KR100746939B1 - N-[1'-substituted sulfonamide-spiro(2h-1-benzopyrane-2,4-piperidin) -6-yl]substituted amine derivatives, process for preparation and use thereof - Google Patents

Abstract

N-[1'-substituted sulfonamide-spiro(2H-1-benzopyrane-2,4-piperidin)-6-yl]substituted amine derivatives, a process for preparation of the same compounds and use thereof are provided to inhibit 5-lipoxygenase and improve simplicity and yield of preparation, so that the compounds are useful for prevention and treatment of diseases caused by activation of leukotriens. The N-[1'-substituted sulfonamide-spiro(2H-1-benzopyrane-2,4-piperidin)-6-yl]substituted amine derivatives represented by the formula(1) and pharmaceutically acceptable salts thereof are provided, wherein R^1 is C1-C10 alkyl group, benzyl or substituted benzyl group, 2-pyrimidylmethyl group, thiophene group, 2-methylthiophenemethyl group, 5-methyl-2-thiophenemethyl group, indolylmethyl group, benzo dioxolanylmethyl group, naphthalenylmethyl group or furanylmethyl group; and R^2 is C1-C10 alkyl group, phenyl or substituted phenyl group, or hetero atom-containing 5- to 7-membered hetero ring and corresponding sulfonyl group.

Description

N- [1′-substituted sulfonamide-spiro (2H-1-benzopyran-2,4-piperidin) -6-yl] substituted amine derivatives having inflammatory inhibitory activity, pharmaceutically acceptable salts thereof, Production method thereof and use thereof {N- [1'-SUBSTITUTED SULFONAMIDE-SPIRO (2H-1-BENZOPYRANE-2,4-PIPERIDIN) -6-YL] SUBSTITUTED AMINE DERIVATIVES, PROCESS FOR PREPARATION AND USE THEREOF}

The present invention has a 1'-substituted sulfonamide-spirobenzopyran as a basic skeleton, N- [1'-substituted sulfonamide-spiro (2H-1-benzopyran-2, 4-piperidin) -6-yl] substituted amine derivatives, pharmaceutically acceptable salts, methods of synthesizing the novel compounds described above with high efficiency, and novel uses thereof.

5-Lipoxygenase (hereinafter referred to as "5-LO"), an inflammation mediator, is an enzyme that is involved in the metabolism of arachidonic acid. It is 5-HPETE (5-hydroperoxyeicosatetranoic acid from arachidonic acid). ) Works with the 5-LO enzyme to produce leukotriene. Among the leukotrienes produced, leukotriene (LTB4), a strong chemoattractant, is known as a major cause of various diseases such as chronic inflammation, rheumatoid arthritis, allergy, asthma, psoriasis. In other words, an increase in leukotriene in cells activates inflammatory cells, causing inflammatory diseases such as chronic inflammation and rheumatism, as well as injuring tissues or infecting organs with endogenous toxins. It is known to cause damage.

Therefore, in recent years, by developing a 5-lipoxygenase (5-LO) inhibitor that can inhibit the activation of inflammatory cells by the increase of intracellular leukotriene to prevent tissue damage, various diseases caused by inflammation Is trying to prevent or treat it.

On the other hand, combinatorial chemical synthesis is a new synthesis technology for new material development, whereas the conventional organic synthesis technology synthesizes a single compound in one reaction. Compounds are being spotlighted as high-efficiency chemical synthesis that can simultaneously synthesize or automate multiple stages of synthesis. Therefore, the introduction of such combinatorial chemical synthesis techniques facilitates the search for new structures of biological hit compounds and lead compounds and optimization of their structure and activity.

In particular, the efficient construction of large-scale and concentrated libraries of combinatorial chemical synthesis techniques for organic low-molecular compounds that allow for the introduction of various substituents into a molecule but does not deviate significantly from the scope of Lipinsky's rule of 5 This is important in order to secure molecular diversity that is useful for the exploration of leading substances.

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.

However, in spite of the advantages of the combined chemical synthesis technology, since most of the chemical reaction proceeds on the solid support, the reaction reagent is used in excess, causing unwanted side reactions, and the solvent to be used depending on the physical properties of the solid support used. Because of the limited range of chemical reactions that can be selected, it could not be easily applied to the field of organic synthesis.

More specifically, Merrifield resin and Wang resin are widely used as conventional solid supports used in combinatorial chemistry. However, since the solid support has extremely low swelling effect in polar solvents such as alcohol and water, there are many limitations in the selection of a solvent required for the reaction.

Therefore, in order to synthesize various derivatives using solid phase chemical reactions, it is necessary to select the desired solid support and linker, to search for reaction reagents and reaction conditions, and to change various chemical structures and physical properties of the target compound. The choice of substituents is an important factor.

