KR100454648B1 - N-Alkyl-N-phenylhydroxylamine Compounds Containing Metal Chelating Groups, Their Preparation and Their Therapeutic Uses - Google Patents

Abstract

The present invention provides a novel N -alkyl- N -phenylhydroxylamine derivative containing a metal complex group, a method for preparing the same, and a variety of reactions caused by excess of reactive oxygen species (ROS) and / or zinc (zinc ion). And to their use for the treatment and / or prevention of medical dysfunctions and diseases, in particular stroke, Parkinson's disease and Alzheimer's disease. The compounds of the present invention have a lipid peroxidation (LPO) inhibitory activity that is comparable to or better than that of the comparable compounds, trolox and eepselen. While the compounds are less toxic, they effectively inhibit cerebral neuronal cell death by free radicals (ROS) and / or excess zinc and exhibit neuroprotective effects on ischemic neurodegeneration.

Description

N-Alkyl-N-phenylhydroxylamine Compounds Containing Metal Chelating Groups, Their Preparation and Their Therapeutic Uses {-N-Alkyl-N-phenylhydroxylamine Compounds Containing Metal Complex Groups]

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 neurons 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-Nt-butylnitron (PBN), and various derivatives of PBN are currently being developed. In general, nitron groups increase the solubility of the compounds in water, but they have little inhibitory effect on lipid peroxidation under laboratory conditions ( in vitro ), and they do not provide good protection against brain cells in vivo . (Fevig, Thomas L. et al., J. Med. Chem., 39: 4988-4996 (1996)).

Meanwhile, zinc (Zn), which is present at high concentrations (> 10 mM) in synaptic vesicles, a place for storing neurotransmitters in the brain, is essential for the normal functioning of human cells. Zinc excites neurons and releases them into interstitial fluid at the same time, which plays an important role in neuronal signal transduction, especially by glutamate.

In addition, several studies have reported that zinc present in synapses may play an important role in the pathologic phenomena of the central nervous system. When cortical neurons are exposed to excessive zinc, nerve cells are immediately damaged, at which time the concentration of zinc is similar to the concentration of zinc released from the brain during convulsions or ischemia. Therefore, zinc is presumed to cause nerve cell damage when excessively introduced into neurons. Consistent with the previous hypothesis, selective neuronal damage that occurs after transient global cerebral ischemia has been shown to be due primarily to the translocation of synaptic zinc into neurons rather than to calcium excittotoxicity. (Koh, J.-Y. et al, Science, 272: 1013-1016 (1996); Kim, Y.-H. et al, Neuroscience, 89: 175-182 (1999)). In the case of acute neuronal death from focal ischemia, neurotoxicity due to zinc transfer is more likely to act as a central mechanism. Therefore, it is believed that chelation of zinc will be effective in protecting neurons continuously. However, zinc chelators have not yet been developed for the treatment and prophylaxis of neurodegenerative diseases of the central nervous system.

Summary of the Invention

Therefore, the present inventors introduced a metal complex group to N -alkyl- N -phenylhydroxylamine which will act as a hydrogen atom donor to synthesize a new compound, and zinc, active oxygen while the electric compound shows low toxicity. Having a neuroprotective function against (ROS) and neuro-exciting toxicity factors, it has been confirmed that the present invention can be used as an effective drug in treating and preventing the death of brain cells, thereby completing the present invention.

Finally, a first object of the present invention is to provide a novel N -alkyl- N -phenylhydroxylamine compound containing a metal complex group that is neuroprotective.

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

A third object of the present invention includes an electrical compound as an active ingredient, and is useful for the treatment and / or prevention of medical dysfunctions and diseases such as stroke, Parkinson's disease and Alzheimer's disease caused by excessive oxygen and / or zinc. It is to provide a composition.

A fourth object of the present invention is to provide a method of treating an individual suffering from a disease that requires an antioxidant and / or a metal chelating agent, particularly acute and progressive neurodegenerative diseases, by administering an electropharmaceutical preparation. It is.

The present invention relates to novel N -alkyl- N -phenylhydroxylamine compounds containing metal chelating groups, methods for their preparation and pharmaceutical compositions containing the novel novel compounds as active ingredients. More specifically, the present invention relates to an N -alkyl- N -phenylhydroxylamine compound containing a metal complex group, a method for preparing the same, and an excess of reactive oxygen species (ROS) and / or zinc (zinc ion). It relates to various medical dysfunctions and diseases that are caused, in particular their use for the treatment and / or prevention of stroke, Parkinson's disease and Alzheimer's disease.

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-a is a graph showing the results of treatment with Trolox and Fe ²⁺ toxin (toxin) together.

1-B is a graph showing the results of treatment of Trolox and Zn ²⁺ toxins together.

2-a is a graph showing the result of treating the compound synthesized in Example 1 with Fe ²⁺ toxin.

2-b is a graph showing the results of treatment of the compound synthesized in Example 1 with Zn ²⁺ toxin.

