 of Example 1, to obtain 750 mg (41%) of the title compound.

¹ H-NMR (200 Hz, CDCl ₃ ) δ 3.53 (m, 4H, 2NCH ₂ ), 3.94 (m, 4H, 2NCH ₂ ), 3.95 (s, 6H, 2OCH ₃ ), 5.73 (s, 1H, OH), 6.67 (d, J = 14.7 Hz, 1H, COCH = CH), 6.78 (s, 2H, 2ArH), 6.85 (d, J = 9.4 Hz, 2H, 2ArH), 7.66 (d, J = 14.7 Hz, 1H, ArCH = CH), 8.17 (d, J = 9.4 Hz, 2H, 2ArH);

Melting point 172-173 ° C.

(Step 2) Preparation of 1- [4- (4-amino-phenyl) -piperazin-1-yl] -3- (4-hydroxy-3,5-dimethoxy-phenyl) -propenone

The same procedure as in Step 2 of Example 1 was carried out to obtain 360 mg (55%) of the title compound.

¹ H-NMR (200 Hz, CDCl ₃ ) δ 3.06 (t, J = 5.1 Hz, 4H, 2NCH ₂ ), 3.85 (bs, 4H, 2NCH ₂ ), 3.93 (s, 6H, 2OCH ₃ ), 6.66 (d, J = 8.7Hz, 2H, 2ArH), 6.75 (d, J = 15.5Hz, 1H, COCH = CH), 6.77 (s, 2H, 2ArH), 6.83 (d, J = 8.7Hz, 2H, 2ArH), 7.61 (d, J = 15.5 Hz, 1H, ArCH = CH);

Melting point 168-169 ℃.

(Step 3) N- (4- {4- [3- (4-hydroxy-3,5-dimethoxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -thiophene- Preparation of 2-carboxamidine

 Experiment in the same manner as in step 3 of Example 1 to give 230 mg (72%) of the target.

Obtained N- (4- {4- [3- (4-hydroxy-3,5-dimethoxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -thiophene-2-car Melting point (mp) and ¹ H NMR of copyamidine are shown in Table 1 below.

Example 3 N- [4- (4- {3- [3,5-bis- (1-ethyl-propoxy) -4-hydroxy-phenyl] -acryloyl} -piperazine-1- Preparation of Il-phenyl] -thiophene-2-carboxamidine

(Step 1) 3- [3,5-bis- (1-ethyl-propoxy) -4-hydroxy-phenyl] -1- [4- (4-nitro-phenyl) -piperazin-1-yl] Preparation of Propenones

The same procedure as in Step 1 of Example 1 was carried out to obtain 457 mg (73%) of the title compound.

¹ H-NMR (200 Hz, CDCl ₃ ) δ 0.98 (t, J = 7.3 Hz, 12H, 4CH ₃ ), 1.70 (qui, J = 6.9 Hz, 8H, 4CH ₂ ), 3.52 (bs, 4H, 2CH ₂ ) , 3.89 (bs, 4H, 2CH ₂ ), 4.19 (t, J = 5.7 Hz, 2H, 2CH), 5.82 (s, 1H, OH), 6.65 (d, J = 15.5 Hz, 1H, CHCH), 6.74 ( s, 2H, 2ArH), 6.83 (d, J = 9.4 Hz, 2H, 2ArH), 7.62 (d, J = 15.5 Hz, 1H, CHCH), 8.14 (d, J = 9.4 Hz, 2H, 2ArH).

(Step 2) 1- [4- (4-Amino-phenyl) -piperazin-1-yl] -3- [3,5-bis- (1-ethyl-propoxy) -4-hydroxy-phenyl] Preparation of Propenones

 The experiment was carried out in the same manner as in Step 2 of Example 1, to obtain 383 mg (92%) of the title compound.

¹ H-NMR (200 Hz, CDCl ₃ ) δ 0.97 (t, J = 7.3 Hz, 12H, 4CH ₃ ), 1.70 (qui, J = 6.9 Hz, 8H, 4CH ₂ ), 3.05 (t, J = 5.1 Hz, 4H, 2CH ₂ ), 3.82 (bs, 4H, 2CH ₂ ), 4.18 (qui, J = 5.7 Hz, 2H, 2CH), 6.65 (d, J = 8.9 Hz, 2H, 2ArH), 6.68 (d, J = 15.3 Hz, 1H, CHCH), 6.72 (s, 2H, 2ArH), 6.81 (d, J = 8.7 Hz, 2H, 2ArH), 7.57 (d, J = 15.3 Hz, 1H, CHCH).

(Step 3) N- [4- (4- {3- [3,5-bis- (1-ethyl-propoxy) -4-hydroxy-phenyl] -acryloyl} -piperazin-1-yl Preparation of -phenyl] -thiophene-2-carboxamidine

The experiment was carried out in the same manner as in Step 3 of Example 1, to obtain 213 mg (50%) of the title compound.

Obtained N- [4- (4- {3- [3,5-bis- (1-ethyl-propoxy) -4-hydroxy-phenyl] -acryloyl} -piperazin-1-yl-phenyl] The melting point (mp) and ¹ H NMR of -thiophene-2-carboxamidine are shown in Table 1 below.

Example 4 N- (4- {4- [3- (4-hydroxy-3,5-dipropoxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -ti Preparation of Offen-2-carboxamidine

(Step 1) Preparation of 3- (4-hydroxy-3,5dipropoxy-phenyl) -1- [4- (4-nitro-phenyl) -piperazin-1-yl] -propenone

3- (4-hydroxy-3,5-dipropoxy-phenyl) -acrylic acid (330 mg, 1.14 mmol), 1- (4-nitrophenyl) piperazine (237 mg, 1.14 mmol) in a 100 ml round flask, EDCI (438 mg, 2.28 mmol) and HOBT (309 mg, 2.28 mmol) were added and dissolved in purified CH ₃ CN, followed by overnight reaction at room temperature. CH ₃ CN was removed and then CH ₂ Cl ₂ was added, washed with NaHCO ₃ and again with brine and water. The organic layer solvent was removed with Na ₂ SO ₄ and then concentrated by filtration.

(Step 2) Preparation of 1- [4- (4-amino-phenyl) -piperazin-1-yl] -3- (4-hydroxy-3,5-dipropoxy-phenyl) -propenone

The experiment was carried out in the same manner as in Step 2 of Example 1, to obtain 403 mg (80%) of the title compound.

¹ H-NMR (200 Hz, CDCl ₃ ) δ 1.05 (t, J = 7.3 Hz, 6H, 2CH ₃ ), 1.86 (sex, J = 7.1 Hz, 4H, 2CH ₂ ), 3.05 (t, J = 4.9 Hz, 4H, 2NCH ₂ ), 3.48 (bs, 2H, NH 2), 3.83 (bs, 4H, 2NCH ₂ ), 4.04 (t, J = 6.7 Hz, 4H, 2OCH ₂ ), 5.70 (bs, 1H, ArOH), 6.66 (d, J = 8.9Hz, 2H, 2ArH), 6.71 (d, J = 15.1Hz, 1H, COCH = CH), 6.75 (s, 2H, 2ArH), 6.81 (d, J = 8.9Hz, 2H, 2ArH ), 7.58 (d, J = 15.1 Hz, 1H, ArCH = CH);

Melting point 72-73 ° C.

