KR100454649B1 - Seleno Compounds Containing Nitrone Moiety, Their Preparation and Their Therapeutic Uses - Google Patents

Abstract

The present invention relates to novel selenoid compounds containing nitrons, methods for their preparation, and various medical dysfunctions and diseases caused by reactive oxygen species (ROS), in particular stroke, Parkinson's disease. disease) and its use for the treatment and / or prevention of Alzheimer's disease. Compounds of the present invention have a lipid peroxidation (LPO) inhibitory activity that is comparable to or better than that of the reference compound. They are less toxic and more soluble, but they effectively inhibit cerebral neuronal cell death by free radicals (ROS) and exhibit neuroprotective effects against ischemic neurodegeneration.

Description

Seleno Compounds Containing Nitrone Moiety, Their Preparation and Their Therapeutic Uses

According to Harman's free-radical theory of aging, a continuous oxidative attack is a condition of "oxidative stress", or pro-oxidant. In excessive condition, the protection system is imbalanced. Such oxidative attacks result in innumerable molecular modifications, particularly in complex unsaturated membranes, proteins and nucleic acids. Human and animal subjects have a variety of defense mechanisms that work cooperatively with each other. These defense mechanisms are found in enzyme systems (superoxide dismutase, catalase and glutathione peroxidase) or non-enzymatic systems (such as vitamins E and C, which allow physiological control of free-radical activity). However, as aging progresses, the protective action is not very ineffective, but gradually diminished, especially with the decrease in activity of many enzymes, including enzymes involved in this protective mechanism. In conclusion, some diseases associated with aging, such as atherosclerosis, cataract, non-insulin-dependent diabetes, cancer, or chronic neurodegenerative disorders Many studies in the field have demonstrated that such diseases are associated with "oxidative stress" conditions.

The central nervous system is particularly sensitive to "oxidative stress" because of its high oxygen consumption, relatively low antioxidant defense systems and high levels of iron in some cerebral areas. This supports the fact that "oxidative stress" is one of the major causes of cerebral aging, as well as acute central nervous system disorders such as stroke and cerebral neurodegenerative diseases such as Parkinson's disease, Alzheimer's disease and basal ganglia degeneration. . The incidence of CNS neurological diseases is increasing worldwide, and stroke is the third leading cause of death after cardiovascular disease and malignancy (Parnetti, L. et al., Drug, 53: 752). 1997)).

Antioxidants that protect brain nerve cells from oxidative stress include vitamin E derivatives such as Trolox (J. Med. Chem., 38: 453 (1995)), glutathione peroxidase (hereinafter referred to as GPx). Mimetics (see Daiichi Pharmaceutical Co., Ltd., Annual Report (1999); WO 9808831; USP 5008394; J. Am. Chem. Soc., 119: 2079-2083 (1997); Adv. Pharmacol., 38: 229 (1996)), superoxide dismutase (SOD) mimetics (USP 5827880), spin trapping agents (J. Med. Chem., 39: 4988 ( 1996); USP 5475032).

In the case of GPx mimetics, it is a compound synthesized based on the selenocysteine site, which makes an important contribution at the GPx active site. Ebselen, the most representative GPx mimetic, has very low toxicity and shows good results in preclinical and clinical trials, and is especially of interest as a treatment for stroke. However, the water solubility is so low that its use as a drug is limited.

In the case of the spin trapping agent, development of an antioxidant can be achieved if it can sufficiently capture harmful free radicals in vivo. Such spin trapping agents include α-phenyl-N-t-butylnitron (PBN), and various derivatives of PBN are currently being developed. In general, nitron groups increase the solubility of the compound in water, but have little inhibitory effect on lipid peroxidation under laboratory conditions, and do not protect brain cells in vivo. (Fevig, Thomas L. et al., J. Med. Chem., 39: 4988-4996 (1996)).

Summary of the Invention

Accordingly, the present inventors have diligently researched to solve the above-mentioned problems, and as a result, the novel compound developed by introducing the nitron portion of the spin trapping agent into the epsxlen, a GPx mimetic, has excellent water solubility and low toxicity, as well as peroxidase. ) And at the same time have an anti-oxidant activity that effectively prevents and cures the death of brain cells with low toxicity, and confirms that it can be used as an effective drug in treating and preventing the death of brain cells. It was completed.

It is a first object of the present invention to provide a novel form of antioxidant that incorporates a spin trapping moiety in a GPx mimetic.

It is a second object of the present invention to provide a method for preparing an electric antioxidant.

A third object of the present invention includes an electric antioxidant as an active ingredient, pharmacologically useful for the treatment and / or prevention of various medical dysfunctions and diseases caused by active oxygen (ROS), in particular stroke, Parkinson's disease and Alzheimer's disease It is to provide a composition.

A fourth object of the present invention is to provide a method for treating an individual suffering from a disease in need of an antioxidant, in particular acute and progressive neurodegenerative diseases, comprising administering an electropharmacological preparation.

The present invention relates to a novel seleno compound containing nitrone, a preparation method thereof, and a pharmaceutical composition containing the new novel compound as an active ingredient. More specifically, the present invention relates to novel selenoid compounds containing nitrons, methods for their preparation, and various medical dysfunctions and diseases caused by reactive oxygen species (ROS), in particular strokes. , Parkinson's disease and Alzheimer's disease for their use for the treatment and / or prophylaxis.

The objects and advantages of the present invention and other objects and advantages will become more apparent from the following drawings and detailed description.

Figure 1 is a graph showing the result of treatment with a combination of Epsellene and Fe ²⁺ toxin.

Figure 2 is a graph showing the result of treating the compound synthesized in Example 1 and Fe ²⁺ toxin together.

Figure 3 is a graph showing the result of treating the compound synthesized in Example 2 and Fe ²⁺ toxin together.

Figure 4 is a graph showing the result of treating the compound synthesized in Example 5 and Fe ²⁺ toxin together.

5 is a graph showing the result of treating the compound synthesized in Example 7 with Fe ²⁺ toxin.

Figure 6 is a graph showing the extent of cell damage as the treatment concentration of eepselen increased.

7 is a graph showing the degree of cell damage as the treatment concentration of the compound synthesized in Example 1 increases.

8 is a graph showing the degree of cell damage as the treatment concentration of the compound synthesized in Example 2 increases.

9 is a graph showing the extent of cell damage with increasing treatment concentration of the compound synthesized in Example 5. FIG.

10 is a graph showing the extent of cell damage as the treatment concentration of the compound synthesized in Example 7 increases.

Fig. 11 (A) is a graph showing the degree of healing of cell damage in the case of treating the compound of the present invention after ischemia.

Fig. 11 (B) is a photomicrograph showing the degree of healing of cell damage when the compound of the present invention is treated after ischemia.

In a first embodiment, the present invention provides a novel antioxidant of formula (I) having the dual function of peroxidase activity and free radical capture activity:

(I)

Where

R ₁ and R ₂ are the same as or different from each other and are hydrogen, halogen, C _1-4 alkyl, C _1-4 alkoxy, hydroxy, trifluoromethyl, nitro or R ₁ and R ₂ together represent methylenedioxy. Forming;

L is phenyl, C _1-4 alkylphenyl, or furanyl, oxazolyl, isoxazolyl, thiophenyl, thiazolyl, isothiazolyl, pyrrolyl, imidazolyl, pyrazolyl, thiadiazolyl, pyridyl, Unsaturated with 1 to 4 nitrogen, oxygen or sulfur heteroatoms selected from the group consisting of pyrimidinyl, pyrazinyl, pyridazinyl, benzothiazolyl, benzoimidazolyl, benzotriazolyl, triazinyl and triazolyl Or a saturated heterocyclic group, wherein the heterocyclic group is halogen, C _1-2 alkyl, C _1-4 alkoxy, C _1-4 alkylthio, hydroxy, mercapto, trifluoromethyl, nitro, phenyl, nitrile , Carboxy or C _1-4 alkoxycarbonyl which may be substituted one or two times the same or different); And,

R ₃ is isopropyl or t-butyl.

In this context, preferred compounds include

R ₁ and R _{2, which} are the same or different, independently of one another, are hydrogen, fluorine, chlorine, bromine, hydroxy, methyl, ethyl, methoxy, trifluoromethyl, nitro or methylenedioxy;

R ₃ is isopropyl or t-butyl; And,

L is phenyl, methylphenyl, ethylphenyl, or furanyl, oxazolyl, thiophenyl, thiazolyl, pyrrolyl, imidazolyl, pyrazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, benzothiazolyl From triazinyl and triazolyl Unsaturated or saturated heterocyclic groups having 1 to 4 nitrogen, oxygen or sulfur heteroatoms selected, wherein the heterocyclic group is fluorine, chlorine, bromine, methyl, ethyl, butyl, methoxy, ethoxy, methyl Mercapto, ethyl mercapto, hydroxy, mercapto, trifluoromethyl, nitro, phenyl, nitrile, carboxy or methoxycarbonyl and ethoxycarbonyl one or two times the same or different May be substituted).

More preferred compound,

R ₁ and R _{2, which} are the same or different, independently of one another are hydrogen, fluorine, chlorine, methyl, methoxy, trifluoromethyl, nitro or methylenedioxy;

L is 1-4 selected from the group consisting of phenyl, methylphenyl, ethylphenyl, or furanyl, oxazolyl, thiophenyl, thiazolyl, pyrrolyl, imidazolyl, pyridyl, pyrimidinyl and benzothiazolyl Or saturated heterocyclic groups having two nitrogen, oxygen or sulfur heteroatoms wherein the heterocyclic group is fluorine, chlorine, bromine, methyl, methoxy, ethoxy, methylmercapto, hydroxy, mercapto, nitro, Phenyl, nitrile, carboxy or methoxycarbonyl and ethoxycarbonyl, which may be substituted one or two times the same or different); And,

R ₃ is a derivative that is isopropyl or t-butyl.

