KR100542324B1 - 2,2-Dimethyl-3-hydroxy-4-alkoxy-6-alkyl amino benzopyran libraries by parallel synthesis on the solid-phase - Google Patents

Abstract

The present invention uses a novel 2,2-dimethyl-3-hydroxy-4-alkoxy-6-alkyl amino benzopyran derivative and a solid phase equilibrium synthesis method, one of the combinatorial chemical synthesis techniques, to synthesize the above novel compounds with high efficiency. The present invention relates to a method for use as an agent for the prevention and treatment of diseases caused by the promotion of lipid peroxidation or the accumulation of oxides because the novel compounds exhibit excellent lipid peroxidation inhibitory effects.

Combination Chemical Synthesis, Solid Equilibrium Synthesis, Antioxidants, Inhibition of Lipid Peroxidation, Cerebral Nerve Disease, Hypertension, Diabetes, Pharmacological Effects, Benzopyran, Library

Description

2,2-dimethyl-3-hydroxy-4-alkoxy-6-alkyl amino benzopyran libraries by parallel using solid phase equilibrium synthesis synthesis on the solid-phase}

The present invention uses a novel 2,2-dimethyl-3-hydroxy-4-alkoxy-6-alkyl amino benzopyran derivative and a solid phase equilibrium synthesis method, one of the combinatorial chemical synthesis techniques, to synthesize the above novel compounds with high efficiency. The present invention relates to a method for use as an agent for the prevention and treatment of diseases caused by the promotion of lipid peroxidation or the accumulation of oxides because the novel compounds exhibit excellent lipid peroxidation inhibitory effects.

Injury or death of neurons is known to be a major cause of various neurological diseases, including stroke, brain trauma, Alzheimer's disease and Parkinson's disease [GJ Zoppo et al. , Drugs 54 , 9 ( 1997 ); I. Sziraki et al. , Neurosci. 85 , 1101 ( 1998 )]. Many factors are known to damage neurons, and the representative ones include increasing iron concentrations, increasing free radicals, and increasing oxides in neurons [MP Mattson et al. , Methods Cell Biol. 46 , 187 ( 1995 ); Y. Goodman et al. , Brain Res. 706 , 328 ( 1996 ).

When iron concentration increases in nerve cells, oxygen free radicals such as NO are formed. When excessively large amounts of oxygen free radicals are generated, lipid peroxidation is promoted and oxidizing substances are increased and accumulated in cells. Oxidized substances accumulated in the cells not only cause inflammatory diseases such as arthritis, myocardial infarction, and dementia in addition to the neurological diseases described above, but also damage organs by endotoxins due to tissue damage or bacterial infection during reperfusion in ischemic diseases. It is known to cause damage to various acute tissues or organs.

Therefore, by developing a substance that prevents damage to nerve cells by increasing iron concentration in the nerve cells, prevents lipid peroxidation, and inhibits NO production by endotoxins, various diseases caused by nerve cell damage or death are developed. Can be prevented or treated. Currently, antioxidants have been reported to mitigate neuronal damage and death by iron in neurons, and efforts are underway to develop drugs to prevent neuronal damage caused by oxidative stress [Y. Zhang et al. , J. Cereb. Blood Flow Metab. 13 , 378 ( 1993 ).

Benzopyran skeleton and natural products and synthetic compounds have antioxidant activity and are widely known and widely used as a basic skeletal structure in the development of compounds possessing pharmacological effects such as neurological diseases, hypertensions and diabetes treatments.

On the other hand, building a benzopyran library with various substituents using combinatorial chemical synthesis can be useful in the search for bio-hit compounds and lead compounds in the early stages of drug development. have.

In particular, if various substituents can be introduced into the molecule, and if there is hydroxy at adjacent carbons, the efficient construction of a large-scale and concentrated library by combinatorial chemical synthesis technology by introducing various functional groups such as esters and ethers can be useful for the search for leading substances. This is important in terms of strategies for securing diversity.

Combinatorial chemical synthesis is a new synthesis of new material development, whereas conventional organic synthesis synthesizes a single compound in a single reaction, whereas combinatorial chemical synthesis involves a greater variety of compounds. It is a high-efficiency chemical synthesis method that can synthesize at the same time or automate the multi-step synthesis process. The introduction of combinatorial chemical synthesis facilitates the search for new structures of biological hit compounds and lead compounds, as well as optimization of their structure and activity.

In addition, in the combination chemical synthesis technology, most of the reaction process is performed on a solid support, it is possible to automate continuous multistage reactions and reaction processes, and the separation and purification process of the product is very simple, so that high throughput screening (HTS) is achieved. The advantage is that it is possible.

Although combinatorial chemical synthesis is a new synthetic technology that overcomes the inefficiency and inefficiency of the existing synthetic technology, there are several reasons why it could not be easily applied to the field of organic synthesis. One of the typical causes is that most of the chemical reactions on the solid support use excessive amounts of reaction reagents, which in some cases cause unwanted side reactions and may be used depending on the physical properties of the solid support used. Due to the limitation of the solvent, the choice of reaction conditions is extremely narrow. In the field of solid-phase combinatorial chemical synthesis, Merrifield resin and Wang resin are widely used as solid supports, but these supports have very swelling effects in polar solvents such as alcohol and water. It is low and thus places a lot of restrictions on the choice of solvents required for the reaction. Therefore, in order to synthesize various derivatives using solid phase chemical reaction, selection of solid support and reaction reagents and search of reaction conditions are important factors.

The inventors have found that benzopyran derivatives are useful for synthesizing compounds possessing pharmacological effects such as antioxidants, neurological diseases, hypertension and diabetes treatment, and efficiently construct a benzopyran library using combinatorial chemical synthesis on solid phase. Research efforts have been made to develop ways to do this. As a result, in general, in the classical chemical reaction of the solution phase, the target compound is synthesized by the reaction, the post-reaction treatment purification, and the structure confirmation operation at every stage, and the present inventors use a combination chemical synthesis technique using a solid phase equilibrium synthesis method. After carrying out the reaction in three stages in succession, and then through easy reaction treatment process, a large amount of target compound 2,2-dimethyl-3-hydroxy-4-alkoxy-6-alkyl amino benzo in a short time and efficiently The pyran library was obtained in high yield to complete the present invention.

It is therefore an object of the present invention to provide novel 2,2-dimethyl-3-hydroxy-4-alkoxy-6-alkyl amino benzopyran derivatives that are effective for drug synthesis.

In addition, the present invention is easy to automate the continuous multi-step reaction process and chemical structural analysis of the final compound by using the solid phase equilibrium synthesis method, 2,2-dimethyl-3 by solid phase equilibrium synthesis method having the advantages of excellent production yield Another object is to provide a method for synthesizing a -hydroxy-4-alkoxy-6-alkylamino benzopyran derivative.

In addition, since the present invention has excellent lipid peroxidation inhibitory activity, 2,2-dimethyl-3-hydroxy-4-alkoxy-6-alkyl amino benzopyran as an agent for the prevention and treatment of diseases caused by the promotion of lipid peroxidation or the accumulation of oxides It is another object to provide the use of derivatives.

The present invention is characterized by the 2,2-dimethyl-3-hydroxy-4-alkoxy-6-alkyl amino benzopyran derivative represented by the following general formula (4a).