(Step 3) N- (4- {4- [3- (4-hydroxy-3,5-dipropoxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -thiophene Preparation of 2-carboxamidine

The same procedure as in Step 3 of Example 1 was conducted to obtain 120 mg (38%) of the title compound.

Obtained N- (4- {4- [3- (4-hydroxy-3,5-dipropoxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -thiophene-2- The melting point (mp) and ¹ H NMR of carboxamidine are shown in Table 1 below.

Example 5 N- (4- {4- [3- (3,5-Dibutoxy-4-hydroxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -thiophene Preparation of 2-carboxamidine

(Step 1) Preparation of 3- (3,5-dibutoxy-4-hydroxy-phenyl) -1- [4- (4-nitro-phenyl) -piperazin-1-yl] -propenone

Experiment in the same manner as in Step 1 of Example 1 to obtain 194 mg (67%) of the target.

¹ H-NMR (200 Hz, CDCl ₃ ) δ 0.99 (t, J = 7.3 Hz, 6H, 2CH ₃ ), 1.51 (sex, J = 7.4 Hz, 4H, 2CH ₂ ) 1.83 (qui, J = 7.0 Hz, 4H , 2CH ₂ ), 3.52 (t, J = 5.2 Hz 4H, 2NCH ₂ ), 3.88-3.90 (m, 4H, 2NCH ₂ ), 5.71 (bs, 1H, OH), 6.69 (d, J = 15.3 Hz, 1H , COCH = CH), 6.76 (s, 2H, 2ArH), 6.83 (d, J = 9.5 Hz, 2H, 2ArH), 7.63 (d, J = 15.3 Hz, 1H, ArCH = CH), 8.15 (d, J = 9.5 Hz, 2H, 2 ArH);

Melting point 191-193 ° C.

(Step 2) Preparation of 1- [4- (4-amino-phenyl) -piperazin-1-yl] -3- (3,5-dibutoxy-4-hydroxy-phenyl) -propenone

 Experiment in the same manner as in step 2 of Example 1 to obtain 140 mg (86%) of the target.

¹ H-NMR (200 Hz, CDCl ₃ ) δ 0.99 (t, J = 7.4 Hz, 6H, 2CH ₃ ), 1.51 (sex, J = 7.3 Hz, 4H, 2CH ₂ ), 1.83 (qui, J = 7.0 Hz, 4H, 2CH ₂ ), 3.06 (t, J = 5.1 Hz, 4H, 2NCH ₂ ), 3.84 (bs, 4H, 2NCH ₂ ), 6.67 (d, J = 8.9 Hz, 2H, 2ArH), 6.72 (d, J = 15.5 Hz, 1H, COCH = CH), 6.76 (s, 2H, 2ArH), 6.82 (d, J = 8.9 Hz, 2H, 2ArH), 7.59 (d, J = 15.5 Hz, 1H, ArCH = CH);

Melting point 65-68 ° C.

(Step 3) N- (4- {4- [3- (3,5-Dibutoxy-4-hydroxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -thiophene- Preparation of 2-carboxamidine

The experiment was carried out in the same manner as in Step 3 of Example 1, to obtain 118 mg (74%) of the title compound.

Obtained N- (4- {4- [3- (3,5-Dibutoxy-4-hydroxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -thiophene-2-car Melting point (mp) and ¹ H NMR of copyamidine are shown in Table 1 below.

Example 6 N- (4- {4- [3- (4-hydroxy-3,5-bispentyloxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -ti Preparation of Offen-2-carboxamidine

(Step 1) Preparation of 3- (4-hydroxy-3,5-bis-pentyloxy-phenyl) -1- [4- (4-nitro-phenyl) -piperazin-1-yl] -propenone

The experiment was carried out in the same manner as in Step 1 of Example 2, to obtain a target product.

(Step 2) Preparation of 1- [4- (4-amino-phenyl) -piperazin-1-yl] -3- (4-hydroxy-3,5-bis-pentyloxy-phenyl) -propenone

Experiments were carried out in the same manner as in Step 2 of Example 1, to obtain 927 mg (82%) of the title compound.

¹ H-NMR (200 Hz, CDCl ₃ ) δ 0.94 (t, J = 6.7 Hz, 6H, 2CH ₃ ), 1.34-1.49 (m, 8H, 4CH ₂ ), 1.85 (qui, J = 6.8 Hz, 4H, 2CH ₂ ), 3.06 (t, J = 5.0 Hz, 4H, 2NCH ₂ ), 3.84 (bs, 4H, 2NCH ₂ ), 4.07 (t, J = 6.6 Hz, 4H, 2OCH ₂ ), 6.66 (d, J = 8.7 Hz, 2H, 2ArH), 6.72 (d, J = 15.3Hz, 1H, COCH = CH), 6.75 (s, 2H, 2ArH), 6.82 (d, J = 8.7Hz, 2H, 2ArH), 7.59 (d, J = 14.4 Hz, 1H, ArCH = CH);

Melting point 55-56 ° C.

(Step 3) N- (4- {4- [3- (4-hydroxy-3,5-bis-pentyloxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -tee Preparation of Offen-2-carboxamidine

The experiment was carried out in the same manner as in Step 3 of Example 1, to obtain 285 mg (89%) of the title compound.

Obtained N- (4- {4- [3- (4-hydroxy-3,5-bis-pentyloxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -thiophene-2 The melting point (mp) and ¹ H NMR of -carboxamidine are shown in Table 1 below.

Example 7 N- (4- {4- [3- (3,5-bis-hexyloxy-4-hydroxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) Preparation of -thiophene-2-carboxamidine

(Step 1) Preparation of 3- (3,5-bis-nucleosiloxy-4-hydroxy-phenyl) -1- [4- (4-nitro-phenyl) -piperazin-1-yl] -propenone

 The experiment was carried out in the same manner as in Step 1 of Example 1, to obtain 373 mg (82%) of the title compound.

¹ H-NMR (200 Hz, CDCl ₃ ) δ 0.90 (t, J = 6.7 Hz, 6H, 2CH ₃ ), 1.25-1.46 (m, 12H, 6CH ₂ ), 1.84 (qui, J = 6.9 Hz, 4H, 2CH ₂ ), 3.51 (t, J = 5.3 Hz, 4H, 2NCH ₂ ), 3.90 (bs, 4H, 2NCH ₂ ), 4.07 (t, J = 6.7 Hz, 4H, 2OCH ₂ ), 5.74 (s, 1H, OH ), 6.68 (d, J = 15.5 Hz, 1H, COCH = CH), 6.76 (s, 2H, 2ArH), 6.83 (d, J = 9.4 Hz, 2H, 2ArH), 7.62 (d, J = 15.5 Hz, 1H, ArCH = CH), 8.14 (d, J = 9.4 Hz, 2H, 2ArH);

Melting point 65-67 ° C.