In the field of new material development, in the development of compounds having pharmacological effects such as neurological diseases, hypertension drugs, and diabetes drugs, natural products and compounds based on 1'-substituted sulfonamide-spiroben Zopyran based on It is revealed that the compound possesses pharmacological efficacy, and its application has been progressed. However, the development of a compound based on 1'-substituted sulfonamide-spirobenzopyran as a therapeutic agent for inflammatory diseases has not been reported.

Accordingly, the present inventors have tried to synthesize biologically effective compounds and lead compounds useful in treating inflammatory diseases with derivatives having 1'-substituted sulfonamide-spirobenzopyran as a basic skeleton. N- [1'-substituted sulfonylamide-spy is economically and efficiently constructed by constructing 1'-substituted sulfonamide-spirobenzopyran library using solid phase equilibrium synthesis method in combinatorial chemical synthesis technology. Efficiently synthesizing rho (2H-1-benzopyran-2,4-piperidin) -6-yl] substituted amine derivatives, and according to the 5-LO inhibitory activity of the compound, By providing it, this invention was completed.

An object of the present invention is N- [1'-substituted sulfonamide-spiro (2H-1-benzopyran), which is a novel compound having 1'-substituted sulfonamide-spirobenzopyran as a basic skeleton and having inflammation inhibitory activity. -2,4-piperidin) -6-yl] substituted amine derivatives and pharmaceutically acceptable salts.

It is another object of the present invention to substitute the N- [1'-substituted sulfonamide-spiro (2H-1-benzopyran-2,4-piperidin) -6-yl] substituted by solid-state equilibrium synthesis of combinatorial chemistry It is to provide a method for producing an amine derivative.

Another object of the present invention is to provide a novel use of the N- [1′-substituted sulfonamide-spiro (2H-1-benzopyran-2,4-piperidin) -6-yl] substituted amine derivative It is.

The present invention provides N- [1'-substituted sulfonamide-spiro (2H-1-benzopyran-2,4-piperidin) -6-yl] substituted amine derivative represented by the following formula (1) and pharmaceutically acceptable Provide possible salts.

(Wherein, R ¹ is C ₁ ~C ₁₀ alkyl group, a benzyl or substituted benzyl group, a phenethyl group, 2-pyrimidyl group, thiophenyl group, a 2-thiophene group, 5-methyl-2-thiophene Methyl group, 3-thiophenmethyl group, indolylmethyl group, benzodioxoranylmethyl group, naphthalenylmethyl group, furanylmethyl group, or 2-furanmethylmethyl group; R ² is C ₁ -C ₁₀ alkyl group, phenyl or substituted A phenyl group or a 5 to 7 membered heterocycle including a hetero atom selected from oxygen and a sulfur atom, and a sulfonyl group corresponding thereto, wherein the benzyl group, phenyl group or heterocyclic group is C ₁ -C ₆ alkyl group, C _{1- 1} to 4 substituents selected from a C ₆ haloalkyl group, a halogen, a nitro group, a cyano group, and a C _{1 to} C ₆ alkoxy group may be substituted.)

N- [1′-Substituted sulfonamide-spiro (2H-1-benzopyran-2,4-piperidin) -6-yl] substituted amine derivatives of the invention and pharmaceutically acceptable salts are Foxy genease inhibitory activity.

The present invention provides a process for the preparation of N- [1′-substituted sulfonamide-spiro (2H-1-benzopyran-2,4-piperidin) -6-yl] substituted amine derivatives.

More specifically, the N- [1′-substituted sulfonamide-spiro (2H-1-benzopyran-2,4-piperidin) -6-yl] substituted amine derivative according to the present invention is combined chemical synthesis The reaction to be produced is shown in Scheme 1 below. The manufacturing method of the present invention,

1) N- [1'-Fmoc-protected amide-spiro (2H-1-benzopyran-2,4) represented by the following formula (3) to a solid support linked to a methoxy benzaldehyde resin represented by the following formula (2) by a linker -Piperidin) -6-yl] amine was introduced to carry out the reaction of N- [1'-Fmoc protecting amide-spiro (2H-1-benzopyran-2,4-piperi) Din) -6-yl] amine resin;

2) Nitrogen atom of 6-amino group of N- [1'-Fmoc protectingamide-spiro (2H-1-benzopyran-2,4-piperidin) -6-yl] amine resin represented by the following general formula (4) A reaction of the R ¹ substituent is optionally carried out to N- [1′-Fmoc protecting amide-spiro (2H-1-benzopyran-2,4-piperidine) -6- A second step of synthesizing the general substituted amine resin;