Compounds of the present invention have lipid peroxidation (LPO) inhibitory activity similar to or better than that of the comparable S-PBN and eepselen. They have lower toxicity and higher water solubility, but effectively inhibit cerebral neuronal cell death by free radicals and exhibit neuroprotective effects on ischemic neuronal degeneration.

Compounds of the present invention, in particular, compounds synthesized in Example 5 below, have very low toxicity at LD ₅₀ ≧ 7,000 mg / kg orally when administered in rats and ≧ 800 mg / kg when administered intraperitoneally. Thus, one of the advantages of the present invention is that the novel compounds can be administered at much higher levels than any other known antioxidants such as eepselen (LD ₅₀ values of eepselen obtained from mice are determined by oral administration). More than 6,810 mg / kg and more than 740 mg / kg when intraperitoneally administered Similarly, the LD ₅₀ value of espele obtained in rats was more than 6,810 mg / kg when administered orally and intraperitoneally 580 mg / kg). Thus, administration of large amounts of the novel compounds immediately after stroke or other trauma can significantly reduce oxidative damage.

In a second embodiment, the present invention provides a process for the preparation of a compound of formula (I) as shown in the following reaction pathway:

And is labeled as "1", has an appropriate linker, and then by reacting the aldehyde with an alkyl hydroxylamine (alkylhydroxylamines, R ₃ NHOH) the amino group is protected to give the nitrone labeled with "2", and by reacting this protecting group deionized Obtain a free amine nitron as indicated by " 3 ". Preferably, the alkylhydroxylamine is used without separating what is produced by reacting nitroalkane, zinc and acetic acid in one reactor. When the protecting group is removed, it is preferable to use trifluoroacetic acid when the protecting group is tert-butoxycarbonyl, or LiOH when it is acetyl.

The free amine of the compound designated as " 3 " is added to o-chloroselenobenzoyl chlorides, " 4 " in the presence of an excess of base or organic base, more preferably triethylamine. And a seleno compound containing the nitron portion of formula (I).

In a third embodiment, the present invention provides a pharmaceutical composition comprising a pharmaceutically acceptable carrier and one compound of formula (I) or a pharmaceutically acceptable salt thereof in a pharmaceutically effective amount.

In a fourth embodiment, the present invention provides a method of treating an individual comprising administering an electropharmaceutical composition to a subject suffering from a disease in need of antioxidants, particularly acute and progressive neurodegenerative diseases.

As mentioned above, the compounds of the present invention have proven to be effective antioxidants that mitigate various effects due to free radicals. These compounds are useful as therapeutic agents to treat and / or prevent a wide range of medical dysfunctions and diseases, including but not limited to acute central nervous system (CNS) disorders and neurodegenerative diseases.

As a pharmaceutical, the compounds of the present invention are generally administered in the form of a pharmacological composition comprising at least one active compound of the present invention and comprising a pharmaceutically acceptable carrier or carrier suitable for use in pharmacological preparations.

In general, the compounds of the present invention are administered in a pharmaceutically effective amount. The amount of the compound actually administered will usually be determined by the physician taking into account relevant conditions, including the subject disease, the route of administration chosen, the compound actually administered, age, weight and individual patient's response, the severity of symptoms, and the like. Dosages range from 10 mg to 500 mg once or several times daily.

The pharmaceutical compositions of the present invention can be administered by a variety of routes including oral, rectal, transdermal, subcutaneous, intravenous, intramuscular and nasal. The pharmaceutical composition of the present invention is preferably formulated in one of the injectable or oral compositions depending on the intended route of delivery.

Compositions for oral administration may take the form of bulk liquid dilutions or suspensions, or bulk powders. More generally, however, it is provided in unit dosage form to facilitate accurate dosing. The term "unit dosage forms" refers to completely separate units suitable for one dose to be administered to patients and other mammals, each of which is calculated and determined in advance to produce the desired therapeutic effect. Amounts of the active substance are in combined form with appropriate pharmaceutical excipients. Common unit dosage forms include ampules or syringes in which the liquid composition is prefilled and measured, or in the case of solid compositions, pills, tablets, capsules and the like. In such compositions the seleno compounds containing the nitron moiety of the invention are usually minor components (about 0.1 of the total weight, including other residues which treat the acid, which are various carriers or carriers and useful for making the desired dosage form). To 50% or preferably about 1 to 40%).

Liquid forms suitable for oral administration may include suitable aqueous or non-aqueous carriers with buffers, suspensions and dispersants, colorants, spices, and the like. Solid preparations may include, for example, the following components or compounds of similar nature: binders such as microcrystalline cellulose, gum tragacanth or gelatin; Excipients such as starch or lactose; Disintegrating agents such as alginic acid, Primogel or corn starch; Lubricants such as magnesium stearate; Glidants, such as colloidal silicon dioxide; Sweeteners such as sucrose or saccharin; Or a flavoring agent such as mint, methyl salicylate or orange flavoring.

Injectable compositions are generally based on sterile saline for injection or phosphate-buffered saline (PBS) or other injectable carriers known in the art. As before, the present compounds in such compositions are generally a minor component and usually comprise about 0.05-10% of the combined weight of the residue, such as an injectable carrier.

Electrical components for oral or injectable compositions are only representative. Other materials and methods of treatment and the like are disclosed in Remington's Pharmaceutical Sciences (Part 8 of Remington's Pharmaceutical Sciences, 17th edition, 1985, Mack Publishing Company, Easton, Pa.).

The compounds of the present invention can also be administered in a sustained release form or in a sustained release drug delivery system. Representative sustained release materials are disclosed in the Remingtons Pharmaceutical Sciences Additives column.

Hereinafter, the present invention will be described in more detail with reference to Examples. In accordance with the spirit of the invention the scope of the invention is not limited by these examples.

Example 1 2- [4- (N-isopropyl) nitronyl] phenyl-1,2-benzisoselenazole-3 (2H) -one

Synthesis of 8

First Step : Synthesis of 4-N- (1,1-dimethylethoxycarbonyl) aminobenzonitrile (2)

500 mg (4.23 mmol) of 4-aminobenzonitrile (1) and 1.90 g (8.70 mmol) of di-t-butyl dicarbonate (Boc ₂ O) were placed in a flask and heated at 110 ° C. for 6 hours. The reaction was cooled to room temperature and purified through short flash column chromatography (silica, CH ₂ Cl ₂ : Hex: EtOAc = 10: 10: 1) to give 630 mg (2.90 mmol) of a white solid compound (2 ) Was obtained in a yield of 68%.

¹ H NMR (CDCl ₃ ): δ 7.58 (d, J = 8.7 Hz, 2H), 7.48 (d, J = 8.7 Hz, 2H),

6.65 (br s, 1 H), 1.53 (s, 9 H).

Second Step : Synthesis of 4-N- (1,1-dimethylethoxycarbonyl) aminobenzaldehyde (3)

After dissolving 600 mg (2.75 mmol) of nitrile (2) in CH ₂ Cl ₂ (8 mL), 8.3 mL (8.3 mmol) of diisobutylaluminum hydride (DIBAL-H, 1.0M solution in toluene) at −78 ° C. Was added over 2 minutes. After about 1 hour 2 mL of MeOH was added slowly. After raising the reaction to room temperature, diethyl ether and 0.5N HCl solution were added, and the organic layer was separated. The aqueous solution was once again extracted with diethyl ether and the combined organic layers were washed with saturated NaHCO ₃ solution, dried over anhydrous Na ₂ SO ₄ , the organic solvent was removed under reduced pressure and the resulting residue was subjected to flash column chromatography (silica, Purification via CH ₂ Cl ₂ : Hex: EtOAc = 10: 10: 1 to 10: 10: 2) afforded 585 mg (2.64 mmol) of a white solid compound (3) in a yield of 96%.

¹ H NMR (CDCl ₃ ): δ 9.89 (s, 1H), 7.83 (d, J = 8.6 Hz, 2H), 7.53 (d, J =

8.6 Hz, 2H), 6.70 (br s, 1 H), 1.55 (s, 9 H).

Third Step : N-isopropyl-α- [4-N- (1,1-dimethylethoxycarbonylamino) phenyl] nit

Synthesis of Ron 5

422 mg (1.90 mmol) of compound 3, 680 mg (7.63 mmol) of 2-nitropropane (4) and 745 mg (11.40 mmol) of zinc were added to a flask containing 95% ethanol (8 mL) and 0.87 mL (0. 15.20 mmol) of acetic acid was added slowly. The reaction was raised to room temperature, stirred for 6 hours, CH ₂ Cl ₂ was added, filtered through a Celite pad and concentrated under reduced pressure. The resulting residue was purified via flash column chromatography (silica, CH ₂ Cl ₂ : EtOAc = 1: 2) to yield 500 mg (1.80 mmol) of white solid compound (5) in 95% yield.

¹ H NMR (CDCl ₃ ): δ 8.21 (d, J = 8.9 Hz, 2H), 7.42 (d, J = 8.9 Hz, 2H),

7.37 (s, 1 H), 6.61 (br s, 1 H), 4.19 (septet, J = 6.5 Hz,

1H), 1.52 (s, 9H), 1.50 (d, J = 6.5 Hz, 6H).

Fourth Step : Synthesis of N-isopropyl-α- (4-aminophenyl) nitron (6)

320 mg (1.15 mmol) of compound 5 were dissolved in CH ₂ Cl ₂ (10 mL), and 1 mL of trifluoroacetic acid was slowly added at 0 ° C. The reaction was slowly raised to room temperature and stirred for 16 hours. The solution was concentrated and then diluted with CH ₂ Cl ₂ and saturated NaHCO ₃ . NaCl was added thereto to make the solution saturated, and the organic layer was separated. After the aqueous layer was extracted three times again with CH ₂ Cl ₂ , the combined organic layers were dried over anhydrous Na ₂ SO _4, and concentrated under reduced pressure. The resulting residue was purified by flash column chromatography (silica, CH ₂ Cl ₂ : EtOAc: MeOH = 5: 5: 1) to give 132 mg (0.74 mmol) of yellow solid Compound 6 in 64% yield. .