(Step 2) Preparation of 1- [4- (4-amino-phenyl) -piperazin-1-yl] -3- (3,5-bis-nucleosiloxy-4-hydroxy-phenyl) -propenone

Experiment in the same manner as in step 2 of Example 1 yielded 310 mg (91%) of the title compound.

¹ H-NMR (200 Hz, CDCl ₃ ) δ 0.91 (t, J = 6.7 Hz, 6H, 2CH ₃ ), 1.32-1.47 (m, 12H, 6CH ₂ ), 1.84 (qui, J = 6.9 Hz, 4H, 2CH ₂ ), 3.06 (t, J = 5.3 Hz, 4H, 2NCH ₂ ), 3.43 (bs, 2H, NH 2), 3.84 (bs, 4H, 2NCH ₂ ), 4.07 (t, J = 6.7 Hz, 4H, 2OCH ₂ ), 5.74 (bs, 1H, OH), 6.66 (d, J = 8.8 Hz, 2H, 2ArH), 6.71 (d, J = 15.5 Hz, 1H, COCH = CH), 6.75 (s, 2H, 2ArH), 6.82 (d, J = 8.8 Hz, 2H, 2ArH), 7.58 (d, J = 15.5 Hz, 1H, ArCH = CH).

(Step 3) N- (4- {4- [3- (3,5-bis-hexyloxy-4-hydroxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl)- Preparation of Thiophene-2-Carboxamidine

Experiment in the same manner as in Step 3 of Example 1 yielded 270 mg (74%) of the title compound.

Obtained N- (4- {4- [3- (3,5-bis-hexyloxy-4-hydroxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -thiophene- The melting point (mp) and ¹ H NMR of 2-carboxamidine are shown in Table 1 below.

Example 8 N- (4- {4- [3- (3,5-bis-haptyloxy-4-hydroxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl)- Preparation of Thiophene-2-Carboxamidine

(Step 1) Preparation of 3- (3,5-bis-heptyloxy-4-hydroxy-phenyl) -1- [4- (4-nitro-phenyl) -piperazin-1-yl] -propenone

The experiment was carried out in the same manner as in Step 1 of Example 2, to obtain a target product.

(Step 2) Preparation of 1- [4- (4-amino-phenyl) -piperazin-1-yl] -3- (3,5-bis-heptyloxy-4-hydroxy-phenyl) -propenone

The experiment was carried out in the same manner as in Step 2 of Example 1, to obtain 1.03 mg (78%) of the title compound.

¹ H-NMR (200 Hz, CDCl ₃ ) δ 0.89 (t, J = 6.6 Hz, 6H, 2CH ₃ ), 1.30-1.45 (m, 16H, 8CH ₂ ), 1.84 (qui, J = 7.3H, 2OCH ₂ ) , 3.06 (t, J = 4.9 Hz, 4H, 2NCH ₂ ), 3.84 (bs, 4H, 2NCH ₂ ), 4.07 (t, J = 6.6 Hz, 4H, 2OCH ₂ ), 6.66 (d, J = 8.5 Hz, 2H, 2ArH), 6.72 (d, J = 15.3Hz, 1H, COCH = CH), 6.75 (s, 2H, 2ArH), 6.83 (d, J = 8.5Hz, 2H, 2ArH), 7.59 (d, J = 14.4 Hz, 1H, ArCH = CH);

Melting point 114-115 ° C.

(Step 3) N- (4- {4- [3- (3,5-Bis-haptyloxy-4-hydroxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -ti Preparation of Offen-2-carboxamidine

The experiment was carried out in the same manner as in Step 3 of Example 1, to obtain 133 mg (63%) of the title compound.

Obtained N- (4- {4- [3- (3,5-bis-haptyloxy-4-hydroxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -thiophene-2 The melting point (mp) and ¹ H NMR of -carboxamidine are shown in Table 1 below.

Example 9 N- (4- {4- [3- (4-hydroxy-3,5-bis-nonyloxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl)- Preparation of Thiophene-2-Carboxamidine

(Step 1) Preparation of 3- (4-hydroxy-3,5-bis-nonyloxy-phenyl) -1- [4- (4-nitro-phenyl) -piperazin-1-yl] -propenone

The experiment was carried out in the same manner as in Step 1 of Example 1, to obtain 525 mg (72%) of the title compound.

¹ H-NMR (200 Hz, CDCl ₃ ) δ 0.88 (t, J = 6.5 Hz, 6H, 2CH ₃ ), 1.27-1.45 (m, 24H, 10CH ₃ ), 1.83 (qui, J = 7.0 Hz, 4H, 2CH ₂ ), 3.51 (t, J = 5.2 Hz, 4H, 2NCH ₂ ), 3.90 (t, J = 5.2 Hz, 4H, 2NCH ₂ ), 4.06 (t, J = 6.6 Hz, 4H, 2OCH ₂ ), 5.75 ( s, 1H, OH), 6.69 (d, J = 15.5 Hz, 1H, COCH = CH), 6.76 (s, 2H, 2ArH), 6.82 (d, J = 9.4 Hz, 2H, 2ArH), 7.62 (d, J = 15.5 Hz, 1H, ArCH = CH), 8.14 (d, J = 9.4 Hz, 2H, 2ArH);

Melting point 103-105 캜.

(Step 2) Preparation of 1- [4- (4-amino-phenyl) -piperazin-1-yl] -3- (4-hydroxy-3,5-bis-nonyloxy-phenyl) -propenone

Experiments were carried out in the same manner as in Step 2 of Example 1, to obtain 390 mg (82%) of the title compound.

¹ H-NMR (200 Hz, CDCl ₃ ) δ 0.88 (t, J = 6.5 Hz, 6H, 2CH ₃ ), 1.27-1.45 (m, 24H, 10CH ₃ ), 1.83 (qui, J = 7.0 Hz, 4H, 2CH ₂ ), 3.05 (t, J = 5.2 Hz, 4H, 2NCH ₂ ), 3.46 (bs, 2H, NH 2), 3.83 (bs, 4H, 2NCH ₂ ), 4.06 (t, J = 6.6 Hz, 4H, 2OCH ₂ ), 5.70 (bs, 1H, OH), 6.64 (d, J = 8.9 Hz, 2H, 2ArH), 6.71 (d, J = 15.1 Hz, 1H, COCH = CH), 6.75 (s, 2H, 2ArH), 6.82 (d, J = 8.9 Hz, 2H, 2ArH), 7.59 (d, J = 15.1 Hz, 1H, ArCH = CH);

Melting point 110-111 ° C.

(Step 3) N- (4- {4- [3- (4-hydroxy-3,5-bis-nonyloxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -ti Preparation of Offen-2-carboxamidine

Experimentation was carried out in the same manner as in Step 3 of Example 1, to obtain 360 mg (79%) of the title compound.