3) N- [1′-Fmoc protecting amide-spiro (2H-1-benzopyran-2,4-piperidin) -6-yl] substituted amine resin compound represented by the following formula (5) N- [1'-amide-spiro (2H-1-benzopyran-2,4-piperidine) represented by the following Chemical Formula 6 by desorption using a dichloromethane solution or an organic solvent containing an organic base A third step of synthesizing) -6-yl] substituted amine resin;

4) Nitrogen of the 2'-aminomethyl group of N- [1'-amide-spiro (2H-1-benzopyran-2,4-piperidin) -6-yl] substituted amine resin represented by following General formula (6). A reaction of introducing an R ² substituent to an atom is carried out selectively, whereby N- [1′-substituted sulfonylamide-spiro (2H-1-benzopyran-2,4-piperidine) -6 represented by the following formula (7): A fourth step of synthesizing a -yl] substituted amine resin; And

5) Trifluoroacetic acid of N- [1'-substituted sulfonylamide-spiro (2H-1-benzopyran-2,4-piperidin) -6-yl] substituted amine resin compound represented by the following formula (7): N- [1'-substituted sulfonylamide-spiro (2H-1-benzopyran-2) represented by the following formula (1) by desorption using a dichloromethane solution containing (TFA) or an organic solvent containing an organic acid. And a fifth step of synthesizing, 4-piperidin) -6-yl] substituted amine derivative.

(In the above, R ¹ and R ² are as defined above; ⓟ is a solid support selected from the group consisting of polystyrene-divinylbenzene, methacrylic acid-dimethylacrylamide and hydroxyl methacrylic acid.)

The preparation method of the present invention is carried out using a solid phase equilibrium synthesis method in a combination chemical synthesis technique, a method capable of automating a continuous multi-step reaction and reaction process, the present invention is a methoxy on a solid support represented by the formula (4) N- [1′-Fmocprotectamide-spiro (2H-1-benzopyran-2,4-piperidin) -6-yl] amine resin, wherein the benzaldehyde resin is linked by a linker and is represented by Formula 6 Two N-substitution reactions were carried out in equilibrium using N- [1'-amide-spiro (2H-1-benzopyran-2,4-piperidin) -6-yl] substituted amine resin As a result, various N- [1′-substituted sulfonylamide-spiro (2H-1-benzopyran-2,4-piperidine)-in a short time since several reactions and purification processes can be performed simultaneously. 6-yl] substituted amine derivatives can be synthesized.

Accordingly, the selection of the solid support and linker, the reaction reagent and the reaction conditions, and the selection of the substituent which can variously change the chemical structure and physical properties of the target compound will be described as 1'-substituted sulfonamide-spyro. It will of course be understood that it is an optimal condition for building the benzopyran library.

Specifically, the range of selection of the reaction process, solvent composition and reaction conditions in carrying out the production method according to the present invention will be described in detail.

The preferred solvent used in the production method of the present invention is an organic solvent with excellent swelling effect, and uses Wang resin or Merrifield resin.

The first stage reaction is N- [1′-Fmocprotectamide-spiro (2H-1-benzopyran-2,4-piperidin) -6-yl represented by Formula 3 on a solid support represented by Formula 2 ] To introduce N- [1'-Fmoc protecting amide-spiro (2H-1-benzopyran-2,4-piperidin) -6-yl] amine resin represented by formula (4). As a step of synthesizing, preferred solvent is dimethylformamide (DMF) containing 1% acetic acid in order to carry out the reductive amination reaction. The reducing agent NaBH (OAc) used in the reaction is preferably used in an amount of 3 equivalents or less, and more preferably in an amount of 2 equivalents or less.

The second stage reaction was carried out of the 6-amino group of N- [1′-Fmocprotectamide-spiro (2H-1-benzopyran-2,4-piperidin) -6-yl] amine resin represented by the formula (4). N- [1'-Fmoc-protected amide-spiro (2H-1-benzopyran-2,4-piperidine) -6 represented by the formula (5) by selectively introducing a R ¹ substituent to a nitrogen atom Synthesizing a -yl] substituted amine resin, dichloromethane (MC) or tetrahydrofuran (THF) is used as a preferred solvent in the introduction of the R ¹ substituent.

At this time, it is preferable to use the base and the R ¹ substituent in 3 equivalents or less in the reaction, preferably, it is advantageous in terms of economics to use in the range of about 2 equivalents. At this time, the base used is an organic base having a large steric hindrance such as diisopropylethylamine (DIPEA), and the like, and the base is used in an amount of 3 equivalents or less, more preferably 2 equivalents or less, which is excellent in economic efficiency.