¹ H NMR (CDCl ₃ ): δ 8.12 (d, J = 7.0 Hz, 2H), 7.27 (s, 1H), 6.68 (d, J =

7.0 Hz, 2H), 4.19 (septet, J = 6.5 Hz, 1H), 1.51 (d, J =

6.5 Hz, 6H);

¹³ C NMR (CDCl ₃ ): δ 149.08, 132.69, 131.04, 121.39, 114.66, 67.13,

21.23.

Fifth Step : 2- [4- (N-isopropyl) nitronyl] phenyl-1,2-benzisoselenazole-3 (2H) -one

Synthesis of 8

2 of 75mg (0.42mmol) 200mg (0.79mmol) was dissolved in CH ₂ Cl ₂ (1.5mL) to a solution of compound 6 with _{1.0mL (7.17mmol) CH 2 Cl 2} (3mL) containing triethylamine of -Chlorocarbonyl-benzeneselenyl chloride (7) was added slowly at 0 ° C. After stirring for 4 hours at room temperature, the reaction was concentrated under reduced pressure, and the resulting residue was purified by flash column chromatography (silica, CH ₂ Cl ₂ : EtOAc = 3: 1, containing 0 to 10% methanol). 83 mg (0.23 mmol) of pale yellow solid Compound 8 were obtained in a yield of 55%.

¹ H NMR (CDCl ₃ : CD ₃ OD = 4: 1): δ 8.33 (d, J = 8.8 Hz, 2H), 8.08 (dd, J =

7.8 and 0.7 Hz, 1H), 7.83 (d, J = 8.0 Hz, 1H), 7.77

(dd, J = 8.8 and 1.9 Hz, 2H) 7.75 (t, J = 7.8 Hz, 1H),

7.60 (s, 1 H), 7.48 (t, J = 7.8 Hz, 1 H), 7.39 (s, 1 H),

4.28 (septet, J = 6.5 Hz, 1H), 1.52 (d, J = 6.5 Hz,

6H);

¹³ C NMR (CDCl ₃ : CD ₃ OD = 4: 1): δ 166.30, 141.11, 138.27, 133.14, 132.67,

129.92, 128.78, 127.86, 127.67, 126.40, 124.37, 124.21,

67.66, 20.44.

Example 2 : 2- [3- (N-isopropyl) nitronyl] phenyl-1,2-benzisoselenazole-3 (2H) -one

Synthesis of 15

First Step : Sum of Ethyl 3-N- (1,1-dimethylethoxycarbonyl) aminobenzoate (10)

castle

150 mL of 1,4-dioxane / H ₂ O (1: 1 v / v) containing 5.0 g (30.27 mmol) of ethyl 3-aminobenzoate (9) and 17 mL (0.12 mol) of triethylamine 16.52 g (75.67 mmol) of di-t-butyl dicarbonate (Boc ₂ O) was added thereto, followed by stirring at room temperature for 13 hours. Water and diethyl ether were added to the reaction mixture, the organic layer was separated, washed with saturated NaCl solution, and dried over anhydrous Na ₂ SO ₄ . Thereafter, the resulting residue was filtered and concentrated under reduced pressure, and the resulting residue was purified by washing with n-hexane to give 7.83 g (29.5 mmol) of the white solid compound 10 in a yield of 98%.

¹ H NMR (CDCl ₃ ): δ 7.90 (t, J = 1.7 Hz, 1H), 7.71 (m, 2H), 7.36 (t, J =

7.9 Hz, 1H), 6.63 (br s, 1H), 4.37 (m, 2H), 1.52 (s,

9H), 1.38 (t, J = 7.1 Hz, 3H).

Second Step : Synthesis of 3-N- (1,1-dimethylethoxycarbonyl) aminobenzyl alcohol (11)

To a solution of CH ₂ Cl ₂ (200 mL) containing 9.64 g (36.34 mmol) of ethyl benzoate 10, 109 mL of diisobutylaluminum hydride (DIBAL-H, 1.0 M solution in toluene) was added at -78 ° C. Minutes were added. After stirring for 3 hours at room temperature, 30 mL of MeOH was added slowly. After raising the reaction to room temperature, diethyl ether and 0.5N HCl solution were added, and the organic layer was separated. The solution was once again extracted with diethyl ether. The combined organic layer was washed with saturated NaHCO ₃ solution, dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica, CH ₂ Cl ₂ : Hex: EtOAc = 10: 10: 1 to 10: 10: 2) to afford 7.2 g (32.3 mmol) of compound 11 in 89% yield. It was.

¹ H NMR (CDCl ₃ ): δ 7.42 (s, 1H), 7.24 (m, 2H), 7.0 (t, J = 6.9 Hz, 1H),

6.56 (s, 1H), 4.64 (s, 2H), 1.53 (s, 9H)

Third Step : Synthesis of 3-N- (1,1-dimethylethoxy-carbonyl) aminobenzaldehyde (12)

A DMSO solution of 5.63mL 6.92mL (96.74mmol) was dissolved in octanoic CH ₂ Cl ₂ (60mL) to a solution of CH ₂ Cl ₂ (60mL) containing a chloride of a save (64.50mmol) was slowly added at -78 ℃. After 10 minutes, CH ₂ Cl ₂ (60 mL) containing 7.2 g (32.3 mmol) of compound 11 was slowly added and stirred for 30 minutes, followed by the slow addition of 34 mL of TEA. After raising the reaction to room temperature, CH ₂ Cl ₂ and water were added, the organic layer was separated, washed with saturated NaCl solution, dried over anhydrous Na ₂ SO ₄ , filtered and concentrated. The resulting solid was purified with n-hexane to give 6.55 g (29.60 mmol) of compound 12 in 92% yield.

¹ H NMR (CDCl ₃ ): δ 9.99 (t, J = 3.4 Hz, 1H), 7.92 (t, 1H), 7.64 (d, 1H),

7.62 (d, 1 H), 7.45 (t, 1 H), 6.70 (s, 1 H), 1.55 (s, 9 H).

Fourth Step : N-isopropyl-α- [3-N- (1,1-dimethylethoxycarbonyl) amino] phenyl ni

Synthesis of Tron 13

6.55 g (29.6 mmol) of Compound 12, 6.65 ml (74.03 mmol) of 2-nitropropane (4) and 6.78 g (103.65 mmol) of zinc were placed in a round bottom flask containing 95% ethanol (100 mL) at 0 ° C. Cooled. 11.9 mL (207.9 mmol) of acetic acid was slowly added while stirring, the temperature was raised to room temperature, and then stirred for 12 hours. CH ₂ Cl ₂ was added to the reaction, filtered through a pad of celite and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica, CH ₂ Cl ₂ : EtOAc = 1: 2) to 7.0 g (21.3 mmol). Compound 13 (mp: 189-191C) was obtained in a yield of 72%.

¹ H NMR (CDCl ₃ ): δ 8.37 (s, 1H), 7.8 (d, J = 7.7 Hz, 1H), 7.5 (d, J =

7.7 Hz, 1H), 7.42 (s, 1H), 7.33 (t, J = 7.8 Hz, 1H),

6.60 (s, 1 H), 4.19 (septet, J = 6.5 Hz, 1 H), 1.53 (s,

9H), 1.48 (d, J = 6.5 Hz, 6H).

Fifth Step : Synthesis of N-isopropyl-α-3-aminophenyl nitron (14)

20 mL of trifluoroacetic acid was slowly added to a solution of CH ₂ Cl ₂ (200 mL) containing 5.0 g (17.96 mmol) of compound 13. The reaction was raised to room temperature and stirred for 16 hours. The solution was concentrated and then diluted with CH ₂ Cl ₂ and saturated NaHCO ₃ solution. NaCl was added thereto to make the solution saturated, and the organic layer was separated. After extracting the aqueous layer three times with CH ₂ Cl ₂ , the combined organic layers were dried over anhydrous Na ₂ SO ₄ and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica, CH ₂ Cl ₂ : EtOAc: MeOH = 5: 5: 1) to give 2.1 g (11.78 mmol) of yellow solid Compound 14 (mp: 103-106 ° C.) 66%. Obtained in the yield.

¹ H NMR (CDCl ₃ ): δ 8.12 (t, 1H), 7.30 (s, 1H), 7.16 (d, 2H), 6.74 (m,

1H), 4.18 (septet, J = 6.5 Hz, 1H), 3.74 (br s, 2H),

1.49 (d, J = 6.5 Hz, 6H);

¹³ C NMR (CDCl ₃ ): δ 147.11, 132.80, 131.88, 129.52, 119.87, 117.38,

114.77, 68.10, 21.24.

Sixth Step : 2- [3- (N-isopropyl) nitronyl] phenyl-1,2-benzisoselenazole-3 (2H) -one

Synthesis of 15

In a solution of CH ₂ Cl ₂ (3 mL) containing 50 mg (0.28 mmol) of Compound 14 and 0.8 mL (5.62 mmol) of triethylamine, 178 mg (0.70 mmol) of 2-chlorocarbonyl-benzeneselenyl chloride ( CH ₂ Cl ₂ (1.5 mL) containing 7) was slowly added at 0 ° C. After stirring for 4 hours at room temperature, the reaction was concentrated under reduced pressure. The residue was purified by flash column chromatography (silica, CH ₂ Cl ₂ : EtOAc = 3: 1, containing 0 to 10% methanol) to 60 mg (0.17 mmol) of pale yellow solid compound 15 (mp: 94-98 ° C.). ) Was obtained in a yield of 60%.