In the formula (4a), R ^1a represents an allyl group, benzyl group or substituted benzyl group, naphthylmethyl group, or phenethyl group; R ² represents a C ₁ to C ₁₀ alkyl group, benzyl group or substituted benzyl group, or — (CH ₂ ) _n -phenyl group, where n is an integer of 2 or 3; In this case, the substituted benzyl group represents a benzyl group in which 1 to 4 substituents selected from a halogen atom, a nitro group, a C _{1 to} C ₁₀ alkyl group, a C _{1 to} C ₁₀ alkoxy group, and a C _{1 to} C ₁₀ haloalkyl group are substituted.

On the other hand, 2,2-dimethyl-3-hydroxy-4-alkoxy-6-alkyl amino benzopyran derivatives represented by the following general formula (4), which is the object of the present invention, are optical isomer compounds, respectively, by conventional separation methods. It can also be separated by isomer compounds.

Referring to the present invention in more detail as follows.

The present invention is a solid phase equilibrium of a 2,2-dimethyl-3-hydroxy-4-alkoxy-6-alkyl amino benzopyran derivative represented by the following formula (4) with the novel benzopyran derivative described above rather than a solution phase chemical reaction. It is characterized by the manufacturing method by the combination chemical synthesis technique which can be efficiently constructed using the synthesis method, and the use of the new compound as a preventive and therapeutic agent for diseases caused by the promotion of lipid peroxidation or the accumulation of oxides. do.

A reaction for preparing a 2,2-dimethyl-3-hydroxy-4-alkoxy-6-alkyl amino benzopyran derivative represented by the following general formula (4) according to the present invention by a combination chemical synthesis technique is shown briefly. Same as 1.

In the above scheme 1, R ¹ is C ₁ ~C ₁₀ alkyl group, an allyl group, a benzyl group or substituted benzyl group, a naphthylmethyl group, or the pen represents an ethyl group; R ² represents a C ₁ to C ₁₀ alkyl group, benzyl group or substituted benzyl group, or — (CH ₂ ) _n -phenyl group, where n is an integer of 2 or 3; Wherein the substituted benzyl group represents a benzyl group in which 1 to 4 substituents are selected from a halogen atom, a nitro group, a C _{1 to} C ₁₀ alkyl group, a C _{1 to} C ₁₀ alkoxy group and a C _{1 to} C ₁₀ haloalkyl group; X represents a halogen atom; Ⓟ represents a solid support in the form of a high molecular polymer selected from polystyrene-divinylbenzene, methacrylic acid-dimethylacrylamide and hydroxy methacrylic acid.

As the reaction intermediate synthesized in the preparation method according to the present invention, 2,2-dimethyl-3-hydroxy-4-alkoxy-6-alkyl amino carbamate resin represented by the formula (3) is also an optical isomer. It can also be separated into individual isomeric compounds.

The preparation method of the present invention according to Scheme 1 includes the following processes: by selectively introducing an alkyl substituent to a nitrogen atom of benzopyran linked by a carbamate linker represented by Formula (1), A first step of synthesizing the N-alkyl substituted carbamate resin represented by (2); Meta-chloroperbenzoic acid ( m- CPBA) and an alcohol are added to the compound represented by the above formula (2) to simultaneously perform the epoxidation reaction and the alkoxy addition reaction to the 2,2-dimethyl-3- represented by the above formula (3). A second step reaction of synthesizing resin in the form of hydroxy-6-alkyl amino benzopyran; And the compound represented by the formula (3) is desorbed using a dichloromethane solution containing trifluoroacetic acid (TFA) or an organic solvent containing an organic acid and is represented by the formula (4). Third step reaction to synthesize 3-hydroxy-4-alkoxy-6-alkyl amino benzopyran derivatives.

According to the experimental results of the present inventors, when the benzopyran resin of the carbamate form on the solid support represented by the formula (1) is continuously subjected to the N-alkylation reaction and the hydroxy alkoxylation reaction, the classical solution phase reaction By greatly reducing the purification process as compared to the above it was possible to synthesize a variety of 2,2-dimethyl-3-hydroxy-4-alkoxy-6-alkyl ami benzopyran derivatives in a short time.

Referring to the reaction process according to the invention, the composition of the solvent system and the selection range of the reaction conditions in detail.

In the present invention, an organic solvent having excellent swelling effect of Wang resin or Merrifield resin is used as the solvent. In the first stage reaction, dimethyl sulfoxide (DMSO) or tetrahydrofuran (THF) is used as a solvent, wherein about 3 equivalents of lithium t -butoxide (LiO t Bu) is used as the base and alkyl bromide is used as the electrophile. It is good to use, Preferably it is excellent in economy to use in about 2 equivalents.

The second step in the reaction of meta-chloroperbenzoic acid (m -CPBA) In the m -CPBA in order to suppress the addition reaction of 3-chlorobenzoic acid by-product produced during the oxidation reaction within 3 equivalents and, in particular before its use the m -CPBA It is preferable to add an oxidizing agent after the excess of alcohols which are alkoxy group precursors are mixed in advance for 15 minutes or more, and it mixes well enough. This is because the alkoxy group is intended to suppress the formation of by-products by allowing the alkoxy group to be absolutely superior to the amount of 3-chlorobenzoic acid produced after the oxidation reaction. Alcohols as alkoxy precursors refer to aliphatic alcohols, for example, methanol, ethanol, iso-propyl alcohol, benzyl alcohol, substituted benzyl alcohol, or aliphatic alcohol in the side chain and the amine group and carboxylic acid moiety as a protecting group. Refers to protected amino acids.

In the third step reaction, a dechlorination reaction using a dichloromethane solution containing trifluoroacetic acid (TFA) or an organic solvent containing an organic acid is carried out so that the present invention is represented by the general formula (4). Obtain a dimethyl-3-hydroxy-4-alkoxy-6-alkyl amino benzopyran library.

In addition, to confirm the generation of the 2,2-dimethyl-3-hydroxy-4-alkoxy-6-alkyl amino benzopyran library represented by the formula (4) according to the present invention, in the final step after the reaction The target compound desorbed from 2,2-dimethyl-3-hydroxy-4-alkoxy-6-alkyl amino benzopyran resin represented by the formula (3) is purified and separated by flash column chromatography using a multi-column apparatus. Well, the structure was analyzed and confirmed by NMR and Mass spectra. The resins represented by the formulas (1), (2) and (3), which are reaction intermediates, confirmed the progress of the reaction by measuring ATR-FTIR.

On the other hand, the compounds of the present invention can be used as a lipid peroxidation inhibitor due to the excellent activity of inhibiting lipid peroxidation by iron, it can be usefully used as a preventive and therapeutic agent for various diseases caused by lipid peroxidation and the accumulation of oxides have. That is, the compounds of the present invention attack lipids, which are the major components of cell membranes, to produce cytotoxic lipid peroxide or destroy cell membranes, such as cancer, arteriosclerosis, diabetes, stroke, dementia, Parkinson's disease, and aging. It can be used as a drug for preventing or treating a disease.

Accordingly, the present invention contains a 2,2-dimethyl-3-alkylether-4-alkoxy-6-alkylamino benzopyran derivative or a pharmaceutically acceptable salt thereof as an active ingredient, thereby promoting lipid peroxidation or oxidizing It includes a pharmaceutical composition effective for the prevention and treatment of various diseases caused by accumulation.