The third stage reaction is triethyl N- [1′-Fmocprotectamide-spiro (2H-1-benzopyran-2,4-piperidin) -6-yl] substituted amine resin compound represented by the formula (5). N- [1'-amide-spiro (2H-1-benzopyran-2,4-piperi) represented by the formula (6) by desorption using a dichloromethane solution containing an amine or an organic solvent containing an organic base. Din) -6-yl] substituted amine resin.

In the desorption reaction of the 2'-Fmoc protecting group, dichloromethane (MC) or tetrahydrofuran (THF) is used as a solvent, and triethylamine base is used in the range of about 3 equivalents or less, more preferably about 2 equivalents. do.

The fourth stage reaction is a 2'-aminomethyl group of N- [1'-amide-spiro (2H-1-benzopyran-2,4-piperidin) -6-yl] substituted amine resin represented by the formula (6). N- [1′-substituted sulfonylamide-spiro (2H-1-benzopyran-2,4-piperidine)-represented by the formula (7) by selectively introducing R ² substituent into the nitrogen atom of 6-yl] substituted amine resin is synthesized.

In the above reaction, in the introduction reaction of a selective R ² substituted sulfonyl group to the nitrogen atom of spiropyridine, position 2 of benzopyran, it is used as a dimethylformamide solvent, and an organic base such as triethylamine and R ² substitution It is preferable to use about 3 equivalents of each of the polyvinyl precursors, and it is preferable to use it in the range of about 2 equivalents preferably.

The fifth step reaction was carried out using N- [1′-substituted sulfonylamide-spiro (2H-1-benzopyran-2,4-piperidin) -6-yl] substituted amine resin compound represented by the formula (7). N- [1'-substituted sulfonylamide-spiro (2H-1-benzopyran-2) represented by Chemical Formula 1 by desorption using a dichloromethane solution containing roacetic acid (TFA) or an organic solvent containing an organic acid. It is a step of synthesizing the target compound of the, 4-piperidin) -6-yl] substituted amine derivative.

The reaction can be synthesized by performing a desorption reaction using a dichloromethane solution containing trifluoroacetic acid (TFA) or an organic solvent containing an organic acid.

Subsequently, N- [1′-substituted sulfonamide-spiro (2H-1-benzopyran-2,4-piperidin) -6-yl] substituted amine derivative represented by Chemical Formula 1 as the target compound is present. In order to confirm the reaction, N- [1′-substituted sulfonamide-spiro (2H-1-benzopyran-2,4-piperidin) -6-yl] substituted in the final step after the reaction The target compound desorbed from the amine resin was purified and separated by flash column chromatography using a multi-column apparatus, and then analyzed by NMR and Mass spectra. The structure was represented by the chemical formulas 2, 4, 5, 6 and 7 Resin can measure the progress of the reaction by measuring the ATR-FTIR.

The present invention provides an N- [1'-substituted sulfonylamide-spiro (2H-1-benzopyran-2,4-piperidin) -6-yl] substituted amine derivative and a pharmaceutically acceptable salt as an active ingredient. Provided is an inflammatory disease treatment.

5-Lipoxygenase (5-LO from N- [1′-Substituted Sulfonylamide-Spiro (2H-1-benzopyran-2,4-piperidin) -6-yl] substitutedamine derivative of the present invention ), The inhibitory activity is confirmed, it can be applied to any inflammatory disease caused by the activity of leukotriene (LTA4, B4, C4, D4). As a specific example, the present invention can be used for the prevention and treatment of various inflammatory diseases such as chronic inflammation, rheumatoid arthritis, colitis, psoriasis, and asthma, which is a kind of inflammatory disease occurring in the airways.

Pharmaceutically acceptable salts in the present invention can be prepared by conventional methods in the art, any one inorganic selected from the group consisting of hydrochloric acid, hydrogen bromide, sulfuric acid, sodium hydrogen sulfate, phosphoric acid and carbonic acid Salts with acids; Salts with any one of organic acids selected from the group consisting of formic acid, acetic acid, oxalic acid, benzoic acid, citric acid, tartaric acid, gluconic acid, gestyic acid, fumaric acid, lactobionic acid, salicylic acid, and acetylsalicylic acid (aspirin) ; It may be reacted with a metal salt or ammonium ion obtained by reaction with an alkali metal ion selected from sodium or potassium to form another form of a pharmaceutically acceptable salt.

In addition, the pharmaceutical composition of the present invention may be a N- [1′-substituted sulfonamide-spiro (2H-1-benzopyran-2,4-piperidin) -6-yl] substituted amine derivative or a pharmaceutically acceptable To these salts, conventional non-toxic pharmaceutically acceptable carriers, adjuvant and excipients are added, and for oral administration of conventional agents in the pharmaceutical field, for example, tablets, capsules, troches, solutions, suspensions and the like. It may be formulated into a formulation or a parenteral formulation.