¹ H NMR (CDCl ₃ ): δ 8.65 (m, 1H), 8.09 (m, 2H), 7.81 (m, 1H), 7.66 (m,

2H) 7.37 (m, 3H), 4.23 (septet, J = 6.5 Hz, 1H), 1.52

(d, J = 6.5 Hz, 6H);

¹³ C NMR (CDCl ₃ ): δ 166.23, 139.77, 138.18, 133.00, 132.21, 131.79,

129.69, 127.95, 127.12, 126.96, 126.90, 125.17, 124.30,

68.53, 67.48, 21.33.

Example 3 5-Chloro-2- [3- (N-isopropyl) -nitronyl] phenyl-1,2-benzisoselenazole

Synthesis of -3 (2H) -on (17)

290 mg (1.01 mmol) of 4-chloro-2-chlorocarbonylbenzeneselenyl chloride (16) and 100 mg (0.56 mol) of N-isopropyl in a similar manner to the method disclosed to obtain compound 8 in Example 1 40 mg (0.10 mmol) of yellow solid compound 17 were obtained from -α-3-aminophenylnitron (14) in a yield of 18%.

¹ H NMR (CDCl ₃ : CD ₃ OD = 4: 1): δ 8.63 (t, J = 1.7 Hz, 1H), 8.02 (d, J =

2.2 Hz, 1H), 8.00 (d, J = 7.9 Hz, 1H), 7.81 (dd, J =

7.9 and 2.3 Hz, 1H), 7.81 (d, J = 8.5 Hz, 1H), 7.61 (s,

1H), 7.61 (dd, J = 8.5 and 2.3 Hz, 1H), 7.53 (t, J = 7.9

Hz, 1H), 4.28 (septet, J = 6.5 Hz, 1H), 1.50 (d, J =

6.5 Hz, 6H)

¹³ C NMR (CDCl ₃ : CD ₃ OD = 4: 1): δ 166.30, 141.11, 138.27, 133.14, 132.67,

129.92, 128.77, 127.86, 127.67, 126.45, 124.37, 124.21,

67.66, 20.44.

Example 4 5-Methyl-2- [3- (N-isopropyl) nitronyl] phenyl-1,2-benzisoselenazole-

Synthesis of 3 (2H) -one (19)

380 mg (1.40 mmol) of 4-methyl-2-chlorocarbonylbenzeneselenyl chloride (18) and 100 mg (0.56 mmol) of N-isopropyl in a similar manner to the method disclosed for obtaining compound 8 in Example 1 40 mg (0.20 mmol) of a yellow solid compound 19 (mp: 197 to 201C) was obtained from -α-3-aminophenylnitron (14) in a yield of 30%.

¹ H NMR (CDCl ₃ ): δ 8.60 (s, 1H), 8.09 (d, J = 7.7 Hz, 1H), 7.90 (s, 1H),

7.81 (d, J = 8.0 Hz, 1H) 7.56 (d, J = 8.0 Hz, 1H), 7.44

(m, 3H), 4.23 (septet, J = 6.5 Hz, 1H), 2.47 (s, 3H),

1.51 (d, J = 6.5 Hz, 6H);

¹³ C NMR (CDCl ₃ ): δ 166.25,139.91, 137.07, 134.76, 134.39, 132.19,

131.78, 129.73, 129.71, 127.94, 127.09, 126.85, 125.14,

123.96, 68.51, 60.79, 21.41, 14.59.

Example 5 2- [4- (N-isopropyl) nitronyl] thiazol-2-yl-1,2-benzisoselenazole-

Synthesis of 3 (2H) -one (26)

First Step : Ethyl 2-N- (1,1-dimethylethoxy-carbonyl) aminothiazole-4-carboxylate

Synthesis of meth 21

6.05 g (35.13 mmol) of aminothiazole 20 and 26.84 g (0.12 mmol) of di-t-butyl dicarbonate (Boc ₂ O) were placed in a flask and heated to 110 ° C. for 24 hours. The reaction was cooled to room temperature and purified by flash column chromatography (silica, CH ₂ Cl ₂ : Hex: EtOAc = 10: 6: 3) to yield 7.13 g (26.18 mmol) of white solid compound 21 in 74.5% yield. It was.

¹ H NMR (CDCl ₃ ): δ 8.21 (br s, 1H), 7.78 (s, 1H), 4.38 (q, J = 7.1 Hz,

2H), 1.54 (s, J = 7.1 Hz, 9H), 1.38 (t, 3H).

Second step : 2-N- (1,1-dimethylethoxy-carbonyl) aminothia (from ester 21)

Synthesis of Sol-4-Carbaldehyde (22)

To a solution of CH ₂ Cl ₂ (75 mL) containing 7.0 g (25.71 mmol) of ethyl ester 21 was added 77 mL of diisobutylaluminum hydride (DIBAL-H, 1.0 M solution in toluene) for 20 minutes at -78 ° C. Was added. After stirring for 3 hours at that temperature, 30 mL of MeOH was added slowly and the temperature was raised to room temperature. After diethyl ether and 0.5N HCl solution were added to separate the organic layer, the aqueous layer was once again extracted with diethyl ether. The combined organic layers were washed with saturated NaHCO ₃ solution, dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was subjected to flash column chromatography (silica, CH ₂ Cl ₂ : Hex: EtOAc = 10: 6: Purification with 3 to CH ₂ Cl ₂ : EtOAc = 1: 1) afforded 1.90 g (8.32 mmol, yield 32.4%) of solid aldehyde 22 and 3.5 g (15.20 mmol) of liquid alcohol 23 in a yield of 59.0%.

Aldehyde 22:

¹ H NMR (CDCl ₃ ): δ 9.88 (s, 1H), 8.83 (br s, 1H), 8.82 (s, 1H), 1.58 (s,

9H).

Alcohol 23:

¹ H NMR (CDCl ₃ ): δ 6.75 (s, 1H), 4.58 (s, 2H), 1.58 (s, 9H)

Step 2-1 : 2-N- (1,1-dimethylethoxy-carbonyl) aminothi (from Alcohol 23)

Synthesis of azole-4-carbaldehyde (22)

302 mg (0.86 mmol) of tetrapropylammonium perruthenate (TPAP), 3.11 g (26.547 mmol) of NMO (N-methylmorpholine N-oxide) in CH ₂ Cl ₂ (50 mL) containing 2.04 g (8.597 mmol) of alcohol 23 ) And 16 g (2 g / 1 mmol of 4 g) molecular sieve were added and stirred at room temperature for 2 hours, after which the reaction was filtered through a pad of celite and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica, CH ₂ Cl ₂ : Hex: EtOAc = 10: 6: 3) to give 950 mg (4.0 mmol) of aldehyde 22 in a yield of 46.5%.

Third Step : N-isopropyl-α- [2-N- (1,1-dimethylethoxycarbonyl) aminothiazole-4-

Synthesis of Nitron (24)

2.22 g (9.72 mmol) of compound 22, 3.47 g (33.65 mmol) 2-nitropropane (4) and 2.54 g (38.84 mmol) of zinc were placed in a round bottom flask containing 95% ethanol (50 mL) at 0 ° C. Cooled. 4.67 g (77.77 mmol) of acetic acid was slowly added thereto while stirring. The reaction was raised to room temperature and stirred for 6 hours. CH ₂ Cl ₂ was added to the reaction, filtered through a pad of celite and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica, Hex: EtOAc = 1: 1) to afford 2.51 g (8.80 mmol) of compound 24 in 90.5% yield.

¹ H NMR (CDCl ₃ ): δ 8.71 (s, 1H), 7.63 (s, 1H), 4.21 (septet, J = 6.6 Hz,

1H), 1.55 (s, 9H), 1.49 (d, J = 6.6 Hz, 6H).

Fourth Step : Synthesis of N-isopropyl-α- (2-aminothiazol-4-yl) nitron (25)

To a solution of CH ₂ Cl ₂ (30 mL) containing 2.44 g (8.55 mmol) of compound 24, 3.3 mL of trifluoroacetic acid was slowly added at 0 ° C. The reaction was raised to room temperature and stirred for 16 hours. The solution was concentrated and then diluted with CH ₂ Cl ₂ and saturated NaHCO ₃ solution. NaCl was added thereto to make the solution saturated, and the organic layer was separated. The aqueous layer was extracted three times with CH ₂ Cl ₂ , and the combined organic layers were dried over anhydrous Na ₂ SO ₄ and concentrated under reduced pressure. The residue was subjected to flash column chromatography (silica, CH ₂ Cl ₂ : EtOAc: MeOH = 5). : 5: 1) to give 1.6 g (8.46 mmol) of a yellow solid compound 25 in 99% yield.

¹ H NMR (CDCl ₃ ): δ 8.38 (s, 1H), 7.59 (s, 1H), 5.61 (b, 2H), 4.16

(septet, J = 6.5 Hz, 1H), 1.46 (d, J = 6.5 Hz, 6H).

Fifth Step : 2- [4- (N-isopropyl) nitronyl] thiazol-2-yl-1,2-benzisoselenazole-

Synthesis of 3 (2H) -on (26)

In a solution of CH ₂ Cl ₂ (15 mL) containing 100 mg (0.53 mmol) of Compound 25 and 0.74 mL (5.29 mmol) of triethylamine, 220 mg (0.866 mmol) of 2-chlorocarbonyl-benzeneselenylchloride ( CH ₂ Cl ₂ (5 mL) containing 7) was added slowly at 0 ° C. After stirring for 1 hour at room temperature, the reaction was concentrated under reduced pressure. The residue was recrystallized (MeOH / CH ₂ Cl ₂ ) to give 70 mg (0.19 mmol) of pale yellow solid Compound 26 in 37% yield.