Pharmaceutically acceptable salts in the present invention can be prepared by conventional methods in the art, for example, salts with inorganic acids such as hydrochloric acid, hydrogen bromide, sulfuric acid, sodium hydrogen sulfate, phosphoric acid, carbonic acid, and the like. Or pharmaceutically acceptable organic compounds, such as formic acid, acetic acid, oxalic acid, benzoic acid, citric acid, tartaric acid, gluconic acid, gestyic acid, fumaric acid, lactobionic acid, salicylic acid, or acetylsalicylic acid (aspirin) It is also possible to form salts of acids, or to react with alkali metal ions such as sodium, potassium, to form their metal salts, or to react with ammonium ions to form another form of a pharmaceutically acceptable salt.

In addition, the pharmaceutical compositions of the present invention are conventional non-toxic pharmaceutically acceptable carriers in 2,2-dimethyl-3-alkylether-4-alkoxy-6-alkylamino benzopyran derivatives or pharmaceutically acceptable salts thereof, Adjuvants, excipients and the like can be added to formulate common oral or parenteral formulations such as tablets, capsules, troches, solutions, suspensions, and the like in the pharmaceutical art. In addition, the dosage of the compound according to the present invention to the human body may vary depending on the age, weight, sex, dosage form, health condition and degree of disease of the patient, and generally based on an adult patient having a weight of 70 kg. It is 0.01-1000 mg / day, and can be dividedly administered once a day to several times at fixed time intervals according to the judgment of a doctor or a pharmacist.

The present invention as described above will be described in more detail based on the following examples, but the present invention is not limited thereto.

Example I: N-alkyl Substitution and Desorption of Olefin Resin 1

I-1) N-methyl Substitution and Desorption Reaction of Olefin Resin 1

Carbamate resin (200.00 mg, 0.11 mmol) in the form of benzopyran represented by the formula (1) was added to dimethylsulfoxide (DMSO) (3 ml), shaken at room temperature for 10 minutes, and dissolved in tetrahydrofuran (THF). Lithium t -butoxide (LiO t Bu) (0.33 mL, 0.33 mmol) was added at a molar concentration, followed by mixing for 20 minutes at the same temperature, followed by addition of iodine methane (MeI) (0.020 mL, 0.33 mmol), followed by 35 Shake and mix for 15 h. After completion of the reaction, the reaction mixture was filtered and washed repeatedly with DMF, MC, MC / MeOH, MeOH to obtain N-methyl carbamate resin (205.7 mg) as a pale brown solid represented by the formula (2a). (ATR-FTIR Analysis; N-methylation carbamate: 1700 cm ^-1 )

The obtained N-methyl carbamate resin (2a) was added to a dichloromethane (5 mL) solution, shaken and trifluoroacetic acid (TFA) (1 mL) was added, followed by shaking at room temperature for 4 hours. After the reaction was completed, the reaction mixture was filtered, the filtrate was washed repeatedly with CH ₂ Cl ₂ and MeOH, and the filtrates were combined and concentrated. Ethyl acetate (3 ml) was added to the concentrated mixture, which was then filtered through SAX resin (strong anion exchange resin) and washed repeatedly with ethyl acetate to remove residual trifluoroacetic acid. The filtrate was concentrated under reduced pressure, and then purified by column chromatography on silica gel under a mixed solvent of hexane / ethyl acetate (4/1, v / v ) to give a pale yellow oil (17.7 mg, resin 1) represented by the formula (5a); yield = 81%) from a loading capacity of 0.55 mmol / g.

^{1 H NMR (300 MHz, CDCl} 3) δ (PPM) 6.67 (d, 1H, J = 8.5Hz), 6.45 (dd, 1H, J = 8.5 Hz, J = 2.7), 6.33 (d, 1H, J = 2.7 Hz), 6.27 (d, 1H, J = 9.7 Hz), 5.62 (d, 1H, J = 9.7 Hz), 2.80 (s, 3H), 1.40 (s, 6H); HRMS 189.1157

I-2) N-ethyl Substitution and Desorption of Olefin Resin 1

Carbamate resin (200.00 mg, 0.11 mmol) in the form of benzopyran represented by the formula (1) was added to dimethylsulfoxide (DMSO) (3 ml), shaken at room temperature for 10 minutes, and dissolved in tetrahydrofuran (THF). Lithium t -butoxide (LiO t Bu) (0.33 mL, 0.33 mmol) was added at a molar concentration, followed by mixing for 20 minutes at the same temperature, followed by addition of iodine ethane (EtI) (0.026 mL, 0.33 mmol), followed by 35 Shake and mix for 15 h. After completion of the reaction, the reaction mixture was filtered and washed repeatedly with DMF, MC, MC / MeOH, MeOH to obtain N-methyl carbamate resin (205.3 mg) as a pale brown solid represented by the formula (2b). (ATR-FTIR Analysis; N-methylation carbamate: 1700 cm ^-1 )

The obtained N-methyl carbamate resin (2b) was added to a dichloromethane (5 mL) solution, shaken, trifluoroacetic acid (TFA) (1 mL) was added, and then shaken at room temperature for 4 hours. After the reaction was completed, the reaction mixture was filtered, the filtrate was washed repeatedly with CH ₂ Cl ₂ and MeOH, and the filtrates were combined and concentrated. Ethyl acetate (3 ml) was added to the concentrated mixture, which was then filtered through SAX resin (strong anion exchange resin) and washed repeatedly with ethyl acetate to remove residual trifluoroacetic acid. The filtrate was concentrated under reduced pressure, and then purified by column chromatography on silica gel under a mixed solvent of hexane / ethyl acetate (4/1, v / v ) to give a pale yellow oil (19.5 mg, Resin 1; yield = 84%) from loading capacity of 0.55 mmol / g.

HRMS 203.2867

I-3) N-2-propyl Substitution and Desorption of Olefin Resin 1

Carbamate resin (200.00 mg, 0.11 mmol) in the form of benzopyran represented by the formula (1) was added to dimethylsulfoxide (DMSO) (3 ml), shaken at room temperature for 10 minutes, and dissolved in tetrahydrofuran (THF). Molar lithium t -butoxide (LiO t Bu) (0.33 mL, 0.33 mmol) was added, followed by mixing for 20 minutes at the same temperature. 2-Iopropane (2-ProI) (0.033 mL, 0.33 mmol) was added and stirred at 35 ° C. for 15 hours to mix. After completion of the reaction, the reaction mixture was filtered and washed repeatedly with DMF, MC, MC / MeOH, MeOH to obtain N-isopropyl carbamate resin (204.5 mg) as a pale brown solid represented by the formula (2c). (ATR-FTIR Analysis; N-methylation carbamate: 1700 cm ^-1 )

The obtained N-isopropyl carbamate resin (2c) was added to a dichloromethane (5 mL) solution, shaken and trifluoroacetic acid (TFA) (1 mL) was added, followed by shaking at room temperature for 4 hours. After the reaction was completed, the reaction mixture was filtered, the filtrate was washed repeatedly with CH ₂ Cl ₂ and MeOH, and the filtrates were combined and concentrated. Ethyl acetate (3 ml) was added to the concentrated mixture, which was then filtered through SAX resin (strong anion exchange resin) and washed repeatedly with ethyl acetate to remove residual trifluoroacetic acid. The filtrate was concentrated under reduced pressure, and then purified by column chromatography on silica gel under a mixed solvent of hexane / ethyl acetate (4/1, v / v ) to give a pale yellow oil (19.4 mg, Resin 1; yield = 79%) from a loading capacity of 0.55 mmol / g.