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

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

4-a is a graph showing the results of treating the compound synthesized in Example 3 with Fe ²⁺ toxin.

Figure 4-b is a graph showing the result of treating the compound synthesized in Example 3 and Zn ²⁺ toxin together.

5-a is a graph showing the results of treating the compound synthesized in Example 4 with Fe ²⁺ toxin.

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

6 is a graph showing the extent of cell damage with increasing treatment concentration of trolox.

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 as the treatment concentration of the compound synthesized in Example 3 increases.

10 is a graph showing the extent of cell damage as the treatment concentration of the compound synthesized in Example 4 increases.

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 photomicrograph showing the degree of protection of cell damage in the case of treating the compound of the present invention after ischemia.

In a first embodiment, the present invention provides novel N -alkyl- N -phenylhydroxylamine derivatives containing a metal complex group of the general formula (I):

(I)

Where

R ₁ is branched alkyl, aralkyl, aryl or substituted aryl;

R ₂ is hydrogen or COR ₆ , wherein R ₆ is alkyl or aryl; And,

R ₃ is CO ₂ H, CONH ₂ , or CONR ₄ R _{5 wherein} R ₄ is hydrogen, alkyl or aryl; And, R ₅ is furanyl, oxazolyl, isoxazolyl, thiophenyl, thiazolyl, isothiazolyl, pyrrolyl, imidazolyl, pyrazolyl, oxdiazolyl, thiadiazolyl, pyridyl, pyrimidinyl, Unsaturated or saturated heterocyclic groups having from 1 to 4 nitrogen, oxygen and / or sulfur heteroatoms selected from the group consisting of pyrazinyl, pyridazinyl, triazinyl, triazolyl, tetrazolyl, wherein heterocyclic 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 It may be substituted once or twice the same or different).

Preferred compounds in this context are

R ₁ is branched C _3-8 -alkyl, C _5-8 -cycloalkyl and aryl;

R ₂ is hydrogen, straight or branched C _1-18 alkylcarbonyl, benzoyl, and halogen, C _1-4 -alkyl, C _1-4 -alkoxy, hydroxy, trifluoromethyl, nitro, nitrile, carboxy, alkoxy Benzoyl substituted one or three times with the same or different substituents selected from the group consisting of carbonyl, alkoxycarbonylalkyl and methylenedioxy; And,

R ₃ is CO ₂ H, CONH ₂ or an amide CONR ₄ R ₅ , wherein R ₄ is hydrogen, C _1-4 -alkyl, aralkyl or cycloalkyl; And R ₅ is furanyl, oxazolyl, isoxazolyl, thiophenyl, thiazolyl, isothiazolyl, pyrrolyl, imidazolyl, pyrazolyl, oxadiazolyl, thiadiazolyl, pyridyl, pyrimidinyl, Unsaturated or saturated heterocyclic groups having from 1 to 4 nitrogen, oxygen and / or sulfur heteroatoms selected from the group consisting of pyrazinyl, pyridazinyl, triazinyl, triazolyl, tetrazolyl, wherein heterocyclic Groups are halogen, C _1-2 alkyl, C _1-4 alkoxy, C _1-4 alkylthio, hydroxy, mercapto, trifluoromethyl, nitro, phenyl, nitrile, carboxy and / or C _1-4 alkoxycarbonyl It may be substituted once or twice by the same or different).

More preferred compound,

R ₁ is isopropyl, t-butyl, cyclopentyl, cyclohexyl, phenyl;

R ₂ is hydrogen, acetyl, propionyl, benzoyl; And,

R ₃ is CO ₂ H, CONH ₂ or an amide CONR ₄ R _{5 wherein} R ₄ is hydrogen, methyl, ethyl, propyl, benzyl, cyclopentyl or cyclohexyl; And R ₅ consists of oxazolyl, thiazolyl, isothiazolyl, imidazolyl, pyrazolyl, thiadiazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, triazinyl, triazolyl, tetrazolyl Unsaturated or saturated heterocyclic groups having from 1 to 4 nitrogen, oxygen and / or sulfur heteroatoms selected from the group, wherein the heterocyclic group is fluorine, chlorine, bromine, methyl, methoxy, ethoxy, methyl Mercapto, trifluoromethyl, nitro, phenyl, nitrile, carboxy, methoxycarbonyl and / or ethoxycarbonyl, which may be substituted one or two times the same or different).

The compounds of the present invention show similar or better LPO inhibitory activity compared to the comparable compounds Trolox and Epsellene. The compounds of the present invention also exhibit lower toxicity than these, but effectively inhibit brain cell death induced by free radicals (ROS) and / or zinc, and have a neuroprotective effect on ischemic neuronal degeneration. Indicates.