¹ H NMR (CD ₃ OD): δ 8.82 (s, 1H), 8.15 (s, 1H), 8.05 (d, J = 7.8 Hz, 1H),

8.03 (d, J = 8.0 Hz, 1H), 7.75 (t, J = 7.3 Hz, 1H), 7.53

(t, J = 7.4 Hz, 1H), 4.43 (septet, J = 6.8 Hz, 1H), 1.50

(d, J = 6.8 Hz, 6H).

Example 6 : 2- [4- (Nt-butyl) nitronyl] thiazol-2-yl-1,2-benzisoselenazole-3 (2H)-

Synthesis of On

First step : Nt-butyl-α- [2-N- (1,1-dimethylethoxycarbonyl) aminothiazol-4-yl]

Synthesis of Nitron (28)

2.0 g (8.76 mmol) of 22, 5.42 g (52.57 mmol) of 2-methyl-2-nitropropane (27) and 2.86 g (43.81 mmol) of zinc in a round bottom flask containing 95% ethanol (50 mL) Put and cooled to 0 ° C. 4.21 g (70.11 mmol) of acetic acid was slowly added while stirring and the temperature was raised to room temperature, followed by stirring for 6 hours. CH ₂ Cl ₂ was added to the reaction, filtered through a pad of celite and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica, Hex: EtOAc = 1: 1) to give 1.28 g (4.28 mmol) of a yellow solid. Compound 28 was obtained with a yield of 49%.

¹ H NMR (CDCl ₃ ): δ 9.9 (br s, 1 H), 8.82 (s, 1 H), 7.87 (s, 1 H), 1.60 (s,

9H), 1.54 (s, 6H);

¹³ C NMR (CDCl ₃ ): δ 159.54, 152.35, 141.53, 125.78, 117.31, 82.83,

70.33, 28.27, 28.21.

Second Step : Synthesis of Nt-Butyl-α- (2-aminothiazol-4-yl) nitron (29)

To a solution of CH ₂ Cl ₂ (10 mL) containing 200 mg (0.668 mmol) of compound 28, 381 mg of trifluoroacetic acid was slowly added at 0 ° C., the reaction was allowed to rise to room temperature, and stirred for 14 hours. The solution was concentrated and then diluted with CH ₂ Cl ₂ and saturated NaHCO ₃ solution. NaCl was added thereto to make the solution saturated, and the organic layer was separated. The aqueous layer was extracted three times with CH ₂ Cl ₂ , and the combined organic layers were dried over anhydrous Na ₂ SO ₄ and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica, EtOAc) to give 111 mg (0.56 mmol) of the residue. A yellow solid compound 29 was obtained in 83% yield.

¹ H NMR (MeOD): δ 8.29 (s, 1 H), 7.82 (s, 1 H), 4.91 (s, 2H), 1.54 (s,

9H);

¹³ C NMR (MeOD): δ 168.76, 141.94, 127.57, 114.24, 70.23, 27.20.

Third Step : 2- [4- (Nt-Butyl) nitronyl] thiazol-2-yl-1,2-benzisoselenazole-3 (2H)

Synthesis of On-30

In a solution of CH ₂ Cl ₂ (15 mL) containing 100 mg (0.50 mmol) of compound 29 and 0.70 mL (5.02 mmol) of triethylamine, 180 mg (0.703 mmol) of 2-chlorocarbonyl-benzeneselenyl chloride ( CH ₂ Cl ₂ (5 mL) containing 7) was added at 0 ° C. After stirring for 1 hour at room temperature, the reaction was concentrated under reduced pressure. The residue was purified by flash column chromatography (silica, EtOAc: Hex = 1: 1) to give 67 mg (0.176 mmol) of pale yellow solid Compound 30 in 35% yield.

¹ H NMR (CDCl ₃ : CD ₃ OD = 10: 1): δ 8.80 (s, 1H), 8.04 (d, J = 7.6 Hz, 1H),

7.91 (s, 1 H), 7.60 (d, J = 7.86 Hz, 1 H), 7.61 (t, J =

7.2 Hz, 1H), 7.40 (t, J = 7.41 Hz, 1H), 1.56 (s, 9H);

¹³ C NMR (CDCl ₃ : CD ₃ OD = 10: 1): δ 165.38, 157.10, 140.74, 139.19, 133.71,

128.76, 127.05, 126.78, 124.72, 119.43, 70.54, 28.05.

Example 7 : 2- [4- (N-isopropyl) nitronyl] benzyl-1,2-benzisoselenazole-3 (2H) -one

Synthesis of 37

First step : methyl 4-N- (1,1-dimethylethoxycarbonyl) aminomethylbenzoate (32)

synthesis

In a solution of CH ₂ Cl ₂ (10 mL) containing 500 mg (2.48 mmol) of methyl 4-aminomethylbenzoate HCl salt (31), 753 mg (7.45 mmol) of TEA and 568 mg (2.60 mmol) of Boc ₂ O were added. Included CH ₂ Cl ₂ (1 mL) was added at 0 ° C. After 30 minutes, the reaction was raised to room temperature and stirred for 4 hours. CH ₂ Cl ₂ was then added to the reaction solution, the organic layer was washed with 0.1N HCl solution, dried over MgSO ₄ , filtered and concentrated under reduced pressure. The residue was subjected to flash column chromatography (silica, Hex: EtOAc = 1: 1). Purification with 1) gave 620 mg of compound 32 in 94% yield.

¹ H NMR (CDCl ₃ ): δ 7.58 (d, J = 8.2 Hz, 2H), 7.34 (d, J = 8.2 Hz, 2H),

4.90 (br s, 1 H), 4.37 (d, 2 H), 3.91 (s, 3 H), 1.46 (s,

9H);

¹³ C NMR (CDCl ₃ ): δ 167.02, 156015, 144042, 130.03, 129.23, 127.27,

79.91, 52.23, 44.43, 28.49

Second Step : Sum of 4-N- (1,1-dimethylethoxy-carbonyl) aminomethylbenzyl alcohol (33)

castle

To a solution of CH ₂ Cl ₂ (15 mL) containing 620 mg (2.34 mmol) of ethyl benzoate 32, 7.01 mL of diisobutylaluminum hydride (DIBAL-H, 1.0M solution in toluene) was added at -78 ° C. Added for minutes. After stirring for 3 hours at that temperature, 3 mL of MeOH was added slowly and the temperature was raised to room temperature. After diethyl ether and 0.5N HCl solution were added to separate the organic layer, the aqueous layer was extracted once again with diethyl ether. The combined organic layer was washed with saturated NaHCO ₃ solution, dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica, Hex: EtOAc = 2: 1) to give 520 mg (2.19 mmol) of compound 33 in 94% yield.

¹ H NMR (CDCl ₃ ): δ 7.32 (m, 4H), 4.80 (br s, 1 H), 4.68 (s, 2H), 4.31 (m,

2H), 1.46 (s, 9H);

¹³ C NMR (CDCl ₃ ): δ 156.6, 140.19, 138.30, 127.69, 127.33, 79.67, 64.95,

44.44, 28.49

Third step : 4-N- (1,1-dimethylethoxy-carbonyl) aminomethylbenzaldehyde (34)

synthesis

Is the 0.48mL (5.48mmol) octanoic CH ₂ Cl ₂ (2mL) containing the DMSO of 0.63mL (8.76mmol) to a solution of containing chloride save CH ₂ Cl ₂ (2mL) slowly at -78 ℃ It was. After 15 minutes, CH ₂ Cl ₂ (3 mL) containing 520 mg (2.19 mmol) of compound 33 was slowly added dropwise and the reaction stirred for 30 minutes. 2.5 mL of TEA was slowly added dropwise to the reaction and the temperature was raised to room temperature. Then, CH ₂ Cl ₂ and H ₂ O were added to separate the organic layer, and the organic layer was washed with saturated NaCl solution, dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica, Hex: EtOAc = 2: 1) to yield 510 mg (2.17 mmol) of compound 34 in 99% yield.

¹ H NMR (CDCl ₃ ): δ 9.99 (s, 1H), 7.85 (d, J = 7.9 Hz, 2H), 7.44 (d, J =

7.9 Hz, 2H), 4.95 (br s, 1H), 4.40 (d, 2H), 1.47 (s,

9H);

¹³ C NMR (CDCl ₃ ): δ 191.95, 156.01, 146.37, 135.24, 129.93, 127.53,

79.57, 44.13, 28.28

Fourth Step : N-isopropyl-α- [4-N- (1,1-dimethylethoxycarbonylamino) methylphenyl]

Synthesis of Nitron 35

500 mg (2.13 mmol) of compound 34, 0.44 mL (4.84 mmol) of 2-nitropropane (4) and 565 mg (8.64 mmol) of zinc were placed in a round bottom flask containing 95% ethanol (10 mL) and cooled to 0 ° C. . 0.83 mL of acetic acid was slowly added dropwise with stirring, and then the temperature was raised to room temperature and stirred for 6 hours. Then, CH ₂ Cl ₂ was added to the reaction, filtered through a pad of celite, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica, Hex: EtOAc = 1: 1) to give 540 mg (1.85 mmol) of compound 35 in 87% yield.