N- (4- {4- [3- (4-hydroxy-3,5-bis-nonyloxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -thiophene-2 obtained The melting point (mp) and ¹ H NMR of -carboxamidine are shown in Table 1 below.

Example 10 N- (4- {4- [3- (3,5-di-t-butyl-4-hydroxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) Preparation of -1H-pyrrole-2-carboxamidine

In a 50 ml round flask, 1- [4- (4-amino-phenyl) -piperazin-1-yl] -3- (3,5-di-t-butyl-4-hydroxy-phenyl) -propenone ( 600 mg, 1.38 mmol) was dissolved in pyridine, and then HMDS (1,1,1,3,3,3-hexamethyldisilazane, 2.90 mL, 13.8 mmol) and trimethylchlorosilane (2.60 mL, 20.7 mmol) were added dropwise. Then heated for 3 hours. After cooling to room temperature, the reaction was terminated with water. Extracted with CH ₂ Cl ₂ and washed again with brine and water. The organic layer solvent was removed by removing water with Na ₂ SO ₄ and concentrated by filtration. To the concentrated compound (699 mg, 1.38 mmol) was added 1H-2-azolyl-methylsulfanyl methanimine HI salt (406 mg, 1.52 mmol) and purified 2-propanol ( 10 ml) and then stirred at room temperature for one day. 2-propanol was removed and then dissolved in EtOAc and washed with NaHCO ₃ and again with brine and water. TBAF (tetrabutylammonium fluoride, 2 ml) was added dropwise to the organic layer, and the mixture was stirred at room temperature for 20 minutes. After completion of the reaction, the compound was washed with brine and water, dried with Na ₂ SO ₄ , filtered and concentrated. Purified by column chromatography (MeOH: EtOAc = 1:10) to give N- (4- {4- [3- (3,5-di-t-butyl-4-hydroxy-phenyl) -acryloyl]- 527 mg (72%) of piperazin-1-yl} -phenyl) -1H-pyrrole-2-carboxamidine was obtained.

N- (4- {4- [3- (3,5-di-t-butyl-4-hydroxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -1H-pyrrole obtained Melting point (mp) and ¹ H NMR of -2-carboxamidine are shown in Table 1 below.

Example 11 N- (4- {4- [3- (3,5-Dibutoxy-4-hydroxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -1H- Preparation of Pyrrole-2-carboxamidine

The experiment was carried out in the same manner as in Step 3 of Example 10, to obtain 102 mg (22%) of the title compound.

N- (4- {4- [3- (3,5-Dibutoxy-4-hydroxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -1H-pyrrole-2- obtained The melting point (mp) and ¹ H NMR of carboxamidine are shown in Table 1 below.

Example 12 N- (4- {4- [3- (4-hydroxy-3,5-bis-pentyloxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl)- Preparation of 1H-pyrrole-2-carboxamidine

In the same manner as in Example 3, Step 3, 492 mg (73%) of the title compound was obtained.

Obtained N- (4- {4- [3- (4-hydroxy-3,5-bis-pentyloxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -1H-pyrrole- The melting point (mp) and ¹ H NMR of 2-carboxamidine are shown in Table 1 below.

Example 13 N- (4- {4- [3- (3,5-bis-heptyloxy-4-hydroxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl)- Preparation of 1H-pyrrole-2-carboxamidine

The experiment was carried out in the same manner as in Step 3 of Example 10, to obtain 504 mg (66%) of the title compound.

Obtained N- (4- {4- [3- (3,5-bis-heptyloxy-4-hydroxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -1H-pyrrole- The melting point (mp) and ¹ H NMR of 2-carboxamidine are shown in Table 1 below.

Example 14 N- (4- {4- [3- (4-hydroxy-3,5-bis-nonyloxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl)- Preparation of 1H-pyrrole-2-carboxamidine

The experiment was carried out in the same manner as in Step 3 of Example 10, to obtain 261 mg (62%) of the title compound.

Obtained N- (4- {4- [3- (4-hydroxy-3,5-bis-nonyloxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -1H-pyrrole- The melting point (mp) and ¹ H NMR of 2-carboxamidine are shown in Table 1 below.

Example 15 N- (4- {4- [3- (3,5-Di-t-butyl-4-hydroxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) Preparation of -thiophene-2-carboxamidine hydrochloride

N- (4- {4- [3- (3,5-di-t-butyl-4-hydroxy-phenyl) -acryloyl] -piperazin-1-yl of Example 1 in a 100 ml round flask. } -Phenyl) -thiophene-2-carboxamidine (200 mg, 1.07 mmol) was dissolved in purified MeOH, and then stirred at room temperature for 1 hour while slowly adding HCl gas. After removing MeOH, it was recrystallized with MeOH and ether to give N- (4- {4- [3- (3,5-di-t-butyl-4-hydroxy-phenyl) -acryloyl] -piperazine-1 173 mg (77%) of -yl} -phenyl) -thiophene-2-carboxamidine hydrochloride were obtained.

Obtained N- (4- {4- [3- (3,5-di-t-butyl-4-hydroxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -thiophene- Melting point (mp) and ¹ H NMR of 2-carboxamidine hydrochloride are shown in Table 1 below.

Example 16 N- [4- (4- {3- [3,5-bis- (1-ethyl-propoxy) -4-hydroxy-phenyl] -acryloyl} -piperazine-1- Preparation of Il-phenyl] -thiophene-2-carboxamidine hydrochloride

112 mg (100%) of the title compound was obtained using the compound of Example 3 in the same manner as in Step 4 of Example 1.

Obtained N- [4- (4- {3- [3,5-bis- (1-ethyl-propoxy) -4-hydroxy-phenyl] -acryloyl} -piperazin-1-yl-phenyl] The melting point (mp) and ¹ H NMR of -thiophene-2-carboxamidine hydrochloride are shown in Table 1 below.

Example 17 N- (4- {4- [3- (3,5-Dibutoxy-4-hydroxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -thiophene Preparation of 2-Carboxamidine Hydrochloride

In the same manner as in Example 4, Step 4, 94 mg (76%) of the title compound was obtained using the compound of Example 5.

Obtained N- (4- {4- [3- (3,5-Dibutoxy-4-hydroxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -thiophene-2-car Melting point (mp) and ¹ H NMR of copyamidine hydrochloride are shown in Table 1 below.

Example 18 N- (4- {4- [3- (3,5-bis-hexyloxy-4-hydroxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) Preparation of -thiophene-2-carboxamidine hydrochloride

130 mg (73%) of the title compound was obtained using the compound of Example 7 in the same manner as in Step 4 of Example 1.

Obtained (N- (4- {4- [3- (3,5-bis-hexyloxy-4-hydroxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -thiophene Melting point (mp) and ¹ H NMR of -2-carboxamidine hydrochloride are shown in Table 1 below.

Example 19 N- (4- {4- [3- (4-hydroxy-3,5-bis-nonyloxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl)- Preparation of Thiophene-2-Carboxamidine Hydrochloride

270 mg (98%) of the title compound was obtained using the compound of Example 9 in the same manner as in Step 4 of Example 1.