The compounds of the present invention, in particular the compounds synthesized in Example 1 below, have very low toxicity of LD ₅₀ ≧ 7,000 mg / kg orally when administered in rats and ≧ 750 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 espelen (LD ₅₀ values of espelen obtained from mice are determined by oral administration). More than 6,810 mg / kg and more than 740 mg / kg when administered intraperitoneally Similarly, the LD ₅₀ value of the epsilen obtained from rats was more than 6,810 mg / kg when administered orally, and when administered intraperitoneally More than 580 mg / kg). Thus, administration of large amounts of the novel compounds immediately following a stroke or other trauma can significantly reduce oxidative damage in many cases.

In a second embodiment, the present invention provides a process for the preparation of a compound of formula (I) wherein R ₃ is CONR ₄ R ₅ as shown in the following reaction pathway:

The O-protected alkylhydroxyaminobenzaldehyde, 1, is oxidized to give carboxylic acid, 2 , which is then amide-linked with HNR ₄ R ₅ to give amide, 3 . After removal of the protecting group, the N-hydroxy group of compound 4 is converted to the N-acylhydroxy group by acylating in the presence of a suitable base used for pharmaceutical use.

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. Preferably, there is provided a neuroprotective agent comprising the compound of formula (I) as an active ingredient.

In a fourth embodiment, the present invention comprises administering an electropharmaceutical composition to a subject suffering from a disease, particularly acute and progressive neurodegenerative disorders, which require antioxidants and / or metal complexing agents. Provide a way to.

As mentioned above, the compounds of the present invention have been found to effectively mitigate various effects due to free radicals and / or zinc. 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 invention are generally administered in the form of a pharmaceutical composition comprising at least one active compound of the invention and comprising a pharmaceutically acceptable carrier or carrier suitable for use in pharmaceutical 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 N -alkyl- N -phenylhydroxylamine compounds containing the metal complex groups of the invention are usually minor components (other residues for treating acids that are useful as carriers or carriers and useful for making the desired dosage form). About 0.1 to 50% or preferably about 1 to 40% of the total weight including water).

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 described 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 4- [N- (t-butyl) -N-hydroxyamino] -N-thiazol-2-yl-benzami

Synthesis of De (4)

First step : 4- [N- (t-butyl) -N- (t-butyldimethylsilyloxy) amino] -benzoic acid (2)

synthesis

Slowly add acetone (10 mL) in which 2,2 g (7.15 mmol) of aldehyde 1 is dissolved in a solution of H ₂ O / acetone (21: 3, v / v, 24 mL) containing 1.922 g (12.16 mmol) KMnO ₄ . After that, the mixture was stirred at room temperature for 10 hours. After completion of the reaction, the mixture was filtered through a Celite pad and acetone was removed under reduced pressure. The aqueous phase was acidified with 1N HCl and extracted with CH ₂ Cl ₂ , dried over anhydrous MgSO ₄ , filtered and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica, Hex: EtOAc = 3: 1) to afford 1.97 g of compound 2 in 85% yield.

¹ H NMR (CDCl ₃ ): δ 7.73 (d, 2H, J = 8.8 Hz), 7.40 (d, 2H, J = 8.2 Hz),

1.12 (s, 9H) 0.92 (s, 9H), -0.13 (br s, 6H)

¹³ C NMR (CDCl ₃ ): δ 172.1, 157.0, 129.7, 124.8, 61.9, 26.4, 26.2 18.0,

-4.5

Second Step : 4- [N- (t-butyl) -N- (t-butyldimethylsilyloxy) amino] -N-thiazol-2-yl-

Synthesis of Benzamide (3)

2.2 g (11.2 mmol) of 1- (3-dimethylamino) propyl-3-ethylcarbodiimide hydrochloride (EDC), 0.566 g (0.463) in DMF (5 mL) solution of 3 g (9.27 mmol) dissolved in compound 2 mmol) 4-dimethylaminopyridine (DMAP) and 1.11 g (13.9 mmol) 2-aminothiazole were added and stirred at room temperature for 24 hours before H ₂ O and CH ₂ Cl ₂ were added. The organic layer was separated, dried over anhydrous MgSO ₄ , filtered and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica, Hex: EtOAc = 5: 1) to afford 3.26 g of compound 3 in 87% yield.

¹ H NMR (CDCl ₃ ): δ 7.93 (d, 2H, J = 8.7 Hz), 7.38 (d, 2H, J = 8.1 Hz),

6.98 (d, 1H, J = 3.7 Hz), 6.88 (d, 1H, J = 3.6 Hz), 1.14

(s, 9H), 0.94 (s, 9H), -0.06 (br s, 6H)

¹³ C NMR (CDCl ₃ ): δ 166.1, 160.9, 155.9, 137.1, 129.1, 127.8,

125.1, 113.2, 61.6, 26.3, 26.2, 18.0, -4.5

Third Step : 4- [N- (t-butyl) -N-hydroxyamino] -N-thiazol-2-yl-benzamide

Synthesis of 4

40 to 50% solution of 40% trifluoroacetic acid (TFA: H ₂ O: CH ₂ Cl ₂ = 1: 0.5: 1, v / v) in which 2.5 g (6.16 mmol) of Compound 3 was dissolved The reaction was stirred at 占 폚 for 9 hours, and after the reaction was completed, 10 mL of water was added to stop the reaction. After adding ethyl acetate and saturated NaHCO ₃ solution to the reaction mixture, the organic layer was separated, dried over anhydrous MgSO ₄ , filtered and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica, Hex: EtOAc = 2: 1) to yield 1.5 g of compound 4 in 84% yield.