HRMS 217.3134

I-4) N-butyl Substitution and Desorption of Benzopyran Resin 1

Carbamate resin (200.00 mg, 0.11 mmol) in the form of benzopyran represented by the formula (1) was added to dimethylsulfoxide (DMSO) (3 ml), shaken at room temperature for 10 minutes, and dissolved in tetrahydrofuran (THF). Lithium t -butoxide (LiO t Bu) (0.33 mL, 0.33 mmol) was added at a molar concentration, followed by mixing for 20 minutes at the same temperature, followed by addition of iobutane (BuI) (0.038 mL, 0.33 mmol), followed by 35 Shake and mix for 15 h. After completion of the reaction, the reaction mixture was filtered and washed repeatedly with DMF, MC, MC / MeOH, MeOH to obtain N-butyl carbamate resin (203.9 mg) as a pale brown solid represented by the formula (2d). (ATR-FTIR Analysis; N-methylation carbamate: 1700 cm ^-1 )

The obtained N-butyl carbamate resin (2d) was added to a dichloromethane (5 mL) solution, shaken and trifluoroacetic acid (TFA) (1 mL) was added, followed by shaking at room temperature for 4 hours. After the reaction was completed, the reaction mixture was filtered, the filtrate was washed repeatedly with CH ₂ Cl ₂ and MeOH, and the filtrates were combined and concentrated. Ethyl acetate (3 ml) was added to the concentrated mixture, which was then filtered through SAX resin (strong anion exchange resin) and washed repeatedly with ethyl acetate to remove residual trifluoroacetic acid. The filtrate was concentrated under reduced pressure, and then purified by column chromatography on silica gel under a mixed solvent of hexane / ethyl acetate (4/1, v / v ) to give a pale yellow oil (20.7 mg, resin 1; yield = 80%) from loading capacity of 0.55 mmol / g).

HRMS 231.3409

I-5) N-pentyl Substitution and Desorption of Benzopyran Resin 1

Carbamate resin (200.00 mg, 0.11 mmol) in the form of benzopyran represented by the formula (1) was added to dimethylsulfoxide (DMSO) (3 ml), shaken at room temperature for 10 minutes, and dissolved in tetrahydrofuran (THF). Lithium t -butoxide (LiO t Bu) (0.33 mL, 0.33 mmol) was added at a molar concentration, followed by mixing for 20 minutes at the same temperature, followed by addition of pentane pentane (PenI) (0.043 mL, 0.33 mmol), followed by 35 Shake and mix for 15 h. After completion of the reaction, the reaction mixture was filtered and washed repeatedly with DMF, MC, MC / MeOH, MeOH to obtain N-pentyl carbamate resin (203.9 mg) as a pale brown solid represented by the formula (2e). (ATR-FTIR Analysis; N-methylation carbamate: 1700 cm ^-1 )

The obtained N-pentyl carbamate resin (2e) was added to a dichloromethane (5 mL) solution, shaken, trifluoroacetic acid (TFA) (1 mL) was added, and then shaken at room temperature for 4 hours. After the reaction was completed, the reaction mixture was filtered, the filtrate was washed repeatedly with CH ₂ Cl ₂ and MeOH, and the filtrates were combined and concentrated. Ethyl acetate (3 ml) was added to the concentrated mixture, which was then filtered through SAX resin (strong anion exchange resin) and washed repeatedly with ethyl acetate to remove residual trifluoroacetic acid. The filtrate was concentrated under reduced pressure, and then purified by column chromatography on silica gel under a mixed solvent of hexane / ethyl acetate (4/1, v / v ) to give a pale yellow oil (22.1 mg, resin 1; yield = 80%) from loading capacity of 0.55 mmol / g).

¹ H NMR (300 MHz, CDCl ₃ ) δ (PPM) 6.64 (d, 1H, J = 8.5 Hz), 6.43 (dd, 1H, J = 8.5 Hz, J = 2.8 Hz), 6.31 (d, 1H, J = 2.8 Hz), 6.26 (d, 1H, J = 9.7 Hz), 5.61 (d, 1H, J = 9.7 Hz), 3.04 (t, 2H, 7.1 Hz), 1.60 (m, 2H), 1.39-1.34 ( m, 10H), 0.91 (t, 3H, J = 6.7 Hz); ¹³ C (75 MHz) δ 145.23, 142.08, 131.56, 122.61, 121.91, 116.80, 114.23, 111.29, 75.47, 45.40, 29.32, 29.17, 27.55, 22.49, 14.02; HRMS 245.1780

I-6) N-benzyl Substitution and Desorption of Benzopyran Resin 1

Carbamate resin (200.00 mg, 0.11 mmol) in the form of benzopyran represented by the formula (1) was added to dimethylsulfoxide (DMSO) (3 ml), shaken at room temperature for 10 minutes, and dissolved in tetrahydrofuran (THF). Molar lithium t -butoxide (LiO t Bu) (0.33 mL, 0.33 mmol) was added, followed by mixing for 20 minutes at the same temperature, followed by addition of benzyl bromide (BnBr) (0.039 mL, 0.33 mmol), followed by 35 Shake and mix for 15 h. After completion of the reaction, the reaction mixture was filtered and washed repeatedly with DMF, MC, MC / MeOH, MeOH to obtain N-benzyl carbamate resin (203.9 mg) as a pale brown solid represented by the formula (2f). (ATR-FTIR Analysis; N-methylation carbamate: 1700 cm ^-1 )

The obtained N-benzyl carbamate resin (2f) was added to a dichloromethane (5 mL) solution, shaken, trifluoroacetic acid (TFA) (1 mL) was added, and then shaken at room temperature for 4 hours. After the reaction was completed, the reaction mixture was filtered, the filtrate was washed repeatedly with CH ₂ Cl ₂ and MeOH, and the filtrates were combined and concentrated. Ethyl acetate (3 ml) was added to the concentrated mixture, which was then filtered through SAX resin (strong anion exchange resin) and washed repeatedly with ethyl acetate to remove residual trifluoroacetic acid. The filtrate was concentrated under reduced pressure, and then purified by column chromatography on silica gel under a mixed solvent of hexane / ethyl acetate (4/1, v / v ) to give a pale yellow oil (25.0 mg, resin 1; yield = 85% from a loading capacity of 0.55 mmol / g).

¹ H NMR (300 MHz, CDCl ₃ ) δ (PPM) 7.39-7.26 (m, 5H), 6.64 (d, 1H, J = 8.5 Hz), 6.45 (dd, 1H, J = 8.5 Hz, J = 2.8 Hz ), 6.33 (d, 1H, J = 2.8 Hz), 6.24 (d, 1H, J = 9.7 Hz), 5.60 (d, 1H, J = 9.7 Hz), 4.27 (s, 2H), 1.39 (s, 6H) ); ¹³ C (75 MHz) δ 145.17, 142.19, 139.52, 131.59, 128.55, 127.62, 127.17, 122.58, 121.90, 116.80, 113.88, 111.00, 75.46, 49.34, 27.55; HRMS 265.1467

I-7) N- (2-methyl) benzyl Substitution Reaction (5g) and Desorption of Benzopyran Resin 1

Under the same conditions, a pale yellow oil (24.2 mg, resin 1; yield = 78% from loading capacity of 0.55 mmol / g) represented by the formula (5 g) was obtained.