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.

Hereinafter, the present invention will be described in detail by way of examples.

The following examples are merely illustrative of the present invention, but the scope of the present invention is not limited to the following examples.

Example 1 Synthesis of N- [1′-Substituted Sulfonamide-Spyro (2H-1-benzopyran-2,4-piperidin) -6-yl] substituted amine derivative (Formula 1) 1

First Step: Synthesis of N- [1′-ethylcarbamateprotectedamino-spiro (2H-1-benzopyran-2,4-piperidin) -6-yl] amine resin (Formula 4)

Methoxy Benzaldehyde Resin (AMEBA) represented by the formula (2) (1.6 mmol / g, 10 g, 16.0 mmol) was added to 100 ml of dimethylformamide containing 1% acetic acid, and N- [1'- 9.28 g (32.0 mmol) of ethylcarbamateprotectamide-spiro (2H-1-benzopyran-2,4-piperidin) -6-yl] amine with sodium triacetoxyboron hydride (NaBH (OAc) ₃ 6.78g (32.0mmol) was added continuously, and the mixture was shaken at room temperature for 15 hours.

After completion of the reaction, the reaction mixture was filtered and washed repeatedly with dimethylformamide, dichloromethane, dichloromethane / methanol and methanol to obtain 12.1 g of a solid resin represented by Chemical Formula 3 (ATR-FTIR; carbamate: 1694 cm ^-1 ) .

0.200 g (0.32 mmol) of the obtained resin was added to a 5 mL solution of dichloromethane, shaken, 1 mL of trifluoroacetic acid was added thereto, followed by shaking at room temperature for 4 hours.

After the reaction was completed, the reaction mixture was filtered, the filtrate was repeatedly washed with CH ₂ Cl ₂ and methanol, and the filtrates were combined and concentrated. After adding 3 ml of ethyl acetate to the concentrated mixture, the resultant was filtered through an anion exchange resin (SAX 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 (3/1, v / v) to give 67.3 mg (73%; 1.2 mmol / g of loading capacity of the resin was obtained.

Second Step: N-Benzylation of N- [1′-ethylcarbamateprotectedamino-spiro (2H-1-benzopyran-2,4-piperidin) -6-yl] amine resin (Formula 4) reaction

0.200 g (0.24 mmol) of benzopyrene resin represented by Chemical Formula 4 was added to 3 ml of dimethylformamide, shaken at room temperature for 10 minutes, and 0.14 ml (0.72 mmol) of benzyl bromide and 0.12 ml (0.72 mmol) of diisopropylamine were added thereto. Shake for 15 hours at room temperature and mix. After completion of the reaction, the reaction mixture was filtered and washed repeatedly with dimethylformamide, dichloromethane, dichloromethane / methanol and methanol to obtain a resin which is a pale brown solid of Formula 5-1.

Third Step: N- [1′-Ethyl Carbamate-Protected Amino-Spyro (2H-1-benzopyran-2,4-piperidin) -6-yl] amine Carbamate- of Resin (Formula 4)- Deprotection reaction

The obtained resin of Formula 5-1 was added to a 5 ml mixed solution of butanol / 1,4-dioxane containing potassium hydroxide, and shaken at 90 ° C for 3 hours. After the reaction was completed, the reaction mixture was filtered and washed repeatedly with dimethylformamide, dichloromethane, dichloromethane / methanol and methanol to obtain a resin which is a pale brown solid of Formula 6-1 (ATR-FTIR; disappearing carbamate peaks: 1694 cm-) ^{. 1} ).

Fourth Step: N-sulfonation with N- [1′-amide-spiro (2H-1-benzopyran-2,4-piperidin) -6-yl] benzylamine resin (Formula 6-1) reaction

0.200 g (0.24 mmol) of benzopyrene resin represented by Chemical Formula 6-1 was added to 3 ml of dimethylformamide, and stirred at room temperature for 10 minutes, and 0.148 mg (0.72 mmol) of benzenesulfonyl chloride (4-MeOPhSO ₂ Cl) and diisopropyl 0.12 mL (0.72 mmol) of amine was added thereto, followed by mixing for 15 hours at room temperature. After completion of the reaction, the reaction mixture was filtered and washed repeatedly with dimethylformamide, dichloromethane, dichloromethane / methanol and methanol to obtain a resin which is a pale brown solid of Formula 7-1.