¹ H NMR (DMSO-d ₆ ): δ 12.45 (s, NH), 8.51 (s, OH), 8.01 (d, 2H, J = 8.7

Hz), 7.54 (d, 1H, J = 3.5 Hz), 7.32 (d, 2H, J = 8.7 Hz),

7.25 (d, 1H, J = 3.6 Hz), 1.12 (s, 9H)

¹³ C NMR (DMSO-d ₆ ): δ 164.6, 158.8, 154.9, 137.5, 127.7, 126.9, 123.2,

113.63, 60.1, 26.2

Example 2 : 4- [N- (t-butyl) -N-hydroxyamino] -N-pyridin-2-yl-benzami

Synthesis of de (6)

First Step : 4- [N- (t-butyl) -N- (t-butyldimethylsilyloxy) amino] -N-pyridine-2-

Synthesis of yl-benzamide (5)

0.189 g of 0.733 g (3.71 mmol) 1- (3-dimethylamino) propyl-3-ethylcarbodiimide hydrochloride in a solution of CH ₂ Cl ₂ (5 mL) in which 1.0 g (3.09 mmol) of Compound 2 was dissolved 1.545 mmol) of 4-dimethylaminopyridine and 0.438 g (4.63 mmol) of 2-aminopyridine were added and stirred at room temperature for 12 hours before H ₂ O and ethyl acetate were added. The organic layer was separated, washed with saturated NaHCO ₃ solution, dried over anhydrous MgSO ₄ , filtered and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica, Hex: EtOAc = 10: 1) to give 2.3 g of compound 5 in 75% yield.

¹ H NMR (CDCl ₃ ): δ 8.58 (s, OH), 8.38 (d, 1H, J = 8.3 Hz), 8.28 (s, 1H),

7.8 (d, 2H, J = 8.0 Hz), 7.75 (t, 1H, J = 8.4 Hz), 7.36

(d, 2H, J = 8.3 Hz), 7.05 (t, 1H, J = 5.5 Hz), 1.12 (s,

9H), 0.92 (s, 9H), -0.13 (br s, 6H)

¹³ C NMR (CDCl ₃ ): δ 166.1, 155.3, 152.1, 147.8, 138.5, 130.6, 126.9,

125.1, 119.6, 114.5, 61.5, 26.3, 26.2, 18.0, -4.5

Second Step : 4- [N- (t-butyl) -N-hydroxy-amino] -N-pyridin-2-yl-benza

Synthesis of Mead 6

2.84 mL (2.84 mmol) of THF solution containing tetrabutylammonium fluoride at a concentration of 1 M was added to a THF (3 mL) solution in which Compound 5 was dissolved 0.95 g (2.37 mmol) at room temperature and stirred for 1 hour. . After the reaction was complete, 2 mL of water was added to stop the reaction. After adding ethyl acetate and saturated NaHCO ₃ solution to the reaction mixture, the organic layer was separated, dried over anhydrous MgSO ₄ , filtered and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica, Hex: EtOAc = 10: 1) to afford 0.44 g of compound 6 in 65% yield.

¹ H NMR (DMSO-d ₆ ): δ 10.58 (s, 1H), 8.47 (s, OH), 8.40 (s, 1H), 8.17 (d,

1H, J = 8.4 Hz), 7.95 (d, 2H, J = 8.4 Hz), 7.82 (t, 1H,

J = 7.1 Hz), 7.29 (d, 2H, J = 8.2 Hz), 7.15 (t, 1H, J =

5.7 Hz), 1.12 (s, 9H)

¹³ C NMR (DMSO-d ₆ ): δ 164.7, 153.6, 151.5, 147.5, 137.6, 128.3, 126.9,

122.6, 119.1, 114.1, 59.2, 26.3

Example 3 : 4- [N- (t-butyl) -N-hydroxyamino] -N-pyridin-2-yl-methyl-bene

Synthesis of Zamide (8)

First Step : 4- [N- (t-butyl) -N- (t-butyldimethylsilyloxy) amino] -N-pyridine-2-

Synthesis of mono-methyl-benzamide (7)

0.11 g (0.561 mmol) of 1- (3-dimethylamino) propyl-3-ethylcarbodiimide hydrochloride, 0.03 g (1) in 0.15 g (0.464 mmol) dissolved CH ₂ Cl ₂ (10 mL) solution 0.232 mmol) 4-dimethylaminopyridine and 0.072 mL (0.695 mmol) 2-aminomethylpyridine were added and stirred at room temperature for 2 hours, followed by H ₂ O and ethyl acetate. The organic layer was separated, dried over anhydrous MgSO ₄ , filtered and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica, Hex: EtOAc = 1: 1) to yield 0.13 g of compound 7 in 70% yield.