¹ H NMR (300 MHz, CDCl ₃ ) δ (PPM) 7.33 (s, 1H), 7.21 to 7.17 (m, 3H), 6.66 (d, 1H, J = 8.5 Hz), 6.46 (dd, 1H, J = 8.5 Hz, J = 2.8 Hz, 6.34 (d, 1H, J = 2.8 Hz), 6.25 (d, 1H, J = 9.7 Hz), 5.61 (d, 1H, J = 9.7 Hz), 4.21 (s, 2H ), 2.37 (s, 3 H), 1.40 (s, 6 H); HRMS 279.1623

I-8) N- (4-methyl) benzyl substitution reaction of benzopyran resin 1 (5h) and desorption reaction

Under the same conditions, a pale yellow oil (27.3 mg, resin 1; yield = 87% from loading capacity of 0.55 mmol / g) represented by the formula (5h) was obtained.

¹ H NMR (300 MHz, CDCl ₃ ) δ (PPM) 7.25 (d _AB , 2H, J = 8.0 Hz), 7.14 (d _AB , 1H, J = 8.0 Hz), 6.64 (d, 1H, J = 8.4 Hz ), 6.47 (dd, 1H, J = 8.6 Hz, J = 2.7 Hz), 6.36 (d, 1H, J = 2.7 Hz), 6.24 (d, 1H, J = 9.5 Hz), 5.60 (d, 1H, J = 9.7 Hz), 4.21 (s, 2H), 2.34 (s, 3H), 1.40 (s, 6H); ¹³ C (75 MHz) δ 145.69, 141.30, 136.99, 135.79, 131.61, 129.24, 127.83, 122.52, 121.92, 116.83, 114.57, 111.69; HRMS 279.1623

I-9) N- (4- of Benzopyran Resin 1 t -Butyl) benzyl substitution reaction (5i) and desorption reaction

Under the same conditions, a pale yellow oil (28.7 mg, resin 1; yield = 80% from loading capacity of 0.55 mmol / g) represented by Chemical Formula (5i) was obtained.

¹ H NMR (300 MHz, CDCl ₃ ) δ (PPM) 7.38-7.29 (m, 4H), 6.65 (d, 1H, J = 8.5 Hz), 6.47 (dd, 1H, 8.5 Hz, J = 2.8 Hz), 6.47 (dd, 1H, J = 8.5 Hz, J = 2.8 Hz), 6.36 (d, 1H, J = 2.8 Hz), 6.25 (d, 1H, 9.7 Hz), 5.61 (d, 1H, J = 9.7 Hz) 4.22 (s, 2H), 1.40 (s, 6H), 1.32 (s, 9H); ¹³ C (75 MHz) δ 150.39, 145.84, 141.12, 135.63, 131.64, 127.75, 125.50, 122.51, 121.94, 116.85, 114.74, 111.84, 75.59, 49.66, 34.49, 31.33, 27.59; HRMS 321.2093

I-10) N- (4-fluoro) benzyl substitution of benzopyran resin 1 (5 J ) And desorption reaction

Under the same conditions, a pale yellow oil (26.4 mg, resin 1; yield = 83% from loading capacity of 0.55 mmol / g) represented by the formula (5j) was obtained.

¹ H NMR (300 MHz, CDCl ₃ ) δ (PPM) 7.33 to 7.30 (m, 2H), 7.02 to 6.99 (m, 2H), 6.64 (d, 1H, J = 5.1 Hz), 6.45 (dd, 1H, J = 5.1 Hz, J = 1.7 Hz), 6.34 (d, 1H, J = 1.7 Hz), 6.23 (d, 1H, J = 5.8Hz), 5.61 (d, 1H, J = 5.8 Hz), 4.22 (s , 2H), 1.39 (s, 6H); ¹³ C (75 MHz) δ 163.03, 161.08, 145.61, 141.43, 134.87, 131.70, 129.22, 122.46, 121.95, 116.85, 115.45, 115.28, 114.29, 111.44, 75.56, 48.92, 27.57; HRMS 283.1375

I-11) N- (3-fluoro) benzyl substitution reaction (5k) and desorption reaction of benzopyran resin 1

Under the same conditions, a pale yellow oil (24.8 mg, resin 1; yield = 78% from loading capacity of 0.55 mmol / g) represented by the formula (5k) was obtained.

^{1 H NMR (300 MHz, CDCl} 3) δ (PPM) 6.64 (d, 1H, J = 8.5Hz), 6.43 (dd, 1H, J = 8.5 Hz, J = 2.8), 6.31 (d, 1H, J = 2.8 Hz), 6.26 (d, 1H, J = 9.7 Hz), 5.61 (d, 1H, J = 9.7 Hz), 3.04 (t, 2H, 7.1 Hz), 1.60 (m, 2H), 1.39-1.34 (m , 10H), 0.91 (t, 3H, J = 6.7 Hz); HRMS 283.1372

I-12) N- (2,5-dimethyl) benzyl substitution reaction (5l) and desorption reaction of benzopyran resin 1

Under the same conditions, a pale yellow oil (25.1 mg, resin 1; yield = 77% from loading capacity of 0.55 mmol / g) represented by Chemical Formula (5l) was obtained.

¹ H NMR (300 MHz, CDCl ₃ ) δ (PPM) 7.10 (s, 1H), 6.97 (s, 2H), 6.86 (dd, 1H, J = 5.1 Hz, J = 1.5 Hz), 6.79 (d, 1H , J = 1.5 Hz), 6.67 (d, 1H, J = 5.1 Hz), 6.22 (d, 1H, J = 5.9 Hz), 5.64 (d, 1H, J = 5.9 Hz), 4.21 (s, 2H), 2.22 (s, 3H), 2.11 (s, 3H), 1.40 (s, 6H); HRMS 293.1789

I-13) N- (2-fluoro-4-bromo) benzyl substitution reaction (5m) and desorption reaction of benzopyran resin 1

Under the same conditions, a pale yellow oil (31.4 mg, resin 1; yield = 79% from loading capacity of 0.55 mmol / g) represented by the formula (5m) was obtained.

¹ H NMR (300 MHz, CDCl ₃ ) δ (PPM) 7.28 to 7.21 (m, 3H), 6.63 (d, 1H, J = 5.4 Hz), 6.45 (dd, 1H, J = 5.4 Hz, J = 1.7 Hz ), 6.34 (d, 1H, J = 1.7 Hz), 6.22 (d, 1H, J = 5.8 Hz), 5.61 (d, 1H, J = 5.8 Hz), 4.29 (s, 2H), 1.39 (s, 6H) ); HRMS 361.0478

I-14) N-alkyl Substitution Reaction (5n) and Desorption Reaction of Benzopyran Resin 1

Under the same conditions, a pale yellow oil (28.2 mg, resin 1; yield = 82% from loading capacity of 0.55 mmol / g) represented by the formula (5n) was obtained.

¹ H NMR (300 MHz, CDCl ₃ ) δ (PPM) 7.82-7.79 (m, 4H), 7.48-7.45 (m, 3H), 6.66 (d, 1H, J = 5.1 Hz), 6.54 (dd, 1H, J = 5.1 Hz, J = 1.7 Hz), 6.46 (d, 1H, J = 1.7 Hz), 6.23 (d, 1H, J = 5.9 Hz), 5.61 (d, 1H, J = 5.9 Hz), 4.41 (s , 2H), 1.41 (s, 6H); HRMS 315.1619

I-15) N-Phenyl Substitution Reaction (5o) and Desorption Reaction of Benzopyran Resin 1

Under the same conditions, a pale yellow oil (26.2 mg, resin 1; yield = 85% from loading capacity of 0.55 mmol / g) represented by the formula (5o) was obtained.