¹ H NMR (CDCl ₃ ): δ 8.21 (d, J = 8.2 Hz, 2H), 7.42 (s, 1H), 7.32 (d, J =

8.2 Hz, 2H), 4.86 (br s, 1H), 4.33 (m, 2H), 4.23

(septet, J = 6.5 Hz, 1H), 1.50 (d, J = 6.5 Hz, 6H), 1.45

(s, 9H);

¹³ C NMR (CDCl ₃ ): δ 156.00, 141.33, 131.80, 129.62, 128.78, 127.28,

79.42, 67.64, 44.36, 28.37, 20.83

Fifth Step : Synthesis of N-isopropyl-α- (4-aminomethyl-phenyl) nitron (36)

0.34 mL of trifluoroacetic acid was slowly added dropwise at 0 ° C. to a solution of CH ₂ Cl ₂ (3 mL) containing 200 mg (0.68 mmol) of compound 35, and the reaction mixture was then heated to room temperature and stirred for 6 hours. The solution was concentrated and then diluted with CH ₂ Cl ₂ and saturated NaHCO ₃ solution. NaCl was added thereto to make the solution saturated, and the organic layer was separated. After extracting the aqueous layer three times with CH ₂ Cl ₂ , the combined organic layers were dried over anhydrous Na ₂ SO ₄ and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica, EtOAc: MeOH = 9: 1 to 4: 1) to give 130 mg (0.68 mmol) of a yellow solid compound 36 in 99% yield.

¹ H NMR (CDCl ₃ ): δ 8.10 (d, J = 8.4 Hz, 2H), 7.45 (s, 1H), 7.34 (d, J =

8.4 Hz, 2H), 4.13 (septet, J = 6.54 Hz, 1H), 3.88 (s,

2H), 1.41 (d, J = 6.54 Hz, 6H);

¹³ C NMR (CDCl ₃ ): δ 137.05, 135.89, 132.38, 130.98, 130.05, 68.80,

43.92, 20.90

Sixth Step : 2- [4- (N-isopropyl) -nitronyl] benzyl-1,2-benzisoselenazole-3 (2H)-

Synthesis of On 37

138 mg (0.54 mmol) of 2-chlorocarbonylbenzene selene in a solution of CH ₃ CN (15 mL) and EtOH (1 mL) containing 80 mg (0.42 mmol) of Compound 36 and 0.29 mL (2.08 mmol) of triethylamine CH ₃ CN (4 mL) containing niyl chloride (7) was added slowly at 0 ° C. After stirring for 4 hours at room temperature, the reaction was concentrated under reduced pressure. The residue was purified by flash column chromatography (silica, EtOAc) to afford 70 mg (0.19 mmol) of pale yellow solid compound 37 in 45% yield.

¹ H NMR (CDCl ₃ ): δ 8.24 (d, J = 8.1 Hz, 2H), 8.04 (d, J = 7.9 Hz, 1H),

7.91 (s, 1 H), 7.87 (d, J = 6.4 Hz, 1 H), 7.63 (d, J = 6.8

Hz, 1H), 7.45 (t, J = 6.9 Hz, 1H), 7.37 (d, J = 8.1 Hz,

2H), 4.95 (s, 2H), 4.34 (septet, J = 6.3 Hz, 1H), 1.36

(d, J = 6.3 Hz, 6H);

¹³ C NMR (CDCl ₃ ): δ 140.12, 139.24, 131.99, 130.73, 129.20, 128.66,

128.06, 126.29, 125.79, 68.09, 48.21, 21.021

Example 8 7-nitro-2- [4- (N-isopropyl) nitronyl] phenyl-1,2-benzisoselenazole-

Synthesis of 3 (2H) -on (40)

First step : synthesis of 2-methylseleno-3-nitrobenzoic acid (38)

2.80 mL (4.47 mmol) of n-BuLi (1.6 M soln. In Hex.) Was added to a solution of anhydrous THF (15 mL) containing 500 mg (2.0 mmol) of 2-bromo-3-nitrobenzoic acid. It was slowly added dropwise at 占 폚. After 10 minutes, THF (5 mL) containing 383 mg (2.03 mmol) of dimethyl diselenide was added. After 30 minutes, the reaction was raised to room temperature and further stirred for 2 hours, after which ethyl acetate was added. The organic layer was washed with 1N HCl solution, dried over MgSO ₄ and concentrated under reduced pressure. 470 mg of crude residue was used for the next reaction without further purification.

¹ H NMR (CD ₃ OD): δ 7.91 (d, J = 7.85 Hz, 1H), 7.88 (d, J = 7.86 Hz, 1H),

7.56 (t, J = 7.85 Hz, 1 H), 2.31 (s, 3 H).

Second Step : 7-nitro-2- [4- (N-isopropyl) -nitronyl] phenyl-1,2-benzisoselena

Synthesis of Sol-3 (2H) -one (40)

470 mg of crude product 38 was refluxed with 4 mL of SOCl ₂ for 4 hours. After removal of the remaining thionyl chloride, the crude product 39 was dissolved in CH ₂ Cl ₂ (10 mL). A solution of CH ₂ Cl ₂ (15 mL) containing 100 mg (0.56 mmol) of compound 14 and 0.568 mg (5.61 mmol) of triethylamine was slowly added dropwise to 3 mL of the solution of Compound 39 thus obtained at 0 ° C. After stirring for 2 hours at room temperature, the reaction was concentrated under reduced pressure. The residue was purified by flash column chromatography (silica, EtOAc) to give 121 mg (0.30 mmol) of compound 40 in 53% yield.

¹ H NMR (CDCl ₃ ): δ 8.79 (s, 1H), 8.61 (d, J = 8.07 Hz, 1H), 8.49 (d, J =

7.56 Hz, 1H), 8.03 (d, J = 7.76 Hz, 1H), 7.87 (d, J =

8.10 Hz, 1H), 7.76 (t, J = 7.71 Hz, 1H), 7.55 (s, 2H),

4.28 (septet, J = 6.63 Hz, 1H), 1.56 (d, J = 6.51 Hz,

6H);

¹³ C NMR (CDCl ₃ ): δ 164.03, 142.11, 138.78, 136.52, 135.27, 132.16,

131.42, 131.25, 129.66, 127.95, 127.77, 127.08, 126.41,

124.16, 68.33, 21.05.

The following nitron-containing seleno compounds can be prepared using the methods described in Examples 1-8 above and appropriate starting materials and reagents:

2- [2- (N-isopropyl) nitronyl] -phenyl-1,2-benzisoselenazol-3 (2H) -one;

2- [2- (N-t-butyl) nitronyl] -phenyl-1,2-benzisoselenazol-3 (2H) -one;

5-fluoro-2- [2- (N-isopropyl) nitronyl] -phenyl-1,2-benzisoselenazole-3 (2H) -one;

5-chloro-2- [2- (N-isopropyl) nitronyl] -phenyl-1,2-benzisoselenazol-3 (2H) -one;

5-bromo-2- [2- (N-isopropyl) nitronyl] -phenyl-1,2-benzisoselenazole-3 (2H) -one;

5-methyl-2- [2- (N-isopropyl) nitronyl] -phenyl-1,2-benzisoselenazol-3 (2H) -one;

5-methoxy-2- [2- (N-isopropyl) nitronyl] -phenyl-1,2-benzisoselenazole-3 (2H) -one;

6-chloro-2- [2- (N-isopropyl) nitronyl] -phenyl-1,2-benzisoselenazole-3 (2H) -one;

6-methyl-2- [2- (N-isopropyl) nitronyl] -phenyl-1,2-benzisoselenazole-3 (2H) -one;

5-nitro-2- [2- (N-isopropyl) nitronyl] -phenyl-1,2-benzisoselenazole-3 (2H) -one;

7-nitro-2- [2- (N-isopropyl) nitronyl] -phenyl-1,2-benzisoselenazole-3 (2H) -one;

6,7-methylenedioxy-2- [2- (N-isopropyl) nitronyl] -phenyl-1,2-benzisoselenazole-3 (2H) -one;

2- [3- (N-isopropyl) nitronyl] -phenyl-1,2-benzisoselenazol-3 (2H) -one;

2- [4- (N-isopropyl) nitronyl] -phenyl-1,2-benzisoselenazol-3 (2H) -one;

2- [4- (N-isopropyl) nitronyl] -benzyl-1,2-benzisoselenazol-3 (2H) -one;

2- [4- (N-isopropyl) nitronyl] -phenylethyl-1,2-benzisoselenazol-3 (2H) -one;

2- [4- (N-isopropyl) nitronyl] -pyridin-2-yl-1,2-benzisoselenazol-3 (2H) -one;

2- [5- (N-isopropyl) nitronyl] -pyridin-2-yl-1,2-benzisoselenazol-3 (2H) -one;

2- [4- (N-isopropyl) nitronyl] -pyrimidin-2-yl-1,2-benzisoselenazol-3 (2H) -one;

2- [5- (N-isopropyl) nitronyl] -pyrimidin-2-yl-1,2-benzisoselenazole-3 (2H) -one;

2- [5- (N-isopropyl) nitronyl] -furan-2-yl-1,2-benzisoselenazol-3 (2H) -one;

2- [5- (N-isopropyl) nitronyl] -thiophen-2-yl-1,2-benzisoselenazol-3 (2H) -one;

2- [4- (N-isopropyl) nitronyl] -thiazol-2-yl-1,2-benzisoselenazol-3 (2H) -one;

2- [4- (N-isopropyl) nitronyl] -oxazol-2-yl-1,2-benzisoselenazol-3 (2H) -one;

2- [2- (N-isopropyl) nitronyl] -1H-imidazol-4-yl-1,2-benzisoselenazole-3 (2H) -one;

2- [2- (N-isopropyl) nitronyl] -1-methyl-1H-imidazol-4-yl-1,2-benzisoselenazole-3 (2H) -one;

2- [5- (N-isopropyl) nitronyl] -1H-pyrrol-3-yl-1,2-benzisoselenazole-3 (2H) -one;

2- [5- (N-isopropyl) nitronyl] -1-methyl-1H-pyrrol-3-yl-1,2-benzisoselenazole-3 (2H) -one;

2- [6- (N-isopropyl) nitronyl] -benzothiazol-2-yl-1,2-benzisoselenazol-3 (2H) -one;

2- [5- (N-isopropyl) nitronyl] -2H- [1,2,4] -triazol-3-yl-1,2-benzisoselenazole-3 (2H) -one; And,

2- [5- (N-isopropyl) nitronyl] -2-methyl-2H- [1,2,4] -triazol-3-yl-1,2-benzisoselenazole-3 (2H)- On.