¹ H NMR (CDCl ₃ ): δ 8.55 (d, 1H, J = 0.7 Hz), 7.75 (d, 2H, J = 8.9 Hz),

7.67 (t, 1H, J = 8.5 Hz), 7.58 (s, 1H), 7.32 (d, 2H, J =

8.4 Hz), 7.27 (s, 1 H), 7.19 (t, 1 H, J = 6.0 Hz), 4.74 (d,

2H, J = 4.8 Hz), 1.08 (s, 9H), 0.89 (s, 9H), -0.14 (br s,

6H)

Second Step : 4- [N- (t-butyl) -N-hydroxyamino] -N-pyridin-2-yl-methyl-benz

Synthesis of Amide 8

0.73 mL (0.73 mmol) of THF solution containing tetrabutylammonium fluoride at a concentration of 1 M was added to a THF (1 mL) solution containing 0.06 g (0.145 mmol) of Compound 7 at room temperature, followed by stirring for 24 hours. . After the reaction was complete, 2 mL of water was added to stop the reaction. After adding ethyl acetate and saturated NH ₄ Cl solution to the reaction mixture, the organic layer was separated, dried over anhydrous MgSO ₄ , filtered and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica, Hex: EtOAc = 1: 2) to afford 0.026 g of compound 8 in 60% yield.

¹ H NMR (DMSO-d ₆ ): δ 8.50 (d, 1H, J = 3.9 Hz), 8.44 (s, 1H), 7.81 (d, 2H,

J = 8.3 Hz), 7.74 (t, 1H, J = 7.2 Hz), 4.52 (d, 2H, J =

5.5 Hz), 1.18 (s, 9H)

¹³ C NMR (DMSO-d ₆ ): δ 166.0, 158.9, 153.7, 148.7, 136.6, 129.4, 126.6,

123.3, 121.9, 120.8, 59.8, 44.6, 26.1,

Example 4 : 4- [N- (t-butyl) -N-hydroxyamino] -N- (8-hydroxyquinoline-5-

Synthesis of yl) -benzamide (10)

First step : 4- [N- (t-butyl) -N- (t-butyldimethylsilyloxy) amino] -N- (8-hydroxy

Synthesis of Cyquinolin-5-yl) -benzamide (9)

Compound 2 0.15g (0.464mmol) 1- (3- dimethylamino) is of 0.11g (0.561 mmol) in CH ₂ Cl ₂ (2mL) solution of dissolved propyl-3-ethylcarbodiimide hydrochloride, 0.03g ( 0.232 mmol) of 4-dimethylaminopyridine, 2 mL of triethylamine and 0.16 g (0.695 mmol) of 5-amino-8-hydroxyquinoline HCl salt were added and stirred for 24 hours, followed by H ₂ O and ethyl acetate The organic layer was separated, dried over anhydrous MgSO ₄ , filtered and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica, Hex: EtOAc = 1: 1) to afford 0.076 g of compound 9 in 35% yield.

¹ H NMR (CDCl ₃ ): δ 8.86 (d, 1H, J = 2.7 Hz), 8.20 (d, 2H, J = 8.9 Hz),

8.16 (d, 1H, J = 5 Hz), 7.35-7.32 (m, 2H), 6.75 (d, 1H,

J = 8.1 Hz)

Second Step : 4- [N- (t-butyl) -N-hydroxyamino] -N- (8-hydroxyquinoline-5-

Synthesis of yl) -benzamide (10)

0.43 mL (0.43 mmol) of THF solution in which tetrabutylammonium fluoride was dissolved at a concentration of 1 M was added to THF (1 mL) solution in which 0.09 g (0.085 mmol) of Compound 9 was dissolved, and stirred for 1 hour. After the reaction was complete, 2 mL of water was added to stop the reaction. After adding ethyl acetate and saturated NH ₄ Cl solution to the reaction mixture, the organic layer was separated, dried over anhydrous MgSO ₄ , filtered and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica, Hex: EtOAc = 10: 1) to afford 0.021 g of compound 10 in 70% yield.

¹ H NMR (DMSO-d ₆ ): δ 8.73 (d, 1H, J = 2.9 Hz), 8.60 (s, 1H), 8.56 (s,

1H), 8.04 (d, 2H, J = 8.3 Hz), 7.44 (s, 1H), 7.40 (d,

2H, J = 8.0 Hz), 7.32 (d, 1H, J = 8.1 Hz), 6.72 (d, 1H,

J = 8.1 Hz), 5.99 (s, OH), 1.16 (s, 9H)

N -alkyl- N -phenylhydroxylamine compounds containing the following metal complex groups can be prepared using the methods disclosed in Examples 1-4 above and appropriate starting materials and reagents:

2- ( N -t-butyl- N -hydroxyamino) -N- furan-2-yl-benzamide

3- ( N -t-butyl- N -hydroxyamino) -N- furan-2-yl-benzamide

4- ( N -t-butyl- N -hydroxyamino) -N- furan-2-yl-benzamide

4- ( N -cyclohexyl- N -hydroxyamino) -N- furan-2-yl-benzamide

4- ( N -t-butyl- N- acetoxyamino) -N- furan-2-yl-benzamide

4- ( N - cyclohexyl- N- acetoxyamino) -N- furan-2-yl-benzamide

4- ( N -t-butyl- N- benzoyloxyamino) -N- furan-2-yl-benzamide

4- ( N -cyclohexyl- N- benzoyloxyamino) -N- furan-2-yl-benzamide

4- ( N -t-butyl- N -hydroxyamino) -N -oxazol-2-yl-benzamide

4- ( N -cyclohexyl- N -hydroxyamino) -N -oxazol-2-yl-benzamide

4- ( N -t-butyl- N -hydroxyamino) -N -thiophen-2-yl-benzamide

4- ( N -cyclohexyl- N -hydroxyamino) -N -thiophen-2-yl-benzamide

2- ( N -t-butyl- N -hydroxyamino) -N -thiazol-2-yl-benzamide

3- ( N -t-butyl- N -hydroxyamino) -N -thiazol-2-yl-benzamide

4- ( N -t-butyl- N -hydroxyamino) -N -thiazol-2-yl-benzamide

4- ( N -cyclohexyl- N -hydroxyamino) -N -thiazol-2-yl-benzamide

4- ( N -t-butyl- N- acetoxyamino) -N -thiazol-2-yl-benzamide

4- ( N - cyclohexyl- N- acetoxyamino) -N -thiazol-2-yl-benzamide

4- ( N -t-butyl- N- benzoyloxyamino) -N -thiazol-2-yl-benzamide

4- ( N -cyclohexyl- N- benzoyloxyamino) -N -thiazol-2-yl-benzamide

4- (N- t- butyl-N- hydroxy-amino) - N- (1H- imidazol-2-yl-benzamide

4- (N- Cyclo hexyl-N- hydroxy-amino) - N- (1H- imidazol-2-yl-benzamide

4- (N- t- butyl-N- hydroxy-amino) - N- (1- methyl -1H- imidazol-2-yl-benzamide

4- (N- Cyclo hexyl-N- hydroxy-amino) - N- (1- methyl -1H- imidazol-2-yl-benzamide

4- (N- t- butyl-N- hydroxy-amino) - N- (2H- pyrazol-3-yl) -benzamide

4- (N- Cyclo hexyl-N- hydroxy-amino) - N- (2H- pyrazol-3-yl) -benzamide

4- (N- t- butyl-N- hydroxy-amino) - N- (2- methyl -2H- pyrazol-3-yl) -benzamide

4- (N- Cyclo hexyl-N- hydroxy-amino) - N- (2- methyl -2H- pyrazol-3-yl) -benzamide

2- ( N -t-butyl- N -hydroxyamino) -N- pyridin-2-yl-benzamide

3- ( N -t-butyl- N -hydroxyamino) -N- pyridin-2-yl-benzamide

4- ( N -t-butyl- N -hydroxyamino) -N- pyridin-2-yl-benzamide

4- ( N -cyclohexyl- N -hydroxyamino) -N- pyridin-2-yl-benzamide

4- ( N -t-butyl- N- acetoxyamino) -N- pyridin-2-yl-benzamide

4- ( N - cyclohexyl- N- acetoxyamino) -N- pyridin-2-yl-benzamide

4- ( N -t-butyl- N- benzoyloxyamino) -N- pyridin-2-yl-benzamide

4- ( N -cyclohexyl- N- benzoyloxyamino) -N- pyridin-2-yl-benzamide

4- ( N -t-butyl- N -hydroxyamino) -N- pyrimidin-2-yl-benzamide

4- ( N -cyclohexyl- N -hydroxyamino) -N- pyrimidin-2-yl-benzamide

4- ( N -t-butyl- N -hydroxyamino) -N -pyridazin-3-yl-benzamide

4- ( N -cyclohexyl- N -hydroxyamino) -N -pyridazin-3-yl-benzamide

4- ( N -t-butyl- N -hydroxyamino) -N- pyrazin-2-yl-benzamide

4- ( N -cyclohexyl- N -hydroxyamino) -N- pyrazin-2-yl-benzamide

4- (N- t- butyl-N- hydroxy-amino) - N- [1,3,5] triazin-2-yl-benzamide

4- (N- Cyclo hexyl-N- hydroxy-amino) - N- [1,3,5] triazin-2-yl-benzamide

4- (N- t- butyl-N- hydroxy-amino) - N- (2H- [1,2,4] triazol-3-yl) -benzamide

4- (N- Cyclo hexyl-N- hydroxy-amino) - N- (2H- [1,2,4] triazol-3-yl) -benzamide

4- (N- t- butyl-N- hydroxy-amino) - N- (2- methyl -2H- [1,2,4] triazol-3-yl) -benzamide