¹ H NMR (300 MHz, CDCl ₃ ) δ (PPM) 7.37 to 7.22 (m, 5H), 6.66 (d, 1H, J = 5.1 Hz), 6.61 (d, 1H, J = 9.5 Hz), 6.50 (dd , 1H, J = 5.1 Hz, J = 1.7 Hz, 6.38 (d, 1H, J = 1.7 Hz), 6.33 (dd, 1H, J = 9.5 Hz, J = 3.5 Hz), 6.26 (d, 1H, J = 5.9 Hz), 5.61 (d, 1H, J = 5.9 Hz), 3.87 (dd, 1H, J = 3.5 Hz, J = 0.9 Hz), 1.40 (s, 6H); ¹³ C (75 MHz) δ 145.40, 141.83, 136.86, 131.60, 131.57, 128.53, 127.48, 127.13, 126.30, 122.58, 121.93, 116.82, 114.36, 111.46, 75.51, 47.41, 27.57; HRMS 279.1623

I-16) N- (4-methoxy) benzyl substitution reaction of benzopyran resin 1 (5p) and desorption reaction

Under the same conditions, a pale yellow oil (25.0 mg, resin 1; yield = 77% from loading capacity of 0.55 mmol / g) represented by the formula (5p) was obtained.

HRMS 295.3845

I-17) N- (4-trifluoromethyl) benzyl substitution reaction (5q) and desorption reaction of benzopyran resin 1

Under the same conditions, a pale yellow oil (30.5 mg, resin 1; yield = 81% from loading capacity of 0.55 mmol / g) represented by the formula (5q) was obtained.

HRMS 333.3568

I-18) N- (2,4,6-trimethyl) benzyl substitution (5r) and desorption reaction of benzopyran resin 1

Under the same conditions, a pale yellow oil (26.3 mg, resin 1; yield = 78% from loading capacity of 0.55 mmol / g) represented by the formula (5r) was obtained.

HRMS 307.4397

I-19) N-allyl Substitution Reaction (5s) and Desorption Reaction of Benzopyran Resin 1

Under the same conditions, a pale yellow oil (18.8 mg, resin 1; yield = 77% from loading capacity of 0.55 mmol / g) represented by the formula (5s) was obtained.

¹ H NMR (300 MHz, CDCl ₃ ) δ (PPM) 6.65 (d, 1H, J = 5.1 Hz), 6.51 (dd, 1H, J = 5.1 Hz, J = 1.7 Hz), 6.40 (d, 1H, J = 1.7 Hz), 6.25 (d, 1H, J = 5.9 Hz), 5.95 (m, 1H), 5.61 (d, 1H, J = 5.9 Hz), 5.28 (dd, 1H, J = 10.2 Hz, J = 0.9 Hz), 5.16 (dd, 1H, J = 6.0 Hz, J = 0.9 Hz), 3.72 (d, 2H, J = 2.4 Hz), 1.40 (s, 6H); ¹³ C (75 MHz) δ 145.96, 140.75, 134.91, 131.64, 122.47, 121.92, 116.96, 116.83, 115.00, 112.13, 75.61, 48.36, 27.61; HRMS 215.1319

I-20) N-propargyl Substitution Reaction (5t) and Desorption Reaction of Benzopyran Resin 1

Under the same conditions, a pale yellow oil (17.3 mg, resin 1; yield = 71% from loading capacity of 0.55 mmol / g) represented by the formula (5t) was obtained.

¹ H NMR (300 MHz, CDCl ₃ ) δ (PPM) 6.68 (d, 1H, J = 5.1 Hz), 6.50 (dd, 1H, J = 5.1 Hz, J = 1.7 Hz), 6.38 (d, 1H, J = 1.7 Hz), 6.27 (d, 1H, J = 5.9 Hz), 5.62 (d, 1H, J = 5.9 Hz), 3.88 (d, 2H, 1.5 Hz), 2.21 (t, 1H, 1.5), 1.40 ( s, 6H); HRMS 213.2815

I-21) N-cinnamil substitution reaction (5u) and desorption reaction of benzopyran resin 1

Under the same conditions, a pale yellow oil (23.6 mg, resin 1; yield = 73% from loading capacity of 0.55 mmol / g) represented by the formula (5u) was obtained.

¹ H NMR (300 MHz, CDCl ₃ ) δ (PPM) 7.37 to 7.22 (m, 5H), 6.66 (d, 1H, J = 5.1 Hz), 6.61 (d, 1H, J = 9.5 Hz), 6.50 (dd , 1H, J = 5.1 Hz, J = 1.7 Hz, 6.38 (d, 1H, J = 1.7 Hz), 6.33 (dd, 1H, J = 9.5 Hz, J = 3.5 Hz), 6.26 (d, 1H, J = 5.9 Hz), 5.61 (d, 1H, J = 5.9 Hz), 3.87 (dd, 1H, J = 3.5 Hz, J = 0.9 Hz), 1.40 (s, 6H); ¹³ C (75 MHz) δ 145.40, 141.83, 136.86, 131.60, 131.57, 128.53, 127.48, 127.13, 126.30, 122.58, 121.93, 116.82, 114.36, 111.46, 75.51, 47.41, 27.57; HRMS 291.3965

Example II Hydroxy Alkoxy Addition and Desorption of N-Methyl-Substituted Olefin Resins (2a)

II-1) Hydroxymethoxy Addition and Desorption Reaction of N-Methyl Substituted Olefin Resin (2a)

N-methyl-substituted olefin resin (200.00 mg, 0.11 mmol) represented by the formula (2a) was added to a mixed solution of dichloromethane (3 ml) and methanol (3 ml) and shaken for 30 minutes, followed by meta-chloroperbenzoic acid ( m- CPBA). ) (81 mg, 0.55 mmol) was added at room temperature, followed by reaction at 35 ° C. for 15 hours. After completion of the reaction, the reaction mixture was filtered and washed repeatedly with DMF, MC, MC / MeOH, MeOH to give a resin (208.6 mg) as a pale yellow solid represented by the formula (3-1). The obtained resin (3-1) was added to a dichloromethane (5 mL) solution, shaken, trifluoroacetic acid (TFA) (1 mL) was added, and then shaken at room temperature for 4 hours. After the reaction was completed, the reaction mixture was filtered, the filtrate was washed repeatedly with CH ₂ Cl ₂ and MeOH, and the filtrates were combined and concentrated. Ethyl acetate (3 ml) was added to the concentrated mixture, which was then filtered through SAX resin (strong anion exchange resin) and washed repeatedly with ethyl acetate to remove residual trifluoroacetic acid. The filtrate was concentrated under reduced pressure, and then purified by column chromatography on silica gel under a mixed solvent of hexane / ethyl acetate (4/1, v / v ) to obtain a pale yellow oil (16.3 mg, Resin 1; yield = 63% from loading capacity 0.55 mmol / g) was obtained.