Fifth Step: Solid Support Desorption Reaction of Formula 7-1

0.200 g (0.24 mmol) of the obtained resin of Chemical Formula 7-1 was added to a 5 ml solution of dichloromethane, shaken, and 1 ml of trifluoroacetic acid was added thereto, followed by shaking at room temperature for 4 hours. After the reaction was completed, the reaction mixture was filtered, the filtrate was repeatedly washed with CH ₂ Cl ₂ and MeOH, and the filtrates were combined and concentrated. 3 ml of ethyl acetate 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 (3/1, v / v) to give 81 mg (71%; resin) of the pale brown oil represented by the formula (1-1). 1.2 mmol / g loading capacity of 4) was obtained.

Examples 2 to 96 Synthesis of N- [1′-Substituted Sulfonamide-Spyro (2H-1-benzopyran-2,4-piperidin) -6-yl] substituted amine derivatives 2 to 96

In the compound represented by Formula 1, R ¹ and R ² N- [1′-Substituted sulfonamide-spiro (2H-1-benzopyran-2,4) was carried out in the same solid phase equilibrium method as in Example 1, except that it was carried out as described in Table 1 below . -Piperidin) -6-yl] substituted amine derivative was synthesized.

Formula 1

(In the above, R ¹ And R ² is as defined above.)

< Experimental Example  1> bioassay test: 5- Lipoxygenase assay

For the N- [1′-substituted sulfonamide-spiro (2H-1-benzopyran-2,4-piperidin) -6-yl] substituted amine derivative prepared in the above example, 5-Lipoxygenase enzyme was prepared. The bioassay test used was performed as follows.

1. Assay by FOX Reagent

After 7 μg of insect cell lysates expressing 5-Lipoxygenase (hereinafter referred to as “5-LO”) was added to the test compound (final concentration of 1 μM) and reacted at room temperature for 3 minutes, the enzyme substrate arachidonic acid 40 μM was added and allowed to react at room temperature for 4 minutes. 100 μl of FOX reagent (25 mM sulfuric acid, 100 μM xylenol orange, 100 μM FeSO ₄ , methanol: water = 9: 1) was added thereto, and absorbance was measured at 575 nm after 5 minutes.

2. Spectroscopic Assay (234 nm)

After 7 μl of 5-LO expressing insect cell lysates, test compound (final concentration: 1 μM) was added and reacted at room temperature for 3 minutes, followed by reaction buffer (50 mM Tris buffer, pH 7.4, 0.4 mM CaCl ₂ , 24 μg / mL phosphadidylcholine, 40 μM arachidonic acid) was added and the change in absorbance at 234 nm at room temperature was measured for 4 minutes.

Experimental Example 2 Bioassay: LTB4 cell-based assay

For the N- [1′-substituted sulfonamide-spiro (2H-1-benzopyran-2,4-piperidin) -6-yl] substituted amine derivative prepared in the above Example, LTB4 production using cells Inhibition measurement experiments were performed as follows.

The rat basophilic leukemia (RBL-1) cells were seeded in a 24 well so that the cell number was 7.5 × 10 ⁵ , and then reacted for 2 hours to attach the cells to the bottom. After 2 hours, 10 μM (final) A23187 was treated for 10 minutes, and the test compound (final concentration 10 μM) was added to react for 10 minutes. Thereafter, the supernatant was collected by centrifugation at 1500 g for 20 minutes, and the amount of LTB4 was measured using an ELISA method.

Experimental Example 3 Bioassay Test: In vivo  assay; Mouse ear edema model

For the N- [1′-substituted sulfonamide-spiro (2H-1-benzopyran-2,4-piperidin) -6-yl] substituted amine derivative prepared in the above example, the organism using the animal model Assay experiments were performed as follows.

Inflammation was induced for 1 hour by treatment with 2 mg arachidonic acid dissolved in 20 μL of acetone inside the right ear of 6-week old mice.

The difference between the right ear and untreated left ear after 1 hour of arachidonic acid treatment was measured using a microgauge. In addition, the activity of the mieroperoxidase (MPO) enzyme was measured to determine the amount of necrophilic leukocytes (neurophil) infiltrated into the tissue as an indicator of early inflammation. The mieroperoxidase (MPO) is mainly contained in neurogenic leukocytes (neurophil), since it is expressed first in acute inflammation, it can be used as an indicator of inflammatory judgment. In other words, a part of the arachidonic acid-treated ears was obtained by crushing the tissue in 50 mM phosphate buffer (pH 6.0) containing 0.5% hexadecyltrimethylammonium bromide (HTAB), and then centrifuging to measure the myeloperoxidase (MPO) activity of the supernatant. .