4- (N- Cyclo hexyl-N- hydroxy-amino) - N- (2- methyl -2H- [1,2,4] triazol-3-yl) -benzamide

4- ( N -t-butyl- N -hydroxyamino) -N- (1H-tetrazol-5-yl) -benzamide

4- (N- Cyclo hexyl-N- hydroxy-amino) - N- (1H- tetrazol-5-yl) -benzamide

4- ( N -t-butyl- N -hydroxyamino) -N- (1-methyl-1H-tetrazol-5-yl) -benzamide

4- (N- Cyclo hexyl-N- hydroxy-amino) - N- (1- methyl -1H- tetrazol-5-yl) -benzamide

Example 5: 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 prepared by adding 0.67% TBA (thiobarbituric acid): 10% trichloroacetic acid (2: 1, v / v) to 0.25N HCl solution was prepared, and the solution was terminated with t-butylhydride. 1 mL of a solution prepared by adding 1.5% (v / v) of oxytoluene (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 1).

Inhibitor concentration (IC₅₀) Example 1 1.44 μM Example 2 4.30 μM Example 3 11.05 μM Example 4 61.85 μM Epsellene 68.86 μM Trolox 10.25 μM

As shown in Table 1, the compounds of the present invention is a comparative material Trolox (vitamin E derivatives and various laboratory conditions (in vitro) And in vivo conditions (in vivo) Is used as a comparative substance in antioxidant experiments) and Epsellene (Clinical as the most anticipated antioxidant currently Better than in stage III) LPO It can be seen that the inhibitory activity.

Example 6 Neuronal Protective Activity by the Present Compound

Example 6-1 : Cerebral Cortical Neuron Culture

Mixed cerebral cortical cell cultures containing neuronal and glial components were prepared from fetal ICR mice at 14-15 days of gestation. Briefly, dissociated cerebral cortical cells were plated onto previously prepared glial monolayer cultures at a rate of 2.5 cerebral hemispheres per 24-multiwell plate (Nunc, USA). Plating medium is Eagle's minimal essential medium (Earle's salts, without glutamine) added with glucose (final concentration 20 mM), 2 mM glutamine, 5% fetal bovine serum (FBS) and 5% It consists of 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 6-2 : Cortical Neuronal Cell Death Protection Induced by Fe ²⁺ and Zn ²⁺

Action

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.

Cortical cell cultures prepared in Example 6-1 were exposed to 30 μM FeCl ₂ (Fe) or 35 μM ZnCl ₂ (Zn, neuro-excitatory factor) for 24 hours to induce neuronal cell death. Toxin exposure was performed for 24 hours in serum-free Eagle medium (MEM) with 20 mM glucose and 38 mM sodium bicarbonate in 37 ° C. 5% CO ₂ incubator with or without test compound. 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. A small amount of LDH was always present even in culture medium (sham wash control) 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 the toxin treated cultures. Absolute values of LDH effluent resulting from toxin exposure are quite consistent with each other in sister cultures of single plating, but somewhat different in cultures of different plates. 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) or 35 mM ZnCl ₂ (Zn) exposure in sister cultures, each LDH value was adjusted to the maximum neuronal LDH release (= 100), where nearly complete neuronal cell death without glial cell damage. This happens. Levels above 100 generally indicate additional astroglial cell damage.

1-a is a graph showing the results of treatment of trolox and Fe ²⁺ toxin together.

1-b is a graph showing the results of treating Trolox and Zn ²⁺ toxins together.

2-a is a graph showing the result of treating the compound synthesized in Example 1 with the Fe ²⁺ toxin.

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

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

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

4-a is a graph showing the results of treating the compound synthesized in Example 3 with the Fe ²⁺ toxin.

4-b is a graph showing the result of treating Zn ²⁺ toxin with the compound synthesized in Example 3. FIG.

5-a is a graph showing the results of treating the compound synthesized in Example 4 with the Fe ²⁺ toxin.

5-b is a graph showing the result of treating the compound synthesized in Example 4 with Zn ²⁺ toxin together.

As can be seen in Figures 1-a to 5-b, it was found that the compounds of the present invention effectively protect against cell death by Fe ²⁺ or Zn ²⁺ toxins, which are present in excess of free radicals and / or zinc. This suggests that it can be used as a neuroprotective agent to protect brain nerve cells from (neuro-excitatory toxicity factor).

Example 7 Neuronal Toxicity of Compounds

After exposure for 24 hours at different concentrations of test compounds, the viability of the cerebral cortical cells prepared in Example 6-1 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 6-2.

6 is a graph showing the extent of cell damage with increasing treatment concentration of trolox.

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 as the treatment concentration of the compound synthesized in Example 3 increases.

10 is a graph showing the extent of cell damage as the treatment concentration of the compound synthesized in Example 4 increases.