¹ H NMR (300 MHz, CDCl ₃ ) δ (PPM) 6.69 (d, 1H, J = 5.2 Hz), 6.65 (d, 1H, J = 1.7 Hz), 6.58 (dd, 1H, J = 5.2 Hz, J = 1.7 Hz), 4.34 (d, 1H, J = 4.4 Hz), 3.84 (d, 1H, J = 4.4 Hz), 3.49 (s, 3H), 2.82 (s, 3H), 1.43 (s, 3H), 1.26 (s, 3 H); ¹³ C (75 MHz) δ 145.49, 143.00, 121.33, 117.84, 115.50, 111.70, 78.16, 71.99, 55.70, 31.90, 25.68, 20.04; M / S 237.30

II-2) Hydroxyethoxy Addition and Desorption of N-Methyl Substituted Olefin Resin (2a)

The formula (2a) N- methyl substituted olefin resin (200.00 ㎎, 0.11 mmol) represented by dichloromethane (3 ㎖) and ethanol (3 ㎖) into a mixed solution stirs for 30 minutes, meta-chloroperbenzoic acid (m -CPBA ) (81 mg, 0.55 mmol) was added at room temperature, followed by reaction at 35 ° C. for 15 hours. After completion of the reaction, the reaction mixture was filtered and washed repeatedly with DMF, MC, MC / MeOH, MeOH to obtain a resin (207.4 mg) as a pale yellow solid represented by the formula (3-2). The obtained resin (3-2) was added to a dichloromethane (5 mL) solution, shaken and trifluoroacetic acid (TFA) (1 mL) was added, followed by shaking at room temperature for 4 hours. After the reaction was completed, the reaction mixture was filtered, the filtrate was washed repeatedly with CH ₂ Cl ₂ and MeOH, and the filtrates were combined and concentrated. Ethyl acetate (3 ml) was added to the concentrated mixture, which was then filtered through SAX resin (strong anion exchange resin) and washed repeatedly with ethyl acetate to remove residual trifluoroacetic acid. The filtrate was concentrated under reduced pressure, and then purified by column chromatography on silica gel under a mixed solvent of hexane / ethyl acetate (4/1, v / v ) to give a pale yellow oil (16.7 mg, Resin 1; yield = 60% from loading capacity 0.55 mmol / g) was obtained.

¹ H NMR (200 MHz, CDCl ₃ ) δ (PPM) 6.70 to 6.50 (m, 3H), 4.36 (d, 1H, J = 7.4 Hz), 3.82 to 3.67 (m, 3H), 2.81 (s, 3H) , 1.43 (s, 3H), 1.32-1.25 (m, 6H); M / S 251.33

II-3) Hydroxy 2-propoxy addition and desorption reaction of N-methyl substituted olefin resin (2a)

N-methyl-substituted olefin resin (200.00 mg, 0.11 mmol) represented by the formula (2a) was added to a mixed solution of dichloromethane (3 mL) and 2-propanol (3 mL), and shaken for 30 minutes, followed by meta-chloroperbenzoic acid ( m -CPBA) (81 mg, 0.55 mmol) was added at room temperature and then reacted by shaking at 35 to 15 hours. After completion of the reaction, the reaction mixture was filtered and washed repeatedly with DMF, MC, MC / MeOH, MeOH to give a resin (206.8 mg) as a pale yellow solid represented by the formula (3-3). The obtained resin (3-3) was added to a dichloromethane (5 mL) solution, shaken and trifluoroacetic acid (TFA, 1 mL) was added, followed by shaking at room temperature for 4 hours. After the reaction was completed, the reaction mixture was filtered, the filtrate was washed repeatedly with CH ₂ Cl ₂ and MeOH, and the filtrates were combined and concentrated. Ethyl acetate (3 ml) was added to the concentrated mixture, which was then filtered through SAX resin (strong anion exchange resin) and washed repeatedly with ethyl acetate to remove residual trifluoroacetic acid. The filtrate was concentrated under reduced pressure, and then purified by column chromatography on silica gel under a mixed solvent of hexane / ethyl acetate (4/1, v / v ) to give a pale yellow oil (14.8 mg, Resin 1; yield = 56% from loading capacity of 0.55 mmol / g) was obtained.

M / S 265.36

II-4) Hydroxy Butoxy Addition and Desorption of N-Methyl Substituted Olefin Resin (2a)

N-methyl-substituted olefin resin (200.00 mg, 0.11 mmol) represented by the formula (2a) was added to a mixed solution of dichloromethane (3 mL) and butanol (3 mL), and shaken for 30 minutes, followed by meta-chloroperbenzoic acid ( m- CPBA). ) (81 mg, 0.55 mmol) was added at room temperature, followed by shaking at 35 for 15 hours. After completion of the reaction, the reaction mixture was filtered and washed repeatedly with DMF, MC, MC / MeOH, MeOH to give a resin (208.3 mg) as a pale yellow solid represented by the formula (3-4). The obtained resin (3-4) was added to a dichloromethane (5 mL) solution, shaken and trifluoroacetic acid (TFA) (1 mL) was added, followed by shaking at room temperature for 4 hours. After the reaction was completed, the reaction mixture was filtered, the filtrate was washed repeatedly with CH ₂ Cl ₂ and MeOH, and the filtrates were combined and concentrated. Ethyl acetate (3 ml) was added to the concentrated mixture, which was then filtered through SAX resin (strong anion exchange resin) and washed repeatedly with ethyl acetate to remove residual trifluoroacetic acid. The filtrate was concentrated under reduced pressure, and then purified by column chromatography on silica gel under a mixed solvent of hexane / ethyl acetate (4/1, v / v ) to give a pale yellow oil (19.1 mg, represented by the compound number (4-4), Resin 1; yield = 62% from loading capacity of 0.55 mmol / g) was obtained.

¹ H NMR (200 MHz, CDCl ₃ ) δ (PPM) 6.70 to 6.50 (m, 3H), 4.35 (d, 1H, J = 7.2 Hz), 3.80 (d, 1H, J = 7.2 Hz) 3.67 (t, 2H, J = 6.2 Hz), 2.81 (s, 3H), 1.67-1.53 (m, 2H), 1.49-1.30 (m, 2H), 1.42 (s, 3H), 1.25 (s, 3H), 0.94 (t , 3H, J = 7.2 Hz); M / S 279.38

2,2-dimethyl-3-hydroxy-4-alkoxy-6-alkyl amino benzopyran derivatives synthesized by the solid-phase equilibrium synthesis method in the same manner as in the above embodiment are shown in Table 1 below.

The precursors used to perform the combination of each substituent to complete the library of Table 1 are as follows. For example, R ¹ precursors include CH ₃ I, C ₂ H ₅ I, t BuI, 4-BuI, 4-FC ₄ H ₉ I, 4-Me-C ₄ H ₉ I, 2-CH ₃ -C ₄ H ₉ I, 3-Cl-C ₄ H ₉ I, 4- t Bu-C ₄ H ₉ I, 3-FC ₄ H ₉ I was used, and R ² precursors are CH ₃ OH, C ₂ H ₅ OH, i PrOH, t BuOH, C ₄ H ₉ OH, 2-Ph-EtOH, 2-cyclohexyl-EtOH were used.

.

The following are some examples of formulation methods containing the compound according to the present invention as an active ingredient, but the present invention is not limited thereto.

Formulation 1 : tablet (direct pressure)

After sifting 5.0 mg of 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 form a tablet.

Formulation 2 : Tablet (Wet Granulation)

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 into tablets.

Formulation 3 : Powders and Capsules

5.0 mg of the active ingredient was sieved, followed by mixing with 14.8 mg of lactose, 10.0 mg of polyvinyl pyrrolidone, and 0.2 mg of magnesium stearate. No. solid the mixture using a suitable device. Filled in 5 gelatin capsules.

Formulation 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.

In addition, to investigate the effect of inhibiting lipid peroxidation induced by iron of the compounds according to the present invention, the following experiment was performed.