N- (4- {4- [3- (4-hydroxy-3,5-bis-nonyloxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -thiophene-2 obtained Melting point (mp) and ¹ H NMR of -carboxamidine hydrochloride are shown in Table 1 below.

Example 20 N-{(4- {4- (3,5-di-t-butyl-4-hydroxybenzyl) -piperazin-1-yl] -phenyl} -thiophene-2-carbox Preparation of Amidine

(Step 1) Preparation of 2,6-di (t-butyl) -4- [4- (4-nitrophenyl) piperazinomethyl] phenol

2,6-di (t-butyl) -4-hydroxymethylphenol (1.00 g, 4.23 mmol) and 1- (4-nitrophenyl) piperazine (1.14 g, 5.50 mmol) were added to a 100 mL round flask. Was dissolved by addition. Triphenylphosphine (1.89g, 7.19mmol) and diethyl azodicarboxylate (1.10ml, 7.19mmol) in THF was dissolved in THF using a needle (double ended needle). Added and stirred for 20 minutes. After removing THF, CH ₂ Cl ₂ was added, washed with NaHCO ₃ , and washed again with brine and water. The organic layer solvent was removed by removing water with Na ₂ SO ₄ and concentrated by filtration. Purification by column chromatography (hexane: EtOAc = 5: 1) yielded 1.217 g (68%) of 2,6-di (t-butyl) -4- [4- (4-nitrophenyl) piperazinomethyl] phenol. Got it.

¹ H-NMR (200 Hz, CDCl ₃ ) δ 1.45 (s, 18H, 6CH ₃ ), 2.58 (t, J = 5.1 Hz, 4H, 2NCH ₂ ), 3.43 (t, J = 5.1 Hz, 4H, 2NCH ₂ ) , 3.48 (s, 2H, ArCH ₂ ), 5.16 (s, 1H, ArOH), 6.81 (d, J = 9.4 Hz, 2H, 2ArH), 7.11 (s, 2H, 2ArH), 8.11 (d, J = 9.4 Hz, 2H, 2ArH);

Melting point 171-172 ° C.

(Step 2) Preparation of 4- [4- (4-aminophenyl) piperazinomethyl] -2,6-di (t-butyl) phenol

The experiment was carried out in the same manner as in Step 2 of Example 1, to obtain 520 mg (56%) of the title compound.

¹ H-NMR (200 Hz, CDCl ₃ ) δ 1.45 (s, 18H, 6CH ₃ ), 2.62 (t, J = 10.0 Hz, 4H, 2NCH ₂ ), 3.05-3.10 (m, 6H, 2NCH ₂ , NH ₂ ) , 3.52 (s, 2H, ArCH ₂ ), 5.28 (bs, 1H, ArOH), 6.65 (d, J = 8.9 Hz, 2H, 2ArH), 6.82 (d, J = 8.9 Hz, 2H, 2ArH), 7.12 ( s, 2H, 2ArH); mp 173-174 ° C.

(Step 3) N-{(4- {4- (3,5-Di-t-butyl-4-hydroxybenzyl) -piperazin-1-yl] -phenyl} -thiophene-2-carboxamimi Dean's Preparation

Experimentation was carried out in the same manner as in Step 3 of Example 1, to obtain 99 mg (39%) of the title compound.

The melting point (mp) and ¹ H NMR of the obtained 2,6-di (t-butyl) -4-4- [4-imino (2-cyenyl) methylaminophenyl] piperazinomethylphenol are shown in the following table. 1 is shown.

Example 21 N- (4- {4- [3,5-bis- (1-ethyl-propoxy) -4-hydroxy-benzyl] -piperazin-1-yl} -phenyl) -thiophene Preparation of 2-carboxamidine

(Step 1) Preparation of 2,6-bis- (1-ethyl-propoxy) -4- [4- (4-nitro-phenyl) -piperazin-1-ylmethyl] -phenol

2,6-bis- (1-ethyl-propoxy) -4-hydroxymethyl-phenol (760 mg, 2.56 mmol) and triphenylphosphine (1.89 g, 7.19 mmol) were added to a 100 mL round flask. THF (20 mL) was added to dissolve. N-bromosuccinimide (N-bromosuccinimide, NBS, 502 mg, 2.82 mmol) was slowly added at -20 ° C. After stirring for 5 minutes, 1- (4-nitrophenyl) piperazine (1- (4-nitrophenyl) piperazine, 1.06g, 5.13mmol) was added thereto, and the temperature was slowly raised to room temperature and stirred again for one day. After completion of the reaction, 20 ml of ether was added, followed by filtration, and ether and THF were removed. CH ₂ Cl ₂ was added thereto, washed with brine and water, and the organic layer solvent was removed with Na ₂ SO ₄ , filtered and concentrated. Purified by column chromatography (hexane: EtOAc = 4: 1) to give 2,6-bis- (1-ethyl-propoxy) -4- [4- (4-nitro-phenyl) -piperazin-1-ylmethyl 813 mg (65%) of] -phenol were obtained.

¹ H-NMR (200 Hz, CDCl ₃ ) δ 0.97 (t, J = 7.5 Hz, 12H, 4CH ₃ ), 1.70 (qui, J = 7.0 Hz, 8H, 4CH ₂ ), 2.57 (t, J = 5.1 Hz, 4H, 2NCH ₂ ), 3.42 (t, J = 5.1 Hz, 4H, 2NCH ₂ ), 3.45 (s, 2H, ArCH ₂ ), 4.16 (qui, J = 5.8 Hz, 2H, 2OCH), 5.53 (bs, 1H , ArOH), 6.54 (s, 2H, 2ArH), 6.81 (d, J = 9.4 Hz, 2H, 2ArH), 8.11 (d, J = 9.4 Hz, 2H, 2ArH);

Melting point 100-101 ° C.

(Step 2) Preparation of 4- [4- (4-amino-phenyl) -piperazin-1-ylmethyl] -2,6-bis- (1-ethyl-propoxy) -phenol

The experiment was carried out in the same manner as in Step 2 of Example 1, to obtain 690 mg (99%) of the title compound.

¹ H-NMR (200 Hz, CDCl ₃ ) δ 0.97 (t, J = 7.4 Hz, 12H, 4CH ₃ ), 1.70 (qui, J = 6.7 Hz, 8H, 4CH ₂ ), 2.59 (t, J = 5.0 Hz, 4H, 2NCH ₂ ), 3.06 (t, J = 5.0 Hz, 4H, 2NCH ₂ ,), 3.45 (s, 2H, ArCH ₂ ), 4.16 (qui J = 5.8 Hz, 2H, 2OCH), 6.55 (s, 2H , 2ArH), 6.65 (d, J = 9.6 Hz, 2H, 2ArH), 6.82 (d, J = 9.6 Hz, 2H, 2ArH).