Example 9 Water Solubility Irradiation

Standard solutions were prepared by dissolving accurately measured amounts of test compound (typically 1 mg) in 1 mL of methanol. Beckman DU ^? The UV absorption maximum of each compound was measured with a 7500 spectrophotometer, and the solution was diluted with MeOH as needed.

Subsequently, a small amount of 10 mM phosphate buffer (pH 7.4) was stirred with a magnetic bar for 3 hours in the presence of excess test compound to prepare a saturated solution of each compound. This saturated solution was filtered through a Gelman 0.45 μm filter to remove solid compounds and injected with UV at the wavelength of the maximum absorption measured previously.

Total solubility was determined by the formula: C '= A' (C / A), where C is the concentration of the standard solution (mg / mL); A is the absorbance of the standard solution; A 'is the absorbance of the saturated solution; C 'is the concentration of saturated solution in mg / mL (Protein Sci., 7: 556-563, (1998)). Table 1 summarizes the results.

compound Espelen Example 1 Example 2 Example 5 Example 7 Addition amount (mg) 5.71 5.14 5.55 5.74 5.02 Wavelength (measured value) 330 nm 314 294 302 300 Absorbance (Measured Value) 0.0284 0.6096 0.3584 0.1827 1.2276 Dilution rate One One 10 10 One A ' 0.0284 0.6096 3.584 1.827 1.2276 A 0.6154 1.6807 1.2729 0.8082 0.8871 C (μM) 100 50 50 50 50 C '(μM) = A' (C / A) 4.615 18.135 140.781 113.029 69.192 C '(g / L = mg / mL) 0.001265 0.006516 0.050580 0.041403 0.025830

As shown in Table 1, it can be seen that the compounds of the present invention exhibit much better water solubility than the comparable epselene.

Example 10 Lipid Peroxidation Inhibitory Activity

The antioxidant effect of the compounds of the present invention was examined by inhibiting the radical chain reaction of multilayer liposomes.

Liposomes were prepared as follows: 30 mg of commercially available soybean phosphatidylcholine (PC, Sigma Chemical Co., USA) was dissolved in 1 mL of ethanol, and 200 μl of ethanol / PC solution was stirred with 50 mM NaCl (pH). 7.0 ml) was added to 10 mL of 10 mM Tris buffer.

The ability of the compound to inhibit the oxidation of liposomes was determined as follows: 400 μl of liposome was added to the tube and test compound (dissolved in buffer or ethanol) and histidine-FeCl ₃ (at a final concentration of 167: 33 mM) were added. . FeCl ₂ (at a final concentration of 33 mM prepared in nitrogen-free water) was added to oxidize and the reaction was shaken at 37 ° C. for 15 minutes. Then, a solution of 0.67% TBA (thiobarbituric acid): 10% trichloroacetic acid (2: 1, v / v) was added to a 0.25N HCl solution, and the solution was terminated with t-butylhydroxy. 1 mL of a solution prepared by adding 1.5% (v / v) of toluene (BHT) was added to the tube. The aliquots were heated at 100 ° C. for 20 minutes. After cooling with ice, 1 mL of chloroform was added to 1 mL of the supernatant of the tube, and the tube was centrifuged. The absorbance of the supernatant obtained by centrifugation at 532 nm was measured (see Table 2).

Inhibitor Concentration (IC ₅₀ ) Example 1 Example 2 Example 5 Example 7 S-PBN Epsellen 81.1 μM111.0 μM1.2 μM246.5 μM25.0 μM148.3 μM

As shown in Table 2 above, the compounds of the present invention, in particular the compounds prepared in Example 5, have superior LPO inhibition than the comparative substances S-PBN and eepselen (currently the most anticipated antioxidants in clinical phase III). It can be seen that the activity.

Example 11 Measurement of Glutathione Peroxidase Activity

Glutathione peroxidase-like activity was measured by reduction of GSSG formed through the NADPH-glutathione reductase system as an indicator system.

To 350 μl of 50 mM Tris-HCl (pH 7.6) (assay buffer) containing 5 mM EDTA was added in the following order:

1) 350 μl assay buffer containing 6.4 mM reduced glutathione (GSH), 640 mM nicotinamide adenine dinucleotide (NADPH), and 1.6 unit / mL glutathione disulfide reductase (GR)

2) 70 μl of DMSO in which 800 mM test compound was dissolved (ie each compound was tested at a final concentration of 50 mM).

3) 350 μl of 0.007% t-butyl hydroperoxide prepared by diluting 1 / 10,000-fold with DDW.

The final reaction volume is 1120 μl.

The reaction was advanced at 25 ° C. The glutathione peroxidase activity was examined by measuring the decrease in absorbance at 340 nm for 3 minutes. The electrical activity or initial enzymatic rate is proportional to the slope of the change in absorbance over time.

The oxygen reduction catalytic activity of the tested compounds is consistent with the NADPH consumption rate.

Glutathione peroxidase activity measurement results are shown in Table 3 below. The results are expressed as n-moles of NADPH consumed per minute.

compound Speed A _{340 / min (30 ~ 300sec)} Rate / 0.00622 (nmol NADPH / min / mL) % Axelen Espelen -0.118 18.97 100 Example 1 -0.141 22.67 119.50 Example 2 -0.125 20.10 105.96 Example 5 -0.125 20.13 106.11 Example 7 -0.084 13.50 71.17

As shown in Table 3, the compounds of formula (I) disclosed herein catalyze the reduction of organic hydroperoxides in the presence of glutathione and disulfide glutathione reductase. Thus, it can be seen that the compounds of the present invention have significant and specific glutathione peroxidase activity.

Example 12 Neuronal Protective Action

Example 12-1 : Cerebral cortical neuron culture

Mixed cerebral cortical cell cultures containing neuronal and glial components were prepared from fetal ICR mice at 14 to 15 days of gestation. In brief, dissociated cerebral cortical cells were harvested in 24-multiwell plates (Nunc, And plated on previously prepared glial monolayer cultures at a ratio of 2.5 cerebral hemispheres. Plating medium includes Eagle's minimal essential medium (Earle's salts, glutamine free), 2 mM glutamine, 5% fetal bovine serum (FBS) and 5% serum serum with added glucose (final concentration 20 mM) (horse serum). After 5-6 days of plating, 10 mM cytosine arabinoside was added to the medium to stop the growth of non-neuronal cells. Cultures were placed in a 37 ° C. humid CO ₂ incubator and used for experiments after 10-14 days under laboratory conditions.

Glial cell feeder cultures were prepared from the cerebral neocortex of mice aged 1 to 3 days of age. Dissociated cerebral cortical cells in plating medium supplemented with 5% FBS and 10% horse serum were plated at a rate of 0.25 cerebral hemispheres per 24-multiwell plate. In this way, most neurons do not survive, and only astrocytes survive, resulting in a culture rich in astrocytes. Glial cell cultures were grown for 10-30 days and then used to prepare mixed cerebral cortical cell cultures.

Example 12-2 : Cortical Neuronal Cell Death Protection Induced by Fe ²⁺

When ferrous iron is placed in a solution with normal oxygen pressure, it is self-oxidized to produce active oxygen in the form of hydroxyl radicals, superoxide anion free radicals and hydrogen peroxide. do.

The cerebral cortical cell culture prepared in Example 12-1 was subjected to 30 mM FeCl₂Neuronal cell death was induced by exposure to (Fe) for 24 hours. 37 ° C 5% CO with or without test compound₂Toxin exposure was performed for 24 hours in serum-free Eagle medium (MEM) with 20 mM glucose and 38 mM sodium bicarbonate. All compounds were dissolved in high concentration in DMSO and then diluted to final concentration when added to toxin exposure medium.

Apoptosis measurements were performed in the following manner:

In all experiments, the entire cell damage was assessed by first examining the culture with a phase-contrast microscope. Morphological assessment was performed 1 day after toxin exposure, which is usually the point when most of the cell death processes are completed.

In addition, quantitatively examining total neuronal damage by measuring lactate dehydrogenase (LDH) activity in which damaged or destroyed cells release into extracellular fluid. Small amounts of LDH were always present even in culture media (sham wash controls) that underwent the same exposure procedure except that no toxin was added. This background amount, determined as a sister sham wash control in each experiment, was subtracted from the values obtained from toxin-treated cultures. The absolute value of LDH efflux from toxin exposure was determined by single plate culture. In sister cultures of plating are quite consistent with each other, but in cultures of different plate cultures they are somewhat different. This variability is largely a function of the resulting neuronal density (which varies despite the original plating concentration being constant, perhaps reflecting small differences in cell sample or serum properties). Thus, after 24 hours of 30 mM FeCl ₂ (Fe) exposure in sister cultures, each LDH value was adjusted to the maximum neuronal LDH release (= 100), where almost complete neuronal cell death occurred without glial damage. Levels above 100 generally indicate additional astroglial cell damage.

Figure 1 is a graph showing the results of treatment of both the selenium and Fe ²⁺ toxin.

Figure 2 is a graph showing the result of treating the compound synthesized in Example 1 and Fe ²⁺ toxin together.

Figure 3 is a graph showing the result of treating the compound synthesized in Example 2 and Fe ²⁺ toxin together.

Figure 4 is a graph showing the result of treating the compound synthesized in Example 5 and Fe ²⁺ toxin together.

5 is a graph showing the result of treating the compound synthesized in Example 7 with Fe ²⁺ toxin.