Experimental Example: Lipid Peroxidation Inhibitory Effect

Rat brains were treated with Krebs buffer (15 mM HEPES, 10 mM glucose, 140 mM NaCl, 3.6 mM KCl, 1.5 mM CaCl ₂ , 1.4 mM KH ₂ PO ₄ , 0.7 mM MgCl ₂ , _P H 7.4) After homogenization, the mixture was centrifuged at 12,000 rpm for 10 minutes, and the supernatant of the brain homogenate was used as a raw material of lipid.

FeCl ₂ was added to the brain homogenate to a final concentration of 400 μM and left at 37 ° C. for 30 minutes to promote oxidation. At this time, 12.5 μM was added to the test material, and only dimethyl sulfoxide (DMSO) was added as a solvent.

The addition of iron to brain homogenates promotes oxidation, increasing the amount of malondialdehyde (MDA), a lipid peroxidation product, and thus the degree of lipid peroxidation was determined by MDA assay. MDA quantitation is typically used to measure absorbance at 530 nm by reacting a sample with 2-thiobarbituric acid (TBA), but is not suitable for processing large samples because of the boiling step. Therefore, in the present invention, N-methyl-2-phenylindole, which is a coloring reagent, was used instead of TBA. In this case, one molecule of MDA and two molecules of N-methyl-2-phenylindole react to form a chromosome, which exhibits a maximum absorbance at 586 nm and does not require boiling (Bioxytech ™ LPO-586 Kit). Absorbance measurements). The lipid peroxidation inhibitory effect of the test substance was calculated as a percentage of the amount of MDA decrease based on the amount of MDA of the control group. For test substances with an inhibitory effect of more than 80%, the concentration-lipid peroxidation effect response curve was calculated by calculating the rate of decrease of MDA amount based on the MDA amount of the control group at various concentrations, and the least linear regression analysis was performed. Through the calculation of IC ₅₀ , a 50% inhibitory concentration by drug administration, the results are shown in Table 2 below.

Table 2 also shows representative antioxidants currently on the market such as Promethazine, Trolox, Probucol and N-propyl gallate. In the same manner as above, the 50% inhibitory concentration (IC ₅₀ ) was calculated and used as a medicament.

As can be seen in Table 2, the compounds of the present invention have the activity of inhibiting lipid peroxidation by iron. In particular, compounds 4-111 and 4-115 show an inhibitory rate of 1.0 μM or less, respectively, and thus, the effect of inhibiting lipid peroxidation by iron is very strong.

As described above, in general, the reaction of the multi-step process is performed in a solution phase, and after several reactions, a treatment process and a purification process must be performed, whereas 2,2-dimethyl-3-hydroxy using the solid phase equilibrium synthesis method according to the present invention. The method for preparing a -6-alkyl amino benzopyran derivative can construct a large amount of libraries in a short time by greatly reducing post-reaction treatment and purification. In particular, in carrying out the reaction according to the present invention, N-alkyl-substituted carbamate resin production reaction represented by formula (2) in the first step, and 2,2-dimethyl- represented by formula (3) in the second step Formation reaction of 3-hydroxy-4-alkoxy-6-alkyl amino benzopyran resin, in the third stage reaction, using the 25% TFA / CH ₂ Cl ₂ solution, the target compound represented by the formula (4) 2,2 The preparation of -dimethyl-3-hydroxy-4-alkoxy-6-alkyl amino benzopyran derivatives was automated using a solid-phase equilibrium method to post-reaction treatment and purification.

Therefore, the present invention has established a technique for constructing a 3-hydroxy benzopyran library using a solid-phase equilibrium synthesis method, thereby increasing the applicability of combinatorial chemical synthesis technology, which promotes lipid peroxidation and accumulates oxides in neurons. Exploration and structure and function of lead compounds with new structures useful for the development of preventive or therapeutic agents for inflammatory diseases such as arthritis, myocardial infarction, acute tissue damage, as well as cerebral nervous system diseases such as stroke and dementia. It's easier to optimize.

Claims (3)

A 2,2-dimethyl-3-hydroxy-4-alkoxy-6-alkylamino benzopyran derivative characterized by being represented by the following formula (4a): [Formula 4a]

In the formula (4a), R ^1a represents an allyl group, benzyl group or substituted benzyl group, naphthylmethyl group, or phenethyl group; R ² represents a C ₁ to C ₁₀ alkyl group, benzyl group or substituted benzyl group, or — (CH ₂ ) _n -phenyl group, where n is an integer of 2 or 3; In this case, the substituted benzyl group represents a benzyl group in which 1 to 4 substituents selected from a halogen atom, a nitro group, a C _{1 to} C ₁₀ alkyl group, a C _{1 to} C ₁₀ alkoxy group, and a C _{1 to} C ₁₀ haloalkyl group are substituted. In the conditions using the reaction solvent of dimethyl sulfoxide or tetrahydrofuran, the base of lithium t-butoxide, and the electrophile represented by R <^1> -X (where X is halogen), it is represented by following General formula (1) Synthesizing an N-alkyl substituted carbamate resin represented by the following formula (2) by selectively introducing an alkyl substituent to the nitrogen atom of benzopyran linked with a carbamate linker; Under conditions using a dichloromethane solvent, meta-chloroperbenzoic acid ( m- CPBA) and an alcohol were added to a compound represented by the following formula (2) to simultaneously perform an epoxidation reaction and an alkoxy addition reaction to the following formula (3). Synthesizing the resin in the form of the indicated 2,2-dimethyl-3-hydroxy-6-alkyl amino benzopyran; And 2,2-dimethyl-3-hydroxy-4-alkoxy represented by the following formula (4) by desorption using a dichloromethane solution containing trifluoroacetic acid (TFA) Method for producing a -6-alkyl amino benzopyran derivative comprising the step of:

In the above scheme, R ¹ is C ₁ ~C ₁₀ alkyl group, an allyl group, a benzyl group or substituted benzyl group, a naphthylmethyl group, or the pen represents an ethyl group; R ² represents a C ₁ to C ₁₀ alkyl group, benzyl group or substituted benzyl group, or — (CH ₂ ) _n -phenyl group, where n is an integer of 2 or 3; Wherein the substituted benzyl group represents a benzyl group in which 1 to 4 substituents are selected from a halogen atom, a nitro group, a C _{1 to} C ₁₀ alkyl group, a C _{1 to} C ₁₀ alkoxy group and a C _{1 to} C ₁₀ haloalkyl group; Ⓟ represents a solid support in the form of a high molecular polymer selected from polystyrene-divinylbenzene, methacrylic acid-dimethylacrylamide and hydroxy methacrylic acid A 2,2-dimethyl-3-hydroxy-4-alkoxy-6-alkylamino benzopyran derivative represented by the following formula (4) contains a lipid peroxidation inhibitory activity:

In the formula (4), R ¹ is C ₁ ~C ₁₀ alkyl group, an allyl group, a benzyl group or substituted benzyl group, a naphthylmethyl group, or the pen represents an ethyl group; R ² represents a C ₁ to C ₁₀ alkyl group, benzyl group or substituted benzyl group, or — (CH ₂ ) _n -phenyl group, where n is an integer of 2 or 3; In this case, the substituted benzyl group represents a benzyl group in which 1 to 4 substituents selected from a halogen atom, a nitro group, a C _{1 to} C ₁₀ alkyl group, a C _{1 to} C ₁₀ alkoxy group, and a C _{1 to} C ₁₀ haloalkyl group are substituted.