To measure the in vivo effect of the test compound, the compound was dissolved in 0.5% methyl cellulose (10 mL / kg), orally administered 1 hour before arachidonic acid treatment, and treated with arachidonic acid, and 1 hour after ear thickness and MPO activity. The in vivo efficacy of the test compound was evaluated by comparing with a control treated with only arachidonic acid without treating the compound. At this time, the thickness measurement is to measure the degree of edema observed during inflammation.

The results for the bioassay test are shown in Table 2 .

As can be seen in Table 2, N- [1'-substituted sulfonamide-spiro (2H-1-benzopyran-2,4-piperidin) -6-yl] substituted in the above Example Amine derivatives confirmed inhibitory activity against the 5-LO enzyme.

In particular, the N- [1′-substituted sulfonamide-spiro (2H-1-benzopyran-2,4-piperidin) -6-yl] substituted amine derivatives of the present invention may be prepared by the enzyme, cell and animal experiments. In the results, it was confirmed that a strong effect that can compete with the reference drug, Zilutone.

N- [1'-substituted sulfonamide-spiro (2H-1-benzopyran-2,4-piperidin) -6-yl] substituted amine derivative of the present invention is a hydroxy urea form of Zilutone Although the chemical structure is different, by confirming 5-LO inhibitory activity, it can be usefully used in the development of drugs such as new chronic inflammation, rheumatoid arthritis, colitis, asthma, psoriasis.

The following formulation example contains N- [1′-substituted sulfonamide-spiro (2H-1-benzopyran-2,4-piperidin) -6-yl] substituted amine derivative according to the present invention as an active ingredient. It is merely illustrative of the formulation, but is not limited thereto.

Preparation Example 1 Preparation of Tablet (Direct Press Method)

As an active ingredient, 5.0 mg of the derivative compound of the present invention was sieved, and then 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.

Preparation Example 2 Preparation of Tablet (Wet Granulation)

As an active ingredient, 5.0 mg of the derivative compound of the present invention was sieved, followed by mixing 16.0 mg of lactose and 4.0 mg of starch. 0.3 mg of polysorbate 80 was dissolved in pure water, and then an appropriate amount of the solution was added, followed by atomization. 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.

Preparation Example 3 Preparation of Powder and Capsule

As an active ingredient, 5.0 mg of the derivative compound of the present invention was sieved, and then mixed together 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.

<Example 4> Preparation of Injectables

As an active ingredient, 100 mg of the derivative compound of the present invention was contained, and in addition, 180 mg of mannitol, 26 mg of Na ₂ HPO ₄ .12H ₂ O and 2974 mg of distilled water were used to prepare an injection.

As described above, the present invention

First, while the reaction of the multi-step process is generally carried out in solution and prepared after several reaction treatments and purification processes, N- [1′-substituted sulfonamide-spiro using solid phase synthesis according to the present invention ( The method for preparing 2H-1-benzopyran-2,4-piperidin) -6-yl] substituted amine derivative efficiently simplifies a large amount of drug-like libraries by simplifying the treatment and purification after the reaction. Can be built.

Second, N- [1'-Fmocprotectamide-spiro (2H-1-benzopyran-2,4-piperidin) -6-yl, linked by an AMEBA linker on a solid support represented by Formula 4 of the present invention ] Amine resin and two times using N- [1'-amide-spiro (2H-1-benzopyran-2,4-piperidin) -6-yl] substituted amine resin represented by Formula 6 When the N-substitution reaction is carried out by an equilibrium synthesis method, several N- [1'-substituted sulfonylamide-spiro (2H-1-benzopyran- Extremely useful for the synthesis of 2,4-piperidin) -6-yl] substituted amine derivatives.

Third, the present invention provides a technique for constructing a N- [1′-substituted sulfonamide-spiro (2H-1-benzopyran-2,4-piperidin) -6-yl] substituted amine library using solid phase equilibrium synthesis. By establishing, the applicability of the combinatorial chemical synthesis technology was improved.

Fourth, by confirming the 5-LO inhibitory activity against the target compound of the present invention, inflammatory diseases, rheumatoid arthritis, colitis, asthma, psoriasis, etc. caused by the activity of leukotriene (LTA4, B4, C4, D4) It can be usefully used as a medicament against inflammatory diseases.

Although the present invention has been described in detail only with respect to the embodiments described, it will be apparent to those skilled in the art that various modifications and variations are possible within the technical spirit of the present invention, and such modifications and variations belong to the appended claims. .