(Step 3) N- (4- {4- [3,5-Bis- (1-ethyl-propoxy) -4-hydroxy-benzyl] -piperazin-1-yl} -phenyl) -thiophene- Preparation of 2-carboxamidine

Experiment in the same manner as in Step 3 of Example 1 yielded 280 mg (87%) of the title compound.

Obtained N- (4- {4- [3,5-bis- (1-ethyl-propoxy) -4-hydroxy-benzyl] -piperazin-1-yl} -phenyl) -thiophen-2-car Melting point (mp) and ¹ H NMR of copyamidine are shown in Table 1 below.

Example 22 of N- {4- [4- (3,5-dibutoxy-4-hydroxy-benzyl) -piperazin-1-yl] -phenyl} -thiophene-2-carboxamidine Produce

(Step 1) Preparation of 2,6-dibutoxy-4- [4- (4-nitro-phenyl) -piperazin-1-ylmethyll] -phenol

The experiment was carried out in the same manner as in Step 5 of Example 16, to obtain 624 mg (73%) of the title compound.

¹ H-NMR (200 Hz, CDCl ₃ ) δ 0.98 (t, J = 7.3 Hz, 6H, 2CH ₃ ), 1.49 (sex, J = 7.5 Hz, 4H, 2CH ₂ ), 1.81 (qui, J = 6.9 Hz, 4H, 2CH ₂ ), 2.57 (t, J = 5.1 Hz, 4H, 2NCH ₂ ), 3.42 (t, J = 5.1 Hz, 4H, 2NCH ₂ ), 3.45 (s, 2H, ArCH ₂ ), 4.04 (t, J = 6.7 Hz, 4H, 20CH ₂ ), 5.47 (bs, 1H, ArOH), 6.56 (s, 2H, 2ArH), 6.81 (d, J = 9.4 Hz, 2H, 2ArH), 8.11 (d, J = 9.4 Hz, 2H, 2ArH);

Melting point 114-115 ° C.

(Step 2) Preparation of 4- [4- (4-amino-phenyl) -piperazin-1-ylmethyl] -2,6-dibutoxy-phenol

The experiment was carried out in the same manner as in Step 2 of Example 1, to obtain 753 mg (81%) of the title compound.

¹ H-NMR (200 Hz, CDCl ₃ ) δ 0.98 (t, J = 7.4 Hz, 6H, 2CH ₃ ), 1.49 (sex, J = 7.5 Hz, 4H, 2CH ₂ ), 1.81 (qui, J = 6.9 Hz, 4H, 2CH ₂ ), 2.57 (t, J = 5.0 Hz, 4H, 2NCH ₂ ), 3.42 (t, J = 5.0 Hz, 4H, 2NCH ₂ ,), 3.45 (s, 2H, ArCH ₂ ), 4.04 (t , J = 6.7 Hz, 4H, 20CH ₂ ), 5.24 (bs, 1H, ArOH), 6.57 (s, 2H, 2ArH), 6.64 (d, J = 9.0 Hz, 2H, 2ArH), 6.81 (d, J = 9.0 Hz, 2H, 2 ArH);

Melting point mp 100-101 ° C.

(Step 3) Preparation of N- {4- [4- (3,5-dibutoxy-4-hydroxy-benzyl) -piperazin-1-yl] -phenyl} -thiophene-2-carboxamidine

The experiment was carried out in the same manner as in Step 3 of Example 1, to obtain 253 mg (79%) of the title compound.

Melting point of the obtained N- {4- [4- (3,5-dibutoxy-4-hydroxy-benzyl) -piperazin-1-yl] -phenyl} -thiophene-2-carboxamidine (mp ) And ¹ H NMR are shown in Table 1 below.

Example 23 N- {4- [4- (4-hydroxy-3,5-bis-pentyloxy-benzyl) -piperazin-1-yl] -phenyl} -thiophene-2-carboxami Dean's Preparation

(Step 1) Preparation of 4- [4- (4-nitro-phenyl) -piperazin-1-ylmethyl] -2,6-bis-pentyloxy-phenol

The experiment was carried out in the same manner as in Step 5 of Example 16, to obtain 937 mg (60%) of the title compound.

¹ H-NMR (200 Hz, CDCl ₃ ) δ 0.93 (t, J = 6.5 Hz, 6H, 2CH ₃ ), 1.26-1.47 (m, 8H, 4CH ₂ ), 1.82 (qui, J = 7.0Hz, 4H, 2CH ₂ ), 2.57 (t, J = 4.9 Hz, 4H, 2NCH ₂ ), 3.40-3.45 (m, 6H, 2NCH ₂ and ArCH ₂ ), 4.03 (t, J = 6.6 Hz, 4H, 2OCH ₂ ), 5.45 ( bs, 1H, ArOH), 6.56 (s, 2H, 2ArH), 6.80 (d, J = 9.5 Hz, 2H, 2ArH), 8.11 (d, J = 9.5 Hz, 2H, 2ArH);

Melting point 94-95 ° C.

(Step 2) Preparation of 4- [4- (4-amino-phenyl) -piperazin-1-ylmethyl] -2,6-bis-pentyloxy-phenol

The same procedure as in Step 2 of Example 1 was carried out to obtain 766 mg (86%) of the title compound.

¹ H-NMR (200 Hz, CDCl ₃ ) δ 0.93 (t, J = 6.5 Hz, 6H, 2CH ₃ ), 1.26-1.47 (m, 8H, 4CH ₂ ), 1.82 (qui, J = 7.0Hz, 4H, 2CH ₂ ), 2.89 (t, J = 4.8 Hz, 4H, 2NCH ₂ ), 3.06 (t, J = 4.8 Hz, 4H, 2NCH ₂ ), 3.46 (s, 2H, ArCH ₂ ), 4.04 (t, J = 6.5 Hz, 4H, 20CH ₂ ), 5.24 (bs, 1H, ArOH), 6.57 (s, 2H, 2ArH), 6.64 (d, J = 9.0Hz, 2H, 2ArH), 6.81 (d, J = 9.0Hz, 2H , 2ArH);

Melting point 131-132 ° C.

(Step 3) N- {4- [4- (4-hydroxy-3,5-bis-pentyloxy-benzyl) -piperazin-1-yl] -phenyl} -thiophene-2-carboxamidine Manufacture

The experiment was carried out in the same manner as in Step 3 of Example 1, to obtain 273 mg (73%) of the title compound.

Melting point of the obtained N- {4- [4- (4-hydroxy-3,5-bis-pentyloxy-benzyl) -piperazin-1-yl] -phenyl} -thiophene-2-carboxamidine (mp) and ¹ H NMR are shown in Table 1 below.

Example 24 N- {4- [4- (3,5-Di-t-butyl-4-hydroxy-benzyl) -piperazin-1-yl] -phenyl} -1 H-pyrrole-2-car Preparation of Copy Amidine

The experiment was carried out in the same manner as in Step 3 of Example 20, to obtain 88 mg (36%) of the title compound.