As shown in Figures 6 to 10, the compounds of the present invention exhibit low cytotoxicity, suggesting that the compounds can be safely administered in excess.

Example 8 Prevention of Cell 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 same surgical procedures were performed in the sham-operated control, 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 the quantified data were analyzed using one-way ANOVA and statistical significance was determined. Bonferroni's test was used for post-hoc comparison. P values less than 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 1 exhibits superior neuroprotective action than iscelene against ischemic neurodegeneration. For the compound synthesized in Example 1, the protective effect was 81% 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 1 is a possible substance as a therapeutic for ischemia.

As described and demonstrated in detail above, the present invention provides a novel N -alkyl- N -phenylhydroxylamine compound containing a metal complex group, a preparation method thereof, and various medical diseases caused by free radicals and / or excess zinc. Its use is provided for the treatment and / or prevention of. The compounds of the present invention have a lipid peroxidation (LPO) inhibitory activity that is comparable to or better than that of the comparable compounds, trolox and eepselen. While the compounds are less toxic, they effectively inhibit cerebral neuronal cell death by free radicals and / or excess zinc 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 (12)

N -alkyl- N -phenylhydroxylamine derivatives containing a metal complex group represented by the following formula (I) and pharmaceutically acceptable salts thereof: (I) Where R ₁ is branched C _3-8 -alkyl, C _5-8 -cycloalkyl and phenyl; R ₂ is hydrogen, straight or branched C _1-18 alkylcarbonyl, benzoyl, and halogen, C _1-4 -alkyl, C _1-4 -alkoxy, hydroxy, trifluoromethyl, nitro, nitrile, carboxy, Benzoyl substituted one or three times with the same or different substituents selected from the group consisting of alkoxycarbonyl, alkoxycarbonylalkyl and methylene dioxy; And, R ₃ is CO ₂ H, CONH ₂ or amide CONR ₄ R ₅ , wherein R ₄ is hydrogen, C _1-4 -alkyl, benzyl or cycloalkyl; And R ₅ is furanyl, oxazolyl, iso-ok Sazolyl, thiophenyl, thiazolyl, isothiazolyl, pyrrolyl, imidazole Unsaturated or saturated heterocyclic click groups selected from the group consisting of reyl, pyrazolyl, oxadizolyl, thiadiazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, triazinyl, triazolyl, tetrazolyl, wherein , A 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 is substituted once or twice by the same or different). delete The method of claim 1, R ₁ is isopropyl, t-butyl, cyclopentyl, cyclohexyl, phenyl; R ₂ is hydrogen, acetyl, propionyl, benzoyl; And, R ₃ is CO ₂ H, CONH ₂ or an amide CONR ₄ R _{5 wherein} R ₄ is hydrogen, methyl, ethyl, propyl, benzyl, cyclopentyl or cyclohexyl; And R ₅ is oxazolyl, thiazolyl, isothiazolyl, imidazolyl, pyrazolyl, thiadiazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, triazinyl, triazolyl, tetrazolyl Unsaturated or saturated heterocyclic groups selected from the group consisting of fluorine, chlorine, bromine, methyl, methoxy, ethoxy, methyl mercapto, hydroxy, mercapto, trifluor Romethyl, nitro, phenyl, nitrile, carboxy, methoxycarbonyl or ethoxycarbonyl, one or two times being substituted by the same or different); N -alkyl- N -phenylhydroxylamine derivatives containing metal complex groups and pharmaceutically acceptable salts thereof. (i) oxidizing O-protected alkylhydroxyaminobenzaldehyde ( 1 ) to obtain carboxylic acid ( 2 ); (ii) amide-linking the compound obtained in step (i) with HNR ₄ R ₅ to obtain an amide ( 3 ); (iii) deprotecting the reaction of the compound obtained in step (ii); And, (iv) O-acylating the N-hydroxy group of Compound ( 4 ) to produce an N-acylhydroxy group A process for preparing a compound of formula (I) as defined in claim 1. A therapeutic agent for acute or progressive cerebral neurodegenerative disease, comprising an effective amount of a compound of formula (I) as defined in claim 1 as an active ingredient, together with one or more pharmaceutically acceptable carriers or excipients. A neuroprotective agent comprising an effective amount of a compound of formula (I) as defined in claim 1 as an active ingredient, together with one or more pharmaceutically acceptable carriers or excipients. The method of claim 6, The carrier is characterized in that the carrier for oral administration Neuroprotective agents. The method of claim 6, The carrier is characterized in that the carrier for injection Neuroprotective agents. delete A method of treating acute or progressive neurodegenerative disorders comprising administering a compound of formula (I) as defined in claim 1 to a mammal, excluding a human, in an amount effective to alleviate the disease. The method of claim 10, Acute or progressive neurodegenerative disorders are selected from the group consisting of stroke, Parkinson's disease and Alzheimer's disease. A method of treating acute or progressive neurodegenerative disorders. The method of claim 10, Mammals other than humans are characterized by a stroke A method of treating acute or progressive neurodegenerative disorders.