As shown in Figures 1 to 5, the compounds of the present invention was found to effectively protect neuronal cell death by Fe ²⁺ toxin.

Example 13 : Toxicity of Compounds to Neurons

After exposure for 24 hours at different concentrations of test compounds, the viability of the cerebral cortical cells prepared in Example 12-1 was quantified by LDH assay. 24 hour toxin exposure was performed in serum-free Eagle medium (MEM) with 20 mM glucose and 38 mM sodium bicarbonate in 37 ° C. 5% CO ₂ incubator. All compounds were dissolved in high concentration in DMSO and then diluted to final concentration when added to toxin exposure medium.

Cell death was measured in the same manner as in Example 12-2.

Figure 6 is a graph showing the extent of cell damage as the treatment concentration of eepselen increased.

7 is a graph showing the degree of cell damage as the treatment concentration of the compound synthesized in Example 1 increases.

8 is a graph showing the degree of cell damage as the treatment concentration of the compound synthesized in Example 2 increases.

9 is a graph showing the extent of cell damage with increasing treatment concentration of the compound synthesized in Example 5. FIG.

10 is a graph showing the extent of cell damage as the treatment concentration of the compound synthesized in Example 7 increases.

As shown in Figures 6 to 10, it was found that the compounds of the present invention exhibited lower cytotoxicity than that of epselen, suggesting that the compounds can be safely administered in excess.

Example 14 Prevention of Cellular Damage by Ischemia (in vivo)

In this study, male Mongolian gerbils ( Meriones unguiculatus ) weighing 80-88 g were used. Thirty minutes after ischemic injury, rats were orally administered with vehicle, eXelen or various test compounds (dissolved at 60 mg / kg in 10% DMSO). Twenty animals were assigned to each group and mice were general anesthesia with a 2.5% isoflurane mixture in 33% oxygen and 67% nitrous oxide. A midline ventral incision was made at the neck. Both common carotid artery were separated to remove the nerve fibers and closed using a nontraumatic aneurysm clip. Complete blockade of blood flow was confirmed by observing the central artery of the eye with an ophthalmoscope. Five minutes after total carotid artery occlusion, aneurysm clips were removed from both carotid arteries and blood flow recovery (reperfusion) was directly observed under a microscope. The Sham-operated control was subjected to the same surgical procedure except that the carotid artery was not closed. Body temperature was measured and maintained at 37 ° C. ± 0.5 ° C. during surgical treatment and after the period until the animals had fully recovered from anesthesia. At the indicated reperfusion time (4 days), the treated and siamese rats were killed.

Four days after surgery, rats were perfused with PBS (phosphate-buffered saline, pH 7.4) and then 0.1 M phosphate buffer (pH 7.4) in 4% paraformaldehyde. The brain was detached and fixed in fixed fluid such as electricity for 4 hours. Brain tissue was incubated with 30% sucrose overnight to prevent freezing. Subsequently, the samples fixed with the Corny solution were cut into 30 μm sections on cryostat and sequentially stained with Cresyl violet dye.

Staining images of the hippocampus region of each rat were obtained with an Applescanner. Adobe Photoshop version 2.4.1 was used to enhance the brightness and contrast of each image file, and then analyze it with NIH Image 1.59 software. All data obtained from quantified data were analyzed using one-way ANOVA and statistical significance was determined. Bonferroni's test was used for post-hoc comparison. P values below 0.05 or 0.01 were considered statistically significant.

11-a is a graph showing the degree of protection of cell damage in the case of treating the compound of the present invention after ischemia.

11-b is a micrograph showing the degree of protection of cell damage in the case of treating the compound of the present invention after ischemia.

As a result, the test compound prepared in Example 5 shows a better neuroprotective action than iscelene against ischemic neurodegeneration. For the compound synthesized in Example 5, the protective effect was 61% in the group treated with it. The protective effect was 59% in the groups treated with epselen.

In short, we propose that the compound prepared in Example 5 is a potential candidate as a therapeutic agent for diseases associated with ischemia.

As described and demonstrated in detail above, the present invention provides a novel seleno compound containing nitron, a method for preparing the same, and its use for the treatment and / or prevention of various medical diseases caused by free radicals. The compounds of the present invention have a lipid peroxidation (LPO) inhibitory activity that is comparable to or better than that of the reference compounds S-PBN and eepselen. While the compounds are less toxic and more water soluble, they effectively inhibit cerebral neuronal cell death by free radicals and exhibit neuroprotective effects on ischemic neurodegeneration.

From the foregoing description, various modifications and variations of the compositions and methods of the present invention are possible to those skilled in the art. All such modifications within the scope of the appended claims belong to the invention.

Claims (17)

A seleno compound having a nitrone moiety represented by the following formula (I) or a pharmaceutically acceptable salt thereof: (I) Where R ₁ and R ₂ to be equal to or different from each other, hydrogen, halogen, C _{1 ~ 4} - alkyl, C _{1 ~ 4} - Alkoxy, hydroxy, trifluoromethyl, nitro or R ₁ and R ₂ together are methylenedioxy; L is phenyl, C _1-4 alkylphenyl, or furanyl, oxazolyl, isooxazolyl, tee Ophenyl, thiazolyl, isothiazolyl, pyrrolyl, imidazolyl, pyrazolyl, Thiadiazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, ben Unsaturated or saturated heterocyclic click groups having 1 to 4 nitrogen, oxygen or sulfur heteroatoms selected from the group consisting of thiathiazolyl, benzoimidazolyl, benzotriazolyl, triazinyl and triazolyl, wherein hetero Click groups are halogen, C _1-2 alkyl, C _1-4 alkoxy, C _1-4 alkylthio, hydroxy, mercapto, trifluoromethyl, nitro, phenyl, nitrile, carboxy or C _1-4 alkoxycarbonyl Once again by Is substituted twice the same or different); And, R ₃ is isopropyl or t-butyl. The method of claim 1, R ₁ and R ₂ are selected from the group consisting of hydrogen, fluorine, chlorine, bromine, methyl, ethyl, propyl, butyl, hydroxy, methoxy, trifluoromethyl and nitro or R ₁ and R ₂ together are methylene Deoxy; L is phenyl, benzyl, ethylphenyl and furanyl, oxazolyl, thiophenyl, thiazole Reel, pyrrolyl, imidazolyl, pyridyl, pyrimidinyl, benzothiazolyl, Unsaturated or saturated heterocyclic groups having 1 to 4 nitrogen, oxygen or sulfur heteroatoms selected from the group consisting of benzotriazolyl and triazolyl, wherein the heterocyclic groups are fluorine, chlorine, bromine, Once by methyl, ethyl, hydroxy, methoxy, ethoxy, methylsulfanyl, phenylsulfanyl, trifluoromethyl, nitro, phenyl, nitrile, carboxy, methoxycarbonyl or ethoxycarbonyl Two or the same or different from each other); And, R ₃ is selected from the group consisting of isopropyl and t-butyl Seleno compound or pharmaceutically acceptable salt thereof having a nitrone moiety. The method of claim 2, R ₁ and R ₂ are selected from the group consisting of hydrogen, chlorine, bromine, methyl, ethyl, hydroxy, methoxy, trifluoromethyl and nitro or R ₁ and R ₂ together are methylenedioxy; L is phenyl, benzyl, ethylphenyl and furanyl, oxazolyl, thiophenyl, thiazole Reel, pyrrolyl, imidazolyl, pyridyl and pyrimidinyl is selected from the carbonyl groups of 1 to 4 nitrogen, oxygen or sulfur, said unsaturated or saturated heterocyclic ring having from hetero group (wherein the hetero sickle rigs group chlorine , Substituted one or two times with the same or different by methyl, methoxy, methylsulfanyl, phenylsulfanyl, trifluoromethyl, nitro, nitrile, carboxy, methoxycarbonyl or ethoxy carbonyl) Selected from the configured group; And, R ₃ is selected from the group consisting of isopropyl and t-butyl Seleno compound or pharmaceutically acceptable salt thereof having a nitrone moiety. (i) reacting an aldehyde having an appropriate linker (L) and an amino group protected with alkylhydroxylamines (R ₃ NHOH) to obtain nitrons; (ii) deprotecting the compound obtained in step (i) to produce free amine nitron; And, (iii) reacting the free amine of the compound obtained in step (ii) with o-chloroselenobenzoyl chlorides in the presence of an excess base to give the formula (I) Preparing a compound of A process for preparing a compound of formula (I) as defined in claim 1. The method of claim 4, wherein The alkylhydroxylamine of step (i) is characterized by the use of nitroalkane, zinc and acetic acid in a reactor, without separation, from what is produced. A process for preparing a compound of formula (I) as defined in claim 1. The method of claim 4, wherein Step (ii) is characterized by deprotection with trifluoroacetic acid if the protecting group is tert-butoxycarbonyl or LiOH if the protecting group is acetyl. By A process for preparing a compound of formula (I) as defined in claim 1. The method of claim 4, wherein The base of step (iii) is characterized in that the organic base A process for preparing a compound of formula (I) as defined in claim 1. The method of claim 7, wherein The organic base is triethylamine A process for preparing a compound of formula (I) as defined in claim 1. A pharmaceutical composition useful as an antioxidant, comprising an effective amount of a compound of formula (I) as defined in claim 1 as an active ingredient, with one or more pharmaceutically acceptable carriers or excipients. The method of claim 9, The carrier is an oral carrier, characterized in that Pharmaceutical compositions useful as antioxidants. The method of claim 9, The carrier is an injectable carrier, characterized in that Pharmaceutical compositions useful as antioxidants. delete delete delete delete A therapeutic agent for ischemic cerebral nerve disease, comprising the compound of formula (I) as defined in claim 1 as an active ingredient. The method of claim 16, Ischemic cranial nerve disease is characterized by a stroke Ischemic Cerebral Nerve Disease.