Of obtained N- {4- [4- (3,5-di-t-butyl-4-hydroxy-benzyl) -piperazin-1-yl] -phenyl} -1H-pyrrole-2-carboxamidine Melting point (mp) and ¹ H NMR are shown in Table 1 below.

Example 25 N- {4- [4- (3,5-Dibutoxy-4-hydroxy-benzyl) -piperazin-1-yl] -phenyl} -1 H-pyrrole-2-carboxamidine Manufacture

Experiment in the same manner as in Step 3 of Example 20 to obtain 220 mg (53%) of the target substance.

Melting point of N- {4- [4- (3,5-dibutoxy-4-hydroxy-benzyl) -piperazin-1-yl] -phenyl} -1H-pyrrole-2-carboxamidine obtained ( mp) and ¹ H NMR are shown in Table 1 below.

Example 26 N- {4- [4- (4-hydroxy-3,5-bis-pentyloxy-benzyl) -piperazin-1-yl] -phenyl} -1 H-pyrrole-2-carbox Preparation of Amidine

Experiment in the same manner as in Step 3 of Example 20 to obtain 263 mg (55%) of the title compound.

Melting of the obtained N- {4- [4- (4-hydroxy-3,5-bis-pentyloxy-benzyl) -piperazin-1-yl] -phenyl} -1H-pyrrole-2-carboxamidine The points (mp) and ¹ H NMR are shown in Table 1 below.

The melting point and ¹ H NMR of the phenylamidine compound represented by Chemical Formula 1 obtained according to the preparation method of the embodiment of the present invention are shown in Table 1 below.

Table 1 (continued)

Table 1 (continued)

Table 1 (continued)

Table 1 (continued)

Table 1 (continued)

On the other hand, the phenylamidine compound represented by Formula 1 according to the present invention can be formulated in various forms according to the purpose. The following are some examples of formulation methods containing the compound represented by Formula 1 according to the present invention as an active ingredient, but the present invention is not limited thereto.

Preparation Example 1 Tablet (Direct Pressurization)

After sifting 5.0 mg of the active ingredient, 14.1 mg of lactose, 0.8 mg of crospovidone USNF, and 0.1 mg of magnesium stearate were mixed and pressed to prepare a tablet.

Preparation Example 2 Tablet (Wet Assembly)

After sifting 5.0 mg of the active ingredient, 16.0 mg of lactose and 4.0 mg of starch were mixed. 0.3 mg of polysorbate 80 was dissolved in pure water and then an appropriate amount of this solution was added and then atomized. After drying, the fine particles were sieved and mixed with 2.7 mg of colloidal silicon dioxide and 2.0 mg of magnesium stearate. The granules were pressed to make tablets.

Preparation Example 3 Powder and Capsule

After sifting 5.0 mg of the active ingredient, it was mixed with 14.8 mg of lactose, 10.0 mg of polyvinyl pyrrolidone, and 0.2 mg of magnesium stearate. The mixture was prepared using a suitable apparatus. Filled in 5 gelatin capsules.

<Example 4> Injection

Injectables were prepared by containing 100 mg as the active ingredient, followed by the addition of 180 mg of mannitol, 26 mg of Na ₂ HPO ₄ .12H ₂ O and 2974 mg of distilled water.

Experimental Example 1 Antioxidant Activity Test

1. Preparation of brain homogenate

SD rats (males, 10-12 weeks old) were single headed and brains were removed quickly and homogenized after addition of 10 mM Tris-HCl buffer (pH 7.4) containing 150 mM KCl (pH 7.4). The homogenized brain mixture was centrifuged at 2,200 rpm and 4 ° C for 10 minutes, and only the supernatant was taken to determine the exact amount of protein through protein assay and then stored at -20 ° C.

2. Lipid Peroxidation Assay

Dispense 250 μl of brain homogenate (5 mg protein / ml), 10 μl of test substance, and 20 μl of buffer solution in 96-well microplates, and incubate at 37 ° C. for 20 min in 20 μM FeCl ₂ and 250 μM. 10 μl each of ascorbic acid was added thereto and then incubated at 37 ° C. for 30 minutes. 50 μl of 35% HClO ₄ was added to stop the reaction. The microplates were centrifuged at 2000 rpm and 4 ° C. for 10 min to transfer 240 μl of the supernatant to 96-well microplates, followed by TBA (thiobarbituric acid; 5 mg). / Ml in 50% acetic acid) was added in 120 mu l. The microplates were reacted at 80 ° C. for 1 hour and then cooled at room temperature, and then absorbance was measured at 520 nm of TBARS (thiobarbituric acid reactive substances (MDA)) produced by the reaction.

Quantitative reaction curves of TBARS generated using tetraethoxypropane, a reactant of TBA, were used to calculate the amount of reaction product MDA of the test substance, and the inhibitory effect of the test drug was calculated by Equation 1 below. In addition, 50% inhibition concentration (IC ₅₀ ) was calculated by calculating the dose response curve.

(Wherein

A represents the concentration (nmol) of the control protein (MDA / mg),

B represents the concentration (nmol) of test protein (MDA / mg).)

Therefore, a smaller value of IC ₅₀ indicates a stronger antioxidant activity. The experimental results are shown in Table 2 below.

Experimental compound Lipid Peroxidation Assay (IC ₅₀ ) N- (4- {4- [3- (3,5-Di-t-butyl-4-hydroxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -thiophene-2 Carboxamidine (Example 1) 0.51 μM N- [4- (4- {3- [3,5-bis- (1-ethyl-propoxy) -4-hydroxy-phenyl] -acryloyl} -piperazin-1-yl-phenyl]- Thiophene-2-Carboxamidine (Example 3) 0.47 μM N- (4- {4- [3- (4-hydroxy-3,5-dipropoxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -thiophene-2-car Copyamidine (Example 4) 0.49 μM N- (4- {4- [3- (3,5-Dibutoxy-4-hydroxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -thiophene-2-carbox Amidine (Example 5) 0.38 μM N- (4- {4- [3- (4-hydroxy-3,5-bispentyloxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -thiophene-2-car Copyamidine (Example 6) 0.37 μM N- (4- {4- [3- (3,5-Bis-hexyloxy-4-hydroxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -thiophene-2 Carboxamidine (Example 7) 0.40 μM N- (4- {4- [3- (3,5-Dibutoxy-4-hydroxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -1H-pyrrole-2-car Copyamidine (Example 11) 0.37 μM N- (4- {4- [3- (4-hydroxy-3,5-bis-pentyloxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -1H-pyrrole-2 Carboxamidine (Example 12) 0.45 μM N- [4- (4- {3- [3,5-bis- (1-ethyl-propoxy) -4-hydroxy-phenyl] -acryloyl} -piperazin-1-yl-phenyl]- Thiophene-2-Carboxamidine Hydrochloride (Example 16) 0.25 μM N- (4- {4- [3- (3,5-Bis-hexyloxy-4-hydroxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -thiophene-2 Carboxamidine hydrochloride (Example 18) 0.40 μM BHT 2.09 μM