Claims (6)

N- [1′-substituted sulfonamide-spiro (2H-1-benzopyran-2,4-piperidin) -6-yl] substituted amine derivative represented by the following formula (1) and a pharmaceutically acceptable salt. Formula 1

(Wherein, R ¹ is C ₁ ~C ₁₀ alkyl group, a benzyl or substituted benzyl group, a phenethyl group, 2-pyrimidyl group, thiophenyl group, a 2-thiophene group, 5-methyl-2-thiophene Methyl group, 3-thiophenmethyl group, indolylmethyl group, benzodioxoranylmethyl group, naphthalenylmethyl group, furanylmethyl group, or 2-furanmethylmethyl group; R ² is C ₁ -C ₁₀ alkyl group, phenyl or substituted A phenyl group or a 5 to 7 membered heterocycle including a hetero atom selected from oxygen and a sulfur atom, and a sulfonyl group corresponding thereto, wherein the benzyl group, phenyl group or heterocyclic group is C ₁ -C ₆ alkyl group, C _{1- 1} to 4 substituents selected from a C ₆ haloalkyl group, a halogen, a nitro group, a cyano group, and a C _{1 to} C ₆ alkoxy group may be substituted.) The derivative and pharmaceutically acceptable salt of claim 1, wherein the derivative and the pharmaceutically acceptable salt have 5-lipoxygenase inhibitory activity. N- [1′-Fmocprotectamide-spiro (2H-1-benzopyran-2, represented by Formula 3) to a solid support connected by a Methoxy Benzaldehyde Resin linker represented by Formula 2, Induction reaction of 4-piperidin) -6-yl] amine was carried out to obtain N- [1'-Fmoc protecting amide-spiro (2H-1-benzopyran-2,4-pi) A first step of synthesizing ferridin) -6-yl] amine resin; Alternatively to the nitrogen atom of the 6-amino group of N- [1′-Fmocprotectedamide-spiro (2H-1-benzopyran-2,4-piperidin) -6-yl] amine resin represented by formula (4) Induction reaction of the R ¹ substituent was carried out to substitute N- [1′-Fmoc protectingamide-spiro (2H-1-benzopyran-2,4-piperidin) -6-yl] represented by the following Chemical Formula 5. A second step of synthesizing the amine resin; N- [1'-Fmoc-protected amide-spiro (2H-1-benzopyran-2,4-piperidin) -6-yl] substituted amine resin compound represented by the formula (5) is dichloro N- [1'-amide-spiro (2H-1-benzopyran-2,4-piperidine) -6 represented by Chemical Formula 6 by desorption using a methane solution or an organic solvent containing an organic base A third step of synthesizing a -yl] substituted amine resin; Selective to nitrogen atom of 2'-aminomethyl group of N- [1'-amide-spiro (2H-1-benzopyran-2,4-piperidin) -6-yl] substituted amine resin represented by the formula (6) Introduced reaction of the R ² substituent, N- [1'-substituted sulfonylamide-spiro (2H-1-benzopyran-2,4-piperidin) -6-yl] represented by the following formula (7) A fourth step of synthesizing a substituted amine resin; And N- [1'-substituted sulfonylamide-spiro (2H-1-benzopyran-2,4-piperidin) -6-yl] substituted amine resin compound represented by the formula (7) is trifluoroacetic acid (TFA) Desorption using a dichloromethane solution or an organic solvent containing an organic acid, N- [1'-substituted sulfonylamide-spiro (2H-1-benzopyran-2,4-py) represented by the following formula (1) A fifth step of synthesizing ferridin) -6-yl] substituted amine derivative; A method for producing the N- [1′-substituted sulfonylamide-spiro (2H-1-benzopyran-2,4-piperidin) -6-yl] substituted amine derivative according to claim 1, comprising: Scheme 1

(In the above, R ¹ and R ² are as defined above; ⓟ represents a solid support selected from the group consisting of polystyrene-divinylbenzene, methacrylic acid-dimethylacrylamide and hydroxyl methacrylic acid.) N- [1′-substituted sulfonylamide-spiro (2H-1-benzopyran-2,4-piperidin) -6-yl] substituted amine derivative of claim 1 and a pharmaceutically acceptable salt thereof as an active ingredient Inflammatory disease treatment characterized in that it contains. The inflammatory disease therapeutic agent according to claim 4, wherein the N- [1'-substituted sulfonylamide-spiro (2H-1-benzopyran-2,4-piperidin) -6-yl] substituted amine derivative of claim 1 is used. The therapeutic agent for inflammatory diseases, characterized in that it is useful for inflammatory diseases caused by the activity of leukotriene by 5-lipoxygenase inhibitory activity. The agent for treating inflammatory diseases according to claim 4, wherein the agent for treating inflammatory diseases is useful for any one inflammatory disease selected from the group consisting of chronic inflammation, rheumatoid arthritis, colitis, asthma and psoriasis.