As shown in Table 2 , the smaller the IC ₅₀ value, the stronger the antioxidant activity, compared to the conventional antioxidant activator BHT (2.09 μM) N- [4- (4- {3- [3, 5-bis- (1-ethyl-propoxy) -4-hydroxy-phenyl] -acryloyl} -piperazin-1-yl-phenyl] -thiophene-2-carboxamidine hydrochloride (Example The IC ₅₀ value of 16) was 0.25 μM, increasing the antioxidant activity by about 88%, and N- (4- {4- [3- (3,5-dibutoxy-4-hydroxy-phenyl) -acryloyl ] -Piperazin-1-yl} -phenyl) -thiophene-2-carboxamidine (Example 5), N- (4- {4- [3- (4-hydroxy-3,5-bis Pentyloxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -thiophene-2-carboxamidine (Example 6) and N- (4- {4- [3- (3 , 5-dibutoxy-4-hydroxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -1H-pyrrole-2-carboxamidine (Example 11) has a weak antioxidant activity. 82% increase in antioxidant activity Because of this, it can be useful for anti-aging and neurodegenerative diseases such as cancer, diabetes, epilepsy, stroke, Parkin's disease, dementia, Huntington's disease.

Experimental Example 2 Oral Acute Toxicity in Rats

On the other hand, in order to determine the acute toxicity of the compound of Formula 1 was carried out the following experiment.

Acute toxicity test was performed using 6-week-old SPF SD rats. Two animals per group were suspended orally administered at a dose of 10 mg / kg / 15 ml, each of the compounds obtained in Examples 1-26, suspended in 0.5% methylcellulose solution.

After administration of the test substance, mortality, clinical symptoms, and changes in body weight were examined. Hematological and hematological examinations were performed. Necropsy was performed to visually observe abdominal and thoracic organ abnormalities.

As a result, all animals treated with test substance showed no clinical symptoms and no dead animals, and no toxic changes were observed in weight change, blood test, blood biochemical test, autopsy findings. As a result, all of the tested compounds did not show toxic changes up to 10 mg / kg in rats, and the minimum oral dose (LD ₅₀ ) was determined to be a safe substance of 100 mg / kg or more.

As described above, since the phenylamidine derivative represented by Formula 1 according to the present invention has excellent antioxidant activity, anti-aging, anti-cancer, diabetes, epilepsy treatment or prevention of neurodegenerative diseases such as stroke, Parkin's disease, dementia, etc. And for treatment.

Claims (5)

A phenylamidine derivative represented by the following formula (1) and a pharmaceutically acceptable salt thereof. Formula 1 (Wherein X ₁ and X ₂ are each independently a linear or branched alkoxy group containing a saturated or unsaturated carbon of C ₃ to C ₉ ; Y is CH ₂ or CH═CH (C═O); Ar represents a phenyl group, thienyl group or pyrrolyl group, wherein Ar does not include a substituent or includes an alkyl or alkoxy group having _{1 to 3} carbon atoms or a halogen substituent.) 2. A compound according to claim 1, wherein X ₁ and X ₂ are each independently a branched alkyl group containing a saturated or unsaturated carbon of C ₃ to C ₇ , Y is CH ₂ or CH═CH (C═O), and Ar is a thienyl group. Or a phenylamidine derivative characterized in that it is a pyrrolyl group and a pharmaceutically acceptable salt thereof. According to claim 1, wherein the phenylamidine derivative represented by the formula (1) (3) N- [4- (4- {3- [3,5-bis- (1-ethyl-propoxy) -4-hydroxy-phenyl] -acryloyl} -piperazin-1-yl- Phenyl] -thiophene-2-carboxamidine, (4) N- (4- {4- [3- (4-hydroxy-3,5-dipropoxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -thiophene- 2-carboxamidine, (5) N- (4- {4- [3- (3,5-dibutoxy-4-hydroxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -thiophene-2 Carboxamidine, (6) N- (4- {4- [3- (4-hydroxy-3,5-bispentyloxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -thiophene- 2-carboxamidine, (7) N- (4- {4- [3- (3,5-bis-hexyloxy-4-hydroxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -ti Offen-2-carboxamidine, (8) N- (4- {4- [3- (3,5-bis-heptyloxy-4-hydroxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -thiophene -2-carboxamidine, (9) N- (4- {4- [3- (4-hydroxy-3,5-bis-nonyloxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -thiophene -2-carboxamidine, (11) N- (4- {4- [3- (3,5-dibutoxy-4-hydroxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -1H-pyrrole- 2-carboxamidine, (12) N- (4- {4- [3- (4-hydroxy-3,5-bis-pentyloxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -1H- Pyrrole-2-carboxamidine, (13) N- (4- {4- [3- (3,5-bis-heptyloxy-4-hydroxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -1H- Pyrrole-2-carboxamidine, (14) N- (4- {4- [3- (4-hydroxy-3,5-bis-nonyloxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -1H- Pyrrole-2-carboxamidine, (16) N- [4- (4- {3- [3,5-bis- (1-ethyl-propoxy) -4-hydroxy-phenyl] -acryloyl} -piperazin-1-yl- Phenyl] -thiophene-2-carboxamidine hydrochloride, (17) N- (4- {4- [3- (3,5-dibutoxy-4-hydroxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -thiophene-2 Carboxamidine hydrochloride, (18) N- (4- {4- [3- (3,5-bis-hexyloxy-4-hydroxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -ti Offen-2-carboxamidine hydrochloride, (19) N- (4- {4- [3- (4-hydroxy-3,5-bis-nonyloxy-phenyl) -acryloyl] -piperazin-1-yl} -phenyl) -thiophene -2-carboxamidine hydrochloride, (21) N- (4- {4- [3,5-bis- (1-ethyl-propoxy) -4-hydroxy-benzyl] -piperazin-1-yl} -phenyl) -thiophene-2 Carboxamidine, (22) N- {4- [4- (3,5-dibutoxy-4-hydroxy-benzyl) -piperazin-1-yl] -phenyl} -thiophene-2-carboxamidine, (23) N- {4- [4- (4-hydroxy-3,5-bis-pentyloxy-benzyl) -piperazin-1-yl] -phenyl} -thiophene-2-carboxamidine, (25) N- {4- [4- (3,5-dibutoxy-4-hydroxy-benzyl) -piperazin-1-yl] -phenyl} -1H-pyrrole-2-carboxamidine, and (26) N- {4- [4- (4-hydroxy-3,5-bis-pentyloxy-benzyl) -piperazin-1-yl] -phenyl} -1 H-pyrrole-2-carboxamidine Phenylamidine derivatives and pharmaceutically acceptable salts thereof. An antioxidant comprising the phenylamidine derivative of claim 1 and a pharmaceutically acceptable salt thereof as an active ingredient. Anti-aging, anti-cancer, diabetes, epilepsy, or stroke, Parkinson's disease, Huntington's disease and dementia with the phenylamidine derivative of claim 1 as an active ingredient thereof. Pharmaceutical composition for the treatment of degenerative diseases.